• Title/Summary/Keyword: Heat shock protein(HSP)

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Transcriptome-based identification of water-deficit stress responsive genes in the tea plant, Camellia sinensis

  • Tony, Maritim;Samson, Kamunya;Charles, Mwendia;Paul, Mireji;Richard, Muoki;Mark, Wamalwa;Stomeo, Francesca;Sarah, Schaack;Martina, Kyalo;Francis, Wachira
    • Journal of Plant Biotechnology
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    • v.43 no.3
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    • pp.302-310
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    • 2016
  • A study aimed at identifying putative drought responsive genes that confer tolerance to water stress deficit in tea plants was conducted in a 'rain-out shelter' using potted plants. Eighteen months old drought tolerant and susceptible tea cultivars were each separately exposed to water stress or control conditions of 18 or 34% soil moisture content, respectively, for three months. After the treatment period, leaves were harvested from each treatment for isolation of RNA and cDNA synthesis. The cDNA libraries were sequenced on Roche 454 high-throughput pyrosequencing platform to produce 232,853 reads. After quality control, the reads were assembled into 460 long transcripts (contigs). The annotated contigs showed similarity with proteins in the Arabidopsis thaliana proteome. Heat shock proteins (HSP70), superoxide dismutase (SOD), catalase (cat), peroxidase (PoX), calmodulinelike protein (Cam7) and galactinol synthase (Gols4) droughtrelated genes were shown to be regulated differently in tea plants exposed to water stress. HSP70 and SOD were highly expressed in the drought tolerant cultivar relative to the susceptible cultivar under drought conditions. The genes and pathways identified suggest efficient regulation leading to active adaptation as a basal defense response against water stress deficit by tea. The knowledge generated can be further utilized to better understand molecular mechanisms underlying stress tolerance in tea.

Hepatic Expression of Cu/Zn-Superoxide Dismutase Transcripts in Response to Acute Metal Exposure and Heat Stress in Hemibarbus mylodon (Teleostei: Cypriniformes)

  • Cho, Young-Sun;Bang, In-Chul;Lee, Il-Ro;Nam, Yoon-Kwon
    • Fisheries and Aquatic Sciences
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    • v.12 no.3
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    • pp.179-184
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    • 2009
  • Hemibarbus mylodon (Cypriniformes) is an endemic freshwater fish species in the Korean peninsula, for which urgent conservation efforts are needed. To understand their stress responses in relation to metal toxicity and thermal elevation, we performed a real-time RT-PCR-based expression assay of hepatic copper/zinc-superoxide dismutase (Cu/Zn-SOD), a key antioxidant enzyme, in response to experimental heavy metal exposure or heat treatment. The transcription of hepatic Cu/Zn-SOD was differentially modulated by acute exposure to Cu, cadmium (Cd), or Zn. Exposure to each metal at $5{\mu}M$ for 24 h revealed that Cu stimulated the mRNA expression of Cu/Zn-SOD to a greater extent than the other two heavy metals. The elevation in Cu/Zn-SOD transcripts in response to Cu exposure was dose-dependent (0.5 to $5{\mu}M$). Time course analysis of Cu/Zn-SOD expression in response to Cd exposure ($5{\mu}M$) revealed a transient pattern up to day 7. Exposure to thermal stress (an increase from 22 to $30^{\circ}C$ at a rate of $1^{\circ}C/h$ followed by $30^{\circ}C$ for 18 h) did not significantly alter SOD transcription, although heat shock protein 90 kDa (HSP90) transcription was positively correlated with an increase in temperature.

Effects of Hyperbaric Pressure on Cellular Morphology, Proliferation and Protein Expression of Jurkat Cell

  • Oh, Eun-Ha;Oh, Sang-Nam;Im, Ho-Sub;Lee, Joo-Hyun;Kim, Jin-Young;Moon, Joo-Hee;Hong, Eun-Young;Kim, Yang-Hee;Yang, Min-Ho;Lim, Yong-Chul;Park, Sun-Young;Lee, Eun-Il;Sul, Dong-Geun
    • Molecular & Cellular Toxicology
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    • v.1 no.2
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    • pp.116-123
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    • 2005
  • The application of high pressure on cellular morphology, proliferation and protein expression of Jurkat cells (human T lymphocyte cell line) has been extensively investigated. In the present study, we manufactured a novel pressure chamber that modulates 5% $CO_{2}$, temperature and pressure (up to 3 ATA). Jurkat cells was incubated 2 ATA pressure and analyzed cellular morphology and growth using an electron microscopy and MTT assay. The cells showed the morphological changes in the cell surface, which appeared to cause a severe damage in cell membrane. The growth rate of the cells under 2 ATA pressure decreased as cultured time got increased. Furthermore, a long term exposure of high pressure on Jurkat cells may act as one of the important cellular stresses that leads to inducing cell death. Cellular proteomes were separated by 2-dimensional electrophoresis with pH 3-10 ranges of IPG Dry strips. And many proteins showed significant up-and-down expressions with hyperbaric pressure. Out of all, 10 spots were identified significantly using matrix-assisted laser desorption/ionization-time of fight (MALDI-TOF) mass spectrometry. We and found that 9 protein expressions were decreased and one protein, heat shock protein HSP 60, was increased in Jurkat cells under 2 ATA. Identified proteins were related to lipid metabolism and signal transduction.

Hematological Parameters and Stress Responses of Olive Flounder Paralichthys olivaceus by Acute pH Change (pH의 급성 변화에 따른 넙치(Paralichthys olivaceus)의 혈액학적 성상 및 스트레스 반응)

  • Oh, Min Hyeok;Kim, Jun Young;Kim, Seok-Ryel;Kim, Su Kyoung;Kim, Jun-Hwan
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.53 no.5
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    • pp.733-739
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    • 2020
  • Olive flounder Paralichthys olivaceus (total weight 216.4±14.6 g, total length 28.4±1.7 cm) were exposed to different pH levels (3, 4, 5, 6, 7, 8, and 9) for 96 h. At pH 4, hemoglobin decreased significantly, while plasma calcium, glucose, cholesterol, and ALP increased significantly. Exposure to pH 4 also induced stress responses, as evidenced by a significant decrease in heat shock protein 70 (HSP 70) and a significant increase in cortisol. The results of this study indicate that acute exposure to acidic or alkaline pH (pH 3 or 9) induced significant mortality, while exposure to pH 4significantly affected hematological parameters and stress responses in P. olivaceus.

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.

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.

Transduced Tat-α-Synuclein Protects against Oxidative Stress In vitro and In vivo

  • Choi, Hee-Soon;Lee, Sun-Hwa;Kim, So-Young;An, Jae-Jin;Hwang, Seok-Il;Kim, Dae-Won;Yoo, Ki-Yeon;Won, Moo-Ho;Kang, Tae-Cheon;Kwon, Hyung-Joo;Kang, Jung-Hoon;Cho, Sung-Woo;Kwon, Oh-Shin;Choi, Jin-Hi;Park, Jin-Seu;Eum, Won-Sik;Choi, Soo-Young
    • BMB Reports
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    • v.39 no.3
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    • pp.253-262
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    • 2006
  • Parkinson's disease (PD) is a common neurodegenerative disorder and is characterized by the progressive loss of dopaminergic neurons in the substantia nigra. Although many studies showed that the aggregation of $\alpha$-synuclein might be involved in the pathogenesis of PD, its protective properties against oxidative stress remain to be elucidated. In this study, human wild type and mutant $\alpha$-synuclein genes were fused with a gene fragment encoding the nine amino acid trans activator of transcription (Tat) protein transduction domain of HIV-l in a bacterial expression vector to produce a genetic in-frame WT Tat-$\alpha$-synuclein (wild type) and mutant Tat-a-synucleins (mutants; A30P and A53T), respectively, and we investigated the protective effects of wild type and mutant Tat-$\alpha$-synucleins in vitro and in vivo. WT Tat-$\alpha$-synuclein rapidly transduced into an astrocyte cells and protected the cells against paraquat induced cell death. However, mutant Tat-$\alpha$-synucleins did not protect at all. In the mice models exposed to the herbicide paraquat, the WT Tat-$\alpha$-synuclein completely protected against dopaminergic neuronal cell death, whereas mutants failed in protecting against oxidative stress. We found that these protective effects were characterized by increasing the expression level of heat shock protein 70 (HSP70) in the neuronal cells and this expression level was dependent on the concentration of transduced WT Tat-$\alpha$-synuclein. These results suggest that transduced Tat-$\alpha$-synuclein might protect cell death from oxidative stress by increasing the expression level of HSP70 in vitro and in vivo and this may be of potential therapeutic benefit in the pathogenesis of PD.

HSP90 inhibitor, AUY922, debilitates intrinsic and acquired lapatinib-resistant HER2-positive gastric cancer cells

  • Park, Kang-Seo;Hong, Yong Sang;Choi, Junyoung;Yoon, Shinkyo;Kang, Jihoon;Kim, Deokhoon;Lee, Kang-Pa;Im, Hyeon-Su;Lee, Chang Hoon;Seo, Seyoung;Kim, Sang-We;Lee, Dae Ho;Park, Sook Ryun
    • BMB Reports
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    • v.51 no.12
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    • pp.660-665
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    • 2018
  • Human epidermal growth factor receptor 2 (HER2) inhibitors, such as trastuzumab and lapatinib are used to treat HER2-positive breast and gastric cancers. However, as with other targeted therapies, intrinsic or acquired resistance to HER2 inhibitors presents unresolved therapeutic problems for HER2-positive gastric cancer. The present study describes investigations with AUY922, a heat shock protein 90 (HSP90) inhibitor, in primary lapatinib-resistant (ESO26 and OE33) and lapatinib-sensitive gastric cancer cells (OE19, N87, and SNU-216) harboring HER2 amplification/over-expression. In order to investigate whether AUY922 could overcome intrinsic and acquired resistance to HER2 inhibitors in HER2-positive gastric cancer, we generated lapatinib-resistant gastric cancer cell lines (OE19/LR and N87/LR) by continuous exposure to lapatinib in vitro. We found that activation of HER2 and protein kinase B (AKT) were key factors in inducing intrinsic and acquired lapatinib-resistant gastric cancer cell lines, and that AUY922 effectively suppressed activation of both HER2 and AKT in acquired lapatinib-resistant gastric cancer cell lines. In conclusion, AUY922 showed a synergistic anti-cancer effect with lapatinib and sensitized gastric cancer cells with intrinsic resistance to lapatinib. Dual inhibition of the HSP90 and HER2 signaling pathways could represent a potent therapeutic strategy to treat HER2-positive gastric cancer with intrinsic and acquired resistance to lapatinib.

Hsp90 Inhibitor Induces Cell Cycle Arrest and Apoptosis of Early Embryos and Primary Cells in Pigs

  • Son, Myeong-Ju;Park, Jin-Mo;Min, Sung-Hun;Hong, Joo-Hee;Park, Hum-Dai;Koo, Deog-Bon
    • Reproductive and Developmental Biology
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    • v.35 no.1
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    • pp.33-45
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    • 2011
  • Heat shock protein 90 (Hsp90) is ATPase-directed molecular chaperon and affects survival of cancer cell. Inhibitory effect of Hsp90 by inducing cell cycle arrest and apoptosis in the cancer cell was reported. However, its role during oocyte maturation and early embryo development is very insufficient. In this study, we traced the effects of Hsp90 inhibitor, 17-allylamino-17-demethoxygeldanamycin (17-AAG), on meiotic maturation and early embryonic development in pigs. We also investigated several indicators of developmental potential, including structural integrity, gene expression (Hsp90-, cell cycle-, and apoptosis-related genes), and apoptosis, which are affected by 17-AAG. Then, we examined the roles of Hsp90 inhibitor on viability of primary cells in pigs. Porcine oocytes were cultured in the NCSU-23 medium with or without 17-AAG for 44 h. The proportion of GV arrested oocytes was significantly different between the 17-AAG treated and untreated group (78.2 vs 34.8%, p<0.05). After completion of meiotic maturation, the proportion of MII oocytes was lower in the 17-AAG treated group than in the control group (27.9 vs 71.0%, p<0.05). After IVF, the percentage of penetrated oocytes was significantly lower in the 17-AAG treated group (25.2%), resulting in lower normal pronucleus formation (2PN of 14.6%). Therefore, the inhibition of meiotic progression by Hsp90 inhibitor played a critical role in fertilization status. Porcine embryo were cultured in the PZM-3 medium with or without 17-AAG for 6 days. In result, significant differences in developmental potential were detected between the embryos that were cultured with or without 17-AAG. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) showed that the number of containing fragmented DNA at the blastocyst stage increased in the 17-AAG treated group compared with control (7.5 vs 4.4, respectively). Blastocysts that developed in the 17-AAG treated group had low structural integrity and high apoptotic nuclei than those of the untreated control, resulting in decrease the embryonic qualities of preimplantation porcine blastocysts. The mRNA expressions of cell cycle-related genes were down-regulated in the 17-AAG treated group compared with control. Also, the expression of the pro-apoptotic gene Bax increased in 17-AAG treated group, whereas expression of the anti-apoptotic gene Bel-XL decreased. However, the expression of ER stress-related genes did not changed by 17-AAG. Cultured pESF cells were treated with or without 17-AAG and used for MIT assay. The results showed that viability of pESF cells were decreased by treatment of 17-AAG ($2{\mu}M$) for 24 hr. These results indicated that 17-AAG decreased cell proliferation and increased cell death. Expression patterns Hsp90 complex genes (Hsp70 and p23), cell cycle-related genes (cdc2 and cdc25c) and apoptosis-related genes (Bax and Bcl-XL) were significantly changed by using RT-PCR analysis. The spliced form of pXbp-1 product (pXbp-1s) was detected in the tunicamycin (TM) treated cells, but it is not detected in 17-AAG treated cells. In conclusion, Hsp90 appears to play a direct role in porcine early embryo developmental competence including structural integrity of blastocysts. Also, these results indicate that Hsp90 is closely associated with cell cycle- and apoptosis-related genes expression in developing porcine embryos.

KISS Korea Computer Congress 2007 (이동 객체의 패턴 탐사를 위한 시공간 데이터 일반화 기법)

  • Ko, Hyun;Kim, Kwang-Jong;Lee, Yon-Sik
    • Proceedings of the Korean Information Science Society Conference
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    • 2007.06c
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    • pp.153-158
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    • 2007
  • 사용자들의 특성에 맞게 개인화되고 세분화된 위치 기반 서비스를 제공하기 위해서는 방대한 이동 객체의 위치 이력 데이터 집합으로부터 유용한 패턴을 추출하여 의미 있는 지식을 탐사하기 위한 시공간 패턴 탐사가 필요하다. 현재까지 다양한 패턴 탐사 기법들이 제안되었으나 이동 패턴들 중 단순히 시공간 제약이 없는 빈발 패턴만을 추출하기 때문에 한정된 시간 범위와 제한적인 영역 범위 내에서의 빈발 패턴을 탐사하는 문제에는 적용하기 어렵다. 또한 패턴 탐사 수행 시 데이터베이스를 반복 스캔하여 탐사 수행시간이 많이 소요되는 문제를 포함하거나 메모리상에 탐사 대상인 후보 패턴 트리를 생성하는 방법을 통해 탐사 시간을 줄일 수는 있으나 이동 객체 수나 최소지지도 등에 따라 트리를 구성하고 유지하는데 드는 비용이 커질 수 있다. 따라서 이러한 문제를 해결하기 위한 효율적인 패턴 탐사 기법의 개발이 요구됨으로써 선행 작업으로 본 논문에서는 상세 수준의 객체 이력 데이터들의 시간 및 공간 속성을 의미 있는 시간영역과 공간영역 정보로 변환하는 시공간 데이터 일반화 방법을 제안한다. 제안된 방법은 공간 개념 계층에 대한 영역 정보들을 영역 Grid 해쉬 테이블(AGHT:Area Grid Hash Table)로 생성하여 공간 인덱스트리인 R*-Tree의 검색 방법을 이용해 이동 객체의 위치 속성을 2차원 공간영역으로 일반화하고, 시간 개념 계층을 생성하여 이동 객체의 시간적인 속성을 시간 영역으로 일반화함으로써 일반화된 데이터 집합을 형성하여 효율적인 이동 객체의 시간 패턴 마이닝을 유도할 수 있다.의 성능을 기대할 수 있을 것이다.onium sulfate첨가배지(添加培地)에서 가장 저조(低調)하였다. vitamin중(中)에서는 niacin과 thiamine첨가배지(添加培地)에서 근소(僅少)한 증가(增加)를 나타내었다.소시켜 항이뇨 및 Na 배설 감소를 초래하는 작용과, 둘째는 신경 경로를 통하지 않고, 아마도 humoral factor를 통하여 신세뇨관에서 Na 재흡수를 억제하는 작용이 복합적으로 나타내는 것을 알 수 있었다.으로 초래되는 복합적인 기전으로 추정되었다., 소형과와 기형과는 S-3에서 많이 나왔다. 이상 연구결과에서 입도분포가 1.2-5mm인 것이 바람직한 것으로 나타났다.omopolysaccharides로 확인되었다. EPS 생성량이 가장 좋은 Leu. kimchii GJ2의 평균 분자량은 360,606 Da이었으며, 나머지 두 균주에 대해서는 생성 EPS 형태와 점도의 차이로 미루어 보아 생성 EPS의 분자구조와 분자량이 서로 다른 것으로 판단하였다.TEX>개로 통계학적으로 유의한 차이가 없었다. Heat shock protein-70 (HSP70)과 neuronal nitric oxide synthase (nNOS)에 대한 면역조직화학검사에서 실험군 Cs2군의 신경세포가 대조군 12군에 비해 HSP70과 nNOS의 과발현을 보였으며, 이는 통계학적으로 유의한 차이를 보였다(p<0.05). nNOS와 HSP70의 발현은 강한 연관성을 보였고(상관계수 0.91, p=0.000), nNOS를 발현하는 세포가 동시에 HSP70도 발현함을 확인할 수 있었다. 결론: 우리는

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