Extração de DNA de materiais de arquivo e fontes escassas para utilização em reação de polimerização em cadeia (PCR)

Este trabalho visou a comparação de cinco métodos diferentes de extração de DNA de materiais de arquivo (tecidos incluídos em parafina, esfregaços de sangue periférico - corados e não corados com Leishman, lâminas com mielogramas, gotas de sangue em Guthrie Card) e de fontes escassas (células bucais, um e três bulbos capilares e 2 mL de urina), para que fossem avaliadas a facilidade de aplicação e a facilidade de amplificação deste DNA pela técnica da reação de polimerização em cadeia (PCR). Os métodos incluíram digestão por proteinase K, seguida ou não por purificação com fenol/clorofórmio; Chelex 100® (BioRad); Insta Gene® (BioRad) e fervura em água estéril. O DNA obtido foi testado para amplificação de três fragmentos gênicos: Brain-derived neutrophic factor (764 pb), Factor V Leiden (220 pb) e Abelson (106 pb). de acordo com o comprimento do fragmento gênico estudado, da fonte potencial de DNA e do método de extração utilizado, os resultados caracterizaram o melhor caminho para padronização de procedimentos técnicos a serem incluídos no manual de Procedimentos Operacionais Padrão do Laboratório de Biologia Molecular do Hemocentro - HC - Unesp - Botucatu.

Bone marrow necrosis related to paracoccidioidomycosis: the first eight cases identified at autopsy

To report the first eight bone marrow necrosis (BMN) cases related to paracoccidioidomycosis (PCM) from patient autopsies with well-documented bone marrow (BM) histology and cytology.A retrospective evaluation was performed on BM specimens from eight autopsied patients from Botucatu University Hospital with PCM-related BMN. Relevant BMN literature was searched and analysed.All eight patients had acute PCM. Six had histological only (biopsies) and two cytological only (smears) specimens. Five biopsy specimens revealed severe and one mild coagulation patterned necrotic areas. Five had osteonecrosis. The cytological specimens also showed typical BMN patterns. Paracoccidioides brasiliensis yeast forms were visible within necrotic areas in all cases.

Proteção, promoção e apoio ao aleitamento materno em um hospital universitário

Estudo de natureza quantitativa descritiva em um Hospital Universitário do Estado de São Paulo, teve como objetivo avaliar a promoção, proteção e apoio ao aleitamento materno através da verificação do cumprimento dos dez passos para o sucesso do aleitamento de acordo com a iniciativa Hospital Amigo da Criança, proposta pelo Ministério da Saúde. A avaliação revela práticas facilitadoras da amamentação, no entanto, faz-se necessário algumas mudanças nas rotinas hospitalares para a efetivação dos dez passos. em média, 79% das respostas foram afirmativas para o cumprimento dos passos, o que está próximo aos 80% preconizado. No entanto, se analisarmos as respostas para cada passo, identificamos pontos para serem melhorados. Chamou-nos a atenção o fato de que a prática de colocar o recém-nascido junto à mãe logo após o nascimento, permitindo a amamentação na primeira meia hora, foi relatada por 100% das mães entrevistadas que tiveram parto normal e por 80% daquelas que tiveram parto cesárea. Investimentos e mudanças em direção ao cumprimento dos dez passos contribuirão para um melhor atendimento à população e para a formação de profissionais amigos da criança.

Association between CD31 expression and histopathologic features in stage IB squamous cell carcinoma of the cervix

This study was undertaken to evaluate the association between the expression of CD31 in the tumor and the histopathologic findings in patients with carcinoma of the cervix. This study included prospectively 30 women, aged 46.6 +/- 10.7 years, with stage IB squamous cell carcinoma of the cervix submitted to radical hysterectomy from November 2001 to September 2002. Samples from the tumor were taken and immunohistochemically evaluated by a monoclonal antibody for CD31. Clinicopathologic characteristics such as stage, tumor size, grade of differentiation, lymphatic vascular space invasion (LVSI), parametrial involvement, and status of pelvic lymph nodes were also recorded. The clinical stage (FIGO) was IB1 in 22 patients (73.3%) and IB2 in 8 patients (26.7%). The expression of CD31 was significantly associated with tumor size and the presence of LVSI, but not with grade of differentiation and vaginal or parametrial involvement (P= 0.03, P= 0.032, P= 0.352, P= 0.208, and P= 0.242, respectively). on univariate analysis, the presence of pelvic lymph node metastasis was influenced by LVSI (P= 0.003) and CD31 expression (P= 0.032). However, on multivariate analysis, the presence of LVSI (P= 0.007) was the only independent predictor of pelvic lymph node metastasis. The CD31 expression in tumor is significantly associated with LVSI and tumor size in patients with early-stage squamous cell carcinoma of the cervix.

Cervical Lactobacillus and leukocyte infiltration in preterm premature rupture of membranes

Objectives: To evaluate the possible association between microorganisms present in the cervical secretions and amniotic fluid of pregnant women with preterm premature rupture of membranes (PPROM), and histologic chorioamnionitis. Methods: Thirty-seven pregnant women with PPROM and 21 healthy pregnant women were studied. Secretions from the cervical canal and amniotic fluid were collected to isolate microorganisms present in the genital tract. Cervical smears were Gram stained and evaluated microscopically. At delivery, chorioamniotic membranes were collected for histopathologic analysis. Results: Microscopic examination of the cervical secretion smears obtained from the PPROM group showed a low rate of Lactobacillus species, large numbers of leukocytes, and a wide diversity of microorganisms compared with the control group. The PPROM group presented an 80% rate of chorioamnionitis. Staphylococcus aureus isolation in cervical secretion was associated with intense inflammatory infiltrate in the membranes and might play a role in the pathogenesis of PPROM. Conclusions: the low colonization of cervical flora by Lactobacillus species associated with an intense leukocyte infiltrate detected in Gram-stained cervical smears can be considered a rapid method of detecting chorioamnionitis in pregnant women with PPROM. (C) 2003 International Federation of Gynecology and Obstetrics. Published by Elsevier B.V. Ireland Ltd. All rights reserved.

Prova de Trabalho de Parto Após uma Cesárea Anterior

Objetivo: estudar a validade da prova de trabalho de parto (PTP) em gestantes com uma cesárea anterior. Métodos: estudo retrospectivo, tipo coorte, incluindo 438 gestantes com uma cesárea anterior ao parto em estudo e seus 450 recém-nascidos (RN), divididas em dois grupos - com e sem PTP. O tamanho amostral mínimo foi de 121 gestantes/grupo. Considerou-se variável independente a PTP e as dependentes relacionaram-se à ocorrência de parto vaginal e à freqüência de complicações maternas e perinatais. Foram efetuadas análises uni e multivariada, respectivamente. A comparação entre as freqüências (%) foi analisada pelo teste do qui-quadrado (chi²) com significância de 5% e regressão logística com cálculo do odds ratio (OR) e do intervalo de confiança a 95% (IC95%). Resultados: a PTP associou-se a 59,2% de partos vaginais. Foi menos indicada nas gestantes com mais de 40 anos (2,7% vs 6,5%) e nas portadoras de doenças associadas e complicações da gravidez: síndromes hipertensivas (7,0%) e hemorragias de 3º trimestre (0,3%). A PTP não se relacionou às complicações maternas e perinatais. As gestantes que tiveram o parto por cesárea, independente da PTP, apresentaram maior risco de complicações puerperais (OR = 3,53; IC95% = 1,57-7,93). A taxa de mortalidade perinatal foi dependente do peso do RN e das malformações fetais e não se relacionou à PTP. Ao contrário, as complicações respiratórias foram mais freqüentes nos RN de mães não testadas quanto à PTP (OR = 1,92; IC95% = 1,20-3,07). Conclusões: os resultados comprovaram que a PTP em gestantes com uma cesárea anterior é estratégia segura - favoreceu o parto vaginal em 59,2% dos casos e não interferiu com a morbimortalidade materna e perinatal. Portanto, é recurso que deve ser estimulado.

Efeito da via de parto sobre a força muscular do assoalho pélvico

OBJETIVO: analisar a influência da via de parto sobre a força muscular do assoalho pélvico (FM-AP). MÉTODOS: estudo clínico de corte transversal, para avaliar a FM-AP pelo teste da avaliação da força do assoalho pélvico (AFA) e uso do perineômetro em primíparas, entre 20-30 anos de idade, 4-6 meses pós-parto. A contração, medida pelos dois testes, foi classificada em: zero - ausência, um - leve, dois - moderada e três - normal, sustentada por 6 segundos. Avaliaram-se 94 mulheres, entre 20 e 30 anos, divididas em três grupos: pós-parto vaginal (n=32); pós-cesárea (n=32) e nulíparas (n=30). A variável independente foi a via de parto e a dependente, a FM-AP. A comparação entre os graus de contração foi realizada pelo teste de Kruskal-Wallis e o teste de Dunn para comparações múltiplas; a influência da via de parto pelo teste chi2, o risco relativo (RR) para alteração da FM-AP e o coeficiente kappa para avaliar equivalência entre os testes. RESULTADOS: a mediana e 1º e 3º quartil da FM-AP foram menores (p=0,01) pós-parto vaginal (2,0;1-2) e intermediários pós-cesárea (2,0; 2-3) em relação às nulíparas (3,0;2-3), tanto analisadas pelo AFA como pelo perineômetro. Aumentou o RR de exame alterado pós-parto vaginal (RR=2,5; IC 95%: 1,3-5,0; p=0,002); (RR=2,3; IC 95%: 1,2-4,3; p=0,005) e pós-cesárea (RR=1,5; IC 95%: 0,94-2,57; p=0,12); (RR=1,3; IC 95%: 0,85-2,23; p=0,29) pelo PFSE e perineômetro, respectivamente. CONCLUSÕES: o parto vaginal diminuiu a força muscular do AP de primíparas quando comparado com os casos submetidos à cesárea e com as nulíparas.

Prevalence and Risk Factors of Chlamydia trachomatis Cervicitis in Pregnant Women at the Genital Tract Infection in Obstetrics Unit Care at Botucatu Medical School, São Paulo State University - UNESP, Brazil

Objective. To evaluate the prevalence of and risk factors for Chlamydia trachomatis cervicitis in pregnant women seen at the Genital Tract Infection in Obstetrics Unit Care in Botucatu Medical School, São Paulo State University - UNESP.Materials and Methods. Between June 2006 and February 2008, 101 pregnant women were included in this study. During the gynecologic examination, cervical secretions were collected using cytobrush Plus GT (CooperSurgical Inc) to assess C. trachomatis using polymerase chain reaction. Vaginal flora were examined by Gram stain, and socio-demographic data were extracted from medical records.Results. of the 101 patients, 26 (25.7%) were positive for C. trachomatis. The median age of the infected group was 24 years (range = 13-40 y), and 48.5% of them had abnormal vaginal flora. The presence of chlamydial infection was associated with smoking (odds ratio [OR] = 2.67, 95% confidence interval [CI] = 1.01-7.19), residing in a city with fewer than 100,000 inhabitants (OR = 2.86, 95% CI = 1.03-7.94), presence of condyloma acuminatum (p = .03), and presence of discreet inflammation on Pap smear (p = .02).Conclusions. The prevalence of C. trachomatis is high in pregnant women seen at the Genital Infection Unit Care, UNESP, and is related to many risk factors. Therefore, its screening is extremely important in reducing obstetrical and neonatal complications.

Detection of Streptococcus agalactiae colonization in pregnant women by using combined swab cultures: cross-sectional prevalence study

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Mola hidatiforme completa e eclâmpsia: relato de caso

O desenvolvimento de pré-eclâmpsia ou eclâmpsia antes da 20ª semana deve levar à suspeita de mola hidatiforme. Descrevemos um caso de mola hidatiforme completa (MHC) e eclâmpsia concomitante em paciente com 20 anos que apresentava sangramento genital, anemia, tamanho uterino excessivo e cistos de ovário, associados a hipertensão arterial e proteinúria. Os níveis de b-hCG estavam elevados e a função tiroidiana, alterada. A ultra-sonografia mostrou-se compatível com MHC. Após o esvaziamento uterino apresentou cefaléia e alterações visuais, seguidas por convulsões tônico-clônicas que cessaram com sulfato de magnésio hepta-hidratado a 50%. No seguimento pós-molar foi diagnosticado tumor trofoblástico gestacional (TTG) prontamente tratado com quimioterapia. A associação de MHC e eclâmpsia determina esvaziamento uterino imediato e seguimento pós-molar rigoroso, pelo risco aumentado de desenvolvimento de TTG.

Medical students' personal choice for mode of delivery in Santa Catarina, Brazil: a cross-sectional, quantitative study

Background: The increase in overall rates of cesarean sections (CS) in Brazil causes concern and it appears that multiple factors are involved in this fact. In 2009, undergraduate students in the first and final years of medical school at the University of Santa Catarina answered questionnaires regarding their choice of mode of delivery. The aim of the study was to evaluate whether the education process affects decision-making regarding the waay of childbirth preferred by medical students.Methods: A cross-sectional, quantitative study was conducted based on data obtained from questionnaires applied to medical students. The questions addressed four different scenarios in childbirth, as follows: under an uneventful pregnancy; the mode of delivery for a pregnant woman under their care; the best choice as a healthcare manager and lastly, choosing the birth of their own child. For each circumstance, there was an open question to explain their choice.Results: A total of 189 students answered the questionnaires. For any uneventful pregnancy and for a pregnant woman under their care, 8.46% of the students would opt for CS. As a healthcare manager, only 2.64% of the students would recommend CS. For these three scenarios, the answers of the students in the first year did not differ from those given by students in the sixth year. In the case of the student's own or a partner's pregnancy, 41.4% of those in the sixth year and 16.8% of those in the first year would choose a CS. A positive association was found between being a sixth year student and a personal preference for CS according to logistic regression (OR = 2.91; 95%CI: 1.03-8.30). Pain associated with vaginal delivery was usually the reason for choosing a CS.Conclusions: A higher number of sixth year students preferred a CS for their own pregnancy (or their partner's) compared to first year students. Pain associated with vaginal delivery was the most common reason given for haven chosen a CS. The students' preference for childbirth changed over time during their graduation in favor of cesarean sections. This finding deserves considerable attention when structuring medical education in Obstetrics.

Molar Pregnancy in Adolescents

OBJECTIVE: To describe the clinical presentation of hydatidiform molar pregnancy in women under the age of 20 years. In addition, we sought to understand if this adolescent population manifests differences in clinical factors compared to an adult population that may affect outcome.STUDY DESIGN: We used a database from the New England Trophoblastic Disease Center to analyze clinical data from all women followed for molar pregnancy between 1970 and 2009 with complete follow-up information. This population was stratified by age and clinical parameters including presenting signs, molar histology and development of gestational trophoblastic neoplasia (GTN). Univariable and multivariable logistic regression was employed to discern clinical factors that associated with adolescent age. The Partners Human Research Committee approved this study.RESULTS: We identified 1,494 women diagnosed with hydatidiform mole (HM), of which 220 (14.7%) were adolescents defined as age <20 years. The most common presenting clinical signs were vaginal bleeding and an enlarged uterus compared to dates. Median gestational age at diagnosis was 13.4 weeks, not different from that in the adult population. Similarly, no difference in presenting human chorionic gonadotropin was observed between the adult and adolescent populations. Adolescents presented with a significant overrepresentation of complete mole (86% vs. 75%, p < 0.001) compared to adults. Complete mole was associated with a heightened risk of developing GTN (OR 2.6, 95% CI 1.9-3.5), and despite the association of complete mole with young maternal age, univariable analysis showed no difference in the rate of GTN observed between adolescents and adults (24% vs. 30%, p = 0.08). Multivariable analysis controlling for molar histology demonstrated that adolescent age was associated with a decreased risk of GTN (hazard ratio 0.67, 95% CI 0.48 0.93).CONCLUSION: Adolescents account for a substantial proportion of the population with HM. They commonly present with vaginal bleeding. Though this population develops a complete mole with a higher frequency than adults, adolescents appear to have a significantly decreased risk of developing GTN. (J Reprod Med 2012; 57:225-230)

Hydatidiform Mole in Ectopic Pregnancy Clinical, Imaging, Pathological and Immunohistochemical Characteristics

OBJECTIVE: To determine the clinical, pathological, immunohistochemical and imaging characteristics of hydatidiform mole in ectopic pregnancy (HMEP) in all the cases admitted to the Department of Obstetrics and Gynecology, University Hospital of Caracas (HUC), Central University of Venezuela.STUDY DESIGN: Retrospective and comparative study, based on clinical records review of 2 groups: 10 cases with a diagnosis of HMEP and 20 cases with intrauterine hydatidiform mole (IUHM) admitted to the Obstetrics and Gynecology Department of HUC from 1996 to 2010. Clinical, pathological, immunohistochemical and imaging features were analyzed.RESULTS: The prevalence of HMEP in this study was 0.14:1,000 pregnancies; in this group the mean age was 28.8 years, and the mean gestational age at admission was 8.6 weeks. Both groups (HMEP and IUHM) were comparable in these last variables. Abdominal pain and genital bleeding were the most common clinical symptoms in the HMEP group, while it was vaginal bleeding in the IUHM group. Ultrasound findings were similar to those traditionally described in nonmolar ectopic pregnancy. Histology and immunohistochemistry showed that all cases of HMEP were partial mole.CONCLUSION: Although in this study the prevalence of HMEP was high, the size of the sample limits definitive conclusions. This study concludes that all cases of HMEP are partial mole. (J Reprod Med 2012;57:329-332)

A multidisciplinary program of preparation for childbirth and motherhood: maternal anxiety and perinatal outcomes

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Efficacy of the motile sperm organelle morphology examination (MSOME) in predicting pregnancy after intrauterine insemination

Background: Although the motile sperm organelle morphology examination (MSOME) was developed merely as a selection criterion, its application as a method for classifying sperm morphology may represent an improvement in the evaluation of semen quality. The aim of this study was to determine the prognostic value of normal sperm morphology using MSOME with regard to clinical pregnancy (CP) after intrauterine insemination (IUI).Methods: A total of 156 IUI cycles that were performed in 111 couples were prospectively analysed. Each subject received 75 IU of recombinant FSH every second day from the third day of the cycle. Beginning on the 10th day of the cycle, follicular development was monitored by vaginal ultrasound. When one or two follicles measuring at least 17 mm were observed, recombinant hCG was administered, and IUI was performed 12-14 h and 36-40 h after hCG treatment. Prior to the IUI procedure, sperm samples were analysed by MSOME at 8400x magnification using an inverted microscope that was equipped with DIC/Nomarski differential interference contrast optics. A minimum of 200 motile spermatozoa per semen sample were evaluated, and the percentage of normal spermatozoa in each sample was determined.Results: Pregnancy occurred in 34 IUI cycles (CP rate per cycle: 21.8%, per patient: 30.6%). Based on the MSOME criteria, a significantly higher percentage of normal spermatozoa was found in the group of men in which the IUI cycles resulted in pregnancy (2.6+/-3.1%) compared to the group that did not achieve pregnancy (1.2+/-1.7%; P = 0.019). Logistic regression showed that the percentage of normal cells in the MSOME was a determining factor for the likelihood of clinical pregnancy (OR: 1.28; 95% CI: 1.08 to 1.51; P = 0.003). The ROC curve revealed an area under the curve of 0.63 and an optimum cut-off point of 2% of normal sperm morphology. At this cut-off threshold, using the percentage of normal sperm morphology by MSOME to predict pregnancy was 50% sensitive with a 40% positive predictive value and 79% specificity with an 85% negative predictive value. The efficacy of using the percentage of normal sperm morphology by MSOME in predicting pregnancy was 65%.Conclusions: The present findings support the use of high-magnification microscopy both for selecting spermatozoa and as a routine method for analysing semen before performing IUI.