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.

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.

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.

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.

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.

Prevalência dos genótipos do papilomavírus humano: comparação entre três métodos de detecção em pacientes de Pernambuco, Brasil / Prevalence of human papillomavirus genotypes: comparison between three detection methods in patients of Pernambuco, Brazil

OBJETIVO: comparar três métodos para detecção do HPV e determinar a prevalência dos genótipos encontrados. MÉTODOS: um total de 120 amostras de raspagem da região cervical de mulheres portadoras de neoplasia intraepitelial cervical foram analisadas pela reação em cadeia da polimerase convencional, usando os sistemas de primers MY09/11, GP05+/06+ e pela Nested-PCR. As amostras foram submetidas à extração de DNA e, logo após, amplificadas com os primers GH20 e PC04 (β-globina) para verificação da qualidade do DNA obtido e pela reação em cadeia da polimerase convencional e Nested-PCR. Os fragmentos amplificados foram visualizados em gel de agarose a 1,2 por cento, corados com Blue Green Loading Dye I. As amostras positivas foram sequenciadas usando o sequenciador automático de DNA "MegaBACE 1000". Para análise estatística foram utilizados os teste do Χ2 e o de Fisher com nível de significância de 5 por cento. RESULTADOS: quinze amostras não se amplificaram para os primers de β-globina, sendo eliminadas do estudo. Das amostras restantes, 40 por cento (42/105) foram positivas para os primers MY09/11, 98 por cento (103/105) para os primers GP05+/06+ e 92 por cento (97/105) para Nested-PCR. Considerado as técnicas MY09/11 e GP05+/06+, foi possível observar 100 por cento de amostras positivas para o HPV. Neste estudo, a prevalência dos genótipos foi de 58, 23, 5, 4 e 3 por cento para HPVs 16, 18, 31, 33 e 56, respectivamente. Os HPV 67 e 83 apresentaram 2 por cento e os HPV 6, 11, 58 e candHPV85, 1 por cento cada. A prevalência dos genótipos neste estudo está de acordo com o reportado em todo o mundo (IC95 por cento=0,4657-0,8976). CONCLUSÕES: para obter resultados mais confiáveis, é necessário o uso de mais que um sistema de primers para detecção do HPV. Acredita-se que as três técnicas estudadas são importantes e adequadas para o diagnóstico clínico do HPV quando apropriadamente combinadas.

PURPOSE: to compare three methods for the detection of HPV infection and to determine the prevalence of the genotypes found. METHODS: a total of 120 cervical scrape samples from patients with cervical intraepithelial neoplasia were analyzed by the conventional polymerase chain reaction using the MY09/11 and GP05+/06+ primers, and by the Nested polymerase chain reaction. The samples were subjected to DNA amplification with the GH20 and PC04 primers (β-globin) to verify DNA quality and also by polymerase chain reaction and Nested polymerase chain reaction. The amplicons were visualized in 1.2 percent agarose gel stained with Blue Green Loading Dye I. Positive samples also were sequenced using the automatic DNA sequencer "MegaBACE 1000". The Χ2 and Fisher tests were used for statistical analysis with the level of significance set at 5 percent. RESULTS: fifteen samples were eliminated from the study because they failed to amplify the β-globin gene. Of the remaining samples, 40 percent (42/105) were positive using primers MY09/11, 98 percent (103/105) using primers GP05+/06+, and 92 percent (97/105) using Nested-PCR. With the MY09/11 and GP05+/06+ techniques, it was possible to obtain 100 percent HPV-positive samples. In this study, the prevalence of the genotypes found was 57, 23, 5, 4 and 3 percent for HPV genotypes 16, 18, 31, 33 and 56, respectively. HPVs 67 and 83 were present in 2 percent, and genotypes 6, 11, 58 and candHPV85 were present in 1 percent each. The prevalence of the more common genotypes (HPV 16 and 18) in this study agrees with that reported worldwide (IC95 percent=0.4657-0.8976). CONCLUSIONS: to obtain more reliable results, it is necessary the use of more than one primer system to detect HPV infections. We believe that the three techniques studied are important and suitable for the clinical diagnosis of HPV, when they are appropriately combined.