Vican 22 12 2020 1

Virtual International Conference
on
01st November 2020
Jointly Organized
by
Association of Global Academicians and

Researchers (AGAR)
&
Association of Indian Biologists (AIB)
Tamil Nadu, India
ISBN: 978-81-948129-1-3
Virtual International Conference on Advancements in
Nanotechnology [VICAN-2020]
Jointly Organized by
Association of Global Academicians and Researchers (AGAR)
and
Association of Indian Biologists (AIB), Tamil Nadu, INDIA
Date: 01-11-2020
Keynote Speakers
Dr. Rajeshkumar
Dr. Periyathambi Dhaiveegan. Department of Pharmacology
Technical Department, Saveetha Dental College and Hospitals
Tripod Nano Technology Corporation, Saveetha Institute of Medical and
Taoyuan, Taiwan. Technical Sciences (SIMATS),
Title: Colloidal Metal Nanoparticles. Saveetha University,
Chennai, India
Title: Nanoparticles in Recent
biomedical applications.
Prof. Yusuf Sarkingobir Dr. Pradeep Kumar Panda
Department of Biology, Department of Chemical and Materials
Shehu Shagari College of Education, Engineering,
Sokoto, Sokoto State, Nigeria. Chang Gung University,
Title: Harmful Biological Effects of Taoyuan, Taiwan.
Plastics. Title: Introduction to Material science.
MESSAGE FROM EDITOR-I
It is indeed a pleasure and honour to be part and parcel of Virtual International Conference
on Advancements in Nanotechnology jointly organised by Association of Indian Biologists (AIB),
and Association of Global Academicians and Researchers (AGAR)
(AGAR), Tamil Nadu,
Nadu which has been
organised with a great magnitude in the era of COVID-19. This remains as a history due to its
tremendous response across the globe. I am indeed grateful to the members of the association
assoc for
providing me an opportunity and for reposing faith in me. All this has been made possible with
their guidance. My thanks to the faculty members and students who have participated in this
conference. I am very thankful to other Editors and Associate Editors for guiding me in times of
need. I am very fortunate and blessed to be part of this prestigious conference.
With Regards,
Dr. SHEKHAR R,
Associate Professor and Head,
Department of Computer Science and Engineering,
Alliance University,
Bangalore,
Karnataka.
Email: shekhar.aims@gmail.com
MESSAGE FROM EDITOR
EDITOR-II
Dear Friends,
It is wonderful to see the Association of Indian Biologists (AIB), Tamil Nadu taking up an
important experimental education and research strategies and at the same time an important
problem in the society- Virtual International Conference on Advancements in Nanotechnology
more and younger generations.
The topic ‘education and research’ gives mu
much
ch room to search for the latest trends in
dealing with important education role and emerging research strategies. The awareness gained
during this seminar would reach the student community so that the education we receive becomes
directly useful to the society.
We need a more strategic and holistic education and research approach to integrate aspects
from the different field of research. A seminar of this type will broaden the minds of the young
researchers to search for new solutions to real life strateg
strategies.
I wish all success to the organizing committee of this seminar and associations for hosting
this valuable academic exercise. Congratulations and God Bless Your Effort.
With Regards,
Dr. JAGANNATHAN S, S
Assistant Research Officer, Officer In Charge,
TCARV &QCD Stability Departments,
Pasteur Institute of India,
Coonoor-643103,
The Nilgiris, Tamilnadu,
Email: seljag2005@gmail.com
MESSAGE FROM EDITOR
EDITOR-III
Dear All,
Indeed it is a privilege and honour to be part and parcel of the jointly organised
Virtual International Conference on Advances in Nanotechnology by the Association of
Indian Biologists (AIB), Tamil Nadu, organised with great magnitude in the COVID-19
period. Because of its immense reaction around the globe, this remains a fact. I am indeed
thankful to the members of the association for providing me with an opportunity and for
restoring trust in me with their support, all this was made possible
possible.. My gratitude to the
representatives of the faculty and the students who participated in this meeting. I'm so
very grateful to my associates for having driven me in times of need. I am blessed and
very lucky to be part of this prestigious meeting.
Thanks to the keynote speakers, they were a huge help to the virtual meeting. I am
thankful to acknowledge Dr. Periyathambi Dhaiveegan, Taiwan; Dr. Rajesh Kumar,
Chennai; Dr. Pradeep Kumar Panda, Taiwan; and Dr. Yusuf Sarkingobir, Nigeria for their
fruitful participation
cipation and presentation in the Nano
Nano-world
world science programme.
With Regards
Mrs. JANAKI S,
Assistant Professor and Head
Department of Microbiology
Vivekanandha Arts and Science College for Women, Sankari, Salem.
E.mail: janooravi@gmail.com
MESSAGE FROM EDITOR-IV
It’s my great pleasure and privilege to present the proceedings of the Virtual International
Conference on “ Advancements in Nanotechnology (VICAN – 2020), organized by Association of
Global Academicians and Research (AGAR) and Association of Indian Biologists (AIB) on
01.11.2020, with exciting and inspiring flow of in formations. The purpose of the conference is to
provide a forum for discussion on the applications of Nanotechnology towards the global needs,
and to analyze the practical challenges
allenges encountered and the solutions to be adopted.
The conference serve as an ideal platform for academicians, young researchers and students
from all over the country to share and update their innovative ideas and research experiences in all
diversifiedd fields of Nanoscience. I am assuring that this conference paves strategically pathway for
multidisciplinary research with high level of satisfaction and aspiration. This event includes paper
and poster presentations with awards for best presentation. Eac Eachh contributed paper was referred
before being accepted for publication in this proceeding. The papers were accepted for publication
based on their interest, relevance, innovation and application.
Really, I appreciate the authors for their contribution and the reviewers for their continuous
immense effort. Sincere thanks to the Keynote speakers, as they were a great asset to the virtual
conference. I am grateful to acknowledge Dr.Periyathambi Dhaiveegan, Taiwan; Dr. Rajesh
Kumar, Chennai; Dr. Pradeep Kumar P Panda,
anda, Taiwan and Dr.Yusuf Sarkingobir, Nigeria for their
effective involvement and presentation about the recent realms in the scientific program of Nano-
Nano
world. Great applause to the organizing committee members, press members and everyone who
contribute for
or the success of this conference with proceedings.
I hope that everyone had rewarding experiences to focus research in multi areas of Nanoscience. I
wish all the attendees of VICAN
VICAN-2020,
2020, an enjoyable, scientific virtual gathering with potential
resources.
Nano, being small…………
Enlighten with big…..miracles……
……….A Key To Future………………. With Regards
Mrs. ALBINO WINS J, J
Assistant Professor, Department of Botany
Holy Cross College (Autonomous),
Nagercoil - 4.
Kanyakumari District.
Mail ID: winsbt@gmail.com
MESSAGE FROM EDITOR
EDITOR-V
The Virtual International Conference on Advancements in Nanotechnology

organised by Association of Indian Biologists (AIB), Tamil Nadu. I am glad that all participants
took the opportunity to exchange their knowledge, experiences and ideas and also made contacts
and established further collaboration. This educational program, rich in events, provided more
relaxing atmosphere during the meetings among colleagues in this pandemic situation.
Indeed it is a privilege
ilege and honour to be part of the jointly arranged Virtual International
Conference on Nanotechnology Advances by the Association of Indian Biologists (AIB), Tamil
Nadu, organised with great magnitude in the COVID
COVID-19
19 era. Because of its enormous global
response,
sponse, this remains a reality. Indeed I am grateful to the members of the association for
providing me with an opportunity and for rebuilding faith in me with their assistance. My thanks to
the faculty members and the interested students. I'm so grateful to my associates for driving me in
need. Blessed and fortunate to be part of this prestigious gathering. I hope that we will meet at the
next conference VICAN with dynamic topics and more eminent expertise.
With Regards,
Ms. MUNEESWARI K,
Assistant Professor
Marudhar Kesari Jain College for Women
Vaniyambadi-635
Vaniyambadi 751
Tirupattur District, Tamilnadu.
E.Mail ID :recha27@gmail.com
MESSAGE FROM EDITOR
EDITOR-VI
I am delighted to know that the Association of Indian Biologists (AIB), Tamil Nadu
organized Virtual International Conference on Advancements in Nanotechnology (VICAN). My
hearty congratulations to the organizers, committee members and all the participants for their
untiring zeal and enthusiasm in imparting information to the students’ community.
A good education broadens our horizons and gives us good chances in life. Knowledge
enables people to grow and influence their situation. ““Education is the passport to the future, for
tomorrow belongs to those who prepare for it today.” “Your attitude, not your aptitude, will
determine your altitude.” “If you think education is expensive, try ignorance.” “The only person
who is educated is the one who has lea
learned how to learn and change.”
My sincere appreciation to the committee members for organizing this Virtual
International Conference on Advancements in Nanotechnology
Nanotechnology.. I am happy to wish the organizers
great success for the seminar.
With Regards,
Ms. P. AMUDHANILA,
AMUDHANILA
Assistant Professor,
Department of Biotechnology,
Marudhar Kesari Jain College for Women,
Vaniyambadi-635
Vaniyambadi 751
Tirupattur District, Tamilnadu.
E.mail: amudhanila.biotech@gmail.com
MESSAGE FROM ASSOCIATE EDITOR – I
Hello and welcome to Virtual International Conference on Advancements in

Nanotechnology (VICAN). It is my great pleasure to serve as conference chair for the first Virtual
International Conference on Advancements in Nanotechnology organized by Association of
Indian Biologists (AIB), Tamil Nadu. This conferences were lively events where education
researchers and practitioners from around the world came together to discuss a wide array of
important issues in education and research.
I am delighted that all participants took the opportunity to share their expertise,
perspectives and ideas, making connections and creating further cooperation. This event-rich
event
instructional curriculum provided more relaxing atmosphere during colleagues' meetings
m in this
pandemic situation. I hope during your time at the conference that you take the opportunity to
engage with your peers to discuss your ideas for research and practice and that you ask questions
of the presenters. There will be plenty of oppo
opportunities
rtunities for collaboration. We will all benefit from
our Virtual International Conference on Advancements in Research and Education.
With Regards,
Dr. KAMAL GULATI

Associate Professor,
Amity University, Noida, Main Campus,
Gautam Budh Nagar, Uttar Pradesh - 201 303
Email: drkamalgulati@gmail.com
MESSAGE FROM ASSOCIATE EDITOR - II
The conference serves as an outstanding venue for academics, aspiring experts, and students
around the country to explore and update their creative concepts and academic experiences across
diverse fields of nanoscience. This conference paves a theoretical path for multidisciplinary
research with strong contentment and aspiration. This event includes paper and poster presentations
and best-presentation
presentation honours. Eac
Eachh contributed paper was referred before accepting publication of
this proceeding. Publications were accepted based on interest, relevance, innovation and execution.
I am glad that all participants took the opportunity to exchange their knowledge,
experiences
es and ideas and also made contacts and established further collaboration. This
educational program, rich in events, provided more relaxing atmosphere during the meetings among
colleagues in this pandemic situation.
I hope that we will meet at the next conference VICAN with dynamic topics and more
eminent expertise.
With Regards,
Mr. MOHIT TIWARI

Department of Computer Science and Engineering,
Bharati Vidyapeeth’s College of Engineering,
New Delhi.
E-mail ID: mohit.tiwari@bharatividyapeeth.edu
MESSAGE FROM ASSOCIATE EDITOR – IIII
I am delighted to know that the Association of Global Academicians and Researchers

Rese
(AGAR), Andhra Pradesh and Association of Indian Biologists (AIB), Tamil Nadu organized
Virtual Internationall Conference on Advancements in Nanotechnology (VICAN).
(VICAN My hearty
congratulations to the organizers, committee members and all the participants for their untiring
zeal and enthusiasm in imparting information to the students’ community.
A good education broadens our horizons and gives us good chances in life. Knowledge
enables people to grow and influence their situation. ““Education is the passport to the future, for
tomorrow belongs to those who prepare for it today.” “Your attitude, not your aptitude, will
determine your altitude.” “If you think education is expensive, try ignorance.” “The only person
who is educated is the one who has learned how to learn and change.”
My sincere appreciation to the committee members ffor
or organizing this Virtual
International Conference on Advancements in Nanotechnology (VICAN).. I am happy to wish the
organizers great success for the seminar.
With Regards,
Ms. TRIPTI TIWARI
Department of Management Studies,
Bharati Vidyapeeth (Deemed to be University)
Institute of Management Research, New Delhi
E-mail ID: tripti.tiwari@bharatividyapeeth.edu
MESSAGE FROM ASSOCIATE EDITOR – IV
I am glad that all participants took the opportunity to exchange their information,
experiences and ideas and also made connections and formed further collaboration.
During the meetings between colleagues in this pandemic situation, this education
programme,
e, rich in activities, offered a more calming environment.
Good education spreads our horizons and gives us healthy life. Information
develops individuals and influences their condition. Training is the future visa, and
tomorrow belongs to us who are planning for it now. The only one educated has
h learnt to
learn and improve. I hope that we can meet with dynamic subjects and more eminent
knowledge at the next VICAN meeting.
With Regards,
Dr. T. ANANTH KUMAR

Assistant professor
Department of Computer Science and Engineering
IFET College of Engineering
Email ID: ananth.eec@gmail.com
MESSAGE FROM ASSOCIATE EDITOR – V
Science always strives to search, find, know, realize and prove. Technology tries to implement. Both
work for the benefits of not just the humanity but for the entire biological kingdom.
In the current scenario where COVID
COVID- 19 pandemic rules the destructive roost, biotechnology holds
the key in the resurrection of mankind. As we can say the 20th century was the era of electronics, similarly
si
21st century can be designated as the era of biotechnology. Biotechnology basically aims at improving the
quality of human life and at protecting him from dangerous diseases.
diseases.This
This book of Advanced Biological
Sciences (ABS) is likely to present the vvarious
arious facets in the form of research articles pertaining to Human
Health, Animal Health, Genetics, Nanobiotechnology, RDNA technology, Agriculture, Medicinal Plants,
Forestry Fisheries, Environment, Horticulture, Floriculture, Food processing, Renewable energy,
e
Aquaculture etc., The different articles that are to be presented in the book may pave way for further
advanced research, technological implementation and for industrial production if needed in a large scale.
This book will hence be of yeoman’s serv
service
ice for the entire spectrum of student, faculty, research scholars
and scientists. Nanobiotechnology, Herbal technology and RDNA technology are the three important areas
focussed and dealt in detail in this book.
My humble thanks to the peers in the Assoc
Association
iation of Indian Biologists (AIB), the authors, technical
crew, review committee and all who played a vital role in completion of this book with ISBN. Looking
forward for more such kind of insightful works from AIB.
With Regards,
Dr. A. SIVARANJINI,
Assistant Professor,
Department of Biotechnology,
D. G. Vaishnav College,
Chennai-
Chennai Tamilnadu.
Email ID – sivaranjinia@dgvaishnavcollege.edu.in
MESSAGE FROM ASSOCIATE EDITOR – VI
I am pleased to know that the Virtual International Conference on Advances

in Nanotechnology was organised by the Association of Indian Biologists (AIB), Tamil
Nadu (VICAN). My heartfelt congratulations to the organisers, members of the committee
and all the participants for their untiring energy and enthusiasm for sharing knowledge
with the student community.
Awareness helps individuals to develop and affect their condition. "Your attitude,
not your ability, will determine your altitude." "If you think education is costly,
costl try
ignorance." "The only person who is educated is the one who has learned how to learn and
change." "Education is the passport to the future, for tomorrow belongs to those who
prepare for it today."
My sincere gratitude to the members of the committe

committeee for organising this Virtual
International Conference on Nanotechnology Development. I am pleased to wish the
organisers of the seminar great success.
With Regards
Mr. A.UBAITHULLA
UBAITHULLA BAIG,BAIG
Assistant Professor in Physics
Department of Science & Humanities
C.Abdul Hakeem College of Engineering & Technology
Melvisharam
E.mail: baig.ubaidulla@gmail.com
CONTENTS
S.NO TITTLE PAGE NO.
PLANT-BASED GREEN APPROACHES OF IRON
NANOPARTICLES SYNTHESIS AND ITS BIOREMEDIATION
1
1 OF WASTEWATER
R. Siva Dharshini and M. Poonkothai
AN ASSESSMENT OF BACTERICIDAL ACTIVITY, CYTOTOXIC
EFFECT AND GREEN SYNTHESIS OF SILVER NANO-
PARTICLES FROM FRUIT COAT OF ANNONA RETICULATA
2 LINN. 9
D. S. Kharate, T. B. Namekar, P. S. Kharate, R. A. Satpute,
N. B. Pandhure, V. R. Lakwal, S. M. Salunke,
V. D. Suryawanshi, A. D. Adsare, S.S. Gade and M. S. Kharate
STRUCTURAL CHARACTERIZATIONS OF
NANOSTRUCTURED ZINC OXIDE THIN FILM GROWN BY
3 20
SPRAY PYROLYSIS TECHNIQUES
Zayyanu Shehu, Shi’itu Abubakar, Yusuf Sarkingobir and Niyas
Ahmed. M.I
ALGINATE AS MEDICAMENT IN PHARMACEUTICS
40
4 Ashwini Ravi, Hemapriya J and Vijayanand S
HARMFUL BIOLOGICAL EFFECTS OF PLASTICS By
63
5 Yusuf Sarkingobir and UmmuTukur
SURVEY ON WETLAND PLANTS OF PERIYAKULAM POND
IN VALLIOOR, TIRUNELVELI DISTRICT, SOUTH INDIA 75
6
J. Albino Wins, Selvanandhini.T and N.Nishanthi
NANOPLASTICS: SMALL SCIENCE WITH BIGGER
CONSEQUENCE
82
7 Yusuf Sarkingobir, Aminu Umar Imam and Kasimu Abubakar
Shagari
CORONAVIRUS PANDEMIC AND RELIEF PROCEDURES:
SUGGESTIONS FOR MATERNAL AND KID WELLBEING
91
8 AND NOURISHMENT
Shubham
APPLICATIONS OF BIOTECHNOLOGY – A REVIEW
103
9 S.Rasulmeera and Dr. R. Kungumapriya
APPLICATION OF BIOINFORMATICS TO
112
10 NANOTECHNOLOGY
Dr.K.Shoba
IMPORTANCE OF MICROBES IN PHARMACEUTICALS
11 118
Dr.K.Shoba and J. Buenefa Shalom Trephosa
ACQUISTION OF ELECTROSPIROGRAM
12 125
Chetana Krishnan
SYNTHESIS OF SKIN PROTECTANT FABRIC BY COMPLEX
13 MIXTURE OF FISH COLLAGEN AND CELLULOSE NANO
135
CRYSTALS FROM CASSAVA EXTRACT
V. Kaviya, Dr.Sr.G.S.Mary fabiola and V.Ilakkiya
APPLICATION OF NANOTECHNOLOGY IN CANCER
BIOLOGY 147
14
Dr. K. Shoba and Ms. K. Sathvika
NANO DRUG DELIVERY SYSTEM FOR THE TREATMENT OF
CARDIOVASCULAR DISEASES 154
15
Aakancha Shaw
GENE ANNOTATION AND PHYLOGENETIC ANALYSIS OF
CARCININ PROTEIN FROM CARCINUS MAENAS 157
16
Dr. K. Shoba and J. Buenefa Shalom Trephosa
EVALUATION OF DRUG SUSCEPTIBILITY WITH ALAMAR
BLUE IN MYCOBACTERIUM TUBERCULOSIS 170
17
J. Albino Wins and M. Murugan
GREEN FABRICATION OF HIGH PERFORMANCE ZN-CO
FILM THROUGH SUPERCRITICAL CO2
ELECTRODEPOSITION PROCESS: ITS CORROSION
EVALUATION 175
18 Manickaraj Shobana Sebastin Mary, Pandiyarajan Sabarison,
Muthuraja Kalpana Devi, Sheng-Tung Huang and Ho-Chiao
Chuang
A STUDY ON THERAPEUTIC POTENTIALS AND
MOLECULAR DOCKING OF PLANT EXTRACTS AGAINST
19 ENVIRONMENTAL ISOLATES CAUSING URINARY TRACT 177
INFECTION
Venkat S, Jayasree A, Pavithra M, Shoba G and Sivaranjini A
PHYTOCHEMICAL AND ANTIOXIDANT ANALYSIS OF
MEDICINAL PLANTS AND ITS ANTIBACTERIAL EFFECT ON
178
BACTERIAL PATHOGENS
20
M.K. Arya, Sathish D, Jayasree A Sivaranjini A, Pavithra M
NANOBIOREMEDIATION
179
21 Abirami S
HIRSCHSPRUNG INFECTION, RELATED DISORDER, AND
22 HEREDITARY QUALITIES: A SURVEY 182
Shubham
DEVELOPMENTAL HEREDITARY QUALITIES OF PLANT
23 VARIATION 183
Shubham
A POPULACE HEREDITARY QUALITIES (GENES)
PERSPECTIVE ON CREATURE TAMING 184
24
Shubham
SNPS IN LEGAL HEREDITARY QUALITIES: A SURVEY ON
SNP COMPOSING STRATEGIES 185
25
Shubham
METASTASIS QUALITIES: A MOVEMENT PUZZLE
186
26 Shubham
FACILE SYNTHESIS OF FES2/AG2MOO4 NANO-
HETEROSTRUCTURE FOR PHOTOCATALYSIS AND
189
27 BACTERICIDAL APPLICATIONS.
Kokilavani and S. Sudheer Khan
NOVEL GRAPHENE QUANTUM DOTS: A REVIEW ON
SYNTHETIC TECHNIQUES AND APPLICATIONS 190
28
Sridhar Arelli, Akshay.S and Niti Chawla
ADVANCEMENT OF IN SILICO APPROACHES FOR PROTEIN-
PROTEIN INTERACTION 238
29
Sugumari Vallinayagam, Kavita Khatana
30 AN ECO FRIENDLY CONCEPT OF GREEN COMPUTING
Dr. Sriram E 302
MAGNETO-ELECTRONIC PROPERTIES OF GRAPHENE
NANOPARTICLES 308
31 Sridhar Arelli and Akshay.S
A DETAILED REVIEW ON WOUND DRESSING APPLICATION
OF CHITOSAN BASED BIOMATERIALS 322
32
Pradeep Kumar Panda, Pranjyan Dash and Zoyeb Mohamed Zia
A COMPREHENSIVE REVIEW ON NANOPHOTONICS AND
ARTIFICIAL INTELLIGENCE 376
33
Sridhar Arelli and Srivenkaiahppalaswamy B
EVALUATION OF CHITOSAN-BASED BIOMATERIALS AS
34 BIOMEDICAL APPLICATIONS 428
K.Anu and NitikaTomar
ANTIMICROBIAL ACTIVITY OF PLANT-MEDIATED
SYNTHESIS OF SILVER NANOPARTICLES 446
35 S.D.K.Shri Devi, Sudha and Monika
MACHINE LEARNING IN DRUG REPURPOSING
36 479
Kiruthika G and Dr. Vasna Joshua
WOMEN’S HEALTH
480
37 Salma Suqlain M
DNA FIX
482
38 Shubham
“A WOMAN’S HEALTH IS HER CAPITAL”

THE RELATIONSHIP BETWEEN SYMPTOM BURDEN AND
SYSTEMIC INFLAMMATION DIFFERS BETWEEN MALE AND 488
39
FEMALE ATHLETES FOLLOWING CONCUSSION.
Sowmiya. N,
A REVIEW ON THE APPLICATION OF MICROBES IN
FORENSICS 490
40
Haritha S
AN EMPIRICAL STUDY ON FAMILY PLANNING: THE
HOUSEHOLD CONSUMPTION AND SIZE 498
41
Dr.Pralay Ganguly
LAWS AND ACTS FOR ENVIRONMENTAL PROTECTION
506
42 S.Rasulmeera and Dr. R. Kungumapriya
A COMPREHENSIVE REVIEW ON THE NANO- CONFINED
AND ENGINEERED NANOWOOD MATERIALS FOR WATER
43 512
PURIFICATION PROCESS
V. Ragul and M.I. Niyas Ahamed
POLYMORPHISMS IN DNA REPAIR GENES IN CARCINOMA
44 PATIENTS LIVING IN A VERY COAL-MINING REGION 529
Shubham
MEDICINAL TREE - Couroupitaguianensis

45 530
Vino Udappusamy
ARTIFICIAL INTELLIGENCE IN AGRICULTURE: A MINI

46 REVIEW 531
Atishya Mahesh Jain and Murugan S
GREEN CLOUD COMPUTING AND HEALTH CARE

47 538
Sylvia Grace J
TRANSGENIC ANIMALS
48 539
Nabiha zainab
PRODUCTION OF HUMAN INSULIN FROM rDNA

49 TECHNOLOGY 540
Nabiha zainab
GENOME EDITING
50 542
Nabiha zainab
MELANIN PIGMENTATION IN PEOPLE: INTRODUCTION,
51 TYPES, HEREDITARILY JOB, PRODUCTION AND FUNCTION 544
Shubham
DNA REPAIR IN MAMMALIAN CELLS ON EARTH

52 546
Shubham
HEALTH BENEFITS OF EATING TRADITIONAL FOODS

53 548
S.Rasulmeera and Dr. R. Kungumapriya
MICE WITH DIABETES SUCCESSFULLY TREATED WITH

54 ELECTROMAGNETIC FIELDS 549
R.Jumail Ahamed
SYNTHESIS OF CERIUM OXIDE NANOPARTICLES
FROMDIFFERENT MEDICINAL PLANTS AND THEIR
55 551
ACTIVITIES AND ITS MEDICINAL APPLICATION -A REVIEW
K.Sheela
STUDY ON STRENGTH PROPERTIES OF CEMENT CONCRETE
56 PAVER BLOCKS MADE WITH SILICA FUME – A REVIEW 552
S.Gunasekar & Dr.N.Ramesh
A REVIEW ON THE BIOSYNTHESIS OF NANOPARTICLES BY
57 MARINE ORGANISMS 557
Abinaya D, Brita John C and Dhivya P
MOLECULAR DOCKING STUDIES OF BIOACTIVE
COMPOUNDS FROM CENTELLA ASIATICA WITH BETA -
58 LACTAMASE OF PSEUDOMONAS AERUGINOSA: A 579
COMPUTATIONAL APPROACH
K.Raviya Gani, Sivaranjini Annamalai and Shoba Gunasekaran
SYNTHETIC TECHNIQUES IN NANOBIOLOGY
59 Pranjyan Dash, Pradeep Kumar Panda and Sridhar Arelli 580
PHYTOCHEMICALS AND PHARMACOLOGICAL

60 APPLICATIONS OF PLANTS 630
Poongothai Annadurai
PROCEEDINGS OF VIRTUAL INTERNATIONAL CONFERENCE ON ADVANCEMENTS IN NANOTECHNOLOGY [VICAN]
PLANT-BASED GREEN APPROACHES OF IRON NANOPARTICLES

SYNTHESIS AND ITS BIOREMEDIATION OF WASTEWATER
R. Siva Dharshini, M. Poonkothai
Molecular Genetics Laboratory, Department of Genetic Engineering, SRM Institute of Science
and Technology, Kattankulathur - 603203, Tamil Nadu, India
Department of Zoology, Avinashilingam institute for Home Science and Higher Education for
Women, Coimbatore- 641043, Tamil Nadu, India
Email ID - poonkothaiadu@gmail.com
ABSTRACT
Nanotechnology was a growing technology with versatile applications. Apart from
nanotechnology's physical and chemical approaches, biological approaches are highly studied
and preferred by researchers and industries. Plant-based metal nanoparticle synthesis was cost-
effective and highly flexible for many applications. Among metal nanoparticles, iron was
favorable for its unique sustainability and other features. Here, chemical and organic solvents for
the production of iron nanoparticles are replaced by the phytochemicals and aqueous matrixes.
Plant mediated synthesis is a convenient and straightforward process as same as any other
chemical or biochemical reactions. The unique feature of iron nanoparticles, such as adsorption
and compatible nature favors the bioremediation of wastewater. This review illustrates the plant-
based biosynthesis of iron nanoparticles and potential applications towards bioremediation of
wastewater.
KEYWORDS: Iron nanoparticles; plant; bioremediation; nanotechnology
INTRODUCTION
Nanotechnology is trending and advantageous in manufacturing and manipulating things on
the atomic scale (between 1 to 100 nm). These nano-scale ranged products are widely used in
mechanics, optics, electronics, biotechnology, microbiology, environmental remediation,
medicine, numerous engineering fields, and material science (Christian et al. 2008). Majorly,
nanoparticles are synthesized by two different approaches, such as the top-down approach and
the bottom-up approach. Both of these approaches are used to employ physical and chemical
methods. These production methods involve the active participation of toxic compounds such as
ISBN: 978-81-948129-1-3 Page 1

sodium borohydride and hydrazine hydrate. These toxic substances are dangerous to the
environment and public health (Virkutyte and Varma 2013). Several studies and trials are going
on to tackle those disadvantages and replace them with eco--friendly approaches. At the same
time, we have to be concerned about the reliability, sustainability and adaptability of
nanoparticles and the wide availability of sources. These technologies should balance economic
affordability and environmental sustainability (Jebali et al. 2011). Green methods of nanoparticle
synthesis include plants, microorganisms, and biological wastes-based nanoparticle production.
In green synthesis, active natural components such as enzymes work as reducing and as capping
agents for nanoparticle production on a large scale. The extracts from biological sources are used
to reduce the metal ions. Plant extracts are highly preferable because of their reducing and
stabilizing agents (Fahmy and Mamdouh 2018).
Iron was the most ubiquitous of the transition metals and the fourth most dominant element in
the Earth's crust. It was also the backbone of our infrastructure. Fe nanoparticles possess high
magnetic nature, surface area, electrical and thermal conductivity. Iron nanoparticles have many
advantages and applications in various fields of spintronics, biology, and biomedical science.
Ferro or ferromagnetic materials below the size of 10 to 20 nm exhibit superparamagnetism. It
was a unique feature present only in iron-based nanoparticles (Pankhurst et al. 2009). Among
the green approach to iron nanoparticle synthesis, plant-based biosynthesis is manageable,
adaptable, and decreased time-consuming. It was a cost-effective and straightforward process
(Dhillon et al. 2012). Many studies illustrated that plants are representatives for producing metal
nanoparticles using different parts of the plant: leaf, stem, seed, and root (Iravani 2011). The
presence of antioxidants and phytochemicals in the plants quickly adapt to the synthesis and
architecture of nanoparticles. These green approaches are pollution-free, sustainable, and clean
technology, among other green technologies (Katti et al. 2009).
Bioremediation of wastewater is an essential step to initiate to control the increasing water
pollution. The contaminants, such as phosphorous and nitrogen in water bodies, affect both
environment and human health (Malison et al. 2014). Many processes are already in practice to
eliminate water pollution. Those present processes have both advantages and disadvantages in all
aspects. Still, researchers are seeking a practical water treatment approach to improve the
environment. A symbiosis of bioremediation of water and nanotechnology was the potential and
eco–friendly technology (Stefaniuk et al. 2016). The properties of adsorption, chemical inertness,
ISBN: 978-81-948129-1-3 Page 2
biocompatibility, and superparamagnetism allow the iron nanoparticles to apply bioremediation.

Fe nanoparticles are nano absorbents for removing dyes and environmental contaminants (Liu et
al., 2009). This review discussed the combination of plant-based green synthesis of iron
nanoparticles and its potential bioremediation of water.
PLANT-BASED BIOSYNTHESIS OF IRON NANOPARTICLES

In this production technique, the plant extract and one of the iron salts (FeCl 3·6H2O,
FeCl2·4H2O, FeSO4, or Fe (NO3)3) are used as the iron precursor. The phytochemicals present in
the plant extract facilitates the conversion of iron ions to iron nanoparticles. Iron nanoparticles
include zero-valent (ZVI), ferrous ferric oxide, ferric oxide, iron oxide hydroxide, and iron
mineral complex nanoparticles.
Zero- valent nanoparticles are highly reactive by the presence of incompletely filled d- orbitals.
ZVI nanoparticles have high remediation properties in various environmental and industrial
processes (Ebrahiminezhad et al. 2017). Many different plant extracts are used to produce ZVI
nanoparticles and are widely used for wastewater bioremediation. Hoag et al. (2009)prepared
ZVI nanoparticles using tea leaf extract and applied them for the degradation and removal of
organic contaminants such as bromothymol blue. Azadirachta indica leaf extracts are used to
produce ZVI nanoparticles (Pattanayak and Nayak 2013). Machado et al. (2013)experimented
with the leaf extracts of different trees such as apple, apricot, avocado, cherry, eucalyptus, kiwi,
lemon, mandarin, medlar, mulberry, oak, olive, orange, passion fruit, peach, pear, pine,
pomegranate, plum, quince, raspberry, strawberry, black tea, green tea, vine, and walnut. The
ZVI nanoparticles were prepared using S. jambos, and Alston extract was reported to remove
cadmium (Xiao et al. 2016).
Magnetic iron nanoparticles have unique magnetic properties and ideal for biotechnological
and environmental remediation (Reguyal et al., 2017). Plant extract of Hordeum vulgare, Rumex
acetosa (Makarov et al. 2014), eucalyptus (Zhuang et al. 2015), green tea, and black tea (Kuang
et al. 2013) are reported for the production of magnetic iron nanoparticles. Iron oxide
nanoparticles are crystal in structure with different structural and magnetic properties (Machala
et al. 2011). The chestnut tree, eucalyptus, Gorse, pine, and black tea extracts are used to produce
iron oxide nanoparticles (Huang et al. 2014). As per application and plant extracts, the synthesis
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environment, such as pH, temperature, reducing agent, etc., differs for the appropriate size and
activity.
BIOREMEDIATION OF WASTEWATER USING IRON NANOPARTICLES

Photodegradation by nanoparticles is also common, and many nanomaterials are utilized for
this purpose (Rogozea et al., 2016). The removal of metal ions such as mercury, lead, thallium,
cadmium, and arsenic from natural water has attracted considerable attention because of their
adverse effects on environmental and human health. Iron oxide nanoparticles are useful sorbent
material for this toxic soft material. So, no measurements of engineered nanoparticles in the
environment have been available due to the absence of analytical methods to quantify trace
concentrations of nanoparticles (Mueller and Nowack, 2008). Apart from all iron nanoparticles,
zero-valent iron nanoparticles are advisable to remediate water due to their catalytic properties
(Machado et al. 2015). The ZVI NPs are adequate for the water treatment for the removal of
contaminants such as azo dyes (Poursaberi et al., 2012), brominated organic compounds, anti-
biotic, pesticides, nitrate, alkaline-earth metals, malachite green, monochlorobenzene, and
transition metals such as copper, chromium, and cobalt (Ebrahiminezhad et al., 2017).
The ZVI NP produced from plant extracts enhance the functionality of the nanoparticles
(Herlekar et al., 2014, Markarov et al., 2014). ZVI NPs from Shirazi thyme leaf extract and
Pistachio green hulls were applied to remove phosphorous (Soliemanzadeh et al., 2016). Zero-
valent iron oxide core-shell nanoparticles from green tea extract and eucalyptus exhibited nitrate
removal in swine wastewater (Wang et al., 2014). The iron oxide nanoparticles synthesized using
Amaranthus spinosus showed the degradation of methylene blue and methylene orange
(Muthukumar and Matheswaran, 2015). The degradation of organic dyes such as congo red,
methylene blue, and methyl orange are displayed by magnetite nanoparticles using Andean
blackberry leaf extract (Kumar et al., 2016). Hoag et al. (2009) reported ZVI nanoparticles using
tea leaf extract and applied for the degradation and removal of organic contaminants such as
bromothymol blue. The ZVI nanoparticles synthesized by using S. jambos and Alston extract
reported for the removal of cadmium (Xiao et al., 2016).
As mentioned above, many studies are focused on applying plant-based iron nanoparticles in
the bioremediation of wastewater. Even though the phytochemicals in plant extract improve
nanoparticles' stability, protein and antioxidant content parameters need to be measured. Several
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intensive studies are recommended for the remediation of wastewater by different iron
nanoparticles.
CONCLUSION
In this review, we focused on plant-based iron nanoparticles' ability to apply water treatment
and the biodegradation of contaminants. It displayed the merits of plant-based nanoparticle
synthesis over the physical and chemical methods. Even plant-mediated nanoparticles also have
limitations and demerits, like the flexibility of shape and monodispersity. These are
recommendations for the prospects of plant-mediated nanoparticles for practical applications and
increased bioremediation potential.
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AN ASSESSMENT OF BACTERICIDAL ACTIVITY, CYTOTOXIC EFFECT AND

GREEN SYNTHESIS OF SILVER NANO-PARTICLES FROM FRUIT
COAT OF ANNONA RETICULATA LINN.
D. S. Kharate, T. B. Namekar, P. S. Kharate, R. A. Satpute, N. B. Pandhure, V. R. Lakwal,
S. M. Salunke, V. D. Suryawanshi, A. D. Adsare, S.S. Gade and M. S. Kharate
Department of Zoology, Sant Ramdas Art’s, Commerce and Science College, Ghansawangi,
Jalna-431209,
Animal Physiology Laboratory, Department of Zoology, Dr. Babasaheb Ambedkar Marathwada
University, Aurangabad-431004
Department of Plant Molecular Biology and Biotechnology, Indira Gandhi Krishi
Vishwavidyalaya, Raipur -492 012
Department of Botany, Government Institute of Science, Aurangabad-431004
Plant Tissue Culture Laboratory, Department of Botany, Dr. Babasaheb Ambedkar Marathwada
University, Aurangabad-431004
Department of Biotechnology, Vinayakrao Patil College, Vaijapur, Aurangabad-423 701
P.G. Department of Zoology, Nanasaheb Yashwantrao Narayanrao Chavan Art’s, Science and
Commerce College, Chalisgaon, Jalgaon-424101
Department of Botany, Art’s, Commerce and Science College, Maregaon, Yavatmal-445303
Department of Environmental Science, Dr. Babasaheb Ambedkar Marathwada University,
Aurangabad-431004
Department of Botany, Vinayakrao Patil College, Vaijapur, Aurangabad-423 701
*Corresponding Email: mangesh.kharate121@gmail.com
ABSTRACT
Nanomaterial obtained by green synthesis technologies have been widely studied in recent
years owing to constitute cost-effective and environmental-friendly methods. In addition, there
are several works that report the simultaneous performance of the reducer agent as a
functionalizing agent, modifying the properties of the nanomaterial as a simple and economical
synthesis methodology. In present investigation, the synthesis of silver nanoparticles from of
Annona reticulata, were studied for bactericidal activity against pathogens by agar cup well plate
method using different dilutions such as 15 mg/ml, 20 mg/ml, 25 mg/ml, 30 mg/ml, 35 mg/ml
ISBN: 978-81-948129-1-3 Page 9

and 40 mg/ml. The fraction of extracts showed maximum activity at 25 mg/ml. Among the
pathogens, the maximum zone of inhibition was noted against Escherichia coli (MTCC 7040)
[36.2mm]; Salmonella typhi (NCTC 8394) [24.1mm]; Bacillus subtilis (ATCC 6051) [33.9] and
Staphylococcus aureus, (Isolated) (Wound infection, Pneumonia) [19.7mm]. These results
showed that the silver nanoparticle from Annona reticulata is an effective bactericidal agent. The
cytoxicity was also done by hemolytic assay.
KEYWORDS: Bactericidal Activity, Cytotoxic Effect, Silver Nano-particles, Annona

reticulata.
INTRODUCTION
Green synthesis of silver nano-particles are an ecofriendly and biocompatible process
[1], generally accomplished by using a capping agent/stabilizer (to control size and prevent
agglomeration), plant extracts, yeast, or bacteria [2]. In contrast to microorganisms, plants have
been exhaustively used, as apparent from Table- 1 [3]. This is because plant phytochemicals
show greater reduction and stabilization [4]. Eugenia jambolana leaf extract was used to
synthesize AgNPs that indicated the presence of alkaloids, flavonoids, saponins, and sugar
compounds [5]. Bark extract of Saraca asoca indicated the presence of hydroxylamine and
carboxyl groups [6]. AgNPs using leaves of Rhynchotechum ellipticum were synthesized, and the
results indicated the presence of polyphenols, flavonoids, alkaloids, terpenoids, carbohydrates,
and steroids [7].
Hesperidinwas used to form AgNPs of 20–40 nm [8]. Phenolic compounds of pyrogallol and
oleic acid were reported to be essential for the reduction of silver salt to form NPs [9]. Pepper-
leaf extract acts as a reducing and capping agent in the formation of AgNPs of 5–60 nm. It is
important to note that the effectiveness of many species of medicinal plants depends on the
supply of active compounds. It has been reported that most of the biologically active constituents
of extracts, such as flavonoids, tannins, and terpenoids, are highly soluble in water, but have low
absorption, because they are unable to cross the lipid membranes of the cells, have excessively
high molecular size, or are poorly absorbed, resulting in loss of bioavailability and efficacy.
Therefore some extracts despite having very good activity in vitro which are not reproducible in
vivo, are not used clinically because of these obstacles [10]. It has therefore been widely
proposed to combine herbal medicine with nanotechnology, because nanosystems can deliver the
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bioactive components at a sufficient concentration during the entire treatment period, directing
them to the desired sites of action, and hence potentiating the action of the compounds, an aspect
that conventional herbal treatments do not meet [10, 11]. Nanoparticles are materials that are
small enough to fall within the nanometric range, with at least one of their dimensions being less
than a few hundred nanometers. This reduction in size brings about significant changes in their
physical properties with respect to those observed in bulk materials. A very interesting
application of nanoparticles in the scope of life sciences is their use as ‘smart’ delivery systems
where they are usually loaded with a drug or therapeutic agent [12].
The various developed chemical and mechanical methods of producing nanoparticles include
ball milling, thermal quenching, precipitation techniques, vapor deposition. However, these
methods are often costly, and may result in toxic byproducts [13, 14, 15, 16 and 17]. The
commercial significance of nanoparticles therefore remains limited by the nanoparticle synthesis
process, which is generally energy intensive or requires toxic chemical solvents and is costly.
Biological approaches, including use of microorganisms or plant extracts to synthesize metal
nanoparticles, have been suggested. An emerging field in nanotechnology is the synthesis of
metal nanoparticles using herbal plants. Metal nanoparticles display improved and / or novel
properties compared to their source materials. These properties may be derived from their size,
morphology, or distribution. This method is referred to as the green approach and is
environmentally friendly.
Thus, the advancement of green syntheses of nanoparticles is progressing as a key branch of
nanotechnology; where the use of biological entities like microorganisms, plant extract or plant
biomass for the production of nanoparticles could be an alternative to chemical and physical
methods in an ecofriendly manner[18]. Among several noble metal nanoparticles, silver
nanoparticles have attained a special focus [18 46]. Silver nanoparticles (AgNPs) are of
particular interest because of their antimicrobial, anticancer and cytotoxic activities. We
previously reported about the successful synthesis of silver nanoparticles from ethanolic extracts
of fruits of Annona muricata [19], nevertheless, there had been no reported method or
publication on the use of ethanolic extracts of Annona muricata to prepare nanoparticles from
leaves of this plant, despite their known therapeutic potential. The aim of this study was
therefore to optimize a method for the synthesis of AgNPs from ethanolic extracts of leaves of
Annona muricata as well as to characterise the green synthesized AgNPs. Nanotechnology
ISBN: 978-81-948129-1-3 Page 11
explores a variety of promising approaches in the area of material sciences on a molecular level,
and silver nanoparticles (AgNPs) are of leading interest in the present scenario. This review is a
comprehensive contribution in the field of green synthesis, characterization, and biological
activities of AgNPs using different biological sources.
Biosynthesis of AgNPs can be accomplished by physical, chemical, and green synthesis;
however, synthesis via biological precursors has shown remarkable outcomes. In available
reported data, these entities are used as reducing agents where the synthesized NPs are
characterized by ultraviolet-visible and Fourier-transform infrared spectra and X-ray diffraction,
scanning electron microscopy, and transmission electron microscopy. In the present work, the
synthesis of silver nanoparticles from of Annona reticulata, were studied for bactericidal activity
against pathogens by agar cup well plate method. The maximum zone of inhibition was noted
against Escherichia coli; Salmonella typhi; Bacillus subtilis and Staphylococcus aureus. These
results showed that the silver nanoparticle from Annona reticulata is an effective bactericidal
agent. The cytoxicity was also done by hemolytic assay.
MATERIALS AND METHODS
PLANT MATERIALS:
The Fruit coat of Annona reticulata was collected from Aurangabad district, Maharashtra
India, where it is cultivated it as a medicinal plant. The plants were collected during the August-
2019 to December 2019 while flowering and fruiting. The materials were dried under shade and
after optimum drying, coarsely powdered and stored in well closed container till further use [20,
21 and 22].
TEST PATHOGENS
The Test pathogens, Escherichia coli; Salmonella typhi; Bacillus subtilis and Staphylococcus
aureus were obtained from Microbial Type Culture Collection and Gene Bank, Institute of
Microbial Technology, Chandigarh (PB) India and were sub cultured and maintained in Animal
Physiology Laboratory, Department of Zoology, Dr. Babasaheb Ambedkar Marathwada
University, Aurangabad-431004 [23, 24, 25 and 26].
ISBN: 978-81-948129-1-3 Page 12

PREPARATION OF MICRO ORGANISM:

A loop full culture of pure strains of different pathogens were inoculated into 100 ml of
sterile Lowenstein Jensen media and incubated for four days at 37º C for bacterial culture. After
four days of incubation, 0.5 ml of broth containing the microorganisms were added into 9 ml of
Lowenstein Jensen media. The Ten fold serial dilutions were made in the range of 10 –1 to 10–9.
100 μl of the dilutions ranging from 10–5 to 10–8 were spread on the sterile Lowenstein Jensen
media plates and kept at 37º C for four days. The number of colonies forming units was counted
and numbers of microorganisms in each ml of stock culture were calculated [27].
PREPARATION OF PLANT EXTRACTS

The fruit coat of Annona reticulata, without any infection was collected and 50 g of the
leaves were weighed and washed with double-distilled water before use. The leaves were air-
dried for 10 days and were then kept in the hot air oven at 60ºC for 24–48 h. The leaves were
then cut into fine pieces and 100 ml of double-distilled water was added. The mixture was boiled
for 10 min before being decanted, and then was cooled and filtered through Whatman No. 1 filter
paper. The boiled extract was refrigerated and used for further experimental procedures [28].
GREEN SYNTHESIS OF SILVER NANOPARTICLES

Green synthesis of silver nanoparticles was done according to the method of Song and Kim
[29, 30]. The collected samples air dried for 10 days and were kept in the hot air oven at 60ºC for
20 minute. The leaves were ground to a fine powder. 1 mM silver nitrate solution was added to
plant extract to make up a final volume of 200 ml and centrifuged at 18,000 rpm for 25 min. The
collected pellets were stored at 4 C. The supernatant was heated at 50º–95º C. A change in the
color of solution was observed during the heating process [30].
UV-VISIBLE SPECTROSCOPY ANALYSIS

The color change in reaction mixture (metal ion solution + Annona reticulata extract) was
recorded through visual observation and Silver nanoparticles were characterized by UV–vis
schimadzu1600 spectrophotometer. The bioreduction was monitored and the absorption spectra
in 300–700 nm range. The bioreduction of silver ions in aqueous solution was monitored by
periodic sampling of aliquots (1 ml) and subsequently measuring UV-Vis spectra of the solution.
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BACTERICIDAL ACTIVITY
The bactericidal activity of silver nanoparticles from Annona reticulata was evaluated by cup
plate method using 15 mg/ml, 20 mg/ml, 25 mg/ml, 30 mg/ml, 35 mg/ml and 40 mg/ml dilutions.
Sterilized nutrient agar plates were prepared under aseptic conditions. Six mm diameter holes
were made in the agar plates using a sterile borer. 0.2 ml of the test organisms was spreaded on
Lowenstein Jensen media plates. Samples, standard drug penicillin were dissolved with DMSO
and the solvent control (DMSO) was added into each hole separately. The plates were
maintained at 4º C for 30min to allowed diffusion of solution into the agar medium. The plates
were incubated at 37ºC for one to four days for bacteria. The zone of inhibition was measured
using antibiotic zone reader [30].
HEMOLYTIC ACTIVITY
The hemolytic activities of the test compounds were determined using human red blood cells
[30]. Human erythrocytes from healthy persons were collected in tubes containing EDTA (1–2
mg/ml) as anti-coagulant. The erythrocytes were harvested by centrifugation (Heraeus Megafuge
40, Thermo Fisher Scientific Inc., MA) for 10 min at 634 × g at 20°C, and washed three times in
PBS. PBS was added to the pellet to yield a 10% (v/v) erythrocytes/PBS suspension. The 10%
suspension was diluted 1:10 in PBS.
From each suspension, 100 μL was added in triplicate to 100 μl of a different dilution series
of AgNPs in the same buffer in Eppendorf tubes. Total hemolysis was achieved using 1% Triton
X-100. The tubes were incubated for 1 h at 37°C and centrifuged for 10 min at 634 × g at 20°C.
From the supernatant fluid, 150 μl was transferred to a flat-bottomed micro titer plate (Himedia
Ltd., India), and the absorbance was Measured at 450 nm.
RESULTS AND DISCUSSION

The silver nanoparticles of Annona reticulata was found to be effective against various
bactericidal pathogens as indicated by the zone of inhibition. The fraction of extracts showed
maximum activity at 25 mg/ml. Among the pathogens, the maximum zone of inhibition was
noted against Escherichia coli (MTCC 7040) [36.2mm]; Salmonella typhi (NCTC 8394)
[24.1mm]; Bacillus subtilis (ATCC 6051) [33.9] and Staphylococcus aureus, (Isolated) (Wound
ISBN: 978-81-948129-1-3 Page 14

infection, Pneumonia) [19.7mm]. These results showed that the silver nanoparticle from Annona
reticulata is an effective bactericidal agent. The cytoxicity was also done by hemolytic assay.
Table 1: Bacterial activity of the AgNPs of Annona reticulataby cup plate method
Zone of inhibition in mm
Sr.
Test pathogens 15 20 25 30 35 40
No.
mg/ml mg/ml mg/ml mg/ml mg/ml mg/ml
01 Escherichia coli (MTCC 7040) 30.1 35.9 36.2 33.4 15.5 19.7
02 Salmonella typhi (NCTC 8394) 19.7 22.6 24.1 23.9 21.4 26.7
03 Bacillus subtilis (ATCC 6051) 26.7 29.7 33.9 34.7 9.7 11.9
04 Staphylococcus aureus (Isolated) 11.9 13.4 19.7 14.2 17.6 12.5
Figure- 1: Bactericidal activity of the AgNPs of Annona reticulata
The in vitro hemolytic assay is a screening tool for developing in vivo toxicity towards host cells
[30].
ISBN: 978-81-948129-1-3 Page 15
The silver nano partials showed no significant toxicity to human erythrocytes at minimum
growth inhibitory concentrations (P= 0.0744 for AgNPs) only 0–14% hemolysis was observed at
the tested concentrations, while penicillin showed 100% hemolysis at 14 μg/mL. In future, there
has been a mounting interest in the discovery of new antimicrobial compounds due to a startling
increase in the rate of infections with antibiotic resistant microorganisms. There is a urgent need
to have green nano particles from plants those are non toxic, target oriented, cost effective, novel
agent. Bacillus subtilis is an infectious bacterial disease that can be transmitted from person to
person because of the droplets from the throat and lungs of people suffering from the active
respiratory disease. TB has remained a serious problem in developing countries, and the
multidrug resistant strains of Bacillus subtilis are imposing the restrictions on the usage of
current bactericidal drugs. Alarming data from the World Health Organization (WHO) revealed
that Bacillus subtilis has spread to every corner of the globe (WHO, 2005). Present investigation
shows that the green synthesis of silver nano particles from Annona reticulata are producing
novel drug against Bacillus subtilis those are responsible for inhibition of pathogens.
CONCLUSION
In the present study, silver nanoparticles from Annona reticulata plant showed significant
bactericidal activity against all test pathogens by agar cup plate method and this activity may be
attributed to the presence of novel safe, effective, less toxic and target oriented plant drug present
in the plant as antibacterial agent. The present work revel that potential application of silver
nanoparticles of Annona reticulata as an antibacterial agent.
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ISBN: 978-81-948129-1-3 Page 19
STRUCTURAL CHARACTERIZATIONS OF NANOSTRUCTURED ZINC OXIDE
THIN FILM GROWN BY SPRAY PYROLYSIS TECHNIQUES
Zayyanu Shehu , Shi’itu Abubakar , Yusuf Sarkingobir , Niyas Ahmed.M.I

Department of Physics, Shehu Shagari College of Education Sokoto, Sokoto state, Nigeria
Department of Biology, Shehu Shagari College of Education Sokoto, Sokoto state, Nigeria
PG and Research Department Biochemistry, Sacred Heart College (Autonomous), Tirupattur,
Tamil Nadu, India
Email: superoxidedismutase594@gmail.com
ABSTRACT
This paper explains how structured zinc oxide thin film was prepared by spray pyrolysis
technique and to analyze structural properties of the film. Nanostructured zinc oxide thin film
was deposited on a soda lime substrate by electric low cost spray pyrolysis machine equipped
with a deposition controller at a substrate temperature of 300K. The deposited films are annealed
at 400K. The X-ray diffraction machine shows that the films revealed polycrystalline and
confirmed the uniformity and well grown crystalline morphology of the ZnO films. The grain
size is 0.494nm, interplar spacing is 0.09nm and lattice parameter is 1.8 x 10 -4m. Although all
the deposited films possessed good characteristic for use in photovoltaic application such as
good crystallinity, larger grain size and an optimum dislocation density, film annealed under
nitrogen at 3000C was found to be the best and contains less impurity. There is possibility of
commercial production of the most suitable solar cells using zinc oxide as it possessed promising
structural properties and in addition , zinc oxide contains neither Tran metals nor touce material.
KEYWORDS: Nanostructured, Zinc Oxide, thin film, Spray pyrolysis, Techniques
1. INTRODUCTION
Atmospheric and environmental pollution as a result of extensive fossils fuel utilization in
almost all human activities has lead to some undesirable phenomena that have not been
experienced before in human history. These phenomena include global warming, greenhouse
effect, climate change, ozone layer depletion and acid rain (Sen, 2008). It has been understood
scientifically through different researches that these are closely related to fossil fuel used this is
because fossil fuel emit greenhouse gases such as carbon dioxide and methane which hinder long
ISBN: 978-81-948129-1-3 Page 20

wave terrestrial radiation from escaping into space and consequently the earth troposphere
becomes warmer (Ludwig, 2005; Sen, 2008). In order to avoid further impact of these
phenomena the two main alternative are either to improve the fossil fuel quality thus reducing
their harmful emission into the atmosphere or more significantly, to replace fossil fuel usage as
much as possible with environmental friendly, clean and renewable energy sources such as solar,
wind, geothermal, hydropower, biomes and tidal energy source. There has been an emergence of
interest in solar energy utilization principally due to the rising cost of energy from convectional
source such as fossil fuel(Chopra,1969; Hideaki, 2008; Mohan,2009).
Solar radiation by nuclear fusion reaction deep in the sun`s core. The sun provides all the
heat and light received by the earth. Solar energy is refers to as renewable energy because it will
be available as long as the sun continues to shine. Estimated for the life of the main stage of the
sun are another four to five billions years (2008). In four-fifth of the sun`s energy falls on oceans
and drives the water remaining one-fifth of the sun`s energy falls on land and is still about 2000
times greater than total present world energy demand (Adegoke et al., 2000). The two main
technologists that have been developed to capture this energy are: solar thermal which is the
conversion of solar radiation to heat energy and solar photovoltaic cell which convert solar
energy direct to electricity.
Silicon is the most commonly used material for the manufacturing of solar cells. Due to its
indirect band gap and low absorption coefficient, si-based cells are typically very thick leading to
significant increase in the module cost; therefore it is highly desirable to develop low-cost
photovoltaic materials with high absorption efficiency (2010).
Thin film solar cells offer the promise of both low cost and scalability features that are vital
for any approach towards providing large amount of carbon free power (Shannon et al.,
2009).The most successful thin film solar cells belong to the following materials family (i) CIGS
(copper indium gallium selenite) family, where the absorbing semiconductor consist of CuInS
e2, or related material or alloy, such as CuInS2, Cu (In, Ga) Se2 or even Cu (In, Ga) (Se, S) (II)
The CdTe family; and (iii) the silicon family, consisting of an amorphous or micro amorphous
silicon silicon-hydrogen alloy or related alloy (Barret, 1956, Ludwig, 2005).
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Chalcopyrite type semiconductor like Cu(InGa)S2 or Cu(In, GA)Se2 have very beneficial
properties for photovoltaic applications, which lead to strong scientific interest in thin film solar
cells based on these materials, however gallium and indium for the preparation of the active
layer are very rare and expensive elements. This could lead to a shortage in the supply of these
elements and would inhibit a cost-effective large-scale production (Achim, 2010). To overcome
these limitations, alternative materials are heavily researched in order to substitute the expensive
element indium and gallium.
Aiming to develop the solar cells free from environmental contaminants, promising solar
cells of thin film type could be produced by zinc oxide. This is obtained by growing zinc oxide
using spray pyrolysis. Zinc oxide thin films were deposited by spraying zinc acetate onto heated
glass substrate at 300 °c. Zinc oxide possessed promising optical properties, energy band gap and
large absorption coefficient in the order of 10*4cm*-1 which means that large possibility of
commercial production of the most suitable solar cells using ZnO film and in addition zinc oxide
contain neither rare metal nor toxic material (Chopra, 1956; Ajay, 2006; Patal et al, 2012). The
objective of this paper was to carry out structural characterizations of nanostructured zinc oxide
thin film grown by spray pyrolysis techniques.
2. MATERIALS AND METHODS
2.2.1 SELECTION OF SUBSTRATE

In this study soda lime glass of size and thickness were used as substrate material due to their
fulfillment of the most of the below requirements as good substrate (Smith., 2001). They are:
Flat and smooth surface, High mechanical strength to enable the substrate to withstand strain
during processing and monitoring, High resistivity, High thermal conductivity, Low cost, Zero
porosity to minimize out gassing and to ensure film uniformity.
2.2.2 CLEANING OF SUBSTRATE

The substrates were first washed with ordinary detergent solution and these were treated in a
mixture of nitric acid and ethanol. The substrates were then washed with freshly prepared
distilled water and rinsed with acetone. The substrates were put vertically in cleaned dishes so
that there is no water stick-mark on the substrates.
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2.2.3 PREPARING PRECURSOR SOLUTION

To obtain precursor solution for deposition 1.1g of zinc acetate were measured with
measuring cylinder and pour into a cleaned beaker. 15ml of distilled water were added into the
beaker.30ml of ethanol with 99.99% purity was also poured into solution. 5ml of acetate acid and
50ml acetone were accurately measured and poured into the solution. Allow the solution to cool
for about 10minutes. The solution obtained is called precursor solution and are ready to be used
for deposition (Ajay, 2006).
2.2.4 DEPOSITION OF PRECURSOR SOLUTION

Zinc oxide is deposited on glass substrates by electric low-cost spray pyrolysis machine
equipped with deposition controller. The following steps were taken to achieve the deposition of
zinc oxide;
i. 1.01ml of precursor solution was obtained by syringe chamber which will be tight in the
spraying chamber of machine.
ii. The spray chamber was then opened and 3 substrates were mounted vertically on the
substrate holder and additional substrate was mounted horizontally to support the
three and screwed the plunger at 11.0cm as the source to substrate distance.
iii. The chamber was then closed and heated.
iv. An automatic voltage of 17kva was set and kept constant throughout the experiment.
v. The electrostatic and heater were on until the temperature reached 300k.
vi. The precursor solution was sprayed for about 10 minutes when zinc oxide started
spraying and depositing on the substrate, flow rate was maintained at o.8ml/min.
vii. The electrostatic was off and the remaining solution was exhausted from the syringe, and
the make one complete run.
viii. The above procedure was repeated for about three times (Ajay, 2006; Mohan, 2009)
2.2.5 ANNEALING OF ZINC OXIDE LAYER

After spray pyrolysing, as-deposited ZnO thin films were removed from the spray
pyrolysing chamber and were immediately rapid thermal annealing under nitrogen at tube
ISBN: 978-81-948129-1-3 Page 23

furnace at various temperatures were increased from 3o0℃ to 400℃. The heating rate was 6℃/s
and the dwell time was 1 h. After annealing, the samples were taken out for characterization.
2.2.6 X-RAY DIFFRACTION STUDIES OF THE SAMPLES

X-ray diffract meter is a very useful analytical instrument for determination of whole
range of the detail information about crystal structure, orientation, crystal sizes, lattice constant,
defect, stress and strain in the sample. X-ray diffract meter was used to X-ray diffraction spectra
.X-ray diffraction analysis including the peak search was done by computer programming expert
software. XRD patterns of all the films as well as bulk samples were taken from 0° to 80°.
FIG. 3 Schematic representation of a Diffract meter
3.O RESULTS AND DISCUSSION
3.1.1 Deposition of precursor solution and annealing with Nitrogen
Table 4.1 prepared samples, deposition temperature and annealing temperature
S/N CODE SUBTRATE TEMPERATURE (°C) ANNEALING TEMPERATURE

1 ZnO-1 300 UNANNEALED
2 ZnO-2 300 300
3 ZnO-3 300 400
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FIG 3.1 X-RAY pattern of un-annealed ZnO
FIG 3.2 X-ray pattern of ZnO ANNEALED UNDER N2 AT 300°C
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FIG 3.3 ZnO ANNEALED UNDER NITROGEN AT 400°C
3.1.1 SAMPLE ONE (ZNO AS DEPOSITED)
Table 3.2 X-ray diffraction peak for Zn0-1 ITS corresponding ZnO powder diffraction file
ZnO-1 REFERENCE
2Ө Pos.[°2Th.] Height [cts] FWHM [°2Th.] D-spacing [Å] Rel.Int.[%]
31.7324 31.7464 414.70 0.3346 2.81869 100.00

56.5911 56.5903 108.20 0.5712 1.62506 26.o9
3.1.1.1. Interplaner spacing (d)
(i) 2d (1 0 0) sinӨ= ....................4.1
-10
𝜆=1.504Å=1.5 m
d (1 0 0) =𝜆/ (2sinӨ)
-10
d (1 0 0) = 1.5 / (2 sin 31.7464)
0.583n
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(ii) 2d (101) sinӨ=
-10
𝜆=1.504Å=1.5 m
d (101)=𝜆/(2sinӨ)
-10
d (101) = 1.5 / (2 sin56.5903)
0.2511nm
3.1.1.2 GRAIN SIZE (D)
Where βi=0.3346
Β ii=0.5712
……………….4.2
0.9
D
 cos 
-10
Di = (0.9 1.504 )/ (0.3346 cos 31.7464)
= 0.476nm
-10
Dii = (0.9 1.504 )/ (0.5712 cos 56.5903)
=0.4304nm
3.1.1.3 DISLOCATION DENSITY (Δ)
δ= ……………..4.3
Where n=1
δ i= 1/ (4.76 10-10)2
= 1018 line m-2
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δ ii= 1/ (4.304 10-10)2
= 1018 line m-2
3.1.1.4 Number of crystallites (N)
Thickness of the film=100nm
N D3/T ……………..4.4
Ni (0.476 10-9)3/100 10-9
0.108 10-22m2
Nii (0.4304 10-9)3/100 10-9
7.973 10-22m2
4.1.1.5 MICRO STRAIN (Ɛ)
…………..4.5
Ɛi = (0.3346 cos 31.7464)/4
= 71.13 10-3
Ɛii = (0.512 cos 56.5903)/4
=70.48 10-3
4.1.1.6 LATTICE PARAMETER
……………………4.6
The h k l values are: h2+ k2+ l2=3, where h=1,k=0,l=0,
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For 1 0 0
a2 = d2 (h2+ k2+ l2)
-9
= 28. )
a= =0.0000168
The h k l values are: h2+ k2+ l2=3, where h=1, k=0,l=1,

For 1 0 1
a2 = d2 (h2+ k2+ l2)
-9
= 16.2506 )
a= =0.000018
3.1.2 SAMPLE TWO (ZNO ANNEALED UNDER N2 AT 300°C)

Table 4.3 X-ray diffraction peak for Zn0-1 ITS corresponding ZnO-2 powder diffraction file
Zn0-2 REFERENCE
2Ө Pos. [°2Th.] Height [cts] FWHM [°2Th.] D-spacing [Å] Rel.Int. [%]
31.7172 31.7173 639.12 0.2342 2.82120 100.00
56.6247 56.6248 213.23 0.4896 1.62415 33.36
3.1.2.1 INTERPLANER SPACING (D)
(i) 2d (1 0 0) sinӨ= ………..4.6

-10
𝜆=1.504Å=1.5 m
d (100) =𝜆/ (2sinӨ)

-10
d (100) = 1.5 / (2 sin 31.7173)
1.58nm
(ii) 2d (101) sinӨ=
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-10
𝜆=0.1504Å=1.5 m
d (101) = 𝜆/ (2sinӨ)
-10
d (100) = 1.5 / (2 sin56.6248)
0.251nm
Where βi=0.2342
Β ii=0.4896
0.9
D
 cos 
-10
Di = (0.9 1.504 )/ (0.2342 cos 31.7173)
= 1.1nm
-10
Dii = (0.9 1.504 )/ (0.4896 cos 56.6248)
=0.502nm
n
δ=
D2
Where n=1
δ i=1 / (11 10-10)2
= 1018 line m-2
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δ ii= 1/ (5.20 10-10)2
= 1018 line m-2
3.1.2.4 NUMBER OF CRYSTALLITES (N)
N D3/T
Ni (1.1 10-9)3/100 10-9
133.1 10-22m2
Nii (0.502 10-9)3/100 10-9
12.65 10-22m2
3.1.2.5 Micro strain (Ɛ)

 cos

4
Ɛi = (0.2342 cos 31.7173)/ 4
= 49.8 10-3
Ɛii = (0.4896 cos 56.6248) /4
3
=67.33 10-
3.1.2.6 Lattice parameter
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For 1 0 0
a2 = d 2(h2+ k2+ l2)
-9
= 0.2821 )
a= =0.0000167
The h k l values are: h2+ k2+ l2=3, where h=1, k=0, l=1,
For 1 0 1
a2 = d2 (h2+ k2+ l2)
-9
= 0.162515 )
a= =0.0000127
3.1.3 SAMPLE THREE (ZNO ANNEALED UNDER N 2 AT 400°C)
Table 3.4 X-ray diffraction peak for Zn0-1 ITS corresponding ZnO-2 powder diffraction file
ZnO-3 REFERENCE
2Ө Pos. [°2Th.] Height [cts] FWHM [°2Th.] D-spacing [Å] Rel.Int. [%]
31.6334 31.6335 485.37 0.2342 2.82849 100.00
56.4745 56.4745 97.88 0.4896 1.62811 20.17
3.1.3.1 INTERPLANER SPACING (D)
(i) 2d (1 0 0) sinӨ=
-10
𝜆=1.504Å=1.5 m
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d (100) =𝜆/ (2sinӨ)
-10
d (100) = 1.5 / (2 sin 31.6333)
0.1433nm
(ii) 2d (101) sinӨ=
-10
𝜆=0.1504Å=1.5 m
d (101) =𝜆/ (2sinӨ)
-10
d (111) = 1.5 /(2 sin56.4745)
0.09nm
Where βi=0.2342
Β ii=0.4896
0.9
D
 cos 
-10
Di = (0.9 1.504 )/(0.2342 cos 31.6333)
= 0.685nm
-10
Dii = (0.9 1.504 )/(0.4896 cos 56.4745)
=0.494nm
n
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D2
δ=
Where n=1
δ i= 1/ (6.85 10-10)2
= 1018 line m-2
δii= 1/ (4.94 10-10)2
= 1018 line m-2
3.1.3.4 Number of crystallites (N)
N D3/T
Ni (0.685 10-9)3/100 10-9
32.14 10-22m2
Nii (0.494 10-9)3/100 10-9
12.06 10-22m2
4.1.3.5 MICRO STARIN (Ɛ)
 cos

4
Ɛi = (0.2342 cos 31.6333)/4
= 49.8 10-3
Ɛii = (0.4896 cos 56.6248)/4
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=49.85 10-3
3.1.3.6 LATTICE PARAMETER
The h k l values are: h2+ k2+ l2=3,where h=1,k=0,l=0,
For 1 0 0
a2 = d2 (h2+ k2+ l2)
= 0.8281 -9 )
a= =0.00002878
For 1 0
a2 = d2 (h2+ k2+ l2)
= 0.162811 -9 )
a= =0.000018
Table 3.6 summary for (1 0 0)
ISBN: 978-81-948129-1-3 Page 35

SAMPL GRAI INTERPLANA NO. OF MICR DISLOCATIO LATTICE

E N R SPACING CRYSTALLIT O N PARAMETE
NAME SIZE d (nm) ES m2 STRAI DENSITY(δ) Ra
D (nm) (N) 10-22 N Ɛ x1014lines/m2 ( m)
X10-3
Zn0 as 0.476 0.583 0.103 71.13 4140 1.68
deposite
d
Zn0 1.100 1.580 1333.1 49.80 8200 1.67

annealed
under N2
at 300°c
ZnO 0.655 0.1433 32.14 49.8 2130 1.8

annealed
under N2
at 400°c
Table 3.7 summary for (1 0 1)
SAMPL GRAI INTERPLANA NO. OF MICR DISLOCATIO LATTICE

E N R SPACING CRYSTALLIT O N PARAMETE
NAME SIZE d (nm) ES m2 STRAI DENSITY(δ) Ra
D (nm) (N) 10-22 N Ɛ x1014lines/m2 ( m)
X10-3
ZnO as 0.4504 0.2511 7.973 70.48 5598 1.80
deposite
d
ZnO 0.502 10.251 12.65 67.33 3698 1.27

annealed
under N2
at 300°c
ZnO 0.494 0.09 12.06 49.85 4098 1.8

annealed
under N2
at 400°c
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3.2 DISCUSSION
3.2.1 XRD PATTERN
The XRD patterns of the spray pyrolysis coated as- de-posited ZnO and annealed ZnO
films are shown in Figures above. The intensity data were collected over a range 2θ is 20 to 80º.
All the diffraction peaks were well indexed to the hexagonal zinc oxide wurtzite structure
.diffraction peak corresponding to impurity were not found in the XRD pattern confirming high
purity of the synthesized product The result indicates that the films have a polycrystalline
hexagonal wurtzite structure with (1 0 0) and (1 0 1) orientation. In particular (1 0 0) (annealed
under 300°c) oriental growth is the most prominent peak of zinc oxide structure that belong to
hexagonal wurtzite system. The XRD pattern of un-annealed ZnO thin film (Fig. (4.1)) shows
very weak (1 0 0) and (1 0 1) diffraction peaks. . Fig. 4.2 shows a strong X-ray peak from (1 0 0)
planes with a weak (1 0 1) peak at annealed 300℃. As the annealed temperature was increased
further to 400℃, the intensity of the (1 0 0) peak in pattern decreased in comparing with the
pattern Fig. 4.2 as shown by fig. 4.3 but (1 0 1) peak did not change. The (1 0 0) peak intensity is
increased and then decreased with increasing temperature over the selected annealing
temperature.
The average sizes of crystallites (D) were calculated from the full width at half maximum
(FWHM) of the diffraction peaks from the (1 0 0) and (1 0 1) planes of the films with the help of
Scherer formula.
The average crystallite sizes were calculated for the ZnO-1, ZnO-2 and ZnO-3 thin films,
respectively. Using XRD data, the lattice parameters (a and c) are be calculated from
1/d2=4/3[(h2+hk+k2)/a2] +l2/c2,
Dislocation density (δ) can be calculated from
δ = 1/D2
And the micro strain (ε) can be calculated from
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ε = (λ/ (Dsinθ)-(β/tanθ))
Which will elucidate the nature of the prepared ZnO films? The measured line-widths
and the other physical parameters are calculated and are given in Table 4.6 and 4.7 decreases in
dislocation density and strain is due to the manifestation of dislocation network on the film
surface. This formation of the films is high quality films. The average lattice parameters, micro
strain and inter planer spacing (d) are close and good agreement with the standard results.
4. CONCLUSION
Zinc oxide films have been successfully prepared on glass substrates using the spray
pyrolysis technique. The process parameters were optimized to have good quality crystalline
films. XRD characterization of both un-annealing and annealing films revealed poly-crystalline
and confirmed the uniformity and well grown crystalline morphology of the ZnO films. The
effect of annealing temperatures on structural properties of the deposited films were studied,
these revealed all the deposited films possessed good characteristics for use in photovoltaic
application such good crystallinity, larger grain sizes and an optimum dislocation density. Zinc
oxide thin film can be used as an absorber layer. Annealing temperature is an important
parameter that determines the quality of the deposited films. Film annealed under nitrogen at
300°c was found to be the best and contain less impurity.
5. RECOMMENDATION
Annealing temperature which is factor that influence the formation of impurity phased
should be within 300°c since the film annealed under nitrogen at 300°c showed most prominent
peak.
The film thickness determined from the value of deposition rate and deposition time which
are constant throughout the deposition of measuring it using profilometer which may have
instrumental error.
There is possibility of commercial production of the most suitable solar cells using zinc
oxide as it possessed promising structural properties and in addition zinc oxide contain neither
rare metals nor toxic materials.
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REFERENCES
1. Adegoke, C.W Oluwafisoye et al (2010) operational design parameters in a multi-lamp
module solar street light system solarsystem.Nigeria journal of solar energy. Vol.20.
pp1-8
2. Achim F. et al 2010) investigation of formation from metal salts and
thioacetamide. Chemistry of material article Vol.30. p2
3. Aicha A. R Elshabini, R. Fred D. (1998) thin film technology handbook, McGraw Hill,
New york. Pp121-123
4. Ajay K.S. (2006) an introduction to solid state electronic devices. Macmillan, India
Pp216-236
5. Barret C.S. (1956) structure of metals, crystallographic methods, principle and data. Mc.
GRAW Hill, New york. PP346-236
6. Chan, C.P., lam, H and Surya, C. preparation of films by electro deposition
using ionic liquids. Solar energy material and solar cells.
7. Chopra, K.L. (1969) thin film phenomena; Robert E. Krieger publishing company,
Huntington, New york p10-22.
8. Hideaki, A.et al (2008) preparation of thin film by sulfurization of stacked
metallic layers.thin solid film p 1457
9. Kai-feng Huang et al indium-nitrogen cooped zinc oxide thin film deposited by ultrasonic
spray pyrolysis. Journal of nanomaterial vol.2014. pp7.
10. Ludwig J. Gaukler, Dainius Perednis (2005) thin film deposition using spray pyrolysis.
Journal of electro ceramics. Vol. 14. Pp103-111
11. Jun-liag et al (2005) growth mechanism for N-doped Zn0 film grown by spray pyrolysis
method. Journal of inorganic material. 20(4). Pp959-964
12. D. Mohan et al (2009) deposition and characterization of ZnO thin film by modified
pulsed-spray pyrolysis. Semiconductor science and technology. Vol.24. pp8.
ISBN: 978-81-948129-1-3 Page 39

ALGINATE AS MEDICAMENT IN PHARMACEUTICS
Ashwini Ravi,* Hemapriya J and Vijayanand S
1. Department of Biotechnology, Dwaraka Doss Goverdhan Doss Vaishnav College

(Autonomous), Arumbakkam Chennai, Tamilnadu, 600 106.
2. Department of Microbiology, DKM College for Women, Sainathapuram, Vellore,

Tamilnadu, 632 001.
3. Bioresource technology lab, Department of Biotechnology, Thiruvalluvar University,

Serkkadu, Vellore, Tamilnadu, 632 115.
*Corresponding Author: ashwinir@dgvaishnavcollege.edu.in
ABSTRACT:
Alginate is a polysaccharide that is wide spread in brown sea weeds and some bacteria. They
were found to be present in the cell walls of brown algae and produced as capsulated
polysaccharides by earth bacterial species. The alginates also exist as salt mixtures of different
cations in sea water. The term alginate is commonly called as alginic acid but it is also used to
refer all its derivatives. It is a linear polymer consisting of two uronic acid units called as D –
mannuronic acid and L glucuronic acid. These two units are linked by β(1→4) and α(1→4)
glycosidic bonds. The surplus availability of alginates in environment and it’s not complicated
multi stage procedure of extraction makes it an effective candidate for its use in biomedical
applications. Despite easy extraction process, they have other advantages of low toxicity,
biocompatibility, mild gelating agent and can be produced at low cost. By varying the
composition of its monomeric units, the hydrogels formed by the alginates can be manipulated.
This property of alginates makes it to be used in cell transplantations and wound dressings. The
similarity in structure of alginates to the extracellular matrices of living tissues finds its
application in wound healing, delivery of bioactive drugs, chemical compounds and proteins. In
wound dressing, they provide moist microenvironment, prevent bacterial infection and promote
wound healing at the site of infection. In addition to these, they have also been used in cell
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cultures and tissue regeneration. In the present book chapter, the potential biomedical
applications of alginates in pharmaceutical industry will be discussed in detail.
KEYWORDS: Alginates, Brown algae, Drug delivery, Wound dressing, Tissue engineering.
3.1. INTRODUCTION:
Alginate is a polysaccharide extracted from brown seaweeds of family Phaeophyceaea and
was found to have β (1 → 4) mannuronic acid and α (1 → 4) guluronic acid unit [1]. It is located
at the intracellular matrix of sea weeds as gel containing sodium, calcium, magnesium, strontium
and barium ions [2]. They were found to give the sea weeds mechanical strength and flexibility
[3]. Later in the year 1964, alginate was discovered to be produced by bacteria such as
Azotobacter vinolandii, Azotobacter crococcum and Pseudomonas sp. [4 - 7]. The ability of
alginate to retain water, its gelling and stabilizing properties makes them to be widely used in
food industries and also in medicine.
Though alginates have no nutritional value their important properties such as viscosity
enhancement, gelling ability and stabilization ability makes them to be used to improve, modify
and stabilize the texture of some foods. They have been used in several food products such as ice
cream, sauces, Ketchup, Mayonnaise, frozen food, pastry fillings, puddings, pie fillings etc. In
the field of medicine, they have been used as disintegrating agent, thickening agent, stabilizing
agent and antacid stomach protector in tablets, suspensions, emulsions and in capsules [8]. In
modern medicine, they have been used as alginate matrices, microspheres and
microencapsulation for various processes such as drug delivery, protein delivery, tissue
regeneration, tissue engineering, wound healing, wound dressings etc. This chapter deals with
the application of alginate in the field of medicine.
3.2. PROPERTIES OF ALGINATES:

The main properties of alginates is that their ability to form gels. This is due to their ion
binding ability and sensitivity to various environmental factors such as temperature and pH.
These physical properties of alginates aids in its application in both food and medicinal industry.
Their important physical properties are discussed below:
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3.3. 1. SOLUBILITY:
The solubility of alginates differs with the type of alginate. For example, Calcium alginate is
insoluble in water and ether. Whereas it is slightly soluble in ethanol, completely soluble in
sodium polyphosphate, sodium carbonates and chemical substances that combines with calcium
[9]. Another form of alginate i.e. the sodium alginate dissolves in water and is insoluble in
ethanol and ether [10]. Other salts of alginate such as potassium and ammonium alginate are
soluble in both hot and cold water and magnesium salt of alginate is insoluble in water [11].
The solubility of alginates despite their different forms is dependent on three factors such as
pH, ionic strength of the solution and the gelling agents. The solubility of alginates based on pH
will be discussed later in the chapter, the ionic strength and gelling agents will be discussed in
brief. Alginates were found to be affected by the mixed solvents due to their dielectric constants
[12]. For instance, the mixed solvents of calcium and magnesium alginates have different
dielectric constants thereby affecting the activity coefficients of ions which affect their solubility
[13]. The solubility of alginates is also affected at higher ionic concentrations of 0.2 – 0.3 M due
to the altering water structures because of the cations and anions present in them which is
explained by thermodynamic function [14]. The solubility of alginate can be altered and can be
precipitated by salting out effects at a minimal concentration of 0.1 M salts. This salting out
ability of alginates is found to be used in separating polysaccharide mixture by precipitation [15,
16].
As far as its solubility is concerned in terms of gelling agents, alginates readily forms gels
with divalent cations such as Calcium. The gel network is formed by the intermolecular cross
liking between the carboxyl groups and divalent cations. This cross linking forms the basis of gel
formation and this phenomenon does not depends on temperature [3, 17]. At a concentration of
3mM above, ca2+ were found to precipitate all the alginates present in the solution and this
process is affected based on the amount of alginates and also its hydration. The hydration of
alginates, the rate of diffusion is even more affected than the individual alginate particle which
makes the process even slower [16].
3.3.2. ADHESIVITY:
Biodhesion is defined as the interfacial molecular attractive forces with biological substrate
and natural or synthetic polymer in order to adhere to the biological surface over a long period of
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time [18]. The process of bioadhesion involves several complex processes such as wetting,
adsorption, chemical adsorption etc. and is used for polymeric drug delivery system [19]. They
have been used in drug delivery for nasal ailments and they have also been proposed to be used
in gastrointestinal, ocular, buccal, vaginal and rectal ailments [18, 20].
Polyanionic polymers were found to be more efficient bioadhesives than the polycationic
and non – polymeric bioadhesives. Since alginates are anionic biopolymers with carboxyl groups
they are an efficient candidate to be used as bioadhesives. Alginates were proved to have more
mucoadhesive strength than other polymers such as chitosan, polystyrene, carboxy methyl
cellulose, polylactic acid etc [21]. As the research on alginates as bioadhesives continued, they
were crosslinked with several other biological materials in order to improve their function. The
alginates were crosslinked or modified with biomaterials such as catechol, gelatin, adhesion
ligand containing aminoacids, nanofibrous scaffolds etc. The methods in which they are bound
with the biomaterial has also been explored like chemical crosslinking, photopolymerization etc
[22 -25]. In order to study the effect of bonding with the biomaterial and to detect the extent of
its preparation methods like AFM (Atomic Force microscope) has also been employed [26].
3.3.3. BIOCOMPATIBILITY:
Alginates do not have any nutritional values but its physical properties of gel formation and
reacting with polymers of food components such as proteins under favourable conditions make
them to be used to maintain the structure of certain processed foods and pet foods. Therefore,
they have been used as thickeners, softeners and gelling agents in food industry [27]. As far as its
medicinal application is concerned, the use of alginate in the field of medicine first initiated with
transplantation of encapsulated pancreatic islets of Langerhans. They have been approved by the
FDA in the group of compounds that are used for oral applications. Alginates are non – toxic and
biodegradable when applied orally but when taken intravenously this was found to cause several
immunogenic ailments [28].
Since then several studies reports that alginates with M blocks were found to be
immunogenic whereas other reports show that G block showed much immunogenic activity than
the intermediate G blocks [29, 30]. Study by Tam et al. (2011) showed that in a alginate –
polycation microcapsules, alginate coating was found to have non – significant effects on the
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biocompatibility and physiochemical properties of the microcapsules. Other studies also show
that alginates have better biological properties to be used in medicine but still they are
inconsistent to be used while other studies shows no such activity [29, 32 - 35]. Despite their
contrary to be used in medicine, several works on use of alginate in medicinal field is still being
explored [29]. Houghton et al. (2015) showed that alginate can be used as weight loss agent since
they give zero calories.
3.3.4. pH SENSITIVITY:
Alginates will be greatly affected by pH of the solution. The effect of pH is very essential
since it is important to be used in medicine. It is an essential stimulus to be considered when
using in human body [37]. At low pH, the solubility is affected. At pH of 4 – 5 and even lesser,
alginates will be precipitated. Precipitation is caused due to the decrease of pH below the pKa
level of alginates [3]. At alkaline pH, of pH 10, alginates were found to swell and cause firm
gels than that of pH 1 [38]. The sensitivity of alginate to various pH is given in table below:
Table: Sensitivity of alginate to variable pH
Alginate type Acidic pH Alkaline pH
Alginate Insoluble Soluble
Sodium Alginate Insoluble Soluble
Potassium Alginate Insoluble Soluble
Calcium Alginate Insoluble Insoluble
Ammonium Alginate Insoluble Soluble
3.4. PHARMACEUTICAL APPLICATIONS OF ALGINATES:

As discussed earlier, the medicinal application of alginates includes its use in drug delivery,
protein delivery, cell culture, tissue regeneration, tissue engineering and in wound healing. These
applications of alginates have been discussed in detail below.
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3.4.1. DRUG DELIVERY:

Alginates, a natural polysaccharide is an important noticeable candidates used in the field of
drug delivery because of its biocompatabiltiy and ease in gelation these natural polymers are
used widely in medicine. This bioactive polymer has gained the attention of the scientific
community due to its tendency formulating controlled drug delivery system, where the release of
the drug can be controlled. According to [39] alginate combined with calcium acts as a better
electrode for delivering medication through Iontophoresis, the method where drugs are delivered
transcutanoeusly through skin. Alginates effectively transfers lidocaine using excised rat skin by
square wave altering current. These polymers are also used in the form of hydrogels. These
hydrogels are three dimensional networks composed of polymers in an interlinked manner.
Alginates are preferred as the best matrix for embedding islet cells in type I diabetes in rat
insulinoma cell lines. These polymers are usually combined with a retardant agent (Eg.Na 2PHO4)
which plays an important role in few physiochemical properties of the polymers like stiffness,
gelling time, porosity etc., [40].
Alginate when cross linked with chitosan was found to have 80% of drug releasing ability
in small intestine or colon. These dual cross linked beads are incubated in gastrointestinal tract
condition and found to release higher amount of BSA in SCF (Simulated colonic fluid) when
compared with Simulated Intestinal Fluid (SIF) and Simulated Gastric Fluids (SGF) [41]. Xing et
al. (2003) states that alginate when associated with calcium plays vital role in releasing drug in
specific site than being release in the physiological environment of the stomach. Alginate
together with sodium has been used in the formation of floating beads using freeze- drying
technique which has prolonged gastric residence time of more than 5 hours [43]. Ion- activated in
situ gel forming ophthalmic solution has been made using gatifloxacin, a broad spectrum
antibacterial agent. These include the formulation of alginate and HPMCE50Lv (Viscosity
enhancing agent) that forms drops on cul-de-sac upon instillation. These mixture exhibits
effective drug releasing tendency and high pre-corneal residence time of over 8h [44].
Cohen et al. (1997) also supports that aqueous solution of alginate with high G contents
such as Kelton LV acts as an effective polymer in the formation of ophthalmic solution which
delivers pilocarpine. Calcium-Alginate-NOOC beads were found to be better system for delivery
of protein drugs developed in aqueous medium at pH 7.4 [46]. Recent study revealed that
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alginate combined with graphene oxide exhibits high colon targeted drug releasing behavior.
These complexes also possess anti-tumour inducing ability in mice treated with GO-ALG/5-FU.
Thus they form a novel compound in treating colon cancer liver metastasis [47]. Nano alginate
particles when associated with Glycyrrhetinic acid, liver targeted molecules exhibits higher drug
delivering ability and distinct killing effect on hepatocellular carcinoma cells [48]. Ionotrophic
gelation of Graphane and alginate with Ca2+ forms composite beads which show dual response
to both pH and temperature with better cytocompatiability on HeLa cancer cell lines [49].
Alginate polymers are used in the formation of magnetic polymeric particles in drug delivery to
transcleral region of the eye [50].
Alginates have also been used in the major challenging fields like retroviral drug delivery.
Amide functionalized nanoparticles are prepared using emulsion solvent evaporation method.
The beads were found to be biocompatible and also deliver the drug efficiently in the target zone.
These made them as a promising carrier for delivery of retroviral drugs like zidovudine [51].
Alginate coated arabic gum and ghatti gum were microcapsulated using ionic gelation technique.
These microcapsules are used for intragastric delivery of flurbiprofen [52]. Drug delivery vehicle
has been made by fabricating polyurethane, alginate and chitosan for delivering insulin [53].
3.4.2. PROTEIN DELIVERY:

Alginate grafted with L-Arginine was found to be a suitable carrier for site specific protein
delivery. This combination possesses stability in varying pH and exhibits thermal stability. These
grafted polymers were found to be stable in dual gastric intestinal tract conditions (acidic and
alkaline) [54]. Alginate polymer as a vector plays an essential role in delivering drugs. Alginate
was found to be an effective carrier associated with eggshell derived calcium deficient
hydroxyapatite. This combination was found to be an ideal delivery agent by releasing 65% of
model protein (BSA) effectively [55]. Study by Wang et al. (2018) reveals that immune
protection against Streptococcus iniae in cat fishes were attained by vaccinating fishes with
alginate/chitosan encapsulated recombinant protein serine-rich repeat.
In vitro and in vivo study proves the stability and safety of microspheres. A study proposed by
George et al. (2007) shows that alginate together with guar gum was made into microspheres
using freeze dried method. These capsules were found to deliver about 20% and 90% of modal
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protein BSA at pH 1.2 and pH 7.4 respectively. Alginates have been used as a fabricating
material in preparing BSA loaded chitosan mediated nanoparticles for colon specific delivery of
proteins [58]. Alginate and aminated chitosan micro beads were found to be suitable protein
delivery system for intestinal and colon tracts [59]. Sodium alginate has been used as an effective
therapeutic protein delivering agent to wounds [60]. Zinc cross linked microparticles were
prepared using one step – spray drying process for effective controlled pulmonary delivery of
proteins [61]. Zein alginate based drug delivery system were found to withstand harsh
gastrointestinal condition and thus it act as an effective vector for delivering therapeutical
proteins [62]. Iron cross linked alginates- carboxy methyl cellulose beads also possess
controlled release of proteins in various simulated gastrointestinal conditions [63].
3.4.3. CELL CULTURE:

Cell culture has been developed for the artificial development of cells or tissue in in vitro
condition to be used in stem cell therapy or tissue replacement therapy. It is a complex process
for which factors such as strict environment, pH, temperature, oxygen and carbon dioxide levels
has to be maintained. Apart from these factors, the criteria determining the cell culture is its
matrix. The matrix is a medium which recreates the microenvironment required for the cells to
multiply and survive. In in vivo conditions, there are several mechanisms, supportive cells, and
extracellular matrix which consists of several proteins such as collagens, elastin etc. will be
present thereby enabling the division of new cells and keep them intact [64, 65]. Whereas, the
production and recreation of such microenvironment is a challenging process when the cells are
to be culture in vitro. To recreate that microenvironment, several biopolymers with collagen,
elastin, fibrinogen, hyaluronic acid etc. are prepared. Besides these, synthetic polymers such as
polyethylene glycol, acetylated hydrogels are also been employed for this purpose [66, 67].
Alginate hydrogels are formed by the ionic crosslinking of alginates with Calcium ions
[68]. These hydrogels was found to have high applicability in cell immobilization since the
production of alginate gels is easy and controlled. Alginate gels of different elasticity, softness
can be prepared and their ability to reverse the gelation found to be a pleasing character of
alginate to be used in cell culture, cell trapping and immobilization [69, 70, 71]. Alginates have
been used in stem cell culture to produce the stem cell niche essential for the cells to adhere and
grow. Silk – alginate based hydrogel was found to increase the elasticity and promoted the
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adherence of mouse embryonic stem cells and cell survival during transplantation.it also had
another advantage of adjusting the stiffness easily which is hard to be accompanied in other
matrices [72]. Calcium alginate gels were also found to increase the adherence and viability of
Human Mesenchymal stem cells (MSC) for the period of 6 weeks within the gels along with
quantifiable concentrations of human VEGF and bFGF making them an efficient candidate to be
used in stem cell culture [73].
Since 1940, cell culture is established in two dimensional systems that used glass or
plastic surfaces for the culture. Now, with the increase in medical facilities and new
technological developments, the cell culturing has been taken to the next level called as 3D cell
culture. Growing cells in 3D has advantages of developing and testing relevant drugs, culturing
multi cellular tissue and controlled physiological environment [74]. Alginates have been used in
the three dimensional cell culture and RGD – alginate was found to aid in the growth of Retinal
pigmented Epithelium [RPE]. RGD – alginate concentrations of 0.5 and 1 % were used to study
its efficacy in 3D culture of RPE cells and has been found that 0.5% RGD – alginate can
effectively derive, transport and transplant neural retina and RPE. It was also found to enhance
other pigment formation, neural and other epithelial tissue generation [75]. Alginates were also
found to maintain the viability of adenoma cells for a period of four months when they have been
cultured with human pituitary adenoma cells [76]. Chen et al (2015) explained the modification
of alginate microcapsules surfaces with Poly L Lysine graft Polyethylene glycol (PLL – g -PEG)
to generate an immunoisolation barrier for the culture of mouse induced pleuripotent stem cells.
This microcapsule created a microarray well that enabled cell survival and proliferation to form
single cell aggregates. Apart from their use in cultivating cells for transplantation in 3D culture
they have also been used for the invasion of anticancer drugs and its response for
chondrasarcomas in three cell lines CH2879, JJ012 and SW1353 [78]. In order to study the
effective formation of alginates and to produce these gels affectively an analysis method called
as Electro wetting on Dielectric (EWOD) Digital microfluids [DMF] has been demonstrated by
George and Moon (2015). They have explained the detail process of forming alginate gels with
calcium, its processes to remove the unreacted calcium from alginate, to purify, use and to
analyse the formed gel with EWOD DMF. This process enables the user to alter concentrations
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of alginate or calcium based on the gel formed during the analysis which helps to produce more
efficient gels for different processes.
Alginates have been extensively used in chondrocyte production. During the chondral
lesions, the chondrocytes cannot migrate due to the absence of pre – existing matrix. In order to
replace those matrices, alginate hydrogels have been used [80]. Alginates were found to enhance
proliferation, growth of cells and accumulation of agreecan and type II collagen around the cells
[81]. Chondrocytes were also found to be produced in alginate beads [82]. It was also found to
improve Human osteoblast cell clusters which can be used for correcting skeletal defects [83].
3.4.4. TISSUE REGENERATION:

Tissue engineering is a process that involves multidisciplinary principles such as
material science, medicine and life science in order to develop better tissue or organ of biological
structure and function. For this, a scaffold material is required to mimic the biological
environment for the tissue to multiple, grow, survive and transplant. An ideal scaffolding
material should have properties such as biocompatibility, suitable structure, mechanical strength,
degradation, ability to support cell adherence and also to allow retention of metabolic functions
[84, 85, 86]. Alginates have been used as scaffolds in tissue regeneration and also for delivery of
drugs, growth factors and therapeutically useful cells [87].
Alginates have been used to regenerate Schwann cells which are used to treat peripheral
nervous disorders. Scaffolds were prepared with fibronectin and Schwann cells by transforming
them into alginate matrix. Some of them were transplanted to PHB conduitand those cells were
found to be remained alive throughout the study. It has been proved that Schwann cells can
regenerate better in the presence of fibronectin and alginate than those without alginate [88]. The
two bone morphogenetic proteins rhBMP – 2 and rhBMP – 4 scaffolded with alginate gel was
delivered to femoral condyle of rabbit knees with osteochondral defects. Upon injection the
alginate created gel and helped the proteins to localize at the injection site. With the evaluation
of its effects after three months alginates were found to help the localization of proteins and the
protein rhBMP – 2 showed better regeneration than the protein rhBMP – 4 [89].
Alginates have been used in combination with bioglass scaffold to enhance the
periodontal regeneration. This composite were found to reduce swelling, degradation and
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enhance biomineralization and protein adsorption. Further they also helped in proliferation,
differentiation and viability of periodontal ligament fibroblast and osteocarma cells. They also
enhanced the activity of alkaline phosphatase activity of hPDLF cells in the composite scaffold
[90]. It has also been found that alginate microbeads incorporated with adipose derived stem
cells (ASC) have better delivering of viable cells facilitating regeneration [91]. Schizzi et al.
(2016) proposed a work where polymethacrylate was used with alginate for better performance
in tissue regeneration. This complex scaffold was found to increase the mechanical strength and
also provided suitable microenvironment for tissue regeneration. Wang et al. (2016) proposed
the preparation of scaffold using silk fibroin and sodium alginate. This scaffold was found to
have better porosity and structure for the utilization of soft tissue engineering. Similar work was
performed by Perteghella et al. (2017) where alginate was used with silk for growth and delivery
of Mesenchymal stem cells (MSC). Studies showed that they possess a suitable environment for
multiplication, differentiation and proliferation of Mesenchymal stem cells.
Alginates have been combined with polycaprolactone, fish collagen and phlorotannin for
the regeneration of hard tissues. This combination enhanced the proliferation of cells than the
collagen/alginate complex. They were also found to have higher calcium deposition and
osteogenesis than the control [95]. Alginate gels encapsulated with chondrocytes and chodregnic
growth media were found to hold great potential in cartilage regeneration [96]. Apart from these,
alginates have been made scaffold with several compounds such as collagen, core/shell struts,
Hydroxyapatite, chitosan and polymethacrylate [97 - 101]. A study on purification of alginates
and its use in regeneration showed that the immunogenic effects of alginates were found to
minimize than using the impure alginates [102].
3.4.5. WOUND HEALING AND DRESSING:

Gels have been used in wound healing for a long period of time since they prevent
the wound healing site from dehydration. Alginates were found to cure wounds except full
thickness wounds without occlusions since they provoke immune response in such kind of
wounds [103 - 105]. In another study, calcium alginates were found to provoke certain cellular
aspects of normal wound healing [106]. Qin (2003) showed that high M alginate and high G
calcium alginate were found to have high absorbing activity into the fibres which facilitate
wound healing. Alginate dressings were found to be used in healing of diabetic foot ulcers [108].
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Calcium alginate/Polyvinylalcohol blended nanofibre matrix were efficiently used as

wound dressing and found to provoke growth of epithelial cells without any harmful effects
[109]. Apart from these, alginates have been combined with silver, chitosan and antimicrobial
agent like simvastatin to be used in wound healing and dressing [110 - 116].
3.5. CONCLUSION:
Alginate was found to have better medicinal applications and has better biocompatibility.
The gelling ability of alginates by the formation of ionic cross linking makes them to be used in
several ailments such as wound healing, dressing, drug delivery, tissue regeneration etc. When
they are modified with further biocompatible molecules or biologically derived compounds their
immunogenic response may also be reduced. However, in the field of medicine, alginates form a
promising compound to be used in the above mentioned methods to treat various ailments.
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106. Doyle, J.W., Roth, T.P., Smith, R.M., Li, Y.Q., and Dunn, R.M., Effect of calcium alginate
on cellular wound healing processes modeled in vitro. J. Biomed. Mater. Res., 32(4), 561-
568, 1996.
107. Qin, Y., Absorption Characteristics of Alginate Wound Dressings. J. Appl. Polym. Sci.,
91(2), 953–957, 2004.
108. Dumville, J.C., O’Meara, S., Deshpande, S., and Speak, K., Alginate dressings for healing
diabetic foot ulcers (Review). Cochrane Database Syst. Rev., 2, CD009110, 2012, DOI:
10.1002/14651858.CD009110.pub2.
109. Tarun, K., and Gobi, N., Calcium alginate/PVA blended nanofibre matrix for wound
dressing. IJFTR, 37, 127 – 132, 2012.
110. Opasanon, S., Muangman, P., and Namviriyachote, N., Clinical effectiveness of alginate
silver dressing in outpatient management of partial-thickness burns. Int. Wound J., 7(6),
467–471, 2010.
111. Devi, M.P., Sekar, M., Chamundeswari, M., Moorthy, A., Krithiga, G., Murugan, N.S.,
and Sastry, T.P., A novel wound dressing material— fibrin–chitosan–sodium alginate
composite sheet. Bull. Mater. Sci., 35(7), 1157–1163, 2012.
112. Percival, S.L., and McCarty, S.M., Silver and Alginates: Role in Wound Healing and
Biofilm Control. Adv. Wound Care, 4(7), 407 – 414, 2015.
113. Yu, W., Jiang, Y.Y., Sun, T.W., Qi, C., Zhao, H., Chen, F., Shi, Z., Zhu, Y.J., Chen, D.,
and He, Y., Design of a novel wound dressing consisting of alginate hydrogel and
simvastatin-incorporated mesoporous hydroxyapatite microspheres for cutaneous wound
healing. RSC Adv., 6(106), 104375–104387, 2016.
114. Dai, M., Zheng, X.L., Xu, X., Kong, X.Y., Li, X.Y., Guo, G., Luo, F., Zhao, X., Wei, Y.Q.,
and Qian, Z., Chitosan-Alginate Sponge: Preparation and Application in Curcumin
Delivery for DermalWound Healing in Rat. J. Biomed. Biotechnol., 2009, Article ID
595126, 1 – 8, 2015, doi:10.1155/2009/595126
115. Trial, C., Darbas, H., Lavigne, J.P., Sotto, A., Simoneau, G., Tillet, Y., and Téot, L.,
Assessment of the antimicrobial effectiveness of a new silver alginate wound dressing: a
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116. Straccia, M.C., D’Ayala, G.G., Romano, I., Oliva, A., and Laurienzo, P., Alginate
Hydrogels Coated with Chitosan for Wound Dressing. Mar. Drugs, 13(5), 2890-2908,
2015, doi:10.3390/md13052890
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HARMFUL BIOLOGICAL EFFECTS OF PLASTICS
Yusuf Sarkingobir, Ummu Tukur
Department of Biology Shehu Shagari College of Education Sokoto, Nigeria
Email: superoxidedismutase594@gamail.com
ABSTRACT
The aim of this paper is to discuss harmful Biological Effects of plastics. Plastic materials are
ubiquitous and everywhere you go in our environment. This is because of our increased demand
and poor management of plastics. Therein, these materials find their way into diverse biological
systems on land and water, and lead to public health problems. Plastics become fragmented or
degraded due to mechanical or chemical actions respectively to yield smaller particles, which in
turn enters the body through diverse routes more especially via the mouth. It can also adsorbed
or absorbed other chemicals (including toxins).In the body ,plastics can be retained or
internalized in the gastrointestinal tract (or other cells or tissues) or translocated to various places
(organs and tissues); biomagnification can also occur. Albeit little is known about the exact
hazards of plastics on biological systems (more especially in humans), some studies were
documented. Plastics can easily leach toxic chemicals to its contacts (biological system or
anything nearby such as food) as a result of heat or ultraviolet rays. Plastics additives or
monomers are associated or implicated with many adverse effects. Parable, the plastics (or
additives) are associated with increased risk of impaired brain and neurological functions,
cancer, chemotherapy delay, obesity, early puberty, drug resistance, adult-onset diabetes,
chromosomal and reproductive system abnormalities, cardiovascular damage among others.
When plastics are burnt, hazardous substances are released such as dioxin; a carcinogenic,
hormone disruptor and persistent chemical. Mercury, furan, formaldehyde, hydrochloric acid and
relations are also released on burning plastics; especially the home practice. Burning of plastics
increases the risk of respiratory problems like asthma. It causes rashes on the body, nausea,
headache, damages nervous system, and kidney, liver and reproductive system. Styrene, a
component of polystyrene present in our air and can be inhaled. It is associated with increased
cancer risk, it has reproductive effects, and affect central nervous system(CNS) to cause
(depression, fatigue, weakness, hearing loss etc). Ranges of exposure to vinyl chloride can cause
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effects on nervous system, liver damage, and cancer. Moreover, Plastic production and recycling
consume much energy, in turn releasing greenhouse gases, and ultimately stimulating climate
change. Thus, it is time now to put a stop or strict measures to curtail the escalating trend of
plastics dangers. Recycling, reducing, refusing, improvisation, banning and taxation, are some of
the clues to reduce the menace of plastics .
KEYWORDS: Plastics, biological system, dioxin,pthalates
1. INTRODUCTION
Waste is presently a global, regional, and local issue affecting public health and other
organisms. Therein, plastic is one of the leading products that contribute hugely to waste
generation and resultant consequences. In one hand, plastic has a diverse uses to make life
easier; and in the other hand, plastic lifecycle is characterized with disadvantages to biological
systems on earth .In Nigeria, more than 23,400,00 tones entered our environment from 1996-
2014.Babayemi et al., (2018) admits plastics as a pressing threat to public health in Nigeria
which need urgent attention. Kaoje et al., (2015) and Ibrahim et al., (2019), Sarkingobir et al.,
(2019) reported plastics as the most predominant waste in Sokoto environment. However, a
literature review performed revealed that there is scarcity of concise research that described
plastics use and its consequences; both at international, national, and local levels. Plastic is a
word that originated from Greek, meaning “pliable and easily shaped.” Recently it became a
name for a certain materials called polymers. The word polymer means “of many units,” and
polymers are made of long chains of molecules. Thus, plastics are pliable polymers that are
easily shaped. This gave them applications in almost every aspects of our everyday life
(Sarkingobir et al., 2020). Thus, this paper sought to make a laconic review, of harmful
biological effects of plastics, so as to create awareness among the stakeholders and stir positive
decision makings.
2. COMMON PLASTICS AND ITEM CONSUMER IDENTIFICATION CODES

The society of the plastics industry (SPI) formed a classification system for guiding
consumers and recyclers to identify different types of plastics. Each specific plastic is given a
number code, which is usually shown on the plastics product. But, many consumers are unaware
of these codes or even if they see them they cannot fully identify their meanings. There is need
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for creating more awareness among the consumers, so that they can be guided to form
meaningful choice when using plastic products. The table below shows the SPI posted on plastic
packaging(Azoulay et al., 2019)
S/N PLASTIC GENERAL PROPERTIES ITEMS WHERE IT CAN COMMENT

TYPE (CODE) BE FOUND
1. polyethylene Good and moisture burner Mineral/ water/ food trays, Sometimes absorbs odours,
terephthalate properties clear, hard, tough, beer bottles, soft drink flavours from food or
microwave transparency, /water bottles, fibre of cloth anything stored in them
solvent resistant and carpets mouth wash
bottle
2. High Density Excellent moisture burner Detergent, bleach, bottles, Never safe to re-use for
polyethylene properties, chemical food boxes carbonated food or drink
resistance, hard semi – drinks bottle, buckets, rigid
flexible permeable to gas piles, toys, garden furniture,
HDPE films crinkle to touch bins
3. Polyvinyl Excellent transparency hard, Carpet, credit cards, It is harmful if ingested, it
chloride PVC rigid (if plasticized) good windows, frames pipes, should not be in contact
chemical resistance, good fittings, wire, cable sheath, with food or drink
weathering ability, stable synthetic leather tiles, rest sometimes recyclable often
electrical properties, low gas mats, cosmetics food contain lead, phthalates,
permeability packaging dolls, food releases disorient after burn
package
4. Low density Tough, flexible waxy Films, fertilizer bags, Acclaimed to be healthy
polyethylene surface, soft, very transparent bubble wrap, irrigation sometimes recyclable
LDPL stable electrical properties, pipes, clothes, wire, cables,
good moisture burner some bottle tops.
5. Polypropylene Very chemical resistant, high Most bottle tops, syrup Occasionally recyclable
PP melting point hard, flexible bottles, yoghurt bottles,
wave surface, translucent crates, biscuit wrappers, car
pets straws, fabrics.
6. Polystyrene PS Clear to opaque, glassy Egg boxes, yoghurt Commonly recycle, but
(Styrofoam) surface, rigid or foamed, hard containers container, video difficult suspected
brittle, not resistant to fats castle, seed trays, low cost carcinogen and neurotoxin.
and solvents, high clarity. toys, hangers, disposable
cutlery meat fish packages.
7. Other Refers to diverse polymers Nylon (PA) polycarbonate Difficult to recycle PC is
other than the ones stated (PC) acrylonitrile butadiene used in compact disc, baby
above styrene (AB) bottles, medical containers
contains bisphenol
A(BPA), a hormone
disrupter
Adapted from Harrison and Beedard (n.d) Eco-Healthy Child Care(2010), Katz(1998),
Babayemi et al., (2018), , Valentine(2018). Azoulay et al. (2019),
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3. THE DANGERS OF BURNING PLASTICS AT HOME AND RELATIONS

Burning of plastics increase the risks of asthma, nausea, heart disease, emphysema, rashes and
headache. It can also harm the nervous system, liver, and reproductive system. In burning,
pollutants are released, which in turn travel through the air to affect proxy or neighbours some of
these pollutants are mercury, dioxins, furans, and polychlorinated biphenyls (PCBs). They can
persist for long time in the environment in our water, food, or can bio-accumulate in the
biological systems (e.g of wildlife, fishes, meat and dairy) (Lake County Environmental Health,
2013).
POLLUTANTS RELEASED DURING BURNING
Dioxins
 Released by items containing trace chlorines.
 They are persistent in water, vegetation and food.
 Bio accumulate
 Immune suppression, hormone disruption, cancer particulate matter.
 Can enter lungs.
 Can cause heart attack or irregularities
 Reduces visibility
Polycyclic Aromatic Hydrocarbons (PAHs)
Some of them are cancer causing agents.
HCL
Produce on burning PVCs

Cause dermatitis, skin burns, laryngitis, trachealis, hoariness, choking, bronchitis, pulmonary
oedema, nausea, vomiting, diarrhoea, dehydration, convulsions, chills, shock, stupor, circulation,
collapse, visual damage.
Heavy Metals
Such as lead, mercury, cadmium, chromium, Arsenic Carbon monoxide
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Formaldehyde
Can cause cancer, cause watery eye, difficulty in breathing (Women in Europe for a Common
Future, n.d; Eco-Healthy Child care, 2010; Lead County Environmental Health, 2010;
Saskatchewan Ministry of Environment, 2012; Verma et al.,2016; Azoulay et al., 2019)
4. IMPACTS OF PLASTIC ON CLIMATE CHANGE

The present greenhouse gases (GHGS) emissions from plastic life cycle threaten the global
community to keep the global temperature rise below 1.5 0c. If the current emission and
production continues, by 2030, the emission from plastics will reach 1.34 gigatonnes. The
emission of gases that are pro-climate change is involved in even stage of the life cycle of
plastics, beginning from fossil fuel retraction and transport, plastic refining and manufacture and
transport, plastic refining and manufacture, management of plastic waste, plastics impacts on
oceans, water way and landscape. In retraction and transport section, fossil fuel is harvested and
thus produces significant greenhouse gases. It includes direct emissions, such as of melting,
flaring, fuel combustion and energy consumption in drilling of oil and gas, emissions due to
deforestation (European Commision, 2011).
In plastic refining and manufacture emissions are made. This sector accounts for one of
the most significant GHGS emissions and is faster growing. It requires much energy in cracking
of alkanes, polymerization, plasticization and quasi. Plastic waste management sector also
releases emission through land drilling, recycling or incineration. The least emission comes from
land drill, then recycling and lastly incineration. Plastic in the environment is also a threat,
therein, it contributes to degradation of the environment. Studies at an early stage shows that
plastics on coastlines, river banks, landscapes, continue to releases methane and other GHGS in a
cascade manner (that is one release, triggers further releases), albeit, the emission from the inner
and underground are yet to be qualified with full precision. Also, the micro plastics in water
reduces the ability of water (Oceans) to act as carbon sink (there by reducing climate change) by
reducing the ability of photo plantation and relations (Koushal et al, 2014; Azoulay et al., 2019;
Hamilton et al., 2019).
CHEMICALS RELEASED AT EXTRACTION AND TRANSPORT OF PLASTICS

Before plastic releases the consumers or environments severe human health effects are evident
in the extraction and transport storage. Thus, there is need to understand things from the onset.
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5. HARMFUL CHEMICALS ASSOCIATED WITH PLASTICS

There are diverse array of chemicals that are emitted during the lifecycle of plastics and the
present diverse hazards to biological system. For example, during interaction and transport,
heavy vehicles requires diesel and releases chemicals like BTEX and particulate matter, which
can stir respiratory problems like asthma, shortness of breath e.t.c. Other chemicals are Ozone,
Volatile organic compounds (VOCs), mixed oxides of nitrogen (NOx). Exposure to ozone in a
chronic fashion impair lungs function, in fact about, 1, 606 are associated with oil and gas
extraction, which 173 are associated with cancer, neurotoxicity, immune system effects, liver and
kidney problems, and reproductive problems (Science for Environment Policy, 2011; Koushal et
al., 2014; Azoulay et al., 2019).
6. CHEMICALS RELEASED DURING PLASTICS PRODUCTION
1,3 – butadiene
Is used as a monomer make rubber, plastic and relations. Short term exposure to this chemical
lead to headache, fatigue, low blood pressure, central nervous system damage, and
unconsciousness. Long-term exposure can cause cancer and chances of leukaemia.
Benzene
Is used as a solvent in making monomers during polymerization. Benzene is a bone marrow
poison. In short exposure it causes headache, tremors, drowsiness, dizziness, and high exposure
can cause death. I was shown to be associated with blood cancer.
Styrene
It can travel through the air to food and our body. Styrene exposure causes irritation of eyes,
skin, and nose. High exposure causes changes invasion, slow reaction times, cancer and problem
in maintaining balance (Azoulay et al, 2019).
Toluene
Toluene is used as solvent in production of plastics. Short-term exposure causes fatigue,
weakness, memory loss, nausea and loss of appetite. Long-term exposure causes irritation of the
eyes, or lungs, headache and dizziness.
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Ethane
It is used in production of ethane, during which S02, nitrogen oxides, VOCs, lead CO and O3
can be produced.
Propylene and propylene oxide

Propylene is used in the production of plastic fibers. Moderate exposure to propylene causes
dizziness, drowsiness, and unconsciousness. Propylene oxide is used in production of
polyurethane and polyethers. Propylene oxide is a classic carcinogen.
Polycyclic aromatic hydrocarbons (PAHS).

There are about 100 chemicals known as PAH. They are environmental toxicants and have
negative health impacts. PAHs such as anthracene, perantherene, and pyrene can cause
reproductive problems, tumours, low birth weight and birth defect is in mice. Anthracenes cause
headaches, nausea, loss of appetite, and inflammation of stomach and intestines. It also delays
reaction time, and weakness. (Saskachewan Ministry of Environmental, 2012; Azoulay et al.,
2019).
7. CHEMICALS RELEASED BY CONSUMER PLASTICS

Consumer plastics basically contain 3 things, viz, resins, fibers, and additives. The resins
(monomers) can contain remnants of un-polymerized units which can leach to food or other
materials that are in contact with the plastics product some these plastics even carcinogenic.
They include polyurethanes (flexible foam in furniture, bedding, carpet backing), PVC (pipes,
wire cable coatings, packaging), epoxy resins (coatings adhesives, composites like carbon fiber,
fiber glass) polystyrene (food packaging, CD cases, hard plastic in consumer products) (Sciences
for Environment Policy, 2011). In the other hand, a wider range of chemicals are added to
plastics to confer specific properties. Nearly, 93% of plastics is composed of monomers, while,
the 7% composed additive. The plastic with highest number of additives is PVC (SEP, 2011) are
of diverse functions. Softeners and plasticizers are added for flexibility, stabilizers and
antioxidants are for durability against heat or sunlight and flame retardants. The most said toxic
additives are flame retardants (polybrominated biphenyl’s(PBDE) polychlorinated biphenyl’s
(PCBs) phthalates, lead compoundsJoint (Women in Europe for a Common Future,n.d.; Women
ISBN: 978-81-948129-1-3 Page 69

in Europe, n.d.; Group of Experts on the Scientific Aspects of Marine Environmental Protection
,GESAMP; 2015; Laville, 2019; UNEP Prontiers, 2016; Romje, 2019; Rosane, 2019).
8. BIOCHEMISTRY OF PLASTICS IN BRIEF

Fragmentation - breaking down to smaller pieces through physical means. Degradation -
breaking down to smaller pieces through chemical means
Adsorption and absorption (Rist and hartmann, 2017; Giedre and Bethanie, 2018 ; Leslie, 2014).
Plastics ingested can bioaccumulate, or translocate (transported ) within the body, then parts of it
can be egested (Leslie, 2014; Schwabl, 2014). Biomagnification, increasing in concentration,
along the food chain at the higher level . Plastics interaction causes-oxidative stress,
inflammation, granulocytoma formation, lysosomal membrane destabilization ,neutrophil trap
release ,cytokine regulation, non-specific responses (internalization) of plastics ,lipid
peroxidation, necrosis, perturbation of membranes ,activation of detoxification pathways ,DNA
stand break(Brande – levindsen, n.d.; Gopal et al., 2014;Centre for the Study of the
Economies,2018; EarthDay Network,2018; Giofra, 2018; Environmental Investigation
Agency,2019; Food and Agriculture Organization of the United Nations,2019; Fox,2019;).
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Example of animals negatively impacted by plastic

Source: Machucha,2013; Leslie,2014
9. BIOCHEMICAL EFFECTS OF PLASTICS

At tissue-organ level, plastics destroy GIT, mucous production, disturb lipid and hepatic
metabolism, glycogen depletion, fatty acid vacoulation. At organismal level - causes false
satiation (loss of appetite), reduced fitness, and reduced survival, impaired respiration ,impaired
development and reproduction, increased mortality (Giedre and Bethanie, 2018; Rist and
Hartmann, 2017).
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10. CONCLUSION
Plastics have diverse array of harmful effects to biological systems.
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affect your health .Toxic Ameriac.www.theguardian.com
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on the environment : balancing the knowen and unknown .Goteborgs University .
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15. Hamilton, L., Feit, S., Kelso, M., Rubnigth, S.M., Moon, D., Morris, J., Labbe – Bellas, R.
(2019). Plastic and climate; the hidden cost of a plastic planet. www.c.cl.org/plastic and
climate.
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M.A.(2020). The dangers of plastics to public health: A review. Journal of Science and
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SURVEY ON WETLAND PLANTS OF PERIYAKULAM POND IN VALLIOOR,
TIRUNELVELI DISTRICT, SOUTH INDIA
J. Albino Wins, Selvanandhini.T and N.Nishanthi
Department of Botany, Holy Cross College (Autonomous), Nagercoil – 4
(Affiliated to Manonmaniam Sundaranar University, Abishekapatti, Tirunelveli District)
ABSTRACT:
The biodiversity survey on wetland plants in the village area envisaged exploration and
documentation of the plant diversity of Periyakulam pond in Tirunelveli district. The present
investigation explores the enumeration of plant species, floristic analysis of taxa. The study is the
result of intensive exploration carried out during the September2020-October 2020 (2months). In
the present study, a total number of 26 vascular plant species were recorded.
KEYWORDS:
Wetland plants, Periyakulam pond, Taxonomy, Vascular plants.
INTRODUCTION
Aquatic and wetland plants are mostly confined to the marshes and wetland habitats
(Cowardianetal., 1979). Although wetlands cover only six percent of the earth’s surface
(Mitschet al.,1993), they provide habitats for about 20 percent of the earth’s total biological
diversity (Gopal 1995 ; Les et al., 1995). Wetland supports plant species intermediate between
true aquatic and terrestrial habitats (Banerjee and Venu, 1994).Wetland ecosystems typically
show three characteristic ecological conditions, all of which are potential stressors for plant
survival and growth: periodic to continuous periodically anoxic (hydric soils); and hydro soils
with rhizospheres experiencing periods of low or no oxygen availability (Craft et al., 2005).
Freshwater ecosystems refer to ponds, lakes, springs, streams, rivers and wetlands. They are one
of the most productive ecosystems of the world and are rich storehouse of floral and faunal
diversity (Guliaet al., 2017). The first and foremost process in appreciating the biodiversity is the
taxonomic treatment of living organisms. Without appropriate knowledge of the exact species
composition, it will become very difficult to identify and implement conservation priorities for
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any ecosystem (Ragavan et al., 2016). Therefore, the present study was conducted to prepare an
inventory of floristic diversity of plants found in wet periphery of village ponds of Tirunelveli
District in Tamil Nadu.
METHODOLOGY
The present study was carried out in Periyakulam pond in Vallioor, Radhapuram Taluk in
the Tirunelveli district, Tamilnadu, India. The methodology used is observation method for the
determination of flora. Field data regarding their habit, habitat, phenology and flower colour
were noted. The survey involved several visits to the site for collection of samples and
photographs of the natural remnant vegetations and plantations.
All the specimens collected were identified with the help of recent literature by local
floras (Hooker 1872-1897; Gamble and Fischer, 1915-1935; Henry et al., 1989; Mathew, 1993).
The identification is authenticated by matched with type specimens available in the herbarium of
Botanical Survey of India, Southern Circle, TNAU Campus, Coimbatore, Tamilnadu. The plant
taxa were arranged to families according to APG IV (2016) system of classification. For all
documented species, the binomial and author citation were checked thoroughly with IPNI (2012)
and The Plant List Version 1 (2010) available online.
RESULTS AND DISCUSSION

In the present study, a total of 26 species of vascular plants were enumerated. Our results
coincide with earlier report made by various works in India, Nambineri wetland of
Gopalasamuthiram village, Tirunelveli district (Lakshmananand Ganthi, 2018).
Table :1 List of wetland angiosperms in Periyakulam ponds,Tirunelveli District,

Tamilnadu, South India
S.NO BOTANICAL NAME FAMLY TAMIL NAME HABIT CLASS

1 Lantana camara L. Verbenaceae Unnichedi Shrub Dicot
2 Ficus religiosa L. Moraceae araca-maram Tree Dicot
3 Cocos nucifera L. Arecaceae Tennai Tree Monocot
4 Azadirachta indica A.Juss Meliaceae Veppai Tree Dicot
5 Pistia stratiotes L. Araceae akasha-t-tamarai Herb Monocot
6 Ruellia tuberosa L. Acanthaceae Chetapatakaayala Herb Dicot
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7 Gomphrena serrata L. Amaranthaceae Herb Dicot

8 Mimosa pudica L. Leguminosae Thottarsinungi Herb Dicot
9 Pergularia daemia (Forss Apocynaceae Veliparutthi Climber Dicot
k.) Chiov.
10 Trichosanthes tricuspidat Cucurbitaceae kurattai Climber Dicot
a Lour.
11 Gloriosa superba L. Colchicaceae Chenkantal Climber Monocot
12 Tridax procumbens (L.) Asteraceae Vettukaaya poondu Herb Dicot
L.
13 Leucas aspera (Willd.) Lamiaceae Thumbai Herb Dicot
Link
14 Acalypha indica L. Euphorbiaceae Kuppaimeni Herb Dicot
15 Achyranthes aspera L. Amaranthaceae nay-urii Herb Dicot
16 Alternanthera sessilis (L.) Amaranthaceae Ponnanganni Herb Dicot
R.Br. ex DC.
17 Centella asiatica (L.) Apiaceae vallarai Herb Dicot
Urb.
18 Oxalis corniculata L. Oxalidaceae Paliakiri Herb Dicot
19 Ipomoea aquatica Forssk. Convolvulaceae Sarkaraivalli Herb Dicot
20 Cyanthillium cinereum (L Compositae Puvamkuruntal Herb Dicot
.) H.Rob.
21 Colocasia esculenta (L.) Araceae Cempu Herb Monocot
Schott
22 Nerium oleander L. Apocynaceae Arali Shrub Dicot
23 Datura metel L. Solanaceae Karu ūmattai herb Dicot
24 Crotalaria pallida Aiton Leguminosae Kilukiluppai Herb Dicot
25 Parthenium hysterophoru Compositae Shrub Dicot
s L.
26 Abutilon indicum (L.) Malvaceae Paniyaratutti Shrub Dicot
Sweet
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Out of the 26 species of wetland angiosperms, dicotyledones are represented by 22 plants

belonging 18 families, while monocotyledones contributed by 4 species belonging 3 families and
only one pteridophyte.
Distribution details of collected taxa in different plant groups were arranged based on APG
IV System of classification. Clades with maximum number of species includes Rosids with
8species belonging to 7 orders followed by Asterids with 11 species belonging to 6 orders,
monocots with 8 species belonging to 5orders, Superasterids with 3species belonging to 2 orders.
The details of clades were represented in Graph 1.
Graph 1. Distribution of species in clades/ grades as per APG IV system of Classification
Orders with maximum number of species includes Amaranhaceae with 3species and its
followed by Araceae, Compositae,Apocynaceae with 2 species,
Moraceae,Arecaceae,Meliaceae,Lamiaceae, convolulaceae, Oxalidaceae,Asterales with one
species (Graph 2).
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Graph 2: Distribution of species in dominant order as per APG IV system of Classification
Families with maximum number of species includes Amaranthaceae with2 spiecies, followed by
Apocynacae, Compositae ,Araceaewith 2 species .The 4 dominant families with the details of
number of species and genera are given in Graph 3.
Graph 3. Dominant plant families of wetland angiosperms
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REFERENCE
1. Adhikari S. Bhupendra & Mani M. Babu 2008, Floral diversity of Baanganga

wetland,Uttarakhand, India, Check list, vol. 4, no.3, pp. 279-290.
2. Bhagyaleena, P & Gopalan, R 2012a, Aquatic medicinal plants in ponds of
Palakkad,Kerala, India, Journal of Pharmacy and Biological Sciences, vol. 2, no. 3, pp.29-
35.
3. Craft CB, Malcom A, John W, Jeffrey M, 2005, Natural and Constructed Wetlands. In
Encyclopedia of Hyrological Sciences, pp. 1639-1655.
4. Chapman, A. P., B. W. Brook, T. H. Clutton-Brock, B. T. Grenfell and R. Frankham
(2001). Population viability analyses on a cycling population : a cautionary tale.
Biological Conservation, 97 : 61-69.
5. Deka, U & Sarada Kanta Sarma 2014, Present status of aquatic macrophytes of the
wetlands of Nalbari district of Assam, India, Pelagia Research Library, vol. 4, no.3, pp.
67-75.
6. Gopal B, Wetland and biodiversity: How to Kill Two Birds With One Stone ? In:
W.Giesen (Ed.). Wetlands Biodiversity and Development. Proceeding of Workshop of the
International Conference on Wetlands and Development held in Kuala Lumpur, Malaysia,
and 9-13 October 1995. Wetlands Internationals, Kuala Lumpur,1997, pp18-28.
7. Hooker, J.D. (1872-1897). Flora of British India, Ashford: Reeve and Company, Vol. 1-7,
5568pp.
8. Jain, A, Roshnibala, S, Kanjilal, PB, Singh, RS & Birkumar Singh, H 2007, Aquatic/
semi- aquatic plants used in herbal remedies in the wetlands of Manipur, Northeastern
India, Indian Journal of Traditional Knowledge, vol. 6, no. 2, pp. 346-351.
9. Kumar, V & Singh, PK 2012, Aquatic and semi- aquatic flora (Monocotyledons) of
Lalitpur district (U.P), India, Journal of Environmental Science, Toxicology and Food
Technology, vol.1, no.1, pp. 26-28.
10. Les DH, Schneider EL, In: Rudall PJ, Cribb PJ, Cutler DF, Humphries CJ, (Ed), 1995,The
Nymphaeles, Alismatidae, and the theory of an aquatic monocotyledon origin.
Monocotyledons: Systematics and Evolution, Royal Botanic Gardens, Kew, pp 23-24.
ISBN: 978-81-948129-1-3 Page 80

11. Lakshmanan R and Saravana Ganthi A, 2018, Phytodiversity studies of Nambineri

wetland of Gopalasamuthiram village, Tirunelveli district, Tamil Nadu, JMPS, Vol.6,
No.6, pp.106-115.
12. Prasad SN, Ramachandra TV, Ahalya N, Sengupta T, Kumar A, Tiwari AK, 2002,
Conservation of wetlands of India- a review. Trop. Ecol. Vol. 43,No.1, pp.173-186.
13. Rasingam, L, 2010, Aquatic and wetland plants of little Andaman Island, India, Journal of
Basic and Applied Biology, vol. 4, no.3, pp 52-59.
14. Sukumaran, S & Raj, ADS 2010, Medicinal plants of sacred groves in Kanyakumari
district Southern Western Ghats, Indian Journal of Traditional Knowledge, vol.9, no. 2,
pp. 294-299.
15. Swapna, MM, Prakashkumar, R, Anoop, KP, Manju, CN & Rajith, NP 2011, A review on
the medicinal and edible aspects of aquatic and wetland plants of India, Journal of
Medicinal Plants Research, vol. 5, no. 33, pp.7163-7176.
ISBN: 978-81-948129-1-3 Page 81

NANOPLASTICS: SMALL SCIENCE WITH BIGGER CONSEQUENCE

Yusuf Sarkingobir, Aminu Umar Imam, Kasimu Abubakar Shagari
1. Department of Biology, Shehu Shagari College of Education Sokoto, Nigeria
2. Department of Biochemistry, Sokoto state University, Sokoto, Sokoto state, Nigeria
3. Department of Integrated Science, Shehu Shagari College of Education Sokoto, Nigeria
Email: superoxidedismutase594@gamail.com
ABSTRACT
This paper is discussing on the consequences of nanoplastics on biological systems in a
laconic fashion. Life is likely unimaginable without plastics, because they are extensively useful
in all aspects of our lives. They are everywhere you. Nanoplastics are nano-size plastics with
promising features distinctively from other macromolecules. The tiny nature of nanoplastics is
responsible for their promising and effective applications. On the other hand, they undergo
certain processes and properties which make them consequentially harmful to biological systems.
They are easily inhaled, ingested or absorbed into the body. They are easily translocated within
the body and can even traverse the blood brain bilayer. They can cross most of the layers. They
can accumulate, magnify, retent, or be egested. In the body, they lead to destruction of lipid
bilayers, and causes inflammation of many tissues and organs, then oxidative stress. There are
other consequences of Nanoplastics. Therefore, diverse attempts are required to control the
situations.
KEYWORDS: lipid bilayer, Nanoplastics, inflammation, fragmentation, harmful effects,

inhalation, leaching, bioaccumulation
1.INTRODUCTION
Plastics are polymers with large molecular weight that can be moulded. The polymers in
plastics may be homopolymers (containing identical monomer units), and heteroplymers
(containing non-similar monomer units). Plastics also contain in addition to polymers other
ingredients such as fillers, plasticizers, stabilizers, colourants, etc (GESAMP, 2015). Presently,
commonly used plastics in our midst are: polyethylene ( in plastic bags, storage containers),
polypropylene (in rope, bottle cups, gear), polystyrene(cool boxes, containers), polyethylene
terepthalate (bottles, strapping) and others (GESAMP, 2015; Ibrahim et al., 2019; Sarkingobir et
al., 2019; Sarkingobir et al., 2020). Nanoplastics definition has been extensively debated by
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scientists and specialists. Nevertheless, nanoplastics are plastics of nano-scale or nano-size.

Thus, it is a parcel of microplastics (Rist and Hartmann, 2017; Stapleton, 2019). The smaller/
ultra size of Nanoplastics (NP) gave them large surface-to-volume-ratio, therefore becoming
very reactive, and more applicable and effective in many respects. It confers them with the
property of been very difficult to determine. It also makes them easily absorbable, translocable,
and ability to traverse layers of the body. Thus, concerns are raised. Common source of
contacting nanoplastics by humans or biological systems are textiles, tires, roads, paints, beers,
salts, honey, sugar, toothpaste, cosmetics, fertilizers, and detergents(Rist and hartmann, 2017;
Stapleton, 2019, Kogel et al., 2020; Prust et al., 2020). Others are exfoliators, shower gel.
Nanoplastics are hidden in many products around us nowadays. The Nanoplastics have an array
of applications, and in the other hand a wide array of harmful effects to the biological systems.
The smaller size makes them more mobile, easily ingested, easily inhaled, easily translocated,
and easily absorbed. They seem to cross all tissues, organs, including the brain(ACS, 2019).
Nanoplastics are either formed intentionally by companies as primary Nanoplastics (ACS, 2019;
Anumol, 2019) or formed by forces acting upon the large plastics as secondary microplastics.
Scientists opined that the large effects of plastics are mostly felt when they are in nano or
microscale and most of the public are unaware of the effects of Nanoplastics on biological
systems properties/ processes of Nanoplastics, and possible mitigations/ solutions (ACS, 2019).
This paper is discussing on the consequences of nanoplastics on biological systems in a laconic
fashion.
2. HARMFUL PROCESSES OF NANOPLASTICS
There are quite a number of processes or properties of plastics or NPs which aid in making
them readily harmful to biological systems. Some of these processes or properties are outlined
below:
Fragmentation - Due to UV light or heat, floating, wave, wind, animal bite, and other physical
forces, plastics tend to fragment into miniscule or smaller pieces to yield microplastics or NPs.
This property is responsible for making disvirgin or secondary nanoplastics, which can readily be
ingested, inhaled or absorbed by organisms and easily translocated within the body (GESAMP,
2015). Parable, polypropylene, and polythene undergo fragmentation (Rachid et al., 2018).
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Degradation - Breaking down of plastics into smaller pieces as a result of chemical action
(Sarkingobir et al., 2020).
Biodegradation - Exposure to UV rays, oxygen, high temperature, and microbes lead to

degradation of nanoplastics to yield water, carbondioxide, energy, and new biomass (UNEP
Frontiers, 2016; FAO, 2019).
Retention in biota -Nanoplastics can be taken up and retained in the biological systems, and
potentially transport them to other places or incorporate them in the food chain (UNEP Frontiers,
2016; Dris et al., 2018).
Ingestion - Several studies revealed that various biological systems take in nanoplastics,
example in fishes, mammals, birds, invertebrates (Leslie, 2014; Avio et al., 2016, ACS, 2018).
Uptake by tissues, cells - Some studies examined the occurrence of nanoplastics in tissues or
body-fluids. It was demonstrated that, plastics accumulate in the lysosomes, leading to
breakdown and expulsion of enzymes and death of cells or organisms (FAO, 2019; ACS,2020).
Vector to chemicals - Nanoplastics have the potential to shuttle chemicals along. They can
attached to hydrophobic or hydrophilic compounds and serve as conveying vehicle to chemicals
including persistent organic pollutants (POPs), polycyclic aromatic hydrocarbons(PAHs), and
other hydrocarbons (ACS, 2018; Revel et al., 2018). Other chemicals include: DDTs,
nonylphenol (NP), HCH, (PCBs), Polychlorinated biphenyls, polychlorinated dibenzofurans
(PCDF), polychlorinated dibenzo-p-dioxins(PCDD). They also covey microbes around (Wagner
et al., 2014; Avio et al., 2016, ACS, 2018;).
Inhalation - Humans inhale nanoplastics around them, which researchers believed, can be
ejected through coughing, or persist, leading to inflammation and can be potentially leads to
cancer (American Chemical society, 2018).
Leaching – Nanoplastics take in chemicals through sorption and these chemicals are easily leach
to the environments or organisms. They also leach their additives to the environment or
organisms. They also leach their additives to the surrounding. This leaching property is hasten by
UV light or heat (Gebeshuber, 2007; Prust et al., 2020). Major or most of plastics release toxic
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monomers linked to cancer and reproduction effects. It might be due to incomplete

polymerization, or heat/ UV/ mechanical actions. Many chemicals are found in plastics.
Bisphenol A is monomer in some plastics, which disrupts hormones, causes cardiovascular
disease, type 2 diabetes. Phthalates are industrial plasticizers found in plastics such as PVC.
They are extremely found in plastics such as personal care materials. Certain phthalates are
reported as endocrine disruptors (Science for Environment Policy, 2011).
Egestion - Some nanoplastics are released from the body in faeces or urine. From where it might
be taken by other organisms or persist in the environment. Example ammonium palmitoyl glycol
chitosan is taken orally, to the blood lymph, liver and bladder, then excreted via urine and faeces
(Galloway, n.d.). Metals strongly adsorb to plastics. Parable, Al, Fe, Cu, Pb, Zn, Cd, Cr,Ni,
etc(Avio et al., 2016).
Biomagnification - They tends to increase in concentration in the tissues of organisms at

successively higher levels in food chain (UNEP Frontiers, 2016).
Toxic monomers- There are many plastic monomers that are toxic to biological systems.Parable,
styrene increases cancer risk. It has reproductive effects, effects on CNS (depression, fatigue,
weakness, hearing loss etc), and effects on kidney, blood, stomach, and respiratory system
(Agency for Toxic Substances and diseases, 1992).
Vinyl chloride can cause effects on Nervous system, liver damage, and cancer (Agency for Toxic
Substances and diseases, 1997). PVC causes pollution due to chlorine, cancer causing vinyl
chloride, and ethylene dichloride ,dioxin, cadmium, lead, phthalates, tin etc (Centre for health
,Environment and justice / environmental Health Strategy Centre, 2004).
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Figure 1: Some features of nanoplastics
3. GENERAL HARMFUL EFFECTS OF NANOPLASTICS

A wide array of harmful effects of Nanoplastics on diversity of biological systems has been
observed by several studies. Some of these effects are: neurotoxicity, alteration of gene
expression, inflammation of gut, gills, liver, and kidney. Inflammation spurs oxidative stress and
aftermath consequences. Other effects are bioaccumulation in gills, intestine, liver, gall bladder,
gonads. It disturbed metabolism, destroy membranes, reduces fitness, and increased mortality
(Kogel et al., 2020; Prust et al., 2020). Particularly in humans, Nanoplastics lead to macrophage
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toxicity, decreased liver and lipid weights, increase mucus secretion(European Union, 2019;
Kogel et al., 2020).It is associated with asthma attack, heart disease, stroke, and respiratory
disease (NRDC, 2007). Their large surface area makes them more chemically reactive, and more
interactive with the body. They mimic biological components. They interact with DNA, leading
to inflammation, oxidative stress, and impairment of cell function (NDRC, 2007). Other effects
are mitochondrial depolarization, inhibition of activity efflux pump, inhibition of binding
cassette (ABC) transporter (Yong et al., 2020).
4. POSSIBLE REMEDIES/ SOLUTIONS

There are several solutions to control the consequences of nanoplastics. Some of the solutions
are identified below:
a. Ban unsafe, untested use of Nanoplastics. Unsafe, untested nanoplastics should be
avoided from human or environment
b. Conduct a full-lifecycle environment, health, and safety assessment before
commercialization(Natural Resources Defense Council, 2007)
c. Restriction of additives. Parable, bisphenol A is about to be banned in Europe, it has been
banned from baby bottles. Pthalates has been restricted in EU from certain plastic
products
d. Knowledge gap should be filled through extensive research, and development of
appropriate technologies for risk assessment (Science for Environment, 2011; Anumol,
2019)
e. Biotechnology use. It involves using microbes or microbial enzymes to treat plastic
waste. Typical microbes are Pseudomonas, Flavobacteria, Arthrobacter, Agromyces
(Ogunola, 2017)
f. Cleaning up- Plastic waste environment should be cleaned
g. Behavioural change- There is need for extensive awareness creation, so that leaders
become truthfully committed to address plastic threats. The public should fully be
educated to know the pros and cons of plastics, and act positively, also forced
governments to act well (Ogunola, 2017)
h. Taxation- Increased taxes for Nanoplastics is a step to reduce plastic use. It will lead to
hike in price, and low production and low patronage (Sarkingobir et al., 2020)
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5. CONCLUSION
Nanoplastics are advancements, but have negative consequences on biological systems.
REFERENCES
1. Agency for Toxic Substances and disease Registry (1992). Toxicological profile for
styrene.U.S. Public Health Service, US. department of Health and Human Services
Atlanta.
2. Agency for Toxic Substances and disease Registry (1997). Toxicological profile for vinyl
chloride. U.S. Public Health Service, US. department of Health and Human Services
Atlanta.
3. American Chemical Society (2018). Plastic continuation of the environment: sources,
fate, effects, and solutions.
4. Anumol T(2019). Micro and nanoplastics: A deep dive into a global issue.
5. Centre for Health, Environment and Justice / Environmental Health Strategy Centre
(2004). PVC bad news comes in: the poison plastic, health hazards and the looming waste
crisis.
6. European Union (2019). Nanoplastics damage marine creatures natural defences,
increasing lethal effects of POPs
7. Food and Agriculture Organization of the United Nations (2019). Microplastics in fishes
and aquaculture. What do we know? Should we be worried?
8. Galloway , T.S. (n.d.). Micro and nanoplastics and human health.
9. Gebeshuber, I.C. (2007). “ Social, health and ethical implications of nanotechnology.
Proc. 2nd Vienna International Conference Micro- and-nanotechnol. Vienna007.
10. GESAMP, Joint Group of Experts on the Scientific Aspects of Marine Environmental
Protection(2015). Sources, fate, and effects of microplastics in the marine environments:
A global assessment.
11. Ibrahim, M., Barau, L., Alhassan, M., Gidadawa, Z.S., Dan Galadima, H.,(2019).
Assessment of Environmental Impact of Solid Waste Generation and Disposal in Sokoto
Metropolis. International Journal of Scientific and Research Publications, 9(5):376-383.
12. Kogel, T., Bjorⱷy, Q., Toto, B., Bienfait, A.M., Sanden, M.(2020). Micro and nanoplastic
toxicity on aquatic life: Determining factors. Science of the Total Environment,
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709(2020):1-16.
13. Leslie, H.A. (2014). Macroplastics, microplastics and environmental impact. IVm Intitute
for Environmental Studies, University of Amsterdam.
14. Natural Resources Defense Council (2007). Nanotechnology’s invisible threat. Small
science, gi consequences. NRDC Issue paper may,2007.
15. Ogunola, O.S. (2017). Current advances in strategies to mitigate the impacts of
micro/nanoplastics: A review. Journal of Environmental and Analytical Toxicology,7(3):
1-4.
16. Prust, M., Meijer, J., Westerink, R.H.S. (2020). The plastic brain: neurotoxicity of micro
and nanoplastics. Particle and Fibre Toxicology, 17(24):1-16.
17. Rachid, D., Johnny, G., Vincent, R., Mohammed, S., Nicolas, R. et al (2018).
Microplastic contamination in an urban area: A case study in Greater Paris.
Environmental Chemistry, CSRO Publishing, 2015, pp.201510.10711EN14167.hal-
01134553
18. Revel, M., Chatel, A., Mouneyrac, C. (2018). Micro(nano) plastics: A threat to human
health? Current Opinion in Environmental Science and Health,1:17-23
19. Rist, and Hartmann, N.B. (2017). Aquatic ecotoxicity of microplastics and nanoparticles:
Lessons learned from engineered nanomaterials. In M. wagner and S. lambert (Eds).
Freshwater microplastics-Emerging Enviropnmental contaminants? Springer. The
handbook of Environmental Chemistry, vol 58.
20. Sarkingobir Y., Dikko M., Tambari U., Adamu AA., Salau IA., Gada MA(2020). The
dangers of plastic to public health: A review. NIPES Journal of Science and Technology
Research,2(2):195-200
21. Sarkingobir, Y., Muhammad, M.B., Gada, M.A., Sarkingobir, S. (2019). Land plastic
pollution in Sokoto: A case to worry about and threat to Sustainable Development Goals
(SDGs). A paper presented at a conference of National Tropical Biology Association
Federal University Dutse, Jigawa
22. Science for Environment Policy (2011). Plastic waste: Ecological and human health
impacts
23. Stapleton, P.A.(2019). Toxicological considerations of nano-sized plastics.
Environmental science, 6(5):367-378.
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24. UNEP Fronties (2016). Microplastics: Trouble in the food chain

25. Wagner, M., Scherer, C., Alvarez-Munoz, D., Brennholyt, N., Bourrain, X., Buchinger,
S., Fries, E., Grosbois C., Klasmeier J., Marti T., Rodriguez-Mozaz S., Urbatzka R.,
Vethaaak AD., Winther-Nielsen M., Reifferscheid, G.(2014). Microplastics in freshwater
ecosystems: What we know and what we need to know. Environmental Sciences Europe,
26(12):1-9.
26. Yong, C.Q.Y., Valiyaveetil, S., Tang, B.L. (2020). Toxicity of microplastics and
nanoplastics in mammalian systems. International Journal of Environmental Research
and Public Health, 17:1-24.
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CORONAVIRUS PANDEMIC AND RELIEF PROCEDURES: SUGGESTIONS FOR

MATERNAL AND KID WELLBEING AND NOURISHMENT
Shubham
Dyal Singh College, Karnal (132001) , Haryana

Mobile no.-8708075152
E-mail:- bhattishubham25@gmail.com
ABSTRACT
Covid illness 2019 (Coronavirus) keeps on attacking wellbeing and financial measurements
universally, remembering progress for maternal and youngster sustenance. In spite of the fact
that there has been center around increasing paces of youth squandering for the time being,
maternal and kid undernutrition rates are additionally prone to increment as a result of
Coronavirus and its effects on neediness, inclusion of fundamental intercessions, and admittance
to proper nutritious nourishments. Key areas at specific danger of breakdown or diminished
effectiveness in the wake of Coronavirus incorporate food frameworks, earnings, and social
security, medical care administrations for ladies and kids, and administrations and admittance to
clean water and sterilization. This survey features key zones of worry for maternal and youngster
nourishment during and in the fallout of Coronavirus while giving vital direction to nations in
their endeavors to decrease maternal and kid undernutrition. Established in learnings from the
models in Worldwide Wellbeing's Hindering Decrease Models venture, we give a bunch of
suggestions that range interests in areas that have supported immediate and roundabout effect on
sustenance. These incorporate mediations to reinforce the food-flexibly chain and decreasing
food uncertainty to help those at impending danger of food deficiencies. Different methodologies
could incorporate focused on social security net projects, installment deferrals, or tax cuts just as
reasonable money uphold programs for the most defenseless. Focusing on the most minimized
families in country populaces and metropolitan ghettos could be accomplished through sending
network wellbeing laborers and supporting ladies and network individuals. Network drove
disinfection projects could be critical to guaranteeing sound family conditions and decreasing
undernutrition. Furthermore, a few Coronavirus reaction estimates, for example, contact
following and self-detachment could likewise be misused for nourishment insurance. Worldwide
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wellbeing and enhancements in undernutrition will require governments, benefactors, and

improvement accomplices to restrategize and reprioritize ventures for the Coronavirus time, and
will require information driven dynamic, political will and responsibility, and global solidarity.
KEYWORDS: COVID-19 Pandemic, fundamental, sterilization, nourishment, administrations,

financial etc.
INTRODUCTION
As an exceptionally transmittable infection, Covid illness 2019 (Coronavirus) keeps on
assaulting the condition of the world's wellbeing and economy (1). Its effect additionally
underscores the restricted advancement we have made against noncommunicable infections
(NCDs). Youngsters and grown-ups with fundamental comorbidities, especially NCDs, for
example, diabetes, hypertension, undernutrition, and overweight/corpulence, are strikingly
helpless against genuine ailment and demise from Coronavirus (2). However, Coronavirus
reaction estimates, for example, self-detachment, social separating, and lockdowns of networks
can prompt helpless administration of key danger factors, for example, unfortunate eating
regimens and actual movement (2), and restricted admittance to preventive consideration in
essential consideration settings. Moreover, shaky financial conditions, limited travel and
admittance to medical care administrations, postponed inoculation timetables, and covering of
instructive offices further compound chronic frailty conditions for small kids, particularly in low-
and center pay nations (LMICs) (3). There is huge worry that Coronavirus reactions have
negatively affected the healthful status of ladies and youngsters, and that these could exacerbate
over the long run. An ongoing demonstrating activity of different assessments of the likely effect
of Coronavirus related financial decay, food instability, and interference of projects of network
based recognition and the board of ailing health proposes that the predominance of squandering
could increment by 10–half with an overabundance of ∼40,000–2,000,000 kid passings (4).
We accept that these extended nourishment impacts of the worldwide pandemic could
well be belittles, as they neglect to consider the likely impact on maternal sustenance,
micronutrient inadequacies, and intrauterine development just as downstream effects on maternal
and kid wellbeing programs that can affect straight development and youth hindering. This is
awful since the world has made a few, but moderate, progress in diminishing youth hindering
throughout the most recent decade. Current evaluations show that 149 million kids under 5 y are
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hindered; a decrease from 166 million of every 2012 yet at the same time a long way from the
necessary worldwide focuses for progress (5). Coronavirus presently takes steps to stop or
converse increases significantly further. On the off chance that unaddressed, the consequences
for straight development in youngsters and resulting hindering could be significantly more
important than momentary impacts of undernutrition.
DANGER COMPONENTS FOR UNDERNUTRITION WITH REGARDS TO

CORONAVIRUS
Areas basic to diminishing youth undernutrition at specific danger of breakdown or decreased

productivity because of far reaching effect of Coronavirus are summed up beneath and in .
• Food weakness and low quality eating regimens
• Building strong food frameworks during Coronavirus requires inventive setting explicit
interest and gracefully side activities. Food flexibly chains (FSCs) are quite compelling, since
80% of all nourishments burned-through in Africa and Asia are currently subject to these
business sectors (6). In spite of being ostensibly "excluded" from lockdowns, Coronavirus can
have immediate and circuitous effects on FSC work in LMICs, particularly the casual areas.
While direct effects, through terminations of eateries and limitations on merchants, speak to a
little portion of the absolute food economy in metropolitan settings, the effect on country
markets could be a lot more noteworthy Furthermore, circuitous effects because of joblessness
and falling livelihoods of day by day wage workers and industry laborers have negatively
affected individuals in LMIC settings (7). Further exacerbating this is the issue of food valuing.
Limitations on systems for creation and conveyance may drive up cost, while dread of
deficiencies could drive theoretical storing (8). Loss of family unit pay opens weak families to
value spikes and food deficiencies, while low farming efficiency and breaks in the food import–
trade framework upset nearby food markets and private ventures (9).
• Additionally, given restricted admittance to new deliver, youngsters and families might be
bound to depend on less expensive and more available prepared and prepackaged, high-sodium,
and less-nutritious nourishments (10), with injurious wellbeing results.
• Reduced pay and restricted budgetary assets
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• COVID-19 has driven great many family units into monetary despondency and has been
portrayed as more deadly than the 2008 worldwide money related emergency (11). Oxfam
predicts that a large portion of a billion people could be driven into neediness (12), while the
World Bank battles that an extra 40–60 million individuals could be driven into extraordinary
destitution (13). The interference of existing social wellbeing nets, particularly for ladies, is a test
in numerous LMICs battling with Coronavirus as assets are redirected to prompt needs
compounded by restricted versatility and admittance to administrations.
• Limited care and limited wellbeing administrations
• Given overburdened wellbeing frameworks, limited travel, and changing needs at the essential
consideration level, admittance to routine wellbeing administrations for ladies and youngsters
has endured massively. While nature of care was a continuous test preceding Coronavirus (14),
in its present status and forward for quite a long time focused on endeavors for top notch medical
services for those in the most need will probably assume a lower priority. Thusly, the wellbeing
and danger of undernutrition in moms and their youngsters may increment drastically,
particularly if current conditions endure long haul. In Pakistan, accessible information from
locale wellbeing frameworks show an emotional drop in access for and arrangement of antenatal
consideration administrations (ZA Bhutta, individual correspondence, 2020), and others have
featured the significance of the neglected requirement for psychological well-being
administrations and intercessions (15). As has been demonstrated by Joined Countries Populace
Asset (UNFPA) (15), decreased admittance to family-arranging administrations and authorized
imprisonment of families is extended to prompt 7 million unintended births in probably the least
fortunate nations of the world. Persevering interruptions to normal and imperative maternal
consideration and sustenance could prompt antagonistic fetal results including preterm birth, low
birth weight, and little for-gestational-age babies.
• Interrupted instruction for youngsters and grown-ups
• Educational offices, including essential, optional, postsecondary, and particular preparing

organizations, have been covered totally worldwide in the wake of Coronavirus (16, 17). One of
the significant impacts of Coronavirus has been on fueling imbalances in instruction. Much has
been made of elective types of learning, for example, online homerooms, electronic courses, and
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self-teaching, however these are distant to most kids in LMICs. Ladies and young ladies, who
regularly experience the most noteworthy paces of ignorance and school nonconformists in
LMICs, are yet additionally crippled and distraught. The advantages of general and concentrated
wellbeing and sustenance schooling to improve maternal nourishment and lessening
intergenerational youth hindering are unquestionable, having been indicated reliably in hindering
contextual analyses (18–20). An extra mishap has been the interference of school sustenance
programs, the pillar of tending to food weakness in the absolute most unfortunate areas of the
populace.
• Unhealthy family climate
• Given redirected assets and needs, building sheltered and sound family and network
conditions, especially as identified with clean water, proper disinfection, and cleanliness
(WASH), may fall behind on nation plans. But at this point like never before, WASH mediations
are fundamental to ensuring human wellbeing and forestalling undernutrition . For example, in
metropolitan ghettos (the absolute most weak networks), lockdowns and restricted versatility
have affected admittance to clean water and safe disinfection administrations. Given the idea of
Coronavirus transmission, this could bring about deadly episodes of irresistible illnesses.
Coronavirus direct impacts on essential, fundamental, and prompt drivers of intense and
ongoing ailing health. Coronavirus, Covid infection 2019.
We accept that nations can address these unprecedented nourishment chances over the
continuum of moms, infants, youngsters, and youths by tending to determinants and actualizing
proof educated techniques for activity.
This account is pointed toward auditing key territories of worry for supporting maternal and
youngster nourishment progress during and in the result of Coronavirus, while giving key
direction to nations to keep making progress in lessening maternal and kid undernutrition while
engaging Coronavirus. As our investigation into hindering decrease models has illustrated,
hindering advancement in LMICs has been driven by a multifactorial arrangement of interests in
areas that have immediate and roundabout effects on sustenance , the majority of which are
amazingly applicable in the current Coronavirus emergency and must be proceeded.
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Ventures to organize both inside and past the wellbeing area to relieve Coronavirus outcomes
on nourishment. Coronavirus, Covid sickness 2019; MNCH, maternal and kid wellbeing.
PATH FORWARD
Our models underscore various instances of high-sway methodologies both inside and outside
a nation's conventional wellbeing framework. These models were information driven and
empowered by solid, centered nation authority, productive financing, and successful
organizations . We accept that a similar methodology is required inside LMICs to address the
wholesome results of Coronavirus relief procedures.
The condition of the world and our aggregate reaction to Coronavirus is consistently
advancing as new data is gotten. By the by, starting perceptions across various nations and
settings, alongside key exercises from nations overseeing through different emergencies
previously, recommend that we organize the accompanying ways to deal with address and
forestall worsening maternal and youngster undernutrition:
• Food uncertainty intercessions
• Given the variety of food climate and security challenges experienced by LMICs during
Coronavirus, arrangements must be setting explicit. Exercises from many hindering decrease
model nations could be valuable. In the Kyrgyz Republic, for example, the remarkable monetary
breakdown after the disintegration of the Soviet Association made new open doors for preparing
the horticultural area to drive financial recuperation. A scope of revolutionary agrarian changes
zeroed in on renewing establishments for land, animals, capital, and work, while simultaneously,
moving area possession from the state to private families was considered among the most crucial
driving components of hindering decrease in Kyrgyz Republic somewhere in the range of 1990
and 2014 . While agrarian land changes zeroed in on moving area possession and embracing
inventive/effective farming practices may yield profits on undernutrition in the long haul, quick
arrangements additionally have esteem. One of Ethiopia's answers for food uncertainty (i.e., the
Profitable Wellbeing Net Program) was pointed toward giving crisis food help to 15 million
people powerless against food instability and was viewed as critical to the nation's hindering
decrease story (18). Such long-and momentary arrangements tending to both gracefully and
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request side difficulties could be considered for sustenance security in Coronavirus influenced
nations.
• Social security programs
• The prioritization of endeavors to give monetary security by governments to their in danger

populaces (e.g., through creative and focused on social wellbeing net projects, installment
deferrals, or tax reductions) is fundamental to forestalling budgetary breakdown of weak
families. Social-insurance programs are progressively becoming the dominant focal point in
strategy exchanges for handling neediness, weakness, and social avoidance. A few model
nations, prominently Peru (20) and the Kyrgyz Republic , utilized effective money related
motivator based models as a methods for giving social wellbeing nets to arriving at
underestimated and weak populaces. In Peru, for example, the Juntos restrictive money move
program furnished families with a fixed month to month money move (∼$30 USD) to consent to
fundamental instruction, wellbeing, and sustenance administrations for kids. This was matched
with solid information the board frameworks that took into consideration recognizable proof of
weak populaces and viable focusing to guarantee that assets were dispensed successfully. The
Kyrgyz Republic's Month to month Advantage for Helpless Families with Youngsters Program
is a similar to basic social-insurance plot that was discovered to be remarkably critical to
hindering decrease in the nation. In the present Coronavirus climate, such frameworks in Peru,
Kyrgyz Republic, and numerous different nations can be utilized to expand on and improve
social and financial assurance for weak families, and thusly forestall medical affliction and
persistent undernutrition in youngsters.
• Access to medical care
• As has been appeared in a few hindering decrease model nations, admittance to medical care
for even the most far off and difficult to-arrive at populaces can occur with a powerful network
wellbeing augmentation framework. Ethiopia's wellbeing expansion laborers (Cuts) (18) and
Nepal's female network wellbeing volunteers (FCHVs) (19) exhibit effective models of
activating network wellbeing laborers (CHWs; who get fundamental preparing and wares) to
convey antibodies, dietary enhancements, wellbeing and nourishment schooling, and even
regenerative, maternal, and infant care. The current proposals are to compensate such CHWs
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instead of depend on unadulterated volunteerism. In the midst of the Coronavirus emergencies,

while the essential medical care framework may not be completely useful and supplies short,
governments could consider approaching existing CHW units to reprioritize their assignments
and oblige rising maternal, kid wellbeing, and sustenance screening in networks. These CHWs
are likewise key to restoring programs for network based administration of hunger. Governments
could likewise put resources into sending extra wellbeing laborers and boosting current
specialists to keep conveying great basic mediations to families (e.g., antibodies, antenatal
consideration, references) and give basic correspondence identified with Coronavirus readiness
and emergency. Where people group wellbeing expansion programs at present don't exist,
nations might need to think about steering or receiving such a program to enhance essential
medical care, as a short-or long haul arrangement.
• Educational programs
• In the wake of shut conventional instruction frameworks, nations could activate casual
establishments, for example, CHWs and ladies' and network uphold gatherings to convey
wellbeing and general training. These frameworks are as of now set up in numerous LMICs and
could be renewed and repurposed for proceeding with instruction. A few hindering decrease
model nations have indicated the expected utility and effect of these systems on hindering
decrease. Having gained from their involvement in Ebola, Senegal's CHW program has
demonstrated to be a successful system for imparting wellbeing best practices to the network.
Nepal's FCHV (19) and Ethiopia's Slash (18) programs have likewise had profoundly effective
wellbeing and nourishment guiding parts. The Kyrgyz Republic utilized ladies' care groups in
networks as a way to keep refreshed on the advancing wellbeing circumstance and offer
information , a model that could keep on being developed.
• Safe and sound family/network conditions
• Ensuring safe water access and suitable sterilization and cleanliness rehearses is basic to the
Coronavirus control plan and wellbeing results past. While giving infrastructural backing to
families and networks (e.g., through building wells, network pipes, lavatories) is basic, it might
tumble off momentary plans as handwashing and cleanliness crusades outweigh everything else.
Exercises from model nations propose that high-sway, ease network assembly endeavors could
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assume a vital function in establishing a solid climate by diminishing open poop and
empowering sterile practices and have been connected to hindering decrease. The People group
Drove All out Sterilization (CLTS) programs in Nepal (19), Ethiopia (18), and Senegal center
around social change to make open-poo free towns. The projects trigger the network's longing
for aggregate change through empowering advancement and setting explicit arrangements while
cultivating a feeling of network proprietorship. The CLTS programs in model nations, for
example, Nepal have notably affected youth hindering decrease.
Past intercessions focusing on explicit difficulties for youth hindering, numerous continuous
Coronavirus reaction measures could twofold as occasions to address other wellbeing and
prosperity needs, for example, unhealthiness anticipation and the board in LMICs. Governments,
contributors, and improvement accomplices during Coronavirus reaction strategy and subsidizing
exchanges ought to plan on cost and framework efficiencies for focusing on more extensive
wellbeing and sustenance objectives inside their Coronavirus reaction plans.
CONCLUSION
The Coronavirus pandemic has tossed the world into a phenomenal emergency, battling a
microbe that could well be with us for quite a while to come. As nations reel from the stun of
huge scope lockdowns to a steady re-visitation of routineness, the progress will be moderate and
the new ordinary altogether different from an earlier time. Shielding the wellbeing and
nourishment of weak ladies and kids is a key strategy reaction and must be founded on the best
proof of what works, so that gains in endurance and ladies' and youngsters' wellbeing and
sustenance are not turned around. Governments, givers, and improvement accomplices will
together need to restrategize and reprioritize speculations for the Coronavirus period utilizing
information driven dynamic. Successful execution of procedures will require cash, political will,
and duty, and worldwide solidarity; these will be crucial drivers, making sure about Coronavirus
explicit gains as well as by and large assurance of worldwide wellbeing and enhancements in
undernutrition.
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statements/dont-let-youngsters be-covered up casualties Coronavirus pandemic.
4. Roberton T, Carter ED, Chou VB, Stegmuller AR, Jackson BD, Hat Y, Sawadogo-Lewis
T, Walker N. Early gauges of the backhanded impacts of the Coronavirus pandemic on
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https://doi.org/10.1016/S2214-109X(20)302291.
5. Global Sustenance Report. Worldwide Sustenance Report 2020. [cited May 20, 2020]
[Internet]. Accessible from: https://globalnutritionreport.org/reports/2020-worldwide
nourishment report/.
6. Reardon T, Echeverria R, Berdegué J, Minten B, Liverpool-Tasie S, Tschirley D,
Zilberman D. Fast change of food frameworks in creating locales: featuring the function
of farming exploration and advancements. Agric Syst. 2019;172:47–59.
7. Thurlow J. Coronavirus lockdowns are forcing generous financial expenses on nations in
Africa. [Internet]. Worldwide Food Strategy Exploration Organization; 2020; [cited May
20, 2020]. Accessible from: https://www.ifpri.org/blog/Coronavirus lockdowns-are-
forcing considerable monetary costs-nations africa.
8. Reardon T, Bellemare MF, Zilberman D. How Coronavirus may upset food gracefully
chains in non-industrial nations. [Internet]. Worldwide Food Strategy Exploration
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https://www.ifpri.org/blog/how-Coronavirus may-disturb food-gracefully chains-

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[cited May 12, 2020]. Accessible from:
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Endris BS, Islam M, Keats E et al. Drivers of hindering decrease in Ethiopia: a nation
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Rappaport A, Tasic H, Vaivada T et al. Drivers of hindering decrease in Peru: a nation
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APPLICATIONS OF BIOTECHNOLOGY – A REVIEW

PhD Scholar, Pachaiyappa’s College, Chennai-30
meerarasul7@gmail.com
ABSTRACT:
This paper targeted at evaluating biotechnology with relevant to its application. Major areas
of applications identified in the literature are environment, medicine, agriculture, food processing
and industry. It was observed that the areas and scope of application of biotechnology would
broaden with respect to advancement in science. It was concluded that as the scope of
biotechnology application elaborating ing. Research works should focus on the risks and
challenges identified, especially in agricultural (Genetically modified foods) and medical
applications.
KEYWORDS: Biotechnology application, environment, medicine, agriculture, food processing
INTRODUCTION
Biotechnology is technology that utilizes biological systems, living organisms or parts of this
to develop or create different products. Brewing and baking bread are examples of processes that
fall within the Concept of Biotechnology. According to Bull. Holland Lilly (1982),
biotechnology is defined as the application of scientific and engineering principles to the
processing of materials by biological agents to provide goods and services. In similar definition
by Boyer (2016), it is the technique of using live organisms or enzymes from organisms to
produce products and processes useful to human. From the forgoing, one could think of making
of wine using microbe based processes as a form of biotechnology. However, biotechnology is
used in a restricted sense to refer to the use of genetically modified organisms to produce
products and processes for mankind. The genetic modification is achieved through genetic
engineering, which is a process to alter the chemistry of genetic material (DNA or RNA). Other
technique in biotechnology is the maintenance of sterile ambience chemical engineering
processes to enable growth of only the desired microbe in large quantities for the manufacture of
biotechnological products like antibiotics, vaccines, enzymes, etc. (Boyer. 2016). Many
processes are involved in biotechnology which include but not limited to In Vitro Fertilization
(IVF) applicable in medicine and the use of micro-organisms in the prevention. treatment and
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monitoring environmental pollution. There are many areas biotechnology is applicable: notable
ones are health care (medical), crop production and agriculture, non-food (industrial) uses of
crops and other products (e.g. biodegradable plastics, vegetable oil, biofuels), and environmental
uses.
VARIOUS APPLICATIONS OF BIOTECHNOLOGY
APPLICATIONS OF BIOTECHNOLOGY IN ENVIRONMENT

Application of biotechnology in the environment is the process of applying microorganisms
to improve environmental quality. This process is often termed as environmental biotechnology.
The areas of application are conversion of organic wastes, environmental bioremediation of
hazardous contaminants, environmental protection and monitoring (Maria and Violeta 2013 and
Gavrilescu, 2010). Conversion of organic wastes into useful bio resources is possible through
biotechnological processes using microorganisms. The organic waste sources under
consideration are plants, agricultural wastes and municipal residues. These wastes of plant origin
compose of lignin, cellulose and hemicellulose. For instance, conversion of organic waste into
nutritive biomass involves the conversion of cellulose to high calorific food or feeds using
cellulolytic bacterial. Also, conversion of wastes into bio energy such as bio fuel involves the use
of microorganisms such as fungi.
Environmental bioremediation is the process of reducing or eliminating pollutants in the
environment through the application of biotechnology in the form of biotreatment. According to
Gavrilescu 2010, biotreatment /bioremediation methods are applied to remove, degrade or
detoxify pollution in environmental media such as water, soil, air, and wastes. Microbes such as
yeasts, fungi, protozoa, unicellular plants, rotifers and bacteria are used in bioremediation
process because most of them have the potential to degrade most hazardous and recalcitrant
chemical pollutants in the environment.
Environmental protection is possible through environmental biotechnology. For example,
carbon dioxide mitigation such as CO, recovery from the pollutant gas discharges from industries
or on-site CO, fixation by bioprocess using cyanobacteria has been reported in the literature
(Hitoshi et al, 2010). The removal or mitigation of CO, from the atmosphere or gaseous pollutant
is necessary to prevent global warming and this practice is achievable by the application of
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biotechnology. Furthermore, the removed CO, can be converted into biodegradable plastics as
substitutes for petroleum-based plastic
Environmental monitoring is possible with the use of biosensors in biotechnology.

Biosensors can be used to measure pollutant levels and can detect pollutants such as heavy
metals. herbicides, pesticides and organic compounds as detailed in Gavrilescu (2010).
APPLICATIONS OF BIOTECHNOLOGY IN MEDICINE

The goal of biotechnology in medicine is to fight and cure diseases. Consequently
biotechnology is applicable in the following areas of medicine: production of drugs and
therapeutics, genetically modified organism, analysis of genes in genetic diseases, corrections of
genetic defections, etc.Other areas of application in medicine can be found in Biotechon web
(2018) and Mohammad and Narasu (2013. rDNA technology was applied in therapeutic
application by generating genetically engineered insulin for man. In 1983, Eli Lilly, an American
company prepared 2 DNA sequences coding for chains A & B. Human insulin consists of two
short Polypeptide chains A & B being linked by disulphide bridges.In man, Insulin secreted as
Prohormone containing C peptides that is removed during maturation. In rDNA technique,
insulin could be generated by preparing two separate DNA sequences of A & B chain which are
incorporated into plasmids of E. coli to produce insulin chains.
GENE THERAPY:
• Gene therapy involves correction of the gene defects in child or embryo.
• Adenosine deaminase deficiency is a kind of immuno-disorder caused by deletion of gene

coding for ADA.
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• It can be cured by bone marrow transplantation or enzyme replacement therapy.
• A functional ADA-cDNA(through Retrovirus) is introduced in lymphocyte culture for genetic

infusion and transferred to the patient body normal functioning.
APPLICATIONS OF BIOTECHNOLOGY IN MOLECULAR DIAGNOSIS
Early & accurate detection of diseases substituting conventional diagnostic tecniques may
be done by following methods: PCR (Polymerase chain reaction): Short stretches of pathogenic
genome is amplified for detection of suspected AIDS, Cancer or genetic disorder. ELISA
(Enzyme Linked Immunosorbent Assay) used to detect AIDS based on detection of antibodies
produced against antigen of pathogen
Transgenic Animals
Animals with manipulated genes or a foreign gene to be expressed are called as transgenic
animals. They are useful-
1. To know how genes contribute to development of disease.
2. To use proteins for treatment of disease.
3. To verify vaccine and chemical safety.
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Biopiracy :-
Some organizations and multinational companies exploit or patent bioresources of other

nations without proper authorization. Indian patent bill is there to prevent such unauthorized
exploitation.
GEAC- For validity of GM research and the safety of introducing GM organism.
Polymerase Chain Reaction(PCR) Enzyme Linked ImmunoSorbent Assay(ELISA)
APPLICATIONS OF BIOTECHNOLOGY IN WASTE MANAGEMENT
Methods based on biotechnology in wastewater treatment are activated sludge,trickling

filters, oxidation ponds, bio filters and anaerobic treatment. Furthermore, solid waste composting
techniques, biotrickling filters and biosorption are the examples of biotechnology applications
in waste management. Dec 16, 2015.
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APPLICATIONS OF BIOTECHNOLOGY IN ENERGY PRODUCTION

It is logical to imagine the future contribution of biotechnology to world energy
production may increase not only in the area of biofuel production, but also in
petroleum production, petroleum upgrading, biogas production, chemical production, crop
improvement, bioremediation, microbiologically influenced corrosion, ...Oct 20, 2020.
APPLICATIONS OF BIOTECHNOLOGY IN AGRICULTURE

Application of biotechnology in agriculture offers tremendous benefits which include
increased crop productivity, enhanced crop protection, improvement in food processing, caution
should be entertained. Improved nutritional value, better flavour, etc., as detailed in Wieczorek
(2003). Genetically modified organisms (GMO)-Plants, bacteria, fungi, animals whose genes are
altered by manipulation.
Transgenic crops(GMO) :-Crops contain or express one or more useful foreign genes.
Advantages :-
i) More tolerant to stresses (heat, cold, drought).
ii) Pest resistants GM crops reduce the use of Chemical pesticides. Eg- BT
Cotton
iii) Reduced post harvest losses. Eg- Flavr savr tomato.
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iv) Enhance nutritional value of food. eg.- Golden Rice (Vitamin A enriched)
v) Increased efficiency of mineral use.
APPLICATIONS OF BIOTECHNOLOGY IN FOOD PROCESSING:

Applications of biotechnology in food processing include fermentation bioprocess, the use of
food additives and processing aids used in food formulations Such products are enzymes, amino
acids, vitamins, organic acids, certain carbohydrates and flavouring agents produce using
genetically modified micro-organism as highlighted by FAO, 2010 and Jasia et.al.. 2010.
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APPLICATION OF BIOTECHNOLOGY IN INDUSTRY.
Industrial biotechnology is the application of biotechnology for industrial purposes. It refers

to the bio-process of crops and other products for non-food uses, that is, industrial uses.Bio
processes include industrial fermentation, the use of cells or micro-
organisms or enzymes to produce industrially useful products such as chemicals, feeds,

detergents, paper, bio-plastics and so on (Wikipedia. 2018).By producing products through bio-
processes, industrial biotechnology is a way of mitigating against petrochemical based economy
and encouraging sustainable economy.
CONCLUSION
Biotechnology was assessed with respect to its application. The assessment shows that
biotechnology is applicable in the environment, medicine, agriculture, food processing and
industry. It was observed that the areas and scope of application will continue to broaden as
science advances. It was concluded that biotechnology research works should be directed
towards the risks and challenges identified, especially in agricultural applications.
REFERENCES:
1. Adeogun B.K. (2018): Biotechnology and its Applications: A Review Biotecho Web (2018):
Application of Biotechnology. http://www.biotechonweb.com/Apnlications-of-Biotech html
Available online.
2. Boyer, H. (2016): Biotechnology: Principles and Processes. National Council of Educational
Research and Training. New Delhi Available on-line @ ncert.nic.in/ncerts/Viebol11.pdf.
ISBN: 978-81-948129-1-3 Page 110

3. Bull, A.T., Holt. G. and Lilly. M.D. (1982); Biotechnology: International Perspectives.
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on-line Jasonwww.intechapen.com.
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Agricultural Forestry Wastes into Nutritive Biomass. In Environmental Biotechnology -
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9. Mohammad, Z., and Narasu, M.I. (2013) A reviewer article: Biotechnology Applications in
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processing, wfeo On line at: http://www.wfeo-cee.org/news/v27n10pg2,.html.
11. Tehmeena, A., Fiza, N. and Rehana, S. (2017): Application of Biotechnology in Food
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4(12) December, 2017. Available on-line @www.ijetsr.com.
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13. Wieczorek A. (2013): Use of Biotechnology in Agriculture Benefits and
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(CTAHR), Hawaii. Available on-line @ www.ctahr hawaii edu.
14. Wikipedia (2018); Biotechnology. Wikimedia Foundation, Inc. Available on
lineat:https://en.m.wikipedia.org/wiki/Biotechnology.
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APPLICATION OF BIOINFORMATICS TO NANOTECHNOLOGY

Dr.K.Shoba
Assistant Professor, Department of Biochemistry, DKM College for women (Autonomous),
Vellore.
Email Id: danishoba@gmail.com
ABSTRACT
Bioinformatics consists of biology, mathematics and computer science and is an
interdisciplinary field of science. The use of information technology to handle biological data
that helps to decipher plant genomes is bioinformatics. In the emerging field of personalised
medicine, recent developments in bio-computing and nano-technology have accelerated the
development of novel biomarkers. Personalized medication from each individual's molecular
profile deals with disease diagnosis and therapy. Personalized medicine is also known as
predictive medicine, which uses genetic / molecular data to predict the growth, progression, and
clinical outcome of diseases. Recent advances in bio computing have served as the basis for the
detection of a nano particle with multiplexed probes. Together, we have compared bio-molecular
signatures with clinical findings and have also discussed an emerging area called bio-nano-
informatics to propose an individual treatment for cancer and other diseases.
KEYWORDS: Bioinformatics, Bio-computing, Nano-technology, Personalized medicine and
Nano particle.
INTRODUCTION
In 1970, Paulien Hogeweg and Ben Hesper coined the term Bioinformatics as a study of
machine processes in biotic systems. Computational management and analysis of biological
information (genes, genomes, proteins, cells, ecological systems, medical information, robotics,
artificial intelligence, etc.) is discussed in bioinformatics. Bioinformatics has been described by
the National Center for Biotechnology Information (NCBI 2001) as the area of science in which
biology, computer science, and information technology are used. At the Pasteur Institute, Fredj
Tekaia defines bioinformatics as mathematical, statistical and computational methods aimed at
solving biological problems by using DNA and amino acid sequences and related data. Hundreds
of microbial genomes have been sequenced and archived at Gen Bank for public study since the
sequencing of the first full microbial genome of Haemophilus influenzae in 1995. The vast
amount of knowledge produced by projects for genome sequencing is becoming unmanageable.
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Bioinformatics has emerged as a crucial science field that has been facilitating biological
discoveries for over a decade. Without the use of bioinformatics software, it is literally
impossible to collect, process, analyse and interpret the enormous quantities of data available,
especially after entire genome sequencing projects. For the farming world, sequencing the
genomes of plants and animals would have tremendous benefits.
To scan for the genes inside these genomes and to elucidate their roles, bioinformatics
techniques can be used. This particular genetic knowledge could then be used to grow stronger
crops that are resistant to drought, disease and insects, and to improve quality. In agriculture, it
helps to improve nutritional quality, rational improvement of plants, waste cleaning, climate
change studies, and the production of varieties of drought resistance (Dahiya and Lata, 2017) in
insect resistance. In addition to this, biotechnology, antibiotic resistance and forensic study of
bacteria, comparative studies, evolutionary studies and veterinary sciences also play an important
role.
Many computing tools and applications have appeared in the context of biomedicine over the
past five decades, leading to interdisciplinary fields such as medical informatics, bioinformatics
and others. These biomedical-related computer science fields cover a broad variety of theoretical
and technical approaches to solving complex problems, including, but not limited to, methods of
data and information integration, biomedical ontology and vocabulary, data and text mining,
interoperability of systems, interoperability of systems, DNA and RNA sequencing, support for
medical decision-making, prediction of relationships between gene mutations and diseases,
development of data representation and sharing standards, or the development of computer
methods and tools for multilevel data integration and multi-scale biomedical system simulation.
Data scientists have successfully contributed to these fields, leading to excellent outcomes such
as Human Genome and other omics initiatives, Clinical practise computerization or the
development of decision-support computerised systems. In recent decades, the authors have been
involved in computer science research, supporting a variety of ventures and pioneering important
examples, such as medical expert systems, among others.
A challenging new field, nanomedicine, that promises to deliver scientific and technological
breakthroughs that could transform medicine, is beginning to receive attention from the scientific
community, including computer scientists, after several decades of research on such biomedical
systems. Technical advances such as molecular cloning development, Sanger sequencing, PCR,
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microarrays of oligonucleotides and, more recently, the development of a range of so-called

next-generation sequencing (NGS) platforms have actually revolutionised translational research
and cancer research in particular. Now, scientists may obtain a genome-wide view of the
expression of cancer genes useful for finding new biomarkers of cancer for more precise
diagnosis and prognosis, and tracking treatment effectiveness. For example, microRNA
expression signatures have been shown to provide a more accurate way of classifying cancer
subtypes than transcriptome profiling, allowing different stages of tumour progression to be
identified, effectively opening up the field of personalised medicine (in which disease detection,
diagnosis, and therapy are tailored to the molecular profile of each individual) and predictive
Medicine (in which genetic and molecular data are used to predict the development of diseases,
progression and clinical outcome).
With the support of organisations such as the US National Science Foundation, the National
Cancer Institute and the European Commission, nanoinformatics has only recently emerged to
resolve these issues. In this modern sense, nanoinformatics (biomedical or nanomedical) refers to
the use of computer technology to analyse and process knowledge on the structure and
physicochemical characteristics of nanoparticles and nanomaterials, their interaction with
environments, and their application for nanomedicine. In a time when genomic and personalised
medicine are still gaining acceptance, such new applications arise and promise additional
potential insights for biomedicine. In biomedical practise and science, given the rapidly
advancing expansion of all informatics issues emerging between molecular biology and systems
biology (bioinformatics and computational biology) and public health (for public health
bioinformatics), we can ask if existing informatics applications, such as bioinformatics or
medical informatics, may also be ready to address this new area. At a distinct nano level, which
means substantial physical and chemical variations, the latter is regarded by many as the new
frontier in medicine, presenting major challenges for science, medical practise and economic
consequences due to novel toxic-therapeutic tradeoffs at the nano level. In this regard, we will
remember what happened between 1995 and 2001, when bioinformatics led to the early
completion of the Human Genome Project, and here it can be concluded that computer science
would also be necessary for the development of nanomedicine to advance. The possibilities of
reusing informatics methods, techniques, data and lessons learned in addition to new computing
methods and tools can well lead to speeding up the development of nanomedicine.
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Nanomedicine contains a wide range of significant practical and scientific impact subjects, such
as early detection of diseases such as cancer, the ability to achieve extremely precise targets
within the body, modern molecular imaging methods based on the optical properties of
nanoparticles, very low dose drug delivery control methods, diagnostic and therapeutic
nanorobots, and therapeutic nanorobots and innovative approaches to overcoming the limits of
solubility of new or existing medicines. A large number of reports can be downloaded by readers
for further reference.
SCIENTIFIC CONTENT AND PRIORITIES FOR INFORMATICS
Important attempts have been made during the past decade to correlate molecular and clinical
evidence with scientific discovery, leading to significant achievements. However, the semantic
heterogeneity, the inherent difficulty and confusion associated with the linkage of information
from these various biological levels have shown that it is difficult to reach the original standards
of applying information and experience from omics to clinical practise and prognosis. These
difficulties can be expected to increase with nanomedicine, but their potential is also excellent.
At the nanoscale, very distinctive quantum effects and size-related phenomena take place
from a scientific perspective. Physicists and chemists have shown that bridging orders of
magnitude in size poses additional theoretical difficulties in nanoscience. Although the
fundamental quantum physics laws necessary at the nanoscale are well established, additional
models and instruments are needed to quantify and compare the interactions between
nanoparticles and biomolecules with the results of these models with experimental
measurements.
For example, quantum dot research or the production of new drug delivery nanoparticles
benefit from new data, knowledge, theory and models of changes in physical characteristics of
nanoscale versus bulk materials that was not previously accessible or understood. Many
molecular tools in use for bioinformatics cannot be specifically applied in this context, while
others can, and as a consequence, major contributions from the field of multi-scale modelling
and simulation hold the promise of extending the reach of biomedical engineering and
informatics in fields such as imaging.
DATA AND INFORMATION COMBINING
For the advancement of science, structuring data in nanomedicine is important. In 2007 [in
MeSH, the controlled vocabulary used in Pubmed for organising and indexing biomedical
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literature], the word 'nanoparticle' was introduced. New taxonomies and ontologies have been
developed, such as the Nanomedicine Taxonomy (NT) and Nanoparticle Ontology (NPO). Other
projects include an ontology for the exploration of new nanomaterials, the Ontology Nanotech
Database, as well as the introduction of applications to map between various ontologies, such as
BiomedGT. Efforts such as the NPO may contribute to the collection of data, classification of
information, scanning and retrieval, data and text mining. Professionals may access information
from previous research by annotating nanoparticles and discover knowledge that may reside in
such data. Such work, including those included in the Open Biological and Biomedical
Ontologies (OBO) Foundry will extend the reach of biomedical ontology.
SKILLED PRACTISE SUPPORT RESOURCES
Criteria such as scale, shape, topology, composition, pharmacokinetics, biologic activity or
toxicity should be used to select nanomaterials for medical diagnosis and therapy. The required
nanoinformatics methods and tools will require extensions of those already developed, such as
omics and clinical data integration tools, for example. While nanoinformatics is in its initial
development phase, there will be great opportunities for computer specialists to contribute
nanomedicine expertise and applications.
CONCLUSION
To solve complex biological problems, bioinformatics puts mathematics, statistics, and
computer science and information technology together. In every area of life, the methods of
bioinformatics are useful. In agriculture, a variety of benefits are offered by genome sequencing
of plants and animals. The ultimate aim of bioinformatics is to incorporate large-scale data into
different developmental processes to understand the molecular mechanism involved. In the
development of Climate Smart agricultural crops, this understanding will benefit. When studying
this new area of nanoinformatics, various fundamental issues emerge. These include, for
example, the large number of different computer applications that have already arisen to deal
with the various areas and subjects of nanomedicine and nanotechnology, the large number of
papers already indexed in nanotechnology and nanotechnology bibliographic databases, the
number of companies and nanotechnologists working on nanotechnological and nanomedical
problems, nanomedicine medical aspirations, promising breakthrough development in different
areas of biomedicine, and the economic and ethical implications of this report. The growing
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value of the field will lead professionals in the region to develop the requisite nanoinformatics
approaches, in one way or another. These will pose a large range of new obstacles to science.
REFERENCES
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vol. 5, pp. 1909-1917, 2006.
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IMPORTANCE OF MICROBES IN PHARMACEUTICALS
Dr.K.Shoba* and J. Buenefa Shalom Trephosa
Department of Biochemistry, D.K.M. College for Women (Autonomous), Vellore,
Tamilnadu, India.
Email ID* - danishoba@gmail.com,
ABSTRACT
The study of microorganisms associated with the manufacture of pharmaceuticals, e.g.
reducing the number of microorganisms in the process environment, excluding microorganisms
and microbial bi-products such as exotoxin and endotoxin from water and other starting
materials, and ensuring that the finished pharmaceutical product is sterile. The production of
antibiotics is the most significant contribution of microbiology to the pharmaceutical industry.
Originally, all antibiotics were microbial products.Vaccines are also a very significant
contribution of microbiology to the production of medicines. In general, the manufacture of
vaccines against bacterial diseases involves the development of large quantities of bacteria.
Steroids can also be derived from microorganisms. Other aspects of pharmaceutical
microbiology include research and production of anti-infectious agents, the use of
microorganisms to detect mutagenic and carcinogenic activity in prospective medicines, and the
use of microorganisms, in the manufacture of pharmaceutical products like insulin and human
growth hormone.
KEYWORDS: Pharmaceuticals, Microbes, Drugs, Vaccines,Steroids, Hormones.
INTRODUCTION
Microbiological principles, procedures and techniques are applied to pharmaceutical
operations; the topic is then referred to as 'pharmaceutical microbiology.' It can be described as
the study of microorganisms that are important to the development of antibiotics, enzymes,
vitamins, vaccines and other pharmaceutical products; it also includes the study of
microorganisms that trigger pharmaceutical contamination and the degradation, deterioration and
spoilage of pharmaceutical raw materials and finished product.
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Pharmaceutical microbiology typically offers knowledge and understanding of the

importance of the existence of microbes, yeasts, molds, viruses and toxins in pharmaceutical raw
materials, intermediates, products and pharmaceutical production environments, as well as the
microbiological regulation of pharmaceutical products, manufacturing environments and
humans.
One of the main goals of pharmaceutical microbiology is to ensure the protection and
effectiveness of pharmaceutical products. It includes processes such as the validation of
disinfectants, assessment of the efficacy of disinfectants in suspension, on surfaces and through
field trials. Pharmaceutical microbiology provides procedures and techniques related to clean-
room process and assurance, aseptic and controlled environments to avoid potential microbial
contamination, and incorporate risk assessment and realistic contamination management
techniques.
Since the advent of this applied area of microbiology, pharmaceutical microbiology has
developed and grown greatly over the years to include numerous other aspects, such as research
and development of new anti-infective agents, the use of microorganisms to detect mutagenic
and carcinogenic potential in medicines, and the use of microorganisms in the manufacture of
insulin and human growth hormone.
ROLE OF MARINE MICROBES IN PHARMACEUTICAL DEVELOPMENT

Marine microbes have received increasing attention as a source of bioactive metabolites
and give a unique opportunity both to increase the number of marine natural products in clinical
trials and to accelerate their growth. This review focuses primarily on those molecules currently
in the therapeutic pipeline that are identified or highly likely to be generated by bacteria on the
basis of enhanced circumstantial evidence. an example of how compounds from harmful algal
blooms can produce lead, both for environmental change measurement tools and for
pharmaceutical production. An example of the carlotoxin class of compounds isolated from the
dinoflagellate Karlodinium veneficum shows a major environmental effect in the form of large
fish kills, but also offers opportunities for the construction of new cancer control molecules.and
serum cholesterol assisted by tools associated with rational drug design.
IMPORTANCE OF MICROBES IN NATURAL PRODUCTS
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Microbes are the leading manufacturers of valuable natural products. Natural products
made from bacteria and plants make excellent medicines. Major portions of the microbial
genomes are dedicated to the development of these valuable secondary metabolites. A single
microbe may make a range of secondary metabolites up to 50 compounds. The most valuable
items include antibiotics, anticancer agents, immunosuppressant,However, drugs have been
commercialized for many other uses, e.g. antivirals, anthelmintic, enzyme inhibitors,
nutraceuticals, polymers, surfactants, bio-herbicides and vaccines.
Unfortunately, owing to a decline in natural product development activities, drug

discovery has declined over the last 20 years.The reasons for this include high costs for clinical
trials, too short a timeframe before the drugs became generic, challenges in detecting antibiotics
against resistant species, and short treatment periods for patients for products such as antibiotics.
Despite these challenges, new drug discovery technologies have advanced, e.g., combination
chemistry of natural product scaffolding, biodiversity discoveries, genome mining, and systems
biology.The government's extension of time until goods became generic will be of great benefit.
MICROBES USED AS SOURCE OF ANTITUMOR DRUGS

Microbial metabolites are among the most effective chemotherapy agents for cancer. They
began to appear around 1940 with the discovery of actinomycin, and since then several
anticancer compounds have been isolated from natural sources. More than 60 per cent of the
existing compounds with antineoplastic activity were initially isolated as natural products or are
derived from them.Actinomycin D, anthracyclines (daunorubicin, doxorubicin, epirubicin,
pyrubicin and valrubicin), bleomycin, mitosenes (mitomycin C), anthracenones (mithramycin,
streptozotocin and pentostatin), enediynes (calcheamycin), taxols and epothilones are among the
authorized items.
MICROBES USED AS ENZYME INHIBITORS

Enzyme inhibitors have received increasing attention as useful tools, not only for the study
of enzyme structures and reaction mechanisms but also for potential utilization in medicine and
agriculture. Several enzyme inhibitors with various industrial uses have been isolated from
microbes. The most important ones are clavulanic acid, an inhibitor of β-lactamases. Some of the
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important targets for other inhibitors are glucosidases, amylase, lipase, protease and Xanthine
Oxidase (XO)
Amylase inhibitors are also known as starch blockers because they contain substances that
prevent the body from consuming dietary starches. Inhibitors can also be useful for weight
reduction, as some versions of amylase inhibitors have a capacity to reduce the absorption of
carbohydrates in humans. The use of amylase inhibitors to treat rumen acidosis has also been
confirmed.Examples of microbial amylase inhibitors are pain from Streptomyces corchorushii
culture filtrates and TAI-A, TAI-B, oligosaccharide compounds from Streptomyces calvus TM-
521 culture. Lipstatin is a pancreatic lipase inhibitor developed by Streptomyces toxytricini used
to regulate obesity and diabetes. It interferes with the absorption of fat in the gastrointestinal
tract. The commercial name is tetrahydrolipstatin, also known as orlistat.
MICROBES AS A SOURCE OF ANTI-FUNGAL

There is an increased need for new medicines that contributes to the treatment of diseases
in humans, animals and plants. A drastic example of this is the need for novel and more effective
antibiotics to fight multidrug-resistant microbial pathogens. Natural products are a significant
source of licensed drugs and still play an important role in the supply of chemical diversity,
considering the lack of interest on the part of large pharmaceutical firms.
New methods need to be placed in place to minimize the chances of the rediscovery of
tens of thousands of recognized natural products. The large number and variety of
chemotherapeutic agents derived from microbial natural products have contributed significantly
to the improvement of human health. However, only a small number of antifungal agents
(polyenes and azoles plus newly introduced caspofungin acetate) are currently available for the
treatment of life-threatening fungal infections.
MICROBES USED FOR TREATMENT AND PREVENTION OF ANEMIA

The traditional method for producing folic acid through chemical synthesis is known, where
the raw materials used are expensive and the yield of the final product is low. There are certain
folic acid-producing bacteria and yeast that accumulate high concentrations of folic acid in the
medium and may also be grown in whey or milk plasma.The numerous bacteria known that
generate and enhance the uptake of folic acid are Lactococcus lactis sub sp. Kremoris, L. It's
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lactis sub sp. Lactis, Bifidobacterium teenageis, B. Pseudocatenulatum; yeasts such as Candida
famata, C. Gilliermondii, C. Glabrate, Yarrowia lipolytica, Saccharomyces cerevisiae, Pichia
glucozyma, and Yarrowia lipolytica. The bacteria producing vitamin B12 are Pseudomonas
denitrificans and Propionibacterium shermanii.
Probiotic bacteria can also be used to avoid megaloblastic anaemia. It includes lactic acid-
fermented foods that increase the absorption of iron by optimizing the pH in the digestive tract,
stimulates enzyme phytases, and produces organic acids and other digestive enzymes.
The probiotic bacteria Lactobacillus plantarum decreases the harmful effects of the antibiotic
on colonic fermentation. Several medicines have recently been developed using genetically
modified bacteria or fungi that synthesize the medicine in giant bioreactors. Erythropoietin can
be man-made in bacterial bioreactors and used to treat anaemia, but treatment involves regular,
even daily injections. Researchers have purified human blood specializedcells that usually repair
the lining of blood vessels. These cells have been genetically modified to produce erythropoietin.
The cells were then combined with mesenchymal stem cells capable of forming blood vessels.
This mixture was then injected under the skin of mice that had been anaemic either by radiation
(as is often the case in chemotherapy patients) or by loss of kidney tissue.
MICROBES FOR TREATMENT OF ALLERGIES

Increased incidence of atopic diseases,atopic dermatitis, allergic rhinitis and asthma have
been documented. Evidence indicates that particular strains of probiotics have an impact on
inflammatory processes as shown by the reduction of some local and systemic immune markers,
and these activities may be mediated by GALT, one of the three intestinal lines of
protection..Probiotics can affect the development of inflammatory cells and the accessibility of
allergens, normalize the intestinal micro biota, affect the function of the intestinal barrier and
help regulate the secretion of inflammatory mediators.
CONCLUSION
Future perspectives will suggest that there is continuous growth in the number of new
microbial metabolites; however, qualitative progress is far more essential. This reflects important
functional effects both in human therapy and in agriculture. Much of the world's biodiversity
remains unexplored and modern high-speed approaches enable it to be successfully exploited.
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Cloning and genetic engineering provide alternate methods and the prospect of integrating
suitable biosynthetic pathways from non-culturable strains into appropriate hosts. Natural
products, in general, from microbes, may be expected to play an important role in the ongoing
transition from empiric screening to real logical drug design.
REFERENCES
1. Alarcon J, Aguila S, Arancibia-Avila P, Fuentes O, Zamorano-Ponce E, Hernandez M

(2003) Production and purification of statins from Pleurotusostreatus (Basidiomycetes)
strains. Z Naturforsch 58:62–64
2. Alberts AW, Chen J, Kuron G, Hunt V, Huff J, Hoffman C, Rothrock J, Lopez M, Joshua
H, Harris E, Patchett A, Monaghan R, Currie S, Stapley E, Albers-Schonberg G, Hensens
O, Hirshfield J, Hoogsteen K, Liesch J, Springer J (1980) Mevinolin. A highly potent
competitive inhibitor of hydroxymethylglutaryl-coenzyme Areductase and a cholesterol-
lowering agent. ProcNatlAcadSci USA 77:3957–3961
3. Bermudez-Humaran LG, Aubry C, Motta JP, et al.
Engineering lactococci and lactobacilli for human health. CurrOpinMicrobiol.
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4. Dufosse L. Pigments. Micro EncylMicrobiol. 2009;4:457‒471.
5. Gupta C, Prakash D, Garg AP, et al. Nutraceuticals from Microbes. In: Prakash D,
Sharma G, editors. Phytochemicals of nutraceutical importance. UK: CABI International
Publishers; 2014. p. 79‒102.
6. Newman DJ, Cragg GM. Natural products as sources of new drugs over the last
25years. J Nat Prod. 2007;70(3):461‒477.
7. Saghee M, Sandle T, Tidswell E (editors) (2011). Microbiology and Sterility Assurance
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8190646741.
8. Sandle, T. &Saghee, M.R. (2013). Cleanroom Management in Pharmaceuticals and
Healthcare.Passfield, UK: Euromed Communications
9. Sandle, T. (2012). The CDC Handbook: A Guide to Cleaning and Disinfecting
Cleanrooms. Surrey, UK: Grosvenor House Publishing. pp. 1–30. ISBN 978-
1781487686.
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10. https://lab-training.com/2015/03/07/how-microbiology-contributes-to-the-
pharmaceutical-
industry/#:~:text=Microorganisms%20are%20used%20in%20the,a%20small%20number
%20of%20microorganisms.
11. https://pharmaceutical-microbiology.imedpub.com/pharmaceutical-microbiology-an-
applied-branch-of-microbiology.php?aid=7860
12. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2994104/
13. https://link.springer.com/article/10.1007/s10295-013-1325-z
14. https://medcraveonline.com/JMEN/natural-useful-therapeutic-products-from-
microbes.html
15. https://medcraveonline.com/JMEN/natural-useful-therapeutic-products-from-
microbes.html
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ACQUISTION OF ELECTROSPIROGRAM
Chetana Krishnan
B.E Biomedical Engineeing, SSN College of Engineering
E.Mail ID - Chetanakrish20@Gmail.Com
ABSTRACT
Compliance is one of the important parameters that is used in invasive patient monitoring
systems to diagnose major lung diseases which cannot be tracked from scans and blood tests.
Unfortunately there are some limits like it cannot be used for continuous monitoring purposes
due to large power consumption and heavy weight. Hence it is important to develop a non
invasive and lightweight compliance meter which can be used for continuous monitoring
purposes. With respect to COVID pandemic, there is no proper tool to diagnose sars directly.
Hence measuring compliance can help us know the presence of sars as well as intensity through
which the virus has affected the lungs. This paper proposes the design and implementation of a
low cost, light weight and noninvasive compliance meter. Two sets of assumptions are taken to
implement the same. The first one is to measure the compliance of the healthy person and the
succeeding one is to measure the compliance of the abnormal patient. The healthy wave will be
made in the software and the abnormal wave will be compared with the ideal values to give the
final output. Since a comparative method is followed, the accuracy and efficiency of the meter is
increased. Also self calibration is achieved. This prevents replacement of the product then and
then and there. The data thus collected will be transferred to the nurse station for data
visualization. This promotes an alert system and reduces human labor.
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KEYWORDS: Compliance, Simulation, Pressure Vs Volume, Bolt, Comparison, Compliance
Degrees.
I.INTRODUCTION
There is an immense improvement in the ventilator systems used mainly in intensive care units
and critical care units. Compliance consists of two types - static and dynamic. Static compliance
refers to the stable compliance measurement whereas dynamic refers to unstable compliance
measurement. The former is used in tracking the lung status in case of TB and other noncausal
disorders. The latter is used to diagnose pathogen related lung disorders. But due to expenses and
high power consumption, continuous monitoring becomes a challenging task. According to
MDR regulations for medical devices, validation becomes a major challenge in case we try to
reduce the size of the device. But during an outbreak like COVID, non invasive and portable
compliance meters are much needed ones. Hence accuracy testing and clinical trials become a
restriction. The proposed meter can be used in non invasive ventilators or in any roadside booth
as the size is very much reduced. Lung monitoring of those patients post COVID becomes a
tedious job as no tool is developed as far as now except for temperature measurement.
The rest of the paper is organised as follows, Section 2 contains the related works. Section 3 is
presented with comparative technicalities of traditional and proposed compliance meters. Section
4 contains the simulation results. Section 5 is dealt with the accuracy testing and clinical trials.
Section 6 is dealt with future work and enhancement followed by conclusion.
II. RELATED WORKS
Amy D. Droitcour, [1] worked on a clinical non invasive respiratory management system for
patients who are in need of artificial respirator. She made a comparative study with 56 patients
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who had similar conditions but of different age groups. Hence she figured out that the same
system cannot be used for all the age groups. This was one of the major problems in wearable
devices.
Haipeng Liu, [2] tried to develop the traditional respiratory device system making it eligible for
all the age groups. He extracted the RR using other physiological signals from the human body.
After several clinical trials, he came to know that extracting other signals required massive
medical equipment and patient’s cooperation as his method was invasive in nature.
Carlo Massaroni, [3] implemented contact based respiratory management systems which
involved oxygen electrodes and chemical electrodes. The main principle behind his system was
the transfer of gaseous molecules from one electrode to another givind a difference in pressure.
This difference will be synced with the person’s breathing pressure and hence the RR can be
extracted. This method was limited because of the size and accuracy of the device mechanism.
Rasa Izadnegahdar, [4] developed the first non invasive wearable respiratory measuring device
which can measure the parameters like lung capacity, tidal volume and reserve volume of the
lungs. She took a systematic approach towards the existing tools available and made a device to
overcome all the limits. But the model was expensive as it used high level inbuilt and internal
sensors which put a halt to continuous monitoring.
Ian Smith, [5] developed a thermal based respiratory system which can measure the RR of the
person. The major drawback with the system was that the imaging part of the device must be
replaced after every use and it was quite an expensive and time consuming process wholly.
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From the above analysis and references, we came to a conclusion that there is no wearable
tool which can measure the compliance degree of the patient non invasively, light weight, cost
effectively. There is no proper alert system installed with the device and cannot be promoted for
the sake of continuous monitoring systems. The main aim of a diagnostic tool when it comes
with respect with COVID is that it should be alerted, non invasive and portable and cost
effective.
III. COMPARATIVE TECHNICALITIES OF TRADITIONAL AND PROPOSED
COMPLIANCE METERS
A. Traditional Compliance Meter
The traditional compliance meter consists of an external spirometer connected to the
spirometer wall. The spirometer wall is connected with a mouthpiece through which the patient
will be asked to inhale. The pressure thus exerted will be measured and the resulting tidal
volume will be produced. Once the device is used, it cannot be used by the other person as the
mouthpiece alone cannot be replaced due to design. Hence the whole device must be replaced for
making it reusable. The pressure measurer loses its calibration over a period of time and hence
accuracy cannot be achieved over a period of time.
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Fig 1: Traditional Compliance Meter This meter is not suitable for ventilator diagnostic purposes
due to its large size and heavy power consumption.
B. Proposed Compliance meter
The proposed compliance meter has ideal wave inbuilt in it. The ideal wave is measured by
using Masco’s Law. This step is followed by the measurement of compliance degree in the
patient using sensor operation and the change in volume. The results are compared with the ideal
characteristics and the output is produced. Also since the device works comparatively, self
calibration is achieved every time the output is compared with the ideal waves.
Fig 2: Proposed Product Package
IV. SIMULATION RESULTS
A. Ideal Wave Measurement
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As part of measuring the ideal wave, an NTC thermistor will be fixed inside a nebulizer mask.
Three thermistors are used here in order to achieve clinical accuracy. Once the patient breathes
through the mask, the temperature changes during the inspiration is converted into pressure using
a thermistor. This change is processed using Masco’s law and the final result is given as pressure
Vs volume graph.
Fig 3 : Nebulizer mask with thermistor fixed
Fig 4 : Ideal wave. X axis: Time (s), Y axis: Compliance Degree (no unit)
B. Measurement in Patient
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For this case, we have considered a dummy model with a bottle filled with a known amount
of water which has a balloon blown with a known amount of air. A clinical volume sensor will
be placed inside the balloon and a clinical pressure sensor will be placed in the bottle neck. The
volume and pressure sensor are connected via an arduino hub library wirelessly. A syringe will
be placed on the side of the bottle. Its purpose is to inject and extract water from it. This is
analogous to person inhaling and exhaling. Suitable algorithm is done and the graph for pressure
Vs volume is obtained by changing the volume of water via injecting and extracting water from
the bottle. Here the air volume inside the balloon will be constant throughout which is analogous
to reserve volume in the lungs.
Fig 5: Dummy Model
For every change in the volume given as input, the graph is produced between pressure and
volume which is very much different from the ideal wave. Along with the sensor, an indicator
will be placed. If there is red light, it means that the patient’s breathing is not active i.e it's
ISBN: 978-81-948129-1-3 Page 131

stopped. If there is orange light, it means that the values are varied from the critical and threshold
values. This promotes an alert system.
Fig 6 : Patient’s Compliance graph, X axis: Volume (ml), Y axis: Compliance Degree (no unit)
C. Data Visualization and Feasibility
Now that the data are collected, these can be transferred to the nurse station using IOT. A
bolt module can be used with the product to transfer the data immediately and continuously.
Thus the doctors can view the data at any time by visiting the dashboard in bolt. One single bolt
module can handle 56 such compliance metres and hence the product is feasible for large scale
marketing and consumption.
D. Threshold Values
Inspirational capacity: 3500 ml
Functional Residual Capacity : 2300 ml
Vital Capacity: 4300 ml
Total Lung Capacity : 5300 ml
Compliance Degree : 500 - 550
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Since the proposed product is very much effective in all criteria compared with the existing
product, the product is sustainable.
V. Accuracy Testing and Clinical Trials

A. Accuracy Testing
Inorder to check for the accuracy of the proposed product, a professional spiroscope was used
and tested with the patient. The results from the spiroscope and the proposed product was
compared and it was found that the accuracy was near cent percentage.
Fig 7: Professional Spiroscope

B. Clinical Testing:
The proposed product was tested with 3 people and appropriate results were obtained from
all of them. This product was readily recommended by the doctors of global hospital, Chennai.
VI. FUTURE WORK AND CONCLUSION
A. Future Work
The data collected so far is in the numerical form. This can be converted into binary form
using suitable converters. This can help in having a patient's record.
B. Regulation for medical devices
The product can be exposed for official clinical trials under MDR regulations to get the no
objection certificate. This can later be given for market segmentation
Thus a non invasive, light weight, cost effective, alerted, portable compliance meter was
implemented and tested for accuracy.
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REFERENCES
1. Amy D. Droitcour, “Non-Contact Respiratory Rate Measurement Validation for
Hospitalized Patients”, Conf Proc IEEE Eng Med Biol Soc. 2009; 2009: 4812–4815, doi:
10.1109/IEMBS.2009.5332635.
2. Haipeng Liu, “Recent development of respiratory rate measurement technologies”,
Published 2 August 2019, 2019 Institute of Physics and Engineering in Medicine,
Physiological Measurement, Volume 40, Number 7.
3. Carlo Massaroni, “Contact-Based Methods for Measuring Respiratory Rate”, 2019 Feb;
19(4): 908, Published online 2019 Feb 21. doi: 10.3390/s19040908.
4. Rasa Izadnegahdar, “A Systematic Review of Tools to Measure Respiratory Rate in
Order to Identify Childhood Pneumonia”, PubMed: 29474107, Received: November 12,
2017 Accepted: February 23, 2018.
5. Ian Smith, “Respiratory rate measurement: A comparison of methods”, January 2011
with 692 Reads, DOI: 10.12968/bjha.2011.5.1.18.
6. Sabyasachi Sircar, “A simple device for measuring static compliance of lung-thorax
combine”, 01 SEP 2015 https://doi.org/10.1152/advan.00026.2014.
7. Deyu Tang, “The embedded gas law and pulmonary compliance healthy human
simulated lung PID control system”, 2014 International Conference on Information
Science, Electronics and Electrical Engineering, 14714997.
Video Links: (output videos)
8. Block1:
https://www.youtube.com/watch?v=d2Tv_25A_Tk
9. Block 2:
https://www.youtube.com/watch?v=gU9PhaGIURs
10. Block 3:
https://www.youtube.com/watch?v=4s56laYACPo&feature=emb_logo
11. Accuracy:
https://www.youtube.com/watch?v=RS6W5Y0SX_Q&feature=emb_logo
12. Future work:
https://www.youtube.com/watch?v=GtKNsl4Qe5c&feature=youtu.be
ISBN: 978-81-948129-1-3 Page 134

SYNTHESIS OF SKIN PROTECTANT FABRIC BY COMPLEX MIXTURE OF FISH

COLLAGEN AND CELLULOSE NANO CRYSTALS FROM CASSAVA EXTRACT
V. Kaviya, Dr.Sr.G.S.Mary fabiola and V.Ilakkiya
Nirmala College for Women, Coimbatore
ABSTRACT:
From the last decade, there is a necessity to develop products for the skin for its assurance and
control from microbial pathogens. Many research works controlled to find a better solution for
the utilization of waste products into a beneficial one. Among the different kinds of waste
products, collagen and cellulose are having more effectiveness, which can be used in various
fields. Cellulose NanoCrystals (CNCs) have the incredible potential to be great nanomaterials for
synthesizing advanced materials. Because of their flexible nature, they are in the textile,
biomedical, and pharmaceutical industries. Extracted forms of collagen from fish skin, cellulose,
were used in the Nano level. Extract of collagen from Red snapper’s skin has the potential effect
and acts as an alternative source, which can increase the added value to the food and textile
industrial waste. A NanoCrystals of cellulose from cassava root has antimicrobial activity. The
properties of Collagen and CNCs have been qualitative to prove against skin pathogens. The
entire agents were loaded into cotton fabrics and evaluated for antimicrobial actions and
durability. This review is an eye-opening for the impact of bio-waste from aquaculture and plant
products to make a useful biomaterial as a Skin Protectant Fabrics.
KEYWORDS: Bio-Waste, Collagen, Cellulose NanoCrystals, skin, fabrics
INTRODUCTION:
Clothing comfort is one of the foremost imperative properties of fabric materials. It
suggests understanding the interrelationship between fiber material, fabric structure, and tactile
aspects of textile materials. Fabric performance in terms of better physiological and sensorial
comfort ought to be essential requirement materials that are using in contact with the skin (1).
The creation of keen fabric materials that can react to change of specific properties is the
foremost critical reason for the functionalization of materials. To plan useful materials for
stimuli-respond polymers can be utilized. Medical fabrics are one of the fast-growing segments
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of the global technical textile industry and one of the foremost important for growing divisions
within the specialized material showcase(2).
Skin is the primary defence boundary of human that has numerous imperative parts in
body assurance from outside intrusions and diseases; extreme skin disabilities would unfaithfully
influence the texture. The leadership challenge for skin caregivers is the emergence of
contamination from antibiotic-resistant microorganisms in traumatic wounds (3). Even though
numerous wide range antimicrobials are implicating for control of these safe strains, the
antimicrobial treatment is frequently not adequate to diminish wound bio burden microbes
beneath combat conditions (4). For potential overcoming the resistance issue, novel antibiotics
choices ought to be screen to treat wound contamination.
Collagen is from the significant extracellular protein lattice in dermal tissues, which plays
an energetically part in intervening cell migration and grip since of its specialized
acknowledgment interaction. Collagen-based dermal definitions are loyally connected for skin
regeneration, with assigning immunogenicity, tall biocompatibility, and have high porosity. (5)
Collagen may extricate from numerous living beings, the collagen created from sea-going life
forms is closing to human collagen, conjointly expand financial effect from the fishery
industry(6). Collagen productions from fish skin using the isolation process done on several
studies. Red snapper's Skin has been used as a collagen source through the acid-soluble method
in this study. The utilization of the acid-soluble methods has a few focal points, such as it
required several tools and time, deliver with a few waste and low production costs.
Amid the past decade, with the overexploitation and fatigue of fossil assets and genuine
natural disintegration, there's an increasing intrigued within the improvement of applications of
low-cost and renewable resources (7). Cellulose is the amplest biopolymer on the earth (8, 9) and
one of the broadly utilized polymers in nourishment, pharmaceutical, and bio-fuel sectors (10,
11). As of late, various thinks have centered on the segregation and generation of Nano cellulose
such as cellulose Nano Crystals (CNCs) acting as a bio based elective inside engineered tars
(12). Nano Crystals of cellulose are Normal, inexhaustible, renewable, bio-degradable, and tall in
quality and, making them alluring for creating bio-based, more economical item arrangements
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(13). Cellulose Nano Crystals (CNCs) can be Created from the characteristic cellulose by acid
hydrolysis or enzymatic strategy (14).
In recent days, there is an expanding inclination towards more valuable utilization of

biowaste into a beneficial one. Cassava (Manihot esculenta Crantz) is the most inexhaustible
starch crop delivered within the world, the third most vital nourishment source for tenants within
the tropical regions(15). The peels of cassava contribute to 15% of tubular cassava root weight
and the copious generation of this crop has brought about within the abdicate of a tremendous
sum of peels(16). The cellulose substance in cassava peel is quite promising at around 40-55% of
dry base peel(17, 18). Therefore, the cassava peel includes a potential as a source of cellulose.
Moreover, researchers have created textiles with antioxidant and antimicrobial properties
using the extracts of cellulose NanoCrystals and collagen. Antimicrobial property is important to
the cotton fabric using cellulose NanoCrystals of cassava peel extracts and fish collagen using
the pad-dry-cure method. This review work aims to report the new developments of medical
fabrics, focusing on the obtaining of bio-waste from cassava peel and fish skin containing
skincare substances, which are physically or chemically bound to the fiber surface.
MATERIALS AND METHODS:
FISH COLLAGEN EXTRACTION AND CHARACTERIZATION:

Red Snapper fish (Lutjanus campechanus) skins are obtaining from the aquaculture
waste. Then collected samples are being to keep at −20 °C until using, then thawing and
subjecting to the extraction process. (19) The entires steps are conducting at ≤4 °C, using double
distilled water for dissolving and washing. Briefly, the skin cut into ~2 cm2 pieces, the skin is a
wash, and non-collagenous proteins are deproteinizing using 0.1 N NaOH, rewashing and
lyophilizing. Collagen has to extract twice using immersion in 0.5 M acetic acid for 48 h,
filtration, and salting out with 1.0 M of NaCl. The precipitation of acid-soluble collagen was
collecting by centrifugation at 20.000 ×g for 45 min pellets are dissolving in acetic acid (0.5 M)
and dialyzing in diluted acetic acid (0.1 M) and Distilled water, collecting and lyophilizing. Then
the collagen is characterizing by SDS-polyacrylamide based gel electrophoresis (20) and by
spectroscopy (Fourier transforms infrared - FTIR) (21)
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Isolation of cellulose:
Cellulose Nano Crystals (CNC) are obtaining by sulfuric acid hydrolysis according to the
reported method (22). The pre-treating CPF powder, 1 g, is slowly adding to H2SO4 solution (63
wt %), 30 mL, under vigorously stirring at room temperature for 1h. After that, the hydrolysis is
quenching by adding ice water, 300 mL, into the mixture. The resultant mixture is first
centrifuging at 1000 rpm for 10 min to remove large particles, and then centrifuging at 11,000
rpm for 15 min to obtain cellulose Nano Crystals. The obtained cellulose Nano Crystals was
washing and centrifuging repeatedly for3 times before dialysis against distilled water for2 days.
Then they form cellulose Nano crystals is processing by the ultrasonic processor to suspension
better before further application.
Antimicrobial assays:
The growth inhibition zones (IZ) appearance, after well diffusion assay, is measuring as
an indicator for antimicrobial activity from Collagen (23) and cellulose are Diluting the solutions
from the extracting compounds, with a concentration of 1% acetic acid solution. 100 μL from
each microbial cell suspension (~2 × 106 CFU) is spreading into appropriate agar media, then
wells of 6 mm are made in agar and impregnating with 50 μL from diluting extracts. Plates (in
triplicates) are incubating at 37 °C for 18 h, and appearing IZs diameters' means are calculated.
Diluted acetic acid (1%) is used as a negative control.
APPLICATION OF COLLAGEN:
Cosmetics:
Water-soluble fish collagen is right now on the market as a valuable skincare item having
a moisture-retaining function. Sun harm (outward maturing) and aging (intrinsic maturing)
causes collagen within the skin to deteriorate. Other than the fish source, the collagen is
obtaining from creature sources and plant subordinates that act like collagen (pseudo-collagen)
utilized as a cosmetic ingredient. Collagen is in beauty care product that can be used to create or
build effective skin with glowing nature.
As implant:
Collagen-based products are broadly utilizing as vehicles for the transportation of cleared
skin cells or drug carriers for skin substitution (24, 25). They are using in refined skin treatment.
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Product of human keratocytes and fibroblasts which will join to make collagen
glycosaminoglycan substrates, which were subsequently cross-linked, diminished the rate of
biodegradation, and encourage decreased the engraftment of skin substitutes.
APPLICATION OF COLLAGEN
TISSUE REPLACEMENT SCAFFOLD
COLLAGEN
COSMETICS DRUG DELIVERY
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APPLICATION OF CELLULOSE NANO CRYSTALS:
IN PAPER MAKING INDUSTRY:

Cellulose NanoCrystals are highly crystalline, which are molecular orientation with high
mechanical quality, surface area, and high amount of hydroxyl groups. They are common used in
papermaking industries. The increases of cellulose NanoCrystals along with or without cationic
polyacrylamide and cationic starch in deinked pulp improved the maintenance of fillers, fibers,
and quality properties (26). NanoCrystals of cellulose proved that they have emulsion stability.
NANO MEDICINE:
CNC subordinates have utilized in definite Nano medical applications, such as the
antibacterial operators, tissue substitutions, biosensors, and bio imaging operators. The groups of
CNC-based materials will use for Myoblasts (muscle cells) seem to react to the sub-monolayer
surfaces of tunicin CNCs with a high degree of orientation (27), and thus profoundly situated
multinuclear myotubes formed (28). CNCs were used to fortify poly (ε-caprolactone) (PCL)
nanofibers. Other than the changes in mechanical properties and the thermal stability, the
nontoxicity to human cells and secure impact on directing cellular organization made the
nanofibers mainly valuable for artificial tissues or organs (29).
FOOD INDUSTRY:
The products from cellulose can be used in various forms. In the food industry, it can be
used as nourishment added substances such as an emulsifier, thickener, foam stabilizer, etc.
Cellulose Nanocrystals can act as a stabilizer because these strong particles accumulate at the oil-
water interface to form Pickering emulsions. They also have a role in the food packing
technique. Appear of CNCs in mechanical properties and water vapour penetrability of
biodegradable Nano composite film. The addition of CNC expanded the malleable quality and
malleable modulus of the film and altogether diminished the Water Vapour Permeability films.
The expansion of cellulose Nano crystals moved forward not as it were the water vapour barrier
but to the oxygen boundary properties of the bio-based nourishment bundling materials.
ISBN: 978-81-948129-1-3 Page 140

APPLICATION OF CELLULOSE NANO CRYSTALS
PHARMACUETICAL PAPER&PULP INDUSTRY

INDUSTRY
ISBN: 978-81-948129-1-3 Page 141

CELLULOSE NANOCRYSTALS
FOOD INDUSTRY NANO MEDICINE
APPLICATIONS OF NANO CELLULOSE FOR TEXTILE:
Nano materials for next-generation are useful to produce great textures and electronic
materials. It could be a promising fabric for creating low fetched, which is fully recyclable and
adaptable. They have an alluring property such as light weights, firmness, non-toxicity,
straightforwardness, gas permeability, and moved forward mechanical properties. (30, 31). They
utilized to permit slow-release functionalities of anti-microbial in cleanliness material products.
The utilization of nanomaterials can permit improved recolor repellence, diminish inactively, and
progress electrical conductivity to filaments without compromising their consolation and
adaptability. Cellulose nanofibers are more absorbent than superabsorbent polymers (SAPs) and
biodegradable (32). More particularly, this article will appear to think about the Nano cellulose
treatment of fabrics.
CONCLUSION:
Derivatization of CNCs is an effective arrangement to overcome the destitute chemical

resistance and mechanical properties of CNCs, such as uniting the functionalized moieties to
endow them with the shape memory impact, healable impact, or other particular effects (33, 34).
The destitute chemical safe property and the restricted mechanical property moreover
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constrained the scope of applications. As examined in the application segment, the arrangements
of CNCs and CNCs-derived materials are the centres to encourage handling CNCs into high-
performance composites and crossover materials.
In this manner, the high-cost CNCs prefer to chase for “high-value targets” (35).
However, strongly hydrophilic property, accumulation, and degradation of CNCs should be
tended to amid the unused applications and have to assessed with extensive studies.
Cellulose, the foremost antiquated and vital characteristic polymer on the earth, revives
and draws in a lot of consideration within the modern frame of CNCs can be utilized as a novel
and advanced materials and matrix. Widespread applications of CNCs within the papermaking
industry, reinforcing filler for polymers, shape memory polymers, healable polymeric materials,
nourishment industry, a sedate carrier within the pharmaceutical industry, supporting network for
catalysts, and nanomedicine was summarized in this paper.
The properties of collagen are varied with fish species and different parts of fish as well
as living environments of fish such as tropical and sub-tropical warm-water. Increasing the
demand for collagen has been found in many applications in food industries as emulsifiers,
foaming agents, colloid stabilizers, hydrogels; in food packaging industries as biodegradable
edible film, micro-encapsulating agents; and in pharmaceutical, biomedical industries as
capsules, bioactive peptides were used. Recently many studies urged to identify the impact of the
collagen bioactive peptides on anti-cancer, anti-oxidant, anti-diabetic, antiaging, cholesterol-
lowering agents, anti-atherosclerosis activity. Therefore, many modern analytical methods have
developed to characterize the fish collagen and bioactive peptides properties from all new
sources. Due to the excellent bioavailability and natural- like incorporation into the host tissue,
fish collagen could be used as a suitable biopolymer in biomedical and pharmaceutical
applications. With the above states, the future study aimed to produce fabric materials using bio
waste like collagen and cellulose from plant and aquaculture products for the sustainable future
generation.
ISBN: 978-81-948129-1-3 Page 143

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7. J. Song, C. Chen, S. Zhu and M. Zhu et al., Nature, 2018, 554, 224–228.
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13. Eichhorn SJ (2011) Cellulose nanowiskers: promising materials for advanced
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ISBN: 978-81-948129-1-3 Page 146

APPLICATION OF NANOTECHNOLOGY IN CANCER BIOLOGY
Dr. K. Shoba* and Ms. K. Sathvika
Department of Biochemistry, D.K.M. College for women (Autonomous), Vellore,

Tamilnadu, India.
Email ID: danishoba@gmail.com
ABSTRACT:-
This review tells about Application of Nanotechnology and its particles in cancer
biology. Nanotechnology is the manipulation of matter on atomic, molecular and supramolecular
scales. Nanoparticles are essential components for the development of future energy systems.
The highly size-related properties of nanoparticles provide uncountable possibilities for
unexpected discoveries. The frequently unpredictable and unprecedented activity of
nanoparticles has tremendous potential for groundbreaking technical applications, but it also
presents great challenges for scientists. Highly controllable synthesis methods, more receptive
characterization techniques and ultimately, new models and hypotheses need to be established to
describe them. Nanoparticles have lots and lots of application and which one among them is
application of nanotechnology in medicine specifically in cancer biology. Cancer remains the
second leading threat to human survival in the world and was responsible for an anticipated 9.6
million deaths in 2018. The main problem is we can’t able to diagnose a cancer in the early stage
with the present traditional methods and so we can able to diagnose the cancer only in the later
stages! In the later stages, the victim have to go for radiation and chemotherapy. So, by using
nanotechnology the victim can go for early detection of cancer and protect the patient from
cancer death. Nanoparticles are used in diagnosis, therapy and for drug delivery. Many nano
materials such as Carbon dots, Quantum dots, Titanium di oxide, Iron are used in cancer
diagnosis. Nanoshells are used to target cancer cells, Albumin Nano particle are used to deliver
anti-cancer drugs and Dendrimers is also utilized as a drug nanocarriers. The text has been
organised in five sections: Introduction, nanoparticle synthesis, Application of nanoparticles in
cancer biology, diagnosis and therapy, and Future scope.
KEYWORDS:
Nanotechnology, Nanoparticles, Carbon dots, Quantum dots, Albumin nano particle
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INTRODUCTION:
Nanotechnology or nanotech in short is the technology that involves the manipulation of

matter on atomic, molecular, and supra molecular scales. This includes particles of a scale of 1 to
100 nanometers. The term “Nano” comes from Latin word meaning Dwarf and in scientific
community it refers to 1nanometer. 1nm = one millionth of a mm and 1nm = one billionth of a
mm.
1000 Years ago (middle ages) Gold nano particles of different sizes are used to produce
different colors in stained glass windows. The term “Nanotechnology” was used for the first time
by Richard Feynman in 1959 in his article “There is plenty of room left at the bottom”. Materials
and devices are built from the molecular components which assemble themselves chemically by
principles of molecular recognition called as Bottom- up and the nano objects are constructed
from larger entities called as Top- down. Nanotechnology may be able to develop many different
materials and devices for a wide variety of uses, such as medicine, electronics, biomaterials and
energy production. Applications of Nanotechnology in medicine field which plays a vital role in
cancer diagnosis, therapy, drug delivery and also as a carrier. Cancer is a generic term for a
group of more than 100 diseases that can affect any part of the body. One defining feature of
cancer is the rapid creation of abnormal cells which grow beyond their usual boundaries, and
which can invade adjoining parts of the body and spreads to other organs, a process referred to as
metastasis. Cancer remains the second leading threat to human survival in the world and was
responsible for an anticipated 9.6 million deaths in 2018. The biggest concern is that we can’t
detect cancer at an early stage with the present traditional methods, and so we could able to
diagnose the cancer only in the later stages. In the later stages we have to go for radiation and
chemotherapy. So, by using nanotechnology the victim can go for early detection of cancer and
we can protect the victim from cancer death. Nanomaterials such as Carbon dots, quantum dots
are used in diagnosing cancer and nano shells are used in cancer therapy, Albumin nanoparticles
are used in drug delivery.
SYNTHESIS OF NANOPARTICLE:
Nanoparticles can be generated using a variety of methods. Nano particles can be synthesized
by physical, chemical and biological methods. Physical methods include attrition, pyrolysis, RF
plasma, Thermal decomposition, pulsed laser methods, while in chemical methods, we will be
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using reducing agents which are toxic and in biological synthesis of nanoparticle, a green method
and chemical reducing agent are not used instead microbial enzymes or plant phytochemicals act
as a reducing agent. Although some strategies are bottom up, some are considered top down.
CARBON DOT SYNTHESIS:

Carbon dot can be synthesized in many ways. One of the method is to synthesize carbon dot is
using Chitosan as a carbon source. It is a naturally occurring polysaccharide, cationic, highly
simple, muco-adhesive, bio-compatible polymer and licenced by the U.S. FDA for tissue
engineering and delivery of medicines. Degradation of chitosan depends on the degree of
deacetylation and the availability of amino groups. PEG used as a passivating agent. As chitosan
and PEG 4000 are combined with water and sulphuric acid. This solvent combination is exposed
to microwave irradiation (supplied with a domestic microwave oven). This combination of
solutions is dehydrated. Due to the dehydration process, the surface is passivated with
multicoloured carbon dots. Carbon dots can be synthesized by this technique.
SILVER NANOPARTICLE SYNTHESIS:

Silver nanoparticles can also be synthesised in a variety of techniques. One way to synthesise
silver nanoparticles is by a chemical synthesis process. For synthesizing silver nanoparticle by
chemical methods, we need three important components:
1. Metal salt
2. Reducing agent
3. Capping agent
The reducing agent will reduce the metal salt into metal nano particles but these metal
nanoparticles are highly unstable. So, it will join and form agglomerations or aggregations. So, to
make nanoparticles stable, the capping agent or stabilizing agent must be added, this will prevent
the aggregation or agglomeration.
The procedure is as follows: 1ml of deionized water was taken in 1.5ml micro centrifuge tube,
to this add 1.67 microliter of Silver nitrate solution is of 0.1M. To this mixture immediately add
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1.33 microliter of freshly prepared 2mg/ml NaBH4 solution is added. Finally, SDS (Stabilizer) of
90 microliter is added. These contents are mixed vigorously.
OBSERVATION AND RESULT:

 The development of a yellow colour will be seen, which means nanoparticles have been
created.
 The reducing agent which plays an important role in formation of yellow in colour
suggesting the development of silver nanoparticles.
APPLICATION OF NANOPARTICLES IN CANCER BIOLOGY:

Early diagnosis is a crucial consideration for effective treatment in the fight against cancer.
However, early cancer diagnosis has been hampered by the fundamental shortcomings of
traditional cancer diagnostic techniques. Nanotechnology has a high precision, accuracy and
multiplexed measuring capability and has also been studied for the identification of extracellular
cancer biomarkers and cancer cells, as well as for in vivo imaging. This review summarizes the
new trends in cancer diagnosis, therapy and drug delivery system using nanotechnology.
NANOMATERIALS IN CANCER DIAGNOSIS:

Nano devices can make cancer tests faster and more efficient. So, in cancer diagnosis various
nano materials plays an important role. The main important nano material is ‘’QUANTUM
DOTS’’. Quantum dots are highly fluorescent, nanometer- size, with respect to size it will give
different colour and fluorescence. Mechanism involved: Quantum dots are binded with an
antibody, this will specifically go and bind to the cancer cells and when the crystals are
stimulated by UV light, each bead emits light that serves as a sort of spectral bar code, making
the critical, cancer associated DNA sequences visible. Quantum dots have excellent brightness
and photo stability. Since quantum dots are made up of heavy metals, the use of quantum dots
for human use requires a lot of research and clinical trials. Carbon dots are considered to be
fluorescent nano materials and it is used alternatively to conventional toxic metal based quantum
dots. These are biocompatible and eco-friendly. These carbon dots will play a major role in
cancer diagnosis in near future.
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NANOMATERIALS USED IN CANCER THERAPIES:

Nano shells are widely used in thermal ablation of cancer cells. Nano shells have metallic
outer layer and silica core. These nano shells can be targeted to cancer cells for their destruction.
Nanoshells can be introduced and it specifically binds only to cancer cells and when infrared
light is applied, it generates heat. As a result it destroys only the cancer cells and the healthy cells
are intact. Liver Cancer and kidney tumors are now being treated with thermal ablation.
USAGE OF ALBUMIN NANOPARTICLE FOR DELIVERY OF ANTI -CANCER

DRUGS:
Mechanism is as follows: Make albumin nano particle with the size of 100- 250nm. Active
drug is introduced into albumin. Active drug in nano particle is non-crystalline form amorphous.
Here we can load hydrophobic drugs into hydrophobic packets to these albumin nano particles.
Drugs used here is Paclitaxel or curcumin. We can control the drug release by simple cross
linking reaction. So that the drug is slowly released. Abraxane an example of NAB (Nano
particle Albumin Bound).
DENDRIMERS USED AS DRUG NANOCARRIRES:

Dendrimer is a artificial polymer. The advantage of dendrimer is we can load anticancer
drugs in several locations. For example: In the dendrimer cavity we can have the encapsulated
drug, on the surface we can have adsorbed drug.
CURRENT NANOTECHNOLOGY RESEARCH:

 The use of nanotechnology for diagnosis and treatment of cancer is largely still in the
development phase.
 However, there are already several nano carrier-based drugs on the market and many
more nano -based therapeutics in clinical trials.
 Monoclonal antibody nanoparticle complexes are under investigation for diagnosis as
well as targeted delivery of cancer therapy.
 Nanoparticle-based chemotherapeutic agents are already on the market, and several are in
clinical trials.
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CONCLUSION:
Over the last 150 years, numerous truly innovative and ground-breaking methods have been
created to cure cancer. Major advances have been made, mainly due to breakthroughs in both
bottom-up and top-down nanotechnology. With the aid of nanotechnology, early identification,
mitigation and care can be achieved with a high degree of precision and convenience, which is
effective and can be made safe. The use of nanotechnology in the development of nanocarriers
for drug delivery provides a great deal of optimism and excitement in the area of drug delivery
science.
REFERENCES:
1. Elham Abbasi, Sedigheh Fekri Aval, Abolfazl Akbarzadeh, Morteza Milani, Hamid
Tayefi Nasrabadi, Sang Woo Joo, Younes Hanifehpour, Kazem Nejati-Koshki &
Roghiyeh Pashaei-Asl, Dendrimers: synthesis, applications, and properties,Nanoscale
research letters,2014
2. Valizadeh, A., Mikaeili, H., Samiei, M. et al. Quantum dots: synthesis, bioapplications,
and toxicity. Nanoscale Res Lett 7, 480 (2012)
3. Pyrolysis. In: Wang Z., Liu Y., Zhang Z. (eds) Handbook of Nanophase and
Nanostructured Materials. Springer, Boston, MA (2003)
4. Zhang Y, Li M, Gao X, Chen Y, Liu T. Nanotechnology in cancer diagnosis: progress,
challenges and opportunities. J Hematol Oncol. 2019 Dec 17;12(1):137. doi:
10.1186/s13045-019-0833-3. PMID: 31847897; PMCID: PMC6918551.
5. Sharma, A., Das, J. Small molecules derived carbon dots: synthesis and applications in
sensing, catalysis, imaging, and biomedicine, J Nanobiotechnol 17, 92 (2019). ISSN:
1477-3155
6. Munawar A. Mohammed, Jaweria T. M. Syeda, Kishor M. Wasan, and Ellen K. Wasan*,
An Overview of Chitosan Nanoparticles and Its Application in Non-Parenteral Drug
Delivery, Articles from Pharmaceutics,(2017), PMCID: PMC5750659, PMID: 29156634
7. Qinghui Zeng, Dan Shao, Xu He, Zhongyuan Ren, Wenyu Ji, Chong-Xin Shan, Songnan
Qu, Jing Li, Li Chen, Qin Li, Carbon Dots as a Trackable Drug Delivery Carrier for
Localized Cancer Therapy in vivo, Journal of materials Chemistry B 4(30), (2016)
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8. Birendra Kumar1*, PR Yadav2, HC Goel1, M Moshahid A Rizvi1, Recent

Developments in Cancer Therapy by the use of Nanotechnology, Digest Journal of
Nanomaterials and Biostructures, Vol. 4, No. 1, March 2009, p. 1-12
9. Andrew D. Warren, Gabriel A. Kwong, David K. Wood, Kevin Y. Lin, and Sangeeta N.
Bhatia, Point-of-care diagnostics for noncommunicable diseases using synthetic urinary
biomarkers and paper microfluidics, National Academy of Sciences, February 2014, vol.
111 no. 10 3671-3676
10. Schiffman JD, Fisher PG, Gibbs P. Early detection of cancer: past, present, and future.
Am Soc Clin Oncol Educ Book. 2015:57-65. doi: 10.14694/EdBook_AM.2015.35.57.
PMID: 25993143
11. Feng Chen ,iD Kai Ma, iD Miriam Benezra, Li Zhang, Sarah M. Cheal, Evan Phillips,
Barney Yoo, Mohan Pauliah, Michael Overholtzer,‖⊥ Pat Zanzonico, Sonia Sequeira,
Mithat Gonen, Thomas Quinn, Ulrich Wiesner, iD and Michelle S. Bradbury ,iD, Cancer-
Targeting Ultrasmall Silica Nanoparticles for Clinical Translation: Physicochemical
Structure and Biological Property Correlations, Chem Mater. 2017 Oct 24; 29(20): 8766–
8779
12. DGandhali A Deshpande, 2015. "Cancer Nanotechnology- The Recent Developments in
the Cancer Therapy," Global Journal of Nanomedicine, Juniper Publishers Inc., vol. 1(1),
pages 1-6, May.
13. Ajay VasudeoRane, SabuThomas, Methods for Synthesis of Nanoparticles and
Fabrication of Nanocomposites, Elesvier publishers, (2018).
14. Madamsetty VS, Mukherjee A and Mukherjee S (2019) Recent Trends of the Bio-
Inspired Nanoparticles in Cancer Theranostics. Front. Pharmacol. 10:1264. doi:
10.3389/fphar.2019.01264
15. https://www.intechopen.com/books/application-of-nanotechnology-in-drug-
delivery/current-status-and-future-scope-for-nanomaterials-in-drug-delivery
16. https://nanohub.org/resources/22262/download/NACK_U3_Maeder_Nanoparticle_Synth
esis.pdf
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NANO DRUG DELIVERY SYSTEM FOR THE TREATMENT OF CARDIOVASCULAR

DISEASES
Aakancha Shaw, St. Xavier’s College (Kolkata)
ABSTRACT
We are already aware that cardiovascular diseases (CVDs) have become a serious threat to
human life and health. So, developing drugs for treating CVDs has become extremely important.
Although many drugs are available in the market , they are still facing issues and are far from
satisfactory due to poor water solubility, low biological efficacy and their non targeting nature.
Owing to the rapid development of nanoscience, nanotechnology offers a new solution to
treat CVDs. Nano drug delivery systems (NDDs) are a class of nanomaterials that provide a new
drug delivery system for the treatment of CVDs.
Now comes the question, How exactly do NDDs work?
The nano drug delivery system (NDDs) improves the safety and effectiveness of the drugs
by increasing the stability and water solubility of the drug, by prolonging the cycle time, by
increasing the uptake rate of target cells and by reducing degradation by enzymes.
The commonly found drug carriers include:
 Liposomes
 Polymer micellar Co- delivery system.
 Metal nanoparticles
 Non metallic nanoparticles
TARGETING STRATEGY
The metabolic time of nano-transporter drugs in the blood circulation may be prolonged as
compared to that of the conventional preparations. By regulating pH value, temperature, light,
the rate of nano-transporter drugs can be made to function for a longer duration. The 2 types of
strategies are:
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 PASSIVE TARGET TRANSFER
The enhanced vascular permeability and retention( EPR) effect can be used in various CVDs.
For example, the occurrence of AS is a chronic inflammatory process, where vascular
permeability is often increased. Vascular endothelial permeability provides an effective means
for NDDSs to enter the interior of the plaque from the lumen.
 ACTIVE TARGETED TRANSHIPMENT
Active targeting primarily refers to the functional modification of NDDSs with one or more
targets to allow the drug to reach the desired site, the targeted site. This can be executed by
introducing a functional group or active substance that specifically interacts with diseased tissues
or cells into the surface of the nano-drug carrier will enhance carriers targeting .
SUMMARY-
Hence, we can conclude that the nano-carrier is an efficient, specific and controllable
intracellular drug delivery method that has shown unique advantages in the diagnosis and
therapy of CVDs.
It can effectively solve the problems that remained unresolved by the conventional drugs. It
can enhance targeting, local drug delivery, controlled release, sustained release,and can reducing
toxicity while it is developing toward the multifunctional and integrated direction of diagnosis
and therapy. With the innovation of nanotechnology, the application of NDDSs would be
promoted, and new techniques and methods could be provided for effective clinical diagnosis
and therapy.
REFERENCES-
1. Alam T., Khan S., Gaba B., Haider M. F., Baboota S., Ali J. (2017). Nanocarriers as
treatment modalities for hypertension. Drug Deliv. 24, 358–369.
10.1080/10717544.2016.1255999
2. Alie N., Eldib M., Fayad Z. A., Mani V. (2015). Inflammation, atherosclerosis, and
coronary artery disease: PET/CT for the evaluation of atherosclerosis and
inflammation. Clin. Med. Insights Cardiol. 8, 13–21. 10.4137/CMC.S17063
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3. Anselmo A. C., Modery-Pawlowski C. L., Menegatti S., Kumar S., Vogus D. R., Tian L.
L., et al. . (2014). Platelet-like nanoparticles: mimicking shape, flexibility, and surface
biology of platelets to target vascular injuries. ACS Nano 8, 11243–11253.
10.1021/nn503732m
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GENE ANNOTATION AND PHYLOGENETIC ANALYSIS OF CARCININ PROTEIN

FROM CARCINUS MAENAS
Dr. K. Shoba and J. Buenefa Shalom Trephosa
Department of Biochemistry, D.K.M. College for women (Autonomous), Vellore,
Tamilnadu, India.
E.mai ID - danishoba@gmail.com
ABSTRACT:
Carcinus maenas, commonly known as the European green crab, shore crab, European
shore crab is one of the best-known and most successful marine invasive species. C. maenas is
an excellent osmoregulator; therefore it is a euryhaline species. The variety of natural and
anthropogenic mechanisms are responsible for the geographic spread of this crab, and its ability
to adapt physiologically to a broad range of salinities, temperatures and other environmental
factors has enabled successful establishment in these new habitats. The green crab Carcinus
maenas is an invader on the Atlantic coast in Canada and the USA. Carcinus maenas has 5
lateral spines on each side of carapace (shell) its native range, the green crab is mostly used as an
ingredient in soups and sauces. In addition Carcinin is a whey acidic protein (WAP) domain-
containing antimicrobial protein produced by the circulating Haemocytes of the shore crab,
Carcinus maenas serve as innate immunity. The nucleotide and protein sequence of
antimicrobial protein (Carcinin) from Carcinus maenas is retrieved from NCBI database in fasta
format. The highly expressed genes are selected on the basis of CAI values using, ACUA tool.
The functional unit analysis was done using GYM motif server and Helix-turn-helix, etc. Finally
the Phylogenetic and mutational studies of highly expressed sequence were done using Amigo,
Clustal W, Protein Variability Server, etc.
KEYWORDS:
Antimicrobial protein, Amigo tool, ACUA tool, GYM motif server, Helix – turn – helix,
Clustal W, Protein Variability Server, Carcinin, Carcinus maenas.
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INTRODUCTION:
Crabs are decapod crustaceans of the infra order Brachyura, which typically have a very
short projecting "tail" (abdomen) usually entirely hidden under the thorax. Crabs are generally
covered with a thick exoskeleton, composed primarily of highly mineralized chitin, and armed
with a single pair of chelae (claws). Crabs are found in all of the world's oceans, while many
crabs live in fresh water and on land, particularly in tropical regions. Crabs vary in size from
the pea crab, a few millimeters wide, to the Japanese spider crab, with a leg span of up to 4
meters (13 ft.). The infra order Brachyura contains 6,793 species in 93 families, as many as the
remainder of the Decapoda. The evolution of crabs is characterized by an increasingly robust
body, and a reduction in the abdomen. They have hard shell, 2 eyes, 8 legs, and 2 pinchers
(claws) - hence they are known as decapods. The 6 legs have a pretty obvious use,
movement. C. maenas and other crabs move sideways through the water with their 6 legs. Their
claws are used to fight off predators and attract mates. Both of those uses are very important if a
crab wants to stay alive and pass on its genes. In most decapods, the gonopores (sexual openings)
are found on the legs. However, since crabs use the first two pairs of pleopods (abdominal
appendages) for sperm transfer, this arrangement has changed. As the male abdomen evolved
into a narrower shape, the gonopores have moved towards the midline, away from the legs, and
onto the sternum. A similar change occurred, independently, with the female gonopores. The
movement of the female gonopore to the sternum defines the clade Eubrachyura, and the later
change in the position of the male gonopore defines the Thoracotremata.
Carcinus maenas Habitat: mud, sand, or rock substrates, submerged aquatic vegetation, and
emergent marsh, although soft bottoms.
Distribution: Carcinus maenas is found on shore (or) coastal parts of Europe and Africa, from
Iceland and Norway in the north, to Mauritania in the south, and the Baltic Sea in the east, from
high water to depths of 60 m in the sub littoral, but it is predominantly a shore and shallow water
species. It tolerates a wide range of salinities and is especially abundant in estuaries and salt
marshes.
It grows to a carapace up to 90 mm (0.090 m / 9.0cm) long and 210 mm (8.3 cm) width.
Female size range: 15-31mm, Male size range: 1-86mm. It is a highly fecund species that
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reaches maturity quickly, and females are reproductively mature after one to three years. C.
maenas is an iteroparous species and female crabs can mate multiple times during a breeding
season but probably only produce one clutch of eggs per year. A maximum clutch size ranges
from 185,000 to 200,000 fertilized eggs. The carapace has five short teeth along the rim behind
each eye, and three undulations between the eyes. The Colour of C. maenas varies greatly, from
green to brown, grey, or red. This variation has a genetic component, but is largely due to local
environmental factors. In particular, individuals who delay moulting become red-coloured rather
than green.
METHODOLOGY:
The sequence of Carcinin Protein in Carcinus maenas is retrieved from NCBI database. The
retrieved sequence is submitted to Amigo tool to know the annotation of gene. The retrieved
annotated genome sequence is submitted to Clustal W to know the multiple sequence alignment
of Carcinin protein and phylogenetic analysis is done using PhyML. The retrieved annotated
genome sequence is submitted to ACUA to know the gene expression values. The retrieved
annotated genome is submitted to GYM motif server and Helix-turn-helix to know the functional
analysis of Carinin protein. The retrieved annotated genome sequence is submitted to Dis-
EMBL, GLOBPLOT to know the structural analysis and disorder prediction of Carcinin protein.
The retrieved annotated genome sequence is submitted to Pcoils server to know the coiled-coil
region of Carcinin protein. The retrieved annotated genome sequence is submitted to RADAR to
know the signal sequence prediction. The retrieved annotated genome sequence is submitted to
know the mutational studies and epitope discovery of Carcinin protein using Protein Variability
Server.
RESULTS AND DISCUSIONS:
I.SEQUENCE RETRIVAL:
1) NCBI:
CARCININ-SHORE CRAB (Carcinus maenas):
PROTEIN SEQUENCE:
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>CAD20734.1 carcinin [Carcinus maenas]
MKVQTVAAVVVVAVVVAMTEARLFPPKDCKYWCKDNLGINYCCGQPGVTYPPFTKSH
LGRCPPVRDTCTG
VRTQLPTYCPHDGACQFRSKCCYDTCLKHHVCKTAEYPYY
NUCLEOTIDE SEQUENCE:
>AJ427538.1:79-411 Carcinus maenas mRNA for carcinin
ATGAAGGTGCAAACTGTAGCAGCCGTGGTGGTTGTGGCTGTGGTTGTGGCCATGACA
GAGGCAAGGTTAT
TCCCTCCGAAGGACTGTAAGTACTGGTGCAAAGACAACCTTGGAATAAACTACTGCT
GTGGCCAGCCAGG
AGTAACCTACCCACCTTTTACTAAAAGCCACTTGGGCAGGTGCCCTCCAGTCCGTGA
TACCTGTACTGGC
GTCAGGACACAGCTACCAACGTACTGTCCCCATGATGGTGCATGTCAGTTCAGAAGC
AAGTGCTGCTATG
ACACCTGCCTGAAGCACCACGTGTGCAAGACTGCCGAATATCCTTATTATTAG
The above results shows the protein and nucleotide sequence in fasta format of Carcinin protein
in Carcinus maenas.
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2) GENE ANNOTATION-Amigo:
The above results show the location of genes and the entire coding region of genes (Gene
annotation) of Carcinin protein.
NOTE:
 Carcinine The green Colour in Carcinine denotes the Carcinine trans membrane
transporter activity.
 CarT The blue Colour in Car T denotes the Carcinine transporter.
 Drosophila melanogaster The blue Colour in Drosophila melanogaster denotes the

Taxonomy browser.
3) PHYLOGENETIC TREE:
PHYML:
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The above results show the Multiple Sequence Alignment and Phylogenetic analysis of Carcinin
Protein.
4) GENE EXPRESSION ANALYSIS –ACUA Tool:
S.NO Name of the sequence CAI Value
1. Nucleotide sequence-shore crab 0.9489950832767493
2. Nucleotide sequence-drosophila melanogaster 0.9541980712225387
3. Nucleotide sequence-Blattella germanica 0.9542460600197507
With the database sequence of ACUA Tool, the above result shows the highly expressed
annotated nucleotide sequence with Carcinin protein.
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II. FUNCTIONAL ANALYSIS:
5) GYM MOTIF SERVER:
The above results show the functional analysis of Carcinin protein.
6)HELIX-TURN-HELIX:
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The above result shows the functional analysis of Carcinin protein.
NOTE:
 The red Colour in the amino acid denotes the functional region of Carcinin protein.
III.STRUCTURAL ANALYSIS:
7) RADAR-SIGNAL SEQUENCE PREDICTION:
IV.MUTATION STUDIES AND EPITOPE DISCOVERY:
8) PROTEIN VARIABILITY SERVER:
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NOTE:
 The red Colour in the graph denotes the Plot Variability.
CONCLUSION:
Carcinin is a whey acidic protein (WAP) domain-containing antimicrobial protein produced

by the circulating haemocytes of the shore crab, Carcinus maenas. The most commonly
expressed transcript of the protein was recombinantly expressed in bacterial fusion system to a
yield of ca. Antimicrobial peptides and proteins AMPs (Carcinin) is a diverse class of naturally
occurring molecules that are produced as a first line of defense. This protein can have broad
activity to directly kill bacteria, yeasts, fungi, viruses and even cancer cells. Antimicrobial
peptides often encompass a wide range of categories such as antifungal, antibacterial, and
antituberculosis peptides. Carcinin protein is mainly present within granulated Haemocytes,
gonads and eyestalks. A patty product is made from green crab mincemeat using restructuring
additives (transglutaminase, dried egg white, isolated soy protein). The protein and nucleotide
sequence from the genus “Carcinus” is retrieved from National Center for Biotechnology
Information in Fasta format. The retrieved sequence is submitted to databases having various
tools for antimicrobial peptides analysis and phylogenetic analysis like Amigo, Clustal W,
ACUA, etc. The Functional unit analysis of Carcinin protein is done using GYM motif server,
Helix-turn-Helix etc. Predictions of primary structure and protein disorders using Dis
EMBL,GLOBPLOT etc., Finally mutational studies and epitope discovery is analyzed using
Protein Variability Server etc., Further detailed analysis in other aspects is needed for promoting
antimicrobial protein. Carcinin is used as a therapeutic agent. In Future Carcinin protein will
have numerous applications in the field of medicine.
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Cyclometopa. Part II. The Families Portunidae, Cancridae and Corystidae. Journal of the
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3. C. E. DeRivera; G. M. Ruiz; A. H. Hines; P. Jivoff (2005). "Biotic resistance to invasion:

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walking crab, Libiniaemarginata (Decapoda, Brachyura, Majoidea)". Arthropod Structure
& Development. 38 (3): 179–194. doi:10.1016/j.asd.2008.12.002. PMID 19166968.
8. Hanks RW, 1961. Chemical control of the green crab, Carcinus maenas (L.). Proceeding
of the National Shellfisheries Association, 52:75-84.
9. Iain J. McGaw; Timothy C. Edgell; Michel J. Kaiser (2011). "Population demographics
of native and newly invasive populations of the green crab Carcinus
maenas" (PDF). Marine Ecology Progress Series. 430: 235–240. doi:10.3354/meps09037.
10. J. H. R. Goddard; M. E. Torchin; A. M. Kuris; K. D. Lafferty (2005). "Host specificity
of Sacculinacarcini, a potential biological control agent of introduced European green
crab Carcinus maenasin California" (PDF). Biological Invasions. 7 (6): 895–
912. doi:10.1007/s10530-003-2981-0.
11. K. D. Lafferty; K. D. Kuris (1996). "Biological control of marine pests". Ecology.
Ecological Society of America. 77 (7): 1989–
2000. doi:10.2307/2265695. JSTOR 2265695.
12. L. H. Sweat (August 21, 2009). "Pachygrapsustransversus". Smithsonian Institution.
Retrieved 2010-01-20.
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13. M. de Saint Laurent (1980). "Sur la classification et la phylogénie des

CrustacésDécapodesBrachyoures. II. HeterotremataetThoracotremataGuinot,
1977". Comptesrendus de l'Académie des sciences. t. 290: 1317–1320.
14. N. Cohen; J. T. Carlton; M. C. Fountain (1995). "Introduction, dispersal and potential
impacts of the green crab Carcinus maenasin San Francisco Bay, California". Marine
Biology. 122 (2): 225–237.
15. P. Chen; A.Y. Lin; J. McKittrick; M.A. Meyers (May 2008). "Structure and mechanical
properties of crab exoskeletons". ActaBiomaterialia. 4 (3): 587–
596. doi:10.1016/j.actbio.2007.12.010. PMID 18299257.
16. Queen's University, Belfast (October 10, 2007). "Declawing crabs may lead to their
death". Science Daily. Retrieved 2012-09-21.
17. R. Thresher; C. Proctor; G. Ruiz; R. Gurney; C. MacKinnon; W. Walton; L. Rodriguez;
N. Bax (2003). "Invasion dynamics of the European shore crab, Carcinus maenas, in
Australia" (PDF). Marine Biology. 142 (5): 867–876. doi:10.1007/s00227-003-1011-1.
Archived from the original (PDF) on 2007-06-12.
18. S. B. Yamada (2001). Global invader: the European Green Crab. Oregon Sea Grant.
p. 123. ISBN 1-881826-24-4
19. T. J. Carlton; A. N. Cohen (2003). "Episodic global dispersal in shallow water marine
organisms: the case history of the European shore crabs Carcinus
maenas and C. aestuarii". Journal of Biogeography. 30 (12): 1809–
1820. doi:10.1111/j.1365-2699.2003.00962.x.
20. Walker RL, 1984. Effects of density and sampling time on the growth of the hard clam,
Mercenaria mercenaria, planted in predator-free cages in coastal Georgia. Natilus,
98:114-119.
21. Yamada SB; Bublitz R; Hardege J; Dawkins M; Quaintance A; Suggs L, 2006. Summary
of Green Crab Pheromone Study: Feasibility of using sex pheromones in controlling the
European Green Crab.
22. Zeidler W, 1978. Note on the occurrence of the European shore crab Carcinus maenas
(Linn. 1758) in Australia. South Australian Naturalist, 53:11-12.
23. Shoba K. , Sowmiya S and Dr. Mazher sultana, World Journal of Pharmaceutical and
Life Sciences, ISSN 2454-2229, Vol. 3, Issue 1, 427-436. 10.
ISBN: 978-81-948129-1-3 Page 167
24. Shoba K. , Manjuladevi M , Dr. Mazher sultana, Biochemical analysis and gene
expression profiling on collagenase protein in fiddler crab, World journal of pharmacy
and pharmaceutical sciences, issn 2278 – 4357, volume 6, issue 3, 747-756
25. Shoba K., Hebsibah Elsie B. And Bavyasri S. Insilico Peptide Modeling Stuidies And
Structural Analysis On Ribulose -1, 5 Bisphosphate Carboxylase In GracilariaEdulis,
World Journal Of Pharmacy And Pharmaceutical Sciences, 2018,Volume 7, Issue 3,
1086-1095, Issn 2278 – 4357.
26. Shoba K., Kalpana K., Protein Modeling and Drug Docking Studies on Potential Protein
Target (E. coli–dosP) and Compound Aldehyde (Sumatriptan) using Bioinformatics
Tools. Research & Reviews: A Journal of Bioinformatics. 2018; 5(3): 9–18p.
27. Shoba. K, Hebsibah Elsie. B and Jayakumari. S. Sathya. R. (2018); Insilico Structural
Analysis and Drug Docking Stuidies On Ribulose -1, 5 Bisphosphate Carboxylase In
Gracilaria Edulis. International journal of advanced research. 6 (9). 159-165] (ISSN
2320-5407).
28. Shoba. K, Nithy.G and Deepa.L. (2018); Biochemical Analysis and Peptide Modeling of
Lysozyme in Indian Fenneropenaeusindicus shrimp species. International journal of
advanced research. 6 (9). 159-165] (ISSN 2320-5407).
29. Shoba.k and Dr. Mazher sultana, Three - dimensional structure and motif prediction
studies on collagenase protein in fiddler crab, International journal of novel trends in
pharmaceutical sciences, Issn: 2277 – 2782,volume 6, issue 4, pages 79 – 83.
30. Shoba.K, Hebsibahelsie.B, insilico homology modeling ofribulose-1, 5bisphosphate
carboxylase protein in gracilariaedulis, world journal of pharmacy and pharmaceutical
sciences, 2017,volume 6, issue 8, 396-406, issn 2278 – 4357.
31. Shoba.K, Lavanya.G, Identification Of De Novo Peptide And Motif Prediction On
Porphyria Protein (Hmbs) Using Insilico Tools, Universal Journal Of Pharmacy ,2018,
Volume 8, Issue 1, Issn2320-303x.
32. Shoba.K, Lavanya.G, Tertiary Structural Prediction And Drug Binding Studies On
Mutated Gene (Hmbs) In Human Porphyria,International Journal Of Novel Trends In
Pharmaceutical Science,2018,Issn 2277 -2782,Volume 8,Issue 1
ISBN: 978-81-948129-1-3 Page 168

33. Usha.K and Shoba.K, Functional analysis &pepptide structure modeling of sphyastatin
protein in scyllaparamamosain, International Journal of Current Science and Technology,
Volume 7, Issue, 01(A), pp. 666-669, January, 2019, ISSN: 2320-8090.
ISBN: 978-81-948129-1-3 Page 169

EVALUATION OF DRUG SUSCEPTIBILITY WITH ALAMAR BLUE IN

MYCOBACTERIUM TUBERCULOSIS
J. Albino Wins and M. Murugan
Department of Botany, Holy Cross College (Autonomous), Nagercoil – 4
(Affiliated to Manonmaniam Sundaranar University, Abishekapatti, Tirunelveli District)
*Department of BioMedical Sciences, Noorul Islam Centre for Higher Education, Kumaracoil,
Tamil Nadu.
E.mail Id:winsbt@gmail.com
ABSTRACT:
Tuberculosis is a chronic disease, which is posing a serious threat in all parts of the world.
Since front line drugs and second line drugs were adapted as antituberculosis drugs, there is a
serious threat of increasing incidence of drug resistant strains and it emerged all over the world.
Thus treatment of TB is an important event and without the information on susceptibility to
drugs which makes the treatment failure. Thus prompt diagnosis and appropriate therapy for the
detection of drug resistance is needed. Amomg various types of techniques, Alamar Blue assay is
an sensitive method in which results can be identified visually within few days.
KEYWORDS:
Tuberculosis, Alamar Blue assay, Drug resistance, Antibiotics.
INTRODUCTION:
Today, as it has been for cenutries, tuberculosis remains the leading cause of death in the
world. The disease is now the world’s foremost cause of death from the single infectious agent.
Mycobacterium tuberculosis is the sensitive index of a nation’s poverty. Currently, the disease is
chronic, slowly debilitating and pose a serious threat to the health in developing and developed
countries. Tuberculosis is a re-emergent problem in many industrialized countries 1.
ISBN: 978-81-948129-1-3 Page 170

Different drugs like Isoniazid, Rifampicin, Ethambutol, Streptomycin, Pyrazinamide,

Ethionamide, Amikacin, Kanamycin and Capreomycin has been used. Antibiotics are available
that effectively inhibit bacterial cellwall synthesis, protein synthesis and DNA replication 2.
Bacteria can resist antibiotics as a result of chromosomal mutation or inductive expression of a
latent chromosomal gene or by the exchange of genetic material through transformation,
transduction or conjugationby plasmids. Resistance can be transferred horizontally by plasmis or
by chromosomally located conjugative transposons that spread the resistance to other species,
but rather mutation in the target genes3.
The antimicrobial agents are rendered inactive by three major mechanisms like, inactivation
of the antibiotic by destruction or modification, prevention of the access to the target and
alteration of the antibiotic target site4.
Drug susceptibility tests were done to determine the proportion of bacilli in the test
population , which is resistant to the drugs being tested5. However, susceptibility tests should be
done before therapy is started. Treatment of tuberculosis without the benefit of susceptibility
information increases the risk of treatment failure and spread of resistant strains, and so various
techniques were evolved for sensitivity testing6.
METHODOLOGY:
In the present investigation, sputum samples were collected from the pulmonary tuberculosis
patients. Early morning samples were collected and stained by Zeihl - Neelson technique. The
sputum samples were processed by Modified Petroffs method and the sediment was inoculated in
Lowenstein Jensen medium for growth.
The isolates were analysed for their susceptibility to the antibiotics like rifampicin (RMP),
isoniazid (INH), ethambutol (EMB) and streptomycin (STM). The stock solutions were thawed
on ice and required volume of drug were added to 450µl of TB broth in 1.5ml sterile disposable
microcentrifuge tubes to get 0.5X, 1X and 2X critical concentrations. The final concentrations
were: RIF, 0.5, 1 and 2µg /ml; INH, 0.5, 1 and 2µg /ml; and STM, 0.5, 1 and 2µg /ml.
ISBN: 978-81-948129-1-3 Page 171

The isolates were tested with three different concentrations of 4 drugs. The tubes were
incubated at 370C without shaking for about 7 days. On the 7th day, 25 µl of Alamar blue
solution was added and checked for colour change.
RESULTS AND DISCUSSION:
The colorimetric MIC results for all clinical Mycobacterium tuberculosis isolates were
obtained by sufficient growth to determine drug susceptibility. So, Alamar Blue dye was added
to all the five sets of tubes and the colour change was seen in the corresponding drug resistant to
that isolate. A definite colour change from blue to pink coloured at critical concentration of the
drug indicates resistance to that particular drug. The respective colour change in all the tubes was
tabulated within 6 hours. All the 5 isolates showed colour change within 2 hours and hence the
dye was added to all the remaining control tubes.
All the isolates were susceptible to rifampicin in subcritical, critical and double critical
concentrations. All the 5 isolates were resistant to isoniazid in subcritical concentration and 1
shows susceptible in critical concentration and 2 shows resistance in double critical
concentrations. In case of Streptomycin, all the isolates were susceptible to all the critical
concentrations. In case of ethambutol, 3 isolates showed resistance in the subcritical, critical and
double critical concentrations.
Table 1: Results showing Alamar Blue Assay
Isolate Rifampicin Isoniazid Streptomycin Ethambutol

Number
0.5 1 2 0.2 0.4 0.8 1 2 4 2.5 5 10
189 B B B P P P B B B B B B
194 B B B P P B B B B P P P
195 B B B P P B B B B P P P
196 B B B P B B B B B P P P
197 B B B P P P B B B B B B
B – Blue : Sensitive
ISBN: 978-81-948129-1-3 Page 172
P – Pink: Resistant
The Alamar Blue assay offers a superior combination of rapidity and affordability. This allows
the selection of the critical concentration of each drug for use in differentiating susceptible,
partially resistant and fully resistant strains7. Considering their rapidity, their use of low
technology and their low cost, the use of ABA have the potential of becoming the methods of
choice for drug susceptibility testing of Mycobacterium tuberculosis isolates for much of the
world, where this disease is a major problem. It is important to determine the critical
concentration of each drug that correlates with clinical resistance8. Working out the exact
biochemical details of drug – drug target interaction acquires considerable attention in the era of
multi drug resistance TB, because only then will more rational structure and mechanism based
approaches to inhibitor design be possible. A concreted global effort is required to defeat TB
resurgence.
REFERENCES:
1. Ajay Kumar, R., K. Laiza Paul R. Indulakshmi, Y.K.Manju, K.Vinod Kumar,

P.Ayyappan, M.Joshi and Sathish Mundayoor. Analysis of drug susceptibility in
M.tuberculosis isolated from Thiruvananthapuram using Alamar Blue, 2001. Current
Science.80: 70 – 74.
2. Alcaide, F., Pftyffer, G.E and Telenti,A. 1997. Role of EMB in natural and acquired
resistance to EMB in Mycobacteria. J.Antimicrob Agents Chemother. 41: 2270 – 2273.
3. Barry, R., Bloom. Tuberculosis Pathogenesis, Protection and control, 1994. American
Society for Microbiology, Washington, DC.
4. Hinshaw, H.C., Feldman,W.H. 1945. Streptomycin in treatment of clinical Tuberculosis:

Preliminary Report. Proc Staphy Med Mayo Clinic. 20: 313 – 318.
5. Petroff, S.A. 1915. A new and rapid method for isolation and cultivation of tubercle
bacilli directly from sputum and faeces. Journal of Experimental Medicine. 21 – 38.
6. Snider, Jr.D.E., Roper, W.L. 1992. The new Tuberculosis. New Engl J Med. 1993, 329:
784 – 791.
ISBN: 978-81-948129-1-3 Page 173

7. WHO report on TB epidemic. Global TB programme. Geneva: The Organization,1994.

94 -177.
8. Yajko,D.M., J.J.Madej, M.V.Lancaster,C.A.Sanders, V.L.Cawthon, B.Gee,A.Babst and

W.K.Hadley. 1995. Colorimetric method for determining MICs of antimicrobial agents
for Mycobacterium tuberculosis. J.Clin Microbiol. 33: 2324 – 2327.
ISBN: 978-81-948129-1-3 Page 174

GREEN FABRICATION OF HIGH PERFORMANCE ZN-CO FILM THROUGH

SUPERCRITICAL CO2 ELECTRODEPOSITION PROCESS: ITS CORROSION
EVALUATION
Manickaraj Shobana Sebastin Mary, Pandiyarajan Sabarison , Muthuraja Kalpana devi,
Sheng-Tung Huang , Ho-Chiao Chuang
Department of Chemical Engineering and Biotechnology, National Taipei University of
Technology, Taipei-10608, Taiwan, ROC
Department of Mechanical Engineering, National Taipei University of Technology, Taipei-
10608, Taiwan, ROC
PG & Research Department of Chemistry Bishop Heber College, Tiruchirappalli, India.
*Corresponding author: Email: hchuang@mail.ntut.edu.tw, Phone：+886-2-2771-2171
Ext: 2076.
GRAPHICAL ABSTRACT
ABSTRACT
This work demonstrates the zinc – cobalt (Zn-Co) metal film properties prepared via
supercritical CO2 (SC-CO2) electrodeposition. The films were also prepared by conventional
electrodeposition for the comparison studies. All the prepared films properties were
characterized altogether by scanning electron microscope (SEM), elemental dispersive X-ray
spectroscopy (EDX), X-ray diffraction spectroscopy (XRD), atomic force microscope (AFM)
ISBN: 978-81-948129-1-3 Page 175
and electrochemical analysis. Based on the results, the film prepared through SC-CO 2
electrodeposition process shows better coating quality than conventional method. The effect is
attributable to the fact that the bath blended with SC-CO2 has low gas surface tension and high
fluid mass transfusion. This nature efficiently improves the coatings’ deposition rate in the blind
via, and further tends to make the coating surface compact and more homogeneous. The obtained
result showed that a popcorn-like structure was observed with reduced crystallite size and
effectively improved anti-corrosion performance. Thus, the proposed novel SC-CO 2
electrodeposition method improves the Zn-Co composite films’ surface and electrochemical
properties.
KEYWORDS: Electrodeposition; Zinc-Cobalt; Supercritical-CO 2;
ISBN: 978-81-948129-1-3 Page 176

A STUDY ON THERAPEUTIC POTENTIALS AND MOLECULAR DOCKING OF
PLANT EXTRACTS AGAINST ENVIRONMENTAL ISOLATES CAUSING URINARY
TRACT INFECTION
Venkat S, Jayasree A, Pavithra M*, Shoba G* and Sivaranjini A*
D.G. Vaishnav College, Chennai.
svenkatmsd@gmail.com, pavithram@dgvaishnavcollege.edu.in
*Corresponding Author: sivaranjinia@dgvaishnavcollege.edu.in
ABSTRACT
Plants produce a diverse range of bioactive molecules, making them a rich source of
different types of medicines. A regular and widespread use of herbs throughout the world has
increased serious concern over their quality, safety and efficacy. The core of the project is to
study the antimicrobial activity of Tinospora cordifolia, Annona squamosa and Punica granatum
extracts of three different solvents like Petroleum ether, Methanol and Chloroform that were
used against 25 microbial samples isolated from different sources and the antimicrobial activity
of plant extract were studied and the study also specializes with the anti-bacterial inhibition of
Urinary Tract Infection (UTI)- Escherichia coli and Klebsiella pneumonia clinical isolates
obtained from UTI infected patients. The antimicrobial activity was carried out by using agar
well diffusion method and Minimum inhibitory concentration. The methanol extract of all three
plants showed good antimicrobial activity against UTI causing organisms. Standard biochemical
test was carried out for the characterization of gram negative bacteria. Molecular interaction
analysis technique was used to predict the binding of Protein-ligand and thus plays a key role in
structure based drug design. Molecular interaction analysis of six different compounds against
protein Ibestrophin (virulent protein of Klebsiella pneumonia causing UTI) performed using
bioinformatics tool revealed that the plant extract of Annona squamosa can be used as a natural
alternative therapy to control UTI diseases and prevent health hazards. This might be an effective
solution for emerging drug resistance for chemical antimicrobial agents.
ISBN: 978-81-948129-1-3 Page 177

PHYTOCHEMICAL AND ANTIOXIDANT ANALYSIS OF MEDICINAL PLANTS AND
ITS ANTIBACTERIAL EFFECT ON BACTERIAL PATHOGENS
M.K. Arya, Sathish D, Jayasree A, Sivaranjini A* and Pavithra M*
D.G. Vaishnav College, Chennai
*Corresponding Author: sivaranjinia@dgvaishnavcollege.edu.in
The beneficial medicinal effects of plant materials are usually due to Secondary products.
Many secondary compounds from plants are known to contain diverse biological activities which
may include antibacterial, antifungal and anticancer activities. In the present study, the leaf
extract of plants such as Eclipta Prostrate, Phyllanthus niruri, Senna auriculata were extracted
and the secondary metabolites from the plants were extracted using the solvents like Chloroform,
Methanol and Water. Qualitative analysis of secondary metabolites was studied and the
screening of secondary metabolites against bacterial strains including Escherchia coli, Klebsiella
pneumoniae, Psuedomonas syringae, Psuedomonas viridiflava, and Streptococcus pneumonia
was carried out using Kirby Bauer Agar Well Diffusion method. Determination of Antioxidant
capacity of E. prostrata, S. auriculata and P. niruri by DPPH assay and Characterisation of E.
prostrata was carried out by FTIR. The extracts of E. prostrata were proved to be potentially
effective and can be used as the natural alternative therapy as it possess secondary metabolites
that act as good antimicrobial, antioxidant agent and prevent health hazards and drug resistance
problems of chemical antimicrobial agent applications.
ISBN: 978-81-948129-1-3 Page 178

NANOBIOREMEDIATION
Abirami S
Research Scholar, Environmental Science
Indira Gandhi National Open University, New Delhi
sgabirami6@gmail.com
Environmental degradation and ecological imbalance is the major issue of global concern
which is to be addressed in eco-friendly and a sustainable way. The emerging advancements of
nanotechnology can address the major environmental challenges substantially better. Apart from
their unique properties (size, surface, and inner properties), the nanoparticles can function as
solids or liquids in the air (aerosols), solids in liquids (suspensions), and as liquids in liquids
(emulsions). These exclusive characteristics of nanoparticles can be employed for the potential
environmental cleanup that provides a sustainable environment. The use of nanomaterials
synthesized from microbes, plants, and algae for the bioremediation of a polluted environment is
known as nanobioremediation.
Microorganisms work at the microlevel and are capable of converting the toxic pollutants
into a non-toxic form within a short period. When paired with nanoparticles the microorganisms
acquire the capability to function at the nanolevel and thus, due to the increase in the surface
area, a large number of pollutants can be treated at a time. Additionally, the surface plasmon
resonance property and the quantum effect of nanomaterials helps in the detection of toxic metals
and require less activation energy to carry out the process of bioremediation. Several studies
have been carried to analyze the potential of nanomaterials synthesized from microbes and plants
as nanobioremediaters.
To name a few, The study conducted by Subramaniyam et al (2016) on the synthesis of

nanoparticles using the Chlorella sps found in the brewery wastewater revealed that the total
nitrogen, phosphorous, and organic carbon were completely removed and utilized by Chlorella
sps for their growth and resulted in nanoparticle production. The gold nanoparticles synthesized
from Aspergillus sps are found to be effective for the degradation of 2-nitrophenol, 3-
ISBN: 978-81-948129-1-3 Page 179

nitrophenol, 4-nitrophenol, o-nitroaniline, and m-nitroaniline (aromatic pollutants) (Qu et al.,

2017). The release of industrial effluents contains many harmful dyes which are known to inhibit
the light penetration in the water bodies and thus affect the photosynthetic activity. The study
conducted by Aminuzzaman and coworkers, (2018) on photocatalytic activity of zinc oxide
nanoparticles synthesized from Garcinia mangostana fruit showed the promising effect on
degradation of malachite green dye. Similarly, the titanium dioxide nanoparticle synthesized
from Bacillus amyloliquefaciens and Ageratina altissima showed great photocatalytic activity
against the reactive red 31, methylene blue, alizarin red, crystal violet, and methyl orange (Khan
and Fulekar, 2016 and Ganesan et al., 2016). The gold and carbon nanodots synthesized from the
Allium cepa leaves are proved to be efficient for the detection of Hg2+ ions in water
(Venkateswarlu et al., 2019).
To conclude, the field of nanobioremediation is still at the budding stage and more
research is needed to access the degree of safety for the biological systems.
REFERENCES
1. Aminuzzaman, M., Ying, L. P., Goh, W. S., & Watanabe, A. (2018). Green synthesis of
zinc oxide nanoparticles using aqueous extract of Garcinia mangostana fruit pericarp and
their photocatalytic activity. Bulletin of Materials Science, 41(2), 50.
2. Ganesan, S., Babu, I. G., Mahendran, D., Arulselvi, P. I., Elangovan, N., Geetha, N., &
Venkatachalam, P. (2016). Green engineering of titanium dioxide nanoparticles using
Ageratina altissima (L.) King & HE Robines. medicinal plant aqueous leaf extracts for
enhanced photocatalytic activity. Ann. Phytomed., 5(2), 69-75.
3. Khan, R., & Fulekar, M. H. (2016). Biosynthesis of titanium dioxide nanoparticles using
Bacillus amyloliquefaciens culture and enhancement of its photocatalytic activity for the
degradation of a sulfonated textile dye Reactive Red 31. Journal of colloid and interface
science, 475, 184-191.
4. Qu, Y., Pei, X., Shen, W., Zhang, X., Wang, J., Zhang, Z., ... & Zhou, J. (2017).
Biosynthesis of gold nanoparticles by Aspergillum sp. WL-Au for degradation of
aromatic pollutants. Physica E: Low-dimensional Systems and Nanostructures, 88, 133-
141.
ISBN: 978-81-948129-1-3 Page 180

5. Subramaniyam, V., Subashchandrabose, S. R., Ganeshkumar, V., Thavamani, P., Chen,

Z., Naidu, R., & Megharaj, M. (2016). Cultivation of Chlorella on brewery wastewater
and nano-particle biosynthesis by its biomass. Bioresource technology, 211, 698-703.
6. Venkateswarlu, S., Govindaraju, S., Sangubotla, R., Kim, J., Lee, M. H., & Yun, K.
(2019). Biosynthesized highly stable Au/C nanodots: ideal probes for the selective and
sensitive detection of Hg2+ ions. Nanomaterials, 9(2), 245.
ISBN: 978-81-948129-1-3 Page 181

HIRSCHSPRUNG INFECTION, RELATED DISORDER, AND HEREDITARY

QUALITIES: A SURVEY
Shubham
Dyal Singh College, Karnal (132001) , Haryana , India
Mobile no.-8708075152
ABSTRACT
This paper mainly aims to the Hirschsprung infection (HSCR, aganglionic megacolon) is the
principle hereditary reason for practical intestinal block with a rate of 1/5000 live births. This
formative problem is a neurocristopathy and is described by the nonappearance of the enteric
ganglia along a variable length of the digestive tract. In the most recent many years, the
advancement of careful methodologies has significantly diminished mortality and horribleness,
which has permitted the rise of familial cases. HSCR had all the earmarks of being a
multifactorial mutation with low, sex subordinate penetrance and variable articulation as
indicated by the length of the aganglionic fragment, proposing the inclusion of at least one
gene(s) with low penetrance. Up until now, eight qualities have been discovered to be associated
with HSCR. This incessant inborn mutation currently remains as a model for hereditary problems
with complex examples of legacy.
KEYWORDS:- Hirschsprung infection (HSCR), hereditary, mutations, Genes, ganglia etc.
ISBN: 978-81-948129-1-3 Page 182

DEVELOPMENTAL HEREDITARY QUALITIES OF PLANT VARIATION
SHUBHAM
Mobile no.-8708075152
ABSTRACT
In this paper, we will discuss on Plants give special occasions to consider the robotic
premise and developmental cycles of variation to assorted ecological conditions. Corresponding
research center and field tests are significant for testing theories reflecting long haul biological
and developmental history. For instance, these methodologies can construe whether nearby
transformation results from hereditary tradeoffs (opposing pleiotropy), where local alleles are
best adjusted to neighborhood conditions, or if neighborhood variation is brought about by
contingent nonpartisanship at numerous loci, where alleles show wellness contrasts in a single
climate, however not in a differentiating climate. Biological hereditary qualities in characteristic
populaces of lasting or outcrossing plants can likewise contrast significantly from model
frameworks. In this survey of the developmental hereditary qualities of plant transformation, we
underscore the significance of field reads for understanding the developmental elements of
model and nonmodel frameworks, feature a key life history characteristic (blossoming time) and
talk about arising preservation issues.
KEYWORDS:- Hereditary, developmental, pleiotropy, variation, blossoming, preservation,

loci etc.
ISBN: 978-81-948129-1-3 Page 183

A POPULACE HEREDITARY QUALITIES (GENES) PERSPECTIVE
ON CREATURE TAMING
Shubham
Mobile no.-8708075152
ABSTRACT
In this paper, we will mainly focused on the crucial move related with the taming of plants
and creatures took into account a sensational expansion in human populace sizes and the rise of
present day society. In spite of its significance and the times of examination gave to
contemplating it, questions with respect to the starting points and cycles of taming remain. Here,
we audit ongoing hypothetical advances and present a point of view that underscores the pivotal
job that populace admixture has played in impacting the genomes of homegrown creatures in the
course of the last 10 000 years. We at that point examine novel ways to deal with producing and
dissecting hereditary information, underscoring the significance of an express theory testing
approach for the induction of the causes and resulting development and demography of
homegrown creatures. By applying cutting edge sequencing innovation close by fitting
biostatistical systems, a significantly more profound comprehension of training is not too far off.
KEYWORDS:- Hereditary, Populace, biostatical, genomes, qualities or Genes etc.
ISBN: 978-81-948129-1-3 Page 184

SNPS IN LEGAL HEREDITARY QUALITIES: A SURVEY ON
SNP COMPOSING STRATEGIES
Shubham
ABSTRACT
In this paper, we mainly focus on expanding interest in single nucleotide polymorphism (SNP)
composing in the legal field, not just for the handiness of SNPs for characterizing Y chromosome
or mtDNA haplogroups or for investigating the topographical birthplace of tests, yet additionally
for the possible uses of autosomal SNPs. The premium of legal analysts in autosomal SNPs has
been pulled in because of the possible preferences in paternity testing due to the low
transformation rates and uniquely in the investigation of corrupted examples by utilization of
short amplicons.
New SNP genotyping strategies, sciences and stages are constantly being created and it is
frequently hard to stay up with the latest and to settle on the best innovation choices accessible.
This audit offers to the peruser a best in class of SNP genotyping advances with the points of
interest and detriments of the various sciences and stages for various criminological
prerequisites.
KEYWORDS:- Single nucleotide polymorphism (SNP), chromosome, genotype, hereditary,

amplicons etc.
ISBN: 978-81-948129-1-3 Page 185

METASTASIS QUALITIES: A MOVEMENT PUZZLE
Shubham
Most, if not all, human tumors create through a progression of hereditary and epigenetic
changes that present progressively neoplastic (malignancy like) qualities on cells. To be sure,
this multistep cycle has been compared to darwinian development inside the microcosm of living
tissues, in which the units of choice are singular cells. A cell that has profitable qualities (ones
that favor endurance and expansion) is chosen to turn into the ancestor of a replacement cell
populace that at last rules the tumor mass. An uncommon variation that emerges among the
numerous replacement cells will, thusly, start the following round of clonal progression.
Somewhere in the range of six and ten such clonal progressions might be needed to create
exceptionally dangerous human malignancy cells.
As per the predominant thinking, this cycle of tumor movement gives rise at first to cells
that have procured the capacity to frame essential tumor masses of considerable size. Hereditary
changes, procured by cells during the underlying periods of tumor movement, that give some
kind of proliferative favorable position empower the phones conveying these freak alleles to
bring forth enormous relative populaces inside the essential tumor mass. Among these profitable
aggregates are the securing of constitutive mitogenic signals, the capacity to oppose development
repressing signs, to dodge modified cell passing (apoptosis) and to instigate vein development
(angiogenesis). Thusly, singular cells in these huge cell populaces procure yet more freak alleles
that empower them to metastasize to seed new provinces at anatomical locales that might be far
taken out from the essential tumor mass.
This model of tumor movement conveys with it a striking calculated irregularity: the
qualities that indicate the last advance in tumor movement — metastasis — would not appear to
give expanded proliferative advantage at the essential site. That is, there is no motivation to
believe that a metastatic aggregate empowers cells to multiply more adequately inside the
essential tumor mass, along these lines expanding their portrayal in the general tumor-cell
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populace. Henceforth, uncommon cells in the essential tumor mass that end up securing
metastatic capacity will stay uncommon. As the achievement pace of individual cells
undertaking metastasis is remarkably low, this makes it hard to envision how metastasis can
actually continue.
Thinking like this drives us to think about a very unique robotic model: specifically, that
the inclination to metastasize is generally dictated by the personalities of freak alleles that are
obtained moderately right on time during multistep tumorigenesis. It is as of now obvious that
there are a few option hereditary ways that cells can take on the way to framing an essential
tumor. Consequently, a specific aggregate required right on time in tumorigenesis by a
developing tumor cell can be obtained through the transformation of any of a few elective
development controlling qualities. We recommend that a subset of the freak alleles procured by
beginning tumor cells ahead of schedule in tumorigenesis give the chose replicative favorable
position, yet additionally, later in tumorigenesis, the proclivity to metastasize. This proclivity
will become show just a lot later in tumor movement, with regards to yet different changes that
have struck the genomes of relative cells.
This sort of reasoning has three ramifications. To begin with, the propensity of a tumor in
the long run to metastasize is now pre-appointed by the range of transformations that begetter
cells gain moderately right on time in tumorigenesis; that is, a few malignant growths begin 'off
kilter'. Second, qualities and hereditary changes explicitly and only associated with arranging the
cycle of metastasis don't exist. All things being equal, the qualities for metastasis are to a great
extent those that disease researcher have been reading seriously for an age: the oncogenes and
tumor-silencer qualities. Third, on the grounds that significant segments of the genotype of
metastasis are as of now embedded in cells generally ahead of schedule in tumorigenesis, even
moderately little essential tumor cell populaces may as of now can dispatch metastatic pioneers
to inaccessible destinations in the body.
A few free lines of proof appear to help these thoughts. In some little, all around limited
essential human bosom malignant growths, singular carcinoma cells are unmistakably
recognizable in the bone marrow. Besides, DNA-microarray investigation uncovers that the
quality articulation example of metastatic tumor cells is regularly strikingly like that of the
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phones restricted to the essential tumor mass from which they were inferred, suggesting that the
prevailing cell populace in the essential tumor mass is phenotypically and conceivably
genotypically (nearly) indistinguishable from the phones in the metastases.
Similarly important are different investigations in which the quality articulation profiles
of the prevailing populaces of bosom malignant growth cells inside an essential tumor mass have
been utilized to anticipate, with 90% precision, regardless of whether the tumor will stay limited
or whether the patient will encounter metastases and illness backslide. Here, indeed, the
metastatic conduct of these malignant growth cells is by all accounts decided generally right on
time in tumorigenesis. (The suggestions for the handiness of early clinical identification of
bosom malignancy are disrupting.) At long last, a few all around examined oncogenes, including
ras and myc, the proliferative forces of which are very much reported, can work in certain mouse
models of tumorigenesis to empower disease cells to metastasize.
These thoughts have suggestions for our possible comprehension of the hereditary and
biochemical bases of metastasis. A few specialists have looked all over in the genomes of cutting
edge, exceptionally forceful tumor cells for the qualities answerable for actuating metastatic
ability. Maybe the guilty parties have been gazing us in the face for quite a while — the freak
qualities that are known to present darwinian specific points of interest from the beginning might
be similar qualities that, sometime later, engage metastasis.
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FACILE SYNTHESIS OF FES2/AG2MOO4 NANO-HETEROSTRUCTURE FOR
PHOTOCATALYSIS AND BACTERICIDAL APPLICATIONS
S. Kokilavani, S. Sudheer Khan
Nanobiotechnology Laboratory, Department of Bio Technology,
Bannari Amman Institute of Technology, Sathyamangalam, Tamil Nadu, India.
*Corresponding Author: S. Sudheer Khan; Email: sudheerkhan@bitsathy.ac.in;
ABSTRACT
The novel, highly efficient and reusable photocatalyst was prepared successfully by chemical
co-precipitation method. The NCs were analyzed by UV-vis-DRS, XRD, BET surface area
analyzer, HRTEM, EDS and FTIR. UV–vis spectra changes of MB dye irradiated under visible
light were monitored. The degradation rate constant (k value) for the MB photocatalysis was
0.1106 min-1 by FeS2/Ag2MoO4 which was 3.45 times and 1.9 times higher than
Ag2MoO4(0.0315 min-1) and FeS2 (0.064174 min-1) respectively. The degradation efficiency of
NCs was 2 to 2.5 times boosted than its pure individual parts. Half life of dye was 76 min when
25 mg/L MB dye is degraded with 0.3 mg NCs. The photodegradation mechanism was
elucidated with a series of radical scavengers found that predominantly hydroxyl radicals as the
active species. The NCs exhibited a stable photocatalysis performance after six cycles. The
FeS2/Ag2MoO4 NCs exhibited excellent antimicrobial activity against Escherichia coli and
Bacillus subtilis. Hence, the investigation reports NCs for the effective degradation of dyes
through photocatalysis and found to act as a good antibacterial agent.
KEYWORDS: Photocatalysis; FeS2/Ag2MoO4; scavenging activity; Anti-microbial activity
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NOVEL GRAPHENE QUANTUM DOTS: A REVIEW ON SYNTHETIC TECHNIQUES

AND APPLICATIONS
Sridhar Arelli1, Akshay.S2 and Niti Chawla3
1
Department of Physics, OPJS University, Churu, Rajasthan. Mail: usri89760@gmail.com
2
Department of Physics, Maharani Lakshmi Ammanni College for Women Autonomous,
Malleswaram, Bengaluru-560012, Karnataka, Email: akshay@mlacw.edu.in
3
Department of Biotechnology, Chaudhary Bansi Lal University, Bhiwani
Email: nitichawlahans@gmail.com
ABSTRACT:
As a new class of fluorescent carbon materials, graphene quantum dots (GQDs) have
attracted tremendous attention due to their outstanding properties and potential applications in
biological, optoelectronic, and energy-related fields. Herein, top-down and bottom-up strategies
for the fabrication of GQDs, mainly containing oxidative cleavage, the hydrothermal or
solvothermal method, the ultrasonic-assisted or microwave-assisted process, electrochemical
oxidation, controllable synthesis, and carbonization from small molecules or polymers, are
discussed. Different methods are presented in order to study their characteristics and their
influence on the final properties of the GQDs. The respective advantages and disadvantages of
the methods are introduced. With regard to some important or novel methods, the mechanisms
are proposed for reference. Moreover, recent exciting progresses on the applications of GQD,
such as sensors, bio-imaging, drug carriers, and solar cells are highlighted. Finally, a brief
outlook is given, pointing out the issues still to be settled for further development. We believe
that new preparation methods and properties of GQDs will be found, and GQDs will play more
important roles in novel devices and various applications.
KEYWORDS: bottom-up; graphene quantum dots; photoluminescence; quantum confinement
effect; top-down.
1 INTRODUCTION
Graphene, discovered by Novoselov et al. in 2004 [1], is a new kind of nanomaterial
with excellent mechanical, electrical, thermal, and optical properties [2–5], following the zero-
dimensional fullerenes [6, 7] and the one-dimensional carbon nanotubes [8–11]. Many synthesis
methods have been developed, such as micromechanical stripping [1, 12], chemical vapor
deposition (CVD) [13–15], SiC epitaxial growth [16–18], and graphene oxide (GO) reduction
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[19, 20]. Because of the large plane conjugate structure of graphene, the π electrons have a
significant delocalization effect. As a result, graphene possesses a zero band gap which is the
feature of half-metallic materials, limiting its application in the fields of optoelectronic devices
and semiconductors [21]. Graphene quantum dots (GQDs), which are small pieces of graphene
with a two-dimensional lateral size (less than 100 nm), have been developed in recent years [22–
24]. Strictly speaking, ideal GQDs have a single atomic layer and only contain carbon. Actually,
most of the prepared GQDs also contain oxygen and hydrogen, and often have multiple atomic
layers, with sizes being less than 10 nm [25, 26]. The band gap energy of the GQDs can be
regulated from 0 to 6 eV by changing the two-dimensional size or surface chemical properties,
due to the quantum confinement effect of conjugated π-domains and the edge effect [27–30].
High-resolution transmission electron microscopy (HRTEM) observations show that the GQDs
have the hexagonally symmetrical crystalline structure, and the in-plane lattice spacing is 0.24
nm, completely the same with graphene [31–33]. Fourier transform infrared (FT-IR)
spectroscopy and X-ray photoelectron spectroscopy (XPS) tests demonstrate that the hydrogen
and oxygen in the GQDs mainly exist in the form of oxygencontaining groups, such as hydroxyl,
carbonyl, carboxyl, and epoxy groups [32, 34]. At present, a lot of research work is devoted to
the surface chemical modification of the GQDs in order to regulate the properties for the
applications [35–37]. The main approaches involve the control of oxidation degree [38–41],
surface functionalization [35, 42–44], and heteroatom doping [32, 45–47].
Compared to the quantum dots of a traditional semiconductor, the GQDs possess many
advantages, such as stable fluorescence properties [48–51], low toxicity [40, 52–54], and good
water solubility [55, 56]. Among them, the fluorescence properties are the most important feature
of the GQDs. Although the photoluminescence (PL) mechanism of the GQDs can be explained
by the size of the GQDs, surface chemical groups, and doping atoms, there is no universal
agreement on the specific PL mechanism. The dominant PL mechanism of GQDs consists of the
quantum confinement effect of conjugated π-domains, the surface/edge state in GQDs, as well as
the synergistic effect of these two factors [57]. Their fluorescent emission wavelength can be in
various regions, including deep ultraviolet light [49], blue light [50, 51], green light [39, 58],
yellow light [52, 59], and red light [27, 60]. It is attributed to the different sizes, surface
functional groups, and excitation wavelength.
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Besides, though the GQDs synthesized by various methods have different ultraviolet-
visible absorption characteristics, the π-π* transition absorption peak ranges commonly from
200- to 270-nm wavelength, and the n-π* transition absorption peak is more than 260 nm [55,
59]. The ultraviolet-visible absorption is dependent on the size of the GQDs, due to the quantum
confinement effect [31, 61]. In addition, oxygen-containing functional groups on the surface of
the GQDs also have an important influence on the absorption peak position [62].
Moreover, GQDs have a better potential application in biological medicine than graphene
or GO, attributed to the smaller two-dimensional nanosize [63]. Naturally, the biocompatibility
and toxicity of GQDs are the major concerns. There has been some research that indicate that
GQDs have relatively good biocompatibility and low biological toxicity [40, 52–54, 64]. In
addition, GQDs have a single atomic layer plane conjugate structure, a large specific surface
area, and oxygen-containing groups on the surface, which can provide the active sites, easily
carrying or loading drug molecules. Therefore, GQDs possess some unique properties different
from graphene and GO. To date, there have been several reviews about GQDs, due to the
significance and urgency of the research on them. For instance, Li et al. [65] introduced potential
methods like acid oxidation, hydrothermal and solvothermal reactions, etc. for the preparation of
GQDs. They emphatically discussed four representative types of biomedical application based on
GQDs, bioimaging, biosensing, drug delivery, and antimicrobial materials in detail. Li et al. [66]
summarized the synthetic methods on the top-down routes, which possess the advantages of
abundant raw materials, large-scale production, and simple operations, and systematically
discussed the optical properties of GQDs, ranging from the mechanism and the influencing
factors to the optical tunability. This work will be very useful in quickly gaining knowledge and
experience in creating novel functional GQD-based nanomaterials for further applications in
biomedicine, materials science, analytical science, and optical nanodevices.
In this article, the latest research progresses on the preparation and applications of the
GQDs are reviewed. Top-down and bottom-up strategies, which mainly contain oxidative
cleavage [34, 67], the hydrothermal or solvothermal methods [68, 69], ultrasonic-assisted or
microwave-assisted processes [70, 71], electrochemical oxidation [72, 73], controllable synthesis
[74], and carbonization [75, 76] from small molecules or polymers, are discussed. Different
methods are presented in order to study their characteristics and the influence on the final
properties of the GQDs. The respective advantages and disadvantages of the methods are
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introduced. The mechanisms of some novel methods are proposed for reference. Additionally,
the potential applications of the GQDs, such as sensors, bio-imaging, drug carriers, and solar
cells are comprehensively introduced. Finally, a brief outlook is given, pointing out the issues
waiting to be settled for further development. The current review demonstrates that the GQDs
exhibit superior performances and will be applied in many fields of new materials and novel
devices.
2 SYNTHESIS METHODS OF GQDS
In detail, the synthesis methods of the GQDs can be classified into two categories: top-
down strategy and bottom-up strategy (as shown in Figure 1) [57]. The former is extensive in the
synthesis of GQDs because it is simple and effective. GQDs are gained from carbon materials,
including graphene, fullerenes, carbon nanotubes, etc., by cutting them via chemical or physical
methods, such as oxidative cleavage, hydrothermal or solvothermal method, electrochemical
oxidation, ultrasonic-assisted or microwaveassisted process, chemical vapor deposition (CVD),
and laser ablation. For the latter, GQDs are fabricated through controllable synthesis or
carbonization from suitable organic molecules or polymers. The controllable synthesis is precise
but complicated, needing many steps of synthesis to obtain GQDs of large dimension. In the
carbonization method, GQDs are obtained from suitable small molecules or polymers by
dehydration and further carbonization. These formation processes are usually uncontrollable,
resulting in the preparation of GQDs with polydispersity.
Figure: 1 SYNTHESIS METHODS OF GQDS
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2.1 Top-down strategy 2.1.1 Oxidative cleavage Oxidative cleavage, the most widely used
method, is also named oxidation cutting. In this method, the carboncarbon bonds of graphene,
GO, or carbon nanotubes are generally broken by H2 SO4 , HNO3 , or other oxidizers.
For instance, in the work of Shen et al. [34], GO sheets with a two-dimensional size of
micron grade were put in HNO3 and cut into the smaller pieces. The products were treated with
surface passivation by ethylene glycol and then were reduced by hydrazine hydrate. Eventually,
the GQDs with a diameter distribution from 5 to 19 nm were gained. It indicates that blue
fluorescence is obtained when the light of a 365-nm wavelength is used to excite the GQDs. It is
interesting that green fluorescence can be available when the light of a 980-nm wavelength is
used. It demonstrates that the as-prepared GQDs have upconversion fluorescence properties. The
energy level structural models of GQDs are proposed to explain the process of the formation of
fluorescence and upconversion. This work presents a facile method by utilizing hydrazine
hydrate to reduce GO with surface passivation for the fabrication of GQDs with frequency
upconverted emission. The GQDs may provide a new type of fluorescence and upconversion
material for applications in bioscience and energy technology, and may expand the application of
graphene-based materials to other fields.
Via et al. [67] pressented a simple and controlled method to tune well the lateral size of
GO at the nanometer scale. The procedure is shown in Figure 2. First, GO was synthesized from
expandable graphite by the modified Hummers method. Then, GO was dispersed into deionized
water followed by sonication to obtain a brown GO solution. Later, H5 IO6 were added into the
GO dispersion, and the mixed solution was kept at 60°C for 24 h. The precipitate was
centrifuged out from solution and was then washed with deionized water until the supernatant
was neutral. After that, sodium polystyrene
Figure 2: Schematic diagram of the synthesis of GQDs for electrochemical detection
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Finally, l-ascorbic acid (l-AA) was added into the aqueous solution and stirred at 50°C
for 24 h. When the reaction ended, the color of the solution turned from yellow to dark black,
which was a visual indication that the GO nanosheets were successfully reduced into GQDs with
a mixture of different sizes (5–15 nm). The as-prepared GQDs show a strong and excitation-
independent PL activity, with the maximum emission wavelength at 470 nm. The excitation-
independent emission of the GQDs implies that both the size and the surface state of those sp2
clusters contained in GQDs are uniform. In addition, the GQDs show an enhanced performance
in the electrochemical sensing of heavy metal ions. A remarkably low detection limit of 7 × 10−9
m for Pb2+ is achieved. It is because the GQDs with a high specific surface area can improve the
interaction opportunity of active sites and target ions greatly, which endows the capability of
accumulating metal ions. The GQDs will be believed to advance the development of not only
graphene-based nanosensors but also biomaterial field.
In addition to graphene or GO, other carbon materials, such as fullerenes C60, carbon
nanotubes, and carbon fibers, can also be utilized to fabricate GQDs. Fullerene was used as a
starting material to produce very small GQDs (2–3 nm) by Pumera et al. [77], due to its well-
defined dimension. Treatment of fullerene with a mixture of strong acid and chemical oxidant
induced the oxidation, cage-opening, and fragmentation processes of fullerene. The products
remained fully dispersed in aqueous suspension and exhibited strong luminescence properties,
with the highest intensity at 460 nm under a 340-nm excitation wavelength. Further chemical
treatments with hydrazine hydrate and hydroxylamine produced GQDs and resulted in red shift
and blue shift of the luminescence, respectively. The present work has demonstrated the
simultaneous oxidation and cage opening of fullerene to provide GQDs. The simplicity of this
method in producing GQDs shows potential for further development for integration into practical
devices or applications including optoelectronics and biological labeling.
The GQDs with fluorescent properties were obtained via oxidation cutting carbon fiber
by thick H2 SO4 /HNO3 under high temperature [31]. The scheme of the procedure is shown in
Figure 3A. The as-synthesized GQDs are highly soluble in water and other polar organic
solvents, such as dimethylformamide (DMF) and dimethyl sulfoxide (DMSO). Figure 3B is the
transmission electron microscopy (TEM) image of the fiber-derived GQDs, showing relatively a
narrow size distribution between 1 and 4 nm, which are summarized in Figure 3D. The high
resolution (HR)-TEM image (inset of Figure 3B indicates
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Figure 3: Illustration of the preparation process of GQDs by oxidation cutting showing both
their morphology and size distribution. (A) Representation scheme of oxidation cutting of CF
into GQDs. (B) TEM images of GQDs (synthesized reaction temperature at 120°C); inset of (B)
is the HRTEM of GQDs. (C) AFM image of GQDs. (D) Size and height distribution of GQDs
(reprinted/reproduced with the permission of Ref. [31], copyright 2012, American Chemical
Society).
The atomic force microscopy (AFM) image in Figure 3C demonstrates the topographic
morphology of GQDs, the heights of which are between 0.4 and 2 nm, corresponding to one to
three graphene layers. The unzipping mechanism consists of the breakup of the fiber structure
and the lining up of chemical functional groups (such as epoxy or carbonyl groups) on planar
graphitic domains, causing the domains to be prone to fracture, preferably in the zigzag
direction. In addition, the research indicates that the size of the GQDs varies with the reaction
temperature, and the emission color and the bandgap of GQDs can be controlled accordingly.
The as-prepared GQDs show low cytotoxicity and excellent biocompatibility; thus, they can be
used as an eco-friendly material in biolabeling and bioimaging.
The GQDs were synthesized using acid treatment and chemical exfoliation of multi-
walled carbon nanotubes (MWCNTs) [78]. The as-prepared GQDs have a uniform size
distribution, zigzag edge structure, and two-dimensional morphology. It demonstrates that the
size of the GQDs is less than 5 nm, and the lattice parameter is estimated to be about 0.25 nm.
The results of the research indicate that the GQDs have a bright blue emission upon UV
excitation and exhibit high water solubility and good stability. It isshown that the acid treatment
of MWCNTs leads to the formation of the functional group in zigzag sites, which results in the
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pH-dependent fluorescence of the GQDs. This work presents a facile and effective method to
obtain GQDs for application in optoelectronics, electronic devices, and biological probes.
Oxidative cleavage or oxidation cutting is simple and effective in the preparation of GQDs, but
there are some defects that the strong oxidizer used may cause burning or explosion and the post-
processing process is more complicated. Therefore, some new oxidation cutting methods are
developed to settle the problems. For instance, Lu et al. [79] utilized black carbon as a precursor
and H2 O2 as a facile oxidant and developed a simple and facile one-pot method for the
synthesis of GQDs without using a strong concentrated acid, and the entire synthetic process
took only 90 min. In the preparation, black carbon was utilized as a precursor, and H2 O2 was
used as an oxidant to cut the black carbon. The only reaction products were H2 O and GQDs.
Thus, the proposed synthetic method not only avoids the use of a strong concentrated
acid and the introduction of metal impurity contamination but also does not need any other post-
processing steps. The mechanism proposed is that the free radicals (e.g. ·OH and ·O−), which are
produced from the decomposition of H2 O2 , have high reactivity and strong oxidizing properties
so they can oxidizeand cut the graphene structure of black carbon effectively. The as-synthesized
GQDs with diameters ranging from 3.0 to 4.5 nm have robust photo-stability, good resistance to
salt solution, low toxicity, and excellent biocompatibility. As a satisfactory FL probe, the GQDs
have been successfully applied in FL imaging of HeLa cells directly. Compared with other
reported methods, it is the most green and fastest synthesis method for GQDs synthesis to date.
The present study provides a green and fast method for the synthesis of GQDs and facilitates
GQDs for bioimaging and related biological applications.
2.1.2 HYDROTHERMAL/SOLVOTHERMAL METHOD
The hydrothermal or solvothermal method is a simple and rapid method for the
preparation of GQDs. It cut carbon materials into GQDs under the conditions of high
temperature and high pressure in the process. Generally, the carbon materials need to be treated
through strong oxidation before the reactions happen [42, 80, 81].
The GQDs were prepared from GO as a raw material via the hydrothermal method for the
first time by Pan et al. [68]. The procedure is as follows: first of all, GO was mixed in thick H2
SO4 /HNO3 acid for oxidation, and then, the hydrothermal reaction started under alkaline
conditions for about 10 h. Eventually, the GQDs with a size distribution of 5–13 nm were
gained. With the reaction mechanism, it was proposed that mixed epoxy chains composed of
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fewer epoxy groups and more carbonyl groups may exist in the oxidized graphene sheets (GSs),
as illustrated in Figure 4A (left). The presence of these linear defects makes the GSs fragile and
readily attacked. Some ultrafine pieces surrounded by the mixed epoxy lines and/ or edges may
further break up during the hydrothermal deoxidization process, by which the bridging O atoms
in the epoxy lines are removed, and thus, the GQDs form eventually (Figure 4A, right).
Structural models of GQDs in acidic and alkali media are also proposed based on the reversible
protonation of carbene-like zigzag sites and the reversible protonation of oxygen-containing
functional groups (Figure 4B). Because the triple carbenes are most common at zigzag edges, the
two electronic transitions of 320 nm (3.86 eV) and 257 nm (4.82 eV) observed in the PL spectra
can be regarded as transitions from the σ and π orbitals (highest occupied molecular orbitals,
HOMOs) to the lowest unoccupied molecular orbital (LUMO), as demonstrated in Figure 4C.
The work has developed a hydrothermal method for cutting preoxidized GSs into ultrafine GQDs
with strong blue emission.
Figure 4: The mechanism for the hydrothermal method and models of the GQDs in acidic and
alkali media, as well as typical electronic transitions of triple carbenes at zigzag sites. (A)
Mechanism for the hydrothermal cutting of oxidized GSs into GQDs: a mixed epoxy chain
composed of epoxy and carbonyl groups (left) is converted into a complete cut (right) under the
hydrothermal treatment. B) Models of the GQDs in acidic (right) and alkali (left) media. The two
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models can be converted reversibly depending on pH. The pairing of σ( ·) and π(o) localized
electrons at carbene-like zigzag sites, and the presence of triple bonds at the carbyne-like
armchair sites are represented. (C) Typical electronic transitions of triple carbenes at zigzag sites
observed in the optical spectra (reprinted/reproduced with the permission of Ref. [68], copyright
2010, Wiley-VCH Verlag GmbH & Co)
The discovery of the new PL from GQDs may expand the application of graphene-based
materials to other fields such as optoelectronics and biological labeling. Figure 5 presents the
hydrothermal strategy for the production of amino-functionalized GQDs (af-GQDs) by Tetsuka
et al. [42]. The procedure is as follows: first, homogeneous oxidized graphene sheets (OG) (OG
was synthesized according to the modified Hummers method, and all of other agents were
obtained from Wako Pure Chemical Industries) water dispersion was mixed with ammonia
solution and was heated at 70–150°C for 5 h in an autoclave. After cooling to room temperature,
the precipitated insoluble large fragments were filtered out using a 0.02-μm Anopore inorganic
membrane (AnodiscTM, Watman). The desired af-GQDs were present in the supernatant. Then,
the yellow supernatant was heated and ultrafiltered through centrifugal filter device (Amicon
Ultra-4, Millipore).
Figure 5: The hydrothermal strategy for the production of amino-functionalized GQDs (af-
GQDs) by Tetsuka et al. [42]. (A) Schematic illustration of the preparative strategy for af-GQDs.
(B) C1s and N1s X-ray photoelectron spectra for af-GQDs prepared at 90°C (green line) and
OGS (gray line). (C) Transmission infrared-absorbance spectra of af-GQDs prepared at 90°C
(green line) and OGS (gray line) (reprinted/reproduced with the permission of Ref. [42],
copyright 2012, Wiley-VCH Verlag GmbH & Co).
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The obtained yellow suspension was further dialyzed through a dialysis tubing membrane
to remove the remaining tiny fragments. As shown in Figure 5A, after being oxidized, the GO
starts the hydrothermal reaction in the ammonia system. It is found that the GO is cut into GQDs,
and amino substitution reaction happens at the same time. The GQDs with the amino group are
gained at last, as proven in Figure 5B and C. The production of uniform af-GQDs with a
diameter of ~2.5 nm is verified by TEM. Aqueous suspensions of af-GQDs exhibit bright
colorful luminescence under irradiation from a UV lamp with a wavelength of 365 nm. Their
corresponding emission peaks vary with the quantity of amine functionalization in the range of
420–535 nm and retain a sharp full width at half maximum (FWHM) as narrow as ~65–80 nm.
The research demonstrates a novel strategy for the preparation of GQDs with high efficiency and
wide tunability of narrow photoluminescence lines under a single-wavelength UV excitation,
whose energy is controlled by their edge-termination structure with primary amines. Their
superior optical properties should enable the use of af-GQDs in numerous applications, including
multicolor light-emitting devices, biological applications, and photovoltaics.
Significantly, Tian et al. [69] reported a one-step solvothermal method for synthesizing
GQDs with the application of hydrogen peroxide in a DMF environment, which introduces no
impurity in the whole preparation process. The process is shown clearly in Figure 6. Typically, a
muffle furnace was first heated to 800°C for 5 h, and then, 2 g of expandable graphite in an
alumina crucible was placed in a high-temperature environment for 10 s to form an expanded
graphite. In this step, sulfuric acid and nitric acid molecules decompose into gas and escape from
the interlayer of the expanded graphite. The expanded graphite was then mixed with DMF and
treated with ultrasound to remove air in the layered structure of the expanded graphite, making
full contact between the solvent and the expanded graphite. Subsequently, hydrogen peroxide
was added into the mixture and stirred for 5 min to form a homogeneous solution.
The final mixture was transferred into an autoclave and heated up to 170°C for 5 h. The
product solution was treated with vacuum filtration to obtain GQDs. The mechanism of the
reaction is similar to electrochemistry exfoliation of graphite anode: water is oxidized under high
redox potential to generate oxygen and hydroxyl radicals, which act as the “scissors” to cut down
the graphite anode. It indicates that the diameters of the GQDs were mainly distributed in a range
of 20–40 nm, and the thickness mainly falls in the range of 1–1.5 nm, corresponding to two to
three graphene layers. As the excitation wavelength increases from 280 nm to 420 nm, the PL
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intensity increases to maximum and then decreases, while the PL peaks shift from 398 nm to
480 nm. The quantum yields of 15% in neutral conditions and good photoluminescence stability
in different pH conditions are demonstrated, implying a wider application in different harsh
environments.
This is the first time that GQDs were prepared from graphite materials by hydrogen
peroxide without a dialysis process. Because of the extremely low cost, good water solubility,
high quantum yield, no need for dialysis for purification, and easily-obtained experiment
equipment, this method shows great promise in the biomedical field and electronic device.
It is noteworthy that Liu et al. [82] reported a facile one-pot synthesis method using graphite as
the starting material, which can selectively obtain either pure GO or pure GQDs within 2 h, with
a higher production yield (93% for GO and 10% for GQDs). The synthesis process is elaborated
in Figure 7, that is, graphite powder is mixed with potassium permanganate, and sulfuric acid is
gradually added under magnetic stirring. Then, nitric acid is introduced into the autoclave, which
is heated to a certain temperature for the desired products.
It proves that the size (from 50 to 2.5 nm) of the GQDs can betuned by simply varying
the ratio of graphite and potassium permanganate. The emission spectra of the 2.5-nm GOQDs
demonstrate a perfect excitation-dependent photoluminescence behavior. The strong
luminescence emission suggests a high luminescence yield (about 8.8%) and, sequentially, a
high quality of the as-synthesized GOQDs. It should be mentioned that typical solvothermal
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methods, utilizing graphene oxide as a precursor, need more complicated processing when
compared with this strategy. This one-pot synthesis method has the following unique
characteristics: (1) the operation procedure is simplified because the reaction happens in a sealed
autoclave; (2) GO or GQDs can be selectively obtained by simply adjusting the reaction
temperatures; (3) the size of the GQDs can be easily tuned from 50 to 25, 13, 5, and finally to
2.5 nm, with strong luminescence emission; (4) various carbon sources, including MWCNTs,
single walled carbon nanotubes (SWCNTs), carbon black, etc.,
2.1.3 MICROWAVE-ASSISTED/ULTRASONIC-ASSISTED PROCESS

As we know, one of the common problems is the long reaction time when the oxidative
cleavage or hydrothermal/ solvothermal method is used. The microwave technique, as a rapid
heating method, is widely used for the preparation of nanomaterials. It not only can shorten the
reaction time but also improve the production yield. Accordingly, electro chemiluminescent
(ECL) two-color GQDs were fabricated by the cleavage of GO in the acidic condition via the
microwave-assisted method by Li et al. [38]. The oxidation reaction was shortened, and the
yield was improved to around 8%. Figure 8 outlines the preparation route to greenish-yellow
luminescent GQDs (gGQDs, unreduced) and bright blue luminescent GQDs (bGQDs, further
reduced with NaBH4 ).
For the preparation of stabilizer-free gGQDs, GO nanosheets were kept under acid
conditions (3.2 m HNO3 and 0.9 m H2 SO4 ) within 3 h. Then, bGQDs were obtained via
moderately reducing gGQDs with NaBH4 within 2 h. It indicates that the diameters of gGQDs
are mainly distributed in the range of 2–7 nm withan average diameter of 4.5 nm, and the
dimensions and height of the bGQDs show no perceptible change, demonstrating that the PL
blue shift of the bGQDs is attributed to their structural change after reduction rather than their
dimension variation. The PL quantum yields of gGQDs and bGQDs are as high as 11.7% and
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22.9%, respectively. Briefly, the cleaving mechanism is described as follows: initiated by the
acid oxidation of epoxy groups, it is prone to form a mixed line on the carbon lattice composed
of fewer epoxy groups and more carbonyl groups, making the graphitic domains fragile and
readily attacked. These kind of ECL-active GQDs are expected to have promising applications in
the development of novel ECL biosensors due to their low cytotoxicity, low cost, excellent
solubility, and ease of labeling.
The GQDs were synthesized by the microwaveassisted pyrolysis method using the
following procedure by Zhang et al. [70]. In the preparation, aspartic acid (Asp) and NH4 HCO3
were added to 20 ml of deionized water in a beaker and then were heated using microwave
irradiation for 10 min. The products were further purified by dialyzing against deionized water
using a membrane with a dialysis membrane for 7 h to obtain the final GQDs. The diameters of
the GQDs are mainly distributed in the narrow range of 1.8–2.4 nm with an average size of
2.1 nm. The solution of the GQDs emits bright blue luminescence under the irradiation of UV
light (365 nm). The as-synthesized GQDs possess lower cellular toxicity and high photostability,
and is very sensitive not only to the pH value but also to iron ions. Therefore, the GQDs can
serve as a fluorescent probe for the sensitive detection of Fe3+ and pH value and can be directly
used for live cell imaging.
Besides, the ultrasonic technique is also a commonly used method in the synthesis of
materials. Tens of thousands of small bubbles in the liquid will form in the presence of ultrasonic
waves and produce instantaneous high pressure and high energy when growing, shrinking, and
collapsing, destroying the carbon-carbon bonds. For instance, an environmental friendly, fast,
and industrial promising method for synthesizing GQDs at a large scale was reported via an
ultrasonic-assisted liquidphase exfoliation technique [71].
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The GQDs with different sizes, structures, and defect contents were obtained using
different graphitic carbon precursors for exfoliation, and the production yield of GQDs can reach
3.8 mg/ ml. In a typical preparation of GQDs, acetylene black was dispersed in N-methyl-2-
pyrrolidone (NMP) solvent. The dispersion was then under mild ultrasonication for 1 h. A gray
liquid containing dispersed GQDs and some residual precipitates was obtained. The precipitates
were removed by centrifugation at 10,000 rpm for 30 min, obtaining a homogeneous gray
dispersion of GQDs. It shows that the size distribution of GQDs is between 2 and 6 nm, and the
thickness distribution is of 0.4–2 nm, indicating a monolayer to a few layers of GQDs. The
GQDs exhibit strong excitation-dependent PL behavior. When excited at the wavelength range
from 310 to 490 nm, the PL shows red shifts from 428 to 528 nm. The present work offers a new
strategy for large-scale synthesis of GQDs with different contents of defects and edge structures,
which can be used as the initials for further functionalization in the application of biology,
electronic, energy, and engineering.
For the first time, an integrated O3 /H2 O2 /ultrasound process was proposed for the
innovative synthesis of GQDs by Wen et al. [83]. Typically, a GO aqueous suspension was
mixed with H2 O2 in a quartz tube, continuously purged with O3 for 3 h under ultrasonic
condition. Then, the nitrogen was bubbled into the resulting suspension for 15 min to terminate
the reaction.
The solid GQDs can be obtained via vacuum freeze drying. The resultant GQDs exhibit
excellent PL performances with uniform lateral dimensions of 4–10 nm. It indicates that the PL
intensities become stronger as more GO fragments are further cut into undersized pieces with the
prolonging of O3 /H2 O2 /ultrasound treating time. The possible synthesis mechanism is
proposed in Figure 9. The reaction process mainly consistsof two steps. In the first stage, the
C=C/C-C bands near the defects and sp3 carbons in GO are more prone to be oxidized and cut
down by the generated ·OH assisted by ultrasound.
The ultrasound is supposed to increase the amount of ·OH and also strengthen the
molecular vibration, thus, speeding up the GO breakage into undersized fragments. Meanwhile,
the oxidation of ·OH leads to an obvious increase in the total oxygen-containing groups upon the
GO surface. With the prolonging of the reaction time, the generated ·OH radicals accumulate and
cause further cleavage and oxidization of the GO fragments under the promotion of ultrasound,
leading to the generation of the GQDs. This study put forward a facile and efficient synthesis
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route of high-purity -COOH-enriched GQDs, and it is based on the synergetic oxidative tailoring
of GO into quantum dots by the O3 /H2 O2 /ultrasound process. Moreover, it is expected to
advance the fundamental research of quantum dots and the applications of the as-generated
GQDs in other potential fields such as solar cell, fluorescent detection, cellular imaging, and so
on.
Interestingly, a novel method that combined the merits of microwave-assisted and
ultrasonic-assisted processes was reported by Luo [84]. In the work, whitelight-emitting GQDs
(WGQDs) were prepared by a facile two-step microwave-assisted hydrothermal method (as
shown in Figure 10). The procedure is as follows: yellow-green fluorescent GQDs are
synthesized beforehand through exfoliation of oxidized graphite under the ultrasonication and
microwave irradiation. The GQDs show their average lateral size of 2.5 nm, and the average
height of the GQDs is calculated to be about 2 nm from the AFM height image. After further
reaction under microwave irradiation in alkaline solution (pH 13.0), the GQDs were converted to
WGQDs with photochemical stability and nontoxicity. As shown from the TEM images, the
lateral size of the WGQDs is still kept at about 2–5 nm. The AFM height image of the WGQDs
evaluates that the height of the WGQDs is about 1.25–2.75 nm, indicating that the as-prepared
WGQDs consisted of multilayer (two to five layer) graphene. The crystal lattice spacing of
0.335 nm can be clearly observed in the HRTEM image. When the colloidal solution of the
WGQDs is excited at 365 nm, the PL spectrum exhibits a broad emission band at 445 nm along
with a relatively weak peak at 575 nm. It can be observed that the fluorescent intensity of the
WGQDs decreases with the change in the excitation wavelength from 325 to 400 nm. The
WGQDs are subsequently used as a phosphor to fabricate a white-light-emitting diode (WLED)
device by a solution-processing method.
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Figure 9: Schematic illustration of the possible two-step generation mechanism of GQDs

(reprinted/reproduced with the permission of Ref. [83], copyright 2016, Elsevier B.V.).
Figure 10: Schematic presentation of preparing WGQDs. (reprinted/reproduced with the

permission of Ref. [84], copyright 2016, Wiley-VCH Verlag GmbH & Co).
This study presents a novel method and demonstrates that WGQDs used as a single-phase
whitelight-emitting phosphor are expected to be a highly efficient avenue to improve the
performance of WLED based on carbon nanomaterials.
2.1.4 ELECTROCHEMICAL OXIDATION

In the electrochemical oxidation method, graphite, graphene, or carbon nanotubes work
as a working electrode and are oxidatively cleaved into the GQDs under high REDOX voltage
(±1.5 ~ ± 3 V). There are two approaches for the electrochemical oxidation. One is that the
carbon-carbon bonds of graphene or carbon nanotubes are directly fractured by electrochemical
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oxidation. The other is that water is oxidized to turn into a hydroxyl free radical (·OH) or an
oxygen free radical (O·), which can oxidatively cleave them into GQDs. The GQD solutions
gained by the electrochemical oxidation method show high levels of stability, but their
disadvantage is that both the pretreatment of raw materials and the purification of GQDs
products take a long time. Moreover, it is difficult to realize the mass production of the GQDs
because of the low product yield.
In the study of Li et al. [72], the graphene filter film was used as a working electrode, and
the phosphate buffer solution (PBS) was utilized as the electrolyte to fabricate GQDs. The
electrochemical preparation of functional GQDs was performed by the CV scan within ±3.0 V at
a scan rate of 0.5 V s−1 in 0.1 m PBS. The graphene film (5 mm × 10 mm) was used as a
working electrode. Pt wire and Ag/AgCl were used as counter and reference electrodes,
respectively. Graphene film was prepared by filtration and treated with O2 plasma for seconds
prior to the preparation of the GQDs to enhance its hydrophilicity. The water-soluble GQDs
were collected after filtering and dialyzed with a cellulose ester membrane bag.
As prepared GQDs have a uniform size distribution (3–5 nm), present a green
luminescence, and can be retained stablyin water for several months without any changes. Next,
the research group replaced PBS with acetonitrile containing tetrabutylammonium perchlorate,
as the electrolyte, and made the nitrogen-doped GQDs (N-GQDs) with a size of 2–5 nm [45].
Unlike their N-free counterparts, the newly produced N-GQDs with an N/C atomic ratio of ca.
4.3% emit blue luminescence and possess an electro catalytic activity comparable to that of a
commercially available Pt/C catalyst for the oxygen reduction reaction (ORR) in an alkaline
medium. In addition to their use as metal free ORR catalysts in fuel cells, the superior
luminescence characteristic of N-GQDs allows them to be used for biomedical imaging and
other optoelectronic applications.
Similarly, Shinde et al. [73] utilized propylene carbonate containing LiClO4 as the
electrolyte and prepared GQDs by cutting MWCNTs via a two-step electrochemical oxidation.
The sizes of the GQDs can be regulated by changing the temperature and electrolytic time.
Figure 11 represents the two-step process for the electrochemical transformation of the
MWCNTs to GQDs. The first step comprises the application of a typical anodic potential of 1 V
vs. Pt QRE (quasi reference electrode) to the MWCNT-coated working electrode in propylene
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carbonate with LiClO4 , in which the applied electric field initiates the breaking of sp2 carbon
atoms. More interesting is the variation with respect to the time of oxidation and also changes in
the second step after applying a potential of −1 V vs. QRE.
This is attributed to the intercalation of Li+ /propylene carbonate complexes resulting in

exfoliation of oxidized MWCNTs, facilitating the formation of size-tunable GQDs. This work
has presented a new electrochemical approach to size-tunable GQDs [3( ± 0.3), 5( ± 0.3),
8.2( ± 0.3) nm at 90°C, 23( ± 2) nm at room temperature] from MWCNTs in a non-aqueous
solution without using any molecular capping agent for the first time. These GQDs are especially
useful for cellular and molecular imaging applications due to intrinsic luminescence behavior,
higher photostability, and enhanced fluorescence quantum yield.
2.1.5 OTHER METHODS
Because of the unique structure and excellent properties of GQDs, more and more
preparation methods have been reported, such as the chemical vapor deposition (CVD) [85–87],
pulsed laser ablation (PLA) [88–91], and electron-beam irradiation [92]. For instance, Lee et al.
[93] reported the size-controlled fabrication of uniform GQDs using self-assembled polystyrene-
b-polydimethylsiloxane (PS-PDMS) block copolymers (BCPs) as an etch mask on graphene
films grown by chemical vapor deposition (CVD). Figure 12 illustrates the chemical structure of
the BCP and a schematic of the overall process to obtain GQDs on an SiO2 /Si substrate. The
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first step of the fabrication process is the self-assembly of a PS-PDMS BCP film on a
graphene/Si/SiO2 substrate. The single-layer graphene on the Si/SiO2 substrate was prepared
using CVD and then treated with a thin (3–4 nm) hydroxyterminated PDMS homopolymer
brush. The PS-PDMS BCP was dissolved in toluene and spin-coated onto the substrate. In the
second step, oxygen plasma etching processes produced well-ordered silica dot arrays with
hexagonal symmetry, which were used as a masking layer for the patterning of the GQDs
(Figure 12B). Although adjacent silica dots were very closely packed, GQDs were overetched
depending on the condition of the oxygen plasma treatment. Electron microscope images show
that the as-prepared GQDs are composed of mono- or bilayer graphene with diameters of 10 nm
and 20 nm, corresponding to the size of BCP nanospheres. In the measured PL spectra, the
emission peak of the GQDs on the SiO2 substrate is shown to be at ~395 nm. This research
provides a noteworthy contribution to enhancing the properties of GQDs and to the
understanding of the effects of size and functionalization on GQDs.
Significantly, a facile and remarkably rapid method for production of GQDs exhibiting
excellent optoelectronic properties was reported by Kang et al. [88]. In this work, the pulsed
laser ablation technique (PLA) is employed to exfoliate GQDs from MWCNTs. The procedure is
as follows: MWCNTs were dispersed in n-hexane and Silica dot Silica dot GQD GQD Silica dot
removal Bottom PDMS PS matrix PDMS sphere Self-assembly ethanol mediums, respectively.
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Figure 12: The chemical structure of the BCP and a schematic of the overall process to obtain
GQDs on an SiO2 /Si substrate. (A) Chemical structure of the BCP. (B) Schematic illustration of
the fabrication of GQDs including the spin coating of BCP, formation of silica dots, and etching
process by O2 plasma (reprinted/reproduced with the permission of Ref. [93], copyright 2012,
American Chemical Society).
Ultrasonication was subsequently performed on the solutions for 2 h in order to achieve
homogeneous dispersion of MWCNTs. Fifty milliliters of the solution were transferred into glass
vials, and then the PLE process was performed on the fixed vials for 6 min using a Quanta Ray-
LAB 190 equipment (SpectraPhysics, USA). The laser was focused on the center of the vials
with a focal length of 10 cm and a spot diameter of 10 mm2 to prepare the GQDs. Strikingly, it
takes only 6 min to transform all the MWCNT precursors to GQDs. Furthermore, it can control
the oxidation degree of the GQDs by simply changing the organic solvents utilized in the
processing. The as-prepared GQDs show a distinct blue PL with excellent quantum yield (QY)
up to 12% as well as sufficient brightness and resolution suitable for optoelectronic applications.
This work will further open up new routes for the preparation of different optoelectronic
nanomaterials.
Moreover, in the study of Wang et al. [92], a room temperature strategy for the synthesis
of single-crystalline fluorescent GQDs via electron beam irradiation was reported. The precursor
containing high-activity nitro groups is easy to fusion GQDs with the method. Under optimized
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conditions, the GQDs exhibit highly efficient fluorescence at 475 nm with a quantum yield of
32%. The PL maximum corresponds well with the excitation wavelength. In addition, the single-
exponential fluorescence lifetime (4.86 ns) exhibits the intrinsic PL characteristic. The pH
stability in neutral and alkaline solution, solid solubility, and storage time stability of GQDs are
satisfactory. It is demonstrated that the GQDs can be applied as a safety fluorescent probe for
cell imaging.
Besides, GQDs were fabricated through grinding of graphite in ionic liquid by
Papakonstantinou et al. [94]. The size distribution of the GQDs was wide (from 2 to 29 nm), and
the thickness was of two to five atomic layers. The advantage of this method is that there is no
introduction of oxygen-containing functional groups on the GQDs, but the thickness of the
GQDs is difficult to control, and normally, multilayer GQDs are gained. Kim et al. used oxygen
plasma to cut graphene into the GQD array with templates of block copolymer. The size of the
GQDs and the array pitch can be adjusted by template morphology [95]. Lin et al. inserted
potassium atoms into the layers of MWCNTs, forming an intercalation compound and then
cleaved the MWCNTs into GQDs under the energy that was generated by violent reaction of
potassium atoms with water [55]. Kotchey et al. [96] fabricated GQDs from GO with enzyme
catalytic oxidation, with the reaction time lasting for dozens of days. GQDs were made through
the extension of oxidation time(4–5 days) via the Hummers method by Zhang et al. [97]. Small
amounts of GQDs were gained using a ruthenium catalyst for the catalytic cracking of fullerene
by Lu et al. [98]. GQDs were gained through separating different sizes of GO via density
gradient centrifugation by Sun et al. [99]. The GQDs were isolated from GO via selective
precipitation by Shi et al. [100]. However, these methods have defects of low yield, long reaction
time, or complicated process and are not popularly used.
2.2 BOTTOM-UP STRATEGY
2.2.1 CONTROLLABLE SYNTHESIS
In this method, GQDs are synthesized from phenyl-containing compounds through
stepwise controllable synthesis in the organic solvent. The as-synthesized GQDs have an
accurate number of carbon atoms, uniform size, and shape. However, the preparation process
includes multistep complicated chemical reactions, which are not only consumed for a long time
but are also of low yield. For instance, GQDs containing 168, 132, and 170 carbon atoms
(denoted by 1, 2, 3, respectively, as shown in Figure 13A) were fabricated via this method by Li
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et al. [74]. The 2′,4′,6′-trialkyl phenyl molecules (TPM) were connected to the edge of the GQDs
in order to prevent the GQDs from reuniting. The possibility of the GQDs overlapping with each
other was reduced, and the dispersibility in the organic phase was improved, because of the steric
hindrance caused by TPM stretching in three-dimensional direction. The synthesis of 1–3 is
outlined in Figure 13B. It starts from small-molecule precursors, such as 3-iodo-4-bromoaniline
(denoted by 4) and other substituted benzene derivatives, to synthesize two key intermediates
(denoted by 5 and 6, respectively). Subsequently, stepwise Suzuki coupling reactions lead to
polyphenylene dendritic precursors for 1–3, which are then exposed to an excess of FeCl3 in a
dichloromethane/nitromethane mixture, yielding the graphene quantum dots. The quantum dots 1
and 3 presented here, in particular, have large extinction coefficients in a wide spectral range
from UV to near-infrared and, thus, can serve as a new type of light-harvesting media for
photovoltaics. This work demonstrates the versatile synthesis of large, stable colloidal graphene
quantum dots with desired sizes and structures enabled by a new solubilization strategy.
Sulfur-doped GQDs (S-GQDs) with bright blue emission have been prepared from 3-
mercaptopropionic acid (MPA) and 1,3,6-trinitropyrene (TNP) by a facile one-pot method [101].
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Briefly, MPA and TNP were first mixed by ultrasonic and placed into an autoclave for
hydrothermal treatment at 200°C for 10 h. Then, the solution containing water-soluble GQDs
was filtered through microporous membrane to remove the insoluble carbon product, and un-
reacted small molecules were removed by dialyzing. After dialysis, a pale-yellow aqueous
suspension of the S-GQDs was obtained and freeze dried. The possible structure of S-GQDs is
given in Figure 14. The -SH groups of MPA add to the pyrene ring in the hydrothermal reaction.
The carboxyl groups in the MPA link to the GQDs at the same time to exhibit the -COOH
modification. Also, some S atoms may be captured and fused in the graphene lattice. The
synergistic effect of S-doping and -COOH in the GQDs may possess special coordination
abilities and corresponding selectivity toward metal ions. The S-GQDs are successfully explored
as a sensing probe for Ag+ detection with high sensitivity and selectivity. The ease of the one-
step synthesis and the promising performance of the S-GQDs make this work attractive in
bioimaging, sensing, optoelectronic devices, and catalysis.
2.2.2 CARBONIZATION
The molecular carbonization is an environmentally friendly and facile method, which
utilizes suitable organic molecules or polymers for dehydration and further carbonization [49, 57,
75, 102]. The GQDs with polydispersity are obtained because the size and the structure are
difficult to be controlled precisely for this method. Forinstance, low-cost and high-yield green-
photoluminescent single-layer GQDs were synthesized with only deionized water and glucose as
a precursor via this method [102]. In a typical synthesis, glucose powder was dissolved in
deionized water. The mixture was treated in an autoclave at 200°C for 8 h. The initial liquid
sample was transparent (colorless), and it changed to orange after the synthesis of SLGQDs. The
mechanism is presented as follows: under the hydrothermal process, glucose molecules are
dehydrated to form C=C, which is the elementary unit of the graphene structure. During the
formation of the QDs, the hydrogen atoms of a glucose molecule interact with the hydroxyl
groups of an adjacent glucose molecule leading to the formation of water molecules.
Consequently, carbon atoms covalently interact with each other, and finally, GQDs are
formed, as schematically shown in Figure 15. The SLGQDs have a uniform dispersion without
any apparent aggregation with an average size of about 8 nm. When the SLGQD solution is
excited at wavelengths from 450 to 520 nm with an interval of 10 nm, the emission peaks of the
sample at various excitation wavelengths do not shift, and the maximum emission wavelength
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remains at around 540 nm. It implies that both the size and the surface state remain uniform, and
the product is green photoluminescent. This facile method is presented to prepare the SLGQDs
from only glucose powder as a precursor in DI water, and shows the prominent advantages of
low-cost, high-yield, and large-scale production over the reported ones.
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Figure 16: Diagram for the synthesis of GQDs and GO. The black dots in the GO represent
oxygen atoms (reprinted/reproduced with the permission of Ref. [75], copyright 2012, Elsevier
B.V.)
A simple carbonization method for the preparation of both GQDs and GO has been
developed by tuning the carbonization degree of citric acid (CA) and dispersing the carbonized
products into alkaline solutions [75]. The preparation schematic of the GQDs and GO is shown
in Figure 16. In a typical procedure of the GQD preparation, 2 g of CA was put into a 5-ml
beaker and heated to 200°C using a heating mantle. About 5 min later, the CA was liquated.
Subsequently, the color of the liquid was changed from colorless to pale yellow, and then orange
in 30 min, implying the formation of the GQDs. If the heating was kept on, the orange liquid
would finally turn to black solid in about 2 h, suggesting the formation of GO. The as-prepared
GQDs are ~15 nm in width and 0.5–2.0 nm in thickness. They show a relatively strong (9.0%)
PL quantum yield and an excitation-independent PL emission activity. In contrast, the GO
nanostructures consist of sheets that are hundreds of nanometers in width and ~1 nm in height,
exhibiting a relatively weak (2.2%) PL quantum yield.
This work presents a simple bottom-up method to selectively prepare GQDs and GO by
tuning the carbonization degree of a common organic precursor CA. Additionally, Naik et al.
[103] have reported the pyrolysis of CA and further addition of sodium hydroxide for
maintaining the PH for fabrication of GQDs. The CA decomposes and thehydronium ion formed
from the acid acts as a catalyst in the subsequent decomposition reaction. Aromatization and
formation of aromatic clusters take place via aldol condensation and cycloaddition, after further
addition of sodium hydroxide. Eventually, GQDs are produced at a different pH. In this work,
the pH plays an important role in the formation of GQDs from citric acid.
Besides, there are some reports that biomass or polymers are also utilized to fabricate
GQDs by this method. In the work of Teymourinia et al. [104], the GQDs were prepared from
corn powder as a green precursor. In typical synthesis, corn powder was dispersed into DI water
and stirred, and then the solution was transferred into a Teflon-lined stainless autoclave and
heated at 180°C for 8 h. At last, the solution was centrifuged at 14,000 for 15 min to separate the
GQDs, which were uniform in size ranging from 20 to 30 nm in diameter. A broad emission
centered at 450 nm is observed in the emission spectrum (PL) with an excitation wavelength at
360 nm and the excitation spectrum (PLE) revealing a broad peak centered at 365 nm.
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This work presents novel and green methods to synthesize GQDs using corn powder.
Rice husk biomass was also utilized as an abundant source to controllably prepare high-quality
GQDs with a yield of 15 wt% [105]. The as fabricated GQDs can be stably dispersed in water,
exhibiting bright and tunable photoluminescence. It is worth noting that mesoporous silica
nanoparticles are also synthesized as a byproduct during the fabrication of GQDs. This strategy
achieves a comprehensive utilization of rice husks, exhibiting tremendous benefits on both
economy and environment. Zhou et al. [76] presented a simple and effective approach toward PL
GQDs using commercially available polycyclic aromatic hydrocarbon (PAHs) as the precursors.
The obtained PL GQDs have sizes of 5–10 nm and thicknesses of 0.5–2 nm, exhibiting excellent
water solubility, low toxicity, high optical stability, and tunable fluorescence. It was shown that
the PL GQDs could be used as fluorescent probes for highly sensitive Fe3+ and H2 O2 detection.
The present work paves the way for scalable and low-cost synthesis of PL GQDs, which will
enable further exploration of their applications in the biomedical and optoelectronic fields.
In general, the top-down strategy is extensively used in the preparation of GQDs. The
GQDs are gained from carbon materials, including graphene, fullerenes, carbon nanotubes, etc.,
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by cutting them via chemical or physical methods, such as oxidative cleavage, hydrothermal or
solvothermal method, electrochemical oxidation, ultrasonic-assisted or microwave-assisted
process, and laser ablation. For the bottom-up strategy, the GQDs are fabricated through
controllable synthesis or carbonization from suitable organic molecule or polymers. The
respective advantages and disadvantages of the different methods are summarized in Table 1.
3 POTENTIAL APPLICATIONS
3.1 SENSORS
The interaction between GQDs and certain substances can cause fluorescent intensity of
the GQDs to be reduced. According to this principle, a variety of chemical or biological sensors
can be designed to detect heavy metal ions [106, 107], small organic or inorganic molecules
[108, 109], biological molecules [110, 111], etc. For instance, Wang et al. [112] made use of
GQDs to detect Fe3+ ions. It is found that Fe3+ ions have a significant impact on the
fluorescence intensity of GQDs because of the special complexation between Fe3+ ions and the
phenolic hydroxyl groups of the GQDs. When the concentration of Fe3+ ions in the solution
arrive to 80 ppm, the fluorescence of GQDs is quenched. However, other metal ions with the
same concentration have little impact on the fluorescence intensity. It shows that the GQD-based
sensors to Fe3+ ions are highly selective. Fan et al. [108] investigated using GQDs to detect
trinitrotoluene (TNT) in solution, based on the fluorescent quenching of GQDs by TNT. It
demonstrates that TNT is adsorbed on the surface of the GQDs by a π-π interaction, and
fluorescence resonance energy transferis caused by molecular dipole-dipole interaction within a
certain distance, reducing the fluorescence intensity of GQDs. Li et al. [38] designed a new type
of electrochemical fluorescent sensors to test Cd2+ ions, taking advantage of the electrochemical
fluorescence emission properties of the GQDs.
Moreover, GQDs have excellent electrical conduction ability, good dispersibility, and a
large specific surface area, and is advantageous for the biological molecules to load on its
surface. Therefore, Shehabab et al. [110] utilized a simple route to prepare GQDs via glucose
carbonization for a nonenzymatic glucose sensor. GQDs functionalized with phenylboronic acid
(PBA) receptors were employed as a sensing material. The developed sensor has a linear
response to glucose over a concentration range of 4–40 mm with a correlation coefficient of 0.97
and a low detection limit of approximately 3.0 mm. The fluorescence enhancement after the
addition of PBA is caused by the coupled effect between the conjugated aromatic rings of the
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GQDs and PBA functional groups, as indicated in the schematic diagram of Figure 17.
Maleimide-functionalized GQDs (M-GQDs) were synthesized and deployed for biothiol
(cysteine, homocysteine, or glutathione) recognition following the principle of Michael addition
[111]. The M-GQDs probe is found to be highly sensitive and selective toward biothiol detection
in the nanomolar range inaqueous solution and at a physiological pH (7.0). Moreover, a
fluorescence immunoassay sensor was fabricated to detect immunoglobulin G (Ig G), based on
the fluorescence resonance energy transfer mechanism [113]. In the work of Liu et al. [43],
highly photoluminescent glycine (GLY)-functionalized GQDs (GLY-GQDs) were synthesized
by a simple and green pyrolysis method employing ethylene glycol as the carbon source. The as-
synthesized GLY-GQDs exhibit excellent water solubility with a fluorescence quantum yield of
21.7%. GLY-GQDs can be used as the fluorescence probe of ascorbic acid (AA), based on the
quenching effect of Ce(IV) to GLY-GQDs and the special redox reaction between AA and
Ce(IV), as shown in Figure 18.
Besides, Zhang et al. [114] fabricated a nanofibrous membrane of GQDs by
electrospinning water-soluble GQDs with polyvinyl alcohol (PVA) directly. For the first time,
the created PVA/GQD nanofibrous membrane was utilized to fabricate dual-purpose fluorescent
and electrochemical biosensors for the highly sensitive determination of hydrogen peroxide (H2
O2 )and glucose. The experimental results indicated that the fluorescence intensity of the
nanofibrous membrane decreased linearly with increasing H2 O2 concentration because the
addition of H2 O2 leads to fluorescence quenching of the GQDs.
ISBN: 978-81-948129-1-3 Page 218

Figure 18: “Off-on” fluorescence system of GLY-GQDs-Ce4+-AA (reprinted/reproduced with

the permission of Ref. [43], copyright 2017, Elsevier B.V.).
After binding glucose oxidase onto the created nanofibrous membrane, the fabricated
nanofibrous membrane showed high sensitivity and selectivity for glucose detection. In addition,
the PVA/GQD nanofibrous membrane can also be directly electrospun onto an electrode for the
electrochemical detection of H2 O2 . This novel nanofibrous membrane exhibits excellent
catalytic performance and fluorescence activity and, therefore, has potential applications for the
highly stable, sensitive, and selective detection of H2 O2 and glucose.
In the work of Lin et al. [115], photoluminescent GQDs (2–5 nm) have been used for the
first time as moleculelike building blocks to construct self-assembled hybrid materials for label-
free biosensors. Ionic self-assembly of disc-shaped GQDs and charged biopolymers is found to
generate a series of hierarchical structures that exhibit aggregation-induced fluorescence
quenching of the GQDs and change the protein/polypeptide secondary structure. The integration
of GQDs and biopolymers via self-assembly offers a flexible toolkit for the design of label-free
biosensors in which the GQDs serve as fluorescent probes, and the biopolymers provide a
biological function. This work indicates that the integrative self-assembly of biomolecules and
GQDs will open up new avenues for the design of multifunctional biomaterials with combined
optoelectronic properties and biological applications.
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The design of a novel functional peptide molecule was reported by Li et al. [116], which
has the abilities to form peptide nanofibers (PNFs) and recognize with GQDs and GO nanosheet
specifically. Based on the design of peptide sequence, the ternary GQD-PNF-GO nanohybrids
are successfully synthesized. It is found that the GQDPNF-GO nanohybrids show potential
applications for high performance electrochemical hydrogen peroxide (H2 O2 ) biosensor. This
fabricated biosensor exhibits high sensitivity and selectivity, low detection limit, and wide linear
range for sensing H2 O2 . The strategies shown in this work will benefit the further creation of
functional binary and ternary nanomaterials, as well as the understanding of their self-assembly
and formation mechanisms.
Significantly, Ju et al. [117] synthesized highly blueluminescent nitrogen-doped
graphene quantum dots (N-GQDs) by a facile one-step hydrothermal treatment of citric acid and
dicyandiamide. A quantum yield (QY) as high as 32.4% is achieved at an excitation wavelength
of 350 nm. It is found that such N-GQDs with a high QY can be used as efficient fluorescent
probes for the detection of glutathione (GSH). In the detection, the photoluminescence (PL)
intensity of the N-GQDs could be quenched by mercuric ions due to the strong electrostatic
interaction and electron transfer between N-GQDs and Hg(II). Upon the addition of GSH, the
PL intensity of N-GQDs can be recovered owing to the preferred combination of Hg(II) and
GSH by forming a Hg(II)-S bond. Under optimal conditions, this fluorescence turn-on sensing
system exhibits excellent sensitivity and selectivity for GSH determination with a detection limit
of 87 nM. Importantly, the N-GQDsHg(II) system can be successfully applied for visualizing the
intracellular GSH in live HeLa cells due to bright luminescence, low cytotoxicity, and good
biocompatibility. Furthermore, they designed a novel colorimetric sensor for more sensitive
detection of GSH based on the intrinsic peroxidase-like activity of Ag nanoparticles supported
on the nitrogen-doped graphene quantum dots (Ag NPs-NGQDs) [118]. The Ag NPs-N-GQDs
nanocomposites can catalyze the oxidation of 3,3′5,5′-tetramethylbenzidine (TMB) to oxTMB
with the presence of H2 O2 along with a visual color variation. As GSH can reduce oxTMB, the
UV-vis absorption intensity of oxTMB shows a strong dependence on the concentration of GSH.
The novel Ag NPs-NGQDs-TMB-H2 O2 system exhibits more high sensitivity and selectivity
for the detection of GSH with a lower detection limit of 31 nM and a wide detection linear range
of 0.1–157.6 μM. This novel sensor system shows great potential application for GSH detection
in blood serum sample and easyto-make analytical approaches in the future.
ISBN: 978-81-948129-1-3 Page 220
3.2 BIO-IMAGING
GQDs have a potential application in the field of bio-imaging because of their excellent
fluorescence properties and the low cytotoxicity. GQDs were applied in cell imaging of human
breast cancer cell line MCF-7 by Dong et al. [59]. The work shows that the cell membrane,
cytoplasm, and nucleus can be marked with fluorescence by GQDs. It is the first time that the
fluorescent carbon material has been used as fluorescent tags for a cell nucleus.
As we know, there had been no direct effective technology for imaging of stem cells for a long
time, due to the particularity of the stem cells. It is significant that GQDs were used in imaging
of stem cells by Zhang et al. [52] in order to solve this problem. Three different kinds of stem
cells were cultivated with GQDs, and the results showed that GQDs can smoothly enter into the
stem cells. It was found that GQDs were not observed in the nucleus, indicating that it will not
cause genetic disruption of the stem cells. It showed that GQDs had relatively low cytotoxicity.
In contrast, when stem cells were mixed with CdS quantum dots, it soon died, due to the toxicity
of the heavy metals Cd2+ . This approach demonstrates the distinctive advantages of GQDs for
direct and efficient stemcell labeling, opening up great opportunities for their biomedical
applications.
Wang et al. [119] reported the first realization of the industrial-scale (20 l) production of
high-quality fluorescent GQDs via a molecular fusion route from a low-cost, active derivative of
pyrene. The as-prepared GQDs show superior optical properties including strong excitonic
absorption bands extending to ~530 nm, bright PL at 510 nm with a quantum yield of up to
42%, and a wide PL spectrum. The GQDs are applied as biological fluorescent probes for
visualizing and targeting the Golgi apparatus in HeLa and MCF7 live cells. The low-cost mass
production, excellent biocompatibility, and superior optical properties make the GQDs an
attractive alternative probe for efficient Golgi-targeted imaging in biomedical applications.
Ge et al. [120] presented a new photodynamic therapy (PDT) agent based on GQDs that
can produce singlet oxygen (1O2) via a multistate sensitization process, resulting in a quantum
yield of 1.3, the highest reported for PDT agents. The GQDs also exhibit a broad absorption
band spanning the UV region and the entire visible region and a strong deep-red emission.
Through in vitro and in vivo studies, they demonstrate that GQDs can be used as PDT agents,
simultaneously allowing imaging and providing a highly efficient cancer therapy. The present
work may lead to a new generation of carbon-based nanomaterial PDT agents with overall
ISBN: 978-81-948129-1-3 Page 221
performance superior to conventional agents in terms of 1O2 quantum yield, water dispersibility,
photo- and pH-stability, and biocompatibility.
In the work of Su et al. [121], the design of a peptide with trifunctional motifs is reported
as the precursor building block for constructing a novel multifunctional protein nanofiber (PNF)
and further conjugated with highly fluorescent GQDs by noncovalent interactions. The GQDs
essentially maintain their favorable optical properties in the PNF-GQD nanohybrids. A good
biocompatibility of the PNF-GQD nanohybrids is found with cell viability assays. With both a
recognition moiety (RGD) and an imaging probe (GQD), these PNF-GQD nanohybrids possess
the capability of targeting and imaging tumor cells simultaneously. This research demonstrates
that GQD-decorated PNF nanohybrids have great potential as multifunctional platforms for
biomedical applications, particularly, where the capability of sensitive tracking and efficient
labeling is appreciated.
3.3 DRUG DELIVERY
There have been reports that graphene or graphene-based nanomaterials are utilized as
carriers for delivery of drugsto improve the delivery efficiency or offer benefits in the therapeutic
effect [122, 123]. Compared to graphene, GQDs have the better water solubility, lower
cytotoxicity, and larger specific surface area, and also can combine with a variety of compounds
through the intermolecular π-π interaction. Therefore, GQDs are the more effective carriers or
loaders of drug molecules. For instance, a biocompatible and cell traceable drug delivery system
based on GQD, for the targeted delivery of the DNA-intercalating drug doxorubicin (DOX) to
cancer cells, was reported by Iannazzo et al. [124]. The GQDs were covalently linked to the
tumor-targeting module biotin (vitamin K or vitamin B7), able to efficiently recognize biotin
receptors overexpressed on cancer cells by means of a strategically designed cleavable linker,
which can be specifically activated inside cells. The therapeutic agent used in this study, DOX,
was loaded to the GQD surface, taking advantage of the excellent absorption properties of
carbon nanomaterials, by π-π interaction. Moreover, the inherent fluorescence allowed the drug
release to be tracked (Figure 19). This study has designed a new biocompatible and cell-traceable
drug delivery system, able to release the therapeutic agent to cancer cells in a selective manner
and minimize the anticancer drug systemic toxicity and undesirable side effects, typically
associated with conventional chemotherapy.
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In the study of Ghanbari et al. [125], blue fluorescent nitrogen-doped GQDs (N-GQDs)
were synthesized by a hydrothermal method via pyrolysis of citric acid as the carbon source and
urea as the nitrogen source. For the first time, the N-GQDs have been loaded with the anticancer
drug, methotrexate (MTX), to prepare the MTX-(N-GQDs) as an efficient drug delivery system.
The in vitro cytotoxicity of the MTX-(N-GQDs) on human breast cancer cells investigated
suggests that the drug-free N-GQDs nanocarriers are highly biocompatible, whereas the MTX-
loaded ones are more cytotoxic than the free MTX. This research confirms the accomplishment
of GQDs as nanocarriers to prolong the cytotoxic effects of its loaded drug for the better killing
of cancer cells.
Significantly, a size-changeable GQD nanoaircraft (SCNA) that served as a hierarchical
tumor-targeting agent with a high cargo payload was developed to penetrate and deliver an
anticancer drug into deep tumors [126]. The nanoaircraft is composed of ultrasmall GQDs (less
than 5 nm) functionalized with a pH-sensitive polymer that demonstrates an aggregation
transition at a weak acidity of the tumor environment but is stable at physiological pH with
stealth function. A size conversion of the SCNA at the tumor site is further actuated by near-
infrared irradiation, which disassembles 150 nm of SCNA into 5 nm of DOX/GQD like a bomb-
loaded jet, facilitating the penetration into the deep tumor tissue. At the tumor, the penetrated
DOX/GQD can infect neighboring cancer cells for repeated cell killing.
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Figure 19: GQD for tumor-targeted DOX delivery (reprinted/reproduced with the permission of
Ref. [124], copyright 2017, Elsevier B.V.)
Such an SCNA integrated with combinational therapy successfully suppresses xenograft
tumors in 18 days without distal harm. This work presents a sophisticated strategy, which
displays the hierarchically targeted and penetrated delivery of drugs and energy to deep tumor
and shows potential for use in other tumor therapies.
3.4 SOLAR CELLS
Solar cells are one of the hot research topics in the field of clean energy. The major factor
for the use of GQDs in solar cells is the size-dependent bandgap tuning property. The GQDs
have very attractive benefits for applications in solar cells compared with other materials such as
silicon and perovskite, mainly because of better dispersion, the band gap which can be regulated,
high chemical stability, and low toxicity. Additionally, GQDs have the quantum confinement
effect and edge effect, well suited to solar cells. To date, many kinds of GQD-based solar cells
have been reported, such as the use of hole transport layer (HTL) material [127], silicon/GQD
heterojunction solar cell [128], semiconductor/GQD solar cell [129], and conductive polymer-
doped GQD solar cell [130]. Most researches have focused on the use of GQDs as an electron-
hole transfer material and as a tuner of the band gap.
ISBN: 978-81-948129-1-3 Page 224
In the work of Li et al. [131] derived from doublewalled CNTs, GQDs with a uniform
size distribution were prepared through solution chemistry. The GQDs in chlorobenzene exhibit
bright blue emission upon UV excitation. The introduction of the GQDs into a bulk
heterojunction polymer solar cell (PSC) based on poly(3- hexylthiophene):(6,6)-phenyl-C61
butyric acid methyl ester (P3HT:PCBM) results in a significant enhancement of the power
conversion efficiency (PCE). The efficiency can be further improved by adjusting the PCBM
content in the active layer, reaching a maximum PCE of 5.24%. This ternary system based on
blended P3HT:PCBM:GQDs represents a new method to enhance the efficiency of PSC. This
work expands the application of GQDs to PSC devices.
Kundu et al. [132] reported an enhanced PCE of 11.7% ± 0.2 and a fill factor (FF) of
71% for dye-sensitized solar cells (DSSC) with an active area of 0.16 cm2 after modifying the
TiO2 photoanode with size-selective (ca. 2 nm) N,F,S-codoped GQDs (NFS-GQDs), which
exhibit a PL quantum yield of 70%. An upward shift in the Fermi level has been observed,
perhaps responsible for the improved performance along with the possibility of preventing the
back electron transfer from TiO2 . This work indicates that the incorporation of size controlled,
hetero atom-doped GQDs can enhance the efficiency of DSSCs, enabling more opto-electronic
applications.
Besides the above applications, research has progressed step by step until now, and the
GQDs are applied in materials or various complex devices, such as catalysts [133, 134],
photovoltaic devices [135, 136], LEDs [137], lithium ion batteries [138], supercapacitors [139,
140], photodetectors [141, 142], tissue-engineering materials [143], etc.
4 CONCLUSIONS AND OUTLOOK
As discussed in this article, many kinds of preparation methods of GQDs are developed.
The basic concept and classical method for making GQDs is cleavage of carbonaceous materials
by repeated oxidation and reduction. Besides, there are other ways including controllable
synthesis and carbonization. At present, the hydrothermal process using GO as a starting material
to obtain GQDs is the environmentally friendly approach, and strong-acidic treatment of graphite
is the method which can achieve mass production. For further development of the GQDs, we
need to find the commercially available and effective methods to fabricate various GQDs.
Devices and applications can benefit from the GQDs because of their special optical and
electrical properties. Promising strategies have been provided by elaborate designs of micro-
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structures and device structures for practical applications and future researches. However, the
research on the GQDs is still in its early stage compared to graphene. There is still a long way to
go for extensive practical applications, and there is also a wide space of exploration for
researchers.
First, there are still many issues waiting to be settled for further development. For
instance, it is still a challenge to find scaled but facile synthetic methods for the production of
GQDs with high quality, though much research on the fabrication methods has been published.
Furthermore, conclusive evidence and convincing explanation for the PL mechanism of GQDs
are still absent. In addition, it is still unclear how the factors, such as size, doping, crystallinity,
and surface functionalization, have influence on the optical properties. Moreover, both indepth
experimental verification and theoretical calculations are much desired. For example, a catalytic
model should provide a conceptual framework to help understand how GQDs and active
components interact. It is also important to determine the types of defects or sites that are most
appropriate for tuning the interactions and how this can affect the reaction mechanism. All of the
issues inhibit the development of GQDs.
Additionally, further researches for potential applications are also important and must be
paid more attention, besides the fundamental studies. For application in bio-imaging in vivo and
in vitro, long-wavelength emission or up-conversion luminescence is more appropriate for the
biological window. However, quantum yield of GQDs is often low when excited with long
wavelength light. Furthermore, most of the GQDs exhibit bright blue to green luminescene, but
high efficient long-wavelength emission is still absent to obtain WLEDs with high color
rendering index (CRI), as well as warm white light. It is suggested that large sp2 domain is
favorable for yellow and red emission, but the method is of low efficiency [144]. Therefore, new
methods for large-scale production of GQDs with long emission wavelengths are expected.
Additionally, though GQDs are expected to play an important role in novel devices, such as
photovoltaic devices, the synthetic variability hinders reproducibility and affects the efficiency.
The size and surface molecules need to be fine tuned if efficient devices are to be prepared. All
of these issues are waiting to be settled for further development. We hope further researches can
successfully solve the problems in the near future. Under the joint efforts of researchers, we
believe that not only novel preparation methods and properties of GQDs will be found, but also
new devices and applications will be invented.
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ADVANCEMENT OF IN SILICO APPROACHES FOR PROTEIN-PROTEIN

INTERACTION
Sugumari Vallinayagam1 and Kavita Khatana2
1
Department of Biotechnology, Karpaga Vinayaga College of Engineering and Technology
Chengalpattu – 603308, Tamilnadu, Email: sugumari.ias@gmail.com
2
Department of chemistry, School of Basic and Applied Sciences, Galgotias University,
Greater Noida, Email: kavits.khatana_2015@galgotiasuniversity.edu.in
ABSTRACT:
The advent of advanced molecular modeling software, big data analytics, and high-speed
processing units has led to the exponential evolution of modern drug discovery and better
insights into complex biological processes and disease networks. This has progressively steered
current research interests to understanding protein-protein interaction (PPI) systems that are
related to a number of relevant diseases, such as cancer, neurological illnesses, metabolic
disorders, etc. However, targeting PPIs are challenging due to their “undruggable” binding
interfaces. In this review, we focus on the current obstacles that impede PPI drug discovery, and
how recent discoveries and advances in in silico approaches can alleviate these barriers to
expedite the search for potential leads, as shown in several exemplary studies. We will also
discuss about currently available information on PPI compounds and systems, along with their
usefulness in molecular modeling. Finally, we conclude by presenting the limits of in silico
application in drug discovery and offer a perspective in the field of computer-aided PPI drug
discovery.
KEYWORDS: protein-protein interaction; peptidomimetics; hot spots; network analysis;
machine learning;
1. INTRODUCTION
Rational drug discovery and design has progressed at a precipitous pace with the aid of in
silico strategies and innovations in hardware and computational power. While still in its infancy
as compared to traditional drug discovery techniques, computer-aided drug discovery (CADD)
has helped deliver several success stories [1–11] that inspired confidence in its continuous
application in the pharmaceutical industry. CADD lends cost- and labor-efficiency in the
identification of potential hits for a therapeutic target before delving into extensive experimental
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assays. The use of computational tools requires the availability of vast amounts of information
on protein and ligand structure, protein function, and an expert grasp of intermolecular forces
and energies necessary for the interaction between binding partners. Its benefits encompass the
different stages of drug discovery and development, including target and hit identification,
structure-activity relationship studies, compound optimization, and analysis and prediction of
lead pharmacokinetic properties. Nevertheless, like any other approaches, in silico methods are
not infallible and are more valuable when they are employed in combination with other drug
discovery tools.
In the last couple of decades, protein-protein interactions (PPIs) have become popular
therapeutic targets. Proteins interact with other biologically important molecules such as
peptides,proteins (homo or hetero), DNA, and RNA to carry out their functions. In general, PPIs
are involved in effecting regulatory changes in response to external stimuli. Different PPI targets
have been implicated in cancer, metabolic diseases, neurological disorders, and many other
diseases [12,13]. The data generated through experimental techniques for these targets are often
hindered in terms of manpower, cost, time, accuracy, and interactome coverage [14]. In light of
this, relying only on experimental techniques may be detrimental to research efforts, as massive
amounts of resources are required to fully explore the human protein interactome. Moreover,
PPIs continue to be difficult targets for drug discovery efforts, due to the differences in PPI
binding site properties and ligand chemical space when compared to traditional protein targets.
As a solution, computational tools have been utilized to fill in the gaps, and to supplement
experimental methods in the investigation of these complex targets.
In this review, we will initially discuss the significance of PPIs as therapeutic targets in
the drug discovery paradigm, and how this has been dealt with so far in the research field. Next,
we will expound on how various in silico methods and recent advances in this field can facilitate
PPI drug discovery, as exemplified in several case studies. Lastly, the benefits and pitfalls of
CADD techniques in relation to PPI drug discovery will be reviewed. Due to the breadth of PPI
drug discovery, we limit the scope of this review to the computer-aided rational discovery and
the development of small molecules, peptides, and peptidomimetics for PPI.
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2. RELEVANCE OF PPISIN THE DRUG DISCOVERY PARADIGM

With an estimated 650,000 PPIs as part of the human interactome [15], it is evident that
these interactions play crucial roles in various cellular processes and pathways. Dysregulation in
PPIs are often found to be the primary cause of several disease pathologies, making them
attractive drug targets. However, the development of PPI inhibitors and stabilizers has been
hindered because of the seemingly low druggability of PPI interfaces. Extensive studies for PPI
targets and modulators have been performed to understand these complex targets and identify
distinct properties in its network, conformational structure, and ligand chemical space. PPI
druggability continues to be poorly understood because these targets show substantial diversity
as compared to protein families that have traditionally been targeted until now. Despite this, its
relevance to multiple diseases and relative novelty in the drug discovery field encourage
researchers to pursue these difficult targets. Moreover, the discovery of PPI inhibitors and
therapeutics in the last few decades have ascertained that these targets are tractable and can be
modulated by small molecule compounds.
2.1. STRUCTURAL FEATURES OF PPIS
PPI interfaces are shallow and highly hydrophobic with large contact surfaces
(>1000 Å2 ) [16,17] that were thought to be involved in their entirety for the formation of protein
complexes. Due to this, PPI targets were portrayed as “intractable.” Later, alanine scanning
experiments revealed the existence of “hot spots,” which are crucial residues in the protein-
protein binding interface that act as chief contributors to the binding free energy. Hot spots
correspond to structurally conserved regions primarily composed of tryptophan, isoleucine,
arginine, and tyrosine, and often form clusters where compounds can potentially bind [18,19].
Though hot spots are frequently bundled closely together in a PPI interface, there are cases
where regions are separated but still work together to allow tight attachment [20]. Aside from the
presence of hot spots, other parameters, including contact surface area, polarity, flatness, and
buriedness, have been employed to characterize PPI interfaces [21–26]. Generally, a PPI
interface is split into a core and a rim region. The core region is buried, and consists of residues
with higher hydrophobicity and conservation, whereas the rim region is in the adjacent solvent-
accessible area with more polar and flexible residues [27–30]. Because the core regions are more
buried and can form interfacial grooves, PPI inhibitors often target these areas and are therefore
usually hydrophobic.
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Different types of PPI complexes are found in biological systems. Obligate complexes
are PPIs that require permanent interaction between protein partners or subunits to establish
function, such as in the cases of the P22 Arc repressor [74] and V-ATPase [75]. In contrast, non-
obligate PPIs have both stable functioning complexes and protomers in vivo [76], like the
GroEL-GroES chaperonine system [77]. Obligate complex interfaces are usually more
hydrophobic than non-obligate complex interfaces, which show higher polarities in order to exist
as independent protomers in solution [17,78]. Protein partners can also bind to each other in a
transient fashion, such as with key interactions involved in cell signaling and regulatory
pathways.
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Weak transient PPIs are commonly formed by disordered proteins or oligomers that have
several states in vivo, leading to interactions that are easily formed or broken based on the
conformation of each protein partner. On the other hand, strong transient complexes require a
molecular trigger, such as ligand binding or posttranslational modification (PTM), to instigate a
change in complex structure [79]. Weak transient complexes are usually mediated by changes in
pH or temperature, and have small and planar surfaces, while those influenced by strong
molecular triggers are often larger and more hydrophobic [80]. PPI interfaces with a surface area
larger than 1000 Å2 are noted to be more flexible and prone to induced-fit conformational
changes [17,80].
PPI complexes display a wide range of affinity and stability. However, high specificity is
still observed, even with proteins that have multiple partners or “protein hubs” [81]. When
dealing with binding specificity of protein hubs, key aspects include molecular recognition and
binding affinity. Orientation and electrostatic complementarity between protein partners are
found to be crucial for identifying the correct protein partner and forming a pre-complex.
Subsequently, short range interactions between partner hot spot regions establish the binding
affinity for the stabilization of the complex [82]. PPI binding interfaces exhibit various means of
mediating specificity of interaction, including changes in conformation, binding site properties,
and the presence of specificitydetermining sites.
While hot spots are considered to be the main contributors for binding affinity and
stability, not all hot spots participate in specificity, as a portion are often shared among several
partners that bind to the same site of a protein hub. Separate specificity-determining hot spots
that can distinguish between cognate and non-cognate partners are present in PPI binding
interfaces [81,83]. The presences of anchor residues and anchoring grooves have been remarked
upon as critical factors for molecular recognition between protein partners [84]. This was
observed in a study done by Kimura et al. [85] wherein mutation of Lys15 of the bovine
pancreatic trypsin inhibitor led to a vast decrease in association rate with trypsin. On the other
hand, mutating Arg17 allowed association with trypsin but resulted in a large increase in the off
rate. These indicate that although both can be classified as hot spots, each residue contributes
distinctive functions in the PPI complex—Lys15 is required for recognition and initial binding,
while Arg17 is critical for the stabilization of the complex [85].
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For facilitated recognition and complex formation, Rajamani and colleagues [84] stated
that anchor residues are usually in their bound-like conformations, while the rest of the hot
regions are more flexible and buried in the unbound state. Aligned with this, molecular dynamics
(MD) simulations of the receptor binding site did not exhibit any significant changes in pocket
conformation or backbone rearrangements, suggesting that the grooves to which the anchor
residues bind are predefined [84]. Interaction with several different partners can also constitute
the presence of multiple binding sites, corresponding to distinct protein subunits, where various
ligand proteins interact to elicit specific responses or functions [86]. Recognition of a specific
protein subunit is influenced by the differences in amino acid (AA) compositions in the binding
interface and the inherent diversity of PPI structural features, even among members of the same
protein family [14,86].
PTM is an alternative process where proteins recognize different protein partners to carry
out their specific functions. A protein can be modified using different PTMs in one or different
residues at the same time, or in a sequential manner, to increase its complexity and functional
scope. Data statistics from dbPTM show that over 60% of PTM sites are situated in protein
functional domains involved in PPIs, implying that PTMs play a crucial role in the modulation of
PPI function [87]. Phosphorylation is one of the most common type of PTMs found in cell
signaling mechanisms and is frequently found on heterooligomeric and transient interfaces. It
can regulate protein function by influencing specific recognition, allosteric regulation, protein
modularity, and binding [88]. This is in agreement with a study conducted by Duan and Walther
[89], where they noted that proteins subject to PTMs, most especially phosphorylation, are
observed to have central roles and a wide interaction range in the human protein interactome
[90].
Structural flexibility can also greatly affect binding affinity and specificity in PPIs. The
dynamic nature of proteins allows them to alter conformations based on the required interactions
for binding with a partner and inducing a corresponding function. Some proteins can subtly
influence both specificity and affinity by altering the orientation of only one or a few conserved
residues in the binding interface, such as MDM2 [91] and proteins with the Src homology 3
(SH3) domain [92,93].
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In other cases, large global or local conformational differences are observed between
different functional states. Conformational adjustment upon protein partner binding can be
explained via two possible mechanisms: induced-fit [94], wherein conformational change occurs
upon interaction of the partners, or population-shift [95], where it is postulated that there exists a
dynamic equilibrium consisting of an ensemble of conformations from which a certain portion
exhibit the conformation a particular partner preferentially binds with [95,96]. This paradigm is
exhibited by intrinsically disordered proteins (IDPs) or regions (IDRs), which have high
structural flexibility and diverse conformations in their native functional state. As opposed to the
conventional theory, where a unique sequence defines a unique three-dimensional (3D) structure
and function [97–99], IDPs and IDRs cover a wide range of conformational space, and have
distinct structural arrangements at a given time point [97,100], enabling promiscuous and
transient interactions with different protein partners. The high modularity of these structures is
linked to crucial roles in cellular processes, and requires tight regulation via changes in
subcellular concentration, diverse conformations, and PTMs [100,101].
For the design of efficient PPI modulators, a great level of understanding of the target is
needed: the type of complex formed, elucidation of binding epitopes and hot spots, determination
of binding site flexibility, etc. While more information is progressively coming to light regarding
PPI binding requirements, further characterization of features that synchronize molecular
recognition, affinity, and specificity within the target protein interface is vital in designing
therapeutics for a specific PPI and function to improve selectivity and off-target toxicity.
2.2. CHARACTERISTICS OF PPI MODULATORS
Increased understanding of druggability and molecular recognition in PPIs has
undoubtedly stimulated drug discovery efforts for these previously intractable targets, offering
hope in identifying small molecule candidates for PPI-regulated pathways. However, the
development of PPI modulators is still at a relatively slow pace in comparison to conventional
small molecule drugs, due to the challenges conferred by the structural optimization required to
improve the affinity and pharmacokinetic properties of the PPI ligands. The complexity of a PPI
inhibitor primarily depends on the interface structure, and this is usually equivalent to that of its
targets. Analysis of known PPI modulators showed that these compounds are larger, more
hydrophobic, and contain more multiple bonds and aromatic rings [102–104].
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Interestingly, PPI ligands have a higher topological polar surface area (TPSA), even with
its high hydrophobicity, as compared to traditional drugs, resulting in their tendency to form
more hydrogen bonds in the PPI surface. This can be attributed to their larger sizes that allow for
the attachment of more polar atoms [103]. More success is acquired for PPIs with grooves or
small pockets, compared to globular interfaces. However, those with hot spot pockets that are
spread out across a large interface are naturally more challenging. Because of this, PPI inhibitors
also tend to have a more 3D conformation than the average inhibitor [105,106], especially for
extended surfaces and disjointed pockets, due to its propensity to mimic the binding epitope of
the partner protein and anchor into small pockets present in PPI interfaces [104,106–108]. Based
on these, it is evident that PPI inhibitors occupy a different chemical space as compared to
typical small molecule inhibitors, and hence, should be carefully evaluated in a different manner
[103,108]. Whereas the primarily hydrophobic interacting features of PPI inhibitors can be
buried in hot spot regions, the rest of their structures are often solvent-exposed due to their
binding location on the protein interface. Molecules 2018, 23, x FOR PEER REVIEW 11 of 43
These elements can be exploited to tailor pharmacokinetic properties without negatively
influencing binding affinity [14]. Studies suggest that 15–40% of the human interactome is
comprised of protein-peptide interactions [109]. Natural peptides have been used as leads for PPI
inhibition [110–112] due to their biocompatibility, low toxicity, and high modularity, which can
aid in both potency and selectivity [113]. However, these ligands tend to have low bioavailability
because of their high propensity for proteolysis, making them poor drug candidates. Several
other approaches are utilized for the discovery, design, and optimization of PPI inhibitors: (a)
using unnatural AAs and other synthetic modifications to the peptide backbone for the design of
peptide mimics (i.e., peptidomimetics) to improve bioactivity and pharmacokinetic properties of
peptide leads; (b) exploiting natural cyclic and macrocyclic peptides that are large enough to
bind to a PPI interface while still possessing favorable properties that can overcome proteolysis
and difficulties in cell permeability; (c) designing and engineering miniproteins via phage
display methods to target large PPI interfaces with high specificity [114,115]; (d) fragment-based
drug design (FBDD) to identify low molecular weight (MW) fragments that targets hot spot
clusters [116,117].
Traditionally, potency was the chief facet used for the early evaluation of lead candidates.
However, it has been established that potency alone does not completely explain bioactivity
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against a target, and other physicochemical properties should also be considered [118]. Advances
in organic and combinatorial chemistry, as well as changes in therapeutic target profiles, resulted
in the growth of both chemical and drug-like spaces over the years [119,120]. Consequently, the
rubrics that were often used to evaluate drug candidates before, have significantly evolved. Since
PPI inhibitors are generally bigger and more hydrophobic than conventional small molecule
drugs, and it is difficult to accurately assess its drug-likeness using typical metrics like Lipinski
[121] and Veber rules [122].
Recent assessment of current drugs gave rise to beyond-rule of 5 (bRO5) characterization
of orally bioavailable compounds [123,124]. Physicochemical properties that were noted by this
novel classification correlate well with typical PPI modulator features. Moreover, ligand
efficiency (LE) and lipophilic ligand efficiency (LLE) are now more commonly used to
determine drug-likeness. LE quantifies the contribution of a molecule’s structure to binding
affinity, whereas LLE estimates a compound drug-likeness by correlating its potency and
lipophilicity. Both measures are used to normalize potency and physicochemical properties to
better assess a series of compounds [125]. The average LE for the PPI inhibitors was found to be
0.23 kcal/mol per heavy atom, while average LLE was 1.32 kcal/mol, both of which are lower
compared to the preferred values of more than 0.3 [126] and 5 [127], respectively. In
comparison, the average LE and LLE of a typical drug is approximately 0.45 and 4.43,
respectively [118]. Aside from these, the correlation of potency with ADMET properties [118]
and physicochemical properties with promiscuity or selectivity [118,127–129] are also currently
being tackled to further expound our understanding of drug-likeness in relation to compound
activity.
3. EMERGING IN SILICO APPROACHES FOR PPI DRUG DISCOVERY
Experimental methods are widely accepted as the standard for any analysis as they
illustrate biological scenarios in either in vitro or in vivo systems. However, the immensity of the
human interactome requires considerable cost, effort, and time to study. Moreover, PPI studies
are highly dependent on dynamics, PTMs, and physiological conditions, causing difficulties in
distinguishing true interactions from experimental artifacts and inconsistencies in findings,
especially in cases involving transient interactions and IDPs. In silico methods have emerged as
alternative methods or as complements to experimental techniques, to fill in the g
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3.1. DISCERNING THE PPI NETWORK TOPOLOGY

PPI targets have become a class of their own throughout the years. In contrast to well-
known protein families, like GPCRs and kinases, this class of targets encompasses exceedingly
diverse structures and interactions. Efforts have been made to compile information about
experimentallydetermined PPIs into databases or platforms. These repositories, which can
provide invaluable information regarding functional characterization of proteins to ascertain and
weigh their roles in diseases. Primary databases report only experimentally verified protein
interactions from individually published studies that were manually curated to provide
researchers with accessible information. Naturally, curating PPI information is no easy task;
errors and noise are often found in these repositories. To ensure the robustness of study models,
careful data selection must be observed by establishing replication with different methods,
interaction type, subcellular location, and other physiological aspects [130,131]. Access to such
extensive datasets are provided by meta-databases, which report experimentally validated PPIs
collected from multiple primary databases and integrate them into one large data model.
Network analysis and sequence-based methods are valuable strategies that have
widespread application in various stages of drug discovery, including target identification,
binding site prediction, and polypharmacological studies [165]. Due to the impact of PPIs as
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therapeutic targets for diseases of interest, it is beneficial to understand the protein interactome
and its correlation with disease onset and development via comprehensive mapping of the PPI
network (PPIN) topology. Studying network topologies allows for the identification of novel
biomarkers and relevant disease targets that would have taken a long time to discover and
validate using conventional means. Subsequently, sequence and conservation information should
be studied at length to determine their importance to protein dynamics and binding, further
aiding in the rational design of drug candidates. The cornucopia of experimental data that is
available due to advances in structural biology and biophysical methods can be expended for
PPIN analysis.
PPINs exhibit typical web-like structures, wherein most proteins (nodes) display
interactions (edge) with only one or a few partners, while some proteins exhibit contact with
multiple partners, and are hence called hubs [166]. The correlation between topology and protein
function is often measured through node degree and betweenness centrality [167–169]. Protein
hubs exhibit a high node degree, i.e., the number of edges linked to a node, through multiple
associations, rendering them as prominent elements that can affect the network function [170].
Indeed, studies have suggested that protein hubs are highly essential for cellular function, and
can greatly affect the whole network when dysfunctional, making them prime drug targets [171–
174]. On the other hand, proteins that display high betweenness centrality are considered as
bottlenecks, and they work by regulating focal points of communication within the network
[170]. Targeting bottlenecks, as compared to hubs can allow careful control of the interaction
network due to their apparent impact on the strength and integrity of the whole network
[175,176]. Discovery of these network components have already been established as a strategic
source of novel protein targets [177–179].
In a recent study done by Vinayagam and colleagues [179], they performed network
controllability analysis using 6339 proteins involved in 64,814 PPIs to sort proteins (nodes) as
indispensable, neutral, or dispensable for the directed protein interaction network. Aside from
classifying proteins, they observed that indispensable proteins are often conserved among species
and enriched in signaling proteins such as kinases, while neutral proteins are often found as
membrane and transcription factor proteins. Indispensable proteins were also implicated in
genetic and human viral diseases, along with cancer genes. More specifically, they identified 56
genes that are most often deleted or over-expressed in nine types of cancer, with 10 genes
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already under investigation and 46 being proposed as new potential cancer targets. Remarkably,
while indispensable nodes greatly populate current Food and Drug Administration (FDA)-
approved drug targets, they found that these proteins are inadequately represented in the
annotated druggable genome. They postulated that network controllability analysis would be able
to aid in validating the druggable genome, as well as identify novel therapeutic targets [179].
Control theory and network analysis have been applied in other similar studies to identify
essential nodes for cancer [180] and viral infection [181].
PPI prediction is essential for functional annotation. Constraints in experimental
approaches necessitated the application of in silico methods, such as binding site prediction
(Section 3.3) and protein-protein docking (Section 3.4). However, both these methods require
prior knowledge of PPI partners and the availability of 3D structural data. In cases concerning
unknown PPI partners and/or structures, sequence- and coevolution-based methods can be
utilized to predict the probability of interaction and residues pertinent to binding. Multiple
sequence alignment (MSA) of homologs offer knowledge about residue coevolution, which can
predict protein partners based on the premise that partners usually coevolve to maintain their
function [170,182]. Information obtained from these types of analyses can also facilitate the 3D
structure prediction of applicable PPI complexes [183]. Another approach takes advantage of
orthologous associations by predicting PPIs based on interologs, wherein proteins that are
observed to interact in one species, are predicted to also interact in other species if they are found
to be conserved [184]. Several machine learning (ML) models are also based on sequence
information, and these have been highlighted in Section 3.2.
3.2. HARNESSING THE POWER OF MACHINE LEARNING ALGORITHMS FOR
PPIS
To extend the concept of in silico models, intelligent and contemporary ML algorithms
have been implemented in the identification of PPIs. ML is a data-driven or knowledge-based
approach which requires an adequate number of training sets and features. Statistical and ML
methods have been applied at varied stages: assimilation of diverse heterogenous datasets,
assessment of predictions, prediction of prospective PPIs, and investigation of extrapolated PPI
networks [185–188].
During the past, several ML algorithms (i.e., κ-nearest neighbor (κNN), naïve Bayesian
[188,189], neural networks (NN) [190], random forest (RF), support vector machines (SVM),
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decision trees, logistic regression, etc.) have been applied to predict PPIs. ML methods utilize a
dataset of experimentally validated PPI surfaces to train interface predictors and further employ
the trained model for prediction of protein-protein interfacial residues of query proteins [191].
Prevailing ML predictors apply binary classification, where a target residue is classified as either
interfacial or non- interfacial by utilizing the target features or adjacent residues to formulate
predictions [191]. Moreover, the accuracy of the prediction model is dependent on the input
features used for training. Hence, it is crucial to identify the various protein features that are
essential for training a ML algorithm. Several protein features are utilized in the development of
models for PPI predictions, either individually or in combination. However, a single feature does
not support sufficient data in the prediction of PPIs.
Therefore, a combination of features is essential to enhance the performance of ML
methods in PPI prediction. Some of the protein features utilized in model development include
AA types, co-essentiality data, evolutionary information, GO-driven frequency-based similarity
and semantic similarity, MIPS functional catalogue (FunCat), physicochemical properties of
AAs, position-specific scoring matrices (PSSMs), protein expression data, residue interface
propensity, secondary and tertiary structural information, sequence entropy, surface shape, and
solvent accessible surface area [192,193]. Once the model is properly trained with the input
features, its performance is assessed with an external or test dataset. The five main steps
involved in PPI predictions are depicted in Figure 2. In PPI-based ML approaches, the input of
prediction models is in the form of sequence or structural features or both. However, most of the
existing ML interface predictors are structure-based [191]. The merits and demerits of structure-
and sequence-based methods have been well reviewed in [191]. Additionally, there are a few
meta-based approaches, where raw scores from several prediction servers are integrated and re-
computed to improve the prediction performance. Representative structure-, sequence-, and
meta-based ML predictors for identification of PPIs are summarized.
Jones and Thornton developed a method for the prediction of PPI interaction sites by
analyzing a group of residue spots on the protein interface. Each residue patch was examined
using six parameters, including the accessible surface area, hydrophobicity, planarity, protrusion,
residue interface propensity, and the solvation potential. The relative combined score was
calculated to predict the probability of surface patch formation in the PPIs [194]. Furthermore,
the distribution of the observed patch rankings relative to all other surface patches were
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evaluated for each dataset [195]. Likewise, Bradford et al. developed SVM- [196] and Bayesian-
based [197] classifiers using surface patches. Another group developed a PPI predictor, known
as SPPIDER [198], using SVM and NN in combination with relative solvent accessibility (RSA).
The authors showed that the implemented RSA-based features demonstrated superior
performance to other PPI-based features. Ofran and Rost developed a NN-based method, known
as ISIS [190], for the prediction of protein interacting residues from sequence. This ML approach
combines AA composition of protein-protein interfaces, along with evolutionary profiles, solvent
accessibility, and protein secondary structural features. A naïve Bayes classifier, PSIVER, was
developed by Murakami and Mizuguchi for the prediction of PPIs using sequence-based
features, such as PSSM and predicted accessibility. The conditional probabilities of each AA
feature were estimated using a kernel density estimation (KDE) method [199]. Two sequence-
based PPI predictors, namely LORIS [200] and SPRINGS [201], were developed by applying
L1-regularized logistic regression and artificial neural network (ANN) methods based on several
seBesides supervised ML algorithms, unsupervised ML approaches were also implemented in
PPI prediction. Deep learning has become a new dimension of ML field, which attempts to learn
multiple layered models of inputs, known as NNs.
A deep learning algorithm is capable of handling extensive raw and complex data where
it operates by mimicking deep neural networks (DNNs) and learning processes of the human
brain. The central idea of deep learning has been detailed in [202]. It has potent applications in
decision making, natural language understanding, and image and speech recognition. This
algorithm has also been applied in the field of bioinformatics and biopharma industry. Recently,
Sun et al. implemented deep learning for sequence-based prediction of human PPIs [203]. They
applied a stacked autoencoder (SAE) to study PPIs in humans and other species (E. coli,
Drosophila, and C. elegans). The models developed using an autocovariance (AC) coding
method showed the top results on 10-fold cross validation and varied external datasets ranging
from 87.99% to 99.21% (prediction accuracy). Another group developed a method known as
DeepPPI, which applies DNNs to obtain high-level discriminative characteristics from common
protein descriptors sequential features.
Besides supervised ML algorithms, unsupervised ML approaches were also implemented
in PPI prediction. Deep learning has become a new dimension of ML field, which attempts to
learn multiple layered models of inputs, known as NNs. A deep learning algorithm is capable of
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handling extensive raw and complex data where it operates by mimicking deep neural networks
(DNNs) and learning processes of the human brain. The central idea of deep learning has been
detailed in [202]. It has potent applications in decision making, natural language understanding,
and image and speech recognition. This algorithm has also been applied in the field of
bioinformatics and biopharma industry. Recently, Sun et al. implemented deep learning for
sequence-based prediction of human PPIs [203]. They applied a stacked autoencoder (SAE) to
study PPIs in humans and other species (E. coli, Drosophila, and C. elegans). The models
developed using an autocovariance (AC) coding method showed the top results on 10-fold cross
validation and varied external datasets ranging from 87.99% to 99.21% (prediction accuracy).
Another group developed a method known as DeepPPI, which applies DNNs to obtain high-level
discriminative characteristics from common protein descriptors[204]. This method showed better
performance on the external data set, demonstrating an accuracy of 92.50% and a Matthews
Correlation Coefficient (MCC) of 85.08%. and can be used in the development of pertinent ML-
based models.
A decision tree strategy, known as PPI-HitProfiler [232], was built by Reynès and
colleagues based on PPI inhibitors identified in literature, and it is implemented in the FAF (Free
ADME-Tox Filtering tool)-Drugs webserver [233– 236]. In their method, a global
physicochemical depiction of PPI inhibitors was established, along with important descriptors
that relate to the molecular shape (Radial Distribution Function) and the number of multiple
bonds (unsaturation index), that can be used in the classification of compounds occupying the
PPI chemical space. Using different PPI complexes with ligand and bioassay information, their
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model was able to correctly identify 70% of the validated actives and 52% of the inactives [232].
On the other hand, Hamon et al. built an SVM tool, the 2P2IHunter [237], based on PPI
modulator information taken from 2P2Idb. For the model development, they distinguished
molecular descriptors pertaining to an octanol-water partition coefficient, hydrophilicity, MW,
unsaturation, the number of rings, H-bond donors and acceptors, TPSA, and rotatable bonds, as
being essential for categorizing the potential PPI modulators from screening libraries. Their
SVM model showed a high accuracy (96%) and specificity (96%), but very low sensitivity.
However, it boasted a high enrichment factor of 8, making it an efficient method for filtering out
non-PPI molecules from screening libraries. Thus, the application of ML algorithm in PPI
predictions provides a promising approach for the deeper understanding of the vast network of
protein interactions and their modulators. However, even with the success of ML-based
predictors, prediction accuracy and computational efficiency of developed models can still be
improved further [238].
3.3. ELUCIDATION OF INTERFACE CHARACTERISTICS AND HOT SPOT
CONTRIBUTION IN PPIS
The breakthrough of hot spot residue contribution to PPI binding, in conjunction with
innovations in protein structure determination in the last several years, has greatly helped in
characterizing protein interfaces. Even with current efforts to fully characterize the complete
human interactome using well-known genetic and biochemical techniques, numerous targets are
still without structures, and the sheer number of partner interactions involved leads to difficulties
in fully elucidating all of them. PPIs mainly transpire in conserved regions of protein interfaces,
where hotspots are usually identified, and they can cause conformational changes in the overall
protein structure [239]. However, it is unwise to generalize interface topographies due to the
diversity of interface features between different PPI types. Transient interactions are often found
to be pharmacologically relevant, and they have garnered a great deal of interest, especially in
the development of various prediction methods. However, the dynamic capability and short
duration of transient complex formation makes it more challenging to experimentally
characterize than permanent complexes, resulting in fewer data available for model training and
development [240,241]. Transient interfaces also tend to be less conserved and exhibit different
properties compared to their permanent counterparts, suggesting the need for separate predictive
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strategies. Protein hubs are also difficult to assess, since their binding interfaces are shared by
multiple partners [242].
Alanine scanning mutagenesis (ASM) has been an indispensable tool for the
identification of hot spot residues of different relevant protein targets, but this method has only
been applied in a limited number of PPIs. More to the point, experimental alanine scanning
entails the mutation of several residues in the interface, along with biophysical evaluation of
changes in the binding energy for each mutation, making it a costly approach [243].
Computational alanine scanning is a suitable and quick substitute for hot spot detection. The PPI
complex structure of interest is required to be able to estimate the change in binding energy
(ΔΔG), based on the bound and unbound state of the wild type and mutated proteins [244]. This
method can be combined with MD simulations, as shown first by Massova and Kollman [245].
Modifications were added in the last several years, such as the use of distance-dependent
pair potentials (e.g., DrugScorePPI [246]) and interaction entropy [247] for the computation of
residue-specific free binding energies, to increase the accuracy of calculation and prediction of
hot spots.
Docking methods, like Optimal Docking Area (ODA) [248] and pyDockNIP module of
pyDOCK [249,250], have also been used to determine interface hot spots. ODA evaluates
changes in interfaceenergies based on the residue buriedness upon the binding of protein
partners, and can be applicable in characterizing both obligate and non-obligate interactions
[248]. It has been used to characterize PPI complexes, including those of
metallocarboxypeptidases in complex with proteinaceous inhibitors [251], and UNC-45
chaperone protein in complex with myosin [252], providing beneficial information for drug
discovery efforts. Alternatively, residue normalized-interface propensity (NIP) calculated from
protein-protein docking results can be employed to identify hot spot residues on the protein
surface, as is applied in pyDockNIP [249].
The current rise of the big data era has aidedthe return and further innovation of
knowledgebased techniques, especially in the context of drug discovery. The availability of
sequence and structural information has allowed the exploration and identification of
distinguishing features for the analysis and prediction of hot spots. Sequence-, structural-, and
physicochemical-based features have been used in the development of different data-driven
models and in the characterization of relevant PPIs [253]. Protein sequences can offer
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preliminary description and prediction of potential binding sites for relatively unstudied proteins,
especially in the absence of structural information, because of the noted high conservation of
interface residues [239,254]. In combination with interaction prediction mentioned in Section
3.1, coevolution-based analysis can also predict residues that are critical for binding or, when
combined with statistical physics, residues that have an allosteric or indirect effect on the
complex [255]. Structural homology has also been proposed as a useful metric for interface
characterization, since structures evolve at a much slower pace than sequences [256] and
interface structural space is well-conserved [257]. More commonly, physicochemical and 3D
structural characteristics have been applied in the characterization of PPI hot spots. Previous
analyses have remarked upon the opposing characteristics of the interface, wherein both high
hydrophobicity and high solvent accessibility have been observed. This conflicting observation
has been resolved by the identification of core and rim regions, as mentioned in Section 2.1.
These physicochemical features also correlate well with distinct geometric aspects of PPI
binding sites, such as residue buriedness, side chain protrusion, and surface curvature [258]. Due
to the diversity exhibited by PPIs, it is difficult to rely on any individual feature for binding site
and hot spot prediction, and it has repeatedly been shown that the combination of several
attributes has more success in providing adequate information for PPI surfaces [259].
Knowledge-based PPI binding site and hot spot prediction algorithms are exemplified in tools
like ANCHOR [260], HomPPI [261], KFC2 (Knowledge-based FADE and Contacts) [262–264],
HotPoint [265], FTMAP [266], MINERVA [267], and PredHS [268]. These tools and more are
discussed in extensive detail in other articles [244,253].
MD simulations have also been employed in the identification of druggable PPI pockets,
particularly for flexible and transient systems [269]. A typical case is shown in the BCL-XL
protein, which exhibits drastic conformational differences between its unbound and peptide-
bound structures. While BCL-XL is known to be experimentally druggable [270], its apo-
structure appeared to have a shallow pocket and very low druggability [271]. Analysis of its MD
trajectory revealed that the BCL-XL structure considerably fluctuates, resulting in the formation
of a highly druggable binding region and implication that much of its druggability can be
attributed to its flexible structure [271].
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3.4. EXPLORING THE PPI INTERFACE THROUGH MACROMOLECULAR

DOCKING AND VIRTUAL SCREENING
It has been estimated that the amount of interactions in the human interactome range from
130,000–650,000 [15,272–274]. However, there is a huge gap between the estimated PPIs and
experimentally available structural data. Even though individual protein components are known,
the determination of molecular complexes through experimental techniques is challenging, due
to their transient nature. In the RCSB PDB, only limited structural data is available for protein-
protein or - peptide complexes. Determination of 3D structures of protein-protein or -peptide
complexes is necessary to understand their associated biological functions, to predict mutation
effects, and to aid in the rational design of novel PPI-based drugs. However, due to its associated
experimental difficulties along with incurred high costs and the time in determining complexes,
fast and reliable computational approaches, such as macromolecular docking, is necessitated.
Computational docking is one of the potent in silico tools in structure-based approaches,
which has been broadly utilized for PPI-based drug discovery. This method aims to yield bound
structures with low interaction modes (favorable ones) by sampling a very large number of
possible binding modes and evaluating each conformation using scoring functions [191].
Elucidation of protein-protein or -peptide structural complexes through computational
docking has evolved significantly due to the accumulating data on protein structures and
interactions, improved energy functions, and powerful techniques to accelerate the sampling
process [275]. Protein-protein or -peptide docking consists of two major stages: sampling and
scoring/ranking. In the docking (sampling) stage, several conformations will be generated and,
among those, only the potential ones are sampled. Sampling potential orientations for protein-
protein binding using global or local searches is the first stage in a docking protocol. In a global
search, the protein acting as the receptor is fixed and the other acting as the ligand is moved
around the receptor, where all potential orientations between protein partners in the 3D space are
explored. Though this search is computationally expensive, several docking algorithms such as
Fast-Fourier transform (FFT) have been implemented to reduce docking complexity. Whereas, in
a local search, the local protein features are matched to acquire a suitable complementarily.
Additionally, if experimental data is available, then that information will be integrated in
the sampling stage to improve the prediction accuracy of the docking results. The docking phase
can either be rigid or flexible. In rigid-body docking, there are no changes in the structural
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features. However, in flexible docking, conformational changes in the structures are considered
[170]. The docking stage typically generates thousands of putative complexes. Hence, it is
necessary to sort them and acquire the best docking solutions. In the ranking stage,
conformations sampled from the first stage are rescored and ranked using several scoring
functions. The fundamental principles of protein-protein docking have been well reviewed in
[275]. The docking predictions can be evaluated by calculating the RMSD between the predicted
and original complexes, and by measuring the ratio of the predicted interface residues to native
ones.
Due to the increasing number of docking algorithms developed to predict protein-protein
or -peptide complexes, the Critical Assessment of Predicted Interactions (CAPRI) challenge was
initiated to appraise the performance of different docking protocols [276]. Currently available
protein-protein and -peptide docking tools, respectively.
Several studies have shown the applicability of protein-protein docking in PPI research
[326– 328]. Here, we discuss a recent study where protein-protein docking has been applied for
the prediction of molecular associations between proteins. Variations in protein S-
glutathionylation has been linked to several human diseases. It has been reported that glyoxalase
II (GlxII), an antioxidant glutathione-dependent enzyme, plays a role in the S-glutathionylation
mechanism. However, its mechanistic underpinnings remain unrevealed. In a recent study by
Galeazzi et al. [329], PPIs of GlxII propensity with other cellular proteins, such as malate
dehydrogenase, actin, and glyceraldehyde 3- phosphate dehydrogenase, were explored using a
computational protocol involving protein-protein docking and atomistic MD simulations. The
suitability of the docking programs, including ClusPro, GRAMM-X, HADDOCK, HEX,
PatchDock, SymmDock, and ZDOCK was tested using a set of globular protein oligomers. A
reliable protein-protein docking approach that strongly agrees with the experimental findings
was developed.
The docking method was applied to GlxII-protein complexes for determination of their
association stabilities. Subsequently, the predicted docked complexes were subjected to MD
simulations and Molecular Mechanics Poisson Boltzmann Surface Area (MM/PBSA) analysis.
In silico results showed that GlxII highly interacted with actin and MDH. Therefore, this study
highlighted the application of protein-protein docking approaches in the elucidation of the
molecular mechanisms and thermodynamics of GlxII protein binding affinity.
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Similarly, once druggability and potential binding sites in the protein interface have
beenestablished, virtual screening (VS) experiments can be performed to find potential binders
that can disrupt or stabilize the target PPI. VS can be categorized as structure-based (SBVS) or
ligand-based (LBVS), but the former is more useful for PPIs, since ligand information for these
targets is sparse when compared to other drug targets. SB pharmacophore VS can be employed
for a given target, wherein interface features are explored to generate a 3D pharmacophore
model used to identify ligands with diverse scaffolds that complement the binding site and have
the potential to exhibit a desired bioactivity (inhibition or stabilization) [330].
Some programs that employ the pharmacophore strategy include Catalyst [331], FLAP
(Fingerprints for Ligands And Proteins) [332], HS-Pharm (HotSpots-guided receptor-based
Pharmacophores) [333], LigandScout [334], PHASE [335], and AnchorQuery [336].
Alternatively, docking is a well-established method for SBVS, wherein ligand binding is
energetically evaluated by scoring functions based on its conformation and complementarity
with the binding pocket [337]. Scoring functions can be categorized as force field-, knowledge-,
or empirical-based [337]. Each scoring function, in combination with docking programs, has
their own strengths and weaknesses, and selecting a docking program, they should be considered
carefully based on the purpose of the study, i.e., ligand binding prediction (scoring) or VS
(ranking). Examples of popular protein-ligand docking programs include AutoDock [280],
AutoDock Vina [338], GOLD [339], ICM [340], and Glide [341,342].
Here, we present a few case studies where VS and ligand docking protocols have been
incorporated to identify small molecule PPI inhibitors or stabilizers. HIV-1 Nef is involved in
infection, pathogenicity, and disease development by interacting with its own SH3 binding
surface [16]. Betzi et al. [343] combined VS and high-throughput docking to identify small
compounds that can bind to the HIV-1 Nef SH3 surface and prevent HIV-1 Nef/SH3 PPIs.
Initially, the National Cancer Institute (NCI) diversity library was pre-filtered using 14 drug-like
filters, and the identified 1420 compounds were subjected to FlexX docking protocol. The
docked complexes were rescored using GFscore. The top 335 lowest energy compounds were
subsequently subjected to a SH3-based pharmacophoric filter and the resulting 33 potential hits
were clustered. Ten of the molecules were finally selected based on the chemical and
geometrical properties for further experimental validation. Another example focuses on the Bcl-2
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protein family, which are well-established targets for anticancer therapeutics due to their key
roles as apoptotic regulators.
Zhou et al. studied the BCL-XL structure in complex with BCL-2-associated death
promoter (BAD) BH3 peptide, identifying three key hydrophobic features which were used to
create a pharmacophore model. To identify scaffolds with suitable pharmacological and safety
profiles, they screened an in-house database of FDAapproved drugs using the generated
pharmacophore model, identifying three classes of compounds with different cores. From this,
they performed molecular docking on the BCL-XL protein to predictand study the binding
modes of Lipitor and Celexocib, both of which consist of bis-aryl substituted five-membered
heterocyclic scaffold. Both compounds mimicked two out of three hydrophobic features required
for interaction in the BCL-XL binding pocket, leading to the design of BM-501, which showed
suitable affinity to both BCL-2 and BCL-XL and can function as an excellent scaffold in the
design of inhibitors for the Bcl-2 protein family [344]. In a different case, Myc oncoprotein
promotes cancer when it heterodimerizes with Myc-associated factor X (Max), whereas Myc
homodimers suppress cancer [345].
Jian et al. utilized the available crystal structure of Max-Max homodimer to identify
small compound stabilizers through VS using AutoDock. The compounds (1668) which were
screened from the NCI diversity dataset were subjected to blind docking using both Max-Max
homodimer and Myc-Max heterodimer structures. Subsequently, the binding sites were analyzed,
and potential compounds which were specific to Max-Max homodimers were identified. These
case studies are clear indications that structure-based computational approaches can be used for
the discovery of small molecule modulators of PPIs.
3.5. EXPLOITING HOT SPOT REGIONS FOR FRAGMENT-BASED DESIGN
FBDD has been widely employed for different disease targets in the last several years to
acquire new chemical entities, while efficiently scouring a much larger chemical space than
traditional highthroughput screening (HTS) can cover. Compared to extensive compound
libraries commonly used for VS, which comprise of millions of complex structures, fragment
libraries often contain only hundreds to a few thousand low-MW structures [346]. Despite this,
fragment screening reportedly displays higher hit rates and better LE than HTS efforts [346,347].
However, most of the current fragment libraries consist of mostly flat, aromatic structures, which
can still limit the exploration of chemical space. Renewed interest for natural products due to
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their structural diversity and relatively safe pharmacological profiles steered the efforts to use
them as a source of highly 3D fragment structures. The presence of sp3-character and
stereogenicity in natural product-derived fragments can increase the chemical space explored for
an FBDD study [348–351].
During optimization, fragment hits can undergo linking or growing strategies. For those
found to bind in adjacent pockets in the protein interface, appropriate linkers can be selected to
connect the two fragments together. On the other hand, fragments can also be grown
synthetically to the neighboring pocket based on binding site analysis and rational design.
Between the two, linking is found to be more difficult, since the best linkers must be identified to
avoid detrimental effects on lead potency and pharmacokinetic properties. While fragment
binders initially show weaker affinity in screens, their smaller size and low complexity make it
easier to manage physicochemical properties during the optimization than a high-affinity HTS
lead. Nevertheless, these weak binding fragments are discovered to form excellent interactions in
the binding site, and thus, they contribute more than half of the favorable binding energy if the
interactions are preserved during fragment-to-lead optimization [346].
While the expansive interface of PPIs is not very conducive for small molecule inhibitor
discovery, the presence of multiple hot spot regions spread across the surface suggests the
applicability of fragment-based methods. Distinct fragments can be identified for each hot spot,
and can be later linked or grown into whole molecules with novel structures that are specifically
designed for a particular PPI target [352]. FBDD has been successful in providing potential lead
candidates for PPIs. As a continuation of the same study by Zhou et al. mentioned in the above
section, they used the crystal structure of BCL-XL in complex with ABT-737 and previous assay
results to identify a fragment that could be linked to BM-501. Using part of ABT-737 as a linker,
they were able to rationally design low nanomolar affinity leads for BCL-2 and BCL-XL. They
further employed structure-activity relationship and modeling studies to improve potency and
cellular activities of their compounds [344].
It is important to note that hot spot identification and analysis of the receptor protein are
important elements of fragment-based design of PPI inhibitors. Pertinent fragments must be
screened and bound to essential hot spot clusters, and not to negligible areas in the PPI interface.
Otherwise, while tight binding compounds can be designed from suitable fragments, it may not
produce thedesired outcome (i.e., disruption or stabilization of PPIs). An excellent example for
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this is shown by Geppert and colleagues [353], where they analyzed the nuclear magnetic
resonance (NMR) structures of both the apo and holo structures of IFN-α. Potential binding
cavities were identified using PocketPicker [354], and interface hot spots were predicted using
iPred [355].
Remarkably, hot spot prediction was in good agreement with previous mutation studies
and was able to identify the major interaction groove in the protein interface. Pharmacophore
generation and VS of commercially available fragment-sized compounds was performed,
resulting in the identification of six binders. From these, the highest-ranking compound
displayed good affinity, despite its low molecular weight (279 Da). Probing the ligand protein
hot spots can also facilitate the design of PPI inhibitors. The peptide backbone appropriately
positions hot spot residues to surface pockets, which are primarily composed of hydrophobic
groups, while forming a number of hydrogen bonds in other parts of the interface, such as in the
cases of p53-MDM2 and XIAP-caspase 9 complexes. In this event, it is beneficial both to find
fragments that can bind to the surface hot spot regions, and to find linkers that can fulfill the
hydrogen bond requirements for interaction with the receptor protein [356].
3.6. UNRAVELING THE STRUCTURAL AND FUNCTIONAL ASPECTS OF PPIS
USING MD SIMULATIONS
Despite steady progress in the growth of PPI data, comprehensive understanding of such
complexes and their dynamics remains inadequate. The scarcity of 3D structural data for
proteinprotein or -peptide interacting complexes delays PPI research. Moreover, crystallography
cannot detect the presence of druggable hot spots and transient pockets on protein-protein
interfaces, due to their active motions. MD simulations surpass these limitations by facilitating
in-depth analysis of structural, functional, and dynamic aspects of PPI models. They also provide
valuable insights into PPI mechanisms which could be utilized in the design of PPI modulators.
MD simulation is a commonly applied in silico approach for calculating the time-dependent
motion of biological molecules. The initial step in MD simulation of a PPI is the acquisition of a
starting complex structure, which could be obtained through experiments (X-ray, NMR, or
cryoelectron microscopy (EM)), modeling approaches (homology modeling or protein-protein
docking), or PPI databases.
Once the structural template is available, the next step is to prepare the system and
subject it to atomistic simulations in line with software-dependent simulation procedures. Firstly,
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the system’s initial positions and velocities are fixed. Subsequently, the forces among all atoms
are defined by using a preconfined interaction potential, where time-dependent motions of a
system could be traced by solving classical equations of motions [357]. Key utilities of MD
simulations in PPI exploration include structural investigations of PPIs (e.g., identification of hot
spots, functional mechanisms of complexes, possible binding partners, key interacting features of
complexes, and oligomerization mechanistic insights), design of PPI modulators, elucidation of
macromolecular mechanisms, and refinement of low-resolution structural data (Figure 3). In the
case studies below, we highlight the benefits of MD to PPI investigations.
Dixit et al. investigated the functional mechanism of heat-shock protein (Hsp) 90 using
atomistic MD simulations, along with the modeling of principal correlated motions and energy
landscape analysis [358]. Through this analysis, several key areas in the structure-functional
depiction of controlled interactions and the center of the communication networks were
identified. In another study, Ozdemir et al. unraveled the atomistic interactions of the Rho
GTPases, Cdc42 and Rac1, with the scaffolding protein IQ motif-containing GTPase-activating
protein 2 (IQGAP2) using all-atom MD simulations, site-directed mutagenesis, and Western
blotting [359]. They were able to decipher the underlying mechanism of the different
stoichiometries involved in the binding of Rac1 and Cdc42 to GRD, and IQGAP2 dimerization.
It also provided insights on how Cdc42 and Rac1 mediate actin polymerization in metastasis.
The outcomes of these studies assist in the design of inhibitors and additionally provide
structural, stoichiometric, and working insights into the allosteric control of the complex and
protein machinery dynamics
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Figure 3.Varied applications of molecular dynamics (MD) simulations in PPI research. Using
MD simulations, several aspects of PPIs can be explored, such as gaining insights into their
structural, functional, and mechanistic processes, design of PPI inhibitors or stabilizers, and
refinement of PPI structures with lower resolutions.
MD simulations can also assist in PPI hot spot prediction to provide clues for the design
of potential PPI inhibitors. Survivin protein is found to be over-expressed in several solid
tumors; hence disruption of its interaction with substrate proteins is indispensable for the
treatment of cancer. Sarvagalla et al. [360] provided an exhaustive atomistic detail of the dimer
interface of survivinchromosomal passenger complex (CPC) protein by applying a knowledge-
based model for identification of hot spot residues. Subsequently, extensive MD simulations
were utilized to produce an ensemble of conformations which were further utilized for the
estimation of binding free energies of identified hot spots using MM/PBSA and per-residue
energy decomposition. Survivin and CPC interface hot spots were identified from these analyses.
Finally, a pharmacophore model based on the hot spots was generated and virtually
screened against compound databases for the identification of a potential inhibitor targeting
survivin-CPC interaction. Besides hot spot predictions and functional characterization of PPIs,
MD simulations could be utilized for identification of plausible binding partners. It is
hypothesized that Psalmopeotoxin I (PcFK1) targets subtilisin-like serine protease, PfSUB1. To
investigate this hypothesis, Bastianelli et al. [361] applied bioinformatics analysis, protein-
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protein docking, MD simulations, and free energy calculations on these two proteins. MD
simulations was utilized for refining the docked protein-protein complexes and free energy
calculations. Their computational results were confirmed via experimental testing on PfSUB1
purified and active recombinant enzyme, leading to the validation that PcFK1 indeed targets
PfSUB1 enzymatic activity.
It has been widely accepted that most proteins function as oligomers. However, the
oligomerization process often remains unclear. In such scenarios, MD simulations can assist
experimentalists in unraveling the oligomerization processes. Zhang et al. investigated the
formation of small oligomers in the amyloid fibril-formation process of the peptide GNNQQNY
from the yeast prion-like protein Sup35, using explicit solvent MD simulations. Their results
demonstrated that primarily antiparallel dimer forms, and subsequently novel peptides may
complement the assemblies in parallel arrangement, which is in line with the experimental
microcrystal structure of the amyloid fibril [362]. In another recent study, interactions between
Aβ1–42 oligomers with each of the four models of the full-length Amylin1–37 oligomers were
explored using extensive MD simulations, resulting in the elucidation of the link between type 2
diabetes and Alzheimer’s disease[363]. Thus, MD simulation stands as an indispensable tool to
complement experimental screening techniques in PPI research that dissect PPIs at an atomistic
level, with its high accuracy, vigorous validation of force fields, and the extensive availability of
computational resources for performing large-scale simulations of complex protein systems.
4. PITFALLS OF CADD IN THE DISCOVERY OF PPI INHIBITORS
There is no doubt that computational methods have become one of the cornerstones of
drug discovery, especially with the advances in technology and the increase in experimental data
in the last several years. However, as with any other drug discovery and design methods, CADD
has its own strengths and weaknesses when dealing with a variety of drug targets, including
PPIs.
Prediction and filtering tools are rapid and dependable strategies, particularly for targets
with insufficient information at the start of a drug discovery effort. In the absence of 3D
information, sequence-based methods can provide quick and useful insights about conservation
and evolution information, which may also be critical for ligand design. However, information
that is taken from these techniques might not translate well into 3D systems [253]. In fact,
Keskin et al. remarked that conservation-based analysis is not applicable to transient interactions
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due to its poor conservation across species [170]. Most computational tools are dependent on the
availability of structural information, such as in the case of energy- and knowledge-based
techniques. Energy-based methods are built on the understanding of energy terms in complex
systems.
Due to the intricacy of protein structures, dynamics, and interactions, several
approximations are frequently included in the development of these methods to create a balance
between speed and accuracy; thus, they are more useful as estimates for prioritization rather than
data that is convertible to real scenarios [364]. On the other hand, knowledge-based methods
make use of currently available protein and ligand information, and models created from this
technique are only as good as the data used to create it. For example, certain descriptors are more
predictive for specific PPI types [253]. Furthermore, if a model is based on a particular protein
family or ligand class, it cannot be reliably employed to characterize other protein families or
ligand classes. This becomes a hindrance with respect to the intrinsic flexibility and diversity of
PPIs.
FBDD is notably more efficient than conventional HTS, both in experimental and virtual
form. However, several things are needed before pursuing this strategy. First, structural
information is required for fragment screening, making it an unsuitable method for relatively
unknown targets. Furthermore, information about hot spot regions is crucial, as these are the
focus for any type of screening efforts. Second, binding affinity and orientation of fragments
may differ from that of the complete molecule. Linker groups and attachment points must be
considered conscientiously, as their addition to identified fragment hits can greatly affect both
the potency and pharmacokinetic properties of the lead. Third, and the same as other
computational strategies, virtual fragment hits must be validated. However, validation for these
low affinity structures often requires more sensitive and sophisticated methods and expertise, as
compared to conventional HTS [346].
Perhaps the most notable challenge for many computational methods is the dynamic
nature of proteins, as the inclusion of flexibility is extremely computationally expensive, leading
to the use of rigid protein structures by the majority of CADD modules. This drawback becomes
more prominent in PPIs, whose conspicuously flat interfaces are postulated to be highly flexible.
MD simulations assuage these concerns as it can approximate protein movements in a carefully
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monitored system. Improvements in hardware computational power have also aided increased
interest and utilization of MD for the analysis of PPIs [271,365,366].
The listed shortcomings for each method should not be taken as a dissuasion of their use.
In fact, this is reinforcement that more studies are needed to further improve current
computational tools. Moreover, the strengths of each method can be used to make up for the
weaknesses of the others. The combination of two or more CADD tools, along with the
incorporation of experimental techniques, can result in better insights and success in drug
discovery endeavors.
5. CONCLUSION
PPIs have become prime drug targets due to their integral roles in cell signaling and
regulatory pathways. While increased interest for these targets encouraged comprehensive
studies about PPINs, interface conformations and interactions, and PPI modulators, much is still
needed to completely characterize this vast system and to overcome hindrances associated with
PPI drug discovery. Current experimental techniques have facilitated our growing understanding
of PPIs, resulting in the construction of informative databases containing valuable information
involving the human proteome and interactome. Unfortunately, the immensity of the human
interactome renders experimental methods insufficient, necessitating the use of their
computational counterparts. CADD impacts various stages of PPI drug discovery by assisting in
the characterization of PPIs and its unique chemical space, thereby providing better insights into
the design of PPI modulators. Innovations in computational power and algorithms, along with
the growing knowledge and better parameterization of macromolecular dynamics and energetics
relating to PPIs, have led to notable contributions of in silico methods in various PPI drug
discovery efforts.
Overall, we can expect a remarkable future for PPI drug discovery as both in silico and
experimental methods continue to evolve. While each approach has their own significance in the
field of drug discovery and development, it is important to remember that one is not designed to
outshine the other, but to complement it. The integration of different tools, especially in the study
of diverse drug targets such as PPIs, can enhance our understanding of these targets and aid in
the design of potent and effective PPI modulators.
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AN ECO FRIENDLY CONCEPT OF GREEN COMPUTING

Dr. Sriram E
Department of Business Management, Jayamukhi Institute of Technological Sciences,
Warangal, Telangana
Email: emmadisriram@yahoo.com
ABSTRACT
Green computing is an issue that impacts everyone. And it’s an issue shaped by nearly
everyone in the developed and emerging economies. E-waste threatens the planet as both a
source of toxins and through sheer tonnage. The UN Environment Program estimates the globe
may be producing as much as 50 million tons a year in e-waste. Companies within the computer,
electrical appliance and consumer electronics industries have always accounted success by their
bottom line profits. But in a plainly endangered world, a new greener way of accounting seems
to makes more sense. Actions now must be measured in terms of their impact on people, planet,
and profits rather than just the bottom line. Green Computing is a recent trend towards designing,
building, and operating computer systems to be energy efficient. While programs such as Energy
Star have been around since the early 1990s, recent concerns regarding global climate change
and the energy crisis have led to renewed interest in Green Computing. Thrust of computing was
initially on faster analysis and speedier calculation and solving of mare complex problems. But
in the recent past another focus has got immense importance and that is achievement of energy
efficiency, minimization of power consumption of equipments. It has also given utmost attention
to minimization of e-waste and use of non-toxic materials in preparation of e-equipments. World
leaders have also taken move towards this by following some principles. Now it is the time for
the end user community to follow some rules of thumb to achieve partly the benefit of “Green
Computing”. In India, the implement ability of principle of “Green Computing” is facing a
dilemma due to many socio-economic matters and those are linked to be soughed out to pull
India in the mainstream movement of “Green Computing”.
KEYWORDS: E-waste, Toxic, Non-conventional Energy, Eco-Friendly, SMEs and MNCs
ISBN: 978-81-948129-1-3 Page 302

1. INTRODUCTION
Green computing or green IT, refers to environmentally sustainable computing or IT. In
the article Harnessing Green IT: Principles and Practices, San Murugesan defines the field of
green computing as "the study and practice of designing, manufacturing, using, and disposing of
computers, servers, and associated subsystems—such as monitors, printers, storage devices, and
networking and communications systems—efficiently and effectively with minimal or no impact
on the environment."
We love our computers for all the ways they make our lives (and the world) better -- the
wealth of knowledge (and democratizing force) ofthe Internet, the instantaneous communication,
the sophisticated tools that help us work and create and share. But this modern world's greatest
tool is among our most disposable and resource-heavy items. Performance-wise, computer
design has progressed staggeringly well and astonishingly fast but looking at it from a green
perspective, the work has barely begun. It takes a lot of energy to create, package, store, and
move every 10-20 megabytes of data. Even with energy prices as cheap as they are now, it will
soon cost more to power a computer for four years than it does to buy a new one.
When a computer dies it either rots in a landfill, or children in the developing world end
up wrestling its components apart by hand, melting toxic bits to recover traces of heavy metals.
The goals of green computing are similar to green chemistry; reduce the use of hazardous
materials, maximize energy efficiency during the product's lifetime, and promote the
recyclability or biodegradability of defunct products and factory waste. Research continues into
key areas such as making the use of computers as energy-efficient as possible, and designing
algorithms and systems for efficiency-related computer technologies. Green computing is the
practice of using computing resources efficiently. The goals are to reduce the use of hazardous
materials, maximize energy efficiency during the product's lifetime, and promote recyclability or
biodegradability of defunct products and factory waste.
Such practices include the implementation of energy-efficient central processing units
(CPUs), servers and peripherals as well as reduced resource consumption and proper disposal of
electronic waste (e-waste). In 1992, the U.S. Environmental Protection Agency launched Energy
Star, a voluntary labeling program which is designed to promote and recognize energy-efficiency
in monitors, climate control equipment, and other technologies. This resulted in the widespread
adoption of sleep mode among consumer electronics. The term "green computing" was probably
ISBN: 978-81-948129-1-3 Page 303
coined shortly after the Energy Star program began; there are several USENET posts dating back
to 1992 which use the term in this manner.
2. THE DEMERITS
Performance-wise, computer design has progressed staggeringly well and astonishingly
fast but looking at it from a green perspective, the work is at its epoch. It takes a lot of energy to
create, package, store, and move. Conventionally, manufacturing computers includes the use of
lead, cadmium, mercury, and other toxics in general. Usually, computers can contain 4 to 8
pounds of lead alone, according to green experts.
It's no wonder that computers and other electronics make up two-fifths of all lead in
landfills. To counter this growing pollution threat all over the world due to the growing use of
electronic device in general and computers in particular there is a need to look for a eco-friendly
computer. ―Data center servers use 50 times the energy per square foot as an office [does],‖ says
Mark Bramfitt, principal program manager at PG&E. Data centers are the main reason behind
energy consumption, Energy consumed by data centers in the United States and worldwide
doubled from 2000 to 2005, according to Jonathan Koomey, a consulting professor at Stanford
Universityand staff scientist at Lawrence BerkeleyNational Lab. As a result, some companies are
chasing cheaper data center power. Google is building a data center on Oregon’s Columbia River
to tap hydroelectric power, while Microsoft builds nearby in Washington for the same reason.
Financial services company HSBC is building a data center near Niagara Falls.
To keep servers at the right temperature, companies mainly rely on air-conditioning
equipments. The more powerful the machine, the more cool air needed to keep it from
overheating. By 2005, the energy required to power and cool servers accounted for about 1.2%
of total U.S. electricity consumption, according to a report released in February by staff scientist
Jonathan Koomey of Lawrence Berkeley National Laboratory and sponsored by chip
manufacturer AMD (AMD). According to Gartner by 2010, about half of the Forbes Global 2000
companies will spend more on energy than on hardware such as servers. Energy costs, now about
10% of the average IT budget, could rise to 50% in a matter of years, Kumar says.
Faster processors use more power, because they use too much power and their waste heat
increases temperature for which air conditioning necessary, especially in server farms--between
the computers and the HVAC. The waste heat also causes reliability problems, as CPU's crash
much more often at higher temperatures.
ISBN: 978-81-948129-1-3 Page 304
3. THE DESIGNERS PLAN

Designers plan to make future computer more eco-friendly across its entire life span,
from manufacture to recycling [7]:
a) By replacing petroleum-filled plastic with bioplastics plant-based polymers require less
oil and energy to produce than traditional plastics with a challenge to keep these bioplastic
computers cool so that electronics won't melt them.
b) Landfills can be controlled by making best use of the device by upgrading and
repairing in time with a need to make such processes (i.e., upgradation and repairing) easier and
cheaper.
c) Avoiding the discarding will not only control e-waste out of dumps but also save
energy and materials needed for a whole new computer.
d) Power-sucking displays can be replaced with green light displays made of OLEDs, or
organic light-emitting diodes.
e) Use of toxic materials like lead can be replaced by silver and copper.
f) Making recycling of computers (which is expensive and time consuming at present)
more effective by recycling computer parts separately with a option of reuse or resale.
So far, consumers haven't cared about ecological impact when buying computers, they've
cared only about speed and price. But as Moore's Law marches on and computers commoditize,
consumers will become pickier about being green. Devices use less and less power while
renewable energy gets more and more portable and effective. New green materials are developed
every year, and many toxic ones are already being replaced by them. The greenest computer will
not miraculously fall from the sky one day, it’ll be the product of years of improvements.
4. MATERIALS RECYCLING
Recycling [5] computing equipment can keep harmful materials such as lead, mercury,
and hexavalent chromium out of landfills, and can also replace equipment that otherwise would
need to be manufactured, saving further energy and emissions. Computer systems that have
outlived their particular function can be repurposed, or donated to various charities and non-
profit organizations. However, many charities have recently imposed minimum system
requirements for donated equipment. Additionally, parts from outdated systems may be salvaged
and recycled through certain retail outlets and municipal or private recycling centers. Computing
supplies, such as printer cartridges, paper, and batteries may be recycled as well.
ISBN: 978-81-948129-1-3 Page 305
A drawback to many of these schemes is that computers gathered through recycling

drives are often shipped to developing countries where environmental standards are less strict
than in North America and Europe. The Silicon Valley Toxics Coalition estimates that 80% of
the postconsumer e-waste collected for recycling is shipped abroad to countries such as China
and Pakistan.
Unfortunately, in 2011, the collection rate of e-waste is still very low, even in the most
ecologically advanced countries like France. In this country, e-waste collection is still at a 14 %
annual rate between electronic equipments sold and e-waste collected for 2006 to 2009 [3]. The
recycling of old computers [2] raises an important privacy issue. The old storage devices still
hold private information, such as emails, passwords and credit card numbers, which can be
recovered simply by someone using software that is available freely on the Internet. Deletion of a
file does not actually remove the file from the hard drive. Before recycling a computer, users
should remove the hard drive or hard drives if there is more than one, and physically destroy it or
store it somewhere safe. There is some authorized hardware recycling companies to whom the
computer may be given for recycling, and they typically sign a non-disclosureagreement.
Electronics giants are about to roll out ecofriendly range of computers (like desktops and
laptops) that aim at reducing the ewaste in the environment. [4] They are likely to be free of
hazardous materials such as brominates flame-retardants, PVCs (Polyvinyl Chloride) and heavy
metals such as lead, cadmium and mercury, which are commonly used in computer
manufacturing.
5. CONCLUSION
The field of "green technology‖ encompasses a broad range of subjects from new energy-
generation techniques to the study of advanced materials to be used in our daily life. Green
technology focuses on reducing the environmental impact of industrial processes and innovative
technologies caused by the Earth’s growing population. It has taken upon itself the goal to
provide society’s needs in ways that do not damage or deplete natural resources. Mainly this
means creating fully recyclable products, reducing pollution, proposing alternative technologies
in various fields, and creating a center of economic activity around technologies that benefit
the environment. Nowadays [6] we can see people used computer everywhere and anywhere.
This shows that computer nowadays it compulsory to all people including businessman,
university student, employer government and non government, kids, school and lecturer. So far,
ISBN: 978-81-948129-1-3 Page 306
consumers hardly cared about ecological impact when buying computer but they care only
about speed and price. But now some of people realized that they need to do something that
have to reduce pollution and recycle things. This is because computers were made up from
many chemical and harmful things; this can affect the environment and human healthy. If
computers were broken for sure people will throw it away and burned it but they never realized
once they burned it out it release toxic contaminants into the air. This green computing [1] has
many attentions all over the world with one mission ―go green, save green‖.
REFERENCES
1. http://www.csi-india.org/green computing, accessed during May- July 2008
2. http://www.techno preneur.net/information-desk/sciencetech magazine/ 2007/ nov07/
3. Green Computing.pdf, accessed during May- July 2008
4. http://www.tech-faq.com/green computing.shtml, accessed during May July 2008
5. Srivastava, M (2007) Chemistry for Green Environment
6. http://en.wikipedia.org/wiki/Green_compu ting
7. http://www.oppapers.com/essays/Green Computing/577405
8. .http://www.techno preneur.net/information-desk/sciencetech magazine/ 2007/ nov07/
Green%20Computing.pdf
9. http://green-compute.com/
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MAGNETO-ELECTRONIC PROPERTIES OF GRAPHENE NANOPARTICLES

Sridhar Arelli1 and Akshay.S2
1
Department of Physics, OPJS UNIVERSITY, Churu, Rajasthan.
Email: usri89760@gmail.com
2
Department of Physics, Maharani Lakshmi Ammanni College for Women Autonomous,
Malleswaram, Bengaluru-560012, Karnataka
Email: akshay@mlacw.edu.in
ABSTRACT
The effect due to doping by B, Si, N on the magneto-transport properties of graphene is
investigated using the generalized tight-binding model in conjunction with the Kubo formula.
The crucial electronic and transport properties are greatly diversified by the type of dopant and
doping concentration. The contribution from the guest atoms may open a band gap, thereby
giving rise to the rich Landau level energy spectra and consequently the unique quantum Hall
conductivity. The Fermi energy-dependent quantum Hall effect appears as a step structure
having both integer and half-integer plateaus. Doping leads to the occurrence of zero
conductivity, unlike the plateau sequence for pristine graphene. The predicted dopant- and
concentration-enriched quantum Hall effect for doped graphene should provide useful
information for magneto-transport measurements and possible technological applications as well
as metrology.
1. INTRODUCTION
Understanding the physics behind the quantum Hall effect (QHE) has led to a prolonged
effort by numerous theoretical and experimental researchers. In particular, the Hall conductivity
of condensed matter systems under an external magnetic field has received special attention.1–20
Individual behaviors of magnetic-field-quantized Landau levels (LLs) have been employed to
explain the QHE. The electrons filling LLs and wave functions have been demonstrated to be
associated with the integer and fractional Quantum Hall States. In this work, we will
systematically explore the QHE in Si-, B-, and N-doped graphene for various doping
concentrations with the aid of the tight-binding model to determine the energy bands.
Ever since the time when graphene was experimentally discovered, its exotic QHE has
attracted many studies from both the experimental and theoretical points of view.1–14
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Monolayer graphene shows amazing physical properties because of its single-atom thickness and
hexagonal symmetry. The magnetically quantized LLs of the isotropic Dirac cone form a
distinctive energy spectrum for which conduction and valence energy levels are dependent on
both the field strength and the quantum number of specified LLs. Such an established relation, √
Bznc,v, has been verified through experiment using scanning tunneling spectroscopy (STS),21
optical spectroscopies,22 and transport equipment.5,6 Interestingly, it has been realized that the
inter-LL transitions obey a specific selection rule associated with the equivalence of the two
sublattices in the crystal structure. Only excitations between the valence and conduction LLs
whose quantum numbers satisfy ∆ n = |n v − n c | = 1 are allowed, resulting in the exotic QHE.
Magnetic transport measurements have verified the unconventional half-integer Hall
conductivity σxy = (m+ 1/2)4e 2/h (m is an integer and the factor of 4 represents the spin and
sublattice-dependent degeneracy) in monolayer graphene.5,6 Such an extraordinary quantization
phenomenon is related to the quantum anomaly of the n = 0 LL coming from the Dirac point.1
Currently, scientists are in search of novel materials yielding unusual physical phenomena and
having potential for applications. By generating graphene-based defect systems, such as
substituting impurities or guest atoms in a hexagonal carbon lattice, ones could effectively
manipulate the fundamental properties inherent to graphene. Among various pertinent guest
atoms, Si,23 B,24,25 and N26,27 have been successfully substituted for the carbon host atoms by
chemical vapor deposition (CVD) or arc discharge experiments. The doping effect eliminates the
equivalence of the two sublattices, causing dramatic changes in physical properties, including the
energy gap opening and the deviation from the original Dirac cone.
Previous studies have proved the emergence of different ionization potentials and non-
uniform hopping integrals.28,29 Specifically for Si-, B-, and N-doped graphene, the π bonding
extending on a hexagonal lattice is significantly distorted. As a consequence, the low-lying
energy band is controlled by both the dopants and C atoms simultaneously. The magnetically
quantized LLs of the dopant- and concentration-created unusual band structures have been
theoretically predicted to be feature-rich and unique.30 These facts make it promising for the
QHE of such special LL energy spectra to yield abundantly rich results.
The rest of our paper is outlined as follows. In Sec. II, we employ the tight-binding
method to solve for the eigenvalues and eigenfunctions of the magnetic Hamiltonian, and then
use these results to calculate the Hall conductivity by means of the linear Kubo formula. This
ISBN: 978-81-948129-1-3 Page 309
procedure enables us to identify allowed transition channels through the aforementioned

magneto-electronic selection rules, leading to the understanding of Fermi-energy- and magnetic-
field-diversified quantum conductivity. Fundamental properties of doped graphene are greatly
enriched by various types of dopants and their concentrations. In Sec. III, we investigate the
QHE of several dopants of Si, B, and N with 2% and 12.5% of doped atoms. Our results
demonstrate the robust correlation between the unusual QHE and the dopantand concentration-
induced rich LLs in the doped graphene systems. Our theoretical predictions provide essential
information for future experimental verification of the QHE in graphene-based defect materials.
Section IV is devoted to our concluding remarks.
II. METHOD
The crystal structures of doped graphene for various concentrations are presented in Figs.
1(a) and 1(b). For a 1:2n 2 concentration doping (n is the cell multiplicity), the super cell could
be obtained by expanding the original unit cell of pristine graphene n times along the lattice
vectors ~a1 and ~a2, that is, (n ~a1, n ~a2). In this work, we choose a rectangular super cell as
marked by the red lines for convenience in considering the effect of magnetic field. A single
supercell comprises 2n 2 sublattices of two types, Ai and Bi , in which i indicates the lattice site.
Both C-2pz orbital and the guest orbitals of B − 2pz, Si − (2pz, 3pz), and N − 2pz play 3 the key
roles in governing the crucial characteristics at low-energy range. The Hamiltonian matrix
includes the non-uniform bond lengths, site energies and nearest-neighbor hopping integrals,
which could be written as follows:30
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In this notation, f1,2,3 is defined in terms of the wave vector ~k and the vectors
connecting the nearest-neighbor lattice sites R~ 1,2,3 through e i~k.R~ 1,2,3 . m(k) = k + n − 2
mod n is modulo function, and j, l are the integers (j, l = 1, 2, 3, ..., n).
The C-C and B/Si/N-C bond lengths have been verified as being slightly different due to
doping, especially the buckling effect of the guest atoms in graphene.28,29 For Si dopant, the
guest atoms are located at a distance of dSi above graphene sheet, implying the significant
adjustment of the π bonding on a hexagonal lattice. The consequential non-uniform
nearestneighbor hopping integrals and site energies associated with the major 2pz orbitals of the
C host atoms and the minor 2pz/3pz orbitals of the guest atoms lead to extraordinary electronic
properties. The parameters are appropriately fitted, as shown in Table I, so that the energy bands
calculated by the tight-binding and first-principles methods are consistent.
The effect of a uniform perpendicular magnetic field on the system could be treated with
the use of the vector potential-created Peierls phase of ∆Gmm0 = 2π φ0 R Rm Rm0 A(r) · dr (φ0
= hc/e the flux quantum) in the nearest-neighbor hopping integral.30 Such a phase is position-
dependent so that the unit cell is extended to become along the armchair direction. Consequently,
the magnetic Hamiltonian is a large matrix whose periods strongly depend on the density of
guest atoms and vector potential. This Hamiltonian matrix is efficiently
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FIG. 1: (Color online) Crystal structures of doped graphene for chosen doping concentrations of
(a) 12.5% and (b) 2% dopants. The green and orange spheres denote, respectively, the C and
Si/B/N atoms.solved using the diagonalization method to obtain the LL energy spectrum and
sub-envelope functions.31 Interestingly, the amplitudes of sub-envelope functions on the distinct
sublattices in an enlarged unit cell could be regarded as their spatial distributions, and they are
used, therefore, to analyze the main features of the LL wave functions.
In this notation, |αi and Eα are, respectively, the LL state with energy, fα,β the Fermi-
Dirac distribution functions, S is the area of the supercell, and Γ0 a broadening parameter.
Furthermore, the velocity operators ˙ux,y directly determine the allowable inter-LL transitions;
they are evaluated from the gradient approximation .33 The defect effect in doped graphene
might induce the unconventional quantum conductivities.
III. RESULTS AND DISCUSSION
The effect of doping with chosen dopants and concentrations remarkably diversifies the
electronic properties of graphene. The substitution of guest atoms opens a direct band gap in
monolayer graphene since it breaks the lattice symmetry, as illustrated in Figs. 2(a) through 2(f).
The magnitude of the energy gap (Eg) is strongly determined by the dopants and doping
concentration, as demonstrated in Table II for pristine graphene and Si-, B-, and N-doped
systems with 2% and 12.5% guest atoms. Additionally, the significant influence of doping on the
Fermi energy is worthy of consideration (Table III). For pristine graphene, the Fermi energy is
equal to zero (Fig. S1 of the Supplemental Materials) and this remains the same in the presence
of Si dopant. On the contrary, the Fermi energy levels are downshifted to the valence band or
upshifted to the conduction band for B- and N-doped systems, respectively. These phenomena
could be explained by the fact that a B atom acts as an p-dopant while an N atom is an n-dopant
since a B/N atom has one less/more electron than a C atom. The 3D plots for energy bands with
separation of occupied and unoccupied states are shown in Fig. S2 of the Supplemental
Materials. Our numerical results are in
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FIG. 2: (Color online) Energy bands of doped graphene for Si, B, and N guest atoms with (a)-(c)
2% and (d)-(f) 12.5% concentrations, respectively.
Agreement with previous theoretical calculations by the first-principles method.34–36 The
prediction of the dopant- and concentration-dependent band structures in doped graphene should
be useful for technological applications.
In the presence of a uniform perpendicular magnetic field, the quantized LLs show unusual
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Characteristics due to the doping effect. Because of the opening of a band gap, the conduction
and valence LL energy spectra are clearly separated, as shown in Figs. 3(a) and 3(d) for the
magnetic-field-dependent energy spectra of Si-doped graphene. The separation is invariant with
variation of the field strength so that it has no influence on the inter-LL transition between the
valence and conduction bands. Each LL is four-fold degenerate, except for the lowest conduction
and highest valence states in the vicinity of zero energy whose degeneracy is half smaller. For
Si-doped graphene, the Fermi level is located at zero energy. Therefore, the former and the latter
are unoccupied and occupied, respectively. On the other hand, they are either unoccupied for B
or occupied for N dopants due to the shift of Fermi level. These two doubly degenerate LLs are
split from the original zero-energy LL (n = 0) of pristine graphene due to the substitution of
guest atoms. The Bz-dependence of LLs near the Fermi levels in B- and N-doped systems are
worthy of detailed consideration. There, the density of LL states is very high, as clearly
illustrated in Figs. 3(b) through 3(c) for 2% and 3(e) through 3(f) for 12.5% concentrations of B
and N guest atoms. Especially for sufficiently high dopant density of 12.5%, the LLs show slight
splitting behavior at higher and deeper energy ranges, altering the state degeneracy degree of
freedom. These phenomena play a key role in the unique properties of quantum Hall
conductivity, which we discuss below. The contribution of guest and host atoms to the Hall
conductivity through the velocity matrix elements in Eq. (3) could be understood based on the
amplitude of LL wave functions on the sublattices, as demonstrated in Figs. 4(I) through 4(III).
Our numerical calculations
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FIG. 3: (Color online) The Bz-dependent LL energies of doped graphene with Si, B, and N
dopants for (a)-(c) 2% and (d)-(f) 12.5%.
show that the A sublattice which consists of the guest atoms obviously dominates the conduction
bands of (Si, B)-doped systems and the valence band of N-doped graphene, and vice versa for
the B sublattice. Especially for Si dopant with the presence of 3pz orbital, its spatial distribution
is even stronger than that of the C atoms. The quantum number, which
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FIG. 4: (Color online) The LL energies and wavefunctions of the first few levels for (I) Si, (II) B,
and (III) N guest atoms with 12.5% concentrations for Bz = 20 T. The blue and red bars
represent LLs localized at 1/6 and 2/6 centers, respectively.
It is critical in understanding the selection rule of inter-LL transition, is determined based
on the LL spatial distribution. Landau wave functions are only localized at some specific areas
within the field-enlarged unit cell, particularly at 1/6, 2/6, 4/6 and 5/6 centers for a (kx = 0, ky =
0) state. LLs localized at 1/6 and 4/6 (2/6 and 5/6) are equivalent in the main characteristics and
they are classified as n c,v 1 (n c,v 2 ) group. This is true for all the doped systems, regardless of
the dopants and doping concentration. The doubly-degenerate conduction LL belongs to n c 1
group for Si and B dopants but n c 2 for N, while the opposite is true for the valence one.
The quantum Hall effect of doped graphene presents unusual and unconventional properties that
may be manipulated by the degree to which the sample is doped. The Fermi energydependent
conductivity quantization in an external magnetic field presents step structure, as illustrated in
Fig. 5 for the resistivity Rxy = 1/σxy of three different guest atoms with 2% and 12.5%
concentrations. The plots of σxy could be found in Fig. S3 of the SupplementalMaterials. During
the variation of the Fermi energy (EF ), a plateau is formed whenever one LL becomes occupied.
The inter-LL transition between the occupied and unoccupied levels needs to satisfy a specific
ISBN: 978-81-948129-1-3 Page 316

selection rule where the oscillation mode of the wave functions on the sublattices must be
equivalent for the initial and final states. We observe that such a rule is ∆n = ± 1 & 0 for 2% and
∆n = ± 1 for 12.5% doped graphene.
FIG. 5: (Color online) The EF -dependent resistivity of Si-, B-, and N-doped graphene for Bz =
20 T. The solid and dashed curves represent the dopant densities of 2% and 12.5%, respectively.
The inset shows the quantum Hall conductivity of 12.5% B-doped graphene.
The conductivity is quantized following a special sequence of σxy = {0; 2(2m + 1)e 2/h} (m is
an integer) for all dopants. It should be noticed that, the EF on the x-axis in Fig. 5 represents the
shift of energy from the original Fermi level. The band gap created by the doping is reflected in
the discontinuity of Rxy near the zero energy, as shown for Si guest atom by the solid and
dashed red curves. This is associated with the emergence of zeroconductivity, clearly
demonstrated in Figs. S3(a) and S3(d) of the Supplemental Materials. As a result, there exist two
half-integer steps of 2e 2/h height near zero energy, in addition to the conventional steps of 4e
2/h. This is in contrast with that for pristine graphene where the two half-integer steps at zero
energy are not separated.5 The width of the plateau at zero energy, which directly reflects the
size of the band gap, strongly depends on the dopants and doping concentrations.
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The quantum Hall conductivities in the vicinity of the Fermi levels in the presence of B and N
dopants deserve careful scrutiny since they could be identified in magneto-transport
measurements.
For sufficiently small doping concentration, e.g., 2% as illustrated by the solid blue and
green curves in Fig. 5 (also in Figs. S3(b) and S3(c)), the quantized sequence of quantum Hall
conductivity remains unchanged even at higher or lower energy ranges. The doping effect might
not be strong enough to remarkably alter the essential physical properties of the system.
However by increasing the density of dopants, the ensuing split of LLs near EF (B) and EF (N)
gives rise to the significant changes in quantum Hall effect. There, the wider and smaller
conductivity plateaus are interspersed in the spectra, referring to the inset plot for 12.5% B-
doped graphene (also in Figs. S3(e) and S3(f)), in which the latter is related to the splitting of
degenerate LLs. Accordingly, each step is reduced by half in height, obeying a new quantized
sequence of σxy = 2me2/h.
Our theoretical predictions for doped graphene of various dopants should provide useful
information for further magnetic transport measurements, as done for monolayer and few-layer
graphene.5,6,8–11 Specifically, we observed the half-integer QHE as predicted to exist in
monolayer graphene by different theory groups,1–3 situated on thin graphite films,4 and verified
by experimental measurements.5,6 Such a special quantization in graphene is attributed to the
quantum anomaly of the n = 0 LL corresponding to the Dirac point.1 The results obtained in this
work by the well-developed theoretical framework are expected to be confirmed experimentally.
IV. CONCLUDING REMARKS
In conclusion, the generalized tight-binding model and the Kubo formula are employed to
explore the interesting QHE in doped graphene for Si, B, and N guest atoms with various
concentrations. The theoretical framework takes into consideration substantial factorsof the
doping system, including the non-uniform bond lengths, site energies, hopping integrals, and
external field. This method could be further developed to investigate many other condense-
matter materials. The doped graphene is an emergent 2D binary compound material, which
presents diverse electronic and transport properties under magnetic quantization. The doping
effect can open a band gap of different sizes, leading to remarkable changes in the main features
of LLs and thus the magneto-transport properties. The Fermienergy-dependent QHE exhibits
both the integer and half-integer conductivities. The step structure could be controlled by
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changing the dopants or doping concentration, in which the latter has more significant influence.
The theoretical predictions should provide useful information and could be examined by
magneto-transport measurements.
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43. S. S. Varghese, S. Swaminathan, K. K. Singh, and V. Mittal, Electronics 5, 91 (2016).
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A DETAILED REVIEW ON WOUND DRESSING APPLICATION OF CHITOSAN

BASED BIOMATERIALS
Pradeep Kumar Panda1, Pranjyan Dash2 and Zoyeb Mohamed Zia3
1
Department of Chemical and Materials Engineering, Chang Gung University, Taoyuan, 33302,
Taiwan, Email: rkpanda277@gmail.com
2
Department of Chemical and Materials Engineering, Chang Gung University, Taoyuan, 33302,
Taiwan Email: pranjyandash@gmail.com
3
PG and Research Department of Zoology, The New College Chennai,
Email: zoyebmdzia@gmail.com
ABSTRACT:
Fighting bacterial resistance is one of the concerns in modern days, as antibiotics remain
the main resource of bacterial control. Data shows that for every antibiotic developed, there is a
microorganism that becomes resistant to it. Natural polymers, as the source of antibacterial
agents, offer a new way to fight bacterial infection. The advantage over conventional synthetic
antibiotics is that natural antimicrobial agents are biocompatible, non-toxic, and inexpensive.
Chitosan is one of the natural polymers that represent a very promising source for the
development of antimicrobial agents. In addition, chitosan is biodegradable, non-toxic, and most
importantly, promotes wound healing, features that make it suitable as a starting material for
wound dressings. This paper reviews the antimicrobial properties of chitosan and describes the
mechanisms of action toward microbial cells as well as the interactions with mammalian cells in
terms of wound healing process. Finally, the applications of chitosan as a wound-dressing
material are discussed along with the current status of chitosan-based wound dressings existing
on the market.
KEYWORDS: chitosan; antimicrobial properties; mechanism of action; wound dressing; wound
healing
1. INTRODUCTION
According to the Merriam-Webster Medical Dictionary, an antimicrobial is an agent that
either prevents and stops the growth of pathogen microorganisms or even kills them [1].
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Antimicrobial agents have been used since at least 2000 years ago, when the Ancient Egyptians
and Greeks used different plant extracts and mold to treat infections [2]. Since the well-known
discovery of penicillin by Alexander Fleming in 1928 through to modern days, antibiotics have
been among the most commonly used drugs for treating pathogen infections [3]. Since the
number of new classes of antibiotics has decreased and no new classes have been discovered
since daptomycin and linezolid in the 1980s, optimizations of already-existing molecules or
combinations of different compounds are used for new commercialized products [4]. Figure 1
presents a schematic representation of the mechanisms of action traditionally targeted by
antibiotics: (1) disruption of the cell membrane, (2) inhibition of nucleic acid synthesis
(DNA/RNA), (3) inhibition of protein synthesis by acting on ribosomes, (4) regulation of
enzymes involved in cell wall biosynthesis, and (5) inhibition of folic acid synthesis in the
cytoplasm. A modern approach in order to combat antimicrobial resistance would be finding
newer targets or using alternative antimicrobials, either natural or synthetic compounds [5,6].
The need for developing new generations of antimicrobial agents is due to the existence
of drug-resistant pathogens and the emerging problem of microbial infections. Infections with
antibiotic-resistant pathogens are not only difficult to treat but also involve prolonged debility for
patients and expensive healthcare [7]. Studies show that more than 70% of chronic wounds are
exposed to microbial biofilm infections [8]. A microbial biofilm is described as a well-
structured, three-dimensional microbial aggregate that surrounds itself with a self-produced
exopolymer matrix containing polysaccharides, proteins, and nucleic acids [9–11]. The
resistance of biofilm to antibiotic treatments may be explained by various molecular mechanisms
[12]. These include, among others, (1) limited diffusion of drugs through the exopolysaccharide
matrix [13], (2) the presence of persister cells, which will allow the repopulation of biofilms
following antibiotic treatment [14], and (3) possibility of exchanging antibiotic-resistant genes
between organisms within the biofilm [12]. Biofilms are by far less susceptible to antibiotics,
compared to planktonic cells, and quorum-sensing molecules regulate biofilm formation and
expression of specific properties.
Also called autoinducers (AIs), quorum-sensing molecules, which regulate most
virulence factors, have been identified in various bacterial species. Staphylococcus aureus and
Pseudomonas aeruginosa, two species intensively studied regarding quorum-sensing molecules,
are also commonly associated with chronic wound infections [10]. Treating wound infections is
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more difficult when a biofilm is present. The extracellular polymeric matrix protects the biofilm
from inflammatory processes, which are crucial steps in wound healing [15]. The phagocytosis
process and complement cascade activation are blocked and the exopolymer matrix represents
itself a physical barrier against antimicrobial drug penetration into the wound. Leucocyte
penetration and movement are also limited through biofilms; therefore, their ability to produce
ROS (reactive oxygen species) is impaired [10].
Figure-1
Besides commonly used antimicrobials such as antibacterials, antifungals, antivirals, and
antiparasitics, chemical and natural compounds are some other types of antimicrobial agents that
are of great interest not only to the scientific community but also to the public in general, given
the fact that antimicrobial resistance is a worldwide threat. In 2018, the European Centre for
Disease Prevention and Control (ECDC) stated that nearly 33,000 die every year as a
consequence of an infection with antibiotic-resistant bacteria [16]. Therefore, natural and
synthetic molecules are an important source for developing new antimicrobials. Antimicrobial
polymers are considered the new generation of antimicrobials. Such polymers include, among
others, antimicrobial peptides, halogen-containing polymers, phosphor and sulfur derivatives,
metal nanoparticles, dendrimers [17],heparin, poly-ε-lysine, gramicidin A, quaternary
ammonium polyethyleneimine, and guanylated polymethacrylate, but the most promising is
chitosan [18].
Chitosan, a natural cationic polysaccharide consisting of (1→4)-2-amino-2-deoxy-β-d-
glucan, is the partially- to fully-deacetylated form of chitin [19]. For many years, there has been
great interest in the use of chitin since this biopolymer is the second most abundant after
cellulose [20]. There are various sources of chitin: crustacean shells, fungi and algae cell walls,
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insect exoskeletons, and mollusk radulae [21–24]. Commercial chitin, (1→4)-2-(acetylamino)-2-

deoxy-β-d-glucan, is obtained, following demineralization and deproteinization processes,
mainly from crustaceans shells [25,26], which are byproducts of the seafood-processing industry.
The seafood-processing industry creates a large quantity of waste every year. About 75% of the
total body mass of crustaceans ends up as byproducts (head, tail, backbone, and shell) [27]. Due
to a lack of better management of waste, most often, seafood raw materials are discarded back
into the sea, burned, or just left out to deteriorate, causing environmental problems [28,29].
Therefore, the extraction of chitin and other valuable substances from crustacean waste is an
alternative that can reduce the waste and is an important step in producing useful compounds that
have important biological features and applications in various fields. According to The State of
World Fisheries and Aquaculture 2018 report by the FAO (Food and Agriculture Organization of
the United Nations), global farmed food fish production included approximately 7 million tons of
crustaceans [30]. After the edible part of crustaceans has been removed, the remaining material
represents an important source of chitin as well as proteins, lipids, pigments, and other molecules
[28,31–33].
Figure-2
Unlike chitin, chitosan is soluble in acidic aqueous solution due to its acetylation degree.
The acetylation degree of chitosan can be expressed either as a percentage of acetylation (DA%)
or as the molar fraction of N-acetylated units (DA). Therefore, when the DA is lower than 50%
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(0.5), dissolution is possible due to the protonation of the amino group at the C-2 position of
glucosamine units [34].
The physical-chemical characteristics of chitosan are important in order to understand its
functional properties. For example, the degree of acetylation, which refers to the distribution of
amino groups along the polymer chain, determines one of the most important features of
chitosan, its polycationic nature in acidic media, resulting from the ionization of amino groups.
Thus, functional properties of chitosan, such as solubility, swelling ratio, bioactivity, and
biodegradation, are influenced by the degree of acetylation. The molecular weight together with
the deacetylation degree is the most important physical-chemical characteristics of both chitin
and chitosan. By manipulating the molecular weight of chitosan, functional properties can be
controlled; for example, viscosity can be reduced and water solubility can be enhanced [35].
Chitosan is also an important tool in research fields due to its significant biological
properties such as biocompatibility and biodegradability [36–40], low toxicity [41–45], and
antimicrobial [46–52], antioxidant [49,53–56], and anti-cancer [57–62] properties. Due to all of
the valuable features mentioned above, chitosan is widely used in medicine and pharmaceutical
fields, the food and agriculture industry, as well as cosmetics and waste treatment [63–66].
In this review article, we will further discuss chitosan antimicrobial properties and describe its
mechanism of action against microbial cells, as well as the interactions with mammalian cells.
2. CHITOSAN MECHANISM OF ACTION AGAINST PATHOGEN
MICROORGANISMS
Chitosan is one of the most promising natural polymers known to possess antimicrobial
properties [67,68]. Interest in the antimicrobial properties of chitosan began many years ago, and
some of the first studies that described antimicrobial features of this biopolymer date back to the
1980s and 1990s [69,70]. The main conclusion highlighted by these studies was that chitosan has
a bacteriostatic and bactericidal effect, often without making any distinction between these two
terms. More recent studies tend to focus more on the bacteriostatic effect of chitosan, although
the exact mechanism of action against pathogens is not well known [71,72]. According to the
literature, there are a few main generally accepted mechanisms of action of chitosan against
microorganisms, which are described below.
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2.1. DISRUPTING THE CELL MEMBRANE/CELL WALL

Probably the most discussed mechanism refers to the alteration of cell permeability and
lysis of the cell membrane. This effect is presumably caused by electrostatic interactions between
the positively charged chitosan molecule and negatively charged cell membrane [73–84]. The
antimicrobial activity of chitosan is based on the presence of the amino groups of the polymer
chain. These amino groups can be protonated, giving chitosan a positive charge [78]. Therefore,
chitosan becomes soluble in aqueous acidic solutions when the pH value is lower than its pKa
value of 6.3, in which case the –NH2 groups are converted to a soluble protonated form −NH+ 3
[85,86]. The cell wall of Gram-positive bacteria is composed mainly of a thick peptidoglycan
layer containing teichoic acids, which give a negative charge to the bacterial surface, while the
Gram-negative bacteria cell wall is strongly negatively charged due to lipopolysaccharides
contained in the outer membrane layer. In addition, a similar situation occurs in the case of the
fungal membrane and viral envelope, which contain some negatively charged compounds [87].
Fu et al. showed that teichoic acids contained in the cell wall of Staphylococcus aureus interact
with chitosan derivatives, thus inhibiting the growth of the bacteria, in some cases up to
approximately 100% [88]. Li et al. demonstrated that the cell membrane of Escherichia coli was
damaged by chitosan.
They demonstrated extracellular leakage by measuring absorption at 260 nm and 280 nm,
estimating, in this way, the amount of DNA and proteins released from the cytoplasm. In
addition, electron micrographs obtained using transmission electron microscopy (TEM) revealed
clear differences between E. coli cells treated with chitosan and those used as a control.
Chitosan-treated bacterial cells showed a seriously altered outer cell membrane with
cytosolic components coagulated after treatment for 24 h [89]. Vallapa et al. demonstrated that
the introduction of additional positive charges to chitosan via heterogeneous quaternization
increased the antibacterial activity against both Gram-positive and Gram-negative bacteria,
compared with native chitosan [90]. Chitosan may also exhibit antimicrobial activity against
fungi. Here, the microorganisms can be divided into two categories: chitosan-sensitive fungi and
chitosan-resistant fungi. In this case, the mechanism of action is slightly different from that of
bacteria. First, chitosan molecules interact with negatively charged phospholipids, thus affecting
the cell membrane and causing the leakage of intracellular material. However, in the case of
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chitosan-resistant fungi, this polymer seems to be unable to penetrate the cell membrane, and it
remains outside the cell surface [91,92].
The fluidity difference of the cell membrane is presumably the reason why chitosan is
unable to permeate the cell membrane. The plasma membrane of chitosan-sensitive fungi has
more polyunsaturated fatty acids than chitosan-resistant fungi, which indicates that cell
membrane disruption by chitosan may depend on the fluidity of membranes [78]. Palma-
Guerrero et al. showed that the phospholipid fatty acid composition of the cell membrane is
related to chitosan antimicrobial activity. To demonstrate this, they tested the antimicrobial
activity of chitosan against a fatty acid desaturase mutant strain of Neurospora crassa, which
exhibited increased resistance to chitosan compared to the wild type [92].
2.2. INTERACTION WITH MICROBIAL DNA
Another mechanism consists of the interaction of chitosan hydrolysis products with
microbial DNA, thus affecting protein synthesis by inhibiting mRNA [78,93,94]. The ability to
penetrate a cell’s membrane and to enter in the cell was observed in the cases of low molecular
weight chitosans [72,95–97]. Xing et al. investigated the binding of oleoyl-chitosan nanoparticles
(OCNPs) to DNA/RNA by examining the effect of OCNPs on the electrophoretic mobility of
nucleic acids. The results showed that once the concentration of OCNPs increased, the
interactions between bacterial genomes multiplied, and by the time OCNP concentration reached
1000 mg/L, the migration of E. coli DNA and RNA was completely inhibited. It is assumed that
the negatively charged phosphate groups in the chain of nucleic acids interact with the positively
charged amino groups in OCNPs, thus affectingpathogen activity [83]. Galván Márquez et al.
suggested that chitosan inhibits protein biosynthesis by testing β-galactosidase expression assays.
Their experiments showed that β-galactosidase activity in yeast cells was reduced by 13% when
chitosan concentration was 1.25 mg/mL [95].
2.3. CHELATION OF NUTRIENTS BY CHITOSAN
The high chelating capacity of chitosan towards various metal ions (Ni2+, Zn2+, Co2+,
Fe2+, Cu2+) when the pH value is higher than its pKa value could explain the inhibitory effect
of chitosan towards microbial growth [19,71,72,98–100]. Both Gram-positive and Gram-
negative bacteria cell wall components attract divalent metal cations that are beneficial to the
microbial cell. The phosphate groups of teichoic acids, contained in the peptidoglycan layer of
Gram-positive bacteria, attract especially Mg2+ and Ca2+ cations, which help to maintain
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enzymatic functions and the integrity of membranes. Lipopolysaccharides, components of Gram-

negative bacteria cell surfaces, give the bacterial cell membrane a negative charge and also have
a strong affinity for divalent cations [78]. The lack of these divalent cations makes bacteria more
sensitive to chemicals or certain antibacterial agents. Chelating agents, such as
ethylenediaminetetraacetic acid (EDTA) or some weak acids, can remove these cations, making
the cell wall a more permeable and less effective barrier [101]. It is known that chitosan has
chelating properties, so it can chelate metal ions and essential nutrients that are important for the
growth of the microbial cell [102–104]. At a low pH (below 6.0), the protonated amino groups in
the chitosan polymer chain compete with the divalent cations for the phosphate groups found in
the lipopolysaccharide or teichoic acid structure. Mansilla et al. showed that the addition of
Mg2+ and Ca2+ to the culture medium increases the positive charge of the membrane and
weakens the antimicrobial action of chitosan, proving once again that the bacteriostatic action
depends mainly on electrostatic interactions [96].
2.4. FORMATION OF A DENSE POLYMER FILM ON THE CELL SURFACE
High molecular weight chitosan may deposit and form a dense polymer film on the
surface of the cell, preventing nutrient and oxygen uptake, which then inhibits the growth of
aerobic bacteria [93,105–107]. Champer et al. observed bacterial cells covered with chitosan
aggregates using an electron and fluorescence microscope [108]. Due to chitosan deposition, cell
walls appeared thicker and covered by an additional layer of vesicular structures. The thickness
of the outer membrane prevents the nutrients from entering the cell and also prevents the
excretion of metabolic products [78].
3. CHITOSAN INTERACTION WITH PROKARYOTIC CELLS
Often, bacteria need to survive even in the most hostile environments; therefore, they developed
a specialized cell envelope that is selectively permeable and protects the cell.
Electrostatic interaction between positively charged chitosan molecules and negatively
charged lipopolysaccharides (Gramnegative bacteria) and teichoic acids (Gram-positive bacteria)
may lead to the blocking of intra/extracellular exchanges or even cell wall disruption and, finally,
leakage of cytoplasmic content.
3.1. INTERACTION WITH GRAM-POSITIVE BACTERIA
The difference between Gram-positive and Gram-negative bacteria is in their cell wall
structure. When referring to Gram-positive bacteria, the main feature is the presence of teichoic
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acids, which can interact with chitosan molecules. Teichoic acids are anionic glycopolymers
linked to the peptidoglycan layer that, among other functions, play an important role in
pathogenesis, modulating susceptibility to cationic molecules. There are two types of teichoic
acids: wall teichoic acids and lipoteichoic acid, which are anchored in the bacterial cell
membrane. The resistance of Gram-positive bacteria to antimicrobial agents is through teichoic
acids and their attached substituents that regulate the negative charge of the bacterial cell, which
also prevents the binding of extracellular molecules [109]. This may also be the actual
mechanism through which chitosan interacts with the Grampositive bacterial cell wall. Being
positively charged, chitosan can interact with negatively charged teichoic acids, altering cell wall
rigidity, disrupting the cell membrane, and eventually entering the cell. Moreover, the presence
of D-alanine esters attached to teichoic acids contributes to the bacterial surface charge, and the
reduction of D-alanyl content in the cell wall confers susceptibility to glycopeptide antibiotics
and some cationic antimicrobial peptides [110]. Studies revealed that the absence of D-alanyl
esters can increase the negative charge of the cell surface and facilitate the attraction of cationic
molecules, thus increasing the sensitivity of the cell to such antimicrobials.
3.2. INTERACTION WITH GRAM-NEGATIVE BACTERIA

Gram-negative bacteria have a more complex cell surface structure consisting of an outer
lipid bilayer structure, the peptidoglycan cell wall, and the plasmatic membrane. The outer lipid
bilayer is a distinct feature of Gram-negative bacteria, and its structure contains phospholipids in
the inner layer and a single type of glycolipids, known as lipopolysaccharides (LPSs), in the
outer layer. The outer layer is a selective barrier with two important features: (1) the selectivity,
provided by the porins, which are small protein channels that diffuse certain hydrophilic
molecules, and (2) the high negative charge, which is given by LPS molecules.
The structure of LPS consists of the lipid A and the inner core, which are the negatively
charged groups responsible for the interactions with cationic polymers. Lipid A can also be one
possible target for neutralizing the endotoxic effect of LPS [112,113]. Davidova et al.`s study
revealed that the interactions occur amongst LPS and the amino groups of chitosan. This was
demonstrated using the anionic dye tropaeolin. Experiments showed that at pH value between 4
and 7, where the amino groups of chitosan are ionized, the LPS endotoxins displaced the
negatively charged dye molecules from the complex they formed with this cationic polymer
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[110]. Since porins are known to be responsible for the uptake of nutrients [114], it is possible
that chitosan molecules can block the exchange of nutrients by forming a polymer layer on the
surface of the cell, thus leading to bacterial cell death.
4. CHITOSAN INTERACTION WITH EUKARYOTIC CELLS
4.1. INTERACTION WITH FUNGAL CELLS
As many studies show, chitosan can inhibit bacterial cell growth and even kill them, but
regarding yeast cells, only high concentrations of chitosan can have an antifungal effect and
inhibit yeast cells growth [71,87,115,116]. Since chitosan’s cationic nature is the result of the
presence of reactive amino groups in its structure, if these groups are blocked, antimycotic
activity will be severely reduced [117]. The fungal cell wall is a unique structure, and its
components are glycoproteins and polysaccharides, mainly glucan and chitin. These structures
are strongly cross-linked to form a resistant assembly that will protect the fungal cell from
environmental stress but at the same time will allow the fungal cell to interact with its
environment
Each component of the cell wall is responsible for ensuring cell integrity. For example, if
chitin synthesis is perturbed, the fungal cell wall becomes osmotically unstable and deformed.
The synthesis of β-1, 3-glucan is necessary for the normal development of fungal cell walls, and
the glycoproteins are responsible for maintaining cell shape, intracellular signal transmission,
synthesizing other cell wall components, and facilitating molecules absorption, but at the same
time protecting the cell against foreign substances [118].
The organization of glycan and proteins at the surface of the cell is unique for each fungal
species. The phosphate groups, located in the carbohydrate side chains of highly glycosylated
cell wall proteins, give the negative charge at the surface of the fungal cell [119]. Studies have
demonstrated the presence of sialic acid on the surface of Candida albicans cell walls [118,120].
Anionic groups at the surface of the fungal cell give a high negative charge that could facilitate
electrostatic interactions with different molecules and the host cell [121]. As mannoproteins are
the major components at the surface of the cell wall and the sialic acids are considered terminal
residue in the carbohydrate side chain of mannoproteins [119], electrostatic interactions could
occur between chitosan and the fungal cell surface. Soares et al. demonstrated that sialic acids
contributed to the electrical negative charge of the cell surface by facilitating the adherence of
fungal cells to a cationic substrate (poly-l-lysine) [120].
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The mechanism by which chitosan inhibits fungal growth is not fully understood yet, but some
proposed theories are described below and represented in Figure 3:
(a) Chitosan is a cationic polymer that will interact with the negatively charged cell surface (due
to the presence of sialic acid) and can destabilize the electrical charge of the cell wall;
(b) Once the negative charge is decreased, the concentration of the cations at the surface is also
affected (mainly K+), and the difference between internal and external concentration leads to the
efflux of cations;
(c) This efflux of cations will further cause the hyperpolarization of the plasma membrane,
therefore causing an increased uptake of Ca2+ and the loss of negatively charged molecules
(nucleotides, phosphates, substrates for enzyme reaction), affecting the metabolism of the cell
and altering important metabolic pathways [122,123].
Figure 3.Schematic representation of chitosan mode of action on a Candida albicans cell.

The fungal cell surface is negatively charged due to the carbohydrate side chains of
mannoproteins, mainly sialic acids. Cationic chitosan molecules can cause ionic interactions with
anionic groups and destabilize the cell wall. Other mechanisms of action proposed in the
literature are metal chelation, enzyme denaturation, and chitosan interaction with phosphate
groups of nucleic acids, all causing growth inhibition or even microbial death.
Moreover, chitosan may form a dense film on the cell surface, which may interfere with
fungal growth and activate several defense mechanisms such as chitinase accumulation, callus
synthesis, and proteinase inhibitor synthesis and lignification [43]. Another hypothesis, by which
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yeast cells may recover after long exposure to chitosan or even gain resistance, is the existence
of chitinases that may degrade deacetylated chitosan [124,125]. In their study, Oliveira Junior et
al. reported that chitosan caused morphological changes and abnormal shapes of fungal mycelia.
They tested the effect of chitosan with different molar fractions of acetyl groups (FA 0.16 and
0.18) and degrees of polymerization (DP 1089 and 1242), and both types of chitosan had a direct
influence on the three fungal species studied (Alternaria alternate—500 µg/mL, Botrytis
cinerea—1000 µg/mL, Penicillium expansum—500 µg/mL). Besides causing a delay in their
growth, the micrographs also showed mycelial aggregations, excessive branching, hyphae size
reduction, and swelling of the fungal cell wall [109]. Yien et al. concluded that C. albicans
proved to be more susceptible to chitosan nanoparticles compared with other species [122]. One
argument could be the presence of negatively charged sialic acids in the cell wall [120].
4.2. INTERACTIONS WITH MAMMALIAN CELLS IN WOUND HEALING
PROCESSES
Despite the advanced development of antiseptics, infections of wounds remain a major
issue. The emergence of multidrug-resistant pathogens leads to a constant need for more efficient
topical antimicrobial products. The main drawback is that most of them are highly cytotoxic,
affecting the tissue healing process [126].
Apart from antimicrobial and antifungal properties, chitosan is innate, biocompatible, and
non-toxic to living cells and tissue. Chitosan biocompatibility has been tested in vitro using
different types of cells like fibroblasts, keratinocytes, and hepatocytes, as well as myocardial and
endothelial cells [127].
Wounds heal naturally, but there are persons that suffer wound healing disorders, such as
diabetic patients who may develop chronic, non-healing wounds that subject the patients to long-
term distress and discomfort [113]. In such cases, a long treatment time is required, which will
also increase the costs associated with advanced medical care. Therefore, the focus has been on
natural therapeutic substances that can accelerate the wound healing process and at the same time
be easily accessible to the general population, and chitosan seems to meet all these conditions.
Chitosan accelerates the wound healing process by stimulating inflammatory cells, macrophages,
and fibroblasts, hence boosting the inflammatory phase. In this way, the inflammatory phase is
reduced, and the proliferative phase starts sooner in the wound healing process [128].
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Caetano et al. investigated the healing properties of a chitosan–alginate membrane on a

cutaneous wound in rats and observed the following: the wound was not infected, fibroplasia was
significantly increased, the fibroblasts were arranged better in the newly formed tissue, and the
quality of scar tissue was improved. Moreover, on the 7th day of treatment with the chitosan–
alginate membrane, the inflammatory infiltrate was significantly reduced, followed by a decrease
of neutrophils and CD4+ lymphocytes that might indicate a better regulation of the inflammatory
stimulus by the chitosan–alginate membrane. Likewise, the chitosan–alginate complex
stimulated the migration of CD11B+ macrophages. These cells are very important as they
continue the work of neutrophils, acting as growth factor reservoirs and releasing several
enzymes that digest the remaining extracellular content, facilitating the transition to the second
phase of the healing process [112]. Chitosan may also have the ability to regulate granulation
tissue formation and angiogenesis, assuring the correct deposition of collagen fibers and further
enhancing the correct repair of injured dermal tissue [129]. When referring to the wound healing
properties of chitosan, the hemostatic and analgesic effect of this biopolymer should be
considered. Chitosan’s unique hemostatic properties are independent of the normal clotting
cascades [130]. In addition, chitosan can interact with neutrophils and macrophages and regulate
the re-epithelialization process and fibroplasia [131,132].
Howling et al. showed that chitosan enhanced fibroblast proliferation, and it seems that
the proliferative effect is related to the deacetylation degree of the chitosan. The samples with a
high deacetylation degree (84% and 86%) increased the mitogenic activity in fibroblasts, while
some samples of chitin had an antiproliferative effect [133]. Possibly, the mechanism by which
chitosan enhances fibroblast proliferation is by forming polyelectrolyte complexes with growth
factors, heparin, cytokines, or other proteins found at a wound site [134].
At physiological pH, the applications of chitosan are limited due to its poor solubility and
the fact that the drug delivery process is less efficient at pH below 6. Therefore, in order to
overcome these drawbacks, permanent changes can be introduced into the chitosan backbone.
Patrulea at al. developed a water-soluble O-carboxymethyl-N,N,N-trimethyl chitosan (CMTMC)
with a high degree of substitution (up to 46.6%), which did not exhibit significant toxicity
towards human dermal fibroblasts (viability > 80% at 1 and 0.5 mg/mL) [135]. A more recent
study published by Patrulea et al. showed that formulations based on carboxylated and
trimethylated chitosan (CMTMC) have great potential in wound-healing applications. Foams and
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hydrogels with a high concentration of RDG (arginyl-glycyl-aspartic acid)-functionalized

chitosan (3%) and hyaluronic acid (1.5%) not only showed no toxicity towards human dermal
fibroblasts but also promoted proliferation over 7 days and the migration of human dermal
fibroblasts [136].
Both hemostatic and analgesic features may be attributed to the positive charge of
chitosan. Given the fact that the red blood cells are negatively charged, they can interact with
cationic chitosan molecules; therefore, chitosan’s hemostatic effect relies on electrostatic
adhesion to blood cells [137]. The exact hemostatic mechanism of chitosan is yet to be fully
understood, but theories rely on plasma sorption, erythrocyte coagulation, and platelet activation
and aggregation [138]. The protonated amino groups of chitosan attract negatively charged red
blood cells, triggering the hemagglutination process [139]. Chitosan stimulates platelet
aggregation and adhesion by adsorbing plasma proteins and also signals thrombin, which is a
clotting promoter, enhancing the expression of glycoprotein IIb/IIIa (GPIIb/IIIa), a platelet
membrane receptor [140]. The analgesic effect of chitosan has been proven since 1984, when
Allan et al. reported that chitosan provided a cool and southing effect when applied on open
wounds [141]. Huang et al. tested the analgesic effect of chitosan and carboxymethyl chitosan on
scalded rats by monitoring the concentration of bradykinin and 5-hydroxytryptophan, potent
algogenic substances, using an enzyme-linked immunosorbent assay. Results showed that in the
case of carboxymethyl chitosan treatment, the concentrations of bradykinin and 5-
hydroxytryptophan were significantly lower than the control group (p < 0.05), but in the case of
chitosan treatment, the values were same as the control group (p > 0.05). Therefore, only
carboxymethyl chitosan showed an analgesic effect [142].
Chitosan’s structural characteristics are similar to glycosaminoglycans (GAGs), which
are components of the extracellular matrix (ECM); therefore, this feature recommends chitosan
use in skin tissue engineering [143]. The immobilization of bioactive molecules on the chitosan
matrix, such as cell-adhesive peptides, will stimulate the cell response and cell adhesion process.
In this regard, Karakeçili et al. have covalently immobilized mouse epidermal growth factor
(EGF) on chitosan membranes. The mouse EGF is responsible for fibroblast proliferation. The
results indicated that EGF has enhanced the proliferation of L929 mouse fibroblast cells [144].
Although there are published experimental data on how chitosan interacts with mammalian cells
involved in wound healing processes, there is still a lack of knowledge regarding innate sensing
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of chitosan and the immunological response towards this polymer [145]; as polysaccharides are
recognized by the cell receptors and depending on their size and structure, may induce immune
responses [146]. The fact that chitin and chitosan are not present in mammalian cells makes them
potentially recognizable targets for the innate immune system. However, chitooligosaccharides
could be recognized by mammalian chitinases and finally degraded [145]. Another study
revealed that chitosan, but not chitin, had triggered inflammatory responses by activating NLRP3
inflammasome in a phagocytosis-dependent manner. While chitosan was a potent NLRP3
inflammasome activator, acetylation of chitosan to chitin resulted in an almost complete loss of
activity [147]. Given the fact that chitosan is used in various biomedical applications, research
focusing on the immunological response is of great relevance.
5. FACTORS INFLUENCINGCHITOSAN’S ANTIMICROBIAL EFFECT

Many research articles describe the influence that different factors have on the
antimicrobial properties of chitosan. Some of the most important factors include molecular
weight and degree of deacetylation, pH, type of microorganism, microbial growth media
components, the source of the chitosan, and also its derivatives [19].
5.1. THE MOLECULAR WEIGHT AND DEGREE OF ACETYLATION
The molecular weight and degree of acetylation of chitosan define several properties of this
polymer [148]. The molecular weight of chitosan has a great influence on its biomedical
properties. Moreover, the degree of acetylation strongly influences its antimicrobial properties by
increasing its solubility and its positive charge. As shown in Figure 4, some physico-chemical
properties such as solubility, viscosity, and biocompatibility increase once the DA decreases,
while crystallinity and biodegradability are directly proportional to the DA. Biological properties
such as antimicrobial, analgesic, antioxidant, hemostatic, and mucoadhesive abilities increase
once the degree of acetylation is lower [149].
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Figure 4. Influence of degree of acetylation (DA) on chitosan physico-chemical and biological

properties.
Chitosan are often classified as high molecular weight (HMW), medium molecular
weight (MMW), and low molecular weight (LMW) [19], but the values for each type are not
well defined and the literature data is relative and sometimes even confusing. Published data
show that the inhibitory mechanism of chitosan against pathogen microorganisms is also related
to its molecular weight. Thus, while HMW chitosan forms a dense layer on the surface of the
cell, preventing nutrient uptake [89], the much smaller chitosan molecules seem to penetrate the
membrane and even bind to DNA and RNA [98,159]. The antibacterial effect is dependent on
both molecular weight (MW) and type of microorganism. Tin et al. tested the antimicrobial
activity of four different MW chitosan samples (LMW, MMW, HMW, and chitosan
oligosaccharide lactate) against Pseudomonas aeruginosa. The results showed that the MIC
(minimum inhibitory concentration) values were higher for chitooligosaccharides (4096 µg/mL)
compared to other chitosan samples (32 µg/mL) [160]. Another study performed by Batista et al.
illustrates the importance of MW on antimicrobial activity. They have tested two types of
chitosan (HMW and LMW) against E. coli (Gram-negative), L. casei, and multiresistant strain S.
aureus (both Gram-positive). Only MMW chitosan was antibacterially active, and only against E.
coli and L. casei, while S. aureus was resistant to both chitosan samples [161]. As shown in
5.2. pH EFFECT
The beneficial properties of this polymer depend strongly on its solubility, in either water
or other commonly used solvents. In its crystalline form, chitosan is soluble in aqueous solutions
only at a pH value under 7. Its solubility is due to the protonation of its amino groups under
acidic conditions. The main disadvantage of chitosan is its low solubility, and over the past
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years, there has been a growing interest in the chemical modification of this polymer in order to
improve its solubility [149,165,166]. At a pH value of approximately 6.0, chitosan carries a
positive charge, being able to interact electrostatically with various negatively charged
molecules. Therefore, the antimicrobial mechanism of chitosan is pH-dependent.
As described earlier in this paper, chitosan interacts with the negatively charged cell
membranes of bacteria and fungi, leading eventually to the death of the microbial cell [167].
Many studies show that chitosan’s antimicrobial activity is minimal at higher pH conditions.
Younes et al. demonstrated that when reducing pH values, the adsorption on bacterial cell
surfaces will increase, probably due to the highly positive charge on the chitosan polymer chain
[34]. On the contrary, at higher pH (above 6), chitosan will lose its charge, and due to the
deprotonation of its amino groups, it will precipitate from solution. Devlieghere et al. observed
during their study that native chitosan showed greater antifungal activity against Candida
lambica at a pH value of 4.0 rather than 6.0 [106]. Helander et al. also observed a greater
antimicrobial effect of chitosan at lower pH conditions, while the inhibitory activity decreased
with increasing pH [77]. Erdem et al. tested the influence of pH on the antimicrobial activity of
chitosan against two strains: S. aureus and Aeromonas hydrophila.
The pH values selected were 5.0, 6.0, 7.0, and 8.0. Chitosan showed an increased
inhibitory effect at lower pH (5–6) against both strains, reducing the viable cells from their initial
populations to 1.80–0.57 log CFU/mL for A. hydrophila and 1.70–1.00 log CFU/mL in the case
of S. aureus. In addition, they observed that unmodified chitosan does not present antimicrobial
activity at a pH of around 7.0 [168].
The results of Batista et al. are similar in that they observed the highest inhibitory effect
of chitosan against E. coli and Lactobacillus casei at a pH value of 5.7 (L. casei) and 5.5 (E. coli)
[161]. While testing the antifungal activity of LMW chitosan against several species of clinical
isolates of Candida sp., Alburquenque et al. observed an increased inhibitory effect at a pH value
of 4.0, with the MIC being up to 4 times lower than at neutral pH. Their theory was also based
on the fact that the increased antifungal activity is due to the protonation of the amino group of
chitosan units at pH 4.0, as the pKa of LMW chitosan is 6.3 [169].
5.3. SOURCE OF CHITOSAN
Researchers have already investigated and compared the antimicrobial effect of chitosan
extracted from different sources. Tajdini et al. compared the antimicrobial effect of shrimp
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chitosan with fungal chitosan against twelve bacterial and fungal species. The results showed
that fungal chitosan wasmore efficient against P. aeruginosa, E. coli, Candida glabrata, and C.
albicans in comparison to the shrimp chitosan, the MIC values being twice smaller for fungal
chitosan. Against the other microorganisms tested, the antimicrobial effect was similar, and MIC
values were the same for both types of chitosan, indicating that fungal chitosan can be an
effective and suitable alternative to shrimp chitosan [170]. Chien et al.’s study revealed that
chitosan extracted from shiitake mushrooms was more effective than shrimp chitosan against
some of the pathogenic microorganisms tested [98]. Tayel et al. published a study that showed
that chitosan extracted from Mucor rouxii DSM-1191 exhibited pronounced antifungal activity
against different Candida albicans strains. Four types of fungal chitosan were tested, and the
most active against the fungal strain had the highest deacetylation degree (94%) and the lowest
molecular weight (32 kDa). [162]. Moussa et al. also demonstrated that fungal chitosan
(deacetylation degree of 89.2% and a molecular weight of 2.4 × 104 Da) extracted from
Aspergillus niger mycelia had a high antimicrobial effect against two foodborne pathogens
(Salmonella typhimurium and Staphylococcus aureus) [171].
Extraction of chitosan from fungal sources has many advantages over crustacean sources:
(a) Eliminates the harsh chemical processes that may cause environmental pollution;
(b) The raw material can be available throughout the entire year, and there are no risks of heavy
metal contamination;
(c) The demineralization step is no longer necessary for the extraction of chitosan from fungal
mycelia;
(d) Different types of waste products can be used to grow fungal cultures, reducing marine
pollution by the removal of nitrogen, proteins, and other compounds from waste [107,117].
Moreover, as an alternative to crustacean-derived chitosan, fungal strains may be adapted to
industrial needs by applying genetic modifications in order to produce larger amounts of chitin
for further production of higher-quality chitosan [172].
However, some important advantages of chitosan extraction from crustacean waste are worth
mentioning. First, crustacean waste is an important source of pollution; therefore, obtaining
chitosan from this waste material will give value-added products. Furthermore, if enzymatic
extraction methods are used, the harsh environmental impact that chemical extraction methods
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involve will be reduced, and the result will be a well-defined chitosan, obtained by a controlled,
non-degradable process [173,174].
5.4. TYPE OF MICROORGANISM
It is a fact that chitosan possesses innate antimicrobial properties, and most studies show
that its antimicrobial properties depend on the type of microorganisms. Although most of the
results indicate that chitosan has a more pronounced antimicrobial effect on Gram-positive
bacteria rather than Gram-negative [175], the literature is contradictory, as other studies have
demonstrated the opposite [124,176]. For example, Fernandes et al. showed that the
antimicrobial effect of chitosan was dependent on the target microorganism as well as the
molecular weight. Thus, chitooligosaccharides had a higher antibacterial effect on Gram-
negative species (E. coli, Klebsiella pneumoniae, P aeruginosa, MIC values of 0.10%, 0.10%,
0.20%, respectively) compared with Gram-positive ones (S. aureus and S. epidermidis, MIC
values of 0.25% and 0.20%, respectively). In the case of Gram-positive bacteria, high-molecular-
weight chitosan exhibited a stronger antibacterial effect than chitooligosaccharides (MIC values
of 0.10% for both strains), while for Gram-negative bacteria, MIC values were higher (0.25% for
E. coli and K. pneumoniae and 0.5% for P. aeruginosa). Likewise, for C. albicans, the results
were rather similar to those detected for Gram-positive bacteria; chitooligosaccharides inhibited
fungal growth at an MIC value of 0.25%, while the MIC value for high-molecular chitosan was
0.15% [177].
The result of a study by Chung et al. showed that chitosan’s inhibitory effect was higher
against Gram-negative bacteria. Their hypothesis was related to the hydrophilicity of the
bacterial cell wall. The results obtained showed a greater hydrophilicity in Gram-negative
bacterial cell walls then Gram-positive ones; therefore, more negatively charged bacterial surfaA
recent study on the antimicrobial effect of chitosan against bacterial contamination on the surface
on Iranian banknotes shows that the inhibitory effect is stronger against Gram-positive bacteria,
and a possibility of avoiding bacterial contamination through banknotes could be their coating
with chitosan [179].
The components found in the growth media used to cultivate the microorganisms may
also influence the antimicrobial properties of chitosan. Devlieghere et al. demonstrated that
media food components could influence the effect of chitosan on microbial growth. For example,
0.005% chitosan was enough to inhibit Candida lambica growth, and small amounts of starch
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(1% w/v) did not affect chitosan antimicrobial properties. However, high amounts of starch (30%
w/v) significantly decreased the activity of chitosan against C. lambica. In addition, only 1% of
NaCl was enough to inhibit the antimicrobial activity of chitosan [106].ces had greater
interaction with positively charged chitosan [178].
5.5. CHITOSAN COMPLEXES
The main purpose of producing chitosan complexes with other compounds is to improve
its beneficial properties, most of all, its antibacterial effects, and accelerate wound healing.
Various studies present different complexes between chitosan and other natural or synthetic
compounds in order to improve its antimicrobial effect and accelerate the wound healing process.
For example, Liu et al. developed an antibacterial material based on chitosan oligosaccharide-N-
chlorokojic acid mannich base (COS-N-MB). The resulting COS-N-MB complex with
synergistic antibacterial effects showed excellent inhibitory activity against pathogenic bacteria.
The mechanism of action seems to rely on the adsorption to bacterial cell walls through
electrostatic interactions and chelating metal ions [125]. Biswas et al. developed two distinct
wound-dressing materials by loading porous chitosan/poly(vinyl alcohol) (CS/PVA) scaffolds
with Ag and Se using an in situ deposition method.
The Ag-CS complex showed significant inhibition of bacterial growth and reduction of
cytotoxicity to fibroblasts, while the Se-CS complex damaged bacterial cell membranes and
showed no toxicity towards fibroblasts [180]. Ma et al. observed that the incorporation of
glycerol enhances mechanical properties and maintains the good water vapor permeability and
wettability of chitosan-based membranes [181]. The study of Kaygusuz et al. demonstrated that
cerium ion–chitosan crosslinked alginate films show antibacterial activity against both Gram-
negative (E. coli) and Gram-positive bacteria (S. aureus) and also show high mechanic
resistance, flexibility, and UV protection [182]. Anjum et al. developed antimicrobial and scar-
preventive wound dressings by blending chitosan (CS), polyethylene glycol (PEG), and
polyvinyl pyrolidone (PVP) on a cotton fabric. The cotton fabric was used as a support layer for
the CS-PEG/PVP gel; PEG increased the mechanical properties, and polyvinyl pyrolidone (PVP)
helped hydrogel formation [183]. Wahid et al. prepared Carboxymethyl chitosan (CMCh)
supramolecular hydrogels cross-linked by metal ions (Ag+, Cu2+, and Zn2+) that showed
moldability, elasticity, high mechanical properties, and a rapid and facile gelation process
(within seconds). Moreover, the metal ions added to the CMCh membranes significantly
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enhanced antimicrobial activity (Zn-loaded CMCh membranes had higher efficiency against all
bacteria tested) [184].
Although chitosan’s antibacterial activity is lower compared with conventional antibiotics, its
unique chemical behavior, namely the presence of three reactive functional groups (an amino
group at the C-2 position, a primary OH group at the C-6 position, and secondary OH groups at
the C-3 position), make this polymer available for further chemical modification in order to
improve its antimicrobial activity [72,125].
6. CHITOSAN ANTIMICROBIAL
Wound Dressings Wound infections can severely affect the healing process, and if not
properly treated, they can lead to sepsis and even to patient death [185]. The human skin acts as a
barrier against external factors, but when injured, it loses its efficacy and becomes vulnerable to
pathogenic infections [186,187].
The ideal wound dressing should meet some requirements such as:
(1)representing a physical barrier that is permeable to oxygen but at the same time maintains or
provides a moist environment; (2) sterile and non-toxic and protective against microorganism
infections; (3) providing an appropriate tissue temperature to favor epidermal migration and
promote angiogenesis; (4) non-adherent to prevent traumatic removal after healing [188]. All the
above-mentioned properties are characteristic of an ideal wound dressing, but it is difficult for
one type of dressing the meet all these requirements.
In addition, it is preferable that wound dressings contribute to skin regeneration. In this
regard, a variety of wound-dressing materials have been developed either based on synthetic or
natural polymers. There are many major types of wound-dressing materials: fibers, gels,
membranes, films, sponges, and hydrocolloids. Although the terms “film” and “membranes”
could be use to describe the same type of wound dressing, referring to a device developed to
isolate a surface from its environment, we have chosen to use and describe the terms separately,
mainly due to the fact that the terms vary among authors [195–199]. Moreover, both terms are
used accordingly by experts from different fields; for example, the term “membrane” is
employed mainly in the biological and health sciences, while professionals from engineering
science use the term “film”. The distinction between the two terms can be made based on the
moisture content, as membranes are hydrate films while films are considered dried membranes
[200].
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6.1. CHITOSAN FIBERS

There are several common ways to produce chitosan fibers, the first published data being
reported in 1926 [90]. Fibers can be produced by a dry and wet spinning method using different
solvents. A more recent technique used by modern-day scientists is electrospinning (ESP). The
main advantage of this method is that it allows the fabrication of polymer nano- and microfibers,
depending on the processing conditions [228]. Porous fiber membranes have a large potential in
wound-healing applications due to their high porosity and the ability to mimic the skin’s
extracellular matrix.
Zhou et al. synthesized N,N,N-trimethyl chitosan (TMC) fibers with different degrees of
quaternization (DQ 19%, 25%, and 32%). TMC fibers showed higher antibacterial activity
against Gram-negative E. coli (>63%) and Gram-positive S. aureus(>99%) than chitosan fibers
(24%). In addition, TMC fibers showed no cytotoxicity toward mouse fibroblasts and the TMC
DQ of 25% significantly increased wound reepithelization compared to the control (chitosan
fibers). The results indicated that a high DQ increased antibacterial activity due to the quaternary
ammonium groups with permanent positive charges. The cationic charge is the key to the
binding process of the polymer to the negatively charged bacterial cell membrane [229].
6.2. CHITOSAN HYDROGELS
Hydrogels are 3D polymer networks that can absorb large amounts of water and are
moist, flexible, and soft materials with a wide range of applications in biomedical fields. The
high content of water and the mechanical properties of hydrogels make them compatible with
most living tissues, and they improve the healing process [127]. The hydrophilic gel keeps the
wound bed moist, providing a non-adherent environment, and their swelling capacity helps with
wound exudate absorption [230]. Another advantage of hydrogels is that they can follow the
geometry of the wound, assuring a good superficial contact, which can be very helpful even in
third-degree burns [165].
In addition to their main purpose (drug and/or growth factor delivery while speeding up
the healing process), hydrogels must also prevent the microbial infections that can develop in
this moist environment. Antimicrobials can be either covalently bound to the hydrogel network,
noncovalently encapsulated in hydrogels, or the hydrogel can possess inherent antimicrobial
properties. Amongst other antimicrobial polymer-based gels, chitosan-based hydrogels are of
great interest for the scientists [231].
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6.3. CHITOSAN MEMBRANES

Amongst the variety of wound dressings available, the porous membrane dressings are
considered to be the ones that meet most of the requirements of an ideal wound dressing [51].
However, the moist environment provided by membranes also facilitates pathogen proliferation,
thus it is highly necessary to impregnate the membranes with an antimicrobial agent.
The simplest method of developing chitosan membranes is a solution casting–evaporation
technique, by which chitosan is first solubilized in acetic acid solution [232]. The main drawback
of this method is that acetic acid, along with chemical cross-linkers, such as carbodiimide or
glutaraldehyde, among others, possess cytotoxic effect on mammalian cells; therefore, this is a
major limitation to wound healing. To overcome these disadvantages, Ma et al. developed
chitosan–glycerol membranes loaded with tetracycline hydrochloride (TH) and silver
sulfadiazine (AgSD) through a simple solution casting–evaporation method and evaluated the
bacterial growth inhibition against E. coli and S. aureus by measuring the inhibition zone
diameter. The difference was that they used chitosan floccule suspension instead of an acetic
acid solution. Thus, chitosan–glycerol membranes loaded with antibacterial agents not only
inhibited bacterial growth in a significant manner, but glycerol also enhanced the membranes’
mechanical properties, promoting wound healing [233].
6.4. CHITOSAN FILMS
Güne¸s et al. developed a chitosan-based film incorporating Hypericum perforatum oil
that proved to have an antimicrobial effect on E. coli and S. aureus. The results showed that the
antimicrobial effect increases with the concentration of essential oil incorporated into the
chitosan matrix. Thus, the diameter of the zone of inhibition of simple chitosan films against E.
coli was 2 cm compared to 2.9 cm in the case of chitosan films incorporated with 1.5% H.
perforatum oil. S. aureus proved to be more resistant than E. coli, with a diameter of the
inhibition zone of chitosan films with an essential oil concentration of 1.5% equal to 1.97 cm
[223].
There are numerous chitosan-based products on the market, and one example is the
HemCon® dressing, which is a chitosan acetate bandage that was designed as a hemostatic
dressing with antimicrobial properties. Burkatovskaya et al. investigated its effect on infected
and non-infected excisional wounds in mice. The wounds infected with Gram-negative species
(P. aeruginosa and Proteus mirabilis) and left untreated led to serious infections that caused the
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death of mice, but the chitosan acetate bandage applied on the infected area rapidly killed
bacteria [126].
6.5. CHITOSAN SPONGES
Sponges are foam-like, solid structures that can absorb large amounts of liquid due to
their high porosity. Besides possessing antimicrobial properties, chitosan sponges are widely
used as wound dressings, being able to absorb wound exudates and also enhancing tissue
regeneration [165].
Anisha et al. developed an antimicrobial chitosan–hyaluronic acid/nanosilver composite
sponge for the treatment of infected diabetic wounds. The sponges showed an antibacterial effect
against the major bacterial strains known to infect a wound (E. coli, S. aureus, P. aeruginosa, K.
pneumoniae, and even MRSA—Methicillin-resistant Staphylococcus aureus). However,
cytotoxicity tests showed that higher concentrations of nanosilver in the sponges decreased cell
viability [234]. Another study published by Shao et al. showed that silver-sulfadiazine-loaded
chitosan composite sponges had high porosity and excellent swelling behaviors and possessed a
broad spectrum of antibacterial activity (E. coli, C. albicans, S. aureus, and B. subtilis). The cell
viability results performed by 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium
bromide (MTT) viability assay and fluorescence staining method on HEK293 cell lines indicated
that the sponges had no significant cytotoxicity; therefore, CS/AgSD composite sponges have
potential applications as antimicrobial wound-dressing materials [235]. Pei et al. fabricated
AgNP-loaded silk fibroin (SF)/carboxymethyl chitosan (CMC) composite sponges that showed
effective antibacterial activity against S. aureus and P. aeruginosa.
The addition of CMC increased the water vapor transmission rate and improved the water
absorption capacity and retention ability of the sponge, all of these being important properties of
a wound dressing [236]. Mi and team developed a sponge-like asymmetric chitosan membrane
by an immersion–precipitation phase-inversion method. The sponge-like membranes showed
many advantages a wound dressing requires, such as excellent oxygen permeability, and
controlled evaporative water loss, promoted fluid drainage ability, and inhibited exogenous
microorganism invasion due to the inherent antimicrobial properties of chitosan. Moreover,
histological tests on rat wounds showed an increased epithelialization rate and well-organized
deposition of collagen in the dermis [237].
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6.6. CHITOSAN HYDROCOLLOIDS

The composition of a hydrocolloid consists of an external layer of polyurethane and one
internal layer composed of hydrophilic colloidal particles (carboxymethylcellulose, gelatin,
pectin). A hydrocolloid dressing will absorb wound exudate due to its internal layer of gelatin,
pectin, and carboxymethylcellulose. The external polyurethane layer will seal the wound, not
only enabling gas exchange and preventing external contamination of the wound, but also
maintaining an acidic pH at the wound bed, facilitating autolytic debridement [90,204]. In
addition, they promote angiogenesis, increase the number of fibroblasts and the number of
collagen fibers, and enhance the production of granulation tissue, all of each are very important
steps in the healing process [204].
Hiranpattanakul et al. developed a chitin/chitosan hydrocolloid (CCH) wound dressing,
and properties like their water absorption, enzymatic degradation, and antibacterial activity
against Gram-positive and Gram-negative bacteria were evaluated, as well as their
biocompatibility with the L929 cell line. The CCH showed antibacterial activity against E. coli
and S. aureus and showed no cytotoxicity against the L929 fibroblasts [231]. Yanagibayashi et
al. developed another functionalized wound dressing to stimulate wound healing in diabetic
mice. The hydrocolloid composed of alginate, chitin/chitosan, and fucoidan (ACF-HS) had
important properties like adherence, ease of application and removal, providing a moist
environment, and absorbing exudate. Moreover, the histological examinations of the wounds
showed advanced tissue and capillary formations, starting on the 4th day of treatment [238].
The multitude of wound dressings already existing on the market, from sponges to
granules, gels, and sprays, only prove that chitosan is a promising polymer for wound-healing
applications, being versatile and suitable for many types of wounds. Although the majority of
chitosan-based wound dressings, presented below, are hemostatic dressings developed to stop
moderate to severebleeding, they also have specific features that direct them to certain types of
wounds. For example, Chitoderm® plus is a wound dressing based on a strong non-adhesive
superabsorber, with chitosan coating, that has bacteriostatic action and actively accelerates
healing processes. The manufacturer recommends it for numerous types of wounds, from acute
to chronic, exudating, contaminated wounds, venous leg ulcers, diabetic, and first- and second-
degree burns [239]. Another product, Celox™, is a hemostatic chitosan-based wound dressing
that clots hypothermic blood and blood treated with blood-thinning drugs, can be easily removed
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from the wound site, and the residual material is naturally absorbed by the body [240]. The
manufacturers of Opticell gelling fibers had an interesting approach in developing this chitosan-
based dressing. Not only can the dressing be worn up to seven days, but the chitosan-based
Cytoform technology allows the absorbent fibers within the dressing to transform into a gel.
Indicated for the control of minor bleeding, the gelling action helps manage drainage and the
removal of dead, damaged tissue, trapping it until the later removal of the dressing [241]. A
similar approach is seen for the LQD spray dressing, which contains chitosan as active
ingredient, that has been modified to have the highest degree of deacetylation of any chitosan
product and a much higher positive charge. The LQD spray is applied as a spray and within two
minutes of application, it forms a chitosan membrane over the wound. This dressing, which is
indicated for the external treatment of chronic and acute wounds and superficial and partial
thickness burns, providing physical protection and suppressing the secretion of proinflamatory
cytokines, contributing to pain relief, has a hemostatic and antibacterial effect [242].
The current regulatory status of chitosan given by the US FDA (United States Food and
Drug Administration) is defined as GRAS (generally recognized as safe) for wound-dressing
applications [243–245] and also tissue engineering [246,247]. Moreover, chitosan has been
approved as a dietary supplement in Japan, Italy, and Finland [248]. Therefore, there is a
growing number of chitosan wound dressings that meet the requirements of various types of
wounds, and due to the advantages provided by this polymer, the use of chitosan as a wound-
dressing material will only be increasing in the future.
7. CONCLUSIONS
There is no doubt that chitosan possesses important features that can be very helpful in
solving modern-day challenges regarding various areas of interest such as biomedicine,
pharmaceutical, cosmetic, and food industries, wastewater treatment, and agricultural pest
management.
Chitosan may be one of the most important biopolymers that can be used to produce
wound dressings that not only accelerate the wound healing process but also prevent infection of
the wound, a necessary step in the wound healing process. The mechanism of action against
microbial cells is not entirely defined, but numerous studies have presented similar results;
therefore, researchers have formulated some hypotheses by which chitosan can inhibit microbial
growth:
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(1) Electrostatic interactions occur between cationic chitosan and anionic molecules at the
microbial cell surface, which may lead to cell wall disruption and intracellular components
leakage;
(2) LMW chitosan can penetrate the cell membrane and interact with DNA, thereby interfering
with protein synthesis processes;
(3) Another mechanism of action for chitosan would be the chelation of nutrients and essential
metals that are fundamental to cell stability.
Used as a wound dressing, chitosan stimulates the natural healing process and has no toxicity to
the mammalian cell. Being biocompatible, chitosan has a known metabolic pathway. Once it
enters the human body, it can be hydrolyzed by enzymes, such as lysozyme, present in mucus,
tears, and saliva, to an important sugar, glucosamine, that it is already present in the human
body.
The existence of numerous chitosan-based wound dressings on the market shows the importance
of this biopolymer in accelerating the wound healing process, and we may consider chitosan as a
cost-effective solution not only for the treatment of acute wounds but also in the case of severe
chronic wounds, such as diabetic ulcers.
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Spotz, Z.; Hebeish, A. Novel chitin/chitosan-glucan wound dressing: Isolation,
characterization, antibacterial activity and wound healing properties. Int. J. Pharm. 2016,
240. 220. Han, F.; Dong, Y.; Song, A.; Yin, R.; Li, S. Alginate/chitosan based bi-layer
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241. 221. Chen, H.; Xing, X.; Tan, H.; Jia, Y.; Zhou, T.; Chen, Y.; Ling, Z.; Hu, X.
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242. 222. Mishra, S.K.; Mary, D.S.; Kannan, S. Copper incorporated microporous
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ISBN: 978-81-948129-1-3 Page 375

A COMPREHENSIVE REVIEW ON NANOPHOTONICS AND ARTIFICIAL

INTELLIGENCE
Sridhar Arelli1 and Srivenkaiahppalaswamy B2
1
Department of Physics, OPJS UNIVERSITY, Churu, Rajasthan., Mail: usri89760@gmail.com
2
Department of Computer Science, Adikavi Nannaya University MSN Campus, Kakinada East
Godavari, Andhra Pradesh., Email: bvaswamy@gmail.com
ABSTRACT:
Nanophotonics has been an active research field over the past two decades, triggered by
the rising interests in exploring new physics and technologies with light at the nanoscale. As the
demands of performance and integration level keep increasing, the design and optimization of
nanophotonic devices become computationally expensive and time-inefficient. Advanced
computational methods and artificial intelligence, especially its subfield of machine learning,
have led to revolutionary development in many applications, such as web searches, computer
vision, and speech/image recognition. The complex models and algorithms help to exploit the
enormous parameter space in a highly efficient way. In this review, we summarize the recent
advances on the emerging field where nanophotonics and machine learning blend. We provide an
overview of different computational methods, with the focus on deep learning, for the
nanophotonic inverse design. The implementation of deep neural networks with photonic
platforms is also discussed. This review aims at sketching an illustration of the nanophotonic
design with machine learning and giving a perspective on the future tasks.
KEYWORDS: deep learning; Nano photonic neural networks; inverse design; optimization.
1. INTRODUCTION
Nanophotonics studies light and its interactions with matters at the nanoscale [1]. Over
the past decades, it has received rapidly growing interest and become an active research field that
involves both fundamental studies and numerous applications [2,3]. Nanophotonics comprises
several subdomains, including photonic crystals (PhCs) [4], plasmonics [5],
metamaterials/metasurfaces [6-8], and other structured materials that can perform photonic
functionalities [9]. Despite the different underlying mechanisms, configurations, materials and so
ISBN: 978-81-948129-1-3 Page 376
forth, traditionally, the design of nanophotonic devices relies on physics-inspired methods.

Human knowledge, such as the physical insights revealed by the study of simple systems, the
experience obtained from previous practice, and the intuitive reasoning, provide guidelines to the
design process. For example, knowing that an elongated nanoparticle responds more strongly to
the incident electric fields that are polarized along its long axis, and that a ring-like structure
supports magnetic resonances if the incident magnetic fields are perpendicular to the plane on
which the ring lies, the combination of metallic wires and split ring resonators (SRRs) was
proposed to demonstrate the negative index of refraction [10], a milestone of metamaterials. The
initial designs are usually examined by simulations solving the Maxwell’s equations, but they are
less likely to match the desired performance directly. Therefore, adjustments to a handful of
parameters and re-evaluation by simulations need to be conducted repeatedly to approach the
target. While remarkable success has been accomplished using this scheme, the trial-and-error
procedure becomes computationally costly and time-inefficient due to the continuously
increasing complexity of the nanophotonic devices.
Inverse design tackles the design task in a different manner [11]. Without the need of
physical principles for the initial guess, intended photonic functionalities are obtained by
optimization in the design parameter space, which, based on advanced algorithms and combined
simulations, seeks a solution that minimizes (or maximizes) an objective or fitness function
related to the target. In relation to solving the direct problems, optimization-based methods
require comparable computation power and time. Nevertheless, they allow one to search in the
full parameter space and find designs that are non-intuitive but with optimal performance.
The recent blossoming of artificial intelligence (AI), especially the subfield of machine
learning has revolutionized many realms of science and engineering, such as computer vision
[12], speech recognition [13], and strategy making [14], etc. Inspired by the biological neural
networks, artificial neural networks have dramatically changed the paradigm of data processing
and powered the development of algorithms that can “learn” from data and perform
functionalities to complete complex tasks [15]. The associated technique of deep learning is thus
considered a promising candidate for the inverse design of new materials [16-18], drugs [19],
and nanophotonic devices [20-22] (Figure 1). In general, the role of deep learning in
nanophotonic design is also to search the parameter space for a best fit of the target. But unlike
optimization-based methods doing this for every task, which makes simulations recurrent efforts,
ISBN: 978-81-948129-1-3 Page 377
deep learning algorithms are able to navigate in a smarter way by learning from a large dataset so
that a solution can be found almost instantaneously after the learning phase. Without loss of
design flexibility, this data-driven scheme markedly shortens the overall computation time when
a common database is available for a group of applications. On the other hand, nanophotonic
circuits that process coherent light are naturally suitable to build systems compatible with the
framework of neural networks [23], while the speed and energy efficiency can be much higher
than those of their electronic counterparts. Therefore, the application between deep learning and
nanophotonics is not one-way but interactive. As their blending is just beginning, it will be
timely and beneficial to present an overview on this emerging field, from which interested
readers can get a general idea and determine the directions of future research [24]. We notice that
some related techniques, such as topology optimization [25], inverse design [11], neuromorphic
photonics [26-29], and reservoir computing [30] have been discussed by several recent review
articles. Thus, we would also direct the readers to these references if interested.
The present manuscript is organized as follows. Section 2 summarizes the recent progress
in nanophotonic inverse design based on optimization. Popular techniques as well as
representative examples will be introduced. In section 3, we start by explaining the concept of
deep neural networks (DNNs). Important applications in designing novel devices, discovering
new phenomena, and revealing underlying mechanisms are then discussed with details. Section 4
is dedicated to the efforts to perform deep learning with nanophotonic circuits and optical
materials as hardware. Experimental results and theoretical models for all-optical deep learning
makes this topic extremely attractive and promising.
2. NANOPHOTONIC DESIGN BASED ON OPTIMIZATION TECHNIQUES
Computation-wise there are many different ways to solve a photonic design problem,
either direct or inverse, whereas the basis of any design strategy is that the optical properties of a
given structure can be modelled with enough accuracy. For this purpose, a variety of
computational tools have been developed, such as the finite-difference time-domain (FDTD)
method, finite element method (FEM), boundary element method (BEM), discrete dipole
approximation (DDA), and rigorous coupled wave analysis (RCWA), etc. Despite their own pros
and cons in fitting different applications, these approaches solve the governing equations of light
waves, i.e., Maxwell’s equations. The simulated results are evaluated by the designer or an
ISBN: 978-81-948129-1-3 Page 378

algorithm for optimization, and the updated structure is sent back to the solver for the next cycle
of simulation and optimization until the specified performance is reached.
To date, most popular algorithms used in the inverse design can be categorized into two
groups: the evolutionary method, such as genetic algorithm [31,32] and particle swarming
optimization [33-35], and the gradient-based method, including topology optimization [36,37],
steepest descent, and so forth. Other approaches based on heuristics (simulated annealing
[38,39]) or nonlinear search have also been used. The main advantage of using these techniques
over the traditional physics-inspired scheme is that it opens up the full parameter space and many
non-intuitive designs can be obtained with optimal performance. In this section, we summarize
the recent advances on nanophotonic design based on computational methods, primarily
optimization techniques. Due to the complex intersections of the many applications and
algorithms, priority will be given to the similarity between applications when the selected
examples are grouped, while different design methods will be introduced the first time they
appear in the text unless otherwise specified.
The earliest application of computational methods in nanophotonic inverse design dates
back to the late 1990s, with the attempts to optimize the performance of dielectric waveguides
[40] and to engineer the bandgaps of PhCs [41]. Since then, continuous progress has been made
along these lines [42-53], and some previously unattainable functionalities have been made
possible by using advanced algorithms and hardware [54-72]. Among the pioneers who
introduced various computational techniques into nanophotonic inverse design, Sigmund and
coworkers conducted a systematic study using the tool of topology optimization, which was
originally developed for structural design [36,37] but has been applied to many other
applications [25].
In topology optimization, the entire design domain is discretized into pixels, each being a
design variable that represents the material property at that point. The total number of variables
can thus be very large for a complex design task, and the structures are not restricted to any
certain class of geometries. The iterative optimization procedure consists of repeated simulations
and updates of the material distribution based on gradient computation. The latter is essential;
otherwise the efficiency decreases dramatically, given the many design variables in topology
optimization [73]. Figure 2(A) exemplifies this technique with a PhC Z-bend [74]. It is well-
known that sharp bends in a waveguide will cause significant bending loss and poor
ISBN: 978-81-948129-1-3 Page 379
transmission. Conventional optimization methods that are not free from geometric constraints
solve the problem by adjusting the hole sizes and disturbing the lattice in the whole bending area.
Topology optimization, in contrast, is shown to find with higher efficiency an optimized solution
with only five holes being reshaped on the outer part of each bend. Despite the slight
discrepancies between the designed pattern and fabricated structure, nearly 10 dB higher
transmission was experimentally achieved for a bandwidth of over 200 nm. In this specific
problem, the broadband property was obtained by optimizations at a single wavelength.
Nevertheless, any number of wavelengths can be used simultaneously to fit any desired spectra.
In addition to waveguide bends, devices with increasing complexity, such as mode converters
[75] and beam splitters [55,63], have been reported by the same group. Figures 2(B) and 2(C)
show two prototypes of beam splitters designed by different approaches. In the first example,
Piggott et al. demonstrated multi-channel wavelength splitting [76].
The specifications of this design task are the conversion efficiencies between the input and
output modes at discrete wavelengths, and two different methods were employed sequentially to
find the solution. At the starting point, an “objective first” strategy was adopted to take an initial
guess of the structure [77], which first constrained the mode profiles to satisfy the target
performance but allowed Maxwell’s equations to be violated, and then minimized this violation
with an optimization algorithm. Next, for fine tuning of the structure, the steepest (gradient)
descent method was applied by computing the gradient of the performance metric to find its local
minimum [78]. This process was under the constraint of Maxwell’s equations while the
permittivity was still allowed to vary continuously. The resulting layout was a complex gradient-
index (GRIN) pattern with the refractive index ranging from 1 (nair) to 3.49 (nSi). After
converting this pattern to a binary level-set representation [79], by which the material at each
position can only be air or silicon, the design was optimized again using the steepest descent
method for performance and bandwidth optimization around 1300 and 1550 nm wavelengths.
The whole design process took ~36 hours using a graphics processing unit (GPU) accelerated
FDTD solver.
Another design of nanophotonic beam splitters was showcased by Shen et al. [80] In this
example, an unpolarized input mode is split to transverse magnetic (TM) and transverse electric
(TE) components that exit the device at two different output ports. The design was based on a
direct binary search (DBS) algorithm, which differs from the gradient-based methods. In brief,
ISBN: 978-81-948129-1-3 Page 380
an area of 2.4 × 2.4 μm2 was first discretized into 20 × 20 pixels. Each pixel has a size of 120 ×
120 nm2 and represents a silicon pillar or a void, denoted by state 1 or 0. The thickness of the
device was also discretized with a step of 10 nm. A figure of merit (FOM) for optimization was
then defined as the average transmission efficiency for the TM and TE modes. Following a
random sequence, the state of the pixels was switched and FOM was calculated. If FOM was
improved after a switch, that pixel would retain the new state; otherwise it kept the original
value. After all the pixels were addressed, a similar optimization was applied to the device
thickness by changing its value to the adjacent states (±10 nm). Walking through the 400 pixels
and making a slight adjustment to the device thickness completed one iteration. The optimization
was terminated when the improvement of FOM was smaller than a threshold after an iteration or
the maximum iteration number was reached, which took ~140 hours. As a non-gradient
approach, the DBS is computationally intensive and becomes less efficient when the number of
design variables increases [73].
To maintain the computation time within an acceptable range, parallelizing the algorithm
and using larger clusters of processors would be necessary [81]. Figure 2(C) reports the design,
experimental and simulation results, showing reasonably good agreement. On-chip devices form
a class of applications suitable for inverse design. Besides beam splitters, optical diodes that
perform asymmetric spatial mode conversion [70, 71] (Figure 2(D)) and reflectors [82] (Figure
2(E)) have also been demonstrated using different algorithms.
Flat optics is another class of applications powered by optimization methods [7,8]. Having
resonant elements arranged at an interface or in a few layers to carry out functionalities,
metasurfaces and metalenses contain many variables to be carefully determined in the design
process, and the problem could be larger-scale if the structure is aperiodic. For the design of
dielectric metasurfaces, topology optimization could be a well-suited option [25]. Starting from a
random, continuous refractive index distribution bounded by the indices of air and the dielectric
(nd), structures satisfying the desired performance but with binary indices 1 and nd can be
achieved by using gradient-based optimization algorithms. Sell et al. reported an approach for
designing periodic silicon metasurfaces with multiwavelength functionalities [83-86]. When the
target was set to deflect light of discrete wavelengths to different diffraction orders, an FOM
involving all the diffraction efficiencies was defined for optimization. By using an adjoint-based
method [87], i.e., solving the forward diffraction problem and an adjoint problem that reverses
ISBN: 978-81-948129-1-3 Page 381
the incidence to the target diffraction order directions as one iteration, the gradient of FOM can
be calculated, based on which the refractive indices at each point received a small adjustment
towards nair or nSi. The iteration continued until a binary profile was obtained, as shown in
Figure 3(A). Similar procedures can be used to tackle aperiodic and multilayer structures
and to achieve different functionalities, such as wavefront manipulation, polarization control, and
beam shaping. Although considerable efforts have been devoted to alleviating the aberration in
metasurfaces, success was only attained in eliminating the chromatic aberration [88]. The
suppression of angular and off-axis aberrations in single-layer metasurfaces is fundamentally
impossible, whereas the design of multi-layer structures with an angle-dependent phase profile is
practically challenging for traditional approaches. Taking advantage of topology optimization,
Lin et al. designed a two-dimensional (2D) metalens that is free of angular aberration [89].
Figure 3(B) depicts the layout, which is symmetric but aperiodic, comprising five layers of
silicon gratings embedded in an alumina background. FDTD simulations revealed that at the
three target off-axis angles of incidence, light was all focused on the focal plane following the
identical diffraction limit, as shown in Figure 3(C).
Evolutionary algorithms are also widely used in metasurface design. The general strategy
is to maximize a fitness function by repeatedly evolving a population of candidate solutions with
sequential application of selection, crossover, and mutation, etc. One possible framework
consisting of four steps is illustrated in Figure 4(A), which was adapted by Huntington et al. for
optimizing a lattice of circular holes in a metal film to achieve unique focal properties [90]. The
design began with the generation of the initial population (step 0), a group of 600 randomly
created binary patterns. Because the lattice consists of 33 × 33 holes, each denoted as 1/0 when
the hole is open/closed, in total there are 21089 possible arrangements. The field distribution for
an arbitrary profile can be calculated by adding up the pre-stored complex fields from each
individual hole. Compared with simulating the entire structure, this scheme significantly
increases the efficiency. For a specification of the focal behavior, a fitness function can be
defined accordingly, which is maximized when the far-field intensity satisfies the target
functionality [91]. In step 1, each member in the initial population was evaluated by the fitness
function. The population was then sorted in step 2 by fitness, and the best-fit individuals were
selected in step 3 to create a new generation of the population through a combination of
crossover and mutation. The design cycle continued until a convergence condition was reached
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(step 4). In Ref. [90], after optimizing the parameters of the algorithm, e.g., the population size
and mutation rate, a specified design task can be finished in ~210 generations within 30 minutes.
But in general, since the optical properties of each individual structure is simulated rather
than pre-stored, the design would take a much longer time. In fact, the high computational cost
for large-scale design is an important limitation of evolutionary methods. Figure 4(B) shows a
lattice which exhibits five focal points arranged in a T-shape. In addition to the 1/0 state, the
holes were further encoded by their sizes. Although the same pattern can be produced with a
fixed hole size, the design with three different hole sizes improved the diffraction efficiency
from 55% to ~74%. Not only the far-field patterns but also the near-field intensity can be
engineered using this technique. Fitchtner et al. studied the near-field enhancement in a
checkerboard-type structure [92], which was optimized using an evolutionary algorithm. The
results led to a novel “matrix nanoantenna” structure that can provide a two-fold near-field
enhancement compared with a dipole antenna. Analysis of a reduced model in Figure 4(C),
which retains the important structural features of the fittest design, revealed that the enhancement
is caused by the complex interplay between a fundamental split-ring mode and a dipole mode in
the two arms.
Moreover, it is shown that by connecting nanobars to the ends of an SRR, the fundamental
resonance can be shifted from near-infrared into the visible regime. Tuning the resonances of
nanoantennas further enables color generation [93]. With the assistance of evolutionary
algorithms combined with an electrodynamic solver based on the Green’s dyadic function,
Wiecha et al. demonstrated polarization-dependent color pixels [94]. Figure 4(D) shows the
gallery of the designed silicon nanoantennas, which were optimized to have maximized
scattering at 550 nm for incident polarization along the x-axis and at various wavelengths for
incident polarization along the y-axis. Each pixel may have multiple elements and interestingly,
as the target wavelength increases, these elements tend to merge together. The polarization-
filtered dark-field spectra and images for orthogonal polarizations are compared in Figure 4(E),
showing reasonable agreement with the simulated results. The design methods can also be
combined with fabrications. Lee et al. integrated the processing steps of wrinkle lithography with
the concurrent design procedure of quasi-random light-trapping nanostructures for absorption
enhancement [95]. Specifically, the processing patterns were represented statistically by the
Fourier spectral density functions [96], which used only three variables to connect the structure
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and the optical property, making the problem solvable for a genetic algorithm. Figure 4(F)
depicts the optimization history of the averaged absorption enhancement by the designed
structures. After ~150 iterations the search gradually converged, resulting in an enhancement of
4.7 over the weakly absorbing interval of amorphous silicon from 800 to 1200 nm.
The suppression of light scattering by an object is a topic of broad interest. In recent years,
progress has been made with both forward design methods, such as transformation optics [97-
101] and scattering cancellation [102], and inverse design approaches [103]. Genetic algorithms
are usually adopted for optimizing a multilayer particle or cylinder to achieve omnidirectional
scattering reduction [104,105], while topology optimization is more practically associated with
designing bidirectional cloaks or resonators that can be realized by a low-index material [106-
112], although in theory it can work for any imaginable objective function [106,113]. Figure 5
summarizes a few examples based on topology optimization. Since the design methodology does
not differ much from the examples above, we will not proceed further to the details.
Lastly, we briefly outline a few other computational methods and applications. In addition
to device design, optimization algorithms have been used to explore new physics. In the study of
optical tweezers and optical manipulation [114-118], traditionally attention was paid to the
optimization of particle geometry and multiplexed optical traps. Lee et al. applied constrained
optimization [119], a derivative-free algorithm, together with a BEM solver to maximize the
optical torque on a gold nanotriangle [120] (Figure 6(A)). At the dipole and quadrupole
resonance wavelengths of the particle, a large portion of 2000 random initial illumination
conditions resulted in over 5-fold enhancement of optical torque per intensity, compared with
that from a standard circularly polarized planewave incidence. The optimal design at the
quadrupole resonance could even lead to a 20-fold improvement, as revealed in Figure 6(B). This
result provides new insights into the optical manipulation of objects with structured light and the
computational framework can be generalized to opto-mechanical applications. Lin et al.
demonstrated, based on topology optimization, that the third-order Dirac points formed by the
accidental degeneracy of modes belonging to three different symmetry representations can be
realized in inverse-designed PhCs [121] (Figure 6(C)). Moreover, the third-order exceptional
points (EP3) can be created by introducing a small loss term, giving rising to strong
modifications in the local density of states (LDOS) and potential connections to topological
photonics [122].
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Topology optimization is not the only technique compatible with the binary representation
of materials. Other algorithms, such as simulated annealing and particle swarm optimization,
have also been used in the optimization of nanostructures. Simulated annealing mimics the
process of heating and controlled cooling of a solid for recrystallization in metallurgy [39]. At
each iteration of the search, the algorithm keeps every better solution, and, by choosing a
temperature-dependent acceptance function, allows with a slowly decreasing probability some
worse solutions to stay in the pool. Therefore, this strategy largely avoids being trapped in local
minima and statistically guarantees finding a good solution, but meanwhile, its efficiency is
lower than gradient-based methods. Figure 6(D) shows the design of a binary plasmonic
structure composed of pixelated grooves [123]. Complex interference patterns of surface
plasmonpolaritons (SPPs) can be generated, showing the potential as plasmonic couplers.
Particle swarm optimization works based on the movements of a population of candidate
solutions (particles) in the search space. During optimization, the initially randomly distributed
particles continue moving towards the then-current optimum particle in the swarm, until a certain
termination criterion is reached [33]. Figure 6(E) reports the imaging of subwavelength holes by
a binary super-oscillatory lens [123], which was fabricated on a 100-nm-thick aluminum film on
glass and mounted to a microscope lens. The structure creates a delicate balance of the
interference of a large number of diffracted beams, ensuring the hotspot is very sensitive to the
presence of small objects. In addition to imaging, other reported applications include field
enhancement engineering [125], waveguide design [126], and color filters [127], etc.
3. NANOPHOTONICS ENABLED BY DEEP LEARNING
Deep learning, as a subfield of machine learning and AI, has attracted increasing
attention due to its great success in computer vision [12] and speech recognition [13] and its
astonishing progress in various applications such as strategy making [14]. Recently, owing to its
extraordinary capability in finding solutions from an enormous parameter space, researchers
have started using deep learning in drug discovery [19], materials design [16,18,128],
microscopy and spectroscopy [129-134], and other physics-related domains [135-137]. Among
all these attempts, nanophotonics turns out to be a unique field, because it not only benefits from
deep learning for the inverse design of advanced devices and performance improvement of
existing techniques, but can also give feedback, providing platforms to implement deep learning
algorithms that can operate at the speed of light and with low energy consumption. In the
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following, we discuss how the field of nanophotonics is actively interacting with the emerging
technique of deep learning. Specifically, the recent advances in applying deep learning for
nanophotonic design will be reviewed in this section, and the optical implementations of neural
networks are left to the next section.
We start with a brief introduction on some basic concepts about deep neural networks
(DNNs). Figure 7 illustrates the typical architecture of a DNN consisting of multiple processing
layers, including an input layer at the bottom, an output layer on the top, and at least one hidden
layer (but usually more) in between. Each circle in the diagram represents an artificial neuron,
which is connected to other neurons in the neighboring layers by different weight values subject
to learning. The input and output layers both have a fixed number of neurons or units,
determined by the size of the feeding data (for the input layer) and the task of the DNN (for the
output layer). In nanophotonic applications, they could correspond respectively to, e.g., the
design parameters of a nanophotonic structure and its optical properties or vice versa. The hidden
layers establish a nonlinear mapping between the input and output via training, from which very
abstract relationships can be discovered to make predictions on the optical properties of given
nanostructures and determine the design parameters for desired performance.
What lies at the very root of DNNs is the organization of the neurons. Specifically,
enabled by the use of the backpropagation algorithm, a unique feature of DNNs is that data can
be transformed bidirectionally through the network between the input and output layers.
Conducting deep learning is thus divided into two processes, the forward inference and the
training based on backpropagation, as sketched in Figure 7. In general, the computation of DNNs
is achieved by matrix multiplications. In the forward inference, starting from the input layer, the
neurons carrying input data form a vector X. Under full connectivity, each neuron xi in X is
connected to each neuron yj in the first hidden layer, Y 1 , by a weight wij. An initial value zj =
∑wij∙xi is given as a weighted combination of the neuron values from the previous layer. It is
then essential to apply a nonlinear activation function f to zj; its importance will be explained
shortly later. Now the value of neurons in the next layer Y 1 is rectified to yj = f(∑wij∙xi) or
simply Y 1 = f(W∙X), an expression of matrix multiplication. For the forward inference, each
next layer Y l+1 is connected to the previous layer Y l by a similar weight matrix, and this
operation is repeated for all layers until arriving at the output layer Y L for a network with L
layers, which gives the first guess of the target tk.
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Because the initial values of the weights wij are usually randomly chosen or at least not
well suited for the problem to be solved, this guess is very likely far away from the correct
answer, giving a large error. The training process works in the reverse direction. Based on the
error at each output neuron, a cost function C can be defined, which is minimized when the result
of the forward inference, Y L , is equal to the real answer. The gradient of the cost function can
be calculated by the partial derivative of the cost with respect to each weight variable, ∂C/∂wij.
Using the chain rule of derivatives, this can be expressed by ∂C/∂wij =
(∂zj/∂wij)(∂yj/∂zj)(∂C/∂yj). When calculating the final term for the weights connected to the
output layer, it is simple as the cost is directly a function of the values at those neurons. When
doing the same for previous layers, the derivative of the cost with respect to each neuron’s value
is a weighted sum of multiple errors, because the neuron is connected by multiple routes to all
the neurons in the output layer. Hence, the backpropagation is meant to efficiently calculate the
partial derivatives and how the error propagates through each layer. Finally, each weight is
adjusted by the partial derivative of the cost with respect to that weight, further scaled by a factor
η called the learning rate, Δw = −η∙∂C/∂w. Sometimes a stochastic factor is also included. This
weight update is how the network learns. The process of feeding data in, calculating a prediction
through the forward inference, calculating the cost by comparing it to the true values of that
training data, and calculating the gradient of the cost and adjusting each weight value is repeated
for many times. With a sufficiently large amount of training data, the performance of the DNN
can be continually improved.
While the basic principle of deep learning is summarized above, the actual
implementation of DNNs is much more complicated and contains many subtle problems, such as
the choice of training data, the cost function, network depth, initial weights, and the learning rate,
etc. Since these detailed techniques are not among the focus of this article, interested readers
may refer to some latest topical reviews or books, e.g. Ref. [138]. Before proceeding to the
applications in nanophotonic design, two concepts that will appear in the later discussion deserve
a glance. First, the nonlinear function f, or termed as activation function or transfer function,
plays an important role in DNNs. Compared with linear functions, nonlinearity allows a network
to tackle more abstract representation and learn much faster with fewer neurons. Popular choices
of the nonlinear function include the logistic functions, the hyperbolic tangents, and the rectified
linear units (ReLUs), etc. Second, the organization of neurons varies. Besides the fully connected
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network in Figure 7, another widely used architecture is the convolutional neural network (CNN
or ConvNet). In such structures, data flow in the form of multiple planes, and it is a filter or
kernel consisting of a small array of weights that connects the input and output planes. With this
change, CNNs contain much fewer connections than standard models with a similar depth and
are thus easier to train, while their theoretical best performance only decreases slightly [12]. This
characteristic is highly desirable when processing high dimensional data, such as images and
videos.
Applying deep learning algorithms to the nanophotonic inverse design introduces
remarkable design flexibility that can go far beyond that of conventional methods based on an
intuitive initial guess and many cycles of trial-and-error modeling, fabrication, and
characterization. It also enables, without recurrent efforts in conducting time-consuming
simulations, fast prediction of complex optical properties of nanostructures with irregular shapes
and intricate architectures. A bidirectional DNN that can achieve both the design and
characterization of plasmonicmetasurfaces was first reported by Malkiel and coworkers [20]. For
the implementation, two standard DNNs were used to perform the inverse design and spectra
prediction tasks for arrays of “H”-shaped gold nanostructures. Instead of training two networks
separately and composing them afterwards, it is shown that combining the networks during
training is more effective and helps to avoid unstable processing. The full structure of the
combined network is shown in Figure 8(A). A geometry-predicting-network (GPN) is used to
solve the inverse problem, for which the training data comprise two spectra for orthogonal linear
polarization excitations and the dispersive material properties. These three groups of data were
fed separately and in parallel into three DNNs before they join a larger fully connected DNN.
This architecture allows better representation of each data group and results in better
performance if the depths of the networks are properly selected. The output of GPN includes
eight design parameters, corresponding to the length, width, orientation, and existence of the five
elements (four arms and one connecting bar) of a general “H”-shaped particle. The second part
works on top of the GPN and functions as a spectrum-predicting-network (SPN), which receives
the predicted design parameters, material properties as well as a polarization indicator as an input
and returns the predicted transmission spectra as the outputs. Due to the twophase structure of
the network, backpropagation is optimized between the GPN and SPN for stability and
efficiency. With a training dataset containing over 16500 geometries simulated by an FEM
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solver, the desired performance was achieved. Note that although the generation of training data
is still a time-consuming process, the training takes only ~2 hours to get the best results. More
importantly, these efforts are nonrecurrent. Once the training is complete, a query to the DNN
about either the design (for a pair of given spectra) or the spectrum prediction (for a given
geometry) can be solved in a few milliseconds. In contrast, the same query to an evolutionary
algorithm or other traditional optimization methods would take much longer time to search the
entire parameter space. Figure 8(B) shows the representative results for two testing samples,
which were fabricated, measured, and composed of geometries not used in training. Excellent
agreement was achieved between the retrieved parameters and real dimensions measured by
scanning electron microscopy (SEM), and the spectra from measurements, predictions, and
simulations based on retrieved geometries also show fairly good overlaps.
The above framework optimizes a few parameters that describe a certain form of
geometries. While it shows the potential as a powerful tool of inverse design, in many
circumstances varying the geometry within a single class of topology is insufficient to generate
the intended complex optical responses. Liu et al. proposed an alternative method to explore the
enormous design space by employing a generative model [22], as shown in Figure 8(C). The full
architecture is constituted by three parts, namely the generator, the simulator, and the critic, all
being CNNs. Specifically, the generator and critic together function as a generative adversarial
network (GAN), which, unlike the previous case relying on paired input-output training data, is
an unsupervised learning system [139]. In practice, the simulator was pre-trained with 6500 full-
wave FEM simulations for a broad variety of shapes of gold nanoparticles. After training, its
weights were fixed, and the transmittance spectra of any input patterns would be approximated
by the simulator instead of being computed by full-wave simulations. The function of the
generator is to create unit cell patterns of the metasurface for input spectra T such that when the
generated patterns are fed to the simulator, the approximated spectra Tꞌ would largely replicate
the original inputs T. However, if there is no constraint on the training, the generated patterns
can be arbitrary and include numerous unrealistic results. On the other hand, if the true patterns
corresponding to T are directly used to determine the cost function, the model reduces to the
supervised case and the pattern generation becomes a deterministic problem. Therefore, the critic
plays a key role in the GAN. In the training process, it receives as inputs both the original
patterns resulting in T and the generated patterns in the form of images.
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The critic compares the two sets of images and restricts the generator to create patterns
that share common features with the original structures but are not identical to them. Figures
8(D) and 8(E) summarize two examples to show the performance of the network, which was
trained with several classes of geometries, such as circles, arcs, ellipses, crosses, handwritten
digits, and so forth. In Figure 8(D), the network responded correctly to the query of replicating
the transmittance spectra of an elliptical nanoparticle array. The generated pattern and resultant
spectra (right panel) only exhibit slight deviations compared with the inputs (left panel). This
retrieval can be achieved not only when the critic was fed with a single class of geometry but
also for a mixed training dataset that contains all the classes, meaning that the GAN can identify
the correct topology and conduct inverse design effectively. In Figure 8(E), after the critic was
trained with an incomplete set of handwritten digits, the network was asked to replicate the
spectra of a metasurface consisting of the missing digit “5”. Very interestingly, the generated
pattern was a modified “3”, which departs from the ground-truth but also contains some similar
geometric properties to reproduce most of the spectral features.
The fact that DNNs can learn complex functions from abstract data representations
provides unprecedented opportunities to the nanophotonic inverse design. In many applications,
the relations between desired functionalities and the design parameters are very intricate, and
physical insights and intuitive reasoning may not help to guide the design process. One example
is the generation of optical chiral fields. Chirality is a structural property of objects. An object is
chiral if it cannot be superimposed to its mirror image. Due to its universal existence in nature,
chirality has aroused enormous research interests [140-144]. From a nanophotonic point of view,
designing chiral nanostructures that respond to chiral light (usually left/right circularly polarized
(LCP/RCP) light) differently is of both fundamental and application-wise importance. However,
due to the complex, unrevealed underlying mechanism, despite some requirements on symmetry
being formulated to judge whether a structure is chiral or not [145,146], there is no general
guidelines that can be referred to if one wants to design a structure for a given chiral response or
how this response will evolve when the structure transforms. Taking advantage of deep learning,
Ma and coworkers demonstrated a purpose-designed learning architecture for implementing on-
demand design of three-dimensional (3D) chiral metamaterials [21]. Figure 9(A) illustrates the
general form of the unit cell, which consists of two layers of gold SRRs atop an optically thick
gold reflector. The two SRRs are sized l1 and l2 respectively and are twisted for an angle α.
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These parameters together with the spacer thicknesses t1 and t2 define the structure of the
chiral metamaterial. Upon illumination of LCP and RCP light, the metamaterial absorbs (or
equivalently, reflects) the incidence differently, resulting in circular dichroism (CD) signals. Like
in the previous cases, training a DNN to establish nonlinear mappings between the design
parameters and spectra is possible. And indeed, when the reflection spectra were fed into a
bidirectional DNN, denoted as the primary network (PN) in Figure 9(B), both spectra prediction
and design retrieval can be achieved after learning from 25000 simulated samples. Note that
compared with the fully connected DNN used in Ref. [20], here a tensor module is introduced to
account for the size mismatch between the low dimensional input vector of five design
parameters and the high dimensional output of spectra [147]. In Figure 9(C) (top panel), the
dashed and solid curves show the simulated and predicted spectra respectively. However, while
the two sets of spectra coincide well over most of the frequency range, obvious discrepancy
occurs at the steep resonance valley near 60 THz for the co-polarization reflection of RCP. The
reason of this degradation is that for each neuron in the output layer, the probability distribution
generated by the nonlinear function is centered at its off-resonance value. Sharp resonance
features deviating from the mean value is thus difficult to predict with high accuracy. A feasible
way to fix this issue is to combine the PN with an auxiliary network (AN) that associates the
design parameters directly to the CD signal (Figures 9(B) and 9(C), lower panels). Because CD
is not independent of the reflections, including CD spectrum in the dataset of PN will not
improve the leaning performance. When the predicted CD exceeds a threshold value, the AN
triggers a fine-tuning network in the combiner to locally refine the reflection spectra near that
frequency. The AN-corrected reflections are denoted by dotted lines in Figure 9(C), showing
excellent agreement with the simulated results.
This combined network enables on-demand inverse design of 3D chiral metamaterials
and discovery of some interesting, counterintuitive phenomena. For instance, in Figure 9(D),
chiral metamaterials with 10 and 170º twisting angles exhibit strong chiral responses at 60 THz
when the SRRs are in proper sizes; while if following the previous argument of symmetry, they
are close to the achiral structures where α = 0 or 180º and are not expected to generate strong
CD.
Another example of employing DNNs for inverse design without intuitive guidelines is
reported by Pilozzi et al. In Ref. [148], they applied a deep learning algorithm to solve the
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inverse problem for topological photonics. Stemming from the photonic analogue of quantum
anomalous Hall Effect in electronics, topological photonics studies the creation of interfacial
phonon transport or edge states that are protected from scattering [122]. The realization of such
systems with nontrivial topological properties usually requires using magnetism, time-domain
modulations or optical bianisotropy, but none of them can be easily designed for an intended
edge state at a given frequency, even in the simplest one-dimensional (1D) systems. Figure
10(A) illustrates the dielectric function profile of a multilayer structure with Harper modulation
[149,150], from which synthetic magnetic fields can occur. The existence of edge states and their
dispersion relations can be determined by assigning proper boundary conditions and solving the
eigenvectors of the transfer matrix.
The band diagram for a chosen modulation profile is shown in Figure 10(B), where the
green and orange strips correspond to the bandgaps, and edge states, indicated by the white
crosses, exist only in the gaps where a complex function Q changes sign. Provided a modulation
frequency and materials A and B, the search for layer thickness ξ and the phase χ of Harper
modulation for an edge state at frequency ωt cannot be solved analytically but can be addressed
by two DNNs. Because of the folding and multivalued branches of Brillouin zone, additional
categorical features were included in the dataset to label the different modes and different trends
of the iso-frequency surfaces. Unphysical solutions to the inverse problem were ruled out by
making a self-consistency check between the predicted frequency and the ground truth in the
training dataset. Figures 10(C) and 10(D) report the solutions from the direct and inverse DNNs
respectively, showing good agreement with the training set (colored curves).
A keystone of inverse design via deep learning, as a data-driven method, is the
sufficiently large training dataset, which is usually obtained by numerical simulations. For most
of the applications above, thousands of simulations are conducted for weeks to give a good
representation of the input space. To this extent, applications and geometries that are consistent
with analytical methods provides a suitable playground for efficient data generation. Figures
11(A) and 11(B) exemplify this by considering the scattering problem of a SiO2/TiO2 multilayer
spherical particle [151]. With analytical solutions derived from the transfer matrix method,
50000 samples were generated for different combinations of shell thickness. A fully connected
DNN was used to solve the inverse design task. As can be seen in Figure 11(A), for an arbitrarily
chosen sample spectrum from the test set (blue curve), the DNN successfully captures all the
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spectral features with only moderate deviations in amplitude at a few peaks/valleys (red dotted
curve). In contrast, when a nonlinear optimization based on the interior-point method was
employed to solve the same problem, much larger inconsistency is observed (black dashed
curve). In fact, as the number of design parameters increases, optimization methods tend to
become stuck in the local minima instead of the global ones, while DNNs are not affected.
Moreover, DNNs can be easily adapted to fit different design requirements. For example,
by using a different cost function, a SiO2/silver multilayer particle showing broadband scattering
within the desired wavelength range was found from the enormous possible candidates (Figure
11(B)). The similar procedure has also been used to study the transmission of multilayer thin
films, where hundreds of thousands of samples were generated for training [152]. Since for such
non-resonant structures there is a high likelihood that different configurations can result in nearly
identical optical responses, a tandem network was proposed to overcome the non-uniqueness
issue. As shown in Figure 11(C), a pre-trained network solving the direct problem was connected
via an intermediate layer M to the original DNN. This architecture works in a similar way to the
bidirectional networks, which applies additional constraints to the learning process. The
modified network gives reasonable designs even when asked to fit some unrealistic spectra as in
Figure 11(D). More complex functions, such as achieving phase delays at multiple wavelengths,
can be realized by using structured thin films (Figure 11(E)) and modifying the network structure
accordingly, while generating new data set is not a high cost.
Another aspect of the considerations about data is the volume. So far, the applications of
deep learning in nanophotonics are limited to establishing the mappings between the design
parameters and the optical responses given by spectra. The distributions of electric and magnetic
fields and their derived quantities in the 2D or 3D space are also of great interest. However,
using 2D or 3D field distributions as data sets is not practical. On one hand, this leads to huge
amounts of data that are unaffordable for storage and training, especially in nanophotonics where
ensuring high spatial resolutions is a basic need. On the other hand, how the feature
representation in 2D and 3D data can be effectively utilized is largely unexplored. Barth and
Becker proposed an interesting technique based on a machine learning algorithm, though not
deep learning, for the classification of the photonic modes in a PhC [153] (Figure 12(A)). This
task is aimed at a type of applications different from inverse design. Taking sensing for example,
the 3D field distributions associated with nanostructures need to be evaluated by some criteria
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and optimized accordingly in order to facilitate the light-molecule interactions and maximize the
performance. However, the analysis of field distributions is difficult, usually solved by
visualizations and processing the full set of 3D data.
Taking advantage of an algorithm based on Gaussian mixture method (GMM) [154], Ref.
[153] showcased that the clustering model can reduce the 3D field distributions to a finite
number of distribution prototypes, which allows the identification of characteristic photonic
modes. Figure 12(B) compares the band diagrams of a PhC obtained by two methods: The left
panel is calculated by an integration of electric field energy density over the volume within the
hole and a thin layer above the PhC, while the right panel depicts the classification map of field
distributions on the three symmetry planes marked in Figure 12(A). A fairly good match is
observed, which confirms the validity of the procedure. Moreover, based on the clustering
results, the field distribution prototypes can be obtained. As shown in Figure 12(C), by
inspecting the field distribution of each cluster, leaky modes that result in strong near-fields can
be distinguished from the radiative modes. These results were further validated by finite element
simulations in Figure 12(D), where the illumination conditions were determined using the
silhouette coefficients for classification [155]. Therefore, with a lower dimensional dataset, the
proposed technique provides an alternative approach to extracting information from 3D field
distributions that may not be accessible via visualization-based analysis.
4. DEEP LEARNING ON NANOPHOTONIC
Platforms The arrival of the era of big data has rendered the speed, energy consumption,
and information density of computing the key considerations in hardware development.
However, after decades of continuous improvements following Moore’s law, electronics started
facing bottlenecks on these aspects, which are physically fundamental and can no longer be
resolved by scaling. Integrated photonic circuits are considered promising candidates to
overcome the above obstacles, because of the higher speed and energy efficiency associated with
photons. On the other hand, traditional electronic components such as the central processing
units (CPUs) are not well suited to serve the emerging techniques in artificial intelligence. New
hardware architectures aimed at accelerating AI and deep learning are also in a pressing need.
Within the domain of electronics, GPUs, vision processing units (VPUs), tensor processing units
(TPUs) [156], TrueNorth [157] and other integrated chips [158] have been developed and
showed great potential in practical applications. Meanwhile, hybrid opto-electronic systems for
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implementing spike processing [159,160], neuromorphic computing, and reservoir computing

have also been demonstrated, and progress is being made towards their photonic realization.
Some timely reviews have given comprehensive discussions on these topics [26,28,30]. In this
section we keep our focus on the all-optical implementation of DNNs.
While photonic circuits in general operate at higher speeds and with higher energy
efficiencies compared with their electronic counterparts, implementing DNNs and computing on
photonic platforms offers a few advantages in this specific application [23,161,162]. First, as
discussed in the previous section, the computation of DNNs is mostly achieved by matrix
multiplications. In nanophotonic circuits, linear matrix operations can be performed very fast—
almost at the speed of light, and, in parallel and efficiently due to the non-interacting nature of
photons. Second, the nonlinear functions in the DNNs can be realized by optical nonlinearities in
photonic circuits, such as saturable absorbers or amplifiers. Third, for a given photonic DNN,
after training, the whole system is passive and consumes no power. Last, it is possible to conduct
training of photonic DNNs by an optical means. This could significantly accelerate the learning
process and further reduce power consumption.
One of the earliest demonstrations of photonic DNNs was reported by Shen et al. [23],
where vowel recognition was achieved showing comparable performance to a 64-bit electronic
computer. To implement the full map of a DNN after training, each layer of the network is
composed of an optical interference unit (OIU) to carry out the linear matrix multiplication and
an optical nonlinearity unit (ONU) that acts as the nonlinear activation. Four different vowel
phonemes spoken by 90 different people were used to train and test the circuit, which were first
preprocessed on a computer and then fed into the nanophotonic DNN as amplitude-encoded
optical pulses to generate outputs, as shown in Figure 13(A). The physical realization of the
network in a nanophotonic circuit is not as straightforward as it appears in the schematic.
Because after training the weights may end up with an arbitrary distribution, the design of OIUs
needs to tackle the problem of how the propagation of optical pulses through the unit can be
equivalent to a multiplication by an arbitrary matrix. Fortunately, a real-valued matrix M can be
expressed by M = U∑V† via singular value decomposition [163], with U and V † denoting two
unitary matrices and ∑ a diagonal matrix. In nanophotonic circuits, a unitary matrix can be
implemented with beam splitters and phase shifters [161,164], and a diagonal matrix can be
realized by using optical attenuators or amplifiers [165,166].
ISBN: 978-81-948129-1-3 Page 395
Therefore, a proper arrangement of these optical components is capable of performing

matrix multiplications. Figure 13(B) shows the optical micrograph of an OIU fabricated on a
programmable nanophotonic processor. The unit consists of 56 Mach-Zehnder interferometers
(MZIs) with each of them containing two phase shifters and a directional coupler to achieve
desired functionalities via programming. The red- and blue-highlighted meshes denote the
components that perform the unitary and diagonal matrix multiplications, respectively. Input
optical pulses propagate through the unit, producing the correct interference patterns at the
output. A similar scheme was adopted for implementing a different processor architecture for
quantum transport simulations [167]. In Ref. [23], instead of physical realization, the nonlinear
activation was simulated on a computer as saturable absorbers. Whethher real nonlinear optical
elements can work equally well is still an open question to be addressed. Figure 13(C) compares
the correlation matrices of a bilayer nanophotonic DNN and a computer for the vowel
recognition task. The nanophotonic DNN shows a correctness of 76.7%, lower than but still
comparable to the result of 91.7% from the electronic processor. Larger error from the
nanophotonic prototype turns out to come from the photodetection noise, limited phase encoding
resolution, and thermal crosstalk between phase shifters. Nevertheless, all these error sources can
be relieved by different strategies in future implementations.
The above nanophotonic DNN still relies on regular computers at the training phase. This
makes the whole process inefficient. An on-chip training scheme based on forward inference was
proposed, which in principle could fit complex network architectures such as CNNs and
recurrent neural networks (RNNs) where the effective number of parameters is substantially
more than the distinct number of parameters. However, conducting such training requires
repeatedly tuning every MZI in the circuit and is not efficient when the chip is scaled up. Hughes
et al. developed an alternative protocol in which the on-chip training is accomplished only by in
situ intensity measurements [168]. When a DNN is implemented by a nanophotonic platform
employing the architecture in Ref. [23], the gradient terms of the cost function computed from
backpropagation physically correspond to the error derivatives with respect to the permittivity of
the phase shifters in the OIUs.
Interestingly, this gradient distribution can be expressed as the solution to an
electromagnetic adjoint problem. With the assistance of the adjoint variable method (AVM), an
optimization technique used in the inverse design of photonics [169,170], the gradient at a phase
ISBN: 978-81-948129-1-3 Page 396
shifter is given by the overlap of the optical fields from the “original” and “adjoint” problems.
Figure 13(D) summarizes the training procedure demonstrated by simulations. The same flow
applies if a real circuit is fabricated for experiments. The circuit has 3 input ports and 3 output
ports to perform a 3×3 unitary matrix multiplication (panel (i)). Training starts by sending an
(original) input vector X from the left, e.g. [0 0 1]T as in panel (ii). The intensities of the light
field at each phase shifter is measured and stored as Iog. Next, an adjoint field δ, e.g. [0 1 0]T in
panel (iii), is fed into the circuit from the output ports on the right. The field intensities are also
measured and stored as Iaj. Based on the resulting field pattern, the time-reversed adjoint field
XTR is calculated, which is then added to the original input X to feed the unit from the input
ports, resulting an interference pattern in panel (iv) and intensities I at each phase shifter. The
final computation of the gradient is done simply by subtracting Iog and Iaj from I, followed by a
multiplication by a constant.
The bottom two panels show the comparison of the gradient information, which are
obtained by AVM with simultaneous excitations at both sides (panel (v)) and by the optical
method (panel (vi)), namely interfering the patterns in panels (ii) and (iv), respectively. The very
good agreement confirms that the gradient terms can be determined by in situ intensity
measurements at the phase shifters. This result is significant, because it allows the computation
in parallel of the crucial gradient distribution. With two OIUs in Figure 13(D) connected in
series, a logic XOR gate was demonstrated. Figure 13(E) reports the network predictions before
and after training, where the latter shows obvious improvement, matching perfectly with the
truth table.
The above integrated nanophotonic neural networks are composed of hundreds of
components to implement their functionalities. When the platform is scaled up to involve
thousands or even millions of artificial neurons, the complexity of the achievable tasks can be
dramatically boosted. Taking advantage of the analogy between information transformation
through DNNs and light diffraction in layered structures, in a recent work, Lin et al.
experimentally demonstrated various complex functions with an all-optical diffractive deep
neural network (D2NN) [171]. The mechanism is illustrated in Figure 14(A). Recalling the
diagram in Figure 7, in a DNN, data are transformed from neuron to neuron via their
interconnections. In a layered diffractive structure, according to the Huygens-Fresnel principle,
each single point on a certain layer acts as a secondary light source.
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The individual points are excited by the incoming light waves from points in the
preceding layer and emit light to the subsequent layer, resulting in, in theory, full “connectivity”
between adjacent layers via diffraction and interference. Despite similarities in the layered
structure and ways of connection, there are several differences between D2NNs and conventional
DNNs. In standard DNNs, real-valued weights are associated with neuron connections, the
neurons carry a nonlinear activation and an additive bias term, and the output of each neuron is
the weighted sum of the real-valued inputs, computed by matrix multiplications. In contrast,
D2NNs are complex-valued due to the nature of the optical waves. The output of each point is
given by the product of the input wave and the complex-valued transmission or reflection
coefficient at that point. In this process, the local transmission or reflection coefficient applies a
multiplicative bias to the output wave at that point, which is then weighted through propagation
to interfere with other secondary waves at points on the next layer, physically implementing the
matrix multiplications.
Two functions, namely a classifier (Figure 14(B)) and an imaging lens (Figure 14(C)),
were experimentally demonstrated at 0.4 THz using 3D-printed D2NNs with phase-only
modulation, while the training process was still completed on a computer. The complexity of the
tasks significantly increases the required number of neurons and of the training data. For the digit
classifier, 55000 images of handwritten digits (0-9) were used to train a five-layer D2NN
composed of ~0.2 million neurons. Because of their more abstract feature representation than
digits, fashion products could be daunting for classification. But remarkably, the numerical tests
of a five-layer network showed a classification accuracy of 81.13%, and the experiments reached
a 90% match with this result. Figure 14(D) shows a representative example of a sandal input
image and the corresponding output intensity map (left column). As expected, most energy is
directed to detector 5. The confusion matrix and energy distribution for the whole experimental
test dataset can be found in the right column. Although the reported classification accuracy is
lower than the record of 96.7% from state-of-the-art CNN algorithms [172], this D 2NN uses
almost one order of magnitude fewer neurons and its performance can be improved by
introducing amplitude modulation, additional layers, and possibly optical nonlinearity.
Lastly, DNNs are sometimes described as “black boxes”, because it is almost impossible
to extract an intuitive picture to explain how data are processed through the hidden layers.
D2NNs may provide some insights into this. In Figure 14(E), three spatially separated Dirac-
ISBN: 978-81-948129-1-3 Page 398
delta functions are fed into a D2NN-based unit-magnification imager composed of 10 layers of
phasemodulation masks (left panels). It can be seen from the amplitude and phase distributions
that indeed each neuron is connected to various neurons in the next layer in an abstract way. The
input neither propagates as needle-sharp beams nor diffracts as in the free-space (right panel).
Rather, each delta function tends to be diffused at the beginning, attenuated in the halfway, and
finally focused to the same point of the output plane as where it is emitted on the input plane.
Visualizing wave propagation through the D2NN for this specific application help to reveal the
operation principle of the coherent optical DNNs.
5. CONCLUSIONS AND OUTLOOK
In this review, we have summarized the recent advances on nanophotonics that are
enabled or powered by advanced computational methods, especially deep learning algorithms. In
the inverse design of nanophotonic devices, these techniques allow us to go beyond physical
insights and help to search the parameter space in a more efficient way, leading to data-driven,
on-demand design of novel devices. In the opposite direction, the development of nanophotonics
could provide new platforms that can potentially overcome the bottleneck in computing power
for machine learning. As the research interests and efforts on this topic continue increasing, we
envisage that the following directions will be promising in the next stage of development.
First, advanced optimization techniques, especially gradient-based topology optimization that
can handle up to 1 billion design variables efficiently [73,173], allow for the design of
nanophotonic devices with tremendous complexity. Whereas this capability has been applied to
the aerospace industry, the design resolution for nanophotonics is much lower. Increasing the
number of design variables will empower the invention of more sophisticated and more
integrated metadevices.
Second, the current application of deep learning in nanophotonic inverse design is still
limited to finding appropriate design parameters for the desired spectra. Using low-dimensional
inputs such as spectra and diffraction efficiencies [174] in the network ensures the data volume
will not diverge but also restricts the achievable functionalities. Higher dimensional data such as
the field intensity profiles as well as vectorial field maps carry much more information that can
be used for designing functional metalenses and holograms. Blooming of nanophotonic devices
enabled by deep learning is expected once the difficulties in computation power and data storage
are overcome.
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Third, the mechanism of many optical effects and multiphysics processes involving
optics has not been well understood, which hinders the physics-inspired design but is where
machine learning can come into play. For instance, although DNNs can provide accurate
solutions when an arbitrary CD spectrum is desired, engineering the near-field optical chirality
arising from the complex interplay between electric and magnetic fields is a task far from being
solved. Similarly, optimizing optical forces at the nanoscale is critical for optical tweezers [114-
117,175] and sorting [176-178]; nonlinearities of nanostructures for efficient harmonic
generation and optical switching also have plenty of room to improve towards functional
circuitry [179]. The lack of design guidelines makes them suitable problems for data-driven
methods to deal with.
Next, despite the recent success in implementing DNNs on nanophotonic circuits and
THz platforms, the all-optical realization of DNNs has not been fully demonstrated. For
example, the training process is mostly conducted on electronic chips or computers, which does
not really fulfill the advantages over speed and energy consumption. On-chip training, as
numerically demonstrated, can overcome this limitation, while whether the losses will diminish
the performance in a real system, and, how the measurement-based training can be effectively
performed in large scale networks containing at least thousands of neurons, are potential issues
to be addressed. D2NNs offer an alternative mechanism and platform to photonic circuits,
whereas scaling down the THz scheme to visible or telecom wavelengths is demanding due to
the limited fabrication resolution, interparticle coupling, and material losses. In addition,
although demonstrated elsewhere, optical nonlinearity has not been introduced in photonic
DNNs. Therefore, resolving these challenges will be essential steps towards all-optical DNNs
[180].
Lastly, the use of machine learning techniques in nanophotonics has just emerged.
Among the early attempts introduced in this review, many of them use standard network models,
which may not be the best fit for the target applications. It is possible that the demonstrated
performance can be simply improved by reforming the feeding data or modifying the network
structure. Recurrent neural networks (RNNs) that can learn from sequential inputs have been
realized on a photonic platform very recently but not yet in nanophotonics [181,182]. Including
time domain features in the dataset will be very attractive. Furthermore, other leaning paradigms,
such as unsupervised learning and reinforcement learning, and combinations of deep learning
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and other computational methods are expected to provide new design frameworks that are faster,
more accurate, and even independent of human knowledge [183].
Nanophotonics and machine learning are two research domains that differ from the very
basis. While it is promising to apply machine learning methods to data-driven nanophotonic
design and discovery, many of the techniques, mature or cutting-edge, are not well known by the
photonics community. Therefore, bridging this knowledge gap is pressing. Significant
advancement will come out with further combination of the two fields.
Figure 1.A dial illustration of computational methods and their potential applications in
nanophotonics. Items on the circumference indicate different applications (not in the order of
timeline or importance), where computational methods can be employed in the design process.
The second hand corresponds to traditional schemes based on numerical simulations. Many
cycles of trial-and-error modeling are needed for a specific task, each giving an incremental
advancement towards the final goal. The minute hand denotes various optimization techniques,
such as genetic/revolutionary algorithms and gradient-based approaches. Supported by
simulation techniques, optimization methods search the full parameter space for each task by
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minimizing the cost function, providing a more efficient framework for achieving complex
functionalities. The hour hand represents deep learning. Although a sufficiently large amount of
data needs to be generated (by simulations) first for training, once the training is complete, the
network solves a design request almost instantaneously
Figure 2.Demonstrations of on-chip devices based on inverse design. (A) A photonic crystal
waveguide Z-bend showing exceptional transmission. Panels in zigzag order: Schematic of
optimization; simulated wave propagation; SEM image of the fabricated Z-bend; comparison of
bend losses between optimized (thick grey/red for measurement/simulation) and unoptimized
(thin black) structures. (B) A two-channel wavelength splitter. Left: SEM image. Middle:
Simulated field patterns for 1300 (blue) and 1550 nm (red) wavelengths. Fields are superimposed
and color-coded for illustration. Right: Measured transmission spectra. (C) A nanophotonic
mode-converting polarization beam splitter. Top: SEM image and comparison of measured (solid
lines) and simulated (dashed lines) transmission of the fabricated device. Bottom: Simulated
field intensity distributions for TE and TM modes at 1550 nm. (D) Simulated magnetic field
distributions in an optical diode when the excitation is on the left (left panel) and right (right
panel), respectively. (E) SEM image of a photonic reflector, as part of an on-chip Fabry-Pérot
cavity. (A) is reprinted with permission from Ref. [74], OSA; (B) is reprinted from Refs. [11]
ISBN: 978-81-948129-1-3 Page 402

and [76] by permission from Springer Nature; (C) is adapted from Ref. [80] by permission from
Springer Nature; (D) is reprinted from Ref. [71] with permission (CC BY 4.0); (E) is reprinted
from [82] with permission.
Figure 3.Metasurface inverse design using topology optimization. (A) A five-wavelength beam
splitter with the SEM image (left), experimental (middle) and theoretical (right) diffraction
efficiencies. Inset: Correlation between incidence wavelengths and deflection angles. (B) Design
of a multilayer focusing metalens with angular aberration correction. Rectangles in black denote
silicon resonators, and the grey background is alumina. (C) Simulated far-field intensity profiles
for the structure in (B) at four angles of incidence follow the identical diffraction limit.
Intensities are normalized to unity for comparison. (A) is reprinted with permission from Ref.
[83], Copyright 2018 John Wiley and Sons; (B) and (C) are adapted with permission from Ref.
[89], Copyright 2018 American Physical Society.
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Figure 4.Nanophotonic devices based on evolutionary design and optimization. (A) Illustration
of procedures of evolutionary design. (B) The SEM image (left) of a metasurface that can
generate five focal points (right) arranged in a T-shape. (C) A simplified model of optimized
matrix nanoantennas for improving the near-field intensity enhancement. (D) SEM images of
optimized silicon nanoantennas for polarization-encoded color display. The associated
wavelengths denote the resonances for y-polarized incidence, while for x-polarization, the
resonance is targeted at 550 nm. (E) Measured scattering spectra for x- (top) and y-polarized
(bottom) incidence. (F) Optimization history of the averaged absorption enhancement in a quasi-
random light-trapping structure. Insets show the generated patterns at selected optimization
steps. (A) and (B) are reprinted from Ref. [90] with permission; (C) is reprinted from Ref. [92]
with permission, Copyright 2018 American Physical Society; (D) and (E) are adapted from Ref.
[94] by permission from Springer Nature; (F) is reprinted from Ref. [95] with permission,
Copyright 2018 National Academy of Sciences.
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Figure 5.Topology-optimized cloaks and concentrators. (A) A 2D low-contrast all-dielectric

cloak. Left: The dielectric layout for four symmetry lines. Inner circle denotes an ideal metallic
cylinder. Middle: Numerical demonstration of the cloaking performance. Right: The scattering
pattern of a bare cylinder. (B) A 2D unidirectional cloak with a low refractive index material
(blue region, εr = 2) and the simulated scattering pattern. (C) Experimental realization of the
design in (B) at microwaves. (D) Design of a 3D magnetic field concentrator (left) and the
simulated magnetic field distribution (right). (A) and (B) are adapted from Refs. [106] and [107],
respectively, with permission of AIP; (C) is reprinted from Ref. [108] with permission of AIP;
(D) is reprinted from Ref. [110] with permission.
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Figure 6. (A) Inverse design of illumination patterns to maximize optical torques. (B) Optimized
torque spectra and field distributions for three target wavelengths at 1028, 805, and 625 nm,
respectively. (C) Inverse design of PhCs for enhancing spontaneous emission. Left: Dielectric
layout of the design. Black regions correspond to a material with refractive index n = 2. Middle:
LDOS profile at the third-order Dirac exceptional point. Right: LDOS spectra at the center of the
unit cell. (D) A binary plasmonic structure (top) designed by simulated annealing algorithms to
produce five focal points of SPPs. (E) A super-oscillatory lens designed by particle swarm
optimization for subwavelength imaging. Top: SEM images of the fabricated lens (left) and a
cluster of nanoholes in a metal film (right). Bottom: Images of the cluster by a conventional lens
(left) and by the super-oscillatory lens (right). (A) and (B) are reprinted with permission from
Ref. [120], OSA; (C) is adapted with permission from Ref. [121], Copyright 2018 American
Physical Society; (D) is adapted with permission from Ref. [123], OSA; (E) is adapted from Ref.
[124] by permission from Springer Nature.
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Figure 7. The architecture and learning process of a deep neural network comprising multiple
layers. The circles represent artificial neurons. In the shown case, neurons in the same layer do
not interact, but each neuron is connected, with a unique weight wij, to every neuron in the
adjacent layer(s), namely the preceding and/or subsequent layers. Data transformation between
adjacent layers is implemented by linear matrix multiplications of the weights and input vectors,
followed by the application of a nonlinear function f (and sometimes an additive bias bj) at each
neuron. Left: Forward inference in which data flow through the network from the input layer to
the output layer. Right: Training with backpropagation, where every weight value is adjusted
based on the error derivative to minimize the cost function. The forward inference,
backpropagation, and weight update are repeatedly performed as data are continuously supplied,
until the desired performance is obtained.
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Figure 8. (A) A DNN for design and characterization of metasurfaces. The network comprises a
layered GPN (left) to solve the inverse design problem and an SPN (right) to predict the spectra
based on retrieved design parameters. (B) Demonstration of design retrieval and spectra
prediction based on (A). The building block is a gold nanostructure with its geometry
represented by a general “H” form. (C) Architecture of a generative network composed of a
generator, a pre-trained simulator, and a critic. In the training phase, by receiving spectra T and
the corresponding patterns X respectively, the generator and the critic learn jointly. Valid
patterns are smoothened to binary maps and stored as candidates for metasurface design. (D)
Original (left) and generated pattern (right) and their transmittance spectra based on a training
dataset of elliptical nanoparticles. (E) Original (left) and retrieved pattern (right) and their
transmittance spectra based on an incomplete dataset of handwritten digits. The network
generated a modified “3” to best replicate the spectra for pattern “5”, which was removed
intentionally from the training data. (A) and (B) are reprinted from Ref. [20] with permission(CC
ISBN: 978-81-948129-1-3 Page 408

BY 4.0); (C)-(E) are reprinted with permission from Ref. [22], Copyright 2018 American
Chemical Society.
Figure 9. (A) The unit cell of a chiral metamaterial consisting of two layers of gold SRRs on top
of a reflective mirror. Two SRRs are twisted by an angle α, which breaks the mirror symmetry of
the system and determines the chiral response together with other design parameters, including
the sizes of SRRs and the thicknesses of the spacing layers. (B) A combined architecture
consisting of two bidirectional DNNs. The primary network (top) connects design parameters
and reflection spectra, and the auxiliary network (bottom) creates mappings between design
parameters and CD with higher accuracy. Data are allowed to flow between PN and AN via
combiners to refine resonance features in all the spectra. (C) Top: Comparison of reflection
spectra obtained by simulations (dashed lines), by PN alone (solid lines), and by the combined
system in (B) (dotted lines). Bottom: Comparison of simulated CD and AN predicted CD. (D)
Evolution of CD at 60 THz for different combinations of SRR sizes and selected twisting angles.
Figures are adapted with permission from Ref. [21]. Copyright 2018 American Chemical
Society.
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Figure 10. (A) Dielectric function profile of a multiplayer structure with Harper modulation.
Layers with material A are stacked with spatial modulation along the z-axis in a homogeneous
bulk material B. (B) The band diagram of the structure in (A). Orange and green ribbons
represent bandgaps. Edge states are denoted by regions with crosses, which exist only in the
bandgaps where a complex quantity Q changes sign. White and purple regions correspond to Q >
0 and Q < 0, respectively. (C) The band diagram predicted by the direct DNN. (D) The
modulation diagram retrieved by the inverse DNN. In (C) and (D), colored curves represent the
training dataset. Figures are adapted from Ref. [148] with permission (CC BY 4.0).
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Figure 11.Inverse design of multilayer structures via deep learning. (A) A DNN retrieves the
layer thicknesses of a multilayer particle based on its scattering spectrum, showing much higher
accuracy than the nonlinear optimization method. The main figure compares the scattering
spectra by simulation (blue), optimization (black), and prediction of DNN (red). Comparison
between the ground truth and retrieved design parameters are given in the legend. (B) Inverse
design of multilayer particles for broadband scattering in a given wavelength range. (C) A
tandem network for designing multilayer thin films. A pretrained forward modeling network is
connected to the inverse design network to avoid non-uniqueness issues. (D) Performance of the
tandem network in fitting a Gaussian profile in the frequency domain. (E) Structured multiplayer
thin films for modulating the transmission phase delay. (A) and (B) are reprinted from Ref. [151]
with permission (CC BY-NC); (C)-(E) are reprinted with permission from Ref. [152], Copyright
2018 American Chemical Society.
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Figure 12. (A) The unit cell of a silicon PhC on glass, in which circular holes form a hexagonal
lattice. When illuminated by external light from the top, leaky modes are excited, which exhibit
strong near-fields boosting the emission of the nearby emitters, such as quantum dots. Rectangles
in color denote the symmetry planes used for exporting field data. (B) Comparison of the
simulated band diagram (left) and clustering results (right) when the sample is oriented in the Γ-
K configuration and irradiated by a TEpolarized plane wave. Left: The diagram of volume-
averaged electric field energy enhancement. Right: The classification map depicted by blending
the color-coded silhouette coefficient of each mode with a black background. (C) Top view of
electric field energy plots for the eight prototypes found in the clustering, with three of them
associated with leaky modes exhibiting strong near-fields. Mode A: plateau mode; Mode B:
flank mode; Mode C: hole mode. Modes are termed base on the location of the field
enhancements. (D) 3D semi-artistic plots of the interactions between leaky modes and randomly
distributed quantum dots, which emit light with intensity proportional to local field energy
density. Figures are adapted from Ref. [153] with permission (CC BY 4.0).
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Figure 13.Nanophotonic DNNs. (A) The architecture of a nanophotonic neural network for
vowel recognition. Each box in grey corresponds to an optical interference unit that computes
matrix multiplication, followed by an optical nonlinearity unit connecting it to the next layer. (B)
Optical micrograph of the optical interference unit used in the experiments. The unit comprises
56 programmable MZIs, with the red/blue mesh highlighting the functional part implementing a
multiplication by a 4×4 unitary/diagonal matrix. Inset: Layout of the MZIs composed of two
phase shifters and a directional coupler. (C) Confusion matrix of the nanophotoniccircuit (left),
in comparison to that of a 64-bit computer (right), for vowel recognition by a network with two
hidden layers. The elements (X,Y) of the matrices are the numbers of times a spoken vowel X is
identified as Y. Perfect identification would give a diagonal matrix. (D) On-chip training of a
nanophotonic OIU through in situ measurements. (E) An optically trained nanophotonic DNN
implementing a logic XOR gate. The tables compare the network predictions before (left) and
after (right) training. Target answers (truth table) are depicted with crosses and predictions are
denoted by circles. (A)-(C) are adapted from Ref. [23] by permission from Springer Nature; (D)
and (E) are adapted with permission from Ref. [168], OSA.
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Figure 14. All-optical machine learning by D2NNs. (A) A D2NN consisting of multiple
transmissive layers. Each point (color coded squares) acts as a neuron by providing a
multiplicative complex-valued transmission coefficient to the incoming wave. The coefficient
distribution is trained by deep learning algorithms to perform a predefined function and is fixed
after fabrication. (B) Schematic of a 3D-prined N2DD implementing a classifier for handwritten
digits and fashion products. Different types of objects on the input plane leads to maximized
light intensity at the corresponding detector on the output plane. (C) Schematic of a 3D-printed
N2DD implementing a lens for imaging. The output is a unit-magnification image of the input
optical field. (D) Classification of fashion products. Left: An input image belonging to a certain
class of products, e.g. sandals, results in maximum energy at the corresponding detector on the
output plane. Right: Confusion matrix and energy distribution for the full test dataset of the
fashion products classifier. (E) Wave propagation in a N2DD imager with 10 layers. Left:
Amplitude and phase distributions for an input of three Dirac-delta functions passing through the
network. Right: Amplitude distribution for the same input passing through a vacuum. Diagrams
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at the bottom show the light intensities on the output plane. Figures are reprinted from Ref. [171]
with permission from AAAS.
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ISBN: 978-81-948129-1-3 Page 427

EVALUATION OF CHITOSAN-BASED BIOMATERIALS AS BIOMEDICAL

APPLICATIONS
K.Anu1 and Nitika Tomar2
1
Department of Zoology, Auxilium College (Automomous), Vellore-632007.
E.mail: anukrishnachander89@gmail.com
2
Department of Chemistry, Raja Balwant Singh Engineering Technical Campus, Bichpuri, Agra
Email: nitikakuss@gmail.com
ABSTRACT
Chitin, the second most available marine biopolymer, was firstly used in the 1930s and
early 1940s. It has a linear structure that is composed by repeating β-1,4 linked N-acetylated
glucosamine units. Chitosan, derived from chitin, is a natural polymer that is known for being
biodegradable, non-toxic and antibacterial. It is cationic, has a hydrophilic surface and causes
minimal foreign-body reaction. Also, chitosan shows a reasonable solubility in water and in
organic acids. However, this represents the major limitation to chitosan’s preparation and use for
biomedical applications. Due to these properties it can be used for drug delivery, cancer
diagnosis, immune regulation, tissue regeneration (treatment of damaged skin, nerve
regeneration, cartilage and bone repair) and as antioxidant and anti-inflammatory. Also, shows
an antitumor and antidiabetic capacity and haemostatic effects. In what Dental medicine is
concerned, chitosan is very useful too. Due to its bone regeneration capacity, it can be used as a
therapy for alveolar bone repair, as well as in the implantology field. Because of its antimicrobial
effect, is very promising in endodontic treatment. Also, it promotes a higher resistance to fungal
diseases and leads to better effectiveness on decay remineralization. In addition, although
numerous treatments are used to regenerate the alveolar bone, cementum, and loss of periodontal
ligaments (PDL), Chitosan-based therapies are used to treat periodontal lesions. To conclude,
chitosan is an innovative biomaterial which use has been increasing exponentially over the last
years.
KEYWORDS: chitosan; chitin; biomaterials; biomedical application; tissue regeneration; drug
delivery
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INTRODUCTION
The commercial applications of chitin arose in the 1930s and early 1940s but it was first
identified in 1884. [1, 2] Chitin consists of a marine biopolymer which is the second most
available in nature (the main one is cellulose) [2-4]. It has a linear structure that is composed by
repeating β-1,4 linked N- acetylated glucosamine units. [2, 3, 5, 6] Chitin can occur as α, β or γ
forms. Each one has its own structural properties:
• The α form has microfibril with an antiparallel orientation which are linked by strong hydrogen
bonds. In nature, it is the most common form and is the best one for the industry.
• The β form, unlike α, has its chains parallel orientated (linked by weak hydrogen bonds).
• The γ form, that is the less known, consists of a mixture of antiparallel and parallel chains.
Due to strong hydrogen bonds present in α- chitin, it is insoluble in water, most organic acids
and dilutes the acid and alkaline solutions. On the other hand, since β-chitin has weak hydrogen
bonds, it has better solubility in most acids and swells up in the water. [4]
This polymer can be found in cell walls of yeasts and fungi, shells of crustaceans, corals, diatoms
sponges, molluscs, the cuticles of insects and worms, however, the major commercial sources of
chitin are crab and shrimp shells. [1, 5, 6, 14, 25] Bearing this in mind, chitin extraction from
shells unable us to minimize the waste, as well as to produce compounds with valuable
biological properties and specialty applications, such as chitosan [1, 4].
MATERIALS AND METHODS
As a starting point, we used the terms “Chitosan collagen membrane”. As a result, we
obtained 174 articles. Posteriorly applying filters, such as “5 years” and “free full-text”, this
number reduced to 17, from which we chose 10 after reading their abstract. Once the number of
articles we obtained was insufficient to elaborate a review, we were forced to enlarge de research
and, consequently, the theme, looking for more embracing terms.
Therefore, in the second phase, we searched for "Chitosan". It resulted in 21608 articles, a
number that reduced to 63, after applying for the filters "review", "free full-text", "5 years" and
"humans". From those, 13 were found pertinent by Reading the abstract.
Secondly, “chitosan membrane” was the expression searched. 2759 articles were gathered,
resulting in 12 after using the filters “review”, “free full-text” and “5 years”. Due to reading the
abstract, the number of selected articles decreased to 5.
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In another research, we searched for “Chitosan membrane AND dentistry”. Ultimately,

“Chitosan scaffold AND dentistry" was the expression we looked for. 75 documents were
founded, however, only 2 were selected by reading the abstract. The filters used were "free full-
text" and "5 years". To conclude, some articles were common to the different researches that we
made. Those were only considered once.
Chitosan
Chitin is transformed into chitosan, its main derivative, by partial deacetylation under
special conditions like a strongly alkaline environment. Chitosan is a linear amino
polysaccharide composed of β(1→4) glycosidic bonds linking, randomly distributed, D-
glucosamine, and N-acetyl-D-glucosamine monomers. [5-8, 16, 25, 26] It is structurally similar
to the glycosaminoglycans in the extracellular matrix. [8, 11]
Figure 1- Chemical structure of chitin and chitosan
Deacetylation can be performed by enzymatic preparations or chemical process, being the last
one the most used because of its low cost and as it allows massive production. (1)
Chemical Deacetylation
Chemical deacetylation can be performed by acids or alkalis. However, as glycosidic
bonds are very susceptible to acid, alkalis (such as NaOH) are more often used. Depending on
the NaOH concentration, reaction time, temperature and repetition of alkaline steps, final
chitosan may have different characteristics, such as degree of deacetylation (DA) and molecular
weight (MW). Other factors may be taken [5] into consideration: atmosphere, reaction reagent,
chitin and solvent ratio, particle size and source of raw material. Despite its advantages, chemical
deacetylation has some cons like energy consumption and waste of concentrated alkaline
solution. As a result, enzymatic deacetylation has occurred. [1, 25]
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Enzymatic Deacetylation
Enzymatic deacetylation uses chitin deacetylases to perform a controlled, non-
degradable process that allows us to obtain a novel, well-defined chitosan. Such enzymes are
glycoproteins which are found in several insect species and fungi. They are extremely stable at
50 °C and show an excellent specificity for β- [1,4]-linked N-acetyl-D-glucosamine polymers.
Nevertheless, chitin substrates need to be pretreated so that the accessibility of the acetyl groups
to the enzyme can be improved [1].
The process and therefore the degree of deacetylation is determinant for the establishment
of chitosan characteristics. [16] Chitosan is a very interesting polymer as it is biodegradable,
non-toxic, biocompatible and has antibacterial effects, that can promote the acceleration of
wound healing. It has high adsorption, is capable of forming three- dimensional porous
structures, is easy to handle and has a low cost. Moreover, chitosan has mechanical stability,
especially when associated with collagen. [1, 4, 6, 10-12, 14, 18] It is a cationic polymer at low
pH (an exception since other polysaccharides are generally neutral or anionic) which gives it a
great affinity for anionic molecules. Therefore, chitosan can bond with mucous tissue
(mucoadhesiveness), being a good option for tissue engineering [1, 6, 10, 13-16]. In addition, its
hydrophilic surface promotes cell adhesion, proliferation, and differentiation and causes a
minimal foreign- body reaction on implantation. [7, 11, 17]
As mentioned above, the DA and MW influence, to a great extent, chitosan
characteristics like crystallinity, solubility, and its mechanical performance. This polymer has
low molecularweight and a high degree of deacetylation. As a result, chitosan shows a
reasonable solubility in water (that can be improved by chemical modification) and inorganic
acids. This last characteristic represents the major limit to chitosan preparation and uses for
biomedical applications. [1, 4, 6, 10, 14-16,18].
Due to these properties, chitosan has even more applications. It is very useful for drug
delivery, cancer diagnosis, immune regulation, treatment of damaged skin, nerve regeneration,
cartilage and bone repair, antioxidant properties and anti-inflammation. [3, 4, 11, 13, 15, 17]
Also, shows an antitumor and antidiabetic capacity and hemostatic effects (as its cationic nature
attracts the anionic blood cells and platelets allowing it to be used in postpartum hemorrhage,
dental surgery, and other surgical applications). [5, 19,20] Besides that, chitosan can interact
with substances like collagen, heparin, hyaluronic acid, gelatine, alginate and minerals: [21, 25]
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• Hydroxyapatite maximizes the in vivo osteogenic capacity which leads to an internal bone
growth with accelerated resorption of the matrix. (16, 21)
• Calcium aluminate also has an important role in bone regeneration (it releases Ca2+ that
increases osteogenesis and cementogenesis). In addition, it improves chitosan’s mechanical
strength, which is required for scaffolds. [21]
Despite all the information known about this biomolecule, the lack of clinical studies in humans
requires more and deeper studies on this matter.
Chitosan and its different morphologies Depending on environmental pH values and degree of
acetylation, chitosan can display different ways of production. At low pH (6.5), chitosan amines
are deprotonated and reactive, promoting interpolymer associations. These associations lead to
film, fibre, porous templates or hydrogel formation. [2, 20, 27, 28] It can also be presented as
sponge, powder, bead, solution, foams, microcapsules, tubes, membranous films, nanofibers,
scaffolds, nanoparticles. [2, 11, 16, 18, 22, 25].
Scientific literature supports that bacterias are unable to form biofilm by chitosan nanoparticles.
[24] Additionally, scaffolds are very useful in bone, skin (also regenerated using fibers and
hydrogels), liver, cartilage, nerve, and blood vessel wounds. Fibers and hydrogels allow
migration of inflammatory cells to the wound site and collagen matrix deposition. Furthermore,
hydrogels have the capacity to promote angiogenesis. [3]
Chitosan’s applications
4.1 Tissue Regeneration
Natural occurring biomaterials (like chitosan and collagen) and Calcium phosphate
ceramics (like hydroxyapatite) are commonly used for tissue regeneration. Chitosan, due to its
properties, is a great material to form different platforms where cells can adhere, grow in and
differentiate. [9, 10, 18]
Nevertheless, several studies proved that better results were achieved when chitosan was
combined with collagen in comparison with pure chitosan. [9, 16]
In addition, hydrogels can be used to produce scaffolds that have as advantages: minimal
physical flexibility, invasive procedure, and easiness of incorporating cells. However, one of its
disadvantages is that some cells have a lack of adhesion to scaffolds. Dhanya et al. prepared a
new scaffold, O-carboxymethyl chitosan (OCMC), as an attempt to solve this limitation. It has
high viscosity, good water solubility, and high hydrodynamic volume. [18]
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Moreover, chitosan-based platforms can also be used in a more delicate area, such as cardiac
patch for defects in myocardial tissue. Pok et al. prepared a gelatin-chitosan hydrogel-based
multilayer cardiac patch. This compound (which the main component is chitosan) increases
patch tensile strength and, due to gelatin, consolidates cell attachments. [18]
Another study has been developed to investigate in-vitro and in-vivo degradation rates of
porous freeze gelated chitosan (CH) and chitosan hydroxyapatite scaffolds. It is believed that
scaffold’s pore architecture is essential to an efficient cell seeding into the scaffold.
Simultaneously, it may offer enough space for the regeneration of newly formed tissue.
Therefore, those two characteristics should be taken in consideration when chitosan scaffold is
being produced. Biomaterial degradation rate may, also, be taken into account. It might be
sustained enough so that healthy tissue regeneration can be fulfilled [28].
Chitosan hydroxyapatite scaffolds are stable and biodegradable (however its degradation
rate is lower). Additionally, they provide an osteoconductive environment for bone formation
and have a high elastic modulus. These are the reasons why they are appropriate to bone
substitution and regeneration in dental implants [28].
On the other hand, scaffolds are favorable for tissue regeneration if they show a highly
porous structure to support cellular adhesion and proliferation and extracellular matrix (ECM)
production. Their degradation is dependent on pore morphology and size, porosity percentage,
surface area, and hydrophilicityPorous freeze gelated chitosan scaffolds have been tested in order
to be used for periodontal regeneration [28].
Peter et al. and Mota et al. concluded that degradation rate of chitosan can be reduced by
adding compounds (such as Bioglass™), what can contradict one of the major limitations of its
use for tissue regeneration. Therefore, the conclusions of these studies prove the enormous
chitosan and hydroxyapatite potential for use in composites that aim bone tissue engineering
applications [28].
Even so, the lack of precise information about chitosan’s characteristics, its large chemical
variability and studies’ methodological limitations make it difficult to reproduce and standardize
its clinical application. [10]
Wound healing
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Inflammation is a crucial part of healing, however, it encumbers tissue regeneration when

we are dealing with chronic wounds or biomaterial implantation. [3]
Until known, in order to decrease the inflammatory response, biomaterials have been
incorporated with anti-inflammatory therapeutics. Therefore, new scaffolds’ designs aim to
provide instructive signals for cell attachment, function and proliferation within the native tissue
and to modulate the immune response. [3]
Chitin-derived materials, such as chitosan, were discovered to be effective to chronic
inflammatory diseases treatment (inflammatory bowel disease, arthritis, sepsis, and asthma, for
example) as they are useful for drug delivery and in modulating immune responses (it attracts
macrophages and neutrophils to the wound, monitoring inflammatory response and
reepithelization). Their mechanism, although not fully defined, consists of molecules degradation
into monosaccharides, accelerating wound healing. [3, 18].
Nerve regeneration
Autografts have been used to treat peripheral nerve damage, but they have many
disadvantages. Therefore, chitosan is an excellent option to treat this condition as hydrophilicity
is considered to be essential for preventing fibrous scar tissue invasion and for promoting nerve
regeneration. Nerve conduits formed by this biopolymer have better mechanical properties and
slow biodegradability. [29]
Recently, porous hybrid membranes (such as chitosan- -glycidoxypropyltrimethoxysilane
- chitosan-GPTMS) were investigated for peripheral nerve regeneration. They were proven to
significantly improve nerve fibre regeneration and its functional recovery in rat models by
demyelination. The quantity of regenerated myelin fibres increased, as well as, myelin thickness
[29].
Neuronal disorders
Chitosan can play an important role in the prevention and treatment of age-related
diseases. Among others, it can prevent protein conformational diseases and treat those which
result from oxidative stress. Researchers from Fukuyama University conducted in vitro and in
vivo studies and concluded that, by decreasing oxidative stress (it suppresses the formation of
reactive oxygen species), chitosan can have a direct antioxidant effect in the systemic circulation.
[5, 19]
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Chitosan oligosaccharides (COS) are also used in order to treat neuronal disorders,
including Alzheimer’s disease, Parkinson’s disease, and nerve crush injury. Owing to their
shorter chain lengths, COS are rapidly soluble in water which confers them new properties like
being neuroprotective (such as β-amyloid and acetylcholinesterase inhibitory activities), anti-
neuroinflammatory and having anti-apoptosis effects. Moreover, Gong et al. explored the effects
of this polysaccharide on nerve regeneration after peripheral nerve injuries. As a result, they
concluded that it could enhance the quantity of regenerated myelinated nerve fibres, the cross-
sectional area of muscle fibres, the muscle action potentials and the thickness of regenerated
myelin sheaths in the nerves. [5]
Nowadays, neuroprotection aims to limit neuronal dysfunction or death after nerve
injury. For that, neuroprotective agents like antioxidants and anti-inflammatory agents are used.
The problem is that synthetic agents are linked to several side effects. Therefore, anti-neuronal
disorders agents with low toxicity have been studied. [5]
To conclude, Chitosan and its derivatives are neuroprotective through mechanisms like
antioxidative stress action; suppressing effect on Abeta aggregation; anti-neuroinflammatory
action; anti-apoptosis action; anti-excitotoxic action. [5]
CANCER
Cancer diagnosis
A cancer diagnosis is a crucial step since it gives some information about treatment
effectiveness. Theragnosis is a new conceptwhich combines molecular imaging (through
nanoparticles that are accumulated in tumor tissue) and anti-cancer drugs that are encapsulated
into nanoparticles. In this line of thought, chitosan is ideal for nanoparticle-baseddtheragnosis, as
it can be easily transformed into nanoparticles and self- degradable after accurate diagnosis and
drug delivery [6].
Cancer Therapy
In this field, chitosan has several applications, especially when prepared into
nanoparticles. This biomaterial can be an ideal vaccine adjuvant due to its cationic nature,
biocompatibility, safety and its ability to be used as an antigen carrier. It also showed a superior
immune activity when compared to traditional immunoadjuvants, as chitosan retains the peptide
antigen in the administration site for a longer period, allowing the antigen to be presented for
efficient immune activity [6]
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Chemotherapy
Anti-cancer drugs used in chemotherapy have several severe side effects, due to the low
specificity to the tumor tissue and their systemic administration. To minimize this non-
specificity therapy, selective embolization has been studied. It provides angiogenesis that will
affect cancer cells viability (it occludes arteries formed by those cells, inducing tumor’s cells
starvation). Chitosan is not a viable candidate to this approach since it is weak when produced
into microspheres. [18] Chemoembolization is a method that occurred later described by Park et
al. It combines the effect of embolization with sustained release of the anti-cancer drug. They
accomplished an efficient in vivo anti-cancer effect with a reduction of one-third of the initial
tumor size. [18]
Radiotherapy
Chitosan is soluble in acidic conditions, unlike neutral or basic ones. Yuka et al.
suggested that it can be chelated with several substances and then injected (as an acidic solution-
166Ho- chitosan complex) into human organs. It forms a solid structure that consists of a
radioactive pharmaceutical device, allowing radioactivity to be localized in the administration
site (when intratumorally and intrahepatically injected) [18].
Drug delivery
The presence of amino groups with positive charge allows chitosan to promote chemical
and physical cross-linking (chitosan can adhere to negatively charged organs’ surface), which is
the main advantage of its use. In addition, if chitosan is modified with hydrophobic groups, it can
be used to encapsulate hydro-phobic drugs, since it forms intra- and intermolecular interactions
[1, 18].
Therefore, chitosan can be a successful carrier for many different therapeutic agents:
proteins, peptides, growth factors, DNA, vaccines, anti-inflammatory, antibiotics, and other
drugs for parental and non-parental routes of administration (like oral, ocular, topical, nasal,
transdermal and pulmonary) [27]. Although the many forms of chitosan formulations, the choice
to use a micro or a nanoparticle depends on what is required. For example, since microparticles
cause obstruction in the blood vessels, nanoparticles are the best choice for intravenous delivery.
On the other hand, if nanoparticles are used to pulmonary delivery (to treat different pulmonary
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infections and diseases, like tuberculosis) they would be exhaled. So, in this case, the use of
microparticles would be better [1, 27].
Both chitosan’s micro and nanoparticles, when loaded with antibiotics and administrated
by the oral route, can be used in the treatment of gastrointestinal and systemic diseases, such as
ulcerative colitis, irritable bowel syndrome, Crohn’s disease and suppression of Helicobacter
pylori. Drug colon targeting is useful since the degradation of chitosan is done by the specific
local microflora [27].
Bone regeneration
Bone regeneration is important to orthopedic, maxillofacial, and oral surgery (such as in
periodontology and implantology field) in order to correct the bone defects. Therefore, the
density and bone volume must be recreated, without forgetting the function. At the moment,
therapies use bone tissue’s grafts from different kind of origins – heterologous, homologous or
autologous (each one of them with advantages and disadvantages). As autologous grafts (which
have no risk of rejection and diseases transmission) are limited, a new approach for bone
regeneration is needed. [7, 8, 12, 13, 16, 21]
Furthermore, the biomaterials have been studied because of their capacity to induce a
biological response, knowing that the results obtained are satisfactory. Nowadays, guided bone
regeneration (GBR) is a therapy for alveolar bone repair using a membrane which has the ability
to prevent the formation of fibrous connective tissue [7, 8, 21, 22]. The membrane has some
properties like bioactivity (promotes cellular migration, proliferation, and infiltration),
biocompatibility, bioresorption, lack of cytotoxicity and stability. This membrane can be
combined with natural polymers (collagen and chitosan). The degradation rate of polymers is
also a crucial factor: it should be total and not require a second surgical intervention. That is why
it may have a long-term influence on the success of bone regeneration [7, 12, 22].
According to Jiayu Zhang et al. [22] local drugs delivery systems (DDS) allow aspirin to
be in the right place at the correct moment. As a result, there will be a reduction in the number of
administrations, bacterial resistance, gastrointestinal effects and we can avoid variations of large
concentration. [22]
Non-steroidal anti-inflammatory drugs (NSAID) are reported to be effective in: repair of
bone defects and fracture (bear in mind that dose is an important factor), reduce the percentage
of tissue destruction and bone loss in periodontal disease [22].
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The results show a good controlled-release pattern, the biocompatibility and osteogenic
potential of the membranes and also a significant increase of bone, in animal models. [22] On the
other hand, GBR membranes are vulnerable to infection because of their exposure to pathogenic
agents, which affects bone regeneration. In an attempt to counteract this, it is used antibiotic
ointments. However, it should be created a new antibacterial GBR membrane. (8) Shiqing Ma et
al. [8] used membranes containing antibacterial agents such as tetracycline, chlorhexidine,
minocycline, and amoxicillin. These ones have the ability to inhibit bacterial proliferation and
induce the growth of fibroblasts and osteoblasts, as well as, stimulate angiogenesis. [8]
An asymmetric collagen/chitosan GBR membrane (CCM membrane) was carried with
encapsulated minocycline. This membrane had loose chitosan (that allows it to be
osteoconductive) and dense collagen layers (act as a physical barrier), low degradation and
activity against Fusobacteriumnucleatum and Porphyromonasgingivalis, which are the core of
periodontitis disease [8].
CCM membrane promoted new bone formation. In addition, there weren’t verified
inflammatory reactions and grade of degradation has raised. Furthermore, it was observed a huge
number of mesenchymal stem cells and capillary vessels (responsible for transportation of
nutrient and stem cells), which is an ideal environment of osteoinduction [8] Jae Min Song et al.
[7]. Reinforced that chitosan/fibrin–hydroxyapatite (CFB–HAP), a membrane that has also been
studied, promotes haemostasis and adhesiveness and is biocompatible, anti-infective, resorbable
and plastic. [7] After this study, it is possible to affirm that the bone repair capacity is similar
between CFB–HAP membrane and collagen membrane. In addition, the absorbable chitosan
membrane can be used as a barrier membrane in GBR. However, the inflammatory reaction
didn’t existin the CFB-HAP, which can comprise the success of the guided bone regeneration
[7].
Antimicrobial activity
The endodontic treatment has a significant percentage of failure, associated with
inefficient decontamination of root canals and repair of changes caused by the disease [24].
In this context, antimicrobial photodynamic therapy (PDT) has been used as it promotes cross-
linking between collagen and proteins and has a large spectrum of antimicrobial activity. Also,
some nanoparticles like chitosan, which is able to reduce the bacterial-biofilm, have been
modified with a photosensitizer (such as rose Bengal, that produces synglet oxygen) in order to
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highlight PDT. As a result, bioactive polymeric chitosan nanoparticles functionalized with rose-
bengal (CSRBnp) were studied to eliminate biofilms and stabilize the matrix (dentin-collagen)
attending to its positive charge and nano size. Those characteristics, also promote a Physico-
chemical interaction with some other particles [24].
The conjugation of this cationic molecule with photosensitizer allowed the penetration on
the bacterial membrane. These cells were efficiently killed, as well as their biofilm structure
destroyed. The antibacterial effects of chitosan are based on promoting bacterial membrane
damage, increasing its permeability and providing intracellular leakage [24]. In addition,
chitosan nanoparticles have been incorporated into root canal sealers in order to reduce the
biofilm formation and inhibit microbial penetration in the dentin-root [13].
In another view, pulp exposure can be treated without chemical injury to pulp cells. For
that, it was tested a chitosan/collagen membrane combined with calcium-aluminate
microparticles. The results were: an increase of human dental pulp cells proliferation, trailed by
odontoblastic phenotypes (high expression) and intensive deposition of matrix’s mineralized
[30].
Implants
The tooth extraction is associated with resorption of alveolar bone in a period of three
months. Residual ridge absorption is a real problem to posterior rehabilitation of the edentulous
area because of the lack of bone support. Any solution present has to promote osteogenesis and
also angiogenesis. [23]
The core is to preserve the level of bone in order to realize a functional and aesthetic
restoration. A dental implant is used to replace multiple or single missing teeth or to support a
prosthesis. The lifetime of a dental implant is limited by its capacity to uptake and hold
backwater, which is related to diffusion’s coefficient of the material. [23, 28].
One of the proposals is to use Poly (L -lactic-co-glycolic) acid and nanohydroxyapatite together
with chitosan to delivery adrenomedullin (ADM). It was reported that ADM is a substance which
promotes the mitogen activity and proliferation of osteoblasts. Besides that, it seems like a
promising new formula for bone regeneration, once it increases the formation of new bone tissue
and is capable of stabilizing the residual alveolar ridge [13, 23].
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Oral diseases
It is known that chitosan has activity against oral pathogens such as Prevotellaintermedia,
Porphyronomasgingivalis, and Actinobacillusactinomycetemcomitans, inhibiting the bacterial
plaque formation. To some authors, this inhibition can occur as chitosan nanoparticles kill the
bacteria or have a direct action in the bacteria-substrate. Additionally, it has been reported that
the collagen’s turnover has increased when chitosan is present. In this line of thought, toothpaste
could contain chitosan nanoparticles, in order to reduce bacterial biofilm formation [13, 17].
Carie (Decay)
Chitosan nanoparticles can be used to remineralize dentine and enamel caries. This
occurs because chitosan’s nanoparticles, at a neutral pH, cause damage in cariogenic
microorganisms, such as S.mutans’ cells. [13] There are studies using nanocomplexes of
carboxymethyl chitosan/amorphous calcium phosphate (CMC/ ACP) to remineralize the dentine
via biomimetic strategy. This remineralization is more difficult to occur due to dentine’s organic
matrix when it is compared to enamel. In addition, the remineralization of dentine happens by its
residual crystals’ growth. Using a single-layer model, CMC/APC released ACP nanoparticles to
achieve intrafibrillar mineralization forms of collagen (the minerals are deposited in the collagen
fibril’s gap zones and microfibrillar space) - bottom-up strategy. The remineralization of
demineralized dentine happens preserving the greatest extent possible of dentinal tissue when
this combination is used [31].
Candida
In the last few decades, the number of cases with microbial infections increased
exponentially, possibly due to the raise of immunosuppressive diseases or infections (eg.
leukemia or AIDS), as well as secondary effects from cancer chemotherapy. Moreover, the
resistance of pathogenic microorganisms to conventionaltreatment has also increased due to
primary or secondary resistance [14]. Chitosan acts against the growth of hyphal, filamentous
fungi and yeast infection (human), as well as spore germination. The plasma membrane of fungi
(phospholipid heads and glycoinositolphosphorylceramides), that is composed of
polyunsaturated fatty acids, is the target of chitosan. [14] It is important to take into
consideration that the more fluid the membrane is, the more sensitive will be to the fungicides.
Therefore, some fungi are resistant to chitosan because they have a low concentration of
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polyunsaturated fatty acids, such as P. chlamydospores, nematophagous and entomopathogenic

fungi [14].
Chitosan can be used against Candida spp. and has the potential to, in the future, reduce
the use of antifungals, such as amphotericin B and fluconazole [14]. Chitosan-coated
nanoparticles (CSNPs) containing fluconazole (FLZ) were tested for local treatment. The results
show that the CS- NPs interact with mucins, extend the delivery of the drug and promote
animals’ recovery from candidiasis. FLZ remains in the mucosa to act locally and it isn´t
absorbed to the systemic circulation. [18, 32] Due to this, this formulation hasn´t a cytotoxic
effect and can eliminate the chance of drug interaction when is compared to systemic therapy
with FLZ [32].
Periodontal disease
Although numerous treatments are used to regenerate the alveolar bone, cementum, and
loss of periodontal ligaments (PDL), new approaches are required to treat periodontal lesions
[17].
Porous membranes of chitosan, combined or not with hydroxyapatite, were prepared
using freeze gelation (FG). They were to be used for GTR (guided tissue regeneration) to
increase periodontal recovery. They are unique due to their physicochemical properties,
ultrastructural morphology, degradation, mechanical properties and biocompatibility [20].
In this study, it was demonstrated that porous membranes would be used as a core layer
and that the selection of solvent and quantity of incorporated HA benefited the properties
(chemical and physical) of membranes. Additionally, it was observed the resilience of the
membranes while they were handling in wet and dry conditions. The cellular response was
positive, which indicates that cellular activity was affected by HA incorporation. [20] Some
other applications of chitosan are succinctly explained in the table below
CONCLUSION
In this review, the applications of the biopolymer chitosan were highlighted in the
medical field, including dentistry. Thanks to its properties, such as biocompatibility and
biodegradability, its cationic nature, it's non- toxic and antibacterial effects and the fact that
causes a minimal foreign body reaction, chitosan is a useful biomaterial in drug delivery,
immune regulation, tissue regeneration, cancer diagnosis, among many other. Particularly in
dentistry, it has application in bone regeneration, alveolar bone repair and in the implantology
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field. It’s also very promising in endodontic treatment (since it has an antibacterial action), decay
remineralization and treatment of fungal diseases and periodontal lesions.
In the near future, this innovative biomaterial, which research has been increasing
exponentially in the last years, can be very helpful in the diagnosis and treatment of many
prevalent diseases. Nevertheless, more studies are needed, in order to its mechanism of action
and applications are fully understood and known
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ANTIMICROBIAL ACTIVITY OF PLANT-MEDIATED SYNTHESIS OF SILVER

NANOPARTICLES
S.D.K.Shri Devi, Sudha and Monika

Department of Botany , Sri Sarada College for women (Autonomous), Salem, Tamil Nadu
Email: saishri28@gmail.com
Department of Botany, Chaudhary Bansi Lal University, Bhiwani (Haryana),
Email: sudhayadav1111@cblu.edu.in
Department of Botany, Chaudhary Bansi Lal University, Bhiwani (Haryana)
Email: moni.ydv104@cblu.edu.in
ABSTRACT
Interest in “green nanotechnology” in nanoparticle biosynthesis is growing among
researchers. Nanotechnologies, due to their physicochemical and biological properties, have
applications in diverse fields, including drug delivery, sensors, optoelectronics, and magnetic
devices. This review focuses on the green synthesis of silver nanoparticles (AgNPs) using plant
sources. Green synthesis of nanoparticles is an eco-friendly approach, which should be further
explored for the potential of different plants to synthesize nanoparticles. The sizes of AgNPs are
in the range of 1 to 100 nm. Characterization of synthesized nanoparticles is accomplished
through UV spectroscopy, X-ray diffraction, Fourier transform infrared spectroscopy,
transmission electron microscopy, and scanning electron microscopy. AgNPs have great
potential to act as antimicrobial agents. The green synthesis of AgNPs can be efficiently applied
for future engineering and medical concerns. Different types of cancers can be treated and/or
controlled by phytonanotechnology. The present review provides a comprehensive survey of
plant-mediated synthesis of AgNPs with specific focus on their applications, e.g., antimicrobial,
antioxidant, and anticancer activities.
KEYWORDS: Green synthesis, Silver nanoparticles, Optimization, Characterization,
Biomedical applications.
INTRODUCTION
The utilization of nanotechnology for constructing nanoscale products in research and
development divisions is growing [1]. Nanotechnology can be used to produce a broad range of
products applicable to an equally broad array of scientific sectors. “Creation,” “exploitation,”
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and“synthesis” are terms associated with nanotechnology,which generally considers materials

that measure less than 1 mm. “Nano” is derived from the Greek word “nanos”, meaning “dwarf,
tiny, or very small” [2]. Nanotechnologies are generally classified as wet, dry, and
computational. Wet nanotechnology is associated with living organisms such as enzymes,
tissues, membranes, and other cellular components. Dry nanotechnology is associated with
physical chemistry and the production of inorganic items, such as silicon and carbon.
Computational nanotechnology is associated with simulations of nanometer-sized structures[3].
These three dimensions (wet, dry, and computational) depend on each other for optimal
functionality, represented in Fig. 1. Nanotechnology supports diverse unique industries, such as
electronics, pesticides, medicine, and parasitology, and thus provides a platform for collaboration
[4]. Nanobiotechnology provides one such example, wherein the study and development
combine multiple scientific sectors, including nanotechnology, biotechnology, material science,
physics, and chemistry [2, 5].
Biologically synthesized nanoparticles with antimicrobial, antioxidant, and anticancer
properties are possible through the collaboration of different natural science sectors. These
nanotechnologies may provide novel resources for the evaluation and development of newer,
safer, and effective drug formulations [6].
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Infections caused by microorganisms during the treatment of orthopedic diseases,

resulting in faster bone recovery [7]. At present, varied physical, chemical, biological, and hybrid
methods (Fig. 2) are utilized to synthesize distinct nanoparticles [8, 9]. The synthesis of
nanoparticles has traditionally relied on two approaches, physical and chemical. These
approaches include ion sputtering, solvothermal synthesis, reduction, and sol-gel techniques.
Nanoparticle synthesis methods can also be classified as bottom-up and top-down. Chemical
methods involve the reduction of chemicals [10], electrochemical procedures [11], and reduction
of photochemicals [12]. Plant-based synthesis of nanoparticles is in contrast faster, safer and
lighter; works at low temperatures; and requires only modest and environmentally safe
components [13]. Plant-based nanoparticles have attracted more attention due to growing interest
in environmentally conscious products. In addition, the synthesis of nanoparticles using plants
offers other advantages [14]. Furthermore, physical requirements for their synthesis, including
pressure, energy, temperature, and constituent materials, are trivial.
Nanoparticles made of noble metals have also received attention over the last few years,
as they can be used in medicine, biology, material science, physics, and chemistry [15]. Among
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the several noble metal nanoparticles, silver nanoparticles (AgNPs) have attracted special
attention due to their distinct properties, which include favorable electrical conductivity,
chemical stability, and catalytic and antibacterial activity [12]. Silver at the nanoscale also has
different properties from bulk silver. Synthesis of AgNPs is an emerging area and is much sought
after [16]. The green synthesis of AgNPs has been accomplished using plants, microorganisms,
and other biopolymers [12]. Wet chemical synthesis can be robustly scaled for the large-scale
synthesis of AgNPs of tunable shape and size through optimization of synthesis conditions.
However, wet chemical methods use toxic chemicals, which are hazardous for the
environment and usually result in the adsorption of toxic chemicals on to the surface of
synthesized AgNPs, making them unsuitable for biomedical applications. In contrast, physical
methods are expensive and cumbersome for the largescale production of nanoparticles.
Therefore, the development of environmentally conscious, energy-efficient, facile, and rapid
green synthesis methods that avoid toxic and hazardous chemicals has attracted significant
interest [17]. In addition, due to their potent antimicrobial activity, AgNPs have also been used in
clothing [18], foods [19], sunscreens, and cosmetics [20, 21]
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SYNTHESIS OF AGNPS
The use of plants for nanoparticle synthesis offers a wide range of benefits over other
biological synthesis methods because it does not require the maintenance of cell cultures and
incorporates support for the large-scale synthesis of nanoparticles [22]. Extracellular
nanoparticle synthesis, which utilizes extracts from individual leafs rather than entire plants, may
prove to be more inexpensive due to easier downstream processing (Fig. 3). Sastry and his group
are responsible for pioneering nanoparticle synthesis using plant extracts [22–27].
Green synthesis of AgNPs using plant extracts containing phytochemical agenhas
attracted considerable interest (Table 1). This environmentally friendly approach is more
biocompatible and cost-efficient and includes the capability of supporting larger synthesis [28,
29]. The synthesis of AgNPs via different “green” chemico-physical conditions, as well as by
numerous microorganisms, has been heavily investigated. When AgNPs are chemically
synthesized, three main components are required: (1) silver salt (e.g., AgNO3), (2) a reducing
agent (e.g., NaBH4), and (3) a stabilizing or capping agent (e.g., polyvinyl alcohol) for
controlling the size of nanoparticles and preventing their aggregation [30]. AgNPs have
applications in wound-healing, eye disease therapy, DNA processing, and pharmaceuticals in
additionto other relevant mainstream applications: electronics, optics, catalysis, and Raman
scattering [31–35]. Lokini et al. [36] showed that AgNPs could destabilize the outer membrane
and rupture the plasma membrane, thereby depleting intracellular ATP. Silver has a greater
affinity to react with sulfur or phosphorus-containing biomolecules in the cell; therefore, sulfur-
containing proteins in the membrane or inside cells and phosphoruscontaining elements like
DNA are likely to be preferential sites for binding AgNPs. The advantages of using plants for the
synthesis of nanoparticles include their availability, safety in handling, and presence of a
variability of metabolites that may aid in reducing silver. The time required to reduce 90 % of
silver ions is approximately 2 to 4 h [27]. Gericke and Pinches [37] reported that the size of
particles that form intracellularly could be controlled by altering key factors such as pH,
temperature, substrate concentration, and time of exposure to the substrate.
The biochemical and molecular mechanisms of AgNP biosynthesis remain poorly
characterized and should be investigated to further optimize the process. For instance,
characterization of biochemical mechanisms underscored the importance of phytochemicals,
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which may mediate biosynthesis. Improvements in chemical composition, size, shape, and
dispersity of nanoparticles would permit the use of nanobiotechnology in a variety of other
applications [38]. Plant crude extracts contain novel secondary metabolites such as phenolic acid,
flavonoids, alkaloids, and terpenoids, which are mainly responsible for the reduction of ionic
metal into bulk metallic nanoparticles [39]. Primary and secondary metabolites are constantly
involved in redox reactions required to synthesize ecofriendly nanoparticles,
Table 1 Green synthesis of silver nanoparticles using different plant extracts (Continued)
Azadirachta indica Leaf 20 Triangular [159]
Ocimum tenuiflorum Leaf 50 Cuboidal [159]
Artocarpus heterophyllus Seed 10.78 Spherical and irregular [71]
Cocos nucifera Coir 22 Spherical [160]
Eucalyptus chapmaniana Leaf 60 Spherical [52]
Morinda citrifolia Root 30–55 Spherical [161]
Thuja occidentalis Whole plant 122 Spherical [162]
Hydrastis Canadensis Whole plant 111 Spherical [162]
Phytolacca decandra Whole plant 90.87 Spherical [162]
Biosynthesis reactions can be modulated to transform the shape and size of nanoparticles
by using different metal concentrations and amounts of plant extract in the reaction medium [27,
40]. Capsicum annuum leaf extracts contain a number of biomolecules, such as proteins,
enzymes, polysaccharides, amino acids, and vitamins, which could act as bioreductants for metal
ions or as scaffolds to direct the formation of AgNPs in solution. In detail, the mechanism
underlying the bioreduction of silver was hypothesized to first involve trapping of silver ions on
the surface of proteins in the extract via electrostatic interactions (i.e., recognition process).
Silver ions are then reduced by proteins, lead in changes in their secondary structure and the
formation of silver nuclei. Silver nuclei subsequently grow by the further reduction of silver ions
and their accumulation at nuclei [41]. Callicarpa maingayi stem methanolic extracts were used
for the synthesis of AgNPs, leading to theformation of [Ag (Callicarpa maingayi)]+ complex
Plant extracts contain aldehyde groups, which are responsible for the reduction of silver ions into
metallic AgNPs. The different functional group, –C = 0, C = N, indicates amide I of polypeptides
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that are responsible for the capping of ionic substances into metallic nanoparticles. Molecular
studies on the biosynthesis of silver crystals have revealed a complex process, which is not fully
understood yet [42].
PHYSICAL REQUISITES FOR THE SYNTHESIS OF AGNPS
Easier, more reliable, and environmentally friendly methods to synthesize nanoparticles
accelerate their widespread adoption, which would benefit humans and the environment [43].
Silver disassembles into particles following the addition of plant extract, which may lead to a
color change. Solutions of AgNPs appear dark, yellow-brown in color because of the surface
plasmon resonance phenomenon [44]. Gardea-Torresdey et al. [45] determined the influence of
pH on the mass of nanoparticles when using alfalfa biomass in the biosynthesis of colloidal gold.
Mock et al. [46] reached a similar conclusion that the unique pH conditions of different extracts
affect nanoparticle size and shape. Two extracts from the same host plant may have a different
pH, thus highlighting the need for better synthesis methods for nanoparticles. Large
nanoparticles are most often formed only at lower pH values, instead of higher pH values, as has
been previously reported [47, 48]. Dwivedi and Gopal [49], utilizing extracts of Chenopodium
album, observed trivial variations in zeta potentials of nanoparticles in pH conditions ranging
from 2 to 10 and determined that nanoparticles were more stable when exposed to higher pH
conditions.
Veerasamy et al. [50] demonstrated that mangosteen extracts induced the nucleation of a
cluster of AgNPs at pH values over 4. Furthermore, nanoparticles grew rapidly, with their pH
values ranging from basic to neutral. These results demonstrate the significant impact of pH on
parameters of nanoparticles. The formation and growth of nanoparticles is retarded by acidic
conditions, whereas basic conditions promote nanoparticle assembly. Larger nanoparticles are
formed in lower pH conditions (pH 4), whereas significantly smaller nanoparticles are formed in
higher pH conditions (pH 8). Our results indicate that the size of nanoparticles decreases when
pH increases. pH values in the range of 2–14 play an important role in the synthesis of AgNPs.
In plants, AgNP synthesis occurs at various pH values depending on the plant species [50].
However, previous studies have indicated that neutral pH is optimal for AgNP synthesis.
At this pH, little or no assembly of AgNPs into particles of suitable size and shape occurs [51].
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Newly synthesized AgNPs, formed within 60 min of incubation with leaf extracts of Eucalyptus
chapmaniana, exhibited a UV-Vis peak at 413 nm [52]. The UVVis spectra of AgNPs,
synthesized with leaf extracts of Desmodium gangeticum for an optimum incubation time of 90
min, exhibited a peak at 450 nm [53]. VilchisNestor et al. [54] demonstrated a UV-Vis peak at
436 nm for AgNPs formed within 4 h of incubation with Camellia sinensis extracts. Chandran et
al. [27] synthesized AgNPs with leaf extracts of Aloe vera incubated for 24 h, which exhibited a
UV-Vis peak at 410 nm. UVVis absorption spectra reach a maximum when the synthesis
of nanoparticles (NPs) is complete, which requires sufficient time for the nucleation and
subsequent stabilization ofnanoparticles Song and colleagues synthesized stable AgNPs
extracellularly, with average particle sizes ranging from 15 to 500 nm, with Pinus densiflora,
Diospyros kaki, Ginkgo biloba, Magnolia kobus, and Platanus orientalis leaf extracts. The rate of
synthesis and final conversion to AgNPs was faster with higher reaction temperatures. However,
average particle sizes of nanoparticles produced with D. kaki leaf extracts decreased from 50 to
16 nm when the temperature of synthesis was increased from 25 to 95 °C [16]. Ocimum sanctum
leaf extracts could reduce silver ions into crystalline AgNPs (4–30 nm) within 8 min of the
reaction. These nanoparticles were likely stable due to the presence of proteins, which may act as
capping agents. O. sanctum leaves contain ascorbic acid which was likely important for the
reduction of silver ions into metallic AgNPs [55].
Monodisperse spherical AgNPs (~3 nm) were also synthesized using gum kondagogu
(nontoxic polysaccharide derived as an exudate from the bark of Cochlospermum
gossypium)[56]
CHARACTERIZATION OF SYNTHESIZED AGNPS

The synthesis of AgNPs using a 5:1 ratio of fruit extracts of Piper longum was evident by
a change in the color of 1 mM AgNO3 solution from colorless to brownishyellow, which
resulted in a peak at 430 nm in UV-Vis spectra [57]. Aqueous leaf extracts of Manilkara zapota
were used to synthesize AgNPs, which exhibited XRD with 2θ values of 38.06°, 44.37°, 64.51°,
and 77.31° sets of lattice planes, which may be indexed to the (111), (200), (220), and (311)
face-centered cubic (fcc) structure of silver, respectively [58].
Fourier transform infrared (FTIR) spectroscopy, which is used to evaluate chemical
bonds in surface atoms and functional atoms on the surface of nanoparticles, can be used to
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characterize physical properties of nanomaterials and their functions [59, 60]. Certain proteins
and metabolites, such as terpenoids or flavonoids that are present in leaf extracts of Prosopis
juliflora, may be responsible for the decay and pause of AgNPs synthesis [61]. The FTIR spectra
of AgNPs synthesized using either fresh or dried Codium capitatum extracts exhibited a strong
transmission band at 1535 cm−1 corresponding to the bending vibration of secondary amines of
proteins. The FTIR peak at 1637 cm−1 for AgNPs synthesized using Andrographis paniculata
extracts can be attributed to the carbonyl stretch of amides and could be related to proteins that
potentially cap AgNPs [62]. In C. annuum extracts, the formation of AgNPs is mediated by
amine groups or the secondary structure of proteins [63].
The hydroxyl and carbonyl groups present in carbohydrates, flavonoids, terpenoids, and
phenolic compounds are powerful reducing agents that may be responsible for the bioreduction
of Ag+ ions necessary for AgNP synthesis. FTIR studies confirm that the carbonyl groups of
amino acids and peptides of proteins have a strong affinity to bind metal ions, and they may
encapsulate nanoparticles, forming a protective coat-like shell that prevents their further
aggregation and leading to their stabilization in the medium [64].
A single-step method (biogenic) for the synthesis of AgNPs utilizes Ziziphus jujuba leaf
extract as a reducing and stabilizing agent at room temperature. TEM images revealed
nanoparticles featuring differing shapes and sizes, averaging 25 nm. These results were
confirmed by DLS analysis, which revealed a hydrodynamic radius of 28 nm [65].
Environmentally friendly synthesis of AgNPs, which utilized Argemone mexicana leaf extracts
and were 20 nm in size, had antimicrobial and antifungal activity against multiple bacterial and
fungal pathogens [66]. Extracts from the Cycas leaf were utilized to prepare AgNPs measuring
2–6 nm [67]. AgNPs, measuring 14 nm and synthesized using Solanum torvum extracts,
exhibited a peak at 434 nm in UV-Vis spectra. Using EDX analysis, Arunachalam et al. [28]
showed that AgNPs were crystalline in nature and observed strong signal energy peaks for silver
atoms in the range of 2–4 keV with weaker signals for carbon, oxygen, and chloride, which are
prevalent biomolecules in Memecylon umbellatum. The size, shape, and size distribution of
nanoparticles were observed by TEM and selected area electron diffraction (SAED) patterns of
TEM images [68]. The crystalline nature of AgNPs was determined by SAED, which revealed
fcc silver.
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A facile biosynthesis method utilizing methanolic extracts of Vitex negundo, which can
be performed at room temperature, was used to successfully synthesize spherical colloidal
AgNPs of different sizes, although it required different reaction times. The sizes of colloidal
AgNPs prepared for 6, 24, and 48 h averaged 10.11 ± 3.98, 12.80 ± 4.97, and 18.23 ± 8.85 nm,
respectively [69]. The morphology and size of AgNPs synthesized using Pulicaria glutinosa
extracts were examined by TEM, which revealed monodisperse spherical nanoparticles between
40 and 60 nm [70]. AgNPs synthesized using seed extracts of Artocarpus heterophyllus exhibited
variance in their size, ranging from 3 to 25 nm with an average of 10.78 nm [71]. AgNPs
fabricated using Boerhaavia diffusa leaf extracts as the nontoxic reducing agent and examined by
TEM revealed AgNPs that were fcc structures of spherical shape and an average particle size of
25 nm [72].
BIOMEDICAL APPLICATIONS OF AGNPSANTIMICROBIAL ACTIVITY OF AGNPS

The threat posed by the potential outbreak of antibioticresistant microbes is growing
globally and demands the introduction and production of novel more advanced platforms for the
study and development of more potent antimicrobial agents against multidrug-resistant strains
[73]. The antimicrobial activity of AgNPs is widely recognized, though their activity can change
with physical characteristics of the nanoparticle, such as its shape, mass, size, and composition,
and conditions of its synthesis, such as by pH, ions, and macromolecules [74].
Their shapes can be relevant to their antibacterial activity [75]. Compared to larger
AgNPs, smaller AgNPs have a greater binding surface and show more bactericidal activity [76].
Variation in the thickness and molecular composition of the membrane structures of
grampositive and gram-negative bacteria account for the difference in their sensitivities to
AgNPs [77]. Bactericidal activity is presumably due to changes in the structure of the bacterial
cell wall as a result of interactions with embedded AgNPs, leading to increased membrane
permeability and consequently death [78]. AgNPs also interact with sulfur- and phosphorus-rich
biomaterials, which include intracellular components, such as proteins or DNA, and extracellular
components such as membrane proteins. These components influence the respiration, division,
and ultimately survival of cells [79]. Upon compromising the bacterial cell wall, silver ions (as
part of AgNPs) can enter into cells, leading to the accumulation of damaged DNA and effect on
protein synthesis [80].
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AgNPs synthesized with Skimmia laureola leaf extracts have antibacterial activity, with
maximum growth inhibition activity against Staphylococcus aureus (14.67 mM), followed by
Klebsiella pneumoniae, Pseudomonas aeruginosa (14.33 mM), and Escherichia coli (11.67 mM)
[81]. AgNPs synthesized with mangrove plant Avicennia marina extracts exhibited highest
inhibition activity against E. coli (18.40 ± 0.97 mM) and lowest against S. aureus (10.87 ± 1.33
mM). Its minimum inhibitory concentration (MIC) and minimum bactericidal concentration
(MBC) were 0.25 and 50.0 μg/mL, respectively, against select bacteria [82]. Sankar et al. [83]
synthesized AgNPs with extracts of oregano (Origanum vulgare), which exhibited antimicrobial
activity against human pathogens, including Escherichia coli, Aeromonas hydrophila, Salmonella
spp., Shigella dysenteriae, Salmonella paratyphi, and Shigella sonnei. Furthermore, AgNPs were
cytotoxic to human lung cancer lines (A549 cells), which killed 50 % of cells at 100 μg/mL.
Sathishkumar et al. [84] evaluated the bactericidal activity of AgNPs synthesized with Morinda
citrifolia leaf extracts against a wide range of human pathogens, such as Escherichia coli,
Pseudomonas aeruginosa, Klebsiella pneumoniae, Enterobacter aerogenes (gram-negative),
Bacillus cereus, and Enterococcus sp. (gram-positive).
The antibacterial activity of AgNPs synthesized with Rosmarinus officinalis extracts was
tested against gram-positive bacteria, and the maximum zones of inhibition at dosages of 20, 40,
and 80 mg/disk were 21.52, 30, and 31.2 mm, respectively, against S.aureu
and 13.4, 15.63, and 16.21 mm, respectively, against Bacillus subtilis [85]. The antimicrobial
activity of the medicinal plant Onosma dichroantha and antimicrobial activity of silver chloride
nanoparticles suggest a novel approach to the development of bactericides applicable to a wide
range of applications, such as the treatment of burn wounds and injuries [86].
AgNPs synthesized with Prunus yedoensis leaf extracts exhibited significant antibacterial
activity against two skin pathogens, Propionibacterium acnes and Staphylococcus epidermidis.
Exhibited zone of inhibition (ZOI) sustained greater levels of AgNPs (30 μg) after 48 h of
inhibition against two gram-positive bacteria; furthermore, tetracycline sulfate at volume of 100
μg/mL was evaluated [87]. AgNPs synthesized with O. vulgare leaf extracts have broad-
spectrum antibacterial activity against nine different human pathogens. Greater than 10-mm
zones of inhibition were observed against Escherichia coli (enteropathogenic,EP), Aeromonas
hydrophila, Salmonellaparatyphi, Salmonella sp., Shigella dysenteriae, and Shigella sonnei. This
level of antibacterial activity was comparable to the standard antibiotic chloramphenicol [83].
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Saxena et al. [88] synthesized AgNPs using Ficus benghalensis leaf extracts, and its
bactericidal activity against E. coli was evaluated by the broth microdilution method. The
bactericidal activity of AgNPs synthesized with B. diffusa plant extracts was evaluated against
three fish bacterial pathogens, Aeromonas hydrophila, Pseudomonas fluorescens, and
Flavobacterium branchiophilum. Of these, F. branchiophilum was more sensitive to AgNPs, and
the other two pathogens were equally sensitive [72]. Tripathi et al. [89] evaluated bactericidal
activity of silver nanoballs at a concentration of 40 μg/mL against Escherichia coli, Salmonella
typhimurium, Bacillus subtilis, and Pseudomonas aeruginosa by measuring colony-forming units
(CFU). Silver nanoballs prevented the growth of bacteria and induced toxicity.
AgNPs, which are filled with polyphenolic compounds, disrupt the cell walls of bacteria,
which make gramnegative bacteria specifically sensitive. Polyphenolic compounds generate free
radicals and other oxygenbased reactive species, which can induce considerable damage and
toxicity [75]. Other damages may result as membranes become disrupted, Including the
widespread loss of K+ ions, leading to a decrease in membrane potentia
Significant membrane disruption results in cytoplasmic leakage, which includes the
discharge of proteins and lipopolysaccharide molecules. The outer membrane of bacteria is
composed of lipopolysaccharides and is fundamentally asymmetric, while the inner membrane
comprises tight chains of phospholipids, which are semi-permeable [75].
The exact mechanism of interaction between AgNPs and bacteria is not fully understood.
AgNPs may attach to the cell wall and thus disrupt membrane permeability and ultimately cell
respiration. AgNPs can also directly penetrate into cells since they may bind to cell wall proteins
that contain sulfur and phosphorus-containing biomolecules such as DNA [90, 91]. Thus, they
can easily bind to constituents of the bacterial cell and disturb normal functions of the cell.
Another possible mechanism is the release of Ag cations, which are antibacterial, from AgNPs
[92].
AGNPS IN CANCER CONTROL
AgNPs perform well as cancer therapeutics because they can disrupt the mitochondrial
respiratory chain, which induces the generation of reactive oxygen species (ROS), and ATP
synthesis, which can induce DNA damage [93, 94]. AgNPs synthesized with Sesbania
grandiflora leaf extracts were demonstrated to be cytotoxic to MCF-7 cancer cells.
Morphological characteristics, including the disruption of membrane integrity, decreased cell
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growth, cytoplasmic condensation, and cell clumping, were observed in MCF-7 cells treated with
AgNPs, whereas control cells remained active. In addition, apoptotic features, such as cell
shrinkage and nuclear condensation and fragmentation, were also observed in MCF-7 tumor cells
48 h after treatment with 20 μg/mL of AgNPs. AgNPs synthesized with S. grandiflora extracts
induced the generation of free radicals, which resulted in oxidative damage and caspase-
mediated apoptosis [95].
AgNPs synthesized with Guignardia mangiferae extracts exhibited potent antifungal
activity against plant pathogenic fungi. IC50 values of AgNPs were 63.37, 27.54, and 23.84
μg/mL against normal African monkey kidney (Vero), HeLa (cervical), and MCF-7 (breast)
cells, respectively, after a 24-h incubation period. Thus, AgNPs synthesized with G. mangiferae
extracts are highly biocompatible, have potentially wider applicability, and should be explored as
promising candidates for a variety of biomedical/pharmaceutical and agricultural applications
[96]. AgNPs were synthesized using extracts from different plant origins:
Cucurbita maxima (petals), Moringa oleifera (leaves), and Acorus calamus (rhizome).
Among the three synthesized nanoparticles, AgNPs synthesized with A. calamus rhizome
extracts had enhanced antimicrobial and anticancer activity, which were evaluated through MTT
assays against epidermoid A431 carcinoma cells.
AgNPs synthesized with A. calamus rhizome extracts were superior to AgNPs generated
with petal and leaf extracts in their antimicrobial and anticancer activities [97]. Both treated
(synthesized) and untreated AgNPs induced DNA fragmentation at all concentrations [98].
Compared to untreated cells, cells treated with AgNPs synthesized using Phytolacca decandra,
Hydrastis canadensis, Gelsemium sempervirens, and Thuja occidentalis extracts exhibited DNA
laddering, confirming the apoptotic effects of nanoparticles. Specifically, AgNPs synthesized
using P. decandra and G. sempervirens extracts effectively induced DNA laddering compared to
AgNPs synthesized using H. canadensis and T. occidentalis extracts [98]. The IC50 values of
AgNPs synthesized using Potentilla fulgens extracts were 4.91 and 8.23 μg/mL in MCF-7 and U-
87 cell lines, respectively.
Furthermore, the cytotoxic effects of nanoparticles were evaluated against cancerous and
normal cells using trypan blue assay and flow cytometric analysis. In contrast to their effect on
normal cells, nanoparticles are capable of impairing or killing cancerous cells [29]. AgNPs
synthesized with Coleus amboinicus extracts were cytotoxic to EAC cell lines. AgNPs induced
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50 and 70 % cytotoxicity at 30 and 50 μg/mL, respectively, indicating concentrationdependent

cytotoxicity [99]. AgNPs synthesized with alcoholic flower extracts of Nyctanthes arbor-tristis
can be used for molecular imaging and drug delivery. Even at the highest concentration tested
(250 μg/mL), AgNPs were only marginally toxic to L929 cells [100]. The anticancer activity of
AgNPs synthesized with unripe fruits of Solanum trilobatum against a human breast cancer cell
line (MCF-7) was evaluated in vitro using MTT assays, nuclear morphological characteristics,
and RT-PCR and western blot analyses. MCF-7 cells treated with either AgNPs or cisplatin
exhibited decreased Bcl-2 expression and increased Bax expression, indicating the involvement
of mitochondria in the mechanism of death induced by AgNPs [101].
Mitochondria function as critical centers of signaling; their integrity can be
compromised by various regulators of apoptosis [102, 103]. The generation of ROS by AgNPs
may also require mitochondria, which may initiate intrinsic caspase-dependent apoptotic
pathways leading to cell death. Nanoparticles synthesized with Rosa indica extracts have the
potential to be used in a wide range of therapeutic anticancer applications. AgNPs synthesized
with green petals of R. indica act as radical scavengers and induce apoptosis in HCT-15 cells and
the generation of ROS [104].
ANTIOXIDANT ACTIVITY OF AGNPS

AgNPs synthesized using Leptadenia reticulata leaf extracts, at a concentration of 500
μg/mL, have the highest recorded radical scavenging activity of 64.81 % [105]. Plant extracts
promote DPPH radical scavenging activity of AgNPs, which is dose dependent. The abilityof
antioxidants to scavenge DPPH radicals is likely due to their ability to donate hydrogens and
easily incorporate electrons; the latter is possible due to the presence of host lipophilic radicals.
A change of color from purple to yellow was observed at 517 nm [106]. The DPPH radical
scavenging activity of HAuCl4 and AgNO3 was trivial compared to nanoparticles, which may be
due to salt conditions or weaker solubility of metal the oxides [107]
A sophisticated reaction can be observed between phenolic compounds and
phosphotungstic and phosphor molybdic acids in Folin-Ciocalteu reagents [108]. Phenolic
compounds present in plant extracts exhibited high antioxidant and reduction activities, which
are important for the synthesis of AgNPs [109]. The higher total phenolic content of Eclipta
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prostrata leaf extracts supports the assembly of silver ions into smaller AgNPs, because of the
donation of electrons by these compounds [110].
ANTIDIABETIC ACTIVITY OF AGNPS
The ability of AgNPs synthesized using stem extracts of Tephrosia tinctoria to control
blood sugar levels was evaluated. AgNPs scavenged free radicals, decreased levels of enzymes
that catalyze the hydrolysis of complex carbohydrates (α-glucosidase and α-amylase), and
increased the consumption rate of glucose [111]. Due to the adverse effects of methylene blue
(MB) on the environment, the removal of MB from wastewater is an important area of research
and a key challenge for researchers. AgNPs synthesized with aqueous stem extracts of Salvadora
persica were able to degrade MB in a light-dependent manner; by converting hazardous materials
into nonhazardous ones, AgNPs potentially have significant applications in water purification
[112]. MB exhibits characteristic absorption peaks at 663 and 614 nm, which were used to
monitor the photodegradation of MB. Higher concentrations induce the aggregation of AgNPs,
leading to an increase in particle size and a decrease in specific surface area and surface active
sites of particles [113]. The most effective concentration of AgNPs for the photo-degradation of
MB was 8 mg.
DIFFERENT FIELD APPLICATIONS OF AGNPS

Nanotechnology applications are highly suited for biological molecules because of
their unique properties. Nanotechnology is a growing area of research in the fields of material
science and biological science [114]. Silver nanoparticles have attracted the attention of
researchers because of their broad applications in diverse areas, such as integrated circuits [115],
sensors [116], biolabeling, filters, antimicrobial deodorant fibers [117], cell electrodes [43], low-
cost paper batteries (silver nano-wires) [118], and antimicrobials [119]. AgNPs have been used
extensively as antimicrobial agents in
The healthy industry, food storage, textile coatings, and a number of environmental applications
[120], few of which are shown in Fig. 4. Antimicrobial properties of AgNPs are beneficial for
different fields of medicine, various industries, animal husbandry, packaging, accessories,
cosmetics, health, and the military. In general, therapeutic effects of silver particles (in
suspension) depend on different parameters, including particle size (surface area and energy),
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particle shape (catalytic activity), particleconcentration (therapeutic index), and particle charge
(oligodynamic quality) [121].
The viability of A549 cells treated for 6 h with 10 and 50 μg/mL of AgNPs
synthesized with Albizia adianthifolia leaf extracts was 21 and 73 %, respectively, and that of
normal peripheral lymphocytes was 117 and 109 %, respectively, indicating that AgNPs are
nontoxic to normal PLs cells [122].
Fig. 3 Plant-mediated synthesis of silver nanoparticles: their characteristic properties and

therapeutic applications
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Fig. 4 Different applications of synthesized silver nanoparticle

AgNPs synthesized with Indigofera aspalathoides extracts were tested in wound
healing applications following excision in animal models [28]. AgNPs synthesized with
Chrysanthemum morifolium extracts were added to clinical ultrasound gels, which are used with
an ultrasound probe, and were found to have bactericidal activity, contributing to the sterility of
the instrument [123]. AgNPs synthesized with M. zapota leaf extracts exhibited acaricidal
activity against Rhipicephalus (Boophilus) microplus (LC50 = 3.44 mg/L) [58]. The IC50 values
of AgNPs synthesized using aqueous extracts of Ashoka or neem leaves against Plasmodium
falciparum were 8 and 30 μg/mL, respectively [124]. Appreciable larvicidal activity of AgNPs
synthesized with aqueous extracts of E. prostrata was observed against Anopheles subpictus and
Culex tritaeniorhynchus [125]. The population of bacteria decreased after 6 h when 10 mg of
AgNPs synthesized using P. juliflora leaf extracts was used to treat 100 mL of sewage and
increased over time [56]. AgNPs synthesized with Acacia nilotica pod extracts were used to treat
glassy carbon electrodes, which exhibited greater catalytic activity in reducing benzyl chloride
than glassy carbon and metallic Ag electrodes [126]. AgNPs synthesized using Gloriosa superba
extracts act through the electron relay effect and influence the degradation of MB after 30 min
[127].
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CONCLUSIONS:
In summary, silver nanoparticles (AgNPs) exhibit remarkable physical, mixture,
optical, and natural properties compared to other biomedical nanomaterials, which make them
ideal in various stages of diverse biomedical applications. Nanoparticles synthesized with plant
concentrates have yielded promising results in biomedical applications. Comprehensive
examination further contemplated that repercussions of nanoparticles give essentially the same
number of preferences and purposes of enthusiasm for remedial applications in examination to
standard medicines and antidotes to poisons. The use of AgNPs in medicinal transport systems
may also be utilized in the future in the field of arrangement. AgNPs have the potential to
function as therapeutics with diverse clinical and pharmacological properties.
They may be used in broad applications, including as anticancer agents or bactericidal
agents during surgery or recovery. In this way, the green synthesis of AgNPs as novel remedial
authorities will be significant in various biomedical applications. Notwithstanding their potential
in restorative applications, the impact of AgNPs on human welfare (both positive and negative)
should be completely considered before their widespread use. The adaptability of manufacturing
techniques for AgNPs and their easy reconstitution into distinct media have prompted further
research into the hypothetical impact of nanoparticles as antimicrobial, antiviral, and mitigating
agents. The shape, size, and size distribution of AgNPs can be controlled by modifying synthesis
conditions, such as with specialists, stabilizers, or distinct engineering techniques. The
productive translation of silver into nanotechnology applications requires safe, creative, and eco-
conscious strategies and greater control over their biodistribution and pharmacokinetics in
clinical applications.
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of silver nanoballs against E. Coli MTCC 1302, S.typhimurium MTCC 1254, B. subtilis
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127. Jebakumar TN, Edison I, Sethuraman MG (2013) Electrocatalytic reduction of

benzyl chloride by green synthesized silver nanoparticles using pod extract of Acacia
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nanoparticles from Gloriosa superba L. leaf extract and their catalytic activity.
129. Sana SS, Badineni VR, Arla SK, Boya VKN (2015) Eco-friendly synthesis of
silver nanoparticles using leaf extract of Grewia flaviscences and study of their
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131. Mishra PM, Sahoo SK, Naik GK, Parida N (2015) Biomimetic synthesis,
characterization and mechanism of formation of stable silver nanoparticles using
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of Carica papaya leaf extract coated silver nanoparticles through X-ray diffraction,
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136. Murugan K, Dinesh D, Kumar PJ et al (2015) Datura metal-synthesized silver
nanoparticles magnify predation of dragonfly nymphs against the malaria vector
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137. Swamy MK, Akhtar MS, Mohanty SK, Sinniah UR (2015) Synthesis and
characterization of silver nanoparticles using fruit extract of Momordica cymbalaria and
assessment of their in vitro antimicrobial, antioxidant and cytotoxicity activities.
138. Roni M, Murugan K, Panneerselvam C, Subramaniam J, Nicoletti M et al (2015)
139. Characterization and biotoxicity of Hypnea musciformis-synthesized silver
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xylostella. Ecotoxicol Environ Saf 121:31–38
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extractmediated silver nanoparticles synthesis and its larvicidal potential against dengue,
malaria and filariasis vector. Parasitol Res 114(8):3087–3096 Chung et al. Nanoscale
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nanoparticles using Vasaka (Justicia adhatoda L.) leaf extract. Indian J Microbiol
55(2):163–167
142. Latha M, Sumathi M, Manikandan R, Arumugam A, Prabhu NM (2015)
Biocatalytic and antibacterial visualization of green synthesized silver nanoparticles
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antibacterial activity of silver nanoparticles using Emblica officinalis fruit extract.
144. Korbekandi H, Chitsazi MR, Asghari G, Bahri Najafi R, Badii A, Iravani S (2015)
Green biosynthesis of silver nanoparticles using Quercus brantii (oak) leaves
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145. Bhakya S, Muthukrishnan S, Sukumaran M, Muthukumar M (2015) Biogenic
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146. Perugu S, Nagati V, Bhanoori M (2015) Green synthesis of silver nanoparticles
using leaf extract of medicinally potent plant Saraca indica: a novel study. Appl Nanosci.
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147. Prathap M, Alagesan A, Ranjitha Kumari BD (2014) Anti-bacterial activities of

silver nanoparticles synthesized from plant leaf extract of Abutilon indicum (L.) wweet. J
Nanostruct Chem 4:106
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biosynthesis of silver nanoparticles using Prosopis farcta extract and its antibacterial
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149. Chitra G, Balasubramani G, Ramkumar R, Sowmiya R, Perumal P (2015) Mukia
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150. Borase HP, Salunkhe RB, Patil CD, Suryawanshi RK, Salunke BK, Wagh ND,
Patil SV (2015) Innovative approach for urease inhibition by Ficus carica extract-
fabricated silver nanoparticles: an in vitro study. Biotechnol Appl Biochem.
doi:10.1002/bab.1341
151. Khatami M, Pourseyedi S, Khatami M, Hamidi H, Zaeifi M, Soltani L (2015)
Synthesis of silver nanoparticles using seed exudates of Sinapis arvensis as a novel
bioresource, and evaluation of their antifungal activity. Bioresources and Bioprocessing
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152. Rajkuberan C, Sudha K, Sathishkumar G, Sivaramakrishnan S (2015)
Antibacterial and cytotoxic potential of silver nanoparticles synthesized using latex of
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153. Sreekanth TV, Ravikumar S, Eom IY (2014) Green synthesized silver
nanoparticles using Nelumbo nucifera root extract for efficient protein binding,
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154. Joseph S, Mathew B (2014) Microwave assisted facile green synthesis of silver
and gold nanocatalysts using the leaf extract of Aerva lanata. Spectrochim Acta A Mol
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155. Zuas O, Hamim N, Sampora Y (2014) Bio-synthesis of silver nanoparticles using
water extract of Myrmecodia pendan (sarang semut plant). Mater Lett 123:156–159
156. Yousefzadi M, Rahimi Z, Ghafori V (2014) The green synthesis, characterization
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157. and antimicrobial activities of silver nanoparticles synthesized from green alga
Enteromorpha flexuosa (wulfen). J Agardh Mater Lett 137:1–4
158. Shankar S, Jaiswal L, Aparna RSL, Prasad RGSV (2014) Synthesis,
characterization, in vitro biocompatibility, and antimicrobial activity of gold, silver and
gold silver alloy nanoparticles prepared from Lansium domesticum fruit peel extract.
Mater Lett 137:75–78
159. Ghaffari-Moghaddam M, Hadi-Dabanlou R (2014) Plant mediated green
synthesis and antibacterial activity of silver nanoparticles using Crataegus douglasii fruit
extract. J Indust & Eng Chem 20:739–744
160. Shetty P, Supraja N, Garud M, Prasad TNVKV (2014) Synthesis, characterization
and antimicrobial activity of Alstonia scholaris bark-extractmediated silver nanoparticles.
J Nanostruct Chem 4:161–170
161. Maiti S, Krishnan D, Barman G, Ghosh SK, Laha JK (2014) Antimicrobial
162. activities of silver nanoparticles synthesized from Lycopersicon esculentum
extract. J Analy Sci & Tech 5:40
163. Banerjee P, Satapathy M, Mukhopahayay A, Das P (2014) Leaf extract mediated
green synthesis of silver nanoparticles from widely available Indian plants: synthesis,
characterization, antimicrobial property and toxicity analysis. Biores & Bioproces 1:3
164. Roopan SM, Madhumitha G, Abdul Rahuman A, Kamaraj C, Bharathi A,
Surendra TV (2013) Low-cost and eco-friendly phyto-synthesis of silver nanoparticles
using Cocos nucifera coir extract and its larvicidal activity. Ind Crop Prod 43:631–635
165. Suman TY, Radhika Rajasree SR, Kanchana A, Elizabeth SB (2013)
Biosynthesis, characterization and cytotoxic effect of plant mediated silver nanoparticles
using Morinda citrifolia root extract. Colloids Surf B: Biointerfaces 1(106):74–78.
166. Das S, Das J, Samadder A, Bhattacharyya SS, Das D, Khuda-Bukhsh AR (2013).
Biosynthesized silver nanoparticles by ethanolic extracts of Phytolacca decandra,
Gelsemium sempervirens, Hydrastis canadensis and Thuja occidentalis induce
differential cytotoxicity through G2/M arrest in A375 cells. Colloids Surf B Biointerfaces
101:325-36.
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MACHINE LEARNING IN DRUG REPURPOSING

Kiruthika G and Dr Vasna Joshua
National Institute of Epidemiology (ICMR), Chennai
kiruthikabiostat@gmail.com
Drug repurposing means repurposing or re-profiling already approved or investigational

drugs to treat common and rare diseases. During the time of a pandemic, drug repurposing serves
as a very effective alternative to drug development as it cuts down both time and expenses
incurred in a drug developmental process. One such famous repurposed drug is thalidomide
which was first used as an over-the-counter sedative for morning sickness but later it is being
used to treat leprosy and multiple myeloma1. Drug repositioning primarily based on the concept
of molecular similarity which assumes similar target proteins share similar binding
characteristics to compounds4. In the recent years machine learning is commonly used to
repurpose drugs. System biology approaches like sequence and structural similarities among
drug targets, side-effect similarity and disease-drug networks have been used by analyzing large
datasets like gene expression profiles.
Kim et al used machine learning techniques like logistic regression, random forest and
support vector machine (SVM) as classification algorithms for predicting the potential
indications (unknown pharmacological effects) of existing drugs and herbal compounds 2. Jeon
et al used SVM-recursive feature elimination (SVM-REF) for feature selection and SVM with
radial basis function (RBF) to classify proteins as cancer and non-cancer drug targets 3. A 10-fold
cross-validation was performed to validate the classification. SVM, a supervised learning
algorithm is widely used for drug repurposing mainly for two reasons. Firstly, it can handle large
and noisy datasets. Secondly, SVM is robust to over-training.
REFERENCES
1. Ashburn, T., Thor, K. Drug repositioning: identifying and developing new uses for
existing drugs. Nat Rev Drug Discov 3, 673–683 (2004). https://doi.org/10.1038/nrd1468
2. Kim et al. BMC Bioinformatics 2019, 20(Suppl 10):247
3. Jeon et al. Genome Medicine 2014, 6:57
4. Schaduangratet al. J Cheminform (2020) 12:9
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WOMEN’S HEALTH
Salma Suqlain M
B.Sc. Biotechnology.
Marudhar Kesari Jain College for Women, Vaniyambadi
salmasuqlain096@gmail.com
Being a man or woman has a significant impact on health, As a result on both biological
and gender related differences the health of woman and girls is of particular concern because in
many societies, they are disadvantaged by discrimination rooted in socio-cultural factors
example: WOMEN AND GIRLS FACE INCREASED VULNERABILITY TO HIV\AIDS.
While poverty is an important barrier to positive health outcome for both men and women,
poverty tends to yield a higher burden on women and girls health due to for eg: FEEDING
PRACTICES (malnutrition) and use UNSAFE COOKING FUELS.
Some health issues effect women differently and more commonly .Many women’s
health condition goes undiagnosed and most drug trials do not include female test subjects.
Even, so women bear exclusive health concerns, such as BREAST CANCER,
CERVICAL CANCER, MENOPAUSE and PREGNANCY. Women suffer higher heart attacks
compared to men .Depression and anxiety exhibit more frequently among female patients.
URINARY TRACT INFECTIONS present more often in females and SEXUALLY
TRANSMITTED DISEASES can cause more harm to women.
BREAST CANCER:
This initiates in the lining of the milk ducts, it can spread to other organs and is most
aggressive cancer affecting the global female population. Initially, women afflicted with breast
lumps. Most breast lumps are not threatening, but it is important for women to check each one by
a care provider.
OVARIAN AND CERVICAL CANCER:
Many people are not aware of the differences between OVARIAN & CERVICAL
CANCER. Cervical cancer occurs from the lower uterus, while ovarian cancer starts from
Fallopian tubes. Both cause similar pain . Cervical cancer also causes DISCHARGE & PAIN
during INTERCOURSE.
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GYNECOLOGICAL HEALTH:
Bleeding and discharge are normal part of menstrual cycle. However, added symptoms
during menstruation may indicate health issues , such as bleeding between menstruation &
frequent urinating can mimic other conditions. VAGINAL ISSUES could also indicate serious
problems such as SEXUALLY TRANSMITTED DISEASES or reproductive tract cancer.
PREGNANCY ISSUES:
Pre-existing diseases can worsen during pregnancy, threatening the health of a mother or
her child. DEPRESSION CAN HARM THE MOTHER AND CHILD DURING PREGNANCY.
If not managed properly. Pregnancy can cause the healthy mother’s RBC count to drop, a
condition called ANEMIA or INDUCE DEPRESSION.
ISBN: 978-81-948129-1-3 Page 481

DNA FIX
Shubham
Dyal Singh College , Karnal (132001) , Haryana , India
Mobile no.-8708075152
bhattishubham25@gmail.com
Life forms are for all time presented to endogenous and exogenous operators that harm
DNA. If not fixed, such harm can bring about changes, sicknesses and cell demise. The cell
reactions to DNA harm incorporate cycles that manage its outcomes (for example resilience and
apoptosis) just as immediate adjustment of the harm by DNA fix systems, which may require
enactment of checkpoint pathways. There are different types of DNA harm, for example, base
alterations, strand breaks, crosslinks and confuses. There are likewise various DNA fix
pathways. Each fix pathway is coordinated to explicit kinds of harm, and a given sort of harm
can be focused by a few pathways. Significant DNA fix pathways are bungle fix (MMR),
nucleotide extraction fix (NER), base extraction fix (BER), homologous recombinational fix
(HR), and non-homologous end joining (NHEJ). These pathways each require various proteins.
On the other hand, O-alkylated bases, for example, O6-methylguanine can be fixed by the
activity of a solitary protein, O6-methylguanine-DNA methyltransferase (MGMT).
MGMT eliminates the alkyl bunch in a self destruction response by move to one of its
cysteine buildups. Photolyases can part covalent obligations of pyrimidine dimers created by UV
radiation. They tie to an UV injury in a light-free cycle, yet require light (350-450 nm) as a fuel
hotspot for fix.
Another NER-free pathway that can eliminate UV-incited harm, UVER, is available in
just a couple of life forms, for example, the yeast Schizosaccharomycespombe. A key factor in
UVER is the endonuclease Uve1/UVDE, which cuts 5′ of different kinds of harm. Late work has
revealed novel pathways, for example, record coupled BER, break-actuated replication, and
nucleotide entry point fix just as interconnections between known pathways. For
straightforwardness, we don't consider these here. Albeit most fix proteins are normally
homologous between livings beings, their assignments are regularly extraordinary. Here we by
and large utilize the names of human proteins.
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MISMATCH REPAIR
The principle assignment of MMR is to eliminate base jumbles and little addition/erasure
circles (IDLs) presented during replication. In Escherichia coli, the fundamental parts in MMR
are MutS, MutL and MutH. MutH scratches the non-methylated strand and along these lines
empowers segregation between the recently integrated strand and the layout. MMR is
bidirectional, for example scratching and debasement can happen from either the 5′ or 3′ side of
the confuse.
In eukaryotes, a few MutS and MutL homologues are engaged with MMR; MutH
homologues seem, by all accounts, to be missing. Inactivation of human MMR causes inherited
nonpolyposis colorectal disease (HNPCC) and a few kinds of inconsistent tumor. Throughout
human MMR, base confounds are limited by the MutS-homologous heterodimer MSH2-MSH6,
while little IDLs can be limited by MSH2-MSH6 and MSH2-MSH3. Consequently, the MutL-
homologous heterodimer MLH1-PMS2 is selected. In certain eukaryotes extra MutLhomologues
exist.
These structure heterodimers with MLH1 and may assume a minor function in MMR. It
isn't yet seen how eukaryotes recognize the new and the old strand. Strand separation might be
either intervened by the replication adornment factor PCNA or could be essentially accomplished
by acknowledgment of scratches, holes or free 3′ closes that are available in the early strand
during replication. In a downstream advance, the recently combined strand is corrupted, which
eliminates the bungle. MMR patches are ∼100 to >1000 nucleotides long. EXO1 is associated
with 5′ to 3′ extraction. It isn't yet clear which components take an interest in 3′ to 5′ extraction,
yet DNA Pol δ and ϵ and EXO1 might be included. MMR is finished after DNA amalgamation
by the replication apparatus and ligation of the rest of the scratch.
NUCLEOTIDE EXTRACTION FIX
NER eliminates an assortment of types of DNA harm, including photoproducts initiated
by UV and other massive injuries. NER comprises of two subpathways: worldwide genome fix
(GGR), which eliminates harm in the genome generally speaking and record coupled fix (TCR),
which explicitly fixes the interpreted strand of dynamic qualities. The principle contrast among
GGR and TCR is the necessity for various elements during the underlying acknowledgment
steps. UV-DDB, comprising of DDB1 and DDB2, and XPC-hHR23B are engaged with the
ISBN: 978-81-948129-1-3 Page 483

acknowledgment step of GGR, while TCR is believed to be started by RNA polymerase II

slowed down at an injury. Extra factors required for TCR are CSA and CSB.
The proteins acting further downstream in GGR and TCR are probably going to be
indistinguishable. To begin with, record factor IIH (TFIIH), a complex comprising of nine
subunits, is enlisted to the harmed site. At this progression the underlying acknowledgment
factors are presumably delivered from the harmed DNA. Two subunits of TFIIH, XPB and XPD,
display helicase movement of inverse extremity, and loosen up the DNA around the injury. The
following variables that predicament to the harmed site are XPG and XPA-RPA. XPA-RPA
checks whether the NER complex is accurately collected and guarantees legitimate cut of the
harmed strand. In the wake of official of XPF-ERCC1, double entry point happens by XPG and
XPF-ERCC1, which slice 3′ and 5′ to the harm, individually. Along these lines, the harm is
delivered in a 24-32 nucleotide long oligonucleotide. Fix is finished by DNA combination and
ligation. The ordinary problem brought about by an imperfection in NER is
xerodermapigmentosum (XP), while Cockayne disorder (CS) and trichothiodystrophy (TTD) are
because of disabled TCR and in the last case inevitably additionally to influenced record.
BASE EXTRACTION FIX
BER basically fixes non-cumbersome sores created by alkylation, oxidation or
deamination of bases. Cells contain a few DNA glycosylases, every one of them displaying a
particular substrate range. After cleavage of the N-glycosylic bond by a DNA glycosylase, the
harmed base is delivered and an apurinic/apyrimidinic (AP site) is made. An AP site can likewise
happen unexpectedly and speaks to harm itself.
Bifunctional glycosylases have an inborn AP lyase action, which severs the sugar-
phosphate spine 3′ to the AP site. The subsequent divided sugar buildup is eliminated by a
phosphodiesterase action, contributed by either an AP endonuclease or by DNA polymerase β.
The one-nucleotide hole is filled by Pol β and ligated. Handling of AP destinations created by a
monofunctional DNA glycosylase requires 5′ cut by an AP endonuclease (the significant human
AP endonuclease is APE1). Pol β joins a nucleotide and its deoxyribophosphodiesterase
(dRPase) action eliminates the 5′ moiety. The rest of the scratch is fixed by ligation. During a
minor, long-fix BER pathway, 2-8 nucleotides are taken out along with the harmed nucleotide.
Long-fix BER might be required within the sight of changed AP destinations where the
5′ moiety can't be taken out by a dRPase action. After strand removal by Pol β, and Pol δ or Pol
ISBN: 978-81-948129-1-3 Page 484
ϵ, a fold structure is shaped, which is severed by FEN1. No human ailment is as of now known to
be related with an imperfection in BER, which might be because of undeveloped lethality or
practical excess as well as on the grounds that collection of harm, generally prepared by BER,
has no natural outcome. Truth be told, knockout mice lacking components acting downstream of
DNA glycosylases show an early stage deadly aggregate, while an imperfection in a solitary
DNA glycosylase doesn't bring about any phenotypic anomaly.
HOMOLOGOUS RECOMBINATION FIX
Twofold strand breaks (DSBs) can be fixed by either HR or NHEJ. HR utilizes a
homologous DNA layout and is exceptionally precise, though NHEJ rejoins the messed up closes
without utilizing a format and is frequently joined by loss of certain nucleotides.
The general commitment of every pathway relies upon the phone cycle stage, with NHEJ
being more dynamic in G1 and HR overwhelming during S and G2 stages. During HR DSBs are
changed over to 3′ single-abandoned DNA (ssDNA) tails, which are limited by RPA. Handling
of DSBs most likely requires MRE11-RAD50-NBS1. RAD52 communicates with RPA and
elevates official of RAD51 to the ssDNA, which might be balanced out by RAD51 paralogues
(RAD51B, RAD51C, RAD51D, XRCC2 and XRCC3 in human, RAD55 and RAD57 in yeast).
Accordingly, the RAD51-bound ssDNA attacks a homologous atom in a response
invigorated by RAD54. After DNA blend and ligation, two Holliday intersections are shaped and
branch movement can happen. The Holliday intersections are at last settled by resolvases, which
in eukaryotes are not yet recognized.
HR likewise speaks to a mistake free subpathway of harm resistance, permitting
replicational sidestep of sores through a layout switch. Then again, harm resilience can be
accomplished by mistake free and blunder inclined translesion amalgamation completed by
particular DNA polymerases. HR-subordinate injury sidestep may in some cases produce a 3′
fold that can be severed by MUS81-EME1 or settled by TOP3-RECQ.
NON-HOMOLOGOUS END JOINING
NHEJ is started by official of Ku70-Ku80 dimers to the DNA closes. In higher
eukaryotes the DNA protein kinase synergist subunit (DNA-PKcs) is in this way enrolled. DSBs
that are not appropriate for ligation might be handled by MRE11-RAD50-NBS1 and different
nucleases, for example, FEN1.
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Moreover, a DNA polymerase might be required. At long last, the DNA closes are
rejoined by XRCC4-DNA ligase IV.
Deficient fix of DSBs can bring about chromosomal precariousness, which is portrayed
by revisions and loss of chromosomes. Various human conditions, for example, Ataxia
telangiectasia (AT) and related issues, Nijmegen breakage disorder (NBS), just as bosom and
ovarian malignant growth brought about by transformation of BRCA1 or BRCA2, are related
with surrenders in DSB fix. Notwithstanding, these conditions are a result of imperfections in
guideline of DSB fix (for example in checkpoint enactment) as opposed to because of an
immediate inactivation of HR or NHEJ.
DEPRESSION AND ANXIETY:
Natural hormonal fluctuations can lead to depression or anxiety. PMS (premenstrual
syndrome) occurs in women , while premenstrual dysmorphic disorder (PMDD) presents similar
but greatly intensified symptoms. Shortly after birth of baby, many mothers acquire a form of
depression called “baby blues”.
PERIODS PROBLEMS:
Regular periods are sign that a body is working normally. One should have regular period
unless your pregnant, breast feeding, postmenopausal, or have a medical condition that causes
your periods to stop . Irregular heavy or painful periods are signs of a serious health problem.
Irregular period can also make it harder to get pregnant.
BASIC TIPS ON HEALTHY LIVING FOR WOMEN:
1. Take 5 supplements multivitamin, fish oil, probiotics, vitamin D, magnesium.
2. Eat to 80% full.
3. Eat clean not processed whole food, organic fruits and vegetables, meat and dairy.
4. Include exercise in the daily routine.
5. Manage stress.
6. Sleep- this is the foundation for almost everything.
7. Practice self-care.
8. Maintain weight.
9. Keep your body hydrated.
10. Stay active.
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VULVAR CARE:
1. Keep an eye on your discharge- unusual colours and consistencies can signal a problem.
2. Its normal for vagina to have a slight scent, but when it becomes too strong or smells off,
one may need medical attention.
3. If your experiencing vaginal itching and burning, don’t just assume it’s a yeast infection.
4. Don’t use products like petroleum jelly or oil for lube.
5. If you see blood clots bigger than a quarter during your periods, talk to your doctor.
6. Use plain unperformed soap to vagina.
7. Maintain safer sex to keep harmful germs away.
8. Cervical screening-all women aged from 25-64 should be screened.
9. Protect vaginal Ph balance without douching.
10. Keep the undergarments dry.
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“A WOMAN’S HEALTH IS HER CAPITAL”

THE RELATIONSHIP BETWEEN SYMPTOM BURDEN AND SYSTEMIC
INFLAMMATION DIFFERS BETWEEN MALE AND FEMALE ATHLETES
FOLLOWING CONCUSSION.
Sowmiya. N,
III B Tech in Biotechnology, PSG College of Technology, Peelamedu, Coimbatore,
Tamil Nadu.
INTRODUCTION:
To guide the patients in the recovery process, who are involved in concussion mainly
those who are participating in the sports (Sport-Related Concussion) including both male and
female, symptom burden plays a main role for the doctors for the resolution of such symptoms.
Whereas inflammatory response also involves in concussion pathophysiology, here comes a
great confusion regarding how to differentiate between these two terminologies and responses.
The main purpose to review this article is to get clear cut idea about the differences and the
relations between the symptom burden and systemic inflammation through the biomarkers and
the blood samples testing in both male and female gender.
CONTENT:
• Symptom burden act as a tool for the curative process in the resolution of
symptoms which is a prerequisite for medical clearance and helps in recovery.
• The systemic inflammation is characterised by the presence of symptoms in
which various inflammatory mediators were linked and altered blood concentrations of
inflammatory cytokines and chemokines.
• These mediators can communicate with the primary afferent and secondary
efferent in the CNS to respond those cytokines.
• The range of inflammatory mediators the evaluates the activity of the symptoms
and inflammation.
• Here, data were collected from both men and women with no significant
difference in the in the clinical characteristics
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• The concentrations of many biomarkers preferred shows no significant difference

once the corrections made in the concentration of the biomarkers used till that it shows either
lower in females or higher in females when compared to males.
• After many analyses, symptom severity shows inverse action with inflammatory
cytokines in the peripheral blood of female athletes than male even though the action of
symptom burden and its recovery of time is same for both male and female.
• This shows that greater symptom burden was associates with the lower
concentrations of the inflammatory cytokine’s interferon-gamma and Tumour Necrosis Factor-
alpha and the innate immune function marker MPO in female athletes of SRC.
• This shows a positive relationship with the chemokine MCP-4 used.
• In female, menstrual cycle had an effect which changes in the immune function
due to hormonal changes during the menses because it depends on the blood concentration and
blood biomarker.
• These made in males a positive symptom correlated with blood concentration of
IFN-gamma following SRC.
• More than twenty biomarkers were quantitated by the immunoassay.
• To evaluate the symptoms, Sport Concussion Assessment Tool Version-5(SCAT-
5) is used.
• It also includes the clear study about the neuroimaging and blood biomarkers
because these studies still remain unclear.
CONCLUSION:
The unique inflammatory data in male and female gives the system burden following
SRC. The symptoms severity is correlated with the elevated blood concentrations of different
biomarkers like Interferon and TNF-alpha and the MPO in females and these are more
generalised in the males when compared to females. So, there is slight difference and the
inference in the data of the sex gender it is required for the future studies with the statically
analysis for association between the symptom reporting and the inflammatory biomarkers
following SRC with and without potential compounds with statically analytical design.
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A REVIEW ON THE APPLICATION OF MICROBES IN FORENSICS

Haritha S
II BSc BIOTECHNOLOGY
PSG COLLEGE OF ARTS & SCIENCE, COIMBATORE,
divhari23@gmail.com
ABSTRACT
The significant technological advances in the 21st century was a boon to forensic science
ranging from DNA fingerprinting to MPS and metagenomics and provided investigations with
enough evidence to produce in court and to convict criminals. The fact that the microbiome
hosted by every individuals unique like a personal signature serves as an impetus to the forensics
field and can be exploited to gain investigative leads by sequencing the genome of those
microorganisms and obtaining an overview of the microbial community pattern. Frequently used
evidence like the skin, hair and body fluids have their own microbe composition which helps in
individualization and can help overcome limitations like non- availability or availability of low
quality human DNA. The PMI estimation is quite unreliable in terms of using maggots and
insects but its accuracy can be improved by employing the microbial clock which can help
determine the stage of decomposition of the body depending on the microbe community present
at that time frame. The only drawback to applying microbes in forensics is the lack of reference
database and microbial genome sequences.
KEYWORDS: Metagenomics, DNA, Fingerprinting.
INTRODUCTION
The human body harbours 500–1000 different species of bacteria and each strain has a
genome containing thousands of genes and it is clear that the total DNA of microbes inhabiting
our bodies,the microbiome, offers much more genetic diversity than the human genome[Sarah
Gino and Luciana Caenazzo ,Microorganisms ;8(6), 9 June 2020].Humans shed ~30 million
bacterial cells into their vicinity every hour (Qian et al., 2012).The fluctuations in the structure
and composition of the microbiome may contain useful information like host lifestyle, diet,
occupation, travel, and pharmaceutical use that has huge forensic potential. Microbiome profiling
can potentially complement DNA profiling but a substantial amount of research is still required
to prove that residual microbial fingerprints can be used as effective trace evidence [Hampton-
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Marcell, Jarrad.T et al., Microbial biotechnology vol. 10,2 ; March 2017].Another aspect of
forensics which is difficult to establish is PMI(Post Mortem Interval) because of relatively poor
understanding of corpse decomposition but using microbial community change it is possible to
track the progression of decomposition because microbes are ubiquitous in the environment and
located on humans before death, can be reliably quantified using high-throughput DNA
sequencing. So microbes can also be used as a ‘clock’ to estimate PMI [Metcalf JL,et al., Elife;2,
Oct 15 2013]. The various technologies like Massive Parallel Sequencing have accelerated
research in this area.
SKIN MICROBIOME
Skin is the largest organ of human body, colonized by several microorganisms which
are mostly harmless or even beneficial to the host (Grice EA. et al.,Nat Rev Microbiol. 9(8),
2011 Aug). These microorganismsthat cover the surface of the skin and reside deep in the hair
and glands can be easily dispersed and transferred to surfaces upon touching and acts as a
personal signature, establishing a link between individuals and the objects they touch (Fierer et
al. 2010; Lax et al. 2015; Meadow et al. 2014). Additionally, each body part constitutes a
specific niche, with a particular microbiome composition, dictated by several factors (e.g., skin
thickness and folds, densities of hair follicles and the presence of both eccrine and apocrine
glands).
For instance, if Propionibacteria and Staphylococcus species dominate sebaceous sites
and Corynebacteria moist sites, β-Proteobacteria and Flavobacteriales dominate dry sites. Some
of them persist on surfaces and are extremely resistant to environmental stress (Brooke et al.
2009; Kong and Segre 2012; Smith et al. 1996) and also the longitudinal stability of skin
microbial community is site dependant( Grice et al. 2019). However the stability of the personal
profile and changes induced by antifungal and antibiotic therapies remain a doubt (Langdon et al.
2016; Raymond et al. 2016).
HAIR MICROBIOME
Hair is the most commonly found evidence at crime scenes but majority lacks the
anagenic root (actively growing hair), essential for DNA extraction and STR profile analysis
(Tridico et al. 2014; Tridico et al. 2017).
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In a recent study it was observed that majority of hairs recovered in investigations

correspond to scalp hairs while pubic hairs are used in sexual assault cases. Microbiome analysis
allows not only the individualization but also discriminate between the two hair sources. The
study also revealed that cohabiting couples interchange microbiomes during sexual intercourse.
Additionally, the microbiome present on scalp hairs is prone to compositional oscillations than
pubic hairs (Tridico et al. 2014). Metagenomic analyses of hairs unsuitable for nuDNA profiling
may provide a microbial fingerprint to augment other forensic results such as mtDNA analyses
(Tridico, S.R et al. Investigative Genetics 5, 16 (Dec 2014).
BODY FLUID MICROBIOME
Body fluid identification is an important aspect in forensic science, as the ability to
identify body fluids such as blood,semen,menstrual blood,saliva, urine, sweat and vaginal fluid is
often the key in any criminal investigation(Harrison S. et al., Research and Reports in Forensic
Medical Science. 2016;6:11-23).Traditional methods can only be used to identify a single
biological fluid, and thus elimination of the presence of other biological fluids and mixtures in
forensic samples can be problematic. These body fluids provide DNA evidence (Virkler and
Lednev 2009) but high quality of that DNA is essential for mRNA and other studies. It has been
proposed that each body fluid type presents a specific composition of microorganisms that can be
used as bioindicators (Hanssen et al. 2017). For instance, saliva could be identified by detection
of specific bacteria, such as Veillonellaatypica,
Streptococcus mutans and Streptococcus salivarius; while vaginal fluid could be
identified by detection of specific bacteria, such as Lactobacillus crispatus and Lactobacillus
gasseri (Choi et al. 2014; Giampaoli et al. 2017).
METHODOLOGIES EMPLOYED
Microbial forensic evidence may reside in a wide range of samples matrices including
food, water, air filters, swab and swipes, soil, animal tissue, and clinical samples (e.g., tissue,
sputum, blood, stool, urine). Therefore, an analyst must have a variety of sample processing
methods available to address the demands of myriad possible samples and scenarios.
Previously to the advent of DNA-based techniques, in the late 1980s, the available tools
for microorganism detection and identification were restricted to phenotypic methods associated
with antigenic and/or antimicrobial resistance profiles. These methods only allow resolution at
the genus and/or species level (using culture-dependent techniques) (Oliveira et al. 2018).
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Afterwards, a paradigm shift was observed: the traditional phenotypic methods were
supplanted by nucleic acid-based molecular methods [e.g., DNA fingerprinting, whole genome
sequencing and microarray analysis] (Budowle et al., 2005; Pattnaik and Sekhar 2008). These
methods not only improve taxonomical resolution (identification to isolate/strain level), but also
decreased the turnaround time between sample collection and results (Pattnaik and Sekhar 2008).
Current detection methods in the microbial forensic workflow can range from real-time
PCR, microarray, genetic typing, whole-genome sequencing, MPS, shotgun metagenomics and
beyond. (Oliveira M, Amorim A, Appl Microbiol Biotechnol. 102(24), October 2018)
MPS (MASSIVELY PARALLEL SEQUENCING)
In the early 2000s, the advent of massively parallel sequencing (MPS)(which also has
been referred to as next-generation sequencing [NGS] and high-throughput sequencing [HTS]),
allow more sensitive and have higher throughput, which will make it more applicable in
microbial forensics.
Larger genomes can be fully sequenced in a couple weeks and small genomes, such as
a bacterial genome, can be fully sequenced within a few days. MPS provides greater coverage
across a genome with higher depth of coverage at each site for increased confidence in base
calls, and barcoding allows for multiplexing of samples (from a few to hundreds) in a single
sequencing run.
Using NGS to sequence total DNA extracts from any sample allows both the sequencing
of the whole genome of a given microorganism and the examination of whole communities of
microbes, with the possibility of rapidly and efficiently identifying all different bacterial taxa and
strains, thereby obtaining an overview of the resident microbial population (Tozzo, Pamela et al.
Microorganisms vol. 8,6 ,Jun 2020).MPS also has several advantages over the conventional
Sanger sequencing (Bano et al. 2015; Clarke et al. 2017; Schmedes et al. 2017). Full
characterization of bacterial or viral genomes can be achieved in a number of days as compared
to more limited traditional methods, such as culturing, MLST (multi-locus sequencing typing)
and PFGE (pulsed-field gel electrophoresis), which can take several days to weeks depending
on the microorganism ( Schmedes SE et al., J Clin Microbiol. 54(8), August

2016).
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AMPLICON SEQUENCING
The 16S rRNA gene is common among prokaryotes, and it is essential for bacterial life,
but it is not present in eukaryotes. This gene, which encodes the small subunit of the bacterial
ribosome, is characterized by species-specific variable regions, which are useful for identifying
phylogenetic relationships. After sequencing, 16S regions may be analyzed bioinformatically.
Highly similar sequences are grouped into operational taxonomic units (OTUs) with an allowed
typical degree of sequence divergence of 95%, 97%, or 99%. Since named species genomes are
often unavailable for particular marker sequences, OTUs take the place of “species” in many
microbiome diversity analyses. The community can be described in terms of which OTUs are
present, their relative abundance, and/or their phylogenetic relationships (Tozzo P et al.,
Microorganisms 8(6), June 2020).
METAGENOMICS
Metagenomics is the method of sequencing all microbial genomes within a sample. The
information obtained in this method can be used to identify which taxa are present and the
relative abundance of each, or to identify gene coding sequences and analyze the functional
potential of the microbial community (Tozzo, Pamela et al. Microorganisms vol. 8,6 ,Jun 2020).
In addition to targeted pathogen detection from samples for epidemiological and
biodefense purposes, metagenomic analyses of whole microbial community profiling hold
promise for microbial forensic utility.
Forensic metagenomics detects target microorganisms in complex samples. There are
currently two main approaches for metagenomic sequencing, targeting the 16S rRNA gene or
WGSS. The former provides better depth of coverage but tends to lack species-level resolution.
The latter can provide species or even sub-species level identification but lacks the depth of
coverage provided by targeted amplicon sequencing, which limits the sensitivity of detection of
target microbes. However, WGSS is more desirable for microbial forensic metagenomics
analysis as species level resolution is imperative (Schmedes S and Budowle B.,Encyclopedia of
Microbiology, August 2019).
BIOINFORMATICS
Bioinformatics is the application of computational methods to analyze and interpret MPS
sequence data, phylogenetic reconstruction, statistical analyses, and visual representation of data.
Numerous bioinformatics software tools and data management systems have been developed
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such as software tools for metagenomic assembly, taxonomic classification, phylogenetic

analysis, entire metagenomic analysis pipelines, database analysis and management systems.
These programs were developed for species- and strain-level detection of pathogens or other
target microorganisms from shotgun sequencing data with direct application for clinical
diagnostics and/or microbial forensics. Customized bioinformatics tools and comprehensive
databases for microbial forensic purposes are essential ( Schmedes SE et al., J Clin Microbiol.
54(8), August 2016).
MICROBIAL CLOCK
The PMI estimation is very important in reconstructing death scene and for issuing legal
certificates.“Estimating the time of death is basically an exercise in very broad probabilities,”
says forensic pathologist, Carl Schmidt of the University of Michigan [Carolyn Beans, PNAS
115 (1),January 2, 2018] . Each method, be it electronic or biological, has limitations and varying
accuracy depending on the environment and timeframe [Metcalf JL,Forensic Science
International: Genetics 38, January 2019].
Overview of the forensic tools available throughout timeline of human decomposition.
This microbial clock of death is developed by building a regression model using
microbiome data collected from postmortem samples (e.g.swab of skin, gastrointestinal, oral
/sample of internal organs, bone and soil) with known PMIs. In a death investigation, a similar
sample type (e.g. swab of skin) would be collected, the microbes profiled using DNA
sequencing, and the microbes would be matched to a point on the clock (i.e. the regression
model).
Machine learning is a powerful tool which is commonly applied to microbiome datasets
because they are large and complex. Amplicon sequencing data sets of taxonomically
informative, universal genes (16S rRNA, 18S rRNA, ITS gene region) provide a snapshot of the
microbial taxa in a sample, and are useful for creating regression models.
For example, they use the DNA sequences of a bacterial gene known as 16S rRNA to
identify and gauge the relative abundance of species in each sample and a gene known as 18S
rRNA for eukaryotic microbes such as nematodes and amoeba. In order to build a robust model
to estimate PMI, hundreds to thousands of samples with known PMIs are required. Additionally,
environmental data such as temperature, ecology and soil can be included in the model in a
similar way to the relative abundance of microbial taxa, and subsequently tested to assess
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whether they improve the model[Metcalf JL,Forensic Science International: Genetics 38, January
2019].The microbial successions were found to be remarkably consistent across different human
bodies and different seasons.
Carl schmidt and Jen Pechal studied that the bodies that have been dead for the same
period of time tend to have the same microbial presence and activity and cause of death for many
of the bodies such as homicide, suicide, accident, or natural death may shift the microbial clock
[Carolyn Beans, PNAS 115 (1),January 2, 2018].
The Human Microbiome Project (HMP) launched in 2014, provided enough evidence
that the microbial communities hosted by each individual is distinct and this fact is exploited in
forensics to strengthen evidences by linking the perpetrator with the crime scene/victim and
solve cases. A lot of research is still underway to uncover the enormous potential of microbial
forensicsand the technical advancements that are time efficient and affordable have made it
possible to a certain extent.
With the further development of microbial reference database, microbes can prove to be
an indigenous tool in forensics.
REFERENCES
1) Harbison S, Fleming R. “Forensic body fluid identification: state of the art”,Research and
Reports in Forensic Medical Science, February 2016;6:11-23 ;
https://doi.org/10.2147/RRFMS.S57994
2) Grice, Elizabeth A, and Julia A Segre. “The skin microbiome.” Nature reviews.
Microbiology vol. 9,4 (2011): 244-53. doi:10.1038/nrmicro2537
3) Schmedes SE, Sajantila A, Budowle B,“Expansion of Microbial Forensics”,J Clin
Microbiol. February 2016;54(8):1964-1974. doi:10.1128/JCM.00046-16
4) Oliveira, M., Amorim, A.,“Microbial forensics: new breakthroughs and future
prospects”,ApplMicrobiolBiotechnol 102, 10377–10391 (October 2018).
https://doi.org/10.1007/s00253-018-9414-6.
5) Tozzo P, D'Angiolella G, Brun P, Castagliuolo I, Gino S, Caenazzo L.,“Skin Microbiome
Analysis for Forensic Human Identification: What Do We Know So Far?”,Microorganisms.
2020;8(6):873. Published 2020 Jun 9. doi:10.3390/microorganisms8060873
6) Schmedes S, Budowle B.,“Microbial Forensics”,Encyclopedia of Microbiology.
2019;134-145. doi:10.1016/B978-0-12-801238-3.02483-1
ISBN: 978-81-948129-1-3 Page 496
7) Jessica L.Metcalf,“Estimating the postmortem interval using microbes: Knowledge gaps

and a path to technology adoption”,Forensic Science International: Genetics Volume 38, January
2019, Pages 211-218, https://doi.org/10.1016/j.fsigen.2018.11.004
8) Carolyn Beans, “News Feature: Can microbes keep time for forensic investigators?”
,PNAS ,January 2, 2018 115 (1) 3-6; https://doi.org/10.1073/pnas.1718156114
ISBN: 978-81-948129-1-3 Page 497

AN EMPIRICAL STUDY ON FAMILY PLANNING: THE HOUSEHOLD

CONSUMPTION AND SIZE
Dr. Pralay Ganguly
NSHM Knowledge Campus
Durgapur- W.B
ABSTRACT:
Family planning is a health measure, which is regarded as effective in the developing
world. Looking at so many works available, there is a lack of empirical work showing the
utilization of family planning on household composition/family size. As a result of that, this
study was conducted with the aim/objective of investigating the effect of family planning use on
the household composition in south 24 parganas, W.B. A quantitative means of data collection
was employed among a sample of 200 participants. A random sampling technique was used to
select the respondents. A pre-tested interviewer administered structured questionnaire and an
interview guide was used to collect data on the effect of family planning use on composition.
Data were analyzed using Mini tab and Excel. Findings from the research shows that using the
opinion of the respondents, there is a high level of neglect towards family planning as a result of
low level of formal education in Durgapur, W.B. This research recommends that there is need for
increase in the level of education among married couples so as to avoid the ignorance of family
planning method among them. It is also recommended that there should be an increase in the
income rate of households to encourage family planning.
KEYWORDS: Household, family size, composition, planning, development
INTRODUCTION
A household is one or more people usually resident in the same dwelling space living
facilities. A household can contain one or more families, or can contain no families at all. A
household that does not contain a family nucleus could contain unrelated people, related people,
or could simply be a person living alone. The household is one of the most important human
socioeconomic institutions, with functions that vary both across regions and over time. In many
historical populations, a household was not only a residence group but also a socioeconomic unit
within which production, consumption, reproduction, early childhood socialization, and many
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other activities took place. Many of these functions are still important in contemporary
households. It is this multifaceted character that makes the household an appealing subject of
study to many social scientists.
Household composition is a derived variable that classifies all households according to the
relationships between the people in them, and whether there is a family nucleus present or not.
India is the most populous country in Asia, with more than 135 crore people.
The annual population growth rate and fertility rate for women is quite higher side. This
has posed a major health and economic challenges to the nation. Family planning is immediately
relevant to development because it influences the birth rate and population growth rate of a
country, and therefore has far reaching impacts on economic, social, and environmental well-
being in society (Forbes. K., 2000; WHO, 2015a). Family planning goes a long way in helping
the problems being faced in households and family composition. Family planning has been
identified as a productive health right (Asgar et al, 2010). Family planning is a system which has
self-sufficiency and social development as its ultimate goals. It operates through the regulation of
fertility by the provision of birth control service.
There are so many important reasons why family planning should be taken seriously and
practiced by every society. It is a program aimed at the reduction of population growth rates to a
level compatible with the social economic aspiration of a society. In the writings of Tsui et al
(2010), research from various studies finds family planning interventions to be highly cost-
effective as they reduce the future health care costs associated with unintended pregnancies. The
challenges of households in carrying out family planning in south 24 parganas, W.B and how
this problem can be tackled is the reason behind this study “household composition/ family size
and the utilization of family planning in South 24 parganas”.
LITERATURE REVIEW
This article presents a review of research literature regarding the association between
family planning and household composition. So many authors have been looked up to in the
cause of understanding previous researches and better understanding of researches so far with
regards to the topic at hand.
Household and family composition is traditionally measured relative to the number of

households or families in the population. Thus, under the Census Bureau approach, one might
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measure the percentage of family households containing related subfamilies. Similarly, if one
employed the widely used Laslett-Hammel classification scheme, one might measure the
percentage of households containing multiple "conjugal family units" (Laslett in Steven and
Susan, 2003).
In a research carried out by Ranjan and Meenaksi 2000, there is a basic effort to
understand household size, family composition and its effect on poverty experienced by family
members. In summarizing the result of their findings, Ranjan and Meenaksi suggest that the
household size economies parameter cannot be relied upon to satisfactorily pick up household
composition effects. The head count poverty rates fall, quite sharply in many cases, with the
introduction of the State specific consumption economies of household size and of adult/child
relativities in the equivalence scale used as the expenditure deflator. AmsaleTekleErgano et al
(2016) carried out a research on the topic “the effect of family planning use on the household’s
economy”. At the end of the research, it was discovered that the proportion of women who
observed and reported the overall increase on household income attributed to the use of F/P was
only 41.8%. The type of family planning method, the total number of children, the frequency of
hospitalization and the reason for using family planning were some of the potential predictors of
household income. They recommended that a strategy has to be designed to reach women who
had less awareness about the effect of family planning in improving household economy. The
study found that women with access to family planning as teenagers gained 0.05 more years of
schooling, were 7% more likely to work in the formal sector, and were 2% less likely to cohabit
with male partners (Ellen Starbird et al 2016).
METHODOLOGY
AREA OF STUDY
South 24 Parganas (Pron: pɔrɡɔnɔs) (abv. 24 PGS (S)) or sometimes South Twenty Four
Parganas is a district in the Indian state of West Bengal, headquartered in Alipore. It is the
largest district of West Bengal by area and second largest by population. It is the sixth most
populous district in India (out of 640). On one side is the urban fringe of Kolkata and on the
other, the remote riverine villages in the Sunderbans.
The district comprises five subdivisions like- Aliporesadar, Baruipur, Diamond Harbour,
Canning and Kakdwip. Other than the municipality areas, each subdivision contains community
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development blocks which in turn are divided into rural areas and census towns. In total there are
118 urban units: 7 municipalities and 111 census towns.
According to the 2011 census of India, South 24 Parganas district had a total population
of 8,161,961,roughly equal to the nation of Honduras or the US state of Virginia. This made in
the 6th most populous district in India out of a total of 640.The district had a population density
of 819 inhabitants per square kilometre (2,120/sq mi). Its population growth rate over the decade
2001–2011 was 18.05%. South 24 Parganas had a sex ratio of 949 females for every 1000
males, and a literacy rate of 78.57%.
POPULATION OF STUDY
The sample size for this study is two hundred (200) and the two hundred (200)
participants will be administered a questionnaire. The reason for his choice of sample size is to
accommodate a quite segment of south 24 parganas local government area inhabitants and also to
give all the communities in this area equal chance of being represented. The sample technique
that was used in this study is simple random sampling. South 24 parganas consist of seven
communities, out of this communities, three communities were selected using simple random
sampling. Out of the three communities, eight villages were selected from five sub divisions (two
each from 3 sub divisions and one each from rest two subdivisions as per the population and
researchers judgement).
The households in the selected villages were numbered and out of it, 25 households were
selected from each village, making it 200 (196 finally administered after removing the sampling
error). The researcher made use of questionnaire schedule, which was fashioned out of the
research questions. The questionnaire has two parts. Part A: is made up of the personal data of
the respondent while Part B, deals with the substantive issues relating to the study.
DATA COLLECTION
The researcher made use of questionnaire schedule, which was fashioned out of the
research questions. The questionnaire has two parts. Part A: is made up of the personal data of
the respondent while Part B, deals with the substantive issues relating to the study. Selected and
trained research assistants (students of sociology department) administered the questionnaire
individually to the respondents on a face-to-face basis.
The respondents were assured of the anonymity of their responses as requested by them.
Some questionnaires was distributed, filled and collected at the spot while some respondents
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were allow some time to go through the questions at their free time with an agreed time of
collection. The data collected from the field were analysed using Mini tab and Excel.
DATA ANALYSIS AND INTERPRETATION
This chapter is a presentation, analysis and discussion of the finding on household
composition/ family size and utilization of family planning in Awka South. Tables, frequencies
and percentages have been used to present these findings. The discussion of these findings has
been organized specifically to answer the research questions.
Demographic characteristics of respondents. The demographic characteristics of respondents in
this section include genders, age, marital status, education qualification and occupation.
TABLE 1: SHOWING RESPONDENT GENDER
GENDER NUMBER PERCENT
Male 116 59.2%
Female 80 40.8 %
Total 196 100%
Source: Field Survey
From table 1 above, it was indicated that 59.2% of the respondents were male while
40.8% were female. This implies that there are more male than female respondents.
TABLE 2: SHOWING RESPONDENTS MARITAL STATUS

ITEMS Number Percent
Single 80 40.8%
Married 98 50%
Divorced 8 4.10%
Widow 105.1%
Total 196 100%
Source: Field Survey
From table 2 above it was indicated that the largest numbers of the respondents are
married with proportionate of 50% follow by 40.8% that are single while 4.10% of the
respondents are divorced and 5.1% are widow respectively.
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Table 3: Educated families adopt family planning method than non-educated families
RESPONSES FREQUENCY
PERCENTAGE
Yes 142
72.4%
No 34 17.3%
Don’t Know 20 10.3%
TOTAL 196
100%
SOURCE: Field survey
From table 3 above shows that 72.4% of the respondent accepted that educated families usually
adopt family planning method than non-educated while 17.3% did not accept this assertion but
10.3% could not say anything.
Table 4: problems facing household composition or family size.

RESPONSES FREQUENCY PERCENTAGE
Level of Income 48 24.9%
Religious believe 28 14.2%
Educational background 52 26.5 %
All of the Above 64 32.6%
Others 2 1.8%
TOTAL 196 100%
From table 4 above it was discovered that 24.9% admitted that level of income is one of the
problems facing household composition or family size while 14.2% suggested that it is as a
result of religious believe while 26.5% says it is educational background, 32.6% suggests all of
the above option and 1.8% suggested some other factors.
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Table 5: Possible solution to the challenges facing household in the management of family
planning
RESPONSES FREQUENCY PERCENTAGE

Increased income 22 11.2%
Increased Education Level 38 19.4%
Educative Programs ’ 40 20.6%
Government Support 28 14.2%
All of the above 66 33.6%
Other 2 1%
TOTAL 196 100%
From table 5 above it was discovered that 11.2% of our respondents says increased income is
one of the possible solution to the challenges facing household in the management of family
planning. Total, 19.4% says increased education level and 20.6% says educative programs’
could be possible solution and 14.2% suggest government support while 33.6% says all these
method factors could be used in addressing this problem.
CONCLUTION:
From the above analysis the researcher has come to the conclusion using the opinion of
the respondents, that there is a high level of neglect towards family planning as a result of low
level of formal education in South 24 parganas. It is also noticed that low income rate is one of
the biggest challenges facing households in the management of family planning. The various
ways by which this lingering issue can be solved or tackled is by increasing the educational level
of the people, organizing educative programs and governmental support.
RECOMMENDATION:
Base on the findings in the above analysis for the research work “household
composition/size and the utilization of family planning in South 24 parganas” the below
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recommendations have been out listed: 1. There is need for increase in the level of education
among married couples so as to avoid the ignorance of family planning method among them. 2.
There should be an increase in the income rate of households to encourage family planning 3.
Adoption or organization of educative programs and governmental/ institutional interference or
support will go a long way in controlling the challenges facing households in the management of
family planning.
REFERENCES-
1. Asgar, Md. Ali &Fazilah, A. & Huda, Nurul& Bhat, Rajeev & Karim, Alias. (2010).
Nonmeat Protein Alternatives as Meat Extenders and Meat Analogs. Comprehensive
Reviews in Food Science and Food Safety. 9. 513 - 529. 10.1111/j.1541-
4337.2010.00124.x.
2. Forbes, K. (2000)‘A reassessment of the relationship between inequality and growth’,
American Economic Review, 90(4): 869-887.
3. Tsui, Amy & McDonald-Mosley, Raegan & Burke, Anne. (2010). Tsui AO, McDonald-
Mosley R, Burke AE. Family planning and the burden of unintended pregnancies.
Epidemiologic reviews. 32. 152-74. 10.1093/epirev/mxq012.
4. Laslett in Steven and Susan, (2003). Management of household and family composition
in the United States. 1850-2000.
5. Meenakshi, J. V. & Ray, Ranjan, 2002. "Impact of household size and family
composition on poverty in rural India," Journal of Policy Modeling, Elsevier, vol. 24(6),
pages 539-559, October.
6. Ergano, Amsale. (2016). The Effect of Family Planning Use on Household Economy at
Arbaminch Town, GamoGoffa Zone, Southern Ethiopia. Science Journal of Public
Health. 4. 470. 10.11648/j.sjph.20160406.19.
7. Starbird, E., M. Norton, and R. Marcus. 2016. “Investing in Family Planning: Key to
Achieving the Sustainable Development Goals.” Global Health: Science and Practice
4(2): 191-210.
8. https://en.wikipedia.org/wiki/South_24_Parganas#cite_note-districtcensus-1
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LAWS AND ACTS FOR ENVIRONMENTAL PROTECTION

S.Rasul Meera and Dr. R. Kungumapriya
ABSTRACT
The present empirical research study focuses on the laws and acts for environmental
protection .To bring out the responsibility to protect our environment in a proper way by the
following last which has been constituted by the government. By using this various pollution has
been degraded, land grabbing can be eradicated, and wildlife animals could be protected. In the
society, awareness can be created such a way that ordinary people also can lead a healthy life.
There is a possibility to get clean air and clean water to lead our life in a healthy way. To create
awareness among the public, government has brought many acts by using it, we have to protect
the environment. To bring out the perfect hygiene in all over the place. There is a need for the
effective and efficient enforcement of law and environmental legislation. Poor sanitation
condition and sewage issues affecting the health of ordinary people in India. The reasons for this
disconnection between enlightened environmental laws and high levels of pollution could be
traced to last environment of already existing environmental laws, discrepancies in the
environmental guidelines for business to follow the between the Central and the State levels.
According to Article 21 constitution has provided Fundamental Right and Article 47 state that it
is primary duty of the state. To prevent noise pollution Motor Vehicle Act, 1988 has been
amended. Overall in this paper how the environment could be protected by using laws and acts
has been discussed.
KEYWORDS: Environmental protection, Sanitation, Article 21, Fundamental Right, Article 47,
1988.
INTRODUCTION
Environment includes all elements, factors, and conditions that have some impact on
growth and development of certain organism. Environment includes both biotic and abiotic
factors that have influence on observed organism. The word "environment" is most commonly
used describing "natural" environment and means the sum of all living and non-living things that
surround an organism, or group of organisms.
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Biotic factors such as light, temperature, water, atmospheric gases combine with biotic
factors (all surrounding living species). Environment often changes after some time and therefore
many organisms have ability to adapt to this alteration for their survival. However tolerance
range is not the same with all species and exposure to environmental conditions at the limit of an
certain organism's tolerance range represents environmental stress.
Environment is the immediate surrounding space around man. It includes a biotic and
biotic component. So environment not only means our environment but also varies of issues
connected with human activity and its impact on natural resources. It has been observed that in
recent past far reaching changes have taken place. Anthropogenic activities have played a key
role in altering the environment. Anthropogenic activities include over usage of own transports
instead abusing the public transport, Industrial wastewater and sewage water which water mixing
in Lakes and rivers to make them pollute harmfully.
Environmental law is a collective term encompassing aspects of the law that provide
protection to the environment. A related but distinct set of regulatory regimes, now strongly
influenced by environmental legal principles, focus on the management of specific natural
resources, such as forests, minerals, or fisheries.
ENVIRONMENTAL LAWS IN INDIA

In order to protect environment the constitution of India has provided a foundation for
framing environmental legislation. Environmental protection has become a major concern and
global issue with an increase in many environmental issues. To bring out the awareness about
our environment to the common people. Our five most effective pieces of environmental
legislation are the Clean Air Act, the Endangered Species Act, the Montreal Protocol, the Clean
Water Act, and Reformation Plan No. 3 of 1970. Because of these laws, the health of Americans
and the environment they inhabit have dramatically important. (Jul 1, 2010)
The number of polluted stretches in India's rivers has increased to 351 from 302 (in 2006), and
the number of critically polluted stretches where water quality indicators are the poorest - has
gone up to 45 from 35 (Source: The Hindu, 17 September, 2018).
CONSTITUTIONAL PROVISIONS RELATED TO ENVIRONMENT
Following are the constitutional provisions that deal with the environmental protection
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Article 21 of constitution has provided fundamental right viz., Right to life.

 Supreme Court of India has included the right to healthy environment within the
Ambit of right to life. These types can be interpreted and used as tool for
sustainable development.
Article 47 states that it is primary duty of the state.
 to raise the standard living of the people
 to increase the nutritional level of the people
 to bring the improvement in public health.
Article 48 (A) of constitution state that the state has to protect and improve the
environment and to safeguard the forest and wildlife and improve the natural
environment.
Article 51 (A) states that citizens are required to protect and improve the natural
environment, including forest Lake Rivers and wildlife. With the constitution various
environmental legislation have been framed by state and union governments and
pollution control board to regulate environmental pollution.
ACTS RELATED TO ENVIRONMENT PROTECTION INDIAN FOREST ACT, 1927
This Act consolidates the law relating to forest, the transit of forest produce and the duty
liveable on timber and other forest produce. Provision related to our forests under the Act are as
follows • It provides power to State Government to constitute any forest land, waste land
(property of government) as reserved forest State can appoint a Forest Settlement Officer to
a. Inquire over a reserved area Provisions related to Village Forest under the Act are as
follows,
 The State Government may assign to any village community the rights of government
to of over any land which has been constituted as reserved. All forests so assigned
shall be called village forests.
 The community to which such assignment is made may be provided with timber or
other forest produce or pasture and their duties for the protection and improvement of
such forest.
b. Provisions related to Protected Forests under the act are as follow
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 Prohibition of quarries stories, booming lime, charcoal or removal of any forest

produce, clearing land for cultivation building or any other purpose in any protected
forest.
 Central Government can impose duty on timber and other forest produce.
 Cattle trespassing in a reserved forest or any portion of protected forest may be seized
 Any person violating any rule under this Act shall be punishable with imprisonment
for six months or fine extending up to 500 or both.
WILDLIFE PROTECTION ACT, 1972
Wildlife plays very important role in conservation and maintenance of ecological
balance. The first comprehensive legislation related to the conservation and protection of wildlife
was passed by Government of India are 9th September 1972. This act was amended in year of
1982, 1986, 1991 and 1993 respectively. It deals with the regulation of a hunting and trade in
wildlife.
This Act was framed to protect and conserve wildlife which includes animals, birds and
plants. This act is implemented throughout India except the State of Jammu and Kashmir.
OBJECTIVES OF WILDLIFE PROTECTION ACT, 1972.
Biodiversity preservation and protection - Protection of wildlife sanctuaries. National
park an drivers, reserve.
Maintenance of infrastructure and strengthening management of national parks and
sanctuaries.
FORMATION OF WILDLIFE ADVISORY BOARD

Wildlife Advisory Board is constituted under Wildlife Protection Act, 1972. Wildlife
Advisory Board is constituted under guidance of State Government. It comes to following means
 Minister In charge of Forest in the State or Union territory or Secretary of the State
Government shall be the Chairman
 2 members of State Legislature and UT having legislate
 Secretary to State Government In charge of Forests.
 Forest Officer In charge of the State Forest Department.
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 1 officer nominated by the Director

 Not more than 5 officer of State Government
 Chief Wildlife Warden.
CONCLUSIONS AND RECOMMENDATIONS

Environment is an important part of human life and a healthy environment is a must for
human existence. Therefore, it is crucial that we take care of our surroundings and help nature
maintain ecological balance so that we could hand over to the coming generations the
environment as we found it, if not any better. In the recent past there has been a lot of damage to
the ecology. Air, water and soil have been polluted and there appears to be no decisive end to it.
The scientific advancement and rapid industrialization has taken its toll.
Nowadays protection of the environment is very important as the world is moving into a
new era without considering any of the major problems of pollution with rapid industrialization.
The best way to protect the environment is conservation. Conservation is the philosophy and
policy of managing the environment to assure adequate supplies of natural resources for future as
present.
Tropical forests are being destroyed at an ever-increasing rate. Estimates of the extent
and rate of loss vary, but it appears that nearly half of the world tropical forests already have
been lost, and the remainder will all but disappear in the next two to three decades. The loss is
incalculable.
These forests provide habitat for an estimated half of the world plant and animal species,
provide water and fuel for much of the world population, and influence regional and global
climate.
COMMERCIAL LOGGING, CLEARANCE FOR AGRICULTURE
The Courts in India have played a distinguishing role in gradually enlarging the scope of
a qualitative living by applying various issues of environmental protection. Consequently,
activities posing a major threat to the environment were curtailed so as to protect the individual’s
inherent right to wholesome environment. Article 21 has been relied in the plethora of cases,
although certain cases have incorporated a wider perspective of the Constitution. Hence, the
Supreme Court of India, apart from being environment friendly, has given birth to a wide range
of doctrines and principles have in turn been adopted and implemented throughout the country.
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The suggestions made by the Supreme Court in A.P. Pollution Control Board. M.V.Nayudu, for
the improvement of the adjudicatory machinery under the various environmental laws should be
implemented by the Government. The main burden of these suggestions is that in all
environmental courts, tribunals and appellate authorities, there should be a judge of the rank of a
High Court or a Supreme Court, sitting or retired, and a scientist or a group of scientists of high
ranking and experience so as to help a proper and fair adjudication of disputes relating to
environment and protection. If implemented, this would go a long way in securing justice to the
needy.
Any law is as good as the implementation. The implementation mechanism must be
foolproof and effective. There must be an effective monitoring mechanism. The bottom line is
that we must have individuals of integrity with a strong value base and deep commitment if laws
are to be effectively implemented. Therefore, protecting our environment is each and every
citizen's duty and responsibility. Have to appoint some special officers to prevent River sands
and Timber smuggling it’s one of the mandatory work done by the government. Finally we will
protect the environment and environment will protect us.
REFERENCES
1. LL.M Second year (Business Law Group), Rajiv Gandhi National University of Law,
Patiala (Punjab) Mohindra .
2. .Kothi, Mall road, Near Fountain chowk, E-mail:-
pradiprgnul@gmail.com,sunit1215@gmail.com.
3. L.D Saini, Environmental Education, Kalyani publisher, Ludhiana, p
4. NTA UGC Environmental Science Paper-2 Q019) : (S5552).
5. Text book of Biology - Zoology (2014), 219.
6. Pradip Kumar Kashyap&SuneetDwivedi, Environmental Protection Law and Policy in
India,
7. Elliott, J. (March 25, 2002). “Bhopal Refuses to Flip the Page: After More Than 17
Years, Thousands of Indians Still Suffer from the Lethal Gases That Leaked from a US
Chemical Plant”. New Statesman, 131. Retrieved December 23, 2011, from
Questia.com,1986.
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A COMPREHENSIVE REVIEW ON THE NANO- CONFINED AND ENGINEERED

NANOWOOD MATERIALS FOR WATER PURIFICATION PROCESS
V. Ragul1 and M.I. Niyas Ahamed*
Department of Biochemistry,
Sacred Heart College (Autonomous), Tirupattur, Tamil Nadu, India.
*driniyasahamed@shctpt.edu, 1ragulshc05@gmail.com
ABSTRACT
The wood materials are dynamically designed with extraordinary characteristics for
numerous application aspects. The simplified cutting edged nanobiomaterials transform with the
cheap, cost effective and easily available woods are considered as updated and emerged material
which is having its determined life time, structural property, and effective functions to the
alternative resource for different kinds of polymers, plastics and metals. The present model
expresses how the innovative nano wood materials have been developed and implemented in
contaminated water in different parameters with effective functionality were described and
discussed. The proper arrangement with hierarchical structures were introduced for
bioremediation of contaminated water by modified or unmodified either top down or bottom up
production of water filters with energy efficient desalination, catalytic activity, energy
investment, resource recovery and environmental cleanups were discussed. The investigation
offers, nascent but highly promising field to encourage research and development. Once the tree
is harvested and used, it is ultimately understood that enhanced wood materials will provide a
great impact in the circular water economy.
KEYWORDS: Bioremediation; Desalination; Wood materials; nano wood.
INTRODUCTION
All the existing creatures start from adequate and effective water, which act as a dynamic
element in the biological system. The consideration offers to the water security for worldwide is
wellbeing and monetary enhancement for guaranteeing pledge against water-borne
[1]
contamination and water-related fiascos would never be excessively . These days, the
expansion of the limit of modern creation, the interest of human movement and the water
contamination, we are confronting more genuine clean water shortage than any time in recent
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decades. Thus, an assortment of water treatment advances has been created to filter contaminated
water so as to accomplish reasonable water use [2,3].
Since, past 20th century, the flourishing explores on different bio-nanomaterials have
been changed our experience with numerous major procedures what's more, carried progressive
advancements to different building viewpoints. For models, the exposure of carbon based nano
biomaterials like, graphenes and carbon nanotubes has caught the creative mind of utilizing its
shocking optical and electronic properties for different application prospects [4]. The advancement
of nanolithography innovation permits the command over the shape, size and piece of structure
on the length of 1-100 nm with effective openings in various areas to running different clinical
diagnostics. This accomplishment in nanotechnology additionally improves new open doors for
water purification strategies. Recent reviews summarized representative examples
of how nanotechnology creates novel materials that could enrich some water treatment
frameworks with excellent properties, for example adsorptive, synergist, electrical or potentially
antimicrobial, that improve cost-efficiency[5-7].
A promising advance forward into the scaled-up utilization of these nanomaterials
requires their immobilization inside different substrates to shape composite materials, which are
alluded to as nanocomposites in this report. The substrate in the nanocomposites gives not just
minuscule space to the convenience of nanoparticles, yet additionally naturally visible shapeable
structure and processability. The nanosized spatial limitation gave by the substrate has been
named as nanoconfinement. More critically, nanoconfined water shows unusual properties
furthermore, practices, for example temperature height of the freezing transition and bizarrely
low dielectric constant[8]. Meanwhile, the vehicle of water and particles inside nanoconfined
channels doesn't follow the continuum transport model for bulk. As of late, propelled by the new
information picked up from the essential investigations on nanoconfinement, researchers have
attempted to investigate the use of nanoconfinement in different fields. For heterogeneous
catalysis, nanoconfinement with one of kind geometric and electronic structures has been utilized
to adequately balance the reactant execution of the restricted dynamic species [9,10]. Despite few
existing models indicating the improved toxin expulsion effectiveness under nanoconfinement, a
methodical comprehension on the connection between the extraordinary properties of materials
and water atoms under nanoconfinement and the poison expulsion is in pressing need. In this
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audit, we might want to utilize a new point of view on how nanoconfinement could alter our
understandings on different procedures engaged with water treatment [11-13].
NANO-MANUFACTURING OFFERS VERSATILE STRUCTURAL DESIGNS AND

EFFECTIVE FUNCTIONS OF WOOD MATERIALS FOR WATER-DEPENDENT
APPLICATIONS
Wood possesses dynamic hierarchical constructions with a number of orders of
magnitudes to support several functions which plants promote for effective growth. The cell wall
of the plants are initially prepared by the composition of lignin, cellulose and hemicelluloses
construct micro fibrils with nanometer scale further the micro fibrils were arranged in the parallel
manner to provide structural support to the plant growth and development [14]. The cellulose
micro fibrils are basically arranged with the size of 3-10 nm width and 10-30 micrometer long. It
can be separately examined and isolated to become nanocellulose crystals/fibers. There
nanocellulose would be considered as building blocks with effective functional filaments,
membranes, films, aerogels and hydrogels. This existing nanocellulose are possessed and used in
the water treatment process. In addition it is used in the form of water filter membrane to
increase permeability, increased pressure, adsorption capability and improved elimination of
contaminants through interaction of nanoparticles with different modification processes.
Compared to the bottom-up nanocellulose emerged with a new top down approach. Vertical
dimension channels with the size of around 20-130 micrometers tree trunks were called vessels
which took up charge of water and tree crown[15-18].
Several microns were existing on the vessels of the walls which helped to disperse the
contaminated water and several ions in the water in radial direction in the trunk. The arranged
abundance of pits not only has anisotropic mechanical properties and also the water can transport
in multi directions. The naturally existing hierarchical mesoporous wood materials and its
structures provide improved separation of materials. Naturally a tree absorbs carbon dioxide and
harnesses the energy from the sun to carry out special structural properties [19,20]. Figure 1
demonstrates that functionalized wood based materials offer different water treatment
applications. In addition the alternated naturally existing wood materials shows a variety of
functions can be offered with prescribed catalytic, adsorptions, desalinations, filtrations and
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formation of energy. These functionalized nanomaterials which play an important role in

bioremediation of water to protect the economy of water[21].
Figure 1. Nanowood developmental, versatile parameters and dynamic function offers

various water dependent applications [22].
Table 1 - Different characteristic features of natural and engineered nanowood materials[22]

Natural wood Engineered nanowood
Characteristics Modification Characteristics Application
Nonconductive Carbonization Electro-conductive Energy generation,
storage, and delivery
Non-catalytic Particle Highly reactive Contaminant
immobilization degradation
Low light adsorbing Carbonization Enhanced light adsorbing Solar heating and
efficiency efficiency desalination
Hydrophilic Chemical treatment Hydrophobic Oil adsorption and
vapor transport
Mesoporous and small Delignification Increased porosity and Water and vapor
pores pore size transport
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Figure 2: Different characteristic parameters of naturally existing mesoporous wood

materials[22].
CELLULOSE MEDIATED MEMBRANE FILTER FOR WATER TREATMENT
The chief characteristic of wood is cellulose nanomaterials including cellulose and

nanocellulose dependent nanofibers are effectively used as membrane filtration fabrication
processes. When compared to all other nanomaterials cellulose mediated nanomaterial and
carbon nanotubes are considered eco friendly, cost effective and renewable material. In the
cellulose dependent nanomaterials carry out unique features with increased hydroxyl groups and
effective mechanical strength[24]. Cellulose acetate and cellulose derivatives were first mixed in
organic solvents and made as desalination membranes in the 1950s and offers reverse osmosis
for commercial processes. Nanosized materials offer great attention to improve the permeability
and increased hydrophilicity and smooth surface to decreased membrane fouling. It has strong
mechanical property and it can withstand high pressure. Number of studies expressed that
cellulose mediated materials offer great attention towards membrane filter processes which
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were considered emerging innovative strategies in the water economy. It includes nanofiltration,
ultra filtration and forward osmosis were expressed in table 2. Cellulose mediated membrane
filters can be formed by using different methods to get desired functional goals. It may include
very common blending , vacuum filter, phase inversion and also interfacial polymerization.
The eminent scientist fabricated based on cellulose based membrane from wood which offers
great attention on bioremediation of water with different parameters [25-29].
Table 2: Summary on cellulose mediated membrane for water treatment [30]

Materials Process Feed Fabrication Rejectio Other Permeabilit
Solution Process n y
Vacuum
filtration and
Cellulose Ultra 1000 ppm 97% Less than 5%
interfacial
nanocrystal filtration Na2SO4 rejection of
polymerizatio 34 LMH/bar
NaCl
n
80% of
Spin coating sucrose
Hydrophobic Ultra 1000 ppm Nearly 0%
with
trimethylsilyl filtration sucrose rejection of
MWCO 1.5 LMH/bar
cellulose NaCl
of 342
MWCO
Poly(ethyl Spin coating between
Nanofibrillated Ultra ene Pure cellulose 4
(6 & 26
cellulose filtration glycol) material LMH/MPa
kDa)
Bovine 40% of Fouling

serum BSA resistance
Hydrophilic Ultra Blending-phase
albumin (MWCO decreased by
filtration inversion
cellulose (BSA) - 68000) 48.8% 21
nanocrystal solution LMH/MPa
Ag- and Pt- 5% w/v 1.2–1.4 Electrochemic

decorated NaCl Nonsolvent gMH al and
Flow induced phase 10.24
nanocellulose draw (reverse antimicrobial
operatio separation LMH/bar
solution salt flux) reactive
n
High tensile
DI water Blending and strength of
Cellulose Ultra phase inversion - 7048
2.6 MPa
nanocrystal filtration LMH/MPa
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The movement of water in the membrane was used as irreversible fouling resistance
which was decreased by 48.8 % that treated with modified modelfoulant BSA were represented
in the figure 3 a and b. The elimination or removal of nano membranes towards BSA decreased
in the ranges around 90 to 40 percent. The different characteristic study was determined and
expressed in the figure 3 a - f. The improved physical and mechanical strength of the obtained
membrane were stable removal of monovalent ions like NaCl. The activity of the membrane
was observed in increased level when compared with commercially available membranes. In
addition the elimination of NaCl was achieved by using commercially existing membranes like
NF and it shows great attention and approximately zero NaCl elimination with the molecular
weight of 300 Da. In the membrane filtration process nanocellulose offers enhanced
performance of FO membrane filters[31,32]. The increased aspects of nanocellulose biomaterials
allowed improved density entrapped and FO membrane synthesized with Pt and Ag
nanoparticles entrapped nanocellulose as additional sources. The NF and FO membrane shows
great antimicrobial activity and electrochemically reactive. Desalination of these fabricated
membranes poses around 60% of increased water influx and out flux in commercially available
membranes[33].
Figure 3. a) Movement of water in cellulose membrane, b,c) BSA filtration at different

CNC in before and after membrane fluxes and rejections, d) CNC and CNT entrapped
ultrafiltration membrane, e) NF and CNC interacted membrane, f) Na 2SO4 and NaCl removal
at different salt concentration[34-37]
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CATALYTIC CONTAMINANT ELIMINATION OF MESOPOROUS BULK WOOD

STRUCTURES
The properties of the cellulose mediated membrane filter shows great intensive
application on waste water treatment. The narrow utilization of modified wood based
materials offers great attention in number of fields. The meso-structured natural and
fabricated wood with extended channels shows huge impact in the bioremediation process of
water and translocation of different ions by the interaction between the wood surface area
and water. The increased surface area of the wood posses huge number of hydroxyl groups in
the cellulose acts as active sites for catalyst immobilization and catalyst [38]. When the wood
is allowed to interact with targeted nanoparticles shows great impact in the bioremediation
process of contaminated water was performed in situ degradation of contaminants in the
water were expressed in table 3. The summery expressed how the different nanomaterial
mediated and fabricated wood materials shows improved micro reactors for the elimination
of wastes in the water. The existing challenges and advantages offers either oxidation or
reduction process[39].
Table 3: Different characteristic property of catalytic or adsorptive nanowood reactors and

materials for reduction of pollutants[22]
Process Contaminent Modified Removal Performance

Material mechanism
Batch reactor 30 ppm Carbonized Chemical Up to 99.8%

methylene blue basswood + Fe– reduction with removal
(MB) Mn–O nanosheet NaBH4
Flow-through 10 ppm Chemical Up to 98.5% MB

reactor Methylene blue reduction with elimination 5.2–6
E. coli and Radiate pine NaBH4 orders pathogen
S. aureus sapwood elimination
(softwood) + Ag
NPs
Chemical
Flow-through Carbonized first
10 ppm MB oxidation with Up to 95% removal
wood + TiO2 and
reactor H 2O 2
Mn O NPs
Flow-through 30 ppm Basswood + Pd Chemical Up to 99.8%

reactor methylene blue NPs reduction with elimination
(MB) NaBH4
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Initially the emergence of nanoparticles (Pd NPs) wood based water purification was
achieved by performing methylene blue as a contaminated sample. The obtained wood pieces
were socked and heated into the Pd NPs solution for 12 hours at 80⁰ C. for unique deposition or
interaction of nanoparticles to the wood sample. From this the Pd ion reduction was achieved by
lignin and more stable immobilization by hydroxyl groups. Then, the prepared methylene blue
sample was passed into the fabricated wood material. By this action Pd activates H 2 which acts
as an effective reducing agent for contaminant reduction. NaBH4 was allowed to interact with
water and effectively collected H2 [40].
From the obtained sample it was confirmed that the bioremediation was achieved around
99.8 % with the initial concentration of 30 ppm was eliminated and a maximum or increased
flux ratio of 105 LMH which was represented in the figure 4 c&d. The conversion of reactant by
the wood material was achieved around 2 molar of methylene blue mol Pd -1 min-1. In addition
silver nanoparticles were incorporated with wood membrane shows great attention toward
bioremediation of contaminated water and also it shows improved activity towards filter the
microorganisms like E.coli and S.aureus which was represented in figure 4 e -g. from these
investigation revealed that, the bioremediation of contaminated water is still in emerging
condition and it requires more attention to the investigators to eliminate the contaminants in the
drinking water[41-44].
ISBN: 978-81-948129-1-3 Page 520

Figure 4: Elimination of contaminants by nanowood materials a) mesoporous figure of

naturally existing wood, b) Bioremediation of MB by Pd NPs incorporated wood material, c)
property of MB elimination, d) permeate fluxes, e) OD for different bacterial consortium with
different time, f,g) Mn3O4/ TiO2 decorated wood under several magnification (SEM), h-j) Mn,
Ti and O mapping[45] .
CHEMICAL ADSORPTION FOR FUNCTIONALIZED AND CELLULOSIC

COMPOSITE IN BULK WOOD MATERIALS
Total wood segments and cellulosic composites show improved mechanical strength and
good porous structure and increased surface area. These properties shows unique for chemical
and physical adsorption of different pollutants like dyes, hydrocarbons, heavy metals and other
mixed organic compounds. The existing of hydroxyl group in the wood material offers
increased adsorption capability to the desired contaminants. From the figure 5 the cellulosic
fiber mediated nanofiber adsorption studies on heavy metals in after and before interaction [46].
Figure 5: Characteristic image of cellulosic nanofiber mediated filter for adsorption of heavy
metals by before and after interaction[47].
The emerging investigations make a key note to the bioremediation of heavy metals, oil
traces and other toxic contaminants. Those obtained and prepared natural or fabricated wood
materials shows improved tensile strength of 5.19MPa and increased binding ability to Cd like
heavy metals around 25-10 mg g-1. Modification of hydroxyl group in the cellulosic membrane
modifies the adsorption capacity of the heavy metals. As determined above effective wood
materials shows preliminarily investigated for oil, heavy metals and other toxic contaminants
adsorption studies. In addition simple modified chemicals have been investigated and the
ISBN: 978-81-948129-1-3 Page 521

obtained materials may deal different difficulties in the multiple pollutants in tested samples [48].
The efficiency was noted and it was optimized with standard samples in the fabricated and
natural samples. The increasing concentration of co-ions like sodium and potassium in water
may decrease the binding efficiency of heavy metals in the wood membrane [49]. The different
electro active and electro conductive bacterial attractions were shown in the outer surface of the
wood materials were observed in the figure 6.
Figure 6: Different electo-conductive and electro active bacterial adsorption studies [50]
CONCLUSION
With expanding uneasiness encompassing the exhaustion of common assets and
crumbling of the earth, more consideration has been coordinated to manageable turn of events, so
as to safeguard our planet for people in the future. With its huge and inexhaustible crude material
flexibly, the woodland can furnish us with materials that can be utilized in about each part of our
day by day life to supplant impractical and vitality concentrated man-made materials. Changing
to ecologically amicable, inexhaustible, and reasonable wood-based crude materials has pulled in
enormous intrigue, and critical advancement has been constructed as wood depended progressive
confinements. In view of beginning nanomaterials, two dynamic inverse plan methodologies has
been evaluated and summed up here.
From this review, it gives an outline of different minimal effort adsorbents instead of
costly business adsorbents utilized for the viable expulsion of fluoride from water. The
effectiveness of various adsorbents is relying upon boundaries, for example, pH, adsorbent
portion, surface territory, contact time, temperature and introductory fluoride fixation. The
ISBN: 978-81-948129-1-3 Page 522

expulsion limit increments by expanding portion of the adsorbent and diminishing size of the
adsorbent. The WHO norms endorse 1.5 ppm as most extreme level of fluoride in drinking
water. The future examination ought to be focused on improvement of powerful and financial
defluoridation adsorbents with legitimate local or network units for creating nations as far as cost
and practicality for the evacuation of fluoride.
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51. M. Barathi, A. S. K. Kumar, and N. Rajesh,. 2019. “Impact of fluoride in potable water -
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POLYMORPHISMS IN DNA REPAIR GENES IN CARCINOMA PATIENTS LIVING

IN A VERY COAL-MINING REGION
Shubham
Air pollutants and radiation are well-known carcinogens involved within the pathogenesis
of carcinoma, and residents of coal-mining regions are exposed routinely to those agents.
Polymorphisms in DNA repair genes is also related to an increased risk of malignant
transformation. We investigated associations between the chance of carcinoma in residents of the
coal-mining region and polymorphisms within the genes APEX1 (rs1130409), hOGG1
(rs1052133), XRCC1 (rs25489, rs25487), XRCC2 (rs3218536), XRCC3 (rs861539),
ADPRT/PARP1 (rs1136410), XPD/ERCC2 (rs13181), XPG/ERCC5 (rs17655), XPC
(rs2228001), АТМ (rs1801516), and NBS1 (rs1805794). 300 and forty residents of the
Kemerovo Region (a coal-mining region of western Siberia) were carcinoma patients exposed to
air pollutants and radiation (case) and 335 were healthy donors (control). Genotyping was
performed by real-time PCR and allele-specific PCR.
We discovered that polymorphisms within the XPD gene in men [log-additive model:
odds ratio (OR) = 1.64, 95% confidence interval (CI): 1.17-2.31], the ATM gene in women and
nonsmokers (codominant model: OR = 0.11, 95% CI: 0.02-0.49 and OR = 0.25, 95% CI: 0.08-
0.72, respectively), the APEX1 gene for smokers (recessive model: OR = 2.55, 95% CI: 1.34-
4.85), and also the NBS1 gene for people who add the industry (overdominant model: OR =
0.40, 95% CI: 0.21-0.75) are related to an increased risk of carcinoma. Using the multifactor
dimensionality reduction method, we found a model of gene-gene interactions related to the
chance of lung cancer: NBS1 (rs1805794)-XRCC1 (rs25487)-hOGG1 (rs1052133)-XPG
(rs17655). These results indicate an association between combinations of polymorphisms within
the studied genes and also the risk of carcinoma in residents of a coal-mining region.
ISBN: 978-81-948129-1-3 Page 529

MEDICINAL TREE - Couroupita guianensis

Vino Udappusamy
Research scholar, Department of Biochemistry, PSG College of Arts & Science,(Autonomous),
Coimbatore.
E.mail:u.vino95@gmail.com
Plants are the basic source of modern medicine. Almost all the parts of plant, namely
leaves, flowers, fruits, roots, stem and seeds are known to have various medicinal properties. The
trend of using natural products has increased and the active plant extracts are frequently screened
for new drug discoveries. One important plant that is used in traditional medicine is Couroupita
guianensis is a tree belonging to the family Lecythidaceae. It is native to South India and
Malaysia. In India, it is considered as a sacred tree by Hindus which is the reason why it is
generally grown in Lord Shiva temple because of its special featured flowers which look like
hood of Naga (snake) protecting the Shivalinga, hence, it is called as “Nagalinga Pushpa”.
Traditionally, ‘Kailashpati’ is widely used for its medicinal properties and also extensively
research for its pharmacological properties.
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ARTIFICIAL INTELLIGENCE IN AGRICULTURE: A MINI REVIEW

Atishya Mahesh Jain and Murugan S
Department of Biotechnology, School of Agriculture and Biosciences
Karunya Institute of Technology and Sciences (Deemed to be University)
Coimbatore- 641 114
*Email: murugan@karunya.edu
INTRODUCTION
The FAO (Food and Agriculture Organization) of the United Nations reports that in 2050
the world's population will rise by another 2 billion, although at that time the additional land area
under cultivation will account for just 4 per cent. Agriculture is significant, from generating jobs
to contributing to national income. It contributes significantly to the economic prosperity of
developed countries and also plays an active role in developing a country. Increased agriculture
has resulted in a substantial rise in the rural community's per capita income. Therefore, it would
be rational and appropriate to place greater emphasis on the agricultural sector. The biggest
concern and emerging issue around the world is automation in agriculture. The population is
rising tremendously and the demand for food and jobs is also growing with this increase. The
conventional methods used by the farmers were not adequate to satisfy these requirements. In
such situations, using recent technical developments and solutions to existing farming
bottlenecks, more productive farming practices can be accomplished.
A direct implementation in the agricultural sector of AI (Artificial Intelligence) or
machine intelligence could serve as an epitome of change in the way farming is practiced today.
AI-powered farming solutions allow a farmer to do more with less, improve quality, and also
ensure a fast GTM (go-to-market strategy) strategy for crops. These technologies save the
unnecessary use of water, pesticides and herbicides, preserve soil fertility, lead to the productive
use of human resources, increase productivity and improve efficiency. Equipment and machines
based on AI have brought today 's farming system to a new level. This technology has increased
crop production and increased tracking, harvesting, processing and marketing in real time. In an
attempt to make farming more precise and controllable, Precision Agriculture uses technical
advances such as Global Positioning System guidance, robots, control systems, drones,
autonomous vehicles, sensors, etc. The Global Precision Farming market accounted for $2.81
ISBN: 978-81-948129-1-3 Page 531
billion in 2014, according to Statistics MRC, and is projected to rise at a 12.5 percent CAGR to
reach $6.43 billion by 2022. In the agro-based industry, the new technology in automated
systems using agricultural robots and drones has made a significant contribution. Different hi-
tech computer-based systems are designed to recognise various important parameters such as
weed detection, crop quality and yield detection and many other techniques.
IMPACT OF AI ON AGRICULTURE
With the advent of technology in this digital environment, we humans have pushed our
limit of the thought process and are trying to coalesce with an artificial normal brain. The idea of
cognitive computing is one that, as a computer model, imitates the process of human thinking.
These results in chaotic technology in AI-powered agriculture, making its service to improve
efficiency in understanding, acquiring and responding to various situations (based on the
learning acquired). Deep learning, Convolutional Neural Network, Artificial Neural Network,
Machine learning are some domains that improve the work of the machine and help to develop
more advanced technology. In order to distinguish weeds from crops, Gliever and Slaughter used
ANN. For forecasting variables of water supply, Maier and Dandy used neural networks.AI-
based technologies help to increase productivity in all sectors and also manage the challenges
faced by different industries, including different fields in the agricultural sector, such as crop
yield, irrigation, soil content sensing, crop tracking, weeding, crop establishment. For daily
agricultural tasks such as harvesting, planting, seeding and so on, companies build self-
controlling robots that complete their tasks at a faster pace than human workers. For example, a
robot called "See & Spray" was created by the company "Blue River Technologies" to track and
spray weeds on cotton plants. Undoubtedly, one of the most important agricultural operations is
the management and maintenance of the soil and crop health. The Berlin-based start-up "PEAT"
has created an application called "Plantix" to replace humans and thus eliminate the possibility of
failures, which can analyze the health status of the soil.
The relationships between the different embedded systems and AI technology consistent
with the agricultural area, gave a brief on the different neural network applications and ML in
this sector for precision farming (Jha et al.,2019). According to Panpatte (2018), artificial
intelligence allows farmers to collect large quantities of government and public website
information, analyse all of it, and provide farmers with solutions to many ambiguous problems,
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as well as providing us with a smarter way of irrigation that results in higher yields. An FK-
based fuzzy model was developed by Sicat et al .,(2005) to determine the suitability of property.
An innovative method for grading leaf diseases was developed by Sannakki et al . ( 2011). In
their work, Ferguson et al . (1991) reported on the optimum production level for all plants was
set by variety selection and seed quality. Emerging technologies have led to the best variety of
crops and have also increased the option of hybrid seed choices that are best suited to the needs
of farmers.
In "Scaling up artificial intelligence agricultural research," Bestelmeyer et al.(2020)
developed AI-based tools that exploit site-based science and big data to help farmers and land
managers make site-specific choices. These instruments provide early warning of outbreaks of
pests and diseases and promote the selection of sustainable practices for cropland management.
In "Remote Sensing: Advancing Science and Applications to Transform Agriculture," Hatfield et
al.(2020) developed tools to identify variable areas within fields using remote sensing coupled
with neural networks and Machine learning and identify possible adaptive strategies to improve
profitability for each field while minimizing the environmental impact through more efficient
nutrient and pesticide usage. The researchers have now begun to emphasize technologies to
design autonomous agricultural tools as efficiency is lacking in traditional farming machinery.
The robots autonomously carry out various agricultural operations such as weeding, irrigation,
guarding the farms for efficient reporting, ensuring that adverse environmental conditions do not
affect production, incrustation.
In predicting the level of nutrition in the crop, Song and He (2005) brought together
expert systems and artificial neural networks. Using significantly less meteorological data, Arif
et al.(2012) built two ANN models to estimate soil moisture in Paddy fields. By observing the
projected soil moisture levels, all these models were then corroborated and validated. Drones for
crop health monitoring, irrigation equipment monitoring, weed identification, herd and wildlife
monitoring, and disaster management are being deployed in agriculture. The water to be fed to
the farms should be of good quality, either by means of smart irrigation or by some other
traditional method. In the aquaculture market, along with agriculture, researchers have begun
implementing IOT systems and artificial intelligence techniques. By deploying state-of-the-art
automation techniques, the system designed by Encinas et al.(2017) monitors water quality. Hunt
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et al. (2005) evaluated Digital Imaging for Remote Sensing of Crop Biomass and Nitrogen Status
from Model Aircraft.
CHALLENGES AND FUTURE SCOPE
Agriculture has been addressing major problems such as lack of irrigation systems,
climate rise, groundwater density, food shortage and waste and much more. To a great extent, the
fate of cultivation depends on the reception of different cognitive solutions. Although research
on a large scale is still ongoing and some applications are already available on the market, the
industry is still highly underserved. Farming is still at a nascent stage when it comes to handling
practical problems faced by farmers and using automated decision making and predictive
solutions to address them. Applications need to be more robust in order to explore the huge
scope of AI in agriculture (Slaughter et al., 2008). Only then, it will be able to deal with frequent
changes in external conditions and facilitate decision-making in real time. The exorbitant cost of
various cognitive solutions available in the farming sector is another significant aspect. To
ensure that technology reaches the people, solutions need to become more accessible. The
solution will be made more affordable by an open source platform, resulting in rapid adoption
and greater penetration among farmers. The technology would be useful to help high-yielding
farmers and to provide a better seasonal crop at regular intervals.
Artificial intelligence techniques are growing increasingly and can be used by CNN,
RNN or any other computer network to detect plant diseases or any unwanted weed on the field.
Greenhouse farming may give the plants a specific climate, but without human interference, it is
not possible. Wireless technology and IOT are being applied here and we can implement weather
monitoring and control without human presence using the new communication protocols and
sensors. Harvesting of fruits and crops can also be incorporated by robots which are specialized
in working round the clock for quick harvesting. The greatest threat to farming is the damage to
crops due to any form of disaster, including pest attack. Farmers lose their crops most of the time
because of the lack of proper knowledge. The technology would be useful for farmers to protect
their cultivation from any kind of attacks in this cyber era. In this direction, AI-enabled picture
recognition will be highly helpful. While AI offers immense opportunities in the application of
agriculture, there is still a lack of awareness in farms around the world of advanced high-tech
machine learning solutions.
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CONCLUSION
The much-needed reduction of human interventions in practise is agricultural monitoring.
Demand for food is approaching its high peak on a regular basis and it is very difficult to achieve
rising demand without introducing modern agricultural methods. The primary concern is the
control of agriculture, as it helps to minimise labour and increase production. In crop selection
and to assist the farmer in selecting the fertilizers, Artificial Intelligence has been implemented.
Many essential methods will guarantee better crops and proper field management for farmers.
This ultimately helps the country's overall growth, as food is the primary need of any human
being. IOT recognized its importance in helping to track the data in real time. In an intelligent
watering system, IOT is primarily used. Since efficient use of the fresh water available is
important and can be solved with the advancement in the technology and implementation of the
automation water crisis. In this modern world , traditional techniques in agriculture have minor
effects. Water shortages and floods are both the key challenges facing farmers using the
conventional approach. The advancement of agricultural automation is motivated by several
loopholes in this mechanism and the troubling need to protect agricultural property. This book
chapter reflects an idea to automate conventional agricultural practices through the use of AI,
sensors, IOT and machine learning.
REFERENCES:
1. Ahirwar S., Swarnkar R., Bhukya S, Namwade G. Application of drones in agriculture
Int. J. Curr. Microbiol. App. Sci., 8 (1) (2019), pp. 2500-2505
2. Arif C, Mizoguchi M , Setiawan B.I, Doi R. Estimation of soil moisture in paddy field
using Artificial Neural Networks International Journal of Advanced Research in Artificial
Intelligence., 1 (1) (2012), pp. 17-21
3. Encinas C, Ruiz E, Cortez J, Espinoza A. Design and implementation of a distributed
IoT system for the monitoring of water quality in aquaculture Wireless Telecommunications
Symposium (WTS)., 2017 (2017), pp. 1-7
4. FAO.. Food And Agriculture Organization Of The United Nations, 2017 The State Of
Food And Agriculture Leveraging Food Systems For Inclusive Rural Transformation 978-92-5-
109873-8 (2017), Pp. 1-181
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5. Ferguson R.B, Shapiro C.A, Hergert G.W, Kranz W.L, Klocke N.L, Krull D.H.
Nitrogen and Irrigation Management Practices to Minimize Nitrate Leaching from Irrigated Corn
Jpa, 4 (2) (1991), p. 186
6. Gliever C, Slaughter D.C. Crop verses weed recognition with artificial neural networks
ASAE paper., 01-3104 (2001) (2001), pp. 1-12
7. Hunt E.R, Cavigelli M , Daughtry C.S.T, Mcmurtrey J, Walthall C.L. Evaluation of
digital photography from model aircraft for remote sensing of crop biomass and nitrogen status
Precis. Agric., 6 (2005), pp. 359-378
8. Jha K, Doshi A, Patel P, Shah M,A comprehensive review on automation in agriculture
using artificial intelligence Artificial Intelligence in Agriculture., 2 (2019), pp. 1-12
9. Kim Y.J ,Evans R.G, Iversen W.M. Remote sensing and control of an irrigation system
using a distributed wireless sensor network IEEE Trans. Instrum. Meas., 57 (7) (2008), pp. 1379-
1387
10. Liakos K, Busato P, Moshou D, Pearson S, Bochtis D. Machine Learning in
Agriculture: A Review Sensors, 18 (8) (2018), p. 2674
11. Maier H.R, Dandy G.C. Neural networks for the prediction and forecasting of water
resources variables: a review of modeling issues and applications Environmental Modeling &
Software (2000), pp. 101-124
12. Panpatte D.G. Artificial Intelligence in Agriculture: An Emerging Era of Research
Intuitional Science, CANADA (2018), pp. 1-8
13. Pedersen S.M, Fountas S, Blackmore S. Agricultural robots – applications and
economic perspectives Service Robot Applications. (2008), pp. 369-382
14. Sannakki S.S, Rajpurohit V.S, Nargund V.B, Kumar A, Yallur P.S. Leaf disease grading
by machine vision and fuzzy logic Int. J. Comp. Tech. Appl., 2 (5) (2011), pp. 1709-1716
15. Shah G, Shah A .Shah M. Panacea of challenges in real-world application of big data
analytics in the healthcare sector. Journal of Data, Information and Management, 2019 - Springer
16. Shobila P, Mood V. Automated irrigation system using robotics and sensors Int. J. Sci.
Eng. Res., 3 (8) (2014), pp. 9-13
17. Sicat R.S, Carranza E.J.M, Nidumolu U.B, Fuzzy modeling of farmers' knowledge for
land suitability classification Agric. Syst., 83 (2005), pp. 49-75
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18. Slaughter D.C, Giles D.K, Downey D. Autonomous robotic weed control systems: a
review Comput. Electron. Agric., 61 (1) (2008), pp. 63-78
19. Song H, He Y .Crop nutrition diagnosis expert system based on artificial neural networks
Third International Conference on Information Technology and Applications (ICITA'05),
Sydney, NSW, 2005, 1 (2005), pp. 357-362
20. Wang Y, Huang L,Wu J, Xu H.Wireless Sensor Networks for Intensive Irrigated
Agriculture, Consumer Communications and Networking Conference, 2007. CCNC 2007. 4th
IEEE (Jan. 2007), pp. 197-201
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GREEN CLOUD COMPUTING AND HEALTH CARE

Sylvia Grace J
Research Scholar, Department of CSE, Sathyabama Institute of Science Technology, Chennai
119 &
Assistant Professor, Department of CSE, K.C.G College of Technology, Chennai-97
Email: ssylviagrace@gmail.com
Cloud computing is the sole strength of ‘Information and Communication Technology

(ICT)’.Simple examples of cloud computing services are Google Docs and Email services.
Making use of these apps we can readily access documents or mail anytime and anywhere in the
globe. Cloud computing not only enables us to share the information anytime, anywhere in the
world but also from one platform to another platform. Even if it is not stored in our Personal
Computers (PC) it is readily available any time, since it is stored in a Cloud, or remote location.
The Cloud computing greatly reduces the load on servers, and increases the efficiency of IT field
as well. It centrally integrates and unifies computing standards. Any increased standards or new
quick implementation can be immediately deployed on the Cloud. The energy efficiency of ICT
has become a major issue with the growing demand of Cloud Computing. There is an
exponential growth in Cloud energy consumption and CO2 emission of Cloud infrastructure.
It has created new challenges in the environmental conditions. It has become a key
environmental concern. The advent of Green Cloud Computing builds innovative technology to
improve energy efficiency and to reduce carbon footprints and e-Waste in an optimal way.
Innovation has developed so quickly with logical progression over the world in almost all fields
including the medical field. Hence there is a need to periodically update the clinical framework
to meet their better needs.With the approach of Cloud computing the doctors can preserve their
own patient’s data related to their basic ailment, diagnosis, treatment and modern issues that
staged.This built in infrastructure enables any doctors to perform investigation and to prescribe
better remedial treatments. With the availability of high-end technology a doctor can store
information on Cloud to share to the world of medical professionals. A doctor can store
information of complex or multifactorial diseases among patients. As patients move through
different stages of recovery or deterioration, treatment also moves with them as per the stage,
changing therapeutic strategies and with the condition of the patients.
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TRANSGENIC ANIMALS
Nabiha zainab
B.Sc Biotechnology,
nabiha.zainab@gmail.com
A transgenic animal is one whose genome has been altered by the transfer of a gene or
genes from another breed or species. The animals are genetically modified for a variety of
purposes including producing drugs, increase livestock, make animals disease resistant, and
research.
Advantages:
• Transgenic animals can be specifically designed to allow the study of how genes are
regulated and how they affect the normal functioning of the body and its development.
• Many transgenic animals are designed to increase the understanding of that how genes
contribute to development of diseases and how gene therapy can cure diseases.
• The animals are also made to serve as models for human diseases, so that the research
new treatments and medicines is made possible.
• The transgenic animals are also produced for increasing livestock; the transgenic animals
produce more nutritious milk, high quality meat, eggs, etc.
• Transgenic animals are also used for vaccine testing.
Disadvantages:
• Transgenic animal project is very expensive
• Generation of transgenic animal requires high maintenance.
• There may be high mortality rate and other harmful effects on animals due to failed
attempts.
• Large number of recipients are required for embryo transfer due to low transgenesis rate.
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PRODUCTION OF HUMAN INSULIN FROM rDNA TECHNOLOGY

Nabiha zainab
B.Sc Biotechnology,
The production of human insulin from recombinant DNA technology is one of the
biggest and most beneficial advancements in the field of biological sciences, as it has facilitated
mass production of insulin at affordable cost.
WHAT IS INSULIN?
Insulin is a hormone produced in the pancreas by the islets of Langerhans, which
regulates the amount of glucose in the blood. The lack of insulin causes a form of
diabetes.Insulin helps control blood glucose levels by signalling the liver and muscle and fat
cells to take in glucose from the blood. Insulin therefore helps cells to take in glucose to be used
for energy. If the body has sufficient energy, itsignals the liver to take up glucose and store it as
glycogen.
DIABETES:
Diabetes mellitus, commonly known as diabetes, is a metabolic disease that causes high
blood sugar. The hormone insulin moves sugar from the blood into your cells to be stored or
used for energy.
With diabetes, your body either doesn’t make enough insulin or can’t effectively use the
insulin it does make. Patients with type 1 diabetes require regular dosage of insulin.
Type 1diabetes is an autoimmune disease. The immune system attacks and destroys cells in the
pancreas, where insulin is made. About 10% of the people with diabetes have type1 diabetes.
The symptoms of type1 diabetes are increased hunger, increased thirst, frequent urination,
tiredness, etc.
PRODUCTION ON INSULIN FROM rDNA:
• At first a suitable vector (plasmid) is isolated from E. coli and then it is cut open by
restriction endonuclease enzyme.
• The gene of interest (ie. Insulin coding gene) is isolated from β-cell and inserted in
opened plasmid.
• Plasmid and gene of interest are recombined together by DNA ligase enzyme
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• This recombined plasmid is inserted into suitable host cell (ie E. coli) and now this
recombined host cell starts producing insulin hormone.
• The insulin is extracted and purified.
• The insulin is then injected into patients.
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GENOME EDITING
Nabiha zainab
B.Sc Biotechnology,
Genome editing is a recent advancement in the field of biotechnology. It is the deliberate

alteration of a selected DNA sequence in a living cell.
A strand of DNA is cut at a specific point and then the naturally existing cellular repair
mechanisms, then fix the broken DNA strands. The way they are repaired can affect gene
function and new DNA sequences can be delivered when the DNA is cut and act as templates for
generating an altered sequence. Genome editing techniques can be used to delete sections of
DNA or alter how a gene functions.
The plants and organisms modified through genome editing are called genetically
modified plants and animals.
How does genome editing work?
• Genome editing uses a type of enzyme called ‘engineered nuclease’ which cuts the DNA
in a specific sequence.
• After cutting the DNA in a specific place, the cell will naturally repair the cut.
• We can manipulate the repair process to make changes or edits to the DNA in that
location of the genome.
These changes include small DNA changes, removal of a section of DNA or insertion of
a foreign DNA sequence (gene).
What is genome editing used for?
For research: Genome editing can be used to change the DNA in cells or organisms to
understand their biology, how they work and to understand mutations.
To treat diseases: genome editing has been used to modify human blood cells to treat
diseases like leukaemia and AIDS. It could potentially be used to treat other diseases and genetic
disorders
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For biotechnology: genome editing has been used in many fields of biotechnology including
agriculture to genetically modify crops to improve their yields and make them resistant to pests,
diseases and drought.
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MELANIN PIGMENTATION IN PEOPLE: INTRODUCTION, TYPES,

HEREDITARILY JOB, PRODUCTION AND FUNCTION.
Shubham
Melanin is a color created in the skin of practically all creatures on Earth. Its part in
creature endurance is basic, and differs relying upon the life form. This article tends to melanin
capacity and articulation in people and different living beings, and its function in endurance.
Skin appearance is a trademark that is influenced by numerous variables. Different qualities,
sustenance, and natural elements can assume a part in the shade of skin. One of the most
remarkable parts of skin that adds to appearance is a shade known as melanin. We should
examine melanin and its capacities in people and different creatures.
ISSUES WITH ARTICULATION OF MELANIN
In people, melanin creation is significant for the counteraction of skin malignant growths,
for example, melanoma. This implies the skin gets more obscure in many people when
introduction to daylight increments. Furthermore, abundance of melanocytes can prompt the
advancement of moles in the skin. A mole, or nevus, is normally benevolent (non-compromising)
yet may get malignant with expanded daylight introduction.
OCCURRENCE OF MELANIN
Melanin is found in a few territories of the human body including:
•Skin where it gives skin shading
•Hair
•Pupils or irises of the eyes
•Stria vascularis of the internal ear
•Areas of the mind, the substantia nigra and locus coeruleus
•The medulla and zona reticularis of the adrenal organ
Sorts of melanin
A portion of the various sorts of melanin incorporate eumelanin, pheomelanin and
neuromelanin. Eumelanin is found in the hair, skin and dull regions around the areolas. It is
especially bountiful among dark populaces and gives dark and earthy colored shade to the hair,
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skin and eyes. When eumelanin is available just in limited quantities, hair might be blonde.
Pheomelanin is likewise found in the hair and skin. This sort of melanin gives pink and red hues
and is the primary color found among red-haired people. This sort of melanin isn't as defensive
against UV-radiation instigated malignancy as eumelanin.
Neuromelanin is a type of melanin found in various zones of the cerebrum and loss of this
melanin may cause a few neurological issues.
ACCOMPLISHES MORE MELANIN MEAN MORE OBSCURE SKIN?
In view of melanin levels in the skin, skin appearances are comprehensively delegated
light, medium and dim. Higher the melanin color, more obscure the skin. What Causes Expanded
Melanin Creation?
UV-A beams from the daylight infiltrates into the lower layers of the epidermis and
trigger the melanocytes to deliver more melanin. Melanin is delivered as a safeguard component.
Daylight is the significant reason for expanded melanin creation. Different variables incorporate
hormonal uneven characters, maturing and irritation measures.
PRODUCTION AND FUNCTION
Melanin is a color that is created by cells known as melanocytes in the skin of most
creatures, including people. This color comes in various shades, contingent upon the hereditary
cosmetics of the person. Melanin comes in two fundamental structures and can extend from
yellowish-red to dull earthy colored. Eumelanin is the most well-known type of melanin and is
caramel in shading. The other essential structure is called pheomelanin, which produces rosy
earthy colored shading that is frequently connected with spots and red hair. The creation of
melanin in the individual is controlled by a few variables.
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DNA REPAIR IN MAMMALIAN CELLS ON EARTH

Shubham
Life forms are for all time presented to endogenous and exogenous operators that harm
DNA. If not fixed, such harm can bring about changes, sicknesses and cell demise. The cell
reactions to DNA harm incorporate cycles that manage its outcomes (for example resilience and
apoptosis) just as immediate adjustment of the harm by DNA fix systems, which may require
enactment of checkpoint pathways. There are different types of DNA harm, for example, base
alterations, strand breaks, crosslinks and confuses. There are likewise various DNA fix
pathways.
Each fix pathway is coordinated to explicit kinds of harm, and a given sort of harm can
be focused by a few pathways. Significant DNA fix pathways are bungle fix (MMR), nucleotide
extraction fix (NER), base extraction fix (BER), homologous recombinational fix (HR), and non-
homologous end joining (NHEJ). These pathways each require various proteins. On the other
hand, O-alkylated bases, for example, O6-methylguanine can be fixed by the activity of a
solitary protein, O6-methylguanine-DNA methyltransferase (MGMT).
MGMT eliminates the alkyl bunch in a self destruction response by move to one of its
cysteine buildups. Photolyases can part covalent obligations of pyrimidine dimers created by UV
radiation.
They tie to an UV injury in a light-free cycle, yet require light (350-450 nm) as a fuel hotspot for
fix. Another NER-free pathway that can eliminate UV-incited harm, UVER, is available in just a
couple of life forms, for example, the yeast Schizosaccharomycespombe. A key factor in UVER
is the endonuclease Uve1/UVDE, which cuts 5′ of different kinds of harm. Late work has
revealed novel pathways, for example, record coupled BER, break-actuated replication, and
nucleotide entry point fix just as interconnections between known pathways. For
straightforwardness, we don't consider these here. Albeit most fix proteins are normally
homologous between living beings, their assignments are regularly extraordinary. Here we by
and large utilize the names of human proteins.
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NON-HOMOLOGOUS END JOINING (NHEJ)

NHEJ is started by official of Ku70-Ku80 dimers to the DNA closes. In higher
eukaryotes the DNA protein kinase synergist subunit (DNA-PKcs) is in this way enrolled. DSBs
that are not appropriate for ligation might be handled by MRE11-RAD50-NBS1 and different
nucleases, for example, FEN1. Moreover, a DNA polymerase might be required. At long last, the
DNA closes are rejoined by XRCC4-DNA ligase IV.
Deficient fix of DSBs can bring about chromosomal precariousness, which is portrayed
by revisions and loss of chromosomes. Various human conditions, for example, Ataxia
telangiectasia (AT) and related issues, Nijmegen breakage disorder (NBS), just as bosom and
ovarian malignant growth brought about by transformation of BRCA1 or BRCA2, are related
with surrenders in DSB fix. Notwithstanding, these conditions are a result of imperfections in
guideline of DSB fix (for example in checkpoint enactment) as opposed to because of an
immediate inactivation of HR or NHEJ.
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HEALTH BENEFITS OF EATING TRADITIONAL FOODS

INTRODUCTION:
Traditional are foods and dishes that are passed on through generations or which have
been consumed for many generations. They are traditional in nature, and may have a historic
precedent in a national dish, regional cuisine or local cuisine.
Benefits of Traditional Foods:
• Less calories - helpful for weight control.
• Less saturated fat -- better for the heart.
• More lean meats and fish.
• More iron -- better for muscles and blood.
• More zinc -- better for wound healing and fighting infection.
• More Vitamin A -- better for vision and fighting disease.
• More calcium -- better for strong bones and teeth.
TYPES OF TRADITIONAL FOODS:
Millet rice: It is rich in minerals like calcium, copper, iron, magnesium, phosphorus,
potassium, and selenium as well as essential vitamins like folate, pantothenic acid, niacin,
riboflavin, and Vitamins B6, C, E, and K. Many of the most powerful health benefits millet has
to offer are related to its fiber content.
For those who are health conscious and are wary about what they eat, experts suggest that
millets should be a part of their daily regular diet. Millets are nutritious, non-glutinous (non-
sticky) and are not acid-forming foods, thus making them very easy to digest.Mar 27, 2012.
Millets are high in nutrition and dietary fibre. They serve as good source of protein,
micronutrients and phytochemicals. The millets contain 7-12% protein, 2-5% fat, 65-75%
carbohydrates and 15-20% dietary fibre. The essential amino acid profile of the millet protein is
better than various cereals such as maize.
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MICE WITH DIABETES SUCCESSFULLY TREATED WITH ELECTROMAGNETIC

FIELDS
R.Jumail Ahamed
III B.Sc BIOCHEMISTRY
SACRED HEART COLLEGE (AUTONOMOUS), Tirupattur.
According to the Centres for Disease Control and Prevention (CDC), more than
34million people — approximately 1 in 10 — in the United States have diabetes. Of these
individuals, the vast majority have type 2 diabetes. Type 2 diabetes occurs when a person’s cells
do not react to the hormone insulin properly. Insulin, which the pancreas produces, mediates the
ability of a person’s cells to receive blood sugar.In this situation, a person’s body can tell that
their cells are not receiving blood sugar properly, and the pancreas produces more insulin in
response. At a certain point, the pancreas cannot meet the insulin demand, and, as a consequence,
blood sugar levels increase.
The CDC highlight that high blood sugar levels can cause various serious health
conditions, including vision loss, kidney disease, and heart disease. According to the National
Institute of Diabetes and Digestive and Kidney Diseases, key approaches to treating type 2
diabetes include eating more healthful diet and being more physically active.
There are also many medications that can help a person manage the symptoms of
diabetes. However, adherence to type 2 diabetes treatment is relatively low. Research has
suggested that at least 45% of people with type 2 diabetes are unable to control their blood sugar
levels effectively.
A range of factors may contribute to a person’s ability to keep diabetes symptoms in
check, including the perceived difficulties around accessing and taking medications. Exposure to
Electromagnetic Fields (EMFs) for relatively short periods reduces blood sugar and normalizes
the body's response to insulin. The new study indicates that EMFs alter the balance of oxidants
and antioxidants in the liver, improving the body's response to insulin. This effect is mediated by
small reactive molecules that seem to function as "magnetic antennae."
“It was really odd because normally these animals have high blood sugar and type 2
diabetes, but all of the animals exposed to [electromagnetic fields] showed normal blood sugar
levels. Many animals sense the Earth's electromagnetic field and use it to orient themselves as
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well as for navigation. This literature pointed to a quantum biological phenomenon whereby
EMFs may interact with specific molecules.
There are molecules in our bodies that are thought to act like tiny magnetic antenna,
enabling a biological response to EMFs. Some of these molecules are oxidants, which are studied
in redox biology, to help probe the action of an oxidant molecule called superoxide, which is
known to play a role in type 2 diabetes.
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SYNTHESIS OF CERIUM OXIDE NANOPARTICLES FROMDIFFERENT

MEDICINAL PLANTS AND THEIR ACTIVITIES AND ITS
MEDICINAL APPLICATION -A REVIEW
K.Sheela
RESEARCH SCHOLAR of BIOCHEMISTRY
Sacred Heart College (Autonomous), Tirupattur, Tamil Nadu, India
ABSTRACT
A cerium oxide nanoparticle (nanoceria) has a wide range of applications in different
fields, especially biomedical division. As a major of concern, it has a major impact on the human
health and environment. The aim of this review is to address the different medicinal plants to
synthesis of nanoceria by using Traditional, Chemical and Green Synthesis Methods and
Characterization and their medicinal application of nanoceria for antibacterial, Toxicity,
Antiradical activity and also Fe2+ Chelating activity. Finally it is very cost-effective and simple.
We have exclusively discussed on the activity of nanoceria exposed to targeted Pharmaceuticals
fields and its medicinal application.
KEYWORDS: Antioxidant; cerium oxide nanoparticles; Green Synthesis; Toxicity;
Antibacterial activity; Antioxidant activity
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STUDY ON STRENGTH PROPERTIES OF CEMENT CONCRETE PAVER BLOCKS

MADE WITH SILICA FUME – A REVIEW
S.GUNASEKAR* & Dr.N.RAMESH
Assistant Professor, Department of Civil Engineering, K.S.Rangasamy College of Technology,
Tiruchengode – 637215, Tamil Nadu, India.
Professor, Department Of Civil Engineering, K.S.Rangasamy College of Technology,
Tiruchengode – 637215, Tamil Nadu, India.
*Corresponding author E-Mail ID: guna.skr87@gmail.com
ABSTRACT
This research study presents an experimental investigation on the effect of silica fume
(SF) on the strength properties of cement concrete paver block. In this current investigation, SF
was used as replacement material for cement. SF is partially replaces the cement by 5%, 10%,
15% and 20% by weight. Aging times were 28 days for compression strength.
Keywords: Paver blocks, Cement replacement, Silica fume, Strength Property Study.
INTRODUCTION
Concrete is the most common construction material prepared using cement, sand, fresh
coarse aggregates and water. With fast industrialization, infrastructure development and increase
in the population leads to huge construction activities. Due to this various construction activities
the utilization of natural resources are increased which in turn results in escalating the cost of
construction materials. Silica fume is a by-product of electric arc furnace reduction of quartz into
silicon and ferrosilicon alloy used in the electronics industry. Silica fume is replaced to Portland
cement in cement concrete paver block to improve its properties, in particular its compressive
strength.
Cement Concrete Block Paver Blocks (CCPB) has been extensively used in many
countries for quite some time as a specialized problem -solving technique for providing
pavement in areas where conventional type of construction are less durable due to many
operational and environmental constraints.ICBP technology have been introduced in India in
construction, a decade ago for specific requirement namely footpaths, parking areas etc.
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OBJECTIVES
 The main objective of this project is to produce cement concrete paver block with satisfying
all the strength properties.
 Silica fumes also a waste and used as replacement materials for cement in concrete paver
blocks.
LITERATURE STUDY
GENERAL
This chapter describes the review of literature on cement concrete paver block using
silica fume and briefly narrates the investigations carried by various strength properties on paver
blocks in eco-friendly manner.
LITERATURE REVIEW
ChukwudiOnyeakpa (2014) in this paper, a dual mould interlocking block machine with
compaction effort was effectively constructed for interlocking block production. The materials
were sampled from kilometer 69 Maiduguri road and Bama road which was tested in the
laboratory to ensure their properties for production of interlocking blocks. Therefore, the
interlocking blocks attained ample compressive strength and can resist impact of bullets and
other sharp objects using available local materials such as lateritic clay, river sand, gravel,
Portland cement and water which cannot be afforded by the hollow blocks. Ashish V Talati
(2014) in this the author studied from the test result it is observed that for various proportions i.e.
10 % , 20% , 30% & 40% of Fly ash compressive strength of paving block is little less than the
compressive strength than conventional block. Only little economy can be achieved & waste may
utilized save the land from dumping of non-degradable material. If we talk about other two waste
material silica fume & Abrasive waste, silica fume gives less compressive strength & Abrasive
waste gives more compressive strength than the conventional block. These both options give
considerable change in strength.
Isa Yuksel and TurhanBilir (2007) presented the result of research aimed at studying the
possible usage of bottom ash (BA) and granulated blast furnace slag (GBFS) in production of
plain concrete elements. Sufficient number of briquettes, paving blocks and kerbs specimens
containing GBFS and BA as fine aggregate replacement were produced in laboratory. Then, a
few tests were conducted for investigating durability and mechanical properties of these
specimens. Unit weight, Compressive strength, freeze-thaw and water absorption and surface
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abrasion tests were conducted for paving blocks. Surface abrasion and flexural tensile strength
tests were conducted for kerb specimens. While compression strength was decreased slightly ,
durability showed that usage of partially fine aggregate of these industrial by – products have
more beneficial effects o durability characteristic of plain concrete elements.
Bhanumathidas &Kalidas, (2002) with their research on Indian fly ashes reported that the
increase in ground fineness by 52 % could increase the strength by 13%. Whereas, with the
increase in native fineness by 64% the strength was reported to increase by 77%. Looking in to
the result it was proposed that no considerable improvement of reactivity could be achieved on
replacement of aggregate with Kota stone waste aggregate where less water absorption, good
compressive strength and good splitting tensile strength. Following important conclusion are
drawn based on the discussion of the above test results. From the overall study we came to know
that the kota stone waste gives the satisfactory result for the compressive strength 47.39MPa.
The water absorption percentage is also satisfactory as per IS requirement with the good tensile
splitting tensile strength. Therefore Kota stone waste with the 50 to 60% replacement of the
aggregate gives the optimum results than the other proportion %used. Therefore it also helps in
management of disposal of Kota stone industry waste. Only the limitation in this study is cost
require for loading, unloading and crushing of Kota stone industry waste in small quantity is not
comparatively advisable because it does not give sufficient cost saving in manufacturing cost of
concrete paving block. From this result we can conclude that technically Kota stone waste
aggregate shall be used for manufactured of concrete paving block. Further investigation can also
be made by using fly ash as replacement of fine aggregate along with Kota stone industry waste
to further reduce the cost of paving block.
M.Nishanth premhar (2019) The Effect of silica fume (SF) various strength properties of
replacement of cement, coarse aggregate. 10% of cement weight replaced by silica fume. M40
grade of cement is used for mix proportion 0.4:1:2.10:2.60 .Coarse total ought to adjust to IS-
383-1970. Silica fume is added to improve the mechanical properties of SCC. It is a mineral
composed of ultra-fine solid, amorphous glassy spheres of silicon di-oxide. The maximum size
of coarse aggregate used is 10mm. Zone -II fine aggregate is adapted. Target mean strength of
26.6 N/mm2. Block size used 250mm×125mm×80mm. More than 20% of silica fume does not
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affect the properties of cement. Water absorption is less than 7% .result shows that 30% of silica
fume in paver block attain its maximum compressive strength.
CONCLUSIONS
From these journals we have learnt about various materials used for the manufacturing of
paver blocks. Then the properties and strength of these materials used in the journals were
studied. The waste materials like waste marbles, fly ash, coal waste, polyethylene, electroplating
waste, mineral waste, etc…. are studied for the strength properties. Various properties like
compressive strength, flexural strength, water absorption, abrasion resistance of paver blocks
made with different materials are studied from these journals.
After the completion of all the tests, it was observed that the mechanical properties of
paver blocks, cubes and cylinders were increased with the replacement of Silica Fume. The
micro structural properties like water absorption for paver blocks were also identified.
REFERENCES
1. Vireen Limbachiya ,Eshmaiel Ganjian & Peter Claisse (2016), “Strength ,durability and
leaching properties of concrete paving blocks incorporating GGBS and SF”, Journal of
Construction and building materials”, 113 (2016) 273-279.
2. K .ShyamPrakash&,Ch. HanumanthaRao (2016). “Study on compressive strength of
quarry dust as fine aggregate in concrete”. “Hindawi publishing corporation”, Volume
2016, Article ID1742769, 5Pages.
3. Abdul Rachman D jamaluddin, Muhammad Akbar Caronge&M.W.Tjaronge(2019)“
Evaluation of sustainable concrete paving blocks incorporating processed waste Tea ash.
“Science direct Case studies in construction materials”, 12(2020) e00325.
4. M.Nishanthpremhar,R.Jeyasundar,L.Muthukumar&A.Manojkumar(2019)“Experimental
investigation on concrete paver block by adding silica fume”.“International Research
Journal of Engineering and Technology (IRJET)” Volume: 06 Issue 4e-ISSN: 2395-
0056,pISSN: 2395-0072.
5. Akaash Gupta, Prof .ArchanaTiwari(2016). “Effect on mechanical properties of paver
block consist crusher stone dust as fine aggregate with inclusion of steel fiber”
“IJRASET” Volume 04 Issue X ISSN: 2321-9653.
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6. Kumar B.,Tike G.K. and Nanda P.K., (2007),’Evaluation of Properties of High Volume
Fly Ash Concrete for Pavements’, Journal of Materials in Civil
Engineering,Vol.19,No.10,pp.906-911.
7. G Navya(2014),”Experimental Investigation on Properties Concrete Paver Block”
Department of Civil Engineering, G,M.R.I.T, India, ISSN:2248-9622,Vol 4, Issue
8(version 6), PP 33-38, August 2014.
8. Mehta, P.K., (1999), ‘High Performance, High-Volume Fly Ash Concrete for Sustainable
Development’, International Workshop on Sustainable Development and Concrete
Technology, pp 3-14.
9. R.C.Yeole(2014),”Comparison of Mix Designs of Paver Blocks” Department of Civil
Engineering,GovtCollege of Engineering Aurangabad ,Maharashtra, India,ISSN 2250-
2459,ISO 9001:2008 Certified Journal, Volume 4, Issue 10, October 2014.
10. IS 7245:1974 Specification for Concrete Pavers.
ISBN: 978-81-948129-1-3 Page 556

A REVIEW ON THE BIOSYNTHESIS OF NANOPARTICLES BY MARINE

ORGANISMS
Abinaya D , Brita John C1 and Dhivya P2*
1
1
Post Graduate Student, Department of Chemistry, Nirmala College for Women,
Coimbatore, Tamil Nadu, India
2
Assistant Professor, Department of Chemistry, Nirmala College for Women,
Coimbatore, Tamil Nadu, India
2
dhivsanto@gmail.com
Abstract
In this novel world, we are searching for eco-friendly methods for the developing
process. Currently scientists from Nanotechnology and Nano sciences have contributed a lot for
the improvement. This review summarizes the bio-synthesis of nanoparticles by marine
organisms. Marine organisms contain a vast variety of bio-active compounds, which help in the
synthesis of nanoparticles.
Keywords: Nanotechnology, biosynthesis, marine,
1. Introduction
Nanotechnology implies the fabrication of particles with nano dimensions with respect to size,
shape, chemical composition and controlled diversity and potential uses for human benefits. [1]
Nanoparticles are produced by chemical and physical methods which are expensive, non-reliable
and potentially dangerous to the environment. Alternatively nanoparticles produced by biological
methods seem to gain interest as they are supposed to be inexpensive and eco-friendly.
Biosynthesis of nanomaterials is a kind of bottom up approach and mainly involves oxidation
and reduction reaction.[2] Nanotechnology has a wide range of applications mainly in the field of
biomedical applications, drug delivery system, ultra-sensitive disease detection. [3] Nanoscience a
new inter disciplinary subject depends on the fundamental properties of nano size objects. [4]
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Nanoparticle possess wondrous optical, electronic, magnetic, and catalytic properties than the
bulk material owing to their high surface area to volume ratio. [5]
In recent areas, a convergence between biological based technologies, green chemistry
and nanotechnology has taken place. The objective of this convergence is to create new materials
and manufacturing processes that reduce or eliminate the use of hazardous substances. [6] An
important aspect of nanotechnology is the synthesis of nanometer scale materials and the direct
control of particle morphology and dimensions during formation. Nanometer scales materials at
least one dimension less than 100nm and can have a wide variety of geometric shapes such as
plates, sheets, tubes, wires, and particles. Numerous studies have shown that nanometer scale
material exhibit unique chemical, physical, electronic, optical, thermal, mechanical, and
biological properties that significantly differ from their bulk scale counter parts. [7] These unique
properties result from the extremely small size, shape, and size distribution. In addition, because
of their small size, nanomaterials can act as bridge between bulk scale materials and molecular
structure.[8] In terms of composition, nanomaterial can be broadly classified in to two types
namely organic and inorganic. Organic nanomaterials are carbon based while inorganic
nanomaterials include noble metals (gold, silver), magnetic materials (iron oxide Fe 3O4), and
semiconductors such as titanium dioxide and zinc oxide.[9][10]
2. Marine bio nanotechnology
The synthesis of nanoparticles using marine resources, their mechanism and biological
activities to bring out the prospects in marine bio nanotechnology. [11][12] Marine bio
nanotechnology is an exciting and growing area of research in modern era. In recent years, wide
ranges of marine bio resources are extensively used in the field of nanoscience and
nanotechnology.[13–19] Nano structures of 1–100 nm are seen in marine resources such as
seashells, pearls and fish bones. The rough skin surface of dolphins and whales prevents
biofoulers attachment. This is due to presence of nanoridges with pore size of 0.2 μm 2on its skin.
The biological entities from marine resources have typical nanostructures such as cover of silica
and coral reefs with calcium have significant architectures that are seen in sponges and
diatoms.[20] The existing knowledge on potential applications and current information about
research on nanoparticles derived from marine resources. Origin of nanostructures in marine
resources gives a clue to explore the advantages and applications of nanoparticles in various
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aspects. Though marine resources have vast potential but little is explored. Most of the studies on
biological synthesis of nanoparticles have been restricted to terrestrial organisms. [12]
2.1 Marine Flora-based nanoparticles
Biosynthesis of nanoparticles by means of physical and chemical processes is highly
expensive. In order to reduce the inevitable expenses in downstream processing of the
synthesized nanomaterials and to increase the application of nanoparticles, the scientific
community targeted the biological organisms. Nature has devised various processes for the
synthesis of nano and micro-length scaled inorganic materials which have contributed to the
development of relatively new and largely unexplored area of research based on the biosynthesis
of nanomaterials.[21] Plants are important, safe and easily available source for nanoparticle
synthesis with broad variability of metabolites that may aid in reduction. Numbers of plant are
being currently investigated for nanoparticle synthesis for their efficacy and so many researches
has been done with plants with respect to phytochemicals. The main phytochemicals responsible
for their activity have been identified as terpenoids, flavones, ketones, aldehydes, amides and
carboxylic acids.[22] In this regard, plants and plant part extracts based biosynthesis has been
found to be cost effective and ecofriendly.[23]
Environmental conditions of marine ecosystem and characterization of marine plants are
extremely different from terrestrial ecosystem. Therefore, the marine plants might produce
different types of bioactive compounds including polyphenols, flavonoids, alkaloids and
tannins.[24-27] Particularly, the biosynthesis of nanoparticles from mangroves and mangrove
associates are very limited. Coastal plants especially mangroves and mangrove associates are the
good source of nanoparticles.[28]
2.2 Marine microorganism-based nanoparticles
Microorganisms from bacteria to fungi have been used in recent years are chosen to
synthesize nanoparticles in a non-toxic and environment friendly way. [29] Some microorganisms
can survive and grow even at high metal ion concentration due to their extraordinary resistant
capability.[30,31] Synthesis of nanoparticle using microbes offers better size control through
compartmentalization in the periplasmic space and vesicles. The rate of intracellular particle
formation and therefore the size of the nanoparticles could, to an extent, be manipulated by
controlling parameters such as pH, temperature, substrate concentration and time of exposure to
substrate.[32]
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Marine microbes play several important roles in synthesis of nano-based drugs for human
life improvement. Marine microbes have the potent to synthesize nanoparticle since the marine
microbes exist in the sea bottomover millions of years in the past for reducing the vast amount of
inorganic elements deep in the sea. Additionally, nanoparticles synthesized by microorganisms
tend to be stabilized by peptides such as phytochelatins, thus preventing aggregation. [33] These
short peptides are synthesized in response to heavy metal stress and have been implicated as a
universal mechanism to sequester metal ions in bacteria [34] and fungi.[35]
Nanotechnology involved in number of fields resulted in fulfilling the requirement of the
human beings. The DNA, RNA and protein-based applications induced by nanotechnology are
known as biomolecular nanotechnology, the medical applications such as treatment and disease
diagnosis merge put under nano medical technology. [36]
Many microorganisms are known to produce nanostructured particles with properties
similar to chemically synthesized material, some of which include the formation of magnetic
nanoparticles by magnetotactic bacteria, the production of silver nanoparticles within the
periplasmic space of Pseudomonas stutzeri and the formation of palladium nanoparticles using
sulphate reducing bacteria.[32] Intracellular silver nanoparticles synthesized by a marine
bacterium, Idiomarina sp. PR58-8 which was found to be highly silver tolerant.[37] The mangrove
derived microbes Escherichia coli, Aspergillus niger, Penicillium fellutanum and
Thraustochytrids capable of reducing the silver ions in faster rate with various antimicrobial
applications.[38-43] Similar to this marine bacteria and fungi some of the mangrove-derived yeast
species like Pichia capsulata and Rhodosporidium diobovatum also reported to have the
nanoparticles synthesizing capacity.[44,45] The marine cyanobacterium, Oscillatoria willei is
known to secrete a protein which is responsible for reduction of silver ions and stabilization of
silver nanoparticles.[46] Several researchers have proved that various microorganisms such as
bacteria, fungi, actinomycetes, cyanobacteria, yeast and viruses are used for the synthesis of
metallic nanoparticles by one-pot synthesis.[32][47,48]
2.3 Marine algae-based nanoparticles
Marine algae is widely used in food, medicine, and manufacturing industries. [49] It is a
rich source of biologically active compounds, such as polysaccharides (alginate, laminaran,
fucoidan), polyphenols, carotenoids, fiber, protein, vitamins and minerals. [50-52] The algal
phytochemicals include hydroxyl, carboxyl, and amino functional groups, which can serve both
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as effective metal-reducing agents and as capping agents to provide a robust coating on the metal
nanoparticles in a single step.[53]
2.4 Marine animals-based nanoparticles
Marine animal mediated nanoparticles synthesis is very sparse due to the wide
availability of renewable source such as marine microbes and planktons. Presence of nano
structures on shark skin gave a new opening for the advancements in marine nanotechnology in
synthesis and designing of nanomaterials used for biomedical applications. [54,55] Cod liver oil
mediated synthesis of silver nanoparticles observed with 5–10 nm dimension with high
monodispersity.[56] Dolphins and whales have rough skin surface due to the presence of
nanoridges. These ridges enclose a pore size of 0.2 μm 2which is below the size of marine fouling
organisms and hence there is no attachment of biofoulers. [54] Nano scaled structures found on
shark skin and 'brick-and-mortar' arrangement like micro-architecture on nacre (mother of pearl)
paved a way for the latest advances on production of synthetic designed materials, in particular
to be used in biomedical applications.[55,57] It is well known that fish is a predominant constituent
of human diet and one of the quality animal proteins available to million across the world. Fish
serves as a vital health food owing to its higher protein, beneficial fat and various micro
nutrients. Moreover, during the past several decades fisheries and aquaculture are subsidized to
global food security, poverty alleviation, rural livelihoods, employment and income
generation.[58]
2.5 Marine based nanoparticles on insect
Crop loss to the turn of 30% in plants caused due to the insect pests infesting several crop
plants. The use of chemical insecticides and pesticides in crop protection disturb the soil health,
water bodies and finally it affects human health.[59] The potential application and benefits of
nanotechnology are enormous. Recently, silver nanoparticles synthesized from marine bacterium
Shewanella algae to control pests.[60] Nanotechnology in agriculture plays an important role in
the slow release effects which includes pest control with increased shelf-life to various
applications in the agricultural fields. More number of nanoparticles have been developed using
marine organisms like plants, animals, microbes etc., for variety of application. But very few
findings were reported for the insect pest management. It needs more attention for crop
protection, to meet the satisfactory level of production and to increase our economic status of
country. The agricultural application of nanotechnology can suggest development of efficient
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and potential implications for overcoming the management of pests in crops. Nanoparticles can
be used in the formulations of pesticides, insecticides, insect repellents, pheromones and
fertilizers.[61]
3. Types of nanoparticles
Nanoparticles are classified in to real sort’s viz, natural and inorganic nanoparticles.
Carbon nanoparticles are known as natural nanoparticles. Magnetic nanoparticles, noble metal
nanoparticles (platinum, gold and silver) and semiconductor nanoparticles (titanium dioxide and
zinc oxide) are gathered as inorganic nanoparticles.
Inorganic nanoparticles are progressing utilized as used in drug delivery due to their
distinctive features such as ease of use, good functionality, biocompatibility, ability to targeted
specific cell and controlled release of drugs.
There are three methods for preparation of nanomaterials - Physical, Chemical and
Biological. Traditionally nanoparticles were produced only by physical and chemical methods.
Some of the common methods are ion sputtering, solvothermal synthesis, reduction and sol gel
technique. The need for biosynthesis of nanoparticles rose as the physical and chemical
processes were cost effective. Nature has devised various processes for the synthesis of nano and
micro length scaled inorganic materials which have contributed to the development of relatively
new and largely unexplored area of research based on the biosynthesis of nanomaterials. Hence
for the biosynthesis of nanoparticles plant and microbial sources are being. [62]
Biologically synthesized nanoparticles are of considerable interest in the area of biology
and medicine due to their unique particle size and shape-dependence and their physical, chemical
and biological properties.[63] Most of the previous studies employed biomolecules (proteins,
amino acids, carbohydrates and sugars), different type of whole cells of various microorganisms
(bacteria, fungi and algae), or dissimilar plant resources (roots, leaves, flowers, bark powders,
seeds, roots and fruits) for the synthesis of metal nanoparticles.[64-67] Marine organisms are rich
source of bioactive compounds with remarkable impact in the field of pharmaceutical, industrial
and biotechnological product developments. [53]
4. Metal Nanoparticles
The existence of metal nanoparticles and their ability to impact characteristic colour dates
back decades ago. Metal nanoparticles are used in catalysis [68], sensing[69, 70]
and
optoelectronics[71] due to dependence of their absorption sensitivity, electrical, medicinal,
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magnetic[72], and catalytic properties based on size, shape, and structure. Copper, zinc, gold,
magnesium, silver, and titanium nanoparticles are of particular interest because of their
antibacterial properties, applications in medicine, dental materials, water treatment, sunscreen
lotions, and coatings.[73-76] Recently metal, metal oxides, ceramic, silicate and polymer
nanoparticles have been synthesized and used in several applications. Their small size and high
surface area have improved their use in material science and have increased their demand. Their
properties are size dependent and also depend upon surrounding medium of nanoparticles. The
required properties may be obtained by changing the environment of nanoparticles. [77]
It is already known that metallic nanoparticle function as a catalyst for various kinds of
chemical reactions. The catalytic sites of metals are located at the surface. Thus metallic
nanoparticles of 1-10nm size can work as effective catalysts. With respect to all atoms in a
particle the ratio of surface atoms increases with decreasing particle size. Metallic nanoparticles
should be stabilized under the catalytic condition, when it is used as catalysts else it will easily
coagulate in solution and lead to form aggregates, which are less effective as catalysts.
Advantages of using metal nanoparticles as catalysts are the following:
a. The temperature applied to the catalyst where the metallic nanoparticles dispersed in solution
is below the boiling point of the solvent.
b. Metallic nanoparticles dispersed in solutions can be used as photo catalysts as they are
transparent to light.
c. The size and shape of metallic nanoparticles can be easily controlled.
d. Metallic nanoparticles immobilized on solid supports acts as catalysts even for the reactions
in a gaseous phase.
5. An Outlook on Literature
Abdul-raouf et al,[78] synthesized silver nanoparticles by the reduction of aqueous solutions

of silver nitrate with powdered and solvent extract of Padina pavonia (brown algae) by biogenic
method. The silver nanoparticles synthesized by Padina pavonia were found to be supportive for
medical applications.
Kathiraven et al,[79] reported the synthesis and antibacterial activity of silver nanoparticles
using Caulerpa racemosa, a marine alga which was collected from the Gulf of Mannar, South-
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east coast of India. The synthetic procedure provided an easy scale up in future for the industrial
and therapeutic needs.
Khan et al,[80] synthesized silver nanoparticles by the reduction of silver nitrate using
formaldehyde as reducing agent, triethylamine as promoting and stabilizing agent. The
genotoxicity and oxidative stress were evaluated using Label rohita. They concluded silver
nanoparticles induce damage and the molecular repair mechanisms in aquatic organisms.
Abirami et al,[81] explored the synthesis of silver nanoparticles on fresh water fish Mystus
gulio which induced toxicity of colloidal silver. Na+-K+-ATP was estimated using Level method.
Carbohydrates were estimated by phenol-sulphuric acid method. Lowry's method, Sulpho-
vaniline method was used to estimate the total lipids.
Fawcett et al,[82] investigated the biogenic synthesis of metal and metal oxide nanoparticles
by marine algae and marine plants. They reported that polysaccharides, proteins and other
bioactive chemicals found in the cell membrane of marine alga can act as both reducing and
capping agents.
Kumar et al,[83] reported the synthesis of silver nanoparticles and phytochemicals from
Sargassum tenerrimum.The synthesized silvers nanoparticles were well characterized by UV-
Vis,FT-IR, TEM and DLS. Results showed that the antibacterial activity of silver nanoparticles
was higher than phytochemicals present.
Sahayaraj et al,[84] formulated the synthesis of bio nanoparticles using marine algae Padina
pavonica. The biosynthesized silver nanoparticles were characterized using UV-Vis
spectroscopy, FT-IR, XRD, SEM and TEM. The Padina pavonica based silver nanoparticles
inhibited the growth of test pathogens. The biosynthesized silver nanoparticles showed higher
antimicrobial activity compared to the crude extract.
Vinct et al,[85] synthesized gold and silver nanoparticles using marine plants,
microorganisms and algae as source. Pharmacological studies, cure for antiviral diseases have
been evaluated. It serves as a biomedicine against drug-resistant bacteria.
Asmathunisha et al,[86] reported the synthesis of nanoparticles from marine organisms and
formulated their mechanism and biological activities. Mangroves, salt marshes and sand dune
were used in addition to marine animals such as finfish and sponges. The utility of nanoparticles
in cancer treatments, toxicity of nanoparticles synthesized by various marine extracts on cancer
cells and normal cells had studied and their mechanism of actions and elucidated.
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Singh et al,[87] reported the biosynthesis of silver nanoparticles using marine invertebrate
(polychaete) by chemical and physical method. The synthesis nanoparticles were characterized
by UV-Vis, AFM, SEM, EDX, XRD and FTIR are taken. The possible mechanism responsible
for the synthesis of silver nanoparticles was illustrated by chemical reactions, which showed
good antibacterial activity.
Basavaraja et al,[88] reported a simple biological process for synthesizing silver
nanoparticles using fungus Fusarium semitectum. The protein stabilizes silver nanoparticles
through coating of protein moiety on the silver nanoparticles. The spectroscopy techniques such
as UV-Vis XRD, TEM, FT-IR were used to characterized the nanoparticles prepared. Medicinal
applications were envisaged.
Singh et al,[89] synthesized the nanoparticles with the help of marine resources like marine
animals, mangroves and marine microbes. The biological synthesizing method does not involve
harmful solvents and reduced downstream processing steps, which shrink the cost for their
synthesis. The method served to be ecofriendly and cost effective.
Vineela et al,[90] reported the synthesis of silver nanoparticles by biological method from
fish scales of Catla catla which possessed potent antibacterial activity against the selected fish
pathogens. The nanoparticles were characterized by SEM, UV-Vis, FT-IR spectroscopy.
Antonysamy et al,[91] intended to standardize the protocol for the synthesis of silver
nanoparticles using aqueous extract of Dictyota bartayresiana J.V.Lamouroux and evaluated the
cytotoxic potentials using brine shrimp bio-assay activity and trypan blue dye by exclusion
method.
Ramkumar et al,[92] investigated seaweed-mediated synthesis of nanoparticles to overcome
the limitations involved in physical and chemical methods. It can be applied in pharmaceutical
and biomedical fields. It was further extended to synthesis the nanoparticles by physical,
chemical, biological methods.
Muthumari et al,[93] performed the isolation and characterization of collagen from
Sardinella longiceps fish scales. The unnecessary proteins were removed demineralization
process. The collagen was extracted in two methods: acid (acetic acid) and enzymatic (pepsin)
technique. The anti-larval effect of the collagen extract was determined using mosquito larvae of
Aldes aegypti.
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Leela et al,[94] described the synthesis, characterization and applications of silver

nanoparticles using aqueous extract and individual purified compounds of Seaweed
Hypneacervicornis. The applications of these nanoparticles were extended by studying their anti
microbial, antioxidant and anti-diabetic activity.
Kala et al,[95] investigated the synthesis of silver nanoparticles from brown sea weed
Turbinaria conoides. Minimum inhibition concentration (MIC) and Minimum bacterial
concentration (MBC) were performed to analyze the antimicrobial efficacy. The results
supported the hypothesis that silver nanoparticles synthesized from fucoidan can be used for
formulation of novel bactericidal products.
Babu et al,[96] synthesized silver nanoparticles using marine bacteria Bacillus species
NJYRK which was isolated from sea weed Padina gymnosphora. It is characterized with XRD,
FT-IR, EDAX, AFM and TEM. The marine bacteria synthesized silver nanoparticles had potent
antibacterial activity even in minimum concentration against UTI bacteria.
Inbakandan et al,[97] investigated biotic factors mediated biosynthesis of silver
nanoparticles using the extract of marine sponge, Acanthella elongata. The reduction of silver
precursor yielded uniform silver nanoparticles. It is then characterized by various spectroscopic
techniques.
Ponnuchamy et al,[98] conducted a short review on biosynthesis of metal nanoparticles
which is obtained from seaweeds. The morphology and the stability of the nanoparticles obtained
from seaweeds were examined for their biomedical and environmental applications.
Narendhran et al,[99] described the biogenic combination of silver and gold nanoparticles
from marine ocean weed extract. The nanoparticles are prepared by physical, chemical and
biological methods. Their vital applications in different fields include their toxicological studies
and the significance of inclining towards the ocean weed plants and their characterization.
Rheder etal,[100] describes the synthesis of silver nanoparticles using infusion of roots and
extract of the plant Althaea officinalis. After the reduction of silver nitrate with the compounds
of Althaea officinalis, physico-chemical analyzes were performed. Toxicity was evaluated
through Allium cepa assay.
Dharmaraj et al,[101] investigated the synthesis of silver nanoparticles from the extracellular
components of the marine fish gut Cephalopholis formosa associated bacteria and evaluated its
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antagonistic activity. Two bacterial strains were isolated which are used for the synthesis of
bionanoparticles from silver nitrate.
Lakshmi et al,[102] synthesized silver nanoparticles from the haemolymph of two marine
crabs, Carcinus macnas and Ocypode quadrata and determined their influence on human and
fish pathogens. The characterization of the obtained silver nanoparticles was examined using
UV-Vis and SEM techniques. This study provides the baseline information for synthesis of
colloidal particle from marine resources.
Liang et al,[103] demonstrated the preparation of a novel functional composite material
embedding silver nanoparticles onto the surface of natural eggshell membrane fibres. This was
achieved by chemical reduction of the Ag+ anchored by the functional groups. The effectiveness
of the as-prepared silver nanoparticles had been evaluated on the 4-nitrophenol reduction to 4-
aminophenol in the presence of excess borohydride. This work provides an important foresight in
the fabrication of function al hybrid nanocomposites.
Thomas et al,[104] attempted the biosynthesis of silver nanoparticles using marine bacterial
isolate, Ochrobactrum anhtropi. The synthesized silver nanoparticles were found to exhibit
potent antibacterial activity against the selected pathogens. This supports the potential
applications of the biosynthesized nanoparticles.
Bita et al,[105] investigated the silver nanoparticles synthesized using seaweed Sargassum
angustifolium produced by biological methods. Lethal concentration tests were done using
different concentration. The results reveals the mortality rates of common carp which showed an
increasing trend with increasing concentration and expire time, which indicates toxicity of
biological synthesized silver nanoparticles in high concentration for common carp.
Chung et al,[106] synthesized copper nanoparticles by mixing copper acetate solution with
leaf extract of Eclipta prostrata. In vitro anticancer studies demonstrated the cytotoxicity value
of synthesized copper nanoparticles against tested HepG2 cells. The biosynthesized copper
nanoparticles used for therapeutic application.
Chaudhary et al,[107] synthesized nickel nanoparticles by chemical reduction method using
ethylenediamine and characterized by various spectroscopic and microscopic techniques. They
demonstrated nickel nanoparticles as potential alternative novel-metal based catalysts for
Knoevenagel condensation. The synthesized nickel nanoparticles used in medical field due to
their efficient antimicrobial function.
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Helen et al,[108] synthesized nickel nanoparticles using Dioscorea (elephant yam) tuber
extract with rapid speed. It is characterized by UV-Vis, XRD, SEM and EDAX. The biologically
synthesized nanoparticles were found to be highly toxic against different multi drug resistance
bacteria pathogens.
Batoool et al,[109] investigated copper nanoparticles using Solanum lycopersicum. Chemical
reduction technique was employed with the use of copper sulphate. It was observed that by
increasing the concentration of tomato juice, which also increases the concentration of copper
nanoparticles.
Nasrollahzadeh et al,[110] synthesized via a green method by using of Plantago asiatica leaf
extract. It reduce copper ions into Cu(0) within 5 min of reaction without using any stabilizer or
surfactant agent. Copper nanoparticles were characterized by FT-IR, UV-Vis, TEM, X-ray
diffraction and the synthesized product was characterized FT-IR and H-NMR.
Gopinath et al,[111] reported the synthesis of copper nanoparticles which is achieved due to
the reduction of copper sulphate while the aqueous leaf extract of medium oleander act as a
reducing agent. It is characterized through UV-Vis spectrophotometer and FT-IR.
Ezhilarasi et al,[112] synthesized using Moringa oleifera plant extract as a fuel. The
formation of nickel oxide nanoparticles was confirmed by XRD, FT-IR, HRTEM, EDX. The bio
nanoparticles performed using Moringa oleifera plant extract showed effective cyto toxic
activity against HR-29 (colon cancer cell lives), The synthesized Bio nanoparticles coating for
biomedical and environmental applications.
Ghorbani et al,[113] investigated the synthesis of copper nanoparticles using Salmonella
typhimurium. It was achieved by addition of culture supernatant with aqueous copper nitrate
solution. The nanoparticles were characterized by DLS and SEM. The process of reduction was
extracellular which makes it an easier method for the synthesis of copper nanoparticles.
Kale et al,[114] synthesized nickel nanoparticles using leaves by reduction of nickel ions from
nickel salts.It were characterized by FT-IR, TEM, XRD and EDX.
Vasudeo et al,[115] synthesized nickel chloride and coriander leaf extract as a reducing agent
and stabilizing agent. This method has merits over other reported methods are easily available,
inexpensive process, pollution free and reaction conditions are simple.
Shad et al,[116] conducted a short review on green synthesis and antimicrobial efficacy of
copper and nickel nanoparticles. Antibacterial efficacies of these nanoparticles were also
ISBN: 978-81-948129-1-3 Page 568
explained to give a better look in understanding and expanding the knowledge accompanied with
already described other nanoparticles.
Kuchekar at al,[117] synthesized nickel nanoparticles by biosynthesis method with the help
of Olimum santum leaf extract. The properties were characterized by using various spectroscopic
techniques. Antibacterial activity of the synthesized nanoparticles was measured by zone
inhibition method. Green synthesis method is rapid convenient and less lime consuming
environmentally safe method for the synthesis of nickel nanoparticles.
Ahmed et al,[118] synthesized nickel nanoparticles using methanol extract of Conocompus
erectus leaf as reducing agent. Characterization of the green synthesized nickel nanoparticles
was performed by SEM and XRD. The usage of plant extract for the preparation of nickel
nanoparticles makes the process cost effective, non- toxic and green method.
Sudhasree et al,[119] investigated the synthesis of nickel nanoparticles using aqueous root
extract. Nanoparticles were characterized by various spectroscopic techniques. Biological
activity of the nanoparticles and its toxicity was assessed and found to possess the good
antioxidant and antibacterial potential with significant antibacterial activity and were non-toxic.
Mohindru et al,[120] synthesized copper nanoparticles using plant extract (tea leaf extract)
with water as the medium for reduction. The nanoparticles were examined using UV, TEM,
XRD and IR. Other physical properties such as magnetic susceptibility and optical properties
were also determined.
Din et al,[121] conducted a review on green synthesis of nickel and nickel oxide
nanoparticles by using plant extract, microbial extract and naturally occurring biomolecules.
They conducted that green methods for synthesis of nickel and nickel oxide nanoparticles are
better than chemical synthetic methods.
Pawar et al,[122] demonstrated the synthesis of copper nanoparticles using leaves extract of
Gloriosa superbal. The synthesized copper nanoparticles were analyzed using UV-
spectrophotometer and FT-IR. The Gloriosa superbal plant may be effectively utilized for a
production of copper nanoparticles with economically for the many pharmaceutical applications.
El-kassas et al,[123] synthesized copper nanoparticles using Corollina offinalis linnaeus and
Corallina mediterranea areschoug aqueous extract. The copper sulphate were reduced to copper
nanoparticles. It was characterized by various spectroscopic techniques. The results suggested
ISBN: 978-81-948129-1-3 Page 569

that the bioactive and allelopathic compounds derived from two algal extracts which are
responsible for the inhibitive impacts of copper nanoparticles in Lyngbya majuscula.
Sinha et al,[124] reported the copper nanoparticles from a fish scales of Label rohita. It
were characterized by FT-IR, TEM, SAED, XRD and EDAX. The method developed depicted
the dual functional ability of the fish scale extract solution. It is efficient as photocatalysts which
provides a promising application for the degradation of dyes from industrial effluents.
Goncalves et al,[125] synthesized chitosan-stabilized Ni-Fe bimetallic nanoparticles. It
were characterized using several techniques. The by-product formed during redox reaction was
analyzed. They concluded that the reductive removal of nimesulide using the 17-
BNP/CHI(suspension) system can be useful for the reduction of the toxicity intrinsic to this kind of
nitroaromatic compounds.
Usman et al,[126] synthesized copper nanoparticles in chitosan media via chemical
reaction method. The characterization was done using UV, FESEM, UV-Vis, XRD and FT-IR
studies. The result showed the formation of high purity of copper nanoparticles with chitosan
being a major player in stabilization.
Sinha et al,[127] investigated the silver nanoparticles using aqueous extract of Pithophora
oedogonia. It was characterized by various studies. The synthesized nanoparticles used for the
application of nano medicine against multidrug resistant pathogenic bacteria.
Kanold et al,[128] demonstrated the metallic nickel nanoparticles on the embryonic
development of the sea urchin Paracentrotus lividus. It was characterized by XRD, HRTEM,
EDS and also by ICP-MS. The use of metallic nickel nanoparticles in industrial applications and
contributed to the marine environment.
Sinha et al,[129] synthesized silver nanoparticles using a waste material extract (fish scale
extract) of Label rohita. It was confirmed by various spectroscopic techniques. These
nanoparticles were utilized as a catalyst for the reduction of several aromatic nitrocompounds.
This study reported a clear insight of heterogeneous catalysis even in miscellar media.
Valli et al,[130] synthesized copper nanoparticles using Cassia auriculata leaves extract.
Copper sulphate was made to reduce with aqueous solution of Cassia auriculata leaves extract.
It was characterized by UV-Vis, FT-IR spectral studies, XRD and FESEM analysis.
Ashtaputrey et al,[131] reported the green synthesis of copper nanoparticles by using
leaves extract of Murray’s koenigii. The nanoparticles were confirmed by the spectroscopic
ISBN: 978-81-948129-1-3 Page 570
studies. It is concluded that the copper nanoparticles synthesized were very stable because of
capping and stabilizing materials present in the leaves extract.
Pradhan[132] synthesized copper nanoparticles by leaf extract of Aloe vera. The formation
of copper nanoparticles done by reduction method. Characterization of the synthesized copper
nanoparticles was done using UV-Vis, EDAX and FT-IR techniques. The process scaled up
economic viability, environmentally benign and renewable.
6. Conclusion
Biosynthesis of nanoparticles has become an interesting topic in the recent years. The
mainly aims to our mother Earth safe for our future generation. The nano-sized particles can also
be used in the medical field. The medicines in the nano size have more effect than the medicine
in micro and macro sizes. This method is an eco-friendly method, which is an alternative for the
standard method. In this bio-synthesis method, the use of chemicals and solvents are less, hence
this method is not only eco-friendly but also cost-effective. The study of nano-biotechnology in
the marine organisms are in its infant stage, hence we can study a lot from marine world. Let this
review be a stepping stone for the researchers who are interested in this area.
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ISBN: 978-81-948129-1-3 Page 578

MOLECULAR DOCKING STUDIES OF BIOACTIVE COMPOUNDS FROM

CENTELLA ASIATICA WITH BETA - LACTAMASE OF PSEUDOMONAS
AERUGINOSA: A COMPUTATIONAL APPROACH
K.Raviya Gani, Sivaranjini Annamalai* , Shoba Gunasekaran*
Department of Biotechnology, Dwaraka Doss Goverdhan Doss Vaishnav College, Arumbakkam,

Chennai –600106, Tamil Nadu, India.
*Corresponding author: Dr. A. Sivaranjini and Shoba G,

E-mail: sivaranjinia@dgvaishnavcollege.edu.in or shobag@dgvaishnavcollege.edu.in
Abstract:
Centella asiatica (L.) commonly referred to as Indian pennywort or Asiatic pennywort. It is one
of the medicinal plants that have been declared to have a variety of medical effects. Plant has
reported for its potential antioxidant, antimicrobial, cytotoxic, neuroprotective and other
activities. Bioactive constituents namely the triterpenic saponin, flavanoids and other phenolic
compounds are widely exhibited for anti-microbial property. P.aeruginosa causes common
infections in immune compromised and cystic fibrosis patients. 3D structure of Beta – lactamase
was retrieved from PDB database subjected to molecular docking against bioactive compounds
from Centella asiatica. The compounds and control subjected to ADMET properties prediction.
In this study, molecular docking anlaysis of bioactive compounds from Centella asiatica and
control drug medicarpin against beta-lactamasewas performed using autodock 4.2. The docking
interactions of Catechin shows binding energy of -7.21 kcal/mol with two hydrogen bond
interactions when compared with control medicarpin which shows -8.45 Kcal/mol binding
energy with two hydrogen bonds with beta lactamase. Hence this molecular mechanism study
suggests that bioactive compounds from Centella asiatica may act as potential therapeutic agent
against P.aeruginosa infection by interacting with biofilm forming beta lactamase protein.
Keywords: Centella asiatica, beta-lactamase, Pseudomonas aeruginosa, cystic fibrosis,

molecular docking
ISBN: 978-81-948129-1-3 Page 579

SYNTHETIC TECHNIQUES IN NANOBIOLOGY

Pranjyan Dash1 Pradeep Kumar Panda2 and Sridhar Arelli3
1
Department of Chemical and Materials Engineering, Chang Gung University,
Taoyuan, 33302, Taiwan, Email: rkpanda277@gmail.com
2
Department of Chemical and Materials Engineering, Chang Gung University,
Taoyuan, 33302, Taiwan Email: pranjyandash@gmail.com
3
Department of Physics, OPJS UNIVERSITY, Churu, Rajasthan., Mail: usri89760@gmail.com
Abstract
This review is provided a detailed overview of the synthesis, properties and applications
of nanoparticles (NPs) exist in different forms. NPs are tiny materials having size ranges from 1
to 100 nm. They can be classified into different classes based on their properties, shapes or sizes.
The different groups include fullerenes, metal NPs, ceramic NPs, and polymeric NPs. NPs
possess unique physical and chemical properties due to their high surface area and nanoscale
size. Their optical properties are reported to be dependent on the size, which imparts different
colors due to absorption in the visible region. Their reactivity, toughness and other properties are
also dependent on their unique size, shape and structure. Due to these characteristics, they are
suitable candidates for various commercial and domestic applications, which include catalysis,
imaging, medical applications, energy-based research, and environmental applications. Heavy
metal NPs of lead, mercury and tin are reported to be so rigid and stable that their degradation is
not easily achievable, which can lead to much environmental toxicity.
Introduction
Nanotechnology is a known field of research since last century. Since ‘‘nanotechnology”
was presented by Nobel laureate Richard P. Feynman during his well famous 1959 lecture
‘‘There’s Plenty of Room at the Bottom” (Feynman, 1960), there have been made various
revolutionary developments in the field of nanotechnology. Nanotechnology produced materials
of various types at nanoscale level. Nanoparticles (NPs) are wide class of materials that include
particulate substances, which have one dimension less than 100 nm at least (Laurent et al., 2010).
Depending on the overall shape these materials can be 0D, 1D, 2D or 3D (Tiwari et al., 2012).
The importance of these materials realized when researchers found that size can influence the
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physiochemical properties of a substance e.g. the optical properties. A 20-nm gold (Au),
platinum (Pt), silver (Ag), and palladium (Pd) NPs have characteristic wine red color, yellowish
gray, black and dark black colors, respectively. Fig. 1 shows an example of this illustration, in
which Au NPs synthesized with different sizes. These NPs showed characteristic colors and
properties with the variation of size and shape, which can be utilized in bioimaging applications
(Dreaden et al., 2012). As Fig. 1 indicates, the color of the solution changes due to variation in
aspect ratio, nanoshell thickness and % gold concentration. The alteration of any of the above
discussed factor influences the absorption properties of the NPs and hence different absorption
colors are observed. NPs are not simple molecules itself and therefore composed of three layers
i.e. (a) The surface layer, which may be functionalized with a variety of small molecules, metal
ions, surfactants and polymers. (b) The shell layer, which is chemically different material from
the core in all aspects, and (c) The core, which is essentially the central portion of the NP and
usually refers the NP itself (Shin et al., 2016). Owing to such exceptional characteristics, these
materials got immense interest of researchers in multidisciplinary fields. Fig. 2 shows scanning
electron microscopy (SEM) and transmittance electron microscope (TEM) images of
mesoporous and nonporous methacrylate functionalized silica (MA-SiO2). Mesoporousity
imparts additional characteristics in NPs. The NPs can be employed for drug delivery (Lee et al.,
2011), chemical and biological sensing (Barrak et al., 2016), gas sensing (Mansha et al., 2016;
Rawal and Kaur, 2013; Ullah et al., 2017), CO2 capturing (Ganesh et al., 2017; Ramacharyulu et
al., 2015) and other related applications (Shaalan et al., 2016). In this review article, we provide
a general overview on the different types, synthesis methods, characterizations, properties and
applications of NPs. The last section is also provided with the future aspects and
recommendations.
Classification of NPs
NPs are broadly divided into various categories depending on their morphology, size and
chemical properties. Based on physical and chemical characteristics, some of the wellknown
classes of NPs are given as below.
2.1. Carbon-based NPs
Fullerenes and carbon nanotubes (CNTs) represent two major classes of carbon-based
NPs. Fullerenes contain nanomaterial that are made of globular hollow cage such as allotropic
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forms of carbon. They have created noteworthy commercial interest due to their electrical
conductivity, high strength, structure, electron affinity, and versatility (Astefanei et al., 2015).
These materials possess arranged pentagonal and hexagonal carbon units, while each carbon is
sp2 hybridized. Fig. 3 shows some of the well-known fullerenes consisting of C60 and C70 with
the diameter of 7.114 and 7.648 nm, respectively.
Figure 1 Color dependence of Au NPs on size and shape (Dreaden et al., 2012).
CNTs are elongated, tubular structure, 1–2 nm in diameter (Ibrahim, 2013). These can be
predicted as metallic or semiconducting reliant on their diameter telicity (Aqel et al., 2012).
These structurally resemble to graphite sheet rolling upon itself (Fig. 4). The rolled sheets can be
single, double or many walls and therefore they named as single-walled (SWNTs), double-
walled (DWNTs) or multi-walled carbon nanotubes (MWNTs), respectively. They are widely
synthesized by deposition of carbon precursors especially the atomic carbons, vaporized from
graphite by laser or by electric arc on to metal particles. Lately, they have been synthesized via
chemical vapor deposition (CVD) technique (Elliott et al., 2013). Due to their unique physical,
chemical and mechanical characteristics, these materials are not only used in pristine form but
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also in nanocomposites for many commercial applications such as fillers (Saeed and Khan, 2016,
2014), efficient gas adsorbents for environmental remediation (Ngoy et al., 2014), and as support
medium for different inorganic and organic catalysts (Mabena et al., 2011).
2.2. Metal NPs

Metal NPs are purely made of the metals precursors. Due to well-known localized surface
plasmon resonance (LSPR) characteristics, these NPs possess unique optoelectrical properties.
NPs of the alkali and noble metals i.e. Cu, Ag and Au have a broad absorption band in the visible
zone of the electromagnetic solar spectrum. The facet, size and shape controlled synthesis of
metal NPs is important in present day cutting-edge materials (Dreaden et al., 2012). Due to their
advanced optical properties, metal NPs find applications in many research areas. Gold NPs
coating is widely used for the sampling of SEM, to enhance the electronic stream, which helps in
obtaining high quality SEM images (Fig. 1). There are many other applications, which are deeply
discussed in applications section of this review.
2.3. Ceramics NPs
Ceramics NPs are inorganic nonmetallic solids, synthesized via heat and successive
cooling. They can be found in amorphous, polycrystalline, dense, porous or hollow forms
(Sigmund et al., 2006). Therefore, these NPs are getting great attention of researchers due to
their use in applications such as catalysis, photocatalysis, photodegradation of dyes, and imaging
applications. (Thomas et al., 2015).
2.4. Semiconductor NPs
Semiconductor materials possess properties between metals and nonmetals and therefore
they found various applications in the literature due to this property (Ali et al., 2017; Khan et al.,
2017a). Semiconductor NPs possess wide bandgaps and therefore showed significant alteration
in their properties with bandgap tuning. Therefore, they are very important materials in
photocatalysis, photo optics and electronic devices (Sun, 2000). As an example, variety of
semiconductor NPs are found exceptionally efficient in water splitting applications, due to their
suitable band gap and band edge positions (Hisatomi et al., 2014).
2.5. Polymeric NPs
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These are normally organic based NPs and in the literature a special term polymer
nanoparticle (PNP) collective used for it. They are mostly nanospheres or nanocapsular shaped
(Mansha et al., 2017). The former are matrix particles whose overall mass is generally solid and
the other molecules are adsorbed at the outer boundary of the spherical surface. In the latter case
the solid mass is encapsulated within the particle completely (Rao and Geckeler, 2011). The
PNPs are readily functionalize and thus find bundles of applications in the literature (Abd Ellah
and Abouelmagd, 2016; Abouelmagd et al., 2016).
Figure 2 FE-SEM micrographs of (a) nonporous MA-SiO2 NPs, (b) mesoporous

MA-SiO2 NPs. TEM images of (c) nonporous MA- SiO2 NPs and (d) mesoporous MA-
SiO2 NPs (Lee et al., 2011).
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Figure 3. Different form of Fullerenes/buck balls (A) C60 and (B) C70.
2.6. Lipid-based NPs

These NPs contain lipid moieties and effectively using in many biomedical applications.
Generally, a lipid NP is characteristically spherical with diameter ranging from 10 to 1000 nm.
Like polymeric NPs, lipid NPs possess a solid core made of lipid and a matrix contains soluble
lipophilic molecules. Surfactants or emulsifiers stabilized the external core of these NPs (Rawat
et al., 2011). Lipid nanotechnology (Mashaghi et al., 2013) is a special field, which focus the
designing and synthesis of lipid NPs for various applications such as drug carriers and delivery
(Puri et al., 2009) and RNA release in cancer therapy (Gujrati et al., 2014).
Figure 4. Rolling of graphite layer into single-walled and multi-walled CNTs.
3. Synthesis of nanoparticles
Various methods can be employed for the synthesis of NPs, but these methods are broadly
divided into two main classes i.e. (1) Bottom-up approach and (2) Top-down approach (Wang
and Xia, 2004) as shown in Scheme 1 (Iravani, 2011).
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Scheme 1 Typical synthetic methods for NPs for the (a) top-down and (b) bottom-up approaches.
These approaches further divide into various subclasses based on the operation, reaction
condition and adopted protocols.
3.1. Top-down syntheses
In this method, destructive approach is employed. Starting from larger molecule, which
decomposed into smaller units and then these units are converted into suitable NPs. Examples of
this method are grinding/milling, CVD, physical vapor deposition (PVD) and other
decomposition techniques (Iravani, 2011). This approach is used to synthesized coconut shell
(CS) NPs. The milling method was employed for this purpose and the raw CS powders were
finely milled for different interval of times, with the help of ceramic balls and a wellknown
planetary mill. They showed the effect of milling time on the overall size of the NPs through
different characterization techniques. It was determined that with the time increases the NPs
crystallite size decreases, as calculated by Scherer equation. They also realized that with each
hour increment the brownish color faded away due to size decrease of the NPs. The SEM results
were also in an agreement with the Xray pattern, which also indicated the particle size decreases
with time (Bello et al., 2015).
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Figure 5 SEM images of (a) The untreated carbon black, (b) and (c) 10 min and 1 h
ultrasonication in POM solution (Garrigue et al.,2004).
One study revealed the spherical magnetite NPs synthesis from natural iron oxide
(Fe2O3) ore by top-down destructive approach with a particle size varies from _20 to _50 nm in
the presence of organic oleic acid (Priyadarshana et al., 2015). A simple top-down route was
employed to synthesize colloidal carbon spherical particles with control size. The synthesis
technique was based on the continuous chemical adsorption of polyoxometalates (POM) on the
carbon interfacial surface. Adsorption made the carbon black aggregates into relatively smaller
spherical particles, with high dispersion capacity and narrow size distribution as shown in Fig. 5
(Garrigue et al., 2004). It also revealed from the micrographs, that the size of the carbon particles
become smaller with sonication time. A series of transition-metal dichalcogenide nanodots
(TMD-NDs) were synthesized by combination of grinding and sonication top-down techniques
from their bulk crystals. It was revealed that almost all the TMD-NDs with sizes <10 nm show
an excellent dispersion due to narrow size distribution (Zhang et al., 2015). Lately, highly
photoactive active
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Co3O4 NPs were prepared via top-down laser fragmentation, which is a top-down process. The
powerful laser irradiations generate well-uniform NPs having good oxygen vacancies (Zhou et
al., 2016). The average size of the Co3O4 was determined to be in the range of 5.8 nm } 1.1
nm.
3.2. Bottom-up syntheses
This approach is employed in reverse as NPs are formed from relatively simpler substances,
therefore this approach is also called building up approach. Examples of this case are
sedimentation and reduction techniques. It includes sol gel, green synthesis, spinning, and
biochemical synthesis. (Iravani, 2011). Mogilevsky et al. synthesized TiO2 anatase NPs with
graphene domains through this technique (Mogilevsky et al., 2014). They used alizarin and
titanium isopropoxide precursors to synthesize the photoactive composite for photocatalytic
degradation of methylene blue. Alizarin was selected as it offers strong binding capacity with
TiO2 through their axial hydroxyl terminal groups. The anatase form was confirmed by XRD
pattern. The SEM images taken for different samples with reaction scheme are provided in
scheme 2.
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Scheme 2 Synthesis of TiO2 via bottom-up technique. SEM images showing the TiO2 NPs
(Mogilevsky et al., 2014).
SEM indicates that with temperature elevation, the size of NPs also increases (Mogilevsky et al.,
2014). Well-uniform spherical shaped Au nanospheres with monocrystalline have been
synthesized via laser irradiation top-down technique (Liu et al., 2015a, 2015b). Liu et al.
selectively transform the octahedra morphology to spherical shape by controlling the laser
treatment time and other reaction parameters. Fig. 6 provides the SEM and TEM of the prepared
Au nanospheres, which showed average diameter of 75 }2.6 nm of Au nanospheres (red column
Fig. 6e) and 72 }3.1 in edge length of Au octahedra per particle (blue column Fig. 6f).
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Figure 6 SEM for Au nanospheres (a) top view, (b) tilted view, (c) TEM image of Au
nanospheres (d) SAED pattern (inset: TEM of single Au particle), (e) and (f) size distribution
spectra of spherical and octahedral Au NPs (Liu et al., 2015a, 2015b).
More recently, solvent-exchange method is used to achieve limit sized low density
lipoprotein (LDL) NPs for medical cancer drug delivery purpose by Needham et al. In this
method nucleation is the bottom approach followed by growth which is the up approach. The
LDL NPs were obtained without using phospholipid and possessed high hydrophobicity, which
is essential for drug delivery applications (Needham et al., 2016). The monodispersed spherical
bismuth (Bi) NPs were synthesized by both top-down and bottom-up approaches (Wang and Xia,
2004). These NPs have excellent colloidal properties. In the bottom-up approach bismuth acetate
was boiled within ethylene glycol, while in top-down approach the bismuth was converted into
molten form and then the molten drop was emulsified within the boiled diethylene glycol to
produce the NPs. The size of the NPs obtained by both methods was varied from 100 nm to 500
nm (Wang and Xia, 2004). The details of this study are provided in Scheme 3. Green and
biogenic bottom-up synthesis attracting many researchers due to the feasibility and less toxic
nature of processes. These processes are cost-effective and environmental friendly, where
synthesis of NPs is accomplished via biological systems such as using plant extracts. Bacteria,
yeast, fungi, Aloe vera, tamarind and even human cells are used for the synthesis of NPs. Au
NPs have been synthesis from the biomass of wheat and oat (Parveen et al., 2016) and using the
microorganism and plant extracts as reducing agent (Ahmed et al., 2016). Table 1 provides
the merits and demerits of various top-down and bottom-up techniques with general remarks
(Biswas et al., 2012).
4. Characterization of NPs
Different characterization techniques have been practiced for the analysis of various
physicochemical properties of NPs. These include techniques such as X-ray diffraction (XRD),
X-ray photoelectron spectroscopy (XPS), infrared (IR), SEM, TEM, Brunauer–Emmett–Teller
(BET), and particle size analysis.
4.1. Morphological characterizations
The morphological features of NPs always attain great interest since morphology always
influences most of the properties of the NPs. There are different characterization techniques
formorphological studies, but microscopic techniques such as polarized optical microscopy
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(POM), SEM and TEM are the most important of these. SEM technique is based on electron
scanning principle, and it provides all available information about the NPs at nanoscale level.
Wide literature is available, where people used this technique to study not only the morphology
of their nanomaterials, but also the dispersion of NPs in the bulk or matrix. The dispersion of
SWNTs in the polymer matrix poly(butylene) terephthalate (PBT) and nylon-6 revealed through
this technique (Saeed and Khan, 2016, 2014). The same group also provides POM study of their
materials, which showed star-like spherulites of the formed materials, whose size was decreased
with the incremental filling of SWNTs. The morphological features of ZnO modified metal
organic frameworks (MOFs) were studied through SEM technique, which indicates the ZnO NPs
dispersion and morphologies of MOFs at different reaction conditions (Fig. 7) (Mirzadeh and
Akhbari, 2016).
Scheme 3 (A) Bottom-up approach: A molecular precursor is disintegrated to simpler metal

atoms that grow into colloids. (B) Topdown approach: Large drops of a metal broken into
smaller drops (Wang and Xia, 2004).
4.2. Structural characterizations
The structural characteristics are of the primary importance to study the composition and
nature of bonding materials. It provides diverse information about the bulk properties of the
subject material. XRD, energy dispersive X-ray (EDX), XPS, IR, Raman, BET, and Zieta size
analyzer are the common techniques used to study structural properties of NPs. XRD is one of
the most important characterization techniques to reveal the structural properties of NPs. It gives
enough information about the crystallinity and phase of NPs. It also provides rough idea about
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the particle size through Debye Scherer formula (Khan et al., 2017b, 2017c; Ullah et al., 2017).
This technique worked well in both single and multiphase NPs identification (Emery et al.,
2016). Nevertheless, in the case of smaller NPs having size less than hundreds of atoms, the
acquisition and correct measurement of structural and other parameters may be difficult.
Moreover, NPs having more amorphous characteristics with varied inter atomic lengths can
influence the XRD diffractogram. In that case, proper comparison of the diffractograms of
bimetallic NPs with those of the corresponding monometallic NPs and their physical mixtures is
required to obtain accurate information. Comparison of computer simulated structural model of
bimetallic NPs with observed XRD spectra is the best way to get good contrast (Ingham, 2015).
EDX, which is normally fixed with field emission scanning electron miscopy (FE-SEM) or TEM
device is widely used to know about the elemental composition with a rough idea of % wt. The
electron beam focused over a single NP by SEM or TEM through the program functions, to
acquire the insight information from the NP under observation. NP comprises of constituent
elements and each of them emits characteristics energy X-rays by electron beam irradiation. The
intensity of specific X-ray is directly proportional to the concentration of the explicit element in
the particle. This technique is widely used by researchers to give support to SEM and other
techniques for the confirmation of their elements in prepared materials (Avasare et al., 2015;
Iqbal et al., 2016). The EDX technique used to determine the elemental composition of ultra-
sonochemically synthesized pseudo-flower shaped BiVO4 NPs (Khan et al., 2017b). Similarly,
by utilizing similar technique the elemental confirmation and graphene impregnation of
In2O3/graphene heterostructure NPs was carried out, which showed C, In and O as contribut-
XPS is considered to be the most sensitive technique and it is widely used to determine the exact
elemental ratio and exact bonding nature of the elements in NPs materials. It is surface sensitive
technique and can be used in depth profiling studies to know the overall composition and the
compositional variation with depth. XPS is based on the basic spectroscopic principles and
typical XPS spectrum is composed of the number of electrons on Y-axis plot versus the binding
energy (eV) of the electrons on X-axis. Each element has their own fingerprint binding energy
value and thus gives specific set of XPS peaks. The peaks correspond come from electronic
configuration, e.g., 1s, 2s, 2p, and 3s. Lykhach et al. provide a depth electron transfer study
through CeO2 supported Pt NPs using XPS technique with support to others. They determined
that per ten Pt atoms, only one electron is elated from the NPs to CeO2 support (Lykhach et al.,
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2015). The depth profile analysis was provided to study the dispersion of boron NPs (10 nm size)
during polyethylene glycol (PEG) functionalization. Ar+ ions were used at 1.4 keV and 20 nm;
depth surface etching was performed. It was revealed that the concentration of NPs increases
from 2 to 5% with depth. This provided good evidence that boron NPs are dissolved effectively
within the bulk of functionalized PEG (Oprea et al., 2015). In similar study coreshell Au/Ag
showed similar behavior through XPS depth profiling. Wang et al. quantify the NPs coating with
this technique through XPS and STEM spectroscopies by help of SESSA software (Wang et al.,
2016). Vibrational characterization of nanoparticles is normally studied via FT-IR and Raman
spectroscopies. These techniques are the most developed and feasible as compared to other
elemental analytical methods. The most important range for NPs is the fingerprint region, which
provides signature information about the material. In one study, functionalization of Pt NPs (1.7
nm mean size) and its interaction with Alumina substrate studied via FT-IR and XPS technique.
FT-IR confirms the functionalization as it showed the signature vibrational peaks of carboxylated
C–O 2033 cm _1, respectively in addition to a broader O–H peak at 3280 cm _1. The degree of
functionalization was revealed from the red shift values of FT-IR bands (Fig. 10) (Dablemont et
al., 2008). In another study, a series of 5 mol% Eu3+ – doped rare earth metal (RE) hafnium
oxide RE2Hf2O7 (where RE =Y, Pr, La, Gd, Lu and Er) (NPs) was synthesized by correlated
techniques. FT-IR and Raman spectra analysis exhibited that the La2Hf2O7:5%Eu3+ and
Pr2Hf2O7:5%Eu3+ possessed relatively ordered pyrochlore structure as compared to RE2Hf2-
O7:5%Eu3+ compositions (RE= Y, Er, and Lu), which possess disordered fluorite structure. The
stable structures were found thermodynamically stable until high temperature of 1500 _C.
However, a disordered–ordered cause instability in the latter case, and thus thermodynamically
unstable (Pokhrel et al., 2016). More recently surface enhanced Raman spectroscopy (SERS) is
evolving as vibrational conformational tool due to its signal enhanced capability via SPR
phenomenon (Muehlethaler et al., 2016). One study reported SERS technique to study the
vibrational properties with phonons modes in nanostructured and quantum dots NPS of TiO2,
ZnO and PbS. They concluded that the enhanced spectra can be attributed to the plasmonic
resonances in semiconductor systems (Ma et al., 2011).
4.3. Particle size and surface area characterization
Different techniques can be used to estimate the size of the NPs. These include SEM, TEM,
XRD, AFM, and dynamic light scattering (DLS). SEM, TEM, XRD and AFM can give better
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idea about the particle size (Kestens et al., 2016), but the zeta potential size analyzer/DLS can be
used to find the NPs size at extremely low level. In one study Sikora et al. used DLS technique to
investigate the size variation of silica NPs with absorption of proteins from serum. The results
showed that size increased with acquisition of protein layer. However, in case of agglomeration
and hydrophilicity, DLS might prove incapable of accurate measurement, so in that case we
should rely on the high-resolution technique of differential centrifugal sedimentation (DCS)
(Sikora et al., 2016). Beside DSC, nanoparticle tracking analysis (NTA) is relatively newer and
special technique, which can be helpful in case of biological systems such as proteins, and DNA.
In NTA method, we can visualize and analyze the NPs in liquids media that relates the Brownian
motion rate to particle size. This technique allows us to find the size distribution profile of NPs
with diameter ranging from 10 to 1000 nm in a liquid medium (Filipe et al., 2010). This
technique produced some good results as compared to DLS and found to be very precise for
sizing monodisperse as well as polydisperse samples, with substantially better peak resolution.
Gross et al. detected the particle size and concentration of different sized NPs in suspensions of
polymer and protein samples and provided an overview on the effect of experimental and data
evaluation parameters (Gross et al., 2016).Large surface area of nanomaterials offers great room
for various applications and BET is the best technique to determine the surface area of NPs
materials. This technique is based on adsorption and desorption principle and Brunauer–Emme
tt–Teller (BET) theorem. Normally nitrogen gas is used for this purpose. BET produces four
types of isotherm specifically, which are labeled as Type-I, Type-II, Type-III and Type-IV
(Fagerlund, 1973). The fresh 7Cu3Ce/ZSM-5 showed typicalfeatures of Type-I isotherm obtain
from nitrogen adsorption/ desorption. It was discovered that N2 adsorption volume is
progressively increased with relative pressure until certain limit signifying the availability of
pores. The BET specific surface area for this material was 133–144 m2/g, while the total pore
volume was 0.112–0.185 cm3/g. But after sulphidation process, the BET surface area reduced to
110 m2/mg and the pore volume decreased to 0.096 cm3/g, respectively (Liu et al., 2016).
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Figure 7. SEM images of ZnO modified MOFs at different temperatures (Mirzadeh and
Akhbari, 2016).
Optical characterizations
Optical properties are of great concerned in photocatalytic applications and

therefore, photo-chemists acquired good knowledge of this technique to reveal
the mechanism of their photochemical processes. These characterizations are
based on the famous beer-lambert law and basic light principles (Swinehart,
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1962). These techniques give information about the absorption, reflectance,

luminescence and phosphorescence properties of NPs. It is widely known that
NPs especially metallic and semiconductor NPs possess different colors and
therefore, best harmonized for photo-related applications. So, it is always
interesting to know the value of absorption and reflectance of these materials to
understand the basic mechanism for each application. Ultraviolet–visible (UV–
Vis), photoluminescence (PL) and the null ellipsometer are the well-known
optical instruments, which can be used to study the optical properties of NPs
materials.
The UV/vis- diffuse reflectance spectrometer (DRS) is a fully equipped device

which can be used to measure the optical absorption, transmittance and
reflectance. The former two are supplementary to each other while the latter
(DRS) is a special technique use for sold samples mostly. The method is excep-
tionally acceptable for the determination of bandgaps of NPs and other
nanomaterials. Bandgap of materials is very important to conclude about the
photoactivity and conductance of the material. The carbon nanodot-carbon
nitride (C3N4) was found to be a metal free water splitting photocatalyst. The
photo ability of this material is directly correlated to the bandgap value of 2.74–
2.77 eV, which was calculated using UV–Vis spectroscopy (Liu et al., 2015a,
2015b). Similarly, this technique also use to see the absorption shift in case of
doping, composite formation or heterostructure NPs materials. Peng et al.
synthesis MMT, LaFeO3 and LaFeO3/MMT nanocomposites and studied
variation in their electromagnetic radia- tions absorption through UV–vis DRS
to reconnoiter their optical characteristics. The strong red shift observed in case
of nanocomposite as compared to pristine MMT and LaFeO3 NPs. LaFeO3 and
LaFeO3/MMT displayed rather broad absorption band from 400 to 620 nm,
showing decrease in their bandgap. This property makes these catalysts
considerable for solar light driven photocatalysis (Peng et al., 2016).
In addition to UV, PL also considers valuable technique to study the optical
properties of the photoactive NPs and other nanomaterials. This technique offers
additional information about the absorption or emission capacity of the
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materials and their effect on the overall excitation time of photoexcitons. Thus, it
provides significant information about the charge recombination and half-life of
the excited materials in their conductance band, which are useful for all photo
related and imaging applications. The PL spectrum can be recorded as emission
or absorbance depending on the nature of study. Fig. 11 shows a typical PL
spectrum of pristine and modified ZnO NPs. It is evident from this figure that
pristine ZnO NPs show high PL intensity as compared to CdS modified ZnO
NPs. The gold embedded CdS/Au/ZnO composite shows the lowest intensity.
This quenching from pure ZnO to CdS/ Au/ZnO can be attributed to the
decrease in the rate of charge recombination and larger lifetime of photoexcitons
in the latter case (Yu et al., 2013). In addition, this technique is successfully used
to determine the thickness of layer (Lin et al., 2015), doping quantity of (Gupta
et al., 2013; Pal et al., 2012) material and defects/oxygen vacancies
determination (Torchynska et al., 2016) of NPs.
Figure 8 TEM images of different form of gold NPs, synthesized by different

techniques (Khlebtsov and Dykman, 2011, 2010a, 2010b).
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Figure 9 SEM (a–c, h), TEM (d–f), XRD patterns (g) and HRTEM (i) images of double,
triple and quadruple Co3O4 hollow shells (Wang et al., 2013).
1. Physicochemical properties of NPs
As discussed earlier, various physicochemical properties such as large surface

area, mechanically strong, optically active and chemically reactive make NPs
unique and suitable appli- cants for various applications. Some of their important
properties are discuss in the following section.
Electronic and optical properties
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The optical and electronic properties of NPs are inter- dependent to greater
extent. For instance, noble metals NPs have size dependent optical properties and
exhibit a strong UV–visible extinction band that is not present in the spectrum of
the bulk metal. This excitation band results when the incident photon frequency
is constant with the collective excitation of the conduction electrons and is known
as the localized surface plasma resonance (LSPR). LSPR excitation results in the
wavelength selection absorption with extremely large molar excitation coefficient
resonance Ray light scattering with efficiency equivalent to that of ten
fluorophores and enhanced local electromagnetic fields near the surface of NPs
that enhanced spectroscopies. It is well established that the peak wavelength of the
LSPR spectrum is dependent upon the size, shape and interparticle spacing of the
NPs as well as its own dielectric properties and those of its local environment
including the substrate, solvents and adsorbates (Eustis and El- Sayed, 2006).
Gold colloidal NPs are accountable for the rusty colors seen in blemished glass
door/windows, while Ag NPs are typically yellow. Actually, the free electrons on
the surface in these NPs (d electrons in Ag and gold) are freely trans- portable
through the nanomaterial. The mean free path for Ag and gold is 50 nm, which is
more than the NPs size of these materials. Thus, no scattering is expected from the
bulk, upon light interaction, instead they set into a standing resonance
conditions, which is responsible for LSPR in these NPs (Fig. 12) (Khlebtsov and
Dykman, 2010a, 2010b).
Magnetic properties
Magnetic NPs are of great curiosity for investigators from an eclectic range of
disciplines, which include heterogenous and homogenous catalysis, biomedicine,
magnetic fluids, data storage magnetic resonance imaging (MRI), and
environmental remediation such as water decontamination. The literature
revealed that NPs perform best when the size is <critical value i.e. 10–20 nm
(Reiss and Hu¨ tten, 2005). At such low scale the magnetic properties of NPs
dominated effectively, which make these particle priceless and can be used in
different applications (Faivre and Bennet, 2016; Priyadarshana et al., 2015; Reiss
ISBN: 978-81-948129-1-3 Page 599
and Hu¨ tten, 2005; Zhu et al., 1994). The uneven electronic distribution in NPs
leads to magnetic property. These properties are also dependent on the synthetic
protocol and various synthetic methods such as solvothermal (Qi et al., 2016),
co- precipitation, micro-emulsion, thermal decomposition, and flame spray
synthesis can be used for their preparation (Wu et al., 2008).
Figure 10 FTIR spectra of platinum (1.7 nm) (a) extracted from polyol, (b)
dodecanethiol coated Pt, and (c) MUDA coated Pt (Dablemont et al., 2008).
Figure 11 Photoluminescence (PL) spectra of pristine ZnO, CdS/ZnO, and CdS/Au/ZnO

measured with 270 nm excitation wavelength at normal temperature (Yu et al., 2013).
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Mechanical properties
The distinct mechanical properties of NPs allow researchers to look for
novel applications in many important fields such as tribology, surface
engineering, nanofabrication and nanoman- ufacturing. Different mechanical
parameters such as elastic modulus, hardness, stress and strain, adhesion
and friction can be surveyed to know the exact mechanical nature of NPs.
Beside these parameters surface coating, coagulation, and lubrication also
aid to mechanical properties of NPs (Guo et al., 2014) (see Scheme 4). NPs
show dissimilar mechanical properties as compared to microparticles and
their bulk materials. Moreover, in a lubricated or greased contact, the
contrast in the stiffness between NPs and the contacting external surface
controls whether the NPs are indented into the plan surface or deformed
when the pressure at contact is significantly large. This important
information could divulge how the NPs perform in the contact situation.
Decent controls over mechanical features of NPs and their interactions with
any kind of surface are vital for enlightening the surface quality and elevating
material removal. Fruitful outcomes in these fields generally need a deep
insight into the basics of the mechanical properties of NPs, such as elastic
modulus and hardness, movement law, friction and interfacial adhesion and
their size dependent characteristics (Guo et al., 2014).
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Figure 12 Graphical illustration exemplifying the localized surface plasmon (LSPR)

on nanoparticle outer surface (Khlebtsov and Dykman, 2010a, 2010b).
Scheme 4 Schematic view of the mechanical properties and their applications (Guo et
al., 2014).
ISBN: 978-81-948129-1-3 Page 602

1. Applications of NPs
Considering the unique properties discussed in Section 5, NPs can be used in
variety of applications. Some important of these are given below.
Applications in drugs and medications

Nano-sized inorganic particles of either simple or complex nature, display
unique, physical and chemical properties and represent an increasingly
important material in the development of novel nanodevices which can be used
in numerous physical, biological, biomedical and pharmaceutical applications
(Loureiro et al., 2016; Martis et al., 2012; Nikalje, 2015).
NPs have drawn increasing interest from every branch of medicine for their
ability to deliver drugs in the optimum dosage range often resulting in increased
therapeutic efficiency of the drugs, weakened side effects and improved patient
compliance (Alexis et al., 2008). Iron oxide particles such as magnetite
(Fe3O4) or its oxidized form maghemite (Fe2O3) are the most commonly
employed for biomedical applications (Ali et al., 2016). The selection of NPs for
achieving efficient contrast for biological and cell imaging applications as well
as for photo thermal therapeutic applications is based on the optical properties of
NPs. Mie theory and discrete dipole approximation method can be used to
calculate absorption and scattering efficiencies and optical resonance
wavelength for the commonly used classes of NPs i.e. Au NPs, silica-Au NPs and
Au nanorods (Jain et al., 2006). The development of hydrophilic NPs as drug
carrier has represented over the last few years an important challenge. Among
the different approaches, polyethylene oxide (PEO) and polylactic acid (PLA) NPs
have been revealed as very promising system for the intravenous administration
of drugs (Calvo et al., 1997). Superparamagnetic iron oxide NPs with appropriate
surface chemistry can be used for numerous in vivo applications such as MRI
contrast enhancement, tissue repair, and immunoassay, detoxification of
biological fluids hyperthermia, drugs delivery and cell separation. All of these
biomedical applications require that the NPs have high magnetization value, a
size smaller than 100 nm and a narrow particle size distribution (Laurent et al.,
ISBN: 978-81-948129-1-3 Page 603

2010). The detection of analytes in tissue sections can be accomplished through

antigen-antibody interactions using antibodies labeled with fluorescent dyes,
enzymes, radioactive compounds or colloidal Au (Khlebtsov and Dykman,
2010b).
Over past few decades these has been considerable interest in developing
biodegradable NPs as effective drug delivery devices (Zhang and Saltzman,
2013). Various polymers have been used in drug delivery research as they can
effectively deliver the drugs to the target site thus increases the therapeutic
benefit, while minimizing side effects. The controlled release of
pharmacologically active drugs to the precise action site at the therapeutically
optimum degree and dose regimen has been a major goal in designing such
devices.
Liposomes have been used as a potential drug carrier instead of conventional
dosage forms because of their unique advantages which include ability to
protect drugs from degradation, target to the site of action and reduce the
noxiousness and other side effects. However developmental work on liposome
drugs has been restricted due to inherent health issues such as squat
encapsulation efficiency, rapid water leakage in the commodity of blood
components and very poor storage, and stability. On the other hand, polymeric
NPs promise some critical advantages over these materials i.e. liposomes. For
instance, NPs help to increase the ratability of drugs or problems and possess
convenient controlled drug release properties. Most of the semiconductor and
metallic NPs have immense potential for cancer diagnosis and therapy on
account of their surface plasmon resonance (SPR) enhanced light scattering and
absorption. Au NPs efficiently convert the strong absorbed light into localized
heat which can be exploited for the selective laser photo thermal therapy of
cancer (Prashant et al., 2007). Beside this the antineoplastic effect of NPs is also
effectively employed to inhibit the tumor growth. The multihydroxylated
[Gd@C82(OH)22]n NPs showed antineoplastic activity with good efficiency and
lower toxicity (Chen et al., 2005). Ag NPs are being used increasingly in wound
dressings, catheters and various households’ products due to their antimicrobial
ISBN: 978-81-948129-1-3 Page 604

activity (AshaRani et al., 2009). Antimicrobial agents are extremely vital in

textile, medicine, water disinfection and food packaging. Therefore, the
antimicrobial characteristics of inorganic NPs add more potency to this
important aspect, as compared to organic compounds, which are relatively
toxic to the biological systems (Hajipour et al., 2012). These NPs are
functionalized with various groups to overcome the microbial species selectively.
TiO2, ZnO, BiVO4, Cu- and Ni-based NPs have been utilized for this purpose due
to their suitable antibacterial efficacies (Akhavan et al., 2011; Pant et al., 2013;
Qu et al., 2016; Yin et al., 2016).
Applications in manufacturing and materials

Nanocrystalline materials provide very interesting substances for material
science since their properties deviate from respective bulk material in a size
dependent manner. Manufacture NPs display physicochemical characteristics that
induce unique electrical, mechanical, optical and imaging properties that are
extremely looked-for in certain applications within the medical, commercial, and
ecological sectors (Dong et al., 2014; Ma, 2003; Todescato et al., 2016). NPs focus
on the characterization, designing and engineering of biological as well as non-
biological structures < than 100 nm, which show unique and novel functional
properties. The potential benefits of nanotechnology have been documented by
many manufacturer at high and low level and marketable products are already
being mass-produced such as microelectronics, aerospace and pharmaceutical
industries (Weiss et al., 2006). Among the nanotechnology consumer products to
date, health fitness products from the largest category, followed by the electronic
and computer category as well as home and garden category. Nanotechnology has
been touted as the next revolution in many industries including food processing
and packing. Metals NPs such as noble metals, including Au and Ag have many
colors in the visible region based on plasmon resonance, which is due to collective
oscillations of the electrons at the surface of NPs (Khlebtsov and Dykman, 2010a,
2010b; Unser et al., 2015). The resonance wavelength strong depends on size and
shape of NPs, the interparticle distance, and the dielectric property of the
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surrounding medium. The unique plasmon absorbance features of these noble

metals NPs have been exploited for a wide variety of applications including
chemical sensors and biosensors (Unser et al., 2015).
Applications in the environment
The increasing area of engineered NPs in industrial and household applications
leads to the release of such materials into the environment. Assessing the risk of
these NPs in the environment requires on understanding of their mobility,
reactivity, Eco toxicity and persistency (Ripp and Henry, 2011; Zhuang and Gentry,
2011). The engineering material applications can increase the concentration of NPs
in groundwater and soil which presents the most significant exposure avenues for
assessing environmental risks (Golobicˇ et al., 2012; Masciangioli and Zhang,
2003). Due to high surface to mass ratio natural NPs play an important role in the
solid/water partitioning of contaminants can be absorbed to the surface of NPs,
co-precipitated during the formation of natural NPs or trapped by aggregation of
NPs which had contaminants adsorbed to their surface. The interaction of
contaminants with NPs is dependent on the NPs characteristics, such as size,
composition, morphology, porosity, aggregation/disaggrega- tion and aggregate
structure. The luminophores are not safe in the environment and are protected
from the environmental oxygen when they are doped inside the silica network
(Swadeshmukul et al., 2001).
Most of environmental applications of nanotechnology fall into three categories:
1. Environmentally benign sustainable products (e.g. green chemistry or pollution

prevention).
2. Remediation of materials contaminated with hazardous substances and
3. Sensors for environmental stages (Tratnyek and Johnson, 2006).
The removal of heavy metals such as mercury, lead, thallium, cadmium and
arsenic from natural water has attracted considerable attention because of their
adverse effects on environmental and human health. Superparamagnetic iron
ISBN: 978-81-948129-1-3 Page 606
oxide NPs are an effective sorbent material for this toxic soft material. So, for no
measurements of engineered NPs in the environment have been available due to
the absence of analytical methods, able to quantify trace concentration of NPs
(Mueller and Nowack, 2008). Photodegradation by NPs is also very common
practice and many nanomaterials are utilized for this purpose. Rogozea et al. used
NiO/ZnO NPs modified silica in the tandem fashion for photodegradation
purpose. The high surface area of NPs due to very small size (<10 nm),
facilitated the efficient photodegradation reaction (Rogozea et al., 2017). The
same group has reported the synthesis of variety of NPs and reported their
optical, florescence and degradation applications (Olteanu et al., 2016a, 2016b;
Rogozea et al., 2016).
Applications in electronics
There has been growing interest in the development of printed electronics in
last few years because printed electronics offer attractive to traditional silicon
techniques and the potential for low cost, large area electronics for flexible
displays, sensors. Printed electronics with various functional inks containing
NPs such as metallic NPs, organic electronic molecules, CNTs and ceramics NPs
have been expected to flow rapidly as a mass production process for new types of
electronic equipment (Kosmala et al., 2011).
Unique structural, optical and electrical properties of one dimensional
semiconductor and metals make them the key structural block for a new
generation of electronic, sensors and photonic materials (Holzinger et al.,
2014; Millstone et al., 2010; Shaalan et al., 2016).
The good example of the synergism between scientific discovery and
technological development is the electronic industry, where discoveries of new
semiconducting materials resulted in the revolution from vacuumed tubes to
diodes and transistors, and eventually to miniature chips (Cushing et al.,
2004).
The important characteristics of NPs are facile manipulation and reversible
assembly which allow for the possibility of incorporation of NPs in electric,
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electronic or optical devices such as ‘‘bottom up” or ‘‘self-assembly” approaches

are the bench mark of nanotechnology (O’Brien et al., 2001).
Applications in energy harvesting

Recent studies warned us about the limitations and scarcity of fossil fuels
in coming years due to their nonrenewable nature. Therefore, scientists shifting
their research strategies to generate renewable energies from easily available
resources at cheap cost. They found that NPs are the best candidate for this
purpose due to their, large surface area, optical behavior and catalytic nature.
Especially in photocatalytic applications, NPs are widely used to generate energy
from photoelectrochemical (PEC) and electrochemical water splitting (Avasare
et al., 2015; Mueller and Nowack, 2008; Ning et al., 2016). Beside water
splitting, electrochemical CO2 reduction to fuels precursors, solar cells and
piezoelectric gen- erators also offered advance options to generate energy (Fang
et al., 2013; Gawande et al., 2016; Lei et al., 2015; Li et al., 2016; Nagarajan et
al., 2014; Sagadevan, 2015; Young et al., 2012; Zhou et al., 2016). NPs also use in
energy storage applications to reserve the energy into different forms at
nanoscale level (Greeley and Markovic, 2012; Liu et al., 2015a, 2015b;
Sagadevan, 2015; Wang and Su, 2014). Recently, nanogenerators are created,
which can convert the mechanical energy into electricity using piezoelectric,
which is an unconventional approach to generate energy (Wang et al.,
2015). Fig. 13 shows some energy generating devices, and uses NPs.
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Figure 13 Energy generation approaches from (A) Piezoelectrics

actuators (B) Water splitting (C) CO2 reduction
Applications in mechanical industries

As revealed from their mechanical properties through excellent young
modulus, stress and strain properties, NPs can offer many applications in
mechanical industries especially in coating, lubricants and adhesive
applications. Besides, this property can be useful to achieve mechanically
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stronger nanodevices for various purposes. Tribological properties can be

controlled at nanoscale level by embedding NPs in the metal and polymer
matrix to increase their mechanical strengths. It is because, the rolling mode of
NPs in the lubricated contact area could provide very low friction and wear. In
addition, NPs offer good sliding and delamination properties, which could also
effect in low friction and wear, and hence increase lubrication effect (Guo et al.,
2014). Coating can lead to various mechanically strong characteristics, as it
improves toughness and wear resistance. Alumina, Titania and carbon based NPs
successfully demonstrated to get the desirable mechanical properties in coatings
(Kot et al., 2016; Mallakpour and Sirous, 2015; Shao et al., 2012).
7. Toxicity of NP
Beside many industrial and medical applications, there are certain toxicities
which are associated with NPs and other nanomaterials (Bahadar et al., 2016;
Ibrahim, 2013; Khlebtsov and Dykman, 2011, 2010b) and basic knowledge is
required for these toxic effects to encounter them properly. NPs surrepti- tiously
enter the environment through water, soil, and air during various human
activities. However, the application of NPs for environmental treatment
deliberately injects or dumps engineered NPs into the soil or aquatic systems. This
has resul- tantly attracted increasing concern from all stakeholders. The
advantages of magnetic NPs such as their small size, high reactivity and great
capacity, could become potential lethal factors by inducing adverse cellular toxic
and harmful effects, unusual in micron-sized counter parts. Studies also illustrated
that NPs can enter organisms during ingestion or inhalation and can translocate
within the body to various organs and tissues where the NPs have the possibility
to exert the reactivity being toxicology effects. Although some studies have also
addressed the toxicological effects of NPs on animal cells and plant cells the
toxicological studies with magnetic NPs on plants to date are still limited. The uses
of Ag NPs in numerous consumer products lead them to their release to the
aquatic environment and become a source of dissolved Ag and thus exert toxic
effects on aquatic organisms including bacteria, algae, fish and daphnia (Navarro
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et al., 2008). The respiratory system represents an unique target for the potential
toxicity of NPs due to the fact that in addition to being the portal of entry for
inhaled particles, it also receives the entire cardiac output (Ferreira et al., 2013).
NPs are used in bio applications widely but despite the rapid progress and early
acceptance of nanobiotechnology the potential for adverse health effects due to
prolong exposure at various concentrations levels in human in the environment
has not yet been established. How- ever, the environmental impact of NPs is
expected to increase in the future. One of the NPs toxicity is the ability to organize
around the protein concentration that depends on particles size, curvature, shape
and surface characteristics charge, functionalized groups, and free energy. Due to
this binding, some particles generate adverse biological outcomes through protein
unfolding, fibrillation, thiol crosslinking, and loss of enzymatic activity. Another
paradigm is the release of toxic ions when the thermodynamic properties of
materials favor particles dissolution in a suspending medium or biological
environment (Xia et al., 2008).
NPs tend to aggregate in hard water and seawater and are greatly influenced by
the specific type of organic matter or other natural particles (colloids) present in
fresh water. The state of dispersion will alter the ecotoxicity, but many abiotic
factors that influence this, such as pH, salinity, and the presence of organic
matters remain to be systematically investigated as part of ecotoxicological
studies (Handy et al., 2008).
8. Conclusion
In this review, we presented a detail overview about NPs, their types, synthesis,
characterizations, physiochemical properties and applications. Through different
characterization techniques such as SEM, TEM and XRD, it was revealed that NPs have
size ranges from few nanometer to 500 nm. While the morphology is also controllable.
Due to their tiny size, NPs have large surface area, which make them suitable candidate for
various applications. Beside this, the optical properties are also dominant at that size,
which further increase the importance of these materials in photocatalytic applications.
Synthetic techniques can be useful to control the specific morphology, size and magnetic
properties of NPs. Though NPs are useful for many applications, but still there are some
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health hazard concerns due to their uncontrollable use and discharge to natural
environment, which should be consider for make the use of NPs more convenient and
environmental friendly.
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PHYTOCHEMICALS AND PHARMACOLOGICAL APPLICATIONS OF PLANTS

Poongothai Annadurai
Assistant Professor, PG and Research Department of Biochemistry
Sacred Heart College (Autonomous),
Tirupattur – 635601, Tamil Nadu
kumuthaannadurai@gmail.com
Medicinal plants are indispensable natural resource constituting one of the potential
sources of bioactive chemical entities for drug development. Traditional medicinal uses of plants
offer valuable clue to such drug development. It is estimated that about 60% of the world
population and 80% of the population of developing countries rely on traditional medicine for
their primary health care needs. Medicinal plants satisfy the need of millions of ethnic and
indigenous people living in tribal and rural sector of India. In recent years, despite incredible
development in allopathic medicine, majority of people throughout the globe are opting for
herbal healthcare system owing to their efficacy, safety and lesser side effects; as such, herbal
medicines are gaining polarity and acceptance than ever before. Medicinal plants related ethno
medicinal knowledge has been an important guiding principle for research in the area of new
drug development. Persistent research on plants involves a persistent search for new
phytochemical molecules with specific pharmacological efficacy and that are non-toxic and
efficacious in controlling human diseases.
Plants have provided man with all his needs in terms of shelter, clothing, food, flavours
and fragrances as not the least, medicines. Plants have formed the basis of sophisticated
Traditional Medicine systems that have been in existence for thousands of years and continue to
provide mankind with new remedies. Many indigenous medicinal plants are being discovered
every day. Medicinal plants used in traditional medicine should be collected at the right time, the
right season, and the right stage of their growth so that the constituents can be optimally
harvested. The phytochemical analysis of the medicinal plants are also important and have
commercial interest in both research institutes and pharmaceuticals companies for the
manufacturing of the new drugs for treatment of various diseases.
ISBN: 978-81-948129-1-3 Page 630

Editors: Associate Editors:
Dr. SHEKHAR R Dr. KAMAL GULATI
Dr. JAGANNATHAN S Mr. MOHIT TIWARI
Mrs. JANAKI S Ms. TRIPTI TIWARI
Mrs. ALBINO WINS J Dr. ANANTH KUMAR T
Ms. MUNEESWARI K Dr. SIVARANJINI A
Ms. AMUDHANILA P Mr. UBAITHULLA BAIG A
Published by
AIB SALIHA PUBLICATIONS

Tamil Nadu, India

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Virtual International Conference

01st November 2020

Association of Global Academicians and

The Virtual International Conference on Advancements in Nanotechnology

Hello and welcome to Virtual International Conference on Advancements in

Dr. KAMAL GULATI

Mr. MOHIT TIWARI

I am delighted to know that the Association of Global Academicians and Researchers

Dr. T. ANANTH KUMAR

I am pleased to know that the Virtual International Conference on Advances

My sincere gratitude to the members of the committe

“A WOMAN’S HEALTH IS HER CAPITAL”

MEDICINAL TREE - Couroupitaguianensis

ARTIFICIAL INTELLIGENCE IN AGRICULTURE: A MINI

GREEN CLOUD COMPUTING AND HEALTH CARE

PRODUCTION OF HUMAN INSULIN FROM rDNA

DNA REPAIR IN MAMMALIAN CELLS ON EARTH

HEALTH BENEFITS OF EATING TRADITIONAL FOODS

MICE WITH DIABETES SUCCESSFULLY TREATED WITH

PHYTOCHEMICALS AND PHARMACOLOGICAL

PLANT-BASED GREEN APPROACHES OF IRON NANOPARTICLES

KEYWORDS: Iron nanoparticles; plant; bioremediation; nanotechnology

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biocompatibility, and superparamagnetism allow the iron nanoparticles to apply bioremediation.

PLANT-BASED BIOSYNTHESIS OF IRON NANOPARTICLES

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BIOREMEDIATION OF WASTEWATER USING IRON NANOPARTICLES

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7. Hazarika D, Phukan A, Saikia E and Chetia B (2014) Phytochemical screening and

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AN ASSESSMENT OF BACTERICIDAL ACTIVITY, CYTOTOXIC EFFECT AND

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KEYWORDS: Bactericidal Activity, Cytotoxic Effect, Silver Nano-particles, Annona

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MATERIALS AND METHODS

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PREPARATION OF MICRO ORGANISM:

PREPARATION OF PLANT EXTRACTS

GREEN SYNTHESIS OF SILVER NANOPARTICLES

UV-VISIBLE SPECTROSCOPY ANALYSIS

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RESULTS AND DISCUSSION

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04 Staphylococcus aureus (Isolated) 11.9 13.4 19.7 14.2 17.6 12.5

Figure- 1: Bactericidal activity of the AgNPs of Annona reticulata

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STRUCTURAL CHARACTERIZATIONS OF NANOSTRUCTURED ZINC OXIDE

THIN FILM GROWN BY SPRAY PYROLYSIS TECHNIQUES

Zayyanu Shehu , Shi’itu Abubakar , Yusuf Sarkingobir , Niyas Ahmed.M.I

KEYWORDS: Nanostructured, Zinc Oxide, thin film, Spray pyrolysis, Techniques

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2. MATERIALS AND METHODS

2.2.1 SELECTION OF SUBSTRATE

2.2.2 CLEANING OF SUBSTRATE

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2.2.3 PREPARING PRECURSOR SOLUTION

2.2.4 DEPOSITION OF PRECURSOR SOLUTION

2.2.5 ANNEALING OF ZINC OXIDE LAYER

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