Rat H - Y 항체에 의한 생쥐 Embryo 의 성 조절에 관한 연구 Ⅲ. H - Y 항혈청에 의한 BALB c 생쥐 상실배의 성 판별

정장용(J . Y . Jeung),박충생(C . S . Park),박희성(H . S . Park) 한국축산학회 1989 한국축산학회지 Vol.31 No.10

This experiment was carried out to develop a new technique by identifying XX-bearing embryos prior to implantation by immunological method. H-Y antiserum was prepared in inbred Wistar female rats by repeated immunization with spleen cells from males of the same strain. The reactivity of H-Y antibody was confirmed by culturing mouse embryos in the medium containing H-Y antiserum and complement obtained from the guinea pig. The optimal condition for the activity of H-Y antibody was also investigated by culturing embryos under the various conditions of equilibration times, complement concentrations and various media. The results obtained in this experiment are as follows: When the embryos were cultured in the medium of H-Y antiserum and complement which was given the equilibration time of less than 30 minutes in CO₂ incubator, the lysis-rate of embryo was 89.3%. The embryo lysis-rates in the equilibration time of 1-1.5, 3-3.5, or 24-26 hours were 48.1, 47.7 and 48.2%, respectively. When the concentration rate of complement to H-Y antiserum varied from 0.25 - 4.0, the lysis-rate of embryo was 43.2 to 52.7%. The concentration rate of complement did not influence the lysis-rate of embryos. The meda of D-PBS + 0.3% BSA, D-PBS + 20% FCS, Ham`s F-10 + 0.3% BSA and Ham`s F-10 + 20% FCS showed the embryo lysis-rate of 46.4, 57.4, 49.3 and 49.1%, respectively. The culture media used in this experiment did not show any significant difference in the embryo lysis-rate. After the embryos were cultured to the late blastocyst in the media of D-PBS + NGPS + H-Y antiserum or D-PBS + NGPS + normal female rat serum the normally developed embryos were selected and transferred to the pseudo pregnant recipients. The percentages of their female offspring were 82.3%(14/17) in H-Y antiserum treatment and 53.6%(15/28) in normal serum treatment and showed a significant difference between the two treatments(p $lt;0.001).

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.

Analysis of 22 Y chromosomal STR haplotypes and Y haplogroup distribution in Pathans of Pakistan

Lee, E.Y.,Shin, K.J.,Rakha, A.,Sim, J.E.,Park, M.J.,Kim, N.Y.,Yang, W.I.,Lee, H.Y. Elsevier Science 2014 FORENSIC SCIENCE INTERNATIONAL GENETICS Vol.11 No.-

We analyzed haplotypes for 22 Y chromosomal STRs (Y-STRs), including 17 Yfiler loci (DYS19, DYS385a/b, DYS389I/II, DYS390, DYS391, DYS392, DYS393, DYS437, DY438, DYS439, DYS448, DYS456, DYS458, DYS635 and Y-GATA-H4) and five additional STRs (DYS388, DYS446, DYS447, DYS449 and DYS464), and Y chromosomal haplogroup distribution in 270 unrelated individuals from the Pathans residing in the Federally Administered Tribal Areas and the North-West Frontier Province of Pakistan using in-house multiplex PCR systems. Each Y-STR showed diversities ranging from 0.2506 to 0.8538, and the discriminatory capacity (DC) was 73.7% with 199 observed haplotypes using 17 Yfiler loci. By the addition of 5 Y-STRs to the Yfiler system, the DC was increased to 85.2% while showing 230 observed haplotypes. Among the additional 5 Y-STRs, DYS446, DYS447 and DYS449 were major contributors to enhancing discrimination. In the analysis of molecular variance, the Pathans of this study showed significant differences from other Pathan populations as well as neighboring population sets. In Y-SNP analysis, a total of 12 Y chromosomal haplogroups were observed and the most frequent haplogroup was R1a1a with 49.3% frequency. To obtain insights on the origin of Pathans, the network analysis was performed for the haplogroups G and Q observed from the Pathans and the Jewish population groups including Ashkenazim and Sephardim, but little support for a Jewish origin could be found. In the present study, we report Y-STR population data in Pathans of Pakistan, and we emphasize the need for adding additional markers to the commonly used 17 Yfiler loci to achieve more improved discriminatory capacity in a population with low genetic diversity.

Peroxiredoxin II promotes hepatic tumorigenesis through cooperation with Ras/Forkhead box M1 signaling pathway

Park, Y-H,Kim, S-U,Kwon, T-H,Kim, J-M,Song, I-S,Shin, H-J,Lee, B-K,Bang, D-H,Lee, S-J,Lee, D-S,Chang, K-T,Kim, B-Y,Yu, D-Y Macmillan Publishers Limited 2016 Oncogene Vol.35 No.27

<P>The current study was carried out to define the involvement of Peroxiredoxin (Prx) II in progression of hepatocellular carcinoma (HCC) and the underlying molecular mechanism(s). Expression and function of Prx II in HCC was determined using H-ras(G12V)-transformed HCC cells (H-ras(G12V)-HCC cells) and the tumor livers from H-ras(G12V)-transgenic (Tg) mice and HCC patients. Prx II was upregulated in H-ras(G12V)-HCC cells and H-ras(G12V)-Tg mouse tumor livers, the expression pattern of which highly similar to that of forkhead Box M1 (FoxM1). Moreover, either knockdown of FoxM1 or site-directed mutagenesis of FoxM1-binding site of Prx II promoter significantly reduced Prx II levels in H-ras(G12V)-HCC cells, indicating FoxM1 as a direct transcription factor of Prx II in HCC. Interestingly, the null mutation of Prx II markedly decreased the number and size of tumors in H-ras(G12V)-Tg livers. Consistent with this, knockdown of Prx II in H-ras(G12V)-HCC cells reduced the expression of cyclin D1, cell proliferation, anchorage-independent growth and tumor formation in athymic nude mice, whereas overexpression of Prx II increased or aggravated the tumor phenotypes. Importantly, the expression of Prx II was correlated with that of FoxM1 in HCC patients. The activation of extracellular signal-related kinase (ERK) pathway and the expression of FoxM1 and cyclin D1 were highly dependent on Prx II in H-ras(G12V)-HCC cells and H-ras(G12V)-Tg livers. Prx II is FoxM1-dependently- expressed antioxidant in HCC and function as an enhancer of Ras(G12V) oncogenic potential in hepatic tumorigenesis through activation of ERK/FoxM1/cyclin D1 cascade.</P>

Neural stem cells injured by oxidative stress can be rejuvenated by GV1001, a novel peptide, through scavenging free radicals and enhancing survival signals

Park, H.H.,Yu, H.J.,Kim, S.,Kim, G.,Choi, N.Y.,Lee, E.H.,Lee, Y.J.,Yoon, M.Y.,Lee, K.Y.,Koh, S.H. Elsevier BV 2016 NeuroToxicology Vol.55 No.-

<P>Oxidative stress is a well-known pathogenic mechanism of a diverse array of neurological diseases, and thus, numerous studies have attempted to identify antioxidants that prevent neuronal cell death. GV1001 is a 16-amino-acid peptide derived from human telomerase reverse transcriptase (hTERT). Considering that hTERT has a strong antioxidant effect, whether GV1001 also has an antioxidant effect is a question of interest. In the present study, we aimed to investigate the effects of GV1001 against oxidative stress in neural stem cells (NSCs). Primary culture NSCs were treated with different concentrations of GV1001 and/or hydrogen peroxide (H2O2) for various time durations. The H2O2 decreased the viability of the NSCs in a concentration-dependent manner, with 200 mu M H2O2 significantly decreasing both proliferation and migration. However, treatment with GV1001 rescued the viability, proliferation and migration of H2O2 injured NSCs. Consistently, free radical levels were increased in rat NSCs treated with H2O2, while co-treatment with GV1001 significantly reduced these levels, especially the intracellular levels. In addition, GV1001 restored the expression of survival-related proteins and reduced the expression of death associated ones in NSCs treated with H2O2. In conclusion, GV1001 has antioxidant and neuroprotective effects in NSCs following treatment with H2O2, which appear to be mediated by scavenging free radicals, increasing survival signals and decreasing death signals. (C) 2016 Elsevier B.V. All rights reserved.</P>

Cross-protective efficacies of highly-pathogenic avian influenza H5N1 vaccines against a recent H5N8 virus

Park, S.J.,Si, Y.J.,Kim, J.,Song, M.S.,Kim, S.m.,Kim, E.H.,Kwon, H.i.,Kim, Y.I.,Lee, O.J.,Shin, O.S.,Kim, C.J.,Shin, E.C.,Choi, Y.K. Academic Press 2016 Virology Vol.498 No.-

<P>To investigate cross-protective vaccine efficacy of highly-pathogenic avian influenza H5N1 viruses against a recent HPAI H5N8 virus, we immunized C57BL/6 mice and ferrets with three alum-adjuvanted inactivated whole H5N1 vaccines developed through reverse-genetics (Rg): [Vietnam/1194/04xPR8 (clade 1), Korea/W149/06xPR8 (clade 2.2), and Korea/ES223N/03xPR8 (clade 2.5)]. Although relatively low cross-reactivities (10-40 HI titer) were observed against heterologous H5N8 virus, immunized animals were 100% protected from challenge with the 20 mLD(50) of H5N8 virus, with the exception of mice vaccinated with 3.5 mu g of Rg Vietnam/1194/04xPR8. Of note, the Rg Korea/ES223N/03xPR8 vaccine provided not only effective protection, but also markedly inhibited viral replication in the lungs and nasal swabs of vaccine recipients within five days of HPAI H5N8 virus challenge. Further, we demonstrated that antibody-dependent cell-mediated cytotoxicity (ADCC) of an antibody-coated target cell by cytotoxic effector cells also plays a role in the heterologous protection of H5N1 vaccines against H5N8 challenge. (C) 2016 Elsevier Inc. All rights reserved.</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.

Effects of HA and NA glycosylation pattern changes on the transmission of avian influenza A(H7N9) virus in guinea pigs

Park, S.,Lee, I.,Kim, J.I.,Bae, J.Y.,Yoo, K.,Kim, J.,Nam, M.,Park, M.,Yun, S.H.,Cho, W.I.,Kim, Y.S.,Ko, Y.Y.,Park, M.S. Academic Press 2016 Biochemical and biophysical research communication Vol.479 No.2

Avian influenza H7N9 virus has posed a concern of potential human-to-human transmission by resulting in seasonal virus-like human infection cases. To address the issue of sustained human infection with the H7N9 virus, here we investigated the effects of hemagglutinin (HA) and neuraminidase (NA) N-linked glycosylation (NLG) patterns on influenza virus transmission in a guinea pig model. Based on the NLG signatures identified in the HA and NA genetic sequences of H7N9 viruses, we generated NLG mutant viruses using either HA or NA gene of a H7N9 virus, A/Anhui/0½013, by reverse genetics on the 2009 pandemic H1N1 virus backbone. For the H7 HA NLG mutant viruses, NLG pattern changes appeared to reduce viral transmissibility in guinea pigs. Intriguingly, however, the NLG changes in the N9 NA protein, such as a removal from residue 42 or 66 or an addition at residue 266, increased transmissibility of the mutant viruses by more than 33%, 50%, and 16%, respectively, compared with a parental N9 virus. Given the effects of HA-NA NLG changes with regard to viral transmission, we then generated the HA-NA NLG mutant viruses harboring the H7 HA of double NLG addition and the N9 NA of various NLG patterns. As seen in the HA NLG mutants above, the double NLG-added H7 HA decreased viral transmissibility. However, when the NA NLG changes occurred by a removal of residue 66 and an addition at 266 were additionally accompanied, the HA-NA NLG mutant virus recovered the transmissibility of its parental virus. These demonstrate the effects of specific HA-NA NLG changes on the H7N9 virus transmission by highlighting the importance of a HA-NA functional balance.

Effects of Fertilization Time and Culture Medium of Pig Oocytes Matured In Vitro by Liquid Boar Sperm Stored at 4℃

Park, C.S.,Yi, Y.J.,Kim, M.Y.,Chang, Y.J.,Lee, S.H.,Jin, D.I 충남대학교 형질전환복제돼지연구센터 2004 논문집 Vol. No.8

This study was to investigate the effects of fertilization time and culture medium of pig oocytes matured in-vitro by liquid boar sperm. The sperm rich fraction (30∼60 ml) was slowly cooled to room temperature (20∼23℃) by 2 h after collection. Semen was transferred into 15 ml tubes, centrifuged at room temperature for 10 min 800×g, and the supernatant solution was poured off. The concentrated sperm was resuspended with 5 ml of the LEN diluent to provide 1.0×10^(9) sperm/ml at room temperature. The resuspended semen was cooled in a refrigerator to 4℃. The medium used for oocyte maturation was TCM-199 supplemented with 26.19 mM sodium bicarbonate, 0.9 mM sodium pyruvate, 10μg/ml insulin, 2μg/ml vitamin B_(12), 25 mM HEPES, 10μg/ml bovine apotransferrin, 150μM cysteamine, 10IU/ml PMSG, 10 IU/ml hCG, 10 ng/ml EGF, 0.4% BSA, 75μg/ml sodium penicillin G, 50μg/ml streptomycin sulfate and 10% pFF. After about 22 h of culture, oocytes were cultured without cysteamine and hormones for 22 h at 38.5℃, 5% CO₂in air. Oocytes were inseminated with liquid boar sperm stored at 4℃ for 2 days after collection. Oocytes were coincubated for 1, 3, 6 and 9 h in 500 μl mTBM fertilization media with 1.0×10^(6) sperm/ml concentration, respectively. Thereafter, oocytes were transferred into 500 μl NCSU-23, HEPES buffered NCSU-23, PZM-3 and PZM-4 culture media, respectively, for further culture of 6, 48 and 144 h. The rates of sperm penetration and male pronuclear formation were higher in the fertilization times for 6 and 9 h than in those for 1 and 3 h. The rates of cleaved oocytes were higher in the fertilization times for 6 and 9 h (85.0 and 84.6%) than in those for 1 and 3 h (61.1 and 76.8%). The percentage of blastocyst formation from the cleaved oocytes was highest in the fertilization time for 6 h (33.6%) than in that for 1, 3 and 9 h (11.4, 23.0 and 29.6%). Mean cell numbers per blastocyst were 32.9, 27.6, 26.3 and 24.4 in the fertilization times for 6, 9, 3 and 1 h, respectively. The rate of blastocyst from the cleaved oocytes and the number of cells per blastocyst were higher in HEPES buffered NCSU-23 culture medium than in NCSU-23, PZM-3 and PZM-4 culture media. In conclusion, we found out that liquid boar sperm stored at 4℃ could be used for in-vitro fertilization of pig oocytes matured in-vitro. Also, we recommend the coincubation time of 6 h in 500 μl TBM fertilization medium with 1×10^(6) sperm/ml concentration and the HEPES buffered NCSU-23 culture medium for in-vitro fertilization of pig oocytes matured in-vitro.

Effects of calcium doping on the superconducting properties of top-seeded melt growth processed Y_1.5Ba_2-xCa_xCu₃O_y superconductors

Park, S.D.,Kim, H.J.,Park, B.J.,Han, Y.H.,Jun, B.H.,Lee, J.S.,Kim, C.J. North-Holland 2011 Physica. C, Superconductivity Vol.471 No.21

The effect of calcium doping on the superconducting properties of top seeded melt growth (TSMG) processed Y<SUB>1.5</SUB>Ba<SUB>2-x</SUB>Ca<SUB>x</SUB>Cu<SUB>3</SUB>O<SUB>y</SUB> superconductors was studied in terms of calcium content (X<SUB>ca</SUB>). YBa<SUB>2-x</SUB>Ca<SUB>x</SUB>Cu<SUB>3</SUB>O<SUB>7-δ</SUB> (X<SUB>ca</SUB>=0, 0.005, 0.01, 0.02, 0.04, 0.1, 0.3) powders were synthesized by the powder calcination method. YBa<SUB>2-x</SUB>Ca<SUB>x</SUB>Cu<SUB>3</SUB>O<SUB>7-δ</SUB> powders were mixed with 0.25mole Y<SUB>2</SUB>O<SUB>3</SUB> powder and 1wt.% CeO<SUB>2</SUB> as Y<SUB>2</SUB>BaCuO<SUB>5</SUB> (Y211) refiner, and finally made into Y<SUB>1</SUB>.<SUB>5</SUB>Ba<SUB>2-x</SUB>Ca<SUB>x</SUB>Cu<SUB>3</SUB>O<SUB>y</SUB> (Y1.5)+1wt.% CeO<SUB>2</SUB> composition. The single Y123 growth on the top surface was observed up to X<SUB>ca</SUB>=0.1, while the multiple Y123 growth was observed at X<SUB>ca</SUB>≥0.1. The superconducting transition temperature (T<SUB>c</SUB>) and critical current density (J<SUB>c</SUB>) of TSMG processed Y1.5 samples were inversely proportional to X<SUB>ca</SUB>. The Y211 size increased with increasing X<SUB>ca</SUB> due to the enhancement of Y211 coarsening by calcium doping. No Y211 refining effect by CeO<SUB>2</SUB> was observed in the calcium doped samples. The T<SUB>c</SUB> and J<SUB>c</SUB> decrease by calcium doping are likely to be due to the calcium incorporation with the Y123 lattice and formation of coarse Y211 particles.