• Title/Summary/Keyword: CAT protein

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Effects of Edible Lentinus tuber-regium on Oxidative Stress and Defense System in Serum of SD Rats (유용버섯 Lentinus tuber-regium이 산화적 스트레스 및 방어체계에 미치는 영향)

  • Choi, Jin-Ho;Park, Soo-Hyun;Kim, Dae-Ik;Kim, Jeung-Min;Kim, Chang-Mok;Kim, Gwang-Po
    • The Korean Journal of Mycology
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    • v.29 no.1
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    • pp.41-46
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    • 2001
  • Oxidative stress and defense system of SD-rats were studied with an edible Nigerian mushroom, namely, Lentinus tuber-regium (Fries) Singer. Experimental diets prepared with Lentinus tuber-regium (LTR) instead of carbohydrates were fed to SD rats for 6 weeks. Hydrolxyl radical $({\cdot}OH)$ formations were significantly inhibited (21.7% and 16.4%, respectively). In LTR-50 and LTR-100 groups used instead of carbohydrates, and hydrogen peroxide and nitric oxide (NO) were also significantly inhibited by 10%, and $6{\sim}10%$, respectively compared with control group, but there was no significant changes in superoxide radical $({O_2}^-)$ formations in these groups. Lipid peroxide (LPO) and oxidized protein (OP) levels as an oxidative stress were desirably inhibited ($6{\sim}12%\;and\;5{\sim}13%$, respectively) in these LTR groups compared with control group. Superoxide dismutase (SOD), glutathione peroxidase (GSHPx) and catalase (CAT) activities were significantly increased ($15{\sim}50%,\;10{\sim}25%\;and\;60{\sim}90%$, respectively) in these LTR groups. These results suggest that an edible mushroom, Lentinus tuber-regium may inhibit an oxygen radicals and oxidative stresses, but may also effectively modulate an aging processes.

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Effects of low salinity stresses on the physiology of disc abalone, Haliotis discus discus (저염분 자극에 의한 둥근전복, Haliotis discus discus의 생리학적 변화)

  • Jwa, Min-Seok;Kang, Kyung-pil;Choi, Mi-Kyung;Yeo, In-Kyu
    • Journal of fish pathology
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    • v.22 no.3
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    • pp.293-303
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    • 2009
  • Effects of stress on the low salinity stress were examined in the pacific abalone Haliotis discus discus. Changes in survival rate, hemolymph count, antioxidant enzyme activities (catalase: CAT and superoxide dismutase: SOD), respiratory burst activity, phenoloxidase activity, lysozyme activity and expression of heat shock protein 70 (HSP70) mRNA were measured 0, 3, 6, 12, 24 or 48hours after low salinity treatment with 25, 30, 33 and 35 psu. Survival rates of pacific abalone were 100% at 33 and 35 psu, but 93 and 97% at 25 and 30 psu for 48 hours, respectively. Hemolymph counts decreased in the time elapsed-dependent way at all of the experimental groups. At low salinity, 25 and 30 psu, SOD and CAT activity increased compared to the experimental group of 33 psu. Moreover, respiratory burst activities of the pacific abalone seemed to have no effect on low salinity stress at any experimental group. However, phenoloxidase activity is an important component of the defence against pathogen that was decreased in a reduction of salinity dependent way. Lysozyme activity also immediately reduced at 25 psu experimental group for 48 h. The HSP70 mRNA was weakly expressed at 33 psu, but strongly detectable at 25 psu experimental group. The HSP 70 mRNA expression in gill increased in the time elapsed-dependent way at 25 psu experimental group and then recovered at 48 h. These results suggest that low salinity stress give rise to inhibitory action of immune system as a result of the decrease of phenoloxidase and lysozyme activity in the pacific abalone, especially.

Effects of Nutrition Education in Type 2 Diabetes Mellitus on Diabetes Control and Blood Antioxidant Status (제2형 당뇨환자에 대한 영양교육이 당뇨병 관리와 혈액 항산화 상태에 미치는 영향)

  • Shin, Kyung-Nam;Lee, Hye-Sang;Kwon, Chong-Suk
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.5
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    • pp.689-695
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    • 2011
  • Diabetic patients need nutritional education more than those suffering from other diseases because of the necessity of controlling blood glucose levels with dietary treatment. The purpose of this study was to find out the effectiveness of nutrition education on diabetes control and antioxidant status, both of which are related to diabetic complications. Thirty (15 males and 15 females) type 2 diabetes mellitus patients aged $66.7{\pm}8.8$ years participated in a 4-week nutrition education program. Nutrient intakes, blood glucose level, antioxidant status, and DNA damage were evaluated before, immediately after, and three months after the education program. Changes in those parameters over time were analyzed using repeated-measures analysis of covariance. Over time, HbA1c (p=0.000), plasma total cholesterol (p=0.002), plasma thiobarbituric acid related substances (TBARS; p=0.000), and leukocyte DNA damage (p=0.000) significantly decreased; plasma retinol (p=0.001), plasma tocopherol (p=0.000), erythrocyte catalase (CAT; p=0.000), and erythrocyte glutathione peroxidase (GPx; p=0.000) significantly increased. In an evaluation of nutrient intakes by Dietary Reference Intakes for Koreans (KDRI), energy (p=0.009), phosphorus (p=0.033), sodium (p=0.001), potassium (p=0.019), zinc (p=0.043), riboflavin (p=0.050), folic acid (p=0.048) and vitamin C (p=0.008) intakes had significant positive changes. In a correlation analysis of the biochemical and nutritional changes resulting from the education program, plasma TBARS were negatively correlated with potassium (r=-0.418, p<0.05), iron (r=-0.443, p<0.05), riboflavin (r=-0.432, p<0.05), and folic acid (r=-0.446, p<0.05) intakes, while plasma retinol was positively correlated with energy (r=0.543, p<0.01), protein (r=0.543, p<0.01), phosphorus (r=0.425, p<0.05), iron (r=0.485, p<0.05), zinc (r=0.570, p<0.01) and niacin (r=0.510, p<0.05) intakes. Erythrocyte CAT was positively correlated with folic acid intake (r=0.605, p<0.01). From these results, we suggest that an improvement in nutrition resulting from a diabetic education program for type 2 diabetes patients led to improvement in their antioxidant status, also possibly reducing complications resulting from diabetes.

Cooperation of $G{\beta}$ and $G_{\alpha}q$ Protein in Contractile Response of Cat Lower Esophageal Sphincter (LES)

  • Sohn, Uy-Dong;Lee, Tai-Sang
    • The Korean Journal of Physiology and Pharmacology
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    • v.7 no.6
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    • pp.349-355
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    • 2003
  • We previously shown that LES contraction depends on $M_3$ receptors linked to PTX insensitive $G_q$ protein and activation of PLC. This results in production of $IP_3$, which mediates calcium release, and contraction through a CaM dependent pathway. In the esophagus ACh activates $M_2$ receptors linked to PTX sensitive $G_{i3}$ protein, resulting in activation of PLD, presumably, production of DAG. We investigated the role of PLC isozymes which can be activated by $G_q$ or $G{\beta}$ protein on ACh-induced contraction in LES and esophagus. Immunoblot analysis showed the presence of 3 types of PLC isozymes, $PLC-{\beta}1$, $PLC-{\beta}3$, and $PLC-{\gamma}1$, but not $PLC-{\beta}2$, $PLC-{\beta}4$, $PLC-{\gamma}2$, $PLC-{\delta}1$, and $PLC-{\delta}2$ from both LES and esophageal muscle. ACh produced contraction in a dose dependent manner in LES and esophageal muscle cells obtained by enzymatic digestion with collagenase. $PLC-{\beta}1$ or $PLC-{\beta}3$ antibody incubation reduced contraction in response to ACh in LES but not in esophageal permeabilized cells, but $PLC-{\gamma}1$ antibody incubation did not have an inhibitory effect. The inhibition by $PLC-{\beta}1$ or $PLC-{\beta}3$ antibody on Ach-induced contraction was antibody concentration dependent. The combination with $PLC-{\beta}_1$ and $PLC-{\beta}_3$ antibody completely abolished the contraction, suggesting that $PLC-{\beta}1$ and $PLC-{\beta}3$ have a synergism to inhibit the contraction in LES. $PLC-{\beta}1$, -${\beta}3$ or -${\gamma}1$ antibody did not reduce the contraction of LES cells in response to DAG ($10^{-6}$ M), suggesting that this isozyme of PLC may not activate PKC. When $G_{q/11}$ antibody was incubated, the inhibitory effect of the incubation of PLC ${\beta}3$, but not of PLC ${\beta}_1$ was additive (Fig. 6). In contrast, when $G_{\beta}$ antibody was incubated, the inhibitory effect of the incubation of PLC ${\beta}_1$, but not of PLC ${\beta}_3$ was additive. This data suggest that $G_{q/11}$/11 or $G{\beta}$ may activate cooperatively different PLC isozyme, $PLC{\beta}_1$ or $PLC{\beta}_3$ respectively.

Analysis of antigenic specificities of Puragonimus westeymuni developmental stages using immunoblot technique (Immunoblot technique을 이용한 폐흡충의 발육단계별 항원 특이성 분석)

  • 주경환;홍성철
    • Parasites, Hosts and Diseases
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    • v.27 no.1
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    • pp.1-8
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    • 1989
  • Serodiagnosis of parasitic infections is widely used, since parasites or their eggs are not always detected by ordinary methods. The sensitive tests such as ELISA are highly dependent on the purity of antigens used. To solve this problem. many workers have tried to find species-specific components of antigens, The present study was performed to determine the antigenic profile of crude saline extracts of 3, 5, 8 and 12-week old p. westermani worms, which were collected from experimentally infected cats, based on SDS-PAGE and immunoblot technique. The results were as follows: 1. The SDS-PAGE showed at least 30 Protein bands ranging from 229 kDa to 10 kDa molecular weight. The protein components of p. wsstermani changed chronologically during its developmental period. The 229 kDa band was recognized only in 12-week old worms ($$SEP_{l2}$). 2. Analysis by ELISA showed a significant increase in antibody levels at 3 weeks in infected cats using crude saline extract antigens ($SEP_3,{\;}SEP_5,{\;}SEP_8,{\;}SEP_{l2}$). 3. By EITB using $SEP_3$ and $SEP_5$ infected cats recognisea major protein bands with molecular weight of 60, 35, 28, 25 or 21 kDa at 3~12 weeks of infection, and 3 additional antigens, 19, 13 and 10 kDa, were detected at 8~12 weeks of infections. 4. Using $SEP_8$ 5 antigens, 91, 85, 31, 25 and 21 kDa, were consistently detected by all infected sera tested. In addition, 3 antigens of lg. 13 and 10 kDa were detected at 8~12 weeks of infection. Using $SEP_12$, similar results were obtained with that by using $SEP_8$ and 1 additional antigen of 229 kDa, specifically reacting with the sera from 12 weeks of in(traction, was recognized.

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Molecular Cloning and mRNA Expression of the Hanwoo CAT/enhancer-binding Protein α(C/EBPα) Gene (한우 CCAAT/enhancer-binding protein α(C/EBPα) 유전자의 동정과 mRNA의 발현)

  • Jeoung, Y.H.;Lee, S.M.;Park, H.Y.;Yoon, D.H.;Moon, S.J.;Chung, E.R.;Kang, M.J.
    • Journal of Animal Science and Technology
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    • v.46 no.6
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    • pp.909-916
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    • 2004
  • CCAAT/enhancer binding proteins(C/EBP) are a group of transcription factors expressed during preadipocyte differentiation. In the C/EBPs, C/EBPa plays an important role in lipid deposition and adipocyte differentiation. In this studies, we report the identification, characterization, and expression of a Hanwoo CIEBP$\alpha$ The Hanwoo C/EBP$\alpha$DNA includes a 1059 bp open reading frame encoding a protein of 353 amino acids. The CIEBPa amino acid sequences of the Hanwoo show strong conservation with the corresponding sequences reported in other species. The distribution of C/EBP$\alpha$ mRNA in various tissues of Hanwoo aged 12 months were investigated using Northern blotting analysis. The highest expression was detected in adipose tissue and more lower expression was detected in colon and lung. We also identified expression of C/EBPa mRNA in Hanwoo sirloin and adipose tissue aged 12, 26, and 30 months by real-time RT-PCR. The higest expression were detected at 26 months in the sirloin and at 12 and 26 months in the adipose tissue.

Role of $NF-_{{\kappa}B}$ Binding Sites in the Regulation of Inducible Nitric Oxide Synthase by Tyrosine Kinase

  • Ryu, Young-Sue;Hong, Jang-Hee;Lim, Jong-Ho;Bae, So-Hyun;Ahn, Ihn-Sub;Seok, Jeong-Ho;Lee, Jae-Heun;Hur, Gang-Min
    • The Korean Journal of Physiology and Pharmacology
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    • v.5 no.1
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    • pp.55-63
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    • 2001
  • In macrophages, lipopolysaccharide (LPS) alone or in combination with $interferon-{\gamma}\;(IFN-{\gamma})$ has been shown to release a nitric oxide (NO) through the increase of the transcription of the inducible nitric oxide synthase (iNOS) gene. To investigate the exact intracellular signaling pathway of the regulation of iNOS gene transcription by LPS plus $IFN-{\gamma},$ the effects of protein tyrosine kinase (PTK) inhibitor and protein kinase C (PKC) inhibitors on NO production, iNOS mRNA expression, nuclear $factor-_{\kappa}B\;(NF-_{\kappa}B)$ binding activity and the promoter activity of iNOS gene containing two $NF-_{\kappa}B$ sites have been examined in a mouse macrophage RAW 264.7 cells. LPS or $IFN-{\gamma}$ stimulated NO production, and their effect was enhanced synergistically by mixture of LPS and $IFN-{\gamma}.$ The PTK inhibitor such as tyrphostin reduced LPS plus $IFN-{\gamma}-induced$ NO production, iNOS mRNA expression and $NF-_{\kappa}B$ binding activity. In contrast, PKC inhibitors such as H-7, Ro-318220 and staurosporine did not show any effect on them. In addition, transfection of RAW 264.7 cells with iNOS promoter linked to a CAT reporter gene revealed that tyrphostin inhibited the iNOS promoter activity through the $NF-_{\kappa}B$ binding site, whereas PKC inhibitors did not. Taken together, these suggest that PTK, but not PKC pathway, is involved in the regulation of the iNOS gene transcription through the $NF-_{\kappa}B$ sites of iNOS promoter in RAW 264.7 macrophages by LPS plus $IFN-{\gamma}$.

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An observation on the contents of nutrient, fatty acid and changes of lipid peroxide in different storaged commercial pet foods for dogs and cats (애완동물용 식품의 영양성분 및 지방산의 분석과 보관상태별 지질과산화물량의 변화 관찰)

  • Aoki, Hisataka;Sakai, Takeo;Lee, Won-Chang
    • Korean Journal of Veterinary Research
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    • v.37 no.2
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    • pp.439-443
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    • 1997
  • While the contents of protein, fat, fiber, sodium, vitamin A, and vitamin D in dry dog food were 22.0%, 7.8%. 5.4%, 0.36%, 2035 IU/100g and 201 IU/100g, respectevely, those in dry cat food were 29.8%, 6.5%, 4.5%, 0.38%, 1543 IU/100g and 163 IU/100g, respectively. Concerning the lipid contents of pet food, the concentration of unsturated fatty acid was slightly higher of dry dog food(59.2%) than that of dry cat food(55.9%), but the differences was statistically not significant. The lipid peroxide concentration in dry dog food after storage in a refrigerator at $4^{\circ}C$ or dark place of room temperture at $20{\pm}2^{\circ}C$ for 30 days(8.0 nmol/g and 7.8 nmol/g) was not different from the value at the beginning of storage. However, upon storage in direct sunlight at $20{\pm}2^{\circ}C$ of room temperture, the concentration increased 4.9-fold(35.5 nmol/g) after 5 days and 10.2-fold(73.6 nmol/g) after 30 days. These results suggest that pet food may be refrigerated or stored in a dark place after unsealing.

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Extracts from Gracilaria vermiculophylla Prevent Cellular Senescence and Improve Differentiation Potential in Replicatively Senescent Human Bone Marrow Mesenchymal Stem Cells (홍조류인 Gracilaria vermiculophylla 추출물에 의한 노화 골수유래 중간엽줄기세포의 항노화 및 분화능력 개선 효과)

  • Jeong, Sin-Gu;Cho, Tae Oh;Cho, Goang-Won
    • Journal of Life Science
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    • v.28 no.9
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    • pp.1042-1047
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    • 2018
  • The red algae Gracilaria vermiculophylla is widespread on seashores worldwide and has been used as food in Asian countries. Previous studies have reported that extracts of Gracilaria red algae have beneficial anti-oxidant and anti-inflammatory effects. The present study examined the anti-senescence effects of Gracilaria vermiculophylla extracts (GV-Ex) in replicatively senescent human bone marrow mesenchymal stem cells (hBM-MSCs). GV-Ex pretreatment improved the cellular viability of hBM-MSCs that had been injured by oxidative stress. These effects of GV-Ex were confirmed by MTT assay and immunoblot analysis using the apoptotic proteins p53 and cleaved caspase-3. The reactive oxygen species (ROS) levels were examined in long-term cultured Passages 17 (P-17) mesenchymal stem cells (MSC) and compared to P-7 MSC. The ROS accumulation was greater in the P-17 than in the P-7. However, these increased ROS levels in the P-17 were decreased significantly after treatment with GV-Ex, and restoration of the levels of the anti-oxidant enzymes SOD1, SOD2, and CAT was also observed under these conditions. In addition, P-17 hBM-MSC treated with GV-Ex had decreased levels of the senescence proteins p53, p21, and p16. The results show that the ability of P-17 hBM-MSC to differentiate into osteocytes and adipocytes was improved by GV-Ex treatment, suggesting that GV-Ex ameliorates the functional decline of senescent stem cells.

Cloning, Expression, Purification, and Properties of an Endoglucanase Gene (Glycosyl Hydrolase Family 12) from Aspergillus niger VTCC-F021 in Pichia pastoris

  • Pham, Thi Hoa;Quyen, Dinh Thi;Nghiem, Ngoc Minh;Vu, Thu Doan
    • Journal of Microbiology and Biotechnology
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    • v.21 no.10
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    • pp.1012-1020
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    • 2011
  • A gene coding for an endoglucanase (EglA), of the glycosyl hydrolase family 12 and derived from Aspergillus niger VTCC-F021, was cloned and sequenced. The cDNA sequence, 717 bp, and its putative endoglucanase, a 238 aa protein with a predicted molecular mass of 26 kDa and a pI of 4.35, exhibited 98.3-98.7% and 98.3-98.6% identities, respectively, with cDNA sequences and their corresponding endoglucanases from Aspergillus niger strains from the GenBank. The cDNA was overexpressed in Pichia pastoris GS115 under the control of an AOX1 promoter with a level of 1.59 U/ml culture supernatant, after 72 h of growth in a YP medium induced with 1% (v/v) of methanol. The molecular mass of the purified EglA, determined by SDS-PAGE, was 33 kDa, with a specific activity of 100.16 and 19.91 U/mg toward 1% (w/v) of ${\beta}$-glucan and CMC, respectively. Optimal enzymatic activity was noted at a temperature of $55^{\circ}C$ and a pH of 5. The recombinant EglA (rEglA) was stable over a temperature range of $30-37^{\circ}C$ and at pH range of 3.5-4.5. Metal ions, detergents, and solvents tested indicated a slightly inhibitory effect on rEglA activity. Kinetic constants ($K_m$, $V_{max}$, $k_{cat}$, and $k_{cat}/K_m$) determined for rEglA with ${\beta}$-glucan as a substrate were 4.04 mg/ml, 102.04 U/mg, 2,040.82 $min^{-1}$, and 505.05, whereas they were 10.17 mg/ml, 28.99 U/mg, 571.71 $min^{-1}$, and 57.01 with CMC as a substrate, respectively. The results thus indicate that the rEglA obtained in this study is highly specific toward ${\beta}$-glucan. The biochemical properties of rEglA make it highly valuable for downstream biotechnological applications, including potential use as a feed enzyme.