• 한국초지조사료학회지
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• 제17권4호
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• pp.379-386
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• 1997

대만산 배추 (Brassica campestris M.)로부터 분리한 BcHSPI7.6 cDNA(내열성 유전자)를 pBKSl-l vector에 subcloning하므로서, NPTII 유전자와 P35S-HSPI7.6 cDNA를 가지는 pBKH4 재조합 플라스미드를 제작하였다. 이들 플라스미드를 갖는 A. tumefaciens LBA4404로서 담배잎 단편을 24시간 동안 공배양하므로서 감염시켰으며, 이들 유전자로 형질전환된 shoot는 $100\;{\mu\textrm{g}}/ml$의 가나마이신을 첨가한 MS-n/B 배지에서 선발하였다. 담배((Nicotiana. tabacum)의 열에 대한 치사온도는 $50^{\circ}$에서 15분 이상이었으며, BcHSPI7.6 cDNA를 갖는 형질전환된 식물체는 이 온도에서 내열성을 나타내었다. 식울체의 형질전환 여부는 ${\alpha}^{_32}P$로 표지한 BcHSPI7.6 cDNA 단편을 probe로 이용해서 Southern blot hybridization을 실시하므로서 확인하였다. BcHSPI7.6 cDNA의 발현정도는 Northern blot 분석과 이중면역확산 방법으로 확인하였다. 본 연구에서, 담배에 도입한 BcHSPI7.6 cDNA는 내열성과 관련이 있는 유전자로서, HSPI7.6 단백질은 식물체를 열에 의한 손상으로부터 방지해 주는 protector 역할을 하는 것 으로 사료된다.

• 한국자기공명학회논문지
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• 제17권1호
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• pp.54-58
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• 2013

SAV0506 is an 87 residue hypothetical protein from Staphylococcus aureus strain Mu50 and also predicted to have similar function to ribosome associated heat shock protein, Hsp 15. Hsp15 is thought to be involved in the repair mechanism of erroneously produced 50S ribosome subunit. In this report, we present the sequence specific backbone resonance assignment of SAV0506. About 82.5% of all resonances could be assigned unambiguously. By analyzing deviations of the $C{\alpha}$ and $C{\beta}$ chemical shift values, we could predict the secondary structure of SAV0506. This study is an essential step towards the structural characterization of SAV0506.

• Clinical and Experimental Reproductive Medicine
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• 제26권1호
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• pp.103-109
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• 1999

Objective: Heat shock protein 70-2 (Hsp70-2) gene knockout mice are found to have premeiotic arrest at the primary spermatocyte stage with a complete absence of spermatids and spermatozoa. This observation led to the hypothesis that hspA2 may be disrupted in human testes with abnormal spermatogenesis. To test this hypothesis, we studied the mRNA expression of hspA2 in infertile men with azoospermia. Design: The mRNA expression were analyzed by competitive RT-PCR among testes with normal spermatogenesis, pachytene spermatocyte arrest, and sertoli-cell only syndrome. Materials and methods: Testicular biopsy was performed in men with azoospermia (n=15). Specimens were subdivided into three groups: (group 1) normal spermatogenesis (n=5), (group 2) spermatocyte arrest (n=5), (group 3) Sertoli-cell only syndrome (n=5). Total RNA was extracted by Trizol reagent. Total extracted RNA was reverse transcribed into cDNA and amplified by PCR using specific primers for hspA2 target cDNAs. A competitive cDNA fragment was constructed by deleting a defined fragment from the target cDNA sequence, and then coamplified with the target cDNA for competitive PCR. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene was used as an internal control. Results: On Competitive RT-PCR analyses for hspA2 mRNA, significant amount of hspA2 expression was observed in group 1, whereas a constitutively low level of hspA2 was expressed in groups 2 and 3. Conclusion(s): The study demonstrates that the hspA2 gene expression is down-regulated in human testes with abnormal spermatogenesis, which in turn suggests that hspA2 gene may play a specific role during meiosis in human testes.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• 제4권2호
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• pp.181-190
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• 2001

목적: 위장관 증상을 갖는 HSP 환아에서 장관초음파소견과 위내시경검사소견을 규명하여 피부자반증없이 위장관 증상만 있는 환아에서 HSP 조기진단에 장관초음파검사와 위내시경검사의 유용성을 알아보고자 본 조사를 시행하였다. 방법: 1999년 1월부터 2001년 4월까지 동강병원 소아과에 입원한 위장관 증상을 동반한 HSP 환아85명 88례를 대상으로 임상증상, 장관초음파소견과 위내시경소견을 후향적으로 조사하였고 동일한 환아에게서 장관초음파검사와 위내시경검사를 동시에 시행한 4례의 환아에서 두 검사의 연관성을 찾아보았다. 결과: 1) 피부자반증이 88례(100%) 모두에서 나타났고 관절통 및 관절증상이 64례(73%), 위장관 증상이 52례(59%), 신장침범소견이 15례(17%), 음낭침범소견이 3례(3%)에서 나타났다. 2) 위장관 증상은 복통이 52례(100%)에서 나타났고 대변잠혈양성반응이 28례(54%), 구역 및 구토가 17례(33%), 복부압통이 17례(33%), 혈변 혹은 흑색변이 12례(23%), 설사 6례(12%), 토혈 4례(8%), 복부반사통과 복부경직이 각각 1례(2%)순이었다. 3) 위장관 증상을 갖는 52례 중 27례에서 복부초음파검사를 시행하였는데 22례(81%)에서 이상소견이 나타났다. 십이지장과 공장벽의 비후소견이 16례(73%)로서 가장 많았고 복수 8례(36%), 장간막임파선종창 8례(36%), 장폐색증 4례(18%), 이상담낭소견 1례(5%)순이었다. 위장관 증상을 갖지 않는 36례의 HSP 환아 중 3례에서 실시한 복부초음파검사는 모두 정상 소견으로 나타났다. 4) 5례의 내시경 소견은 전례에서 위소견은 정상이었고 십이지장에 다발성의 미란성 출혈성 십이지장염의 소견을 보였는데 구부보다 하행십이지장에 더 심했으며 1례에서는 구부소견은 정상이었고 하행십이지장에서만 동일한 소견을 보였다. 이중 2례에서 시행한 내시경적 생검절편의 병리조직학적 소견은 급성 염증반응과 국소미란 및 출혈소견을 보였다. 5) 4례에서 내시경검사와 복부초음파검사를 같이 시행하였는데 정상소견을 보인 1례를 제외하고는 십이지장과 공장벽의 비후소견의 초음파소견을 보여 미란성 궤양성 출혈성 십이지장염의 내시경소견이 초음파검사상 십이지장벽의 비후소견으로 나타나는 것으로 생각되었다. 결론: 위장관 증상을 동반하는 HSP 환아에서 복부초음파검사는 HSP의 위장관변화를 발견하고 경과관찰을 보는 간편하고 비침습적인 유용한 검사방법이며, HSP 환아에서 내시경검사의 적절한 적응증은 아직 확립되어 있지 않으나 하행십이지장의 출혈성 미란성 십이지장염이 일관된 소견으로 생각된다. 피부자반증이 없이 급성 상복부통증을 동반하는 위장관 증상, 대변잠혈 양성반응이 있을 경우, 복부초음파검사상 십이지장과 공장벽의 비후소견이 보이거나 혹은 위내시경상 출혈성 미란성 십이지장염의 소견이 보일 경우 pathognomonic하지는 않지만 HSP의 가능성을 항상 염두에 두고 다른 질환이 배제되면 부신피질호르몬 투여를 고려해보는 것이 좋을 것으로 생각된다.

• 한국자기공명학회논문지
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• 제16권1호
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• pp.22-33
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• 2012

Tid1, belonging to the Hsp40/DnaJ family of proteins, functions as a cochaperone of cytosolic and mitochondrial Hsp70 proteins. In particular, the N-terminal J-domain of Tid1 (Tid1-JD) constitutes the major binding sites for proteinprotein interactions with client proteins, including p53, as well as its partner chaperone, Hsp70. In the present study, soluble, recombinant protein of Tid1-JD could be obtained by using the pCold vector system, and backbone NMR assignments were completed using the isotope $[^{13}C/^{15}N]$-enriched protein. Far-UV CD result implied that Tid1-JD is an ${\alpha}$-helical protein and the secondary structure determined using chemical shift data sets indentified four ${\alpha}$-helices with a loop region containing the HPD (conserved tripeptide of His, Pro and Asp) motif. Additionally, NMR spectra under different conditions implied that the HPD motif, which is a critical region for protein-protein interactions of Tid1-JD, would possess dynamic properties.

• Asian Pacific Journal of Cancer Prevention
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• 제16권15호
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• pp.6491-6499
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• 2015

Background: Red-fleshed sweet orange juice (ROJ) comes from a new variety of citrus cultivated in Brazil that contains high levels of ${\beta}$-carotene and lycopene, and similar amounts of hesperidin (HSP) and nutrients, equivalently to blond orange juice (BOJ). Such bioactive compounds are associated with chemopreventive actions in several cancer cell lines. The purpose of this study was to examine the cytotoxicity, cell cycle, apoptosis, and cytokine secretion after BOJ, ROJ, and HSP treatment of a novel T acute lymphoblastic leukemia cell line, Loucy. Materials and Methods: Loucy cells were incubated for 24-h with BOJ, ROJ, and HSP, and the viability was measured using trypan blue. Cell cycling and apoptosis were assessed by propidium iodide (PI) and annexin V-FITC/PI flow cytometry, respectively. Secretion of cytokines $IL-1{\alpha}$, $IL1-{\beta}$, IL-2, IL-4, IL-6, IL-10, IL-17A, $IFN{\gamma}$, $TNF{\alpha}$, $TGF{\beta}$, $MIP{\alpha}$, and $MIP{\beta}$ was determined by ELISA array. Results: BOJ and ROJ treatments promoted Loucy cell cytotoxicity. Additionally, BOJ induced cell cycle arrest in the G0/G1 phase, and decreased the cell accumulation in the G2/M. ROJ decreased only the G0/G1 fraction, while HSP did not change the cell cycle. BOJ led to apoptosis in a different fashion of ROJ, while the first treatment induced apoptosis by increase of late apoptosis and primary necrotic fractions, the second increased early and late apoptosis, and primary necrotic fraction compared to positive controls. HSP had no effect on apoptosis. IL-6 and IL-10 were abrogated by all treatments. Conclusions: Taking together, these results suggest potential chemopreventive effects of BOJ and ROJ on Loucy cells.

• 한국해양생명과학회지
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• 제2권1호
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• pp.12-19
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• 2017

급격한 수온의 변화는 어류의 생리학적인 측면에서 스트레스를 유발한다. 본 연구에서는 넙치(Paralichthys olivaceus)로부터 각 수온별(9, 12, 15, 18 및 21℃) 조건에 따라 24 및 48시간 동안 노출시킨 후에, 혈액생리학적 분석, 스트레스 단백질로 알려진 Hsp70 mRNA 발현 및 산소 소비량을 조사하였다. 혈액학적 분석에서 hematocrit (Ht) 및 hemoglobin (Hb), 혈장 코티졸 및 글루코스의 변화, aspartate aminotransferase (AST) 및 alanine aminotransferase (ALT), NH₃, 삼투질농도(osmolality) 및 총단백질(total protein, TP)은 9℃ 및 12℃에서 다른 수온별 실험구에 비해 대부분의 항목에서 유의적인 차이를 보였다. Hsp70 mRNA 발현은 9℃ 및 12℃에서 다른 실험구에 비해 높은 발현량을 확인하였고, 산소소비량은 9℃ 및 12℃에서 21℃에 비해 낮았다. 이러한 결과는 넙치 종자의 장거리 수송을 위한 수온자료로 활용할 수 있다.

• IMMUNE NETWORK
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• 제4권1호
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• pp.7-15
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• 2004

Background: The heat shock proteins (HSPs) play an important role in cellular protection mechanisms against physical or chemical stresses. In this study scFv antibodies specific for human HSP70.1 were isolated from a semi-synthetic human scFv library with the ultimate goal of developing anti-HSP70.1 intracellular antibody (intrabody) that may offer an attractive alternative to gene targeting to study the function of the protein in cells. Methods: A semi-synthetic human scFv display library ($5{\times}10^{8}$ size) was constructed using pCANTAB-5E vector and the selection of the library against bacterially expressed recombinant human HSP70.1 was attempted by panning. Results: Three positive clones specific for recombinant HSP70.1 were identified. All three clones used $V_{H}$ subgroup III. On the other hand, $V_{L}$ of two clones belonged to the kappa light chain subgroup I, but the other utilized $V_{k}$ subgroup IV Interestingly, these scFv molecules specifically reacted to the recombinant HSP70.1, yet failed to recognize native HSP70 induced in U937 human monocytic cells by heat treatment. Conclusion: Our results indicated that affinity selection of an scFv phage display library using recombinant antigens produced in E. coli might not guarantee the isolation of scFv antibody molecules specific for a native form of the antigen. Therefore, the source of target antigens needs to be chosen carefully in order to isolate biofunctional antibody molecules.

• Childhood Kidney Diseases
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• 제10권2호
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• pp.125-131
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• 2006

Purpose : High interleukin-1 beta(IL-$1{\beta}$) expression in the skin biopsy specimens of patients with Henoch-$Sch\ddot{o}nlein$ Purpura(HSP) has been observed. We examined IL-$1{\beta}$ gene polymorphism in patients with HSP. The purpose of this study is to examine the relationship between IL-$1{\beta}$ gene polymorphism and renal involvement in HSP. Methods : Patients from mideast Korea with HSP were studied. All patients had at least 6 months of follow up. Patients and ethnically matched controls were genotyped for IL-$1{\beta}$ gene polymorphism by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP). Results : Thirty-four patients(all younger than 15 years old) who had been diagnosed with HSP and 27 controls were examined. No allele or genotype differences between the HSP and control groups were observed. No significant association between the carriage of IL-$1{\beta}$(-511) T allele and renal involvement(P=0.525, OR:1.417, CI:0.545-3.686) was found. Conclusion : In unselected patients with HSP, carriage of IL-$1{\beta}$(-511) T allele does not appear to influence renal involvement.

• The Korean Journal of Physiology and Pharmacology
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• 제4권4호
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• pp.319-324
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• 2000

The causal relationship between heat shock protein (HSP) and second window of cardioprotective effect is still undetermined. In the present study, we assessed whether HSP-producing substances, amphetamine and ketamine, afforded protection against reperfusion-induced ventricular fibrillation (VF) and these protective effect remained after the inhibition of HSP72 production by quercetin, a mitochondrial ATPase inhibitor. Adult mongreal male cats $(n=60,\;2.5{\sim}4\;kg)$ were used in this study. Experimental animals were divided into five groups; control group (n=15), amphetamine ('A', n=11) group, ketamine ('K', n=9) group, amphetamine-ketamine ('AK', n=16) group and amphetamine-ketamine-quercetin ('AKQ', n=9) group. Twenty-four hours after the drug treatment, an episode of 20-min coronary artery occlusion was followed by 10-min reperfusion. The incidence of reperfusion-induced VF in the AK and AKQ groups was significantly lower than that in control group (p＜0.01). After the ischemia/reperfusion procedure, western blot analysis of HSP72 expression in the myocardial tissues resected from each group was performed. HSP72 production in the AK group was marked, whereas HSP72 was not detected in the AKQ and control groups. These results suggest that the suppressive effect against reperfusion-induced VF induced by amphetamine and ketamine is not mediated by myocardial HSP72 production but by other mechanisms.