• Title/Summary/Keyword: C6 세포

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Studies on In vitro Fertilization and Development of Bovine Follicular Oocytes Matured In vitro I. Effect of Cumulus Cells on In vitro Fertilization and Development of Bovine Follicular Oocytes Matured In vitro (체외성숙 우난포란의 체외수정과 발달에 관한 연구 I. 난구세포가 체외성숙 우난포란의 체외수정과 발달에 미치는 영향)

  • Park, S.P.;Kim, E.Y.;Chung, H.M.;Chung, K.S.
    • Korean Journal of Animal Reproduction
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    • v.14 no.1
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    • pp.1-8
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    • 1990
  • These experiments were carried out to investigate the effects of cumulus cells for in vitro fertilization and development of bovine follicular oocytes matured in vitro. The bovine ovaries were obtained at a slaughter house and the follicular oocytes surrounded by cumulus cells were collected by puncturing follicles with 2~6 mm of diameter. Bovine oocytes were matured in vitro for 24~26 hours in a CO2 incubator with 5% CO2 in air at 39$^{\circ}C$. The medium used for maturation was TCM-199 supplemented with hormones, pyruvate, FCS and antibiotics. Epididymal spermatozoa were capacitated by in vitro culture for 2~3 hours in BO solution containing BSA(5mg/ml) and caffeine(2.5mM). Insemination was made by introducing about 10~15 matured oocytes into the suspension of capacitated spermatozoa. Six hour after insemination the eggs were transferred to TCM-199 supplemented with FCS(10%) and HEPES(25mM), cultured for 7~8 days with 10~15 eggs/well in 4-well multidishes(Nunc Co.) forming cumulus cell monolayer. The results obtained in these experiments were summarized as follows ; 1. The majority of the follicular oocytes with compacted cumulus cells existed in GV stage while those with dispersed or denuded cumulus cells existed GVBD and M II stage. 2. After 24~26 hours maturation, the maturation rates of the follicular oocytes cultured in TCM-199 containing hormones were slightly higher than those of oocytes cultured in medium without hormones, and the frequency of cumulus compacted or denuded oocytes reaching M II stage cultured in medium containing hormones was 75.7% or 51.7%, respectively(P<0.05). 3. After 20 hours in vitro insemination, percentages of ova fertilized were 61.4% or 51.4%, respectively, for cumulus oophorus intacted or removed, and increased frequency of ova with both male and female pronuclei was found when cumuli were present(P<0.05). 4. The rates of embryos developed to 2-, 4-, 8-, 16-cell and morula or blastocyst stage after cocultured with cumulus cells were 65.0%, 45.3%, 34.7%, 28.0% and 22.7%, respectively. The results for momla or blastocyst stage were significantly higher than those of the embryos cultured in the basic medium(P<0.05).

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Patterns of Middle School Students' Value-Judgement and Decision-Making on Biotechnology-Related Socioscientific Issues (생명공학과 관련된 사회적 쟁점에 대한 중학생들의 가치판단 및 의사결정 과정에서 드러난 가치 적용 방식 유형)

  • Ju, In-Ae;Lee, Hyunju
    • Journal of The Korean Association For Science Education
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    • v.33 no.1
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    • pp.79-93
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    • 2013
  • This study examined value-judgement and decision-making process of middle school students on three biotechnology related socioscientific issues (SSI) (i.e. designer baby, amniotic fluid test, and embryonic stem cell research). In particular, it focused on the level of consistency in their value application to the decisions. Twenty 8th grade students participated in the SSI program. The participants were asked to mark their positions on each issue before and after the debates. All classroom and sub-group debates were audio-recorded. The researcher conducted individual interviews with 10 students after the program. The results show three patterns emerging. First, five students applied their personal values in a consistent manner when making decisions on the three consecutive issues. Second, three students demonstrated incremental resoluteness in their value application as the sessions proceeded and had tendency to be oriented towards a specific value, more so towards the end of the program than the beginning. And third, four students gradually showed ambivalence in their decisions as they participate in the program. The results suggest that science teachers addressing SSI need to encourage students to be aware of their own values and to engage in internalized decision-making in light of those values. They also need to be aware of the possible challenges that students might feel in their decision-making process caused by the complexity of SSI.

Molecular Cloning and mRNA Expression of the Bovine Peroxisome Proliperator Receptor Gamma(PPARγ) (한우 PPARγ 유전자의 동정과 mRNA의 발현)

  • Jeoung, Y.H.;Lee, S.M.;Park, H.Y.;Yoon, D.H.;Choi, J.G.;Moon, S.J.;Kang, M.J.
    • Journal of Animal Science and Technology
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    • v.46 no.1
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    • pp.23-30
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    • 2004
  • The peroxisome proliferator-activated receptor $\gamma$(PPAR$\gamma$), a member of the steroid/thyroid nuclear hormone receptor suferfamily of ligand-activated transcription factor, is an important regulator of adipocyte gene expression and differentiation. In this studies, we report the identification, characterization, and expression of a Hanwoo PPAR$\gamma$ gene. The PPAR$\gamma$ cDNA sequence of the Hanwoo show strong conservation with the corresponding sequences reported in other species except of three amino acid sequences. The distribution of PPAR$\gamma$ mRNA in various tissues of Korean cattle aged 12 months were investigated using Northern Blot analysis. The highest expression was detected in adipose tissue, more lower expression was detected in colon, small intestine, kidney, lung, while expression was not detected in brain, heart. PPAR$\gamma$ expression was higher in adipose tissue of Korean cattle when aged 30 months than aged 12 months. These results indicated PPAR$\gamma$, regulator adipocyte gene expression and differentiation, related on adipose differentiation in Korean native cattle(HANWOO).

Anti-metastatic Activity of Water Extract of Samguikoeuitang via Enhancement of Natural Killer Cells (삼귀고의탕 물추출물의 자연살해세포 활성을 통한 항전이효과)

  • Ko, Hyun-Suk;Park, Moon-Nyeo;Kim, Bong-Lee;Yoon, Taek-Joon;Song, Hyo-Sook;Cho, Sung-Yun;Lee, Hyo-Jung;Lee, Hyo-Jeong;Jeong, Soo-Jin;Lee, Eun-Ok;Kim, Sung-Hoon
    • Journal of Korean Traditional Oncology
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    • v.15 no.1
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    • pp.29-36
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    • 2010
  • The anti-metastatic activity of water extract of Samguikoeuitang (WSGKE) consisting of Ginseng Radix, Angelicae Radix, Sophora flavescens and Coicis Semenwas examined. Ethanol extract of Samguikoeuitang (ESGKE) showed significant cytotoxicity against colon 26-M3.1 carcinoma cells, while WSGEK did not. However, WSGKE significantly increased the production of IL-6 and IL-12 in thioglycollate-induced macrophages from Balb/c mice, whereas ESGKE did not. WSGKE significantly increased natural killer (NK) cell cytotoxicity against effecter YAC-1 cells in an Effecter cells/Target ratio dependent manner. Also, WSGKE significantly suppressed lung metastasis after i.v. injection of colon26-M3.1 carcinoma cells. Inhibitory effect of WSGKE on lung metastasis totally abolished in NK cells-deficient mice by treatment with anti-asialo GM1 serum. In addition, the combination treatment of cisplatin and WSGKE (100 ${\mu}g$/mouse) prolonged the lifespan of mice inoculated by colon26-M3.1 cell. These findings suggest that WSGKE can inhibit lung metastasis and prolong life span via immunological enhancement as a Biological Response Modifier.

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Characteristics of virus-like particles in color mutants of oyster mushrooms (원형느타리백색변이체에 존재하는 바이러스의 특성)

  • Lee, Kang-Hyo;Kim, Gyu-Hyun;Lee, Jin-Kyung;Seok, Soon-Ja;Weon, Hang-Yeon;Kim, Yang-Sup;Kim, Wan-Gyu;Sung, Jae-Mo
    • Journal of Mushroom
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    • v.5 no.1
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    • pp.39-42
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    • 2007
  • This study was carried out to characterize virus-like particles (VLPs) in Pleurotus ostreatus. The white and the dark gray mutants frequently observed in mushroom farms of Pleurotus ostreatus (Wonhyeong-neutari). A 5.8kb segments of dsRNA was detected only in the white mutants but not in the dark gray mutants. The VLPs were purified from the fruit bodies by Polyethylene Glycol (PEG) and ultracentrifugation. Electron microscopy analysis showed that VLPs were isometric about 14, 20~45nm in diameter. Further study is needed to reveal the morphological and yield variations of mushroom strains including VLPs observed in the mushroom farms. Also it is needed to maintain fundamental research for taxonomy, diagnosis, and physiology of VLPs in the mushroom strains.

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The spinal neuronal activity induced by low power laser stimulation (저출력 레이저 자극에 의한 척수내 신경세포의 활성변화)

  • Oh, Kyung-Hwan;Choi, Young-Deog;Lim, Jong-Soo
    • Journal of Korean Physical Therapy Science
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    • v.8 no.2
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    • pp.1005-1013
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    • 2001
  • The present study was designed to investigate the effect of low power GaAlAs laser on spinal Fos expression related to the anti-nociceptive effect of laser stimulation. Low power GaAlAs laser was applied to either acupoint or non-acupoint for 2 hour under light inhalation anesthesia. Spinal Fos expression in the dorsal horn was compared to that obtained in inhalation anesthesia control group. Furthermore, we analyzed the effect of the local treatment of lidocaine on the spinal Fos expression evoked by low power GaAlAs laser stimulation. The results were summarized as follows: 1. In the normal animals, only a few Fos like immunoreactive(Fos-IR) neurons were evident in the lumbar spinal cord dorsal horn. Similarly, following prolonged inhalation anesthesia, Fos-IR neurons were absent in the dorsal horn of the lumbar spinal cord. In animals treated with laser stimulation, Fos immunoreactive neurons were increased mainly in the medial half of ipsilateral laminae I-III at lumbar segments L3-5. These findings directly indicated that prolonged anesthesia used in this study did not affect the Fos expression in the spinal cord dorsal horn of intact animals and low power laser stimulation dramatically produced Fos expression in the spinal cord laminae that are related to the anti-nociceptive effect of laser stimulation. 2. In acupoint stimulated animals, 10mW of laser stimulation, not 3mW and 6mW intensity, significantly increased the number of Fos immunoreactive neurons in the spinal dorsal horn(p<0.05). However, laser stimulation on acupoint more dramatically increased the number of Fos immunoreactive neurons in the spinal cord rather than laser stimulatin on non acupoint. These result suggested that laser stimulatin on acupoint was more effective treatment to activate the spinal neuron than non acupoint stimulation. 3. The local treatment of lidocaine totally suppressed the activity of spinal neurons that were induced by lower power 1aser stimulation. These data indicated that the anti-nociceptive effect of laser stimulation was absolutely dependent upon the peripheral nerve activity in the stimulated location. In conclusion, these data indicate that 10mW of low power laser stimulation into acupoint is capable of inducing the spinal Fos expression in the dorsal horn related to the anti-nociceptive effect of laser stimulation, Furthermore, the induction of spinal Fos expression was totally related to the peripheral nerve activity in the laser stimulated area.

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The effect of phosphorus stress on the energy status and bacteroid content in soybean nodules (인산결핍이 대두근류의 bacteroid 함량과 energy 상태에 미치는 영향)

  • Sa, Tong-Min;Lim, Sun-Uk;Israel, Daniel W.
    • Applied Biological Chemistry
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    • v.35 no.6
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    • pp.449-456
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    • 1992
  • Expermient were conducted to determine the effect of phosphorus stress on bacteroid content and energy status of soybean (Glycine max [L.] Merr.) nodules. Plants inoculated with Bradyrhizobium japonicum MN 110 were grown with P-stressed (0.05 mM-P) and control (1 mM-P) treatment in the greenhouse. Phosphorus stress decreased nodule mass per plant and nodule mass to whole plant mass ratio. Phosphorus concentration in leaf, stem and root tissues were reduced by 75% but in nodule tissue was reduced only by 40% under phosphorus stress during 3 week experimental period. The bacteroid content per unit nodule mass and the distribution of total nitrogen and total phosphorus among the bacteroid and plant cell fractions of nodule were not affected significantly by phosphorus stress. Regardless of phosphorus treatment, 22% of the nitrogen and 27% of the phosphorus in whole nodules were associated with the bacteroid fraction. The ATP and total adenylate concentrations in and energy charge of whole nodule were decreased 77%, 46% and 37%, respectively, by phosphorus stress. The ATP concentration in and energy charge of the host plant cell fraction of nodules were decreased 86% and 59%, respectively, but these parametres in bacteroid in nodules were not affected by phosphorus stress. These results indicated that nodule is a strong phosphorus sink and that nodule growth and development are regulated to maintain a high phosphorus and energy content in bacteroid even when the host plant is subjected to phosphorus deficiency.

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Effect of Bcl-2 Inhibitor Treatment on Embryo Developmental Competence, Apoptosis and ER-stress in Pigs (Bcl-2의 저해제 처리에 따른 돼지 수정란의 배발달 능력, 세포 사멸 및 소포체 스트레스 양상)

  • Hong, Joo-Hee;Min, Sung-Hun;Lee, Enok;Son, Hyeong-Hoon;Yeon, Ji-Yeong;Park, Humdai;Koo, Deog-Bon
    • Reproductive and Developmental Biology
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    • v.36 no.3
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    • pp.167-172
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    • 2012
  • The key regulators of apoptosis are the interacting protein of the Bcl-2 family. Bcl-2, an important member of this family, blocks cytochrome C release by sequestering pro-apoptotic BH3-only proteins such as Bid, Bad, Bax and Bim. The pro-survival family members (Bcl-2, Bcl-XL, Bcl-W) are critical for cell survival, since loss of any of them causes cell death in certain cell type. However, its role during early porcine embryonic development is not sufficient. In this study, we traced the effects of Bcl-2 inhibitor, ABT-737, on early porcine embryonic development. We also investigated several indicators of developmental potential, including gene expression (apoptosis-related genes) and apoptosis, which are affected by ABT-737. Porcine embryos were cultured in the PZM-3 medium with or without ABT-737 for 6 days. In result, significant differences in developmental potential were detected between the embryos that were cultured with or without ABT-737 ($14.7{\pm}3.0$ vs $30.3{\pm}4.8%$, p<0.05). TUNEL assay showed that the number of containing fragmented DNA at the blastocyst stage increased in the ABT-737 treated group compared with control (4.7 vs 3.7, p<0.05). The mRNA expression of the pro-apoptotic gene Bax increased in ABT-737 treated group (p<0.05), whereas expressions of the anti-apoptotic Bcl-2 family members (Bcl-2, Bcl-XL, Bcl-W) decreased (p<0.05). Also, expressions of the ER stress indicator genes (GRP78, XBP-1 and sXBP-1) increased in ABT-737 treated group (p<0.05). In conclusion, Bcl-2 is closely associated with of apoptosis- and ER stress-related genes expressions and developmental potential in pig embryos.

Production of A Monoclonal Antibody (MAb) Against a Thermal Stable-Soluble Protein in Mackerel and Confirmation of the Properties for the MAb (고등어 어육 중 열안정성 단백질에 특이한 단클론성 항체 개발과 특성 확인)

  • Lee, Jeong-Eun;Kim, Jeong-Sook;Chung, Duck-Hwa;Shim, Won-Bo
    • Journal of Food Hygiene and Safety
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    • v.32 no.1
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    • pp.75-81
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    • 2017
  • For people who have a food allergy the only way to manage the allergy is to avoid the food allergen. The mackerel is one of the major food allergens, but no immunoassay for the rapid and simple detection of mackerel has been reported. The objectives of this study are to develop and characterize monoclonal antibodies (MAbs) specific to mackerel using thermal stable-soluble proteins (TSSP) as an immunogen and to characterize the MAbs by indirect enzyme-linked immunosorbent assay (iELISA). The mice immunized with mackerel TSSP and showing high titer were used for cell fusion and cloning. The characterization of MAbs produced from hybridoma cells obtained was confirmed by indirect ELISA and western blot. Four MAbs were confirmed to be specific to mackerel without cross-reaction to other marine products and livestock products in the both methods. The iELISA and western blot based on the MAbs can sensitively detect 1% mackerel protein in other marine products. These results support that immunochemical methods based on the MAb produced could be used as rapid means to detect low levels of mackerel and to identify mackerel adulterated in food.

Hemopoietic and Radioprotective Effects of Several Herbs on Murine Splenocytes and Bone Marrow Cells (수(數) 종(種) 한약재(韓藥材)가 생쥐의 골수(骨髓) 및 비장세포(脾臟細胞)의 조혈촉진(造血促進)과 방사선(放射線) 방어(防禦)에 미치는 영향)

  • Lee, Neung-Gi;Choe, Seung-Hun;An, Gyu-Seok
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.3 no.1
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    • pp.29-48
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    • 1997
  • Astragalus membranaceus Bunge (黃?), Atractylodes macrocephala Loidz (白朮), Angelica sinensis(Oliv) Kiels (當歸), Lycium chinense Mill (枸杞子), Ligustrum lucidum Ail (女貞子) are Herbs that are frequently used in a lot of prescriptions to reduce the side effects of anti-cancer therapies, especially like chemotherapy and radiotherapy. The radioprotective and hemopoietic effects of these Herbs on BALB/c splenocytes and bone marrow cells are measured. In order to evaluate the Hemopoietic effects, Thymidine uptakes and secretion of colony stimulating factors(CSFs) of splenocytes and myelocytes treated with herbs were measured. Radioprotective effects were accessed by the method of immunocompetence of murine lymphocytes cultured with herbs before irradiation and with LPS, ConA after irradiation. The results are as follows. 1. The orders of Proliferative effects of herbs on splenocytes were Atractylodes macrocephala Loidz (白朮), the higest of all, Astragalus membranaceus Bunge (黃?), Angelica sinensis(Oliv) Kiels (當歸). Lycium chinense Mill (枸杞子) and Ligustrum lucidum Ait (女貞子), the lowest of all. At Optimal concentration, the proliferation ratios of herb-treated splenocytes compared to non-treated ones were like these. Atractylodes macrocephala Loidz (白朮) 44.3, Astragalus membranaceus Bunge (黃?) 17.7, Angelica sinensis(Oliv) Kiels (當歸) 10, Lycium chinense Mill (枸杞子) 6.4, Ligustrum lucidum Ait (女貞子) 2.0.(p<0.05) 2. When splenocytes were cultured during different periods, Atractylodes. macrocephala Loidz (白朮) and Astragalus membranaceus Bunge (黃?) showed the higest proliferation on 3th day, Angelica sinensis(Oliv) Kiels (當歸), Lycium chinense Mill (枸杞子) on 4th day,(p<0.05) and Ligustrum lucidum Ait (女貞子) until 5th day but with no significant increase. 3. The orders of Proliferative effects of herbs on Bone Marrow(BM) cells were Atractylodes macrocephala Loidz (白朮), the higest of all, Astragalus memhranaceus Bunge (黃?), Angelica sinensis(Oliv) Kiels (當歸), Lycium chinense Mill (枸杞子) and Ligustrum lucidum Ait (女貞子), the lowest of all too. At Optimal concentration, the proliferation ratios of herb-treated BM cells compared to non-treated ones were like these. Atractylodes macrocephala Loidz (白朮) 21.7, Astragalus membranaceus Bunge (黃?) 9.9, Angelica sinensis(Oliv) Kiels (當歸) 4.9, Lycium chinense Mill (枸杞子) 2.3, Ligustrum lucidum Ait (女貞子) 1.4(p<0.05). 4. The secretion ratio of colony stimulating factors(CSFs) of each herb-treated group, compared to control, was Atractylodes macrocephala Loidz (白朮) 9.4, Astragalus membranaceus Bunge (黃?) 9.0, Angelica sinensis(Oliv) Kiels (當歸) 4.4, Lycium chinense Mill (枸杞子) 3.8 (p<0.05) but no significant increase in Ligustrum lucidum Ait (女貞子). 5. The mitogen(ConA, LPS) stimulated-lymphocytes cultured with each herb before irradiation of 1-3 Gy showed more enhanced proliferation than control(p<0.05). When compared to each non-irradiated group of all groups, the orders of percentage increase of irradiated group were Atractylodes macrocephala Loidz (白朮), the higest of all, Astragalus membranaceus Bunge (黃?), Angelica sinensis(Oliv) Kiels (當歸), Ligustrum lucidum Ait (女貞子), Lycium chinense Mill (枸杞子). Each percentage showed significant enhancement compared to control group(p<0.05). According to the results, Atractylcdes macrocephala Loidz (白朮), Astragalus membranaceus Bunge (黃?) are suggested to be the most effective hemopoietic and radioprotective herbs, and Angelica sinensis(Oliv) Kiels (當歸), Lycium chinense Mill (枸杞子) the next, but Ligustrum lucidum Ait (女貞子) showed lower effects than expected.

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