• Biomolecules & Therapeutics
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• 제22권6호
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• pp.510-518
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• 2014

Chronic (>24 h) exposure of arsenite, an environmental toxicant, has shown the decreased nitric oxide (NO) production in endothelial cells (EC) by decreasing endothelial NO synthase (eNOS) expression and/or its phosphorylation at serine 1179 ($eNOS-Ser^{1179}$ in bovine sequence), which is associated with increased risk of vascular diseases. Here, we investigated the acute (<24 h) effect of arsenite on NO production using bovine aortic EC (BAEC). Arsenite acutely increased the phosphorylation of $eNOS-Thr^{497}$, but not of $eNOS-Ser^{116}$ or $eNOS-Ser^{1179}$, which was accompanied by decreased NO production. The level of eNOS expression was unaltered under this condition. Treatment with arsenite also induced reactive oxygen species (ROS) production, and pretreatment with a ROS scavenger N-acetyl-L-cysteine (NAC) completely reversed the observed effect of arsenite on $eNOS-Thr^{497}$ phosphorylation. Although protein kinase C (PKC) and protein phosphatase 1 (PP1) were reported to be involved in $eNOS-Thr^{497}$ phosphorylation, treatment with PKC inhibitor, Ro318425, and overexpression of various PKC isoforms did not affect the arsenite-stimulated $eNOS-Thr^{497}$ phosphorylation. In contrast, treatment with PP1 inhibitor, calyculin A, mimicked the observed effect of arsenite on $eNOS-Thr^{497}$ phosphorylation. Lastly, we found decreased cellular PP1 activity in arsenite-treated cells, which was reversed by NAC. Overall, our study demonstrates firstly that arsenite acutely decreases NO production at least in part by increasing $eNOS-Thr^{497}$ phosphorylation via ROS-PP1 signaling pathway, which provide the molecular mechanism underlying arsenite-induced increase in vascular disease.

• Journal of Ginseng Research
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• 제47권1호
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• pp.123-132
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• 2023

Background: Pseudotyped virus systems that incorporate viral proteins have been widely employed for the rapid determination of the effectiveness and neutralizing activity of drug and vaccine candidates in biosafety level 2 facilities. We report an efficient method for producing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pseudovirus with dual luciferase and fluorescent protein reporters. Moreover, using the established method, we also aimed to investigate whether Korean Red Ginseng (KRG), a valuable Korean herbal medicine, can attenuate infectivity of the pseudotyped virus. Methods: A pseudovirus of SARS-CoV-2 (SARS-2pv) was constructed and efficiently produced using lentivirus vector systems available in the public domain by the introduction of critical mutations in the cytoplasmic tail of the spike protein. KRG extract was dose-dependently treated to Calu-3 cells during SARS2-pv treatment to evaluate the protective activity against SARS-CoV-2. Results: The use of Calu-3 cells or the expression of angiotensin-converting enzyme 2 (ACE2) in HEK293T cells enabled SARS-2pv infection of host cells. Coexpression of transmembrane protease serine subtype 2 (TMPRSS2), which is the activator of spike protein, with ACE2 dramatically elevated luciferase activity, confirming the importance of the TMPRSS2-mediated pathway during SARS-CoV-2 entry. Our pseudovirus assay also revealed that KRG elicited resistance to SARS-CoV-2 infection in lung cells, suggesting its beneficial health effect. Conclusion: The method demonstrated the production of SARS-2pv for the analysis of vaccine or drug candidates. When KRG was assessed by the method, it protected host cells from coronavirus infection. Further studies will be followed for demonstrating this potential benefit.

• 시큐리티연구
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• 제22호
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• pp.113-144
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• 2010

생물공학과 첨단과학의 발전으로 생물무기의 파괴력이 증대되고 활용이 용이해짐에 따라 생물무기가 테러집단에게 매력적인 공격수단이 되고 있는 가운데 우리나라에서는 국내외 테러위협이 고조되고 있는데다 북한의 대남 생물테러 가능성도 잠재하고 있어 효과적인 대응체제 마련이 긴요한 실정이다. 이에 따라 대응체제 제시에 필요한 시사점을 얻기 위해 생물테러 위협에 가장 잘 대비하고 있는 것으로 평가받고 있는 미국의 체제를 분석하여 (1) 생물테러 관계 법률을 제정하거나 현실에 맞게 정비하고 있으며 (2) 국토안보부 중심으로 대테러업무를 통합하여 대응역량 극대화를 추구하고 있고 (3) 생물테러 대응을 위한 절차와 요령 등을 매뉴얼화하여 실무대응능력을 제고하고 있다는 특징을 발견하였다. 이어서 우리나라의 대응체계를 법령체제, 역할분담, 협조관계, 자원운용 부분으로 나누어 실태를 살펴본 결과, 법제도가 허술하고 통합대응기구가 부재한데다 대응시설과 물자부족 및 인력관리가 미흡하다는 문제점을 도출하였다. 이와 같은 미국제도가 주는 시사점과 우리나라 대응체제의 문제점을 토대로 (1) 생물테러 기본법이 될 테러방지법의 제정과 생물테러대응과 관계되는 재난대비 법령 등의 개정이 필요하며 (2) 정부 각 부처에 분산되어 있는 대테러 업무가 기능적으로 하나로 모아져 대응역량을 최고화 할 수 있게 대테러 관계부처를 강력히 조정할 권한을 가진 통합대테러조직을 구축하고 (3) 생물안전 수준이 높은 실험실 설치 및 전략적 비축의약품을 충분히 확보하며 (4) 대응에 필요한 세부적인 내용을 담아 매뉴얼을 작성 활용함으로써 생물테러에 경험이 없는 우리 대응요원의 한계를 보완하는 등의 개선방안을 제시하였다.