[Comparison of two techniques for HPV genotyping in women with high-grade squamous intraepithelial lesion]. / Comparação entre duas técnicas de genotipagem do HPV em mulheres com lesão intra-epitelial de alto grau.

PURPOSE: The aim of this study was to compare the performance of two human papillomavirus (HPV) genotyping techniques, Linear Array and PapilloCheck, in women with high-grade squamous intraepithelial lesion (HSIL). METHODS: A total of 88 women with cytological diagnosis of HSIL were recruited at 2 reference centers in cervical pathology in Salvador, Bahia, Brazil, from July 2006 to January 2009. After the cytological diagnosis of HSIL, cervix cells were collected to determine the HPV genotype and a biopsy was obtained under colposcopic vision for histopathological analysis. After the confirmation of CIN2+ by histopathology, HPV genotyping was performed on 41 women by the Linear Array and PapilloCheck methods. RESULTS: Both tests showed an overall concordance rate for HPV detection of 97.2% (35/36). Of the 36 valid samples, 35 (97.2%) were positive in both tests and 1 (2.8%) was discordant, with the Linear Array indicating the presence of multiple types. The most prevalent HPV genotypes detected by the Linear Array technique were HPV 16, HPV 56, HPV 35, HPV 45, and HPV 70; and those detected by the PapilloCheck technique were HPV 16, HPV 56, HPV 11, HPV 35, and HPV 42. A similar rate of infection with multiple HPV types was observed with the two tests (72.5% with the Linear Array and 75.0% with the PapilloCheck). CONCLUSIONS: Linear Array genotyping assay and PapilloCheck showed equivalent performance for the detection of oncogenic HPV types in women with HSIL, with PapilloCheck having the advantage of being a method that avoids subjectivity when reading the HPV genotypes.

Comparação entre duas técnicas de genotipagem do HPV em mulheres com lesão intra-epitelial de alto grau / Comparison of two techniques for HPV genotyping in women with high-grade squamous intraepithelial lesion

OBJETIVO: Comparar o desempenho de duas técnicas de genotipagem de papilomavírus humano (HPV), Linear Array e PapilloCheck, em mulheres com lesão intra-epitelial de alto grau (LIAG). MÉTODOS: Foram selecionadas 88 mulheres com diagnóstico citopatológico de LIAG em 2 centros de referência em patologia cervical em Salvador, Bahia, no período de julho de 2006 a janeiro de 2009. Após o diagnóstico citopatológico de LIAG, foram realizadas a coleta de células do colo uterino para a genotipagem do HPV e a biópsia sob visão colposcópica para análise histopatológica do fragmento retirado. Posteriormente à confirmação de NIC2+ pelo exame histopatológico, foi realizada a genotipagem do HPV em 41 mulheres pelas técnicas Linear Array e PapilloCheck. RESULTADOS: Os dois testes apresentaram taxa de concordância geral para detecção do vírus HPV de 97,2% (35/36). Das 36 amostras válidas, 35 (97,2%) foram consideradas positivas em ambos os testes e apenas uma amostra (2,8%) foi considerada discordante. Os genótipos do HPV mais prevalentes detectados através da técnica do Linear Array foram: HPV 16, HPV 56, HPV 35, HPV 45 e HPV 70; e pela técnica PapilloCheck foram: HPV 16, HPV 56, HPV 11, HPV 35 e HPV 42. Foi observado índice semelhante de infecção por múltiplos tipos do HPV nos dois testes analisados (72,5% no Linear Array e 75,0% no PapilloCheck). CONCLUSÕES: Os testes de genotipagem Linear Array e PapilloCheck apresentaram um desempenho equivalente na detecção dos tipos de HPV oncogênicos em mulheres com LIAG, tendo o PapilloCheck a vantagem de ser um método que evita a subjetividade da leitura dos genótipos de HPV. .

PURPOSE: The aim of this study was to compare the performance of two human papillomavirus (HPV) genotyping techniques, Linear Array and PapilloCheck, in women with high-grade squamous intraepithelial lesion (HSIL). METHODS: A total of 88 women with cytological diagnosis of HSIL were recruited at 2 reference centers in cervical pathology in Salvador, Bahia, Brazil, from July 2006 to January 2009. After the cytological diagnosis of HSIL, cervix cells were collected to determine the HPV genotype and a biopsy was obtained under colposcopic vision for histopathological analysis. After the confirmation of CIN2+ by histopathology, HPV genotyping was performed on 41 women by the Linear Array and PapilloCheck methods. RESULTS: Both tests showed an overall concordance rate for HPV detection of 97.2% (35/36). Of the 36 valid samples, 35 (97.2%) were positive in both tests and 1 (2.8%) was discordant, with the Linear Array indicating the presence of multiple types. The most prevalent HPV genotypes detected by the Linear Array technique were HPV 16, HPV 56, HPV 35, HPV 45, and HPV 70; and those detected by the PapilloCheck technique were HPV 16, HPV 56, HPV 11, HPV 35, and HPV 42. A similar rate of infection with multiple HPV types was observed with the two tests (72.5% with the Linear Array and 75.0% with the PapilloCheck). CONCLUSIONS: Linear Array genotyping assay and PapilloCheck showed equivalent performance for the detection of oncogenic HPV types in women with HSIL, with PapilloCheck having the advantage of being a method that avoids subjectivity when reading the HPV genotypes. .

Prevalence of sexually transmitted infections among HIV-infected women in Brazil.

This study aimed to evaluate the prevalence of sexually transmitted infections (STIs) and associated risk factors in HIV-infected pregnant women followed for prenatal care in Salvador, Bahia. This was a cross-sectional study of 63 women seeking prenatal care at a reference center. Participants were interviewed regarding socio-epidemiological and clinical history, and were tested for HBsAg, anti-HCV, anti HTLV I/II, VDRL, Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma hominis, Ureaplasma urealyticum, CD4 count, and HIV plasma viral load. The main outcome variable was the presence of any STI. The mean age of patients was 28.2 years (16-40 years). 23 (36.5%) were diagnosed with at least one STI. The frequency of diagnoses was: HBV, 3.2%; HCV, 8.1%; HTLV I/II, 3.4%; syphilis, 9.5%; Chlamydia trachomatis, 11.1%; HPV, 15.0%; Mycoplasma hominis, 2.1%, and Ureaplasma urealyticum, 2.1%. No case of Neisseria gonorrhoeae was identified. No association was found between socio-epidemiological variables and the presence of an STI. CD4 T lymphocyte < 500 cells/µL (p=0.047) and plasma viral load >1,000 copies (p = 0.027) were associated with the presence of STI. STIs are frequent in pregnant women infected with HIV, and all HIV-infected pregnant women should be screened to decrease transmission of these pathogens and to protect their own health.

Comparative study of the expression of cellular cycle proteins in cervical intraepithelial lesions.

Interaction of human papilloma virus oncoproteins E6 and E7 with cell cycle proteins leads to disturbances of the cell cycle mechanism and subsequent alteration in the expression of some proteins, such as p16INK4a, cyclin D1, p53 and KI67. In this study, we compared alterations in the expression of these proteins during several stages of intraepithelial cervical carcinogenesis. Accordingly, an immunohistochemical study was performed on 50 cervical biopsies, including negative cases and intraepithelial neoplasias. The expression patterns of these markers were correlated with the histopathological diagnosis and infection with HPV. The p16INK4a, followed by Ki67, showed better correlation with cancer progression than p53 and cyclin D1, which recommends their use in the evaluation of cervical carcinogenesis. These monoclonal antibodies can be applied to cervical biopsy specimens to identify lesions transformed by oncogenic HPV, separating CIN 1 (p16INK4a positive) and identifying high-grade lesions by an increase in the cellular proliferation index (Ki67). In this way, we propose immunomarkers that can be applied in clinical practice to separate patients who need a conservative therapeutic approach from those who require a more aggressive treatment.

Padrão citológico de punção aspirativa do tecido tireoidiano morfologicamente normal / Cytologic pattern for fine-needle aspiration among morphologically normal thyroids

Nódulos tireoidianos pequenos com diagnóstico citológico de padrão folicular causam muita inquietação porque, se por um lado podem representar uma lesão que exige muito rigor no acompanhamento, por outro poderão representar material aspirado do tecido tireoidiano normal. OBJETIVO: Verificar o padrão citológico do tecido tireoidiano normal obtido através de punções aspirativa e não-aspirativa em cadáveres. MÉTODOS: Estudo observacional em que se procedeu a dissecção anatômica da tireóide normal e se efetuou a citoaspiração da glândula, em 38 casos. Dois patologistas cegos para a metodologia do estudo, sem conhecer a correspondência entre citologia e histologia, analisaram os esfregaços e os cortes histológicos. RESULTADOS: As tireóides normais forneceram diagnóstico citológico de bócio adenomatoso em 70,4 por cento das vezes para um observador e 92,6 por cento para o outro. Houve uma concordância regular entre os observadores, com kappa de 0,51 (p < 0,0001). Contrariamente ao que se esperava, nenhum caso exibiu padrão folicular nos preparados citológicos. CONCLUSAO: O aspecto citológico da tireóide normal em cadáveres foi semelhante ao do bócio adenomatoso.

P16(INK4a) expression as a potential prognostic marker in cervical pre-neoplastic and neoplastic lesions.

An immunohistochemical analysis with monoclonal antibody p16(INK4a) was performed in formalin-fixed, paraffin-embedded samples of 60 cases. The aim was to investigate in biopsies the expression of p16(INK4a) of normal uterine cervical tissue, pre-cancerous and cancerous lesions, and their relation with human papilloma virus (HPV) and HIV status. Three parameters were evaluated: percentage of p16(INK4a) positive cells, reaction intensity, and cell staining pattern. All of these parameters were statistically different when compared among different histological groups. However, logistic regression model showed that the reaction intensity was the best indicator of the expression of p16(INK4a). This expression increases from normal to invasive squamous carcinoma. Sixty-six percent of the patients with CIN grade 1 (CIN1) expressed p16(INK4a) (all these cases were infected with high risk HPV). Our study supports the hypothesis that p16(INK4a) expression in pre-cancerous lesions and cancers can be used to identify HPV-transformed cells. Of great interest for routine diagnostic use is the fact that immunohistochemical testing for p16(INK4a) seems to be capable of identifying HPV-positive cells and potentially recognizing those lesions with an increased risk of progression to high-grade lesions.

Prevalence of human Papilloma Virus in HIV-positive and HIV-negative patients in the State of Bahia: a pilot study

Human Papilloma Virus (HPV) plays a central role in the development of cervical cancer. However, other coexisting factors, such as HIV infection, must be present for this to occur. We evaluated the prevalence of HPV in HIV-positive and HIV-negative patients in the city of Salvador , Bahia, Brazil, and determined the most prevalent types of HPV in these patients. Fifty-five cases were selected from among patients attending three institutions providing cervical pathology services in the city of Salvador. HIV testing (Elisa/WB), HPV-DNA testing by PCR, colposcopy, cytology and biopsy were carried out in all patients. The histopathological results were classified as follows: 11 cases were normal/negative for neoplasia, 15 were diagnosed as cervical intraepithelial neoplasia grade 1 (CIN 1), 10 were CIN 2, 15 cases were CIN 3 and there were four cases of invasive squamous cell carcinoma. Among the 55 patients studied, 43 tested positive for HPV-DNA and 20 for HIV. All HIV-positive patients were positive for HPV-DNA. The most prevalent types of HPV were HPV 16, 52, 58, 53, 54, 33 and 51, and there was little difference between the groups of HIV-positive and HIV-negative patients with respect to the type of HPV encountered. The HIV-positive patients were found to be infected with a greater number of types of HPV than the HIV-negative patients. This study corroborates the existence of regional variations in the distribution of certain types of HPV, which is probably due to the particular ethnic constitution found in this region of Brazil.

Prevalence of human Papilloma Virus in HIV-positive and HIV-negative patients in the State of Bahia: a pilot study.

Human Papilloma Virus (HPV) plays a central role in the development of cervical cancer. However, other coexisting factors, such as HIV infection, must be present for this to occur. We evaluated the prevalence of HPV in HIV-positive and HIV-negative patients in the city of Salvador, Bahia, Brazil, and determined the most prevalent types of HPV in these patients. Fifty-five cases were selected from among patients attending three institutions providing cervical pathology services in the city of Salvador. HIV testing (Elisa/WB), HPV-DNA testing by PCR, colposcopy, cytology and biopsy were carried out in all patients. The histopathological results were classified as follows: 11 cases were normal/negative for neoplasia, 15 were diagnosed as cervical intraepithelial neoplasia grade 1 (CIN 1), 10 were CIN 2, 15 cases were CIN 3 and there were four cases of invasive squamous cell carcinoma. Among the 55 patients studied, 43 tested positive for HPV-DNA and 20 for HIV. All HIV-positive patients were positive for HPV-DNA. The most prevalent types of HPV were HPV 16, 52, 58, 53, 54, 33 and 51, and there was little difference between the groups of HIV-positive and HIV-negative patients with respect to the type of HPV encountered. The HIV-positive patients were found to be infected with a greater number of types of HPV than the HIV-negative patients. This study corroborates the existence of regional variations in the distribution of certain types of HPV, which is probably due to the particular ethnic constitution found in this region of Brazil.

Expressão de proteínas do ciclo celular e de moléculas de adesão na carcinogênese cervical / Expression of the protein cycle and adhesion molecules in the cervical carcinogenesis

Objetiva avaliar comparativamente a expressão das proteínas do ciclo celular p16INK4a, ciclina D1, p53 e Ki67 e das moléculas de adesão CD44v5 e CD44v6 nas diversas etapas da carcinogênese cervical correlacionando o padrão de expressão destas proteínas com a infecção pelo HPV. Casuística e métodos. Estudo imunoistoquímico com mAb p 16 INK4a, ciclina D1, p53, Ki67, CD44v5 e CD44v6 em 60 biópsias do colo uterino de pacientes com diagnóstico negativo para neoplasia (11), NIC1 (15), NIC2 (10), NIC3 (15) e carcinoma invasivo (9). Vinte foram HIV positivas e 46 foram HPV positivas. Avaliaram-se intensidades e padrões de expressão destes marcadores. Resultados. Existe correlação entre a expressão p16INK4a, Ki67, ciclina D1 e p53 e o diagnóstico histológico, sendo que os dois primeiros marcadores têm associação mais significante que a ciclina D1 e p53. Observamos redução da expressão de CD44v5 e CD44v6, particularmente nas NIC 3 e carcinoma e alteração da expressão destas moléculas durante a progressão da carcinogênse cervical. Não encontramos diferença na expressão de qualquer marcador nas amostras de pacientes HIV positivas e negativas em relação ao status da infecção pelo HPV. Após estudo da expressão destes marcadores, sugerimos um painel constituído de p16INK4a, Ki67 e CD44v6, visando prever a evolução das NICs. Conclusões. A reação positiva para Ki67 é importante indicador de lesão de alto grau, mas menos precisa para lesões de baixo grau. A p16INK4a discrimina os casos HPV negativos ou infectados pelo HPV de baixo risco, daqueles infectados pelo HPV de alto risco oncogênico em que já ocorreu transformação, permitindo identificar Lesões pré-cancerosas de alto grau ou câncer invasivo. A CD44v6 é util como marcador de risco de progressão em lesões intra-epiteliais. Este painel de anticorpos monoclonais poderá ser empregado para melhor caracterização dos exames histopatológicos cervicais. Além de ajudar a reduzir a subjetivade de interpretação dos patologistas, particularmente nas lesões de baixo grau.

Pulmonary involvement in patients with HTLV-I associated myelopathy

A frequência do comprometimento pulmonar em pacientes com doença neurológica associada ao primeiro vírus linfotrópico humano de células T(HTLV-I) tem sido demonstrado por vários autores na Africa, Asia e América Latina. Com o objetivo de estudar o envolvimento pulmonar em pacientes com mielopatia por HTLV-I (HAM) estudamos o lavado bronco-alveolar (LBA) de pacientes com HAM e 13 pacientes com mielopatias de outras etiologias. A contagem diferencial das células do LBA de pacientes com HAM demonstrou percentual de Linfócitos maior que 20 em 18 (82 por cento) dos pacientes enquanto aqueles com mielopatias de outra natureza mostraram linfocitose no LBA em apenas 2 (15 por cento). Concluímos que o pulmao se constitui em importante órgao para a patogênese de HAM.