• Herbal Formula Science
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• v.26 no.3
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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.

• The Journal of the Korean Society for Microbiology
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• v.11 no.1
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• pp.1-11
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• 1976

An electron microscopic study has been made of the effects of change of cell structure of Escherichia coli treated with several disinfectants. The alterations observed as follows: 1. Nucleoid, cytoplasm which contain ribosomes and cell wall appeared to be composed of a parallel triple. layered membrane can observed in control Escherichia coli. 2. The nuclear material was no longer demonstrable in its normal sites. The cytoplasm lost its granularity, became homogeneous and disruption of cell wall were observed by the treatment with 70% ethyl alcohol and 3% $H_2O_2$. 3. Aggregation of ribosomes and condensation of nuclear material were also observed by the treatment of 5% lysol solution and 1% dodecyl-diamino-ethyl-glycin-hydrochloride. 4. In autoclaved group, the each layer of cell wall was separated and destroyed in some sites where cytoplasm was extracted.

• Journal of Microbiology and Biotechnology
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• v.17 no.7
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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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• v.28 no.6
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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.

• Development and Reproduction
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• v.27 no.4
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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.

• Journal of Life Science
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• v.26 no.12
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• pp.1458-1465
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• 2016

We investigated the neuroprotective effect of an ethanol extract from Asparagus cochinchinensis (AC) against glutamate-induced toxicity in the HT22 hippocampal cell, which is an ideal in vitro model for oxidative stress. The neuroprotective effects of AC in HT22 cells were evaluated by analyzing cell viability, lactate dehydrogenase (LDH), flow cytometry for cell death types, reactive oxygen species (ROS), mitochondria membrane potential (MMP), and Western blot assays. In the cell death analysis, AC treatment resulted in significantly attenuated glutamate-induced loss of cell viability with a decrease in LDH release. AC treatment also reduced glutamate-induced apoptotic cell death. In the ROS and MMP analysis, AC treatment inhibited the elevation of intracellular ROS induced by glutamate exposure and the disruption of MMP. In oxidative stress-related proteins analysis, AC treatment inhibited the expression of poly ADP ribose polymerase and heme oxygenase-1 by glutamate. These results indicate that AC exerts a significant neuroprotective effect against glutamate-induced hippocampal damage by decreasing ROS production and stabilizing MMP. Thus, AC potentially provides a new strategy for the treatment of oxidative stress-related diseases.

• Journal of Chest Surgery
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• v.2 no.2
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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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• v.2 no.2
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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.

• KSBB Journal
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• v.30 no.6
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• pp.291-295
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• 2015

The limited productivity of natural shell matrix proteins has hampered the investigation of their biochemical properties and practical applications, although biominerals in nature obtained by organic-inorganic assemblies have attractive mechanical and biological properties. Here, we prepared a vector for the expression of a fusion protein of a shell matrix protein from Pinctada fucata (named as GRP_BA) with the GRGDSP residue. The fusion protein of BA-RGD was simply produced in E. coli and purified through sequential steps including the treatment with $CaCl_2$ and EDTA solution for cell membrane washing, mechanical cell disruption and the application of non-ionic surfactant of Triton X-100 for BA-RGD inclusion body washing. The production yield was approximately 60 mg/L, any other protein band was not observed in SDS-PAGE and it was estimated that above 97% endotoxin was removed compared to the endotoxin level of whole cell. This study showed this simple and easy purification approach could be applied to the purification of BA-RGD fusion protein. It is expected that the protein could be utilized for the preparation of biominerals in practical aspects.

• The Journal of the Korean bone and joint tumor society
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• v.10 no.1
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• pp.29-33
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• 2004

A giant cell tumor (GCT) of the distal radius is not common. Curettage with bone cementation is considered as a treatment of choice but, in the case of recurrence, marked cortical disruption, or articular invasion, en bloc excision and reconstruction with proximal fibular bone graft is usual procedure. In reconstruction of en bloc resected distal radius which had recurred GCT after conservative operation, we used the ultrahigh molecular weight polyethylene (UHMWPE) liner with intramedullary rod and bone cement, because the contamination was extent in previous operation and recurrence after fibular bone graft was fearful. This article introduce our new surgical procedure.