Quality of life in women with HPV and accurary of direct visual anal inspection with acetic acid and lugol compared to high-resolution anoscopy to diagnose intraepithelial anal lesion / Qualidade de vida em mulheres com HPV e a acurácia da inspeção visual anal direta com ácido acético e lugol comparada à anuscopia de alta resolução para diagnóstico da lesão intraepitelial anal

Abstract Objectives: to describe life quality of women with HPV and analyze the accuracy of anal visual inspection with acetic acid and lugol compared to high-resolution anoscopy in the detection of anal intraepithelial lesions. Methods: diagnostic evaluation study conducted with adult solid-organ transplant recipients and/or diagnosed with HIV and/or with a history of anogenital neoplasia, attended at the High Resolution Anoscopy outpatient clinic between April and December 2018. To calculate the accuracy parameters, 87 women participated: 44 with lesions and 43 without them. The 44 women with lesions were also subjected to a descriptive study to assess their life quality using the SF-36 questionnaire. Results: it was observed that almost half of the women in the study practiced vaginal, anal and oral intercourse without condom. History of HPV infection and anogenital neoplasia were the most prevailing with percentages of 81% and 72.7%, respectively. In five out of the eight domains of SF-36 they did not perform well in regard to life quality. Visual inspection with acetic acid and lugol presented sensitivity of 22.7% and specificity of 100%. Conclusion: unsatisfactory life quality was evidenced. Direct visual inspection with acetic acid and lugol should not be used in trials to spot intraepithelial HPV lesions, for it presents low sensitivity.

Resumo Objetivos: descrever a qualidade de vida das mulheres com HPVe analisar aacurácia da inspeção visual anal com ácido acético e lugol comparada à anuscopia de alta resolução para detecção de lesão intraepitelial anal. Métodos: estudo de avaliação diagnostica realizado com mulheres adultas transplantadas de órgãos sólidos e/ou diagnosticadas com HIV e/ou com antecedente de neoplasia anogenital, atendidas no ambulatório de AAR entre abril e dezembro de 2018. Para cálculo dos parâmetros de acurácia, participaram 87 mulheres: 44 com lesões e 43 sem lesões. Das 44 mulheres com lesão, também foi realizado um estudo descritivo para a avaliar a qualidade de vida utilizando-seo questionário SF-36. Resultados: Observou-se que quase metade das mulheres do estudo são adeptas ao intercurso vaginal, anal e oralesem preservativo. Os antecedentes de infecção pelo HPV e neoplasia anogenital foram os mais prevalentes com valores de 81% e 72, 7% respectivamente. Cinco, dos oito domínios do SF-36, não apresentaram boa performance em relação à qualidade de vida. A inspeção visual com ácido acético e lugol apresentou sensibilidade de 22,7%o e especificidade de 100,0%o. Conclusão: evidenciou-se qualidade de vida insatisfatória. A inspeção visual anal direta com ácido acético e lugol não deve ser usada na triagem de lesões intraepiteliais por HPV, pois apresenta baixa sensibilidade.

Profile of women with anal neoplasia associated with cervical neoplasia receiving care at a tertiary healthcare facility in northeastern Brazil / Perfil de mulheres com neoplasia anal associada à neoplasia cervical atendidas em um serviço de saúde terciário do nordeste do Brasil

Abstract Objective: To describe the epidemiological, clinical and laboratory profiles of women with anal neoplasia associated with cervical neoplasia attending a tertiary healthcare facility in northeastern Brazil. Methods: This epidemiological, descriptive study was conducted using a database from a cross-sectional study carried out between December 2008 and January 2016. Women with a diagnosis of cervical neoplasia associated with anal neoplasia were included in the present study. Results: Of the women with cervical neoplasia, 14% were found to have an anal intraepithelial lesion or anal cancer. Median age was 33 years, 68% were non-white, and 70% were from urban regions, had little schooling and low income. Most reported having had anoreceptive (73%) and unprotected intercourse (84%). Regarding symptoms, 7% reported bleeding and 11% pruritus. Overall, 10% of the sample tested positive for the human immunodeficiency virus. Anal cytology was abnormal in 92%. High-resolution anoscopy was abnormal in all cases. Histopathology revealed three cases of invasive carcinoma and high-grade lesions in 32% of the cases. Conclusion: Women with a diagnosis of anal and cervical neoplasia are often young, non-white women, who initiated their sexual life at an early age, were exposed to unprotected anoreceptive intercourse, live in urban centers, have little schooling and a low-income level.

Resumo Objetivo: Descrever os perfis epidemiológico, clínico e laboratorial de mulheres com neoplasia anal associada à neoplasia cervical atendidas em uma unidade de saúde terciária no nordeste do Brasil. Métodos: Este estudo epidemiológico e descritivo usou um banco de dados de um estudo transversal realizado entre dezembro de 2008 e janeiro de 2016. Mulheres com diagnóstico de neoplasia cervical associada à neoplasia anal foram incluídas no presente estudo. Resultados: Das mulheres com neoplasia cervical, 14% apresentaram lesão intra-epitelial anal ou câncer anal. A mediana de idade foi de 33 anos; 68% das pacientes não eram brancas e 70% eram provenientes de regiões urbanas, com baixa escolaridade e baixa renda. A maioria relatou histórico de relações sexuais anoreceptivas (73%) e desprotegidas (84%). Quanto aos sintomas, 7% relataram sangramento e 11% prurido. No geral, 10% das pacientes apresentaram serologia positiva para o vírus da imunodeficiência humana. A citologia anal foi anormal em 92% da amostra. A anuscopia de alta resolução foi anormal em todos os casos. A histopatologia revelou três casos de carcinoma invasivo e lesões de alto grau em 32% dos casos. Conclusão: As mulheres com diagnóstico de neoplasia anal e cervical geralmente são jovens, não brancas, que iniciaram sua vida sexual em idade precoce, foram expostas a relações sexuais anoreceptivas desprotegidas, moram em centros urbanos e têm baixa escolaridade e baixo nível de renda.

Human papillomavirus and risk factors for cervical adenocarcinoma in the state of Pernambuco, Brazil / Papilomavírus humano e fatores de risco para adenocarcinoma cervical no estado de Pernambuco, Brasil

Abstract Objectives: to determine the incidence of the main high oncogenic risk types of the human papillomavirus (HPV) ( 16, 18, 31 and 33) and the risk factors for cervical adenocarcinoma. Methods: a case-control study was carried out with 324 women (69 with adenocarcinoma and 260 healthy controls) between 2001 and 2014. Information on risk factors associated with adenocarcinomawere collected and the detection performed on HPVby using Polymerase Chain Reaction (PCR) method. Results: adenocarcinoma was associated with age ≥40 years old (OR=2.95; 95%CI=1.13-7.71), ≤3 years of schooling (OR=2.34; 95%CI=1.02-5.37), presence of HPV (OR=6.75; 95%CI=2.41-18.91),women in menopausal status (OR=4.76; 95%CI:1.70-13.31) black race (OR=6.71; 95%CI= 2.11-21.32) and never had undergone cervical cancer screening (OR=9.92; 95%CI:2.41-40.81). Andamong the HPV types detected, HPV 18 was observed to be strongly associated with adenocarcinoma of the cervix (OR=99.1; 95%CI=12.96-757.78). Conclusions: the factors associated with cervical adenocarcinoma were ≥40 years old, ≤3 years of schooling, black race, menopausal status, never had undergone cervical cancer screening and the presence of HPV.

Resumo Objetivos: determinar a incidência dos principais Papilomavirus Humano (HPV) de alto risco oncogênico (16, 18, 31 e 33) e os fatores associados ao adenocarcinoma do colo uterino. Métodos: realizado estudo de caso-controle com 324 mulheres (69 com adenocarcinoma e 260 controles saudáveis), de 2001 a 2014. Foram colhidas informações sobre fatores de risco associados ao adenocarcinoma e realizada a detecção do HPV pelo método da Reação em Cadeia da Polimerase (PCR). Resultados: o adenocarcinoma foi associado à idade >40 anos (OR=2,95; IC95%=1,13 - 7,71), escolaridade <3 anos (OR=2,34; IC95%=1,02 - 5,37), presença do HPV (OR=6,75; IC95%=2,41 - 18,91), mulher no estado menopausal (OR=4,76; IC 95%=1,70 - 13,31), raça negra (OR=6,71; IC95%=2,11 - 21,32) e nunca ter feito o exame de prevenção de Papanicolau (OR=9,92; IC95%=2,41 - 40,81). Entre os tipos de HPV encontrados observou-se que HPV 18 teve forte associação (OR=99,1; IC95%=12,96 - 757,78) com o adenocarcinoma de colo uterino. Conclusões: os fatores associados ao adenocarcinoma de colo uterino foram idade >40 anos, escolaridade <3 anos, raça negra, estado menopausal, nunca ter realizado o Papanicolau e presença do HPV.

Anal prevalence of HPV in women with pre-malignant lesion or cancer in the cervix and anal canal simultaneously: cross-sectional study.

OBJECTIVE: To determine the prevalence of anal HPV genotypes and associated factors in women with pre-malignant lesion or cancer in the cervix and anal canal. METHODS: A prospective, cross-sectional study analyzed DNA samples taken from women with cervical pre-malignant lesions or cancer to identify anal HPV by polymerase chain reaction (PCR). The prevalence of high-risk HPV (HR-HPV) in women with intraepithelial neoplasia and anal cancer was calculated; sociodemographic and clinical risk factors were identified using multivariate analysis. RESULTS: A total of 152 patients were included (mean age 37.8 ± 10.01 years), of whom 101 (66.4%) had anal HR-HPV. Fourteen different anal HPV types were identified. HPV 16 and 18 were found in 30 (52.6%) anal high-grade squamous intraepithelial lesions (HSIL), and HPV 31 and 33 in 21 (36.8%) lesions. In the logistic regression analysis, the factors that remained associated with HR-HPV types were: an anal histopathology report of HSIL or invasive carcinoma (odds ratio [OR] 8.96, 95% confidence interval [CI] 3.40-23.57; P<0.0001) and alcohol consumption (OR 2.20, 95% CI 1.01-4.80; P=0.04). CONCLUSION: Prevalence of HR-HPV is high in the anal canal of women with cervical and anal pre-malignant lesions simultaneously or cancer of the cervix and/or anal canal. HPV 16, 31, 33, and 18 were the four major genotypes identified.

Cervical determinants of anal HPV infection and high-grade anal lesions in women: a collaborative pooled analysis.

BACKGROUND: Cervical cancer screening might contribute to the prevention of anal cancer in women. We aimed to investigate if routine cervical cancer screening results-namely high-risk human papillomavirus (HPV) infection and cytohistopathology-predict anal HPV16 infection, anal high-grade squamous intraepithelial lesions (HSIL) and, hence, anal cancer. METHODS: We did a systematic review of MEDLINE, Embase, and the Cochrane library for studies of cervical determinants of anal HPV and HSIL published up to Aug 31, 2018. We centrally reanalysed individual-level data from 13 427 women with paired cervical and anal samples from 36 studies. We compared anal high-risk HPV prevalence by HIV status, cervical high-risk HPV, cervical cytohistopathology, age, and their combinations, using prevalence ratios (PR) and 95% CIs. Among 3255 women with anal cytohistopathology results, PRs were similarly calculated for all anal HSIL and HPV16-positive anal HSIL. FINDINGS: Cervical and anal HPV infections were highly correlated. In HIV-negative women, anal HPV16 prevalence was 41% (447/1097) in cervical HPV16-positive versus 2% (214/8663) in cervical HPV16-negative women (PR 16·5, 95% CI 14·2-19·2, p<0·0001); these values were 46% (125/273) versus 11% (272/2588) in HIV-positive women (4·4, 3·7-5·3, p<0·0001). Anal HPV16 was also associated with cervical cytohistopathology, with a prevalence of 44% [101/228] for cervical cancer in HIV-negative women (PR vs normal cytology 14·1, 11·1-17·9, p<0·0001). Anal HSIL was associated with cervical high-risk HPV, both in HIV-negative women (from 2% [11/527] in cervical high-risk HPV-negative women up to 24% [33/138] in cervical HPV16-positive women; PR 12·9, 95% CI 6·7-24·8, p<0·0001) and HIV-positive women (from 8% [84/1094] to 17% [31/186]; 2·3, 1·6-3·4, p<0·0001). Anal HSIL was also associated with cervical cytohistopathology, both in HIV-negative women (from 1% [5/498] in normal cytology up to 22% [59/273] in cervical HSIL; PR 23·1, 9·4-57·0, p<0·0001) and HIV-positive women (from 7% [105/1421] to 25% [25/101]; 3·6, 2·5-5·3, p<0·0001). Prevalence of HPV16-positive anal HSIL was 23-25% in cervical HPV16-positive women older than 45 years (5/20 in HIV-negative women, 12/52 in HIV-positive women). INTERPRETATION: HPV-based cervical cancer screening programmes might help to stratify anal cancer risk, irrespective of HIV status. For targeted secondary anal cancer prevention in high-risk groups, HIV-negative women with cervical HPV16, especially those older than 45 years, have a similar anal cancer risk profile to that of HIV-positive women. FUNDING: International Agency for Research on Cancer.

Evaluation of immunological parameters in pregnant women: low levels of B and NK cells / Avaliação dos parâmetros imunológicos em mulheres gestantes: baixos níveis de células B e NK

Abstract Objective To describe the immunological and hematological reference intervals of low-risk pregnant women. Methods A cross-sectional retrospective database analysis of a basic and translational study analyzing the hematological evaluation blood counts and immunophenotyping of TCD3 + , TCD4 + , TCD8 + , B, and natural killer (NK) cells of the peripheral blood in 79 low-risk pregnant women and of 30 control women from the state of Pernambuco, Brazil, was performed. Results No significant differences were detected between the hematological profiles of the 2nd and 3rd trimesters. Nevertheless, the median level of B cells decreased significantly in the 2nd (174 x 103 μL; p < 0.002) and 3rd trimesters (160 x 103 μL; p < 0.001), compared with the control group (296 x 103 μL). Similarly, the median level of NK cells was lower in the 2nd (134 x 103 μL; p < 0.0004) and 3rd trimesters (100 x 103 μL, p < 0.0004), compared with the control group (183 x 103 μL). In contrast, relative TCD4+ and TCD8+ levels increased in the 2nd and 3rd trimesters compared with the controls (TCD4 + : 2nd trimester = 59%; p < 0.001; 3rd trimester = 57%; p < 0.01; control = 50%; and TCD8 + : 2nd trimester = 31%; p < 0.001; 3rd trimester = 36%; p < 0.01; control = 24%). Conclusion Low-risk pregnant women have ~ 40% less B and NK cells in the peripheral blood, compared with non-pregnant women. These parameters may improve health assistance for mothers and contribute to define reference values for normal pregnancies.

Resumo Objetivo Descrever o intervalo de referência imunológico e hematológico de gestantes de baixo risco. Métodos Realizou-se uma análise retrospectiva, de um estudo básico e translacional, analisando o perfil hematológico e a imunofenotipagem das células TCD3 + , TCD4 + , TCD8 + , B e natural killer (NK) do sangue periférico de 79 gestantes de baixo risco e de 30 mulheres (controles) do estado de Pernambuco, Brasil. Resultados Não observamos diferenças significativas entre os perfis hematológicos do 2° e 3° trimestres. No entanto, houve redução das células B no 2° (média = 174 x 103 μL; p < 0,002) e no 3° trimestres (160 x 103 μL; p < 0,001), comparado como grupo controle (296 x 103 μL). A mediana das células NK foi menor no 2° (134 x 103 μL; p < 0,0004) e no 3° trimestres (100 x 103 μL; p < 0,0004), comparado com o grupo controle (183 x 103 μL). Porém, o percentual de TCD4+ e de TCD8+ aumentou no 2° e 3° trimestres em relação aos controles (TCD4 + : 2° trimestre = 59%; p < 0,001; 3° trimestre = 57%; p < 0,01; controle = 50%; e TCD8 + : 2° trimestre = 31%; p < 0,001; 3° trimestre = 36%; p < 0,01; controle = 24%). Conclusão Mulheres grávidas de baixo risco têm ~ 40% menos células B e NK no sangue periférico em comparação com mulheres não grávidas. Estes parâmetros podem melhorar a assistência à saúde das mães e contribuir para a definição de valores de referência para gestações normais.

Evaluation of Immunological Parameters in Pregnant Women: Low Levels of B and NK Cells.

OBJECTIVE: To describe the immunological and hematological reference intervals of low-risk pregnant women. METHODS: A cross-sectional retrospective database analysis of a basic and translational study analyzing the hematological evaluation blood counts and immunophenotyping of TCD3 + , TCD4 + , TCD8 + , B, and natural killer (NK) cells of the peripheral blood in 79 low-risk pregnant women and of 30 control women from the state of Pernambuco, Brazil, was performed. RESULTS: No significant differences were detected between the hematological profiles of the 2nd and 3rd trimesters. Nevertheless, the median level of B cells decreased significantly in the 2nd (174 × 103 µL; p < 0.002) and 3rd trimesters (160 × 103 µL; p < 0.001), compared with the control group (296 × 103 µL). Similarly, the median level of NK cells was lower in the 2nd (134 × 103 µL; p < 0.0004) and 3rd trimesters (100 × 103 µL, p < 0.0004), compared with the control group (183 × 103 µL). In contrast, relative TCD4+ and TCD8+ levels increased in the 2nd and 3rd trimesters compared with the controls (TCD4 + : 2nd trimester = 59%; p < 0.001; 3rd trimester = 57%; p < 0.01; control = 50%; and TCD8 + : 2nd trimester = 31%; p < 0.001; 3rd trimester = 36%; p < 0.01; control = 24%). CONCLUSION: Low-risk pregnant women have ∼ 40% less B and NK cells in the peripheral blood, compared with non-pregnant women. These parameters may improve health assistance for mothers and contribute to define reference values for normal pregnancies.

OBJETIVO: Descrever o intervalo de referência imunológico e hematológico de gestantes de baixo risco. MéTODOS: Realizou-se uma análise retrospectiva, de um estudo básico e translacional, analisando o perfil hematológico e a imunofenotipagem das células TCD3 + , TCD4 + , TCD8 + , B e natural killer (NK) do sangue periférico de 79 gestantes de baixo risco e de 30 mulheres (controles) do estado de Pernambuco, Brasil. RESULTADOS: Não observamos diferenças significativas entre os perfis hematológicos do 2° e 3° trimestres. No entanto, houve redução das células B no 2° (média = 174 × 103 µL; p < 0,002) e no 3° trimestres (160 × 103 µL; p < 0,001), comparado com o grupo controle (296 × 103 µL). A mediana das células NK foi menor no 2° (134 × 103 µL; p < 0,0004) e no 3° trimestres (100 × 103 µL; p < 0,0004), comparado com o grupo controle (183 × 103 µL). Porém, o percentual de TCD4+ e de TCD8+ aumentou no 2° e 3° trimestres em relação aos controles (TCD4 + : 2° trimestre = 59%; p < 0,001; 3° trimestre = 57%; p < 0,01; controle = 50%; e TCD8 + : 2° trimestre = 31%; p < 0,001; 3° trimestre = 36%; p < 0,01; controle = 24%). CONCLUSãO: Mulheres grávidas de baixo risco têm ∼ 40% menos células B e NK no sangue periférico em comparação com mulheres não grávidas. Estes parâmetros podem melhorar a assistência à saúde das mães e contribuir para a definição de valores de referência para gestações normais.

Cross-sectional study of anal intraepithelial lesions in women with cervical neoplasia without HIV.

OBJECTIVE: To evaluate the prevalence of anal intraepithelial lesions and associated risk factors in women with cervical neoplasia. METHODS: The present cross-sectional study enrolled patients with intraepithelial or invasive cervical neoplasia who had been referred to the lower genital tract pathology outpatient department of the Instituto de Medicina Integral Prof. Fernando Figueira, Recife, Brazil, between December 1, 2008, and December 31, 2009; patients with HIV infections were excluded. All participants underwent anal cytology and high-resolution anoscopy; sociodemographic and clinical risk factors were identified using multivariate analysis. RESULTS: There were 324 patients included and 37 (11.4%) had anal intraepithelial neoplasia. Factors associated with anal intraepithelial neoplasia in the multivariate analysis were being older than 35 years of age (P=0.002), having completed no more than 4 years of education (P=0.012), anomalous anal cytology (P=0.003), and anomalous high-resolution anoscopy findings (P<0.001); subclinical HPV lesions on vulvoscopy (P=0.057) were not associated with anal intraepithelial neoplasia. CONCLUSION: The prevalence of anal intraepithelial neoplasia was high among patients with cervical neoplasia who did not have HIV, particularly patients older than 35 years.

Validating competencies for an undergraduate training program in rural medicine using the Delphi technique.

INTRODUCTION: Worldwide, half the population lives in rural or remote areas; however, less than 25% of doctors work in such regions. Despite the continental dimensions of Brazil and its enormous cultural diversity, only some medical schools in this country offer students the opportunity to acquire work experience focused on medicine in rural or remote areas. The objective of the present study was to develop a framework of competencies for a longitudinal medical training program in rural medicine as an integrated part of medical training in Brazil. METHODS: Two rounds of a modified version of the Delphi technique were conducted. Initially, a structured questionnaire was elaborated, based on a literature review. This questionnaire was submitted to the opinion of 20 panelists affiliated with the Rural Medicine Working Party of the Brazilian Society of Family and Community Medicine. The panelists were asked to evaluate the relevance of the competencies using a five-point Likert-type scale. In this study, the consensus criterion for a competency to be included in the framework was it being deemed 'very important' or 'indispensable' by a simple majority of the participants, while the criterion for excluding a competency was that a simple majority of the panel members considered that it 'should not be included' or was 'of little importance'. When a consensus was not reached regarding a given competency, it was submitted to a second round to enable the panelists to re-evaluate the now dichotomized questions. RESULTS: Compliance in responding to the questionnaire was better among the panelists predominantly involved in teaching activities (85%; n=12) compared to those working principally in patient care (45%; n=8). The questionnaire consisted of 26 core competencies and 165 secondary competencies. After evaluation by the specialists, all the 26 core competencies were classified as relevant, with none being excluded and only eight secondary competencies failing to achieve a consensus. No new competencies were suggested. Of the competencies that failed to reach a consensus in the first round, seven were excluded from the framework in the second round, with most of these being associated with hospital procedures. CONCLUSIONS: A framework of competencies for a program in rural medicine was developed and validated. It consists of 26 core competencies and 158 secondary competencies that should be useful when constructing competency-based curricula in rural medicine for medical education in Brazil.

Influence of IL-6, IL-8, and TGF-β 1 gene polymorphisms on the risk of human papillomavirus-infection in women from Pernambuco, Brazil

Human papillomavirus (HPV) infections are strongly associated with the development of cervical intraepithelial neoplasias and invasive cervical cancer. Polymorphisms in cytokine-encoding genes and behavioural cofactors could play an important role in protecting an individual against viral infections and cancer. Here, we investigated whether IL-6 -174 G>C, IL-8 +396 G>T, and TGF-β1 +869 G>C and +915 G>C polymorphisms were associated with susceptibility to HPV infection in women from north-east (Pernambuco) Brazil. We analysed 108 healthy uninfected women (HC) and 108 HPV-positive women with cervical lesions. Genetic polymorphisms were assessed using Sanger sequencing and polymerase chain reaction-restriction fragment length polymorphism. Comparison of the distribution of the genotypic and allelic frequencies of the IL-18 +396 T>G polymorphism between HPV infected woman an uninfected controls showed that the GG genotype and G allele were both more frequent in the HC group, and were associated with protection from HPV infection (p = 0.0015; OR = 0.29 CI95% = 0.13-0.61; p = 0.0005; OR = 0.45 CI95% 0.29-0.7, respectively). Individuals from the control group could have previously had HPV infection that was spontaneously eliminated; however, it was undetectable at the time of sample collection. Based on our findings, we hypothesize that the IL-8 +396 G>T polymorphism could interfere with susceptibility to HPV infection, by modulating the ability of immune system to fight the virus.

Influence of IL-6, IL-8, and TGF-ß1 gene polymorphisms on the risk of human papillomavirus-infection in women from Pernambuco, Brazil.

Human papillomavirus (HPV) infections are strongly associated with the development of cervical intraepithelial neoplasias and invasive cervical cancer. Polymorphisms in cytokine-encoding genes and behavioural cofactors could play an important role in protecting an individual against viral infections and cancer. Here, we investigated whether IL-6 -174 G>C, IL-8 +396 G>T, and TGF-ß1 +869 G>C and +915 G>C polymorphisms were associated with susceptibility to HPV infection in women from north-east (Pernambuco) Brazil. We analysed 108 healthy uninfected women (HC) and 108 HPV-positive women with cervical lesions. Genetic polymorphisms were assessed using Sanger sequencing and polymerase chain reaction-restriction fragment length polymorphism. Comparison of the distribution of the genotypic and allelic frequencies of the IL-18 +396 T>G polymorphism between HPV infected woman an uninfected controls showed that the GG genotype and G allele were both more frequent in the HC group, and were associated with protection from HPV infection (p = 0.0015; OR = 0.29 CI95% = 0.13-0.61; p = 0.0005; OR = 0.45 CI95% 0.29-0.7, respectively). Individuals from the control group could have previously had HPV infection that was spontaneously eliminated; however, it was undetectable at the time of sample collection. Based on our findings, we hypothesize that the IL-8 +396 G>T polymorphism could interfere with susceptibility to HPV infection, by modulating the ability of immune system to fight the virus.

CCR2 and CCR5 genes polymorphisms in women with cervical lesions from Pernambuco, Northeast Region of Brazil: a case-control study.

Polymorphisms in chemokine receptors play an important role in the progression of cervical intraepithelial neoplasia (CIN) to cervical cancer (CC). Our study examined the association of CCR2-64I (rs1799864) andCCR5-Δ32 (rs333) polymorphisms with susceptibility to develop cervical lesion (CIN and CC) in a Brazilian population. The genotyping of 139 women with cervical lesions and 151 women without cervical lesions for the CCR2-64I and CCR5-Δ32 polymorphisms were performed using polymerase chain reaction-restriction fragment length polymorphism. The individuals carrying heterozygous or homozygous genotypes (GA+AA) for CCR2-64I polymorphisms seem to be at lower risk for cervical lesion [odds ratio (OR) = 0.37, p = 0.0008)]. The same was observed for the A allele (OR = 0.39, p = 0.0002), while no association was detected (p > 0.05) with CCR5-Δ32 polymorphism. Regarding the human papillomavirus (HPV) type, patients carrying the CCR2-64Ipolymorphism were protected against infection by HPV type 16 (OR = 0.35, p = 0.0184). In summary, our study showed a protective effect ofCCR2-64I rs1799864 polymorphism against the development of cervical lesions (CIN and CC) and in the susceptibility of HPV 16 infection.

CCR2 and CCR5 genes polymorphisms in women with cervical lesions from Pernambuco, Northeast Region of Brazil: a case-control study

Polymorphisms in chemokine receptors play an important role in the progression of cervical intraepithelial neoplasia (CIN) to cervical cancer (CC). Our study examined the association of CCR2-64I (rs1799864) andCCR5-Δ32 (rs333) polymorphisms with susceptibility to develop cervical lesion (CIN and CC) in a Brazilian population. The genotyping of 139 women with cervical lesions and 151 women without cervical lesions for the CCR2-64I and CCR5-Δ32 polymorphisms were performed using polymerase chain reaction-restriction fragment length polymorphism. The individuals carrying heterozygous or homozygous genotypes (GA+AA) for CCR2-64I polymorphisms seem to be at lower risk for cervical lesion [odds ratio (OR) = 0.37, p = 0.0008)]. The same was observed for the A allele (OR = 0.39, p = 0.0002), while no association was detected (p > 0.05) with CCR5-Δ32 polymorphism. Regarding the human papillomavirus (HPV) type, patients carrying the CCR2-64Ipolymorphism were protected against infection by HPV type 16 (OR = 0.35, p = 0.0184). In summary, our study showed a protective effect ofCCR2-64I rs1799864 polymorphism against the development of cervical lesions (CIN and CC) and in the susceptibility of HPV 16 infection.

Association between p21 Ser31Arg polymorphism and the development of cervical lesion in women infected with high risk HPV.

Infection by high-risk human papillomavirus (HR-HPV) and single nucleotide polymorphism (SNP) in genes involved in cell cycle control, as p21 and p27, are important factors in the development of different types of human cancers. This study aims at investigating whether both the p21 Ser31Arg and p27 V109G polymorphisms are associated with susceptibility to the development of cervical lesions in women HR-HPV positive. We analyzed 132 women HPV positive and with cervical lesions or CC and 154 healthy control (HPV negative and without cervical lesions). p21 Ser31Arg and p27 V109G polymorphisms were analyzed using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method and sequencing. The p21 31Arg allele was associated with susceptibility for the development of cervical lesions (P* = 0.0009), while p27 V109G polymorphism showed no significant differences for this association (P* = 0.89). However, the combined effect of the polymorphisms showed that the presence of the CC genotype (SNP p21 Ser31Arg) conferred protection for the development of cervical lesions (OR = 0.39). p21 Ser31Arg and p27 V109G polymorphisms were not associated with the grade of cervical lesions (CINI, CINII, and CINIII) or CC (P* > 0.05). The HR-HPV more frequent in this study were of 16 (57.6 %) and 18 (37.1 %) types; however, no association was observed when both polymorphisms and risk factors analyzed were compared (P* > 0.05). Our findings suggest a possible association between p21 Ser31tabArg polymorphism and susceptibility to the development of cervical lesions in women from Pernambuco. Brazil.

Tumor necrosis factor (TNF) alpha and interleukin (IL) 18 genes polymorphisms are correlated with susceptibility to HPV infection in patients with and without cervical intraepithelial lesion.

BACKGROUND: The Human Papillomavirus (HPV) predisposes 500 000 women to cervical cancer. Host genetic background may facilitate virus persistence in the uterine cervix. Polymorphisms in regulatory and coding regions of cytokine genes have been associated with susceptibility to some human diseases. AIM: This study aims at investigating whether TNFA -308 G/A and IL18 -137 G/C and -607 C/A polymorphisms are associated with susceptibility to HPV infection/progression to high-grade squamous intraepithelial lesion (HSIL). SUBJECTS AND METHODS: One hundred and twenty-two HPV infected and 132 HPV negative women (the latter used as healthy controls) were analysed. TNFA -308 G/A and IL18 (-137G/C and -607 C/A) polymorphisms were analysed using specific sequence polymorphism PCR (SSP-PCR). Univariate statistical analysis and a logistic regression were performed. RESULTS: The TNFA -308A allele was associated with susceptibility to HPV infection (p = 0.0008), while the IL18 -607A allele conferred protection against HPV infection (p = 0.0043). TNFA -308 G/A and IL18 (-137G/C and -607 C/A) polymorphisms were not associated with development of cervical lesions (p > 0.05). An association was also observed between smoking and susceptibility to the development of HSIL. CONCLUSION: The findings suggest an association between two TNFA SNPs and susceptibility to HPV infection in women from Northeast Brazil. The results need to be functionally validated and replicated in other populations with different ethnic backgrounds.

[Prevalence of HPV-induced lesions in the anal canal among women with cervical intraepithelial neoplasia 2 and 3: cross-sectional study]. / Prevalência da lesão HPV induzida em canal anal de mulheres com neoplasia intraepitelial cervical 2 e 3: um estudo de corte transversal.

PURPOSE: To determine the prevalence of HPV-induced lesions in the anal canal of women with cervical intraepithelial neoplasia (CIN) grade 2/3. METHODS: A cross-sectional study was carried out from December 2008 to June 2009, in Pernambuco, northeastern Brazil. Only women with grade 2/3 CIN were included, and those who could not undergo anoscopy during their first visit were excluded. A cyttobrush was used for sample collection in order to identify HPV DNA through PCR and anal cytology. An anal biopsy was obtained in cases of abnormal anal cytology or major alterations in high resolution anoscopy (HRA). RESULTS: Thirty-two percent (n=37/115) of HRA were normal and 63.5% (n=73/115) showed acetowhite epithelium. Twenty-two percent (n=26/115) of anal cytologies were abnormal. Among the latter, 12.2% (n=14/26) were low-grade anal intraepithelial lesions and 3.4% (n=4/26) were high-grade anal intraepithelial lesions. Twenty-two anal biopsies were performed, 13.7% of which (n=3/22) were grade 2 anal intraepithelial neoplasia (AIN2) and 9% (n=2/22) were grade 3 AIN. Th HPV DNA was identified in 72.1% of cases (n=83/115). CONCLUSION: Women with CIN grade 2/3 showed a high prevalence of anal HPV infection and HPV-induced lesions.

Prevalência da lesão HPV induzida em canal anal de mulheres com neoplasia intraepitelial cervical 2 e 3: um estudo de corte transversal / Prevalence of HPV-induced lesions in the anal canal among women with cervical intraepithelial neoplasia 2 and 3: cross-sectional study

OBJETIVO: Determinar a prevalência da lesão anal induzida por HPV em mulheres com neoplasia intraepitelial cervical grau 2/3 (NIC2/3).MÉTODOS: Estudo transversal, realizado no período de dezembro de 2008 a junho de 2009, no Estado de Pernambuco, nordeste do Brasil. Foram incluídas no estudo apenas mulheres com diagnóstico de NIC2/3 confirmado por biópsia e excluídas aquelas que não realizaram exame na primeira visita. As amostras para identificação do DNA de HPV anal por PCR e citologia anal foram coletadas com escovinha endocervical. A biópsia anal foi realizada nos casos de citologia anal anormal ou alterações maiores na anuscopia de alta resolução (AAR).RESULTADOS: Das AARs, 32,1% (n=37/115) foram normais e 63,5% (n=73/115) exibiram epitélio acetobranco. Vinte e dois por cento das citologias anais (n=26/115) foram anormais. Dentre elas, 12,2% (14/26) corresponderam à lesão intraepitelial anal de baixo grau e 3,4% (n=4/26), a lesão intraepitelial anal de alto grau. Foram realizadas 22 biópsias, das quais 13,7% (n=3/22) tiveram diagnóstico de neoplasia intraepitelial anal (NIA2) e 9% (n=2/22), NIA 3. Identificou-se 72,1% (n=83/115) de DNA do HPV nas amostras.CONCLUSÃO: Mulheres com NIC2/3 apresentam elevada prevalência de infecção por HPV e lesão HPV induzida em canal anal.

PURPOSE: To determine the prevalence of HPV-induced lesions in the anal canal of women with cervical intraepithelial neoplasia (CIN) grade 2/3.METHODS: A cross-sectional study was carried out from December 2008 to June 2009, in Pernambuco, northeastern Brazil. Only women with grade 2/3 CIN were included, and those who could not undergo anoscopy during their first visit were excluded. A cyttobrush was used for sample collection in order to identify HPV DNA through PCR and anal cytology. An anal biopsy was obtained in cases of abnormal anal cytology or major alterations in high resolution anoscopy (HRA).RESULTS: Thirty-two percent (n=37/115) of HRA were normal and 63.5% (n=73/115) showed acetowhite epithelium. Twenty-two percent (n=26/115) of anal cytologies were abnormal. Among the latter, 12.2% (n=14/26) were low-grade anal intraepithelial lesions and 3.4% (n=4/26) were high-grade anal intraepithelial lesions. Twenty-two anal biopsies were performed, 13.7% of which (n=3/22) were grade 2 anal intraepithelial neoplasia (AIN2) and 9% (n=2/22) were grade 3 AIN. Th HPV DNA was identified in 72.1% of cases (n=83/115).CONCLUSION: Women with CIN grade 2/3 showed a high prevalence of anal HPV infection and HPV-induced lesions.

Anal cytology in women with cervical intraepithelial or invasive cancer: interobserver agreement / Citologia anal em mulheres com neoplasia intraepitelial ou invasiva cervical: concordância interobservadores

ABSTRACTIntroduction:Incidence rates of anal cancer have been rising worldwide in the last 20 years. Due to embryological, histological and immunohistochemical similarities between the anal canal and the cervix, routine screening with anal cytology for precursor lesions in high-risk groups has been adopted. Objective: To determine interobserver agreement for the diagnosis of anal neoplasia by anal cytology.Material and methods:A cross-sectional observational study was conducted in 324 women with cervical intraepithelial or invasive cancers, for screening of anal cancer, from December 2008 to June 2009. Three hundred twenty-four cytological samples were analyzed by three cytopathologists. Cytological evaluation was based on the revised Bethesda terminology; samples were also classified into negative and positive for atypical cells. We calculated the kappa statistic with 95% confidence interval (95% CI) to assess agreement among the three cytopathologists.Results:Interobserver agreement in the five categories of the Bethesda terminology was moderate (kappa for multiple raters: 0.6). Agreement among cytopathologists 1, 2 and 3 with a consensus diagnosis was strong (kappa: 0.71, 0.85 and 0.82, respectively).Conclusion:Interobserver agreement in anal cytology was moderate to strong, indicating that cervical cytomorphological criteria are reproducible also in anal samples.

RESUMOIntrodução:O número de casos de câncer de canal anal vem aumentando nos últimos 20 anos no mundo. Devido às similaridades embriológicas, histológicas e imuno-histoquímicas do canal anal com o colo uterino, adotou-se a citologia anal para rastreamento das lesões precursoras desse tipo de câncer em grupos de risco.Objetivo:Determinar a concordância interobservadores na citologia anal e a concordância entre os diagnósticos citológico e histopatológico no rastreamento das neoplasias anais.Material e métodos:Foi realizado um estudo observacional do tipo corte transversal para rastreamento de câncer anal em 324 mulheres com neoplasias intraepiteliais ou invasivas cervicais, no período de dezembro de 2008 a junho de 2009. Foram colhidas amostras citológicas anais, as quais foram analisadas por três citopatologistas; a seguir, elas foram classificadas de acordo com o consenso Bethesda 2001, sendo agrupadas em negativas e positivas para células atípicas. Biópsias e reação em cadeia de polimerase (PCR) para papilomavírus humano (HPV) foram realizadas para verificar a concordância interobservadores. Foi aplicado o coeficiente kappa múltiplo e simples, bem como o seu intervalo de confiança de 95%.Resultados:A concordância interobservadores, incluindo todas as categorias diagnósticas, foi moderada (coeficiente kappa múltiplo: 0,6). A concordância para identificar citologias anormais entre os citopatologistas 1, 2 e 3 com o diagnóstico de consenso foi forte (coeficiente de kappa simples: 0,71; 0,85 e 0,82; respectivamente).Conclusão:A concordância interobservadores na citologia anal foi de moderada a forte, indicando que os critérios citomorfológicos são reprodutíveis na interpretação de material anal.

High-resolution anoscopy in women with cervical neoplasia.

OBJECTIVE: To describe high-resolution anoscopy (HRA) findings and compare them with histopathology results. METHODS: In a cross-sectional, observational study performed between December 2008 and December 2009, women receiving care at a center in Recife, Brazil, after a histopathologic diagnosis of cervical intraepithelial neoplasia or cervical cancer were screened for anal neoplasia by HRA. Patients with anal lesions were divided into groups A (metaplasia and/or human papillomavirus infection) and B (anal intraepithelial neoplasia [AIN]). Patients with squamous cell atypia in group A and all patients in group B underwent histopathologic analysis. Agreement between HRA and histopathology findings was estimated for group B. RESULTS: HRA was done in 324 women, 204 (63.0%) of whom had anal lesions. Overall, 169 cases (82.8%) were classified as group A and 35 (17.2%) as group B. Histopathologic data were obtained for 28 of the 35 group B cases. Histopathology was suggestive of AIN in 19 (67.9%), resulting in a κ coefficient of 0.45 (95% confidence interval [CI] 0.26-0.65; P<0.001). Relative to histopathology, HRA had sensitivity of 57.6% (95% CI 40.8%-72.7%), specificity of 86.1% (95% CI 75.7%-92.5%), positive likelihood ratio of 4.1 (95% CI 3.1-5.5), negative likelihood ratio of 0.5 (95% CI 0.4-0.5), and accuracy of 76.5% (95% CI 67.2%-83.8%). CONCLUSION: HRA findings can be systematized, reducing the subjectivity of interpretation.

Quantifying mRNA and microRNA with qPCR in cervical carcinogenesis: a validation of reference genes to ensure accurate data.

A number of recent studies have catalogued global gene expression patterns in a panel of normal, tumoral cervical tissues so that potential biomarkers can be identified. The qPCR has been one of the most widely used technologies for detecting these potential biomarkers. However, few studies have investigated a correct strategy for the normalization of data in qPCR assays for cervical tissues. The aim of this study was to validate reference genes in cervical tissues to ensure accurate quantification of mRNA and miRNA levels in cervical carcinogenesis. For this purpose, some issues for obtaining reliable qPCR data were evaluated such as the following: geNorm analysis with a set of samples which meet all of the cervical tissue conditions (Normal + CIN1 + CIN2 + CIN3 + Cancer); the use of individual Ct values versus pooled Ct values; and the use of a single (or multiple) reference genes to quantify mRNA and miRNA expression levels. Two different data sets were put on the geNorm to assess the expression stability of the candidate reference genes: the first dataset comprised the quantities of the individual Ct values; and the second dataset comprised the quantities of the pooled Ct values. Moreover, in this study, all the candidate reference genes were analyzed as a single "normalizer". The normalization strategies were assessed by measuring p16INK4a and miR-203 transcripts in qPCR assays. We found that the use of pooled Ct values, can lead to a misinterpretation of the results, which suggests that the maintenance of inter-individual variability is a key factor in ensuring the reliability of the qPCR data. In addition, it should be stressed that a proper validation of the suitability of the reference genes is required for each experimental setting, since the indiscriminate use of a reference gene can also lead to discrepant results.