• 한국수정란이식학회지
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• 제29권1호
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• pp.1-5
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• 2014

Recent 2 decades, including in vitro maturation (IVM), assisted reproductive technologies (ARTs) achieved noteworthy development. However the efficiency of ARTs with in vitro matured oocytes is still lower than that with in vivo oocytes. To overcome those limitations, many researchers attempted to adapt co-culture system during IVM and consequently maturation efficiency has been increased. The beneficial effects of applying co-culture system is contemplated base on communication and interaction between various somatic cells and oocytes, achievement of paracrine factors, and spatial effects of extracellular matrix (ECM) from somatic cell surface. The understanding of co-culture system can provide some information to narrow the gap between in vitro and in vivo. Here we will review current studies about issues for understanding cu-culture system with various somatic cells to improve in vitro maturation microenvironment and provide bird view and strategies for further studies.

• Reproductive and Developmental Biology
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• 제40권1호
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• pp.7-13
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• 2016

Effectiveness of transgene transfer into genome is crucially concerned in mass production of the bio-pharmaceuticals using genetically modified transgenic animals as a bioreactor. Recently, the mammary gland has been considered as a potential bioreactor for the mass production of the bio-pharmaceuticals, which appears to be capable of appropriate post-translational modifications of recombinant proteins. The mammary gland tissue specific vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals. In this study, to minimize physiological disturbance caused by constitutive over-expression of the exogenous gene, we constructed new retrovirus vector system designed for mammary gland-specific expression of the hEPO gene. Using piggyBac vector system, we designed to express hEPO gene under the control of mammary gland tissue specific and lactogenic hormonal inducible goat ${\beta}$-casein or mouse Whey Acidic Protein (mWAP) promoter. Inducible expression of the hEPO gene was confirmed using RT-PCR and ELISA in the mouse mammary gland cells treated with lactogenic hormone. We expect the vector system may optimize production efficiency of transgenic animal and reduce the risk of global expression of transgene.

• Clinical and Experimental Reproductive Medicine
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• 제25권1호
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• pp.77-86
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• 1998

The uptake of glucose for metabolism and growth is essential to most animal cells and is mediated by glucose-transporter (GLUT) proteins. The aim of this study was to determine which class of glucose transporter molecules was responsible for uptake of glucose in the mouse early embryo and at which stage the corresponding genes were expressed. In addition, co-culture system with vero cell was used to investigate the effect of the system on GLUT expression. Two-cell stage embryos were collected from the superovulated ICR female and divided into 3 groups. As a control, embryos were cultured in 0.4% BSA-T6 medium which includes glucose. For the experimental groups, embryos were cultured in either co-culture system with vero cells or glucose-free T6 medium supplemented with 0.4% BSA and pyruvate as an energy substrate. 2-cell to blastocyst stage embryos in those groups were respectively collected into microtubes (50 embryos/tube). Total RNA was extracted and RT-PCR was performed. The products were analysed after staining ethidium bromide by 2% agarose gel electrophoresis. Blastocysts were collected from each group at l20hr after hCG injection. They were fixed in 2.5% glutaraldehyde, stained with hoechst, and mounted for observation. In control, GLUT1 was expressed from 4-cell to blastocyst. GLUT2 and GLUT3 were expressed in morula and blastocyst. GLUT4 was expressed in all stages. When embryos were cultured in glucose-free medium, no significant difference was shown in the expression of GLUT1, 2 and 3, compared to control. However GLUT4 was not expressed until morular stage. When embryos were co-cultured with vero cell, there was no significant difference in the expression of GLUT1, 2, 3 and 4 compared to control. To determine cell growth of embryos, the average cell number of blastocyst was counted. The cell number of co-culture ($93.8{\pm}3.1$, n=35) is significantly higher than that of control and glucose-free group ($76.6{\pm}3.8$, n=35 and $68.2{\pm}4.3$, n=30). This study shows that the GLUT genes are expressed differently according to embryo stage. GLUTs were detectable throughout mouse preimplantation development in control and co-culture groups. However, GLUT4 was not detected from 2- to 8-cell stage but detected from morula stage in glucose-free medium, suggested that GLUT genes are expressed autocrinally in the embryo regardless of the presence of glucose as an energy substrate. In addition, co-culture system can increase the cell count of blastocyst but not improve the expression of GLUT. In conclusion, expression of GLUT is dependent on embryo stage in preimplantation embryo development.

• 한국식품위생안전성학회지
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• 제31권2호
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• pp.113-118
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• 2016

Proprotein convertase subtilisin/kexin type 9 (PCSK9) 유전자는 주로 간에서 분비되며 저밀도지단백질 수용체(LDLR)와 결합체를 이루어 세포 내 저밀도지단백질 수용체를 약화시킴에 따라 저밀도지단백질(LDL) 흡수를 감소시킨다. PCSK9은 심혈관 질환 및 상염색체 우성 가족성 고콜레스테롤혈증과 깊은 연관성을 나타내며 발병 메커니즘을 조절하는 주요한 인자로서 치료제 개발을 위해 많은 연구가 진행 중에 있다. 하지만 생식계통 내에서 PCSK9의 발현 및 기능에 대한 연구가 미비한 현실이다. 이에 본 연구에서는 고 콜레스테롤 식이에 따라 수컷 랫드의 생식기관 (음경, 전립선, 정소) 내 Pcsk9의 발현 변화를 확인하고자 한다. 과도한 콜레스테롤의 축적으로 인한 이상지질혈증은 수컷 랫드의 생식기능장애와 내분비교란을 유발하지만 이를 일으키는 조절 인자와 메커니즘은 명확하게 밝혀지지 않았다. 본 연구진의 선행연구에서는 고 콜레스테롤 식이에 의해 혈중 총 콜레스테롤, 저밀도지단백질 콜레스테롤 레벨이 증가 및 고밀도지단백질(HDL) 콜레스테롤의 감소를 확인하였다. 이 후, Pcsk9의 mRNA 발현은 음경 조직 내에서 80% 감소하는 것으로 나타났고 그 발현은 주로 발기 조직인 음경해면체 및 요도해면체 내 요도상피세포에서 주를 이룸을 확인하였다. 또한 Pcsk9은 고콜레스테롤 식이에 의해 발생하는 전립선 비대증 조직 내에서 20% 감소하는 것으로 나타났다. 그리고 마지막으로 Pcsk9은 고 콜레스테롤 식이를 섭취한 랫드의 정소 조직내에서 정상 정소 조직에 비해 약 28% 감소하였으며 정자발생에 주요한 정세관 내 정원세포, 정모세포, 정세포에서 그 발현이 확인되었다. 본 연구결과를 통해 Pcsk9 유전자가 수컷 랫드 생식기관 (음경, 전립선, 정소)에서 고콜레스테롤 식이에 의해 발현이 감소하는 것을 확인하였으며, 이는 Pcsk9이 남성 생식기관의 성장 및 발달 메커니즘에 중요한 역할을 미칠 것으로 사료된다.

• Journal of Microbiology and Biotechnology
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• 제10권3호
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• pp.281-286
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• 2000

2,3,7,8-Tetrachlorodibenzo-p-Dioxin (TCDD), a ubiquitous environmental contaminant, causes a variety of adverse effects on the male reproductive system in rats. The effect of green tea extract (GTE) was investigated on the testicular function in Spragure-Dawley rats after a single exposure of 10$\mu\textrm{g}$ TCDD/kg body weight. The exposure of rat to TCDD significantly increased the weights of the epididymis and ventral prostate, yet significantly decresed the weight of the seminal vesicle when compared to the controls (p<0.05). In a combined treatment of TCDD with GTE, the organ weight changes caused by TCDD treatment disappeared. Significant decreases in sperm motility and sperm numbers were observed in the TCDD-treated rats, when compared to the control (p<0.05). GTE treatment reversed the decrease of sperm motility and sperm numbers caused by TCDD. There were no differences in sperm morphology, histological changes of the reproductive organs, and spermatogenesis between all the treated groups. In the ventral prostate and seminal vesicle, TCDD increased the CYP1A1 mRNA level, however, it did not affect the estrogen receptor $\beta$ (ER-$\beta$) mRNA level. GTE treatment did not influence the effect of TCDD on the levels of CYP1A1 and Er-$\beta$ mRNA. These results seem to indicate that green tea protects the testicular function against TCDD-induced reproductive toxicity, not because of a receptor-mediated mechanism but rather due to a secondary change of testes or accessory sex organs.

• 한국산업보건학회지
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• 제18권3호
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• pp.224-238
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• 2008

The aim of this study is to confirm the physicochemical property and hazard of thinner (012), which is a diluent of enamel paint used for floor coating for waterproofing and oil painting for the outer wall. The literatures of physicochemical property and hazard of thinner were surveyed and its physicochemical property were evaluated. And then, the inhalation toxicity of thinner affecting the central nervous system and reproductive organs in rats were examined by subchronic (6 h./day. 5 days/ week for 13 weeks) inhalation test. 1) According to the 13-week subchronic inhalation test, there were no significant changes in clinical test and body weight. However, a significant evidence of toxicity was observed in the hematological test and organ weight such as heart, kidney, liver and brain (p<0.01) in the 200 ppm and 1,000 ppm exposure groups in a dose response manner. In the histopathology analysis, there were no significant evidence of toxicity. Therefore, thinner was not classified as an organ targeted toxic agent. In case of Harmfulness, it could be classified as a chronic toxic agent 3($500 ppm/4hr, rat). 2) The reproductive toxicity such as extension of the period of estrous cycle, reduction of serum estradiol concentration and increase of frequency of the abnormal sperm was observed in the 1,000 ppm exposed animals. 3) The result of the physicochemical property of the test material showed that the specific gravity was 0.793, boiling point $155.8^{\circ}C$, steam pressure 2.1 kPa, ignition point $34.5^{\circ}C$, and spontaneous ignition point $280^{\circ}C$. The endothermic and exothermic values were 371.4 J/g and 159.1 J/g. respectively. The explosion limit was 214 mg/l. These data showed that thinner could be classified as an explosion agent level 1.2 and ignitive liquid agent 3 ($23-60^{\circ}C$) according to the notification No. 2008-1 of the Labor Ministry, "Classifying Standard of Chemical Materials."

• 한국작물학회지
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• 제27권4호
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• pp.361-370
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• 1982

The effects of climatic factors on organisms lire variable and complex, and it, however, can be interpreted in terms of those on the distribution and those on the population densities. The distribution of an organism may largely be determined by the temperatures, except some temporal organisms which are depended on the air mass movements. Population density of an organism is determined by various climatic factors, such as previous winter temperature, temperature of growing season and rainfall. The start of growing season of the rice plants has been shifted to earlier since last decade in Korea. This may mean that the overall climatic condition during the growing season might be considerably different from those in past years, and such a difference in climatic conditions might have close relation with the recent status of the diseases and insect pests through direct effects on the physiology and population dynamics of the organisms, as well as through on the biotic associations of the pest organisms. The white back planthopper and brown planthopper have become the key insect pests in Korea in recent years. They are migratory and have high reproductive pontentials and more generations than average residential insects. The synchronization of the migrants and physiological condition of the rice plants seems to be the important factors in relation to the recent outbreaks of these insects; the high reproductive rate can be obtained with the growth stage of rice being 30-50 days after transplanting. The modication of the microclimate associated with high plant density and some other introduced new cultural techniques also have some relation with the outbreak. The key diseases of the rice are the blast disease, sheath blight and the bacterial leaf blight. For the rice blast, the seedling blast and leaf blast during the early growing season and the neck blast, have become more serious, the former may be related to hotbed nursery and the later may be related to the high humidity in early August, and synchronization of the heading time which has been shifted to early part from middle or late part of August. In general, for the rice diseases, the development of the new races have been the most serious which are largely resulted from the introduction of the new varieties, but it also seems to be related with the prolonged periods of the favorable condition associated with the shifted growing seasons. In general, the diseases and insect pest problems have become much more variable and complex, and control measures should be based on the thorough knowledge of the ecology of the pest organisms, that is, effects of various environmental factors on the disease cycle; spore release, spore deposition, infection, colonization and sporulation of the disease organisms, and those on the development, reproductive potentials, dispersal, age specific responses of the insects. The well organized real-time pest management systems, such as alfalfa weevil management system developed at the Purdue University in U.S., is the prime importance for the implementation of the pest management principles.

• Clinical and Experimental Reproductive Medicine
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• 제38권4호
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• pp.193-202
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• 2011

Objective: We found previously that $interferon$ $regulatory$ factor ($Irf$)-1 is a germinal vesicle (GV)-selective gene that highly expressed in GV as compared to metaphase II oocytes. To our knowledge, the function of $Irf-1$ in oocytes has yet to be examined. The present study was conducted to determine the relationship between retinoic acid (RA) and RA-mediated expression of $Irf-1$ and the mouse oocyte maturation. Methods: Immature cumulus-oocyte-complexes (COCs) were collected from 17-day-old female mice and cultured $in$ $vitro$ for 16 hours in the presence of varying concentrations of RA (0-10 ${\mu}M$). Rate of oocyte maturation and activation was measured. Gene expression was measured by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) and cytokine secretion in the medium was measured by Bio-Plex analysis. Apoptosis was analyzed by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Results: The rates of oocyte maturation to metaphase II and oocyte activation increased significantly with RA treatment (10 nM-1 ${\mu}M$). With 100 nM RA treatment, lowest level of $Irf-1$ mRNA and cumulus cell's apoptosis was found. Among 23 cytokines measured by Bio-Plex system, the substantial changes in secretion of tumor necrosis factor-${\alpha}$, macrophage inflammatory protein-$1{\beta}$, eotaxin and interleukin-12 (p40) from COCs in response to RA were detected. Conclusion: We concluded that the maturation of oocytes and $Irf-1$ expression are negatively correlated, and RA enhances the developmental competence of mouse immature oocytes $in$ $vitro$ by suppressing apoptosis of cumulus cells. Using a mouse model, results of the present study provide insights into improved culture conditions for $in$ $vitro$ oocyte maturation and relevant cytokine production and secretion in assisted reproductive technology.

• 한국패류학회지
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• 제27권3호
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• pp.181-190
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• 2011

본 연구는 백합과 이매패류인 바지락, Ruditapes philippinarum을 노닐페놀에 노출시켰을 때, 생식 및 조직학적 지표 변화에 대해서 조사하였다. 실험기간은 16주였다. 실험구는 1개의 대조구와 3개의 노닐페놀 노출구인 0.05, 0.1, $0.2 mg\;NP\;L^{-1}$였다. 생식소지수는 암컷의 경우, 대조구보다 노출구에서 더 높았으며, 수컷의 경우, 대조구보다 노출구에서 더 낮았다. 이성생식세포 발현율은 14.56%였으며, 수컷에서의 발현율이 더 높았다. 기관계 가운데 외투막에서 혈림프동의 확장, 내부상피층 자유면에 발달하는 선조연의 탈락과 박리, 점액세포의 변성이 관찰되었다. 아가미에서는 정단섬모와 측면섬모의 부분적 소실, interlamella junction 붕괴, 점액세포의 변성이 확인되었다. 발에서는 상피세포의 증식, 선조연의 소실 및 섬모가 탈락되었다. 지방갈색소 축적은 노닐페놀에 농도 의존적으로 높았다. 본 연구결과 노닐페놀은 바지락의 낮은 생존율과 조직병리학적 변화 및 이성생식세포 발현을 유도하며, 더욱이 생식과 관련하여 estrogenic effector로서 가능성을 확인하였다.

• 한국동물위생학회지
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• 제42권4호
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• pp.201-216
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• 2019

Porcine reproductive and respiratory syndrome (PRRS) is characterized by reproductive failure in sows and respiratory distress in all age pigs. Porcine respiratory disease complex (PRDC) is a disease caused by opportunistic bacterial infection secondary to a weakened immune system by a preceding respiratory infection. In this study, we tried to compare the immune responses in PRRS and PRDC groups to clearly characterize the disease severity. Eighty-five pigs were infected with various Korean field PRRS virus strains. Infected animals were classified into PRRS (n=32) and PRDC (n=53) groups based on lung lesions such as interstitial pneumonia, suppurative pneumonia, and pleuropneumonia. The immune cell population of bronchoalveolar lavage cells (BALc) was evaluated on 14 and 28 days post infection (dpi) and PMBC cytokine expression was measured on 0, 3, 7, 14 dpi to investigate early inflammatory reactions. Pulmonary lesion severity was negatively correlated with alveolar macrophage (AM) in both PRRS and PRDC groups on 14 and 28 dpi. AM in BALc was less populated in PRDC group on 28 dpi compared to PRRS group. AM in BALc was significantly less populated in PRDC group on 28 dpi compared to 14 dpi. In addition, cytotoxic T lymphocyte (CTL) in BALc was higher populated in PRDC group on 14 dpi and 28 dpi compared to PRRS group. In the case of PBMC cytokine TNF-α, IFN-α, IL-1β, IFN-γ, FoxP3, and IL-2, the PRRS group showed higher expression than the PRDC group on 7 dpi, 14 dpi, 7 dpi, 14 dpi, 14 dpi, and 14 dpi, respectively. On the other hand, in the case of IFN-β, IL-6, IL-8, IL-4, and IL-17, the PRDC group showed higher PBMC cytokine expression at 14 dpi, 7 dpi, 14 dpi, 3 dpi, and 3 dpi, respectively, than the PRRS group. Based on these results, our study could characterize differential immune responses in pigs with PRRS or PRDC.