• Title/Summary/Keyword: heat-shock stress

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Expression of HSP70 mRNA and Protein based on the Thermal Stress in the Primary Hepatocyte Culture of Walleye Pollock (Gadus chalcogrammus) (명태(Gadus chalcogrammus)의 일차 간세포 배양에서 온도 스트레스에 따른 HSP70 mRNA와 단백질 발현)

  • Kim, So-Sun;Lee, Chang-Ju;Park, Jang-Su
    • Journal of Environmental Science International
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    • v.29 no.6
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    • pp.633-641
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    • 2020
  • Water temperature is one of the most important factors of fish survival, affecting the habitat, migration route, development, and reproduction. This experiment studied the induction level of heat shock protein (HSP70) mRNA and protein in a walleye pollock (Gadus chalcogrammus) primary hepatocyte culture based on different temperatures. Hepatocytes were attached at 7.5℃ for 24 hours. Hsp70 induction levels were then measured for 48 hours at 5, 8, 11, 14, and 17℃. The induction level was lowest at 5℃ and generally increased with temperature until 14℃. The induction level was reduced at 17℃, indicating that 14℃ is the highest tolerable temperature for hepatocytes. These data indicate that primary hepatocyte cell culture is under no stress at 5 and 8℃. Temperatures greater than 11℃ induce stress, showing similar induction patterns in both mRNA and protein in hepatocytes. The results suggest that 14℃ is the maximum internal defense temperature of walleye pollock survival.

STATE-OF-THE-ART TECHNOLOGY USING GENETICALLY-ENGINEERED BIOLUMINESCENT BACTERIA AS ENVIRONMENTAL BIOSENSORS

  • Gu, Man-Bock
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2000.04a
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    • pp.94-99
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    • 2000
  • Bioluminescence is being used as a prevailing reporter of gene expression in microorganisms and mammalian cells. Bacterial bioluminescence draws special attention from environmental biotechnologists since it has many advantageous characteristics, such as no requirement of extra substractes, highly sensitive, and on-line measurability. Using bacterial bioluminescence as a reporter of toxicity has replaced the classical toxicity monitoring technology of using fish or daphnia with a cutting-edge technology. Fusion of bacterial stress promoters, which control the transcription of stress genes corresponding to heat-shock, DNA-, or oxidative-damaging stress, to the bacterial lux operon has resulted in the development of novel toxicity biosensors with a short measurement time, enhanced sensitivity, and ease and convenient usage. Therefore, these recombinant bioluminescent bacteria are expected to induce bacterial bioluminescence when the cells are exposed to stressful conditions, including toxic chemicals. We have used these recombinant bioluminescent bacteria in order to develop toxicity biosensors in a continuous, portable, or in-situ measurement from for air, water, and soil environments. All the data obtained from these toxicity biosensors for these environments were found to be repeatable and reproducible, and the minimum detection level of toxicity was found to be ppb (part per billion) levels for specific chemicals.

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Degradation of the Parasporal Crystal Proteins of Bacillus thuringiensis and Induction of Stress Protein Synthesis in Bt $\delta$-endotoxin Ingested Larvae of Fall Webworm, Hyphantria cunea (Bacillus thuringiensis 의 내독소 단백질의 분해와 흰불나방 섭취유충에서 스트레스 단백질 합성의 유발)

  • 전향미;유용만;강석권;서숙재
    • Korean journal of applied entomology
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    • v.33 no.3
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    • pp.178-183
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    • 1994
  • The prolein components of each crystal toxin of Bacillus thuringiensis kurstaki and alzawai were separated by SDS-PAGE. The major clqistal proteins from both shins were composed of paiypeptides having molecular weights of 130 kd and 64 kd. Digestive mixture of both. toxins with t~ypsin and gut juices shared 62 kd polypeptide which may be major actwe toxh. However, aizawal produced much less amount of 62 kd polypeptide than kurstaki did. On ingestion with Bt &-endotoxin, larvae of Hyphantria cunea developed 45 kd stress protein in the several tissues including fat body, which was induced by heat and cold shock.

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Reproductive Performance of Holstein Dairy Cows Grazing in Dry-summer Subtropical Climatic Conditions: Effect of Heat Stress and Heat Shock on Meiotic Competence and In vitro Fertilization

  • Pavani, Krishna;Carvalhais, Isabel;Faheem, Marwa;Chaveiro, Antonio;Reis, Francisco Vieira;da Silva, Fernando Moreira
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.3
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    • pp.334-342
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    • 2015
  • The present study was designed to evaluate how environmental factors in a dry-summer subtropical climate in Terceira-Azores (situated in the North Atlantic Ocean: $38^{\circ}43^{\prime}N27^{\circ}12^{\prime}W$) can affect dairy cow (Holstein) fertility, as well as seasonal influence on in vitro oocytes maturation and embryos development. Impact of heat shock (HS) effects on in vitro oocyte's maturation and further embryo development after in vitro fertilization (IVF) was also evaluated. For such purpose the result of the first artificial insemination (AI) performed 60 to 90 days after calving of 6,300 cows were recorded for one year. In parallel, climatic data was obtained at different elevation points (n = 5) from 0 to 1,000 m and grazing points from 0 to 500 m, in Terceira island, and the temperature humidity index (THI) was calculated. For in vitro experiments, oocytes (n = 706) were collected weekly during all year, for meiotic maturation and IVF. Further, to evaluate HS effect, 891 oocytes were collected in the cold moths (December, January, February and March) and divided in three groups treated to HS for 24 h during in vitro maturation at: C (Control = $38.5^{\circ}C$), HS1 ($39.5^{\circ}C$) and HS2 ($40.5^{\circ}C$). Oocytes from each group were used for meiotic assessment and IVF. Cleavage, morula and blastocyst development were evaluated respectively on day 2, 6, and 9 after IVF. A negative correlation between cow's conception rate (CR) and THI in grazing points (-91.3%; p<0.001) was observed. Mean THI in warmer months (June, July, August and September) was $71.7{\pm}0.7$ and the CR ($40.2{\pm}1.5%$) while in cold months THI was $62.8{\pm}0.2$ and CR was $63.8{\pm}0.4%$. A similar impact was obtained with in vitro results in which nuclear maturation rate (NMR) ranged from 78.4% (${\pm}8.0$) to 44.3% (${\pm}8.1$), while embryos development ranged from 53.8% (${\pm}5.8$) to 36.3% (${\pm}3.3$) in cold and warmer months respectively. In vitro HS results showed a significant decline (p<0.05) on NMR of oocytes for every $1^{\circ}C$ rising temperature ($78.4{\pm}8.0$, $21.7{\pm}3.1$ and $8.9{\pm}2.2$, respectively for C, HS1, and HS2). Similar results were observed in cleavage rate and embryo development, showing a clear correlation (96.9 p<0.05) between NMR and embryo development with respect to temperatures. Results clearly demonstrated that, up to a THI of 70.6, a decrease in the CR occurs in first AI after calving; this impairment was confirmed with in vitro results.

A Comparative Experiment on Thermal Stress Failure of Vacuum Glazing applied in Curtain Wall at Spandrel area (커튼월 스팬드럴용 진공유리의 열파손에 대한 비교실험)

  • Kim, Seung-Chul;Yoon, Jong-Ho;Shin, U-Cheol;Ahn, Jung-Hyuk
    • KIEAE Journal
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    • v.16 no.3
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    • pp.121-128
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    • 2016
  • Purpose: The vacuum glazing should constantly retain the gap in vacuum state to maintain high thermal performance. To do so, pillars are used to prevent the glazing from clinging to each other by the atmospheric pressure and therefore surface of the vacuum glazing is consistently affected by residual stress. The vacuum glazing could be applied to curtain wall systems at spandrel area to fulfill a rigorous domestic standard on U-value of the external wall. However, this can lead to high glazing temperature increase by heat concentration at a back panel and finally thermal stress breakage. This study experimentally determined weakness of the vacuum glazing systems on the thermal stress breakage and investigated effect of the residual stress. Method: The experiment first built two scale-down mock-up facilities that replicate the spandrel area in curtain wall, and then installed single low-e glass and vacuum glazing respectively. The two mock-up facilities were exposed to outside to induce the thermal stress breakage. Result: The experiment showed that the temperature occurred the thermal stress breakage was $114.4^{\circ}C$ for the single low-e glass and $118.9^{\circ}C$ for the vacuum glazing respectively. The result also showed the vacuum glazing reached the critical point earlier than the single low-e glass, which means that the vacuum glazing has high potential to occur the thermal shock breakage. In addition, the small temperature difference between two glazing indicates that the residual stress scarcely affects breakage of the vacuum glazing.

Overexpression of NtHSP70-1 Protects Chlorophyll from High Temperature in Plants (NtHSP70-1에 의한 클로로필의 고온 내성 효과)

  • Cho, Eun-Kyung;Hong, Choo-Bong
    • Journal of Life Science
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    • v.18 no.3
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    • pp.304-310
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    • 2008
  • Heat shock protein 70 (HSP70) is known as molecular chaperone, the fundamental protein participating in various processes, from nascent protein synthesis to protection of proteins during abiotic stresses and developmental programs. However, their biological functions in plants are not yet well known. Here, NtHSP70-1 (AY372069), HSP70 of Nicotiana tabacum induced by heat stress was investigated. To analyze the protective role of NtHSP70-1, transgenic tobacco plants, which constitutively overexpressed NtHSP70-1 as well as contained either the vector alone or having NtHSP70-1 in the antisense orientation, were constructed. The altered NtHSP70-1 levels in plants were confirmed by western blotting and transgenic sense lines exhibited tolerance to heat stress. Seedlings with the constitutively expressed NtHSP70-1 grew as green or healthy plants after heat stress. In contrast, transgenic vector or antisense lines exhibited yellowing of leaves or some delay in growth, which finally led to death. Evaluation of chlorophyll contents of heat-shocked transgenic tobacco seedlings indicated that NtHSP70-1 contributes to thermotolerance by preventing chlorophyll synthesis in plants.

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.

Expression of Heat Shock Protein in Cytokine Stimulated PDL Cells and Inflamed Gingival Tissue (염증성 치은조직과 치주인대세포에서 Cytokine에 의해 유도되는 열충격단백 발현에 관한 연구)

  • Cho, In-Ho;Kim, Doek-Kyu;Kim, Eun-Cheol;You, Hyung-Keun;Shink, Hyung-Shin
    • Journal of Periodontal and Implant Science
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    • v.28 no.1
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    • pp.103-120
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    • 1998
  • Prokaryotic and eukaryotic cells respond to heat stress and other environmental abuses by synthesizing a small set of stress proteins and by inhibiting post-transcription synthesis of normal proteins. The purpose of the present study was to document the stress response produced by inflamed gingival tissue in vivo, and cytokine inducted human periodontal ligament cells. Human PDL cells were exposed to TNF-$\alpha$(1ng/ml), INF-$\gamma$(200 U/ml), LPS(100ug/ml), combination of cytokine, and SDS-PAGE gels running and Western blotting analysis was done. In vivo studies, the healthy gingival tissusse of a control group and inflamed gingival tissue of adult periodontitis were studied by immunohistochemistry and histology. The results were as follows 1. HSP 47 was distributed on basal layer in healthy gingiva, but stronger stained in basal, suprabasal, and spinous layer of inflamed gingiva. 2. HSP 47 was rare on endothelial cells and mononuclear cells in healthy gingiva, but stronger expressed in inflamed gingiva. 3. HSP 70 expression was rare on epihelium and inflammatory cells hi both healthy & inflamed gingiva. 4. HSP 70 was actively expressed on endothelial cells and inflammatory cells of capillary lumen in moderately & mild inflamend gingiva. 5. PDL cells showed low level of HSP 47 protein expression which was significantly induced by cytokine stimulation (LSP only and combination). 6. Maximum HSP 70 protein induction was seen with stimulation by a combination of the cytokine, Combination of TNF-$\alpha$, INF-$\gamma$, LPS have been shown to synergistically effects of HSP 70 expression. On the above findings, HSP Is influenced by cytokine and chronic inflammation in vivo, and may be involved in protection of tissue during periodontal inflammatiom.

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Bag-1L is a Stress-withstand Molecule Prevents the Downregulation of Mcl-1 and c-Raf Under Control of Heat Shock Proteins in Cisplatin Treated HeLa Cervix Cancer Cells

  • Ozfiliz, Pelin;Arisan, Elif Damla;Coker-Gurkan, Ajda;Obakan, Pinar;Eralp, Tugce Nur;Dinler-Doganay, Gizem;Palavan-Unsal, Narcin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.11
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    • pp.4475-4482
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    • 2014
  • Background: Cisplatin, a DNA damaging agent, induces apoptosis through increasing DNA fragmentation. However, identification of intrinsic resistance molecules against Cisplatin is vital to estimate the success of therapy. Bag-1 (Bcl-2-associated anthanogene) is one anti-apoptotic protein involved in drug resistance impacting on therapeutic efficiency. Elevated levels of this protein are related with increase cell proliferation rates, motility and also cancer development. For this reason, we aimed to understand the role of Bag-1 expression in Cisplatin-induced apoptosis in HeLa cervix cancer cells. Cisplatin decreased cell viability in time- and dose-dependent manner in wt and Bag-1L+HeLa cells. Although, $10{\mu}M$ Cisplatin treatment induced cell death within 24h by activating caspases in wt cells, Bag-1L stable transfection protected cells against Cisplatin treatment. To assess the potential protective role of Bag-1, we first checked the expression profile of interacting anti-apoptotic partners of Bag-1. We found that forced Bag-1L expression prevented Cisplatin-induced apoptosis through acting on Mcl-1 expression, which was reduced after Cisplatin treatment in wt HeLa cells. This mechanism was also supported by the regulation of heat shock protein (Hsp) family members, Hsp90 and Hsp40, which were involved in the regulation Bag-1 interactome including several anti-apoptotic Bcl-2 family members and c-Raf.

Expression of Cu/Zn SOD Protein Is Suppressed in hsp 70.1 Knockout Mice

  • Choi, S-Mi;Park, Kyung-Ae;Lee, Hee-Joo;Park, Myoung-Sook;Lee, Joung-Hee;Park, Kyoung-Chan;Kim, Man-Ho;Lee, Seung-Hoon;Seo, Jeong-Sun;Yoon, Byung-Woo
    • BMB Reports
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    • v.38 no.1
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    • pp.111-114
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    • 2005
  • Heat shock proteins (HSPs) are known to protect cells from oxidative stress and other types of injuries. We previously reported the neuroprotective effect of HSP70 following cerebral ischemia and reperfusion using hsp 70.1 knockout (KO) mice. However, the precise role of HSP70 in neuroprotection has not been established yet. The purpose of this study was to investigate the relationship between HSP70 and antioxidant enzymes using hsp 70.1 KO mice. The activities of both SOD-1 and SOD-2 were significantly decreased in hsp 70.1 KO mice than in the wild type (WT) littermates. SOD-1 protein level in the hsp 70.1 KO mice was lower than that of WT. We speculate that HSP70 might be involved in regulation of expression of SOD-1 at the level of transcription or by post-transcriptional modification.