• Biomolecules & Therapeutics
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• v.22 no.6
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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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• v.47 no.1
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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.

• Korean Security Journal
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• no.22
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• pp.113-144
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• 2010

Bioterrorism is becoming more attractive to terrorist groups owing to the dramatic increase in the utility and lethality of biological weapons in line with today's cutting-edge biological science and technology. The Republic of Korea is facing both internal and external terrorist threats, as well as the possible biological warfare by North Korea. Therefore, it is essential to establish an effective bioterrorism response system in the ROK. In order to come up with the adequate response system for the ROK, an in-depth study has been conducted on the current bioterrorism response system of the U.S. whose preparedness is considered relatively adamant. As a result, the following facts have been found: (1)the legislation with regard to bioterrorism has been established or amended according to the current situation in the U.S., (2)the counter terrorism activities have been integrated with the Department of the Homeland Security as the central agency in order to maximize the national CT capacity, (3)Specific procedures and instructions to cope with bioterrorism have been made into manuals so as to enhance the working-level response capabilities. Next, the analysis on the ROK's bioterrorism response system has been performed in various categories, including the legislation system, task role distribution, cooperative relations, and resource application. It turned out that the ROK's legislation basis is relatively weak and it lacks the apparatus to integrate the bioterrorism response activities on the national level. The shortage of the adequate response facilities and resources, as well as the poor management of manpower have also emerged as problems that hinder the effective CT implementations. Through an analytical and comparative study of the U.S. and the ROK systems, this paper presents several ways to ameliorate improve the current system in the ROK as follows: (1)establish the anti-terrorism law, which would be the basic legal basis for the bioterrorism-related matters; and make revisions to the disaster-related legislation, relevant to bioterrorism response activities, (2)establish an integrated body that has a powerful authority to coordinate the relevant CT agencies; and converge the decentralized functions to maximize the overall response capacity, (3)install the laboratories with a high biosafety level and secure enough of the strategic medical stock-pile, (4)enhance the ability of the inexperienced response personnel by providing with a manual that has detailed instructions.