• Biomolecules & Therapeutics
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• v.6 no.3
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• pp.276-282
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• 1998

Retinoids regulate a wide variety of biological processes such as cellular proliferation and differentiation in many cell types. They have also shown to stimulate replication of several viruses including human cytomegalovirus (CMV). Retinoid signalling pathway involves two distinct subfamilies of nuclear receptors, retinoic acid receptors (RARs) and retinoid X receptors (RXRs) that bind to specific retinoic acid response elements (RAREs) in the promoter regions of retinoid-target genes. Here, we characterized RAREs in the regulatory regions of the CMV and of the hepatitis B vi.us (HBV). The viral RAREs, i.e., CMV-RARE and HBV-RARE, are composed of two consensus RARE half-sites (A/GGGTCA) arranged as a direct repeat separated by 5-bp and 1-bp, respectively. The RAREs were activated by both RAR/RXR heterodimers and RXR homodimers in transient transfection experiments. We also found that COUP-TF$\alpha$ (chicken ovalbumin upstream promoter-transcription factor u) and COUP-TF$\beta$ repressed the retinoid response of the viral elements. Further we demonstrated that previously known retinoid antagonist, SRI 1330, repressed retinoid-induced transactivation of the CMV-RARE. These results implicate Vitamin A, it's nuclear receptors and COUP-TFs as important regulators of the CMV and HBV pathogenesis and the SRl1330 as potential negative modulator of such retinoid-dependent processes.

• Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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• 2013.10a
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• pp.946-948
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• 2013

Several baculovirus vector systems recombined with coding genes of polyhedron promoter, vesicular stomatitis virus G (VSVG), polyA, cytomegalovirus (CMV) promoter, enhanced green fluorescent protein (EGFP), and protein transduction domain (PTD) were constructed. These recombinant baculovirus vector systems were applied into human foreskin fibroblast cells and compared the effects of gene transfer and gene expression of these recombinant baculovirus vector systems with control vector system. From this study, it showed that these novel recombinant baculovirus vector systems were superior efficacy to control vector system in view of gene transfer and gene expression.

• YAKHAK HOEJI
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• v.50 no.4
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• pp.268-271
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• 2006

Protein kinase UL97 is an unusual protein kinase that can phosphorylate nucleoside analogs as well as protein/peptide. Previously we found a H2B-derived peptide, KESYSVYVYKV and reported that the P+5 position (K) is important. To further understand the substrate specificity at the P+5 position, we introduced spot assay system and showed that a peptide containing K residue among other amino acids at the P+5 position is the best substrate. Also other residues such as M, I, L, or G are good enough to be substrate of UL97. This result may aid the discovery of a new antiviral inhibitor.

• Journal of Microbiology
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• v.39 no.3
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• pp.226-228
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• 2001

Human lymphotropic herpesvirus is known to be a major pathogen associated with various diseases in bone marrow transplantation (BMT) recipients. A multiplex nested-polymerase chain reaction (PCR) method was developed for the simultaneous detection of human lymphotropic herpesviruses, including Ebstein-Barr virus (EBV), cytomegalovirus (CMV), and human herpesvirus 6 variants A and B (HHV6-A, HHV6-B). To demonstrate the usefulness of multiplex PCR for the analysis of clinical samples, peripheral blood mononuclear cells and serum from BMT recipients were analysed. The results skewed that a clear detection could be made between EBV, HCMV and HHV-6. This multiplex PCR assay is an efficient and cost-effective approach to the analysis of large numbers of samples to determine the epidemiological importance of EBV HCMV and HHV-6.

• Annals of Clinical Neurophysiology
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• v.7 no.1
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• pp.25-27
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• 2005

The Guillain-Barre syndrome (GBS) is an acute polyradiculoneuropathy marked by flaccid areflexic paralysis. Although the pathogenesis of GBS remains incompletely defined, considered as an autoimmune disease most frequently triggered by an previous infection. Antecedent infections with Campylobacter jejuni, cytomegalovirus, Ebstein-Barr virus, Mycoplasma pneumoniae, Haemophilus influenzae, human immunodeficiency virus, enterovirus, rotavirus are common. But, it is rare that GBS following typhoid fever. We present a case of typical GBS after antecedent Salmonella typhi infection.

• YAKHAK HOEJI
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• v.52 no.6
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• pp.466-470
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• 2008

Human cytomegalovirus expresses an unusual protein kinase UL97, a member of ${H_V}{U_L}$ family of protein kinase. UL97 can phosphorylate nucleoside analogs such as ganciclovir as well as protein/peptide. It has previously been reported that UL97 is able to phosphorylate a KESYSVYVYKV peptide and that P+5 position (K) is important. We examined the extent of contribution of other positions (P-4 through P+6) of the peptide to be substrate of UL97 using alanine substituted peptides (Ala scanning) and deleted peptides. The result suggested that the E (P-2) is negative effect and P+5 (K) is still important. The peptide YSVYVYK is the shortest substrate enough to show high activity, which could be a starting point to develop peptidomimetic drug. This study would give important information to deeply understand the substrate specificity of UL97 and develop an antiviral drug using the small peptide identified here.

• Korean Journal of Agricultural Science
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• v.48 no.2
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• pp.353-365
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• 2021

Previously, we reported that tissue factor (Tf) was included in the list of differentially expressed genes as an upregulated gene in a rejected porcine heart after xenotransplantation into monkey. In this study, we analyzed that expression of Tf in aortic endothelial cells (pAEC) isolated from alpha 1,3-galactosyltransferase knockout pig in response to allogeneic porcine serum and xenogeneic human serum. The consequence was significant upregulation of Tf expression by responding to human serum compared with porcine serum. To analyze the function of Tf gene as a promoter, we constructed reporter vectors for expression of luciferase linked to 1,246 and 787 base pairs of porcine Tf (pTF1246 and pTF787), and 535 base pairs of human TF (hTF535) sequences including putative promoter regions and AP-1 biding site at the 5' end. The reporter vectors were transfected into pAEC including cytomegalovirus enhancer/chicken β-actin (CAG)-luciferase vector as a control. Luciferase assay showed that all of the promoters were insufficient to express luciferase compared with CAG promoter in basic culture conditions. Notably, pTF1246, pTF787, and hTF535 led to a significant increase of luciferase expression in response to human serum compared with porcine serum while no change of CAG. pTF1246 and pTF787 showed higher expression than hTF535. Taken together, our findings suggest that pTF1246 and pTF787 promoters could mediate target gene expression specifically at xenogeneic stress condition.

• Pediatric Infection and Vaccine
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• v.6 no.2
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• pp.253-260
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• 1999

Purpose : The purpose of this study is to make and use monoclonal Ab which reacts with CMV major immediate early(${\alpha}$) protein(p72). Methods : Normal human fibroblast(Foreskin derived) was cultured in Eagle's minimal essential medium(MEM) containing 10% cowfetus serum and mouse chondroblast was cultured in P3X63 Ag8.653(ATCC. Maryland USA) to maintain $5{\times}10^5/ml$ cell counts. CMV(KJHJ90) from congenital CMV infected infant's urine was multiplied and used for Ab making. CMV Ag was injected 4 times, 1 week interval into the peritoneal space of 6~8 weeks old mice. And then lymphocyles and fibroblasts taken from spleen were obtained and conjugated. After the conjugated cell cultured, we chose the cell that had high Ab titer using indirect immunofluerescent method. Results : Among the 28 monoclonal antibodies obtained LPC12 and LPC23 reacted highly with nucleus of AD169 infected cell. Purified AD169 after SDS-PAGE, molecular weight of Ag, which reacted with purified monoclonal Ab, was obtained using Western blotting. Monoclonal Ab of LPC12 and LPC23 clone reacted most highly with 72 kd Ag. Conclusion : LPC12 and LPC23 clonal Ab with AD 169(P63-27) is useful on early diagnosis of CMV infection.

• Reproductive and Developmental Biology
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• v.28 no.3
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• pp.191-196
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• 2004

In this study, to construct more effective retrovirus system, we compared four internal promoters (RSV: Rous sarcoma virus, UbC: Ubiquitin C, β-actin, CMV: human cytomegalovirus) in the retrovirus vector by measuring GFP (green fluorescent protein) expression. The effect of WPRE (woodchuck hepatitis virus posttrans-criptional regulatory element) sequence on transgene expression was also investigated. Experiments were conducted with cells derived from three different species (human, pig and chicken) and evaluated the activity of each promoter and the effect of WPRE sequence by fluorometry and Western blotting. In all cells tested, RSV and CMV promoters were superior to UbC and β-actin promoters, and RSV promoter was the best one in chicken cells. The boosting effect of WPRE sequence on GFP expression was evident in human and porcine cells but not in chicken cells.

• The Korean Journal of Physiology and Pharmacology
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• v.3 no.6
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• pp.597-603
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• 1999

Caldesmon (CaD), one of microfilament-associated proteins, plays a key role in microfilament assembly in mitosis. We have investigated the effects of overexpression of the high molecular weight isoform of CaD (h-CaD) on the physiology of vascular smooth muscle cells (VSMCs). Rat aortic VSMCs were stably transfected with plasmids carrying a full length human h-CaD cDNA under control of cytomegalovirus promoter. The majority of the overexpressed h-CaD appears to be localized predominantly on cytoskeleton structures as determined by detergent lysis. The overexpression of h-CaD, however, does not decrease the level of endogenous low molecular weight isoform of CaD. h-CaD overexpressing VSMCs (h-CaD/VSMCs) show a decreased growth rate than that of vector-only transfected cells when determined by $[^3H]thymidine$ uptake and cell counting after fetal bovine serum (FBS) stimulation. h-CaD/VSMCs were smaller than vector-transfected cells by 18% in cell diameter. These data suggest that overexpression of h-CaD can inhibit the poliferation and the cell volume of VSMCs stimulated by growth factors and that the gene therapy with h-CaD may be helpful to prevent the conditions associated with hypertrophy and/or hyperplasia of VSMCs after arterial injuries.