Journal of Physiology & Pathology in Korean Medicine
/
v.22
no.3
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pp.684-691
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2008
As part of studies to develop new materials to lower blood glucose levels using crude polysaccharide, this study was attempted to analyze the characteristics of crude polysaccharide obtained from the extracts of a mixed herbal medium(OCM) where Trichloloma matsutake mycelium and Cordyceps militaris mycelium were cultured together and to look into the influence of administering these by concentration upon the blood glucose and serum lipid levels of rats with diabetes which was induced by STZ(Streptozotosin). Experimental group was divided into 6 groups: first, it was divided into normal control group(NC group) and diabetes-induced group, and diabetes-induced group was subdivided into diabetic control group(DC group), acarbose-treated group(PC group), 100 mg/kg/body weight-treated by crude polysaccharide of OCM(UE) group(UE100 group), 200 mg/kg/body weight-treated group(UE200 group), and 300 mg/kg/body weight-treated group(UE300 group). In diabetic-induced groups, after streptozotocin was melted in 0.01M citrate buffer at 50 mg/kg/body weight, when the non-fasting blood glucose level not on an empty stomach was 300 mg/dl or more in blood collected from the tail vein, it was regarded as diabetic induction and then such diabetic-induced experimental animals were used in this experiment. The yield of crude polysaccharide obtained from OCM was found to be 0.31% and the ${\beta}$-glucan content 39.40%. As a result of analyzing NO on immune function, which is known as major physiological activity of crude polysaccharide, high NO viability was shown; when 1 mg/ml LPS was treated at 1 ug/ml, it was found to be 50.77 uM, and when LPS was treated at 10 ug/m, it was found to be 53.78 uM. Also, regarding cancer cells, cell count was decreased by about 26% in proportion to sample concentration, while for normal cells, it was a little decreased in proportion to concentration, however, cell count was maintained in the range of $81.92{\sim}98.16%$ at all concentrations. In case of blood glucose level, it was decreased in all extract-treated groups compared to DC group and in the cases of ALT and AST, they were found to be lower in extract-treated groups compared to PC group and for serum lipid, it was found to be lower in UE100 group compared to PC group. Thus this study tried to utilize these results as fundamental data for development of preventive and therapeutic agents against diabetes as well as functional foods using the crude polysaccharide of mushrooms.
Inflammation scan using radiolabelled leukocytes has high sensitivity and specificity. Several methods for labelling leukocytes have been evaluated using P-32 diisopropyl fluorophosphate (DFP-32), H-3 thymidine, Cr-51 chromate, Ga-67 citrate and Tc-99m-sulfur colloid. In-111-oxine has proved so far to be the most reliable agent for labelling leukocytes. In-111-oxine is, however, expensive, not easily available when needed, and its radiation dose to leukocytes is relatively high. Moreover, resolution of the resultant image is relatively poor. Tc-99m is still the agent of choice because of, as compared with the indium, its favorable physical characteristics, lower cost and availability. Now the technique for labelling the leukocytes with technetium is successfully obtained using the lipophilic HAPAO with higher efficiency for granulocytes than for other cells. With this technique it is possible to label leukocytes in plasma to improve the viability of the leukocytes. Inflammation scan using Tc-99m-HMPAO has been evaluated in several laboratories, and difference in methods for separation and labelling accounts for difference in efficiency, viability and biodistribution of the labelled leukocytes. We performed inflammation scan using leukocytes labelled with Tc-99m-HMPAO in three dogs 24 hours after inoculation of live E. Coli and A. Aureus in their right abdominal wall. We separated mixed leukocytes by simple sedimentation using 6% hetastarch (HES) and labelled the leukocytes with Tc-99m-HMPAO in 20% cell free plama diluted with phosphate buffer solution(Fig. 1). Uptake was high in the liver and spleen but is was minimal in the lungs on whole body scan. Kidneys and intestine showed minimal activity although it was high in the urinary bladder(Fig. 2). Uptake of labelled leukocytes in the inflammation site was do(mite on 2 hour-postinjection scan and abscess was clearly delineated on 24 hour-delayed scan with high target-to-nontarget ratio(Fig. 3, 4). Inflammation scan using mixed leukocytes labelled with Tc-99m-HMPAO is very sensitive and specific in early detection of inflammation.
The metabolism of Entamoeba histolytica would be affected by various environmental factors, and alteration of the environment was known to afEect the fine structure of 5. histolytica. The present study was designed electronmicroscopically to investigate the ultrastructure and enzyme activities in the aEonic and conventional strains of 5. histolytica. The trophozoites of axenically cultivated HK-9 strain and conventional YS-27 and YS-49 strains of 5. histolytica were collected and liKed with 4% paraformaldehyde/0.1M cacodylate buffier(pH 74), After washing them by centrifugation, 1% warm agar was added in the sediment. Solidified agar with the trophozoites was cut into $lmm^3$ cubes, and incubated in the various substrates to observe enzyme activities. Then, the specimen was post-fixed with 3% glutaraldehyde/0.1M cacodylate buffer (PH 7.4) and 1% osmium tetroBide/0.1M cacodylate buffier (pH 7.4) , dehydrated in ascending ethanol series and embedded in epoxy resin. These were sectioned on an ultramicrotome and observed with a transmission electronmicroscope. The procedures for the observation of the fine structure were same as the above, except for the incubation in the substrate. The sections were stained with uranyl acetate and lead citrate. For the observation of the surface of the amoebae, scanning-electronmicroscopy was carried out. The results obtained in the present study are summarized as follows: 1. The fuzzy coat around double-layered plasma membrane of 5. histolytica was more irregularly and densely distributed in the conventional strains (YS-27, YS-49 strains) than in the axonic strain (HK-9 strain). 2. The endosomes, button bodies and chromatin material were surrounded by a double-layered nuclear membrane having scattered nuclear fores. The paranuclear body, mono- or double-layered vacuoles, vacuolar membrane whorls, rosette-like cylindrical bodies, aggregation of cylindrical bodies and helical bodies were found in the cytoplasm of the amoebae. Helical bodies and glycogen granules were generally abundant, while a few smooth endoplasmic reticula were observed in the cytoplasm. 3. Alkaline phosphatase activity was mainly demonstrated in the plasma membrane, limiting membranes of vacuoles and smooth endoplasmic reticula. ATPase activity was observed in the nucleus, limiting membranes of vacuoles and vacuolar membrane whorls. 4. Acid phosphatase activity was commonly demonstrated in the limiting membranes an contents of vacuoles, Iysosome-like organelles, plasma membrane and the button bodies in the nucleus. The activity was more weakly demonstrated in the HK-9 strain than in the other conventional strains of 5. histolytica. No peroBidase activity was observed in the amoeba strains employed in the present study. 5. With a scanning electron-microscope, no distinct structural differences were observed between the amoeba strains. All the trophozoite forms of the amoebae showed crater-like depressions and rugged features on the outer surface.
Journal of the Korean Society of Food Science and Nutrition
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v.27
no.2
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pp.319-325
/
1998
The purpose of this study was to investigate the effect of green tea catechin on microsomal mixed function oxidase(MFO) system of kidney and brain in streptozotocin(STZ) induced diabetic rats. Sprague-Dawley male rats weighing 140$\pm$10g were randomly assigned to one control and three STZ-diabetic groups. Diabetic groups wer classified to DM-0C(catechin 0%/kg diet), DM-0.5C (catechin 0.5%/kg diet), and DM-1.0C(catechin 1%/kg diet) according to the level of catechin supplementation. Diabetes were experimentally induced by intravenous administration of 55mg/kg body weight of STZ in citrate buffer(pH 4.3) after 4 weeks feeding of three experimental diets. Animals were sacrificed at the sixth day of diabetic state. The contents of cytochrome P450 in kidney were increased by 77, 42, 49% in DM-0C, DM-0.5C and DM-1.0C groups, respectively, than normal group. The contents of cytochrome P450 in brain were increased by 43% in DM-0C group than normal group, but those of DM-0.5C and DM-1.0C groups were similar to that of normal group. The contents of cytochrome b5 in kidney were increased by 78, 38, 49% in DM-0C, DM-0.5C and DM-1.0C groups, respectively, than normal group. Meanwhile, the contents of cytochrome b5 in brain were not significantly different among all groups. The activities of NADPH-cytochrome P450 reductase in kidney of DM-group were increased by 27% than normal group, but those of DM-0.5C and DM-1.0C groups were 13 and 15% lower than that of DM-0C group. The activities in brain were also increased by 31% in DM-0C group, but those of DM-0.5C and DM-1.0C groups were similar to than of normal group. Levels of TBARS (thiobarbituric acid reactive substance) in kidney were increased by 147, 60 and 59% in DM-0C, DM-0.5C, and DM-1.0C groups, respectively, compared with normal group, but those of DM-0.5C and DM-1.0C groups were 36, 35% lower than that of DM-0C group. Meanwhile, the levels of TBARS in brain were not significantly different among four groups. These results indicate that dietary catechins in green tea play a powerful antioxidant role in reducing the lipid peroxidation enhanced by activation of MFO system in STZ-induced diabetes.
This study was designed to investigate a natural medicinal multi-plants extract (BG515), which consisted of multi extracts of Mori folium, Rehmannia glutinosa Liboschitz, Dioscorea japonica, Lycii fructus, and Astragalus radix, on blood glucose, insulin levels, and serum malondialdehyde (MDA) concentrations in streptozotocin-induced diabetic rats. Streptozotocin (STZ) induces a type 1 diabetes mellitus in rats. Type 1 is usually characterized by the presence of islet cell autoantibodies (ICA), autoantibodies to insulin (IAA), and autoantiboides to glutamic acid decarboxylase (GAD), which identify the autoimmune process that leads to $\beta-cell$ destruction. Thirty-five male Sprague-Dawley (SD) rats weighing $150\sim170g$ each (6 weeks old) were randomly divided into one control (Group A) and 4 STZ-induced diabetic groups, and were subjected to one of the following treatment for 12 weeks. Groups A and B were fed basal diets and Group C, D, and E received the same diets as groups A and B, but with supplements of 150 mg/kg, 300 mg/kg, and 600 mg/kg of BG515 orally for 12 weeks, respectively. Diabetes was induced in Groups B, C, D, and E by intravenous injection of 45 mg/kg of STZ per body weight in sodium citrate buffer (pH 4.5) via the tail vein. In the BG515 groups, we found increases in serum insulin levels, compared to the STZ-control group, but these data were not significant. In contrast, blood glucose and serum MDA concentrations decreased in the BG515 groups compare to the STZ-control group. At the 5th week, in all the BG515 administered groups, there were decreases in serum blood glucose levels compared to the STZ- control group, and this activity was very strong in the BG515-1 group at the 12th week. These results suggest that natural bio-complex compounds (BG515) may slightly suppress STZ-induced changes in serum MDA concentration via the maintenance of serum insulin levels, due to the prevention of $\beta-cell$ and glucagon destruction by STZ.
Journal of the Korean Society of Food Science and Nutrition
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v.28
no.3
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pp.654-662
/
1999
The purpose of this study was to investigate the effects of vitamin E on the antioxidative defense system of kidney in streptozotocin induced diabetic rats. Sprague Dawley male rats weighing 100$\pm$10g were randomly assigned to one normal and three STZ induced diabetic groups, which were subdivided into vitamin E free diet(DM 0E group), 40mg vitamin E per kg diet(DM 40E group) and 400mg vitamin E per kg diet(DM 400E group). Vitamin E level of normal group was 40 mg per kg diet. Diabetes was experimentally induced by intravenous injection of 55mg/kg of body weight of streptozotocin(STZ) in citrate buffer(pH 4.3) after 4 weeks feeding of experimental diets. Animals were sacrificed at the 6th day of diabetic states. There were no significant on body weights, food intakes, and food efficiency ratio before the diabetic occurrence. But after the injection of STZ, body weights and food efficiency ratios were significantly decreased and the food intakes was increased. Kidney weights were significantly increased in diabetic groups compared to normal group. However, there were no significant differences among the diabetic groups. Plasma insulin levels of diabetic groups were significantly decreased, whereas, blood sugar levels were increased compared to that of normal group. There were no significant differences among diabetic groups in plasma insulin and glucose levels. Activities of superoxide dismutase(SOD) in DM 0E and DM 40E groups were signi ficantly decreased by 33% and 27%, respectively, compared to normal group. But that of DM 400E group was increased by 35% compared to DM 0E group. Activity of glutathione peroxidase(GSHpx) in DM 0E group was decreased by 20% compared with normal group. GSHpx activity in DM 400E group was increased by 29% compared to normal group. The contents of vitamin E in kidney were 58% and 49% lower in DM 0E and DM 40E group, respectively, than normal group. There was no significant difference in renal vitamin E contents between DM-400E group and normal group. The contents of superoxide radical(O2 ) in kidney were 150% and 98%, respectively, higher in DM 0E and DM 40E groups than normalgroup. DM 400E and normal groups were similar levels in their superoxide radical contents of kidneys. These results indicate that vitamin E functioned as chain breaking antioxidant in kidney such as in other tissues.
Kim, Hae-Ja;Choi, Yun-Hee;Cho, Hwa-Eun;Hong, Hak-Gi;Han, Jung-Ho;Lee, Ki-Nam
Journal of Society of Preventive Korean Medicine
/
v.12
no.2
/
pp.13-25
/
2008
The purpose of this study was to investigate extract from mixed culture with Trichloloma matsutake mycelium in oriental medicine and cereal medium(OCM) to develop new material for pharmaceutical products and medicinal food for diabetes mellitus. To evaluate of hypoglycemic activity of OCM extracts, we examined the inhibitory activity of ${\alpha}$-glucosidasein OCM, blood glucose level and liver function of streptozotocin(STZ) induced diabetic rat. Experimental group was divided into 6 groups: first, it was divided into normal control group(hereafter NC group) and diabetes-induced group, and diabetes-induced group was subdivided into diabetic control group(DC group), treated by hot water extracts group(HE), ultra sonic waves, micro waves, and micro bubble extracts g roup(UE), crude polysaccharide of HE group (HEE) and crude polysaccharide of UE group(UEE) at a dose of 300mg/kg/body weight, respectively. In diabetic-induced groups, after streptozotocin was melted in 0.01M citrate buffer at 50mg/kg/body weight, when the non-fasting blood glucose levelwas 300 mg/dl or more in blood collected from the tail vein, it was regarded as diabetic induction and then such diabetic-induced experimental animals were used in this experiment. At the end of the experiment, blood glucose level increased by 4.19% in DC group but significantly decreased by 32.34%, 19.19%, 17.81% and 17.64%, respectively in UEEE, UE, HE, and HEE groups. In the cases of AST, ALT, and ALP, the experiment group treated with extracts showed significantly lowerblood glucose level than DC group. The levels of BUN and uric acid were found to be lower in the UMPM extract group(UE) than HW extract group(HE), which implies that herb medicine medium extracts in which Tricholoma matsutake mycelia were cultured are effective in reducing impaired liver function as well as high blood glucose level caused by diabetes. In addition, the administration of low temperature UMPM extracts was found to produce better results than that of high temperature hot water extracts. In this regard, it is expected that extracts from herb medicine obtained by cultivating Tricholoma matsutake mycelia will be widely used as new ingredients for foods and medicines for prevention and treatment of diabetes.
Cyclodextrin glucanotransferase (CGTase) from Paenibacillus sp. JB-13 was immobilized on various carriers by several immobilization methods such as ionic binding, covalent linkage and ultrafiltration to improve the process performance. The ultrafiltration and covalent linkage with CNBr-activated sepharose 4B were found as the best method for immobilization of CGTase. The ability of CGTase immobilization onto CNBr-activated sepharose 4B was as high as 18,000 units/g resin when the conditions was as follows: contact time 9 hrs at $37^{\circ}C$, pH 6.0, 100 nm and enzyme loading 24,000 units/g resin. The optimum conditions for production of 2-O-$\alpha$-D-Glucopyranosyl L-Ascorbic acid by immobilized CGTase turned out to be: pH 5.0, temperature $37^{\circ}C$, 20% substrate solution containing 8% (w/v) of soluble starch and 12% (w/v) of L-ascorbic acid sodium salt, 100 rpm, far 25 hrs and with 800 units of immobilized CGTase/ml substrate solution. Moreover the CGTase activity could be stably maintained for 8 times of repetitive reactions after removing products by ultrafiltration through YM 10 membrane.
Journal of the Korean Society of Food Science and Nutrition
/
v.31
no.3
/
pp.500-505
/
2002
Many studies have shown that hyperglycemia leads to an increase of lipid peroxidation in diabetic patients and animals, reflecting a rise reactive oxygen species production. It is increasingly recognized that brain is another site of diabetic organ damage. Accordingly, this study was to investigate the effect of dandelion on oxygen free radical generating and scavenging system of brain in streptozotocin (STZ)-induced diabetic rats. Male Wistar rats were divided into diabetic (control) and diabetic-dandelion supplemented groups. Dandelion was supplemented for 4 weeks with dandelion leaf and root powder (DLP, DRP) or dandelion leaf and root water extract (DLW, DRW) based on 11.4 g of raw dandelion/kg diet. Diabetes was induced by single injection STZ (55 mg/kg B.W., i.p.)in a citrate buffer. Oxygen free radical generating enzymes, cytochrome P-450, amino-pyrine N-demethylase, aniline hydroxylase and xanthine oxidase, were lowered in dandelion supplemented-groups compared to the control group. Superoxide dismutase, catalase and gluthathione peroxidase activities of brain were also lower in dandelion leaf and root supplemented-group than in the control group, whereas glutathione S-transferase activity and gluthathione content were increased in dandelion supplemented-groups compared to the control group. With regard to the lipid peroxidation products, the malondialdehyde content of brain was lower in dandelion supplemented groups. Therefore, it could be suggested that powder and water extract of dandelion leaf or root are beneficial in preventing diabetic complication from lipid peroxidation and free radical in brain of diabetic rat brain.
Two α-ʟ-arabinofuranosidases (BfdABF1 and BfdABF3) and a β-ᴅ-xylosidase (BfdXYL2) genes were cloned from Bifidobacterium dentium ATCC 27679, and functionally expressed in E. coli BL21(DE3). BfdABF1 showed the highest activity in 50 mM sodium acetate buffer at pH 5.0 and 25℃. This exo-enzyme could hydrolyze p-nitrophenyl arabinofuranoside, arabino-oligosaccharides (AOS), arabinoxylo-oligosaccharides (AXOS) such as 32-α-ʟ-arabinofuranosyl-xylobiose (A3X), and 23-α-ʟ-arabinofuranosyl-xylotriose (A2XX), whereas hardly hydrolyzed polymeric substrates such as debranched arabinan and arabinoxylans. BfdABF1 is a typical exo-ABF with the higher specific activity on the oligomeric substrates than the polymers. It prefers to α-(1,2)-ʟ-arabinofuranosidic linkages compared to α-(1,3)-linkages. Especially, BfdABF1 could slowly hydrolyze 23,33-di-α-ʟ-arabinofuranosyl-xylotriose (A2+3XX). Meanwhile, BfdABF3 showed the highest activity in sodium acetate at pH 6.0 and 50℃, and it has the exclusively high activities on AXOS such as A3X and A2XX. BfdABF3 mainly catalyzes the removal of ʟ-arabinose side chains from various AXOS. BfdXYL2 exhibited the highest activity in sodium citrate at pH 5.0 and 55℃, and it specifically hydrolyzed p-nitrophenyl xylopyranoside and xylo-oligosaccharides (XOS). Also, BfdXYL2 could slowly hydrolyze AOS and AXOS such as A3X. Based on the detailed hydrolytic modes of action of three exo-hydrolases (BfdABF1, BfdABF3, and BfdXYL2) from Bf. dentium, their probable roles in the hemiceullose-utilization system of Bf. dentium are proposed in the present study. These intracellular exo-hydrolases can synergistically produce ʟ-arabinose and ᴅ-xylose from various AOS, XOS, and AXOS.
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