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.

Ex situ catalytic upgrading of lignocellulosic biomass components over vanadium contained H-MCM-41 catalysts

Kim, B.S.,Jeong, C.S.,Kim, J.M.,Park, S.B.,Park, S.H.,Jeon, J.K.,Jung, S.C.,Kim, S.C.,Park, Y.K. Elsevier Science Publishers 2016 CATALYSIS TODAY - Vol.265 No.-

<P>H-V-MCM-41 catalysts containing 5, 10, and 30 wt% of vanadium were synthesized and applied to the ex situ catalytic pyrolysis (CP) of three polymeric components of lignocellulosic biomass for the first time. Characterization of the catalysts was performed using N-2 adsorption-desorption, XRD, FT-IR, and NH3-TPD. The results of XRD analysis showed that 5 wt% and 10 wt% H-V-MCM-41 catalysts maintained the mesoporous structure, whereas the mesoporous structure was destroyed in 30 wt% H-V-MCM-41 with considerable amount of small V2O5 crystalline outside the framework. NH3-TPD showed that H-V-MCM-41 has mostly weak acid sites and that 10 wt% H-V-MCM-41 had the largest quantity of acid sites due to framework vanadium. In the case of CP of cellulose using Py-GC/MS, 10 wt% H-V-MCM-41 showed the highest catalytic activity for the production of valuable furanic compounds such as furfural because of the enhanced deoxygenation over the acid sites formed on framework vanadium. In the case of CP of xylan as well, 10 wt% H-V-MCM-41 led to the largest yield of mono-aromatics. The production of acetic acid was also promoted by H-V-MCM-41 catalysts. The CP of lignin over H-V-MCM-41 catalysts promoted substantially the production of important feedstock chemicals for the petrochemical industry: phenolics and mono-aromatics. (C) 2015 Elsevier B.V. All rights reserved.</P>

TmSR-C, scavenger receptor class C, plays a pivotal role in antifungal and antibacterial immunity in the coleopteran insect Tenebrio molitor

Kim, S.G.,Jo, Y.H.,Seong, J.H.,Park, K.B.,Noh, M.Y.,Cho, J.H.,Ko, H.J.,Kim, C.E.,Tindwa, H.,Patnaik, B.B.,Bang, I.S.,Lee, Y.S.,Han, Y.S. Pergamon Press ; Elsevier Science Ltd 2017 Insect biochemistry and molecular biology Vol.89 No.-

Scavenger receptors (SRs) constitute a family of membrane-bound receptors that bind to multiple ligands. The SR family of proteins is involved in removing cellular debris, oxidized low-density lipoproteins, and pathogens. Specifically, class C scavenger receptors (SR-C) have also been reported to be involved in phagocytosis of gram-positive and -negative bacteria in Drosophila and viruses in shrimp. However, reports are unavailable regarding the role of SR-C in antifungal immune mechanisms in insects. In this study, a full-length Tenebrio molitor SR-C (TmSR-C) sequence was obtained by 5'- and 3'-Rapid amplification of cDNA ends-polymerase chain reaction (RACE-PCR). The TmSR-C full-length cDNA comprised 1671 bp with 5'- and 3'-untranslated regions of 23- and 107-bp, respectively. TmSR-C encodes a putative protein of 556 amino acid residues that is constitutively expressed in all tissues of late instar larvae and 2-day-old adults, with the highest transcript levels observed in hemocytes of larvae and adults. TmSR-C mRNA showed a 2.5-fold and 3-fold increase at 24 and 6 h after infection with Candida albicans and β-glucan, respectively. Immunoassay with TmSR-C polyclonal antibody showed induction of the putative protein in the cytosols of hemocytes at 3 h after inoculation of C. albicans. RNA interference (RNAi)-based gene silencing and phagocytosis assays were used to understand the role of TmSR-C in antifungal immunity. Silencing of TmSR-C transcripts reduced the survivability of late instar larvae at 2 days post-inoculation of C. albicans, Escherichia coli, or Staphylococcus aureus. Furthermore, in TmSR-C-silenced larvae, there was a decline in the rate of microorganism phagocytosis. Taken together, results of this study suggest that TmSR-C plays a pivotal role in phagocytosing not only fungi but also gram-negative and -positive bacteria in T. molitor.

Ion scattering spectroscopy study of the Si(001)c(4x4)-C surface reconstruction

Park J. Y.,Chae K. H.,Choi D. S.,Kim J. Y.,Kim S. S.,Seo J. H.,Whang C. N. 한국물리학회 2004 THE JOURNAL OF THE KOREAN PHYSICAL SOCIETY Vol.45 No.3

Reconstructed Si(001)c(4 4)-C surface has been studied by coaxial impact collision ion scattering spectroscopy (CAICISS). When 100L ethylene (C2H4) was exposed on Si(001)-(2 1) surface at 700 C, Si(001) dimer structures were changed by induced carbon (C) atoms. The experimental CAICISS spectra and simulation results reveal that the reconstructed Si(001)c(4 4)-C surface shows good agreement with the missing dimer model, rather than the Si-C heterodimer model, and adsorbed C atoms in uence only the reconstructed vertical plane of Si(001) surface. On comparing the azimuthal-scan curves for 100L C/Si(001) with those for clean Si(001), it can be suggested that C atoms occupy the fourth subsurface layer of Si(001) directly below the HB (bridge) site. These results are new evidence supporting the previous studies based on the C incorporation into Si(001) surface with missing dimers and the substitution of the fourth Si layers.

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>

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).

Genetic and phylogenetic characterizations of a novel genotype of highly pathogenic avian influenza (HPAI) H5N8 viruses in 2016/2017 in South Korea

Kim, Y.I.,Park, S.J.,Kwon, H.I.,Kim, E.H.,Si, Y.J.,Jeong, J.H.,Lee, I.W.,Nguyen, H.D.,Kwon, J.J.,Choi, W.S.,Song, M.S.,Kim, C.J.,Choi, Y.K. Elsevier Science 2017 INFECTION GENETICS AND EVOLUTION Vol.53 No.-

<P>During the outbreaks of highly pathogenic avian influenza (HPAI) H5N6 viruses in 2016 in South Korea, novel H5N8 viruses were also isolated from migratory birds. Phylogenetic analysis revealed that the HA gene of these H5N8 viruses belonged to clade 2.3.4.4, similarly to recent H5Nx viruses, and originated from A/Brk/Korea/Gochang1/14(H5N8), a minor lineage of H5N8 that appeared in 2014 and then disappeared. At least four reassortment events occurred with different subtypes (H5N8, H7N7, H3N8 and H10N7) and a chicken challenge study revealed that they were classified as HPAI viruses according to OIE criteria. (C) 2017 Elsevier B.V. All rights reserved.</P>

Advanced H2O2 oxidation for diethyl phthalate degradation in treated effluents: effect of nitrate on oxidation and a pilot-scale AOP operation

Ko, K. B.,Park, C. G.,Moon, T. H.,Ahn, Y. H.,Lee, J. K.,Ahn, K. H.,Park, J. H.,Yeom, I. T. IWA Publishing 2008 Water Science & Technology Vol.58 No.5

<P>One of the objectives of this study was to delineate the effect of nitrate on diethyl phthalate (DEP) oxidation by conducting a bench-scale ultraviolet (UV)/H2O2 and O3/H2O2 operations as suggested in a previous study. We also aim to investigate DEP oxidation at various UV doses and H2O2 concentrations by performing a pilot-scale advanced oxidation processes (AOP) system, into which a portion of the effluent from a pilot-scale membrane bioreactor (MBR) plant was pumped. In the bench-scale AOP operation, the O3 oxidation alone as well as the UV irradiation without H2O2 addition could be among the desirable alternatives for the efficient removal of DEP dissolved in aqueous solutions at a low DEP concentration range of 85±15 μg/L. The adverse effect in the UV/H2O2 process was significantly greater than that in the UV oxidation alone, and its oxidation was almost halved by the nitrate. However, the nitrate clearly enhanced the DEP oxidation in the O3 oxidation and O3/H2O2 process. Especially, the addition of nitrate almost doubled the DEP oxidation efficiency in the O3/H2O2 process. The series of pilot-scale AOP operations confirmed that about 30-50% of DEP dissolved in the treated MBR effluent streams was, at least, oxidized by the O3 oxidation alone as well as the UV irradiation without H2O2 addition. The UV photolysis of H2O2 was most effective for DEP degradation with an H2O2 concentration of 40 mg/L at a UV dose of 500 mJ/cm2.</P>

Gut-Specific Delivery of T-Helper 17 Cells Reduces Obesity and Insulin Resistance in Mice

Hong, C.P.,Park, A.,Yang, B.G.,Yun, C.H.,Kwak, M.J.,Lee, G.W.,Kim, J.H.,Jang, M.S.,Lee, E.J.,Jeun, E.J.,You, G.,Kim, K.S.,Choi, Y.,Park, J.H.,Hwang, D.,Im, S.H.,Kim, J.F.,Kim, Y.K.,Seoh, J.Y.,Surh, C. Elsevier North Holland [etc.] 2017 Gastroenterology Vol.152 No.8

<P>BACKGROUND & AIMS: Obesity and metabolic syndrome have been associated with alterations to the intestinal microbiota. However, few studies examined the effects of obesity on the intestinal immune system. We investigated changes in subsets of intestinal CD4(+) T-helper (T-H) cells with obesity and the effects of gut-tropic T(H)17 cells in mice on a high-fat diet (HFD). METHODS: We isolated immune cells from small intestine and adipose tissue of C57BL/6 mice fed a normal chow diet or a HFD for 10 weeks and analyzed the cells by flow cytometry. Mice fed a vitamin A-deficient HFD were compared with mice fed a vitamin A-sufficient HFD. Obese RAG1-deficient mice were given injections of only regulatory T cells or a combination of regulatory T cells and T(H)17 cells (wild type or deficient in integrin beta 7 subunit or interleukin 17 [IL17]). Mice were examined for weight gain, fat mass, fatty liver, glucose tolerance, and insulin resistance. Fecal samples were collected before and after T cell transfer and analyzed for microbiota composition by metagenomic DNA sequencing and quantitative polymerase chain reaction. RESULTS: Mice placed on a HFD became obese, which affected the distribution of small intestinal CD4(+) T-H cells. Intestinal tissues from obese mice had significant reductions in the proportion of T(H)17 cells but increased proportion of T(H)1 cells, compared with intestinal tissues from nonobese mice. Depletion of vitamin A in obese mice further reduced the proportion of T(H)17 cells in small intestine; this reduction correlated with more weight gain and worsening of glucose intolerance and insulin resistance. Adoptive transfer of in vitro-differentiated gut-tropic T(H)17 cells to obese mice reduced these metabolic defects, which required the integrin beta 7 subunit and IL17. Delivery of T(H)17 cells to intestines of mice led to expansion of commensal microbes associated with leanness. CONCLUSIONS: In mice, intestinal T(H)17 cells contribute to development of a microbiota that maintains metabolic homeostasis, via IL17. Gut-homing T(H)17 cells might be used to reduce metabolic disorders in obese individuals.</P>

Preparation of [bis(amido)-phosphine] and [amido-phosphine sulfide or oxide] hafnium and zirconium complexes for olefin polymerization

Lee, C.S.,Park, J.H.,Hwang, E.Y.,Park, G.H.,Go, M.J.,Lee, J.,Lee, B.Y. Elsevier Sequoia 2014 Journal of organometallic chemistry Vol.772 No.-

New phosphine-based bidentate ligands, 2-Me-8-Ph<SUB>2</SUB>P(X)C<SUB>9</SUB>H<SUB>8</SUB>NH (3, X = O; 4, X = S) and N-R-2-Ph<SUB>2</SUB>P(X)C<SUB>6</SUB>H<SUB>4</SUB>NH (5, R = Et, X = O; 6, R = Me, X = S; 7, R = Et, X = S) were prepared via ortho-lithiation of 1,2,3,4-tetrahydroquinaldine (2-Me-C<SUB>9</SUB>H<SUB>9</SUB>NH) and aniline derivatives (N-R-C<SUB>6</SUB>H<SUB>5</SUB>NH). Reaction of the ortho-lithiated compounds with 0.5 equiv of PhP(OPh)<SUB>2</SUB> afforded the bis(amido)-phosphine ligands (2-Me-C<SUB>9</SUB>H<SUB>8</SUB>NH-8-yl)<SUB>2</SUB>PPh (8) and (N-R-C<SUB>6</SUB>H<SUB>4</SUB>NH-2-yl)<SUB>2</SUB>PPh (9, R = Me; 10, R = Et). Using these ligands, [amido-phosphine oxide]Hf(CH<SUB>2</SUB>Ph)<SUB>3</SUB>, [amido-phosphine sulfide]Hf(CH<SUB>2</SUB>Ph)<SUB>3</SUB>, [bis(amido)-phosphine]MX<SUB>2</SUB> (M = Hf, Zr; X = CH<SUB>2</SUB>Ph, Cl, Me), and [amido-phosphine-amine]MCl<SUB>3</SUB> complexes were prepared. The molecular structures of [amido-phosphine sulfide]Hf(CH<SUB>2</SUB>Ph)<SUB>3</SUB> (13) (prepared using 6), [bis(amido)-phosphine]ZrMe<SUB>2</SUB> (22) (prepared using 8), and [amido-phosphine-amine]MCl<SUB>3</SUB> (23, M = Hf; 24, M = Zr; prepared using 9) were confirmed by X-ray crystallography. Most of the prepared complexes exhibited negligible or low activity for ethylene/1-octene copolymerization. The [amido-phosphine sulfide]Hf(CH<SUB>2</SUB>Ph)<SUB>3</SUB> complex (13) exhibited relatively high copolymerization activity (19 x 10<SUP>6</SUP> g/mol-Hf h); however, this activity was unsatisfactory compared to that of the related [amido-phosphine]Hf(CH<SUB>2</SUB>Ph)<SUB>3</SUB> complexes (up to 48 x 10<SUP>6</SUP> g/mol-Hf h).