Yam Kwan Mei. / Thesis submitted in: August 1999. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references (leaves 108-129). / Abstracts in English and Chinese. / Abstract (in English) --- p.ii / Abstract (in Chinese) --- p.iv / Acknowledgement --- p.vi / Table of Contents --- p.ix / List of Tables --- p.xiv / List of Figures --- p.xv / Symbols and Abbreviations --- p.xviii / Scientific names --- p.xx / Chapter Chapter 1 --- General Introduction --- p.1 / Chapter 1.1 --- Gonadotropins --- p.1 / Chapter 1.1.1 --- Structure --- p.1 / Chapter 1.1.2 --- Function --- p.3 / Chapter 1.1.3 --- Regulation --- p.9 / Chapter 1.1.3.1 --- GnRH --- p.9 / Chapter 1.1.3.2 --- Steroids --- p.11 / Chapter 1.1.3.3 --- Activin --- p.12 / Chapter 1.2 --- Activin Family of Growth Factors --- p.14 / Chapter 1.2.1 --- Structure --- p.14 / Chapter 1.2.2 --- Function --- p.17 / Chapter 1.3 --- Objectives --- p.22 / Chapter Chapter 2 --- Cloning of Goldfish Activin βA cDNA and the Expression of Its mRNA in Gonadal and Non-gonadal Tissues --- p.24 / Chapter 2.1 --- Introduction --- p.24 / Chapter 2.2 --- Materials and Methods --- p.25 / Chapter 2.2.1 --- Cloning of goldfish activin βA cDNA --- p.26 / Chapter 2.2.1.1 --- Cloning of the 5' and 3' cDNA ends --- p.26 / Chapter 2.2.1.2 --- Extension of the 5' and 3' fragments --- p.28 / Chapter 2.2.2 --- Sequencing of the cDNA --- p.28 / Chapter 2.2.2.1 --- Generation of pKS/GactβA constructs with insert in different orientations --- p.28 / Chapter 2.2.2.2 --- Generation of overlapping subclones --- p.29 / Chapter 2.2.2.3 --- Cycle sequencing --- p.30 / Chapter 2.2.2.4 --- Sequence analyses --- p.30 / Chapter 2.2.3 --- Isolation of total and messenger RNA --- p.30 / Chapter 2.2.3.1 --- Isolation of total RNA --- p.30 / Chapter 2.2.3.2 --- Isolation of messenger RNA --- p.31 / Chapter 2.2.4 --- Southern blot analysis --- p.32 / Chapter 2.2.5 --- Northern blot analysis --- p.33 / Chapter 2.2.6 --- Reverse transcription-polymerase chain reaction (RT-PCR) --- p.33 / Chapter 2.3 --- Results --- p.35 / Chapter 2.3.1 --- Cloning and sequence analysis of activin β A cDNA --- p.35 / Chapter 2.3.2 --- Distribution of activin βA mRNA in different tissues --- p.49 / Chapter 2.4 --- Discussion --- p.53 / Chapter Chapter 3 --- Establishment and Characterization of Stable Cell Lines for the Recombinant Production of Goldfish Activin A --- p.59 / Chapter 3.1 --- Introduction --- p.59 / Chapter 3.2 --- Materials and Methods --- p.60 / Chapter 3.2.1 --- Construction of expression plasmid --- p.60 / Chapter 3.2.2 --- Cell culture --- p.62 / Chapter 3.2.3 --- Transfection of CHO cells --- p.62 / Chapter 3.2.4 --- G418 selection of transfected CHO cells --- p.62 / Chapter 3.2.5 --- Activin bioassay (EDF-assay) --- p.63 / Chapter 3.2.6 --- Cloning of pBK/GactβA-transfected CHO cells by limited dilution --- p.63 / Chapter 3.2.7 --- Isolation of total RNA --- p.65 / Chapter 3.2.8 --- Northern blot analysis --- p.65 / Chapter 3.3 --- Results --- p.66 / Chapter 3.3.1 --- Optimization of G418 concentration for selection --- p.66 / Chapter 3.3.2 --- Expression of activin activity by pBK/GactpβA- transfected CHO cells --- p.67 / Chapter 3.3.3 --- Establishment and characterization of CHO cell lines that stably produce recombinant goldfish activin A --- p.67 / Chapter 3.4 --- Discussion --- p.73 / Chapter Chapter 4 --- Differential Regulation of Goldfish Gonadotropin (GTH-Iβ and GTH-IIβ) Gene Expression by Recombinant Goldfish Activin --- p.79 / Chapter 4.1 --- Introduction --- p.79 / Chapter 4.2 --- Materials and Methods --- p.82 / Chapter 4.2.1 --- Animals --- p.82 / Chapter 4.2.2 --- Drug treatment --- p.83 / Chapter 4.2.3 --- Primary culture of dispersed pituitary cells --- p.84 / Chapter 4.2.4 --- Southern blot analysis --- p.85 / Chapter 4.2.5 --- Isolation of total RNA --- p.86 / Chapter 4.2.6 --- Northern blot analysis --- p.86 / Chapter 4.2.7 --- Dot blot analysis --- p.87 / Chapter 4.2.8 --- Data analyses --- p.87 / Chapter 4.3 --- Results --- p.88 / Chapter 4.3.1 --- Probe specificity --- p.88 / Chapter 4.3.2 --- Effects of goldfish activin on pituitary GTH-Iβ and -IIβ mRNA expression --- p.88 / Chapter 4.3.3 --- Blockade of activin effects by follistatin --- p.92 / Chapter 4.4 --- Discussion --- p.96 / Chapter Chapter 5 --- General Discussion --- p.101 / Chapter 5.1 --- Overview --- p.101 / Chapter 5.2 --- Contribution of the Present Research --- p.103 / Chapter 5.2.1 --- Cloning of full-length goldfish activin βA cDNA --- p.103 / Chapter 5.2.2 --- Establishment of stable cell lines for the recombinant production of goldfish activin A --- p.104 / Chapter 5.2.3 --- Differential regulation of goldfish gonadotropin (GTH-Iβ and GTH-IIβ) gene expression by recombinant goldfish activin --- p.105 / Chapter 5.3 --- Future Research Direction --- p.107 / Chapter 5.3.1 --- Activin studies --- p.107 / Chapter 5.3.2 --- Gonadotropin studies --- p.107 / References --- p.108

Poon, Shui-Kei. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 78-98). / Abstracts in English and Chinese. / Abstract (in English) --- p.ii / Abstract (in Chinese) --- p.iv / Acknowledgement --- p.vi / Table of content --- p.vii / List of figures --- p.x / Symbols and abbreviations --- p.xii / Chapter 1 General Introduction / Chapter 1.1 --- Structure of ovarian follicles --- p.1 / Chapter 1.2 --- Folliculogenesis and its control --- p.1 / Chapter 1.2.1 --- History of inhibin discovery --- p.4 / Chapter 1.2.2 --- Inhibin and its related proteins --- p.5 / Chapter 1.3 --- Inhibin --- p.6 / Chapter 1.3.1 --- Structure --- p.6 / Chapter 1.3.2 --- Function --- p.7 / Chapter 1.3.3 --- Signaling --- p.11 / Chapter 1.3.4 --- Expression --- p.14 / Chapter 1.3.5 --- Regulation --- p.15 / Chapter 1.4 --- Objectives of the present study --- p.19 / Chapter Chapter 2 --- Spatiotemperoal Expression Profiles of Inhibin a in the Zebrafish Ovary / Chapter 2.1 --- Introduction --- p.25 / Chapter 2.2 --- Materials and Methods --- p.27 / Chapter 2.2.1 --- Animals --- p.27 / Chapter 2.2.2 --- Chemicals --- p.27 / Chapter 2.2.3 --- Separation of oocytes and follicular layers --- p.28 / Chapter 2.2.4 --- Isolation of ovarian follicles --- p.28 / Chapter 2.2.5 --- RNA isolation and reverse transcription --- p.28 / Chapter 2.2.6 --- Real-time and semi-quantitative RT-PCR quantification of expression --- p.29 / Chapter 2.2.4 --- Data analysis --- p.30 / Chapter 2.3 --- Results --- p.30 / Chapter 2.3.1 --- Validation of semi-quantitative RT-PCR quantification --- p.30 / Chapter 2.3.2 --- Tissue distribution of inha expression --- p.30 / Chapter 2.3.3 --- Localization of inha expression within the ovarian follicle --- p.31 / Chapter 2.3.4 --- Further evidence for inha expression in the follicle layer --- p.31 / Chapter 2.3.5 --- Stage-dependent expression of inha in the ovarian follicles --- p.32 / Chapter 2.4 --- Discussion --- p.33 / Chapter Chapter 3 --- Regulation of Inhibin a Expression in vitro and in vivo and the Effect of Inhibin on Final Oocyte Maturation / Chapter 3.1 --- Introduction --- p.43 / Chapter 3.2 --- Materials and Methods --- p.46 / Chapter 3.2.1 --- Animals --- p.46 / Chapter 3.2.2 --- Chemicals and hormones --- p.46 / Chapter 3.2.3 --- Preparation of goldfish pituitary extract --- p.47 / Chapter 3.2.4 --- Ovarian fragment incubation --- p.47 / Chapter 3.2.5 --- Preparation of spontaneously matured follicles --- p.48 / Chapter 3.2.6 --- Intra-peritoneal injection --- p.48 / Chapter 3.2.7 --- RNA isolation and reverse transcription --- p.48 / Chapter 3.2.8 --- Real-time and semi-quantitative RT-PCR quantification of expression --- p.48 / Chapter 3.2.9 --- Data analysis --- p.49 / Chapter 3.3 --- Results --- p.49 / Chapter 3.3.1 --- Validation of semi-quantitative RT-PCR quantification --- p.49 / Chapter 3.3.2 --- Temporal change of basal inha expression in cultured ovarian fragments --- p.49 / Chapter 3.3.3 --- Effect of pituitary extract on the expression of inha --- p.50 / Chapter 3.3.4 --- Effects of recombinant zebrafish FSH and LH on inha expression --- p.51 / Chapter 3.3.5 --- Effect of forskolin on inha expression --- p.52 / Chapter 3.3.6 --- Involvement of PKA and p38MAPK in the pituitary extract and forskolin-induced up-regulation of inha expression --- p.52 / Chapter 3.3.7 --- Effect of recombinant zfFSH on the expression of inha in vivo --- p.53 / Chapter 3.3.8 --- Effects of inhibin on the basal and DHP-induced final oocyte maturation --- p.53 / Chapter 3.4 --- Discussion --- p.54 / Chapter Chapter 4 --- General Discussion --- p.71 / References --- p.78

by Wong, Cheuk-fai. / Thesis (M.Sc.)--Chinese University of Hong Kong, 1994. / Includes bibliographical references (leaves 95-99). / List of figures --- p.iv / List of tables --- p.v / Abbreviations --- p.vi / Abstract --- p.vii / Chapter I. --- INTRODUCTION --- p.1 / Chapter 1. --- Inhibin a brief review --- p.1 / Chapter 1.1 --- "Definition and nomenclature, including related substances" --- p.1 / Chapter 1.2 --- Structure --- p.2 / Chapter 1.3 --- Historical background of the inhibin concept --- p.4 / Chapter 1.4 --- Actions of inhibin --- p.9 / Chapter 1.5 --- Control of inhibin production --- p.10 / Chapter 1.6 --- Measurement --- p.11 / Chapter 1.6.1 --- Immunoassay --- p.11 / Chapter 1.6.2 --- Bioassay --- p.13 / Chapter 1.6.2.1 --- In vivo methods --- p.13 / Chapter 1.6.2.2 --- In vitro methods --- p.14 / Chapter 1.7 --- Inhibin in clinical studies --- p.15 / Chapter 2. --- Project design --- p.17 / Chapter 2.1 --- Background --- p.17 / Chapter 2.2 --- Objectives --- p.20 / Chapter II. --- MATERIALS AND METHODS --- p.21 / Chapter 1. --- Materials --- p.21 / Chapter 1.1 --- Tracer preparation and purification --- p.21 / Chapter 1.2 --- Inhibin RIA --- p.21 / Chapter 1.3 --- Other immunoassays --- p.22 / Chapter 2. --- In-house inhibin RIA development --- p.22 / Chapter 2.1 --- The tracer preparation --- p.22 / Chapter 2.2 --- The radioimmunoassay --- p.25 / Chapter 2.3 --- Optimization of assay parameters --- p.26 / Chapter 2.3.1 --- Optimization of serum content and second antibody titre --- p.26 / Chapter 2.3.2 --- Verification of second antibodies' precipitating activity --- p.27 / Chapter 2. --- INHIBIN-EASIA --- p.27 / Chapter 3. --- Progesterone --- p.28 / Chapter 4. --- "Oestradiol, LH, and FSH" --- p.28 / Chapter III. --- RESULTS --- p.30 / Chapter Part I. --- In-house inhibin assay development --- p.30 / Chapter 1. --- Iodination and purification --- p.30 / Chapter 1.1 --- Step 1:Iodination followed by Sephadex column purification --- p.30 / Chapter 1.2 --- Step 2: Red A gel column purification --- p.33 / Chapter 1.3 --- Step 3: Sephadex column purification --- p.33 / Chapter 2. --- Inhibin RIA: Binding and antibody dilution curve experiment --- p.36 / Chapter 3.1 --- Verification of binding activity of the second antibody --- p.38 / Chapter 3.2 --- Optimization of serum content/ second antibody titre --- p.39 / Chapter 4. --- Discussion and conclusion --- p.41 / Chapter Part II. --- Hormone results of women undergoing in vitro fertilization --- p.42 / Chapter 1. --- Presentation of analytical results --- p.42 / Chapter 2. --- Comparison of hormone profiles of patients in the two GnRH agonist regimes --- p.43 / Chapter 2.1 --- Gonadotropins --- p.43 / Chapter 2.2 --- E2 --- p.56 / Chapter 2.3 --- Progesterone --- p.62 / Chapter 2.4 --- Inhibin --- p.68 / Chapter 3. --- Relationship between hormone --- p.74 / Chapter 3.1 --- Relationship between hormone changes --- p.74 / Chapter 3.2 --- Regression analysis --- p.87 / Chapter IV. --- DISCUSSSION --- p.89 / Chapter 1. --- Hormone profiles --- p.89 / Chapter 2. --- Hormone correlation --- p.91 / Chapter V. --- CONCLUSION --- p.94 / Chapter VI. --- REFERENCES --- p.95 / Chapter VIII. --- Appendix 1 Protocol for study on IVF and inhibin --- p.100 / Chapter IX. --- Appendix 2 Protocol for the management of IVF cycles --- p.101 / Chapter X. --- Appendix 3 Patients' results (table) --- p.103 / Chapter XI. --- Appendix 4 Patients ' results (graph) --- p.110

Introdução: O hermafroditismo verdadeiro, caracterizado pela demonstração histológica de tecido ovariano e testicular no mesmo indivíduo, responde por cerca de 5% dos casos de anomalia da diferenciação sexual. Como a variabilidade fenotípica é muito grande, desde mulheres com genitália externa normal até homens com genitália externa normal, passando por toda uma gama de apresentações intermediárias, torna-se impossível o diagnóstico baseado apenas em dados clínicos. A avaliação da presença de tecido testicular é bem estabelecida, mas não há teste para a demontração de tecido ovariano. A inibina A é produzida exclusivamente no ovário e é estimulada pelas gonadotrofinas. Objetivos: 1. Avaliar a efetividade do método de estimulação gonadal com a associação LH/FSH na demonstração de tecido ovariano; 2. Avaliar a eventual presença de tecido ovariano em pacientes com anomalias da diferenciação sexual através da dosagem sérica de Inibina A e de estradiol após estímulo gonadotrófico e; 3. Facilitar o diagnóstico de hermafroditismo verdadeiro antes da fase de exploração cirúrgica das gônadas. Métodos: Foram incluídos no estudo, dez pacientes com hiperplasia congênita de supra-renal, dez pacientes com criptorquidia unilateral isolada, treze pacientes com anomalia da diferenciação sexual sem etiologia definida e sete pacientes com hermafroditismo verdadeiro com diagnóstico histológico. Todos os pacientes foram submetidos a um teste de estímulo gonadotrófico, representado pela administração de gonadotrofina humana da menopausa (menotropina), que tem em sua composição LH e FSH, na dose de 150 UI de cada gonadotrofina, por via intramuscular, durante três dias subseqüentes. Dosagens de LH, FSH, estradiol, testosterora e inibina A foram realizadas antes (B), 24h após a primeira dose (A1) e 24 horas após a terceira dose (A2). Resultados: O LH não apresentou elevação significativa nos quatro grupos. O FSH elevou-se nos quatro grupos de forma progressiva e semelhante. O estradiol elevou-se significativamente nos grupos de pacientes com hiperplasia congênita das supra-renais (p=0,005) e de pacientes com hermafroditismo verdadeiro (p=0,031), enquanto a testosterona elevou-se nos grupos com criptorquidia isolada (p=0,027) e de pacientes com ambigüidade genital sem etiologia definida (p=0,028). A inibina A elevou-se significativamente nos grupos de pacientes com hiperplasia congênita das supra-renais (p=0,005) e com hermafroditismo verdadeiro (p=0,043). Conclusão: O teste de estímulo com LH e FSH mostrou-se útil para o diagnóstico da presença de tecido ovariano tanto em pacientes com hiperplasia congênita das supra-renais, como naqueles com hermafroditismo verdadeiro. / Introduction: True hermaphrodism (TH) is characterized by the presence of ovarian and testicular tissue in the same patient comprises 5% of the intersex cases. A large spectrum of phenotypical variation is observed, ranging from normal female genitalia to normal male genitalia, covering a wide range of intermediary presentations, it becomes very difficult to make the diagnosis of TH on clinical basis. The detection of testicular tissue is well stablished but there is no available test to demonstrate the presence of ovarian tissue. Objectives: 1. To evaluate the effectiveness of the LH/FSH gonadal stimulation in demonstrating ovarian functiom 2. To evaluate the presence of ovarian tissue in intersex patients under gonadotropic stimulation and 3. To make the TH diagnosis before the surgical procedure. Patients and Methods: Ten patients with congenital adrenal hyperplasia (CAH), 10 with unilateral cryptorchidism, 13 intersex patients with no defined etiology, and seven TH patients have been included in the study. All the patients had a gonadotropic stimulation test with human menopausal gonadotropin (menotropin-hMG),150 IU, intramuscular, for three consecutive days. LH, FSH, estradiol, testosterone, and Inhibin A were measured before (0 time), 24h after the first gonadotropin dose, and 24h after the third gonadotropin dose. Results: LH did not show any significant increase in the four groups studied. FSH increased in the four groups in a similar way. Estradiol increased in CAH pacients (p=0.005) and in TH patients (p=0,031), while testosterone increased in patients whit unilateral cryptorchidism (p=0.027) as well as in the intersex patients without defined etiology. Inhibin A levels increased in CAH patients (p=0.005) and in the TH patients (p=0.043). Conclusion: The LH/FSH stimulation test demonstrated to be a useful method to diagnose the presence of ovarian tissue in CAH patients as well as in TH patients, becoming an important tool to diagnose TH even before the surgical procedure and histologic studies of the gonads.

Objetivo: Avaliar a função das células testiculares de Sertoli em homens com lúpus eritematoso sistêmico (LES). Métodos: Trinta e quatro pacientes consecutivos foram prospectivamente selecionados para avaliar a função das células testiculares de Sertoli pelos níveis séricos da inibina B. Características clínicas, tratamento, análises dos espermatozóides, avaliação urológica, ultrasonografia testicular, hormônios e anticorpos anti-espermatozóides foram avaliados. Resultados: Os pacientes foram subdivididos em dois grupos: níveis séricos reduzidos (Grupo 1, n=8) e níveis séricos normais (Grupo 2, n=26) de inibina B. As medianas da concentração média de espermatozóides (p=0,024), da contagem total de espermatozóides (p=0,023) e da contagem total de espermatozóides móveis (p=0,025) foram menores no Grupo 1. Os níveis de inibina B foram positivamente correlacionados com a concentração de espermatozóides (r=0,343) e contagem total de espermatozóides móveis (r=0,357), e negativamente correlacionados com FSH (r=0,699) e LH (r=0,397). A mediana de inibina B sérica foi menor nos pacientes com LES tratados com pulsoterapia com ciclofosfamida endovenosa (CICIV) comparada aos não tratados com este medicamento (p=0,031). Avaliação dos 26 pacientes com LES e níveis normais de inibina B e FSH revelou que a mediana da relação inibina B/FSH foi menor nos pacientes lúpicos com oligozoospermia comparada aos pacientes com normozoospermia (p=0,004). Esta relação também foi menor nos pacientes com LES tratados com CICIV do que naqueles sem esta terapia (p=0,04). Conclusão: Este é o primeiro estudo que identificou uma alta freqüência de disfunção das células testiculares de Sertoli em homens com LES associada a anormalidades dos espermatozóides. Outros estudos prospectivos são necessários para determinar se os níveis séricos de inibina e a relação inibina B/FSH serão um marcador útil e precoce de toxicidade pela CICIV neste pacientes / Objective: To assess the testicular Sertoli cell function in male SLE patients. Methods: 34 consecutive patients were prospectively selected to evaluate the testicular Sertoli cell function by serum inhibin B levels. Clinical features, treatment, semen analysis, urologic evaluation, testicular ultrasound, hormones, and antisperm antibodies were determined. Results: Patients were subdivided into two Groups: low serum inhibin B (Group 1, n=8) and normal levels (Group 2, n=26). The medians of sperm concentration (p=0.024), total sperm count (p=0.023) and total motile sperm count (p=0.025) were lower in Group 1. Inhibin B levels were positively correlated with sperm concentration (r=0.343), total motile sperm count (r=0.357), and negatively correlated with FSH (r=0.699) and LH (r=0.397). The median serum inhibin B was lower in SLE patients treated with intravenous cyclophosphamide pulsetherapy (IVCYC) compared to those without this therapy (p=0.031). Further evaluation of the 26 SLE patients with normal inhibin B and FSH levels revealed that medians of inhibin B/FSH ratio were lower in SLE patients with oligozoospermia compared to normozoospermia (p=0.004). This ratio was also lower in SLE patients treated with IVCYC than those without this therapy (p=0.04). Conclusions: This is the first study to identify a high frequency of testicular Sertoli cell dysfunction in male SLE associated with sperm abnormalities. Further prospective studies are necessary to determine if inhibin levels and inhibin B/FSH ratio will be an earlier and useful marker of IVCYC toxicity in these patients

Objetivo: Avaliar a função das células testiculares de Sertoli em homens com lúpus eritematoso sistêmico (LES). Métodos: Trinta e quatro pacientes consecutivos foram prospectivamente selecionados para avaliar a função das células testiculares de Sertoli pelos níveis séricos da inibina B. Características clínicas, tratamento, análises dos espermatozóides, avaliação urológica, ultrasonografia testicular, hormônios e anticorpos anti-espermatozóides foram avaliados. Resultados: Os pacientes foram subdivididos em dois grupos: níveis séricos reduzidos (Grupo 1, n=8) e níveis séricos normais (Grupo 2, n=26) de inibina B. As medianas da concentração média de espermatozóides (p=0,024), da contagem total de espermatozóides (p=0,023) e da contagem total de espermatozóides móveis (p=0,025) foram menores no Grupo 1. Os níveis de inibina B foram positivamente correlacionados com a concentração de espermatozóides (r=0,343) e contagem total de espermatozóides móveis (r=0,357), e negativamente correlacionados com FSH (r=0,699) e LH (r=0,397). A mediana de inibina B sérica foi menor nos pacientes com LES tratados com pulsoterapia com ciclofosfamida endovenosa (CICIV) comparada aos não tratados com este medicamento (p=0,031). Avaliação dos 26 pacientes com LES e níveis normais de inibina B e FSH revelou que a mediana da relação inibina B/FSH foi menor nos pacientes lúpicos com oligozoospermia comparada aos pacientes com normozoospermia (p=0,004). Esta relação também foi menor nos pacientes com LES tratados com CICIV do que naqueles sem esta terapia (p=0,04). Conclusão: Este é o primeiro estudo que identificou uma alta freqüência de disfunção das células testiculares de Sertoli em homens com LES associada a anormalidades dos espermatozóides. Outros estudos prospectivos são necessários para determinar se os níveis séricos de inibina e a relação inibina B/FSH serão um marcador útil e precoce de toxicidade pela CICIV neste pacientes / Objective: To assess the testicular Sertoli cell function in male SLE patients. Methods: 34 consecutive patients were prospectively selected to evaluate the testicular Sertoli cell function by serum inhibin B levels. Clinical features, treatment, semen analysis, urologic evaluation, testicular ultrasound, hormones, and antisperm antibodies were determined. Results: Patients were subdivided into two Groups: low serum inhibin B (Group 1, n=8) and normal levels (Group 2, n=26). The medians of sperm concentration (p=0.024), total sperm count (p=0.023) and total motile sperm count (p=0.025) were lower in Group 1. Inhibin B levels were positively correlated with sperm concentration (r=0.343), total motile sperm count (r=0.357), and negatively correlated with FSH (r=0.699) and LH (r=0.397). The median serum inhibin B was lower in SLE patients treated with intravenous cyclophosphamide pulsetherapy (IVCYC) compared to those without this therapy (p=0.031). Further evaluation of the 26 SLE patients with normal inhibin B and FSH levels revealed that medians of inhibin B/FSH ratio were lower in SLE patients with oligozoospermia compared to normozoospermia (p=0.004). This ratio was also lower in SLE patients treated with IVCYC than those without this therapy (p=0.04). Conclusions: This is the first study to identify a high frequency of testicular Sertoli cell dysfunction in male SLE associated with sperm abnormalities. Further prospective studies are necessary to determine if inhibin levels and inhibin B/FSH ratio will be an earlier and useful marker of IVCYC toxicity in these patients

Introdução: O hermafroditismo verdadeiro, caracterizado pela demonstração histológica de tecido ovariano e testicular no mesmo indivíduo, responde por cerca de 5% dos casos de anomalia da diferenciação sexual. Como a variabilidade fenotípica é muito grande, desde mulheres com genitália externa normal até homens com genitália externa normal, passando por toda uma gama de apresentações intermediárias, torna-se impossível o diagnóstico baseado apenas em dados clínicos. A avaliação da presença de tecido testicular é bem estabelecida, mas não há teste para a demontração de tecido ovariano. A inibina A é produzida exclusivamente no ovário e é estimulada pelas gonadotrofinas. Objetivos: 1. Avaliar a efetividade do método de estimulação gonadal com a associação LH/FSH na demonstração de tecido ovariano; 2. Avaliar a eventual presença de tecido ovariano em pacientes com anomalias da diferenciação sexual através da dosagem sérica de Inibina A e de estradiol após estímulo gonadotrófico e; 3. Facilitar o diagnóstico de hermafroditismo verdadeiro antes da fase de exploração cirúrgica das gônadas. Métodos: Foram incluídos no estudo, dez pacientes com hiperplasia congênita de supra-renal, dez pacientes com criptorquidia unilateral isolada, treze pacientes com anomalia da diferenciação sexual sem etiologia definida e sete pacientes com hermafroditismo verdadeiro com diagnóstico histológico. Todos os pacientes foram submetidos a um teste de estímulo gonadotrófico, representado pela administração de gonadotrofina humana da menopausa (menotropina), que tem em sua composição LH e FSH, na dose de 150 UI de cada gonadotrofina, por via intramuscular, durante três dias subseqüentes. Dosagens de LH, FSH, estradiol, testosterora e inibina A foram realizadas antes (B), 24h após a primeira dose (A1) e 24 horas após a terceira dose (A2). Resultados: O LH não apresentou elevação significativa nos quatro grupos. O FSH elevou-se nos quatro grupos de forma progressiva e semelhante. O estradiol elevou-se significativamente nos grupos de pacientes com hiperplasia congênita das supra-renais (p=0,005) e de pacientes com hermafroditismo verdadeiro (p=0,031), enquanto a testosterona elevou-se nos grupos com criptorquidia isolada (p=0,027) e de pacientes com ambigüidade genital sem etiologia definida (p=0,028). A inibina A elevou-se significativamente nos grupos de pacientes com hiperplasia congênita das supra-renais (p=0,005) e com hermafroditismo verdadeiro (p=0,043). Conclusão: O teste de estímulo com LH e FSH mostrou-se útil para o diagnóstico da presença de tecido ovariano tanto em pacientes com hiperplasia congênita das supra-renais, como naqueles com hermafroditismo verdadeiro. / Introduction: True hermaphrodism (TH) is characterized by the presence of ovarian and testicular tissue in the same patient comprises 5% of the intersex cases. A large spectrum of phenotypical variation is observed, ranging from normal female genitalia to normal male genitalia, covering a wide range of intermediary presentations, it becomes very difficult to make the diagnosis of TH on clinical basis. The detection of testicular tissue is well stablished but there is no available test to demonstrate the presence of ovarian tissue. Objectives: 1. To evaluate the effectiveness of the LH/FSH gonadal stimulation in demonstrating ovarian functiom 2. To evaluate the presence of ovarian tissue in intersex patients under gonadotropic stimulation and 3. To make the TH diagnosis before the surgical procedure. Patients and Methods: Ten patients with congenital adrenal hyperplasia (CAH), 10 with unilateral cryptorchidism, 13 intersex patients with no defined etiology, and seven TH patients have been included in the study. All the patients had a gonadotropic stimulation test with human menopausal gonadotropin (menotropin-hMG),150 IU, intramuscular, for three consecutive days. LH, FSH, estradiol, testosterone, and Inhibin A were measured before (0 time), 24h after the first gonadotropin dose, and 24h after the third gonadotropin dose. Results: LH did not show any significant increase in the four groups studied. FSH increased in the four groups in a similar way. Estradiol increased in CAH pacients (p=0.005) and in TH patients (p=0,031), while testosterone increased in patients whit unilateral cryptorchidism (p=0.027) as well as in the intersex patients without defined etiology. Inhibin A levels increased in CAH patients (p=0.005) and in the TH patients (p=0.043). Conclusion: The LH/FSH stimulation test demonstrated to be a useful method to diagnose the presence of ovarian tissue in CAH patients as well as in TH patients, becoming an important tool to diagnose TH even before the surgical procedure and histologic studies of the gonads.