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.

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.

Antiviral phenolics from the leaves of Cleistocalyx operculatus

Ha, T.K.Q.,Dao, T.T.,Nguyen, N.H.,Kim, J.,Kim, E.,Cho, T.Oh.,Oh, W.K. Elsevier 2016 Fitoterapia Vol.110 No.-

<P>During the screening program for anti-influenza agents from medicinal plants, the ethanolic extract of Cleistocalyx operculatus leaves was found to exhibit potential neuraminidase (NA) inhibitory activity. Bioassay directed fractionation led to the isolation of two new acetophenones (1 and 2) and one new flavanone (3), along with six known compounds (4-9). The structures of all isolated compounds were elucidated using various spectroscopic methods and through comparison with the previous literature. Compounds 6 and 8 exhibited strong enzymatic inhibition on various neuraminidases from different influenza viruses, including H1N1, H9N2, novel H1N1, and oseltamivir-resistant novel H1N1 (H274Y mutation) expressed in HEM293 cells (IC50 values ranging from 5.07 +/- 0.94 mu M to 9.34 +/- 2.52 mu M, respectively). Kinetic experiments revealed the non-competitive inhibitory mode of both compounds 6 and 8. Furthermore, these flavonoids reduced the cytopathic effect of the H1N1 virus in MDCK cells. The present study suggests the potential of two flavonoids (6 and 8) as new lead compounds for the development of novel NA inhibitors in the future. (C) 2016 Elsevier B.V. All rights reserved.</P>

Effects of L-tryptophan, Fructan, and Casein on Reducing Ammonia, Hydrogen Sulfide, and Skatole in Fermented Swine Manure

Q.K. Sheng,Z.J. Yang,H.B. Zhao,X.L. Wang,J.F. Guo 아세아·태평양축산학회 2015 Animal Bioscience Vol.28 No.8

The effects of daily dietary Bacillus subtilis (Bs), and adding L-tryptophan, fructan, or casein to fecal fermentation broths were investigated as means to reduce the production of noxious gas during manure fermentation caused by ammonia, hydrogen sulfide (H2S), and 3-methylindole (skatole). Eighty swine (50.0±0.5 kg) were equally apportioned to an experimental group given Bs in daily feed, or a control group without Bs. After 6 weeks, fresh manure was collected from both groups for fermentation studies using a 3×3 orthogonal array, in which tryptophan, casein, and fructan were added at various concentrations. After fermentation, the ammonia, H2S, L-tryptophan, skatole, and microflora were measured. In both groups, L-tryptophan was the principle additive increasing skatole production, with significant correlation (r = 0.9992). L-tryptophan had no effect on the production of ammonia, H2S, or skatole in animals fed Bs. In both groups, fructan was the principle additive that reduced H2S production (r = 0.9981). Fructan and Bs significantly interacted in H2S production (p = 0.014). Casein was the principle additive affecting the concentration of ammonia, only in the control group. Casein and Bs significantly interacted in ammonia production (p = 0.039). The predominant bacteria were Bacillus spp. CWBI B1434 (26%) in the control group, and Streptococcus alactolyticus AF201899 (36%) in the experimental group. In summary, daily dietary Bs reduced ammonia production during fecal fermentation. Lessening L-tryptophan and increasing fructan in the fermentation broth reduced skatole and H2S.

Genetic Variation of H-FABP Gene and Association with Intramuscular Fat Content in Laiwu Black and Four Western Pig Breeds

Zeng, Y.Q.,Wang, G.L.,Wang, C.F.,Wei, S.D.,Wu, Y.,Wang, L.Y.,Wang, H.,Yang, H.L. Asian Australasian Association of Animal Productio 2005 Animal Bioscience Vol.18 No.1

This study was performed to detect genetic variation of the heart fatty acid-binding protein (H-FABP) gene by PCRRFLPs approach and its association with intramuscular fat (IMF) content. Data from 223 individuals, including one Chinese native pig breed and four western pig breeds, were analyzed. The results showed that for the H-FABP gene, there was one polymorphic HinfI site in the 5'-upstream region, whereas there were one HaeIII and one HinfI (marked as $HinfI^*$) polymorphic site in the second intron, respectively. The three PCR-RFLPs were present in all breeds tested. The allele frequencies, however, revealed significant differences between them (p<0.05). Furthermore, the allele frequency distribution of HinfI in the Laiwu Black and that of $HinfI^*$ in the Hampshire breed were at disequilibrium, which might be the result of selective breeding. Results also indicated that for HinfI, HaeIII and $HinfI^*$ HFABP RFLP, significant (p<0.05) contrasts of 0.78%, -0.69% and 0.72% were detected in the least square means of IMF content between the homozygous genotype HH and hh, DD and dd, BB and bb classes, respectively. It implied that the HHddBB genotype had the highest IMF content in this experimental population and these H-FABP RFLPs could serve, to some extent, as genetic markers for use in improvement of IMF content.

The evolutionary dynamics of highly pathogenic avian influenza H5N1 in south-central Vietnam reveals multiple clades evolving from Chinese and Cambodian viruses

Nguyen, T.H.,Than, V.T.,Thanh, H.D.,Nguyen, V.Q.,Nguyen, K.H.,Nguyen, D.T.,Park, J.H.,Chung, I.S.,Jeong, D.G.,Chang, K.T.,Oh, T.K.,Kim, W. Pergamon Press 2015 Comparative immunology, microbiology and infectiou Vol.42 No.-

In Vietnam, highly pathogenic avian influenza (HPAI), such as that caused by H5N1 viruses, is the most highly contagious infectious disease that has been affecting domestic poultry in recent years. Vietnam might be an evolutionary hotspot and a potential source of globally pandemic strains. However, few studies have reported viruses circulating in the south-central region of Vietnam. In the present study, 47 H5N1-positive samples were collected from both vaccinated and unvaccinated poultry farms in the South Central Coast region of Vietnam during 2013-2014, and their genetic diversity was analyzed. A common sequence motif for HPAI virus was identified at HA-cleavage sites in all samples: either RERRRKR/G (clades 2.3.2.1c and 2.3.2.1a) or REGRRKKR/G (clade 1.1.2). Phylogenetic analysis of HA genes identified three clades of HPAI H5N1: 1.1.2 (n=1), 2.3.2.1a (n=1), and 2.3.2.1c (n=45). The phylogenetic analysis indicated that these Vietnamese clades may have evolved from Chinese and Cambodian virus clades isolated in 2012-2013 but are less closely related to the clades detected from the Tyva Republic, Bulgaria, Mongolia, Japan, and Korea in 2009-2011. Detection of the coexistence of virus clades 2.3.2.1 and the very virulent 1.1.2 in the south-central regions suggests their local importance and highlights concerns regarding their spread, both northwards and southwards, as well as the potential for reassortment. The obtained data highlight the importance of regular identification of viral evolution and the development and use of region-specific vaccines.

Emergence of Amantadine-Resistant H3N2 Avian Influenza A Virus in South Korea

Lee, J.,Song, Y. J.,Park, J. H.,Lee, J.-H.,Baek, Y. H.,Song, M.-S.,Oh, T.-K.,Han, H.-S.,Pascua, P. N. Q.,Choi, Y.-K. American Society for Microbiology 2008 Journal of clinical microbiology Vol.46 No.11

<P>We found a relatively high frequency of unique amantadine-resistant H3N2 and H9N2 avian influenza viruses (Val27Ile on M2 protein) isolated from live poultry markets in South Korea and confirmed that a Val27Ile single substitution in the M2 protein is enough to acquire the amantadine resistance phenotype by using reverse-genetically created human-avian reassortant viruses.</P>

Cloning, characterization and tissue expression of disk abalone (Haliotis discus discus) catalase

Ekanayake, P.M.,De Zoysa, M.,Kang, H.S.,Wan, Q.,Jee, Y.,Lee, Y.H.,Kim, S.J.,Lee, J. Academic Press 2008 FISH AND SHELLFISH IMMUNOLOGY Vol.24 No.3

Catalase is an antioxidant enzyme that plays a significant role in protection against oxidative stress by detoxification of hydrogen peroxide (H<SUB>2</SUB>O<SUB>2</SUB>). A gene coding for a putative catalase was isolated from the disk abalone (Haliotis discus discus) cDNA library and denoted as Ab-catalase. The full-length (2864bp) Ab-catalase cDNA contained 1,503bp open reading frame (ORF), encoding 501 amino acid residues with 56kDa predicted molecular weight. The deduced amino acid sequence of Ab-catalase has characteristic features of catalase family such as catalytic site motif (<SUP>61</SUP>FNRERIPERVVHAKGAG<SUP>77</SUP>), heme-ligand signature motif (<SUP>351</SUP>RLYSYSDT<SUP>358</SUP>), NADPH and heme binding residues. Phylogenetic and pairwise identity results indicated that Ab-catalase is more similar to scallop (Chlamys farreri) catalase with 80% amino acid identity except for other reported disk abalone catalase sequences. Constitutive Ab-catalase expression was detected in gill, mantle, gonad, hemocytes, abductor muscle and digestive tract in tissue specific manner. Ab-catalase mRNA was up-regulated in gill and digestive tract tissues for the first 3h post injection of H<SUB>2</SUB>O<SUB>2</SUB>, showing the inducible ability of abalone catalase against oxidative stress generated by H<SUB>2</SUB>O<SUB>2</SUB>. The purified recombinant catalase showed 30,000U/mg enzymatic activity against H<SUB>2</SUB>O<SUB>2</SUB> and biochemical properties of higher thermal stability and broad spectrum of pH. Our results suggest that abalone catalase may play an important role in regulating oxidative stress by scavenging H<SUB>2</SUB>O<SUB>2</SUB>.

Hydrogen peroxide-responsive copolyoxalate nanoparticles for detection and therapy of ischemia-reperfusion injury

Lee, D.,Bae, S.,Ke, Q.,Lee, J.,Song, B.,Karumanchi, S.A.,Khang, G.,Choi, H.S.,Kang, P.M. Elsevier Science Publishers 2013 Journal of controlled release Vol.172 No.3

The main culprit in the pathogenesis of ischemia/reperfusion (I/R) injury is the generation of high level of hydrogen peroxide (H<SUB>2</SUB>O<SUB>2</SUB>). In this study, we report a novel diagnostic and therapeutic strategy for I/R injury based on H<SUB>2</SUB>O<SUB>2</SUB>-activatable copolyoxalate nanoparticles using a murine model of hind limb I/R injury. The nanoparticles are composed of hydroxybenzyl alcohol (HBA)-incorporating copolyoxalate (HPOX) that, in the presence of H<SUB>2</SUB>O<SUB>2</SUB>, degrades completely into three known and safe compounds, cyclohexanedimethanol, HBA and CO<SUB>2</SUB>. HPOX effectively scavenges H<SUB>2</SUB>O<SUB>2</SUB> in a dose-dependent manner and hydrolyzes to release HBA which exerts intrinsic antioxidant and anti-inflammatory activities both in vitro and in vivo models of hind limb I/R. HPOX nanoparticles loaded with fluorophore effectively and robustly image H<SUB>2</SUB>O<SUB>2</SUB> generated in hind limb I/R injury, demonstrating their potential for bioimaging of H<SUB>2</SUB>O<SUB>2</SUB>-associated diseases. Furthermore, HPOX nanoparticles loaded with anti-apoptotic drug effectively release the drug payload after I/R injury, exhibiting their effectiveness for a targeted drug delivery system for I/R injury. We anticipate that multifunctional HPOX nanoparticles have great potential as H<SUB>2</SUB>O<SUB>2</SUB> imaging agents, therapeutics and drug delivery systems for H<SUB>2</SUB>O<SUB>2</SUB>-associated diseases.

China Spallation Neutron Source: Accelerator Design Iterations and R&D Status

J. Wei,C.-D. Deng,C.-H. Wang,C.-T. Shi,H. Sun,H.-F. Ouyang,H.-M. Qu,H.-Y. Dong,J. Li,J. Zhang,J.-S. Cao,J.-Y. Tang,L. Dong,L.-L. Wang,Q. Qin,Q.-B. Wang,S. Wang,S.-N. Fu,S.-X Fang,T. -G. Xu,W. Kang,Y.- 한국물리학회 2007 THE JOURNAL OF THE KOREAN PHYSICAL SOCIETY Vol.50 No.I

The China Spallation Neutron Source (CSNS) is a high-power, accelerator-based project currently under preparation. The accelerator complex consists of an H$^-$ ion source, an H$^-$ linac, a rapid-cycling proton synchrotron, and the transport lines. During the past year, the design of most accelerator systems went through major iterations, and initial research and developments was started on the prototyping of several key components.