• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2000년도 International Meeting 2000
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• pp.23-36
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• 2000

Japanese encephalitis virus (JEV) is the causative agent of a mosquito-borne encephalitis and is transmitted to human via persistently infected mosquito vectors. Although the virus is known to cause only acute infection, there were reports that showed neurological sequelae, latent infection in peripheral mononuclear cells, and recurrence of the disease after acute encephalitis. Innate resistance of certain cell lines, abnormal SN1 expression of the virus, and anti-apoptotic effect of cullular bcl-2 have been suggested as probable causes of JEV persistence even in the absence of defective interfering (DI) particles. Although possible involvement of DI particles in JEV persistence was suggested, neither has a direct evidence for DI presence nor its molecular characterization been made. Two questions asked in this study are whether the DI virus plays any role in JEV persistent infection if it is associated with and what type of change(s) can be made in persistently infected cells to avoid apoptosis even with the continuous virus replication, DI-free standard stock of JEV was infected in BHK-21, Vero, and SW13 cells and serial high multiplicity passages were performed in order to generate DI particles. There different-sized DI RNA species which were defective in both structural and nonstructural protein coding genes. Rescued ORFs of the DI genome maintained in-frame and the presence of replicative intermediate or replicative form RNA of the DI particles confirmed their replication competence. On the other hand, several clones with JEV persistent infection were established from the cells survived acute infections during the passages. Timing of the DI virus generation during the passages seemed coincide to the appearance of persistently infected cells. The DI RNAs were identified in most of persistently infected cells and were observed throughout the cell maintenance. One of the cloned cell line maintained the viral persistence without DI RNA coreplication. The cells with viral persistence released the reduced but continuous infectious JEV particle for up to 9 months and were refractory to homologous virus superinfection but not to heterologous challenges. Unlike the cells with acute infection these cells were devoid of characteristic DNA fragmentation and JEV-induced apoptosis with or without homologous superinfection. Therefore, the DI RNA generated during JEV undiluted serial passage on mammalian cells was shown to be biologically active and it seemed to be responsible, at least in part, for the establishment and maintenance of the JEV persistence in mammalian cells. Viral persistence without DI RNA coreplication, as in one of the cell clones, supports that JEV persistent infection could be maintained with or without the presence of DI particles. In addition, the fact that the cells with JEV persistence were resistant against homologous virus superinfection, but not against heterologous one, suggests that different viruses have their own and independent pathway for cytopathogenesis even if viral cytopathic effect could be converged to an apoptosis after all.

• 대한한의학회지
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• 제26권1호
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• pp.174-186
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• 2005

Objectives : The water extract of Samul-tang (SMT) has traditionally been used for treatment of ischemic heart and brain damage in oriental medicine. However, little is known about the mechanism by which the water extract of SMT rescues cells from these damages. Methods: This study was designed to investigate the protective mechanisms of SMT on oxidative stress-induced toxicity in H9c2 cardiomyoblast cells. Treatment with $H_2O_2$ markedly induced death of H9c2 cardiomyoblast cells in a dose-dependent manner. Results: The characteristics of H20z-induced death of H9c2 showed apparent apoptotic features such as DNA fragmentation and morphological change. However, SMT significantly reduced both H202-induced cell death and morphological change. The decrease of Bc-2 expression by High were inhibited by SMT. In addition, the increase of Bax expression was also inhibited by SMT. The cotreatment of SMT and $H_2O_2$ in H9c2 cells also induced the phosphorylation of ERK in a time-dependent manner. Moreover, PD98059, a specific inhibitor of ERK1/2 attenuated the protective effects of SMT on $H_2O_2-induced$ toxicity in H9c2 cardiomyoblast cells. These results suggest that both ERK1/2 signaling pathways play important roles in the protective effects of SMT on $H_2O_2-induced$ apoptotic death of H9c2 cells. Also, the expression profile of proteins in $H_2O_2$ cardiomyoblast cells were screened by using two-dimensional (2-D) gel electrophoresis. Among 300 spots resolved in 2-D gels, the comparison of control versus apoptosis cells revealed that signal intensity of 17 spots increased and 11 spots decreased. Conclusions: Taken together, this study suggests that the protectiw effects of the water extract of SMT against oxidative damages may be mediated by the modulation of Bc1-2 and Bax expression via the regulation of the ERK signaling pathway.

• 고려인삼학회:학술대회논문집
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• 고려인삼학회 1998년도 Advances in Ginseng Research - Proceedings of the 7th International Symposium on Ginseng -
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• pp.231-243
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• 1998

In the present study, we provide evidence that ginsenoside $Rh_2$ (G-$Rh_2$) as well as staurosporine induces apoptosis of human hepatoma SK-HEP-1 cells by caspase 3-mediated processing of $p21^{WAFI/CIPI}$ in the early stage of apoptosls. Immunoblottings showed that G-$Rh_2$ as well as statrosporine induced the processing of caspase-3 to an active form, pl7. In stable Bcl-2 transfectants however, G-$Rh_2$ induced DNA fragmentation, while staurosporine did not. In the early stage of apoptosis, $p21^{WAFI/CIPI}$ was detected to undergo proteolytic processing specifically conducted by caspase-3. $p21^{WAFI/CIPI}$ translated in vitro was cleaved into a p14 fragment, when incubated with cell extracts obtained from either G-$Rh_2$- or staurosporine-treated cells. Cleavage was equally inhibited in both cases by adding Ac-DEVD-cho, a specific caspase-3 inhibitor, but not by Ac-YVkD-cho, a specific caspase-l inhibitor. Similarly, $p21^{WAFI/CIPI}$ was efficiently leaved by recombinant caspase-3 overexpressed in E. coli. Moreover, the endogenous $p21^{WAFI/CIPI}$ of untreated-cell extracts was also cleaved by recombinant caspase-3. Mutation analysis allowed identification of two caspase-3 cleavage sites, $DHVD^{112}$/L and $SMTD^{149}$/F, which are located within, or near the interaction domains for cyclins, Cdks, and PCNA. Taken together, these results show that G-$Rh_2$ as well as staurosporine increases caspase-3 activity, which in turn directly cleaves $p21^{WAFI/CIPI}$ resulting in elevation of Cdk kinase activity in the early stages of apoptosis. We propose that proteolytic cleavage of $p21^{WAFI/CIPI}$ is a functionally relevant event that allows unleashing the cyclin/Cdk activity from the inhibitor seen in the earlier stage of apoptosis, the event of which may be associated with the triggering mechanism for the execution of apoptosis.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 춘계학술발표회
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• pp.14-14
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• 2018

This Camptotheca acuminata Decne. (CA), belonging to Nyssaceae, is a deciduous tree. and has been used as Traditional Chinese medicine since ancient times. The CA produces camptothecin a natural indole alkaloid, and reported to have anti-cancer effects. But the studies on biological activities of CA leaves are insufficient. Therefore, this study confirmed various biological activities such as antioxidant, antidiabetic, anticancer, antiinflammatory and metabolism analysis by HPLC-MS/MS of CA leaves. The $RC_{50}$ values of DPPH radical scavenging activity of ethyl acetate fraction, n-Butanol fraction, methanol extraction, water fraction and n-Hexane fraction were $12.23{\pm}0.01$, $15.93{\pm}0.42$, $55.12{\pm}0.45$, $56.29{\pm}4.15$ and $427.29{\pm}6.13ug/mL$, respectively. The $IC_{50}$ values of ${\alpha}$-glucosidase inhibitory activity of ethyl acetate fraction, n-Butanol fraction, methanol extraction, n-Hexane fraction and water fraction were $24.29{\pm}0.14$, $47.86{\pm}0.45$, $54.23{\pm}1.21$ $466.76{\pm}2.21$ and $623.91{\pm}9.67ug/mL$, respectively. The nitric oxide release activity of n-Hexane fraction, methanol extraction, ethyl acetate fraction, water fraction and n-Butanol fraction were $31.49{\pm}5.74$, $29.79{\pm}0.71$, $26.89{\pm}0.71$, $8.24{\pm}5.83$ and $7.75{\pm}4.08%$ at 25 ug/mL, respectively. The anti-cancer activity of n-Hexane fraction, methanol extraction, ethyl acetate fraction, water fraction and n-Butanol fraction were $31.49{\pm}5.74$, $29.79{\pm}0.71$, $26.89{\pm}0.71$, $8.24{\pm}5.83$ and $7.75{\pm}4.08%$ at 25 ug/mL, respectively. The ethyl acetate fraction activities showed higher biological activities than other fractions. Thus, Additional studies were conducted using ethyl acetate fraction. Metabolite analysis was performed using a LCMS-8040 triple quadrupole mass spectrometer. As a result, Five compounds (1-5) were identified in the ethyl acetate fraction of the CA leave. The identification of these compounds was generated by the analysis of fragmentation methods of the negative and positive ion modes. Five compounds were identified as gallic acid (1), chlorogenic acid (2), isoquercetin (3), astragalin (4) and camptothecin (5). These results suggest that the CA leave can be used for functional materials.

• 한국지리정보학회지
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• 제16권2호
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• pp.47-64
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• 2013

본 연구에서는 위성 자료를 기반으로 생성된 토지피복 정보를 시계열적인 분석을 통해 지난 30년간 부산 경남 지역의 경관 구조 변화를 추적하고 해당 지역 기온 변화와의 상관성을 파악하였다. 토지 피복 자료의 기하학적 구조를 정량화하기 위해 경관구조 분석 프로그램인 FRAGSTATS를 사용하여 토지피복별 경관지수를 산출하였다. 이를 통해 주요 토지이용의 변화상과 개발에 따른 경관의 분절화 과정을 확인하였다. 부산과 경남 지역의 10개 측후소 자료에 따르면, 부산의 평균 기온은 1980년대 $14.1^{\circ}C$에서 2000년대 $14.8^{\circ}C$로 증가하였고, 경남 지역의 평균 기온은 1980년대 $13.2^{\circ}C$에서 2000년대 $13.9^{\circ}C$로 증가하였다. 이러한 장기 기온 변화패턴은 도시화와 같은 우리나라 동남부의 전형적인 경관구조 변화와 상관성을 갖는 것으로 판단된다. 부산시 분석 자료에 의하면, 지난 30년간 도시역은 전체 면적의 9.7%에서 26.8%로 증가하였고, 산림과 농경지는 각각 58%에서 48.4%로, 28.3%에서 13.4%로 감소하였다. 해당 기간별 기온자료와 비교해보면, 도시역이 증가하고 산림과 농경지 면적이 꾸준히 감소함에 따라 부산의 평균기온이 1990년대 이후 큰 폭으로 증가하였다. 경남지역의 도시역 면적은 12배 이상 증가해 매우 빠른 도시화를 겪었다. 도시화에 따른 기온 변화는 지역적으로 구분되었는데, 경남 해안지역은 기온상승의 폭이 가장 낮았고 중부경남에 이어 내륙 산악지역에서 가장 높은 기온증가율이 나타났다.

• International Journal of Oral Biology
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• 제34권1호
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• pp.7-14
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• 2009

Nitric oxide (NO) acts as an intracellular messenger at the physiological level but can be cytotoxic at high concentrations. The cells within periodontal tissues, such as gingival and periodontal fibroblasts, contain nitric oxide syntheses and produce high concentrations of NO when exposed to bacterial lipopolysaccharides and cytokines. However, the cellular mechanisms underlying NO-induced cytotoxicity in periodontal tissues are unclear at present. In our current study, we examined the NO-induced cytotoxic mechanisms in human gingival fibroblasts (HGF). Cell viability and the levels of reactive oxygen species (ROS) were determined using a MTT assay and a fluorescent spectrometer, respectively. The morphological changes in the cells were examined by Diff-Quick staining. Expression of the Bcl-2 family and Fas was determined by RT-PCR or western blotting. The activity of caspase-3, -8 and -9 was assessed using a spectrophotometer. Sodium nitroprusside (SNP), a NO donor, decreased the cell viability of the HGF cells in a dose- and time-dependent manner. SNP enhanced the production of ROS, which was ameliorated by NAC, a free radical scavenger. ODQ, a soluble guanylate cyclase inhibitor, did not block the SNP-induced decrease in cell viability. SNP also caused apoptotic morphological changes, including cell shrinkage, chromatin condensation, and DNA fragmentation. The expression of Bax, a member of the proapoptotic Bcl-2 family, was upregulated in the SNP-treated HGF cells, whereas the expression of Bcl-2, a member of the anti-apoptotic Bcl-2 family, was downregulated. SNP augmented the release of cytochrome c from the mitochondria into the cytosol and enhanced the activity of caspase-8, -9, and -3. SNP also upregulated Fas, a component of the death receptor assembly. These results suggest that NO induces apoptosis in human gingival fibroblast via ROS and the Bcl-2 family through both mitochondrial- and death receptor-mediated pathways. Our data also indicate that the cyclic GMP pathway is not involved in NO-induced apoptosis.

• 수면정신생리
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• 제4권2호
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• pp.191-200
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• 1997

Objectives : Periodic limb movements during sleep (PLMS) may cause arousals that may lead to non-restorative sleep. PLMS is characterized by long sleep latency, sleep fragmentation, frequent stage shifts, and rarity of stages 3/4 NREM sleep on polysomnography. However, controversies have existed and it still remains to be elaborated whether PLMS actually causes insomnia, since normal persons happen to have PLMS. Clinically, it would be crucial to know factors which might disturb sleep in PLMS. We became interested in Coleman's theory(1980) that invariant periodic movements disturb patients' sleep less. Though, Coleman's study seems to have been confounded by including PLMS patients with various co-morbid sleep disorders. Therefore, we attempted to study in patients only with PLMS the effects of movement patterns on sleep architecture. Methods : In 27 patients diagnosed as having PLMS only with clinical interview and nocturnal polysomnography, we studied the relationship between the movement patterns such as mean duration and variability of periodic limb movement's interval and the sleep architecture variables. Results : The shorter and the more regular the limb movement intervals were, the fewer arousals followed. The movement intervals of the older patients were shorter and more regular than the younger patients. The probability of the accompanying arousal with each limb movement increased as the duration and variance of the movement intervals increased. It decreased as the age and the frequency of limb movements increased. Among these factors the most significant one was the mean duration of the movement intervals. In other words, the shorter the movement intervals were, the less disturbed sleep was. Conclusion : PLMS frequency increases with aging but the probability of the accompanying arousal with each movement decreases with aging. Sleep-disturbing effects of PLMS depends more on the duration and variability of movement intervals than the PLMS frequency.

• 한국식품영양과학회지
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• 제30권6호
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• pp.1152-1157
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• 2001

도축 후 24시간 냉장한 상등급(B2 등급)과 등외등급(D 등급) 쇠고기 지육에서 등심부위를 분할하여 두 쪽으로 나누어 한 쪽은 랩으로 싸서 1일간 냉장하고, 다른 한 쪽은 500g씩 자르고 진공포장하여 30일간 냉장하여 전자는 신선육, 후자는 냉장육이라 하였다. 상등급육과등외등급육의 기호특성을 비교하고 등외등급육의 냉장효과를 알아보기 위하여 신선육과 냉장육의 색도, 연도, ATP 관련물질, 지방산 조성 및 관능평가에 대한 실험을 하였다. 신선육의 경우 상등급육이 등외등급육에 비하여 pH. 젖산함량, 미오글로빈 함량이 낮고 표면색이 우수하였다. 상등급육은 경도와 저작성이 낮으면서 근원섬유 소편화도가 높아 연한 현상을 보였다. 상등급육은 포화지방산 함량이 낮고 단일 불포화지방산에 대한 포화지방산의 비율(MUFA/SFA)과 ATP 함량이 높고, 생육향이 우수하였다. 상등급육은 가열감량 및 가열단축도가 낮은 반면 가열육의 기호성이 현저히 우수하였다. 등외등급육은 상등급육에 비해서 경도, 저작성, 근원섬유 소편화도 및 MUFA/SFA에 대한 냉장효과가 크게 나타났다. 그러나 냉장육의 연도, 가열육 향 그리고 기호성이 등외등급육에 비해 상등급육이 현저히 우수하였다.

• 한국식품영양과학회지
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• 제16권3호
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• pp.55-61
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• 1987

산란노계육(産卵老鷄肉)(Arbor Acres, 72주령(週齡))의 가슴근육과 다리근육을 냉장 및 동결저장하면서 pH, 단백질의 추출성, 근원섬유단백질의 ATPase 활성, 소편화, 냉동감량 및 드립량의 변화를 비교하였다. 가슴근육과 다리근육의 pH는 각각 냉장 1일 및 동결저장 1주에 가장 낮았다. 근원섬유단백질의 추출성은 냉장기간이 경과하면서 점차 증가하였으며, 동결저장에서는 1주에 가장 높은 수준을 보였다. 근원섬유단백질의 $Mg^{2+}-ATPase$활성은 냉장 1일 및 동결저장 1주에 각각 높게 나타났다. 근원섬유의 소편화 정도는 냉장 1일 및 동결저장 1주에 크게 변화하였으며 냉동감량과 드립량은 동결저장 기간이 경과하면서 점차 많아졌다. 냉장 및 동결저장 중 가슴근육은 다리근육에 비하여 pH가 낮았고, 근원섬유단백질의 추출성, $Mg^{2+}-ATPase$활성, 드립량 및 소편화 정도가 높았으나 저장기간 중 변화되는 양상은 비슷하였다.

• BMB Reports
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• 제37권2호
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• pp.185-191
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• 2004

Tissue transglutaminase (tTGase) regulates various biological processes, including extracellular matrix organization, cellular differentiation, and apoptosis. Here we report the protective role of tTGase in the cell death that is induced by the tumor necrosis factor $\alpha$ (TNF-$\alpha$) and ceramide, a product of the TNF-$\alpha$ signaling pathway, in human neuroblastoma SH-SY5Y cells. Treatment with retinoic acid (RA) induced the differentiation of the neuroblastoma cells with the formation of extended neurites. Immunostaining and Western blot analysis showed the tTGase expression by RA treatment. TNF-$\alpha$ or $C_2$ ceramide, a cell permeable ceramide analog, induced cell death in normal cells, but cell death was largely inhibited by the RA treatment. The inhibition of tTGase by the tTGase inhibitors, monodansylcadaverine and cystamine, eliminated the protective role of RA-treatment in the cell death that is caused by TNF-$\alpha$ or $C_2$-ceramide. In addition, the co-treatment of TNF-$\alpha$ and cycloheximide ecreased the protein level of tTGase and cell viability in the RA-treated cells, supporting the role of tTGase in the protection of cell death. DNA fragmentation was also induced by the co-treatment of TNF-$\alpha$ and cycloheximide. These results suggest that tTGase expressed by RA treatment plays an important role in the protection of cell death caused by TNF-$\alpha$ and ceramide.