• Journal of Sericultural and Entomological Science
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• v.39 no.2
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• pp.119-133
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• 1997

Isozyme was used to characterize general protein patterns of genetic relationships among 303 silkworm stocks preserved in National Sericultural and Entomology Research Institute, RDA. Six isozymes (esterase, acid phosphatase, alkaline phosphatase, amylase, glucose-6-phosphate dehydrogenase and sucrase) from hemolymph, midgut, and digestive juice were employed to construct dendograms(UPGMA method) using a polycrylamide gel electrophoresis. A cluster analysis revealed four major group, which were divided into several subgroups within each group, contained assemglages of Japanese and Chinese races. Especially, genetic differentiation in the first and second group was greatest rather than within Japanese and Chinese races repectively and was concordant with the hypothesis of phyletic sorting of initial variability in China many years ago. Hypothesized recent introgression between groups was also plausible, but the eviednce suggested bidirectional gene flow between the Chinese and the Japnaese lineages. Interpreting the results in light of evidence from the current study, the genetic diversity and relationship showed in Korean silkworm race, Hansammyun reflected early and independent evolution from the Chinese ancestor, limited addition of new variability and phyletic sorting within Korean peninsula more than 4,000 years.

• Korean Journal of Clinical Laboratory Science
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• v.43 no.1
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• pp.6-11
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• 2011

In clinical chemistry tests, the interfering substances such as hemoglobin, lipid, bilirubin, and drugs, etc. can cause the changes of test results performed by spectrophotometrical methods. We evaluated the effects of interfering substances on the test results by adding interfering substances on the samples in the 19 kinds of clinical chemistry tests such as aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, lactate dehydrogenase, gamma-glutamyltransferase, total protein, albumin, glucose, total cholesterol, total bilirubin, triglyceride, uric acid, calcium, inorganic phosphours, high density lipoprotein cholesterol, low density lipoprotein cholesterol, creatinine, blood urea nitrogen, and C-reactive protein using newly implemented automatic chemical analyzer Toshiba TBA-C8000 under the direction of CLSI EP07-A guideline. Hemolytic samples show increased concentration of total protein, aspartate aminotransferase, alanine aminotransferase, and lactate dehydrogenase and reduced concentration of total bilirubin, alkaline phosphatase by interfering effect. Hyperlipemic samples show increased concentration of total protein and alkaline phosphatase and reduced concentration of low density lipoprotein cholesterol. The samples with conjugated bilirubinemia show increased concentration of inorganic phosphours, otherwise the samples with unconjugated bilirubinemia show no interference or allowable range in the test result.

• Animal Bioscience
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• v.36 no.11
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• pp.1693-1699
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• 2023

Objective: Cows that are nursing get around 80% of their glucose from liver gluconeogenesis. Propionate, a significant precursor of liver gluconeogenesis, can regulate the key genes involved in hepatic gluconeogenesis expression, but its precise effects on the activity of enzymes have not yet been fully elucidated. Therefore, the aim of this study was to investigate the effects of propionate on the activity, gene expression, and protein abundance of the key enzymes involved in the gluconeogenesis of dairy cow hepatocytes. Methods: The hepatocytes were cultured and treated with various concentrations of sodium propionate (0, 1.25, 2.50, 3.75, and 5.00 mM) for 12 h. Glucose content in the culture media was determined by an enzymatic coloring method. The activities of gluconeogenesis related enzymes were determined by enzyme linked immunosorbent assay kits, and the levels of gene expression and protein abundance of the enzymes were detected by real-time quantitative polymerase chain reaction and Western blot, respectively. Results: Propionate supplementation considerably increased the amount of glucose in the culture medium compared to the control (p<0.05); while there was no discernible difference among the various treatment concentrations (p>0.05). The activities of cytoplasmic phosphoenolpyruvate carboxylase (PEPCK1), mitochondrial phosphoenolpyruvate carboxylase (PEPCK2), pyruvate carboxylase (PC), and glucose-6-phosphatase (G6PC) were increased with the addition of 2.50 and 3.75 mM propionate; the gene expressions and protein abundances of PEPCK1, PEPCK2, PC, and G6PC were increased by 3.75 mM propionate addition. Conclusion: Propionate encouraged glucose synthesis in bovine hepatocytes, and 3.75 mM propionate directly increased the activities, gene expressions and protein abundances of PC, PEPCK1, PEPCK2, and G6PC in bovine hepatocytes, providing a theoretical basis of propionate-regulating gluconeogenesis in bovine hepatocytes.

• Journal of Food Hygiene and Safety
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• v.8 no.4
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• pp.195-204
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• 1993

Basolateral and brush border membrane (BLM and BBM) vesicles of renal proximal tubules were prepared from adult male New Zealand White rabbits to evaluate as a potential model for assessment of nephrotoxicity. PAH uptakes using BLMV, glucose and leucine uptakes using BBMV were measured in the rabbits treated cephaloridine. In addition, urinalysis and histopathological studies were performed to investigate the correlationship with membrane vesicle uptakes. The results were as follows: (1) the activite of Na+, K+ -ATPase was enriched 12.3-fold in vasolateral memvrane vesicles (BLMV) and the specific activity of alkaline phosphatase in purified brush border memvrane vesicles (BBMV) was enriched 10.1-fold compared with each of microsomal homogenate. (2) In the uptake experiments, cephaloridine decreased initial and probenecid-sensitive PAH uptakes in BLMV. (3) Cephaloridine tested decreased initial and phlorizin-sensitive glucose uptakes in BBMV. (4) Cephaloridine tested decreased initial and Na+-dependent leucine uptakes in BBMV. (5) Cephaloridine tested significantly increased the urinary excretion of glucose and activity of ${\gamma}$-GTP. (6) Cephaloridine tested caused moderate necrotic changes in renal tubular cells and formation of urinary cast in the lumina of Henle's loop and collecting tubules besides the swelling of renal tubules.

• BMB Reports
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• v.31 no.5
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• pp.519-523
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• 1998

We have previously reported that anti-glucose-6-phosphate dehydrogenase (G6PD) serum raised against native G6PD (nG6PD) enzyme recognized nG6PD antigen poorly in competitive enzyme-linked immunosorbent assay (ELISA) (Kim, 1997). In the present study, we investigated whether anti-G6PD serum raised against nG6PD can react with denatured G6PD effectively in competitive ELISA. We used partially active G6PD (paG6PD) by repeated freeze-thawing or SDS-denatured G6PD (SDS-G6PD) as both immobilized and soluble antigens, and anti-G6PD serum raised against nG6PD for competitive ELISA. The polystyrene cuvettes coated with either paG6PD or SDS-G6PD were challenged with a mixture of a limiting amount of anti-G6PD serum and various doses of paG6PD or SDS-G6PD as competitors, followed by incubation with alkaline phosphatase-anti-IgG conjugate. The competitive ELISA with paG6PD or SDS-G6PD antigen exhibited the sigmoidal dose-response curve characteristic of competition immunoassays. Furthermore, Triton-denatured G6PD (Triton-G6PD) was used in competitive ELISA. The paG6PD, SDS-G6PD, or Triton-G6PD used as competitors increased the inhibition of antibody binding to immobilized either of nG6PD or denatured G6PD compared with nG6PD competitor. The inhibition by denatured G6PD competitors was more pronounced at high competitor concentrations than at low counterparts. We conclude that anti-G6PD serum raised against nG6PD can effectively react with denatured G6PD in competitive ELISA and that our anti-G6PD serum recognizes denatured enzymes better than active enzymes.

• Journal of Ginseng Research
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• v.47 no.3
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• pp.420-428
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• 2023

Background: Ginsenoside F2 (GF2), a minor component of Panax ginseng, has been reported to possess a wide variety of pharmacological activities. However, its effects on glucose metabolism have not yet been reported. Here, we investigated the underlying signaling pathways involved in its effects on hepatic glucose. Methods: HepG2 cells were used to establish insulin-resistant (IR) model and treated with GF2. Cell viability and glucose uptake-related genes were also examined by real-time PCR and immunoblots. Results: Cell viability assays showed that GF2 up to 50 μM did not affect normal and IR-HepG2 cell viability. GF2 reduced oxidative stress by inhibiting phosphorylation of the mitogen-activated protein kinases (MAPK) signaling components such as c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase 1/2 (ERK1/2), and p38 MAPK, and reducing the nuclear translocation of NF-κB. Furthermore, GF2 activated PI3K/AKT signaling, upregulated the levels of glucose transporter 2 (GLUT-2) and GLUT-4 in IR-HepG2 cells, and promoted glucose absorption. At the same time, GF2 reduced phosphoenolpyruvate carboxykinase and glucose-6-phosphatase expression as well as inhibiting gluconeogenesis. Conclusion: Overall, GF2 improved glucose metabolism disorders by reducing cellular oxidative stress in IR-HepG2 cells via MAPK signaling, participating in the PI3K/AKT/GSK-3β signaling pathway, promoting glycogen synthesis, and inhibiting gluconeogenesis.

• BMB Reports
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• v.30 no.5
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• pp.326-331
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• 1997

To construct a competitive ELISA standard curve for the detection of glucose-6-phosphate debydrogenase (G6PD), we used highly purified native G6PD (nG6PD) as both immobilized and soluble antigens and anti-G6PD serum raised against nG6PD as antibody. The polystyrene cuvettes coated with nG6PD were challenged with a mixture of a limiting amount of anti-G6PD serum and various doses of nG6PD as competitors followed by incubation with alkaline phosphatase-anti-IgG conjugate. The competitive ELISA did not exhibit the typical sigmoidal dose-response curve characteristic of competition immunoassays under the optimal concentrations of antigen and antibody. The soluble nG6PD used as competitor failed to effectively inhibit the binding of antibodies to the immobilized nG6PD. The addition of NADP, a cofactor of G6PD enzyme, to coating buffer used for immobilizing nG6PD to the cuvettes and PBS-Tween-BSA buffer for diluting competitors did not improve the inhibition of antibody binding to immobilized nG6PD by soluble n/G6PD. The addition of BSA to coating buffer did not increase inhibition, either. Surprisingly, when partially active G6PD (paG6PD), obtained by repeated freeze-thawing, was used as competitor, the antibody binding to either immobilized nG6PD or immobilized paG6PD was inhibited 49-58%. We conclude that an effective competitive ELISA system with nG6PD enzyme and anti-G6PD serum for the detection of G6PD may not be established due to the poor inhibition of antibody binding to immobilized nG6PD by soluble nG6PD under the present assay conditions and that the inhibition may be improved by using an inactivated enzyme as competitor regardless of the type of immobilized antigen used. These results imply that the immobilized nG6PD may undergo denaturation upon binding to the polystyrene cuvettes and that our anti-G6PD serum may recognize denatured enzyme better than active enzyme.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.9
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• pp.1330-1335
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• 2005

Red brownish p-pheylenediamine (PPD) has been widely used hair dye for women. The dye was known to cause systemic anaphylaxis, dermatitis and bladder cancer. But the effect of PPD toxicity with oxygen free radical has not been studied. This study investigated the degree of skin injury by PPD. PPD ($2.5\%$ PPD in $2\%\;NH_{4}OH$) was applied to the rat skin ($25 mg/16.5\;cm^2$) 3 or 5 times every other day. Histopathological findings demonstrated the proliferation of epithelial cells and the increased keratinization by PPD. The activities of glucose 6-phosphatase (G6Pase) was decreased and acid phosphatase (ACP) was increased in PPD-applied rat skin. Groups in which PPD was applied 5 times were more damaged than groups applied 3 times. To examine the relationship between tissue damage and oxygen free radicals, effect of PPD on xanthine oxidase (XO) activity was measured and XO activity was more significantly increased in the group treated with PPD 5 times than 3 times. However, reduced glutathione (GSH) content, and the activities of catalase (CAT), super-oxide dismutase (SOD) and glutathione S -transferase (GST) were more decreased in PPD-applied groups than in controls. Even though the activities of XOD was not changed in the group treated with PPD 3 times, the decreased activities of oxygen free radical system and the damaged skin tissue were observed. This result might be caused by the production of toxic PPD metabolites in rat skin. In conclusion, topical PPD application led to skin injury in a dose-dependent manner, probably due to the generation rate of oxygen free radical.

• Korean Journal of Veterinary Research
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• v.34 no.3
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• pp.619-626
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• 1994

The purpose of present study was to examine pharmacological effect of Artemisia Iwayomogi methanol extract(AIME) on biochemical parameters(activities of AST, ALT, LDH, and ALP, contents of total bilirubin, total protein, albumin and A/G ratio in serum and levels of hepatic microsomal lipid peroxide and glucose-6-phosphatase activities) against hepatic injury by carbon tetrachloride($CCl_4$) in rats. Increased AST, ALT and LDH activities by $CCl_4$ were decreased in AIMS treatment group at 48 or 72 hours. Together, increased ALP activity by $CCl_4$ almost returned toward normal value in AIME treatment group at 72 hours. Serum total bilirubin contents increased to 87, 79 and 31% compared with normal group by $CCl_4$ which were decreased to 64, 42 and 26% in AIME treatment group at 24, 48 and 72 hours, respectively. Decreased contents of total protein and albumin, and A/G ratio by $CCl_4$ were recovered in AIME treatment group. Hepatic microsomal lipid peroxide levels(nmol malonic dialdehyde/100mg protein) increased to 140, 95 and 78% compared with normal group by $CCl_4$ which were decreased to 107, 74 and 65% in AIME treatment group at 24, 48 and 72 hours, separately. Hepatic microsomal glucose-6-phosphatase activities decreased to 60, 50 and 53% compared with normal group by $CCl_4$ at 24, 48 and 72 hours, respectively, which were increased at 72 hours in AIME treatment group. In conclusion, AIME enhanced the amelioration process from $CCl_4$-induced lipid peroxidation, degeneration of liver cell, and impairment of protein and bilirubin metabolisms.

• Journal of Ginseng Research
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• v.13 no.1
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• pp.92-97
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• 1989

This study investigated the effects of high light intensity (100 KLw) and high temperature (45 ℃, dark) on enzyme (glucose-6-phosphate dehydrogenase, acid phosphatase, catalase, peroxidase, and proteinase) activities and characteristics of Panax ginseng C.A. Meyer leaves. Enzyme activity and protein content decreased rapidly under treatment with high light intensity In P ginseng the thermal stabilities of catalase and peroxidase were high (above 70%), and the coagulation rates of soluble proteins were low (below 17%). Therefore, the decrease in enzyme activity and protein content was not caused by increase in leaf temperature due to the high light intensity, but by increase in proteolytic activities. The photochemical formation rate of superoxide radical (O-2) was higher in the P ginseng leaf extracts than in Solanum nigmm, and was accelerated by addition of crude saponin to the buffer extracts.