• BMB Reports
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• 제40권4호
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• pp.554-561
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• 2007

Shen and colleagues (Lin et al., 2004) have recently shown that overexpression of the Drosophila DNA methyltransferase 2 isoform C, dDnmt2c, extended life span of fruit flies, probably due to increased expression of small heat shock proteins such as Hsp22 or Hsp26. Here, I demonstrate with immunoprecipitations that overexpressed dDnmt2c interacts with endogenous Hsp70 protein in vivo in S2 cells. However, its C-terminal half, dDnmt2c(178-345) forms approximately 10-fold more Hsp70-containing protein complexe than wild-type dDnmt2c. Overexpressed dDnmt2c(178-345) but not the full length dDnmt2c is able to increase endogenous mRNA levels of the small heat shock proteins, Hsp26 and Hsp22. I provide evidence that dDnmt2c(178-345) increases Hsp26 promoter activity via two heat shock elements, HSE6 and HSE7. Simultaneously overexpressed Hsp40 or a dominant negative form of heat shock factor abrogates the dDnmt2c(178-345)-dependent increase in Hsp26 transcription. The data support a model in which the activation of heat shock factor normally found as an inactive monomer bound to chaperones is linked to the overexpressed C-terminus of dDnmt2c. Despite the differences observed in flies and S2 cells, these findings provide a possible explanation for the extended lifespan in dDnmt2c-overexpressing flies with increased levels of small heat shock proteins.

• 국제물리치료학회지
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• 제2권2호
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• pp.281-287
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• 2011

This research was attempted to seek for a positive approach within the framework of physical therapy instead of the drug treatment in the past, with regard to the ischemic brain injury in the early stage. Accordingly, the aim of this research is to observe the change of HSP27 and HSP70, the genes that are expressed in the early stage of brain injury and to investigate the effects of needle electrode electrical stimulation(NEES), upon applying NEES after ischemia. The experimental method is to give rise to global ischemia and apply NEES to 27 SD-Pat rats with the particulars of being eight-week-old, male, around 300g, and adapted to laboratory environment for more than a week, and divide them into three groups, that is, GV20 NEES group(n=9), L14 NEES group(n=9), no applied NEES global ischemia(GI) group(n=9), and then observe their changes of HSP27 and HSP70 at the time lapse of 6, 9 hr and 12 hr after ischemia, using immunohistochemistry methods. Upon observing through the immunohistochemistry method, it was noticed that there was a significant difference between the GV20 NEES group and the L14 NEES group as for HSP27 and there were significant differences among all groups as for HSP70(p<.05). Accordingly, it is supposed that the application of NEES after the outbreak of cerebral ischemia delay the apoptosis in the early ischemic part of forebrain or protect neurons against apoptosis.

• Journal of Periodontal and Implant Science
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• 제37권1호
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• pp.11-21
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• 2007

Heat shock protein (HSP) is one of cellular protein commonly present in major periodontopathogenic bacteria as well as mammalian cells. The protein may play a role in the immunopathogenesis by modulating autoimmune reaction due to its high level of sequence homology between bacteria and human counterpart. Hence, identifying immunodomiant epitope of bacteria HSP that is cross-reactive to periodontopathogenic bacteria with a specificity to human HSP may comprise a critical strategy for development of a periodontal vaccine. The present study was performed to establish clones producing monoclonal antibody reactive to Porphyromonas gingivalis (p. gingivalis) HSP with a specificity to human HSP. 4 different hybridomas were cloned producing monoclonal IgG antibodies to P, gingivalis HSP and evaluated for their reactivity and specificity to other periodontopathogenic bacteria as well as to human HSP. These four monoclonal antibodies reacted with p. gingivalis HSP only with specificities to other bacteria tested and human HSP as well. The antigenic epitopes producing the 4 monoclonal antibody may be potentially developed as vaccine candidates. Further investigations are under way to identify more clones producing monoclonal antibodies reactive to P, gingivalis HSP and to other periodontopathogenic bacteria as well, while maintaining specificities to human counterpart.

• 한국동물학회지
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• 제30권3호
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• pp.311-323
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• 1987

생쥐의 섬 유아세포(MEP)와 종양세포(SCK)를 이용하여 정상세포와 종양세포 사이에 열 감수성의 차이가 있는지의 여부 및 환경의 pH가 이 세포들의 열감수성과 heat shock protein(HSP) 합성에 미치는 영향을 생존곡선과 HSP합성 kinetics등을 써서 검토하였다. MEF와 SCK 세포를 정상 pH(7.4) 또는 산성 pH(6.7)에서 42"C에서 2시간 온열처리 후 3일간에 걸쳐 생존을을 비교해 븐 결과, ME선와 SCK세포 사이에 생득적 열강수성의 차이는 없었고 산성 P광에서는 세포의 종류에 관계없이 열감수성 이 증감되었다. 온열처리의 결과 유도되는 내일성이 conditioning Leat의 크기와 어떤 관계가 있는지를 보기 위해서 45"C에서 5분 또는 20분을 주어본 결과 체은 conditioning heat를 주었을 때 내일성이 신속히 그리고 높은 수준으로 발생하였고, 이러한 열 감수성의 kinetics는 HSP의 합성 kinetics와 잘 일치하였다. 단백질, 특히 HSP 합성에 미치는 PH의 영 향을 알아보기 위해서 46"C에서 6분간의 heat shock를 주어 본 바 전반적인 단백질 및 major HSP의 합성양상에는 별로 차이를 보이지 않았다. 그러나 SCK 세포에 43"C에서 30분의 온열처리를 주고 새로 합성되는 HSPSP의 kinetics를 검토해 본 결과 정상 P반에서는 0-5시간에 합성이 일어나나 산성 PH에서는 3-9시간에 합성이 일어나서 몇시간의 합성지연이 관찰되었다. 아울러 HSP68, HSPTC, HSP87을 Peptidemapping하여 본 결과 HSP68과 HSP70 은 유사한 peptide fragment pattern을 보여 amino acid sequence는 유사하고 기능도 같을 것으로 추론되었으나 HSP87은 전혀 다른 pattern을 보였다. 전혀 다른 pattern을 보였다.

• 한국동물학회지
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• 제39권2호
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• pp.123-131
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• 1996

In this study, we examined the expression of heat shock proteins (HSPs) in fish cell line CHSE-2lnl and human HeLa cells exposed to heat shock. In fish CHSE-214 cells HSP70 was the major polvpeptide induced by an elevated temperature or an amino acid analog, while in HeLa cells HSP90 as well as HSP70 were prominently enhanced in response to these stresses. Pretreatment of actinomvcin D prior to heat shock completely inhibited the induction of fish HSP70, indicating the transcriptional regulation of fish HSP70 gene expression. In HeLa and CHSE-214 cells either recovering from heat shock or experiencing prolonged heat shock, attenuation in the HSP90 a'nd HSP70 induction occurred but both induction and repression of HSP70 synthesis appear 19 precede those of HSP90. Moreover, attenuation did not occur in the syntheses of 40 kDa and 42 kOto proteins which were only induced in CHSE-214 cells. The enhanced syntheses of these he proteins continued as long as CHSE-214 cells were Siven heat shock. These results suggest that down-regulation of HSP syntheses during prolonged heat shock may be controlled by several different. as vet undefined, mechanisms.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제23권2호
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• pp.124-136
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• 2001

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 HSP70 and HSP47 expression was assessed in the DMBA induced oral carcinogenesis in hamster. Golden Syrian hamsters which were 3 months-age and $90{\sim}120g$ 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 race 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. HSP47 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.

• 한국환경성돌연변이발암원학회지
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• 제25권2호
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• pp.51-59
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• 2005

Overexpression of HSP25 delayed cell growth, increased the level of $p21^{waf}$, reduced the levels of cyclin D1, cylcin A and cdc2, and induced radioresistance in L929 cells. We demonstrated that extracellular regulated kinase (ERK) and MAP kinase/ERK kinase (MEK) expressions as well as their activation (phospho-forms) were inhibited by hsp25 overexpression. To confirm the relationship between ERK1/2 and hsp25-mediated radioresistance, ERK1 or ERK2 cDNA was transiently transfected into the hsp25 overexpressed cells and their radioresistance was examined. HSP25-mediated radioresistance was abolished by overexpression of ERK2, but not by overexpression of ERK1. Alteration of cell cycle distribution and cell cycle related protein expressions (cyclin D, cyclin A and cdc2) by hsp25 overexpression were also recovered by ERK2 cDNA transfection. Increase in Bc1-2 protein by hsp25 gene transfection was also reduced by subsequent ERK2 cDNA-transfection. In addition, HSP25 overexpression reduced reactive oxygen species (ROS) and increased expression of manganese superoxide dismutase (MnSOD) gene. Increased activation of NF-kB (IkB degradation) was also found in hsp25-overexpressed cells. Moreover, transfection of hsp25 antisense gene abrogated all the HSP25-mediated phenomena. To further elucidate the exact relationship between MnSOD induction and NF-kB activation, dominant negative $I-kB\alpha(I-kB\alpha-DN)$ construction was transfected to HSP25 overexpressed cells. $I-kB\alpha-DN$ inhibited HSP25 mediated MnSOD gene expression. In addition, HSP25 mediated radioresistance was blocked by $I-kB\alpha-DN$ transfection. Blockage of MnSOD with antisense oligonucleotides in HSP25 overexpressed cells, prevented apoptosis and returned the ERK1/2 activation to the control level. From the above results, we suggest for the first time that reduced oxidative damage by HSP25 was due to MnSOD-mediated down regulation of ERK1/2.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제20권4호
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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.

• 한국수정란이식학회지
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• 제20권3호
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• pp.289-295
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• 2005

본 연구는 돼지에서 스트레스 관련 HSP70 유전자의 다형성과 IVF 수정란의 배발달간의 관련성을 조사하기 위하여 수행되었으며 그 결과는 다음과 같다. HSP70-K1, -K3 및 -K4의 PCR 산물로부터 SSCP 다형성은 각각 다르게 확인되었다. 체외수정란의 분할율 시험에서 HSP70 K1-AA($73.1\%$) 및 K1-AB($62.3\%$) 난할율은 HSP70 K1-BB($49.3\%$)보다 유의적으로 높게 나타났다(p<0.05). 또한 HSP70 K3-AA($72.4\%$) and K3-AB($62.2\%$)형도 HSP70 K3-BB($49.1\%$)형보다 유의적으로 높았다(p<0.05). 돼지품종들과 정자의 HSP70 유전자형에 따른 2-cell 단계까지의 체외수정란 배발달은 유의적 차이를 보였다. Landrace(28.8)와 Duroc(29.8)에서 2세포기까지 발달된 수정란의 수는 Yo.kshire(10.9) 보다 유의적으로 높았다(p<0.05). 또한 HSP70 K4-AB(29.6)형의 2세포기 단계까지의 배발달 수정란 수는 HSP70 K4-AA(10.6)형보다 유의적으로 높았다(p<0.05). 그러나 돼지 품종 및 HSP70 유전자형에 따른 배반포기까지 수정란 배발달 정도는 유의적 차이는 없었다. 이러한 결과들은 돼지 체외수정란의 초기 배발달은 HSP70 유전자형과 품종에 따라서 영향을 받는 것으로 확인되었다.

• 식물조직배양학회지
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• 제27권1호
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• pp.13-18
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• 2000

고등식물에 있어서 엽록체에 존재하는 저 분자량 HSP의 기능을 밝히기 위하여 담배 (Nicotina tabacum cv. Petit Havana SRI)로부터 분리한 cDNA (NtHSP21)를 도입한 형질전환 담배 식물체를 재생하였다. 상온에서의 발현량이 서로 다른 5개의 순계 형질전환 식물체를 선발하였다 상온에서 발현된 엽록체 small HSP가 식물의 고온내성에 미치는 영향을 조사하기 위하여 기내에서 생장시킨 유식물을 52$^{\circ}C$에서 45분간 열처리한 후 생장온도에서의 변화를 조사하였다. 그 결과 wild-type식물의 경우 1주일 이내에 모두 고사하였으나 형질전환 식물체의 약 70%는 생존하였다. 또한 이러한 고온내 성은 상온에서 발현된 단백질의 양에 비례하여 증감하였다. 따라서 엽록체에 존재하는 small HSP가 식물의 고온내성 획득에 있어서 중요한 역할을 담당할 것으로 사료된다.