Escherichia coli-based expression of functional novel DNA-binding histone H1 from Carassius auratus

Wei, Q.,Jung, H.J.,Hwang, D.S.,Hwang, B.H.,Gim, Y.,Cha, H.J. IPC Science and Technology Press ; Elsevier Scienc 2007 Enzyme and microbial technology Vol.40 No.6

Histones are DNA-binding proteins that assist in DNA packaging and protection. Here, we, for the first time, cloned a novel histone H1 cDNA (638bp) from the goldfish, Carassius auratus. Sequencing revealed that this histone H1 shared 68.1% amino acid identity and 73.9% similarity with that from the rainbow trout, Salmo gairdneri. We successfully expressed a full-length recombinant version (∼20kDa) and a truncated C-terminal fragment (∼6kDa; 61 amino acids) of this histone H1 as a partially soluble form using a maltose binding protein (MBP) fusion strategy in an Escherichia coli expression system. Our results revealed that both these recombinant histone H1 versions had DNA binding and protection functions, and MBP fusion system was an effective way to produce biological functional recombinant histone proteins in E. coli. This novel recombinant histone H1 protein and/or its C-terminal peptide could be used as potential mediators for efficacious gene delivery.

Effect of the Length of Feed Withdrawal on Weight Loss, Yield and Meat Color of Broiler

Kim, D.H.,Yoo, Y.M.,Kim, S.H.,Jang, B.G.,Park, B.Y.,Cho, S.H.,Seong, P.N.,Hah, K.H.,Lee, J.M.,Kim, Y.K.,Hwang, I.H. Asian Australasian Association of Animal Productio 2007 Animal Bioscience Vol.20 No.1

The current study was conducted to determine the optimum length of feed withdrawal for pre-harvest broilers. A total of three hundred broilers were sampled from an industrial population, and 30 chicks for each weight group (e.g., 1.5 and 2.5 kg) were randomly assigned to feed withdrawal treatments for 0, 3, 6, 9 and 12 h. Weight loss, yield, muscle pH, objective meat color and weights of gastro intestinal contents, crop, gizzard, provenriculus, small intestine, caecum, and rectum were determined. Live weight loss was significantly (p<0.05) increased as length of feed withdrawal extended. A significant (p<0.05) carcass yield for both 1.5 and 2.5 kg groups coincided after 9 and 6 h feed withdrawal, respectively. Net weights of intestinal contents for crop and gizzard were significantly (p<0.05) reduced by 6 h, and the reduction for proventriculus and small intestine occurred from 3 h. A noticeable effect of feed withdrawal on pH for breast muscle at 3 h postmortem occurred only when chicks were fasted for 3 h of which pH (6.05) was significantly (p<0.05) higher than that for other groups including the control (5.74). There was a linear tendency of higher lightness (Hunter L* value) numerically for chicks fasted for longer periods. The highest coefficient of determinations of regression models to estimate weight loss as a function of fasting period and body weights were achieved, when the models included both linear and quadratic terms for fasting period, and linear term for both 1.5 ($R^2=0.76$) and 2.5 kg ($R^2=0.78$) body weight groups. Given the practical aspect, approximately 1.5 kg of body weight is dominant, weight loss could be predicted by the following function; live weight $loss=26.6-0.28{\times}(fasting period)^2+12.34{\times}pasting\;period-0.012{\times}body\;weight$, $R^2=0.76$. Current data implied that the optimum fasting time for pre-slaughter chicks varied depending on slaughter weight; 6 and 9-h fasting were recommendable for 2.5 and 1.5 kg chicks, with little effect on objective meat color.

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>

Kinetic study on coal-biomass mixed char co-gasification with H₂O in the presence of H₂

Massoudi Farid, M.,Jeong, H.J.,Hwang, J. Butterworths [etc.] ; Elsevier Science Ltd 2016 Fuel Vol.181 No.-

The coal-biomass mixed char co-gasification with H<SUB>2</SUB>O in the presence of H<SUB>2</SUB> was investigated under atmospheric conditions using a tube furnace and gas analyzer. Various partial pressures of H<SUB>2</SUB>O and H<SUB>2</SUB> were tested within a temperature range of 825-900<SUP>o</SUP>C. Bituminous coal, pineapple sawdust, and a mixture of these two having a mass ratio of 1:1 were used to prepare chars. Random pore model was utilized to explain the experimental results. The results demonstrated that the inhibition effect of H<SUB>2</SUB> on char-H<SUB>2</SUB>O gasification can be described using the Langmuir-Hinshelwood (L-H) equation. The inhibition effect on the mixture sample was more intense than on the pure samples. The L-H equation kinetic parameters were acquired and expressed in an Arrhenius equation form to determine the pre-exponential factor and activation energy.

오픈소스 LinuxCNC를 사용한 듀얼서보모터 H-TYPE 공작기계 Test-bed 구축

장현수(H. S. Jang),강유수(Y. S. Kang),황준호(J. H. Hwang),서영호(Y. H. Seo),최인휴(I. H. Choi),김병희(B. H. Kim) 한국생산제조학회 2021 한국생산제조시스템학회 학술발표대회 논문집 Vol.2021 No.7

Dipeptidyl petidase-IV inhibitor (gemigliptin) inhibits tunicamycin-induced endoplasmic reticulum stress, apoptosis and inflammation in H9c2 cardiomyocytes

Hwang, H.J.,Jung, T.W.,Ryu, J.Y.,Hong, H.C.,Choi, H.Y.,Seo, J.A.,Kim, S.G.,Kim, N.H.,Choi, K.M.,Choi, D.S.,Baik, S.H.,Yoo, H.J. North-Holland 2014 Molecular and cellular endocrinology Vol.392 No.1

The direct effects of dipeptidyl peptidase-IV (DPP-IV) inhibitors on endoplasmic reticulum (ER) stress-induced apoptosis and inflammation in cardiomyocytes have not been elucidated. H9c2 cell viability, which was reduced by tunicamycin, was increased after DPP-IV inhibitor gemigliptin treatment. Gemigliptin significantly decreased the tunicamycin-mediated increase in glucose regulated protein 78 (GRP78) expression and ER stress-mediated signaling molecules such as protein kinase RNA-like endoplasmic reticulum kinase (PERK)/C-EBP homologous protein (CHOP) and inositol-requiring enzyme 1α (IRE1α)/c-Jun N-terminal kinase (JNK)-p38. Furthermore, gemigliptin effectively induced Akt phosphorylation in a dose-dependent manner. Using flow cytometry and Hoechst staining, we showed that treatment with Akt inhibitor significantly blocked the anti-apoptotic effects mediated by gemigliptin. The reduction in tunicamycin-induced GRP78 level and PERK/CHOP pathway activity by gemigliptin was reversed after treatment with Akt inhibitor. In conclusion, gemigliptin effectively inhibited ER stress-induced apoptosis and inflammation in cardiomyocytes via Akt/PERK/CHOP and IRE1α/JNK-p38 pathways, suggesting its direct protective role in cardiovascular diseases.

Reclassification of Halomonas caseinilytica Wu et al. 2008 as a later synonym of Halomonas sinaiensis Romano et al. 2007, and emendation of the species description

Hwang, C. Y.,Lee, I.,Yoon, S. J.,Lee, Y. M.,Lee, W. S.,Lee, H. K. Springer Science + Business Media 2016 Antonie van Leeuwenhoek Vol.109 No.10

<P>The taxonomic relationship between Halomonas sinaiensis DSM 18067(T) and Halomonas caseinilytica JCM 14802(T) has not been established, despite the high similarity (99.6 %) of their 16S rRNA gene sequences. To clarify their taxonomic positions, a polyphasic approach was applied to both type strains. Genomic relatedness analyses between H. sinaiensis DSM 18067(T) and H. caseinilytica JCM 14802(T) resulted in an average nucleotide identity of 99.5 % and an estimated DNA-DNA hybridization of 96.1 % by the genome-to-genome distance calculator, indicating that they belong to a single species. Phenotypic and chemotaxonomic characteristics showed no pronounced differences between the two type strains. Based on the results of this polyphasic study, it is proposed that H. caseinilytica JCM 14802(T) is a later heterotypic synonym of H. sinaiensis DSM 18067(T). An emended description for the species H. sinaiensis is given.</P>

First molecular cloning and gene expression analysis of a teleost CD200 (OX-2) glycoprotein from rock bream, Oplegnathus fasciatus

Hwang, S.D.,Kim, J.W.,Kim, M.C.,Kim, D.H.,Park, C.I. Academic Press 2013 Fish & shellfish immunology Vol.34 No.1

CD200 plays an important role in delivering an immunoregulatory signal to the immune system through interaction with its receptor. However, CD200 has not been characterized and its function in teleosts is unknown. In this study, the rock bream (Oplegnathus fasciatus) CD200 gene (RbCD200) was cloned and its expression profile was analyzed after infection with Edwardsiella tarda, Streptococcus iniae or red seabream iridovirus (RSIV). The coding region of RbCD200 cDNA was 855 bp, encoding 284 amino acid residues. The gene consisted of two extracellular Ig-like domains and a transmembrane domain. RbCD200 was highly expressed in the brain, erythrocytes, intestine and stomach of healthy rock bream. In the spleen, RbCD200 gene expression was down-regulated until 48 h after E. tarda exposure, except at 12 h RbCD200 gene expression was down-regulated then up-regulated at 12 h and 24 h after infection with S. iniae and RSIV, respectively. In the whole kidney, the RbCD200 gene was down-regulated in response to infection with E. tarda and S. iniae. However, RSIV infection increased RbCD200 gene expression in whole kidney until 48 h. These results suggest that RbCD200 is differentially expressed in the spleen and whole kidney after infection with different pathogens.

O-free polyacrylonitrile doping to improve the J_c(B) and H_c2 of MgB₂ wires

Hwang, S.M.,Sung, K.,Choi, J.H.,Kim, W.,Joo, J.,Lim, J.H.,Kim, C.J.,Park, Y.S.,Kim, D.H. North-Holland 2010 Physica. C, Superconductivity Vol.470 No.20

We selected polyacrylonitrile (PAN, -[C<SUB>3</SUB>H<SUB>3</SUB>N]-) as an O-free organic dopant and fabricated C-doped MgB<SUB>2</SUB> wires by in situ and powder-in-tube techniques. 0-5 wt.% PAN powders were uniformly mixed with B powder using a liquid mixing method. The precursor powders were mixed with Mg powder, filled into Fe tubes, and then drawn into wires. Sintering was performed at 900<SUP>o</SUP>C for 1h in a flowing Ar gas. The PAN doping decreased the critical temperature (T<SUB>c</SUB>) and a-axis lattice parameter, but significantly improved the critical current density (J<SUB>c</SUB>) in high fields, upper critical field (H<SUB>c2</SUB>), and irreversibility field (H<SUB>irr</SUB>) performances. These results are attributed to the replacement of B sites with C by the PAN doping. Furthermore, as expected, the MgO amount did not increase as the doping content increased. The J<SUB>c</SUB> of the PAN-doped MgB<SUB>2</SUB> wires was more than one order of magnitude higher than that of the undoped MgB<SUB>2</SUB> wire at 5K and 6.6T (1.46-3.82kA/cm<SUP>2</SUP> vs. 0.11kA/cm<SUP>2</SUP>).

Application of Gel-based Proteome Analysis Techniques to Studying Post-mortem Proteolysis in Meat

Hwang, I.H. Asian Australasian Association of Animal Productio 2004 Animal Bioscience Vol.17 No.9

This study was conducted to evaluate the possible application of 2 D-SDS-PAGE (2 DE)-based proteome analysis techniques to the assessment of extreme proteolysis in postmortem skeletal muscle. Eight Hanwoo longissimus muscles were incubated immediately after slaughter for 24 h at 5$^{\circ}C$, 15$^{\circ}C$ or 36$^{\circ}C$. Warner Bratzler (WB)-shear force and ultrastructural configuration were determined at 24 h, and rate of proteolysis to 24 h was determined by 1 D-SDS-PAGE (1 DE) and 2 DE. In addition, tentative protein identification was performed from peptide mass fingerprints of MALDI-ToF analysis of major protein groups on 2 DE profiles. The result showed that although ultrastructural configuration was similar between the 5$^{\circ}C$ and 36$^{\circ}C$ treatments, meat at 5$^{\circ}C$ had higher WBshear force (approximately 5 kg greater). A higher rate of protein degradation at 36$^{\circ}C$ was observed based on Troponin-T degradation, 1 DE, and 2 DE analysis. This indicates that proteolysis during the early postmortem period was a significant determinant of shear force at 24 h. Little difference in proteolysis between 5$^{\circ}C$ and 15$^{\circ}C$ treatments was found based on classic 1 DE profile assessment. Meanwhile, considerable differences in the 2 DE profiles between the two treatments were revealed, with substantially higher rate of proteolysis at 15$^{\circ}C$ compared to 5$^{\circ}C$. Nuclease treatment improved 2 DE profile resolution. 400 ${\mu}$g and 600 ${\mu}$g of sample loading appeared to be appropriate for 24 cm pH 3-10 and pH 5-7 IPG strips, respectively. Protein detection and quantification of the 5$^{\circ}C$, 15$^{\circ}C$ and 36$^{\circ}C$ 2 DE profiles revealed 78, 163 and 232 protein spots respectively that were differentially modified in terms of their electrophoretic properties between approximately pI 5.3-7.7 with the molecular weight range of approximately 71-12 kDa. The current results demonstrated that 2 DE was a superior tool to 1 DE for characterising proteolysis in postmortem skeletal muscle.