• Title/Summary/Keyword: heat shock protein 40

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Expression and Accumulation of LMW HSPs under Various Heat Shock Conditions (다양한 열처리 조건에서 LMW HSPs의 발현 및 축적량 조사)

  • Kim, Ki-Yong;Jang, Yo-Soon;Lee, Byung-Hyun;Jo, Jinki
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.18 no.4
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    • pp.303-310
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    • 1998
  • We studied expression patterns of thermotolerance gene (BcHSP17.6) in cabbages which was isolated from Chinese cabbage and we will attempt transformation of forage crops with the gene in order to increase thermotolerance of forage crops. Antiserum against a BcHSP17.6 protein was reacted with its antigen. With this antiserum, the accumulation of the 15- to 18-kD LMW HSPs under various heat shock (HS) conditions was quantified. The LMW HSPs began to be detectable at $35^{\circ}C$, and after 4 hours at $40^{\circ}C$ they were accumulated to a maximum level of 1.56 micrograms per 100 micrograms of total proteins in cabbage leaves and remained almost unchanged up to 24 hours after HS. Accumulation of the HSPs was reduced at temperatures higher than $40^{\circ}C$. We conclude that accumulation of these LMW HSPs are necessary for Chinese cabbages to survive at an otherwise lethal temperature.

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Seasons affect the phosphorylation of pork sarcoplasmic proteins related to meat quality

  • Zeng, Xianming;Li, Xiao;Li, Chunbao
    • Animal Bioscience
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    • v.35 no.1
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    • pp.96-104
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    • 2022
  • Objective: Sarcoplasmic proteins include proteins that play critical roles in biological processes of living organisms. How seasons influence biological processes and meat quality of postmortem muscles through the regulation of protein phosphorylation remain to be investigated. In this study, the phosphorylation of sarcoplasmic proteins in pork longissimus muscle was investigated in four seasons. Methods: Sarcoplasmic proteins were extracted from 40 pork carcasses (10 for each season) and analyzed through ProQ Diamond staining for phosphorylation labeling and Sypro Ruby staining for total protein labeling. The pH of muscle, contents of glycogen and ATP were measured at 45 min, 3 h, and 9 h postmortem and the water (P2b, P21, and P22) was measured at 3 h and 9 h. Results: A total of 21 bands were detected. Band 8 (heat shock cognate 71 kDa protein; heat shock 70 kDa protein 1B) had higher phosphorylation level in summer than that in other seasons at 45 min postmortem. The phosphorylation levels of 3 Bands were significantly different between fast and normal pH decline groups (p<0.05). The phosphorylation levels of 4 bands showed negative associations with immobilized water (P21) and positive association with free water (P22). Conclusion: The phosphorylation levels of sarcoplasmic proteins involved in energy metabolism and heat stress response at early postmortem time differed depending on the seasons. These proteins include heat shock protein 70, pyruvate kinase, phosphoglucomutase-1, glucose-6-phosphate isomerase, and carbonic anhydrase 3. High temperatures in summer might result in the phosphorylation of those proteins, leading to pH decline and low water holding capacity.

Changes of Plasminogen Activator Activity under Heat Stress Condition in Porcine Endometrium

  • Hwangbo, Yong;Cheong, Hee-Tae;Park, Choon-Keun
    • Journal of Animal Reproduction and Biotechnology
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    • v.34 no.3
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    • pp.240-246
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    • 2019
  • The aim of this study was to investigate effect of heat stress on expression levels of plasminogen activators (PAs) related mRNAs and proteins, and changes of PAs activity in porcine endometrial explants. The endometrial explants (200 ± 50 mg) were isolated from middle part of uterine horn at follicular phase (Day 19-21) and were pre-incubated in serum-free culture medium at 38.5℃ in 5% CO2 for 18 h. Then, the tissues were transferred into fresh medium and were cultured at different temperature (38.5, 39.5, 40.5 or 41.5℃) for 24 h. The expression level of urokinase-type PA (uPA), type-1 PA inhibitor (PAI-1), type-2 PAI (PAI-2), and heat shock protein-90 (HSP-90) mRNA were analysis by reverse-transcription PCR and proteins were measured by western blotting. The supernatant were used for measurement of PAs activity. In results, mRNA and protein levels of HSP-90 was higher in 41.5℃ treatment groups than other treatment groups (p < 0.05). The expression of uPA, PAI-1, and PAI-2 mRNA were slightly increased by heat stress, however, there were no significant difference. Heat stress condition suppressed expression of active uPA and PAI-2 proteins (p < 0.05), whereas PAI-1 protein was increased (p < 0.01). Although PAI-1 protein was increased and active uPA was decreased, PAs activity was greatly enhanced by exposure of heat stress (p < 0.05). These results suggest that heat stress condition could change intrauterine microenvironment through regulation of PAs activity and other factors regarding with activation of PAs might be regulate by heat stress. Therefore, more studies regarding with regulatory mechanism of PAs activation are needed.

Induced expression of three heat shock proteins mediated by thermal stress in Heortia vitessoides (Lepidoptera: Crambidae)

  • CHENG, Jie;WANG, Chun-Yan;LYU, Zi-Hao;LIN, Tong
    • Entomological Research
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    • v.48 no.5
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    • pp.416-428
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    • 2018
  • To gain an insight into the function of heat shock proteins (HSPs) in insects during thermal stress, three HSP cDNAs were identified in the transcriptome of adult Heortia vitessoides, one of the most destructive defoliating pests in Aquilaria sinensis (Loureiro) Sprenger forests. The open reading frames of HvHsp60, HvHsp70, and HvHsp90 were 1,719, 2,070, and 2,151 bp in length, respectively, and encoded proteins with molecular weights of 61.05, 75.02, and 82.23 kDa, respectively. Sequence analysis revealed that all three HSPs were highly conserved in structure. Regarding the stage-specific expression profiles, HvHsp60, HvHsp70, and HvHsp90 mRNAs were detected in all developmental stages. Regarding the tissue-specific expression profiles, the expression levels of the three HSP genes were different in various larval and adult tissues. Moreover, the expression patterns of heat-stressed larvae, pupae, and adults indicated that HvHsp60, HvHsp70, and HvHsp90 were heat-inducible. In particular, HvHsp60 transcripts increased dramatically in larvae and pupae that were heat-stressed at $40^{\circ}C$ and were upregulated in adults that were heat-stressed at $35^{\circ}C$ and $40^{\circ}C$. The expression of HvHsp70 significantly increased in all of the three different developmental stages at $35^{\circ}C$, $40^{\circ}C$, and $45^{\circ}C$. The expression of HvHsp90 obviously increased at $30^{\circ}C$, $35^{\circ}C$, and $40^{\circ}C$ in larvae and could be induced at $35^{\circ}C$ in pupae and adults. The results suggest that HSP60, HSP70, and HSP90 play a major role in protecting H. vitessoides against high-temperature stress.

The Homologous Region 3 from Bombyx mori Nucleopolyhedrovirus Enhancing the Transcriptional Activity of Drosophila hsp70 Promoter

  • Tang, Shun-Ming;Yi, Yong-Zhu;Zhou, Ya-Jing;Zhang, Zhi-Fang;Li, Yi-Ren;He, Jia-Lu
    • International Journal of Industrial Entomology and Biomaterials
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    • v.9 no.2
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    • pp.235-239
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    • 2004
  • Drosophila melanogaster heat shock protein 70 gene promoter (Dhsp70p) is widely used in transgenic insect to drive exogenous gene, and the homologous region 3 from Bombyx mori nucleopolyhedrovirus (BmNPVhr3) functions as an enhancer for several promoters. To test whether BmNPVhr3 can enhance the Dhsp70ps transcriptional activity, the reporter plasmids, which contain the Dhsp70p, the reporter $\beta$-galactosidase gene with SV40 terminator and BmNPVhr3 fragment, are constructed and transfected into the insect cell lines (Bm-N cells and Sf-21 cells) by lipofectin-mediated method. The results from the transient expression assay show that BmNPVhr3 significantly increases transcriptional activity of Dhsp70p both under the normal condition and under the heat-shock treatment, although the effects are significantly different between in Bm-N cells and in sf-21 cells. The enhancing behavior of BmNPVhr3 on the Dhsp70p is in an orientation-independent manner. Meanwhile, the effects of heat-shock treatment on Dhsp70p alone or Dhsp70p/BmNPVhr3 combination present no significant difference, indicating that BmNPVhr3 only enhances the transcriptional activity of Dhsp70p, but cant alter its characteristic of the response to the heat-shock stress. The above results suggest that the Dhsp70p/BmNPVhr3 combination is more effective one to drive exogenous gene for transgene or stable cell expression system in insects.

Korean ginseng extract ameliorates abnormal immune response through the regulation of inflammatory constituents in Sprague Dawley rat subjected to environmental heat stress

  • Song, Ji-Hyeon;Kim, Kui-Jin;Choi, Seo-Yun;Koh, Eun-Jeong;Park, JongDae;Lee, Boo-Yong
    • Journal of Ginseng Research
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    • v.43 no.2
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    • pp.252-260
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    • 2019
  • Background: Increases in the average global temperature cause heat stress-induced disorders by disrupting homeostasis. Excessive heat stress triggers an imbalance in the immune system; thus protection against heat stress is important to maintain immune homeostasis. Korean ginseng (Panax ginseng Meyer) has been used as a herbal medicine and displays beneficial biological properties. Methods: We investigated the protective effects of Korean ginseng extracts (KGEs) against heat stress in a rat model. Following acclimatization for 1 week, rats were housed at room temperature for 2 weeks and then exposed to heat stress ($40^{\circ}C$/2 h/day) for 4 weeks. Rats were treated with three KGEs from the beginning of the second week to the end of the experiment. Results: Heat stress dramatically increased secretion of inflammatory factors, and this was significantly reduced in the KGE-treated groups. Levels of inflammatory factors such as heat shock protein 70, interleukin 6, inducible nitric oxide synthase, and tumor necrosis factor-alpha were increased in the spleen and muscle upon heat stress. KGEs inhibited these increases by down-regulating heat shock protein 70 and the associated nuclear $factor-{\kappa}B$ and mitogen-activated protein kinase signaling pathways. Consequently, KGEs suppressed activation of T-cells and B-cells. Conclusion: KGEs suppress the immune response upon heat stress and decrease the production of inflammatory cytokines in muscle and spleen. We suggest that KGEs protect against heat stress by inhibiting inflammation and maintaining immune homeostasis.

Classification of Biological Effect of 1,763 MHz Radiofrequency Radiation Based on Gene Expression Profiles

  • Im, Chang-Nim;Kim, Eun-Hye;Park, Ae-Kyung;Park, Woong-Yang
    • Genomics & Informatics
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    • v.8 no.1
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    • pp.34-40
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    • 2010
  • Radiofrequency (RF) radiation might induce the transcription of a certain set of genes as other physical stresses like ionizing radiation and UV. To observe transcriptional changes upon RF radiation, we exposed WI-38, human lung fibroblast cell to 1763 MHz of mobile phone RF radiation at 60 W/kg of specific absorption rate (SAR) for 24h with or without heat control. There were no significant changes in cell numbers and morphology after exposure to RF radiation. Using quantitative RT-PCR, we checked the expression of three heat shock protein (HSP) (HSPA1A, HSPA6 and HSP105) and seven stress-related genes (TNFRSF11B, FGF2, TGFB2, ITGA2, BRIP1, EXO1, and MCM10) in RF only and RF/HS groups of RF-exposed cells. The expressions of three heat shock proteins and seven stress-related genes were selectively changed only in RF/HS groups. Based on the expression of ten genes, we could classify thermal and non-thermal effect of RF-exposure, which genes can be used as biomarkers for RF radiation exposure.

Effect of Environmental Stress on Morphological Change of an Extremely Cadmium-Tolerant Yeast, Hansenula anomala B-7

  • Huh, Nam-Eung;Choi, Nack-Shick
    • Journal of Microbiology and Biotechnology
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    • v.9 no.1
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    • pp.70-77
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    • 1999
  • An extremely cadmium-tolerant budding yeast, Hansenula anomala B-7 underwent a morphological switch in response to either heat shock treatment or cadmium stress, respectively. It exhibited a morphological transition from a unicellular yeast form to a pseudohyphae-like coagulation when subjected to prolonged heat shock treatment. In contrast, the yeast cells showed an irregularity in surface morphology when given thermal stress for a short time. Patterns of proteins expressed in the pseudohyphae-like cells demonstrated that several proteins were overexpressed while others were underexpressed in comparison with those prepared from the cells in the yeast form. It was a striking feature, however, that nearly 40% of the proteins extracted from the cells in the pseudohyphae form appeared to be composed of a single polypeptide. This polypeptide was apparently overexpressed during the pseudohyphae phase and its molecular weight was estimated to be 58 kDa according to SDS-PAGE analysis. However, a significant level of the protein was not observed in the cells before transition to pseudohyphae. The architecture of the cell shape was also damaged when incubated in a medium containing more than 1,000 ppm (8.9mM) of cadmium ions, although able to proliferate at a slow rate. However, the irregularity in the cell morphology exerted either by the brief heat shock treatment or by the cadmium stress with the high concentrations of the metal ions was not repaired, even though the damaged cells were allowed to grow for sufficient time in fresh, cadmium-free medium.

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The Assessment of Toxicity on organic Sludge Using Acetylcholinesterase, Cytochrome P450, and Hsp70 Extracted from Earthworm (Eisenia fetida) (지렁이에서 추출한 Acetylcholinesterase, Cytochrome P450, and Heat Shock protein 70을 이용한 유기성슬러지 독성 평가)

  • Na, Young-Eun;Bang, Hae-Son;Kim, Myung-Hyun;Kim, Min-Kyoung;Roh, Kee-An;Lee, Jung-Taek;Ahn, Young-Joon;Yoon, Seong-Tak
    • Korean Journal of Soil Science and Fertilizer
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    • v.40 no.4
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    • pp.274-279
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    • 2007
  • The toxicitiy of organic sludge such as municipal sewage sludge (MSS), industrial sewage sludge (ISS), alcohol fermentation processing sludge (AFPS) and leather processing sludge (LPS) were evaluated with three environmental biomarkers as acetylcholinesterase, cytochrome P450, and heat shock protein 70 extracted from earthworm (Eisenia fetida). Their toxicities were compared with those of pig manure compost (PMC). MSS, ISS, LPS, and AFPS did not significantly affect the acetylcolinesterase activity, whereas only the elutriate of PMC slightly was increased the activity. MSS, AFPS, and PMC tended to slightly inhibit the cytochrome $P_{450}$ activity, but ISS and LPS showed significantly the inhibitory effect on cytochrome $P_{450}$. The hsp70 expression began to increase after treatments and showed high induction at 6 hour, followed by zero level at around 12 hour. The quantity of the hsp70 expressed by elutriate treatments of PMC, AFPS, MSS, ISS, and LPS was 1.9, 3.0, 3.3, 4.4, and 4.7 fold higher than that of distilled water. These results indicate that in toxicity tests of five organic waste materials, four kinds of sludge materials appeared more toxic than PMC. Results of AChE, P450, and hsp70 of earthworm might be useful for expecting or assessing an effect by exposure of organic wastes to earthworms in soil.

Fluctuation of Temperature Induces Pathogenicity of Streptococcus iniae and Changes of Immunology Related Genes of Korean Rockfish, Sebastes schlegeli

  • EunYoung Min;Seon-Myeong Jeong;Hyun-Ja Han;Miyoung Cho
    • Korean Journal of Ecology and Environment
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    • v.56 no.4
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    • pp.420-429
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    • 2023
  • This study was designed to examine the immune response in Korean rockfish during water temperature fluctuation and to elucidate the factors contributing to streptococcal pathogenesis in cultured Korean rockfish, S. schlegeli. We investigated cumulative mortality against Streptococcus iniae (FP5228 strain) infection in the exposed Korean rockfish (39.7±5.8 g) to environmentally relevant temperature (Control, 23℃; High temperature, 28℃ and 23℃ and 28℃ with 12 hours interval exchange, 23↔28℃) for 48 hours. Also, the expression of the mRNA related to the immune response genes (heat shock protein 70, interleukin1β, lysozyme g-type and thioredoxin-like 1) were measured in spleen and head kidney by real-time PCR analysis in the exposed fish to thermal stress. In this study, the combined stress with bacterial challenge in fishes exposed to thermal stress lowered the survival rate than that of control (23℃). The cumulative mortality in the group of control, 28℃ and 23↔28℃ was 24%, 24% and 40% (P<0.05), respectively. Also, thermal stress modulated the mRNA level of immune related genes; heat shock protein 70, interleukin-1β, lysozyme g-type and thioredoxin-like 1 in Korean rockfish. The present study indicates that a high and sudden water temperature change affect immune responses and reduce the disease resistance in Korean rockfish.