Evaluation of the zoonotic potential of a novel reassortant H1N2 swine influenza virus with gene constellation derived from multiple viral sources

Lee, J.H.,Pascua, P.N.Q.,Decano, A.G.,Kim, S.M.,Park, S.J.,Kwon, H.I.,Kim, E.H.,Kim, Y.I.,Kim, H.,Kim, S.Y.,Song, M.S.,Jang, H.K.,Park, B.K.,Choi, Y.K. Elsevier Science 2015 INFECTION GENETICS AND EVOLUTION Vol.34 No.-

In 2011-2012, contemporary North American-like H3N2 swine influenza viruses (SIVs) possessing the 2009 pandemic H1N1 matrix gene (H3N2pM-like virus) were detected in domestic pigs of South Korea where H1N2 SIV strains are endemic. More recently, we isolated novel reassortant H1N2 SIVs bearing the Eurasian avian-like swine H1-like hemagglutinin and Korean swine H1N2-like neuraminidase in the internal gene backbone of the H3N2pM-like virus. In the present study, we clearly provide evidence on the genetic origins of the novel H1N2 SIVs virus through genetic and phylogenetic analyses. In vitro studies demonstrated that, in comparison with a pre-existing 2012 Korean H1N2 SIV [A/swine/Korea/CY03-1½012 (CY03-1½012)], the 2013 novel reassortant H1N2 isolate [A/swine/Korea/CY0423/2013 (CY0423-12/2013)] replicated more efficiently in differentiated primary human bronchial epithelial cells. The CY0423-12/2013 virus induced higher viral titers than the CY03-1½012 virus in the lungs and nasal turbinates of infected mice and nasal wash samples of ferrets. Moreover, the 2013 H1N2 reassortant, but not the intact 2012 H1N2 virus, was transmissible to naive contact ferrets via respiratory-droplets. Noting that the viral precursors have the ability to infect humans, our findings highlight the potential threat of a novel reassortant H1N2 SIV to public health and underscore the need to further strengthen influenza surveillance strategies worldwide, including swine populations.

Enhanced H₂ fermentation of organic waste by CO₂ sparging

Kim, D.H.,Shin, H.S.,Kim, S.H. Pergamon Press ; Elsevier Science Ltd 2012 International journal of hydrogen energy Vol.37 No.20

This study aimed to improve the productivity of dark fermentative hydrogen production from organic waste. An anaerobic sequencing batch reactor was used for hydrogen fermentation and it was fed with food waste (VS 4.4 +/- 0.2% containing 27 g carbohydrate-COD/L) at various CO<SUB>2</SUB> sparging rates (40-120 L/L/d), hydraulic retention times (HRTs; 18-42 h), and solid retention times (SRTs; 18-160 h). CO<SUB>2</SUB> sparging increased the H<SUB>2</SUB> productivity by 5-36% at all the examined conditions, confirming the benefit of the replacement of headspace gas by CO<SUB>2</SUB>. The maximum H<SUB>2</SUB> production was obtained by CO<SUB>2</SUB> sparging at 80 L/L/d, resulting in the H<SUB>2</SUB> productivity of 3.18 L H<SUB>2</SUB>/L/d and the H<SUB>2</SUB> yield of 97.3 mL H<SUB>2</SUB>/g VS<SUB>added</SUB>. Increase of n-butyrate and isopropanol yields were concurrent with the enhanced H<SUB>2</SUB> yield by CO<SUB>2</SUB> sparging. Acidogenic efficiency, the sum of H<SUB>2</SUB>, organic acid, and alcohol, in the CO<SUB>2</SUB>-sparged reactor ranged from 47.9 to 56.0%, which was comparable to conventional acidogenesis. Thermodynamic analysis confirmed that both CO<SUB>2</SUB> sparging and CO<SUB>2</SUB> removal were beneficial for H<SUB>2</SUB>-producing reactions, but CO<SUB>2</SUB> sparing has more profound effect than CO<SUB>2</SUB> removal on inhibiting H<SUB>2</SUB>-consuming reactions.

Oxygen-dependent enhancement of hydrogen production by engineering bacterial hemoglobin in Escherichia coli

Jo, B.H.,Kim, J.Y.H.,Seo, J.H.,Cha, H.J. Pergamon Press ; Elsevier Science Ltd 2014 International journal of hydrogen energy Vol.39 No.20

H<SUB>2</SUB> production under aerobic conditions has been proposed as an alternative method to overcome the fundamentally low yield of H<SUB>2</SUB> production by fermentative bacteria by maximizing the number of electrons that are available for H<SUB>2</SUB>. Here, we engineered Vitreoscilla hemoglobin (VHb) in Escherichia coli to study the effects of this versatile oxygen (O<SUB>2</SUB>)-binding protein on oxic H<SUB>2</SUB> production in a closed batch system that was supplemented with glucose. The H<SUB>2</SUB> yields that were obtained with the VHb-expressing E. coli were greatly enhanced in comparison to the negative control cells in culture that started with high O<SUB>2</SUB> tensions. The formate hydrogen lyase (FHL) activity of oxically cultured, VHb-expressing cells was also much higher than that of the negative control cells. Through inhibitor studies and time-course experiments, VHb was shown to contribute to the improved H<SUB>2</SUB> yield primarily by increasing the efficiency of cellular metabolism during the aerobic phase before the onset of H<SUB>2</SUB> production and not by working as an O<SUB>2</SUB>-scavenger during H<SUB>2</SUB> production. This new approach allowed more substrate to remain to be further utilized for the production of more H<SUB>2</SUB> from limited resources. We expect that VHb can be successfully engineered in potential aerobic H<SUB>2</SUB>-producing microbial systems to enhance the overall H<SUB>2</SUB> production yield. In addition, the remarkably high FHL activity of oxically grown, VHb-expressing cells may make this engineered strain an attractive whole-cell biocatalyst for converting formate to H<SUB>2</SUB>.

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>

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.

Synthesis, crystal structures, and deprotonation of cis- and trans-octahedral nickel(II) complexes with a 14-membered tetraaza macrocycle bearing two N-phenacyl pendant arms

Kim, H.,Kang, S.G.,Kwak, C.H. Elsevier Sequoia [etc.] 2012 Inorganica chimica acta Vol.387 No.-

The di-N-functionalized macrocycle 2,13-bis(2-phenacyl)-5,16-dimethyl-2,6,13,17-tetraazatricyclo[16.4.0.<SUP>1.18</SUP>0<SUP>7.12</SUP>]docosane (H<SUB>2</SUB>L<SUP>2</SUP>) bearing two N-CH<SUB>2</SUB>COC<SUB>6</SUB>H<SUB>5</SUB> groups has been prepared by the reaction of 3,14-dimethyl-2,6,13,17-tetraazatricyclo[16.4.0.0<SUP>7.12</SUP>]docosane (L<SUP>1</SUP>) with phenacyl bromide. Interestingly, H<SUB>2</SUB>L<SUP>2</SUP> reacts with Ni<SUP>2+</SUP> ion to form two geometric isomers, trans-[Ni(H<SUB>2</SUB>L<SUP>2</SUP>)]<SUP>2+</SUP> and cis-[Ni(H<SUB>2</SUB>L<SUP>2</SUP>)]<SUP>2+</SUP>. The axial Ni-O (N-CH<SUB>2</SUB>COC<SUB>6</SUB>H<SUB>5</SUB> group) bond distance (2.080(2)A) of trans-[Ni(H<SUB>2</SUB>L<SUP>2</SUP>)](ClO<SUB>4</SUB>)<SUB>2</SUB>.2DMSO is shorter than the in-plane Ni-N distances (2.096(2) and 2.100(2)A). However, the Ni-O distances (2.105(2) and 2.124(2)A) of cis-[Ni(H<SUB>2</SUB>L<SUP>2</SUP>)](ClO<SUB>4</SUB>)<SUB>2</SUB>.H<SUB>2</SUB>O are considerably longer than the Ni-N distances (2.053(2)-2.086(2)A). Each N-CH<SUB>2</SUB>COC<SUB>6</SUB>H<SUB>5</SUB> group of trans-[Ni(H<SUB>2</SUB>L<SUP>2</SUP>)]<SUP>2+</SUP> and cis-[Ni(H<SUB>2</SUB>L<SUP>2</SUP>)]<SUP>2+</SUP> exists as its keto form in the solid state and in various solvents. Two N-CH<SUB>2</SUB>COC<SUB>6</SUB>H<SUB>5</SUB> groups of trans-[Ni(H<SUB>2</SUB>L<SUP>2</SUP>)]<SUP>2+</SUP> are readily deprotonated in basic aqueous solutions, producing the enolate form trans-[Ni(L<SUP>2</SUP>)]. On the other hand, cis-[Ni(H<SUB>2</SUB>L<SUP>2</SUP>)]<SUP>2+</SUP> undergoes deprotonation to yield cis-[Ni(HL<SUP>2</SUP>)]<SUP>+</SUP>, in which one N-CH<SUB>2</SUB>COC<SUB>6</SUB>H<SUB>5</SUB> group is not deprotonated, under similar conditions.

Cold stress causes rapid but differential changes in properties of plasma membrane H⁺-ATPase of camelina and rapeseed

Kim, H.S.,Oh, J.M.,Luan, S.,Carlson, J.E.,Ahn, S.J. G. Fischer 2013 Journal of plant physiology Vol.170 No.9

Camelina (Camelina sativa) and rapeseed (Brassica napus) are well-established oil-seed crops with great promise also for biofuels. Both are cold-tolerant, and camelina is regarded to be especially appropriate for production on marginal lands. We examined physiological and biochemical alterations in both species during cold stress treatment for 3 days and subsequent recovery at the temperature of 25<SUP>o</SUP>C for 0, 0.25, 0.5, 1, 2, 6, and 24h, with particular emphasis on the post-translational regulation of the plasma membrane (PM) H<SUP>+</SUP>-ATPase (EC3.6.3.14). The activity and translation of the PM H<SUP>+</SUP>-ATPase, as well as 14-3-3 proteins, increased after 3 days of cold stress in both species but recovery under normal conditions proceeded differently. The increase in H<SUP>+</SUP>-ATPase activity was the most dramatic in camelina roots after recovery for 2h at 25<SUP>o</SUP>C, followed by decay to background levels within 24h. In rapeseed, the change in H<SUP>+</SUP>-ATPase activity during the recovery period was less pronounced. Furthermore, H<SUP>+</SUP>-pumping increased in both species after 15min recovery, but to twice the level in camelina roots compared to rapeseed. Protein gel blot analysis with phospho-threonine anti-bodies showed that an increase in phosphorylation levels paralleled the increase in H<SUP>+</SUP>-transport rate. Thus our results suggest that cold stress and recovery in camelina and rapeseed are associated with PM H<SUP>+</SUP>-fluxes that may be regulated by specific translational and post-translational modifications.

Effect of substrate concentration on continuous Photo-fermentative hydrogen production from lactate using Rhodobacter sphaeroides

Kim, D.H.,Son, H.,Kim, M.S. Pergamon Press ; Elsevier Science Ltd 2012 INTERNATIONAL JOURNAL OF HYDROGEN ENERGY - Vol.37 No.20

The information on continuous operation and the use of actual waste as a feedstock are essential for the practical application of photo-fermentative H<SUB>2</SUB> production. For the first 200 days, continuous H<SUB>2</SUB> production from lactate was attempted using purple non-sulfur (PNS) bacteria, Rhodobacter sphaeroides KD131, under an illumination of 110 W/m<SUP>2</SUP>. During the continuous operation, 30% of the fermenter volume was replaced by fresh feedstock once a day, and substrate concentration was gradually increased from 5 mM to 30 mM. H<SUB>2</SUB> production was negligible at 5 mM, which was ascribed to the fact that the electrons contained in lactate were mostly consumed for cell growth and soluble microbial products (SMPs) production. As lactate concentration increased, H<SUB>2</SUB> production gradually increased and reached a maximum at 20 mM, showing a substrate conversion efficiency (SCE) of 38%, a H<SUB>2</SUB> yield of 2.3 mol H<SUB>2</SUB>/mol lactate<SUB>added</SUB>, and a H<SUB>2</SUB> production rate of 309 mL H<SUB>2</SUB>/L-fermenter/d. Further increases of lactate concentration resulted in a drop of H<SUB>2</SUB> production (<1.0 mol H<SUB>2</SUB>/mol lactate<SUB>added</SUB>). When the feedstock was changed to actual waste obtained from a 1-day lactate fermentation of food waste, stable H<SUB>2</SUB> production was maintained, but showed a decreased SCE of 24%. It was speculated that the low performance was due to the fact that actual waste contained not only pure lactate but also other organic compounds that could not be utilized by PNS bacteria. In addition, compared to feeding with pure lactate, the electron consumption to the cell growth was higher in feeding with actual waste, which led to the lower performance.

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>

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.