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 Folium 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.