Aggravation of post-ischemic liver injury by overexpression of A20, an NF-κB suppressor

Yu, J.,Lee, H.S.,Lee, S.M.,Yu, H.C.,Moon, W.S.,Chung, M.J.,Park, J.W.,Park, B.H. Elsevier Science Publishers 2011 Journal of hepatology Vol.55 No.2

Backgroud & Aims: A20 is an intracellular ubiquitin-editing enzyme that plays an important role in the negative feedback regulation of NF-κB activation in response to a diverse range of stimuli. Liver ischemia/reperfusion injury is associated with rapid activation of NF-κB signaling, but the role of NF-κB in hepatic ischemia/reperfusion injury remains controversial. The NF-κB signaling pathway mediates both protective and deleterious effects in the liver. Here, we examined whether A20 inhibited or aggravated hepatic ischemia/reperfusion injury. Methods: We used IκBα super-repressor as a positive control and overexpressed A20 and IκBα super-repressor in the liver of C57BL/6 mice. Mice underwent 45min of partial hepatic ischemia and were then reperfused. Results: Protein level of A20 was increased after reperfusion. Mice subjected to ischemia/reperfusion injury showed increased NF-κB activation, as evidenced by phosphorylation of IκBα and nuclear translocation of NF-κB. Prior transfection with Ad-A20 or Ad-IκBα super-repressor attenuated NF-κB activation and aggravated liver injury. Serum aminotransferases and proinflammatory cytokines, hepatocellular necrosis, and hepatic neutrophil infiltration were markedly increased compared to those of uninfected or control virus infected mice. In addition, A20 abolished the beneficial effect of ischemic preconditioning. Conclusions: Our results suggest that inhibition of NF-κB activation by A20 aggravated partial hepatic ischemia/reperfusion injury. Understanding how the NF-κB pathway plays a role in directing a clinical outcome may lead to better prospects of more rational approaches to reduce post-ischemic liver injury.

Purification and characterization of a novel fibrinolytic α chymotrypsin like serine metalloprotease from the edible mushroom, Lyophyllum shimeji

Moon, S.M.,Kim, J.S.,Kim, H.J.,Choi, M.S.,Park, B.R.,Kim, S.G.,Ahn, H.,Chun, H.S.,Shin, Y.K.,Kim, J.J.,Kim, D.K.,Lee, S.Y.,Seo, Y.W.,Kim, Y.H.,Kim, C.S. Society for Bioscience and Bioengineering, Japan ; 2014 Journal of bioscience and bioengineering Vol.117 No.5

A novel fibrinolytic enzyme was purified from Lyophyllum shimeji, a popular edible mushroom in Asia. The enzyme was purified using combination of anion exchange chromatography on a Mono Q 5/5 column and size exclusion gel filtration chromatography on Superdex 200 100/300 column. This purification protocol resulted 80.9-fold purification of the enzyme and a final yield of 5.7%. The molecular weight of the purified enzyme was estimated to be 21 kDa by SDS-PAGE and size exclusion gel filtration. The N-terminal amino acid sequence was found to be ITFQSASP, which is dissimilar from that of known fibrinolytic enzymes. The purified enzyme was a neutral protease with an optimal reaction pH and temperature of 8.0 and 37<SUP>o</SUP>C, respectively. Enzymatic activity was inhibited by Cu<SUP>2+</SUP> and Co<SUP>2+</SUP>. It was also significantly inhibited by PMSF and TPCK. Furthermore, it was found to exhibit a higher specificity for S-7388, a well-known chymotrypsin chromogenic substrate, indicating chymotrypsin like serine metalloprotease. The relative fibrinolytic activity of 5 μg purified enzyme have two fold more activity than 1 unit/ml of plasmin on fibrin plate. Furthermore, purified enzyme preferentially hydrolyzed the Aα-chain followed by the Bβ- and γ-chain of fibrinogen, which is precursor of fibrin. Therefore, these data suggests that the fibrinolytic enzyme derived from edible mushroom, L. shimeji, might be useful for thrombolytic therapy and preventing thrombotic disease.

심혈관질환의 위험인자로서의 endothelial nitric oxide synthase의 SNP 연구

윤수인,문제성,신철,박현영,이정복,조인호 국립보건연구원 2000 국립보건원보 Vol.37 No.-

Achaete-scute complex homologue 2 accelerates the development of Sjogren's syndrome-like disease in the NOD/ShiLtJ mouse

Kim, S.M.,Kwon, J.E.,Park, J.S.,Seo, H.B.,Jung, K.A.,Moon, Y.M.,Lee, J.,Kwok, S.K.,Cho, M.L.,Park, S.H. Elsevier/North-Holland Biomedical Press 2017 Immunology letters Vol.190 No.-

Achaete-scute complex homologue 2 (Ascl2) has been reported to induce the differentiation and activation of follicular helper T (T<SUB>FH</SUB>) cells, which are essential for development of Sjogren's syndrome (SS). This study examined whether Ascl2 plays a role in the development of SS. NOD/ShiLtJ mice were injected with an Ascl2-overexpression vector, and the infiltration of lymphocytes into salivary and lacrimal glands was assessed. The expression of inflammatory cytokines and chemoattractants for T or B cells was measured. The activation of T<SUB>FH</SUB> cells was assessed using a specific marker of T<SUB>FH</SUB> cells. Ascl2 level was also measured in SS patients. Overexpression of Ascl2 increased the expression of C-X-C chemokine receptor type 5 (CXCR5) in both salivary and lacrimal glands (p<0.0001). Overexpression of Ascl2 also increased the expression of proinflammatory cytokines and chemoattractants including interleukin 6 (IL-6), tumor necrosis factor-α, IL-8, programmed cell death 1 (PD-1), IL-21, and B-cell lymphoma 6 (Bcl-6). Overexpression of Ascl2 increased the populations of CD4<SUP>+</SUP>CXCR5<SUP>+</SUP>, CD4<SUP>+</SUP>ICOS<SUP>+</SUP>, and CD4<SUP>+</SUP>PD-1<SUP>+</SUP> cells. The Ascl2 level was higher in peripheral blood mononuclear cells from SS patients compared with those from healthy controls. Our findings suggest that Ascl2 may play a role in the development and progression of SS and may be a therapeutic target in the treatment of SS.

Growth inhibitory effects of obovatol through induction of apoptotic cell death in prostate and colon cancer by blocking of NF-κB

Lee, S.Y.,Yuk, D.Y.,Song, H.S.,Yoon, D.Y.,Jung, J.K.,Moon, D.C.,Lee, B.S.,Hong, J.T. North-Holland ; Elsevier Science Ltd 2008 european journal of pharmacology Vol.582 No.1

Biphenolic components in Magnolia obovata including magnolol and honokiol have shown several pharmacological activities such as anti-tumor, anti-oxidant and anti-inflammatory effects. Previously in cultured macrophage Raw264.7 cells and fibroblast, we found that obovatol, an active compound isolated from M. obovata inhibited NF-κB activity which has been known to be a significant transcriptional factor to control of cancer cell growth. We investigated here whether obovatol could inhibit NF-κB activity, and thereby inhibit cancer cell growth in prostate (LNCaP and PC-3) and colon cancer (SW620 and HCT116) cells. Treatment of obovatol (10, 15, 20, 25 μM) inhibits cancer cell growth in the absence or the presence of tumor necrosis factor-α (TNF-α , 10 ng/ml) and tetradecanoyl phorbol acetate (TPA 10 or 50 nM) in a concentration-dependent manner through induction of apoptotic cell death. Cytotoxic activity was not observed in normal cells with up to 50 μM obovatol. It was also found that obovatol inhibited TNF-α and TPA-induced transcriptional and DNA binding activities of NF-κB. In further study, obovatol decreased translocation p65 and p50 into nucleus via decrease of phosphorylation of IκB. Correlated well with the induction of apoptosis, obovatol increased the expression of the apoptotic genes; Bax, caspase-3, caspase-9, whereas inhibited expression of anti-apoptotic genes; Bcl-2, inhibitor of apoptosis protein (IAP-1) and X chromosome IAP (XIAP) as well as the cell proliferation marker genes; Cox-2, c-Fos, c-Jun and cyclin D1. These results suggest that obovatol inhibits prostate and colon cancer cell growth via induction of apoptotic cell death, and that inhibition of NF-κB may be a significant as its action mechanism.

CK2α/CSNK2A1 Phosphorylates SIRT6 and Is Involved in the Progression of Breast Carcinoma and Predicts Shorter Survival of Diagnosed Patients

Bae, J.S.,Park, S.H.,Jamiyandorj, U.,Kim, K.M.,Noh, S.J.,Kim, J.R.,Park, H.J.,Kwon, K.S.,Jung, S.H.,Park, H.S.,Park, B.H.,Lee, H.,Moon, W.S.,Sylvester, K.G.,Jang, K.Y. American Association of Pathologists and Bacteriol 2016 The American journal of pathology Vol.186 No.12

<P>Recently, the roles of sirtuins (SIRTs) in tumorigenesis have been of interest to oncologists, and protein kinase CK2 alpha 1 (CSNK2A1) has been shown to be involved in tumorigenesis by phosphorylating various proteins, including SIRT1. Therefore, we evaluated the roles of CSNK2A1, SIRT6, and phosphorylated SIRT6 and their relationships in breast carcinoma. Nuclear expression of CSNK2A1 and SIRT6 predicted shorter overall survival and relapse-free survival by multivariate analysis. Inhibition of CSNK2A1 decreased the proliferative and invasive activity of cancer cells. In addition, CSNK2A1 was bound to SIRT6 and phosphorylated SIRT6; evidence for this is provided from immunofluorescence staining, co-immunoprecipitation of CSNK2A1 and SIRT6, a glutathione S-transferase pull-down assay, an in vitro kinase assay, and transfection of mutant CSNK2A1. Knockdown of SIRT6 decreased the proliferation and invasiveness of cancer cells. Overexpression of SIRT6 increased proliferation, but mutation at the Ser338 phosphorylation site of SIRT6 inhibited the proliferation of MCF7 cells. Moreover, both knockdown of SIRT6 and a mutation at the phosphorylation site of SIRT6 decreased expression of matrix metallopeptidase 9, beta-catenin, cyclin D1, and NF-kappa B. Especially, SIRT6 expression was associated with the nuclear localization of B-catenin. This study demonstrates that CSNK2A1 and SIRT6 are indicators of poor prognosis for breast carcinomas and that CSNK2A1-mediated phosphorylation of SIRT6 might be involved in the progression of breast carcinoma.</P>

Caspase-3 activation as a key factor for HBx-transformed cell death

Kim, A.,Kwon, O. S.,Kim, S. O.,He, L.,Bae, E. Y.,Lee, M. S.,Jeong, S. J.,Shim, J. H.,Yoon, D. Y.,Kim, C. H.,Moon, A.,Kim, K. E.,Ahn, J. S.,Kim, B. Y. Blackwell Publishing Ltd 2008 Cell proliferation Vol.41 No.5

<P>Abstract. </P><P><I>Objectives</I>: Nuclear factor-kappa B (NF-&kgr;B) activation has been associated with the tumorigenic growth of hepatitis B virus X protein (HBx)-transformed cells. This study was aimed to find a key target for treatment of HBx-mediated cancers. <I>Materials and methods</I>: NF-&kgr;B activation, endoplasmic reticulum-stress (ER-stress), caspase-3 activation, and cell proliferation were evaluated after Chang/HBx cells permanently expressing HBx viral protein were treated with inhibitors of NF-&kgr;B, proteasome and DNA topoisomerase. <I>Results</I>: Inhibition of NF-&kgr;B transcriptional activity by transient transfection with mutant plasmids encoding Akt1 and glycogen synthase kinase-3&bgr; (GSK-3&bgr;), or by treatment with chemical inhibitors, wortmannin and LY294002, showed little effect on the survival of Chang/HBx cells. Furthermore, I&kgr;Bα (S32/36A) mutant plasmid or other NF-&kgr;B inhibitors, 1-pyrrolidinecarbonidithioic acid and sulphasalazine, were also shown to have little effect on the cell proliferation. By contrast, proteasome inhibitor-1 (Pro1) and MG132 enhanced the HBx-induced ER-stress response and the subsequent activation of caspase-12, -9 and -3 and reduced cell proliferation. Camptothecin (CPT), however, triggered activation of caspase-3 without induction of caspase-12, and reduced cell proliferation. In addition, CPT-induced cell death was reversed by pre-treatment with z-DEVD, a caspase-3-specific inhibitor. <I>Conclusions</I>: Detailed exploitation of the regulators of caspase-3 activation could open the gate for finding an efficient target for development of anticancer therapeutics against HBx-transformed hepatocellular carcinoma.</P>

문학반응이론에 의한 보건의학기술계열 국가시험개선방안에 관한 연구

이창규,이승관,조경진,박종성,정수경,유병서,박상숙,윤효숙,황선철,문경환,김정민,함용운,김지환,임국환,김영순,윤경희,황성준 高麗大學校 倂設 保健大學 保健科學硏究所 1998 保健科學論集 Vol.24 No.1

The nationally-governed examinations for certification of allied health professions in Korea have been continued for thirty three years. During that time, there were a lot of managerial improvements in carrying out the examinations, for example, the looking-over the papers converted from manual method to computerized one. Nevertheless, the overall aspects of item management in the national examinations are still remained as obsolete style. In some developed countries they have already tried or adopted computerized system in making questions, executing item analysis, developing item banking, and in overall management of examinations, looking over papers for their national licensure examinations, and have established good reputations. Since the National Health Personnel Licensing Examination Board was established in 1998, now we can expect there would be a lots of improvements in the managerial systems and organizational structures associated with the national licensing examinations suggesting followings. 1. The contents and scope of the licensing examinations based on job analysis should be announced publicaly. 2. Items should be developed based on the scopes and contents of job characteristics. 3. All the developed items for the licensing examinations should be tested quantitatively prior to banking items. 4. All the parameters of the stored items should be fully estimated through item response theory. 5. All the efforts should be given in order to execute the examinations in individual residential areas on behalf of every examinee. 6. To the licensing examination system the Computerized Adaptive Testing system should be introduced in order to enhance the efficiencies. 7. Security enhancement on the stored items should be reminded in order to prevent leaking out the banking items. 8. Much more improvements should be made for the proper job evaluations. 9. Every efforts should be given to prevent cheatings encountable during the examinations. 10. Make the best of professional volunteers from the various fields in the execution of licensing examinations. 11. Consider a new system in that examinations can be executed twice or more in a year. 12. The current methods of presiding over the examinations should be replaced by a more reasonable one. 13. The results of the examinations should be announced as soon as possible in order not to prolong the examinees' unemployment periods. 14. The National Health Personnel Licensing Examination Board should try to rationalize the management keeping step with the information-oriented society.

식품의 제조 가공용 화학적 합성품의 국내 소비실태에 관한 연구

문범수,김복성,이재관,윤혜,신석우 國立保健硏究院 1972 국립보건연구원보 Vol.9 No.-

食品中의 Nitrosamine에 關한 硏究(第1報) : 1. 食品中의 窒酸鹽 및 亞窒酸鹽의 含量 Contents of Nitrate and Nitrite in Various Foods

文範洙,金福成,李載寬,禹相奎 國立保健硏究院 1973 국립보건연구원보 Vol.10 No.-