• Maxillofacial Plastic and Reconstructive Surgery
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• v.20 no.4
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• pp.362-372
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• 1998

Heat shock protein (HSP) expression is unregulated in tumor cells and, HSP expression is likely marker of the malignant potential of oral epithelial lesion. Furthermore, the 70kDa HSP is implicated in the degree of tumor differentiation, the rate of tumor proliferation and the magnitude of the anti-tumor immune response. Accordingly, the distribution and intensity of HSP 70 and HSP 47 expression was assessed in the DMBA induced oral carcinogenesis in hamster. Golden Syrian hamsters which were 3 months-age and 90-120g were collected. 9,10-dimethyl-1,2-benzanthracene (DMBA) in a 0.5% solution in mineral oil was painted on the buccal pouch mucosa 3 times per week in the study group. In each control and experimental groups of 6, 8, 10, 12, 14, 16, 18, 20 weeks, specimen were sectioned for immunohistochemical study with anti-HSP47 and anti-HSP70 antibody. The following results were obtained. 1. HSP47 positive cells were rare or negative of normal oral mucosa, increased mildly in basal and suprabasal basal layer, and spinous cell layer after experimental 6 weeks (dysplastic or CIS stage). In CIS stage, HSP47 expression is prominent in dysplastic free or normal adjacent epithelium. 2. HSP 47 positive cells in connective tissue were mainly inflammatory cells, which is gradually increased from control to precancerous and cancer stage. But HSP47 positive cells after 14 weeks were decreased, especially normal and cancer adjacent epithelium. 3. The positive staining cells of HSP70 in control, dysplastic, and CIS stage were not seen. But they were mild findings in basal layer and moderate findings in spinous layer after experimental 14 weeks (cancer stage). 4. HSP70 positive cells were increased in precancerous and cancer stage than control group in connective tissue. After experimental 16 weeks, we could not find the HSP expression in cancer cells according to cancer differentiation or cancer stage. It is concluded that HSP70 or HSP47 expression is not a definitive marker of oral malignancy or malignant potential. However, with further development, HSP immunoreactivity may be valuable as an adjunct to conventional histology for assessing the malignant potential of oral mucosal lesions.

• The Journal of Korean Physical Therapy
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• v.20 no.1
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• pp.57-65
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• 2008

Purpose: Heat shock proteins (HSPs) are a group of proteins that are activated when cells are exposed to a variety of environmental stresses, such as infection, inflammation, exposure to toxins, starvation, hypoxia, brain injury, or water deprivation. The activation of HSPs by environmental stress plays a key role in signal transduction, including cytoprotection, molecular chaperone, anti-apoptotic effect, and anti-aging effects. However, the precise mechanism for the action of small HSPs, such as HSP27 and mitogen-activated protein kinases (MAPKs: extracellular-regulated protein kinase 1/2 (ERK1/2), p38MAPK, stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK), is not completely understood, particularly in application of cell stimulators including platelet-derived growth factor (PDGF), angiotensin II (AngII), tumor necrosis factor $\alpha$ (TNF$\alpha$), and $H_2O_2$. This study examined the relationship between stimulators-induced enzymatic activity of HSP27 and MAPKs from rat smooth and skeletal muscles. Methods: 2-dimensional electrophoresis (2DE) and matrix assisted laser desorption ionizationtime-of-flight/time-of-flight (MALDI-TOF/TOF) analysis were used to identify HSP27 from the intact vascular smooth and skeletal muscles. Three isoforms of HSP27 were detected on silver-stained gels of the whole protein extracts from the rat aortic smooth and skeletal muscle strips. Results: The expression of PDGF, AngII, TNF$\alpha$, and $H_2O_2$-induced activation of HSP27, p38MAPK, ERK1/2, and SAPK/JNK was higher in the smooth muscle cells than the control. SB203580 (30${\mu}$M), a p38MAPK inhibitor, increased the level of HSP27 phosphorylation induced by stimulators in smooth muscle cells. Furthermore, the age-related and starvation-induced activation of HSP27 was higher in skeletal muscle cells (L6 myoblast cell lines) and muscle strips than the control. Conclusion: These results suggest, in part, that the activity of HSP27 and MAPKs affect stressors, such as PDGF, AngII, TNF$\alpha$, $H_2O_2$, and starvation in rat smooth and skeletal muscles. However, more systemic research will be needed into physical therapy, including thermotherapy, electrotherapy, radiotherapy and others.

• Korean Journal of Ecology and Environment
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• v.50 no.3
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• pp.305-313
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• 2017

We investigate that the impact of freshwater organism exposed to the salinity environment by the frequent rainfall following climate change. To evaluate the stress response following salinity exposure, we assessed the survival rate, molting success rate, the developmental period and mouthpart deformities in Chironomus riparius. In addition, we measured the molecular responses of biomarker gene, gene expression of heat shock protein 70 (HSP70) in C. riparius exposed to salinity after 96 hour. The C. riparius survival rates were showed on time dependent manner and not observed survival organisms above 15 psu at day 4. The pupation and emergence of C. riparius were not seen above 15 psu, and the molting success rate was less than 20% at 10 psu. The developmental retardation of C. riparius was well observed in the pupation and emergence period and was delayed by 4 days at 10 psu compared to the control and 5 psu. The mouthpart deformities after salinity exposure at 96 or 72 hour were observed at 10 psu and 15 psu. The expression of C. riparius HSP70 level was significantly increased exposure to 5 psu and 10 psu. Thus, salinity has been caused to be various ecotoxicological and molecular stress responses on freshwater organisms similar to harmful substances such as EDCs and so on.

• Korean Journal of Plant Tissue Culture
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• v.24 no.5
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• pp.295-303
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• 1997

To investigate the response on feeder cell cultures, protoplasts isolated from cell suspensions initiated from mature seed scutellum-derived callus of the Japonica rice variety Taipei 309, were cultured on filter membranes under various conditions. The effects of various factors, such as gelling agents, feeder cell and protoplast densities, species of feeder cells and heat shock treatment, have been investigated to improve protoplast plating efficiencies on filter membranes. Maximum protoplast plating efficiencies were obtained when protoplasts were cultured on KPR medium semi-solidified with Sea Plaque agarose at a density of $5\;\times\;10^{5}\;ml^{-1}$ protoplasts in the presence of Lolium multiflorum as feeder cells (0.5 ml pcv per 10 ml of protoplast culture medium). Pre-culture heat shock treatments for 1 min. and 5 min. to the protoplasts did not give any appreciable increase on the plating efficiency of protoplasts in the presence of feeder cells. Maltose-supplemented medium was superior for plant regeneration from protoplast-derived colonies compared with medium containing only sucrose. The plants were transferred to the glasshouse, flowered and were fertile.

• Korean Journal of Veterinary Research
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• v.45 no.1
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• pp.45-53
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• 2005

GroE that is a heat shock protein composed of GroEL and GroES is known as an immunodominant target of both the humoral and cellular immune responses in bovine brucellosis. This study was carried out to characterize groE gene encoding heat shock proteins of B. abortus isolated in Korea and to evaluate the immunogenicity of the GroE protein expressed in E. coli system. In PCR the specific signals with the size of 2,077 bp were detected in five strains isolated from the mammary lymphnodes of the dairy cattle that were serologically positive and the reference strains. In comparison of the sequences of nucleotides and amino acids among the strains, GroES showed 100% identity in both sequences. GroEL was evaluated 99.0~99.9% in nucleotides and 98.0~100% homology in amino acids. The groE gene including groES and groEL was inserted into pET29a vector and constructed pET29a-GroE recombinant plasmids. The inserted groE was confirmed by digestion with Nco1 and EcoR1 endonucleases and nucleotide sequencing. E. coli BL (DE3) was transformed with pET29a-GroE, named as E. coli BL (DE3)/pET29a-GroE. In SDS-PAGE, it was evident that the recombinant plasmid effectively expressed the polypeptides for GroES (10 kDa) and GroEL (60 kDa) in 0.5, 1 and 2 hours after IPTG induction. The immuno-reactivity of the expressed proteins were proved in mouse inoculation and Western blot analysis.

• International Journal of Industrial Entomology and Biomaterials
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• v.28 no.2
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• pp.71-84
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• 2014

The full-length heat shock protein 88 (HSP88) complementary DNA (cDNA) of Paecilomyces tenuipes Jocheon-1 was obtained by screening the Paecilomyces tenuipes (P. tenuipes) Jocheon-1 Uni-Zap cDNA library and performing 5' RACE polymerase chain reaction (PCR). The P. tenuipes Jocheon-1 HSP88 cDNA contained an open reading frame (ORF) of 2,139-basepair encoding 713 amino acid residues. The deduced amino acid sequence of the P. tenuipe s Jocheon-1 HSP88 cDNA showed 77% identity to Nectria haematococca HSP88 and 45-76% identity to other fungal homologous HSP88s. Phylogenetic analysis and BLAST program analysis confirmed that the deduced amino acid sequences of the P. tenuipes Jocheon-1 HSP88 gene belonged to the ascomycetes group within the fungal clade. The P. tenuipes Jocheon-1 HSP88 also contained the conserved ATPase domain at the N-terminal region. The cDNA encoding P. tenuipes Jocheon-1 HSP88 was expressed as an 88 kilodalton (kDa) polypeptide in baculovirus-infected insect Sf9 cells. Under higher temperature conditions for the growth of the entomopathogenic fungus, mRNA expression of P. tenuipes Jocheon-1 HSP88 was quantified by real time PCR (qPCR). The results showed that heat shock stress induced a higher level of mRNA expression compared to normal growth conditions.

• Journal of Trauma and Injury
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• v.21 no.1
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• pp.59-65
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• 2008

Purpose: The aim of this study was to evaluate the effect of overexpression of heat shock protein 70 (HSP70) on the expression of inducible nitric oxide synthase and on the concentration of nitric oxide and to determine the mechanism for the relationship between HSP70 and inducible nitric oxide synthase (iNOS) in sepsis. Methods: Experiments were performed on male Sprague-Dawley rats, and sepsis was induced by using cecal ligation and puncture (CLP). Glutamine (GLN) or saline was administered 1 h after initiation of sepsis. We acquired serum and lung tissues from the rats 12 h or 24 h after initiation of sepsis. We analyzed the concentration of nitric oxide, the expression of HSP70 in the lung, and the gene expression of iNOS in the lung. Results: In CLP+GLN, glutamine given after initiation of sepsis enhanced the expression of HSP70 in the lung at 12 h (CLP+GLN vs. CLP:: $47.19{\pm}10.04$ vs. $33.22{\pm}8.28$, p = 0.025) and 24 h (CLP+GLN vs. CLP: $47.06{\pm}10.60$ vs. $31.90{\pm}4.83$, p = 0.004). In CLP+GLN, glutamine attenuated the expression of iNOS mRNA in the lung at 12 h (CLP+GLN vs. CLP: $4167.17{\pm}951.59$ vs. $5513.73{\pm}1051.60$, p = 0.025) and 24 h (CLP+GLN vs. CLP: $9,437.65{\pm}2,521.07$ vs. $18,740.27{\pm}8,241.20$, p = 0.016) and reduced the concentration of nitric oxide in serum at 12 h (CLP+GLN vs. CLP: $0.86{\pm}0.48$ vs. $3.82{\pm}2.53{\mu}mol/L$, p = 0.016) and 24 h (CLP+GLN vs. CLP: $0.39{\pm}0.25$ vs. $1.85{\pm}1.70{\mu}mol/L$, p = 0.025). Conclusion: The overexpression of HSP70 induced by the administration of glutamine in sepsis attenuated the gene expression of iNOS and reduced the concentration of nitric oxide.

• Journal of Life Science
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• v.26 no.7
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• pp.826-834
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• 2016

The present investigation was undertaken to examine the effectiveness of the combination treatment of an Hsp90 inhibitor and a SIRT1 inhibitor on suppressing the growth of chemo-resistant human cancer cells. We showed that inhibition of SIRT1 effectively potentiated the cytotoxicity of 17-allylamino-17-demethoxygeldanamycin (17-AAG) and reversed Hsp90 inhibitor resistance in multidrug-resistant (MDR) human ovarian HeyA8-MDR cells. Amurensin G, a potent natural SIRT1 inhibitor, enhanced Hsp90 inhibitor-mediated abrogation of the Hsp90 chaperone function and accelerated degradation of mutated p53 (mut p53), an Hsp90 client protein, by up-regulation of ubiquitin ligase CHIP. Knock-down of CHIP significantly attenuated amurensin G-induced mut p53 degradation. Down-regulation of mut p53 reduced the expression of heat shock factor1 (HSF1)/heat shock proteins (Hsps), a major cause of Hsp90 inhibitor resistance, which led to sensitization of the MDR cells to the Hsp90 inhibitor by the SIRT1 inhibitor. Amurensin G potentiated cytotoxicity of the Hsp90 inhibitor in HeyA8-MDR cells through suppression of 17-AAG-induced Hsp70 and Hsp27 induction via down-regulation of mut p53/HSF1, and it caused activation of PARP and inhibition of Bcl-2. Our data suggests that SIRT1 inhibitors could be used to sensitize MDR cells to Hsp90 inhibitors, possibly through suppression of the mut p53/HSF1-dependent pathway, and a novel mut p53-directed action of SIRT1 inhibition could effectively prevent mut p53 accumulation in MDR cells.

• Journal of Marine Life Science
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• v.3 no.2
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• pp.51-58
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• 2018

Variations in water temperature are known to affect almost every part of fish physiology. The rise in water temperature due to climate change can physically damage fish. This study was conducted to evaluate the health status of the Atlantic salmon (Salmo salar) at high water temperature (20℃) than the optimum water temperature (15℃). Liver tissue exerts important metabolic functions in thermal adaptation. Therefore, liver tissue was used in this study. The evaluation method is to develop the biomarker gene using NGS RNAseq analysis and to examine the expression pattern using RT-qPCR analysis. The NGS RNAseq analysis revealed 1,366 differentially expressed genes, among which 880 genes were increase expressed and 486 genes were decrease expressed. The biomarker genes are such as heat shock protein 90 alpha (Hsp90α), heat shock protein 90 beta (Hsp90β) and cytochrome P450 1A (CYP1A). The selected genes are sensitive to changes in water temperature through NGS RNAseq analysis. Expression patterns of these genes through RT-qPCR were similar to those of NGS RNAseq analysis. The results of this study can be applied to other fish species and it is considered to be useful industrially.

• Journal of Marine Life Science
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• v.3 no.2
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• pp.59-66
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• 2018

The assessment of level of health of the tidal flats can be evaluate by health of organisms inhabit the tidal flats. It is possible to evaluate the precise health level of organisms inhabit the tidal flats using analysis of expression of biomarker genes. The purpose of this research is to evaluate the health of the tidal flats on the west coast using biomarker genes such as heat shock protein 70 (Hsp70), heat shock protein 90 (Hsp90), glutathione S-transferases (GST) and thioredoxin (TRX). These genes are stress, immune, and antioxidant related genes that can be used to look at the health of an organism through gene expression. In this study, we collected manila clam (Ruditapes philippinarum) in 8 analysis areas on the west coast. Expression of the genes was analyzed by RT-qPCR method. Results showed that, the expression of Hsp70, Hsp90, GST and TRX genes were differentially expressed in the 8 analysis areas. In particular, the expression of Hsp90 and GST or the expression of Hsp70 and TRX were similar. This means that there is a substance that reacts specifically to each gene. Therefore, I think suggest that the based on the results of physicochemical analysis, it can be selected genes suitable for analysis. These results suggest that Hsp70, Hsp90, GST and TRX were played roles in biomarker for assessment of the health of tidal flats.