• Title/Summary/Keyword: Trp-P-1

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Antimutagenic and Anticancer Effects of Ethanol Extract from Korean Traditional Doenjang Added Sea Tangle (다시마 분말을 첨가한 전통된장 에탄을 추출물의 항돌연변이성 및 항암효과)

  • 최승필;이의용;이득식;함승시
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.2
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    • pp.322-328
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    • 2002
  • This study was carried out to investigate antimutagenic and anticancer effects of ethanolic extract of Korean traditional doenjang added sea tangle. Most of the mineral content of doeniang was increased by addition of sea tangle. In the Ames test, the antimutagenic effect of ethanol extract of Korean traditional doenjang added 5% sea tangle was higher than that of control (no additive), 10%, and 15% sea tangle additions. The inhibition rate of ethanol extract (200$\mu\textrm{g}$/plate) of doenjang added 5% sea tangle in the S. typhimurium TA100 strain showed 97.0% inhibition against the mutagenesis induced by MNNG. In addition, the suppression of ethanol extract (200$\mu\textrm{g}$/plate) of doenjang added 5% sea tangle in the S. typhimurium TA98 and TA100 strains showed 60.2% and 69.1% inhibition respectively, against the mutagenesis induced by 4NQO. The suppressions under the same condition against B($\alpha$ )P and Trp-P-1 in the TA98 and TA100 strains were 71.7% and 87.3%, and 66.6% and 80.8%, respectively. In the anticancer effects, the cytotoxicity of doenjang added 5% sea tangle on the cell lines with human lung carcinoma (A549), human hepatocellular carcinoma (HepG2), and human gastric carcinoma (KATOIII) were inhibited with increasing the extract concentration. The treatment of 1.0 mg/mL Doenjang added 5% sea tangle showed strong cytotoxicity of 56.4%, 87.67%, and 89.5% against A549, HepG2, and KATOIII, respectively.

Effects of Casein and Protein-free Diets on Endogenous Amino Acid Losses in Pigs

  • Zhang, Yongcheng;Li, Defa;Fan, Shijun;Piao, Xiangshu;Wang, Jitan;Han, In K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.11
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    • pp.1634-1638
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    • 2002
  • Quantification of endogenous amino acid loss at the terminal ileum is an essential means for calculation of the true amino acid digestibility of a feedstuff. Since nitrogen appeared in the determined diet or not could shift the results very much, also, none of digestibility markers could be recovered with 100% rate at the terminal ileum, the objectives of the present study were: (1) to determine endogenous amino acid losses when fed either a casein diet or a protein-free diet and (2) to examine the reliability of chromic oxide or acid insoluble ash in the protein-free diet. Six ileal-cannulated pigs ($65{\pm}1.85 kg$ BW) with a simple T-cannula in the terminal ileum were used in a replicated $3{\times}3$ Latin square designed trial, after allowed a 14 d recuperation period. Each test period ran for 12 days comprised of a 10 d adjustment period and a 2 d collection period. The endogenous AA losses of His, Ile, Lys, Cys, Thr, Val, Trp, Asp, Glu, and Ser from pigs fed the casein diet were significantly higher than those of the protein-free diet (p<0.05). No significant difference was found in the amount of endogenous amino acid loss when determined with the different markers in the protein-free diet (p>0.05). These data suggest that endogenous amino acid loss could be underestimated when a protein-free diet is used. A direct effect of dietary peptides on the endogenous amino acid loss was found when the casein diet was fed. Our results also indicate that acid insoluble ash can be used as an inert marker as an alternative to chromic oxide when measuring endogenous amino acid loss.

Site-specific and deletional mutagenesis for two regions of Verotoxin-2 A gene encoding enzymatically active domain (Verotoxin-2 A 유전자의 효소활성 부위에 대한 위치특이적 변이 및 결손변이유발)

  • Kim, Yong-hwan;Kim, Sang-hyun;Cha, In-ho;Kim, Kyoung-shook;Lee, Young-choon
    • Korean Journal of Veterinary Research
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    • v.37 no.3
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    • pp.541-546
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    • 1997
  • There are two conserved regions with a significantly high amino acid sequence homology among the A subunits of STX, SLTs and ricin. To produce an inactive Verotoxin-2 (VT-2), two different mutants, pE167D and pDE5A, were constructed by site-directed mutagenesis, respectively, on the basis of the previous reports that two regions lie within the active-site clefts of the A subunits of ricin and STX family. The cytotoxicity ($10^3$ $CD_{50}/ml$) of VT-2 holotoxin with E167D mutation was reduced by $10^3$-fold compared with wild-type level. In addition, VT-2 with DE5A ($Trp_{202}GlyArgIleSer_{206}$) deletion mutation showed a significantly low cytotoxicity ($10^1$ $CD_{50}/ml$), resulting in $10^5$- and $10^2$-fold reductions, respectively, compared with the wild-type and E167D mutatant. SDS-PAGE for protein samples showed a 33-kDa band corresponding to the A subunit of VT-2. These results indicate that reduction in cytotoxic activity was affected not by amount of VT-2 protein produced but by mutation.

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Antimutagenic and Cytotoxic Effects of Acer ginnala Max. Bark Extracts (신나무 껍질 추출물의 항돌연변이원성 및 세포독성 효과)

  • Oh Heung-Seok;Cui Cheng-Bi;Choi Hyung-Taek;Kim Soo-Hyun;Jeon Mi-Sun;Ham Seung-Shi
    • Food Science and Preservation
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    • v.11 no.4
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    • pp.550-556
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    • 2004
  • In the present study, we investigated the antimutagenic and cytotoxic effects of Acer ginnala Max. bark extract on S. typhimurium TA98, TA100 and cancer cell lines with Ames test and SRB assay, respectively. They were extracted with methanol and then fractionated using hexane, chloroform, ethyl acetate, butanol, and water to obtain the fractions. The inhibition rate of methanol ($200\;{\mu}g/plate$) of Acer ginnala Max. bark extract in the Salmonella typhimurium TA100 strain showed $83.3\%$ against the mutagenesis induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). In addition, the suppression of methanol extract with same concentration of in the Salmonella typhimurium TA98 and TA100 strains showed $80.3\%\;and\;92.7\%$ inhibition against 3-amino-1,4-dimethyl-5H-pyrido-(4,3-b)indol (Trp-P-1), respectively. The cytotoxicity effects of Acer ginnala Max. bark extract against the cell lines with human lung carcinoma (A549), human gastric carcinoma (AGS), human hepatocellular carcinoma (Hep3B) and human breast adenocarcinoma (MCF-7) were inhibited with the increase of the extract concentration. The treatment of 1.0 mg/mL Acer ginnala Max. bark methanol extract of methanol showed strong cytotoxicities of $77.3\%,\;90.4\%,\;88.9\%,\;and\;83.7\%$ against A549, AGS, Hep3B and MCF-7, respectively.

Assessment of Biomarkers in Acetaminophen-Induced Hepatic Toxicity by siRNA

  • Kang, Jin-Seok;Yum, Young-Na;Kim, Joo-Hwan;Park, Sue-Nie
    • Biomolecules & Therapeutics
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    • v.17 no.4
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    • pp.438-445
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    • 2009
  • We investigated global gene expression from both mouse liver and mouse hepatic cell lines treated with acetaminophen (APAP) in order to compare in vivo and in vitro profiles and to assess the feasibility of the two systems. During our analyses of gene expression profiles, we picked up several down-regulated genes, such as the cytochrome P450 family 51 (Cyp51), sulfotransferase family cytosolic 1C member 2 (Sult1c2), 3-hydroxy-3-methylglutaryl-Coenzyme A synthase 1 (Hmgcs1), and several genes that were up-regulated by APAP, such as growth arrest and DNA-damage-inducible 45 alpha (Gadd45a), transformation related protein 53 inducible nuclear protein 1 (Trp53inp1) and zinc finger protein 688 (Zfp688). For validation of gene function, synthesized short interfering RNAs (siRNAs) for these genes were transfected in a mouse hepatic cell line, BNL CL.2, for investigation of cell viability and mRNA expression level. We found that siRNA transfection of these genes induced down-regulation of respective mRNA expression and decreased cell viability. siRNA transfection for Cyp51 and others induced morphological alterations, such as membrane thickening and nuclear condensation. Taken together, siRNA transfection of these six genes decreased cell viability and induced alteration in cellular morphology, along with effective inhibition of respective mRNA, suggesting that these genes could be associated with APAP-induced toxicity. Furthermore, these genes may be used in the investigation of hepatotoxicity, for better understanding of its mechanism.

Naringenin stimulates cholecystokinin secretion in STC-1 cells

  • Park, Min;Kim, Kyong;Lee, Yu Mi;Rhyu, Mee Ra;Kim, Hye Young
    • Nutrition Research and Practice
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    • v.8 no.2
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    • pp.146-150
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    • 2014
  • BACKGROUND/OBJECTIVES: Cholecystokinin (CCK), a hormone or neuropeptide, is secreted in response to intraluminal nutrients by enteroendocrine I-cells of the intestine and has important physiological actions related to appetite regulation and satiety. The stimulation on CCK secretion from the intestine is of potential relevance for body weight management. Naringenin (4',5,7-trihydroxyflavanone) and its glycoside naringin (naringenin 7-rhamnoglucoside) have been reported to have many biological functions. In the current study, we investigated the question of whether naringenin and naringin could stimulate CCK secretion and then examined the mechanisms involved in CCK release. MATERIALS/METHODS: STC-1 cells were used as a model of enteroendocrine cells. CCK release and changes in intracellular $Ca^{2+}$ ($[Ca^{2+}]_i$) were measured after incubation of cells with naringenin and naringin for 1 h. RESULTS: Naringenin caused significant (P < 0.05) stimulation of CCK secretion, but naringin did not. In addition, regarding the secretory mechanisms, naringenin-induced CCK secretion involved increases in $[Ca^{2+}]_i$, influx of extracellular $Ca^{2+}$, at least in part, and activation of TRP channels, including TRPA1. CONCLUSION: Findings of this study suggest that naringenin could have a role in appetite regulation and satiety.

Polymorphisms in XRCC1 Gene, Alcohol drinking, and Risk of Colorectal Cancer: a Case-control Study in Jiangsu Province of China

  • Gao, Chang-Ming;Ding, Jian-Hua;Li, Su-Ping;Liu, Yan-Ting;Cao, Hai-Xia;Wu, Jian-Zhong;Tang, Jin-Hai;Tajima, Kazuo
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.11
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    • pp.6613-6618
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    • 2013
  • To evaluate the relationship between alcohol drinking, XRCC1 codon 194 and 399 polymorphisms and risk of colorectal cancer, we conducted a case-control study with 315 colorectal cancer cases (105 colon, 210 rectal) and 439 population-based controls in Jiangsu Province of China. The XRCC1 codon 194 and 399 genotypes were identified using polymerase chain reaction and restrictrion fragment length polymorphism methods (PCR-RFLP). A structured questionnaire was used to elicit detailed information. Odds ratios (ORs) were estimated with an unconditional logistic model. In this study no significant differences were observed among the studied groups with regard to the genotype distribution of the XRCC1 codons 194 and 399 and the risk of colorectal cancer did not appear to be significantly influenced by genotype alone, whereas alcohol consumption showed a positive association (P for trend <0.01). When combined effects of XRCC1 polymorphisms and alcohol consumption were analyzed, we found that the 194Trp or 399Gln alleles further increased the colorectal cancer risk due to high alcohol intake. These findings support the conclusion that colorectal cancer susceptibility may be altered by gene-environment interactions.

Interaction between a Blood Vessel-Inducing Protein Angiogenin and Its Binding Protein Actin

  • Chang, Soo-Ik;Paik, Seung-Bum;So, Seung-Ho;Ahn, Byung-Cheol
    • BMB Reports
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    • v.29 no.4
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    • pp.353-358
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    • 1996
  • Bovine angiogenin (bAng) is a potent blood vessel inducing protein purified from cow In ilk. fluorescence spectroscopy has been used to study the interaction of bAng with actin in 50 mM Tris-HCl pH 7.5, and 1 mM $CaCl_2$ at $25^{\circ}C$. Actin contains four tryptophans but bAng contains no tryptophans. A 50% decrease in intrinsic fluorescence accompanied formation of the bAng/actin complex. By contrast, the interaction of RNase A, a homologous protein to bAng, with actin results in about 10% quenching of the fluorescence. Fluorescence titration experiments were performed by adding increasing concentrations of bAng (0~1.0 ${\mu}M$) to a constant concentration of actin (0.1 ${\mu}M$), and the dissociation constant $K_d$ for the bAng/actin complex and the stoichiometry n were measured as $20{\pm}1$ nM and $1.0{\pm}0.1$ respectively. These results suggest that the interaction between bAng with actin is specific and that quenching of actin fluorescence has occurred in the bAng/actin complex. The bAng binding sites of actin are discussed in the results of this study, and we propose that Trp-80 in the small domain of bovine actin is responsible for the bAng/actin binding.

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Effects of Low Crude Protein Diets Supplemented with Synthetic Amino Acids on Performance, Nutrient Utilization and Carcass Characteristics in Finishing Pigs Reared Using a Phase Feeding Regimen

  • Lee, J.H.;Kim, J.H.;Kim, J.D.;Kim, S.W.;Han, In K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.5
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    • pp.655-667
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    • 2001
  • This experiment was conducted to investigate the effect of feeding a low CP diet supplemented with synthetic amino acids on performance, nutrient utilization and carcass characteristics of finishing pigs fed under a three-phase feeding regimen. Ninety-six finishing pigs (Landrace$\times$Large White$\times$Duroc), $55.75kg{\pm}0.65$ of initial body weight, were blocked by weight and sex and allotted to four dietary treatments in a randomized block design. There were six pens per treatment and four pigs per pen. Pigs were fed a 16%-14%-12% CP (for phase I-II-III, respectively), sequence of diets. Dietary treatments were 1) Control, 2) Con+L (a sequence of diets reduced in CP by l percentage unit with lysine (L) supplementation, 3) Con+LMT (a sequence of diets reduced in CP by 2 percentage unit with LYS, methionine (MET) and threonine (THE) supplementation) and 4) Con+LMTT (a sequence of diets reduced in CP by 3 percentage unit with LYS, MET, THR and tryptophan (TRP) supplementation). The finishing period (55 to 105 kg) was divided into three phases (55 to 72 kg, 72 to 90 kg and 90 to 105 kg). Pigs fed either the control or Con+L diet grew faster (p<0.05) than pigs fed the Con-LMT or Con+LMTT diet. There was no difference in ADFI among dietary treatments. Phosphorus (P) digestibility was lowest in the control group and highest in the Con+LMTT group (p<0.05). Within each phase, no significant differences in dry matter (DM) and CP digestibilities were found. Although some amino acid digestibilities were affected by dietary treatments, digestibilities of essential amino acids (EAA), non-essential amino acids (NEAA) and total amino acid were not significantly influenced by dietary treatments. For the entire experiment periods, Con+L, Con+LMT and Con+LMTT treatments resulted in 13.4, 18.8 and 21.6% lower total N excretion compared with the control. Con+LMT and Con+LMTT treatments showed significantly lower BUN concentration compared with the control and Con+L treatment (p<0.05), but there was no significant difference in BUN concentration between pigs fed the control and Con+L treatment or between pigs fed Con+LMT and Con+LMTT treatments (p>0.05). Carcass length, backfat thickness and carcass grade were not significantly affected by dietary treatments (p>0.05). In conclusion, reducing dietary CP level by 1 percentage unit and supplementing only LYS at each phase could be a very beneficial feeding strategy for finishing pigs fed under a three phase feeding regimen in terms of both environmental and economical aspects.

Rumen Degradability and Post-ruminal Digestion of Dry Matter, Nitrogen and Amino Acids of Three Protein Supplements

  • Gao, Wei;Chen, Aodong;Zhang, Bowen;Kong, Ping;Liu, Chenli;Zhao, Jie
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.4
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    • pp.485-493
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    • 2015
  • This study evaluated the in situ ruminal degradability, and subsequent small intestinal digestibility (SID) of dry matter, crude protein (CP), and amino acids (AA) of cottonseed meal (CSM), sunflower seed meal (SFSM) and distillers dried grains with solubles (DDGS) by using the modified three-step in vitro procedure. The ruminal degradability and subsequent SID of AA in rumen-undegradable protein (RUP-AA) varied among three protein supplements. The result show that the effective degradability of DM for SFSM, CSM, and DDGS was 60.8%, 56.4%, and 41.0% and their ruminal fermentable organic matter was 60.0%, 55.9%, and 39.9%, respectively. The ruminal degradable protein (RDP) content in CP for SFSM, CSM, and DDGS was 68.3%, 39.0%, and 32.9%, respectively, at the ruminal solid passage rate of 1.84%/h. The SFSM is a good source of RDP for rumen micro-organisms; however, the SID of RUP of SFSM was lower. The DDGS and CSM are good sources of RUP for lambs to digest in the small intestine to complement ruminal microbial AA of growing lambs. Individual RUP-AA from each protein source was selectively removed by the rumen microorganisms, especially for Trp, Arg, His, and Lys (p<0.01). The SID of individual RUP-AA was different within specific RUP origin (p<0.01). Limiting amino acid was Leu for RUP of CSM and Lys for both RUP of SFSM and DDGS, respectively. Therefore, different protein supplements with specific limitations should be selected and combined carefully in growing lambs ration to optimize AA balance.