• Asian-Australasian Journal of Animal Sciences
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• 제26권3호
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• pp.334-342
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• 2013

Proteases and protease inhibitors play key roles in most physiological processes, including cell migration, cell signaling, and cell surface and tissue remodeling. Among these, the matrix metalloproteinase (MMPs) pathway is one of the most efficient biosynthetic pathways for controlling the activation of enzymes responsible for protein degradation. This also indicates the association of MMPs with the maturation of spermatozoa. In an attempt to investigate the effect of MMP activation and inhibitors in cultures with various hormones during sperm capacitation, we examined and monitored the localization and expression of MMPs (MMP-2 and MMP-9), tissue inhibitors of metalloproteinases (TIMP-2 and TIMP-3), as well as their expression profiles. Matured spermatozoa were collected from cultures with follicle-stimulating hormone (FSH), luteinizing hormone (LH), and Lutalyse at 1 h, 6 h, 18 h, and 24 h. ELISA detected the expression of MMP-2, MMP-9, TIMP-2, and TIMP-3 in all culture media, regardless of medium type (FSH-supplemented fertilization Brackett-Oliphant medium (FFBO), LH-supplemented FBO (LFBO), or Lutalyse-supplemented FBO (LuFBO)). TIMP-2 and TIMP-3 expression patterns decreased in LFBO and LuFBO. MMP-2 and MMP-9 activity in FBO and FFBO progressively increased from 1 h to 24 h but was not detected in LFBO and LuFBO. The localization and expression of TIMP-2 and TIMP-3 in sperm heads was also measured by immunofluorescence analysis. However, MMPs were not detected in the sperm heads. MMP and TIMP expression patterns differed according to the effect of various hormones. These findings suggest that MMPs have a role in sperm viability during capacitation. In conjunction with hormones, MMPs play a role in maintaining capacitation and fertilization by controlling extracellular matrix inhibitors of sperm.

• Asian-Australasian Journal of Animal Sciences
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• 제23권3호
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• pp.326-332
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• 2010

Daidzein, a natural isoflavonoid phytoestrogen, structurally resembles estradiol (E2) and possesses estrogenic activity. This study was designed to test the hypothesis that daidzein may mimic the effects of E2 on ovine follicle development by regulation of the mRNA expression of bone morphogenetic protein receptor genes and thereby influence the reproductive system. Granulosa cells were cultured in serum-free McCoy's 5A medium with and without supplementation of daidzein. Results showed that daidzein (10-100 ng/ml) significantly increased the proliferation of ovine granulosa cells (p<0.05), but inhibited the growth of granulosa cells at a dose of 1,000 ng/ml (p<0.01). Daidzein inhibited progesterone production in a dose dependent manner; however, it did not affect estradiol production by granulosa cells. We also investigated the effects of daidzein on BMPRII, BMPRIB and ALK-5 mRNA expression in ovine granulosa cells by quantitative real-time PCR. Treatment of granulosa cells with daidzein increased significantly expression of these genes at 10-100 ng/ml. Thus, these data suggested that a low concentration of daidzein (10-100 ng/ml) had a direct stimulatory effect on ovine granulosa cells while a high concentration was toxic.

• Asian-Australasian Journal of Animal Sciences
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• 제27권11호
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• pp.1526-1531
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• 2014

Matrix metalloproteinases (MMP) are key enzymes involved in cell and tissue remodeling during ovarian follicle development and ovulation. The control of MMP9 transcription in ovarian follicles occurs through a core promoter region (-2,400 to -1,700 bp). The aim of this study was to screen genetic variations in the core promoter region and examine MMP9 transcription regulation and reproduction performance. A single cytosine deletion/insertion polymorphism was found at -1954 $C^+/C^-$. Genetic association analysis indicated significant correlation between the deletion genotype ($C^-$) with total egg numbers at 28 weeks (p = 0.031). Furthermore, luciferase-reporter assay showed the deletion genotype ($C^-$) had significantly lower promoter activity than the insertion genotype ($C^+$) in primary granulosa cells (p<0.01). Therefore, the identified polymorphism could be used for marker-assisted selection to improve chicken laying performance.

• 한국발생생물학회지:발생과생식
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• 제17권2호
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• pp.141-147
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• 2013

Keeping the intact germinal vesicle (GV) is essential for maintaining the capacity of mammals including human. It is maintained by very complex procedures along with folliculogenesis and is a critical step for getting competent oocyte. So far, a few mechanisms involved in folliculogenesis are known but GV arrest mechanisms are largely unrevealed. Cyclic AMP, a adenosine derived substance, have been used as inhibitor of germinal vesicle breakdown as a putative oocyte maturation inhibitor. In this study, we examined the potency of adenosine as GV maintainer and a possible signaling mediator for that. A1, A2b, and A3 were detected in cumulus cells of cumulus enclosed-oocyte (CEO). Intact of germinal vesicle was not kept like in follicle but the spontaneous maturation was inhibited by exogenous adenosine. It is inhibited with concentration dependent manners. Intracellular calcium level of cumulus was extensively increased after adenosine treatment. Based on these results it is suggested that one of the pathway for GV arrest by adenosine and its receptors is calcium mediated signaling pathway in CEO.

• 대한수의학회지
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• 제41권3호
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• pp.373-380
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• 2001

Marek's disease virus (MDV) can cause skin lesions including inflammatory to tumorous. The phenotypical changes of lymphocytes infiltrating in the skin lesions induced by MDV were not clear. Therefore, the skin biopsies taken at weekly intervals for 8 weeks from the same specific-pathogen free chickens inoculated with Md/5 MDV were examined to analysis the phenotypical changes of lymphocytes. Histologically skin lesions progressed from initial inflammatory to late tumorous. Sequentially CD4+ T lymphocytes increased gradually in number from initial skin lesions and were major composition cells in the tumor lesions. Regardless of inflammatory or tumor lesions, CD8+ T cells and ${\gamma}{\delta}$ T cells infiltrated particularly in the dermis and subcutaneous on which MDV was actively replicated in the feather follicle epithelium(FFE). In addition, IgG bearing B lymphocytes in considerable number infiltrated in the dermis and subcutaneous tissues. From these results, the development of MDV-induced skin lesions was inflammatory following tumorous. In addition, each CD8+, ${\gamma}{\delta}$ and CD4+ T cells and B cell might act to protect MDV replication in the FFE or tumor cells which turned on lytic cycle.

• 한국수정란이식학회지
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• 제11권3호
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• pp.241-248
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• 1996

This study was conducted to improve the production efficiency of in vitro produced (IVP) embryos in Korean Native cows. The optimal conditions and procedures for in vitro maturation(IVM), in vitro fertilization(IVF) and in vitro culture(IVC) of bovine follicular oocytes and IVP embryos were evaluated. Immature follicular oocytes were collected fiom the follicles of bovine ovaries obtained from abattoirs. The oocytes of Grade I and II for IVM were cocultured with monolayered bovine oviductal epithelial cells(BOEG) or granulosa cells in TCM-199 solution supplemented with follicle stimulating hormone, lutenizing hormone, estradiol-17$\beta$ and heat inactivated fetal calf serum at 39$^{\circ}C$ under 5% $CO_2$ in air for 14 to 24 hours. Most of the oocytes(93%) matured to metaphase II in 24 hours. The cocultured IVM oocytes were fertilized in vitro at significantly(P<0.05) higher rate with BOEC(83.8%) and with granulosa cells(84.6%) than the non-cocultured IVM oocytes(73.6%). The IVM-IVF embryos developed to morula and blastocyst at significantly(P<0.05) higher rate in coculture with BOEC(41.2%) than with granulosa cells(23.1%) or conditioned medium(23.4%).

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.78-78
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• 2001

본 연구는 초자화동결법(vitrification)을 이용하여 미성숙 소 난포란의 동결기술을 개발하기 위하여 시도하였으며 난포의 크기와 동결보호제의 농도 및 난황의 첨가가 동결-융해 후 미성숙 소 난포란의 생존성에 미치는 영향을 조사하였다. 미성숙 난자는 3mm 미만과 3mm 이상으로 구분된 난포로부터 채란하였으며 동결처리 전 동결보호제로서 30%EG과 40%EG, 그리고 각각에 10%의 egg yolk을 첨가하여 동결-융해 후 미성숙 난자의 생존성과 체외성숙율 및 체외수정율을 비교하였다. 생존율과 성숙율에 있어서 3m 미만과 3mm 이상의 2 처리구 모두 40%EG에 10%의 egg yolk을 첨가할 경우 유의적으로 높은 결과를 보였으나 3mm이하에서는 EG수준과 egg yolk 첨가 유무간에 생존율과 성숙율에 유의적인 차이가 나타나지 않았다. 동결-융해 후 체외수정율은 3mm 미만에서는 처리구간에 유의적인 차이를 보이지 않았으나 3mm이상의 경우에서는 40%EG에 10%의 egg yolk 첨가구가 다른 처리구에 비해 유의적으로 높았다. 난포의 크기에 따른 동결-융해 후 생존율은 3mm미만의 난포란이 3mm이상 보다 높았으나 성숙율과 수정율에 있어서는 3mm이상이 3mm미만의 난포란에서 보다 다소 높았다. 생존율, 성숙율 및 수정율 모두 난포크기간에 유의차는 인정되지 않았다.

• Reproductive and Developmental Biology
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• 제38권2호
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• pp.63-70
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• 2014

Plasminogen activators (PAs) are serine proteases that convert plasminogen to plasmin. Two type of PAs are urokinase-type PA (uPA) and tissue-type PA (tPA). Plasminogen is present in most extracellular fluids. PAs play in various reproductive processes including implantation, ovulation and fertilization. In the spermatozoa, PAs and PAIs play a role in sperm motility and fertilization. PAs in the sertoli cell are stimulated spermatozoa maturation and sperm activation through the phospholipase A2. The oocyte maturation is the process for fertilization and implantation. PAs in cumulus-oocyte complexes (COCs) are related to oocyte maturation by protein kinase A and C. In the ovulatory process, PAs activity are changed and it are related to reducing the tensile strength of ovarian follicle wall. The uterine environment is important for reproduction and the uterus undergo tissue remodeling. In the uterus and oviduct of mammals, expression and activity of PAs are changed during estrous cycle. Thus, expression and activity of PAs are concerned to many reproductive functions. Therefore, PAs seem to important factor of regulator in reproductive events.

• Clinical and Experimental Reproductive Medicine
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• 제16권1호
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• pp.35-40
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• 1989

In order to study the follicular atretic mechanism in mammalian ovary, the relationship between pyknotic index (PI) of granulosa cells and the steroid concentrations in the follicular fluid of atretic follicle was investigated. Follicles were isolated from porcine ovary according to their sizes and the reproductive phases. Steroid concentrations were quantified by high-performance liquid chromatography. As PI increased, the concentration of progesterone was significantly increased(p<0.05), whereas testosterone and estradiol showed no significant changes in their concentration. As follicular size was increased, PI of follicular GC in the luteal phase was increased significantly(p<0.05) and the molar ratio of progesterone to testosterone was increased in the follicles of follicular phase. It can be concluded that progesterone accumulated in the follicular fluid as atresia of the follicles was progressed, and that PI of granulosa cells could be used as one of convenient and pratical criteria for the identification of follicular atresia.

• 한국임상약학회지
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• 제26권3호
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• pp.187-194
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• 2016

Background: Alopecia areata (AA) is a very disturbing and expensive disorder in which the exact etiology is not known and it is yet to be treated completely well. Most alopecia patients exhibit some inflammation in the hair follicles regardless of the causes. The clinical symptoms of alopecia present very diversely while the prime symptom is local intermittent fever which are related to inflamed cells. Methods: This study aimed to evaluate how repetitive intermittent fever can damage the normal human dermal fibroblast (NHDF) cells and investigated the cytotoxic and proliferative effects after application of new candidate drugs (ibuprofen, menthol, cetirizine) for alopecia in comparison to minoxidil. Results: This study demonstrated that ibuprofen, menthol, or/and cetirizine can prevent or slow down the damage of NHDF cells from intermittent fever in early alopecia. Aggressive preventative intervention with those drugs before complete destruction of hair follicle by excessive repetitive fever, is a very important step for alopecia therapy and these drugs are recommended as candidate drugs for alopecia in the future. Conclusion: Aggressive preventative intervention with drugs before complete destruction of hair follicles (NHDF cells) by excessive repetitive fever is a very important step for alopecia therapy or progression.