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Inhibitory Effect of Camp Antagonist and Pka Inhibitors, and Stimulatory Effect of Adenylate Cyclase Agonist on Cathepsin K Processing in Cultured Mouse Osteoclasts (cAMP 길항제와 PKA 억제제 및 Adenylate Cyclase 촉진제의 백서 파골세포에서 Cathepsin K 생성에 대한 효과)

  • Shim, Youn-Soo
    • Journal of dental hygiene science
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    • v.6 no.1
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    • pp.1-9
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    • 2006
  • Cathepsin K (cat K) is the major cysteine protease expressed in osteoclasts and was thought to play a key role in matrix degradation during bone resorption. It was shown that the intracellular maturation of cat K was prevented by the cAMP antagonist, Rp-cAMP, and the protein kinase A (PKA) inhibitors of KT5720 and H89. In contrast, forskolin, a adenylate cyclase agonist, rather induced Cat K processing and maturation in osteoclasts. Furthermore, to determine whether cat K processing and maturation signaling involves protein kinase C (PKC), mouse total bone cells were treated with calphostin C, a specific inhibitor of PKC, however, no effect was observed, indicating that calphostin C did not affect to osteoclast-mediated cat K processing and maturation. Thus, it is indicated that the cAMP-PKA signaling pathway regulates cat K maturation in osteoclasts. Since secreted proenzymes have the potential to reenter the cell via M6P receptor, to prevent this possibility, it was tested cAMP antagonist Rp-cAMP and the PKA inhibitors KT5720 and H89 in the absence or presence of M6P. Inhibition of cat K processing by Rp-cAMP, KT5720, or H89 was observed in a dose-dependent manner. Furthermore, the addition of M6P resulted in enhanced potency of Rp-cAMP, KT5720 and H89. These dose-dependently inhibited in vitro bone resorption with a potency similar to that observed for inhibition of cat K processing.

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Quality and Antioxidative Characteristics of Cudrania tricuspidata Leaves Tea (꾸지뽕잎차의 품질 및 항산화 특성에 관한 연구)

  • Park, Bum-Ho;Back, Kyung-Yern;Lee, Sang-Il;Kim, Soon-Dong
    • Food Science and Preservation
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    • v.15 no.3
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    • pp.461-468
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    • 2008
  • To obtain basic da1a on the preparation of Cudrania tricuspidata leaves tea, the quality and anti-oxidative characteristics of dried raw leaves (RT), pan-fired leaves tea (PT) and fermented leaves tea (FT) were investigated. General characteristics of RT, PT and FT, respectively, were: moisture content 18.47, 6.23 and 8.50%; crude protein content 17.77, 20.46 and 19.13%; and carbohydrate content 54.42, 62.52 and 61.96%. The crude lipid and ash contents were in the range 0.05 - 0.07% and 9.27 -10.74% respectively; the water soluble solid content was in the order FT > PT > RT and ranged from 23.10 - 37.38%; there were no significant differences in the total polyphenol content (815.24 - 835.16 mg%). Although $L^*$ values of PT (20.94) and FT (20.85) were lower than those of RT (34.71), the $a^*$ value in PT and the $b^*$ value in FT were highest. In all ethanol extracts the reducing power, electron-donating ability and superoxide dismutase (SOD)-like activity increased in a concentration-dependent manner. Furthermore, the activity in FT was higher than in PT or RT. The total free amino-acid content was higher in FT (1429.93 mg%) than RT (1108.94 mg%) or PT (833.13 mg%). The major amino acids were L-asparagine and L-valine in RT, L-cysteine and L-glutamic acid in PT and L-proline in FT. In a sensory evaluation of PT and FT, bitter and astringent tastes were decreased relative to RT, while sweet and savory tastes, flavor, color and overall acceptability were increased. These results indicate that FT bas a higher antioxidant effect and free-amino-acid content than PT, while the sensory quality of FT is similar to that of PT.

Chungkook-jang Koji Fermentation with Rice Straw (볏짚을 이용(利用)한 청국장 제조(製造)에 관(關)한 연구(硏究))

  • Kim, Kyung-Ja;Ryu, Myung-Ki;Kim, Sang-Soon
    • Korean Journal of Food Science and Technology
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    • v.14 no.4
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    • pp.301-308
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    • 1982
  • Chungkook-jang Koji was fermented with rice straw at $40^{\circ}C\;and\;50^{\circ}C$ for 72 hours. The changes of proximate composition, pH, titrable acidity, nitrogen compounds, protease activity and free-amino acids during the fermentation were investigated. Moisture, lipid and protein contents remained essentially unchanged during the fermentation. The pH was gradually increased from 6.4 to 7.46 and 7.82 at $40^{\circ}C\;and\;50^{\circ}C$, respectively, after 72 hour fermentation. Amino type and water soluble nitrogen increased as fermentation progressed. however, the former slightly decreased after 60 hour fermentation. Chungkook-jang fermented at $40^{\circ}C$ showed somewhat higher protease activity than $50^{\circ}C$. However, protease activity at both fermentation temperatures showed the same trend; that is, it increased until 48 hour fermentation and thereafter decrease. Free amino acid content of Chung-kook-jang after 72 hour fermentation at $40^{\circ}C$ was 6 times greater than that of the steamed soybean, while it was 2.5 times greater at $50^{\circ}C$. Based on these results. it seems that the optimum fermentation conditions for Chungkook-jang were $40^{\circ}C$ and 72 hours.

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Antioxidant and anti-inflammatory effects of red garlic compositions (홍마늘 조성물의 항산화 및 항염증 효과)

  • Kang, Min Jung;Kim, Dong-Gyu;Shin, Jung Hye
    • Food Science and Preservation
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    • v.24 no.3
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    • pp.446-454
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    • 2017
  • Garlic (Allium sativum L.) and traditional herb has several functional properties and strong biological activities, making it useful as a functional food material. We investigated the antioxidant and anti-inflammatory activity of mixed compounds from red garlic and supplementary materials, including ginger (Zingiber officinale Roscoe), doraji (Platycodon grandiflorum), quince (Chaenomeles sinensis), citrus peel (Citri Pericarpium), and mint (Mentha arvensis). The extracts were prepared with water (W) and ethanol (E) at $70^{\circ}C$ (W-70, E-70) and $95^{\circ}C$ (W-95, E-95) for 3 h. The total content of phenolic compounds was the highest in E-70 (608.60 mg/100 g). Alliin, one of the active ingredients in red garlic, was contained at 1.18-1.29 mg/g and 0.81-0.85 mg/g in water and ethanol extract, respectively. Another active ingredient of red garlic, S-allyl-cysteine (SAC) had higher content in the water extract than in the ethanol extracts. DPPH radical scavenging activity was higher in E-70 (15.96-73.65%) at $313-5,000{\mu}g/mL$. ABTS radical scavenging activity was also higher in E-70 (5.71-77.19%) than in the others. The ROS production rate showed the same tendency as the NO production, with more efficacy in E-95. The expression level of iNOS and $IL-1{\beta}$ was decreased in the E-95 significantly at the concentration of $1,000{\mu}g/mL$ compared to the lipopolysaccharide (LPS) treated group. Based on the above results, the antioxidative and anti-inflammatory activities of the extracts of red garlic and supplementary materials were expressed by different useful substances. The contents of these useful substances were different according to the extraction solvent and temperature.

Relationship between Reactive Oxygen Species and Adenosine Monophosphate-activated Protein Kinase Signaling in Apoptosis Induction of Human Breast Adenocarcinoma MDA-MB-231 Cells by Ethanol Extract of Citrus unshiu Peel (진피 추출물에 의한 인간유방암 MDA-MB-231 세포의 apoptosis 유도에서 ROS 및 AMPK의 역할)

  • Kim, Min Yeong;HwangBo, Hyun;Ji, Seon Yeong;Hong, Su-Hyun;Choi, Sung Hyun;Kim, Sung Ok;Park, Cheol;Choi, Yung Hyun
    • Journal of Life Science
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    • v.29 no.4
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    • pp.410-420
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    • 2019
  • Citrus unshiu peel extracts possess a variety of beneficial effects, and studies on their anticancer activity have been reported. However, the exact mechanisms underlying this activity remain unclear. In the current study, the apoptotic effect of ethanol extract of C. unshiu peel (EECU) on human breast adenocarcinoma MDA-MB-231 cells and related mechanisms were investigated. The results showed that the survival rate of MDA-MB-231 cells treated with EECU was significantly inhibited in a concentration-dependent manner, which was associated with the induction of apoptosis. EECU-induced apoptosis was associated with the activation of caspase-8 and caspase-9, which initiate extrinsic and intrinsic apoptosis pathways, respectively, and caspase-3, a representative effect caspase. EECU suppressed the expression of the inhibitor of apoptosis family of proteins, leading to an increased Bax/Bcl-2 ratio and proteolytic degradation of poly (ADP-ribose) polymerase. EECU also enhanced the loss of the mitochondrial membrane potential and cytochrome c release from the mitochondria to the cytosol, along with truncation of Bid. In addition, EECU activated AMP-activated protein kinase (AMPK), and compound C, an AMPK inhibitor, significantly weakened EECU-induced apoptosis and cell viability reduction. Furthermore, EECU promoted the generation of reactive oxygen species (ROS), which acted as upstream signals for AMPK activation as pretreatment of cells, with the antioxidant N-acetyl cysteine reversing both EECU-induced AMPK activation and apoptosis. Collectively, these findings suggest that EECU inhibits MDA-MB-231 adenocarcinoma cell proliferation by activating intrinsic and extrinsic apoptotic pathways, which was mediated through ROS/AMPK-dependent pathways.

Effects of Fresh and Black Garlic Hot Water Extract Powder on the Lipid Composition of Hypercholesterolemia Rats (생마늘 및 흑마늘 열수추출 분말이 고콜레스테롤 혈증 흰쥐의 체내 지질조성에 미치는 영향)

  • Kang, Min-Jung;Shin, Jeong Yeon;Lee, Soo Jung;Shin, Jung Hye
    • Journal of Life Science
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    • v.31 no.1
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    • pp.37-46
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    • 2021
  • The effects of freeze-dried powder from fresh and black garlic hot water extracts on the lipid metabolism in Sprague-Dawley rats fed a high-cholesterol diet were analyzed. The experimental group was classified into the normal group (NG), the high-fat (HF) and high-cholesterol diet group (CG), the HFC and 1% fresh garlic hot water extract powder-added diet group (FGEG), and the HFC and 1% black garlic hot water extract powder-added diet group (BGEG), respectively. The serum total lipid content was 381.52±7.30 mg/ml and 368.80±4.40 mg/ml in the FGEG and the BGEG, respectively, and was significantly lower than that of the CG. The total cholesterol and triglyceride contents of the FGEG and BGEG were also significantly lower than that of the CG. The high-density lipoprotein (HDL) cholesterol was significantly higher, and the low-density lipoprotein (LDL) and the very low-density lipoprotein (VLDL) cholesterol content was lower in the FGEG and BGEG than in the CG. The serum ALT and AST activities were significantly lower than those of the CG, and especially the BGEG was lower. The total cholesterol content and the triglyceride levels of the liver tissue were 36.0% and 14.3% lower in the BGEG than in the CG, respectively. The thiobarbituric acid reactive substance (TBARS) concentrations in the serum and the liver tissue were higher in the CG than in the FGEG and BGEG, but there was no difference between them. Based on these results, garlic extract powders significantly reduced the lipid profile and increased the antioxidant activity in rats in vivo. The black garlic hot water extract powder was more effective than raw garlic because of the total number of phenolic compounds and browning substances in the black garlic.

The Induction of ROS-dependent Autophagy by Particulate Matter 2.5 and Hydrogen Peroxide in Human Lung Epithelial A549 Cells (미세먼지와 산화적 스트레스에 의한 인간 폐 상피 A549 세포에의 ROS 의존적 자가포식 유도)

  • Park, Beom Su;Kim, Da Hye;Hwangbo, Hyun;Lee, Hyesook;Hong, Su Hyun;Cheong, Jaehun;Choi, Yung Hyun
    • Journal of Life Science
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    • v.32 no.4
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    • pp.310-317
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    • 2022
  • Recently, interest in the harmful factors of particulate matter (PM), a major component of air pollution, has been increasing. In particular, PM2.5 with a diameter of less than 2.5 ㎛ is well known to induce oxidative stress accompanied by autophagy in human lung epithelial cells. However, studies on whether PM2.5 increases autophagy under oxidative stress and whether this process is reactive oxygen species (ROS)-dependent are insufficient. Therefore, in this study, we investigated whether PM2.5 promotes autophagy through the generation of ROS in human alveolar epithelial A594 cells. According to our results, cells co-treated with PM2.5 and hydrogen peroxide (H2O2) showed a lower cell viability than cells treated with each alone, which was associated with increased total and mitochondrial ROS production. The co-treatment of PM2.5 and H2O2 also increased autophagy induction, which was confirmed through Cyto-ID staining, and the expression of autophagy biomarker proteins increased. However, when ROS generation was artificially blocked by N-acetyl-L-cysteine pretreatment, the reduction in cell viability and induction of autophagy by PM2.5 and H2O2 co-treatment were markedly attenuated. Therefore, the present results suggest that PM2.5-induced ROS generation may play a critical role in autophagy induction in A549 cells.

The Ripening of Camembert Cheese Made with Mucor Miehei Rennet (Mucor Miehei 응유효소(凝乳酵素)로 제조(製造)한 Camembert Cheese의 숙성(熟成)에 관(關)한 연구(硏究))

  • Park, Mooh Il;Kim, Jong Woo
    • Korean Journal of Agricultural Science
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    • v.16 no.2
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    • pp.179-200
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    • 1989
  • Mucor miehei rennet(MR) was added as calf rennet(CR) substitutes in the fixed amounts of mixed rennets in making Camembert cheese. The conditions in the variations of chemical composition: water-soluble nitrogen, non-caseinic nitrogen, non-proteinic nitrogen, amino nitrogen, ammoniacal nitorgen, electrophoresis, molecular fractionation, mineral distribution, texture characterisitics, free amino acids and free fatty acids, were checked up with the sensory test and the chesse yields at each ripening period. The results obtained by investigating the utility of Mucor rennet were summarized as follows: 1. CR chesse, MR cheese and the mixed-rennet chesse failed to show any significant difference in their yields of 15%. 2. The contents of protein, fat and ash in MR cheese gave lower value than CR cheese did and with progress of ripening lactose decreased rapidly after 14 days of ripening. The difference among the rate of addition of mucor rennet was not recognized. 3. The WSN contents of 5 fresh sample chesse were from 14.7% to 17.3% and WSN increased from 39.7% to 41.0% with progress of ripening. After 21 days of ripening MR chesse had more WSN than CR cheese did. In NCN and ammoniacal nitrogen MR cheese showed higher value. 4. As the ripening progressed, MR chesse showed more cystein, phenylalanine and proline than CR chesse did but it failed to show any increase in aspartic acid, threonine and glutamic acid etc. 5. In the content of free fatty acid MR chesse showed higher value than CR cheese did and with the progress of ripening fatty acids increased from 8.36 mEq to 26.36 mEq but did not show any significant difference in the cheese types by the coagulant ratio. 6. Ca contents in the sample chesse were 0.238-0.27%, Mg 0.019-0.022%, Na 0.910-1.047%, and K 0.175-0.200%. The important non-sedimentable Ca in casein remained from 61 % to 77% without regard the ripening periods and added-rennets and Mg remained from 59.1% to 92.5% in non-sedimentable and water-soluble conditions. 7. In the fractionation of protein by ultrafilteration, MW> $5{\times}10^4$ decresed from 95% at the beginning period of ripening to 45% and MW< $10^4$ increased from 0.2% to 38% and definite caseinolysis was shown in all samples. 8. All the cheese showed to different electrophoretic patterns for the added-amounts of mucor rennet in the 14 days of ripenig. In the 28 days or ripening, MR cheese kept some bands on the patterns compared with CR cheese. 9. In vitro digestibility increased from 81.48-94.81 % to 94.47-98.61% but failed to show any significant difference in the cheese types by the coagulant ratio. 10. In hardness, MR cheese showed lower value compared with CR cheese as the ripening progressed. 11. The results of the sensory test failed to show any difference in flora rind, feelings in mouth and hands, deep structure, flavor and bitterness between CR Camembert cheese and MR Camembert chesse.

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Meat Quality and Nutritional Properties of Hanwoo and Imported Australian Beef (한우고기와 호주산 냉장수입육의 육질 및 영양성분 비교)

  • Cho, Soo-Hyun;Seong, Pil-Nam;Kang, Geun-Ho;Park, Beom-Young;Jung, Seok-Geun;Kang, Sun-Moon;Kim, Young-Chun;Kim, Jong-In;Kim, Dong-Hun
    • Food Science of Animal Resources
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    • v.31 no.5
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    • pp.772-781
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    • 2011
  • This study was conducted to investigate the proximate composition, meat color, Warner-Bratzler shear force (WBS), total collagen content (%), cooking loss (CL), fatty acid composition, amino acid composition, and mineral content of loin, strip loin, top round, and chuck tender Hanwoo beef and imported Australian beef. Protein content was significantly lower for strip loin and loin of Hanwoo QG $1^+$ and 1 beef when compared to that of Australian black Angus or cross beef (p < 0.05), whereas it was not significantly different for top round and chuck tender. Intramuscular fat content was higher for QG $1^+$ Hanwoo beef strip loin (15.48%) than that in Australian cross beef (8.83%) and it was also higher in loin for QG $1^+$ (17%) and 1 (15.52%) Hanwoo beef than that in Australian Angus beef (10.59%) and cross beef (9.21%) (p < 0.05). The CIE $L^*$ value was significantly higher for Australian cross beef strip loin than that of Hanwoo beef but the CIE $a^*$ value was not significantly different between the same cuts from different origins. However, CIE $b^*$ values were significantly higher for the Australian cross beef than those for four cuts of Hanwoo beef and Australian Angus beef (p < 0.05). No significant difference in WBS of loin, top round, or chuck tender was observed among the different beef sample origins except that strip loin had significant higher WBS for Australian cross beef (3.02 kg) but lower for Australian Angus beef (2.13 kg). Australian cross beef contained significantly higher palmitic acid, stearic acid, linolenic acid, but lower palmitoleic acid, oleic acid, eicosenoic acid content in strip loin, loin, and top round than those of QG $1^+$ and 1 Hanwoo beef (p < 0.05). QG $1^+$ Hanwoo beef had significant lower saturated fatty acid and higher monounsaturated fatty acid content than those in Australian cross beef (p < 0.05). Hanwoo $1^+$ beef had more glycine in top round and chuck tender, whereas Hanwoo QG 1 beef and Australian Angus beef had higher cysteine, methione, and glycine levels in strip loin and loin and valine and leucine in top round and chuck tender than the same cuts of Hanwoo $1^+$ beef (p < 0.05). No significant differences were observed for Ca, Fe, or Zn content among the different beef samples.