HUMAN NUTRITION

CARBOHYDRATE

Table of Contents
1. Role in nutrition and health...................................................................................................4
1.1. Nutrition......................................................................................................................4
1.2. Health..........................................................................................................................6
1.2.1. Grains...................................................................................................................6
1.2.2. Potatoes................................................................................................................7
1.2.3. Legumes...............................................................................................................8
1.2.4. Fruits....................................................................................................................8
2. Function, Quantity and Quality..............................................................................................9
2.1. Function.......................................................................................................................9
2.1.1. Functions of Monosaccharides..............................................................................10
2.1.2. Functions of Disaccharides...................................................................................11
2.1.3 Functions of Oligosaccharides...............................................................................12
2.1.4 Functions of Polysaccharides................................................................................13
2.2. Quantity.....................................................................................................................15
3. Deficiency.........................................................................................................................24
4. Metabolism, Digestion and Absorption, Energy, Interaction and More....................................26
4.1. Metabolism................................................................................................................26
4.1.1. Warburg effect and redox metabolism....................................................................26
4.1.2. Mechanism of aerobic glycolysis preference...........................................................27
4.1.3. Acidification of the microenvironment and immune dysregulation...........................29
4.1.4. Therapeutics targeting glucose metabolism............................................................31
4.2. Absorption.................................................................................................................32
4.2.1. Going to Blood Stream.........................................................................................32
4.3. Energy.......................................................................................................................32
4.3.1. Carbohydrates as a source of energy......................................................................32
4.3.2. Hydrolysis...........................................................................................................37
4.3.3. Cellular respiration..............................................................................................38
4.3.4. ATP: An Important Source of Energy for Cellular Processes...................................40
 4.4. Interaction: Carbohydrate-protein interactions...............................................................43
5. Digestion...........................................................................................................................44
5.1. Enzyme action: Digestive enzymes...............................................................................45
6. Transformation of nutrients during processing and preservation..............................................47
6.1. Browning reaction.......................................................................................................47
6.1.1. Enzymatic browning............................................................................................47
6.1.2. Non – enzymatic browning...................................................................................52
6.2. Biochemical process for food flavoring.........................................................................56
7. Control reactions occuring in food.......................................................................................57
7.1. Maillard reaction.........................................................................................................57
7.2. Caramelization............................................................................................................61
7.3. Chemical oxidation of phenols.....................................................................................62
7.4. Maderisation...............................................................................................................64
8. Technology: Protecting Our Food........................................................................................65
9. Dietary Fiber: Digestion......................................................................................................67
10. References.....................................................................................................................69

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1. Role in nutrition and health
In the average human diet, carbohydrates account for between 40% and 75% of the
total amount of energy consumed. The ability to be digested is their most crucial
quality. In contrast to carbohydrates that are not hydrolyzed by endogenous human
enzymes, those that are hydrolyzed by gastrointestinal system enzymes in humans
result in monosaccharides that are absorbed in the small intestine and enter the
pathways of carbohydrate metabolism. It can, however, undergo variable degrees of
fermentation in the large intestines.

Polyhydroxy aldehydes, ketones, alcohols, acids, their simple derivatives, and

polymers having polymeric linkages of the acetal type" may be used to define
carbohydrates.

1.1. Nutrition
Foods high in carbohydrates are an essential part of a balanced diet. Carbohydrates
provide the body with glucose, which is converted into energy for physical activity and
the maintenance of biological functions.

- The healthiest forms of carbohydrates—whole grains, vegetables, fruits, and

legumes that have not been processed or have had little processing—promote
excellent health by supplying vitamins, minerals, fiber, and a variety of significant
phytonutrients.

- White bread, pastries, sodas, and other highly processed or refined meals are
among the unhealthier sources of carbs. These foods have simple carbs, which can
make it difficult to lose weight and can increase the risk of diabetes and heart
disease.

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- The breakdown of glycemic carbohydrates provides the body with the majority of
the monosaccharide glucose that it needs. Humans need a steady supply of
glycemic carbohydrates to maintain the health of their reproductive organs, brain,
and kidney medulla. 20–25% of an adult's baseline metabolic expenditure goes
toward the brain's needs. As a result, red blood cells require about 20 g of glucose
every day. Human nourishment depends on it, and the human brain needs energy.
Human nourishment depends on it, and the human brain needs energy. To bridge
the gap between its gluconeogenic capability and daily glucose needs, the brain
needs 30 to 50 grams of dietary glycemic carbohydrates.

- Blood glucose levels are kept within a specific range in healthy people to prevent
the physiological effects of hypo- or hyperglycemia. Following the breakdown of
various dietary carbohydrates, glucose enters the bloodstream in the fed state, up to
five hours after a meal. The majority of the glucose is utilized to create polymer
glycogen reserves in the muscle and liver tissues, while some of it is quickly
oxidized to produce energy. While liver glycogen keeps blood glucose levels in
check, muscle glycogen serves as the muscles' short-term energy reserve. To
provide glycemic carbohydrates for the brain, red blood cells, and other tissues that
cannot utilize fatty acids as their energy source while fasting or after absorption,
glycogen is mobilized. Due to the restricted amount of glycogen in the body,
glycemic carbohydrates are available for only 18024 hours of fasting1 .

However, it is believed that the evolution of hominins into modern humans required
and complemented nutritional components, including carbohydrates from plants and
meat. To fulfill the increased metabolic needs of a developing brain and improved
aerobic capacity, which may have been caused by omnivorous hominins, had a
1
Hardy, K., Brand-Miller, J., Brown, K. D., Thomas, M. G., Copeland, L., & Handling Editor Daniel E.
Dykhuizen. (2015). The Importance of Dietary Carbohydrate in Human Evolution. The Quarterly Review of
Biology, 90(3), 251–268. https://doi.org/10.1086/682587

4
significant evolutionary advantage in the middle and late Pleistocene era. It has also
been suggested that the switch from a high-volume. Changing from a low-energy to a
low-volume high-energy diet has caused several significant alterations in hominids
characteristics linked to the appearance of H. erectus, like a significant increase in
height and body weight, smaller teeth, and a larger brain cause the mouth to start
breaking down starch.

Numerous different types of carbohydrates are produced by plants and act as energy
stores for structural processes. Some edible roots and tubers contain up to 80% starch
by dry weight; if left intact in the soil, they remain stable and can be picked for months
as needed. The utilization of starch-rich roots and tubers in early hominin diets is
therefore considered to be a potentially essential step in setting early australopithecines
apart from other hominins and facilitating expansion into new habitats. In this regard,
it is likely that the gradual substitution of fibrous plants with more energy-efficient
plant meals, notably starchy tubers, led to a reduction in gut size. This is true even
though evidence of plant foods seldom survives, making it challenging to rebuild their
diet. Additionally, it has been suggested that although meat may have been their
favorite diet, getting it may have required more energy than getting tubers from a
trusted and safe source2.

1.2. Health
1.2.1. Grains
Grains are common foods and a significant source of dietary carbohydrates all around
the world. Grains are the seeds of cereal grasses and other related plant families.
Whole grains with little processing keep all three parts of the seed intact. When grains
are refined, the fibrous bran, protein- and fat-rich germ, and starchy endosperm are left
2
Clemente-Suárez, V. J., Mielgo-Ayuso, J., Martín-Rodríguez, A., Ramos-Campo, D. J., Redondo-Flórez, L., &
Tornero-Aguilera, J. F. (2022). The Burden of Carbohydrates in Health and Disease. Nutrients, 14(18), 3809.
https://doi.org/10.3390/nu14183809

5
behind. In comparison to diets lacking them, whole grains produce small but
significant reductions in low density lipoprotein (LDL) cholesterol, total cholesterol,
and percentage body fat. They also improve postprandial glucose levels and glucose
homeostasis. This information comes from meta-analyses of randomized clinical trials.

Prospective cohort studies have also revealed strong negative correlations between
whole grain consumption and risk of death from any cause, as well as incidence of
type 2 diabetes, coronary heart disease, ischemic stroke, and various malignancies. On
the other hand, consuming more refined grains—especially white rice—is linked to a
higher risk of type 2 diabetes. Grains with whole kernels or that have been coarsely
ground often have a lower GI than grains that have been refined, and they also include
more fiber and phytochemicals that may have anti-inflammatory and antioxidant
qualities.

Though not thoroughly researched, whole grains and wholemeal foods' relative health
advantages over other types of whole foods with lower carbohydrate contents (such as
nuts, seeds, legumes, avocado, and olives) have not been established. In addition, the
majority of whole grains found in processed goods have been milled into flour with
finer particle size and a higher GI, while different amounts of bran and germ have been
added back in.

Therefore, whole grain foods might not have the same health advantages as intact or
barely processed whole kernel grains (such as quinoa, wheat berries, and steel cut
oats), and some whole grain foods include a lot of added sugar3.

1.2.2. Potatoes

3
Ludwig, D. S., Hu, F. B., Tappy, L., & Brand-Miller, J. (2018). Dietary carbohydrates: role of quality and
quantity in chronic disease. BMJ (Clinical research ed.), 361, k2340. https://doi.org/10.1136/bmj.k2340

6
Another important source of dietary carbohydrates is potatoes, which are the most
popular vegetable in the majority of nations. Although potatoes offer certain nutrients
(including vitamin C, potassium, and fiber), as they are normally eaten, they primarily
comprise starch with a high GI.14. Even after adjusting for body mass index and other
diabetes risk variables, increased potato consumption was linked to greater weight gain
and a higher risk of type 2 diabetes in three cohorts of US men and women. Higher
consumption of baked, boiled, or mashed potatoes as well as French fries was
independently linked to a higher risk of developing hypertension in the same cohorts.
As a result, compared to other vegetables, potatoes' health consequences are more
similar to those of refined grains3.

1.2.3. Legumes
Including whole grains, legumes including beans, peas, and lentils enhance nutritional
quality and health outcomes when incorporated into regular dietary patterns. Low GI
carbohydrates, high levels of protein, fiber, and other nutrients can be found in
legumes. A non-soy legume dietary intervention significantly reduced total and LDL
cholesterol compared to control diets, according to a meta-analysis of randomized
clinical trials. A different meta-analysis compared the highest and lowest consumption
categories and discovered a 10% decreased risk of cardiovascular disease. Increasing
the proportion of beans to white rice in a Costa Rican population was linked to
reduced levels of blood lipids and blood pressure as well as other cardiometabolic risk
factors3.

1.2.4. Fruits
Whole fruits often have a moderate to low GL and are high in fiber, vitamins,
minerals, and phytochemicals. According to prospective cohort studies, eating fruits
regularly lowers the incidence of type 2 diabetes, heart disease, cancer, and all-cause
death. In three US cohorts, more eating of whole fruits—particularly blueberries,

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grapes, and apples—is significantly linked to a decreased risk of developing diabetes,
whereas greater consumption of fruit liquids is linked to a higher risk. Fruit juices tend
to have less fiber, fewer micronutrients, and a higher GI than whole fruits, and for
these reasons, it is debatable whether or not to group whole fruits and juices together
in dietary recommendations3.

The link between food and mental health has attracted a lot of attention recently. The
scientific community has traditionally concentrated on analyzing dietary trends in
general, but it is now required to look at particular macronutrients separately2.

50 to 60 percent of a person's daily food intake should be made up of carbs. This

calorie intake therefore has the greatest impact on our body. Regarding mental health,
aberrant brain function and faulty glucose metabolism can cause a variety of
psychological and physiological diseases, with depression and anxiety being the most
prevalent. The kind of dietary carbohydrate, however, makes a difference in every
situation. According to FAO and WHO guidelines, the majority of the carbohydrates
consumed should be high in dietary fiber and have a low glycemic index2.

Humans' carbohydrate intake varies depending on their stage of life. Since then,
glucose has drawn special attention because it is the brain's preferred and main source
of energy. The majority of pediatric investigations reveal that the brain seems to be
sensitive to short-term changes in glucose availability. Researchers from Nutrients
2022, 14, 3809, 10 of 28 have shown that very young infants' spoken word memory is
improved following a glucose feed2.

2. Function, Quantity and Quality

2.1. Function

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Including carbohydrates in your diet will help you stay healthy. Complex
carbohydrates are among the healthiest options because of how little impact they have
on blood sugar levels. Unprocessed whole grains, produce, fruits, and legumes are
some of the choices. Simple carbs can be consumed in moderation, but white bread,
sodas, pastries, and other heavily processed meals are less nutrient-dense and
significantly raise blood sugar levels. Carbohydrates should make up 45% to 65% of a
healthy adult's daily diet, or roughly 200 g to 300 g per day. A gram of carbohydrates
has roughly 4 kcal (or 17 kJ) in it. Another crucial carbohydrate is fiber. Since fiber
has been shown to lower the risk of coronary heart disease, strokes, and other diseases,
healthy persons should consume around 30 g of fiber each day.

A glycemic index is a tool used to track carbohydrates and their individual effects on
blood sugar. This scale ranks carbohydrates from 0 to 100 based on how rapidly the
rise in blood glucose occurs upon consumption. Low glycemic foods (55 or less)
produce a gradual increase in blood sugar. These foods include steel-cut oatmeal, oat
bran, muesli, sweet potatoes, peas, legumes, most fruits, and non-starchy vegetables.
Medium glycemic foods (56 to 69) include quick oats, brown rice, and whole-wheat
bread. High glycemic foods (70 to 100) increase the risk of type 2 diabetes, heart
disease, obesity, and ovulatory infertility. These foods include white bread, cornflakes,
white potatoes, pretzels, rice cakes, and popcorn4.

2.1.1. Functions of Monosaccharides

In both people and plants, glucose (C 6H12O6) serves as a significant source of energy.
Carbon dioxide and water are utilized by plants to create glucose, which is then used to
meet their energy needs. Humans and herbivores ingest the starch that they store as a
result of the extra glucose. Lactose, the sugar found in milk, contains galactose, and
fructose, found in fruits and honey, gives these meals their sweetness. Ribose is a
4
Holesh, J. E., Aslam, S., & Martin, A. (2023). Physiology, Carbohydrates. In StatPearls. StatPearls Publishing.

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structural component of several coenzymes and nucleic acids. Mannose is a
component of mucoproteins and glycoproteins necessary for healthy bodily operation5.
The small intestine immediately absorbs monosaccharides into the circulation, where
they are delivered to the cells that require them. The digestive system also includes a
number of hormones, such as glucagon and insulin. By eliminating or reintroducing
glucose to the bloodstream as necessary, they keep our blood sugar levels stable.

Glycogen, a polymer similar to starch that is produced when glucose is not directly
utilised by the body, is stored in the muscles and liver as a quickly available energy
source. Our body turns glycogen back into glucose when it needs to, such as in
between meals, at night, during bursts of physical activity, or during brief fasting
intervals, to keep blood sugar levels steady.

Although the brain and red blood cells may use alternative kinds of energy from lipids
under severe conditions, such as prolonged periods of hunger, they are particularly
dependent on glucose as an energy source. Our blood glucose must be continuously
kept at an ideal level for this reason. The human brain alone requires around 130 g of
glucose each day to meet its energy requirements6.

2.1.2. Functions of Disaccharides

Disaccharides, which are starches that are present in many foods, are frequently
referred to as sugars. For instance, sucrose, or table sugar, is the most well-known
monosaccharide that humans consume. It can also be present in other foods, such as
beetroot. When monosaccharides are digested, such as sucrose, they are divided into
their direct sugars and utilized as an energy source. Bosom milk contains lactose,
5
BeMiller, J. N. (2019). Monosaccharides. Carbohydrate Chemistry for Food Scientists, 1–23.
doi:10.1016/b978-0-12-812069-9.00001-7.
6
Kumar, Pranav & Mina, Usha. (2016). Life Sciences, Fundamentals, and Practice, Part I. Retrieved from
https://www.researchgate.net/publication/335339508_Life_Sciences_Fundamentals_and_Practice_Vol_1_Seven
th_edition

10
which provides newborns with nutrition. Maltose is a sugar that is frequently included
in chocolate and other sweets.
Disaccharides like sucrose are utilized by plants to store energy, and the phloem also
transports nutrients. Numerous plants, like sugar sticks, are rich in sucrose because it is
a source of energy storage. Some parasites and green growth use trehalose for
transport. Polysaccharides, which are made up of several monosaccharides, are another
way that plants store energy6.

The most well-known polysaccharide utilized by plants for capacity is starch, which is
then broken down into maltose. The movement of disaccharides like glucose, fructose,
and galactose between cells in plants is also facilitated by monosaccharides. When
monosaccharides are combined with disaccharides, the particles are less likely to split
while being transported.

A byproduct of photosynthesis, which serves as a significant source of carbon and

energy for plants, is sucrose. Animals mostly obtain their energy from lactose. In the
digestion of starch and glycogen, maltose plays a significant intermediary role. For
insects, trehalose is a vital source of energy. The metabolism of carbohydrates requires
cellobiose. Gentiobiose is a component of certain polysaccharides and plant
glycosides5.

2.1.3 Functions of Oligosaccharides

Glycoproteins are proteins that have carbohydrates connected to them. They play a key
role in crucial processes such antigenicity, solubility, and protease resistance. As cell-
surface receptors, cell-adhesion molecules, immunoglobulins, and tumor antigens,
glycoproteins are important. Glycolipids, which are lipids with carbohydrates linked to
them, are crucial for cell identification and control membrane proteins that serve as
receptors. Cell adhesion is mediated by particular carbohydrate-binding proteins called

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lectins, which bind to oligosaccharides. In plant tissues, oligosaccharides are a part of
the fiber5.
Either glycolipids or glycoproteins cover cells. These two aid in identifying the cell
type. The lectins or proteins that join the carbs together can determine the composition
of oligosaccharides. These lectins or proteins offer some information that can be
utilized to identify cells.

The determination of blood type is an illustration of an oligosaccharide that functions

as a cell recognition agent. The glycan on the blood cell surface is modified to
distinguish all different types of blood. Mass spectrometry may be used to identify
different blood types. In mass spectrometry, antigens come in three different
categories: A, B, and H. An O blood type is indicated by the H antigen. All of the
oligosaccharides identified on all three antigens (A, B, and H) occur on the
oligosaccharide's non-reducing side.

Lectins are carbohydrate-binding proteins produced by certain cells. These lectins use
oligosaccharides to mediate cell adhesion. Cell to cell adhesion is mediated by
selectins, a class of lectins. Endothelial cells within the immune system have the
ability to express certain genes in response to cell injury or damage6.

2.1.4 Functions of Polysaccharides

Energy is stored in organisms by means of a variety of polysaccharides. While
polysaccharides often fold together and can contain several monosaccharides in a
small space, the enzymes that create energy only interact with the monosaccharides
contained in a polysaccharide. Additionally, because the side chains of the
monosaccharides create as many hydrogen bonds with one another as they can, the
molecules are hydrophobic because water cannot enter them. The molecules may hold
together and resist dissolving into the cytosol thanks to this characteristic. More sugar

12
may then be taken in since the concentration of sugar inside a cell is reduced. In
addition to storing energy, polysaccharides also enable variations in the concentration
gradient, which can affect how nutrients and water are taken up by cells5.
When they form a covalent link with proteins or lipids, many polysaccharides change
into glycoconjugates. It is possible to communicate between and inside cells using
glycolipids and glycoproteins. Certain polysaccharides may act as "tags" on proteins
that are destined for a particular organelle, assisting the cell in moving the protein
there. Special proteins that assist attach the protein, vesicle, or other molecule to a
microtubule can recognize the polysaccharides. Once tagged by certain
polysaccharides, the system of microtubules and accompanying proteins in cells may
transport any molecule to its intended place. Additionally, the immune systems of
multicellular organisms are activated by the identification of glycoproteins on cell
surfaces. A single organism's cells will create particular polysaccharides to decorate
them. The immune system is activated and kills the invader cells when it detects more
polysaccharides and distinct glycoproteins.

Support is by far one of polysaccharides' most important functions. The carbohydrate

cellulose provides support to all plants on Earth. Chitin is used by other creatures, such
as fungus and insects, to sustain the extracellular matrix surrounding their cells. To
make tissues that are more stiff, less rigid, or even materials with unique qualities, a
polysaccharide can be combined with a wide range of additional ingredients. Hundreds
of billions of tons of chitin and cellulose, both polysaccharides comprised of glucose
monosaccharides, are produced annually by living things.

Polysaccharides are the building blocks for everything, from the wood in trees to the
shells of aquatic animals. Polysaccharides can transform from storage molecules to
more stronger fibrous molecules by merely rearrangement of the structure. Most
monosaccharides' ring structures, as can be seen below, facilitate this activity.

13
Polysaccharides with a structural function give cells, organs, and organisms their
mechanical stability. Chitin and cellulose are a couple of examples. While cellulose
plays a significant role in ruminant diets, chitin is involved in the formation of fungal
cell walls.

Storage polysaccharides: These are carbohydrate reserves that release sugar monomers
when the body requires them. Glycogen, starch, and inulin are among examples. For
plants, starch serves as a kind of energy storage, while for mammals, the enzyme
amylase, which is present in saliva, catalyzes the conversion of starch into energy.
Inulin is a storage reserve in plants, whereas glycogen is a polysaccharide food reserve
of mammals, microbes, and fungus7.

Marine algae's cell walls have a supporting framework made of agarose.

The peptididoglycan found in bacterial cell walls is crucial. It contributes to binary

fission during bacterial reproduction and strengthens the cell wall. By reducing the
osmotic pressure of the cytoplasm, peptididoglycan prevents bacterial cells from
bursting. In the eye's vitreous humor and the synovial fluid that lubricates the body's
joints, hyaluronic acids play a crucial role. Additionally, it has a role in the
angiogenesis, tumor metastasis, and blood clotting processes. Heparin functions as a
natural anticoagulant to stop blood clots. Keratan sulfate is present in the cornea,
cartilage, and bones. In joints, it acts as a cushion to absorb mechanical stresses.
Chondroitin is a vital component that helps cartilage withstand compression. Dermatan
sulfate has an impact on cardiovascular problems, wound healing, blood coagulation
management, infection responses, and wound healing7.

7
European Commission JRC Knowledge Gateway. (2019). Health promotion and disease prevention. Retrived
from https://knowledge4policy.ec.europa.eu/health-promotion-knowledge-gateway_en

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2.2. Quantity
Carbohydrates are increasingly being criticized for their involvement in the epidemic
of obesity and its subsequent cardiometabolic consequences, including diabetes and
cardiovascular disease, as the worry over dietary fat has started to fade. The amount of
carbs in the diet has received a lot of attention, and conventional sources of
carbohydrates including cereal grains, pulses, and pasta have come under fire in the
popular press, best-selling books, and social media. The rise in popularity of low-carb
diets and its variations, such as the ketogenic diet, has corresponded with this negative
advertising. The relevance of carbohydrate quality is argued for in this review by
examining the data on the impact of carbohydrate quantity vs. quality in
cardiometabolic health.

The "carbohydrate-insulin model" has been put up to explain how carbohydrates may
cause obesity and its subsequent cardiometabolic problems. This model's premise is
that an excess of carbohydrates or a high ratio of carbohydrates to fat or protein causes
endocrine dysregulation characterized by hyperinsulinemia, which drives fuel
partitioning, with carbohydrates directed from metabolically active tissue (the heart,
lung, liver, etc.) to adipose tissue, resulting in a state of "cellular internal starvation"
with adaptive increases in intake and decreases in energy expenditure leading to
weight gain8.

Low-carbohydrate diets do, in fact, produce the metabolic and endocrine responses
that the model predicts they will (decreases in insulin and increases in fat oxidation,
respectively) but they do not result in the weight loss that the model predicts. When a
low-carbohydrate diet or low-carbohydrate ketogenic diet was compared to a high-

8
Ludwig, D. S., & Friedman, M. I. (2014). Increasing adiposity: consequence or cause of overeating?. JAMA,
311(21), 2167–2168. https://doi.org/10.1001/jama.2014.4133

15
carbohydrate diet, a series of meticulously controlled, randomized, inpatient feeding
trials at the National Institutes of Health did not produce the predicted increases in
total energy expenditure and body fat loss. There was instability in the effect, despite
the fact that a carefully designed longer-term randomized trial did demonstrate the
expected rise in total energy expenditure with a low-carbohydrate diet compared to a
high-carbohydrate diet during a weight loss maintenance phase after a period of weight
loss.

The effect vanished when the comparisons were made using a predetermined analysis
plan in which the baseline pre-weight loss anchor was used instead of the immediate
post-weight loss anchor (the conditions under which the doubly labelled water
measurement was validated and for which the statistical power was calculated). The
most important clinical sign, body fat, has not decreased as anticipated in any of the
clinical studies conducted to yet. It can be argued that the primary mechanism by
which low-carbohydrate diets induce weight loss is not through an increase in energy
expenditure but rather through changes in food intake regulation that result in reduced
energy intake and/or spontaneous increases in physical activity over the long term,
neither of which were assessed in these meticulously controlled clinical studies. While
these data collectively have been taken as evidence of experimental falsification of the
model, it can be argued that the principal mechanism by which low-carbohydrate diets
induce weight loss is not.

The ability to assess whether low-carbohydrate diets can cause weight reduction
through the aforementioned and/or other processes in the "real world" has been made
possible by a sizable database of long-term randomized controlled trials of dietary
guidance done under free-living settings. These studies have not demonstrated a
significant benefit of a low-carbohydrate diet over a high-carbohydrate diet. There
were no changes in weight reduction at follow-up at 6 and 12 months, according to a

16
network meta-analysis of 48 randomized trials comprising 7286 people and diets with
different distributions of macronutrients. The results of a subsequent sizable
randomized trial, the DIETFITS (Diet Intervention Examining the Factors Interacting
With Treatment Success) trial, which included 609 overweight or obese participants,
were supported by the finding that there were no differences between a "healthy" low-
carbohydrate diet and a "healthy" high-carbohydrate diet over 12 months9.

Cardiometabolic risk factors are not any better managed by low-carbohydrate diets
than by high-carbohydrate diets. The early gains in glycemic control shown at 6
months were not found to be sustained at 12 months in patients with diabetes in
systematic reviews and meta-analyses of randomized trials. Increases in the more
established and atherogenic lipid targets for lowering cardiovascular risk, low-density
lipoprotein cholesterol (LDL-C), non-high-density lipoprotein cholesterol (non-HDL-
C), and apolipoprotein B (apo B), in people with and without diabetes, have also been
shown to come at the expense of improvements seen in triglycerides and high-density
lipoprotein cholesterol.

The "Eco-Atkins" randomized trial found that a low-carbohydrate diet containing

higher quality unsaturated fat from nuts and canola oil and plant protein reduced low-
density lipoprotein cholesterol compared to a high-carbohydrate diet in 47 overweight
hyperlipidemic participants over 4 weeks while foods were provided. The quality of
the protein and fat substituting for the carbohydrate in low-carbohydrate diets,
however, is an important consideration. The most crucial factor of success in the
known randomized studies over the long term has consistently been adherence to any

9
Hall K. D. (2017). A review of the carbohydrate-insulin model of obesity. European journal of clinical
nutrition, 71(3), 323–326. https://doi.org/10.1038/ejcn.2016.260

17
one diet and clinic attendance regardless of the macronutrient distribution, regardless
of the carbohydrate content and result10.

There has been reported damage at the extremes of consumption, according to data
from extensive prospective cohort studies that allow one to evaluate the link between
carbohydrate intakes and subsequent clinical outcomes of cardiometabolic disorders of
public health and clinical concern. There was a U-shaped relationship between
carbohydrate and mortality, with low-carbohydrate (40% energy) and high-
carbohydrate (>70% energy) diets associated with increased mortality and the wide
range in between (40%-70% energy) associated with lower mortality, according to a
systematic review and meta-analysis of 5 prospective cohort studies involving 432 179
participants over a median follow-up of 25 years.

Only high-carbohydrate diets (>70% energy) were linked to an increased risk of

cardiovascular and all-cause mortality over a 10-year follow-up period in the
Prospective Urban and Rural Epidemiological Cohort Study, which included 135 335
participants free of cardiovascular disease from 18 low-, middle-, and high-income
countries. Again, a key factor was the caliber of the macronutrients used to replace the
carbohydrate. While replacing carbohydrates with animal fat or protein was linked to
an increase in mortality, replacing carbohydrates with plant-based unsaturated fats and
protein was linked to a decrease in mortality11.

It was also discovered that the carbohydrate's source was significant. According to a
concurrent publication of the Prospective Urban and Rural Epidemiological study, the

10
Ludwig, D. S., & Ebbeling, C. B. (2018). The Carbohydrate-Insulin Model of Obesity: Beyond "Calories In,
Calories Out". JAMA internal medicine, 178(8), 1098–1103. https://doi.org/10.1001/jamainternmed.2018.2933
11
Ebbeling, C. B., Feldman, H. A., Klein, G. L., Wong, J. M. W., Bielak, L., Steltz, S. K., Luoto, P. K., Wolfe,
R. R., Wong, W. W., & Ludwig, D. S. (2018). Effects of a low carbohydrate diet on energy expenditure during
weight loss maintenance: randomized trial. BMJ (Clinical research ed.), 363, k4583.
https://doi.org/10.1136/bmj.k4583

18
quality of the carbohydrate may modify the association, with higher intakes of
carbohydrate from foods like fruit and legumes being linked to lower cardiovascular
mortality and all-cause mortality rather than higher rates. Overall, these findings imply
that there is a wide range of permissible carbohydrate intakes and that assessing the
link between carbohydrate consumption and cardiometabolic outcomes may need a
focus on carbohydrate quality rather than quantity12.

2.3. Quality
Numerous indicators of the quality of carbohydrates have been identified. High-quality
carbohydrate food sources (whole grains, legumes, or fruit), low glycemic index (GI)
and glycemic load (GL), high dietary fiber, and low sugars are usually considered to
be the four key indicators. The best available data from randomized controlled trials of
intermediate cardiometabolic risk factors and prospective cohort studies of clinical
cardiometabolic disease outcomes have shown that high-carbohydrate dietary patterns
that emphasize these carbohydrate quality markers show evidence of benefits for the
prevention and management of cardiometabolic diseases13.

Systematic reviews and meta-analyses of randomized controlled trials have

demonstrated that eating habits that place a focus on fruits or dietary pulses can help
people lose or maintain their weight while also improving their blood pressure, lipid
profiles, and glycemic control. However, it appears from the systematic reviews and
meta-analyses of whole grains (which included >25 studies, >2000 individuals, and up
to 16 weeks of follow-up), that the benefits are only seen in whole grain sources made

12
Johnston, B. C., Kanters, S., Bandayrel, K., Wu, P., Naji, F., Siemieniuk, R. A., Ball, G. D., Busse, J. W.,
Thorlund, K., Guyatt, G., Jansen, J. P., & Mills, E. J. (2014). Comparison of weight loss among named diet
programs in overweight and obese adults: a meta-analysis. JAMA, 312(9), 923–933.
https://doi.org/10.1001/jama.2014.10397
13
Gardner, C. D., Trepanowski, J. F., Del Gobbo, L. C., Hauser, M. E., Rigdon, J., Ioannidis, J. P. A., Desai, M.,
& King, A. C. (2018). Effect of Low-Fat vs Low-Carbohydrate Diet on 12-Month Weight Loss in Overweight
Adults and the Association With Genotype Pattern or Insulin Secretion: The DIETFITS Randomized Clinical
Trial. JAMA, 319(7), 667–679. https://doi.org/10.1001/jama.2018.0245

19
from oats and barley. Findings from prospective cohort studies of the outcomes of
clinical cardiometabolic illness has been shown to be comparable with findings from
randomized trials targeting intermediate cardiometabolic risk variables. High intakes
of whole grains (involving >15 studies in >400 000 participants with up to 25 years'
follow-up), dietary pulses (involving >8 studies in >200 000 participants with up to 29
years' follow-up), and fruit (involving >10 studies in >500 000 participants with up to
23 years' follow-up) are linked to lower incidences of cardiovascular disease and
diabetes, according to systematic reviews and meta-analyses of prospective cohort
studies14.

Reviews and meta-analyses of more than 50 randomized controlled trials involving

more than 4000 participants revealed that low GI and GL dietary patterns, as opposed
to high GI and GL dietary patterns, result in weight loss/maintenance, clinically
significant improvements in glycemic control, as measured by a decrease in glycated
hemoglobin of less than 0.5%, as well as improvements in blood lipids and blood
pressure. This is consistent with the information from prospective cohort studies that
has been collected about the association between clinical outcomes for
cardiometabolic diseases and low GI and GL diets. Low GI and GL dietary patterns
have been linked to a lower incidence of diabetes and cardiovascular disease with up
to 25 years of follow-up, according to systematic reviews and meta-analyses of more
than 20 prospective cohort studies including more than 600 000 people.

A key biological parallel with the oral prandial drug acarbose, an alpha-glucosidase
inhibitor that successfully changes the diet to a low GI/GL eating pattern, provides
further evidence of a causative association with clinical cardiometabolic illness
outcomes. With the exception of the Cardiovascular Evaluation (ACE) trial, systematic
14
Korsmo-Haugen, H. K., Brurberg, K. G., Mann, J., & Aas, A. M. (2019). Carbohydrate quantity in the dietary
management of type 2 diabetes: A systematic review and meta-analysis. Diabetes, obesity & metabolism, 21(1),
15–27. https://doi.org/10.1111/dom.13499

20
reviews and meta-analyses of clinical outcomes trials in individuals with type 2
diabetes and large individual clinical outcome trials in individuals at risk for type 2
diabetes with impaired glucose tolerance have demonstrated that acarbose results in
comparable reductions in glycated hemoglobin and, concomitantly, reductions in type
2 diabetes, hypertension, and cardiovascular events. This experiment failed to
demonstrate a decrease in cardiovascular events with a lower dose of acarbose in
Chinese individuals with impaired glucose tolerance and pre-existing coronary heart
disease, despite a reduction in type 2 diabetes15.

The majority of the data supporting public health recommendations to restrict sugars
comes from beverages with added sugar that give out too much energy. The fructose
moiety of sugars, to which damage has been linked due to its particular set of
metabolic and endocrine reactions, is the moiety of sugars in which this finding is
clearly apparent in the available randomized controlled trials. Fructose, in energy-
matched substitutions with other carbohydrates (primarily starch), does not show
harmful effects on intermediate cardiometabolic risk factors and even shows beneficial
effects (especially for fruit) for glycemic control and blood pressure, according to
systematic reviews and meta-analyses of more than 50 randomized controlled trials
involving more than 1000 participants.

Signals for damage are only present when extra energy, which in the available studies
comes almost solely from sugar-sweetened drinks, is added to diets as opposed to the
identical diets without the added fructose and excess energy. Evidence from
prospective cohort studies evaluating the results of clinical cardiometabolic illness
supports these conditional effects. Even though systematic reviews and meta-analyses
of more than 15 prospective cohort studies involving more than 400 000 participants
15
Mansoor, N., Vinknes, K. J., Veierød, M. B., & Retterstøl, K. (2016). Effects of low-carbohydrate diets v.
low-fat diets on body weight and cardiovascular risk factors: a meta-analysis of randomised controlled trials.
The British journal of nutrition, 115(3), 466–479. https://doi.org/10.1017/S0007114515004699

21
have shown a negative association between sugar-sweetened beverages and incident
obesity, diabetes, heart disease, and stroke, this negative association is significantly
reduced with adjustment for energy (hence, many models do not adjust for energy as it
is believed to be on the causal pathway between the exposure [sugars] and the outcome
[cardiometabolic diseases])16.

High viscous soluble fiber intake from oats, barley, psyllium, and konjac mannan leads
to improvements in blood lipids, including the established therapeutic targets low-
density lipoprotein cholesterol, non-high-density lipoprotein cholesterol, and
apolipoprotein B. This is demonstrated by systematic reviews and meta-analyses of
more than 100 randomized controlled trials with more than 5000 participants. For
sources of insoluble fiber, the same has not been demonstrated. Although this data
from randomized controlled trials implies that sources of viscous soluble fiber are
most reliably connected to reductions in intermediate cardiometabolic risk variables,
there is no such differentiation in the current prospective cohort studies. High total
fiber intake, regardless of source (cereals, vegetables, or fruit) or type (insoluble vs.
soluble), is associated with a lower incidence of diabetes and cardiovascular disease
over follow-up of up to 19 years, according to systematic reviews and meta-analyses
of 10 prospective cohort studies with >1 000 000 participants17.

Other significant food sources of sugars, such as grains and grain products, dairy and
dairy products, and fruit and fruit products, have also failed to demonstrate negative
connections, and in the cases of fruit, 100% fruit juice, yogurt, and morning cereals,
have even demonstrated positive effects. When considered as a whole, the data shows
16
Jenkins, D. J., Wong, J. M., Kendall, C. W., Esfahani, A., Ng, V. W., Leong, T. C., Faulkner, D. A., Vidgen,
E., Greaves, K. A., Paul, G., & Singer, W. (2009). The effect of a plant-based low-carbohydrate ("Eco-Atkins")
diet on body weight and blood lipid concentrations in hyperlipidemic subjects. Archives of internal medicine,
169(11), 1046–1054. https://doi.org/10.1001/archinternmed.2009.115
17
Seidelmann, S. B., Claggett, B., Cheng, S., Henglin, M., Shah, A., Steffen, L. M., Folsom, A. R., Rimm, E. B.,
Willett, W. C., & Solomon, S. D. (2018). Dietary carbohydrate intake and mortality: a prospective cohort study
and meta-analysis. The Lancet. Public health, 3(9), e419–e428. https://doi.org/10.1016/S2468-2667(18)30135-X

22
that any advantage of low-sugar dietary patterns may be mediated more by energy
regulation (comparator) and the food supply (matrix) than by any unique metabolic or
endocrine mechanisms ascribed to the fructose-containing sugars they include. While
many high-quality carbohydrate food sources—including fruit, 100% fruit juice,
yogurt, and breakfast cereals—that frequently contain fructose-containing sugars show
evidence of net benefit when used as an energy-equivalent replacement for refined
starches (low-quality carbohydrate food sources) in balanced weight-maintaining diets,
sugar-sweetened beverages that provide excess energy show evidence of
cardiometabolic harm18.

3. Deficiency
Low-digestible carbohydrates are an enzyme-resistant type of saccharides with unique
effects on the human digestive system. They impact the small bowel's capacity to
digest and absorb nutrients, as well as to regulate glucose and lipid metabolism and
guard against known risk factors for illness of the heart. They are primarily broken
down in the colon by anaerobic bacteria in a process known as fermentation. As a
result, there is an increase in fecal nitrogen excretion, which is utilized to stop or treat
hepatic encephalopathy medically. Trophic are low-digestibility carbohydrates to the
ileum and colon epithelia, preventing bacterial translocation. Short-chain fatty acids
are crucial fermentation byproducts that are being considered as potential novel
treatments for acute colitis19.

18
Dehghan, M., Mente, A., Zhang, X., Swaminathan, S., Li, W., Mohan, V., Iqbal, R., Kumar, R., Wentzel-
Viljoen, E., Rosengren, A., Amma, L. I., Avezum, A., Chifamba, J., Diaz, R., Khatib, R., Lear, S., Lopez-
Jaramillo, P., Liu, X., Gupta, R., Mohammadifard, N., … Prospective Urban Rural Epidemiology (PURE) study
investigators (2017). Associations of fats and carbohydrate intake with cardiovascular disease and mortality in
18 countries from five continents (PURE): a prospective cohort study. Lancet (London, England), 390(10107),
2050–2062. https://doi.org/10.1016/S0140-6736(17)32252-3
19
Scheppach, W., Luehrs, H., & Menzel, T. (2001). Beneficial health effects of low-digestible carbohydrate
consumption. British Journal of Nutrition, 85(S1), S23-S30. doi:10.1079/BJN2000259

23
Because Carbs are one of the three main components of human nutrition and the main
source of energy for the body, if there are too few Carbs in the diet, it will have many
harmful effects on health. Constipation can occur due to lack of fiber and nutrients. At
the same time, due to lack of energy, the body will have to metabolize protein and
lipids to compensate, and protein is the raw material to build cells, leading to the
formation of new affected cells.

Although eating too little Carbs is harmful to your health, consuming too much Carbs
is absolutely not beneficial. If there is too much Carbs, blood sugar levels will
increase, and excess glucose will be stored as fat, leading to diabetes and a variety of
other health problems.

Fatigue, difficulty concentrating

- Carbohydrates are one of the main sources for energy production. If you eat foods
that contain carbs, they will immediately be available to produce energy for the
body. Without carbohydrates, the body has to go through the process of converting
sugar into carbs.

- If you don't consume enough carbohydrates, you may feel tired throughout the day!
Additionally, carbohydrates help healthy brain function. Lack of carbs can lead to
lack of concentration, memory loss, etc.

You often feel cold

- A carbohydrate-deficient diet can cause hypothyroidism. Because starch plays a
role in regulating thyroid hormone activity, it provides energy in the form of heat
to cells. Therefore, frequently feeling cold is one of the symptoms of
hypothyroidism.

24
Muscles do not grow even with exercise
- People on low-carb diets often lose muscle tissue because the body uses these
tissues to produce energy instead of carbohydrates. That's why muscles won't grow
unless you provide enough carbohydrates and protein after exercise.

Bad breath
- People on a low-carb diet put their bodies in a state of ketosis, a state that occurs
when the body doesn't have enough carbohydrates to fuel the body. When your
body goes through a state of ketosis, you may notice your breath becomes more
foul-smelling, as your salivary glands are affected.

- People on a low-carb diet put their bodies in a state of ketosis, a state that occurs
when the body doesn't have enough carbohydrates to fuel the body. When your
body goes through a state of ketosis, you may notice your breath becomes more
foul-smelling, as your salivary glands are affected.

Constipation
- Carbohydrate-rich foods are also foods that are rich in fiber content such as grains,
vegetables, beans and nuts. Therefore, if you do not eat enough of these foods, you
will also lack fiber, which causes constipation.

- Consuming enough carbohydrates from plant-based foods will support the

digestive system and reduce the risk of constipation20.

4. Metabolism, Digestion and Absorption, Energy, Interaction and More

4.1. Metabolism

20
GB Health Watch. (2023). Nutrient Carbohydrate Symptoms. GB Health Watch. Retrieved from
https://www.gbhealthwatch.com/Nutrient-Carbohydrate-Symptoms.php

25
4.1.1. Warburg effect and redox metabolism
In the human body, glucose and glycogen are involved in a complex chemical process
called carbohydrate metabolism that includes aerobic oxidation and glycolysis. The
"Pasteur Effect'' refers to the repression of glycolysis in normal mammalian cells in
anaerobic conditions. The hematopoietic microenvironment is hypoxic under
physiologically normal circumstances, which is advantageous for maintaining HSC
homeostasis. Furthermore, during illness, this hypoxia could worsen. In HSCs,
leukemia cells, and leukemia stem cells (LSCs), hypoxia-induced HIF activation has
the ability to increase glycolysis and decrease oxidative metabolism. The
microenvironment's hypoxic, hyperoxidative, acidic, and low-nutrient conditions are
made worse by the leukemia cells' uncontrolled proliferation, endangering healthy
cells. However, certain cells still prefer glycolysis even in the presence of oxygen and
fully functional mitochondria, and these cells are characterized by enhanced glucose
absorption and fermentation of glucose to lactate. Otto Warburg first hypothesized this
phenomenon in the 1920s, and it is now known as the "Warburg effect". The majority
of leukemia cells transition from aerobic oxidation to glycolysis to produce energy.
Glycolysis results in the immediate exit of pyruvate, which is then transformed into
lactate by lactate dehydrogenase and absorbed by cells. Leukemia cells are able to
effectively compete with healthy cells for nutrition thanks to their unique metabolic
process21.

4.1.2. Mechanism of aerobic glycolysis preference

Most leukemia cells prefer aerobic glycolysis, which is partly brought on by oncogenic
mutations such those that activate v-SRC, H-RAS, or MYC. More significantly, the
switch from aerobic glycolysis to OXPHOS instead of glucose metabolism lowers the
creation of reactive oxygen species (ROS), which damage cancer's capacity for self-

21
Xu, B., Hu, R., Liang, Z., Chen, T., Chen, J., Hu, Y., … Li, Y. (2021). Metabolic regulation of the bone
marrow microenvironment in leukemia. Blood Reviews, 48, 100786. doi:10.1016/j.blre.2020.100786

26
renewal. The Warburg effect also lowers the concentrations of NADPH, H+, and
NAD+, all of which are detrimental to the metabolic and proliferative ability of
leukemia cells.

An essential cofactor of GSH, NADPH keeps it in a reducing state. Reduced GSH

offers antioxidation and detoxification properties and supports healthy immunological
function. Reduced GSH may therefore shield leukemia cells from the harmful effects
of oxygen free radicals. Additionally, the cellular oxidative respiratory chain is harmed
in the absence of NADPH and H+ cofactors, which makes cells resistant to apoptosis
and genomic lesions. Furthermore, drops in NAD+ may result in the inactivation of
enzymes like pyruvate dehydrogenase, succinate dehydrogenase, and the
mitochondrial tumor suppressor deacetylases Sirtuin 3 and 5, stabilizing HIF-1, the
primary catalyst for the Warburg effect, which can boost glucose transport, glycolysis,
and lactate production.

Additionally, in leukemia cells, aerobic glycolysis may produce energy up to 100

times more quickly than OXPHOS. The Warburg effect cannot give leukemia cells
enough energy because theoretically, each unit of glucose produces 6 molecular CO2,
6 molecular H2O, and 30-32 molecular ATP units through complete glucose oxidation
in mitochondria, but only 2 molecular lactate and 2 molecular ATP units are generated
through aerobic glycolysis. However, the majority of scientists think that aerobic
glycolysis has a metabolic rate that is far higher than mitochondrial respiration,
producing more ATP through the Warburg effect than mitochondrial respiration does.

However, the amount of ATP needed for leukemia cell development and proliferation
is far lower than the amount needed to keep normal cells alive, indicating that ATP is
never in short supply during leukemia cell growth. As a result, rather of increasing the
total amount of energy produced, the Warburg effect increases the rate of energy

27
generation. Nevertheless, some scientists have suggested that decreased pyruvate
kinase activity in PKM2-expressing cells enables PEP-dependent histidine
phosphorylation of PGAM1 and may work as an alternative glycolytic pathway for
cancer cells to maintain a high rate of glycolysis, which does not produce ATP but
encourages the proliferation of cancer cells. In addition, leukemia cells have higher
glycolytic flux due to the constrained cytosolic membrane space between respiratory
chain proteins and nutrient transporters, which enables them to better utilize fructose
1,6-bisphosphate and the PTS, which includes unphosphorylated enzyme II and
histidine phosphorylatable protein, to satisfy protein needs and increase enzymatic
efficiency. However, by boosting drug efflux, repairing DNA damage, inducing drug
metabolic inactivation, altering epigenetic and drug targets, and activating survival
pathways, the Warburg effect may also help leukemia cells survive even in the
presence of medications21.

4.1.3. Acidification of the microenvironment and immune dysregulation

The microenvironment becomes more acidic as a result of increased lactate generation
brought on by increased glucose metabolism, the Warburg effect, and the increase in
hydrogen ions released by leukemia cells, which may be advantageous for leukemia
cells' invasion but detrimental to immune cells. Increased lactate levels in the
microenvironment trigger the release of vascular endothelial growth factor and
encourage neovascularization, which simultaneously enhance leukemia cells' survival
and division.

The immunological escape of leukemia has historically been facilitated by a number of

factors. First, there is a rise in immunosuppressive cells including tumor-associated
macrophages and MDSCs. Second, in order to avoid immune detection, leukemia cells
mimic normal cells by producing immune checkpoint molecules like PD-L1 or CTLA-

28
4. Third, leukemia cells reduce the expression of their surface antigens by shedding or
cleavage to avoid detection and removal by the immune system.

Through interactions between lactate and immune cells, which may also take place in
leukemia, recent advanced investigations have shown that acidity of the
microenvironment enhances the immunological escape of cancer cells. When there is
insufficient blood replenishment, tumor-infiltrating lymphocytes (TILs) and cancer
cells may compete for glucose. This might prevent T cells from proliferating and
differentiating, which would allow leukemia cells to evade the immune system and
encourage the growth and metastasis of tumors. Since glycolysis is the primary energy
metabolic pathway for T cells during growth/activation, it is well believed that a low-
glucose milieu inhibits Teff cells. Furthermore, the disparity between internal and
external lactic acid concentrations affects the ability of T cells to transfer lactate they
create. Leukemia cells emit more lactate into the milieu, which limits the excretion of
lactate produced by glycolysis from T cells. This causes the intracellular environment
of T cells to become more acidic, which prevents monocyte-macrophage development.
Additionally, elevated lactate uptake in macrophages stimulates the enzyme arginase-1
(ARGase-1), which suppresses T cell activity and development. Both DC maturation
and cytokine production, as well as the activity of cytotoxic T cells, are reduced in an
acidic bone marrow milieu. The M1 and M2 phenotypes of macrophages are
polarizable. Numerous cytokines, including interferon-, can be produced by M1
macrophages, and they can also increase the expression of costimulatory molecules
CD80 and CD86 and MHC class II cell surface molecules to better display antigens. In
addition, M1 macrophages have the capacity to enlist cytotoxic T lymphocytes to kill
leukemia cells and stop angiogenesis. On the other hand, tumor-associated
macrophages have a higher propensity to phenotypically transform into
immunosuppressive M2 macrophages, which promote the angiogenesis and spread of
leukemia. The development of tumors is favored by the maturation of antitumor M1

29
macrophages, which is inhibited by an acidic, low-pH milieu, which impacts the
transformation of T helper (Th) cells into Th2 cells. Additionally, an acidic
microenvironment destabilizes the stability of interferon made by M1 macrophages
and reduces the cytotoxicity of NK cells. Treg cells primarily rely on lipid metabolism,
therefore they are less susceptible to metabolic competition when there are scarce
resources available. This trait of Treg cells causes a relative imbalance between Teff
cells, which exert antitumor effects, and Treg cells, which suppress immune functions,
which is advantageous for the growth of hematological malignancies.

We can investigate methods to neutralize the acidic milieu in the bone marrow to
enhance immune surveillance and clearance since the low-pH bone marrow
microenvironment functions as an inhibitory environment for T cells and NK cells21.

4.1.4. Therapeutics targeting glucose metabolism

In leukemia cells, medications that block the Warburg effect or associated genes
encourage mitochondrial respiration while inhibiting aerobic glycolysis. There are
some pertinent medications being used in various hematological malignancies at the
moment.

Jitschin et al. proved that the microenvironment of chronic lymphocytic leukemia

(CLL) triggers a glycolytic shift via Notch-c-Myc signaling. It promotes leukemia cell
proliferation and treatment resistance by increasing glucose uptake and transport.
Additionally, fludarabine-induced cell death of CLL cells was enhanced by a -
secretase inhibitor that can disrupt the Notch signaling system, demonstrating a potent
anticancer effect. In BCR-ABL-positive cells, imatinib was shown to be useful
because it switched the preferred metabolic pathway from aerobic glycolysis to
oxidative respiration. Furthermore, Sun et al. demonstrated that the AMPK/STRT1
glucose metabolic pathway promotes the survival of AML cells by upregulating the

30
long noncoding RNA ANIRIL, a crucial regulator of glucose metabolism, in AML.
They also discovered that knocking down ANIRIL and AdipoR1 reduced the amounts
of glucose transporter 1 and lactate dehydrogenase A, which decreased glucose intake
and repressed aerobic glycolysis in AML cells, indicating this as a promising approach
for AML treatment. According to Poulain et al., 6-AN, a competitive inhibitor of
glucose-6-phosphate dehydrogenase, targets the pentose phosphate pathway and
inhibits mTOR complex 1 (mTORC1) signaling, which causes AML cells to take up
less glucose and perform less glycolysis. Additionally, it demonstrated a significant
anti-leukemia impact and worked in conjunction with chemotherapy. According to
Panchabhai et al., patients with multiple myeloma who have significant glycolytic
activity and a poor prognosis may benefit from therapy that simultaneously targets
aerobic glycolysis and CD147-MCT1. Ju et al. demonstrated that FLT3/ITD mutations
promote AKT-mediated mitochondrial hexokinase to boost aerobic glycolysis while
lowering mitochondrial respiration in leukemia. Additionally, in a FLT3/ITD mutant
mouse model, the combined therapy of glycolytic inhibitors that target hexokinase,
such as 3-BrOP and 2-DG, and the FLT3 inhibitor sorafenib demonstrated effective
tumor lethality21.

4.2. Absorption
4.2.1. Going to Blood Stream
To get the monosaccharides and other nutrients into the blood so they may be
delivered to the rest of the body, the small intestine cells have membranes that contain
several transport proteins. Galactose and glucose are actively transported, whereas
fructose is absorbed through assisted diffusion. The liver is the first organ to accept
glucose, fructose, and galactose. The liver absorbs them, breaks down fructose into
even smaller carbon-containing units, transforms galactose to glucose, and either
stores the glucose as glycogen or excretes it back into the blood. Hormones influence

31
how much glucose the liver exports to the blood, and you'll soon learn that even
glucose itself controls blood glucose levels22.

4.3. Energy
4.3.1. Carbohydrates as a source of energy
Cells use carbohydrates not just as structural materials, but also to transport and store
energy. Carbohydrates are classified into three types: monosaccharides (such as
glucose), oligosaccharides (such as lactose, sucrose, and maltose), and polysaccharides
(such as glycogen). The general formula of carbohydrates is Cn(H2O)n, which causes
them to appear as carbon hydrates (association of water molecules with carbon in the
ratio C1H2O1), therefore their name. The linked carbon atoms are shown to be
associated with hydrogen atoms (—H) and hydroxyl groups (—OH) when their
molecular structures are studied. Carbohydrates perform three key biochemical
functions:
- They are a source of stored energy that may be released in a form that
organisms can use.
- They are used to transfer stored energy throughout complex organisms.
- They serve as carbon skeletons that can be rearranged to produce new
molecules.

One of the most important roles of Carbs is to provide glucose. The monosaccharide
glucose, sometimes known as "blood sugar," is found in all living cells and transports
energy in humans. Cells make use of glucose as an energy source, breaking it down in
a sequence of processes that release stored energy while also producing water and
carbon dioxide; this is a cellular form of the combustion reaction. In a process known

22
American Public University. (2023). Digestion and Absorption of Carbohydrates. LibreTexts. Retrieved from
https://med.libretexts.org/Courses/American_Public_University/APUS
%3A_An_Introduction_to_Nutrition_(Byerley)/APUS%3A_An_Introduction_to_Nutrition_1st_Edition/
03%3A_Carbohydrates/3.03%3A_Digestion_and_Absorption_of_Carbohydrates

32
as cellular respiration, Glucose molecules are broken down in a sequence of reactions
by cells in order to extract energy. Not only are monosaccharides an important source
of energy for cells, but their carbon skeletons are also used as raw materials in the
synthesis of other types of tiny organic molecules, such as amino acids and fatty acids.

33
 Figure 4.3.1.1. Monosaccharides

Monosaccharides that aren't utilized right away tend to be incorporated as monomers

into disaccharides or polysaccharides. Maltose, for example, is formed by the
connecting of two molecules of glucose. Maltose, often known as malt sugar, is a beer-
brewing component. Sucrose, or table sugar, is the most common disaccharide. It is
made up of two monomers: glucose and fructose. Sucrose is commonly used by plants
to carry carbohydrates from leaves to roots and other non-photosynthetic organs.

34
 Figure 4.3.1.2. Maltose and Sucrose

Lactose, the sugar found in milk, is another disaccharide, this time a glucose molecule
connected to a galactose molecule. Disaccharides must be hydrolyzed into
monosaccharides before they can be used for energy. Polysaccharides are
macromolecules, polymers composed of hundreds to thousands of monosaccharides
linked together by glycosidic bonds. Some polysaccharides are stored and hydrolyzed
when needed for supplying monosaccharides to cells. Plants and animals both store
sugars in the form of storage polysaccharides for later use. Starch, a polymer of
glucose monomers, is stored in plants as granules within cellular structures known as
plastids. The plant may store excess glucose by synthesizing starch. The plant may
then extract glucose from this carbohydrate "bank" through hydrolysis, which breaks
down the links between its monomers. Most animals, including humans, contain
enzymes that can hydrolyze plant starch and make glucose accessible to cells as a
nutrient. The main sources of starch in the human diet include potato tubers and grains,
wheat fruits, maize (corn), rice, and various grasses. Glycogen, like starch, functions

35
as an energy storage molecule for animals. Glycogen is mostly stored in the liver and
muscle cells of vertebrates. When the demand for energy rises, glycogen and starch are
easily digested into glucose monomers, which may then be broken down to release
their stored energy. However, this stored fuel is unable to sustain an animal for long.
Glycogen stores in humans, for example, decrease in roughly a day unless
supplemented by eating. This is a problem with low-carbohydrate diets, which can
cause weakness and fatigue.

But, if glucose is required for fuel, why store it in the form of glycogen? Because
1,000 glucose molecules exert 1,000 times the osmotic pressure of a single glycogen
molecule, water enters the cells. Many organisms would invest a lot of energy
removing excess water from their cells if polysaccharides did not exist23.

23
David M. Hillis, H. Craig Heller, Sally D. Hacker, David W. Hall, Marta J. Laskowski, David E. Sadava
(2020). Life: The Science of Biology (12th edition), W. H. Freeman, 50-52, ISBN: 1319298222, 9781319298227.

36
 Figure 4.3.1.3. Structure of cellulose, starch and glycogen

4.3.2. Hydrolysis
Hydrolysis is the breakage of water (from the Greek hydro, water, and lysis, break).
The addition of a water molecule breaks the bond between the monomers. Polymers
are broken down into their component monomers during hydrolysis reactions. Water
reacts with the covalent bonds which create the polymers. A water molecule splits into
two ions (H+ and OH-) for each broken covalent bond, with the hydrogen connecting to
one monomer and the hydroxyl group attaching to the other. Within living tissues,
linkages between monomers can thus be formed and broken. The process of digestion

37
is an example of hydrolysis within our bodies. The majority of organic material in our
food is in the form of polymers, which are far too large to enter our cells. Various
enzymes react with the polymers in the digestive tract, speeding up hydrolysis.
Monomers are then released into the bloodstream and distributed to all body cells.
Dehydration reactions then can be used by these cells to combine the monomers into
new, different polymers capable of performing specific cell functions. Dehydration
and hydrolysis can also be involved in the formation and breakdown of non-polymer
molecules, such as some lipids23.

Figure 4.3.2. Hydrolysis

4.3.3. Cellular respiration

Through a series of complex processes known as cellular respiration, glucose in the
blood is taken up by your body's cells and used to produce a fuel molecule called
adenosine triphosphate (ATP). ATP can then be used by cells to power a variety of
metabolic tasks.

38
 Figure 4.3.3.1. Cellular respiration

Oxidation is used to extract energy from food, resulting in the end products carbon
dioxide and water. This process involves cellular respiration and is divided into four
stages:
- In stage 1, in the gastrointestinal (GI) tract, metabolic fuels are hydrolyzed to a
diverse set of monomeric building blocks, which include glucose, amino acids, and
fatty acids, and absorbed.
- The process of cellular respiration begins in stage 2. Various pathways in tissues
degrade the building blocks to acetyl-CoA, a common metabolic intermediate.
Most of the energy in metabolic fuels is conserved in acetyl-CoA chemical bonds
(electrons). In reducing nicotinamide adenine dinucleotide (NAD) to NADH or
flavin adenine dinucleotide (FAD) to FADH2, a smaller portion is conserved.
Reduction is the addition of electrons that could be free, part of a hydrogen atom
(H), or part of a hydride ion (H-). In the case of glucose, Glycolysis, which occurs

39
 in the cytosol, starts the degradation process by breaking glucose into two
molecules of pyruvate. The pyruvate is then oxidized to form acetyl CoA, which
enters the citric acid cycle.
- The citric acid (Krebs, or tricarboxylic acid [TCA]) cycle oxidizes acetyl-CoA to
CO2 in stage 3. The energy released during this process is mostly conserved by
converting NAD to NADH or FAD to FADH2.
- The final stage is oxidative phosphorylation. The electron transport chain (ETC)
releases the energy of NADH and FADH2 and uses it to produce ATP by an ATP
synthase. This procedure requires the use of O224.

Figure 4.3.3.2. 4 stages of extracting energy from food

4.3.4. ATP: An Important Source of Energy for Cellular Processes

24
Lisa A. Urry, Michael L. Cain, Steven A. Wasserman, Peter V. Minorsky, Rebecca Orr (2020). Campbell
Biology (12th edition), Pearson, 68-72, ISBN-13: 9780135856215

40
Adenosine triphosphate (ATP) is the cell's energy currency and is essential for
metabolism. ATP is made up of an organic molecule known as adenosine which is
linked to a string of three phosphate groups:

Figure 4.3.4.1. Structure of ATP

More specifically,ATP is composed of the sugar ribose, the nitrogenous base adenine,
and a chain of three phosphate groups (the triphosphate group) bonded to it.
Hydrolysis can break the bonds between the phosphate groups of ATP. When the
terminal phosphate bond is broken by the addition of a water molecule, a molecule of
2−¿¿
inorganic phosphate HOPO 3 (symbol: ) and an organic molecule (symbol: P) are
released. Adenosine diphosphate, or ADP, is formed when ATP loses one phosphate.
The reaction is exergonic and releases 7.3 kcal of energy per mole of ATP hydrolyzed:

G = -7.3 kcal/mol (- 30.5 kJ/mol)

This is the change in free energy measured under standard conditions. Conditions in
the cell do not meet standard criteria. When ATP hydrolysis occurs under normal
cellular conditions, the actual G is approximately -13 kcal/mol, which is 78% greater
than that of ATP hydrolysis under normal conditions.
Because the ATP releases somewhat greater energy on losing a phosphate group than
most other molecules could deliver, so it is useful to the cell. Although ATP is

41
sometimes described as storing energy, it is more accurately described as holding the
potential to react with water or other molecules. Overall, the process generates energy
that the cell can use.

Figure 4.3.4.2. The ATP cycle

The cell can use the high free energy of ATP to drive chemical reactions that are
endergonic on their own with the help of specific enzymes. If the DG of an endergonic
reaction is less than the amount of energy released by ATP hydrolysis, the two
reactions can be coupled, resulting in exergonic coupled reactions. This is usually
accomplished through phosphorylation, which is the transfer of a phosphate group
from ATP to another molecule, such as the reactant. A phosphorylated intermediate is
a recipient molecule that has a phosphate group covalently bonded to it. The key to
coupling exergonic and endergonic reactions is its formula, which is more reactive
than the original unphosphorylated molecule.

42
 Figure 4.3.4.3. How ATP drives chemical work: energy coupling using ATP
hydrolysis.

Transport and mechanical work within the cell are almost always powered by ATP
hydrolysis. ATP hydrolysis alters a protein's shape and, in some cases, its ability to
bind another molecule. Phosphorylation and dephosphorylation both promote
important protein shape changes during a variety of other critical cellular processes25.

Figure 4.3.4.4. How ATP drives transport and mechanical work

4.4. Interaction: Carbohydrate-protein interactions

Carbohydrates, which are also known as sugars or glycans, are the main source of
energy for creatures that live, serve as structural elements, and are required by
cofactors (such as ATP, NADPH, and others), glycoproteins, and polynucleotides.
These carbohydrates interact with various types of proteins and serve as the foundation
for numerous biological processes, both normal and pathological, such as fertilization,
immune response, cancer, and so on. These interactions are also important in several
cell adhesion phenomena, including the attachment of parasites, fungi, bacteria, and
viruses to host cells, which is a necessary step in infection. In many cases, the protein-
carbohydrate interaction is only the first step in a chain of events that can lead to
complex signaling cascades. Carbohydrates can play a wide range of structurally
diverse moieties from a few monosaccharides linked by various linkage types,

25
Turco, S. (2019). USMLE® STEP 1: BIOCHEMISTRY AND MEDICAL GENETICS-Lecture notes. Kaplan
Medical, a division of Kaplan Inc.

43
allowing them to play a wide range of roles in molecular recognition. Furthermore,
these carbohydrates can be extremely branched, allowing oligosaccharides to offer a
nearly infinite variety of structural variations.

Protein-carbohydrate interactions have long been recognized for their importance in

regulating a variety of physiological processes. However, the molecular basis for such
interactions has only recently been discovered. The main technique used is X-ray
crystallography for investigating protein-carbohydrate interactions, but carbohydrates
are difficult to crystallize due to their inherent flexibility, which may not be compliant
with structural analysis. They may even prevent crystallographic studies if the protein
surface's flexibility interferes with the arrangement of crystal contacts, which normally
results in their enzymatic elimination as part of the sample preparation for
crystallization. As a result, the application of NMR spectroscopy was used to obtain
conformational and dynamical information. Because of the typical characteristics of
carbohydrates, it has been established that relaxation NMR parameters should be
integrated with computational methods in order to explicitly define the structural
characteristics of carbohydrates. To better understand the structure/activity
relationships of protein-carbohydrate interactions, interdisciplinary approaches are
likely the best options for studying these complexes26.

5. Digestion
Understanding the impact of ingested food components on the human digestive system
is an area of interest to researchers because of its connection to nutrition and health.
Foods contain ingredients that may have beneficial or harmful effects on human

26
Malik, A., Baig, M. H., & Manavalan, B. (2019). Protein-Carbohydrate Interactions. In Encyclopedia of
Bioinformatics and Computational Biology (pp. 666-677). Academic Press. ISBN 9780128114322.
https://doi.org/10.1016/B978-0-12-809633-8.20661-4.

44
health. The digestive system must physically and chemically break down eaten food to
release its components, which are further metabolized for use by the body27.

Digestion is the breakdown of food into particles small enough to pass through the
cellular barrier of the gastrointestinal (GI) system and be transported throughout the
body during circulation. This occurs because of both mechanical and chemical
processes that begin in the mouth and typically end in the small intestine, where 90%
of absorption occurs. The remaining 10% takes place in the stomach and large
intestine and often with the help of intestinal microflora.

Mechanical digestion begins in the mouth with chewing and continues with partial
muscle contractions in the stomach and intestines. Chemical digestion is mainly
carried out through enzymes found in the secretions of the salivary glands, stomach,
pancreas and on the epithelial layer of the small intestine28.

5.1. Enzyme action: Digestive enzymes

Digestive enzymes, along with stomach acid, play an important role in the early stages
of digestion, i.e. the breakdown of the food we eat. The main types of human digestive
enzymes include proteases, lipases and carbohydrates, which in turn break down
macronutrients proteins, fats and carbohydrates. If we don't digest these foods
effectively, important nutrients such as essential fats, vitamins, minerals and
phytonutrients cannot be absorbed. Furthermore, Undigested or partially digested food
enters the large intestine and is fermented by resident colon bacteria, causing
unpleasant symptoms such as bloating and gas and contributes to a toxic gut29.

27
McQuilken, S. A. (2021). Digestion and absorption. Anaesthesia & Intensive Care Medicine, 22(5), 336-338.
https://doi.org/10.1016/j.mpaic.2020.12.009
28
Sensoy, I. (2021). A review on the food digestion in the digestive tract and the used in vitro models. Current
Research in Food Science, 4, 308–319. doi:10.1016/j.crfs.2021.04.004
29
Thibodeau & Patton (2007). Anatomy and Physiology, 6th Edition. ISBN: 978-0723434481

45
Digestive enzymes catalyze or activate the process that breaks down food in the mouth
and intestines. Nutrients are released and can be absorbed through the intestinal barrier
into the bloodstream. So, one of the main functions of digestive enzymes is increased
bioavailability of nutrients.

Most digestive enzymes are considered hydrolysis. They break the natural chemical
bonds in foods by the addition of water (“hydro” means water and “lysis” means
separation). They do this by binding foods (i.e. the substrate) to their active site deep
in their protein structure so the reaction can occur, breaking down the food material.
One can think of the active site of an enzyme as a pocket, just a pocket dedicated to
one type of substrate. This means that digestive enzymes have a high degree of
specificity to the type of food they act on. Enzymes can also perform more functions
than one reaction. Food is digested and released from the pocket enzyme after the
reaction ends and the process begins again.

Figure 5.1. Enzyme lock and key model30

6. Transformation of nutrients during processing and preservation

30
Avi Varma, Avi Varma. (2022). Enzymes: How they work and what they do. Medical News Today. Retrieved
from https://www.medicalnewstoday.com/articles/319704

46
6.1. Browning reaction
In foods that are thermally processed, the Maillard reaction, or nonenzymatic
browning, between carbonyl and amino groups frequently occurs. The Maillard
reaction is a desired side effect of numerous commercial and home procedures, and it
gives some cooked foods their enticing flavor and brown color. The Maillard reaction
has been known to cause the degradation of various important amino acids, including
lysine31.

6.1.1. Enzymatic browning

The spontaneous occurrence of enzyme-induced browning is common in many fruits
and vegetables. When fruits and vegetables are bruised, cut, peeled, infected, or
subjected to any abnormal circumstances, brown melanins are produced by the
oxidation of phenolic chemicals, which causes the fruits and vegetables to darken
quickly when exposed to air. Enzymatic browning is caused by polyphenol oxidase
(PPO), which is found in most fruits, vegetables, and some seafood. Along with PPO,
the related oxidative enzyme peroxidase such as catechol oxidase, and other enzymes
that create melanins and benzoquinone from natural phenols. may also cause fruit and
vegetable enzymatic browning. Polyphenol oxidase first converts phenols into
quinones, which are very susceptible to nucleophilic attacks from other proteins due to
their strong electrophilic state. Following a series of events, these quinones are
polymerized, which finally causes the development of brown pigments (melanosis) on
the food's surface. The quantity of active polyphenol oxidases in the food is a good
indicator of the rate of enzymatic browning. As a result, the majority of research into
ways to stop enzymatic browning has focused on reducing polyphenol oxidase
activity. However, not every food browning has undesirable outcomes32.

31
Miranda, G., Berna, A., & Mulet, A. (2019). Dried-Fruit Storage: An Analysis of Package Headspace
Atmosphere Changes. Foods (Basel, Switzerland), 8(2), 56. https://doi.org/10.3390/foods8020056
32
Sarmila K C (2023). Browning of Food and its Types : Maillard reaction and Caramelization. Retrieved from
https://thesciencenotes.com/browning-of-food-maillard-reaction-caramelization/

47
Figure 6.1. Example of a general reaction of polyphenols by polyphenol oxidase (PPO)
that catalyzes enzymatic browning. The production of quinones undergoes more
reactions which eventually form brown pigments on the surface of the food.

Enzymatic browning control has long been difficult for the food business. Food
enzymatic browning can be stopped or slowed down using a variety of techniques,
each of which targets a different stage of the chemical reaction. The various enzymatic
browning control methods can be divided into two main categories: physical and
chemical. Usually, several techniques are applied. Sulfites are potent anti-browning
agents, but their usage has been put on hold because of the potential risks associated
with their use. Numerous studies have been done to determine the precise forms of
regulatory mechanisms that occur when the enzymatic process is encountered.
Controlling browning also entails taking steps to restore the color of the food after it
has browned32.

6.1.1.1. Physical methods

6.1.1.1.1. Heat treatment
Due to its ability to eliminate germs and inactivate enzymes, heat treatment is the
technique that is most frequently employed to stabilize foods. The most popular
technique for inactivating vegetable enzymes is blanching. Because it results in losses
of vitamins, flavor, color, texture, carbs, and other water-soluble components, it is

48
rarely used for soft fruits. It induces denaturation and subsequently inactivation of the
enzymes but also causes the destruction of thermosensitive nutrients. By exposing the
food item to high temperatures for long enough to denature the protein, it is possible to
inactivate PPO as well as other spoilage enzymes32.

PPO's catalytic activity is generally destroyed when exposed to temperatures of 70 oC,

however the length of time needed for inactivation varies depending on the product.
Grape PPO and peroxidase both demonstrated similar thermostability, losing over 90%
of their relative activity after only 5 minutes of incubation at 78 oC and 75oC,
respectively, according to Fortea et al.'s (2009) investigation on the thermal
inactivation of these enzymes. analyzed the impacts of domestic processing and
assessed the levels of polyphenols in four cultivars of pulses popular in India. They
showed that processing decreased phenolic component concentrations by 19 - 59%. In
a study on the thermal inactivation of pineapple PPO, it was shown that after 30
minutes at 40°C, the enzyme activity decreased by about 60%. Above 75°C,
denaturation rose quickly. Therefore, residual activity was around 7% at 85C and 1.2%
at 90°C after 5 minutes. Another study examined heated apple juice from eight distinct
apple kinds at high temperature (60–90°C) and short time (20–100 seconds) (HTST)
combinations to ascertain the impact of heating conditions on enzymatic browning.
The findings demonstrated that HTST treatment at 80°C already rendered PPO
inactive, but pectinesterase activity was only lowered by half and remained inactive
even at 90°C. At 60°C, the most residual pectinesterase activity was discovered. For
heating times of 50 and 100 seconds, heating at 70°C resulted in stable pectinesterase
activity and even a little increase. Complete inactivation was seen in Victoria grape
PPO (Rapeanu et al., 2006) within 10 minutes at 70°C. After 30 minutes at 70°C, the
activity of PPO from Castanea henryi nuts was 8%32.

6.1.1.1.2. Cold treatment

49
The most popular methods for preserving food and preventing deterioration are
freezing and refrigeration. In low temperatures, browning enzyme activity, or the pace
of reaction, decreases. Thus, refrigeration aids in preserving the original appearance,
color, and flavor of fresh fruits and vegetables. Fruits and vegetables are distributed
and sold in stores while being refrigerated32.

6.1.1.1.3. Oxygen elimination

Eliminating oxygen from the surroundings slows down the browning response since
the presence of oxygen is essential for enzymatic browning. When food is preserved,
air is removed or substituted with other gasses (like N 2 or CO2), such as in modified
atmosphere packaging, wine or juice bottling, the use of impermeable films or edible
coatings, or dipping into salt or sugar solutions. This keeps the food out of direct
contact with oxygen. Plastic or other impermeable coatings keep food from
evaporating moisture and being exposed to oxygen in the air. Packaging materials
infused with antioxidants, antimicrobials, and anti-fungal agents including butylated
hydroxytoluene (BHT) and butylated hydroxyanisole (BHA), tocopherols, hinokitiol,
lysozyme, nisin, natamycin, chitosan, and -polylysine are becoming more and more
popular32.

6.1.1.1.4. Irradiation
Another way to increase food shelf life is to irradiate it using UV-C, gamma, x, and
electron beams. Ionizing radiation slows down the maturity and sprouting of produce
that will keep for a long time and kills the bacteria that cause food to spoil32.

6.1.1.2. Chemical methods

6.1.1.2.1. Acidification
Like other enzymes, browning enzymes are active within a narrow pH range. For
instance, PPO is hindered below pH = 3 and exhibits its peak activity at pH = 5-7. To

50
maintain the desired pH in food items, acidifying agents and acidity regulators are
frequently utilized as food additives. Anti-browning compounds include acidulants
such as citric acid, ascorbic acid, and glutathione. Numerous of these substances also
exhibit additional anti-browning properties, including chelating and antioxidant
activity32.

6.1.1.2.2. Antioxidants
In the food sector, antioxidants are frequently utilized as food additives. These
substances interact with oxygen and prevent the browning process from starting.
Additionally, they prevent the production of melanin and interfere with the byproducts
of the subsequent reactions. Examples of antioxidants with anti-browning
characteristics include ascorbic acid, N-acetylcysteine, L-cysteine, 4-hexylresorcinol,
erythorbic acid, cysteine hydrochloride, and glutathione32.

6.1.1.2.3. Chelating agents

Because copper is a necessary cofactor for polyphenol oxidase to operate, copper-
chelating substances prevent this enzyme from doing its job. Numerous substances
with chelating action, including citric acid, sorbic acid, polyphosphates, hinokitiol,
kojic acid, EDTA, porphyrins, polycarboxylic acids, and other proteins, have been
investigated and employed in various sectors of the food business. Some of these
substances also have additional anti-browning properties, like acidifying or antioxidant
properties. Coating materials for food packaging use hinokitiol32.

6.1.1.3. Other methods

6.1.1.3.1. Natural agents
It is recognized that a variety of natural items and their extracts, including onion,
pineapple, lemon, and white wine, might prevent or delay the browning of specific
goods. By preventing PPO activity, onion and its extract demonstrate strong anti-

51
browning characteristics. Apples and bananas have been demonstrated to benefit from
pineapple juice's anti-browning properties. Lemon juice is added to doughs to give
baked goods a brighter appearance. The anti-browning characteristics of citric and
ascorbic acids in lemon juice may help to explain this effect32.

6.1.1.3.2. Genetic modification

Arctic apples have been genetically modified to silence the expression of PPO, thereby
delaying the browning effect, and improving apple eating quality3334.

6.1.2. Non – enzymatic browning

Non enzymatic browning is a chemical process that produces a brown color in foods.
The non enzymatic browning is also caused by caramelization of sugars, interaction of
quinones with amines and amino acids32.

6.1.2.1. Caramelization
The process of caramelization, which involves browning sugar, is widely employed in
cooking because of the sweet, nutty flavor and brown color it produces. Three classes
of polymers—caramelans (C24H36O18), caramelens (C36H50O25), and caramelins
(C125H188O80)—are responsible for creating the brown hues. Diacetyl and other volatile
compounds are generated throughout the process, giving off the distinctive caramel
flavor32.

The non-enzymatic browning process of caramelization is similar to the Maillard

reaction. Caramelization is a pyrolytic reaction, not an amino acid-based one like the

33
Miranda, G., Berna, A., & Mulet, A. (2019). Dried-Fruit Storage: An Analysis of Package Headspace
Atmosphere Changes. Foods, 8(2), 56. https://doi.org/10.3390/foods8020056. PMID: 30720722; PMCID:
PMC6406843.
34
Sarmila K C (2023). Browning of Food and its Types : Maillard reaction and Caramelization. Retrieved from
https://thesciencenotes.com/browning-of-food-maillard-reaction-caramelization/

52
Maillard reaction. The monosaccharides fructose and glucose are formed during
caramelization of the disaccharide sucrose32.

Figure 6.1.2.1. Example caramelization of table sugar (sucrose) caramelizing to a

brown nutty flavor substance (furan and maltol)

6.1.2.2. The Maillard reaction

Louis Camille Maillard is credited with discovering the Maillard reaction, which is a
chemical reaction between an amino acid and a reducing sugar that frequently
necessitates the addition of heat and yields a range of aromas and scents. It develops in
food and, depending on factors such as temperature, time, humidity, pH, etc., it results
in aromas and browning, which are followed by adjustments to the organoleptic and
nutritional properties of the food. The two monosaccharides that participate most
frequently are glucose and fructose, while animal products typically contain ribose.

The Maillard reaction is a common reaction that occurs in foods that have been heated
or that have been kept at room temperature for an extended period of time. In goods
like coffee, bread, and meat where a distinctive scent and a brown color are required,
the development of the Maillard reaction is encouraged. Contrarily, the food sector has
made numerous attempts to minimize the extension of the Maillard reaction in foods

53
like milk and fruit juices, where brown colors signify a subpar product. Additionally,
the Maillard reaction is to blame for the formation of hazardous substances and the
reduction in nutritional content of many foods. Additionally, the Maillard reaction and
other widely occurring processes like lipid oxidation, caramelization, and ascorbic acid
degradation have distinct metabolic pathways. Therefore, from a technological
standpoint, fine-tuning the Maillard reaction alone is challenging. Currently, gaining
novel foods with healthier and more flavorful qualities depends on our understanding
of the shared chemical pathways of these processes35.

6.1.2.3. Biochemistry of the early maillard reaction

The initial Maillard reaction is the condensation of the free aldehyde group of
carbohydrate with either the e – amino group of lysine or hydroxylysine residues or the
α – amino group of N – terminal amino acid of protein. Only open forms of sugars
react with proteins, the carbonyl group of an acyclic monosaccharide attaching to a
protein amino group via nucleophilic attack to form a labile aldimine ( Schiff base ).
This product may hydrolyse back to glucose and protein or undergo an Amadori
rearrangement to form a 1-amino-1-deoxy fructose ( fructosamine), derivative by a
stable, though slightly reversible, ketoamine linkage (Figure 6.1.2.3). This product can
cyclise to a ring structure ( N-substituted-1-amino-dexolyte pyranose). The rate of the
Amadori rearrangement is approximately one-sixtieth that of the dissociation to
glucose and protein and also varies between proteins36.

35
Krystal, B. (2020). The Maillard reaction: What it is and why it matters. The Washington Post. Retrieved from
https://www.washingtonpost.com/news/voraciously/wp/2020/01/31/the-maillard-reaction-what-it-is-and-why-it-
matters/
36
Singh, S., Sharma, P. K., Chaturvedi, S., Kumar, P., Nannaware, A. D., Kalra, A., & Rout, P. K. (2023).
Biocatalyst for the synthesis of natural flavoring compounds as food additives: Bridging the gap for a more
sustainable industrial future. Food Chemistry, 137217. https://doi.org/10.1016/j.foodchem.2023.137217

54
 Figure 6.1.2.3. The early Maillard reaction

6.1.2.4. Biochemistry of the late Maillard reaction

The formation of the Amadori product discussed above is considered the first step in
the Maillard or browning reaction, which was first observed in 1912 when it was noted
that amino acids heated in the presence of reducing sugars develop a yellow-brown
color. The Amadori product can undergo a further series of dehydration and
rearrangement reactions. It may spontaneously fragment, releasing reactive
ketoaldehyde dicarbonyl compounds (deoxyglucosone) of which 3-deoxyglucosone is
the most important. Deoxyglucosone further reacts with amino groups of proteins to
form intra-protein cross-links known as advanced glycation end (AGE) products. AGE
products can probably also form by condensation of two Amadori products, or by
reaction of deoxyglucosone with the Amadori product. AGE products are irreversible
and accumulate over the lifetime of the protein. Most AGE products are fluorescent

55
and to date the majority of clinical data have been generated by the measurement of
protein – linked fluorescence36.

6.2. Biochemical process for food flavoring

The demand for natural tastes and aromas in food additives has steadily increased over
the past few decades. In-depth study into the use of biocatalysts for the production of
flavor and fragrance compounds has been done in response to the increased demand
for natural flavors. By employing microbial biocatalysts made from free fatty acids or
other substrates, the biotransformation process produces natural tastes and fragrances
in one direction. Consumer preference for natural flavor and fragrance manufacturing
techniques is undeniable, which directs the market's attention primarily toward the
food industry. Natural tastes are substantially more expensive than their chemically
manufactured analogues, therefore this demand cannot be met by isolating and
extracting them from natural resources. One of the common methods is the
employment of biocatalysis with the proper precursors to increase the availability of
natural flavors and smells. The technology of biocatalysis for the synthesis or
resolution of optically active intermediates was established and extensively used by
the early 2000s. Since then, biocatalysis has developed into a microbial enzyme
technology that is frequently used in enterprises for the manufacture of valuable
compounds for commerce. Great catalytic activity can be achieved by using whole
cells, cell lysates, or pure enzymes to convert a substrate into a substance known as a
bio-catalyst. However, its dependability and expense are the key barriers to its
widespread use37.

Process flavor is a food flavor created by heating different precursors together while
regulating the reaction's temperature, pH, and moisture content. The Maillard reaction

37
Orlien, V., & Bolumar, T. (2019). Biochemical and Nutritional Changes during Food Processing and Storage.
Foods (Basel, Switzerland), 8(10), 494. https://doi.org/10.3390/foods8100494

56
is the fundamental theory underlying flavor processing. This reaction is crucial for
taste creation in food processing, in addition to browning, nutritional value decrease,
toxicity, and antioxidant qualities38.

7. Control reactions occuring in food

Food is a mixture of many chemicals, including proteins, amino acids, carbohydrates,
lipids, vitamins and minerals. Since ancient times, various culinary customs have been
developed over the centuries to preserve food quality. Heat treatment, such as cooking,
promotes complex chemical reactions (non-enzymatic reactions) between food
components including the Maillard reaction, caramelization, chemical oxidation of
phenols, and maderisation39.

7.1. Maillard reaction

Among these reactions, the Maillard reaction plays an important role in the formation
of various chemicals, including toxic ones. Maillard reaction is a non-enzymatic
reaction. The Maillard reaction takes place through the interaction between
electrophilic carbonyl groups of reducing sugars and nucleophilic amino groups of
proteins/peptides/amino acids. The entire Maillard reaction process occurs in three
different stages, which are the initiation stage, propagation stage and advanced stage.
In the first stage, reducing sugars such as glucose condense with molecules containing
free amino groups (of amino acids or in proteins, mainly free amino groups of
proteins). The ε-amino group of lysine, as well as the α-amino (terminal amino acid)
groups of other amino acids yield a glycosylamine condensation product with the N-
substituted side chain rearranging to give rearranged products again Amadori (ARP)
and 1- ketone amino-1 deoxy 2 ketose. Dehydration and fragmentation of sugar
38
Flores, M., Mora, L., Reig, M., & Toldrá, F. (2019). Risk assessment of chemical substances of safety concern
generated in processed meats. Food Science & Human Wellness. https://doi.org/10.1016/j.fshw.2019.07.003
39
Krystal, Becky (2020-01-31). "The Maillard reaction: What it is and why it matters". The Washington Post.
Retrieved from https://www.washingtonpost.com/news/voraciously/wp/2020/01/31/the-maillard-reaction-what-
it-is-and-why-it-matters/

57
molecules occurs in the second step. During this step, amino acids are destroyed and 5-
hydroxymethylfurfural (HMF) fission products such as pyruvaldehyde and diacetyl are
produced. The carbonyl groups condense with the free amino groups, causing nitrogen
to incorporate into the reaction products leading to the formation of aldehydes and α-
aminoketones known as the Strecker decomposition reaction. Then, a series of
reactions take place at an advanced stage, including cyclization, dehydration,
retroaldolization, rearrangement, isomerization, and more condensation reactions,
which ultimately contribute to the appearance of of brown nitrogen polymer and co-
polymer called melanoidin (Tamanna & Mahmood, 2015). The extent of the Maillard
reaction is strongly influenced by the processing method, type of sugar, amino acids,
temperature, pH, water activity, etc39.

58
 Figure 7.1.1. Schematic representation of the three stages of the Maillard reaction
modified from Hodge and Martins et al40.

The Maillard reaction is often used to produce various foods such as bread, cakes,
meat, fish, potato products, cocoa, coffee, some dairy sweets, etc., to create attractive
color, crust texture and delicious aroma. Changing the sensory characteristics of
processed food products is highly appreciated by consumers. However, in some cases,
40
Kutzli, I., Weiss, J., & Gibis, M. (2021). Glycation of Plant Proteins Via Maillard Reaction: Reaction
Chemistry, Technofunctional Properties, and Potential Food Application. Foods, 10(2), 376. MDPI AG.
Retrieved from http://dx.doi.org/10.3390/foods10020376

59
the Maillard reaction is undesirable in products including milk and fruit juices due to
the appearance of brown color. In addition, different Maillard reaction products
(MRPs) produced at different stages of the Maillard reaction have beneficial as well as
harmful health effects depending on the type and concentration of MRP formed. Due
to its tendency to chelate metals, melanoidin has antioxidant and antibacterial
properties (Morales, Somoza, & Fogliano, 2012). However, MRPs such as HMF,
furosine, acrylamide, heterocyclic amines, etc. have been shown to have hazardous
and carcinogenic qualities (ALjahdali & Carbonero, 2019), making it important to
identify and reduce them in various products (Singh, Rao, Sharma, & Arora,
2021).The Maillard reaction also leads to an increase in reactive carbonyl compounds,
which can cause carbonyl stress and ultimately contribute to the formation of advanced
glycation end products (AGEs). Increased dietary intake of these compounds has been
linked to an increased risk of conditions such as diabetes, cancer, chronic heart failure,
Parkinson's disease and Alzheimer's disease, so measures to reduce or prevent their
growth in heat-processed foods is important. Therefore, the major challenge facing the
food industrialist is to find the best and more suitable method to produce healthy food
products with good organoleptic properties. Various strategies have been developed by
researchers including the use of polyphenols, enzymes, and non-thermal techniques to
minimize the occurrence of Maillard reactions in processed food products41.

41
Deepika Kathuria, Hamid, Sunakshi Gautam, Abhimanyu Thakur (2023). Maillard reaction in different food
products: Effect on product quality, human health and mitigation strategies, Food Control, Volume 153, 109911,
ISSN 0956-7135, Doi:10.1016/j.foodcont.2023.109911.

60
 Figure 7.1.2. Some products having Maillard reaction occurred

7.2. Caramelization
Like the Maillard reaction, caramelization is a type of browning that does not require
enzymes. Unlike the Maillard reaction, caramelization is a pyrolytic reaction, as
opposed to a reaction with amino acids. The breakdown of sugar leads to the formation
of volatile compounds (caramel smell) and brown compounds (caramel color). The
reaction can be carried out thermally and is catalyzed by acids and bases. The color
and aroma depend on the type of sugar used (i.e. mono-, oligo- or polysaccharide) and
are formed mainly through deoxyosuloses, O-heterocycle and carbocycle
intermediates as well as important sugar fragments. low molecular weight. The typical
caramel aroma is derived from cyclopentanone (cyclotene) and
hydroxymethylfuranone (furaneol)42.
42
Kroh, L. W. (1994). Caramelisation in food and beverages. Food Chemistry, 51(4), 373-379.
https://doi.org/10.1016/0308-8146(94)90188-0.

61
 Figure 7.2.1. Caramelization Reaction

Figure 7.2.2. Product having Caramelization Reaction

7.3. Chemical oxidation of phenols

Phenolic compounds are present in all fruits as a diverse group of secondary
metabolites. So, they are a component of the human diet although data on dietary
intake and metabolic fate are limited. Their role in oxidation processes, as antioxidants
or substrates in browning reactions43.

Natural phenolic compounds are abundant in plants and can be found in herbs, spices,
vegetables, fruits, wine, essential oils, olive oil, and oil seeds. Natural phenolic
compounds that have useful applications in the food industry are (i) phenolic acids
43
Kalogianni, A. I., Lazou, T., Bossis, I., & Gelasakis, A. I. (2020). Natural Phenolic Compounds for the
Control of Oxidation, Bacterial Spoilage, and Foodborne Pathogens in Meat. Foods, 9(6), 794.
doi:10.3390/foods9060794.

62
with an aromatic ring, subdivided into hydroxybenzoic acids (e.g. gallic,
protocatechuic, vanillic and syringic acids) and hydroxycinnamic acids ( e.g. p-
coumaric, caffeic and ferulic acids); the antioxidant activity of these compounds is
related to the number of hydroxyl groups in their molecules; (ii) flavonoids (e.g.,
flavanols, flavones, flavanones, flavan-3-ols, isoflavones, anthocyanins), which form
the largest subgroup of natural phenolics and whose structure consists of an arranged
15-carbon framework arranged into two phenyl rings and one heterocyclic ring; (iii)
quinones are made up of two carbonyls (e.g., benzoquinone, anthraquinone); (iv)
tannins, which are abundant in fruits and are formed by quinones or flavonoids; (v)
coumarins have a combination of benzene and alpha-pyrone rings, which are
potentially toxic but also useful for their antibacterial activity if used properly; (vi)
lignan; (vii) stilbene; and (viii) curcuminoids.

Antioxidant Capacity of Phenolic Compounds: Antioxidants prevent or hinder the

oxidation chain reaction by capturing free radicals, reducing oxygen, neutralizing
singlet oxygen (1O)2, conjugating metal ions, removing hydroperoxides and absorbing
UV rays. In contrast to synthetic phenolic antioxidants that have been used for decades
in the food industry and are accused of being toxic and carcinogenic, natural phenolic
compounds have beneficial effects on the body, human health by protecting against
chronic diseases related to oxidative stress. Table 1 presents the applications of
antioxidant phenolic compounds in meat products, the effects of phenolic compounds
on lipid and protein oxidation as well as the sensory properties of the products (color,
taste and smell).

63
Table 7.3. Antioxidant effect of phenolic compounds in meat and meat-based products

7.4. Maderisation
Maderisation is a process that involves heating and oxidizing wine. The term is named
after the process used in Madeira winemaking, where it occurs while the wine is in
barrel. The resulting wine will be darker in color and have a Sherry-like character.
Outside of Madeira wines, it is often considered a fault in the wine, but is expected in
the case of certain dessert wines during aging in the bottle.

8. Technology: Protecting Our Food

64
Food safety has become an important topic. People are now concerned about the safety
of food. Technology is acting as an effective enabler in life. Some of the technologies
have been applied to better protect food:

- Intelligence cold chain systems ensure food quality and safety through
comprehensive real-time monitoring of food status in the food supply chain.
These systems organically combine sensors with Internet of Things technology.
Sensors monitor temperature, humidity and operating status of refrigeration
equipment in real-time where food is placed. Data is transmitted to the data
processing module in real time using wireless communication technology. After
processing and analyzing the data, the data will be transmitted to the server and
client. The server can conduct early warnings and make regulations according
to the results of data analysis, and the client can conduct real-time queries on
food logistics information, food status and scheduling information food history.
To achieve cold chain system integrity, sensors and their data transmission are
indispensable44.

- Microfluidic Detectors:Botulinum bacteria produce the most toxic toxin known.

They and similar agents, such as tetanus, can be detected during food
processing using microfluidic chips-self-contained diagnostic laboratories about
the size of a finger. The University of Wisconsin–Madison is building such a
chip, lined with antibodies immobilized by magnetic particles, that could detect
botulism in milk production. The chip can sample milk before or after it is
loaded into tankers leaving the dairy facility and before or after it is pasteurized
at the manufacturing plant. Other types of chips can detect other toxins in a

44
Bai, L., Liu, M., & Sun, Y. (2023). Overview of Food Preservation and Traceability Technology in the Smart
Cold Chain System. Foods, 12(15), 2881. https://doi.org/10.3390/foods12152881.

65
 variety of liquid handling plants, such as those that produce apple juice, soup,
or baby formula.

- Active Packaging: E. coli, salmonella and other pathogens can be detected

through small windows in packaging, such as cellophane around meat or plastic
jars around peanut butter. The “smart” window will contain antibodies that bind
to enzymes or metabolites produced by microorganisms, and if that happens the
patch will turn color. The challenge is to fabricate windows from materials and
reagents that can safely come into contact with food. Similar biosensors can
react if contents reach certain pH levels or are exposed to high temperatures,
indicating the food has spoiled and they can sense whether the packaging has
been tampered with, for example, by responding to the pressure created by the
syringe or to oxygen seeping through a puncture.

- RFID Tags: Pallets or boxes containing select foods now have radio frequency
identification (RFID) tags that, when read by a scanner, indicate which farm or
processing plant the shipment came from. Future tokens that are smaller,
smarter and cheaper could adorn individual packages and record every facility
they have passed through and when. The University of Florida is inventing tags
that can be read through liquids (traditional designs cannot) and so can be
attached to the inside of sour cream or yogurt containers. The university is also
developing activity tags that can record the temperature to which a package has
been exposed.

- Edible Tags: Manufacturers often combine crops from multiple growers, such
as spinach leaves, into one retail package, so tags attached to bags may not help
investigators trace contamination to a specific source. can. ARmark
authentication technology can print microscopic markers indicating the location

66
 of origin directly onto spinach leaves, apples or dog food pellets using a spray
made from edible materials such as cellulose, oil plant or protein. In addition,
the small size makes it difficult for terrorists to counterfeit, making it difficult
for them to get fake food containing toxins past inspectors and into the supply.
Instead, DataLase can spray an edible film onto half-inch-diameter fruits or
citrus meats, then shine a laser beam to burn an identification code inside the
film.

9. Dietary Fiber: Digestion

Dietary fiber consists of non-digestible forms of carbohydrate, usually as
polysaccharides that originate from plant-based foods45. Fibers have been extensively
studied over the past few decades for their physiological health benefits. Depending on
its solubility, fiber is classified into two groups - soluble and insoluble fiber. Soluble
fiber includes β-glucan, galactomannan, pectin, psyllium, inulin and resistant starch,
while insoluble fiber includes cellulose, hemicellulose, chitosan, lignin, etc. Dietary
fiber is characterized by several physicochemical properties such as solubility,
fermentability, viscosity, water absorption capacity, binding capacity, etc. These
properties are responsible for the functional activity of fiber46.

When ingested in the diet, soluble fiber increases viscosity mixed with water and leads
to a decrease in blood sugar response and plasma cholesterol. Insoluble fibers are
characterized by their porosity and low density. Due to these properties, insoluble fiber
results increases fecal bulk and reduces intestinal transit. In the food market, more
attention is paid to the development of food products enhanced with soluble fiber is
beneficial because it provides viscosity and gelling ability compared to
45
Barber, T. M., Kabisch, S., Pfeiffer, A. F. H., & Weickert, M. O. (2020). The Health Benefits of Dietary Fibre.
Nutrients, 12(10), 3209. https://doi.org/10.3390/nu12103209.
46
Mudgil, D. (2017). Chapter 3 - The Interaction Between Insoluble and Soluble Fiber. In R. A. Samaan (Ed.),
Dietary Fiber for the Prevention of Cardiovascular Disease (pp. 35-59). Academic Press. ISBN
9780128051306. https://doi.org/10.1016/B978-0-12-805130-6.00003-3.

67
insoluble fiber46.

Figure 9.1. Some foods containing Fiber47

Much scientific evidence has accumulated on the relationship between fiber and body
weight and overall metabolic function (including its effects on glucose regulation,
lipids, and insulin sensitivity). Perhaps it stems at least in part from the known
association between dietary fiber intake and a healthy gut microbiome, favorable body
weight, and overall metabolic health. is also associated with a reduced risk of
developing cardiovascular disease and death. There have also been descriptions of
further health benefits of fiber, including reduced risk of malignancy and improved
colon health46.

47
InsideTracker. (2023). Soluble Fiber Simplified. InsideTracker Blog. Retrieved from
https://blog.insidetracker.com/soluble-fiber-simplified

68
 Table 9.1. Summary of the health benefits of dietary fiber

Table 9.2. Adequate dietary fiber intake by gender and age48

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77