• Journal of Life Science
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• v.19 no.9
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• pp.1218-1225
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• 2009

Difractose anhydrides (DFAs) is studied as a sweetener for diabetics because of its structural property. DFAs have four types: DFA I, III, IV (degradation of levan) and V (degradation of inulin). Especially, DFA IV has been shown to enhance the absorption of calcium in experiments using rats. Levan fructotransferase is an enzyme for producing di-d-fructose-2,6':6,2-dianhydride (DFA IV). To identify structural characterization, we purified wild-type and mutants (D63A, D195N and N85S) of levan fructotransferase (LFTase) from Microbacterium sp. AL-210. These proteins were purified to apparent homogeneity by Ni-NTA affinity column, Q-sepharose ion exchange and gel filtration chromatography and detected by SDS-PAGE. They were also analyzed by circular dichroism (CD) measurements, JNET secondary structure prediction, activity measurements at various temperatures, and pH analysis. The optimum pH for the enzyme-catalyzed reaction was pH 7.5 and optimum temperature was observed at $55^{\circ}C$. Along with wild-type LFTase, mutants were analyzed by CD measurement, fluorescence analysis and differential scanning calorimetry (DSC). N85S showed less $\alpha$-helix and more $\beta$ strand than others. Also, N85S showed almost the same curve as wild-type in their steady-state fluorescence spectra, whereas mutant D63A and D195N showed higher intensity than wild-type. The amino acid sequence of wild-type LFTase was compared to the sequences of exo-inulinase from Aspergillus awamori, a plant fructan 1-exohydrolase from Cichorium intybus, and Thermotogo maritime (Tm) invertase and showed a high identity with Exo-inulinase from Aspergillus awamori.

• Childhood Kidney Diseases
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• v.15 no.2
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• pp.138-145
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• 2011

Purpose : To test whether the expression of ${\beta}$-catenin, a component of podocyte as a filtration molecule, would be altered by puromycin aminonucleoside (PAN) in the cultured podocyte in vitro. Methods : We cultured rat glomerular epithelial cells (GEpC) with various concentrations of PAN and examined the distribution of ${\beta}$-catenin by confocal microscope and measured the change of ${\beta}$-catenin expression by Western blotting and reverse transcriptase-polymerase chain reaction (RT-PCR). Results :We found that ${\beta}$-catenin relocalized from peripheral cytoplasm to inner cytoplasm, therefore, intercellular separations were seen in confluently cultured cells by high concentrations of PAN in immunofluorescence views. In Western blotting of GEpC, PAN ($50{\mu}g/mL$) decreased ${\beta}$-catenin expression by 34.9% at 24 hrs and 34.3% at 48 hrs, compared to those in without PAN condition (P<0.05). In RT-PCR, high concentrations ($50{\mu}g/mL$) of PAN also decreased ${\beta}$-catenin mRNA expression similar to protein suppression by 25.4% at 24 hrs and 51.8% at 48 hrs (P<0.05). Conclusion : Exposure of podocytes to PAN in vitro relocates ${\beta}$-catenin internally and reduces ${\beta}$-catenin mRNA and protein expression, which could explain the development of proteinuria in experimental PAN-induced nephropathy.

• Korean Journal of Pharmacognosy
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• v.40 no.3
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• pp.196-204
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• 2009

This study was carried out to investigate the in vivo and in vitro inhibitory effect of a traditional herbal complex (HC) extract prepared from a mixture of four oriental herbs (Dioscorea Rhizoma, Glycine soja Sieb. et Zucc, Bombycis corpus, Fermented Glycine soja) that have been widely used for the treatment and prevention of diabetes mellitus on hyperglycemia. The water extract of HC showed potent inhibitory effect on $\alpha$-glucosidase with $IC_{50}$ value of 1.24 mg/mL. Additionally, the ethanol extract of HC was also found to exhibit significant inhibitory effect against protein tyrosine phosphatase $1{\beta}$ ($PTP1{\beta}$), which is known as a major regulator of both insulin and leptin signaling. In the $PTP1{\beta}$ inhibitory assay, the most active n-hexane fraction obtained from the ethanol extract of HC, was identified as a mixture of fatty acid derivatives by gas chromatography-mass spectrometry (GC-MS). In high-fat diet-low dose streptozotocin (STZ)-induced diabetic rat, the water extract of HC improved the oral glucose intolerance as compared with rosiglitazone. HC also caused a marked decrease of body weight and fasting blood glucose and a significant improvement on glucose tolerance in metabolic syndrome mice model. These findings support that this traditional HC may be useful in the control of blood glucose in diabetes mellitus and metabolic syndrome.

• Nuclear Medicine and Molecular Imaging
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• v.43 no.1
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• pp.72-78
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• 2009

Purpose: Bone mineral density (BMD) measurements need to be precise enough to be capable of detecting small changes in bone mass of rats. Using a regular dual-energy X-ray absorptiometry (DXA), we measured many BMD of various skeletal sites in rats to examine precision of DXA in relation to the repositioning on the bones of rats. Materials and Methods: Using DXA and small animal software, scans were performed 4 times in all 12 male rats without repositioning (Group 1a). Another four scans for 6 of 12 rats were done with repositioning between scans (Group 2). Customized regions of interest (ROIs), encapsulate the right hind limb, L1-4, skull and pelvic bones were drawn at each measurement. The precision of the measurements was evaluated by measuring the coefficient of variation (CV) of four measurements of BMD at each skeletal site of all rats with or without repositioning. Significance of differences between group 1b (six rats out of group 1a, which were come under group 2) and group2 were evaluated with Wilcoxon Signed Rank Sum Test. Results: CVs obtained at different skeletal sites of all measurements in Group 1b and 2. It was $3.51{\pm}1.20$, $ 2.62{\pm}1.20$ for the hindlimb (p=0.173), $3.83{\pm}2.02$, $4.59{\pm}2.02$ for L1-4 (p=0.600), $3.73{\pm}1.87$, $1.53{\pm}0.89$ for skull (p=0.046), and $2.92{\pm}0.60$, $1.45{\pm}0.60$ for pelvic bones (p=0.075). Conclusion: Our study demonstrates that the DXA technique has the precision necessary when used to assess BMD for various skeletal sites in rats regardless of repositioning.

• Journal of Life Science
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• v.18 no.11
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• pp.1584-1591
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• 2008

The effect of Chlorella hot water extract (CE) on hyperglycemia in streptozotocin- induced diabetic rats has not been studied. Therefore, hypoglycemic effect of CE in type I streptozotocin- induced diabetic rats was studied. Rats were fed a semisynthetic diet supplemented with either 3% (the STZ+CE3) and 6% (the STZ+CE6) CE or no supplement the Normal and the STZ-Control rats for 4 weeks. The concentrations of fasting and non-fasting blood glucose were higher in the STZ-Control rats than in the Normal rats, but this rise was lowered in the STZ+CE3 and the STZ+CE6 rats. Serum insulin concentrations were decreased with STZ injection, however, the decreased levels were almost restored to the Normal level with CE supplementation. The increased serum fructosamine levels associated with hyperglycemia were decreased with the CE treatment. The morphology of pancreatic islets in the Normal rat was round and maintained a typical arrangement. The STZ-Control pancreatic beta-cells were found to have significant swelling and severely morphological damaged, however, pancreatic tissue damage by STZ in the CE-supplemented diet group was ameliorated. This study shows that Chlorella hot water extract had a hypoglycemic effect on the STZ-diabetic rats via either increased insulin secretion during recovery or the prevention of STZ-induced pancreatic damage.

• Journal of Life Science
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• v.22 no.5
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• pp.634-641
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• 2012

It was reported that the novel compounds (LP9M80-H) of $Liriope$ $platyphylla$ regulate glucose transporter (Glut) biosynthesis by activating the insulin-signaling pathway in the liver and brain of ICR mice. To investigate the therapeutic effects of LP9M80-H on the pathology of diabetes and obesity, alterations of key factors related to symptoms were analyzed in the Otsuka Long Evans Tokushima Fatty (OLETF) rats treated with LP9M80-H for 2 weeks. The abdominal fat masses in the LP9M80-H-treated group were lower than the vehicle-treated group, although there was no difference in body weight between the two groups. Additionally, when compared to the vehicle-treated group, LP9M80-H treatment induced a significant decrease in glucose levels and an increase in the insulin concentration in the blood of OLETF rats. A high level of insulin protein was also detected in pancreatic ${\beta}$ cells of LP9M80-H-treated OLETF rats. A significant reduction in the concentration of lipids and adiponectin was detected only in LP9M80-H-treated OLETF rats. Furthermore, the expression of insulin receptor ${\beta}$ and the insulin receptor substrate (IRS) was dramatically decreased in LP9M80-H-treated OLETF rats compared to the vehicle-treated group. Of the glucose transporters located downstream of the insulin-signaling pathway, glucose transporters (Glut) -2 and -3 were significantly decreased in LP9M80-H-treated OLETF rats, while the level of Glut-4 was maintained under all conditions. Therefore, these results suggest that LP9M80-H may contribute to relieving symptoms of diabetes and obesity through glucose homeostasis and regulation of lipid concentration.

• Journal of Life Science
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• v.19 no.10
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• pp.1417-1423
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• 2009

Previous studies have suggested that docosahexaenoic acid (DHA) supplementation into n-3 fatty acid deficient diet improved spatial learning performance, but there was no significant difference in brain related function when DHA was added into a n-3 fatty acid adequate diet. Here, we investigated the effect of adding DHA into an n-3 fatty acid deficient or adequate diet on brain and liver fatty acid composition. On the second day after conception, Sprague Dawley strain dams were divided into four groups as follows; n-3 fatty acid deficient (Def), n-3 fatty acid deficient plus DHA (Def+DHA, 10.2% DHA), n-3 fatty acid adequate (Adq, 3.4% linolenic acid), and n-3 fatty acid adequate plus DHA (Adq+DHA, 3.31% linolenic acid plus 9.65% DHA). After weaning, male pups were fed on the same diets of their respective dams until adulthood. In brain fatty acid composition, the Def group showed a lower brain DHA (64% decrease), which was largely compensated for by an increase in docosapentaenoic acid (22:5n-6). Brain DHA in the Def+DHA group was increased to almost the same extent as in the Adq and Adq+DHA groups and there were no significant differences among them. Liver fatty acid composition showed a similar pattern to that of the brain, but liver DHA in the Def+DHA showed the highest percentage among the diet groups. In conclusion, n-3 fatty acid deficiency from gestation to adulthood leads to decreased brain DHA, which has been shown to be highly associated with poor spatial leaning performance. Thus, adequate brain DHA levels are required for optimal nervous function.

• The Korean Journal of Food And Nutrition
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• v.29 no.4
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• pp.521-528
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• 2016

This study was conducted to investigate changes in the proximate composition, antioxidant activities, and ${\alpha}$-glucosidase inhibitory activity of Opuntia ficus-indica (OFI) cladodes cultivated in Jeju (JJ1, JJ2, JJ3) and Jeonnam (JN1, JN2). The difference in the proximate composition (crude protein, lipid and ash content) of OFI between the two regions was not significant. Ca, Mg and Na were the major mineral components of OFI. The ascorbic acid content of OFI ranged from 57.87 to 143.72 mg/100 g. A 70% ethanol extract was used to investigate the antioxidant content and activity as well as the ${\alpha}$-glucosidase inhibitory activity. The total polyphenol and flavonoid contents of OFI were 38.69~55.29 and 3.33~4.03 mg/g, respectively. The antioxidant activities based on the DPPH and ABTS free radical scavenging assays were 45.19~61.52% and 39.15~51.96%, respectively, at a concentration of 1 mg/mL. The inhibitory activity of OFI extracts against rat intestinal ${\alpha}$-glucosidase was 29.72~45.73% at 1 mg/mL concentration, and JN1 showed the highest ${\alpha}$-glucosidase inhibitory activity. This information could be very useful for authentication of Opuntia species with the highest potential as sources of nutritional and therapeutic elements.

• The Korean Journal of Food And Nutrition
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• v.22 no.1
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• pp.48-56
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• 2009

This study was conducted to examine the effects of Opuntia ficus-indica complexes(OCB) on the intake of water and food, body weight, blood glucose levels, glucose tolerance and lipid metabolism in streptozotocin(STZ)-induced diabetic rats. Thirty-two male Sprague-Dawley rats were assigned to four different groups; non-diabetic control(NC), diabetic control (DC), diabetic OCB of 2%(OCB-2), and diabetic OCB of 5%(OCB-5). The animals were fed on each experimental diet for 3 weeks. The DC, OCB-2 and OCB-5 groups showed a higher intake of water and food than the NC group. The fasting blood glucose levels were 100 $ mg/d{\ell}$ and 416 $ mg/d{\ell}$ for the NC and DC groups, respectively. The OCB-5 group presented a significantly low fasting glucose level of 21%(P<0.05), while OCB-2 group had a decrease of 13% compared to the DC group. As for the results of the glucose tolerance test, the highest blood glucose level was observed for all the groups at 30 minutes after the glucose injections as well as higher plasma insulin levels in the OCB-5 group. Plasma total cholesterol, triglyceride, non-esterified fatty acids(NEFA) and LDL-cholesterol concentrations were also lower in the OCB-2 and OCB-5 groups. The experimental diet did not affect the HDL-cholesterol levels. The overall results suggest that the higher intake of food by the OCB-2 and OCB-5 groups improved the blood glucose levels and lipid metabolism in STZ-induced diabetic rats.

• Journal of Yeungnam Medical Science
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• v.10 no.2
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• pp.313-337
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• 1993

The aim of this study was to clarify the role of Kupffer cells in the mechanism of endotoxin-induced liver injury. The study on fine structure of Kupffer cells was performed after the injection of endotoxin. The endotoxin(Escherichia coli lipopolysaccharide 026 : B6. 1.5mg/100 g of body weight) was intraperitoneally injected in Sprague-Dewley rats. Animals were sacrificed at 1/4, 1/2, 1, 2, 4, 8, 16, 24, 72 and 120 hours after the injection of endotoxin. Livers were extirpated and processed to be examined by light and electron microscopy. The results obtained were summerized as follows: Early changes observed in liver after endotoxin injection included the increased number and hypertrophy of Kupffer cells, infiltration of neutrophils and presence of fibrin thrombi within the sinusoids. The continuous increase of the Kupffer cells in number with hypertrophy, congestion and infiltration of inflammatory cells within the sinusoids were observed. Hepatocytes showed fatty change and occasional necrosis. At 72 hours the congestion decreased. At 120 hours the number of Kupffer cells was increased, but the morphology of Kupffer cells became similar to that of the control group. The numbers and sizes of primary and secondary lysosomes and amount of euchromatin of Kupffer cells increased. Swellings and increase in number of mitochondria, Golgi complex, smooth endoplasmic reticulum, rough endoplasmic reticulum were evident. Microthrombi were present within the sinusoids. The swelling of rough endoplasmic reticulum and mitochondria, decrease of glycogen particles, fatty change, hypoxic vacuoles, pyknotic nuclei and occasional necrosis were observed in hepatocytes. At 72 hours the number of secondary lysosomes in Kupffer cells decreased. At 120 hours the morphology of Kupffer cells became similar to that of the control group. According to these results, it was postulated that the endotoxin was initially taken up by pinocytosis into Kupffer cells and degraded in secondary lysosomes of activated Kupffer cells. Kupffer cells may play an important role in the defense mechanism of liver during endotoxemia. The dysfunction of Kupffer cells and ischemia by sinusoidal microthrombi may cause liver injury.