• Molecules and Cells
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• v.37 no.11
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• pp.833-840
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• 2014

Cullin4-RING ubiquitin ligase (CRL4) is a family of multi-subunit E3 ligases. To investigate the possible involvement of CRL4 in heat stress response, we screened T-DNA insertion mutants of putative CRL4 substrate receptors that exhibited altered patterns in response to heat stress. One of the mutants exhibited heat stress tolerance and was named heat stress tolerant DWD1 (htd1). Introduction of HTD1 gene into htd1-1 led to recovery of heat sensitivity to the wild type level, confirming that the decrease of HTD1 transcripts resulted in heat tolerance. Therefore, HTD1 plays a negative role in thermotolerance in Arabidopsis. Additionally, HTD1 directly interacted with DDB1a in yeast two-hybrid assays and associated with DDB1b in vivo, supporting that it could be a part of a CRL4 complex. Various heat-inducible genes such as HSP14.7, HSP21, At2g03020 and WRKY28 were hyper-induced in htd1-1, indicating that HTD1 could function as a negative regulator for the expression of such genes and that these genes might contribute to thermotolerance of htd1-1, at least in part. HTD1 was associated with HSP90-1, a crucial regulator of thermotolerance, in vivo, even though the decrease of HTD1 did not affect the accumulation pattern of HSP90-1 in Arabidopsis. These findings indicate that a negative role of HTD1 in thermotolerance might be achieved through its association with HSP90-1, possibly by disturbing the action of HSP90-1, not by the degradation of HSP90-1. This study will serve as an important step toward understanding of the functional connection between CRL4-mediated processes and plant heat stress signaling.

• Journal of Microbiology and Biotechnology
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• v.28 no.4
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• pp.542-550
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• 2018

Heat shock protein 90 (Hsp90) is treated as a molecular therapeutic target for the prevention and treatment of cancer. Geldanamycin (GA) was the first identified natural Hsp90 inhibitor, but hepatotoxicity has limited its clinical application. Nevertheless, a new GA analog (WK-88-1) with the non-benzoquinone skeleton, obtained from genetically engineered Streptomyces hygroscopicus, was found to have anticancer activity against two human breast cancer cell lines. WK-88-1 produced concentration-dependent inhibition of cell proliferation, cell cycle arrest, and apoptosis in estrogen receptor (ER)-positive MCF-7 and ER-negative MDA-MB-231 cell lines. Detailed analysis showed that WK-88-1 downregulated some key cell cycle molecules (CDK1 and cyclin B1) and lead to $G_2/M$ cell cycle arrest. Further studies also showed that WK-88-1 could induce human breast cancer cell apoptosis by downregulating Hsp90 client proteins (Akt, p-Akt, IKK, c-Raf, and Bcl-2), decreasing the ATP level, increasing reactive oxygen species production, and lowering the mitochondrial membrane potential. Meanwhile, we discovered that WK-88-1 significantly decreased the levels of Her-2 and $ER-{\alpha}$ in MCF-7 cells but not in MDA-MB-231 cells. In addition, WK-88-1 significantly increased caspase-3, -8, and -9 activities and the cleavage of PARP in a concentration-dependent manner (with the exception of caspase-3 and PARP in MCF-7 cells). Taken together, our preliminary results suggest that WK-88-1 has the potential to play a role in breast cancer therapy.

• Korean Journal of Poultry Science
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• v.47 no.1
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• pp.29-37
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• 2020

This study was performed to investigate the effect of mixed rearing of male and female chickens on the stress response in Korean native chickens. To identify the degree of the stress response, heterophil-lymphocyte ratio (H/L ratio), heat shock protein genes (HSPs) expression, and intracellular nuclear DNA damage rate were analyzed before and after the mixed rearing of male and female chickens. The results showed that the H/L ratio of chickens after mixing males and females was more than thrice as higher than before mixing (P<0.001), but the differences between males and females were not significant. HSP-70, HSP90-α, and HSP90-β expression levels were 2.5 to 3.4 times higher after mixing male and female chickens, compared to before mixing (P<0.01). In the mixed rearing of male and female chickens, the increase in HSPs expression in females was higher than in males. Comet indicators in intracellular DNA damage rate analysis showed a significant increase after mixing male and female chickens compared to before mixing (P<0.001). However, there was no significant difference between males and females with respect to DNA damage rate. Taken together, these results suggest that male and female mixed rearing acts as a strong external stressor in both male and female chickens.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.17
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• pp.7043-7048
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• 2014

Inhibition of heat shock protein 90 (Hsp90) leads to inappropriate processing of proteins involved in DNA damage repair pathways after DNA damage and may enhance tumor cell radio- and chemotherapy sensitivity. To investigate the potentiation of antitumor efficacy of lidamycin (LDM), an enediyne agent by the Hsp90 inhibitorgeldanamycin (GDM), and possible mechanisms, we have determined effects on ovarian cancer SKOV-3, hepatoma Bel-7402 and HepG2 cells by MTT assay, apoptosis assay, and cell cycle analysis. DNA damage was investigated with H2AX C-terminal phosphorylation (${\gamma}H2AX$) assays. We found that GDM synergistically sensitized SKOV-3 and Bel-7402 cells to the enediyne LDM, and this was accompanied by increased apoptosis. GDM pretreatment resulted in a greater LDM-induced DNA damage and reduced DNA repair as compared with LDM alone. However, in HepG2 cells GDM did not show significant sensitizing effects both in MTT assay and in DNA damage repair. Abrogation of LDM-induced $G_2/M$ arrest by GDM was found in SKOV-3 but not in HepG2 cells. Furthermore, the expression of ATM, related to DNA damage repair responses, was also decreased by GDM in SKOV-3 and Bel-7402 cells but not in HepG2 cells. These results demonstrate that Hsp90 inhibitors may potentiate the antitumor efficacy of LDM, possibly by reducing the repair of LDM-induced DNA damage.

• Korean Journal of Acupuncture
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• v.31 no.4
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• pp.218-224
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• 2014

Objectives : Zizyphus jujuba fruits(ZJF), a traditional Korean medicine has various biological activities such as anti-inflammatory, anti-oxidative and neuro-protective effects. However, it is still unclear whether ZJF has any biological effect on anti-aging. In this study, we examined the effect of ZJF on lifespan and thermal stress in C. elegans. Methods and Results : ZJF water extracts were fermented for 7 days(F7-ZJF) and 14 days(F14-ZJF) by Laetiporus sulphureus to increase secondary metabolites such as aglycone of flavonoids and terpenoids. In the lifespan assay, ZJF water extracts and fermented ZJF were treated on the agar medium plate with age synchronized egg stage of C. elegans. Treatment of F7-ZJF-$200{\mu}g/mL$ with OP-50 E. coli and F14-ZJF-$200{\mu}g/mL$ with OP-50 E. coli significantly increased life span of C. elegans(N2) at thermal stress condition of $25^{\circ}C$. Moreover mRNA levels of lifespan associated HSP 16.1, HSP 70, and HSF-1 were increased at thermal stress condition of $25^{\circ}C$. However, in the equilibration temperature of $20^{\circ}C$ after stress condition of $35^{\circ}C$ for 2 hr, F-14-ZJF-$200{\mu}g/mL$ treatment decreased the levels of heat shock protein in hsp16.2/GFP C. elegans. Conclusions : Our study indicates that prolong role of fermented-ZJF in C. elegans is mediated by control HSPs production.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.36 no.5
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• pp.346-352
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• 2010

Introduction: Heat shock protein70 (HSP70) is a highly conserved family of proteins produced after a variety of stresses. Many studies reported that the overexpression of HSP70 can improve the prognosis of the patients with sepsis through a reduction of the nitric oxide concentration. However, these results only revealed the effect of HSP70 and nitric oxide. No studies have examined the relationship between HSP70 and nitric oxide. The aim of this study was to evaluate the effect of the overexpression of HSP70 on the expression of inducible nitric oxide synthase and the nitric oxide concentration. In addition, the mechanism of the relationship of HSP70 and inducible nitric oxide synthase (iNOS) in sepsis was examined. Materials and Methods: The experiments were performed on male sprague-dawley rats. Sepsis was induced by a cecal ligation and puncture (CLP). Glutamine (GLN) or saline was administered 1 hour after the initiation of sepsis. Serum and lung tissues were acquired from the rats 12 hours or 24 hours after the initiation of sepsis. The nitric oxide concentration, the expression of HSP70 in lung, and the gene expression of iNOS in lung were analyzed. The three groups, sham operation, CLP and CLP+GLN, were compared. Results: Compared to the other groups, in CLP+GLN, GLN administered after the initiation of sepsis enhanced the expression of HSP70 in the lung at 12 hours ($47.19{\pm}10.04$ vs. $33.22{\pm}8.28$, P=0.025) and 24 hours ($47.06{\pm}10.60$ vs. $31.90{\pm}4.83$, P=0.004). In CLP+GLN, GLN attenuated the expression of iNOS messenger RNA (mRNA) in the lung at 12 hours ($5,513.73{\pm}1,051.60$ vs. $4,167.17{\pm}951.59$, P=0.025) and 24 hours ($18,740.27{\pm}8,241.20$ vs. $9,437.65{\pm}2,521.07$, P=0.016), and reduced the concentration of nitric oxide in the serum at 12 hours ($0.86{\pm}0.48$ vs. $3.82{\pm}2.53$, P=0.016) and 24 hours ($0.39{\pm}0.25$ vs. $1.85{\pm}1.70$, P=0.025). Conclusion: The overexpression of HSP70 induced by the administration of GLN in sepsis attenuates the expression of the iNOS gene but reduces the nitric oxide concentration.

• Herbal Formula Science
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• v.21 no.2
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• pp.144-157
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• 2013

Objectives : The aim of this study was to evaluate the efficacy of Hwangnyeonhaedok-tang (HH) and Geongangbuja-tang (GB) on the plasma hormone level in mice exposed to cold stress. HH and GB are the representative prescriptions of cold and hot property, respectively. Methods : We established cold condition by confining ICR mice to a $4^{\circ}C$ cage for 24 hours, ICR mice were given a HH (100, 300, 1000 mg/kg) or GB (100, 300, 1000 mg/kg) extract orally twice a day for three consecutive days. From the second day, they were given cold stress ($4^{\circ}C$) for twenty four hours. To measure the plasma corticosterone, insulin, thyroxine, epinephrine and norepinephrine levels of mice, their blood samples were collected from cardiac puncture, immediately centrifuged at $4^{\circ}C$. The protein level of HSP70 and JNK was examined using western blot analysis in cortex and hypothalamus. Results : Oral administration of GB more significantly reduced plasma corticosterone level raised by cold stress than HH. Gardeniae Fructus (CJ), the constituent of HH, significantly increased the thyroxine level. Western blot analysis showed that cold stress-induced Heat shock protein 70 (HSP70) expression was increased by HH and GB, HH decreased JNK expression and GB increased JNK expression dose-depently in hypothalamus. Scutellariae Radix (HG), Zingiberis Rhizoma (GG) and Aconiti Tuber (BJ) decreased HSP70 in hypothalamus and GG, BJ decreased HSP70 in cortex as well. Conclusions : These results suggest Geongangbuja-tang (GB) is more effective for ameliorating the stress response caused by cold stress.

• Microbiology and Biotechnology Letters
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• v.25 no.2
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• pp.129-136
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• 1997

Helicobacter pylori is a spiral-shaped, microaerophilic human gastric pathogen causing chronic-active gastritis in association with duodenal ulcer and gastric cancer. To investigate the possibility of H. pylori outer membrane proteins (OMPS) as the oral vaccine antigens, sarcosine-insoluble outer membrane fraction has been prepared from H. pylori NCTC 11637. The major OMPs having apparent molecular masses of 62 kDa, 54 kDa and 33 kDa were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), which were identified as urease B subunit (UreB), heat shock protein (Hsp54 kDa) and urease A subunit (UreA), respectively. Minor protein bands of 57 kDa, 52 kDa, 40 kDa, 36 kDa and 31 kDa were also observed. The antigenicity of H. pylori OMPs and antigenic cross-reactivity among the strains were determined by immunoblot analysis using anti-H. pylori OMPs antisera or intestinal lavage solutions. The results showed that UreB, Hsp54 kDa, UreA and 40 kDa proteins vigorously stimulated mucosal immune response rather than systemic immunity. From this results, these proteins seemed to be useful as the antigen candidates for the oral vaccine. The immunoblotting results with surface proteins from eight isolated H. pylori strains were similar to that of H. pylori NCTC 11637. The IgA which had been arised from oral administration of H. pylori OMPs, was able to bind H. pylori whole-cells.

• Journal of Periodontal and Implant Science
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• v.40 no.2
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• pp.61-68
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• 2010

Purpose: The present study was performed to clarify the relationship between periodontal disease severity and selected immunological parameters consisting of serum IgG titer against periodontopathogenic bacteria, the expression of the helper T-cell cytokine by gingival mononuclear cells, and patients' immunoreactivity to cross-reactive heat shock protein (HSP) epitope peptide from P. gingivalis HSP60. Methods: Twenty-five patients with moderate periodontitis had their gingival connective tissue harvested of gingival mononuclear cells during an open flap debridement procedure and peripheral blood was drawn by venipuncture to collect serum. The mean level of interproximal alveolar bone was calculated to be used as an index for periodontal disease severity for a given patient. Each of selected immunologic parameters was subject to statistical management to seek their correlations with the severity of periodontal disease. Results: A significant correlation could not be identified between serum IgG titers against specific bacteria (Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, Actinobacillus actinomycetemcomitans, and Streptococcus mutans) and the severity of periodontal disease. Expression of interleukin (IL)-10 by gingival mononuclear cells was statistically significant in the group of patients who had higher levels of alveolar bone height. However, a similar correlation could not be demonstrated in cases for IL-4 or interferon-$\gamma$. Patients' serum reactivity to cross-reactive epitope peptide showed a significant correlation with the amount of alveolar bone. Conclusions: It was concluded that expression of IL-10 by gingival mononuclear cells and patients' sero-reactivity to the cross-reactive HSP peptide of P. gingivalis HSP60 were significantly correlated with alveolar bone height.

• 대한생식의학회:학술대회논문집
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• 2006.06a
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• pp.150.2-150.2
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• 2006