H-Y 에 대한 단일클론 항체의 생산 및 그 이용에 관한 연구 1 . H-Y 에 대한 단일클론항체의 생산

심호섭(H . S . Shim),김재화(J . H . Kim),이병철(B . C . Lee),김종배(J . B . Kim),박홍양(H . Y . Park),정길생(K . S . Chung) 한국축산학회 1988 한국축산학회지 Vol.30 No.7

Testis supernatant, a source of H-Y, obtained from BALB/c mice was used to immunize females of same strain. B lymphocytes of mouse producing antibodies to H-Y were fused with SP2/0-Ag 14 myeloma cells and distributed to 384 wells of 96-well microtiter plates. Eighty hybridoma colonies were formed, resulting in 20.8 percent of fusion efficiency. Three strong positive wells from hybridoma colonies were selected for cloning by ELISA and two of them were also found to be positive by indirect immunofluorescence test. Twelve wells of ELISA-positive were selected after cloning and 2D45D4 clones from them were confirmed to produce monoclonal antibodies to H-Y by indirect immunofluorescence test.

受精能獲得精子와 山羊卵子의 體外受精에 관한 硏究

송해범,심금섭 대구대학교 농업과학연구소 1987 農業科學硏究論文集 Vol.1 No.-

Ejaculated and/or epididymal spermatozoa of goats were preincubated for 4-6 h in the isolated genital tracts from hamsters, gilts and goats, or for 3-8 h in m-KRB solution. After preincubation, they inseminated the goat ovulated eggs collected shortly after ovulation and/or the follicular oocytes with intact and without cumulus cells cultured for 25 h in m-KRB solution, and the oocytes with dispersed cumulus cells which were not cultured. The results obtained were as follows. (1) None of the ovulated eggs were fertilized after insemination with ejaculated and epididymal spermatozoa preincubated for 4-5 h in the isolated genital tracts from hamsters, gilts and goats. But 50 and 38% of the ovulated eggs were fertilized with epididymal sermatozoa preincubated for 5 and 6 h in m-KRB solution, respectively. (2) None of the follicular oocytes were fertilized after insemination with ejaculated spermatozoa preincubated for 4-6 h in the uteri isolated from hamsters and goats, and with epididymal spermatozoa preincubated for 4-6 h in the isolated hamster uterus. But 38% of the follicular oocytes were fertilized with epididymal spermatozoa preincubated for 5 h in the isolated goat uterus. (3) After further culture with spermatozoa for 18-24 h, 50-89, 89 and 14-33% of the oocytes with intact, with dispersed and without cumulus cells had matured to the second metaphase, respectively. (4) When epididymal spermatozoa were preincubated for 5 h at the concentration of 4.2x10^(8)/ml and 6 h at the concentration of 3.5x10^(8)/ml in m-KRB solution, 36 and 33% of the oocytes with intact cumulus cells were fertilized. (5) The results suggested that epididymal spermatozoa can be capacitated in m-KRB solution, and that the follicular oocytes matured in culture could be used for the study of fertilization in vitro instead of the ovulated eggs.

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.

생쥐수정란에 대한 H - Y 항체처리가 산자의 성비에 미치는 영향

심호섭,고정재,김종배,박홍양,정길생 ( H . S . Shim,J . J . Ko,J . B . Kim,H . Y . Park,K . S . Chung ) 한국축산학회 1986 한국축산학회지 Vol.28 No.12

These experiments were carried out to control the sex of offsprings in mice by sexing embryos using immunological means prior to transfer to pseudopregnant recipients. H-Y antisera were prepared in inbred SD female rats by repeated immunization of testis supernatant and spleen cells from males of same strain. ELISA and indirect immunofluorescence test were used to detect H-Y antibody in antisera. Eight- to 16-cell mouse embryos were cultured in medium with H-Y antibody and complement (treated embryos) and in medium with BSA (control embryos). After 24-48 hr of culture, embryos were observed their morphological characteristics under the phase contrast microscope. Embryos developed to normal blastocyst were transferred to pseudopregnant recipients and sex of resultant offspring was investigated. The results obtained in these experiments were summarized as follows: 1. Production of H-Y antibodies in antisera obtained from immunized rats was confimed by ELISA and indirect immunofluorescence test. 2. Of 270 embryos treated with H-Y antibody and complement, 126 embryos (46.7%) were developed to normal blastocysts. 3. Following transfer of 126 blastocysts, 16 embryos (12.6%) were survived to term and 13 females (81.3%), significantly high ratio of female offspring, were produced.

Pharmacokinetics and milk distribution characteristics of orbifloxacin following intravenous and intramuscular injection in lactating ewes

GOUDAH, A.,CHO, H.-J.,SHIN, H.-C.,SHIM, J.-H.,REGMI, N. L.,SHIMODA, M.,ABD EL-ATY, A. M. Blackwell Publishing Ltd 2009 JOURNAL OF VETERINARY PHARMACOLOGY AND THERAPEUTIC Vol.32 No.4

<P>The purpose of the current investigation is to elucidate the pharmacokinetic profiles of orbifloxacin (OBFX) in lactating ewes (<I>n </I>= 6) following intravenous (i.v.) and intramuscular (i.m.) administrations of 2.5 mg/kg W. In a crossover study, frequent blood, milk, and urine samples were drawn for up to 48 h after the end of administration, and were then assayed to determine their respective drug concentrations through microbiological assay using <I>Klebsiella pneumoniae</I> as the test micro-organism. Plasma pharmacokinetic parameters were derived from plasma concentration–time data using a compartmental and noncompartmental analysis, and validated a relatively rapid elimination from the blood compartment, with a slope of the terminal phase of 0.21 ± 0.02 and 0.19 ± 0.06 per hour and a half-life of 3.16 ± 0.43 and 3.84 ± 0.59 h, for i.v. and i.m. dosing, respectively. OBFX was widely distributed with a volume of distribution <I>V</I>(<SUB>d(ss)</SUB>) of 1.31 ± 0.12 L/kg, as suggested by the low percentage of protein binding (22.5%). The systemic body clearance (<I>Cl</I><SUB>B</SUB>) was 0.32 ± 0.12 L/h·kg. Following i.m. administration, the maximum plasma concentration (<I>C</I><SUB>max</SUB>) of 1.53 ± 0.34 &mgr;g/mL was reached at <I>t</I><SUB>max</SUB> 1.25 ± 0.21 h. The drug was completely absorbed after i.m. administration, with a bioavailability of 114.63 ± 11.39%. The kinetic milk <I>AUC</I><SUB>milk</SUB>/<I>AUC</I><SUB>plasma</SUB> ratio indicated a wide penetration of orbifloxacin from the bloodstream to the mammary gland. OBFX urine concentrations were higher than the concurrent plasma concentrations, and were detected up to 30 h postinjection by both routes. Taken together, these findings indicate that systemic administration of orbifloxacin could be efficacious against susceptible mammary and urinary pathogens in lactating ewes.</P>

Effect of substrate temperature on the nanostructural and chemical features of nc-Si:H thin films prepared by PECVD

Son, J.I.,Shim, J.H.,Cho, N.H. Elsevier 2010 CURRENT APPLIED PHYSICS Vol.10 No.3

Hydrogenated nanocrystalline silicon (nc-Si:H) thin films were deposited by plasma enhanced chemical vapor deposition (PECVD) at a RF power of 100W; SiH<SUB>4</SUB> and H<SUB>2</SUB> were introduced into a reaction chamber at 25 and 40sccm, respectively, and the substrate temperature ranged from room temperature to 600<SUP>o</SUP>C. The effect of the substrate temperature on the formation of nanoscale Si crystallites (nc-Si) and their structural and optical features were investigated. The average size and concentration of the Si nanocrystallites varied with the substrate temperature; the former ranged from ∼1.0 to ∼5.0nm, and the latter reached up to ∼15.5% when the substrate temperature was 400<SUP>o</SUP>C. With increasing substrate temperature to 400<SUP>o</SUP>C, the relative fraction of Si-H bonds in the films, [Si-H]/@?<SUB>n=1</SUB><SUP>3</SUP>[Si-H<SUB>n</SUB>]<SUB>n=integer</SUB>, was increased up to ∼29.3%.

Pharmacokinetics and Mammary Residual Depletion of Erythromycin in Healthy Lactating Ewes

Goudah, A.,Sher Shah, S.,Shin, H.C.,Shim, J.H.,Abd El-Aty, A. M. Blackwell Publishing Ltd 2007 Journal of veterinary medicine. A, Physiology, pat Vol.54 No.10

<P>Summary</P><P>The aim of this investigation was to examine the pharmacokinetics and mammary excretion of erythromycin administered to lactating ewes (<I>n</I> = 6) by the intravenous (i.v.), intramuscular (i.m.) and subcutaneous (s.c.) routes at a dosage of 10 mg/kg. Blood and milk samples were collected at pre-determined times, and a microbiological assay method was used to measure erythromycin concentrations in serum and milk. The concentration–time data were analysed by compartmental and non-compartmental kinetic methods. The serum concentration–time data of erythromycin were fit to a two-compartment model after i.v. administration and a one-compartment model with first-order absorption after i.m. and s.c. administration. The elimination half-life (<I>t</I><SUB>1/2&bgr;</SUB>) was 4.502 ± 1.487 h after i.v. administration, 4.874 ± 0.296 h after i.m. administration and 6.536 ± 0.151 h after s.c. administration. The clearance value (Cl<SUB>tot</SUB>) after i.v. dosing was 1.292 ± 0.121 l/h/kg. After i.m. and s.c. administration, observed peak erthyromycin concentrations (<I>C</I><SUB>max</SUB>) of 0.918 ± 0.092 <I>&mgr;</I>g/ml and 0.787 ± 0.010 <I>&mgr;</I>g/ml were achieved at 0.75 and 1.0 h (<I>T</I><SUB>max</SUB>) respectively. The bioavailability obtained after i.m. and s.c. administration was 91.178 ± 10.232% and 104.573 ± 9.028% respectively. Erythromycin penetration from blood to milk was quick for all the routes of administration, and the high AUC<SUB>milk</SUB>/AUC<SUB>serum</SUB> (1.186, 1.057 and 1.108) and C<SUB>max‐milk</SUB>/C<SUB>max‐serum</SUB> ratios reached following i.v., i.m. and s.c. administration, respectively, indicated an extensive penetration of erythromycin into the milk.</P>

Characterization of the pharmacokinetic disposition of levofloxacin in stallions after intravenous and intramuscular administration

GOUDAH, A.,ABO EL-SOOUD, K.,SHIM, J.-H.,SHIN, H.-C.,ABD EL-ATY, A. M. Blackwell Publishing Ltd 2008 JOURNAL OF VETERINARY PHARMACOLOGY AND THERAPEUTIC Vol.31 No.5

<P>The target of the present study was to investigate the plasma disposition kinetics of levofloxacin in stallions (<I>n</I> = 6) following a single intravenous (i.v.) bolus or intramuscular (i.m.) injection at a dose rate of 4 mg/kg bwt, using a two-phase crossover design with 15 days as an interval period. Plasma samples were collected at appropriate times during a 48-h administration interval, and were analyzed using a microbiological assay method. The plasma levofloxacin disposition was best fitted to a two-compartment open model after i.v. dosing. The half-lives of distribution and elimination were 0.21 ± 0.13 and 2.58 ± 0.51 h, respectively. The volume of distribution at steady-state was 0.81 ± 0.26 L/kg, the total body clearance (<I>Cl</I><SUB>tot</SUB>) was 0.21 ± 0.18 L/h/kg, and the areas under the concentration–time curves (<I>AUC</I>s) were 18.79 ± 4.57 &mgr;g.h/mL. Following i.m. administration, the mean <I>t</I><SUB>1/2el</SUB> and <I>AUC</I> values were 2.94 ± 0.78 h and 17.21 ± 4.36 &mgr;g.h/mL. The bioavailability was high (91.76% ± 12.68%), with a peak plasma mean concentration (<I>C</I><SUB>max</SUB>) of 2.85 ± 0.89 &mgr;g/mL attained at 1.56 ± 0.71 h (<I>T</I><SUB>max</SUB>). The <I>in vitro</I> protein binding percentage was 27.84%. Calculation of efficacy predictors showed that levofloxacin might have a good therapeutic profile against Gram-negative and Gram-positive bacteria, with an <I>MIC</I> ≤ 0.1 &mgr;g/mL.</P>

Combined steam and carbon dioxide reforming of methane and side reactions: Thermodynamic equilibrium analysis and experimental application

Jang, W.J.,Jeong, D.W.,Shim, J.O.,Kim, H.M.,Roh, H.S.,Son, I.H.,Lee, S.J. Applied Science Publishers 2016 APPLIED ENERGY Vol.173 No.-

<P>Thermodynamic equilibrium analysis of the combined steam and carbon dioxide reforming of methane (CSCRM) and side reactions was performed using total Gibbs free energy minimization. The effects of (CO2 + H2O)/CH4 ratio (0.9-2.9), CO2:H2O ratio (3:1-1:3), and temperature (500-1000 degrees C) on the equilibrium conversions, yields, coke yield, and H-2/CO ratio were investigated. A (CO2 + H2O)/CH4 ratio greater than 1.2, a CO2:H2O ratio of 1:2.1, and a temperature of at least 850 degrees C are preferable reaction conditions for the synthesis gas preparation in the gas to liquid process. Simulated conditions were applied to the CSCRM reaction and the experimental data were compared with the thermodynamic equilibrium results. The thermodynamic equilibrium results were mostly consistent with the experimental data, but the reverse water gas shift reaction rapidly occurred in the real chemical reaction and under excess oxidizing agent conditions. In addition, a long-term stability test (under simulated conditions) showed that the equilibrium conversion was maintained for 500 h and that the coke formation on the used catalyst was not observed. (C) 2016 Elsevier Ltd. All rights reserved.</P>

Molecular characterisation and expression analysis of the cathepsin H gene from rock bream (Oplegnathus fasciatus)

Kim, J.W.,Park, C.I.,Hwang, S.D.,Jeong, J.M.,Kim, K.H.,Kim, D.H.,Shim, S.H. Academic Press 2013 Fish & shellfish immunology Vol.35 No.1

Cathepsins are lysosomal cysteine proteases belonging to the papain family, whose members play important roles in normal metabolism for the maintenance of cellular homeostasis. Rock bream (Oplegnathus fasciatus) cathepsin H (RbCTSH) cDNAs were identified by expressed sequence tag analysis of a lipopolysaccharide-stimulated rock bream liver cDNA library. The full-length RbCTSH cDNA (1326 bp) contained an open reading frame of 978 bp encoding 325 amino acids. The presence of an ERFNIN-like motif was predicted in the propeptide region of RbCTSH. Furthermore, multiple alignments showed that the EPQNCSAT region was well conserved among other cathepsin H sequences. Phylogenetic analysis revealed that RbCTSH is most closely related to Nile tilapia cathepsin H. RbCTSH was expressed significantly in the intestine, spleen, head kidney and stomach. RbCTSH mRNA expression was also examined in several tissues under conditions of bacterial and viral challenge. All examined tissues of fish infected with Edwardsiella tarda, Streptococcus iniae and red sea bream iridovirus (RSIV) showed significant increases in RbCTSH expression compared to the control. In the kidney and spleen, RbCTSH mRNA expression was upregulated markedly following infection with bacterial pathogens. These findings indicate that RbCTSH plays an important role in the innate immune response of rock bream. Furthermore, these results provide important information for the identification of other cathepsin H genes in various fish species.