A C1 inhibitor ortholog from rock bream (Oplegnathus fasciatus): Molecular perspectives of a central regulator in terms of its genomic arrangement, transcriptional profiles and anti-protease activities of recombinant peptide

Umasuthan, N.,Bathige, S.D.N.K.,Revathy, K.S.,Wickramaarachchi, W.D.N.,Wan, Q.,Whang, I.,Kim, E.,Park, M.A.,Park, H.C.,Lee, J. Pergamon Press ; Elsevier Science 2014 DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY Vol.42 No.2

C1 inhibitor (C1Inh), a member of serpin superfamily, is a crucial regulator of the activation of various plasmatic cascades associated with immunity and inflammation. This study describes the identification and characterization of a C1Inh gene from rock bream Oplegnathus fasciatus (OfC1Inh) at structural, expressional and functional levels. The cDNA-(2245bp) and corresponding gDNA-sequences (5.2kbp) of OfC1Inh were isolated from rock bream transcriptome- and BAC-libraries, respectively. Predicted amino acid sequence of OfC1Inh revealed a two-domain architecture composed of an N-terminal region with two Ig-like domains and a C-terminal region with a serpin domain. Tertiary model of OfC1Inh disclosed its active site topology. In the multi-exonic genomic arrangement of OfC1Inh, it consisted of eleven exons disjoined by ten introns as observed in few other fish homologs. Our comparative analysis indicated that the teleostean C1Inhs were distinct from their non-teleostean vertebrate counterparts in terms of their (1) extended N-terminal domains, (2) evolutionary divergence and (3) exon-intron distribution. The OfC1Inh had a TATA-deficient promoter with a putative initiator element, and two tandemly arranged downstream promoter elements. Several components associated with the immune and inflammatory transcriptional activation were also predicted to exist in 5' flanking region of OfC1Inh. The exclusive mRNA levels in liver and moderate levels in extra-hepatic tissues intimated the diversified importance of OfC1Inh in rock bream physiology. We also provide an evidence for the involvement of OfC1Inh in immune balance, based on its modulated transcription upon different PAMP (lipopolysaccharide and poly I:C)- or pathogen (Streptococcus iniae and rock bream irido virus)-challenges. A recombinantly expressed fusion protein [(r)OfC1Inh] was employed in demonstrating the anti-protease function of OfC1Inh. The (r)OfC1Inh exhibited detectable inhibitory activity against C1 esterase and thrombin, where the anti-C1 esterase role was shown to be potentiated by heparin. Taken together, the results of this study provide the first line of evidence for the possible involvement of a teleostean C1Inh in fish immunity, based on its expressional response(s) and inhibitory properties against two enzymes involved in biological cascades.

An albumin nanocomplex-based endosomal pH-activatable on/off probe system

Lee, C.,Lee, S.,Thao, L.Q.,Hwang, H.S.,Kim, J.O.,Lee, E.S.,Oh, K.T.,Shin, B.S.,Choi, H.G.,Youn, Y.S. Elsevier 2016 Colloids and Surfaces B Vol.144 No.-

<P>Albumin has gained considerable interest as a material for fabricating nanoparticulate systems due to its biomedical advantages, such as biocompatibility and chemical functionality. Here, we report a new pH-sensitive albumin nanocomplex prototype with a zinc-imidazole coordination bond. Albumin was conjugated with 1-(3-aminopropyl)imidazole and mPEG(10kDa)-NHS, and the resulting albumin conjugate (PBI) was then modified with either Cy5.5 or BHQ-3. The newly formed albumin nanocomplex (C/BQ-PBI Zn NCs: similar to 116 nm) system was facilely self-assembled around pH 7.4 in the presence of Zn2+, but it quickly disassembled in an acidic environment (similar to pH 5.0). Based on this pH-sensitivity, C/BQ-PBI Zn NCs emitted strong near-infrared fluorescence and released Zn2+, turning 'off' at pH similar to 7.4 (e.g., plasma) and 'on' at pH similar to 5.0 (e.g., endo/lysosomes in tumor cells) on account of fluorescence resonance energy transfer. C/BQ-PBI Zn NCs displayed significant cytotoxicity due to an increase in cellular. Zn2+ in response to endosomal pH (similar to 5.0) in breast cancer MCF-7 cells and lung adenocarcinoma A549 cells. Particularly, confocal laser scanning microscopic images showed a strong fluorescence signal caused by the disassembly of C/BQ-PBI Zn NCs in the endosomal region of MCF-7 cells. Based on these results, we believe that this albumin nanocomplex is an attractive biocompatible tumor targeting probe carrier for the theranostic purpose. (C) 2016 Elsevier B.V. All rights reserved.</P>

Genomic characterization and expression analysis of complement component 9 in rock bream (Oplegnathus fasciatus)

Wickramaarachchi, W.D.N.,Wan, Q.,Lee, Y.,Lim, B.S.,De Zoysa, M.,Oh, M.J.,Jung, S.J.,Kim, H.C.,Whang, I.,Lee, J. Academic Press 2012 Fish & Shellfish Immunology Vol.33 No.4

The complement component 9 (C9) is a single-chain glycoprotein that mediates formation of the membrane attack complex (MAC) on the surface of target cells. Full-length C9 sequence was identified from a cDNA library of rock bream (Oplegnathus fasciatus), and its genomic sequence was obtained by screening and sequencing of a bacterial artificial chromosome (BAC) genomic DNA library of rock bream. The rock bream complement component 9 (Rb-C9) gene contains 11 exons and 10 introns and is composed of a 1782 bp complete open reading frame (ORF) that encodes a polypeptide of 593 amino acids. Sequence analysis revealed that the Rb-C9 protein contains two thrombospondin type-1domains, a low-density lipoprotein receptor domain class A, a membrane attack complex & perforin (MACPF) domain, and an epidermal growth factor (EGF)-like domain. Important putative transcription factor binding sites, including those for NF-κB, SP-1, C/EBP, AP-1 and OCT-1, were found in the 5' flanking region. Phylogenetic analysis revealed a close proximity of Rb-C9 with the orthologues in puffer fish, and Japanese flounder. Quantitative real-time RT-PCR analysis confirmed that Rb-C9 was constitutively expressed in all the examined tissues isolated from healthy rock bream, with highest expression occurring in liver. Pathogen challenge, including Edwardsiella tarda, Streptococcus iniae, lipopolysaccharide endotoxin and rock bream iridovirus led to up-regulation of Rb-C9 in liver but no change in peripheral blood cells. The observed response to bacterial and viral challenges and high degree of evolutionary relationship to respective orthologues, confirmed that Rb-C9 is an important immune gene, likely involved in the complement system lytic pathway of rock bream.

Bioconversion of ginsenosides Rb₁, Rb₂, Rc and Rd by novel β-glucosidase hydrolyzing outer 3-O glycoside from Sphingomonas sp. 2F2: Cloning, expression, and enzyme characterization

Wang, L.,Liu, Q.M.,Sung, B.H.,An, D.S.,Lee, H.G.,Kim, S.G.,Kim, S.C.,Lee, S.T.,Im, W.T. Elsevier Science Publishers 2011 Journal of biotechnology Vol.156 No.2

A new β-glucosidase gene (bglSp) was cloned from the ginsenoside converting Sphingomonas sp. strain 2F2 isolated from the ginseng cultivating filed. The bglSp consisted of 1344bp (447 amino acid residues) with a predicted molecular mass of 49,399Da. A BLAST search using the bglSp sequence revealed significant homology to that of glycoside hydrolase superfamily 1. This enzyme was overexpressed in Escherichia coli BL21 (DE3) using a pET21-MBP (TEV) vector system. Overexpressed recombinant enzymes which could convert the ginsenosides Rb<SUB>1</SUB>, Rb<SUB>2</SUB>, Rc and Rd to the more pharmacological active rare ginsenosides gypenoside XVII, ginsenoside C-O, ginsenoside C-Mc<SUB>1</SUB> and ginsenoside F<SUB>2</SUB>, respectively, were purified by two steps with Amylose-affinity and DEAE-Cellulose chromatography and characterized. The kinetic parameters for β-glucosidase showed the apparent K<SUB>m</SUB> and V<SUB>max</SUB> values of 2.9+/-0.3mM and 515.4+/-38.3μmolmin<SUP>-1</SUP>mg of protein<SUP>-1</SUP> against p-nitrophenyl-β-d-glucopyranoside. The enzyme could hydrolyze the outer C3 glucose moieties of ginsenosides Rb<SUB>1</SUB>, Rb<SUB>2</SUB>, Rc and Rd into the rare ginsenosides Gyp XVII, C-O, C-Mc<SUB>1</SUB> and F<SUB>2</SUB> quickly at optimal conditions of pH 5.0 and 37<SUP>o</SUP>C. A little ginsenoside F<SUB>2</SUB> production from ginsenosides Gyp XVII, C-O, and C-Mc<SUB>1</SUB> was observed for the lengthy enzyme reaction caused by the side ability of the enzyme.

The Efficacy of Enhanced Growth by Ectopic Expression of Ghrelin and Its Variants Using Injectable Myogenic Vectors

Xie, Q.F.,Wu, C.X.,Meng, Q.Y.,Li, N. Asian Australasian Association of Animal Productio 2004 Animal Bioscience Vol.17 No.1

Ghrelin is an acylated peptide recently identified as the endogenous ligand for the growth hormone (GH) secretagogues receptor 1a (GHS-R1a) and is involved in a novel system for regulating GH release. To understand the long-term effects of ghrelin, here we constructed six myogenic expression vectors containing the cDNA of swine mature ghrelin (pGEM-wt-sGhln, pGEM-wt-hGhln), ghrelin mutant of $Ser^3$ with $Trp^3$ (pGEM-mt-sGhln, pGEM-mt-hGhln) and truncated ghrelin derivative (pGEM-tmtsGhln, pGEM-tmt-hGhln) encompassing the first 7 residues of ghrelin (including $Ser^3$ substituted with $Trp^3$) and adding a basic amino acid, Lys (K) in the C-terminus. The constructs, pGEM-wt-sGhln, pGEM-mt-sGhln and pGEM-tmt-sGhln were linked with the ghrelin leader sequence, while the pGEM-wt-hGhln, pGEM-mt-hGhln and pGEM-tmt-hGhln were linked with a leader sequence from the human growth hormone releasing hormone (hGHRH). Intramuscular injection of 200 ${\mu}g$ pGEM-wt-sGhln or pGEM-tmt-sGhln augmented growth over 3 weeks in normal rats and peaked at day 21 or 14 post-injection respectively, whose body weight gains were on average approximately 6% or 19% heavier over controls. However, other injectable vectors had no such enhanced growth effects. Our results suggested that the efficacy of the ghrelin leader sequence was more effective than that of hGHRH in our system. Moreover, the results indicated that skeletal muscle might have the ability to posttranslationally modify the in vivo expressed ghrelin. And the most strikingly, the short ghrelin analog seems to mimic the biological effects more efficiently when compared with the full-length ghrelin.

Fabricating Ultrathin Plate-Like WC Grains in WC–8Co Hardmetals by Increasing Discharge Intensity During Plasma-Assisted Ball Milling

M. Q. Zeng,J. L. Tu,M. Zhu,W. Wang,J. W. Liu,Z. C. Lu 대한금속·재료학회 2020 METALS AND MATERIALS International Vol.26 No.9

The effects of plasma discharge intensity on the microstructure evolution of ball-milled tungsten (W)–carbon (C)–cobalt (Co)mixtures and the formation mechanism of ultrathin plate-like tungsten carbide (WC) grains prepared by ball milling with andwithout plasma discharge were investigated. It was found that increasing the plasma discharge intensity during ball millingobviously promoted the formation of a thin flake-like W phase because of the electroplasticity effect and simultaneouslylowered the carburization temperature between W and C. A combination of high hardness and transverse rupture strength of92.9 HRA and 3659 MPa, respectively, was obtained for the WC–8Co alloy fabricated by plasma milling at a gas pressure of5 × 103 Pa with a dielectric barrier discharge layer thickness of 3 mm. These properties were mainly attributed to the markedlylowered activation energy of the WC phase and generation of highly oriented ultrathin plate-like WC grains by plasma milling. The combination of the flake-like structure of the plasma-milled W aggregate and high specific interfacial area and shortdiffusion distance of W/C were readily inherited by the ultrathin plate-like WC grains in the sintered WC–Co hardmetals.

Evaluation of VIIRS, GOCI, and MODIS Collection 6 AOD retrievals against ground sunphotometer observations over East Asia

Xiao, Q.,Zhang, H.,Choi, M.,Li, S.,Kondragunta, S.,Kim, J.,Holben, B.,Levy, R. C.,Liu, Y. Copernicus GmbH 2016 Atmospheric Chemistry and Physics Vol.16 No.3

<P>Abstract. Persistent high aerosol loadings together with extremely high population densities have raised serious air quality and public health concerns in many urban centers in East Asia. However, ground-based air quality monitoring is relatively limited in this area. Recently, satellite-retrieved Aerosol Optical Depth (AOD) at high resolution has become a powerful tool to characterize aerosol patterns in space and time. Using ground AOD observations from the Aerosol Robotic Network (AERONET) and the Distributed Regional Aerosol Gridded Observation Networks (DRAGON)-Asia Campaign, as well as from handheld sunphotometers, we evaluated emerging aerosol products from the Visible Infrared Imaging Radiometer Suite (VIIRS) aboard the Suomi National Polar-orbiting Partnership (S-NPP), the Geostationary Ocean Color Imager (GOCI) aboard the Communication, Ocean, and Meteorology Satellite (COMS), and Terra and Aqua Moderate Resolution Imaging Spectroradiometer (MODIS) (Collection 6) in East Asia in 2012 and 2013. In the case study in Beijing, when compared with AOD observations from handheld sunphotometers, 51 % of VIIRS Environmental Data Record (EDR) AOD, 37 % of GOCI AOD, 33 % of VIIRS Intermediate Product (IP) AOD, 26 % of Terra MODIS C6 3 km AOD, and 16 % of Aqua MODIS C6 3 km AOD fell within the reference expected error (EE) envelope (±0.05 ± 0.15 AOD). Comparing against AERONET AOD over the Japan-South Korea region, 64 % of EDR, 37 % of IP, 61 % of GOCI, 39 % of Terra MODIS, and 56 % of Aqua MODIS C6 3 km AOD fell within the EE. In general, satellite aerosol products performed better in tracking the day-to-day variability than tracking the spatial variability at high resolutions. The VIIRS EDR and GOCI products provided the most accurate AOD retrievals, while VIIRS IP and MODIS C6 3 km products had positive biases. </P>

MK-801, but not naloxone, attenuates high-dose dextromethorphan-induced convulsive behavior: Possible involvement of the GluN2B receptor

Tran, H.Q.,Chung, Y.H.,Shin, E.J.,Tran, T.V.,Jeong, J.H.,Jang, C.G.,Nah, S.Y.,Yamada, K.,Nabeshima, T.,Kim, H.C. Academic Press 2017 Toxicology and applied pharmacology Vol.334 No.-

Dextromethorphan (DM) is a dextrorotatory isomer of levorphanol, a typical morphine-like opioid. When administered at supra-antitussive doses, DM produces psychotoxic and neurotoxic effects in humans. Although DM abuse has been well-documented, few studies have examined the effects of high-dose DM. The present study aimed to explore the effects of a single high dose of DM on mortality and seizure occurrence. After intraperitoneal administration with a high dose of DM (80mg/kg), Sprague-Dawley rats showed increased seizure occurrence and intensity. Hippocampal expression levels of N-methyl-d-aspartate (NMDA) receptor subunits (GluN1<GluN2A<GluN2B), c-Fos and pro-apoptotic factors (Bax and cleaved caspase-3) were upregulated by DM treatment; while levels of anti-apoptotic factors (Bcl-2 and Bcl-xL) were downregulated. Consistently, DM also induced ultrastructural degeneration in the hippocampus. A non-competitive NMDA receptor antagonist, MK-801, attenuated these effects of high-dose DM, whereas an opioid antagonist, naloxone, did not affect DM-induced neurotoxicity. Moreover, pretreatment with a highly specific GluN2B subunit inhibitor, traxoprodil, was selectively effective in preventing DM-induced c-Fos expression and apoptotic changes. These results suggest that high-dose DM produces convulsive behaviors by activating GluN2B/NMDA signaling that leads to pro-apoptotic changes.

Association Analyses with Carcass Traits in the Porcine KIAA1717 and HUMMLC2B Genes

Xu, D.Q.,Xiong, Y.Z.,Liu, M.,Lan, J.,Ling, X.F.,Deng, C.Y.,Jiang, S.W. Asian Australasian Association of Animal Productio 2005 Animal Bioscience Vol.18 No.11

By screening a subtracted cDNA library constructed with mRNA obtained from the longissimus dorsi muscles of F1 hybrids Landrace${\times}$Yorkshire and their Yorkshire female parents, we isolated two partial sequences coding for the H3-K4-specific methyltransferase (KIAA1717) and skeletal muscle myosin regulatory light chain (HUMMLC2B) genes. In the present work we investigated two SNPs, one (C1354T) at the 3' untranslated region (UTR) of KIAA1717 and one (A345G) at the SINE (PRE-1) element of HUMMLC2B, in a resource population derived from crossing Chinese Meishan and Large White pig. The selected pigs were genotyped by means of a PCR-RFLP protocol. Significant associations were observed for the KIAA1717 C1354T polymorphic site with thorax-waist backfat thickness (p<0.05), buttock backfat thickness (p<0.05), average backfat thickness (p<0.05), loin eye height (p<0.05), loin eye area (p<0.05), carcass length to 1$^{st}$ spondyle (p<0.01) and carcass length to 1st rib (p<0.01). HUMMLC2B A345G were significantly associated with loin eye width (p<0.05), loin eye area (p<0.05). Further studies are needed to confirm these preliminary results.

Comparative Palm Oil Trade Performance in Indonesia, Malaysia, and the Philippines

Maria Noriza Q,Herrera,Renen Szilardo C,de Guzman,Dinah Pura T,Depositario,Loida E,Mojica,Jeanette Angelin B,Madamba The International Academy of Global Business and T 2019 The International Academy of Global Business and T Vol.15 No.2

Purpose – As one of the most traded commodities in the world with tremendous economic significance, palm oil is an inexpensive source of oil. This paper examined and compared the palm oil trade performance of the Philippines relative to its top palm oil producing neighbors, Indonesia and Malaysia, by analyzing importexport volume trends of the said food commodity, consumption and production trends, self-sufficiency, import dependence, and shifting patterns of comparative advantage. Design/Methodology/Approach – Secondary data covering the period from 1964 through 2018 were collected from various sources, and descriptive analysis coupled with trend analysis were employed with respect to import and export, production, and consumption performance. Other trade indicators were computed, to wit: balance of trade, self-sufficiency ratio, import dependency ratio, and normalized trade balance. Implications were drawn after the comparative analysis among the three countries was done. Findings – An increase in the volume of the palm oil supply in the Philippines is evident, and this is attributed to increasing importation. Relative to Malaysia and Indonesia, the Philippines lags behind in international trade. While Philippine palm oil industry growth has been stagnant in terms of international trade by virtue of its net importer status, Malaysia and Indonesia have overtaken the country significantly, having attained selfsufficiency in the commodity with trade surpluses, and the gap is expected to widen in the coming years. It is interesting to note that the Philippines, Malaysia, and Indonesia had identical production volumes and growth rates until the last decade of the 21st century. A steadily growing industry, palm oil is foreseen to continue being a vital industry, not only to top producing countries such as Malaysia and Indonesia but also in the Philippines, which is trying to improve its palm oil trade performance by expanding production. Research Implications – The study findings imply expansion and income opportunities not only for local oil palm producers, processors, and traders in the Philippines but also for foreign market entrants.