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Bae, Yoonhee,Rhim, Hyang-Shuk,Lee, Seulgi,Ko, Kyung Soo,Han, Jin,Choi, Joon Sig John Wiley & Sons 2017 Journal of Pharmaceutical Sciences Vol.106 No.6
<P>Malignant glioma is the most common and aggressive form of primary brain tumor in adults. In this study, we describe the efficacy of nonviral gene delivery carriers, histidine-and arginine-or histidine- and lysine-grafted polyamidoamine (PAMAM) dendrimers (PAMAM-H-R and PAMAM-H-K), in delivering a therapeutic and a tumor-selective killer gene, apoptin, using human glioma cells (U87-MG) and newborn human dermal fibroblast cells. We analyzed transfection efficiency using luciferase and a plasmid DNA encoding for enhanced green fluorescent protein and assessed cell viability in both cells. The results show that transfection efficiency of PAMAM-H-R and PAMAM-H-K was greatly increased compared with that of native PAMAM. Moreover, among PAMAM derivatives, cytotoxicity of PAMAM-H-K was very low. We treated both cells with complexes of PAMAM-H-R or PAMAM-H-K and apoptin and analyzed their cellular uptake by flow cytometry and localization by confocal microscopy. Furthermore, cell cycle distribution, caspase 3 activity assay, and JC-1 analysis showed cell death induced by apoptin in U87-MG cells. The present study demonstrates that a PAMAM-H-R/apoptin complex is an effective gene carrier system in glioma cell culture. (C) 2017 American Pharmacists Association (R). Published by Elsevier Inc. All rights reserved.</P>
Choi, Soo Young,Rhim, Hyang Shuk,Park, Jin Seu,Lee, Han Gyu,Lee, Yoon,Kang, Young Hee 생화학분자생물학회 2001 BMB Reports Vol.33 No.4
The human immunodeficiency virus type 1 (HIV-1), transactivator of transcription (Tat), is one of the viral gene products that is essential for HIV 1 replication. The HIV-1 Tat protein regulates transcription from an HIV-1 long terminal repeat (LTR) and affects the gene expression of cellular proteins during infection. In order to develop an expression system to overexpress and simply purify HIV-1 Tat proteins, the HIV-1 Tat coding sequences that contain one or two exons were amplified using PCR and cloned into a pET vector, which contains a consecutive stretch of six histidine residues at the amino-terminus. The reconstituted vectors were overexpressed in the E. coli strain and the soluble recombinant proteins were purified to be homogeneity in a single step by Ni^(+2)-nitrilotriacetic acid Sepharose chromatography under nondenaturing conditions. Recombinant HIV-1 Tat proteins were shown to transactivate the HIV-1 LTR promoter in a dose-dependent manner when introduced into mammalian cells. In addition, treatment of human endothelial cells with purified Tat proteins resulted in a significant increase in the level of vascular cell adhesion molecule-1 (VCAM-1) expression. These results indicate that the recombinant HIV-1 Tat proteins are active in transactivating viral and cellular promoters. The expression and purification system described in this study will facilitate in characterizing the biological functions of the Tat proteins.
Anti-complement Effects of Anion-Substituted Poly(vinyl alcohol) Membranes
Ryu, Kyu-Eun,Rhim, Hyang-Shuk,Park, Chong-Won,Chun, Heung-Jae,Hong, Seung-Hwa,Kim, Young-Chai,Lee, Young-Moo The Polymer Society of Korea 2004 Macromolecular Research Vol.12 No.1
In a continuation of our previous studies on blood compatibility profiles of anion-substituted poly(vinyl alcohol) (PVA) membranes, in which hydroxyl groups have been replaced with carboxymethyl (C-PVA) and sulfonyl groups (S-PVA), we have studied the activation of complement components and the changes in white cell and platelet count in vitro and compared them with those of unmodified PVA, Cuprophane, and low-density polyethylene. Complement activation of fluid phase components, C3a, Bb, iC3b, and SC5b-9, and of bound phases, C3c, C3d, and SC5b-9, were assessed by enzyme-linked immunosorbent assay (ELISA) and immunoblot, respectively. The changes in the number of white cells and platelets following complement activation were counted using a Coulter counter. C-PVA and S-PVA activated C3 to a lesser extent than did PVA, which we attribute to the diminished level of surface nucleophiles of the samples. In addition, C- and S-PVA exhibit increased inhibition of Bb production, resulting in a decrease in the extent of C5 activation. Consequently, because of the reduced activation of C3 and C5, C- and S-PVA samples cause marked decreases in the SC5b-9 levels in plasma. We also found that the negatively charged sulfonate and carboxylate groups of the samples cause a greater extent of adsorbtion of the positively charged anaphylatoxins, C3a and C5a, because of strong electrostatic attraction, which in turn provides an inhibition of chemotaxis and activation of leukocytes. The ability to inhibit complement production, together with the binding ability of anaphylatoxins of the C- and S-PVA samples, leads to a prominent decrease in lysis of leukocytes as well as activation of platelets.
Woo, Dong-Cheol,Kim, Goo-Young,Kim, Hyun-Ju,Bang, Eunjung,Rhim, Hyang-Shuk,Kim, Sang-Young,Lee, Do-Wan,Choi, Chi-Bong,Seoung, Youl-Hun,Choe, Bo-Young Hindawi Limited 2013 Journal of spectroscopy Vol.2013 No.-
<P>The purpose of this study is to investigate the metabolic alterations associated with acute alcohol treatment in zebrafish by<SUP>1</SUP>H nuclear magnetic resonance spectroscopy (NMRS). The brain metabolism of zebrafish was investigated after acute alcohol treatment (one-hour long exposure of adult fish to 0.00%, 0.25%, 0.50%, or 1.00% ethyl alcohol) with whole brain extraction. The results of this study showed that glutamate (Glu) was significantly decreased, scyllo-inositol (sIns) showed a small apparent increase only in the highest acute treatment dose group, and myoinositol (mIns) showed a significant decrease. [Glu]/[tCr] and [mIns]/[tCr] levels were significantly reduced regardless of the alcohol dose, and [sIns]/[tCr] was increased in the highest alcohol treatment dose group. The present NMR study revealed that specific metabolites, such as Glu and mIns, were substantially decreased in case of acute alcohol exposed zebrafish brain.</P>