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miR-375 down-regulation of the rearranged L-myc fusion and hypoxia-induced gene domain protein 1A genes and effects on Sertoli cell proliferation

  • Guo, Jia;Liu, Xin;Yang, Yuwei;Liang, Mengdi;Bai, Chunyan;Zhao, Zhihui;Sun, Boxing
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.8
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    • pp.1103-1109
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    • 2018
  • Objective: This study aimed to screen and identify the target genes of miR-375 in pig Sertoli (ST) cells and to elucidate the effect of miR-375 on the proliferation of ST cells. Methods: In this study, bioinformatics software was used to predict and verify miR-375 target genes. Quantitative polymerase chain reaction (PCR) was used to detect the relationship between miR-375 and its target genes in ST cells. Enzyme-linked immunosorbent assay (ELISA) of rearranged L-myc fusion (RLF) and hypoxia-induced gene domain protein 1A (HIGD1A) was performed on porcine ST cells, which were transfected with a miR-375 mimics and inhibitor to verify the results. Dual luciferase reporter gene assays were performed to assess the interactions among miR-375, RLF, and HIGD1A. The effect of miR-375 on the proliferation of ST cells was analyzed by CellTiter 96 AQueous One Solution Cell Proliferation Assay (MTS). Results: Five possible target genes of miR-375, including RLF, HIGD1A, colorectal cancer associated 2, POU class 3 homeobox 1, and WW domain binding protein 1 like, were found. The results of quantitative PCR suggested that mRNA expression of RLF and HIGD1A had a negative correlation with miR-375, indicating that RLF and HIGD1A are likely the target genes of miR-375. The ELISA results revealed that RLF and HIGD1A were negatively correlated with the miR-375 protein level. The luminescence results for the miR-375 group cotransfected with wild-type RLF and HIGD1A vector were significantly lower than those of the miR-375 group co-transfected with the blank vector or mutant RLF and HIGD1A vectors. The present findings suggest that RLF and HIGD1A are target genes of miR-375 and that miR-375 inhibits ST cell proliferation according to MTS analysis. Conclusion: It was speculated that miR-375 affects cell proliferation through its target genes, which play an important role in the development of testicular tissue.

Effects of dietary supplementation of fermented wheat bran on performance and blood profiles in weaned pigs

  • Jeong, Yong Dae;Lee, Jung Jae;Kim, Jo Eun;Kim, Doo Wan;Min, Ye Jin;Cho, Eun Seok;Yu, Dong Jo;Kim, Young Hwa
    • Korean Journal of Agricultural Science
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    • v.44 no.3
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    • pp.409-415
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    • 2017
  • This study was conducted to investigate the effect of supplementation of fermented wheat bran (FWB) on growth and blood characteristics in weaned pig. A total of 36 weaned pigs ($Landrace{\times}Yorkshire{\times}Duroc$; BW, $7.78{\pm}0.04kg$) were randomly allocated to three dietary treatments with different FWB concentrations (0, 0.5, and 1.0%), and each treatment had 3 replicate pens with 4 pigs per pen. The FWB was obtained from a mixture of wheat bran and two microbes (Lactobacillus plantarum M10 and Saccaromyces cerevisiae) and was determined to contain $10.19{\pm}0.27log\;CFU/g$ of L. plantarum and $7.73{\pm}0.38log\;CFU/g$ of S. cerevisiae. Experimental diets were prepared by mixing 0 (control), 0.5, or 1.0% of the FWB to the basal diet, and fed to the weaned pigs for 7 weeks. During the experimental period, the pigs had access to the diet and water ad libitum. Feed intake increased significantly in the 1.0% FWB group compared to the control and 0.5% FWB groups (p < 0.05), whereas the other growth parameters were not different among the treatment groups. White blood cells and lymphocytes were significantly decreased in the FWB treatment groups compared to the control group, but other blood corpuscles were not different among the treatment groups (p < 0.05). The pigs fed 0.5% FWB showed greater serum IgG than the control and 1.0% FWB groups (p < 0.05). In conclusion, the FWB fed to weaned pigs did not negatively affect their growth performance, but rather reduced mortality by fortifying immunity.

Pharmacological Actions of New Woohwangchungsimwon Pill on Cardiovascular System (신우황청심원의 심혈관계에 대한 약효)

  • Cho, Tai-Soon;Lee, Sun-Mee;Kim, Nak-Doo;Huh, In-Hoi;Ann, Hyung-Soo;Kwon, Kwang-Il;Park, Seok-Ki;Shim, Sang-Ho;Shin, Dae-Hee;Park, Dai-Kyu
    • YAKHAK HOEJI
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    • v.41 no.6
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    • pp.802-816
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    • 1997
  • In order to investigate the pharmacological properties of New Woohwangehungsimwon Pill (NWCH). Effects of Woohwangehungsimwon Pill (WCH) and NWCH were compared using various experimental models. In isolated rat aorta, NWCH and WCH showed the relaxation of blood vessels in maximum contractile response to phenylephrine ($10^{-6}$M) without regard to endothelium containing or denuded rings of the rat aorta. Furthermore, the presence of the inhibitors of NO synthase and guanylate cyclase did not affect significantly the relaxative effects of NWCH and WCH. NWCH and WCH inhibited the vascular contractions induced by acethylcholine, prostaglandin endoperoxide or peroxide in a dose-dependent manner. In conscious spontaneously hypertensive rats(SHRs), NWCH and WCH decreased significantly heart rate. These, at high doses, had a negative inotropic effect that was a decrease of LVDP and (-dp/dt)/(+dp/dt) in the isolated perfused rat hearts, and also decreased the contractile force and heart rate in the isolated rat right atria. In excised guinea-pig papillary muscle, these had no effects on parameters of action potential at low doses, whereas inhibited the cardiac, contractility at high doses. Furthermore, these had a significant inhibitory effects on heart acceleration in normotensive rats and SHRs. These results suggested that NWCH and WCH have weak cardiovascular effects, and that there is no significant differences between two preparations.

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Immunological Characterization of Full and Truncated Recombinant Clones of ompH(D:4) Obtained from Pasteurella multocida (D:4) in Korea

  • Kim, Young-Hwan;Cheong, Ki-Young;Shin, Woo-Seok;Hong, Sung-Youl;Woo, Hee-Jong;Kwon, Moo-Sik
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1529-1536
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    • 2006
  • We cloned a gene of ompH(D:4) from pigs infected with P. multocida D:4 in Korea [16]. The gene is composed of 1,026 nucleotides coding 342 amino acids (aa) with a signal peptide of 20 aa (GenBank accession number AY603962). In this study, we analyzed the ability of the ompH(D:4) to induce protective immunity against a wild-type challenge in mice. To determine appropriate epitope(s) of the gene, one full and three different types of truncated genes of the ompH(D:4) were constructed by PCR using pET32a or pRSET B as vectors. They were named ompH(D:4)-F (1,026 bp [1-1026] encoding 342 aa), ompH(D:4)-t1 (693 bp [55-747] encoding 231 aa), ompH(D:4)-t2 (561 bp [187-747] encoding 187 aa), and ompH(D:4)-t3 (540 bp [487-1026] encoding 180 aa), respectively. The genes were successfully expressed in Escherichia coli BL21(DE3). Their gene products, polypeptides, OmpH(D:4)-F, -t1, -t2, and -t3, were purified individually using nickel-nitrilotriacetic acid (Ni-NTA) affinity column chromatography. Their $M_rs$ were determined to be 54.6, 29, 24, and 23.2 kDa, respectively, using SDS-PAGE. Antisera against the four kinds of polypeptides were generated in mice for protective immunity analyses. Some $50{\mu}g$ of the four kinds of polypeptides were individually provided intraperitoneally with mice (n=20) as immunogens. The titer of post-immunized antiserum revealed that it grew remarkably compared with pre-antiserum. The lethal dose of the wild-type pathogen was determined at $10{\mu}l$ of live P. multocida D:4 through direct intraperitoneal (IP) injection, into post-immune mice (n=5, three times). Some thirty days later, the lethal dose ($10{\mu}l$) of live pathogen was challenged into the immunized mouse groups [OmpH(D:4)-F, -t1, -t2, and -t3; n=20 each, two times] as well as positive and negative control groups. As compared within samples, the OmpH(D:4)-F-immunized groups showed lower immune ability than the OmpH(D:4)-t1, -t2, and -t3. The results show that the truncated-OmpH(D:4)-t1, -t2, and -t3 can be used for an effective vaccine candidate against swine atrophic rhinitis caused by pathogenic P. multocida (D:4) isolated in Korea.

Associations of the Porcine Melanocortin-4 Receptor (MC4R) Gene with Growth Traits in Duroc Pigs (듀록 품종의 Melanocortin-4 Receptor(MC4R) 유전자와 성장형질과의 연관성 분석)

  • Cho, K.H.;Kim, M.J.;Choi, B.H.;Jeon, G.J.;Ryu, J.W.;Jung, H.J.;Kim, I.C.;Lee, H.K.;Jeon, G.J.
    • Journal of Animal Science and Technology
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    • v.49 no.4
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    • pp.437-442
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    • 2007
  • The melanocortin-4 receptor(MC4R) is virtually expressed in all brain regions and plays an important role in energy homeostasis in mammals. MC4R has been intensively studied as a trait gene controlling economically important traits, such as growth and feed conversion, etc. Six hundreds and sixty Duroc pigs were genotyped for the MC4R locus and analyzed their relationships with breeding values for average daily gain(ADG), backfat thickness(BF), days to 90kg(D90) and feed conversion(FC). The estimated genotype frequencies for the all Duroc pigs were: 30.8%, 45.2%, 24.1% for AA, AB and BB genotypes, respectively. The mutant A allele was significantly associated with ADG, D90 and BF whereas no significant relationship was found with FC. The change of gene frequencies by generation was shown in both selected and culled groups. These results indicate that the MC4R polymorphism could be integrated in the present selection program to realize a marker-assisted selection for the growth traits of the Duroc population.

Effects of Dietary Supplementation of Fermented Microbial Complex (Eco-Farm$^{(R)/}$) on Performance of Finishing Pigs and Air Quality in Finishing Building (복합미생물 발효제(Eco-Farm$^{(R)}$ ) 급여가 비육돈 생산성과 돈사 환경개선에 미치는 영향)

  • 김두환;정치섭
    • Journal of Animal Environmental Science
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    • v.9 no.1
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    • pp.27-34
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    • 2003
  • This experiment was carried out to investigate the effect of dietary supplementation of fermented microbial complex(Eco-Farm$^{(R)}$) on performance of finishing pigs and indoor air quality in finishing building. A total 135 crossbred [(Landrace ${\times}$ Yorkshire) ${\times}$ Duroc] pigs were randomly arranged into nine groups and assigned to three treatments. Pigs were fed a basal diet supplemented with 0, 0.5 and 1% level of fermented microbial complex(Eco-Farm$^{(R)}$) until the market weight for 40 days of the experimental period. Average daily feed intake and feed conversion ratio were significantly improved (p<0.05) with dietary supplementation of 0.5% fermented microbial complex (Eco-Farm$^{(R)}$): however, average daily gain was not affected by dietary supplementation of fermented microbial complex(Eco-Farm$^{(R)}$). Indoor ammonia and hydrogen sulfide concentrations in the finishing building were significantly(p<0.05) decreased by dietary supplementation of fermented microbial complex(Eco-Farm$^{(R)}$) compared with those of control, however, indoor carbon dioxide concentration was not affected by dietary supplementation of fermented microbial complex(Eco-Farm$^{(R)}$). In conclusion, the results obtained from this experiment suggest that the dietary supplementation of fermented microbial complex(Eco-Farm$^{(R)}$) for finishing pigs improved performance and indoor air quality in the finishing building.hing building.

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Study on the Lowering of NaCl Content by Co-composting Food Wastes (저농도 식염을 함유하는 음식물쓰레기 퇴비제조)

  • Lee, In-Bog;Park, Chang-Kyu;Kim, Pil-Joo
    • Korean Journal of Soil Science and Fertilizer
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    • v.34 no.1
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    • pp.17-25
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    • 2001
  • High salt content in food waste limit use as the potential source of compost. Since sodium chloride content should be lower than one percent to meet requirement for agricultural use of compost, we attempted use of three kinds of co-composting material for food wastes compost, which were fermented, dried, and fresh types of amendments which included pig manure, sawdust and puffed rice hull. Food wastes were composted well and stabilized after around the 40th days of composting. Little difference were found between composts amended sawdust and puffed rice hull. The result indicated that puffed rice hull could be used as a good alternative of sawdust. Because sawdust or puffed rice hull were mixed as amendments to control water contents and to dilute high NaCl content of compost material, the C/N ratios of the final products were significantly high over 40. However, NaCl contents of final products were dropped to less than percent on the fresh weight basis by mixing food wastes and the above amendments with the optimum rates.

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Effects of coated cysteamine hydrochloride on muscle fiber characteristics and amino acid composition of finishing pigs

  • Bai, Miaomiao;Liu, Hongnan;Xu, Kang;Yu, Rong;Oso, Abimbola Oladele;Deng, Jinping;Yin, Yulong
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.9
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    • pp.1430-1438
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    • 2019
  • Objective: This experiment was designed to determine the effects of coated cysteamine hydrochloride (CC) on muscle fiber characteristics, amino acid composition and transporters gene expression in the longissimus dorsi muscle (LDM) of finishing pigs. Methods: Two hundred and sixteen Duroc/Landrace/Yorkshire cross-bred male finishing pigs were fed with a corn-soybean basal diet supplemented with 0, 70, and 140 mg/kg cysteamine. Each group contained eight replicates of nine pigs per replicate. After 29 days, one pig was randomly selected from each replicate and slaughtered. Blood and LDM samples were collected and analyzed. Results: The results showed that supplemental dietary CC increased (p<0.05) the muscle fiber density. And CC supplementation also up-regulated (p<0.05) the expression of myosin heavy chain 1 (MyHC1) and MyHC2x mRNA levels, and down-regulated (p<0.05) MyHC2b expression in the LDM. Additionally, supplemental dietary CC reduced (p<0.05) the concentration of total cholesterol in the plasma and enhanced (p<0.05) the concentrations of essential amino acid and total amino acid in the LDM. The relative expression levels of chloramphenicol acetyltransferase 2, $b^{0,+}$ amino acid transporter, and $y^+$-L-type amino acid transporter 1 were upregulated (p<0.05) in the LDM when pigs were fed with the dietary CC of 70 mg/kg. Conclusion: Cysteamine supplementation could increase fiber density and distribution of fiber types. It also improved the deposition of protein in the LDM by up-regulated the expression of amino acid transporters.

Clinical characterization of 3-month-old pigs infected with African swine fever virus from Vietnam

  • Oh, Sang-Ik;Bui, Vuong Nghia;Dao, Duy Tung;Bui, Ngoc Anh;Yi, Seung-Won;Kim, Eunju;Lee, Han Gyu;Bok, Eun-Yeong;Wimalasena, S.H.M.P;Jung, Young-Hun;Hur, Tai-Young;Lee, Hu Suk
    • Korean Journal of Veterinary Service
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    • v.45 no.2
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    • pp.71-77
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    • 2022
  • African swine fever (ASF) is a fatal viral disease in pigs, with a short incubation period and causing immediate death. Few studies exist on the Asian epidemic ASF virus (ASFV) challenge in older pigs, including growing and fattening pigs and sows. We aimed to investigate clinical outcomes, pathomorphological lesions, and viral distribution in organs of 3-month-old growing pigs that were inoculated with the ASFV isolated in Vietnam. The clinical outcomes were recorded daily, and the dead or euthanized pigs immediately underwent necropsy. Viral loads were determined in 10 major organs using quantitative polymerase chain reaction. The average incubation period in growing pigs was more delayed (5.2±0.9 dpi) than that in weaned pigs, and the clinical signs were milder in growing pigs than in weaned pigs. The digestive and respiratory clinical signs in growing pigs showed at the end period of life, but these were observed at an early stage of infection in weaned pigs. The pathomorphological features were severe and nonspecific with hemorrhagic lesions in various organs. The viral loads in organs from growing pigs were higher than those from piglets, and the number of viral copies was related to gross lesions in the tonsil and intestine. In the absence of vaccines against ASF, early clinical detection is important for preventing the spread of the virus. Our findings elucidated that the clinical signs and gross lesions in growing pigs differed from those in weaned pigs, which provide valuable information for diagnosis of pigs with suspected ASF infection.

Morphological Characteristics of Pig Blastocysts Produced by Somatic Cell Nuclear Transfer

  • Y.M. Han;D.B. Koo;Park, Y.H.;Park, J.S.;Kim, H.N.;Y.K. Kang;W.K. Chang;Lee, K.K.
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.68-68
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    • 2001
  • Blastocyst formation, consisting of the inner cell mass (ICM) and trophectoderm (TE), is the first differentiation process during embryonic development in mammals. It has been hypothesized that the proportion of ICM to TE in the blastocyst may be crucial for subsequent developmental competence of early embryos, which it may be expressed as a sensitive indicator for evaluating in vitro systems. In this study ICM/total cell ratio of nuclear transfer (NT) embryos was compared with IVF-derived and in vivo embryos. Somatic cell nuclei obtained from a fetus at Day 40 of gestation were transferred into the enucleated oocyte and then cultured in NCSU 23 medium for 6 days as previously described (Koo et al., Biol. Reprod. 2000; 63:986-992). ICM and TE cells of blastocysts were determined by using a differential staining method (Han et al., Biol. Reprod. 1999; 60:1110-1113). Development rate (9.8$\pm$2.5%, 23/225) to the blastocyst stage of NT embryos was lower than IVF embryos (23.8$\pm$2.7%, 53/223). Thus, a difference was detected in the in vitro developmental rate to blastocyst stage between NT and IVF-derived embryos (P<0.05). In the next experiment, we investigated ICM and TE nuclei to assess the quality of blastocysts that produced by NT, IVF and in vivo, respectively. NT blastocysts (27.6$\pm$8.3) showed a smaller total cell number than IVF-derived (42.6$\pm$17.4) and in vivo embryos (283.9$\pm$103.5) (P<0.05). Ratios of ICM/total cells in NT, IVF and in vivo blastocysts were 15.1$\pm$ 18.6% (n=56), 12.3$\pm$9.2% (n=57) and 30.4$\pm$6.8% (n=40), respectively. Individual blastocysts for the ratio of ICM/total cells were assigned to 3 groups (I; <20%, II; 20 to 40% and III;>40%). As the results, most in vivo blastocysts (97.5%, 39/40) were distributed into group II while most NT (78.6%, 44/56) and IVF-derived blastocysts (82.5%, 47/57) were allocated to group I. Thus, our data show that NT or IVF-derived embryos have aberrant morphology during early development in vitro systems, suggesting that these anomalies may result in developmental failures of the NT embryos to term.

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