• Title/Summary/Keyword: Animal Bioscience

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Effects of Caffeine on Maturation-Promoting Factor (MPF) Activity in Bovine Oocytes and on the Development of Somatic Cell Nuclear Transfer Embryos in White-Hanwoo

  • Lee, Joon-Hee;Lee, Hee-Gyu;Baik, Sang-Ki;Jin, Sang-Jin;Moon, Song-Yi;Eun, Hye-Ju;Kim, Tae-Suk;Ko, Yeoung-Gyu;Kim, Sung-Woo;Park, Hae-Geum;Park, Soo-Bong
    • Reproductive and Developmental Biology
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    • v.36 no.4
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    • pp.261-267
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    • 2012
  • The technique of SCNT is now well established but still remains inefficient. The in vitro development of SCNT embryos is dependent upon numerous factors including the recipient cytoplast and karyoplast. Above all, the metaphase of the second meiotic division (MII) oocytes have typically become the recipient of choice. Generally high level of MPF present in MII oocytes induces the transferred nucleus to enter mitotic division precociously and causes NEBD and PCC, which may be the critical role for nuclear reprogramming. In the present study we investigated the in vitro development and pregnancy of White-Hanwoo SCNT embryos treated with caffeine (a protein kinase phosphatase inhibitor). As results, the treatment of 10 mM caffeine for 6 h significantly increased MPF activity in bovine oocytes but does not affect the developmental competence to the blastocyst stage in bovine SCNT embryos. However, a significant increase in the mean cell number of blastocysts and the frequency of pregnant on 150 days of White-Hanwoo SCNT embryos produced using caffeine treated cytoplasts was observed. These results indicated that the recipient cytoplast treated with caffeine for a short period prior to reconstruction of SCNT embryos is able to increase the frequency of pregnancy in cow.

Aberrant Distributions of ICM Cells in Bovine Blastocysts Produced by Somatic Cell Nuclear Transfer

  • D. B. Koo;Y. K. Kang;Park, Y. H.;Park, J. S.;Kim, H. N.;D. S. Son;Y. M. Han;Lee, K. K.
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.20-20
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    • 2001
  • It has been reported that cloning cattle is inefficient. One of the problems was placental abnormality, finally resulting in fetal mortality after transfer of nuclear transfer (NT) bovine embryos. This study was focused on the allocations of embryonic cells to the inner cell mass (ICM) or to the trophectoderm(TE) in NT bovine blastocysts. Somatic cells were derived from a Day 45 fetus of gestation, individually transferred into enucleated oocytes and developed to the blastocyst stage in vitro. Differential staining was used to assess the qualify of blastocysts derived from NT, IVF and in vivo. Development rate of NT embryos to blastocysts (25.0%, 41/164) was similar to that of IVF embryos (28.7%, 49/171). The total cell number of NT blastocysts (101.3$\pm$45.9) was not different compared with that of IVF embryos (107.9$\pm$34.2, P>0.05), but was lower than in vivo embryos (122.5$\pm$21.6, P<0.05). Ratio of ICM/total cells was higher in NT embryos (51.6$\pm$ 18.6%) than in IVF and in vivo embryos (42.3$\pm$ 15.3% and 34.9$\pm$8.9%, respectively) (P<0.05). Most IVF (56.8%, 25/44) and in vivo blastocysts(80.8%, 21/26) was distributed in the proportion of ICM/total cells ranging from 20 to 40% group. However, most NT blastocysts was biased in the 40-60%(34.1%, 15/44) and >60% (31.8%, 14/44) groups. Our findings suggest that placental abnormalities or early fetal losses in the present cloning system may be due to aberrant allocation of NT embryos to the ICM cells.

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Intravenous administration of L-tryptophan stimulates gastrointestinal hormones and melatonin secretions: study on beef cattle

  • Lee, Sang-Bum;Lee, Kyung-Won;Wang, Tao;Lee, Jae-Sung;Jung, U-Suk;Nejad, Jalil Ghassemi;Oh, Young-Kyoon;Baek, Youl-Chang;Kim, Kyoung Hoon;Lee, Hong-Gu
    • Journal of Animal Science and Technology
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    • v.61 no.4
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    • pp.239-244
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    • 2019
  • This study aimed to determine the effective dose of intravenous administration of L-tryptophan (L-T) on gastrointestinal hormones (GIH) secretions and melatonin using Hanwoo cattle. Three steers ($362{\pm}23kg$) fitted with indwelling jugular vein catheters were assigned in a $3{\times}3$ Latin square design. Treatments were intravenous administration of saline (control), 28.9 mg L-T/kg body weight (BW; low) and 57.8 mg L-T/kg BW (high) L-T for 1 day with 7 days of adaptation. Samples were collected after adaptation period at -60, 0, 30, 60, 90, 120, 150, 180, 240, and 300 min of sampling day. The levels of serum cholecystokinin (CCK) and secretin were higher (p < 0.05) in the high L-T group than those in the other groups. Serum Melatonin (MEL) levels were increased upon L-T administration (p < 0.05) in the high L-T group. Taken together, the effective dose of L-T administration was defined at 57.8 mg L-T/kg BW in order to stimulate increase of GIH and MEL.

Behavioral Characteristics of Weaned Piglets Mixed in Different Groups

  • Hwang, Hyun-Su;Lee, Jae-Kang;Eom, Tae-Kyung;Son, Seung-Hun;Hong, Joon-Ki;Kim, Ki-Hyun;Rhim, Shin-Jae
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.7
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    • pp.1060-1064
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    • 2016
  • With regard to animal welfare concerns, behavioral information of weaned and mixed piglets is great interest in swine production. The aim of this study was to demonstrate the change in behavior of weaned piglets over time in two different groups (littermates and piglets from different litters) after mixing. Two weaned groups of piglets (72 individuals in all) housed either with littermates or with foreign piglets (6 individuals in $1.8m{\times}1.4m$ pens, $28^{\circ}C{\pm}1^{\circ}C$ temperature) were observed with the aid of video technology for 9 consecutive hours on days 1, 2, and 3 after mixing. The behaviors of the weaned piglets in the control and treatment groups were significantly different among the days after mixing. Piglets were, however, more active and aggressive in the groups with foreign piglets. This study reveals a lower level of agonistic behavior in groups of piglets that came from the same litter.

Characterization of Mouse Peroxiredoxin III Genomic DNA and Its Expression

  • Lee, Tae-Hoon;Kim, Sun-Uk;Lee, Kyung-Kwang;Yu, Dae-Yeul
    • Proceedings of the KSAR Conference
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    • 2002.06a
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    • pp.64-64
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    • 2002
  • Peroxiredoxins (Prxs) possess protective activity against oxygen radicals generated by thiol-catalyzed oxidative systems. We already reported the genomic structure and its expression of mouse Prx Ⅰ, Ⅱ, and 1-Cys Prx. However, the Prx Ⅲ has not been determined. That was initially defined transiently expressed gene, mouse MER5, of murine erythroleukaemia cell differentiation. In addition, this protein was recently redefined a member of the thiol-specific antioxidant gene family. (omitted)

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Nutritional and Hormonal Induction of Fatty Liver Syndrome and Effects of Dietary Lipotropic Factors in Egg-type Male Chicks

  • Choi, Y.I.;Ahn, H.J.;Lee, B.K.;Oh, S.T.;An, B.K.;Kang, C.W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.8
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    • pp.1145-1152
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    • 2012
  • This experiment was conducted with male chicks to investigate the influence of hormones and nutrients on the development of fatty liver syndrome (FLS) as well as the effects of dietary lipotropic factors on hepatic fat accumulation and lipogenic enzyme gene expression. A total of two-hundred sixteen 4-wk-old Hy-Line male chicks were divided into six groups and fed an experimental diet (T1, low-energy diet with low levels of lipotropic factors; T2, high-energy diet with low levels of lipotropic factors; T3 and T5, low-energy diet with high levels of lipotropic factors; T4 and T6, high-energy diet with high levels of lipotropic factors) for six weeks. The chicks in T5 and T6 groups were treated with intramuscular injections of estradiol benzoate for three days prior to biopsy and clinical analysis of FLS. Chicks treated with estrogen had significantly greater liver weights than untreated chicks. The abdominal fat contents were increased in chicks consuming high-energy diets as compared to those consuming low-energy diets. Treatment with estrogen significantly increased the concentrations of serum cholesterol, triacylglycerol and phospholipid (p<0.05). The hepatic triacylglycerol levels were tenfold higher in the estrogen treated chicks than in the untreated chicks. There were no significant differences in malondialdehyde levels between the treatment groups. Estrogen treatment dramatically increased the levels of fatty acid synthetase, acetyl-CoA carboxylase and ApoB mRNA. The results indicated that treatment with exogenous estrogen in growing male chicks induced hepatic fat accumulation, which might be partially due to increased lipogenic enzyme gene expression.

Production of Cloned Bovine Embryos Carrying with Human Thrombopoietin Gene

  • K.I. Wee;B.H. Son;Park, Y.H.;Park, J.S.;D.H. Ko;Lee, K.K.;Y.M. Han
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.60-60
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    • 2001
  • Human thrombopoietin (hTPO) is a cytokine that plays a central role in megakaryopoiesis by influencing on the development and maturation of megakaryocyte and platelet production. To induce hTPO production in the mammary gland, expression vector was constructed by combining the promoter of bovine beta-casein gene, cDNA of hTPO and neomycine resistance gene for transfection into fibroblasts. Bovine fibroblast cells derived from female ear skin were transfercted with the expression vector using Lipofectamine (Life Technology, NY). Transected cells resistant to G4l8 treatment (600 $\mu\textrm{g}$/$m\ell$) were recovered and colony formation was initiated at 13 days. The colonies with about 1 cm diameter were picked and analysed by PCR. Single transfected cells were individually transferred to enucleated oocytes. After electrofusion, the reconstructed embryos were exposed to calcium ionophore (5uM) for 5 min followed by treatment with 6-DMAP (2.5 mM) for 4h. The nuclear transfer embryos were cultured in CRlaa medium at 38.5C, 5% $CO_2$ for 7 days. Twenty three of 29 (79.3%) colonies were proved to be hTPO transfectants by PCR. The colonies were further passaged and used to produce transgenic embryos using nuclear transfer. Cleavage and developmental rates of reconstructed embryos to the blastocyst stage were 65.1% and 39.4%, respectively Of 22 blastocysts that developed from reconstructed embryos with the transfected cell, 20 embryos (90.9%) were positive for hTPO by using PCR analysis. The results suggest that somatic cell nuclear transfer is efficient for production of transgenic embryos.

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Limited in vitro differentiation of porcine induced pluripotent stem cells into endothelial cells

  • In-Won Lee;Hyeon-Geun Lee;Dae-Ky Moon;Yeon-Ji Lee;Bo-Gyeong Seo;Sang-Ki Baek;Tae-Suk Kim;Cheol Hwangbo;Joon-Hee Lee
    • Journal of Animal Reproduction and Biotechnology
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    • v.38 no.3
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    • pp.109-120
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    • 2023
  • Background: Pluripotent stem cells (PSCs) including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) offer the immense therapeutic potential in stem cell-based therapy of degenerative disorders. However, clinical trials of human ESCs cause heavy ethical concerns. With the derivation of iPSCs established by reprogramming from adult somatic cells through the transgenic expression of transcription factors, this problems would be able to overcome. In the present study, we tried to differentiate porcine iPSCs (piPSCs) into endothelial cells (ECs) for stem cell-based therapy of vascular diseases. Methods: piPSCs (OSKMNL) were induced to differentiation into ECs in four differentiation media (APEL-2, APEL-2 + 50 ng/mL of VEGF, EBM-2, EBM-2 + 50 ng/mL of VEGF) on cultured plates coated with matrigel® (1:40 dilution with DMEM/F-12 medium) for 8 days. Differentiation efficiency of these cells were exanimated using qRT-PCR, Immunocytochemistry, Western blotting and FACS. Results: As results, expressions of pluripotency-associated markers (OCT-3/4, SOX2 and NANOG) were higher observed in all porcine differentiated cells derived from piPSCs (OSKMNL) cultured in four differentiation media than piPSCs as the control, whereas endothelial-associated marker (CD-31) in the differentiated cells was not expressed. Conclusions: It can be seen that piPSCs (OSKMNL) were not suitable to differentiate into ECs in the four differentiation media unlike porcine epiblast stem cells (pEpiSCs). Therefore, it would be required to establish a suitable PSCs for differentiating into ECs for the treatment of cardiovascular diseases.

Immune Modulation Effect of Pig Placenta Extracts in a Mouse Model: Putative Use as a Functional Food Supplement

  • Park, Hyun-Jung;Suh, Han-Geuk;Kim, Jin-Hoi;Jang, Ae-Ra;Jung, Hyun-Jung;Lee, Sung-Dae;Ha, Woo-Tae;Lee, Ran;Kim, Ji-Hyuk;Kim, Sang-Ho;Sung, Si-Heung;Moon, Sang-Ho;Kim, Bo-Kyung;Song, Hyuk
    • Food Science of Animal Resources
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    • v.31 no.5
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    • pp.701-709
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    • 2011
  • This study was performed to establish an effective extraction method of pig placenta extract that could be used for a putative functional food supplement with immunomodulatory effects. In the present study, we used different temperatures (4, 37, 60, 80, and $100^{\circ}C$) and different solvents (chloroform, NaOH, and phosphate buffered saline [PBS]) to extract the pig placenta. Among the different placenta extracts yielded by the different extraction methods, placenta extract (PE) in PBS at $80^{\circ}C$ for 30 min (referred to as PE-PBS80) showed a significant increase of nitric oxide production of up to 22.97 ${\mu}M/10^5$ cells at a 1 mg/mL dose (p<0.05 ) in J774A.1 cells than other extracts and control tested. Using PE-PBS80, further animal challenges were performed to identify the immune-enhanced effects. As a result, orally administered PE-PBS80 showed a significant increase in blood T and B cell activities and immunoglobulin (IgG and IgM) production. IgG and IgM levels increased to 41.53 mg/mL at a 20 mg dose on day 7 and to 27.38 mg/mL at a 10 mg dose on day 14, respectively (p<0.05). Furthermore, PE-PBS80 was also able to significantly enhance the immune modulator cytokine levels (p<0.05) compared to the control and vehicle treatments. Among the evaluated cytokines, the tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) level increased to 28.89 pg/mL at extract doses of 20 and 50 mg, the interleukin-$1{\beta}$ (IL-$1{\beta}$) level increased to 21.52 pg/mL at extract doses of 10, 20, 50 and 75 mg and the interferon (IFN)-${\gamma}$ level increased to 18.24 pg/mL at extract doses of 10, 20, and 50 mg. Therefore, this study presents an effective method for extracting pig placenta extracts and also demonstrates that pig placenta extracts had significant immunomodulatory effects not only at the cellular level but also in a mouse model, suggesting that this material could be used as an excellent candidate functional food supplement.

Development of a New Type of Recombinant Hyaluronidase Using a Hexahistidine; Possibilities and Challenges in Commercialization

  • Park, Chaeri;Song, Yun-Kyoung;Kim, Young-Hyun;Jung, Yena;Park, Young-Ho;Song, Bong-Seok;Eom, Taekil;Kim, Ju-Sung;Kim, Sang-Hyun;Kim, Ji-Su;Kim, Sun-Uk;Lee, Sang-Rae;Kim, Ekyune
    • Journal of Microbiology and Biotechnology
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    • v.29 no.8
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    • pp.1310-1315
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    • 2019
  • Hyaluronidases enhance therapeutic drug transport by breaking down the hyaluronan barrier to lymphatic and capillary vessels, facilitating their tissue absorption. Commercially available hyaluronidases are bovine in origin; however, they pose risks such as bovine spongiform encephalopathy. The present study aimed to develop a novel, highly active hyaluronidase and assess its function. Therefore, in order to find the most efficient active hyaluronidase, we produced several shortened hyaluronidases with partial removal of the N- or C-terminal regions. Moreover, we created an enzyme that connected six histidines onto the end of the hyaluronidase C-terminus. This simplified subsequent purification using $Ni^{2+}$ affinity chromatography, making it feasible to industrialize this highly active recombinant hyaluronidase which exhibited catalytic activity equal to that of the commercial enzyme. Therefore, this simple and effective isolation method could increase the availability of recombinant hyaluronidase for research and clinical purposes.