Research Update:Mulberry Leaf.

seminal trace...Mulberry Leaf Extract.10:1.Morus alba L.,Folium Mori.CAS.NO:94167-05-2,90064-11-

2.Synonyms:Mulberry Extract.Extract of mulberry,Morus nigra, ext....

Phytochemical info of Mulberry Leaf.

Product Name:
Synonym:
Definition:Mulberry Leaf are majorly composed of
Chemical information disclosed as following table:

Research Update:Mulberry Leaf.

Food-Grade Mulberry Powder Enriched with 1-Deoxynojirimycin Suppresses the

Elevation of Postprandial Blood Glucose in Humans.:J Agric Food Chem. 2007 Jul
11;55(14):5869-5874. Epub 2007 Jun 8.

Mulberry 1-deoxynojirimycin (DNJ), a potent glucosidase inhibitor, has been hypothesized to be

beneficial for the suppression of abnormally high blood glucose levels and thereby prevention of
diabetes mellitus. However, DNJ contents in commercial mulberry products were as low as
about 0.1% (100 mg/100 g of dry product), implying that the bioavailability of DNJ might not be
expected. We carried out studies in two directions: (1) production of food-grade mulberry
powder containing a maximally high DNJ content; (2) determination of the optimal dose of the
DNJ-enriched powder for the suppression of the postprandial blood glucose through clinical
trials. The following method was used: (1) DNJ concentrations in mulberry leaves from different
cultivars, harvest seasons, and leaf locations were determined using hydrophilic interaction
chromatography with evaporative light scattering detection. (2) Healthy volunteers received 0,
0.4, 0.8, and 1.2 g of DNJ-enriched powder (corresponding to 0, 6, 12, and 18 mg of DNJ,
respectively), followed by 50 g of sucrose. Before and 30-180 min after the DNJ/sucrose
administration, plasma glucose and insulin were determined. The following results were
obtained: (1) Young mulberry leaves taken from the top part of the branches in summer
contained the highest amount of DNJ. After optimization of the harvesting and drying processes
for young mulberry leaves (Morus alba L. var. Shin ichinose), DNJ-enriched powder (1.5%) was
produced. (2) A human study indicated that the single oral administration of 0.8 and 1.2 g of
DNJ-enriched powder significantly suppressed the elevation of postprandial blood glucose and
secretion of insulin, revealing the physiological impact of mulberry DNJ (effective dose and
efficacy in humans). This study suggests that the newly developed DNJ-enriched powder can be
used as a dietary supplement for preventing diabetes mellitus. Keywords: 1-Deoxynojirimycin;
HILIC-ELSD; mulberry leaves; Morus spp.; diabetes prevention.

Mulberry leaf powder prevents atherosclerosis in apolipoprotein E-deficient

mice.:Biochem Biophys Res Commun. 2007 Jul 6;358(3):751-6. Epub 2007 May 7.

Mulberry is commonly used to feed silkworms. Here we examined whether a dietary intake of
mulberry leaf (ML) could affect atherogenesis in vivo and in vitro. Apolipoprotein E-deficient
mice were fed either normal chow (control group) or a diet containing 1% ML powder (ML
group) from 6 weeks of age. The mice were sacrificed after 12 weeks. The susceptibility of
plasma lipoprotein to oxidation was assessed using diene formation. A significant increase in the
lag time of lipoprotein oxidation was detected in the ML group compared with the control group.
Furthermore, the ML group showed a 40% reduction in atherosclerotic lesion size in the aortae
compared with the control. We also examined the direct anti-oxidative activity of ML in vitro.
Aqueous extract of ML had a strong scavenging effect on 1,1-diphenyl-2-picrylhydrazyl and
inhibited lipoprotein oxidation. These results confirm that ML contains anti-oxidative substances
that might help prevent atherosclerosis.

Resource recovery potential from secondary components of segregated municipal solid

wastes.:Environ Monit Assess. 2007 May 15;Chanakya HN, Ramachandra TV,
Vijayachamundeeswari M.Centre for Sustainable Technologies and Centre for Ecological
Sciences, Indian Institute of Science, Bangalore, 560 012, India, cestvr@ces.iisc.ernet.in.

Fermentable components of municipal solid wastes (MSW) such as fruit and vegetable wastes
(FVW), leaf litter, paddy straw, cane bagasse, cane trash and paper are generated in large
quantities at various pockets of the city. These form potential feedstocks for decentralized biogas
plants to be operated in the vicinity. We characterized the fermentation potential of six of the
above MSW fractions for their suitability to be converted to biogas and anaerobic compost using
the solid-state stratified bed (SSB) process in a laboratory study. FVW and leaf litter (paper
mulberry leaves) decomposed almost completely while paddy straw, sugarcane trash, sugarcane
bagasse and photocopying paper decomposed to a lower extent. In the SSB process between 50-
60% of the biological methane potential (BMP) could be realized. Observations revealed that the
SSB process needs to be adapted differently for each of the feedstocks to obtain a higher gas
recovery. Bagasse produced the largest fraction of anaerobic compost (fermentation residue) and
has the potential for reuse in many ways.

Gluconeogenic substrates and hepatic gluconeogenic enzymes in streptozotocin-diabetic

rats: effect of mulberry (Morus indica L.) leaves.:J Med Food. 2007 Mar;10(1):41-8.Andallu B,
Varadacharyulu NC.Department of Home Science, Sri Sathya Sai Institute of Higher Learning,
Anantapur, Andhra Pradesh, India. andallusss@rediffmail.com

Mulberry (Morus indica L.) leaves, the sole food of the silk worm, were evaluated for
antidiabetic effects in streptozotocin (STZ)-diabetic rats. Treatment with dried mulberry leaf
powder at 25% of the diet for a period of 8 weeks was found to be remarkably beneficial to STZ-
diabetic rats as evidenced by controlled hyperglycemia and glycosuria. In addition, mulberry
leaves countered (reversed) the alterations in gluconeogenic substrates in STZ-diabetic rats as
indicated by significant reduction in serum pyruvic and lactic acid levels, a significant increase
in proteins and a significant decrease in free amino acid, urea, and creatinine levels in blood, and
a decreased urinary excretion of urea and creatinine. Anomalies in the activities of hepatic
gluconeogenic enzymes associated with impaired glucose homeostasis in STZ-diabetic rats were
ameliorated by feeding the mulberry leaf-supplemented diet, indicating that control over
hyperglycemia and associated complications in the diabetic state by mulberry leaves is by way of
regulation of gluconeogenesis. With respect to all the parameters, mulberry leaves were more
effective than the oral hypoglycemic drug glibenclamide.

Mulberry leaf extract prevents amyloid beta-peptide fibril formation and

neurotoxicity.:Neuroreport. 2007 May 28;18(8):813-6.

Mulberry leaf has been reported to possess medicinal properties, including hypoglycemic,
hypotensive and diuretic effects. Little is known, however, about its medicinal properties for
central nervous system disorders, including Alzheimer's disease. Accumulating evidence
suggests that amyloid beta-peptide (1-42) plays an important role in the etiology of Alzheimer's
disease. Here we show that mulberry leaf extract inhibits the amyloid beta-peptide (1-42) fibril
formation by both the thioflavin T fluorescence assay and atomic force microscopy.
Furthermore, mulberry leaf extract protected hippocampal neurons against amyloid beta-peptide
(1-42)-induced cell death in a concentration-dependent manner. These results suggest that
mulberry leaf extract provides a viable treatment for Alzheimer's disease through the inhibition
of amyloid beta-peptide (1-42) fibril formation and attenuation of amyloid beta-peptide (1-42)-
induced neurotoxicity.

Hypolipidemic effect of flavonoids from mulberry leaves in triton WR-1339 induced

hyperlipidemic mice.:Asia Pac J Clin Nutr. 2007;16 Suppl 1:290-4.Chen J, Li X.Faculty of Life
Science, City College, Zhejiang University, 51 Huzhou Street, Hangzhou, Zhejiang, China
310015. lixr@zucc.edu.cn.

The aim of this study was to investigate the hypolipidemic effect of the total flavonoids from
mulberry leaves (MTF) in hyperlipidemic mice. The total flavonoids have been isolated from
mulberry (Morus alba L.) leaves and purified by macroporous resins. After that, the content of
MTF is 581.7 mg g-1 in dried product. The hypolipidemic effect of MTF has been evaluated in
triton WR-1339 (400 mg kg-1) induced hyperlipidemic mice. The beneficial effects of MTF on
serum lipid levels are more significant at 12h post MTF administration than at 6h. The levels of
TG, TC and LDL-C were remarkably reduced to 388, 257 and 189 mg 100 ml-1 in MTF (30 mg
kg-1) and triton WR-1339 treated mice, compared with 540, 464 and 299 mg 100 ml-1,
respectively, in group treated by triton WR-1339 only. The ratios of HDL-C/TC and HDL-
C/LDL-C were increased to 0.42 and 0.57 post MTF (30 mg kg-1) administration, whereas these
two ratios at the low levels of 0.33 and 0.52, respectively, in the reference group. These findings
support a serum a lipid-favourable activity for mulberry leaf flavonoids.

Pathogenicity of Fusarium semitectum against crop pests and its biosafety to non-target
organisms.:Commun Agric Appl Biol Sci. 2006;71(2 Pt B):465-73.Mikunthan G, Manjunatha
M.Dept. of Agricultural Biology, Faculty of Agriculture, University of Jaffna, Sri Lanka.
mikunthan@jfn.ac.lk

Microbial control is receiving more attention, since these alternative tactics, compared to
chemical control methods, are energy saving, non polluting, ecologically sound and sustainable.
A mycopathogen, Fusarium semitectum Berk. and Rav. (ARSEF 7233) was isolated from
diseased cadavers of aphid (Aphis gossypii) and cultured in Saboraud Maltose Agar
supplemented with Yeast extract medium (SMAY). Being isolated first time from the chilli
ecosystem its potential was evaluated. Experiments were conducted to understand its
pathogenicity against crop pests as well as to ensure its safety to non target organisms such as
silk worm (Bombyx mor), honey bee (Apis indica) and earthworm (Eisenia foetida). A paper-
thrips-paper sandwich method for thrips and detached-leaf bioassay method for mites were used.
Test insects and mites either reared in laboratory or obtained from the field were topically
applied with spore suspension of F. semitectum (1x10(9) spores/ml). Mortality was recorded and
dead animals were surface sterilized with 0.5% NaOCl and placed in SMAY medium to confirm
pathogenicity. Mulberry leaves sprayed with the fungal suspension were fed to larvae of B. mori
and reared. Newly emerged A. indica were topically applied with fungus. The fungus grown in
cow dung for two weeks was used to assess the composting ability of E. foetida. F. semitectum
produced mycosis and caused mortality to sucking pests such as chilli thrips (Scirtothrips
dorsalis), broad mite (Polyphagotarsonemus latus), sugarcane wooly aphid (Ceratavacuna
lanigera), spiraling whitefly (Aleyrodicus disperses), whitefly (Bemisia tabaci, A. gossypii and
coconut mite (Aceria guerroronis). The fungus did not cause mortality on larvae of lepidopteran
insect pests and ladybird beetle (Menochilus sexmaculatus), predatory mite (Amblysius ovalis)
and larval parasitoid (Goniozus nephantidis). F. semitectum failed to infect the larvae of B. mori
and newly emerged A. indica and its brood. The mycopathogen had no influence on the
composting ability and growth of E. foetida. F. semitectum, in general, expressed its selectivity
against sucking pests and proved its eco-friendly characteristics to the beneficial organisms and
especially safe to Sericulture, Apiculture and Vermiculture industries in Karnataka, India. This
novel fungus can be well incorporated as a viable tactics into the integrated management
programmes of crop pests.

Microwave-assisted extraction of flavonoids in mulberry leaf and root-bark.:Zhong Yao

Cai. 2006 Oct;29(10):1090-2. Chinese. Chen JJ, Li XR.College of Pharmaceutical Science,
Zhejiang University, Hangzhou 310031, China.

OBJECTIVE: To extract flavonoids in Mulberry leaf and White Mulberry root-bark by

microuave-assisted method. METHODS: An orthogonal table was used to find the best
procedure for microwave-assisted extraction (MAE) of flavonoids. The advantages of MAE were
explored by comparing with the commonly used extraction methods. The different contents of
flavonoids in Mulberry Leaf and Mulberry Root-bark were also compared. RESULTS: The best
condition for procedure was extracting 20 min at 60 degrees C with 12 times of 70% ethanol.
Among all factors, the ethanol concentration was significant This experiment implied that
flavonoids in Mulberry leaf were 9 times as that in Mulberry Root-bark probably. Moreover, the
flavonoids extracted from Mulberry Leaf and White Mulberry root-bark by MAE were 55% and
32% more than that from commonly used extraction method individually. CONCLUSION: MAE
is advantageous than commonly used extraction method. It can be used for batch production.

Enzymatic formation of unnatural cytokinin analogs by adenylate isopentenyltransferase

from mulberry.:Biochem Biophys Res Commun. 2007 Apr 13;355(3):795-800. Epub 2007 Feb
15.

A cDNA encoding adenylate isopentenyltransferase (AIPT) was cloned from young leaves of
mulberry (Morus alba) by a homology-based RT-PCR. A recombinant enzyme expressed in
Escherichia coli catalyzed prenyl transfer from DMAPP to the N6 amino group of ADP and
ATP, respectively, while AMP was a poor substrate of the enzyme. Interestingly, M. alba AIPT
also accepted dADP, dATP, CDP, and GDP as the prenyl acceptors, and IPP, HMBPP, and GPP
as the prenyl donors, to produce a series of cytokinin analogs. In particular, it was remarkable
that the enzyme accepted HMBPP to produce trans-zeatin riboside phosphates, which suggested
that trans-zeatin may be also produced from adenosine phosphates and HMBPP. Finally, alanine-
scanning mutagenesis of conserved D49, Y54, F93, F120, Y153, F157, W159, Y170, Y217, and
Q255, resulted in significant loss of enzyme activity except Y170A, confirming the functional
and structural importance of the residues.

Nutritional quality of leaves of some genotypes of mulberry (Morus alba).:Int J Food Sci
Nutr. 2006 Aug-Sep;57(5-6):305-13.Srivastava S, Kapoor R, Thathola A, Srivastava
RP.Department of Foods and Nutrition, College of Home Science, U.S. Nagar, Uttaranchal,
India. sarita.srivastava@rediffmail.com

This study was undertaken to determine the proximate composition, vitamins, minerals and the
antinutritional factor tannic acid in leaves of six genotypes of mulberry. The results showed that
in fresh mulberry leaves the proximate composition values ranged from 71.13 to 76.68% for
moisture, from 4.72 to 9.96% for crude protein, from 4.26 to 5.32% for total ash, from 8.15 to
11.32% for Neutral Detergent Fiber (NDF), from 0.64 to 1.51% for crude fat, from 8.01 to
13.42% for carbohydrate and from 69 to 86 kcal/100 g for energy. In dried mulberry leaf
powder, moisture ranged from 5.11 to 7.24%, crude protein from 15.31 to 30.91%, total ash from
14.59 to 17.24%, NDF from 27.60 to 36.66%, crude fat from 2.09 to 4.93%, carbohydrate from
9.70 to 29.64% and energy from 113 to 224 kcal/100 g. Among vitamins ascorbic acid and beta-
carotene were found to range from 160 to 280 mg/100 g and from 10,000.00 to 14,688.00
microg/100 g, respectively, in fresh mulberry leaves and from 100 to 200 mg/100 g and from
8438.00 to 13,125.00 microg/100 g, respectively, in dried mulberry leaf powder. The minerals
iron, zinc and calcium were observed in the ranges of 4.70-10.36 mg/100 g, 0.22-1.12 mg/100 g
and 380-786 mg/100 g, respectively, for fresh mulberry leaves, and 19.00-35.72 mg/100 g, 0.72-
3.65 mg/100 g and 786.66-2226.66 mg/100 g, respectively, for dried mulberry leaf powder. The
tannic acid ranged from 0.04 to 0.08% in fresh leaves and from 0.13 to 0.36% in dried leaf
powder.
 Alpha-glucosidase inhibitory effect of mulberry (Morus alba) leaves on Caco-
2.:Fitoterapia. 2006 Dec;77(7-8):568-73. Epub 2006 Sep 22.Hansawasdi C, Kawabata
J.Department of Agro-Industry, Faculty of Agriculture, Natural Resources and Environment,
Naresuan University, Phitsanulok, Thailand. cicake@yahoo.com

The effects of brewing time on dry weight content and alpha-glucosidase inhibitory active
component released from mulberry (Morus alba) tea were studied. Different tea products showed
significant differences in inhibitory activity against both sucrase and maltase. The most effective
enzyme inhibition was observed when 3 to 5 min brewing time was applied in tea preparation. In
a Caco-2 cell culture experiment the tea reduced the liberated glucose contents in both apical and
basal sides of the cell monolayers. It can be concluded that hot water extract of mulberry leaves
does have inhibitory effect against alpha-glucosidases, sucrase and maltase enzymes, and has a
potential to be consumed as antidiabetic herb tea.

Mulberry leaf aqueous fractions inhibit TNF-alpha-induced nuclear factor kappaB (NF-
kappaB) activation and lectin-like oxidized LDL receptor-1 (LOX-1) expression in vascular
endothelial cells.:Atherosclerosis. 2007 Jul;193(1):20-7. Epub 2006 Oct 20.Mikunthan G,
Manjunatha M.Dept. of Agricultural Biology, Faculty of Agriculture, University of Jaffna, Sri
Lanka. mikunthan@jfn.ac.lk

Microbial control is receiving more attention, since these alternative tactics, compared to
chemical control methods, are energy saving, non polluting, ecologically sound and sustainable.
A mycopathogen, Fusarium semitectum Berk. and Rav. (ARSEF 7233) was isolated from
diseased cadavers of aphid (Aphis gossypii) and cultured in Saboraud Maltose Agar
supplemented with Yeast extract medium (SMAY). Being isolated first time from the chilli
ecosystem its potential was evaluated. Experiments were conducted to understand its
pathogenicity against crop pests as well as to ensure its safety to non target organisms such as
silk worm (Bombyx mor), honey bee (Apis indica) and earthworm (Eisenia foetida). A paper-
thrips-paper sandwich method for thrips and detached-leaf bioassay method for mites were used.
Test insects and mites either reared in laboratory or obtained from the field were topically
applied with spore suspension of F. semitectum (1x10(9) spores/ml). Mortality was recorded and
dead animals were surface sterilized with 0.5% NaOCl and placed in SMAY medium to confirm
pathogenicity. Mulberry leaves sprayed with the fungal suspension were fed to larvae of B. mori
and reared. Newly emerged A. indica were topically applied with fungus. The fungus grown in
cow dung for two weeks was used to assess the composting ability of E. foetida. F. semitectum
produced mycosis and caused mortality to sucking pests such as chilli thrips (Scirtothrips
dorsalis), broad mite (Polyphagotarsonemus latus), sugarcane wooly aphid (Ceratavacuna
lanigera), spiraling whitefly (Aleyrodicus disperses), whitefly (Bemisia tabaci, A. gossypii and
coconut mite (Aceria guerroronis). The fungus did not cause mortality on larvae of lepidopteran
insect pests and ladybird beetle (Menochilus sexmaculatus), predatory mite (Amblysius ovalis)
and larval parasitoid (Goniozus nephantidis). F. semitectum failed to infect the larvae of B. mori
and newly emerged A. indica and its brood. The mycopathogen had no influence on the
composting ability and growth of E. foetida. F. semitectum, in general, expressed its selectivity
against sucking pests and proved its eco-friendly characteristics to the beneficial organisms and
especially safe to Sericulture, Apiculture and Vermiculture industries in Karnataka, India. This
novel fungus can be well incorporated as a viable tactics into the integrated management
programmes of crop pests.

Effect of fortification of Mulberry leaves with homeopathic drug Nux vomica on Bombyx
mori. L.:Homeopathy. 2006 Jul;95(3):148-50.Hiware CJ.Department of Zoology, Dr. Babasaheb
Ambedkar Marathwada University, Aurangabad 431004, MS, India. drhiware@rediffmail.com

Silk worm (Bombyx mori L.) larvae were fed on Mulberry leaves treated with Nux vomica
mother tincture. The impact on larval, cocoon, shell and pupal weight, silk ratio, average
filament length and denier, and number of breakages during reeling were investigated. The
results were positive in all parameters under study except cocoon weight, pupal weight, and the
average denier of the filament.

Effects of oral administration of some herbal extracts on food consumption and blood
glucose levels in normal and streptozotocin-treated diabetic rats.:Methods Find Exp Clin
Pharmacol. 2006 May;28(4):223-8.Musabayane CT, Bwititi PT, Ojewole JA.Department of
Human Physiology, Faculty of Health Sciences, University of KwaZulu-Natal, Durban, South
Africa. musabayanec@ukzn.ac.za

Previous studies in our laboratories suggest that oral administration of some herbal extracts
reduce blood glucose concentrations in rats, possibly by interfering with food consumption
and/or gastrointestinal absorption of food. Accordingly, we monitored the amounts of food
consumed and body weights in separate groups of nondiabetic and streptozotocin-treated diabetic
rats, orally treated with some plant extracts (20 mg 100 g -1 body weight) daily for 5 weeks.
Control animals were administered the vehicle, citrate buffer (0.1 ml 100 g -1 body weight).
Separate groups of rats administered allopathic hypoglycemic drugs metformin (50 mg 100 g -1
body weight) or glibenclamide (5 microg 100 g -1 body weight) acted as positive control
animals. After 5 weeks, blood glucose concentrations were reduced in all the groups.
Tapinanthus nyasicus leaf, Ficus thoningii bark, Solanum incanum fruit, and Morus alba leaf
extracts decreased weekly food consumption throughout the 5-week study period. Similar results
were obtained for the groups treated with metformin or glibenclamide. However, food
consumption was increased by S. incanum root, Aloe chabaudii leaf, or Allium sativum bulb
extracts, and this was associated with high prevalence of diarrhea. The herbal extracts and
metformin did not affect serum insulin concentration in nondiabetic rats, while glibenclamide
increased serum insulin concentration. In conclusion, it may be inferred that the herbal extracts
examined produced hypoglycemia, probably by interfering with either food intake or
gastrointestinal glucose absorption (as reported for metformin). These findings merit long-term
investigation.

Inhibitory effects of extractives from leaves of Morus alba on human and rat small
intestinal disaccharidase activity.:Br J Nutr. 2006 May;95(5):933-8.

The inhibitory effect on human and rat intestinal disaccharidase by the extractive from the
leaves of Morus alba (ELM) containing 0.24 % 1-deoxynojirimycin equivalent and its inhibitory
activities were investigated by the modified Dahlqvist method. In the presence of 1000-fold
diluted ELM solution, the sucrase activity of four human samples was inhibited by 96 % and that
of maltase and isomaltase by 95 and 99 %, respectively. The activities of trehalase and lactase
were inhibited by 44 and 38 %, respectively. The human disaccharidase activities varied from
sample to sample because the samples were obtained from different resected regions after
surgery. However, the ratio of the inhibitory effect for sucrase, maltase, isomaltase, trehalase and
lactase was very similar among the four samples, and also that of resembled rat intestinal
disaccharides. The inhibitory constant of the 1-deoxynojirimycin equivalent for sucrase, maltase
and isomaltase was 2.1 x 10(-4), 2.5 x 10(-4) and 4.5 x 10(-4) mm, respectively, and these
inhibitory activities were shown, using rat brush border membrane vesicles, to be competitive.
These results demonstrate that digestion is inhibited when an appropriate amount of ELM is
orally ingested with sucrose or polysaccharide in man. When ELM was orally administered in a
sucrose solution to fasted rats, the elevation in blood glucose was significantly suppressed,
depending on the concentration of ELM given. These results suggest that ELM could be used as
an ingredient in health foods and in foods that help to prevent diabetes.

Purification of total flavones from Morus alba L.by macroporous adsorbents and kinetic
model for the process.:Zhejiang Da Xue Xue Bao Yi Xue Ban. 2006 Mar;35(2):219-23.
Chinese.Chen JJ, Li XR, Fang X.College of Pharmaceutical Science, Zhejiang University,
Hangzhou 310031, China.

OBJECTIVE: To establish the separation and purification technology for the total flavones from
Morus alba L.(MTF), and present a kinetic model for this process. METHODS: Three types of
macroporous adsorbents were tested to separate and purify MTF, and to evaluated by adsorption
capacity, elution ratio and product purity. The curves of dynamic adsorption-elution process was
plotted and the kinetic equations were presented. RESULT: Among three types of macroporous
adsorbents, the NKA-9 type showed better property with the adsorption capacity of 43.4 mg.g(-
1)and the elution ratio of 98.2%. The purity of MTF product reached to 58.2 % in the
technological conditions following: extract sample concentration of 20 mg.ml(-1), 6 times
washing water and elution reagent of 70% ethanol. The adsorbent could be used for 4 times
repeatedly. The kinetic equation was y=u(1 - e(-kx)). CONCLUSION: The NKA-9 type of
macroporous absorbent is suitable to purify the MTF.

Screening of glucosidase inhibitors from various fractions of Mulberry leaves.:Zhongguo

Zhong Yao Za Zhi. 2006 Feb;31(3):223-7. Chinese.Yuan AH, Ma J, Jiang XF, Li S.Department
of Nephrology, Tongji Hospital of Tongji University, Shanghai 200065, China.
aihongyuanxn00@yahoo.com.cn

OBJECTIVE: To search for glucosidase inhibitors of various fractions extracted from mulberry
leaves. METHOD: The constituents of mulberry leaves water fraction were prepared by the
process of boiling, condensing, precipitating, exchanging with resins and rinsing. In vitro
glucosidase inhibitory activities were examined by photometric bioassay derived from rats. To
investigate in vivo effect of lowering blood glucose, the mouse blood glucose level was assayed
by glucose tolerance experiments. RESULT: The glucosidase inhibitory activities were found in
all the constituents of alkaloids, flavones and amyloses, the alkaloid constituent being the
strongest. CONCLUSION: The effect of reducing blood glucose of mulberry leaves is related to
the inhibitory activities against glucosidase of different constituents.

Enhancement of neuroprotection of mulberry leaves (Morus alba L.) prepared by the

anaerobic treatment against ischemic damage.:Biol Pharm Bull. 2006 Feb;29(2):270-4.Hiware
CJ.Department of Zoology, Dr. Babasaheb Ambedkar Marathwada University, Aurangabad
431004, MS, India. drhiware@rediffmail.com

Silk worm (Bombyx mori L.) larvae were fed on Mulberry leaves treated with Nux vomica
mother tincture. The impact on larval, cocoon, shell and pupal weight, silk ratio, average
filament length and denier, and number of breakages during reeling were investigated. The
results were positive in all parameters under study except cocoon weight, pupal weight, and the
average denier of the filament.

Mulberry latex rich in antidiabetic sugar-mimic alkaloids forces dieting on

caterpillars.:Proc Natl Acad Sci U S A. 2006 Jan 31;103(5):1337-41. Epub 2006 Jan 23.

Since ancient times, mulberry leaves (Morus spp.) have been used to rear the silkworm Bombyx
mori. Because the silkworm grows well on mulberry leaves, the toxicities and defensive
activities of these leaves against herbivorous insects have been overlooked. Here we show that
mulberry leaves are highly toxic to caterpillars other than the silkworm B. mori, because of the
ingredients of the latex, a milky sap exuded from mulberry leaf veins. The toxicity of mulberry
leaves was lost when the latex was eliminated from the leaves, and artificial diets containing
latex showed toxicity. Mulberry latex contained very high concentrations of alkaloidal sugar-
mimic glycosidase inhibitors reported to have antidiabetic activities, such as 1,4-dideoxy-1,4-
imino-D-arabinitol, 1-deoxynojirimycin, and 1,4-dideoxy-1,4-imino-D-ribitol. The overall
concentrations of these inhibitors in latex reached 1.5-2.5% (8-18% dry weight) in several
mulberry varieties, which were approximately 100 times the concentrations previously reported
from whole mulberry leaves. These sugar-mimic alkaloids were toxic to caterpillars but not to
the silkworm B. mori, indicating that the silkworm can circumvent the mulberry tree's defense.
Our results suggest that latex ingredients play key roles in defense of this tree and of other plants
against insect herbivory, and they imply that plant latexes are treasuries of bioactive substances
useful as medicines and pesticides.

Effects of herbal extracts on the function of human organic anion-transporting polypeptide

OATP-B.:Drug Metab Dispos. 2006 Apr;34(4):577-82. Epub 2006 Jan 13.

Most known interactions between herbal extracts and drugs involve the inhibition of drug-
metabolizing enzymes, but little is yet known about the possible role of transporters in these
interactions. In this study, we have examined the effects of herbal extracts used in dietary
supplements on the function of organic anion-transporting polypeptide B (OATP-B; OATP2B1),
which is expressed on human intestinal epithelial cells and is considered to be involved in the
intestinal absorption of various drugs. Specifically, the effects of 15 herbal extracts on uptake of
estrone-3-sulfate, a typical OATP-B substrate, by human embryonic kidney 293 cells stably
expressing OATP-B were evaluated. At concentration levels considered likely to be attainable in
the human intestine, extracts of bilberry, echinacea, green tea, banaba, grape seed, ginkgo, and
soybean potently inhibited estrone-3-sulfate uptake by 75.5, 55.5, 82.1, 61.1, 64.5, 85.4, and
66.8%, respectively (P < 0.01). The inhibitory effect of ginkgo leaf extract was concentration-
dependent (IC(50) = 11.2 +/- 3.3 microg/ml) and reversible. Moreover, flavonol glycosides and
catechins significantly inhibited the function of OATP-B, suggesting that the inhibitory effects of
the herbal extracts on OATP-B may be primarily attributable to flavonoids. The extracts of
mulberry, black cohosh, and Siberian ginseng moderately (but significantly) inhibited estrone-3-
sulfate uptake by 39.1, 47.2, and 49.2%, respectively (P < 0.05). Extracts of barley, Job's tears,
rutin, rafuma, and passionflower were ineffective. These results suggest that coadministration of
some dietary supplements may decrease the absorption of orally administered substrates of
OATP-B.

Single primer amplification reaction methods reveal exotic and indigenous mulberry
varieties are similarly diverse.:J Biosci. 2005 Dec;30(5):669-77.Bhattacharya E, Dandin SB,
Ranade SA.Plant Molecular Biology (Genomics), National Botanical Research Institute,
Lucknow 226 001, India.

Mulberry is the sole food source for mulberry silkworm and a number of indigenous and exotic
varieties are used in sericulture. Studies on assessment of genetic diversity have been done
amongst a few mulberry varieties using one or at the most two methods. However, no
comprehensive study on a large number of varieties has been carried out. In present study, single
primer amplification reaction (SPAR) methods have been used for determination of diversity in
27 mulberry varieties (exotic as well as indigenous), using four minisatellite core sequence
primers for directed amplification of minisatellite DNA (DAMD), three simple sequence repeat
(SSR) motifs as primers for inter simple sequence repeat (ISSR) and 20 arbitrary sequence
decamer primers for ran-dom amplified polymorphic DNA (RAPD) reactions. The Jaccard
coefficients were determined for the DAMD, ISSR and RAPD band data (total of 58, 39 and 235
bands respectively). All three methods revealed wide range of distances supporting a wide range
of mulberry genetic diversity. A cumulative analysis of the data generated by three methods
resulted in a neighbour-joining (NJ) tree that gave a better reflection of the relatedness and
affinities of the varieties to each other. Comparison of the three methods by marker indices and
the Mantel test of correlation indicated that though all methods were useful for the assessment of
diversity in mulberry, the DAMD method was better. When considered as two groups (10 exotic
and 17 indigenous varieties), the mulberry varieties in the exotic group were found to have
slightly greater diversity than the indigenous ones. These results support the concept of
naturalization of mulberry varieties at locales distant from their origins.

Antioxidant responses to enhanced generation of superoxide anion radical and hydrogen

peroxide in the copper-stressed mulberry plants.:Planta. 2006 May;223(6):1145-53. Epub 2005
Nov 15.Tewari RK, Kumar P, Sharma PN.Department of Botany, University of Lucknow,
226007 Lucknow, India. rajesh_bot@rediffmail.com
 The aim of the study was to implicate the generation of reactive oxygen species (ROS) and
altered cellular redox environment with the effects of Cu-deficiency or Cu-excess in mulberry
(Morus alba L.) cv. Kanva 2 plants. A study of antioxidative responses, indicators of oxidative
damage and cellular redox environment in Cu-deficient or Cu-excess mulberry plants was
undertaken. While the young leaves of plants supplied with nil Cu showed chlorosis and necrotic
scorching of laminae, the older and middle leaves of plants supplied with nil or 0.1 microM Cu
showed purplish-brown pigmented interveinal areas that later turned necrotic along the apices
and margins of leaves. The Cu-excess plants showed accelerated senescence of the older leaves.
The Cu-deficient plants showed accumulation of hydrogen peroxide and superoxide anion
radical. The accumulation of hydrogen peroxide was strikingly intense in the middle portion of
trichomes on Cu-deficient leaves. Though the concentration of total ascorbate increased with the
increasing supply of Cu, the ratio of the redox couple (DHA/ascorbic acid) increased in Cu-
deficient or Cu-excess plants. The activities of superoxide dismutase (EC 1.15.1.1), catalase (EC
1.11.1.6), peroxidase (EC 1.11.1.7), ascorbate peroxidase (EC 1.11.1.11) and glutathione
reductase (EC 1.6.4.2) increased in both Cu-deficient and Cu-excess plants. The results suggest
that deficiency of Cu aggravates oxidative stress through enhanced generation of ROS and
disturbed redox couple. Excess of Cu damaged roots, accelerated the rate of senescence in the
older leaves, induced antioxidant responses and disturbed the cellular redox environment in the
young leaves of mulberry plants.

Antimicrobial activity of 2-arylbenzofurans from Morus species against methicillin-

resistant Staphylococcus aureus.:Fitoterapia. 2005 Dec;76(7-8):708-11. Epub 2005 Oct 19.

Nine 2-arylbenzofurans isolated from Morus species were tested for their antimicrobial activities
against methicillin-sensitive Staphylococcus aureus (MSSA), methicillin-resistant S. aureus
(MRSA), Micrococcus luteus, Bacillus subtilis, Escherichia coli, Klebsiella pneumoniae and
Pseudomonas aeruginosa. Among these compounds, chalcomoracin (a leaf phytoalexine of
mulberry tree) exhibited considerable antibacterial activity against MRSAs (MICs 0.78 mug/ml).

Effects of Morus alba leaf extract on the production of nitric oxide, prostaglandin E2 and
cytokines in RAW264.7 macrophages.:Fitoterapia. 2005 Dec;76(7-8):608-13. Epub 2005 Oct
17.Choi EM, Hwang JK.Department of Biotechnology and Bioproducts Research Center, Yonsei
University, 134, Sinchon-dong, Seodaemun-gu, Seoul, 120-749, Korea.

Morus alba leaf methanolic extract and its fractions (chloroform, butanol, and aqueous fractions)
were found to inhibit NO production in LPS-activated RAW264.7 macrophages without an
appreciable cytotoxic effect at concentration from 4 to 100 microg/ml. LPS-induced PGE2
production was significantly reduced only by butanol fraction. In addition, M. alba leaf extract
and its fractions significantly decreased the production of TNF-alpha. These findings suggest
that M. alba leaf extract seems to be able in suppressing inflammatory mediators. Moreover, the
inhibitory activities on COX-2 and iNOS of its butanol fraction are warranted for further
elucidation of active principles for development of new antiinflammatory agents.
 Sterols of mulberry leaves and small leaf curl disease:Prikl Biokhim Mikrobiol. 2005 Jul-
Aug;41(4):460-2. Russian.Zambakhidze NE, Sulaberindze KV, Mzhavanadze VV, Tsiklauri
GCh.

Free and bound sterols of leaves of five mulberry cultivars differing in their susceptibility to
small leaf curl disease have been studied. The total content of sterols in all samples is similar and
is not correlated with the resistance of the cultivars. The qualitative composition of particular
sterols is also identical. They are represented by cholesterol, campesterol, stigmasterol, sitosterol,
and two 4alpha-methylsterols. The leaves of the most sensitive cultivar are characterized by high
cholesterol content. The ratio sitosterol : stigmasterol decreased in proportion to the resistance
level of a cultivar.

Asymmetrical introgression between two Morus species (M. alba, M. rubra) that differ in
abundance.:Mol Ecol. 2005 Oct;14(11):3471-83.Burgess KS, Morgan M, Deverno L, Husband
BC.Department of Botany, University of Guelph, Guelph, ON, Canada, N1G 2W1.
burgessk@virginia.edu

Asymmetrical introgression is an expected genetic consequence of hybridization when parental

taxa differ in abundance; however, evidence for such effects in small populations is scarce. To
test this prediction, we estimated the magnitude and direction of hybridization between red
mulberry (Morus rubra L.), an endangered species in Canada, and the introduced and more
abundant white mulberry (Morus alba L.) using nuclear (randomly amplified polymorphic DNA)
and cytoplasmic (chloroplast DNA sequence) markers. Parentage of 184 trees (n = 42 using
cpDNA) from four sympatric populations was estimated using a hybrid index and related to six
morphological characters and population frequencies of the parental classes. Overall, the
frequency of nuclear hybrids was 53.7% (n = 99) and ranged from 43% to 67% among
populations. The parental and hybrid taxa differed with respect to all of the morphological traits.
Sixty-seven percent of all hybrids contained more nuclear markers from M. alba than M. rubra
(hybrid index x = 0.46); among populations, the degree of M. alba bias was correlated with the
frequency of M. alba. In addition, the majority of hybrids (68%) contained the chloroplast
genome of white mulberry. These results suggest that introgression is bidirectional but
asymmetrical and is related, in part, to the relative frequency of parental taxa.

Determination of 1-deoxynojirimycin in leaves of Morus alba by high performance liquid

chromatography with fluorescence detection.:Zhongguo Zhong Yao Za Zhi. 2005
May;30(9):682-5. Chinese.Ouyang Z, Li YH, Xu WD, Chen J.School of Biological and
Environmental Engineering, Jiangsu University, Zhenyiang 212013, China.

OBJECTIVE: To develop a high performance liquid chromatographic method for the

determination of DNJ in Morus alba leaves with fluorimetric detection after precolumn
derivatization with 9-fluorenylmnethyl chlorformate (FMOC-Cl). METHOD: DNJ in Morus alba
leaves was extracted with 0.05 mol x L(-1) HCl, reacted with FMOC-Cl, and separated on a
HiQSiL C18 column at 250 degrees C. Mobile phase consisted of acetonitrile-0.1% aqueous
acetic acid (55:45) with a flow rate of 1.0 mL x min. The fluorescence detector was operated at
lambdaEX = 254 nm, lambdaEM = 322 nm. RESULT: A satisfactory separation between DNJ
and impurity was obtained. The calibration curve was linear over the concentration range from
0.567 microg x mL(-1) to 34 microg mL(-1), r = 0.9998. The average recovery was 97.2%. The
contents of DNJ in M. alba collected in different seasons and grown in different environment
were determined. CONCLUSION: The quantity of DNJ in leaves of M. alba is related to the
environment factor, temperature and growing period. The method can be used for qualioy control
of the medicinal material.

Absorption of mulberry root urease to the hemolymph of the silkworm, Bombyx mori.:J
Insect Physiol. 2005 Sep;51(9):1055-61. Epub 2005 Jul 6.

Mulberry leaves are the sole diet of the silkworm, Bombyx mori. The host urease is incorporated
into the larval hemolymph and involved in nitrogen metabolism in the insect. To investigate the
selective absorption of the host urease to the larvae, crude urease was prepared from mulberry
leaves and roots. Root urease was identical to leaf urease on the basis of electrophoretic analyses:
(1) the urease activity appeared in the same migration position in a native gel; (2) There was no
difference in molecular mass of the subunit. The root urease was orally injected to the fifth instar
larvae of the silkworm. Just before spinning, the larvae absorbed intact urease from the midgut
lumen to the hemolymph without the loss of activity. The capacity to absorb urease occurred
only at the specific stage. Localization of host urease in midgut tissue was observed using
confocal laser scanning microscopy and transmission electron microscopy. Based on spatial
distribution of immunofluorescent signals and immunogold particles, host urease specifically
attached to the surfaces of microvilli existing in the apical side of columnar cells and appeared in
the cytoplasm of the cells for transport to the hemolymph. The incorporation efficiency of root
urease into the hemolymph was significantly higher than for ureases from jack bean seeds and
Bacillus pasteurii. The urease that was transported to the hemolymph was electrophoretically
altered, compared with the host urease extracted.

Effects of dietary mulberry, Korean red ginseng, and banaba on glucose homeostasis in
relation to PPAR-alpha, PPAR-gamma, and LPL mRNA expressions.:Life Sci. 2005 Nov
12;77(26):3344-54. Epub 2005 Jun 23.Park MY, Lee KS, Sung MK.Department of Food and
Nutrition, College of Human Ecology, Sookmyung Women's University, Seoul, 140-742, Korea.

Despite lack of scientific evidences to support its therapeutic efficacy, the use of herbal
supplements has significantly increased. The purpose of this study was to evaluate the effects of
traditional anti-diabetic herbs on the progress of diabetes in db/db mice, a typical non-insulin-
dependent model. Five different experimental diets were as follows: control diet, 0.5% mulberry
leaf water extract diet, 0.5% Korean red ginseng diet, 0.5% banaba leaf water extract diet, and
0.5% combination diet (mulberry leaf water extract/Korean red ginseng/banaba leaf water
extract, 1:1:1). Blood levels of glucose, insulin, HbA1c, and triglyceride were measured every 2
weeks. At 12 weeks of age, animals were sacrificed, and tissue mRNA levels of PPAR-alpha,
PPAR-gamma, and LPL were determined. Results indicated that mulberry leaf water extract,
Korean red ginseng, banaba leaf water extract, and the combination of above herbs effectively
reduced blood glucose, insulin, TG, and percent HbA1c in study animals (p<0.05). We also
observed that the increased expressions of liver PPAR-alpha mRNA and adipose tissue PPAR-
gamma mRNA in animals fed diets supplemented with test herbs. The expression of liver LPL
mRNA was also increased with experimental diets containing herbs. The efficacy was highest in
animals fed the combination diet for all of the markers used. These results suggest that mulberry
leaf water extract, Korean red ginseng, banaba leaf water extract, and the combination of these
herbs fed at the level of 0.5% of the diet significantly increase insulin sensitivity, and improve
hyperglycemia possibly through regulating PPAR-mediated lipid metabolism.

Mulberry (Morus alba L.) leaves and their major flavonol quercetin 3-(6-
malonylglucoside) attenuate atherosclerotic lesion development in LDL receptor-deficient
mice.:J Nutr. 2005 Apr;135(4):729-34.

The effects of dietary consumption of mulberry (Morus alba L.) leaves and their major flavonol
glycoside, quercetin 3-(6-malonylglucoside) (Q3MG), on the development of atherosclerotic
lesions, in relation to the susceptibility of plasma LDL to oxidative modification, was studied in
LDL receptor-deficient (LDLR-/-) mice. Male mice aged 8 wk were randomly assigned to 4
groups (control, quercetin, Q3MG, and mulberry). The control group was fed an atherogenic-diet
containing 3 g cholesterol and 15 g cocoa butter/100 g. The other experimental groups were fed
the same atherogenic diet supplemented with 0.05 g quercetin/100 g for the quercetin group, 0.05
g Q3MG/100 g for the Q3MG group, and 3 g dried mulberry-leaf powder/100 g for the mulberry
group. The mice were fed their respective diets for 8 wk. The susceptibility of LDL to oxidative
modification was significantly decreased in the Q3MG- and mulberry-treated mice, as evidenced
by the 44.3 and 42.2% prolongation of the lag phase for conjugated diene formation compared
with that of the control mice. The atherosclerotic lesion area in both the Q3MG- and mulberry-
treated mice was significantly reduced by 52% compared with that of the controls. However, in
the quercetin group, no protective effects were observed against LDL oxidation or
atherosclerotic lesion formation. In conclusion, mulberry leaves attenuated the atherosclerotic
lesion development in LDLR-/- mice through enhancement of LDL resistance to oxidative
modification, and these antioxidative and antiatherogenic protective effects were attributed
mainly to Q3MG, the quantitatively major flavonol glycoside in mulberry leaves.

Simple and rapid determination of 1-deoxynojirimycin in mulberry leaves.:Biofactors.

2004;22(1-4):341-5.

A simple and rapid method for determining 1-deoxynojirimycin (DNJ), a potent glucosidase
inhibitor present in mulberry leaves (Morus alba and Morus bombysis), by high performance
liquid chromatography coupled to an evaporative light scattering detector (ELSD) has been
developed. DNJ was separated from extract of mulberry leaves on TSK gel Amide-80 column,
which is a representative column for hydrophilic interaction chromatography. During post
column detection, DNJ was detected by ELSD and concurrently identified by mass spectrometry.
The detection limit was 100 ng. This method is sufficiently sensitive for determining DNJ in
mulberry leaves and other related products.
 Effects of combination of Caiapo with other plant-derived substance on anti-diabetic
efficacy in KK-Ay mice.:Biofactors. 2004;22(1-4):149-52.

B. Ludvik et al., have recently shown the effect of Caiapo (Ipomoea batatas L.) on reducing
fasting blood glucose and insulin resistance in type-2 diabetic patients. It, however, was required
2-4 weeks after the single administration of Caiapo. The present study aimed to determine if the
combination therapy of Caiapo with a mulberry leaf powder, which inhibits alpha-glucosidase, or
with a loquat leaf extract, which shows an insulin-like effect, could make it possible to enhance
the antidiabetic activities of Caiapo, and to shorten the time necessary for the inhibition of
increasing blood glucose levels. A mixture of the pulverized tuber of Caiapo (357 mg/kg) and the
mulberry leaf powder (143 mg/kg), or a mixture of the pulverized skin of Caiapo (194 mg/kg)
and the powdered loquat leaf extract (6 mg/kg) was orally administered to 6 weeks-old male KK-
Ay mice for 28 days and the glucose loading test was conducted every 7 days. In the glucose
loading test after one week feeding, a reduction in blood glucose concentration after 60 minutes
of the administration of glucose was observed in both mixture groups against the control group
(p < 0.05) in the case of Caiapo only, similar delayed effects were seen in 2-3 weeks after
feeding.

Effects of hypervitaminosis of vitamin B3 on silkworm biology.:J Biosci. 2004

Dec;29(4):417-22. Etebari K, Matindoost L.Department of Sericulture, Faculty of Natural
Resources, University of Guilan, Somehe Sara, Iran. etebari@guilan.ac.ir

A high-dose of vitamin B(3) in silkworm diet interrupts larval feeding and normal growth. High
mortality of larvae occurs during molting and they cannot complete this process normally. Also
the larvae exhibit nicotinamide hypervitaminosis symptoms such as immobility, dyspepsia,
darkening of the skin, inability to excrete normally, exerting brownish fluid from anus and
swelling of rectal muscles. Maximum larval weights in 1, 2 and 3 g/l treatments were 2.9, 1.6
and 1.2 g respectively, while maximum larval weight in the control was 5.6 g. Larval stage
compared to control had increased 18, 26 and 31 days respectively. The concentration increase of
uric acid in haemolymph demonstrates the hyperuricemia, while other measured biochemical
compounds show significant decrease; sodium and potassium did not change significantly.

Inhibitory effects of mulberry leaf extract on postprandial hyperglycemia in normal rats.:J

Nutr Sci Vitaminol. 2004 Jun;50(3):161-4.

We examined the inhibitory effects of aqueous ethanol extract from mulberry leaves (ME) on
postprandial hyperglycemia in normal Wistar rats. ME dose-dependently suppressed the
postprandial rise of blood glucose in rats, when ME (0.02-0.5 g/kg) was given 0.5 h before the
administration of carbohydrates such as sucrose, maltose and starch. The ME dose showing 50%
inhibition of the increment of blood glucose (ED50) was 0.11 g/kg for sucrose, 0.44 g/kg for
maltose, and 0.38 g/kg for starch. ME and its basic fraction (MB) containing 1-deoxynojirimycin
were assayed for their inhibitory effects (IC50) on disaccharidase derived from the small
intestine of rats. The IC50 value of ME was 3.2 microg/mL for sucrase, 10 microg/mL for
isomaltase, and 51 microg/mL for maltase. The IC50 value of MB was 0.36 microg/mL for
sucrase, 1.1 microg/mL for isomaltase, and 6.2 microg/mL for maltase. The IC50 value of 1-
deoxynojirimycin as the principle component in ME was 0.015 microg/mL for sucrase and 0.21
microg/mL for maltase, and this value was comparable to the IC50 of voglibose. The inhibitory
activity of ME in a-amylase was weak. These results suggest that ME strongly suppresses
postprandial hyperglycemia after carbohydrate loading by inhibiting the activity of
disaccharidases in the small intestine of rats.

Analysis of phylogenetic relationship among five mulberry (Morus) species using

molecular markers.:Genome. 2004 Jun;47(3):439-48.Vijayan K, Srivastava PP, Awasthi
AK.Seri-Biotech Research Laboratory, Central Silk Board, Bangalore, Karnataka, India.

Species identification in mulberry (Morus) continues to be a point of great debate among

scientists despite the number of criteria such as floral characters, wood, and leaf anatomical and
biochemical characters used to identify the species within this genus. However, no consensus
system of classification has emerged. Hence, an investigation was undertaken with inter-simple
sequence repeat (ISSR) and random amplified polymorphic DNA (RAPD) markers to find out
the possibility of using these DNA markers to confirm the identity of genotypes in a particular
species. Fifteen ISSR and 15 RAPD primers generated 86% and 78% polymorphism,
respectively, among 19 mulberry genotypes. The polymorphism among the species varied from
50% to 57% in ISSR markers and 31% to 53% in RAPD markers. Similarity coefficients were
higher among the genotypes of M. latifolia, M. bombycis and M. alba. Cluster analyses separated
genotypes of M. laevigata and M. indica from those of the other species. Population structure
analysis of these species further showed high genetic differentiation coefficients (GST), high
heterozygosity between two species (DST), and total heterozygosity among populations (Ht)
coupled with considerably low gene flow (Nm) when M. laevigata was paired with other species.
Based on these parameters and the result of cluster analysis it is concluded that M. laevigata can
be considered as a separate species of mulberry, whereas the other four species may be grouped
together and treated as subspecies.

Determination of 1-deoxynojirimycin in mulberry leaves using hydrophilic interaction

chromatography with evaporative light scattering detection.:J Agric Food Chem. 2004 Mar
24;52(6):1415-8.

A simple and rapid method for determining 1-deoxynojirimycin (DNJ), a potent glucosidase
inihibitor present in mulberry leaves (Morus alba and Morus bombysis), by high-performance
liquid chromatography coupled to an evaporative light scattering detector (ELSD) has been
developed. DNJ was separated from an extract of mulberry leaves on a TSKgel Amide-80
column, which is a representative column for hydrophilic interaction chromatography. During
postcolumn detection, DNJ was detected by ELSD and concurrently identified by mass
spectrometry. The detection limit was 100 ng. This method is sufficiently sensitive for
determining DNJ in mulberry leaves and other related products.

Antioxidant role of mulberry (Morus indica L. cv. Anantha) leaves in streptozotocin-

diabetic rats.:Clin Chim Acta. 2003 Dec;338(1-2):3-10.Andallu B, Varadacharyulu
NCh.Department of Home Science, Sri Sathya Sai Institute of Higher Learning, Anantapur-
515001, India. andullusss@rediffmail.com

BACKGROUND: The antihyperglycemic and antioxidant role of mulberry (Morus indica L.)
leaves were investigated. METHODS: Streptozotocin (STZ)-induced diabetic male Wistar rats
were used as experimental models; one group was given 25% dry mulberry leaf powder mixed
with the standard diet and another group was given standard diet for a period of 8 weeks. The
antihyperglycemic and antioxidant role of mulberry was assessed by determining its effect on
blood glucose, lipid peroxidation, reduced glutathione (GSH) concentrations and on the activity
of glucose-6-phosphate dehydrogenase (G6PDH) and various antioxidant enzymes in
erythrocytes and compared with that of controls. RESULTS: Mulberry-treated diabetic rats
showed a significant decrease in fasting blood glucose concentrations indicating a good glycemic
control. Increased lipid peroxidation and the activity of catalase (CAT) in erythrocytes observed
in diabetic controls were significantly decreased by mulberry leaves (48% and 33%,
respectively). Decreased GSH concentrations and the activity of glucose-6-phosphate
dehydrogenase and antioxidant enzymes viz., glutathione peroxidase (GPx), glutathione
reductase (GR), glutathione-S-transferase (GST) and superoxide dismutase (SOD) observed in
uncontrolled diabetes were improved (52%, 69%, 151%, 95%, 24% and 106%) by mulberry
treatment very efficiently. CONCLUSION: Mulberry leaves possess antihyperglycemic and
antioxidant properties.

Ninety-day dietary toxicity study of mulberry leaf extract in rats.:Shokuhin Eiseigaku

Zasshi. 2003 Aug;44(4):191-7.

Mulberry leaf extract was studied toxicologically in male and female SD rats. The extract was
administered orally at concentrations of 0% (control group), 0.1%, 0.4% and 1% in basal diet for
90 days. No remarkable change in test animals of both sexes was observed in terms of body
weight gain or at necropsy. Hematology and blood chemistry revealed no abnormalities.
Pathological examination revealed no toxic change in any organ observed. These findings
indicate that dietary intake of 1% mulberry leaf extract for 90 days (884.5 mg/kg/day for males
and 995.7 mg/kg/day for females as mean daily intake) causes no toxicological change in rats.

Mulberry (Morus alba) leaves as human food: a new dimension of sericulture.:Int J Food
Sci Nutr. 2003 Nov;54(6):411-6.Srivastava S, Kapoor R, Thathola A, Srivastava RP.Department
of Foods and Nutrition, College of Home Science Govind Ballabh Pant University of Agriculture
and Technology, U.S. Nagar, Uttaranchal, India.

Mulberry leaf is commonly used for sericulture in almost every part of the world but its
potential to be utilized for human consumption is not well recognized. This paper deals with
development of mulberry leaf powder and its use with wheat flour to develop paratha, the most
common food item of breakfast and dinner in the Indian diet. The optimum ratio of the mulberry
leaf powder and wheat flour (MLP-WF) mix for preparation of paratha on the basis of sensory
quality was found to be 1:4. The protein quality of the MLP-WF mix was estimated by
measuring the Protein Efficiency Ratio, and was found to be 1.82 against a casein diet for which
a value of 2.44 was observed. The in vivo toxic effect of mix was studied and no adverse effect
on the growth of internal organs of rats (heart, liver, kidney and testes) was found. The storage
stability of the mix was estimated for a period of 2 months in polyethylene bags at room
temperature. A non-significant difference was observed between paratha prepared from fresh and
stored mix. This indicated that mix can be stored for a period of 2 months at room temperature
without loss of quality.

Determination of 1-deoxynojirimycin in Morus alba L. leaves by derivatization with 9-

fluorenylmethyl chloroformate followed by reversed-phase high-performance liquid
chromatography.:J Chromatogr A. 2003 Jun 20;1002(1-2):93-9.Kim JW, Kim SU, Lee HS, Kim
I, Ahn MY, Ryu KS.Department of Sericulture and Entomology, National Institute of
Agricultural Science and Technology, Suwon 441-100 South Korea. annua38@empal.com

A rapid and reliable method suitable for assays of a large number of Morus alba leaves for 1-
deoxynojirimycin (DNJ) has been developed. DNJ in 0.1 g of freeze-dried leaves was double-
extracted in 10 mL of aqueous 0.05 M HCl by vortexing for 15 s at room temperature,
derivatized with 9-fluorenylmethyl chloroformate (FMOC-Cl), and analyzed by reversed-phase
high-performance liquid chromatography (RP-HPLC) equipped with a fluorescence detector.
The double extraction recovered > 99% of extractable DNJ from the leaves. Stabilization of
FMOC-derivatized DNJ (DNJ-FMOC) was achieved by diluting the reactant with aqueous acetic
acid after derivatization. DNJ-FMOC was stable for at least 16 days under acidic conditions at
room temperature (24 degrees C). Linearity ranged between 0.3 and 30 microg mL(-1). The
intra- and inter-day precision for DNJ-spiked biological samples was between 0.6 and 1.8% and
between 3.7 and 4.5%, respectively.

Quantitative analysis of aglycone quercetin in mulberry leaves (Morus alba L.) by

capillary zone electrophoresis.:Electrophoresis. 2003 Apr;24(7-8):1236-41.Suntornsuk L,
Kasemsook S, Wongyai S.Department of Pharmaceutical Chemistry, Faculty of Pharmacy,
Mahidol University, Rajathevee, Bangkok, Thailand. pylll@mahidol.ac.th

A capillary zone electrophoresis method was established for analysis of aglycone quercetin in
mulberry leaves (Morus alba L.). The influence of, e.g., background electrolyte concentrations
and pH, surfactant concentrations, organic solvents, temperature, and voltage on the separation
of aglycone quercetin, rutin, quercitrin, kaempferol, catechin, and gallic acid was systematically
investigated. The optimum condition providing baseline separation of all compounds within 16.5
min was obtained in 150 mM boric acid (pH 10.0) using a fused-silica capillary with an effective
length of 42.5 cm (50 microm inner diameter), temperature of 32 degrees C, and voltage of 15
kV. Method assessment was performed by standard addition method using rutin as an internal
standard. Linearity of the method was excellent (r(2) > 0.999) over the concentration tested (40-
160 microg/mL). The relative standard deviations (%RSDs) from injection, intraday, and
interday precision were less than 2.5%. Recoveries were good (asymptotically equal to 100.0%,
%RSD = 0.8%) with a limit of detection (LOD) and limit of quantitation (LOQ) of 0.86 and 3.16
microg/mL (%RSD = 1.8%), respectively. The aglycone quercetin found in the mulberry leaves
was 0.452 g/100 g (%RSD = 0.6%) on dry weight.
 Control of hyperglycemia and retardation of cataract by mulberry (Morus indica L.) leaves
in streptozotocin diabetic rats.:Indian J Exp Biol. 2002 Jul;40(7):791-5.Andallu B,
Varadacharyulu NCh.Department of Home Science, Sri Sathya Sai Institute of Higher Learning,
Anantapur 515001, India. andallusss@rediffmail.com

Dried leaf powder of mulberry (M. indica L.) when given along with the diet at 25% level to
streptozotocin induced diabetic male Wistar albino rats for 8 weeks, controlled hyperglycemia,
glycosuria, albuminuria and retarded onset of retinopathy. Untreated diabetic rats showed
hyperglycemia, glycosuria, albuminuria and developed lenticular opacity after 8 weeks of
experimental period.

Microscopic identification on the Folium Mori and the leaves of its allied species from
Shandong.:Zhong Yao Cai. 1999 Dec;22(12):622-4. Chinese.Guo Q, Zhou F, Zhang X.Shandong
University of TCM, Jinan 250014.

The Folium Mori and the leaves of its allied species Morus australis Poir., M. mongolica
Schneid. from Shandong were identified. The result shows that they are identified easily and
accurately according to epidermis, trichomes and blade.

Determination of rutin and quercetin in mulberry leaves by high performance capillary

electrophoresis.:Se Pu. 2001 Sep;19(5):395-7. Chinese.Sun L, Meng L, Chen J, Ma J, Hu R, Jia
DZ.Department of Chemistry, Xinjiang Medical University, Wulumuqi 830054, China.

Rutin and quercetin are the main effective components of mulberry leaves with the functions of
controlling the increase of fat in serum and controlling the formation of arterio-sclerosis. In this
article a high performance capillary electrophoretic(HPCE) method was used to separate and
determine rutin and quercetin in mulberry leaves collected from different periods, different
places and different kinds in Xinjiang. Electrophoretic conditions were as follows: a capillary
tube (75 microns i.d. x 57 cm (effective length, 50 cm)), with 10 mmol/L dihydrogen sodium
phosphate-20 mmol/L sodium borate containing 15% methanol as the running buffer (pH 8.62)
and an applied voltage of 20 kV, at 25 degrees C, detected at a wavelength of 245 nm. Under the
optimum conditions, rutin and querctin were separated successfully from other components
within 12 minutes. The corrected peak areas of rutin and quercetin increased linearly with the
increase of their concentrations in the range of 4.4 mg/L-28.8 mg/L and 1.8 mg/L-145.8 mg/L
respectively. The corresponding regression equations for rutin and quercetin were Y = 71.60 rho
+ 30.51 (r = 0.9991) and Y = 315.17 rho - 929.70 (r = 0.9949) respectively, and the recoveries
were 95.64% and 99.36%. The analytical results demonstrate the method is simple, quick and
well reproducible, and can be used as a reliable tool for the quality control of mulberry leaves.

Inoculation of field-established mulberry and papaya with arbuscular mycorrhizal fungi

and a mycorrhiza helper bacterium.:Mycorrhiza. 2002 Dec;12(6):313-6. Epub 2002 Sep
24.Mamatha G, Bagyaraj DJ, Jaganath S.Department of Agricultural Microbiology, University
of Agricultural Sciences, GKVK Campus, Bangalore 560 065, Karnataka, India.

The effects of soil inoculation with arbuscular mycorrhizal (AM) fungi and a mycorrhiza helper
bacterium (MHB) were investigated on mulberry and papaya plants already established in the
field. Ten-year-old mulberry plants (var. M5) were inoculated with Glomus fasciculatum and
1.5-year-old papaya plants (var. Solo) were inoculated with a mixed culture of G. mosseae and
G. caledonium with or without Bacillus coagulans at two levels of P fertilizer. Growth, P uptake,
yield and AM colonization levels were monitored. Leaf yield in mulberry and fruit yield in
papaya were minimal in uninoculated plants given 50% recommended P. However, crop yields
of both mulberry and papaya inoculated with AM fungi alone or together with the bacterium and
given 50% recommended P were statistically on a par with that of uninoculated plants given
100% recommended P. As inoculation of B. coagulans increased mycorrhiza levels in AM
fungal-inoculated plants, this may be included in the class of MHB. Thus, mulberry and papaya
already established in the field may respond to AM inoculation and MHB may increase
symbiosis development by efficient AM fungi.

Administration of Folium mori extract decreases nitric oxide synthase expression in the
hypothalamus of streptozotocin-induced diabetic rats.:Jpn J Pharmacol. 2002 Oct;90(2):189-
92.Jang MH, Kim H, Shin MC, Lim BV, Lee TH, Jung SB, Kim CJ, Kim EH.Department of
Physiology, College of Medicine, Kyung Hee University, Seoul, Korea.

Folium mori, the leaves of Morus alba L., has traditionally been used for the treatment of
diabetic hyperglycemia. It has been shown to induce enhanced NOS expression in the
hypothalamus of rats with streptozotocin (STZ)-induced diabetes. In the present study, the effect
of Folium mori on the expression of nitric oxide synthase (NOS) in the hypothalamus of STZ-
induced diabetic rats was investigated via nicotinamide adenine dinucleotide phosphate-
diaphorase (NADPH-d) histochemistry. Enhanced NAPDH-d expression was detected in the
paraventricular nucleus, ventromedial hypothalamic nucleus, and lateral hypothalamic area of the
hypothalamus in the STZ-induced diabetes group. Administration of the aqueous extract of Fo

lium mori to rats with STZ-induced diabetes resulted in decreased NADPH-d positivity. These
results suggest that Folium mori treatment is effective in curbing the desire for food under
diabetic conditions via modulation of NO expression in the hypothalamus.

Mulberroside F isolated from the leaves of Morus alba inhibits melanin biosynthesis.:Biol
Pharm Bull. 2002 Aug;25(8):1045-8.Lee SH, Choi SY, Kim H, Hwang JS, Lee BG, Gao JJ, Kim
SY.Department of Herbal Pharmacology, Graduate School of East-West Medical Science,
Kyunghee University, Seoul, Korea.

The current study was carried out to investigate the in vitro effects of an 85% methanol extract
of dried Morus alba leaves on melanin biosynthesis, which is closely related to
hyperpigmentation. These extracts inhibited the tyrosinase activity that converts dopa to
dopachrome in the biosynthetic process of melanin. Mulberroside F (moracin M-6, 3'-di-O-beta-
D-glucopyranoside), which was obtained after the bioactivity-guided fractionation of the
extracts, showed inhibitory effects on tyrosinase activity and on the melanin formation of melan-
a cells. This compound also exhibited superoxide scavenging activity that is involved in the
protection against auto-oxidation. But its activity was low and was weaker than of kojic acid.
These results suggest that mulberroside F isolated from mulberry leaves might be used as a skin
whitening agent.

Effect of mulberry (Morus indica L.) therapy on plasma and erythrocyte membrane lipids
in patients with type 2 diabetes.:Clin Chim Acta. 2001 Dec;314(1-2):47-53. Andallu B,
Suryakantham V, Lakshmi Srikanthi B, Reddy GK.Department of Home Science, Sri Sathya Sai
Institute of Higher Learning, Anantapur 515001, Andhra Pradesh, India.

BACKGROUND: Mulberry (Morus indica L.) is non-toxic natural therapeutic agent shown to
possess hypoglycemic, hypotensive, and diuretic properties. METHODS: The hypoglycemic
effect of the mulberry leaves was evaluated by comparing the anti-diabetic activity of the
standard drug, glibenclamide. A total of 24 type 2 diabetic patents were divided randomly into
two treatment groups: the mulberry agent and glibenclamide, for 30 days. Serum and erythrocyte
membrane lipid profiles of the patients were analyzed before and after the treatments.
RESULTS: Patients with mulberry therapy significantly improved their glycemic control vs.
glibenclamide treatment. The results from pre- and post-treatment analysis of blood plasma and
urine samples showed that the mulberry therapy significantly decreased the concentration of
serum total cholesterol (12%, p<0.01), triglycerides (16%, p<0.01), plasma free fatty acids (12%,
p<0.01), LDL-cholesterol (23%, p<0.01), VLDL-cholesterol (17%, p<0.01), plasma peroxides
(25%, p<0.01), urinary peroxides (55%, p<0.01), while increasing HDL-cholesterol (18%,
p<0.01). Although the patients with glibenclamide treatment showed marginal improvement in
glycemic control, the changes in the lipid profile were not statistically significant except for
triglycerides (10%, p<0.05), plasma peroxides (15%, p<0.05), and urinary peroxides (19%,
p<0.05). Both treatments displayed no apparent effect on the concentrations of the glycosylated
hemoglobin (Hb A(1)c) in diabetic patients. However, the fasting blood glucose concentrations
of diabetic patients were significantly reduced by the mulberry therapy. CONCLUSIONS:
Mulberry therapy exhibits potential hypoglycemic and hypolipidemic effects in diabetic patients.

Polyhydroxylated alkaloids isolated from mulberry trees (Morusalba L.) and silkworms
(Bombyx mori L.).:J Agric Food Chem. 2001 Sep;49(9):4208-13.

New polyhydroxylated alkaloids, (2R,3R,4R)-2-hydroxymethyl-3,4-dihydroxypyrrolidine-N-

propionamide from the root bark of Morus alba L., and 4-O-alpha-D-galactopyranosyl-
calystegine B(2) and 3 beta,6 beta-dihydroxynortropane from the fruits, were isolated by column
chromatography using a variety of ion-exchange resins. Fifteen other polyhydroxylated alkaloids
were also isolated. 1-Deoxynojirimycin, a potent alpha-glucosidase inhibitor, was concentrated
2.7-fold by silkworms feeding on mulberry leaves. Some alkaloids contained in mulberry leaves
were potent inhibitors of mammalian digestive glycosidases but not inhibitors of silkworm
midgut glycosidases, suggesting that the silkworm has enzymes specially adapted to enable it to
feed on mulberry leaves. The possibility of preventing the onset of diabetes and obesity using
natural dietary supplements containing 1-deoxynojirimycin and other alpha-glucosidase
inhibitors in high concentration is of great potential interest.

Mulberry moracins: scavengers of UV stress-generated free radicals.:Biosci Biotechnol

Biochem. 2001 Jun;65(6):1402-5.

Mulberry leaves treated with UV-C were found to accumulate three different phytoalexins,
moracin C, moracin N, and chalcomoracin. The increased level of malondialdehyde in UV-
treated leaves along with moracins suggested their role as a free-radical scavenger in stressed
plants. All the three moracins induced under UV stress were capable of scavenging the
superoxide anion generated by the xanthine-xanthine oxidase system. Also, moracins were
capable of inhibiting lipid peroxidation, which strongly indicates their role as a scavenger.

Mulberry leaf extract inhibits the oxidative modification of rabbit and human low density
lipoprotein.:Biol Pharm Bull. 2000 Sep;23(9):1066-71.

In a previous study, we demonstrated that the intake of mulberry leaves or their 1-butanol
extract (MLBE) reduced the concentration of serum lipids and atheromatous thickening of
arterial intima in hypercholesterolemic rabbits. In the present study, we investigated the
antioxidative activity of MLBE and isoquercitrin, the main component of MLBE. First, we
determined the effect on a stable radical agent, finding that quercetin, isoquercitrin and MLBE
scavenged the DPPH radical. We then determined the copper-induced oxidative modification of
rabbit and human low-density lipoprotein (LDL). Oxidation of LDL was spectrophotometrically
monitored by changes in absorbance at 234 nm accompanied by the formation of conjugated
dienes, and measured the formation of thiobarbituric acid reacting substances (TBARS).
Quercetin, an aglycone of isoquercitrin, inhibited the formation of conjugated dienes and
TBARS by copper-induced oxidative modification of rabbit and human LDLs. MLBE and
isoquercitrin also inhibited the oxidation of LDL. These results indicate that mulberry leaves
inhibit the oxidative modification of LDL and suggest that mulberry leaves may had prevent
atherosclerosis.

Purification and properties of urease from the leaf of mulberry, Morus

alba.:Phytochemistry. 2000 Feb;53(3):325-30.

Urease was purified from leaves of mulberry (Morus alba, L.) by ammonium sulfate
fractionation, acetone fractionation and sequential column chromatography including Q-
Sepharose HP, Phenyl-Sepharose HP, Superdex 200 HR and Mono Q. The enzyme was purified
5700-fold to apparent homogeneity with a recovery of 3.6%. The molecular mass of the enzyme
was determined to be 90.5 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis
analysis and 175 kDa by gel filtration, indicating that the enzyme was a homodimer. In the
western blot analysis, 90.5 kDa subunit of the mulberry leaf urease cross-reacted with antiserum
raised against jack bean seed urease. The N-terminal sequence of the first 20 residues of the
enzyme revealed that it has a high similarity (80-90%) to ureases from other plant sources,
suggesting that the mulberry leaf urease is closely related to other plant ureases. However, the
mulberry leaf enzyme showed an optimum pH for activity of 9.0, while the optimum pH of most
ureases isolated from plants and bacterial is neutral. In addition, the K(m) value for urea was
0.16 mM, which is lower than those of ureases from other sources. It is also proposed that urease
activity ingested by browsing silkworm releases ammonia that is subsequently used in silkworm
protein synthesis.

Variations in amounts of carbohydrates, amino acids and adenine nucleotides in mulberry

tree (Morus alba L.) stems during transitional phases of growth.:Tree Physiol. 1990
Jun;6(2):191-200.

Quantitative changes in carbohydrates, amino acids and adenine nucleotides in the stems of
mulberry trees (Morus alba L., cv. Shin-ichinose) were followed from spring to early summer
and from autumn to early spring. Both ATP and ADP content of stems increased before bud
break, whereas the content of sucrose, the most abundant sugar among the stem carbohydrates,
decreased. The sucrose content fell to its lowest value at the beginning of May, and then
increased rapidly, whereas the starch content decreased suggesting consumption of reserve
carbohydrate and simultaneous accumulation of current photosynthate. This was confirmed by
studies in which reserve carbohydrates were labeled with (14)CO(2). Proline content of stems
increased from the time of leaf shedding until early spring. Although it was the most abundant
amino acid at the time of bud break, proline rapidly decreased as new shoots developed and was
hardly detectable by the beginning of May. The asparagine and arginine contents increased
transiently following bud break, and then decreased toward summer. Transient increases in
glutamine and arginine were noted at the time of leaf shedding.

Comparison of Lodging Safety Factor of Untreated and Succinic Acid 2,2-

Dimethylhydrazide-Treated Shoots of Mulberry Tree.:Plant Physiol. 1990 Jan;92(1):12-16.

This study examined the lodging resistance of mulberry tree (Morus bombycis Koidz. cv
Kenmochi) shoots treated or not treated with succinic acid 2,2-dimethylhydrazide (SADH). The
lodging safety factor, an indicator of lodging resistance, was defined as the ratio of critical
lodging load to the leaf fresh weight observed, provided that the distribution of the critical
lodging load along the stem was similar to that of the leaf fresh weight observed. The critical
lodging load was experimentally estimated by loading weights onto the stems. In the untreated
trees, the lodging safety factor was maintained at about 3.2. In the SADH-treated trees, the stem
elongation was inhibited to about 80% of that in the untreated trees, and the percentage of shoot
dry matter partitioned into the leaves was always larger than that of the untreated trees. This
dwarfing of the stem caused by SADH increased the critical lodging load supported by the unit
stem dry weight, while this large investment of materials in leaves increased the leaf fresh weight
supported by the unit stem dry weight. Since the increments canceled each other, the lodging
safety factor of the SADH-treated shoots was similar to that of the untreated ones. These results
suggest that the shoot formation of the mulberry tree is controlled to maintain the lodging safety
factor at a constant level.