• Title/Summary/Keyword: Reproductive tract

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The gene encoding guanidinoacetate methyltransferase (GAMT) maps to mouse chromosome 10 near the locus of hesitant mutation affecting male fertility

  • Chae, Young-Jin;Chung, Chan-Ee;Kim, Byung-Jin;Lee, Mun-Han;Lee, Hang
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 1998.07a
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    • pp.50-51
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    • 1998
  • guanidinoacetate methyltransferase (GAMT) catalyzes the last step of creatine biosynthesis in mammals. Creatine plays an important role in cellular energy metabolism in variety of tissues including brain and male reproductive tract. Congenital deficiency of the enzyme leads to a neurologic disorder in humans. We used an interspecific backcross DNA panel to map Gamt to the central region of mouse Chromosome (Chr) 10 near the locus of hesitant mutation affecting male fertility. We assigned the human GAMT gene to Chr 19 by PCR analysis of a human/rodent somatic hybrid cell line DNA panel, and further localized the human gene to Chr 19 at band p13.3 by PCR analysis of a human radiation hybrid DNA panel. Human chr 19p13.3 is homologous to the central part of mouse Chr 10 where mouse Gamt is located. Furthermore, this part of mouse Chr 10 contains mutant loci the phenotype of which is similar to the GAMT deficiency in human.

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Embryo transfer in Korean Native Black Goat: Embryo recovery and transfer for the production of transgenic goat (한국재래흑염소 수정란의 이식: 형질전환 흑염소 생산을 위한 수정란의 채취와 이식)

  • 신상태
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2000.06a
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    • pp.64-75
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    • 2000
  • During the last three decades considerable advances has been made in goat embryo production and transfer technology. The Korean native black goat is the most useful domestic ruminant in this country for biological investigation and application because it has a lot of merits such as relatively short generation period(1 vs 2 year for a cow), easy of handling, well adaptation, high fertility, convenient and inexpensive. This article covers the methods of superovulation, estrus synchronization, embryo collection and transfer techniques, pregnancy diagnosis and subsequent pregnancy and kidding rates for the production of transgenic Korean native black goats. More than one hundred goat kids have been produced as a result of our transgenic goat project via microinjection of foreign gene into pronuclei, in vitro culture, transfer of various stages of fresh and frozen-thawed microinjected embryos into oviducts or uteri of recipient does. We have got two transgenic goats carrying a transgene targeting the expression of recombinant human granulocyte colony stimulating factor(hG-CSF) to the mammary gland so far. Since collection and transfer of embryos in this species is usually accomplished by laparotomy, exteriorization of the reproductive tract for surgical embryo collection leads to the formation of post-operative adhesions. Nonsurgical or laparoscopic technique to reduce adhesions from repeated surgeries has great advantages in improving embryo production and transfer especially from valuable donors. We will discuss this later.

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Serological and bacteriological study on canine brucellosis in the large kennel farms in Gyeongbuk province (경북지방 개 브루셀라병 다두 발생농장의 혈청학적 및 세균학적 조사)

  • Kim, Seong-Guk;Seo, Hee-Jin;Kim, Soon-Tae;Jang, Young-Sul;Jo, Min-Hee
    • Korean Journal of Veterinary Service
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    • v.33 no.2
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    • pp.129-134
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    • 2010
  • Canine brucellosis is a contagious disease of the reproductive tract that cause mainly abortion and infertility in dog. A serological and bacteriological survey was conducted for breeding kennels which were suffered from frequent outbreak of canine brucellosis in Gyeongbuk province in 2009. Among 138 samples, 45 serum samples were sero-positive. Brucella canis was isolated from 30 blood samples of the seropositive cases, and from 2 samples of 62 sero-negatives. The biochemical properties of 32 isolates were characterized with no production of H2S, no fermentation of carbohydrates, hydrolyzation of urea, and development of thionin dye medium. At amplification of BCSP and 16S-rRNA gene using PCR, 711bp and 905bp DNA fragments were detected in agarose. Three tandem repeat pattern was shown in genotyping by Multi-locus VNTR assay (MLVA).

The Effect of GM-CSF Supplementation in Culture Medium in the Human IVF Programs (체외수정 시술시 배양액에 첨가된 과립구 대식세포 증식인자 (Granulocyte-Macrophage Colony Stimulating Factor)의 효과)

  • Park, Won-Il;Kwon, Hynck-Chan;Kim, Dong-Hoon;Kang, Hee-Kyoo;Kim, Myo-Kyung;Lee, Hoi-Chang;Jung, Ji-Hak;Lee, Myong-Seop;Lee, Ho-Joon
    • Clinical and Experimental Reproductive Medicine
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    • v.28 no.2
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    • pp.161-167
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    • 2001
  • Objective: Granulocyte-macrophage colony stimulating factors known to be secreted in murine and human reproductive tract. The development of human, bovine and murine embryos could be promoted by addition of GM-CSF in culture medium. However, the pregnancy and implantation rate of embryos cultured in GM-CSF have not been evaluated. The aim of this study was to assess the effect of GM-CSF in embryo development, pregnancy and implantation rate. Methods: A total of 191 IVF cycles were divided into control and GM-CSF supplement group (control=96, GM-CSF=95). The embryos were cultured for three day with or without 2 ng/ml of recombinant human GM-CSF. The quality of embryo, developmental velocity, pregnancy and implantation rates were compared. Results: There was no difference in age, number of gonadotropin ampules used, number of oocytes and fertilization. The number of ICSI cycle was higher in GM-CSF group. In GM-CSF group, G-1 grade embryos were the highest in proportion (56.4%), while G-2 grade embryos were highest (44.3%) in control group. The developmental velocity of embryos were not different between GM-CSF and control group. The pregnancy and implantation rates were significantly higher in GM-CSF group than control (47.4% vs. 33.3%, 17.0% vs. 11.1% respectively). Conclusion: By adding GM-CSF in culture medium, the quality of embryo, pregnancy and implantation rate could be improved.

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The Effects of Flutamide and Diethylstilbestrol Exposure on the Reproductive Organs and Thyroid of Male Rats by the Rodent 20-day Thyroid/Pubertal Assay

  • Moon, Hyun-Ju;Shin, Jae-Ho;Kim, Hyung-Sik;Lee, Su-Jung;Kang, Il-Hyun;Kim, Tae-Sung;Seok, Ji-Hyun;Kim, In-Young;Han, Soon-Young
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2002.10a
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    • pp.168-168
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    • 2002
  • To establish a test protocol for the rodent 20-day thyroid/pubertal assay, flutamide and diethylstilbestrol (DES) were administered to intact male Sprague-Dawley rats from postnatal day 33 for 20 days. Flutamide (1, 5, and 25 mg/kg/day) or DES (10, 20, and 40 ug/kg/day) was given once daily by oral gavage to immature male rats. Prepuce separation was significantly delayed in flutamide group and in DES group. One day after the last dose, the rats were killed and pituitary, thyroid, and reproductive organs were removed and weighed. Flutamide treatment resulted in a significant reduction in the weights of epididymides, ventral prostate, seminal vesicles plus coagulating glands and fluid (SVCGF), levator ani. bulbocarvenus muscles (LABC), Cowper's glands, and glans penis. The weight of adrenal glands decreased at % mg/kg/day, while testes and any other organ weights were unaffected. No microscopic changes were observed in the thyroid glands. Serum levels of testosterone wert significantly increased in the flutamide-treated groups and serum levels of estradiol were also increased. A significant reduction in the weights of testes, epididymides, ventral prostate, SVCGF, LABC, Cowpers glands, and glans penis of DES treated group. Serum testosterone and LH decreased significantly in DES group. Decrease of estradiol was observed, but not significant. These results indicate that flutamide and DES delay puberty in the male rat and its mode of action appears to be via altered secretion of steroids, which subsequently affect the development of the reproductive tract. (Supported by the grant from NITR/Korea FDA for Endocrine Disrupter Research.)

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Effects of an Endocrine Disruptor (Bisphenol A) on the Mouse Spermatogenesis (내분비계 장애물질 Bisphenol A가 생쥐의 정자형성에 미치는 영향)

  • Nam, Hyun-Sik;Seo, Dong-Sam;Ko, Yong
    • Development and Reproduction
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    • v.4 no.1
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    • pp.53-59
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    • 2000
  • Endocrine disruptors (EDs) are exogenous chemicals which interfere several aspects of natural hormone properties. EDs with estrogenic activity have been recently reported to cause animal reproductive problems. This study was performed to investigate the effects of bisphenol A (BPA) on the mouse spermatogenesis in vivo. Male ICR mice were orally injected on a daily basis with low dose of BPA 20 mg/kg, high dose of BPA 200 mg/kg, or corn oil (vehicle control) for 7 days, and litter size and weights of body, testis, and cauda epididymis were measured. The level of serum testosterone and the expression of TGF- $\beta$$_1$ mRNA were also analyzed using RIA and RT-PCR, respectively. Also, morphological differences of testes after treatments were examined. Sperm concentration and level of serum testosterone showed a decreasing tendency detected as untreated >corn oil >low >high dose BPA treated mice, although there were no significant statistical differences. Interestingly, in mice treated with a high dose of BPA, partial disappearance of spermatozoa in seminiferous tubular lumen and the expression of TGF-$\beta$$_1$ mRNA were observed. Spermatogenesis was disrupted through TGF-$\beta$ system in the seminiferous tubules, resulting in no development of germ cells. Similarly, the litter size treated with a high dose of BPA was significantly different from that of untreated control group. In conclusion, these results that a high dose of BPA (200 mg/kg) acts as an endocrine disruptor during apermatogenesis in male mice md that there are BPA-specific lesions in the adult male reproductive tract might represent a permanently altered responsiveness to testosterone by BPA in the affected target tissue.

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Polydnavirus Replication and Ovipositional Habit of Cotesia plutellae (프루텔고치벌(Cotesia plutellae) 폴리드나바이러스 복제와 산란 습성)

  • Kim Yonggyun;Bae Sangki;Lee Sunyoung
    • Korean journal of applied entomology
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    • v.43 no.3 s.136
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    • pp.225-231
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    • 2004
  • An endoparasitoid wasp, Cotesia plutellae, has been used for a biological control agent against the diamondback moth, Plutellae xylostella. It has a symbiotic polydnavirus in their reproductive tract, which is required for its successful parasitization. Here, we measured a specific replication time of the polydnavirus during female development of C. plutellae. We, also, analyzed the reproductive potentials of female C. plutellae under mating or different host conditions. At $25^{\circ}C$, pupal C. plutellae began to develop adult tissues such as compound eyes and wings since day 2. At day 5, all adult tissues including antennae were developed and were ready to emerge. With polyclonal antibody raised against C. plutellae polydnavirus, an immunobloting could confirm virus replication at day 4 during pupal stage. Virus particles could be visualized by transmission electron microscope in the oviduct lumen of day 5 pupae. After adult eclosion, venom gland and ovarian calyx increased in size, though ovarioles did not. Mated females layed large number of eggs (over $60\%$) at first 4 days during their mean longevity of ca. 8 days at $25^{\circ}C$. Unmated females showed less active ovipositional behavior, where all the eggs developed into males. C. Plutellae parasitized both P. xylostella and fall webworm, Hyphantria cunea. However, C. Plutellae developed faster and showed higher successful paarasitization in P. xylostella than in H. cunea.

Risk Factors for Gallbladder Cancer in Nepal - a Case Control Study

  • Tamrakar, D;Paudel, IS;Adhikary, S;Rauniyar, B;Pokharel, PK
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.7
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    • pp.3447-3453
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    • 2016
  • Background: Gall bladder cancer (GBC) is highly fatal disease with poor prognosis, with a 5 year survival rate of <10%. It is relatively rare cancer worldwide; however it is the sixth cancer and second most common gastrointestinal tract cancer in Nepalese women. The study focused on associations of certain demographic, lifestyle, dietary, and reproductive factors with gall bladder cancer. Materials and Methods: We conducted a hospital-based matched case control study on newly diagnosed cases of primary GBC at BP Koirala Institute of Health Sciences and BP Koirala Memorial Cancer Hospital. Controls were healthy non-GBC relatives of cancer patients, matched for age, sex and marital status (in case of females) with cases at a ratio of 1:2. Data were collected between April 2012-April 2013 by semi structured interview from both cases and controls. Analyses were carried out with SPSS. Conditional logistic regression was used to find odds ratios and 95% confidence intervals for bivariate and multivariate analysis. Results: A total of 50 cases and 100 controls were enrolled in this study. On bivariate analysis, factors found to be significantly associated with gallbladder cancer were illiteracy (OR=3.29, CI=1.06-10.2), history of gallstone disease (OR=27.6, CI=6.57, 115.6), current smoker (OR=2.42, CI=1.005-5.86), early menarche <13 years (OR=2.64, CI=1.09-6.44), high parity more than 3 (OR=3.12, CI=1.25,7.72), and use of mustard oil (OR=3.63, CI=1.40, 9.40). A significant protective effect was seen with high consumption of fruits at least once a week (OR=0.101, CI=0.03-0.35). On multivariate analysis, history of gallstone disease, early menarche, current smoker and high consumption of fruits persisted as significant factors. Conclusions: History of gallstone disease, cigarette smoking and early menarche were associated with increased risk of gallbladder cancer while high consumption of fruits was found to have a protective effect.

Immunohistochemical study on the antigenicity of each organ structure of Clonorchis sinensis (간흡충 충체의 부위별 항원성에 대한 면역조직화학적 연구)

  • Jin Kim;Jong-Yil Chai;Weon-Gyu Kho;Kyu-Hyuk Cho;Soon-Hyung Lee
    • Parasites, Hosts and Diseases
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    • v.29 no.1
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    • pp.21-30
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    • 1991
  • An immunohistochemical study was performed to demonstrate comparative antigenicity of each body structure of the liver fluke, Clonorchis sinensis, such as the digestive tract, reproductive organs, excretory system, tegument and suckers. Indirect immunoperoxidase technique was applied, rising formalin-fixed and paraffin-embedded sections of C. sinensis as the antigen. Pooled cat sera obtained 10 weeks after an experimental infection with C. sinensis and peroxidase-conjugated goat anti-cat IgG were used as the primary and secondary antibodies, respectively. The intensity of immunohistochemical stain was very sensitive upon the titers of the primary and secondary antibodies, and their optimum dilutions were found to be 1:1,000∼1:2,000 and 1:1,000, respectively. The intestinal epithelial cells, intestinal content and excretory bladder showed strong positive coloring reactions even at lower titer (1 : 2,000) of the primary antibody, whereas the uterine wall and eggs, vitelline glands, and male reproductive organs showed only weak positive reactions despite an increase in the antibody titer (1:1,000). On the other hand, the suckers, tegument, subtegumental cells and other parenchyme portions did not reveal any positive immunoperoxidase reaction at the same antibody titers. From the above results, it is highly suggested that the most potent antigenicity of C. sinensis occur from their excretory-secretory substances originated from the digestive and excretory organs.

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The Effects of Vero Cell Co-culture on Mouse Embryo Development (Vero Cell과의 공동배양이 체외에서 생쥐 배아발생에 미치는 영향)

  • Lee, Yoon;Park, June-Hong;Kang, He-Na;Kim, Yong-Bong;Lee, Eung-Soo;Park, Sung-Kwan
    • Clinical and Experimental Reproductive Medicine
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    • v.24 no.2
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    • pp.233-239
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    • 1997
  • Embryos of most mammalian species grown in vitro would undergo developmental arrest at the approximate time of genomic activation. Stage-specific cell block and the resulting rapid loss of embryo viability in conventional culture media have limited the duration for which embryos may be cultured prior to transfer. As a result, embryos are usually transferred to the uterus at the 4-to 8-cell stage to avoid the loss of viability associated with long-term in vitro culture. Early transfer has led to asynchrony of the endometrium-trophectoderm interaction at the time of implantation and a resultant reduction in the rate of implantation. To overcome these problems, a variety of co-culture systems has been devised in which embryos can develop for a longer period prior to embryo transfer. Vero cells, derived from African green monkey kidney, share a common embryologic origin with cells from the genital tract. In addition, they are potentially safe to use, since they are highly controlled for viruses and other contaminants. Therefore, co-culture using Vero cells has been widely utilized to enhance embryo viability and development, although not without controversies. We thus designed a series of experiments to demonstrate whether Vero cells do indeed enhance mouse embryo development as well as to compare the efficacy of co-culturing mouse 1-cell embryos on Vero cell monolayer in both Ham's F-10 and human tubal fluid (HTF) culture media. 1-cell stage ICR mouse embryos were cultured either in the presence of Vero cells (Group A) or in conventional culture medium alone (Group B). In Ham's F-10 significantly more 3-to-8cell embryos developed in group A than group B (59.8 versus 10.0%; p<0.01). In contrast, there was no significant difference in embryonic development both group A and group B in HTF. However, significant differences were noted only in later embryonic stage (13 and 0%; p<0.05 of group A and B respectively, hatching or hatched). In Ham's F-10, we also could observe the beneficial effect of Vero cell on hatching process (70.7 and 42.1%; p<0.05 of group A and group B respectively).

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