• Title/Summary/Keyword: C-ELISA

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Immunogenicity of 7-valent pneumococcal conjugate vaccine related to booster immunization in Korean children (한국 소아에서 7가 폐렴사슬알균 단백결합 백신의 추가접종 면역원성에 관한 연구)

  • Park, So Eun;Lee, Hyunju;Lim, Soo Young;Kim, Kyung Hyo
    • Clinical and Experimental Pediatrics
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    • v.51 no.6
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    • pp.622-628
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    • 2008
  • Purpose : The purpose of this study was to evaluate the immunogenicity of the booster immunization with pneumococcal conjugate vaccine in Korean children. Methods : Thirty-nine children aged 12-23 months who visited Kangnam CHA Hospital between September 2006 and December 2006 were enrolled. The children were divided into primary and booster groups depending on their vaccination status for the 7-valent pneumococcal conjugate vaccine. The anti-pneumococcal antibody levels of each serotype included in the vaccine (4, 6B, 9V, 14, 18C, 19F, 23F) were determined by third-generation ELISA. Results : The geometric mean titer (GMT) of antibodies to each pneumococcal serotype in the booster group was higher than in the primary group (P<.05). The percentage of subjects with pneumococcal antibodies ${\geq}0.35{\mu}g/mL$ was 90.5-100% for all serotypes in both the primary and booster groups. The percentage of subjects with pneumococcal antibodies ${\geq}1.0g/mL$ in the booster group was 94.4-100%, which was higher than the primary group except for serotypes 6B and 14 (P<.05). The percentage of subjects with pneumococcal antibodies ${\geq}5.0{\mu}g/mL$ in the booster group was 50.0-94.4% which was higher than the primary group for all serotypes (P<0.05). Conclusion : The immunogenicity of a booster dose of the pneumococcal conjugate vaccine in Korean children was high and the immunogenicity of a primary series was also relatively high. To determine the feasibility of the introduction of the pneumococcal conjugate vaccine and the appropriate schedule for Korean children, further prospective investigation of the immunogenicity of the booster immunization is needed.

Adjuvant Effect of PAMAM Dendrimer on the Antigenicity of Keyhole Limpet Hemocyanin in Balb/c Mice (Balb/c 마우스에서 Keyhole limpet hemocyanine (KLH)의 항원성에 대한 PAMAM dendrimer 의 면역증강 효과)

  • Lee, Ga-Young;Kim, Min Jee;Kim, So Yeon;Lee, Kyung Bok;Oh, Dong Hyun;Cho, Young Ho;Yoo, Yung Choon
    • Journal of Life Science
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    • v.30 no.10
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    • pp.905-911
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    • 2020
  • The adjuvant effect of PAMAM dendrimer G4 (PAMAM) on the induction of humoral and cellular immune responses against keyhole limpet hemocyanin (KLH) was examined. Mice were immunized subcutaneously twice at two-week intervals with KLH, with or without PAMAM dendrimer (100 ㎍/mouse), and the mice immunized with KLH+PAMAM showed significantly higher antibody titers against KLH than those immunized with KLH alone. The assay for determining the isotypes of the antibodies showed that PAMAM augmented the KLH-specific antibody titers of IgG1, IgG2a, IgG2b, IgG3, and IgM. In addition, mice immunized twice with KLH+PAMAM followed by a subcutaneous injection of KLH (20 ㎍/site) 7 weeks after the primary immunization exhibited a higher delayed-type hypersensitivity (DTH) reaction than those treated with KLH alone. In an in vitro analysis of T lymphocyte proliferation in response to KLH in week 8, the splenocytes of mice treated with KLH+PAMAM showed significantly higher proliferating activity than those treated with KLH alone, and the culture supernatants of cell cultures from mice immunized with added PAMAM dendrimer showed higher levels of KLH-specific cytokine (IL-4 and IFN-r) production. These results suggest that PAMAM dendrimer G4 possesses a potent immune-adjuvant activity for enhancing both humoral and cell-mediated immunity specific to foreign antigens.

Antibody Responses in Hematopoietic Cell Transplantation Recipients after Vaccination Against Haemophilus Influenzae Type b and Streptococcus pneumoniae (소아 조혈모세포 이식 환자에서 b형 헤모필루스 인플루엔자와 폐렴구균 백신 접종 후 항체 반응에 관한 연구)

  • Kim, Yae-Jean;Hwang, Ji-Young;Choi, Soo-Han;Kong, Eunhye;Kim, Yanghyun;Park, Ki-Sup;Yoo, Keon-Hee;Sung, Ki-Woong;Koo, Hong Hoe;Kim, Kyung-Hyo
    • Pediatric Infection and Vaccine
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    • v.21 no.2
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    • pp.81-95
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    • 2014
  • Purpose: Hematopoietic cell transplantation (HCT) recipients are vulnerable to invasive infection by Haemophilus influenzae type b (Hib) and Streptococcus pneumoniae (Sp). This study was performed to evaluate immune responses after Hib and Sp vaccination in Korean pediatric HCT recipients. Methods: Patients were prospectively enrolled at Samsung Medical Center during 2009-2011. ELISA tests to detect anti-PRP IgG antibody and antibodies to Sp serotypes 4, 6B, 9V, 14, 18C, 19F, and 23F were performed at the Center for Vaccine Evaluation and Study, Ewha Medical Research Institute. Results: Ten patients (two allogeneic, eight autologous recipients) with median age 5.4 years (range 2.7-12.2 years) were enrolled. Before Hib vaccination, 60% of patients' anti-PRP IgG titers were below $0.15{\mu}g/mL$. After vaccination, 100% of patients' anti-PRP IgG titers increased above $0.15{\mu}g/mL$ (cut-off value for detection) and $1.0{\mu}g/mL$ (cut-off value for seroprotection). For pneumococcus, in 2-5 year-old patients, pre-vaccination geometric mean concentrations (GMCs) of IgG for six serotypes (4, 6B, 9V, 14, 18C, and 23F) were below $0.35{\mu}g/mL$ and at 5 months post-vaccination GMCs of IgG for all seven serotypes increased to above $0.35{\mu}g/mL$. In patients older than 5 years, pre-vaccination GMCs of IgG for four serotypes (4, 9V, 14, and 23F) were below $0.35{\mu}g/mL$ and at 3 months post-vaccination GMCs of IgG for all seven serotypes increased to above $0.35{\mu}g/mL$. Conclusion: Most HCT recipients had low or no protective antibodies to Hib and Sp before vaccination, but showed good immune responses to protective levels after vaccination.

Gene Expression of Metalloproteinases, Tissue Inhibitors of Metalloproteinases and Cytokines in Adriamycin-induced Cardiomyopathy (아드리아마이신으로 유도된 심근증에서 Metalloproteinase, Metalloproteinase 조직억제자, Cytokine 유전자 발현에 대한 연구)

  • Hong, Young Mi
    • Clinical and Experimental Pediatrics
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    • v.48 no.2
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    • pp.197-203
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    • 2005
  • Purpose : Changes in metalloproteinases(MMP) activity have been demonstrated in several disease states, including rheumatoid arthritis and tumor metastasis. More importantly, increased myocardial MMP activity has been reported to occur in both clinical and experimental forms of dilated cardiomyopathy. There was no report about MMP in adriamycin(ADR)-induced cardiomyopathy. The purpose of this study was to investigate gene expression of MMP and tissue inhibitor of metalloproteinases(TIMP) in ADR-induced cardiomyopathy and clarify the relationship between MMP and cytokines. Methods : Male Sprague-Dawley rats were divided into two groups. The first group was control. The second group was given intraperitoneal injections of ADR(5 mg/kg) twice a week over two weeks. Serum concentrations of MMP, TIMP, interleukin(IL)-6 and tumor necrosis factor(TNF)-${\alpha}$ were measured. RNA extraction was performed from frozen rat hearts. Reverse transcription polymerase chain reaction(RT-PCR) was employed. cDNA Microarray analysis was performed by using a set of 5,184 sequence-verified rat cDNA clones. Results : Serum MMP and TIMP levels were not significantly different between the two groups. IL-6 was $36.8{\pm}2.8pg/mL$ and TNF-${\alpha}$ $2.2{\pm}2.7pg/mL$ in the ADR group. They were significantly higher than in the control group. Serum MMP correlated significantly with TNF-${\alpha}$(r=0.41, P<0.05). There was no gene expression of MMP, IL-6 or TNF-${\alpha}$ in the hearts of both groups. Gene expression of TIMP was significantly depressed in the hearts of the ADR group. Conclusion : These results suggested a potential role for TNF-${\alpha}$ in the regulation of extracellular matrix remodeling in ADR induced cardiomyopathy. Rapid screening of multiple decreased gene expression by DNA chip may be a useful diagnostic test to detect early cardiac injury before developing ADR induced cardiomyopathy.

Mechanism by which periodontitis may contribute to atherosclerosis (치주염이 동맥경화에 기여하는 기전에 관한 연구)

  • Han, Seung-Hee;Kim, Kyung-Hwa;Yang, Seung-Min;Chung, Hyun-Ju;Choi, Yoon-Sik;Han, Soo-Boo;Chung, Chong-Pyoung;Rhyu, In-Chul
    • Journal of Periodontal and Implant Science
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    • v.32 no.4
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    • pp.837-846
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    • 2002
  • 그람 음성균의 감염에 의한 만성 염증질환인 치주염이 동맥경화를 동반한 허혈성 심장질환 (협심증이나 심근 경색)을 일으킬 수 있는 위험인자로 작용할 수 있다는 보고가 있었다. 그러나, 그 기전에 관해서는 명확하게 알려져 있지 않다. 작용기전의 하나로서, 치주염에 의해 치주조직에서 국소적으로 생긴 염증성 싸이토카인(IL-1${\beta}$, IL-6, $PGE_2$, TNF-${\alpha}$)이 혈행을 따라 이동하여 심혈관에서 동맥경화를 일으킬 수 있다는 가설이 제시되고 있는데, 이 가설을 검증해 보고자 한다. 서울대학교 병원 순환기 내과에 불안정 협심증이나 심근경색으로 입원한 환자 및 과거 이 질환의 병력을 갖고 있거나 검진 목적으로 내원하여 관상동맥 조형술을 받은 환자들 중 동맥경화로 진단받은 사람을 실험군(24명)으로 하고, 동맥경화로 진단받지 않은 사람을 대조군(12명)으로 하였다. 치주질환의 활성도를 나타내는 치은 지수, 치태 지수, 치주낭 깊이, 부착 상실을 측정하였다. Paper strip을 실험대상 치아(Ramfjord's teeth)들 중에서 가장 깊은 치주낭을 가진 두 개의 치아를 택하여 각 치아의 가장 깊은 치주낭에 30초간 삽입한 후 밀폐된 plastic tube에 넣고 ELISA kit를 이용하여 IL-1${\beta}$, IL-6, TNF-${\alpha}$, $PGE_2$의 농도를 측정하였다. 환자의 plasma에서도 동일한 싸이토카인의 농도를 측정하였다. 설문조사를 통해 동맥경화의 위험 인자로 간주되어온 고혈압, 당뇨, 가족력, 심근경색이나 협심증의 기왕력, 흡연의 유무를 기록하였다. 혈액검사를 하여 total cholesterol, triglycerides, LDL cholesterol, HDL cholesterol, WBC, CRP (C-reactive protein)의 농도를 측정하였다. 치주조직에 대한 임상 검사 결과 치은의 염증상태를 나타나는 지표인 치은 지수에서만 실험군이 대조군에 비해 유의할 정도 (p=0.0174)로 높게 나타났고, 만성적인 염증의 결과로 인한 치조골의 사실 정도를 나타내는 치주낭 깊이나 부착 상실에서는 유의할 만한 차이를 보이지 않았다. 치주낭에서 측정한 염증성 싸이토카인 중 IL-1${\beta}$, $PGE_2$가 실험군에서 유의할 만한 차이 (p=0.005, 0.022)를 보이며, 더 높은 농도로 나타났고, TNF-${\alpha}$는 대조군에서 유의성 있게 (p=0.009) 높게 나타났다. 그러나, plasma의 싸이토카인이나, serum lipid/lipoprotein, C-reactive Protein, WBC는 유의할 만한 차이를 보이지 않았다. 또한 치은열구액내의 싸이토카인과 이에 상흥하는 싸이토카인 간에 상관관계는 관찰되지 않았다. 다변량 로지스틱 회귀분석 결과, 치은열구액내의 IL-1${\beta}$와 TNF-${\alpha}$ 만이 동맥경화와 유의성 있는 관련성을 보였고, 특히 IL-1${\beta}$와 교차비는 273으로 상당한 관련성을 보여주었다. 결론적으로, 치주조직에서 국소적으로 생긴 염증성 싸이토카인이 그대로 혈행으로 이동하여 혈장내의 싸이토카인 농도를 높이는 것은 아니다. 그러나, 치주염으로 인해 치은열구내액내에 국소적으로 증가된 염증성 싸이토카인은 동맥경화와 상당한 관련성을 가진다.

Anti-inflammatory effect of Sinhyowoldo-san Extract with regard to Pro-inflammatory Mediators in PMA plus A23187-induced Human Mast Cells (인간 비만세포에서 PMA와 A23187에 의해 유도된 전염증 매개체에 대한 신효월도산 추출물의 항염증 효과)

  • Wi, Gyeong;Yang, Da-Wun;Kang, Ok-Hwa;Kim, Sung-Bae;Mun, Su-Hyun;Seo, Yun-Soo;Kang, Da-Hye;Lim, Jae-Soo;Kim, Ma-Ryong;Kwak, Nam-Won;Kong, Ryong;Kwon, Dong-Yeul
    • The Korea Journal of Herbology
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    • v.29 no.6
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    • pp.117-123
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    • 2014
  • Objectives : Sinhyowoldo-san (SHWDS) is said to be a traditional medicine used for shigellosis, abdominal pain, diarrhea. But mechanism of SHWDS mediated-modulation of immune function is not sufficiently understood. To ascertain the molecular mechanisms of SHWDS 70% EtOH extract on pharmacological and biochemical actions in inflammation, we researched the effect of pro-inflammatory mediators in phorbol-12-myristate-13-acetate (PMA)+ A23187-activated human mast cell line (HMC-1). Methods : In the present research, cell viability was measured by MTS assay. pro-inflammatory cytokine production was measured by performing enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase chain reaction (RT-PCR), and western blot analysis to analyze the activation of mitogen-activated protein kinases (MAPKs), nuclear factor kappa-light-chain-enhancer of activated B cells ($NF-{\kappa}B$). The investigation focused on whether SHWDS inhibited the expressions of interleukin-6 (IL-6), interleukin-8 (IL-8), MAPKs and $NF-{\kappa}B$ in PMA+A23187-activated HMC-1 cells. Results : SHWDS has no cytotoxicity at measured concentration (50, 100, and $250{\mu}g/ml$). SHWDS ($250{\mu}g/ml$) inhibits pro-inflammatory cytokine expression in PMA+ A23187-activated HMC-1 cells. Moreover, SHWDS inhibited cyclooxygenase (COX)-2 expression. In activated HMC-1 cells, SHWDS suppressed phosphorylation of extracellular signal-regulated kinase (ERK 1/2) and c-jun N-terminal Kinase (JNK 1/2). Then, SHWDS suppressed activation of nuclear factor $NF-{\kappa}B$ in nuclear, degradation of IkB ${\alpha}$ in cytoplasm. Conclusions : We propose that SHWDS has an anti-inflammatory therapeutic potential, which may result from inhibition of ERK 1/2, JNK 1/2 phosphorylation and $NF-{\kappa}B$ activation, thereby decreasing the expression of pro-inflammatory genes.

A Co-inhibitory Molecule, B7-H4, Synergistically Potentiates Oral Tolerance by Inducing CD4+CD25+FoxP3+ T Cells

  • Wen, Lanying;Yang, Sung-Yeun;Choi, Jae-Kyoung;Kim, Young-Hee;Kwon, Eun-Hee;Lee, Hyun-Ji;Jeoung, Hae-Young;Hwang, Du-Hyeon;Hwang, Dong-Jin;Choi, In-Hak
    • IMMUNE NETWORK
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    • v.8 no.1
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    • pp.21-28
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    • 2008
  • Background: A co-inhibitory molecule, B7-H4, is believed to negatively regulate T cell immunity by suppressing T cell proliferation and inhibiting cytokine production. However, the mechanism behind B7-H4-mediated tolerance remains unclear. Methods: Balb/c $(H-2^d)$ mice were fed with dendritic cell line, DC2.4 $(H-2^d)$ every day for 10 days. Meantime, mice were hydrodynamically injected with recombinant plasmid expressing B7-H4 fusion protein (B7-H4.hFc) or hFc via tail vein. One day after last feeding, mice were immunized with allogeneic B6 spleen cells. 14 days following immunization, mice were challenged with B6 spleen cells to ear back and the ear swelling was determined the next day. Subsequently, a mixed lymphocyte reaction (MLR) was also performed and cytokines profiles from the reaction were examined by sandwich ELISA. Frequency of immunosuppressive cell population was assayed with flow cytometry and mRNA for FoxP3 was determined by RT-PCR. Results: Tolerant mice given plasmid expressing B7-H4.hFc showed a significant reduction in ear swelling compared to control mice. In addition, T cells from mice given B7-H4.hFc plasmid revealed a significant hyporesponsiveness of T cells against allogeneic spleen cells and showed a significant decrease in Th1 and Th2 cytokines such as IFN-${\gamma}$, IL-5, and TNF-${\alpha}$. Interestingly, flow cytometric analysis showed that the frequency of CD4+CD25+FoxP3+ Tregs in spleen was increased in tolerant mice given recombinant B7-H4.hFc plasmid compared to control group. Conclusion: Our results demonstrate that B7-H4 synergistically potentiates oral tolerance induced by allogeneic cells by increasing the frequency of FoxP3+ CD4+CD25+ Treg and reducing Th1 and Th2 cytokine production.

Response of Measles-specific Antibody in Children with Measles During Measles Epidemic in Seongnam, 1993 (1993년도 성남지역에서 유행한 홍역 환아에서의 홍역 특이 항체 반응)

  • Kim, Ji Youn;Park, Yoon Hyung;Kim, Soon Ki;Choi, Yun Hwa;Lee, Hoan Jong;Son, Byong Kwan
    • Pediatric Infection and Vaccine
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    • v.4 no.1
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    • pp.126-132
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    • 1997
  • Measles outbreak in the world was decreased since measles vaccine had been introduced. Although vaccination rate is high, measles was not eradicated and measles reappeared among vaccinated children. We measured measles-specific antibody from the vaccinated and unvaccinated groups who had experienced apparent measles in the Seongnam city in 1993. The results were as follows. 1) The data included total 126 children (M:F=1 : 1). Age distribution of measles outbreak revealed 6 children in 5yr, 11 in 6yr, 20 in 7yr, 39 in 8yr, 22 in 9yr, 11 in 10yr, 11 in 11yr, and 6 in 12yr. 2) MMR vaccination rate was 78.6%(99/126) in the children who had experienced measles. Positive rate of measles-specific IgM Ab was 80.8% (80/99) among the vaccinated group and among 9E.6.% (25/27) the unvaccinated. 3) Positive rate of measles-specific IgG Ab was 90.9% (90/99) among MMR-vaccinated group, and 85.2% (23/27) in unvaccinated group. In conclusion, measles-specific IgM antibody have been detected more than 1 month in most patients. The relatively high proportion of measles-specific IgM positivity may mean primary vaccine failure. To booster the antibody titers and to prevent measles epidemic in school-aged children, revaccination of measles should be considered.

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TNF-α stimulated IL-8 and IL-10 expression in monocytes from patients with chronic granulomatous disease (만성육아종질환 환자 단핵구에서 TNF-α 자극에 의한 IL-8과 IL-10의 발현 양상)

  • Shin, Kyung-Sue
    • Clinical and Experimental Pediatrics
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    • v.51 no.10
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    • pp.1096-1101
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    • 2008
  • Purpose : Patients with chronic granulomatous disease (CGD) have genetic mutations in a component of the NADPH oxidase enzyme that is necessary for the generation of the superoxide anion. The profound defect in innate immunity is reflected by the patients susceptibility to catalase-positive bacteria and fungi. In addition, CGD patients display signs of persistent inflammation, which is not associated only with deficient superoxide anion production. The aim of this study was to elucidate the cytokine responses in CGD patients after $TNF-{\alpha}$ stimulation. Methods : Heparinized blood samples were collected from 8 CGD patients and 10 healthy volunteers. Monocytes ($1{\times}10^6cell/well$) isolated by the magnet cell isolation system were incubated with a constant amount of $TNF-{\alpha}$ (10 ng/mL) at $37^{\circ}C$ for 6 h. Incubated cells were harvested at 60-min intervals for IL-8 and IL-10 mRNA analysis, and the supernatant was collected at the same intervals to determine IL-8 and IL-10 expression. Monocytes from healthy volunteers were also incubated with antioxidants followed by $TNF-{\alpha}$ stimulation for IL-8 and IL-10 expression. Results : In CGD patients, a high expression of IL-8 together with a significantly higher IL-10 expression than in the healthy controls was seen after $TNF-{\alpha}$ stimulation. Moreover, normal monocytes treated with antioxidants exhibited increased IL-8 responses. Conclusion : The absence of phagocyte-derived reactive oxidants in CGD might be associated with a dysregulated production of pro- and antiinflammatory cytokines. Additional research related to reactive oxidants is needed to clarify the role of cytokines in CGD patients.

Administration of Anti-Inhibin Serum and Bovine Follicular Fluid on Plasma Hormonal Level and Ovarian Follicular Development in Hanwoo (Anti-Inhibin Serum 및 Bovine Follicular Fluld 처리가 한우 난소의 난포 발달과 혈중 호르몬 변화에 미치는 영향)

  • Seong, H.H.;Yang, B.C.;Hwang, S.S.;Park, S.J.;Ko, Y.G.;Rho, W.K.;Yang, B.S.
    • Journal of Embryo Transfer
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    • v.21 no.4
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    • pp.315-322
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    • 2006
  • This study was performed to investigate the physiological effects of bovine follicular fluid (bFF) or anti-inhibin serum (AI) on follicular development in Hanwoo. Saline (0.95%), bFF or AI (total of 40 ml) were administered into the jugular vein in 9 Hanwoo cows. The plasma inhibin, estradiol-17 $\beta$ (E2), and progesterone (P4) levels were measured using RIA or ELISA kit and the number of ovarian follicles was observed by ultrasonography at 72 hr after ovulation. The plasma inhibin level in bFF treatment group was significantly increased and maintained higher level from 102 hr after ovulation compared to that of saline and AI groups (p<0.05). In plasma E2 level, AI treatment group showed significantly higher level from 36 hr to 108 hr after ovulation than that of saline and bFF groups (p<0.05). After that it showed decreasing tendency. The plasma P4 level was increased in control and AI treatment groups at 68 hr after ovulation. However, it was maintained significantly lower level in bFF group from 84 hr to 180 hr compared to that of saline and AI group (p<0.05). As a result of ultrasonography at 72 hr after ovulation, higher number of follicles was shown in AI treatment group compared to bFF groups, although the difference was not statistically significant. Taken together, it can be postulated that a treatment of synthesized AI inhibits the secretion of inhibin, stimulates FSH secretion inhibited by inhibin, and induces follicular development and estrogen secretion. According to these results, a development of ovarian follicle immediately after ovulation is associated closely with inhibin in Hanwoo heifers.