Analysis of anti-Mullerian hormone (AMH) and its receptor (AMHR2) genes in patients with persistent Mullerian duct syndrome

Objective: To screen for mutations in AMH and AMHR2 genes in patients with persistent Mullerian duct syndrome (PMDS). Patients and method: Genomic DNA of eight patients with PMDS was obtained from peripheral blood leukocytes. Directed sequencing of the coding regions and the exon-intron boundaries of AMH and AMHR2 were performed. Results: The AMH mutations p.Arg95*, p.Arg123Trp, c.556-2A>G, and p. Arg502Leu were identified in five patients; and p.Gly323Ser and p.Arg407* in AMHR2 of two individuals. In silico analyses of the novel c.556-2A>G, p.Arg502Leu and p.Arg407* mutations predicted that they were harmful and were possible causes of the disease. Conclusion: A likely molecular etiology was found in the eight evaluated patients with PMDS. Four mutations in AMH and two in AMHR2 were identified. Three of them are novel mutations, c.556-2A>G, and p. Arg502Leu in AMH; and p.Gly323Ser in AMHR2. Arq Bras Endocrinol Metab. 2012;56(8):473-8Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq) [305743/2011-2, 314392/2009-2, 302084/2009-6]Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (Fapesp) [06/50999-0]Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP

Pesquisa de mutações nos genes do hormônio antiMülleriano (AMH) e do seu receptor (AMHR2) em pacientes com síndrome de persistência dos ductos Müllerianos

OBJECTIVE: To screen for mutations in AMH and AMHR2 genes in patients with persistent Müllerian duct syndrome (PMDS). PATIENTS AND METHOD: Genomic DNA of eight patients with PMDS was obtained from peripheral blood leukocytes. Directed sequencing of the coding regions and the exon-intron boundaries of AMH and AMHR2 were performed. RESULTS: The AMH mutations p.Arg95*, p.Arg123Trp, c.556-2A>G, and p.Arg502Leu were identified in five patients; and p.Gly323Ser and p.Arg407* in AMHR2 of two individuals. In silico analyses of the novel c.556-2A>G, p.Arg502Leu and p.Arg407* mutations predicted that they were harmful and were possible causes of the disease. CONCLUSION: A likely molecular etiology was found in the eight evaluated patients with PMDS. Four mutations in AMH and two in AMHR2 were identified. Three of them are novel mutations, c.556-2A>G, and p.Arg502Leu in AMH; and p.Gly323Ser in AMHR2. Arq Bras Endocrinol Metab. 2012;56(8):473-8OBJETIVO: Analisar os genes AMH e AMHR2 em indivíduos com síndrome de persistência dos ductos de Müller (SPDM). PACIENTES E MÉTODO: Amostras de DNA genômico de oito pacientes com SPDM foram obtidas de leucócitos de sangue periférico. Sequenciamento direto da região codificadora e das áreas intrônicas próximas aos éxons dos genes AMH e AMHR foi realizado. RESULTADOS: As mutações p.Arg95*, p.Arg123Trp, c.556-2A>G e p.Arg502Leu no gene AMH foram identificadas em cinco pacientes e as mutações p.Gly323Ser e p.Arg407* no gene AMHR2, em dois indivíduos. As análises in silico das mutações c.556-2A>G, p.Arg502Leu e p.Arg407*, não descritas anteriormente na literatura, previram que elas são deletérias e possivelmente a causa da doença. CONCLUSÃO: Uma provável etiologia molecular foi encontrada nos oito pacientes portadores de SPDM avaliados. No gene do AMH foram identificadas quatro mutações e no AMHR2, duas mutações. Três das seis mutações encontradas são mutações novas, c.556-2A>G e p.Arg502Leu no gene AMH; e p.Gly323Ser no AMHR2. Arq Bras Endocrinol Metab. 2012;56(8):473-8473478Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Analysis of anti-Mullerian hormone (AMH) and its receptor (AMHR2) genes in patients with persistent Mullerian duct syndrome

Objective: To screen for mutations in AMH and AMHR2 genes in patients with persistent Mullerian duct syndrome (PMDS). Patients and method: Genomic DNA of eight patients with PMDS was obtained from peripheral blood leukocytes. Directed sequencing of the coding regions and the exon-intron boundaries of AMH and AMHR2 were performed. Results: The AMH mutations p.Arg95*, p.Arg123Trp, c.556-2A>G, and p. Arg502Leu were identified in five patients; and p.Gly323Ser and p.Arg407* in AMHR2 of two individuals. In silico analyses of the novel c.556-2A>G, p.Arg502Leu and p.Arg407* mutations predicted that they were harmful and were possible causes of the disease. Conclusion: A likely molecular etiology was found in the eight evaluated patients with PMDS. Four mutations in AMH and two in AMHR2 were identified. Three of them are novel mutations, c.556-2A>G, and p. Arg502Leu in AMH; and p.Gly323Ser in AMHR2. Arq Bras Endocrinol Metab. 2012;56(8):473-8Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq) [305743/2011-2, 314392/2009-2, 302084/2009-6]Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (Fapesp) [06/50999-0]Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP

Avaliação dos plexos mioentericos e do transito intestinal em ratos submetidos a lesão por isquemia e reperfusão do territorio da arteria mesenterica superior

Orientador: Joaquim Murray Bustorff-SilvaDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias MedicasResumo: Alterações histológicas precoces causadas por enterocolite necrosante já foram exaustivamente estudadas em modelos animais. No entanto, poucas informações sobre os efeitos tardios da doença sobre a motilidade intestinal podem ser encontradas na literatura atual. No presente estudo, trinta e dois ratos Wistar machos recém-desmamados pesando entre 58 e 103 gramas foram distribuídos ao acaso em quatro grupos: Controle (não operados), Sham (laparotomia), Isquemia por 30 minutos (laparotomia e isquemia da artéria mesentérica superior por 30 minutos) e Isquemia por 45 minutos (laparotomia e isquemia da artéria mesentérica superior por 45 minutos). A motilidade intestinal no período pós-operatório foi medida indiretamente através da medida do peso seco do total de fezes obtidas em 24 horas nos 3°, 7°, 14° e 210 dias após a cirurgia. Segmentos de duodeno, jejuno e íleo foram examinados à microscopia de luz para observação de alterações ocorridas nos plexos mioentéricos. Três semanas após a isquemia, os neurômos dos plexos mioentéricos apresentaram vacuolização citoplasmática, e seus núcleos apresentaram irregularidade no f:Ontomo e sinais degenerativos. Houve mudanças significativas na motilidade intestinal nos animais submetidos a 45 minutos de isquemia mesentérica, mas essas mudanças não provocaram alterações significativas no crescimento final dos animais. Os resultados sugerem que o processo de isquemia-reperfusão intestinal causa danos aos neurômos dos plexos mioentéricos. Nas condições referidas no presente estudo, essas mudanças foram suficientes para induzir alterações na motilidade, mas isso não afetou a função intestinalAbstract: Background/Purpose: Early histological changes induced by necrotizing enterocolitis have been extensively studied using animal models. However, infonnation regarding late effects on intestinal motility are lacking. The aim of this study was to investigate the late effects of ischemia-reperfusion on myenteric pIexus histology and intestinal motility.Materiais and Methods: Thirty-two post-weaning male mice weighing between 58 and l03g were divided randomly into 4 groups: Control (unoperated), Sbam(celiotomy), 30 min Ischemia(celiotomy and superior mesenteric artery ischemia for 30 minutes) and 45 min Ischemia(celiotomy and superior mesenteric artery ischemia for 45 minutes). Postoperative intestinal motility was assessed by weighing total fecal output over 24 hours on the 3rd, 7th, 14th and 21 st day afier surgery. Segments of duodenum, jejunum and ileum were examined at light microscopy for changes in the myenteric plexus.Results: At three weeks afier ischemia, the myenteric neurons appeared, in light microscopy, spongy or foamy, containing many vacuoIes in their cytoplasm. The neuronal nucIeus became irregular, with degenerative signs. There were significantly changes in intestinal motility in the animals groups submitted to intestinal ischemia, but thest: changes did not affect the animal overall growth.Conclusions: The results suggest that intestinal ischemia-reperfusion causes late neuronal damage. Within that conditions of the present study, these changes were sufficient to induce alterations in intestinal motility, but this did not affect intestinal functionMestradoCirurgiaMestre em Cirurgi