• 제목/요약/키워드: Heat Shock Protein

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Comparison of Stress Indicators in Blood and Muscle of Pigs in Conventional and Animal Welfare Farms (일반 및 동물복지농장 돼지의 혈액 및 근육 내 스트레스 인자 비교)

  • Lee, Jeong-Eun;Park, Jin-Ryong;Kang, Da-Rae;Kim, Hee-Eun;Nam, Ki-Chang;Shim, Kwan-Seob
    • Korean Journal of Organic Agriculture
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    • v.28 no.4
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    • pp.627-641
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    • 2020
  • Intensive farming methods that do not guarantee animal welfare can induce stress in pigs. Stress, in turn, can reduce their disease resistance and influence their hormones and metabolites in such a manner that productivity is negatively affected. This study was conducted to compare the stress related factors and blood characteristics of pigs raised on conventional farms and those raised on animal welfare farms. We measured the levels of cortisol, epinephrine and norepinephrine, biochemical parameters in blood and glycogen, L-lactate and heat shock protein 70 (HSP70) in muscle, as physiological markers of indicating the stress in conventional farm pigs (Control, n=10) and animal welfare farm pigs (Welfare, n=10). We found that there was a significant difference in the albumin-globulin ratio (A/G ratio), as well as the albumin (ALB), blood urea nitrogen (BUN) and aspartate aminotransferase (AST) levels between the two farms. Epinephrine was significantly higher in conventional farm, while level of norepinephrine was higher in animal welfare farm. There was no significant difference in cortisol, which is known as a stress hormone, across the two groups of farms. Muscular glycogen content was significantly high in animal welfare farm pigs. While L-lactate tended to be low in the animal welfare farm pigs, the difference between them and the conventional farm cohorts was not significant. HSP70 showed high levels of expression in conventional farm. Thus, we suggested that blood parameter results showed a stress response in the livers of conventional farm, and that catecholamine hormones, glycogen, L-lactate and HSP70 can be used as physiological factors of assessing animal welfare.

Characterization of Gel16 as a Cytochrome P450 in Geldanamycin Biosynthesis and in-silico Analysis for an Endogenous Electron Transport System

  • Rimal, Hemraj;Yu, Sang-Cheol;Lee, Byeongsan;Hong, Young-Soo;Oh, Tae-Jin
    • Journal of Microbiology and Biotechnology
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    • v.29 no.1
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    • pp.44-54
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    • 2019
  • Geldanamycin and its derivatives, inhibitors of heat shock protein 90, are considered potent anticancer drugs, although their biosynthetic pathways have not yet been fully elucidated. The key step of conversion of 4,5-dihydrogeldanamycin to geldanamycin was expected to catalyze by a P450 monooxygenase, Gel16. The adequate bioconversions by cytochrome P450 mostly rely upon its interaction with redox partners. Several ferredoxin and ferredoxin reductases are available in the genome of certain organisms, but only a few suitable partners can operate in full efficiency. In this study, we have expressed cytochrome P450 gel16 in Escherichia coli and performed an in vitro assay using 4,5-dihydrogeldanamycin as a substrate. We demonstrated that the in silico method can be applicable for the efficient mining of convenient endogenous redox partners (9 ferredoxins and 6 ferredoxin reductases) against CYP Gel16 from Streptomyces hygroscopicus. The distances for ligand FDX4-FDR6 were found to be $9.384{\AA}$. Similarly, the binding energy between Gel16-FDX4 and FDX4-FDR6 were -611.88 kcal/mol and -834.48 kcal/mol, respectively, suggesting the lowest distance and binding energy rather than other redox partners. These findings suggest that the best redox partners of Gel16 could be NADPH ${\rightarrow}$ FDR6 ${\rightarrow}$ FDX4 ${\rightarrow}$ Gel16.

DNA Array Analysis of Changes in Gene Expression Profile in DHEA-induced PCO

  • Yu, Jeong-Min;Yoo, Seong-Jin;Kim, Do-Rim;Youm, Mi-Young, Kim, Jee-Yun;Kang, Sung-Goo
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.112-112
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    • 2002
  • Under normal conditions, women produce a single dominant follicle that participates in a single ovuation each menstrual cycle. But Polycystic ovary syndrome(PCOS) conditions, folliculogenesis does not proceed normally. This condition leads to the accumlation of large numbers of small graffian follicles in which the theca interstitial cells (TIC) produce abnormally large amounts of androgen. PCOS is probably the most common endocrine disorder, affecting women of reprodutive age with 5-10% prevalence estimate. Chronic anovulation, hyperandrogenism, hirsutism, obesity, infertility and polycystic ovaries are clinical hallmarks of women with PCOS. Its etiology remains unknown. To investigate the gene expression pattern of ovary in PCO-induced rat, we used cDNA expression analysis. Total RNA was extracted from the ovary of PCO-induced rat and reverse-transcribed in the presence of[$\alpha$$^{32}$P]-dATP Which were hybridized to Atlas$^{TM}$ Rat Toxicology 1.2 array (Clontech) representing approximately 1176 rat genes. We compared gene expression between ovary of pco-induced immature female rats and control. Differential gene expression profiles were revealed (LIFR-alpha, ADRA1A, Heat shock 90-kDa protein A, PDGFRA). Reverse transcription-polymerase chain reaction(RT-PCR) was used to validate the relative expression pattern obtained by the cDNA array. The precise relationship between the altered expression of genes and PCO is a matter of further investigation. This study was supported by Korea Science and Engineering Foundation(KOSEF)

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Bioaccumulation and Expressions of Stress Response Genes in Benthic Oligochaete Worm Tubifex tubifex to Exposure of Cadmium-spiked Sediment (카드뮴 (Cd) 노출 퇴적물에 따른 실지렁이 Tubifex tubifex의 체내 축적과 스트레스 반응 유전자 발현)

  • Ji-Hoon Kim;Won-Seok Kim;Kiyun Park;Ihn-Sil Kwak
    • Korean Journal of Ecology and Environment
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    • v.56 no.4
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    • pp.320-329
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    • 2023
  • Cadmium (Cd), a heavy metal found in the aquatic environment, accumulates in organisms through the food chain. In the study, we investigated the survival rates, measurement of body Cd levels, and expression analysis of the stress response genes (Heat shock protein 70: HSP70 and Heat shock protein 60: HSP60) and antioxidant enzyme Glutathione S-Transferases (GST) on benthic oligochaete worm Tubifex tubifex exposed three concentrations of Cd, to analyze the bioaccumulation and changes of stress gene expressions to exposure toxicity of the Cd-spiked sediment. Survival rates of T. tubifex exposed to the Cdspiked sediment were 93% at 0.4 mg kg-1 Cd, 96% at 1.87 mg kg-1 Cd, and 93% at 6.09 mg kg-1 Cd for 10 days. Cd concentration in the body of T. tubifex was higher than that in the sediment. After Cd exposures for 10 days, the body Cd levels were 18.4 mg kg-1, 13.06 mg kg-1, and 79.11 mg kg-1 at exposed three concentrations of Cd, respectively. Upregulation of HSP70 gene expression was observed at all concentrations of exposed Cd as a time-dependent manner, whereas transcriptional expression of the HSP60 gene increased as a timedependent manner in T. tubifex exposed to the relative high concentration (6.09 mg kg-1) of Cd. However, GST gene expression increased on day 1 at all concentrations after Cd exposures, and then downregulated until 10 days. These results indicate to ecotoxicological and molecular effects in benthic oligochaete worm T. tubifex to Cd-spiked sediment and provide the basic information for the utilization of environmental toxicity assessment using the T. tubifex as a aquatic pollution indicator species.

Comparison of Biological Responses and Heat Shock Protein 70 Expression in Chironomidae by Exposure UV and O3 (UV와 O3 노출에 따른 깔따구류의 생물학적 반응 및 열충격 단백질 70 발현)

  • Ji-Hoon Kim;Won-Seok Kim;Jae-Won Park;Bong-Soon Ko;Kiyun Park;Ihn-Sil Kwak
    • Korean Journal of Ecology and Environment
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    • v.56 no.4
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    • pp.430-439
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    • 2023
  • UV and O3 are materials used in the water treatment process, and many studies have been reported to remove organic matters, contaminants, and microorganisms. In this study, we were investigated effects of Chirnomidae (Chironomus flaviplumus, Chironomus riparius), which are contamination indicator species to exposure UV and O3 for the survival rate, body color change and gene expression response. The survival rate of C. flaviplumus exposed to UV decreased to about 70% after 24 hours, and C. riparius about 50%. There was no change in the survival rate of C. flaviplumus exposed to O3, and C. riparius decreased to 95% after 10 minutes of exposure, but there was no change during the subsequent exposure time. In addition, UV and O3 exposure to the two species in body color faded in a time-dependent. In the HSP70 gene expression, C. riparius showed an increase in expression after UV exposure compared to the control group, and a significant difference was shown 12 hours after exposure (P<0.05). C. flaviplumus exposed to O3 showed a relatively low expression compared to the control group, and showed a significant difference at 10 minutes and 1 hour after exposure (P<0.05). These results reported the ecotoxicological effects on Chironomidae according to UV and O3 exposure. Therefore, the results of this study can be used as basic data to understand the effects of UV and O3, which are disinfectants used in water treatment plants, on Chirnomidae entering plants.

Oxidative Stress-dependent Structural and Functional Regulation of 2-cysteine Peroxiredoxins In Eukaryotes Including Plant Cells (산화 스트레스에 의존한 식물 및 진핵세포 2-시스테인 퍼록시레독신의 기능 조절)

  • Jang, Ho-Hee;Kim, Sun-Young;Lee, Sang-Yeol
    • Journal of Plant Biotechnology
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    • v.33 no.1
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    • pp.1-9
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    • 2006
  • Peroxiredoxins (Prxs) are ubiquitously distributed and play important functions in diverse cellular signaling systems. The proteins are largely classified into three groups, such as typical 2-Cys Prx, atypical 2-Cys Prx, and 1-Cys Prx, that are distinguished by their catalytic mechanisms and number of Cys residues. From the three classes of Prxs, the typical 2-Cys Prx containing the two-conserved Cys residues at its N-terminus and C-terminus catalyzes $H_2O_2$ with the use of thioredoxin (Trx) as an electron donor. During the catalytic cycle, the N-terminal Cys residue undergoes a peroxide-dependent oxidation to sulfenic acid, which can be further oxidized to sulfinic acid at the presence of high concentrations of $H_2O_2$ and a Trx system containing Trx, Trx reductase, and NADPH. The sulfinic acid form of 2-Cys Prx is reduced by the action of sulfiredoxin which requires ATP as an energy source. Under the strong oxidative or heat shock stress conditions, 2-Cys Prx in eukaryotes rapidly switches its protein structure from low-molecular-weight species to high-molecular-weight protein structures. In accordance with its structural changes, the protein concomitantly triggers functional switching from a peroxidase to a molecular chaperone, which can protect its substrate denaturation from external stress. In addition to its N-terminal active site, the C-terminal domain including 'YF-motif' of 2-Cys Prx plays a critical role in the structural changes. Therefore, the C-terminal truncated 2-Cys Prxs are not able to regulate their protein structures and highly resistant to $H_2O_2$-dependent hyperoxidation, suggesting that the reaction is guided by the peroxidatic Cys residue. Based on the results, it may be concluded that the peroxidatic Cys of 2-Cys Prx acts as an '$H_2O_2$-sensor' in the cells. The oxidative stress-dependent regulation of 2-Cys Prx provides a means of defense systems in cells to adapt stress conditions by activating intracellular defense signaling pathways. Particularly, 2-Cys Prxs in plants are localized in chloroplasts with a dynamic protein structure. The protein undergoes conformational changes again oxidative stress. Depending on a redox-potential of the chloroplasts, the plant 2-Cys Prx forms super-molecular weight protein structures, which attach to the thylakoid membranes in a reversible manner.

Studies on the Biological Action of Hyperthermia on Tumor Cell Mortality (1) (腫瘍細胞가 致死에 미치는 溫熱處理의 生物學的 作用에 관한 硏究(1))

  • Kang, Man-Sik;Lee, Chung-Choo
    • The Korean Journal of Zoology
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    • v.26 no.2
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    • pp.69-81
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    • 1983
  • The biological effect of hyperthermia on the SCK tumor cells in vitro were analyzed in several respects. A comparision of the survival curve of SCK tumor cells in vitro and in vivo following hyperthermia demonstrated that the cytocidal effect of heating is far greater on the cells in vivo than on the cells in vitro. The pH change in the SCK tumor upon being heated at $43.5^\\circC$ started out at 7.05 and increased to 7.18 during the first 7 min of heating and then rapidly declined to 6.67 by 30 min. Contrary to the decrease in pH in the heated tumors, the pH in the muscle increased significantly when heated to $43.5-45.0^\\circC$. Following hyperthermia at $43.5^\\circC$ for 30 min, a maximum increase in the lactic acid content in the tumor and liver was observed at 1 hr and 3 hr, respectively. The increase in the tumor was followed by a gradual decrease below the control level, whereas the increase in the liver was maintained at quite a steady level for 24 hr. The hyperthermia at $43.5^\\circC$ for 1 hr exhibited a general tendency that high molecular proteins decrease markedly, whereas most of low molecular proteins increase. The most prominent change was that the heat shock protein 70K increased significantly along with other low molecular proteins in heat shocked cells.

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Effect of Suboptimal Temperature Incubation on the Resistance of Lactobacillus acidophilus CT 01 to Storage and Drying (저온배양에 따른 Lactobacillus acidophilus CT 01의 저장 및 건조에 대한 저항성)

  • Yu Keun-Hyung;Kwon Il-Kyoung;Kim Gur-Yoo
    • Food Science of Animal Resources
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    • v.25 no.1
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    • pp.92-97
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    • 2005
  • This study was carried out to determine the storage, cryotolerance, heat and drying resistance, when Lactobacillus acidophilus CT 01 isolated from preweaned piglet feces growing at suboptimal temperature. L. acidophilus CT 01 suboptimal temperature incubated for 48 hours had the slowest growth rate at 22℃ but the highest viable cell number after 36 hours at 22℃, with 1.3×10/sup 9/ CFU/mL. In case of 4 and 20℃ storage, the suboptimal temperature incubated groups had a viability higher than the control (p<0.01). The cryotolerance of suboptimal temperature incubated L. acidophilus CT 01 was a higher than the control (p<0.01). When L. acidophilus CT 01 was heat treated at 60℃ for 15 minutes and 30 minutes, the suboptimal temperature incubated L. acidophilus CT 01 at 22℃ had a viability higher more than the control (p<0.01). L. acidophilus CT 01 incubated suboptimal temperature was inoculated by 30% to the carrier, and dried at 50℃ for 12 hours had the highest viability in the suboptimal temperature incubated L. acidophilus CT 01 at 28℃.

Pre-Exercise Protective Effects Against Renal Ischemic Reperfusion Injury in Hsp 70.1 Knockout Mice (Hsp70.1유전자결핍된 마우스에서 허혈 재관류 신장손상에 대한 전처치 운동의 보호효과)

  • Lee, Jin;Kim, Won-Kyu
    • Journal of Life Science
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    • v.20 no.4
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    • pp.555-560
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    • 2010
  • The objective of this study was to investigate levels of serum creatinine, CuSOD and MnSOD protein expression in the kidney after renal ischemic reperfusion with pre-exercise using heat shock protein 70.1 in knock-out mice (KO). The C57/BL6 strain (Wild type: WT) and KO were divided into 4 groups as follows: Sham control group (Sham), pre-exercise group (Ex), pre-exercise +ischemia group (Ex+IR), and ischemia group (IR). CuSOD and MnSOD expression were significantly decreased (p<0.01, p<0.05) and blood creatinine concentration was significantly increased (p<0.01) in the IR group of KO. In contrast, CuSOD and MnSOD expression in the Ex+IR group of KO were higher than the IR group, while creatinine concentration was significantly lower. These results suggest that Hsp70 is directly correlated to renal ischemic reperfusion injury. Pre-exercise in renal ischemia might prevent or inhibit positive oxidative stress inhibitory effects by increasing anti-oxidative enzymes (CuSOD, MnSOD) within the kidney and improve to prevent renal function. Thus, pre-exercise may have a protective role against renal injury after renal ischemia.

Identification of Genes Differentially Expressed by Oryctes rhinoceros nudivirus Infection in the Korean Rhinoceros Beetle, Allomyrina dichotoma (장수풍뎅이(Korean Rhinoceros Beettle, Allomyrina dichotoma)에 Oryctes rhinoceros nudivirus 감염 특이적으로 발현하는 유전자 동정)

  • Kwon, Kisang;Yoo, Bo-Kyung;Suh, Hyun-Woo;Ko, Young Hwa;Kim, Hong Geun;Lee, Seokhyun;Park, Kwan-Ho;Choi, Ji-Young;Kwon, O-Yu
    • Journal of Life Science
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    • v.25 no.8
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    • pp.942-946
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    • 2015
  • The Korean rhinoceros beetle (Allomyrina dichotoma) is popular as a pet and as a food ingredient, and it is commercially distributed in Korea. It is also traditionally regarded as a medicine for liver-related diseases. Recently, the Oryctes rhinoceros nudivirus was introduced from Southeast Asia. This virus is reported as a disease factor for A. dichotoma in mass-rearing facilities, and economic losses due to this viral infection have been increasing in Korea since the 2010s. In this study, we observed serious structural changes in the fat body and the intestine of virus-infected beetles. We report five genes that are up-regulated by the viral infection in the intestine: BTF3H4-like (transcription factor BTF3 homolog 4-like), SPS-like (serine proteinase stubble-like), COPB1 (coatomer protein complex, subunit beta 1), T-CP (T-complex 1 subunit gamma), and HSP70 HSP70 (heat shock protein 70). The results may provide a clue for the early diagnosis and disease-treatment that occurs in mass-rearing facilities. The improvement of stable productivity will increase the farmers’ income, and quality control of beetle-breeding will help industries to utilize this beetle as a promising food ingredient.