Absolute quantitative real-time polymerase chain reaction for the measurement of human papillomavirus E7 mRNA in cervical cytobrush specimens.

BACKGROUND: Few reports of the utilization of an accurate, cost-effective means for measuring HPV oncogene transcripts have been published. Several papers have reported the use of relative quantitation or more expensive Taqman methods. Here, we report a method of absolute quantitative real-time PCR utilizing SYBR-green fluorescence for the measurement of HPV E7 expression in cervical cytobrush specimens. RESULTS: The construction of a standard curve based on the serial dilution of an E7-containing plasmid was the key for being able to accurately compare measurements between cervical samples. The assay was highly reproducible with an overall coefficient of variation of 10.4%. CONCLUSION: The use of highly reproducible and accurate SYBR-based real-time polymerase chain reaction (PCR) assays instead of performing Taqman-type assays allows low-cost, high-throughput analysis of viral mRNA expression. The development of such assays will help in refining the current screening programs for HPV-related carcinomas.

Comparación del Cytobrush + Espátula de Ayre con el Papmolina para Papanicolaou, Hospital Vicente Corral Moscoso 2004

Estudio realizado con el objetivo de determinar la efectividad, expresada en criterios de calidad de la muestra y presencia de discariosis, del instrumento Papmolina frente al Cytobrush + Espátula de Ayre, utilizados para la toma de muestra de Papanicolaou. Se consideró universo a los pacientes que acudieron, en un período de nueve meses, a la consulta externa de Ginecología del HVCM de Cuenca para realizarse una prueba de Papanicolaou. El estudio se cumplió con 130 pacientes a quiénes se realizó 260 citologías. Se tomó dos muestras a cada paciente con un intervalo de un mes. Resultados: los resultados sobre la calidad de la muestra fueron una efectividad superior al 90con ambos instrumentos y la diferencia que le concede un porcentaje mayor al Papmolina (95,4vs 92,3) con el Cytobrush + Espátula de Ayre no fue estadísticamente significativa. Tampoco lo fue la diferencia entre las muestras satisfactorias con limitaciones obtenidas con el Cytobrush + Espïçatula de Ayre (7,7) y con el Papmolina (4,6). (p=0,301). No se obtuvo muestras satisfactorias. Conclusiones: el nuevo instrumento en comparación resulta tan efectivo como los ya existentes por sus elevados porcentajes de obtención de muestras adecuadas para realizar el diagnóstico citológico. Los hallazgos lo confirman

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.

Cytokeratin Profile in Exfoliative Cytology of Smokers

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

Evolução no diagnóstico citológico do câncer de colo uterino - Novas tecnologias para o rastreamento citológico cervical

The cancer of the uterine cervix remains a serious public health problem in developing countries with low social-economic conditions. Papanicolaou smear has been used to screen women for cervical cancer since 1940, and it is widely recognized as an effective method for preventing cervical cancer. In Brazil, conventional Papanicolaou is the most common method used in diagnosis cytopathological routine, involving the morphological cell analysis by microscopic examination of cervical samples smeared on glass slides. On the other hand, the liquid cytology is an advantageous method whose specimen collected is put into a fluid alcohol based to guarantee the potential use of residual cellular material in cytobrush. Moreover, this method permits additional diagnostic studies, preserving the morphological cell features as the conventional one does. The aim of the present study was report the historical evolution of different cervical cytologic tests, emphasizing the advantages and disadvantages between conventional Papanicolaou method and new technologies in liquid based cytology.

Exfoliative cytology of the oral mucosa: Comparison of two collection methods

This study compared the sampling efficacy cia cytobrush and metal spatula for exfoliative cytology of the oral mucosa. Thirty students with no detectable oral alterations upon clinical examination were submitted to exfoliative cytology of the lateral border of the tongue, using a metal spatula on the left side and a cytobrush on the right side. The smears were stained using the Papanicoiaou technique and evaluated for cellularity, cell type, cell distribution, homogeneity, and cellular distortion, as well as the presence ol mucus, inflammatory infiltrate, and hemorrhage. A statistical test (Z-test) with a 95% confdence interval (Cl) showed a significant difference between the metal spatula and cytobrush in terms of cellularity (p = 0.02) and homogeneity (p = 0.01). No difference between the two methods was observed regarding cell type (p = 0.4, Z-test) or cell distribution for the 95% confidence interval (p = 0.2, Fisher's test). Cell distortion and the presence of mucus were observed in five cases that used the metal spatula and in two cases that used the cytobrush. No hemorrhage or inflammatory infItrate was detected in any of the slides. Based on the results of this study, the cytobrush produced qualitatively better smears in terms of cellularity and homogeneity compared to the metal spatula.

Estudo citocolposcópico anorretal em pacientes com lesões cervicais intraepiteliais de alto grau

Pós-graduação em Ginecologia, Obstetrícia e Mastologia - FMB

Perfil de citocinas no soro e na secreção cervical de mulheres com lesão intraepitelial de baixo grau, lesão intraepitelial de alto grau e carcinoma epidermóide invasor

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Expressão da survivina em diferentes condições relacionadas à carcinogênese intra-bucal humana

Pós-graduação em Biopatologia Bucal - ICT

Avaliação da expressão de citoqueratinas na citologia esfoliativa da mucosa bucal de pacientes fumantes

The present study compared the expression of cytokeratins CK6, CK16 and CK19 and pan-cytokeratin (PAN) in oral mucosa cells between smokers and nonsmokers in order to determine the stage of cell differentiation and to consequently infer proliferative activity and expressions indicative of a potential for malignant differentiation. Thirty smokers and 30 non-smokers seen at the clinics of FOSJC-UNESP were screened. Smears were obtained from the left lateral border of the tongue with a cytobrush and slides were processed for immunohistochemistry using the antibodies reported Conventional microscopy was used for qualitative analysis. The results were analyzed statistically by the Z test, Fisher's exact test and comparison of two proportions (plus-4 confidence interval method). The expression of CK6 (p=0.002), CK16 (p=0.003), CK19 (p=0.0001) and PAN (p=0.008) was higher in oral mucosa smears from smokers compared to non- smokers. ln conclusion, increased epithelial proliferation is observed in the oral mucosa of smokers as demonstrated by the increased expression of CK6 and CKJ6, and these cells present alterations in epithelial maturation

Incidence of subclinical endometritis and its effects on reproductive performance of crossbred dairy cows

In dairy cattle, uterine infections are not life threatening and often unavoidable; however, they reduce fertility and increase the production costs of properties. The aim of this study was to evaluate the incidence of subclinical endometritis from 32 to 70 days in milk (DIM) and its effects on the reproductive performance of crossbred dairy cows. Lactating cows (Holstein/Gir; n = 172), with no history of retained placenta, without clinical signs of uterine infection were used. The body condition score (BCS) was evaluated on a scale from 1 to 5. Ultrasound examination was performed to evaluate uterine lining and ovarian activity, while vaginal mucus was analyzed by gloved hand. The diagnosis of subclinical endometritis was performed by endometrial cytobrush technique. The samples were collected, stained, and examined microscopically; positive cases for subclinical endometritis were considered with the presence of a parts per thousand yen5 % of neutrophils. Later, the cows were submitted to conventional artificial insemination or timed artificial insemination. The incidence of subclinical endometritis in the herd was 26 %, and this was not affected by the season of calving, presence of corpus luteum, DIM, and parity. Cows with a BCS a parts per thousand currency sign2.50 had a higher incidence of subclinical endometritis. The conception rate to first insemination and pregnancy rate at 150 days postpartum were not influenced by the presence of subclinical endometritis in crossbred dairy cows.

Prevalência de infecção clamidiana em mulheres com diagnóstico de infertilidade primária e secundária atendidas no ambulatório de esterilidade do Hospital das Clínicas da Faculdade de Medicna de Botucatu, Unesp

A infertilidade está se tornando um problema emergente de saúde pública em muitos países do mundo e, para muitos autores, esse aumento parece coincidir com o crescente papel desempenhado pela Chlamydia trachomatis. A infecção por C. trachomatis é uma das principais causas de lesão tubária que pode levar a oclusão desta ou processos aderentes que comprometem o complexo tubo-ovariano. O objetivo deste trabalho foi avaliar a prevalência de cervicite por Chlamydia trachomatis em mulheres diagnosticadas com infertilidade primária ou secundária atendidas no Ambulatório de Esterilidade do Hospital das Clínicas da Faculdade de Medicina de Botucatu, UNESP. Foram incluídas no estudo 112 mulheres atendidas no período de julho de 2008 a junho de 2009, que concordaram em participar do estudo e que responderam ao questionário para caracterização sóciodemográfica e ginecológica, sendo 62 pacientes com infertilidade primária e 50 com infertilidade secundária. Durante o exame especular, foi coletada secreção cervical com cytobrush para pesquisa de C. trachomatis pela técnica de reação em cadeia da polimerase (PCR) e conteúdo vaginal por meio de swab para avaliação da flora vaginal pelo método de Gram. Em relação à caracterização das pacientes, 66,7% eram moradoras de municípios com menos de 100.000 habitantes, 63,4% relataram ter vínculo empregatício, 98,2% declararam união estável, 74,1% eram brancas e 14,3% fumantes. A mediana de idade das pacientes no momento da inclusão no estudo foi de 28 anos (14-44). A mediana de idade à menarca e ao início da atividade sexual foi de 12 anos (9-17) e 16 anos (11-38), respectivamente. A mediana do tempo de infertilidade foi de 4 anos (1-17). Ainda nesse estudo, 67,9% das pacientes relataram mais de três relações sexuais por semana, 25,0% relataram infecção do trato genital inferior anteriormente ao estudo, 23,2% relataram... (Resumo Completo, clicar acesso eletrônico abaixo)

Immunocytochemistry associated with oral exfoliative cytology methodological analysis

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Prevalência de coinfecção e fatores associados ao Papilomavírus Humano (HPV) e à Chlamydia tracomatis em mulheres em idade reprodutiva atendidas em Program de Prevençao de Câncer do Colo Uterino na região sudeste do Brasil: sub-titulo / (se houver)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)