Expression and N-glycan analysis of human 90K glycoprotein in Drosophila S2 cells

Kim, K.R.,Kim, Y.K.,Cheong, H.,Kim, J.Y.H.,Cha, H.J. IPC Science and Technology Press ; Elsevier Scienc 2013 Enzyme and microbial technology Vol.53 No.3

Human 90K (h90K; Mac-2-binding protein) glycoprotein is a potential pharmaceutical due to its inhibitory activity against cancer metastasis and expansion. Here, h90K glycoprotein was produced in insect Drosophila S2 cell system, and its N-glycan pattern was analyzed. A plasmid encoding h90K gene, fused with a hexahistidine tag under the control of Drosophila metallotionein promoter, was stably transfected into S2 cells. After copper sulfate induction, transfected S2 cells secreted recombinant h90K at a good expression level of 28mg/L in a 150-mL spinner flask culture. The purified recombinant h90K showed an apparent molecular weight of ~78kDa which was much smaller than that (~97kDa) of the natural h90K. Because de-N-glycosylated h90K appeared at ~60kDa protein band, it was suggested that the recombinant h90K from S2 cells has small N-glycans with about half the molecular weight (~18kDa) of N-glycans of the natural h90K. Through detail analyses using high-performance liquid chromatography and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, the S2-derived recombinant h90K was confirmed that it has simple paucimannosidic structures containing two or three mannose residues with core fucose as the major (~79%) N-glycans.

Induction of Cell Death in Human Macrophages by a Highly Virulent Korean Isolate of <i>Mycobacterium tuberculosis</i> and the Virulent Strain H37Rv

Sohn, H.,Lee, K.-S.,Kim, S.-Y.,Shin, D.-M.,Shin, S.-J.,Jo, E.-K.,Park, J.-K.,Kim, H.-J. Blackwell Publishing Ltd 2009 Scandinavian journal of immunology Vol.69 No.1

<P>Abstract</P><P>Recent studies have suggested that virulent strains of <I>Mycobacterium tuberculosis</I> induce apoptosis in macrophages less often than do attenuated strains. K-strain, which belongs to the Beijing family, is the most frequently isolated clinical strain of <I>M. tuberculosis</I> in Korea. In this study, we investigated the differential induction of cell death in human monocytic THP-1 cells by K-strain and H37Rv, a virulent but laboratory-adapted strain of <I>M. tuberculosis</I>. Although no significant difference in growth rate was observed between the cells exposed to K-strain and those exposed to H37Rv, the levels of protective cytokines such as tumour necrosis factor (TNF)-&agr;, interleukin (IL)-6 and IL-12p40 were lower in K-strain-infected cells than in H37Rv-infected cells. Cell viability assays showed that both K-strain and H37Rv, but not heat- or streptomycin-killed bacteria, induced THP-1 cell death in a TNF-independent manner. In contrast, double staining with fluorochrome-labelled inhibitors of caspase and propidium iodide and lactate dehydrogenase release assays revealed that K-strain induced significantly higher levels of necrotic cell death, rather than apoptosis, in THP-1 cells than did H37Rv. Anti-apoptotic <I>Bcl-2</I>, <I>Mcl-1</I>, <I>Bfl-1</I> and <I>Bcl-xL</I> in the cells were significantly upregulated following infection with K-strain compared with H37Rv, whereas <I>Bax</I> was slightly upregulated in response to infection with both H37Rv and K-strain. These results suggest that the highly virulent K-strain keeps cellular apoptosis as a host defense mechanism to a minimum and induces necrosis in macrophages.</P>

H3K27 methylation and H3S28 phosphorylation-dependent transcriptional regulation by INHAT subunit SET/TAF-Iβ

Kim, J.Y.,Kim, K.B.,Son, H.J.,Chae, Y.C.,Oh, S.T.,Kim, D.W.,Pak, J.H.,Seo, S.B. North-Holland Pub ; Elsevier Science Ltd 2012 FEBS letters Vol.586 No.19

Significant progress has been made in understanding the relationship between histone modifications and 'reader' molecules and their effects on transcriptional regulation. A previously identified INHAT complex subunit, SET/TAF-Iβ, binds to histones and inhibits histone acetylation. To investigate the binding specificities of SET/TAF-Iβ to various histone modifications, we employed modified histone tail peptide array analyses. SET/TAF-Iβ strongly recognized PRC2-mediated H3K27me½/3; however, the bindings were completely disrupted by H3S28 phosphorylation. We have demonstrated that SET/TAF-Iβ is sequentially recruited to the target gene promoter ATF3 after the PRC2 complex via H3K27me recognition and may offer additive effects in the repression of the target gene.

Genetic and phylogenetic characterizations of a novel genotype of highly pathogenic avian influenza (HPAI) H5N8 viruses in 2016/2017 in South Korea

Kim, Y.I.,Park, S.J.,Kwon, H.I.,Kim, E.H.,Si, Y.J.,Jeong, J.H.,Lee, I.W.,Nguyen, H.D.,Kwon, J.J.,Choi, W.S.,Song, M.S.,Kim, C.J.,Choi, Y.K. Elsevier Science 2017 INFECTION GENETICS AND EVOLUTION Vol.53 No.-

<P>During the outbreaks of highly pathogenic avian influenza (HPAI) H5N6 viruses in 2016 in South Korea, novel H5N8 viruses were also isolated from migratory birds. Phylogenetic analysis revealed that the HA gene of these H5N8 viruses belonged to clade 2.3.4.4, similarly to recent H5Nx viruses, and originated from A/Brk/Korea/Gochang1/14(H5N8), a minor lineage of H5N8 that appeared in 2014 and then disappeared. At least four reassortment events occurred with different subtypes (H5N8, H7N7, H3N8 and H10N7) and a chicken challenge study revealed that they were classified as HPAI viruses according to OIE criteria. (C) 2017 Elsevier B.V. All rights reserved.</P>

Relationship between Stress Gene Polymorphisms and Litter Size by AI in Pigs

H. J. Jin,I. C. Kim,M. S. Wee,S. H. Yeon,C. D. Kim,S. S. Lee,C. Y. Cho,S. R. Cho,D. S. Son,C. K. Park,Z. D. Li 사단법인 한국동물생명공학회 2007 한국동물생명공학회지 Vol.22 No.4

This study was performed to investigate the relationship between PSS-HSP70 gene polymorphism and artificial insemination (AI) reproductivity in the pigs. The RFLP polymorphism of PSS and the SSCP polymorphisms of HSP70 K1, K3 and K4 PCR product were detected different patterns. In the experiment for AI of fresh semen, spring and fall season showed higher litter size born of 10.89 head than 10.47 head of summer season. Landrace was showed higher litter size of 9.96 head than that of Duroc and Yorkshire(p<0.05). Stress relating PSS and HSP70 polymorphism of PSS-Normal, HSP70 K1-BB, K3-AB, K4-AA showd a highest litter size born of 10.97 head and litter size born alive of 10.69 head than that of the other polymorphisms(p<0.05). In the experiment for AI of frozen semen, effects of season and pig breeds were not showed for litter size born. The stress relating polymorphism of PSS-Carrier, HSP70 K1-BB, K3-BB, K4-AB showed highest litter size born of 11.29 head and litter size born alive of 10.82 head and PSS-Normal, HSP70 K1-BB, K3-AB, K4-AA showed the lowest litter size born of 8.48 head and litter size born alive of 7.33 head than that of the other polymorphisms(p<0.05). These results suggest that AI litter size born for the stress of forzen thawed semen may be affected by PSS and HSP70 polymorphism in pigs.

Genetic diversity and pathogenic potential of low pathogenic H7 avian influenza viruses isolated from wild migratory birds in Korea

Kim, Y.I.,Kim, S.W.,Si, Y.J.,Kwon, H.I.,Park, S.J.,Kim, E.H.,Kim, S.m.,Lee, I.W.,Song, M.S.,Choi, Y.K. Elsevier Science 2016 Infection, genetics and evolution Vol.45 No.-

To detect the circulation of H7 avian influenza viruses, we characterized H7 viruses found in migratory birds and live poultry markets of South Korea from 2005 to 2014. Phylogenic analysis revealed that while all viruses clustered into the Eurasian-lineage of H7 avian viruses, at least 12 distinct genotypes were represented. Most H7 viruses contained at least one gene segment from the highly-pathogenic A/Sck/Hong Kong/YU100/02(H5N1)-like avian virus, and they could be separated into at least two antigenic groups. Although we did not detect genetically identical strains, HI assay demonstrated close cross-reactivity of some isolates with the H7N9 viruses from China. Animal studies revealed that most of the genotypes could replicate in the lungs of mice and chickens without prior adaptation and some, particularly H7N4 and H7N7 subtypes, induced mortality in mice. These results reinforce growing pandemic concerns regarding recent H7 viruses and emphasize the importance of continued surveillance of avian influenza viruses in the wild.

Ex situ catalytic upgrading of lignocellulosic biomass components over vanadium contained H-MCM-41 catalysts

Kim, B.S.,Jeong, C.S.,Kim, J.M.,Park, S.B.,Park, S.H.,Jeon, J.K.,Jung, S.C.,Kim, S.C.,Park, Y.K. Elsevier Science Publishers 2016 CATALYSIS TODAY - Vol.265 No.-

<P>H-V-MCM-41 catalysts containing 5, 10, and 30 wt% of vanadium were synthesized and applied to the ex situ catalytic pyrolysis (CP) of three polymeric components of lignocellulosic biomass for the first time. Characterization of the catalysts was performed using N-2 adsorption-desorption, XRD, FT-IR, and NH3-TPD. The results of XRD analysis showed that 5 wt% and 10 wt% H-V-MCM-41 catalysts maintained the mesoporous structure, whereas the mesoporous structure was destroyed in 30 wt% H-V-MCM-41 with considerable amount of small V2O5 crystalline outside the framework. NH3-TPD showed that H-V-MCM-41 has mostly weak acid sites and that 10 wt% H-V-MCM-41 had the largest quantity of acid sites due to framework vanadium. In the case of CP of cellulose using Py-GC/MS, 10 wt% H-V-MCM-41 showed the highest catalytic activity for the production of valuable furanic compounds such as furfural because of the enhanced deoxygenation over the acid sites formed on framework vanadium. In the case of CP of xylan as well, 10 wt% H-V-MCM-41 led to the largest yield of mono-aromatics. The production of acetic acid was also promoted by H-V-MCM-41 catalysts. The CP of lignin over H-V-MCM-41 catalysts promoted substantially the production of important feedstock chemicals for the petrochemical industry: phenolics and mono-aromatics. (C) 2015 Elsevier B.V. All rights reserved.</P>

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.

Aquaporin 2-labeled cells differentiate to intercalated cells in response to potassium depletion

Kim, W. Y.,Nam, S. A.,Choi, A.,Kim, Y. M.,Park, S. H.,Kim, Y. K.,Kim, J. Springer Science + Business Media 2016 Histochemistry and cell biology Vol.145 No.1

<P>The mammalian renal collecting duct consists of principal cells (PCs) and intercalated cells (ICs). Both PCs and ICs are involved in potassium (K+) homeostasis, PCs through their role in K+ secretion and ICs through their ability to facilitate K+ resorption. We previously hypothesized that PCs may differentiate into ICs upon K+ depletion. However, no direct evidence has yet been obtained to conclusively demonstrate that PCs differentiate into ICs in response to K+ depletion. Here, we present direct evidence for the differentiation of PCs into ICs by cell lineage tracing using aquaporin 2 (AQP2)-Cre mice and R26R-EYFP transgenic mice. In control mice, AQP2-EYFP+ cells exhibited mainly a PC phenotype (AQP2-positive/H+-ATPase-negative). Interestingly, some AQP2-EYFP+ cells exhibited an IC phenotype (H+-ATPase-positive/AQP2-negative); these cells accounted for 1.7 %. After K+ depletion, the proportion of AQP2-EYFP+ cells with an IC phenotype was increased to 4.1 %. Furthermore, some AQP2-EYFP+ cells exhibited a 'null cell' phenotype (AQP2-negative/H+-ATPase-negative) after K+ depletion. Collectively, our data demonstrate that AQP2-labeled cells can differentiate into ICs, as well as null cells, in response to K+ depletion. This finding indicates that some of AQP2-labeled cells possess properties of progenitor cells and that they can differentiate into ICs in the adult mouse kidney.</P>

WO₃ nanofibers functionalized by protein-templated RuO₂ nanoparticles as highly sensitive exhaled breath gas sensing layers

Kim, K.H.,Kim, S.J.,Cho, H.J.,Kim, N.H.,Jang, J.S.,Choi, S.J.,Kim, I.D. Elsevier Sequoia 2017 Sensors and actuators. B, Chemical Vol.241 No.-

<P>In this work, a novel catalytic synthesis and functionalization method using apoferritin is used to fabricate RuO2 nanoparticles (NPs) loaded WO3 nanofibers (NFs) for potential diagnosis of diabetes. Catalytic ruthenium (Ru) NPs with very small average diameters of 1.8 +/- 0.9 nm were synthesized using apoferritin which is a hollow protein cage, and were easily functionalized on WO3 NFs by introducing electrospinning solution with W precursor and polyvinylpyrrolidone (PVP). As-spun Ru NPs-loaded W precursor/PVP composite NFs were calcined at 600 degrees C for 1 h in air atmosphere to achieve RuO2-functionalized WO3 NFs. The small size and uniform distribution of catalytic RuO2 NPs were well maintained due to hollow nature of apoferritin cages after calcination. The chemo-resistive sensors using RuO2-functionalized WO3 NFs showed significantly enhanced acetone (CH3COCH3) sensing response (R-air/R-gas = 78.61-5 ppm), which was 7.4 times higher than the response (R-air/R-gas =10.61-5 ppm) of pristine WO3 NFs at highly humid atmosphere (95% RH). In addition, the RuO2-functionalized WO3 NFs showed outstanding selectivity toward acetone gas in comparison with other gases such as hydrogen sulfide (H2S), toluene (C6H5CH3), ethanol (C2H5OH), pentane (C5H12), ammonia (NH3), hydrogen (H-2), and water vapor (H2O) at 5 ppm. These results represent potential feasibility for the detection of acetone in exhaled breath for diagnosis of diabetes. (C) 2016 Elsevier B.V. All rights reserved.</P>