• Journal of Microbiology and Biotechnology
• /
• 제10권2호
• /
• pp.137-142
• /
• 2000

Environmental stres is known to induce heat shock proteins (HSPs) in eukaryotic cells. However, the induction of HSPs in host cells by microbial infection has not yet been well explained. Staphylococcus aureus (S. aureus) is one of the major pathogens in the pathogenesis of endovascular diseases such as infective endocarditis. In this study, the synthesis of stress-inducible 70 kDa HSP was investigated in the endothelial cells (ECs) after 3 h to 20 h of incubation with S. aureus. The dffect of S. aureus infection on the expression of HSP70 in cultured ECs was analyzed using laser scanning confocal microscopy (LSCM). The increase of HSP70 expression in ECs infected by S. aureus ($10^4{\;}cfu/ml$) for 20 h was 1.1-fold higher than that in heat shock treated ECs and 2.2-fold higher than that in untreated cells. Heat shock is known to induce intranucleus HSP70 expression in mammalian cells, whereas the S. aureus infection induced perinuclear expression in ECs as observed by LSCM. Consequently, the expression of HSP70 in ECs plays an important role in the pathogenesis of endovascular infection.

• 미생물학회지
• /
• 제34권1_2호
• /
• pp.43-50
• /
• 1998

여러가지 환경 stress에서 유도되는 폐렴구균 열충격단백질 GroEL의 특성에 대하여 검토하였다. 비병원성인 폐렴구균(Streptococcusa pneumoniae)을 사용하여 stress 조건을 설정하고 이 설정된 조건에서 stress에 의해 유도되는 단백질을 $^{35}S$]-methionine으로 표지하여 autoradiography를 실시하였다. 열충격을 가했을 때 유도되는 단백질(65, 73, 84-kDa 등) 중 65 kDa의 단백질(hsp65)을 DEAE-Sepharose ion exchange 및 ATP-agarose affinity chromatography를 이용하여 분리 정제하였으며 hsp65에 대해 생성된 항체를 이용하여 immunoblot을 실시하였을 때 약 60 kDa의 대장균 단백질과 반응하였으며 정제된 폐렴구균 hsp65가 대장균의 anti-GroEL monoclonal 항체와 반응함으로써 폐렴구균 hsp65가 대장균GroEL과 유사한 단백질임을 확인하였다.

• Bulletin of the Korean Chemical Society
• /
• 제34권12호
• /
• pp.3782-3786
• /
• 2013

Non-small cell lung cancer (NSCLC) is the most common type of lung cancer and that accounts for 85% of lung cancer patients. Although several EGFR-targeted drugs have been developed in the treatment of NSCLC, the clinical efficacy of EGFR-targeted drugs in NSCLC is limited by the occurrence of drug resistance. In this regard, Hsp90 represents great promise as a therapeutic target of cancer due to its potential to simultaneously disable multiple signaling pathways. In this study, we discovered that a natural product, flavokawain B disrupted Hsp90 chaperoning function and impaired the growth of gefitinib-resistant non-small cell lung cancer (H1975). The result suggested that flavokawain B could serve as a potential lead compound to overcome the drug resistance in cancer chemotherapy.

• 한국동물학회지
• /
• 제30권2호
• /
• pp.177-192
• /
• 1987

온열처리가 근세포의 분화에 어떤 영향을 미치는 지를 알아보기 위하여 배양한 근세포에 여러가지 온열처리를 한 다음, 단백질 합성, 세포증식, 융합지수, creatine kinase(CK)의 활성 및 cholesterol 함량의 변화를 조사하였다. 배양한지 24시간지나 45$^{\circ}C$에서 1 hr의 온열처리를 하면 근세포의 융합과 CK 활성이 지연되었으며, 55시간지나 같은 온열처리를 하면 세포막내(세포내 양의 95% 이상)의 chloesterol 양이 일시적으로 증가함과 아울러 세포융합이 지연되었다. 그러나 세포증식은 대조군과 뚜렷한 차이가 없었다. 이상과 같은 실험 결과로부터 온열처리가 분화에 미치는 영향은 온열처리를 받게되는 근원세포의 분화정도에 따라 차이가 있으며 온열처리에 따른 chloesterol 양의 일시적인 증가가 근세포 융합에 영향을 미칠 수 있다는 가능성이 제시되었다. 한편, 근세포는 온열처리를 받으면 평소의 단백질 합성 수준이 떨어짐과 더불어 heat shock protein(HSP)합성이 증대 내지는 유도 되었으며 HSP 합성의 유도는 actinomycin D의 처리로 억제되었다. 또한 온열처리로 근세포는 열내성을 얻어 세포융하과 CK 활성은 동일한 온열처리를 4시간 간격으로 두번 주어도 한번 주었을 경우와 차이가 없었으며 두번째 온열처리에 의해서는 새로운 HSP 합성이 유도되지도 않았다.

• 한국식품위생안전성학회지
• /
• 제13권2호
• /
• pp.171-187
• /
• 1998

Garlic occupies a special position among the many foods of vegetable origin because it is the sole food for Koreans during the their lives. And vitamin A has been ingested by forms of food or additives. Cadmium has been described as one of the most dangerous trace elements in the food and environment of man and livestocks. Since the de novo synthesis of stress proteins can be detected early after exposure to some agents, analysis of cadmium-induced changes in gene expression , ie. alterations in patterns of protein synthesis, may be useful to develop as biomarkers of exposure and damage for food hygiene. He acute and chronic combine effects of cadmium (Cd, CdCl2 20mg/kg), garlic oil(Dds: diallyl disulfide 50mg/kg, 3 times a week) and vitamin A(Ra: retinol acetate 50,000 IU/kg, 3 times a week) on Wistar male rats were evaluated concerning cadmium contents, tissues enzyme activity, HSP expression histopathological and electron microscopical examinations. The results of the study are as follows ; 1. Less cadmium was absorbed through the digestive tracts, but the ratio of contents in tissue were not changed by the simultaneous adminstration of diallyl disufide or retinol acetate. 2. ALT(alanine aminotransferase) , AST(aspartate aminotransferase), glucose, BUN (blood urea nitrogen), creatinine, the key indices of the clinical changes in hepatic and renal function were significantly hanged by the cadmium treatment after 1 week in liver, after 4 weeks in kidney. 3. Histopathological changes in cadmium treated rats were appeared at 8 weeks age treatment in kidneys. Homogenous eosinophilic material was accumulated in cortical and collecting tubular lumens at 16 weeks. Degenerated or necrotized tubular cells were observed in cortex and medulla. Degenerated seminiferous tubules and homogeneous eosinophilic material was seen in interstitial tissue of rat treated with cadmium for 16 weeks. Calcium deposits were seen in degenerated seminiferous tubules and the tubules showed severe calcification of rat treated with cadmium for 16 weeks. Electron microscope changes in kidney were observed in rats treated with CdCl2 20 mg/kg. Proximal convoluted tubule cells showed selling of cytoplasm and narrow lumen. Capillary endothelial cells showed cytoplasmic vacuoles and swelling. Degenerated epithelial cells were accumulated in tubular lumen of kidney. 4. Enhanced synthesis of 70 KDa relateve molecular mass proteins were detected in 2 hours after cadmium, exposure, with maximum activity occurring at 8~48 hours. Induction of HSP 70 was evident at proximal tubules and glomeruli in kidney. Testicular cells produced enough HSP to be detected normally. From the above results, it could be concluded that HSP70 induction by the cadmium treatment was a rapid reaction to indicated the exposure of xenobiotics, and retinol acetate reduced the cadmium induced nephrotoxicity.

• Journal of Animal Science and Technology
• /
• 제64권1호
• /
• pp.110-122
• /
• 2022

The reduction of milk yield caused by heat stress in summer is the main condition restricting the economic benefits of dairy farms. To examine the impact of hyperthermia on bovine mammary epithelial (MAC-T) cells, we incubated the MAC-T cells at thermal-neutral (37℃, CON group) and hyperthermic (42℃, HS group) temperatures for 6 h. Subsequently, the cell viability and apoptotic rate of MAC-T cells, apoptosis-related genes expression, casein and amino acid transporter genes, and the expression of the apoptosis-related proteins were examined. Compared with the CON group, hyperthermia significantly decreased the cell viability (p < 0.05) and elevated the apoptotic rate (p < 0.05) of MAC-T cells. Moreover, the expression of heat shock protein (HSP)70, HSP90B1, Bcl-2-associated X protein (BAX), Caspase-9, and Caspase-3 genes was upregulated (p < 0.05). The expression of HSP70 and BAX (pro-apoptotic) proteins was upregulated (p < 0.05) while that of B-cell lymphoma (BCL)2 (antiapoptotic) protein was downregulated (p < 0.05) by hyperthermia. Decreased mRNA expression of mechanistic target of rapamycin (mTOR) signaling pathway-related genes, amino acid transporter genes (SLC7A5, SLC38A3, SLC38A2, and SLC38A9), and casein genes (CSNS1, CSN2, and CSN3) was found in the heat stress (HS) group (p < 0.05) in contrast with the CON group. These findings illustrated that hyperthermia promoted cell apoptosis and reduced the transport of amino acids into cells, which inhibited the milk proteins synthesis in MAC-T cells.

• 대한본초학회지
• /
• 제20권4호
• /
• pp.1-10
• /
• 2005

Objectives : We investigated the anti-aging effect on skin with the extract of the needles of red pine, Pinus densiflora. Methods : We measured various effects related to skin such as the anti-oxidant effect, the protection against ultraviolet (UV) irradiation, the inhibition of reactive oxygen species (ROS) generation, the induction of heat shock protein 70 (HSP70), the reduction of matrix metalloproteinase-2 (MMP-2) synthesis and senescent cell. Results : The results were as follows : The extract of the needles of red pine (RP) had the potent anti-oxidant effect and the ROS scavenging effect. Also RP preserved the systemic anti-oxidant enzyme system (superoxide dismutase and catalase) from UVB irradiation. RP protected the cell membrane from the damages induced by UVB irradiation. RP induced HSP70, a mediator of resistance to UVB irradiation. RP reduced the synthesis of MMP-2 induced by UVB irradiation. And RP inhibited the amount of senescent-associated (SA) ${\beta}-galactosidase$ staining, as a marker of replicative senescence. Conclusions : The results of our study indicate that the extract of the needles of red pine, Pinus densiflora, has anti-aging effects on skin.

• Journal of Microbiology
• /
• 제34권1호
• /
• pp.37-37
• /
• 1996

The internal regions of nuclear small subunit rRNA from 6 plaeurotus species and 5 Pleurotus ostreatus strains were amplified by PCR and sequenced. The DNA sequences of 8 Pleurotus strains (P. ostreatus NFFA2, NFFA4501, NFFA4001, KFFA4001, KFCC11635, P florida, P. florida, P. sajor-cuju, P. pulmonarius, and P. spodoleucus) were idential, but P. cornucopiae differed from them in two bases out of 605 bases. However, p[hylogenetic analysis of the sequences by DNA-distance matrix and UPGMA methods showed that P. ostreatus NFFA2m1 and NFFA2m2, known as mutants of P. ostreatus NFFA2, belonged to anther group of Basidiomycotina, which is close to the genus Auricularia. The difference of the SSU rDNA sequences of P. cornucopiae from other Pleurotus species tested corresponds to the difference of mitochondrial plasmid type present in Pleurotus species as observed by Kim et al. (1993, Korean J. Microbiol. 31, 141-147).ishement of silencing at the HMR/hsp82 locus can occur in G1-arrested cells. Cell cycle arrest at G1 phase was achieved by treatment of early log a cell cultures with .alpha.-factor mating pheromone, which induces G1 arrest. The result suggests that passage through S phase (and therefore DNA replication) is nor required for re-establishing silencer-mediated repression at the HMNRa/HSP82 locus. Finally, to test whether de nono protein synthesis is required for re-establishment of silencer-mediated repression, cells were pretreated with cycloheximide (500 /.mu.g/ml) 120 min. It was apparent that inhibiting protein synthesis delays, but does not prevent, re-establishment of silencer-mediated repression. Altogether, these results indicate that re-establishment of silencer-mediated repression is not dependent on the DNA replication and has no requirement for protein synthesis.

• 한국수정란이식학회:학술대회논문집
• /
• 한국수정란이식학회 2002년도 국제심포지엄
• /
• pp.78-78
• /
• 2002

The role of heat shock proteins in shielding organism from environmental stress is illustrated by the large-scale synthesis of these protein by the organism studied to date. However, recent evidence also suggests an important role for heat shock protein in fertilization and early development of mammalian embryos. Effects of elevated in vitro temperature on in vitro produced bovine embryos were analysed in order to determine its impact on the expression of heat shock protein 70 (HSP70) by control and frozen-thawed after in vitro fertilization (IVF) or nuclear transfer (NT). The objective of this study was to assess the developmental potential in vitro produced embryos with using of the various containers and examined expression and localization of heat shock protein 70 after it's frozen -thawed. For the vitrification, in vitro produced embryos at 2 cell, 8 cell and blastocysts stage after IVF and NT were exposed the ethylene glycol 5.5 M freezing solution (EG 5.5) for 30 sec, loaded on each containers such EM grid, straw and cryo-loop and then immediately plunged into liquid nitrogen. Thawed embryos were serially diluted in sucrose solution, each for 1 min, and cultured in CRI-aa medium. Survival rates of the vitrification production were assessed by re-expanded, hatched blastocysts. There were no differences in the survival rates of IVF using EM grid, cryo-loop. However, survival rates by straw were relatively lower than other containers. Only, nuclear transferred embryos survived by using cryo-loop. After IVF or NT, in vitro matured bovine embryos 2 cell, 8 cell and blastocysts subjected to control and thawed conditions were analysed by semiquantitive reverse transcription polymerase chain reaction methods for hsp 70 mRNA expression. Results revealed the expression of hsp 70 mRNA were higher thawed embryos than control embryos. Immunocytochemistry used to localization the hsp70 protein in embryos. Two, 8-cell embryos derived under control condition was evenly distributed in the cytoplasm but appeared as aggregates in some embryos exposed frozen-thawed. However, under control condition, blastocysts displayed aggregate signal while Hsp70 in frozen-thawed blastocysts appeared to be more uniform in distribution.

• The Korean Journal of Physiology and Pharmacology
• /
• 제1권6호
• /
• pp.775-781
• /
• 1997

Although reoxygenation is the best way to salvage hypoxic tissues, reduced oxygen species (ROS) generated during reoxygenation are blown to cause further tissue injuries and the induction of heat shock proteins (HSPs). The present study was undertaken to determine any causal relationship between the severity of hypoxia and the opposite outcomes, either beneficial or detrimental, of the subsequent reoxygenation by measuring the HSP72. To this aim, one group (6 male cats, $2.5{\sim}3.5\;kg$) was subjected to a 5-min episode of hypoventilation (H, ventilation rate: 5/min) for the induction of slight hypoxia and the other group (6 male cats, $2.4{\sim}3.7\;kg$) was subjected to a 5-min episode of apnea (A) for severe hypoxia. Each 3 animals from both groups received a 10-min episode of ventilation with $(95%\;O_2\;(0)$, whereas the remainder did not. After these procedures, all animals were allowed to be ventilated within physiological range for 1, 4, or 8 hours (1H, 1HO, 4H, 4HO, 8H, 8HO, 1A, 1AO, 4A, 4AO, 8A and 8AO groups). Control animals did not receive any manipulation. The arterial blood $pCO_2$ was significantly higher just after apnea than hypoventilation, while $pCO_2$ and pH were significantly lower just after apnea than hypoventilation. Western blot analysis revealed that the magnitude of HSP72 synthesis is larger in 1H, 4H and 8H groups than in 1HO, 4H and 8HO groups, respectively. In contrast, 1AO, 4AO and 8AO groups more induced HSP72 than 1A, 4A and 8A groups, respectively. These results suggest that the reoxygenation is beneficial after slight hypoxia but detrimental after severe hypoxia.