VP2 capsid domain of the H-1 parvovirus determines susceptibility of human cancer cells to H-1 viral infection

Cho, I-R,Kaowinn, S,Song, J,Kim, S,Koh, S S,Kang, H-Y,Ha, N-C,Lee, K H,Jun, H-S,Chung, Y-H Nature America, Inc. 2015 Cancer gene therapy Vol.22 No.5

Although H-1 parvovirus is used as an antitumor agent, not much is known about the relationship between its specific tropism and oncolytic activity. We hypothesize that VP2, a major capsid protein of H-1 virus, determines H-1-specific tropism. To assess this, we constructed chimeric H-1 viruses expressing Kilham rat virus (KRV) capsid proteins, in their complete or partial forms. Chimeric H-1 viruses (CH1, CH2 and CH3) containing the whole KRV VP2 domain could not induce cytolysis in HeLa, A549 and Panc-1 cells. However, the other chimeric H-1 viruses (CH4 and CH5) expressing a partial KRV VP2 domain induced cytolysis. Additionally, the significant cytopathic effect caused by CH4 and CH5 infection in HeLa cells resulted from preferential viral amplification via DNA replication, RNA transcription and protein synthesis. Modeling of VP2 capsid protein showed that two variable regions (VRs) (VR0 and VR2) of H-1 VP2 protein protrude outward, because of the insertion of extra amino-acid residues, as compared with those of KRV VP2 protein. This might explain the precedence of H-1 VP2 protein over KRV in determining oncolytic activity in human cancer cells. Taking these results together, we propose that the VP2 protein of oncolytic H-1 parvovirus determines its specific tropism in human cancer cells.

돼지 H-FABP 유전자의 다형성 및 경제 형질과의 연관성 구명

최봉환,김태헌,이지웅,조용민,이혜영,조병욱,정일정 한국동물자원과학회 2003 한국축산학회지 Vol.45 No.5

The purpose of this study was to detect association between genetic variation and economic trait in the porcine heart type fatty acid-binding protein gene as a candidate gene for the traits related with growth and meat quality in pigs. The H-FABP is a 15-kDa protein expressed in several tissues with high demand for fat metabolism such as cardiac and skeletal muscle and lactating mammary gland. H-FABP is small intracellular protein involved in fatty acid transport from the plasma membrane to the site of β-oxidation and/or triacylglycerol or phospholipid synthesis. In this study, H-FABP PCR-RFLP was performed in F_(2) population composed of 214 individuals form an intercross between Korean Native Boars and Landrace sows. PCR products form tow primer sets within H-FABP gene were amplified in 850bp and 700bp. Digestion of PCR products with the restriction digestion enzymes HaeⅢ and Hinf Ⅰ, revealed fragment length polymorphisms(RFL. Ps). The genotype frequencies from H-FABP/HaeⅢ was .29 for genotype DD, .53 for genotype Dd, and .15 for genotype dd, respectively. The genotype frequencies of HH, Hh, and hh from H-FABP(hinf Ⅰ was .38, .41, and .20, respectively, in the population.Relationships between their genotypes and economic traits were estimated. In H-FABP/HaeⅢ locus, there were specific genotypes(Dd and dd) associated with economic traits such as body weight. In H-FABP/Hinf Ⅰ Iocus, Genotypes of HH and Hh associated with growth traits such as body weights at 5, 12, and 30 week of age (p<.05 or p<.001) and back fat thickness, body fat including abdominal and trimmed fat (p<.001) and intramuscular fat(p<.05). The 'H'allele was positivecly associated with gaining of body weight and fatness deposition. In conclusion, a significant association of the H-FABP gene from its genetic variation was found on body weight, intramuscular fat and backfat thickness.

Supplementation of oil-based inactivated H9N2 vaccine with M2e antigen enhances resistance against heterologous H9N2 avian influenza virus infection

Park, J.K.,Lee, D.H.,Cho, C.H.,Yuk, S.S.,To, E.O.,Kwon, J.H.,Noh, J.Y.,Kim, B.Y.,Choi, S.W.,Shim, B.S.,Song, M.K.,Lee, J.B.,Park, S.Y.,Choi, I.S.,Song, C.S. Elsevier Scientific Pub. Co 2014 Veterinary microbiology Vol.169 No.3

Avian influenza virus (AIV) subtype H9N2 has been evolving rapidly and vaccine escape variants have been reported to cause circulation of infections and economic losses. In the present study, we developed and evaluated ectodomain of the AIV matrix 2 (M2e) protein as a supplementing antigen for oil-based inactivated H9N2 vaccine to increase resistance against vaccine escape variants. AIV H9N2 M2e antigen was expressed in Escherichia coli and supplemented to inactivated H9N2 oil emulsion vaccine. Specific pathogen-free chickens received a single injection of inactivated H9N2 oil emulsion vaccines with or without M2e supplementation. At three weeks post vaccination, hemagglutination inhibition tests and enzyme-linked immunosorbent assays were performed to determine serological immune responses. Challenge study using a vaccine escape H9N2 variant was performed to evaluate the efficacy of M2e supplementation. M2e antigen supplemented in oil emulsion vaccine was highly immunogenic, and a single M2e-supplemented vaccination reduced challenge virus replication and shedding more effectively than non-supplemented vaccination.

Coumarins reduce biofilm formation and the virulence of Escherichia coli O157:H7

Lee, J.H.,Kim, Y.G.,Cho, H.S.,Ryu, S.Y.,Cho, M.H.,Lee, J. G. Fischer 2014 Phytomedicine Vol.21 No.8

E. coli O157:H7 is the most common cause of hemorrhagic colitis, and no effective therapy exists for E. coli O157:H7 infection. Biofilm formation is closely related to E. coli O157:H7 infection and constitutes a mechanism of antimicrobial resistance. Hence, the antibiofilm or antivirulence approach provides an alternative to antibiotic strategies. Coumarin and its derivatives have a broad range of biological effects, and in this study, the antibiofilm activities of nine coumarins were investigated against E. coli O157:H7. Coumarin or umbelliferone at 50μg/ml was found to inhibit biofilm E. coli O157:H7 formation by more than 80% without affecting bacterial growth. Transcriptional analysis showed that coumarins repressed curli genes and motility genes in E. coli O157:H7, and these findings were in-line with observed reductions in fimbriae production, swarming motility, and biofilm formation. In addition, esculetin repressed Shiga-like toxin gene stx2 in E. coli O157:H7 and attenuated its virulence in vivo in the nematode Caenorhabditis elegans. These findings show that coumarins have potential use in antivirulence strategies against persistent E. coli O157:H7 infection.

Simultaneous subtyping and pathotyping of the novel reassortant influenza A (H5N8) virus from clinical samples using a diagnostic microarray

Kwon, J. H.,Kim, J. H.,Lee, D. h.,Cho, H.,Hwang, S. Y.,Yuk, S. S.,Erdene-Ochir, T. O.,Noh, J. Y.,Hong, W. T.,Jeong, J. H. Springer Science + Business Media 2016 BioChip Journal Vol.10 No.3

<P>Highly pathogenic avian influenza (HPAI) viruses cause economic losses in the poultry industry and pose a severe threat to human health. Rapid and accurate diagnostic methods are important because they can help prevent further spread of the virus and reduce the time required for eradication of the virus. We developed a low-density microarray for the rapid detection and identification of avian influenza virus subtypes H5, H7, and H9 and their pathotypes in a previous study. In the present study, we report the development of updated probe sets and evaluation of the diagnostic microarray using H5N8 clade 2.3.4.4 HPAI viruses including clinical samples, without the need for egg propagation. Cy3-labeled DNA targets were obtained by reverse transcription polymerase chain reaction using Cy3-labeled universal primers, and labeled amplicons were hybridized to the microarray. All positive samples from RT-PCR showed H5-specific and highly pathogenic pattern in the microarray, without purification of PCR products. Furthermore, it allowed for specific detection of the subtype and pathotype from low DNA concentration samples that did not allow direct sequence analysis. Therefore, this diagnostic microarray has enormous potential for the rapid subtyping and pathotyping from clinical samples without the need for culture.</P>

Tat-PRAS40 prevent hippocampal HT-22 cell death and oxidative stress induced animal brain ischemic insults

Shin, M.J.,Kim, D.W.,Jo, H.S.,Cho, S.B.,Park, J.H.,Lee, C.H.,Yeo, E.J.,Choi, Y.J.,Kim, J.A.,Hwang, J.S.,Sohn, E.J.,Jeong, J.H.,Kim, D.S.,Kwon, H.Y.,Cho, Y.J.,Lee, K.,Han, K.H.,Park, J.,Eum, W.S.,Choi, Pergamon ; Elsevier Science Ltd 2016 FREE RADICAL BIOLOGY AND MEDICINE Vol.97 No.-

Proline rich Akt substrate (PRAS40) is a component of mammalian target of rapamycin complex 1 (mTORC1) and is known to play an important role against reactive oxygen species-induced cell death. However, the precise function of PRAS40 in ischemia remains unclear. Thus, we investigated whether Tat-PRAS40, a cell-permeable fusion protein, has a protective function against oxidative stress-induced hippocampal neuronal (HT-22) cell death in an animal model of ischemia. We showed that Tat-PRAS40 transduced into HT-22 cells, and significantly protected against cell death by reducing the levels of H<SUB>2</SUB>O<SUB>2</SUB> and derived reactive species, and DNA fragmentation as well as via the regulation of Bcl-2, Bax, and caspase 3 expression levels in H<SUB>2</SUB>O<SUB>2</SUB> treated cells. Also, we showed that transduced Tat-PARS40 protein markedly increased phosphorylated RRAS40 expression levels and 14-3-3σ complex via the Akt signaling pathway. In an animal ischemia model, Tat-PRAS40 effectively transduced into the hippocampus in animal brain and significantly protected against neuronal cell death in the CA1 region. We showed that Tat-PRAS40 protein effectively transduced into hippocampal neuronal cells and markedly protected against neuronal cell damage. Therefore, we suggest that Tat-PRAS40 protein may be used as a therapeutic protein for ischemia and oxidative stress-induced brain disorders.

Identification of novel scaffolds for potential anti-Helicobacter pylori agents based on the crystal structure of H. pylori 3-deoxy-d-manno-octulosonate 8-phosphate synthase (HpKDO8PS)

Cho, S.,Im, H.,Lee, K.Y.,Chen, J.,Kang, H.J.,Yoon, H.J.,Min, K.H.,Lee, K.R.,Park, H.J.,Lee, B.J. S.E.C.T. [etc.] ; Elsevier Science Ltd 2016 European journal of medicinal chemistry Vol.108 No.-

The crystal structure of 3-deoxy-d-manno-octulosonate-8-phosphate synthase (KDO8PS) from Helicobacter pylori (HpKDO8PS) was determined alone and within various complexes, revealing an extra helix (HE) that is absent in the structures of KDO8PS from other organisms. In contrast to the metal coordination of the KDO8PS enzyme from Aquifex aeolicus, HpKDO8PS is specifically coordinated with Cd<SUP>2+</SUP> or Zn<SUP>2+</SUP> ions, and isothermal titration calorimetry (ITC) and differential scanning fluorimetry (DSF) revealed that Cd<SUP>2+</SUP> thermally stabilizes the protein structure more efficiently than Zn<SUP>2+</SUP>. In the substrate-bound structure, water molecules play a key role in fixing residues in the proper configuration to achieve a compact structure. Using the structures of HpKDO8PS and API [arabinose 5-phosphate (A5P) and phosphoenolpyruvate (PEP) bisubstrate inhibitor], we generated 21 compounds showing potential HpKDO8PS-binding properties via in silico virtual screening. The capacity of three, avicularin, hyperin, and MC181, to bind to HpKDO8PS was confirmed through saturation transfer difference (STD) experiments, and we identified their specific ligand binding modes by combining competition experiments and docking simulation analysis. Hyperin was confirmed to bind to the A5P binding site, primarily via hydrophilic interaction, whereas MC181 bound to both the PEP and A5P binding sites through hydrophilic and hydrophobic interactions. These results were consistent with the epitope mapping by STD. Our results are expected to provide clues for the development of HpKDO8PS inhibitors.

Sodium-ion supercapacitors based on nanoporous pyroproteins containing redox-active heteroatoms

Cho, S.Y.,Yoon, H.J.,Kim, N.R.,Yun, Y.S.,Jin, H.J. Elsevier Sequoia 2016 Journal of Power Sources Vol.329 No.-

Nanostructured carbon-based materials fabricated via simple methods from renewable bio-resources have great potential in rechargeable energy storage systems. In this study, nanoporous pyroproteins containing a large amount of redox-active heteroatoms (H-NPs) were fabricated from silk fibroin by an in situ carbonization/activation method. The H-NPs have a large surface area of ~3050 m<SUP>2</SUP> g<SUP>-1</SUP>, which is mainly comprised of nanometer-scale pores. Also, these H-NPs have oxygen and nitrogen heteroatoms of 17.4 wt% and 2.9 wt%, respectively. Synergistic sodium ion storage behaviors originate from electrochemical double layer capacitance and pseudocapacitance, leading to very high electrochemical performances of H-NPs in aqueous and non-aqueous electrolyte systems. Sodium-ion supercapacitors (NISs) based on commercial graphite//H-NPs show a high specific power of ~1900 W kg<SUP>-1</SUP> at ~77 Wh kg<SUP>-1</SUP>. Also, NISs based on commercial hard carbon//H-NPs exhibit a high specific energy of ~217 Wh kg<SUP>-1</SUP> at ~42 W kg<SUP>-1</SUP>. In addition, outstanding cycling performances over 30,000 cycles are achieved for symmetric NISs.

Improvement of hydrogenated amorphous silicon germanium thin film solar cells by different p-type contact layer

Cho, J.,Pham, D.P.,Jung, J.,Shin, C.,Park, J.,Kim, S.,Tuan Le, A.H.,Park, H.,Iftiquar, S.M.,Yi, J. Pergamon 2016 Materials science in semiconductor processing Vol.41 No.-

In this study, we report an appreciably increased efficiency from 6% up to 9.1% of hydrogenated amorphous silicon germanium (a-SiGe:H) thin film solar cells by using a combination of different p-doped window layers, such as boron doped hydrogenated amorphous silicon (p-a-Si:H), amorphous silicon oxide (p-a-SiO<SUB>x</SUB>:H), microcrystalline silicon (p-@?c-Si:H), and microcrystalline silicon oxide (p-@?c-SiO<SUB>x</SUB>:H). Optoelectronic properties and the role of these p-layers in the enhancement of a-SiGe:H cell efficiency were also examined and discussed. An improvement of 1.62mA/cm<SUP>2</SUP> in the short-circuit current density (J<SUB>sc</SUB>) is attributed to the higher band gap of p-type silicon oxide layers. In addition, an increase in open-circuit voltage (V<SUB>oc</SUB>) by 150mV and fill factor (FF) by 6.93% is ascribed to significantly improved front TCO/p-layer interface contact.

Attenuation of oxidative neuronal cell death by coffee phenolic phytochemicals

Cho, E.S.,Jang, Y.J.,Hwang, M.K.,Kang, N.J.,Lee, K.W.,Lee, H.J. Elsevier 2009 Mutation research Vol.661 No.1

Neurodegenerative disorders such as Alzheimer's disease (AD) are strongly associated with oxidative stress, which is induced by reactive oxygen species (ROS) including hydrogen peroxide (H<SUB>2</SUB>O<SUB>2</SUB>). Recent studies suggest that moderate coffee consumption may reduce the risk of neurodegenerative diseases such as AD, but the molecular mechanisms underlying this effect remain to be clarified. In this study, we investigated the protective effects of chlorogenic acid (5-O-caffeoylquinic acid; CGA), a major phenolic phytochemical found in instant decaffeinated coffee (IDC), and IDC against oxidative PC12 neuronal cell death. IDC (1 and 5μg/ml) or CGA (1 and 5μM) attenuated H<SUB>2</SUB>O<SUB>2</SUB>-induced PC12 cell death. H<SUB>2</SUB>O<SUB>2</SUB>-induced nuclear condensation and DNA fragmentation were strongly inhibited by pretreatment with IDC or CGA. Pretreatment with IDC or CGA also inhibited the H<SUB>2</SUB>O<SUB>2</SUB>-induced cleavage of poly(ADP-ribose) polymerase (PARP), and downregulation of Bcl-X<SUB>L</SUB> and caspase-3. The accumulation of intracellular ROS in H<SUB>2</SUB>O<SUB>2</SUB>-treated PC12 cells was dose-dependently diminished by IDC or CGA. The activation of c-Jun N-terminal protein kinase (JNK) and p38 mitogen-activated protein kinase (MAPK) by H<SUB>2</SUB>O<SUB>2</SUB> in PC12 cells was also inhibited by IDC or CGA. Collectively, these results indicate that IDC and CGA protect PC12 cells from H<SUB>2</SUB>O<SUB>2</SUB>-induced apoptosis by blocking the accumulation of intracellular ROS and the activation of MAPKs.