The hypoglycemic effect of fractions of methanol extract of Cassia tora was investigated in diabetic rats. Sprague-Dawley male rats weighing 180∼230 g were divided into 6 groups. Diabetes was induced in the male rats by intravenous injection of streptozotocin at a dose of 45 mg/kg dissolved in citrate buffer (pH 4.5). The diabetic animals then had plasma glucose concentration of above 300 mg/㎗. Fractions of methanol extract of Cassia tora were administered orally into the diabetic rats for 14 days after streptozotocin injection. The food intake and body weight gain were monitored and plasma levels of glucose, triglyceride, free fatty acid, HDL-cholesterol, cholesterol and ALT activity were determined. Levels of glycogen, cholesterol and triglyceride in liver were analysed. Weights of liver and kidney were lighter in all the fraction-administered groups than streptozotocin-control group. Plasma glucose level was significantly decreased by the administration of butanol fraction at 14 days. Plasma triglyceride levels were lower in chloroform, butanol and H$_2$O fraction-administered groups compared to streptozotocin-control group. Plasma cholesterol and HDL-cholesterol levels were not significantly different in all groups. Administrations of each of the four fractions have decreased plasma free fatty acid level, ALT activity and liver triglyceride levels in the diabetic rats. Liver glycogen levels in chloroform, butanol and H$_2$O fraction-administered groups were higher than streptozotocin-control group. It is suggested from the results that butanol fraction of methanol extract of Cassia tora may contain the antihyperglycemic compounds.
${\beta}$-N-Acetyl-D-glucosaminidase B was highly purified with the following sequence of steps; DEAE-cellulose, CM-cellulose, and Sephadex G-200 gel filtration chromatograpies. The specific activity of the purified ${\beta}$ -N-acetyl-D-glucosaminidase B was 2.2 units/mg protein with 12.9 % yield and 196.2 fold purity. The purified ${\beta}$-N-acetyl-D-glucosaminidase B showed single band on polyacrylamide gel electrophoresis. The final preparation of ${\beta}$ -N-acetyl-D-glucosaminidase B was completely free friom arylsulfatase and ${\beta}$-glucuronidase. ${\beta}$ -N-Acetyl-D-glucosaminidase B had pH optimum of 4.5 in 0.5 M sodium citrate buffer. The molecular weight of ${\beta}$-N-acetyl-D-glucosaminidase B was 133,000 by Sephadex G-200 gel filtration. The Km value of ${\beta}$-N-acetyl-D-glucosaminidase B using p-nitrophenyl-N-acetyl-${\beta}$-D-glucosaminide as substrate was 1.0 mM and $V_{max}$ was 0.014 ${\mu}$ mole/min. ${\beta}$-N-Acetyl-D-glucosaminidase B was stable at $55^{circ}C$ for 70 minutes. The crude ${\beta}$ -N-acetyl-D-glucosamiinidase in 70 % ammonium sulfate retained 93 % activity after 7 months storage at -$55^{circ}C$. Bovine serum albumin, sodium chloride, and phosphate activated ${\beta}$ -N-Acetyl-D-glucosaminidase B. N-Acetyl-D-glucosamine, ${\alpha}$-methyl-D-mannoside, and acetate inhibited ${\beta}$ -N-acetyl-D-glucosaminidase B.
Park, Min-Young;Kim, Kyung-Hee;Jeong, Kyu-Shik;Kim, Hyeon-A
Journal of the Korean Society of Food Culture
/
v.22
no.1
/
pp.140-148
/
2007
Diabetic nephropathy has been increasing, although blood glucose and blood pressure can be controlled by angiotensin converting enzyme(ACE) or advanced glycosylation end products(AGE) inhibitors in the diabetic patients. We investigated the effect of dietary supplementation of sea tangle on the blood glucose, and pathological scoring of diabetic kidneys in the streptozotocin(STZ) induced diabetic rats. Male Sprague-Dawley rats were divided into normal rats fed control diet and diabetic rats fed control diet or control diet supplemented with powder or oater extract of sea tangle. Diabetes was induced by a single injection of STZ(60mg/kg, ip) in citrate buffer. The animals were fed the experimental diet and water for 13 weeks. Dietary supplementation of sea tangle decreased blood glucose in the diabetic rats. However, dietary supplementation of sea tangle did not affect the antioxidant enzyme activities, MDA content and pathology of diabetic kidneys. These results indicate that decreased blood glucose by sea tangle could not delay the progression of diabetic kidney disease.
This study was conducted to observe the physico-chemical exchange and effect of amino-carbonyl reaction between fructose and glycine . When various buffer solutions were added to equimolar mixture of fructose and glycine at pH 6.0 and $100^{\circ}C$, the browning effect was markedly observed by Mcllvaine buffer. Among the combinations of temperature and reaction time, the deep browning effect was obtained above $100^{\circ}C$, 3hr A marked browning effect obtained above pH 7.0 but little observed below pH 7.0. The browning effect was markedly increased at high fructose concentration. It required 4.0hrs and 32.9hrs to decrease 50% of initial concentration of fructose and glycine at $100^{\circ}C$ and pH 7 but 0.9hrs and 3.8hrs at $120^{\circ}C$, pH 7.0, respectively. The rate constant of fructose and glycine at $100^{\circ}C\;and\;120^{\circ}C$ were $1.78{\times}10^{-1},\;2.11{\times}10^{-2}\;and\;7.74{\times}10^{-1},\;1.83{\times}10^{-1}$, respectively. The formation of HMF was likely to follow the first order kinetics. The addition of 0.1M sodium sulfite, 0.1M sodium bisulfite and 0.1M calcium chloride to equimolar mixture (0.05M) surpressed the reaction up to 76.8%, 76.8% and 96.4%, respectively.
Present experiment was performed to establish the optimal reaction conditions for measurement of urinary gamma-glutamyltranspeptidase(${\gamma}$-GTP), N-acetyl-${\beta}$-D-glucosaminidase (AGS) and alanine aminopeptidase(AAP) activities in bovine and to investigate in vitro stability of the enzymes, within-run imprecision of the methods, and normal ranges. 1. The optimal wavelength for measurement of ${\gamma}$-GTP activity was 545nm. 2. The optimal pH of Tris-HCI buffer containing glycylglycine for measurement of urinary ${\gamma}$-GTP activity was 7.6~7.8(37$^{\circ}C$). 3. Coefficient of variance for within-run imprecision of urinary ${\gamma}$-GTP activity ranged from 4.8 to 7.2% and there was no significant difference among replications, 4. The optimal wavelength for measurement of urinary AGS activity was 405nm. 5. The optimal pH of citrate buffer for measurement urinary of AGS activity was 4.0(37$^{\circ}C$). 6. Coefficient of variance for within-run imprecision of urinary AGS activity ranged from 3.9 to 6.1% and there was no significant difference among replications. 7. The optimal wavelength for measurement of urinary AAP activity was 400nm. 8. The optimal pH of phosphate buffer for measurement of urinary AAP was 7.8. 9. Coefficient of variance for within-run imprecision of urinary AAP activity ranged from 2.5 to 4.8% and there was no significant difference among replications. 10. ${\gamma}$-GTP and AGS activities were increased significantly by gel-filtration. 11. Turbidity interfered with measurement of urinary AAP activity in bovine unless the specimen was gel-filterated. 12. Preservation of the specimen at 5$^{\circ}C$ or -20$^{\circ}C$ did not affect the AGS activity at least for 7 days after collection. 13. Preservation of the specimen at 5$^{\circ}C$ or 20$^{\circ}C$ did not affect the ${\gamma}$-GTP and AAP activities statistically, but some individual specimens revealed fluctuation during preservation. 14. ${\gamma}$-GTP, AGS and AAP activities revealed fluctuation by the tine of the day when the specimen was collected. 15. The normal ranges of urinary ${\gamma}$ -GTP, AGS and AAP activities were 6.60${\pm}$3.26(2.36-14.50), 1.31 ${\pm}$ 0.81(0.33-3.78), and 1.73 ${\pm}$ 0.55(0.77-3.03)U/l. respectively.
This experiment was performed to study the morphological responses of the pigment epithelial cell and the Bruch's membrane of the retina of rat following X-ray irradiation. Male rats were divided into normal and experimental groups. The heads of the rats, under sodium thiopental anesthesia, were exposed to 3,000 rads or 6,000 rads of radiation in a single dose, respectively. The source was a Mitsubishi Linear Accelerator ML-4MV. The target to skin distance was 80cm, and the. dose rate was 200 rads/min. The experimental groups were sacrificed on the 6th hour, 2nd and 6th day after X-ray irradiation. Under anesthesia, 1% glutaraldehyde-1% paraformaldehyde solution(0.1M Millonig's phosphate buffer, pH 7.3) was perfused through the left ventricle and ascending aorta. Pieces of the tissue taken from the posterior region of the retina were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde(0.1M Millonig's phosphate buffer, pH 7.3) and 1% osmium tetroxide(0.1M Millonig's phosphate buffer, pH7.3), and embedded in araldite mixture. The ultrathin sections contrasted with uranyl acetate and lead citrate were observed with JEM 100 CX-II electron microscope. The results were as follow; 1. The morphological changes of the pigment epithelial cells were not pronounced after exposure to 3,000 rads of X-ray. But on the 6th hour after exposure to 6,000 rads of X-ray, bulging nuclear membrane protruding into the cytoplasm and nuclear chromatin clumped into numerous masses along the nuclear membrane were observed. At the 2nd and 6th day post-irradiation, partial cytolysis or necrosis were seen. 2. The thickness of the Bruch's membrane of the experimental groups were increased in the time and dose range covered by this study, and splitting or diffusing basal laminae of the choriocapillary layer were observed frequently in the experimental group. Above results suggest that large amount(6,000 rads) of head irradiation induce direct hazardous effects on the pigment epitherial cells and Bruch's membrane of the retina of the rat, but pigment epithelial cells are more radioresistant than Bruch's membrane.
The authors observed the ultrastructure of the pigment cells of the frog, Rana nigromaculata Hallowell, during the hibernation. The specimens from the skin were fixed in 2.5% glutaraldehyde-paraform-aldehyde fixative in phosphate buffer at pH 7.2 prior to fixation in 2% osimium tetroxide, dehydrated in graded ethanol and acetone, embedded in Epon 812 mixture, and sectioned with LKB-ultramicrotome. the ultrathin sections were contrasted with uranyl acetate and lead citrate and observed with a JEOL-100B electron microscope. The results were as follows. In hibernating phase, pigment cells of the frog were consisted of the three kinds of chromatophores (xanthophore, iridophore and melanophore) in their dorsal skin. The traits of these cells were as follows. 1. Xanthophores A. Xanthophores were filled with pterinosomes and carotenoid vesicles. Many ribosomes, a few mitochondria and glycogen particles were dispersed in the cytoplasm. B. Pterinosomes were spherical or ellipsoidal in shape. They were divided into 6 types (type I, type II, type III, type IV, type V, type VI pterinosomes) by the their inner structure and especially, type I, type II, type III pterinosomes were well developed.
For the enzymatic production of $2-O-{\alpha}-D-glucopyranosyl-L-ascorbic$ acid (AA-2G) from ascorbic acid, rice seed was used as the source of ${\alpha}-glucosidase$ having transglucosylation activity. Among six rice varieties, cultivated in Korea, ${\alpha}-glucosidase$ activity of Oryza savita L. cv. Ilpumbyeo was the highest with 125.03 unit/ml and it had maximum specific activity with 8.52 unit/mg protein when rice seeds were grown for 3 days after germination. For the production of AA-2G using crude extract of O. savita L. cv. Ilpumbyeo, maltose was most effective glucose donor. The optimum concentration of maltose and ascorbic acid were 125 mM and 175 mM, respectively. The optimum concentration of ${\alpha}-glucosidase$ was 100 unit. The most effective buffer was 100 mM sodium citrate. The optimum pH and temperature were 5.0 and $60^{\circ}C$, respectively. Under the optimum condition, $108.43\;{\mu}M/unit$ of AA-2G was produced from ascorbic acid after 35 minutes of reaction, which corresponds to 6.2% of conversion ratio based on the amount of ascorbic acid used.
Kim, Han-Hwa;Noh, Yong-Tai;Chung, Young-Wha;Chi, Young-Duk
The Korean Journal of Zoology
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v.22
no.3
/
pp.103-114
/
1979
The authors observed the ultrastructure of the granular glands in the amphibian skin with an electron microscope. The specimens from the experimental animals (Bombina orientalis, Bufo bufo gargarizans, Rana nigromaculata and Rana rugosa) were fixed in 2.5% glutaraldehyde-paraformaldehyde fixative in phosphate buffer at pH 7.2 prior to fixation in 1% osmium tetroxide, dehydrated in graded ethanol and acetone, embedded in Epon 812 mixture, and sectioned with a LKB-ultramicrotome. the ultrathin sections were contrasted with uranyl acetate and lead citrate and observed with a JEOL-100B electron microscope. The results were as follws: 1. The granular gland in the amphibian skin consisted of the glandular epithelial and the myoepithelial cells. 2. The epithelial cells of the granular gland in the amphibian skin consisted of the dark cells but the light cells were also observed in that of Bombina orientalis. 3. The granular glands of the amphibian skin were in holocrine fashion. 4. The nuclei of the epithelial cells of the amphibian cutaneous granular glands were round or oval and showed small and large inforldings of nuclear envelope. Heterochromatins were mainly distributed near the nuclear envelope. Mitochondria were mainly distributed in the perinuclear portion and rough-surfaced endoplasmic reticulums were developed in the cytoplasm but smooth-surfaced endoplasmic reticulums were not well developed. 5. Secretory granules were round or oval and electron-dense and less electron-dense granules were observed. 6. The authors infer that the differences in electron density of the secretory granules in the granular glands of the amphibian skin are due to difference in the concentrations of secretory substances as related to the processes of its formation, and that those chemical components are identical.
Kim, Han-Hwa;Noh, Yong-Tai;Chung, Young-Wha;Chi, Young-Duk
The Korean Journal of Zoology
/
v.21
no.1
/
pp.29-39
/
1978
The authors observed the ultrastructure of the mucous glandular epithelial cells in the amphibian skin by mean of electron microscope. The specimens from the experimental animals were fixed in 2.5% glutaraldehyde-oaraformaldehyde fixative in phosphate buffer at pH 7.2 prior to fixation in 1% osmium tetroxide, dehydrated in graded ethanol and acetone, embedded in Epon 812 mixture, and sectioned with Sorvall MT-2 ultramicrotome. The ultrasections were contrasted with uranyl acetate and lead citrate and observed with a JEOL-100B electron microscope. The results were as follows: 1. The cutaneous mucous glands in amphibia consisted of the glandular epithelial and the myoepithelial cells. 2. Several different cells in ultrastructure were observed in the mucous glandular epithelium of the adult amphibian skin. a. The dark and the light cells were observed in Hynobius leechi. b. The mitochondria-rich and the round secretory granule-containing cells were observed in Bombina orientalis. c. The round secretory granule-containing and the foam-like granule mass-containing cells were observed in Kaloula tornieri. d. The cutaneous mucous gland of Rana nigromaculata were divided into two types: A and B-type glands. In the A-type mucous gland, the mitochondria-rich and the round secretory granule-containing cells and in the B-type mucous gland, the mitochondria-rich, the secretory granule-containing and the ER-rich cells were observed. 3. Based upon the above findings, the authors infer that the mucous granular epithelium of the amphibian skin consists of the mitochondria-rich undifferentiated, the secetory granule-containing and mature, and the ER-rich evacuated cells.
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