• Title/Summary/Keyword: Human chorionic gonadotropin (hCG)

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Effect of hCG on Connexin 43 mRNA Expression in Goldfish Ovary

  • Choi, Cheol-Young
    • Journal of fish pathology
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    • v.16 no.3
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    • pp.215-217
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    • 2003
  • This study examined whether the connexin (Cx) is an essential protein during oocyte maturation in the ovary of the goldfish (Carassius auratus). In mature female goldfish ovaries, at late vitellogenic stage, human insulin-like growth factor-I (IGF-I; 20 M) and human chorionic gonadotropin(hCG; 20 IU/㎖) were injected. Twelve hr after the injection, mature female goldfish ovaries were removed and stored at -80C until analysis by RT-PCR. From the goldfish Cx43 cDNA sequence (GenBank accession number AB078505), two degenerate primers were designated. In vivo, 12 hr after the treatment with hCG, goldfish Cx43 mRNA expression level was increased, while the levels of IGF-I was not changed. Goldfish Cx43 mRNA expressed after, but not before the hCG treatment. These results suggest that Cx43 mRNA was judged to be a gene, which was transcribed during oocyte maturation induced by hCG.

Development and Immunochemical Properties of Two Monoclonal Antibodies Specific to Human Chorionic Gonadotropin

  • Kim, You-Hee;Koh, Kwan-Sam
    • BMB Reports
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    • v.32 no.5
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    • pp.474-479
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    • 1999
  • Using a hybridoma technique, spleen cells of Balb/c mice immunized with human chorionic gonadotropin (hCG) were fused with NS-1 mouse myeloma cells. Two hybrid cell lines, clones KS-8 and KS-19, secreting monoclonal antibodies to hCG, were isolated. KS-8 and KS-19 belong to the immunoglobulin $G_1$ subclass. With the aid of a double-antibody radioimmunoassay, it was established that the KS-8 monoclonal antibody recognizes an immunodeterminant of the $\beta$-subunit of hCG, whereas the KS-19 monoclonal antibody recognizes an epitope present on the $\alpha$-subunit of hCG. The KS-8 monoclonal antibody specifically reacts with human chorionic gonadotropin and shows cross-reactivity of less than 0.3% to other related human glycoprotein hormones. On the other hand, using a hemagglutination test based on antibody-induced agglutination of sheep red blood cells coated with hCG, It was shown that only the KS-19 monoclonal antibody was capable of inducing a positive reaction, although both monoclonal antibodies had similar binding capacity to the coated cells. The results from the dual screening procedures demonstrate that KS-8 and KS-19 monoclonal antibodies show high sensitivity in two different assays, and are hence useful for the qualitative and quantitative determination of hCG by both radioimmunoassay and hemagglutination inhibition tests.

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Development of recombinant human chorionic gonadotropin (hCG) using high-density culture technique of suspension-adapted chinese hamster ovary (CHO) cells

  • Na, Kyu-Heum;Kim, Seung-Chul;Seo, Kwang-Seok;Lee, Sung-Hee;Kang, Soo-Hyung
    • 한국생물공학회:학술대회논문집
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    • pp.37-37
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    • 2005
  • Human chorionic gonadotropin (hCG) is a heterodimeric glycoprotein hormone consisting of non-covalently linked two subunits, the ${\alpha}$ and ${\beta}$ subunit. It has been used as a infertility drug for ovulation to mimic luteinizing hormone $(LH).^{1)}$ A stable cell line was established by transfection of Rc/CMV-i-dhfr-hCG, expression vector containing hCG ${\alpha}-$ and ${\beta}-genes$, into dihydrofolate reductase-deficient CHO cells and subesquent methotrexate-mediated gene amplification. Anchorage-dependent CHO cells were adapted into a serum-free and/or animal component-free suspension medium through gradual serum weaning for the hCG production. The established cell line showed typical morphological characteristics and growth profile of CHO cells, and could produce FSH with passage-to-passage consistency. The high density perfusion culture of the CHO cells was carried out in Celligen Plus bioreactor equipped with a spin-filter as a internal cell retention device. The cell density reached up to $>1x10^{7}$ cells/ml in less than 7 days and a perfusion-control strategy based on cellular consumption rates of glucose was $established.^{2)}$ Biologically active recombinant hCG was purified by a series of chromatographic steps including anion exchange chromatography and hydrophobic interaction chromatography to homogeneity. The highly purified recombinant hCG was characterized for physicochemical, immunological and biological properties.

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Production of Monoclonal Antibody to Human Chorionic Gonadotropin(hCG) : Purification and Properties of a Monoclonal Antibody, and Immunochemiluminometric assay(ICMA) for the Assay of hCG (Human Chorionic Gonadotropin(hCG)에 대한 단일콜론항체 생산 : 단일클론항체의 분리정제 및 그 특성조사와 hCG정량을 위한 Immunochemiluminometric assay(ICMA)개발)

  • 최상훈;이병철;오재욱;이용환;서광영;정길생;김종배
    • Korean Journal of Animal Reproduction
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    • v.12 no.1
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    • pp.51-62
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    • 1988
  • Spleen cells of mouse immunized with hCG were fused with myeloma cell (SP 2/0 Ag 14) to produce monoclonal antibody against hCG. Several clones of hybridoma secreting monoclonal antibody were established and antibodies were characterized in terms of titer, subisotyping and sensitivity in immunoassay. Several methods, for the purification of anti¬bodies, based on gel-filtration, DEAE-ion exchange chromatography and affinity chromatography. were applied and compared each other by the result of SDS-PAGE. Two-site immunochemiluminometric assay (ICMA) involving the use of an excess concentration of a specific monoclonal antibody passively adsorbed onto the walls of plastic tubes and a chemiluminescence labelled antibody conjugate were de¬veloped for the determination of hCG as a preliminary study.

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Enzyme Immunoassay for Human Chorionic Gonadotropin Using Monoclonal Antibodies (단일크론성 항체를 이용한 융모막 성선자극 호르몬의 효소 면역측정법)

  • 차상훈;김희주;김원배;양중익
    • YAKHAK HOEJI
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    • v.31 no.2
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    • pp.64-69
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    • 1987
  • Monoclonal antibodies against human chorionic gonadotropin (hCG) were prepared and characterized by examining isotype, epitope binding, cross reactivity and affinity constants. And a sandwich type enzyme immunoassay for native hCG was developed with solid phase monoclonal antibody against the conformational determinant expressed only on native hCG and horseradish peroxidase conjugated monoclonal antibody against the $\beta$-subunit of hCG. The assay was sensitive to 1 mIU hCG/ml and shown a linear response up to 200 mIU hCG/ml. The cross reactivity for luteinizing hormone and $\beta$-subunit of hCG were 1% and 0.18%, respectively.

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Assay of Human Chorionic Gonadotropin in Urine of Athletes and Evaluation of Assay Kit Performance

  • Lee, Jung-Ran;Kim, Myung-Soo;Choi, Myung-Ja
    • Proceedings of the PSK Conference
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    • pp.406.2-406
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    • 2002
  • Special attention has been paid to human chorionic gonadotropin (hCG) for athlete doping control because it stimulates the endogenous production of testosterone and epitestosterone without increasing the T/E ratio which is a doping indicator for the exogenous administration of testosterone. Even though the IOC banned the use of hCG. a detection method has not been decided upon since there are a variety of immunoassay kits available on the market. We evaluated three kits in terms af their performance characteristics. (omitted)

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Nitric Oxide Generation from Peritoneal Macrophages by Human Chorionic Gonadotropin (사람 융모 성선 자극 호르몬에 의한 복강 대식세로로부터 산화질소의 발생)

  • Lee, Eun-Hee;Shin, Tae-Yong;Kim, Hyung-Min
    • YAKHAK HOEJI
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    • v.41 no.3
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    • pp.365-369
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    • 1997
  • Human chorionic gonadotropin (hCG) is a placental hormone and is involved in maintenance of the corpus luteum during pregnancy. In the present study, effect of hCG on nitiric ox ide (NO) generation from peritoneal macrophage was examined. hCG ahd no effect on NO generation by itself, whereas recombinant interferon- ${\gamma}$ (rIFN-${\gamma}$) alone had modest activity. When hCG was used in combination with rIFN-${\gamma}$, there was a marked cooperative induction of NO generation in a dose-dependent manner. The optimal effect of hCG on NO generation was shown at 6 hr after treatment with rIFN-${\gamma}$. Furthermore, northern blot analysis of showed that hCG increased the expression of inducible NO synthase(iNOS) gene. These results suggest that hCG induces NO generation from macrophages by increasing the expression of iNOS gene.

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Studies on the Effects of Follicular Environment and Human Chorionic Gonadotropin (hCG) on the Maturation of Rat Oocytes (흰쥐 난자의 성숙에 미치는 여포환경 및 hCG의 영향에 관한 연구)

  • Lee, Kyung-Ah;Rhee, Kun-Soo;Cho, Wan-Kyoo
    • The Korean Journal of Zoology
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    • v.28 no.4
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    • pp.245-256
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    • 1985
  • It has been found that the rat oocytes maintain germinal vesicle (GV) in general in the follicles either untreated or punctured, or in the foreign follicles for 17 hours culture unless they are cultured in the medium supplemented with human chorionic gonadotropin (hCG). That is, the proportion of oocytes with GV was in range of 88.8% and 95.2% in the plain medium, and on the other hand, only 11.1% to 19.4% of the oocytes were intact with GV when the follicles were exposed to hCG. The experiments with the oocytes which had once been cultured in the presence of dbcAMP or IBMX, and returned to the follicles for the additional culture showed almost the same results as above. That is, when the oocytes exposed to dbcAMP or IBMX for a certain length of period had been returned to the follicles, and set the additional culture, their maturation continuously suppressed even in the cultivation in the plain medium in which most of the oocytes usually resume meiosis. That is, despite of the cultivation in the plain medium, the oocytes transferred into the follicles failed to start maturation division, while the oocytes once exposed to the inhibitors immediately resume their maturation process in the inhibitor-free medium. Thus, it is apparent that the follicles provide inhibitory environment to the oocytes, and the inhibitory function is nullified by the presence of hCG.

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Estrus Behavior and Superovulatory Response in Black Bengal Goats (Capra hircus) Following Administration of Prostaglandin and Gonadotropins

  • Mishra, O.P.;Gawande, P.G.;Nema, R.K.;Tiwari, S.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.10
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    • pp.1374-1377
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    • 2004
  • The present study was conducted to explore the possibilities of estrus induction and superovulation in a native Indian breed of goats called 'Black Bengal'. Forty-two adult non-pregnant females were divided in two groups, of which 18 goats were subjected to a superovulatory treatment comprising of equine chorionic gonadotropin (eCG), Prostaglandin (PGF2$\alpha$) and human chorionic gonadotropin (hCG) to induce superovulation. The remaining 24 goats received no treatment and served as controls for the parameter under study as well as recipients for embryo transfer studies. The average duration of estrus was found to be significantly increased in treated goats (34.2${\pm}$3.4 h) compared to controls 3.0${\pm}$2.4 h). The average duration between PGF administration and occurrence of estrus was 2.0${\pm}$5.2 h. After mid ventral laparotomy, superovulatory responses indicated a significant increase in the number of follicles, which was 8.27${\pm}$0.37 in the treatment group compared to 4.16${\pm}$0.17 in the control group. The number of corpora lutea was also significantly increased in treated animals compared to control (2.90${\pm}$0.86 vs. 0.74${\pm}$0.04) respectively per ovary per goat.

Change of hCG Binding Capacity on the Granulosa Cell of Porcine Ovary During Follicular Atresia (돼지 난소내 여포의 폐쇄과정중 과립세포의 결합능의 변화)

  • Chang, Choul-Soo;Lee, Chang-Joo;Yoon, Yong-DaI;Kim, Moon-Kyoo
    • Clinical and Experimental Reproductive Medicine
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    • v.13 no.1
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    • pp.29-38
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    • 1986
  • In order to study the mechanism of follicular atresia, follicles were classified into the normal groups and the atretic ones, according to the criteria with or without corpus luteum, size of follicles, vascularization, status of granulose cells and the hypertrophy of theca layers in the porcine ovary. To estimate the binding capacity of human chorionic gonadotropin (hCG) receptor on the granulosa cells during atresia, hCG were iodinated by the chloramine-T method and then purified through the column chromatography. The concentration" of hCG receptor in each group were measured by the hCG receptor binding assay. Binding capacity in large normal follicles were 1.16%, but atretic ones were 0.45%. But in medium and small follicles (below 6mm in diameter), the binding capacity in normal follicles were 0.09%, but atretic ones were 0.05%, which was lower than those of large follicles. The present ( ) that the concentrations of hCG receptors on granulosa cells is decreased when the follicles become atretic and be used as a sort of creteria for the identification of follicles atresia.

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