• BMB Reports
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• 제55권9호
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• pp.413-416
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• 2022

Ferroptosis is a type of programmed cell death distinct from apoptosis or necroptosis. Ferroptosis is well characterized by an iron-dependent accumulation of lipid peroxides and disruption of cellular membrane integrity. Many metabolic alterations can prevent or accelerate ferroptosis induction. Recent advances in analytical techniques of mass spectrometry have allowed high-throughput analysis of metabolites known to be critical for understanding ferroptosis regulatory metabolism. In this review, we introduce mass spectrometry-based analytical methods contributing to recent discovery of various metabolic pathways regulating ferroptosis, focusing on cysteine metabolism, antioxidant metabolism, and poly-unsaturated fatty acid metabolism.

• 산업기술연구
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• 제42권1호
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• pp.29-36
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• 2022

The CRISPR system has revolutionized gene editing field. Cas9-mediated gene editing such as Indel induction or HDR enable targeted gene disruption or precise correction of mutation. Moreover, CRISPR-based new editing tools have been developed such as base editors. In this review, we focus on gene editing in human pluripotent stem cells, which is principal technique for gene correction therapy and disease modeling. Pluripotent stem cell-specific drug YM155 enabled selection of target gene-edited pluripotent stem cells. Also, we discussed base editing for treatment of congenital retina disease. Adenine base editor delivery as RNP form provide an approach for genetic disease treatment with safe and precise in vivo gene correction.

• 대한한의학방제학회지
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• 제26권3호
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• pp.251-259
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• 2018

Objective : Kurarinone is one of the flavonoids isolated from Sophorae Radix with various biological activities including anti-microbial effect. In this study, we investigated the effects of Kurarinone on tert-butyl hydroperoxide (tBHP)-induced oxidative stress finally leading to apoptosis in human hepatoma cell line HepG2. Methods : To determine the effects on cell viability, the cells were exposed to tBHP ($100{\mu}mol/l$) after pretreatment with kurarinone (0.5 and $1{\mu}g/ml$). Cell viability was measured by MTT assay. To reveal the possible mechanism of cytoprotectivity of kurarinone, levels of reactive oxygen species, intracellular glutathione, mitochondrial membrane potential, and expression of caspase were examined. Results : tBHP-induced cell death was due to oxidative stress and the resulting apoptosis. Kurarinone dose-dependently protected cells from apoptosis when determined by MTT and TUNEL assay. Consistent with this observation, decreased expression of pro-caspase 3/9 protein by tBHP was restored by kurarinone. Kurarinone also showed anti-oxidative effects by inhibiting generation of ROS and depletion of GSH in tBHP-stimulated HepG2 cells. In addition, kurarinone significantly recovered disruption of mitochondrial membrane potential (MMP) as a start sign of hepatic apoptosis induced by oxidative stress. Conclusion : From these results, it was concluded that kurarinone protected tBHP-induced hepatotoxicity with anti-oxidative and anti-apoptotic activities. Our results suggest that kurarinone might be beneficial to hepatic disorders caused by oxidative stress.

• 대한미생물학회지
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• 제11권1호
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• pp.1-11
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• 1976

Escherichia coli(ATCC 11115)에 실험실등에서 상용하는 여러가지 항균제를 시간별로 처리, 그 변화양상을 전자현미경으로 관찰한 바 그 결과를 요약하면 다음과 같다. 1) 대조군은 3층의 단일막으로 형성된 세포벽에 둘러쌓여 있으며 세포질은 전자밀도가 낮은 nucleoid와 ribosme들이 산재하여 있음을 관찰할 수 있었다. 2) 70% ethanol용액 처리군은 핵물질을 관찰할 수 없었고 세포질은 세포 중앙부로 응집되어 있었으며 세포벽의 외부에서는 bleb 들을 관찰 할 수 있었다. 3) 3% $H_2O_2$ 용액 처리군은 세포내용물의 변화는 70% ethanol 처리군과 대동소이(大同小異)하였으나 세포벽에서는 심한 굴곡현상이 관찰되었다. 4) 5% lysol 용액처리군은 세포질 및 핵물질 부위가 완전히 구분되어 나타났으며 세포질내의 ribosome과립들은 시간이 경과할수록 그 응집현상이 심하였고 세포 외부에는 ribosome 양 과립들이 부착하고 있음이 관찰되었다. 5) 1% DDEGH 용액 처리군은 세포질의 응집 및 세포막과 세포벽이 뚜렷이 관찰되지 않았으며 세포외부에 세포내용물과 동일한 물질로 생각되는 물질이 부착되어 있음을 관찰할 수 있었다. 6) 고압멸균 처리군은 세포막 및 세포벽의 파괴, 탈락 및 세포내용물의 유출현상이 관찰되었다.

• Journal of Microbiology and Biotechnology
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• 제17권7호
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• pp.1162-1168
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• 2007

Although widely used as a host for recombinant protein production, Escherichia coli is unsuitable for massive screening of recombinant clones, owing to its poor secretion of proteins. A vector system containing T4 holin and T7 lysozyme genes under the control of the ptsG promoter derivative that is inducible in the absence of glucose was developed for programmed cell lysis of E. coli. Because E. coli harboring the vector grows well in the presence of glucose, but is lysed upon glucose exhaustion, the activity of the foreign gene expressed in E. coli can be monitored easily without an additional step for cell disruption after the foreign gene is expressed sufficiently with an appropriate concentration of glucose. The effectiveness of the vector was demonstrated by efficient screening of the amylase gene from a Bacillus subtilis genomic library. This vector system is expected to provide a more efficient and economic screening of bioactive products from DNA libraries in large quantities.

• Molecules and Cells
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• 제28권6호
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• pp.545-551
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• 2009

Mitogen-activated protein kinases (MAPK) affect the activation of activator protein-1 (AP-1), which plays an important role in regulating a range of cellular processes. However, the roles of these signaling factors on hydrogen peroxide ($H_2O_2$)-induced cell death are unclear. This study examined the effects of $H_2O_2$ on the activation of MAPK and AP-1 by exposing the cells to $H_2O_2$ generated by either glucose oxidase or a bolus addition. Exposing BJAB or Jurkat cells to $H_2O_2$ affected the activities of MAPK differently according to the method of $H_2O_2$ exposure. $H_2O_2$ increased the AP-1-DNA binding activity in these cells, where continuously generated $H_2O_2$ led to an increase in mainly the c-Fos, FosB and c-Jun proteins. The c-Jun-$NH_2$-terminal kinase (JNK)-mediated activation of c-Jun was shown to be related to the $H_2O_2$-induced cell death. However, the suppression of $H_2O_2$-induced oxidative stress by either JNK inhibitor or c-Jun specific antisense transfection was temporary in the cells exposed to glucose oxidase but not to a bolus $H_2O_2$. This was associated with the disruption of death signaling according to the severe and prolonged depletion of reduced glutathione. Overall, these results suggest that $H_2O_2$ may decide differently the mode of cell death by affecting the intracellular redox state of thiol-containing antioxidants, and this depends more closely on the duration exposed to $H_2O_2$ than the concentration of this agent.

• 한국발생생물학회지:발생과생식
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• 제27권4호
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• pp.185-193
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• 2023

Although increasing evidence of cause-and-effect relationship between BPA exposure and female reproductive disorders have been suggested through many studies, the precise biochemical and molecular mechanism(s) by which BPA interferes with steroidogenesis in the ovarian cells still remain unclear. Therefore, the purpose of this study was to discover the steroidogenic biomarker(s) associated with BPA treatment in human granulosa cell line, KGN. In this study, our results obtained via the analysis of steroidogenesis-related protein expression in KGN cells using quantitative polymerase chain reaction (qPCR) and western blot analyses revealed that the expression levels of steroidogenic acute regulatory (StAR) and aromatase decreased considerably and gradually after BPA treatment in a dose-dependent manner under BPA treatment. Further, remarkable decreases in their expression levels at the cellular levels were also confirmed via immunocytochemistry, and subsequent StAR and aromatase mRNA expression levels showed profiles similar to those observed for their proteins, i.e., both StAR and aromatase mRNA expression levels were significantly decreased under BPA treatment at concentrations ≥0.1 μM. We observed that follicle stimulating hormone upregulated StAR and aromatase protein expression levels; however, this effect was suppressed in the presence of BPA. Regarding the steroidogenic effects of BPA on KGN cells, controversies remain regarding the ultimate outcomes. Nevertheless, we believe that the results here presented imply that KGN cells have a good cellular and steroidogenic machinery for evaluating endocrine disruption. Therefore, StAR and aromatase could be stable and sensitive biomarkers in KGN cells for the cellular screening of the potential risk posed by exogenous and environmental chemicals to female reproductive (endocrine) function.

• 생명과학회지
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• 제26권12호
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• pp.1458-1465
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• 2016

본 연구는 oxidative stress에 의한 세포죽음 분석의 이상적인 모델로 사용되는 HT22세포를 이용하여 천문동 에탄올추출물의 glutamate에 의한 oxidative toxicity에 대한 신경보호 효과를 살펴보았다. 이를 위해 cell viability, lactate dehydrogenase (LDH), 그리고 세포죽음형태, reactive oxygen species (ROS), mitochondria membrane potential (MMP) 등에 대한 flow cytometry 및 Western blot분석을 이용하였다. 천문동 추출물의 처리는 cell viability 및 LDH분석에서 glutamate에 의한 cell toxicity를 저하시키며, 특히 apoptotic cell death를 현저히 감소시켰다. ROS 및 MMP분석 결과, 천문동 추출물은 ROS의 형성을 저하시키며 glutamate에 의해 저하된 MMP를 현저히 회복시켜 주었다. 이와 관련된 단백질 발현을 보면, 천문동 추출물은 PARP 및 HO-1의 발현을 억제하였다. 이상의 결과는 천문동 추출물이 HT22해마세포에서 ROS형성저해 및 MMP회복에 의해 세포죽음을 완화시켜 보호작용을 하는 것으로 사료되며 oxidative toxicity관련 질환에 적용 가능할 것으로 보여 진다.

• Journal of Chest Surgery
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• 제2권2호
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• pp.167-167
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• 1969

Two groups of esophagus graft were done in canine esophagus in 34 adult mongrel dogs. For the first group segmental replacement graft was done with fresh autologous pericardium tube, and for the second, patch graft was done utilizing fresh autologous pericardium, fresh homologous pericardium,and dacron piece. All eight dogs in the first segmental replacement graft group died 2 to 5 days after operation with severe empyema caused by anastomosis disruption. Among 26 patch graft dogs 2 died during operation and 7 died 13 to 18 days after operation. For the 17 long-term patch grafted survivors esophagography and postoperative weight check were done. Postoperative stool was collected and examined for dacron patch excretion. One, two, three, and four months postoperative long-term survivors were sacrificed to obtain specimens in each group respectively and the following observations were made.I. Survival; Autologous pericardium patch group showed no mortality but in homologous pericardium and dacron patch group only two thirds were long-term survivors. II. Postoperative swallowing; There was no case which demonstrated postoperative dysphagia. About half of the cases showed postoperative weight increase and in only 3 cases weight decrease followed operation. III. Dacron patch was excreted in the stool 8 to 23 days after operation. Animals which excreted dacron patch up to 9 days after operation all died of empyema due to anastomosis disruption.IV. Postoperative esophagogram; All esophagograms in each group showed no leakage of barium, no passage disturbances and no remarkable stenotic signs.V. Morphological findings; [A] Macroscopical findings; In one month group specimens of each group dense adhesion with surrounding structures was noted and luminal surface was smooth with contraction of the patched area. In two month groups anastomosis sutures were still exposed but patched area showed lesser abnormality. In three to four months groups sutures were covered completely and patched area showed only very slight signs of contraction. [B] Microscopic findings; In one month group luminal surface of the replaced tissue [transplanted tissue] showed almost complete epithelial covering that is composed of several layers of squamous cells with no evidence of keratinization. Basement membrane was also well distinct throughout. Slight to minimal inflammatory cells comprising of large mononuclears, lymphocytes and plasma cells were observed in the subepithelial fibrous stroma consisted entirely of loose fibrous tissue containing many newly formed capillaries and fibroblastic proliferation. Scattered suture granulomas were found, few of which became acutely inflamed. In two months group repairing process progressed with lesser degree of inflammatory cell infiltration and young capillary proliferation. Fibrous tissue was more matured showing even focal collagenization.Suture granuloma persisted but with lesser reactive changes. Epithelial covering was that of a mature non-keratinizing stratified squamous epithelium. In three and four months groups the replaced area showed essentially similar histological findings. However, subepithelial stroma still consisted entirely of connective tissue without evidence of smooth muscle regeneration. In this group, inflammatory cell infiltration was minimal or negligible. Among these patch materials autologous pericardium group showed the most satisfactory repairing process.The above mentioned results may signify the feasibility of autogenous pericardium patch graft in clinical esophageal surgery.

• Journal of Chest Surgery
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• 제2권2호
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• pp.168-186
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• 1969

Two groups of esophagus graft were done in canine esophagus in 34 adult mongrel dogs. For the first group segmental replacement graft was done with fresh autologous pericardium tube, and for the second, patch graft was done utilizing fresh autologous pericardium, fresh homologous pericardium,and dacron piece. All eight dogs in the first segmental replacement graft group died 2 to 5 days after operation with severe empyema caused by anastomosis disruption. Among 26 patch graft dogs 2 died during operation and 7 died 13 to 18 days after operation. For the 17 long-term patch grafted survivors esophagography and postoperative weight check were done. Postoperative stool was collected and examined for dacron patch excretion. One, two, three, and four months postoperative long-term survivors were sacrificed to obtain specimens in each group respectively and the following observations were made. I. Survival; Autologous pericardium patch group showed no mortality but in homologous pericardium and dacron patch group only two thirds were long-term survivors. II. Postoperative swallowing; There was no case which demonstrated postoperative dysphagia. About half of the cases showed postoperative weight increase and in only 3 cases weight decrease followed operation. III. Dacron patch was excreted in the stool 8 to 23 days after operation. Animals which excreted dacron patch up to 9 days after operation all died of empyema due to anastomosis disruption. IV. Postoperative esophagogram; All esophagograms in each group showed no leakage of barium, no passage disturbances and no remarkable stenotic signs. V. Morphological findings; [A] Macroscopical findings; In one month group specimens of each group dense adhesion with surrounding structures was noted and luminal surface was smooth with contraction of the patched area. In two month groups anastomosis sutures were still exposed but patched area showed lesser abnormality. In three to four months groups sutures were covered completely and patched area showed only very slight signs of contraction. [B] Microscopic findings; In one month group luminal surface of the replaced tissue [transplanted tissue] showed almost complete epithelial covering that is composed of several layers of squamous cells with no evidence of keratinization. Basement membrane was also well distinct throughout. Slight to minimal inflammatory cells comprising of large mononuclears, lymphocytes and plasma cells were observed in the subepithelial fibrous stroma consisted entirely of loose fibrous tissue containing many newly formed capillaries and fibroblastic proliferation. Scattered suture granulomas were found, few of which became acutely inflamed. In two months group repairing process progressed with lesser degree of inflammatory cell infiltration and young capillary proliferation. Fibrous tissue was more matured showing even focal collagenization. Suture granuloma persisted but with lesser reactive changes. Epithelial covering was that of a mature non-keratinizing stratified squamous epithelium. In three and four months groups the replaced area showed essentially similar histological findings. However, subepithelial stroma still consisted entirely of connective tissue without evidence of smooth muscle regeneration. In this group, inflammatory cell infiltration was minimal or negligible. Among these patch materials autologous pericardium group showed the most satisfactory repairing process. The above mentioned results may signify the feasibility of autogenous pericardium patch graft in clinical esophageal surgery.