Restricted growth of U‐type infectious haematopoietic necrosis virus (IHNV) in rainbow trout cells may be linked to casein kinase II activity

Park, J W,Moon, C H,Harmache, A,Wargo, A R,Purcell, M K,Bremont, M,Kurath, G Blackwell Publishing Ltd 2011 Journal of fish diseases Vol.34 No.2

<P><B>Abstract</B></P><P>Previously, we demonstrated that a representative M genogroup type strain of infectious haematopoietic necrosis virus (IHNV) from rainbow trout grows well in rainbow trout‐derived RTG‐2 cells, but a U genogroup type strain from sockeye salmon has restricted growth, associated with reduced genome replication and mRNA transcription. Here, we analysed further the mechanisms for this growth restriction of U‐type IHNV in RTG‐2 cells, using strategies that assessed differences in viral genes, host immune regulation and phosphorylation. To determine whether the viral glycoprotein (G) or non‐virion (NV) protein was responsible for the growth restriction, four recombinant IHNV viruses were generated in which the G gene of an infectious IHNV clone was replaced by the G gene of U‐ or M‐type IHNV and the NV gene was replaced by NV of U‐ or M‐type IHNV. There was no significant difference in the growth of these recombinants in RTG‐2 cells, indicating that G and NV proteins are not major factors responsible for the differential growth of the U‐ and M‐type strains. Poly I:C pretreatment of RTG‐2 cells suppressed the growth of both U‐ and M‐type IHNV, although the M virus continued to replicate at a reduced level. Both viruses induced type 1 interferon (IFN1) and the IFN1 stimulated gene Mx1, but the expression levels in M‐infected cells were significantly higher than in U‐infected cells and an inhibitor of the IFN1‐inducible protein kinase PKR, 2‐aminopurine (2‐AP), did not affect the growth of U‐ or M‐type IHNV in RTG‐2 cells. These data did not indicate a role for the IFN1 system in the restricted growth of U‐type IHNV in RTG‐2 cells. Prediction of kinase‐specific phosphorylation sites in the viral phosphoprotein (P) using the NetPhosK program revealed differences between U‐ and M‐type P genes at five phosphorylation sites. Pretreatment of RTG‐2 cells with a PKC inhibitor or a p38MAPK inhibitor did not affect the growth of the U‐ and M‐type viruses. However, 100 μ<SMALL>m</SMALL> of the casein kinase II (CKII) inhibitor, 5,6‐dichloro‐1‐β‐<SMALL>d</SMALL>‐ribofuranosylbenzimidazole (DRB), reduced the titre of the U type 8.3‐fold at 24 h post‐infection. In contrast, 100 μ<SMALL>m</SMALL> of the CKII inhibitor reduced the titre of the M type only 1.3‐fold at 48 h post‐infection. Our data suggest that the different growth of U‐ and M‐type IHNV in RTG‐2 cells may be linked to a differential requirement for cellular protein kinases such as CKII for their growth.</P>

Role of flotillins in the endocytosis of GPCR in salivary gland epithelial cells

Park, M.Y.,Kim, N.,Wu, L.L.,Yu, G.Y.,Park, K. Academic Press 2016 Biochemical and biophysical research communication Vol.476 No.4

Endocytosis has numerous functions in cellular homeostasis. Defects in the endocytic pathway of receptors may lead to dysfunction of salivary gland secretion. Therefore, elucidating the complex mechanisms of endocytosis may facilitate solutions for disease treatment and prevention. The muscarinic type 3 receptor (M3R), a G-protein-coupled receptor (GPCR) located in the plasma membrane, is involved in numerous physiological activities such as smooth muscle contraction and saliva secretion. M3R enters cells through clathrin-mediated endocytosis (CME), while flotillins (flot-1 and -2), highly conserved proteins residing in lipid-raft microdomains, make use of clathrin-independent endocytosis (CIE) for their internalization. Since these two proteins use two discrete pathways for endocytic entry, the association of flotillins with CME is poorly understood. We examined whether flotillins play a role in CME of M3R using immunoblotting, immunocytochemistry, confocal immunofluorescence microscopy, co-immunoprecipitation, and RNA interference techniques in secretory epithelial cells. Upon stimulation with a cholinergic agonist, M3R, flot-1, and flot-2 each internalized from the plasma membrane into intracellular sites. The addition of chlorpromazine and cytochalasin D, well-known inhibitors of CME, inhibited internalization of M3R via CME. Filipin III and methyl-β-cyclodextrin (mβCD) acting as lipid raft inhibitors disrupted internalization of flot-½ via CIE. Interestingly, filipin III and mβCD slightly reduced expression level of M3R whereas chlorpromazine and cytochalasin D did not affect internalization of the flotillin isoforms. M3R and flot-½ colocalized and interacted with each other as they entered the cytosol during limited periods of incubation. Moreover, knockdown of flot-1 or -2 by flotillin-specific siRNA prevented internalization and reduced the endocytic efficiency of M3R. Our results suggest that flot-1 and -2 are partially involved in CME of M3R by facilitating its internalization.

미세각막절개도 헤드의 선택을 통한 맞춤식각막절개방식의 라식

강민호,송유미,강성민,박영숙,최철영,이윤정,이병로.Min Ho Kang. M.D.. Yoo Mi Song. M.D.. Sung Min Kang. M.D.. Young Sook Park. M.D.. Chul Young Choi. M.D.. Yoon Jung Lee. M.D.. Byung Ro Lee. M.D. 대한안과학회 2006 대한안과학회지 Vol.47 No.2

Purpose: To compare the intraoperative and postoperative complication rates and visual outcomes with M2 130 head (thick flap group) and M2 110 head (thin flap group). Methods: One-hundred-ninety-five eyes of 104 patients who underwent LASIK with the Moria M2 microkeratome and Allegretto-wave laser were reviewed retrospectively. Selection of M2 heads was based on preoperative pachymetry and estimated ablation depth. Intraoperative and postoperative flap-related complications, mean postoperative uncorrected visual acuity (UCVA), and mean spherical equivalent (MSE) were evaluated and compared. Results: Mean follow-up was 8.1 months. The number of eyes, preoperative MSE and mean corneal thickness of M2 130 were 115, -4.04±1.63diopter (D) and 549.40±39.16??m, and 85, -6.61±3.43D and 525.16±24.53 ?m, respectively, in the 110 head group. Mean UCVA and MSE at postoperative 1 week, 3 months and 6 months were 1.00±0.18, -0.26±0.49D; 1.07±0.68, -0.40±0.51D; and 1.01±0.22, -0.51±0.50D, respectively, in the 130 head group, and 0.90±0.23, -0.46±1.02D; 0.91±0.23, -0.67±0.79D; 0.85±0.46, -0.75±0.88D, respectively, in the 110 head group. There were no significant differences in intraoperative and postoperative flap-related complication rates between the two groups (p=0.316). There were no statistically significant differences in postoperative mean UCVA or MSE between the two groups (p>0.05), except for MSE at the third and sixth postoperative months (p=0.005, 0.013). Conclusions: Proper selection of M2 heads by preoperative pachymetry allowed for an adequate residual stroma bed with good visual outcome. Utilizing one single microkeratome and switching between two heads was advantageous and cost-effective.

High-resolution metabolomics to identify urine biomarkers in corticosteroid-resistant asthmatic children

Park, Youngja H.,Fitzpatrick, Anne M.,Medriano, Carl Angelo,Jones, Dean P. Elsevier 2017 The Journal of allergy and clinical immunology Vol.139 No.5

<P><B>Background</B></P> <P>Corticosteroid (CS) treatment has been established as the first anti-inflammatory treatment for adults and children with asthma. However, a subset of patients fails to respond to combined systemic and inhaled CS treatment.</P> <P><B>Objective</B></P> <P>This study was aimed at further understanding CS resistance among children with severe asthma.</P> <P><B>Methods</B></P> <P>High-resolution metabolomics was performed on urine samples from CS-respondent (n = 15) and CS-nonrespondent (n = 15) children to determine possible urine biomarkers related to CS resistance. The metabolic phenotypes of CS responders and CS nonresponders were analyzed using bioinformatics including Manhattan plot with false- discovery rate, hierarchical cluster analysis, Kyoto Encyclopedia Genes and Genomes, and Mummichog pathway analysis.</P> <P><B>Results</B></P> <P>The 2-way hierarchical cluster analysis study determined 30 metabolites showing significantly different levels between CS responders and CS nonresponders. The important metabolites annotated were 3,6-dihydronicotinic acid (126.05 <I>m</I>/<I>z</I>, RT: 106, [M+H]<SUP>+</SUP>), 3-methoxy-4-hydroxyphenyl(ethylene)glycol (185.05 <I>m</I>/<I>z</I>, RT: 155, [M+H]<SUP>+</SUP>), 3,4-dihydroxy-phenylalanine (198.07 <I>m</I>/<I>z</I>, RT: 446, [M+H]<SUP>+</SUP>), γ-glutamylcysteine (236.06 <I>m</I>/<I>z</I>, RT: 528, [M+S(34)+H]<SUP>+</SUP>), Cys-Gly, (253.06 <I>m</I>/<I>z</I>, RT: 528, [M-NH<SUB>3</SUB>+H]<SUP>+</SUP>), and reduced Flavin mononucleotide (517.0794 <I>m/z</I>, RT: 533, [M+NaCl]<SUP>+</SUP>). Tyrosine metabolism, degradation of aromatic compounds, and glutathione metabolism are suggested to be significant pathways relating to CS resistance.</P> <P><B>Conclusions</B></P> <P>High-resolution metabolomics is a promising approach in asthma research. Five candidate markers were identified to be related to CS-resistant children with severe asthma. These compounds, upon validation, may contribute further in the understanding of CS resistance among children with severe asthma through the use of urine.</P>

Supplementation of oil-based inactivated H9N2 vaccine with M2e antigen enhances resistance against heterologous H9N2 avian influenza virus infection

Park, J.K.,Lee, D.H.,Cho, C.H.,Yuk, S.S.,To, E.O.,Kwon, J.H.,Noh, J.Y.,Kim, B.Y.,Choi, S.W.,Shim, B.S.,Song, M.K.,Lee, J.B.,Park, S.Y.,Choi, I.S.,Song, C.S. Elsevier Scientific Pub. Co 2014 Veterinary microbiology Vol.169 No.3

Avian influenza virus (AIV) subtype H9N2 has been evolving rapidly and vaccine escape variants have been reported to cause circulation of infections and economic losses. In the present study, we developed and evaluated ectodomain of the AIV matrix 2 (M2e) protein as a supplementing antigen for oil-based inactivated H9N2 vaccine to increase resistance against vaccine escape variants. AIV H9N2 M2e antigen was expressed in Escherichia coli and supplemented to inactivated H9N2 oil emulsion vaccine. Specific pathogen-free chickens received a single injection of inactivated H9N2 oil emulsion vaccines with or without M2e supplementation. At three weeks post vaccination, hemagglutination inhibition tests and enzyme-linked immunosorbent assays were performed to determine serological immune responses. Challenge study using a vaccine escape H9N2 variant was performed to evaluate the efficacy of M2e supplementation. M2e antigen supplemented in oil emulsion vaccine was highly immunogenic, and a single M2e-supplemented vaccination reduced challenge virus replication and shedding more effectively than non-supplemented vaccination.

Genus Lecithocera Herrich-Schaffer in Vietnam (Lepidoptera, Lecithoceridae, Lecithocerinae), with descriptions of eight new species

Park, K.T.,Kim, S.,Kim, M.Y.,Bae, Y.S. 한국응용곤충학회 2016 Journal of Asia-Pacific Entomology Vol.19 No.2

<P>The Vietnamese species belonging to the genus Lecithocera Herrich-Schaffer of the family Lecithoceridae (Gelechioidea) are reviewed. Thirteen species of Lecithocera including eight new species: Lecithocera alpina Park, sp. nov., L. canitia Park, sp. nov., L. criniptera Park, sp. nov., L. hana Park, sp. nov., L. haviensis Park, sp. nov., L. nara Park, sp. nov., L. orbicuculla Park, sp. nov., and L. tamdaoica Park, sp. nov., are recognized. In the revision, the genus Lecithocera is divided into three species-groups based on the different condition of wing venation: the cuspidata species-group with M-1 and CuA(1) stalked in the hindwing; the rotundata species-group with M-1 and CuA(1) coincident in the hindwing; and the indigens species group with M-2 and M-3 stalked in the forewing. For these newly known species in Vietnam, adults and their genitalia are illustrated, and all available taxonomic data are provided. (C) 2016 Korean Society of Applied Entomology, Taiwan Entomological Society and Malaysian Plant Protection Society. Published by Elsevier B.V. All rights reserved.</P>

닭의 동품종내 계통간 잡종강세 이용시험

박상문,김동곤,송기덕,오봉국 한국축산학회 1965 한국축산학회지 Vol.7 No.1

To induce hybrid vigor by the reciprocal crossing with SWL (Sung Whan Line), MWL (Minnesota Line), AWL (Dembro Line) and BWL (Derby Line) of single comb white leghorn, 1,330 hens were divided into 16 blacks (Diallal cross with four Line) and fed under condition of N.R.C. feeding standard. In this experiment, hatchability, fertility, mortality, body weight, days required on up to first egg laying date, egg weight, egg quality, winter pauses, intensity, feed utilization, number of eggs layed during the testing period of days and brooding were investigated. Cross breeds showed a little higher fertility and hatchability but there were no significance when their parents had high hatchability. Cross breeds M♀×A♂, M♀×E♂ and A♀×S♂ showed more than ½ decrease in mortality, however cross bred hen showed 2.27% more decrease in mortality than purebreeds. Cross breeds of 6 blocks among 16 blocks at 6 weeks showed significant (P$lt;0.01) difference in body weight. Heaviest cross breed among all blocks were S♀×A♂ and A♀×S♂. Generally adult cross breeds showed heaviest body weight; especially in S♀×A♂ and A♀×S♂ cross breeds showed heaviest body weight among them. Dates requiring on up to first egg laying (50% laying date in the blocks) were showen at cross breeds. Cross breeds shorten 16.42 days than purebreeds and A♀×M♂ was showed shortest clay (171.0 days) in all cross breeds. ♀M×A♂ (175.0 days) and M♀×B♂ (175.0 days) were excellent. Cross breed showed increased egg weight as follows : A♀×S♂ 56.0 g, B♀×M♂ 56.62 g B♀×A♂ 57.40 g and there was significant (P$lt;0.05) difference in egg weight between pure breed and cross breed (M♀×S♂ and B♀×A♂), However, their egg weight was same one as standard (56.0 g) Generally productive block showed light egg weight, because egg weight are related to egg production, Those eggs didn't show any progress in thickness, meat and blood spot. Winter pauses became more short and intensity was more higher than purebreeds. They produced more eggs during winter than purebreeds. Feed utilization was very high in cross breeds. than in purebreeds. The number of egg at 500 days testing period (Hen house) was M♀×B♂ : 201,08, S♀×B♂ : 200.73, B♀×S♂ : 200.21, M♀×S♂ : 197.28 and M♀×A♂ : 191.69 and number of egg per hen in the block was from 230 to 256 in a year. SWL and BWL cross was very excellent in any condition. Generally, we could say that the number of egg was increased by crossing and it showed at 1% level of significance, but cross breeds increased broodiness about 0.56% than did in pure breeds.

Aerosol size distributions observed at Naiman in the Asian dust source region of Inner Mongolia

Park, S.U.,Park, M.S. Pergamon Press ; Elsevier [distribution] 2014 Atmospheric environment Vol.82 No.-

Aerosol size distributions of observed mass concentrations at the Naiman site in Inner Mongolia that is one of the major Asian dust source regions have been examined for the period from April 2010 to July 2012. The total number of 262 sampled data using the 10-stage quartz crystal microbalance (QCM) cascade impactor is obtained by presetting the frequency changes of 40 Hz during April 2010, 60 Hz for the period of 28 April-16 September 2010 and 70 Hz from 1 November 2010-29 July 2012. The total mass concentrations (PM<SUB>10</SUB>) measured by the QCM cascade impactor are modified to have the same sampling time of 60 min with the help of the 1-h averaged PM<SUB>10</SUB> concentration measured by the beta gauge at the same site. These modified QCM data are classified into the local dust emission case of 196 and the dust advection case of 66. The local dust emission case is defined when the calculated dust flux with the two-level (3 m and 15 m high) measured PM<SUB>10</SUB> concentrations by the beta-gauge is upward and the PM<SUB>10</SUB> concentration measured at 3 m high exceeds 100 μg m<SUP>-3</SUP> while all the rest of QCM sampled data are classified as the dust advection case. The results indicate that the spectral mass concentration distribution of the local dust emission case shows a two-modal distribution with one additional mode of the large particle that cannot be resolved by the QCM cascade impactor whereas that of the advection case reveals a three-modal distribution with one additional unresolved large particle mode. The percent spectral mass concentration distribution of the unresolved mode (stage 1) for the local dust emission case is larger than that for the dust advection case. The modal distributions of both cases can be regressed optimally with log-normal distribution functions. The resolved log-normal distribution functions of the mass concentration distribution by the QCM cascade impactor are found to be the particle mean diameter (the standard deviation) of 0.28 (2.07) and 3.15 μm (1.41 μm) for the local emission case and 0.16 (1.51), 0.60 (1.41) and 2.88 μm (1.38 μm) for the advection case. This clearly suggests that the spectral mass concentration shifts toward the larger particle size for the local emission case.

한외여과된 돈혈청으로부터 DEAE - Sephacel Chromatography 에 의한 면역단백질의 분리

박우문,유익종,전기홍,이무하 한국축산학회 1997 한국축산학회지 Vol.39 No.1

Concentrated porcine serum with ultrafiltration 30KDa was purified with DEAF-Sephacel column chromatography. Buffer solution with pH 7 to 9 and buffer concentration with 0.01M to 0.05M, and NaCl gradient with 0M∼0.5M, 0M∼0.8M and 0M∼1.0M were used respectively for chromatographic purification. Optimal conditions were at pH 8.0, buffer conc. 0.03M and salt conc, 0M∼0.5M with immunoprotein concentration 82% and production yield 43%. Purified immunoprotein was identified in the molecular weight 50KDa and 15KDa by electrophoresis.

Differential growth of U and M type infectious haematopoietic necrosis virus in a rainbow trout–derived cell line, RTG-2

Park, J W,Moon, C H,Wargo, A R,Purcell, M K,Kurath, G Blackwell Publishing Ltd 2010 Journal of fish diseases Vol.33 No.7

<P>Abstract</P><P>Infectious haematopoietic necrosis virus (IHNV) is one of the most important viral pathogens of salmonids. In rainbow trout, IHNV isolates in the M genogroup are highly pathogenic, while U genogroup isolates are significantly less pathogenic. We show here that, at a multiplicity of infection (MOI) of 1, a representative U type strain yielded 42-fold less infectious virus than an M type strain in the rainbow trout–derived RTG-2 cell line at 24 h post-infection (p.i.). However, at an MOI of 10, there was only fivefold difference in the yield of infectious virus between the U and M strains. Quantification of extracellular viral genomic RNA suggested that the number of virus particles released from cells infected with the U strain at a MOI of 1 was 47-fold lower than from M-infected cells, but U and M virions were equally infectious by particle to infectivity ratios. At an MOI of 1, U strain intracellular viral genome accumulation and transcription were 37- and 12-fold lower, respectively, than those of the M strain at 24 h p.i. Viral nucleocapsid (N) protein accumulation in U strain infections was fivefold lower than in M strain infections. These results suggest that the block in U type strain growth in RTG-2 cells was because of the effects of reduced genome replication and transcription. The reduced growth of the U strain does not seem to be caused by defective genes, because the U and M strains grew equally well in the permissive <I>epithelioma papulosum cyprini</I> cell line at an MOI of 1. This suggests that host-specific factors in RTG-2 cells control the growth of the IHNV U and M strains differently, leading to growth restriction of the U type virus during the RNA synthesis step.</P>