• The Journal of Internal Korean Medicine
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• v.32 no.4
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• pp.550-564
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• 2011

Objectives : Oxidative stress seems to play a major role in mechanisms by which ethanol causes liver injury. Previous studies have shown that treatment with Yinjinchunggan-tang (Yinchenqinggan-tang, YJCGT) has protective effects on alcoholic liver disease. The aim of this study was to investigate the protective effects of YJCGT on alcohol-induced oxidative stress. Materials and Methods : In vitro, we evaluated the inhibitory activities of YJCHT on DPPH(1,1-diphenyl-2-picryl-hydrazyl), xanthine oxidase, trypsin, and hyaluronidase. In a cell culture model, we measured cell viability and proliferation, and the activities of superoxide dismutase (SOD), and catalase (CAT) after YJCGT treatment in C34 and E47 cell lines, and HepG2 cells transfected with/ without cytochrome P450IIE1 (CYP2E1) gene. In vivo, we estimated serum level of hepatic biochemical markers, and alcohol concentration in the blood. Results : YJCGT showed significant free radical scavenging activity against DPPH and xanthine oxidase and decreased hyaluronidase activity effectively in vitro. YJCGT also increased cell viability, and proliferation in C34 and in E47 cell lines, and increased activities of superoxide dismutase, and catalase in C34 and in E47 cell lines. YJCGT reduced serum AST, LDH, and total cholesterol level in some of the results, and reduced blood alcohol concentration in vivo, as well. Conclusions : This study suggests that YJCGT has protective effects on oxidative stress by inhibiting alcohol-induced suppression of antioxidant enzyme activities.

• Journal of Microbiology and Biotechnology
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• v.21 no.6
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• pp.617-626
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• 2011

Transglutaminase from Streptomyces mobaraensis is an enzyme of unknown function that cross-links proteins to high molecular weight aggregates. Previously, we characterized two intrinsic transglutaminase substrates with inactivating activities against subtilisin and dispase. This report now describes a novel substrate that inhibits papain, bromelain, and trypsin. Papain was the most sensitive protease; thus, the protein was designated Streptomyces papain inhibitor (SPI). To avoid transglutaminase-mediated glutamine deamidation during culture, SPI was produced by Streptomyces mobaraensis at various growth temperatures. The best results were achieved by culturing for 30-50 h at $42^{\circ}C$, which yielded high SPI concentrations and negligibly small amounts of mature transglutaminase. Transglutaminasespecific biotinylation displayed largely unmodified glutamine and lysine residues. In contrast, purified SPI from the $28^{\circ}C$ culture lost the potential to be cross-linked, but exhibited higher inhibitory activity as indicated by a significantly lower $K_i$ (60 nM vs. 140 nM). Despite similarities in molecular mass (12 kDa) and high thermostability, SPI exhibits clear differences in comparison with all members of the wellknown family of Streptomyces subtilisin inhibitors. The neutral protein (pI of 7.3) shares sequence homology with a putative protein from Streptomyces lavendulae, whose conformation is most likely stabilized by two disulfide bridges. However, cysteine residues are not localized in the typical regions of subtilisin inhibitors. SPI and the formerly characterized dispase-inactivating substrate are unique proteins of distinct Streptomycetes such as Streptomyces mobaraensis. Along with the subtilisin inhibitory protein, they could play a crucial role in the defense of vulnerable protein layers that are solidified by transglutaminase.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.12
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• pp.1708-1714
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• 2005

Keratinocyte growth factor (KGF) functions in epithelial growth and differentiation in many tissues and organs. KGF is expressed in the uterine endometrial epithelial cells during the estrous cycle and pregnancy in pigs, and receptors for KGF (KGFR) are expressed by conceptus trophectoderm and endometrial epithelia. KGF has been shown to stimulate the proliferation and differentiation of conceptus trophectoderm. However, the role of KGF on the endometrial epithelial cells has not been determined. Therefore, this study determined the effect of KGF on proliferation and differentiation of endometrial epithelial cells in vitro and in vivo using an immortalized porcine luminal epithelial (pLE) cell line and KGF infusion into the uterine lumen of pigs between Days 9 and 12 of estrous cycle. Results showed that KGF did not stimulate proliferation of uterine endometrial epithelial cells in vitro and in vivo determined by the $^3$H]thymidine incorporation assay and the proliferating cell nuclear antigen staining, respectively. Effects of KGF on expression of several markers for epithelial cell differentiation, including integrin receptor subunits $\alpha$4, $\alpha$5 and $\beta$1, plasmin/trypsin inhibitor, uteroferrin and retinol-binding protein were determined by RT-PCR, Northern and slot blot analyses, and immunohistochemisty, and KGF did not affect epithelial cell differentiation in vitro and in vivo. These results show that KGF does not induce epithelial cell proliferation and differentiation, suggesting that KGF produced by endometrial epithelial cells acts on conceptus trophectoderm in a paracrine manner rather than on endometrial epithelial cells in an autocrine manner.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.10
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• pp.1447-1451
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• 2004

An experiment was conducted to determine the effects of different dietary crude protein (CP) levels on growth performance, nutrient utilization, small intestine protease activity and immunity index of weaner to 2 month-old New Zealand rabbits. Eighty weaner rabbits were allocated in individual cages to five treatments in which they were fed diets with CP at 14%, 16%, 18%, 20% and 22%, respectively. The growth performance and nutrient digestibility of rabbits increased firstly when dietary CP increased, then decreased. The average daily gain was the highest and feed conversion rate was the lowest when dietary CP reached 20%, namely 34.9 g/d and 2.74:1, respectively. Maximum CP digestibility was 72.1% in the 18% CP group, maximum crude fiber digestibility of 28.4% occurred in the 16% CP group and was significantly different from other treatments (p<0.01), apparent digestibility of Lys and Val followed the same trend as CP digestibility, and reached their maximum when dietary CP was 18%. Apparent digestibility of Cys, Tyr, Leu and Thr also had a similar trend to CP digestibility. Nitrogen retention (RN) increased with CP level (p>0.05), and was highest for 20% CP treatment (1.5 g/d). The effect of CP level on the rate of digestible nitrogen (DN) converted RN was small. The spleen index, thymus index, chymotrypsin and trypsin activities in small intestine were highest when dietary CP was 16%, which were 1.0, 2.8, 15.7 U/g and 125.7 U/g, respectively. There was no significant difference among treatments (p>0.05). According to the above results, the appropriate dietary CP level from weaner to 2 month-old meat rabbits was 18-20%.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.6
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• pp.899-904
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• 2018

Objective: This experiment studied the effects of first feed intake time post-hatch on growth performance, nutrient apparent metabolic rate and intestinal digestive enzyme activities in broilers. Methods: Two thousand five hundred and twenty LingNan Yellow broilers were randomly allotted to seven treatments with six replicates of 60 each. The only experimental factor was the first feed intake time which was 18, 24, 30, 36, 42, 48, and 54 hours after hatching. The whole experiment lasted for 21 days. Results: During the whole period, the 30 h treatment had the best body weight and average daily gain (p<0.05), followed by the 24 h group performance optimization. Also, the 30 h group was observed to have the best apparent metabolic rate for ether extract (p<0.05) and crude protein (p<0.05) and the highest activities of amylase, lipase and trypsin in small intestine. And the 24 h group was second only to the 30 h group in terms of the above two measures. Conclusion: These results indicated that the appropriate first feeding time of LingNan Yellow broilers was 24 to 30 hours after hatching.

• Journal of Dairy Science and Biotechnology
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• v.38 no.3
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• pp.161-167
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• 2020

Cheese pizzas fortified with casein phosphopeptide (CPP) and calcium were subjected to an in vitro digestion to assess whether CPP could prevent the precipitation of calcium. The total calcium content of the cheese pizzas was adjusted to 1,000 mg per pizza (~370 g) with the addition of calcium originating from eggshells. Two levels of trypsin-digested caseins (367 and 459 mg), with a CPP content of ~20%, were added to each pizza. The in vitro digested pizzas were then centrifuged and the supernatant was mixed with Na₂HPO₃ at 37℃ to estimate the possible soluble effect of CPP on calcium. After 24 h of reaction, the solution was centrifuged and the calcium content in the resultant supernatant was analyzed by inductively coupled plasma-atomic emission spectroscopy. One-way statistical analyses showed that CPP had a positive effect on the solubilization of calcium against phosphate (p<0.05). Cheese pizza supplemented with 459 mg of CPP powder was able to prevent precipitation of calcium by 98.8%, whereas no CPP-added cheese pizza solubilized 86.4% of the calcium. A sensory test was also carried out, revealing that panelists could not discern the bitter taste of the CPP added to the pizzas.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.6
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• pp.3975-3978
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• 2013

Background: Proteases play a regulatory role in a variety of pathologies including cancer, pancreatitis, thromboembolic disorders, viral infections and many others. One of the possible strategies to combat these pathologies seems to be the use of protease inhibitors. LC-pi I, II, III and IV (Lavatera cashmerian-protease inhibitors) have been found in vitro to strongly inhibit trypsin, chymotrypsin and elastase, proteases contributing to tumour invasion and metastasis, indicated possible anticancer effects. The purpose of this study was to check in vitro anticancer activity of these four inhibitors on human lung cancer cell lines. Material and Methods: In order to assess whether these inhibitors induced in vitro cytoxicity, SRB assay was conducted with THP-1 (leukemia), NCIH322 (lung) and Colo205, HCT-116 (colon) lines. Results: LC-pi I significantly inhibited the cell proliferation of all cells tested and also LC-pi II was active in all except HCT-116. Inhibition of cell growth by LC-pi III and IV was negligible. $IC_{50}$ values of LC-pi I and II for NCIH322, were less compared to other cell lines suggesting that lung cancer cells are more inhibited. Conclusion: These investigations might point to future preventive as well as curative solutions using plant protease inhibitors for various cancers, especially in the lung, hence warranting their further investigation.

• The Korean Journal of Community Living Science
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• v.27 no.4
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• pp.863-873
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• 2016

The current study was undertaken to determine the effects of the ethanol extracts of loquat (Eriobotrya japonica Lindl.) seeds, flesh or leaves on the differentiation of 3T3-L1 cells and male pig preadipocytes. The cell number was measured with the MTT assay after trypsin digestion. The cell differentiation was determined by measuring the glycerol-3-phosphate dehydrogenase (GPDH) activity and triglyceride(TG) content. No cytotoxicity was observed from the loquat flesh and leaf ethanol extracts at concentrations of 5, 10, 25, 50, 100 or $200{\mu}g/mL$ in 3T3-L1 cells and pig preadipocytes. However, the cell viability of neither cell line were affected by up $50{\mu}g/mL$ of loquat seed ethanol extract. Treatment with the loquat seed and leaf ethanol extracts significantly suppressed the terminal differentiation of both cell lines in a dose-dependent manner, as confirmed by the decrease in the glycerol-3-phosphate dehydrogenase(GPDH) activity and TG content. Treatment with the loquat seed and leaf ethanol extracts inhibited the GPDH activity and reduced the TG content of both cell types more effectively than that with the loquat flesh ethanol extract. The most potent anti-adipogenic effect was obtained in the case of the ethanol extract of loquat seeds.

• Food Science of Animal Resources
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• v.35 no.4
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• pp.473-478
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• 2015

The aim of this study was to screen and In vitro characterize the properties of bacteriocin produced by lactic acid bacteria isolated from Vietnamese fermented pork (Nem chua). One hundred and fifty LAB were isolated from ten samples of Nem chua and screened for bacteriocin-producing lactic acid bacteria. Antimicrobial activity of bacteriocin was carried out by spot on lawn method against both gram positive and gram negative bacteria. One isolate, assigned as KL-1, produced bacteriocin and showed inhibitory activity against Lactobacillus sakei, Leuconostoc mesenteroides and Enterococcus faecalis. To characterize the bacteriocin-producing strain, optimum temperature, incubation period for maximum bacteriocin production and identification of bacteriocin-producing strain were determined. It was found that the optimum cultivation temperature of the strain to produce the maximum bacteriocin activity (12,800 AU/mL) was obtained at 30℃. Meanwhile, bacteriocin production at 6,400 AU/mL was found when culturing the strain at 37℃ and 42℃. The isolate KL-1 was identified as L. plantarum. Antimicrobial activity of cell-free supernatant was completely inhibited by proteolytic enzyme of trypsin, alpha-chymotrypsin and proteinase K. Bacteriocin activity was stable at high temperature up to 100℃ for 10 min and at 4℃ storage for 2 d. However, the longer heating at 100℃ and 4℃ storage, its activity was reduced.

• YAKHAK HOEJI
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• v.51 no.4
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• pp.253-258
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• 2007

Bombycis corpus, a batryticated silkworm and white-stiff silkworm, is an oriental drug consisting of the dried larva of silkworm, dead and stiffened due to the infection of Beauveria. An peptidyl antimicrobial molecule was purified from B. corpus by reverse phase-column chromatography and HPLC. Its molecular weight was determined to be 2295.45 by using matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry. Its antimicrobial activity was diminished by trypsin digestion. It exhibited a broad antimicrobial spectrum against not only Gram-negative, but also Gram- positive bacteria. Furthermore, it was found to have an antimicrobial activity against vancomycin-resistant enterococci (VRE), methicillin-resistant S. arureus (MRSA), and vancomycin-intermediate resistant S. arureus (VISA). It may be a useful molecule for a new antibiotic development, especially against antibiotic resistant microbe. This substance may play a role in the defense system of this animal against Beauveria bassiana. This is the first report of a peptidyl antimicrobial substance from B. corpus.