Microbiome analysis of Brazilian women cervix reveals specific bacterial abundance correlation to RIG-like receptor gene expression.

The relationship among microbiome, immunity and cervical cancer has been targeted by several studies, yet many questions remain unanswered. We characterized herein the virome and bacteriome from cervical samples and correlated these findings with innate immunity gene expression in a Brazilian convenience sample of HPV-infected (HPV+) and uninfected (HPV-) women. For this purpose, innate immune gene expression data were correlated to metagenomic information. Correlation analysis showed that interferon (IFN) is able to differentially modulate pattern recognition receptors (PRRs) expression based on HPV status. Virome analysis indicated that HPV infection correlates to the presence of Anellovirus (AV) and seven complete HPV genomes were assembled. Bacteriome results unveiled that vaginal community state types (CST) distribution was independent of HPV or AV status, although bacterial phyla distribution differed between groups. Furthermore, TLR3 and IFNαR2 levels were higher in the Lactobacillus no iners-dominated mucosa and we detected correlations among RIG-like receptors (RLR) associated genes and abundance of specific anaerobic bacteria. Collectively, our data show an intriguing connection between HPV and AV infections that could foster cervical cancer development. Besides that, TLR3 and IFNαR2 seem to create a protective milieu in healthy cervical mucosa (L. no iners-dominated), and RLRs, known to recognize viral RNA, were correlated to anaerobic bacteria suggesting that they might be related to dysbiosis.

Preventing Uterine Cervix Cancer: The Clinical Meaning of Atypical Glandular Cells / Prevenção do câncer do colo uterino: Significado clínico das células glandulares atípicas

Abstract Objective To determine the prevalence of the atypical glandular cells (AGCs) cytology and to analyze its clinical significance in different age ranges. Methods Retrospective observational study using computerized data from the Brazilian National Cancer Institute, including women screened between January 2002 and December 2008. The women included were those with an AGC result who were properly followed-up with colposcopy and a second cytology. Results A total of 132,147 cytopathological exams were performed during the study period. Five-hundred and thirty-three (0.4%) women with AGC cytology were identified and, of these, 69.41% (370/533) were properly referred for colposcopy and a new cytology. Most of the women (79.2%) with a 1st or 2nd AGC cytology were between the ages of 25 and 54 years. The 2nd cytology demonstrated 67.6% (250/370) of normality, 24.5% (91/370) of squamous atypia, and 6.2% (23/370) of AGC, 0.8% (3/370) adenocarcinoma in situ and 0.8% (3/370) adenocarcinoma invasor. On biopsy of the women with a second AGC cytology, 43.4% (10/23) had normal histology, 43.4% (10/23) had squamous lesions, 8.7% (2/23) had invasive adenocarcinoma, and 1.2% (1/23) had an inconclusive report. All of the women with high-grade squamous intraepithelial lesion (HSIL) or invasive adenocarcinoma (respectively 5 and 2 patients), after a 2nd AGC cytology were 25 years old or older. Conclusion The prevalence of the AGC cytology was low in the studied population. Most of the AGC cytology cases occurred in adult women between the ages of 25 and 54. Although most of the patients had normal histology after follow-up, several of them presented with squamous intraepithelial lesions or invasive adenocarcinoma.

Resumo Objetivo Determinar a prevalência de citologia com laudo de células glandulares atípicas (AGCs, na sigla em inglês) e analisar a significância clínica nas diferentes faixas etárias Métodos Estudo observacional retrospectivo, usando os dados arquivados no sistema do Instituto Nacional de Câncer no Brasil, que incluiu mulheres rastreadas entre janeiro de 2002 a dezembro de 2008. As mulheres incluídas tinham citologia com resultado de AGCs, que foram acompanhadas com colposcopia e nova citologia Resultados Um total de132,147 exames citopatológicos foram incluídos durante o período de estudo. Quinhentas e trinta e três mulheres com citologia de AGC foram identificadas e destas, 69.41% (370) foram encaminhadas para colposcopia e nova citologia. A prevalência de citologia de AGC na população estudada foi 0.4%. A maioria das mulheres (79.22%) com resultado citológico de AGC tinham idade entre 25 e 54 anos. A segunda citologia demonstrou 67.56% (250/370) de normalidade, 24.5% (91/370) de atipias escamosas, e 6.2% (23/370) de AGC. Na biopsia das mulheres com a 2ª citologia de AGC, 43.4% (10/23) tinham histologia normal, 43.4% (10/23) tinha lesões escamosas, 8.7% (2/23) tinha adenocarcinoma invasor e 1.2% (1/23) tinha laudo inconclusivo. Todas as mulheres com lesões intraepiteliais escamosas de alto grau (HSIL, na sigla em inglês) ou adenocarcinoma invasor (respectivamente 5 e 2pacientes), após a 2ª citologia com AGC, tinham 25 anos de idade ou mais. Conclusão A prevalência de citologia com AGC foi baixa na população estudada. Muitos casos de citologia com AGC apareceram em mulheres adultas, entre 25 e 54 anos de idade. Embora a maioria das pacientes tiveram histologia normal após seguimento, várias apresentaram lesões intraepiteliais escamosas ou glandulares invasoras.

Preventing Uterine Cervix Cancer: The Clinical Meaning of Atypical Glandular Cells.

OBJECTIVE: To determine the prevalence of the atypical glandular cells (AGCs) cytology and to analyze its clinical significance in different age ranges. METHODS: Retrospective observational study using computerized data from the Brazilian National Cancer Institute, including women screened between January 2002 and December 2008. The women included were those with an AGC result who were properly followed-up with colposcopy and a second cytology. RESULTS: A total of 132,147 cytopathological exams were performed during the study period. Five-hundred and thirty-three (0.4%) women with AGC cytology were identified and, of these, 69.41% (370/533) were properly referred for colposcopy and a new cytology. Most of the women (79.2%) with a 1st or 2nd AGC cytology were between the ages of 25 and 54 years. The 2nd cytology demonstrated 67.6% (250/370) of normality, 24.5% (91/370) of squamous atypia, and 6.2% (23/370) of AGC, 0.8% (3/370) adenocarcinoma in situ and 0.8% (3/370) adenocarcinoma invasor. On biopsy of the women with a second AGC cytology, 43.4% (10/23) had normal histology, 43.4% (10/23) had squamous lesions, 8.7% (2/23) had invasive adenocarcinoma, and 1.2% (1/23) had an inconclusive report. All of the women with high-grade squamous intraepithelial lesion (HSIL) or invasive adenocarcinoma (respectively 5 and 2 patients), after a 2nd AGC cytology were 25 years old or older. CONCLUSION: The prevalence of the AGC cytology was low in the studied population. Most of the AGC cytology cases occurred in adult women between the ages of 25 and 54. Although most of the patients had normal histology after follow-up, several of them presented with squamous intraepithelial lesions or invasive adenocarcinoma.

OBJETIVO: Determinar a prevalência de citologia com laudo de células glandulares atípicas (AGCs, na sigla em inglês) e analisar a significância clínica nas diferentes faixas etárias MéTODOS: Estudo observacional retrospectivo, usando os dados arquivados no sistema do Instituto Nacional de Câncer no Brasil, que incluiu mulheres rastreadas entre janeiro de 2002 a dezembro de 2008. As mulheres incluídas tinham citologia com resultado de AGCs, que foram acompanhadas com colposcopia e nova citologia RESULTADOS: Um total de132,147 exames citopatológicos foram incluídos durante o período de estudo. Quinhentas e trinta e três mulheres com citologia de AGC foram identificadas e destas, 69.41% (370) foram encaminhadas para colposcopia e nova citologia. A prevalência de citologia de AGC na população estudada foi 0.4%. A maioria das mulheres (79.22%) com resultado citológico de AGC tinham idade entre 25 e 54 anos. A segunda citologia demonstrou 67.56% (250/370) de normalidade, 24.5% (91/370) de atipias escamosas, e 6.2% (23/370) de AGC. Na biopsia das mulheres com a 2ª citologia de AGC, 43.4% (10/23) tinham histologia normal, 43.4% (10/23) tinha lesões escamosas, 8.7% (2/23) tinha adenocarcinoma invasor e 1.2% (1/23) tinha laudo inconclusivo. Todas as mulheres com lesões intraepiteliais escamosas de alto grau (HSIL, na sigla em inglês) ou adenocarcinoma invasor (respectivamente 5 e 2 pacientes), após a 2ª citologia com AGC, tinham 25 anos de idade ou mais. CONCLUSãO: A prevalência de citologia com AGC foi baixa na população estudada. Muitos casos de citologia com AGC apareceram em mulheres adultas, entre 25 e 54 anos de idade. Embora a maioria das pacientes tiveram histologia normal após seguimento, várias apresentaram lesões intraepiteliais escamosas ou glandulares invasoras.

HPV Induces Changes in Innate Immune and Adhesion Molecule Markers in Cervical Mucosa With Potential Impact on HIV Infection.

While most HPV infections are asymptomatic and clear spontaneously, persistent infection with high-risk HPVs is associated with cervical cancer and with increased risk of HIV acquisition. Although several hypotheses have been proposed to explain this phenomenon, none has been confirmed. Our aim was to investigate the expression of host factors involved in the susceptibility to HIV infection among HPV-infected women. Cervical samples were collected to characterize the expression levels of HIV susceptibility markers in the mucosa of HPV-infected compared with HPV-uninfected women. No differences in the frequency of CCR5+, integrin α4ß7+, activated and memory CD4+ T-cell were detected between the groups. We additionally evaluated the expression levels of genes involved in innate immune responses and in cell adhesion. HPV infected patients expressed higher levels of TLR9 and lower levels of pattern recognition receptors that recognize RNA (TLR3, TLR7, and MDA5/IFIH1). We also detected an impaired IFN pathway, with an increased Type I IFN and a decreased IFNα2 receptor expression. HPV+ samples displayed reduced expression of genes for adherens and tight junctions. Taken together, these results suggest that although HPV infection does not result in the recruitment/activation of susceptible CD4+ T-cell in the female genital tract, it leads to changes in the innate antiviral immune responses and in cell adhesion that are likely to favor HIV infection.

Detection of sexually transmitted infections at a Brazilian gynecology center: high prevalence of co-infections / Detecção de infecções sexualmente transmissíveis em uma unidade ginecológica brasileira: alta prevalência de coinfecções

ABSTRACT Introduction: Sexually transmitted infections (STI) remain a major public health problem and surveillance is crucial for prevention and control strategies. Objective: Our aim was to assess the prevalence of STI in a reference center for gynecology in Rio de Janeiro, Brazil. Materials and methods: It is a cross-sectional study conducted between August 2016 and June 2017. Whole blood and cervical cells were collected from 62 women and tested for human papillomavirus (HPV), human immunodeficiency virus (HIV), syphilis, chlamydia, gonorrhea and herpes simples virus 1 and 2 (HSV-1/2). Cervical lesions were diagnosed by cytopathology and in some patients by colposcopy (79%). Other STI were evaluated during clinical examination. Results: Cervical lesions were detected by cytopathology examinations in 46.8% of patients; those with a history of four sexual partners were at higher risk of developing them. There was moderate agreement between the cytopathology e colposcopy results (Kappa = 0.69). The prevalence of HSV (96.7%), syphilis (6.4%) and HIV (3.2%) were higher than that described in the literature, while the prevalence of chlamydia (6.4%) and gonorrhea (1.6%) were similar. HPV was detected in 53.2% of women, 32.3% of which were infected by HPV 16. In the context of co-infections, 38 women (61.3%) presented more than one STI. Conclusion: Since most of the women analyzed were affected by more than one STI, our results suggest that routine screening for these infections at health centers would help in early detection, treatment and prevention of these infections. These measures would also impact on patients' cervical cancer control.

RESUMO Introdução: As infecções sexualmente transmissíveis (ISTs) ainda são um importante problema de saúde pública, e a vigilância é essencial para sua prevenção e seu controle. Objetivo: Avaliar a prevalência das ISTs em uma unidade de referência ginecológica no Rio de Janeiro, Brasil. Materiais e métodos: Trata-se de um estudo transversal realizado entre agosto de 2016 e junho de 2017. Foram coletados sangue total e células cervicais de 62 mulheres e realizados testes para detecção de papiloma vírus humano (HPV), vírus da imunodeficiência humana (HIV), sífilis, clamídia, gonorreia e vírus do herpes simples 1 e 2 (HSV-1/2). As lesões cervicais foram diagnosticadas por citopatologia, e em algumas pacientes, por colposcopia (79%). Outras ISTs foram avaliadas durante o exame clínico. Resultados: Foram encontradas lesões cervicais pelo exame citopatológico em 46,8% das pacientes; aquelas com história de quatro parceiros sexuais tiveram mais risco de desenvolvê-las. Houve concordância moderada entre os resultados da citopatologia e colposcopia (Kappa = 0,69). A prevalência de HSV (96,7%), sífilis (6,4%) e HIV (3,2%) foi maior que a descrita na literatura, enquanto a prevalência de clamídia (6,4%) e gonorreia (1,6%) foi similar. O HPV foi detectado em 53,2% das mulheres, sendo 32,3% delas infectadas pelo HPV 16. No contexto das coinfecções, 38 mulheres (61,3%) tinham mais de uma IST. Conclusão: Visto que a maioria das mulheres analisadas era acometida por mais de uma IST, nossos resultados sugerem que uma triagem rotineira dessas infecções nas unidades de saúde poderia auxiliar na detecção precoce, bem como no tratamento e na prevenção. Essas medidas também impactariam no controle do câncer cervical das pacientes.

Bonafide, type-specific human papillomavirus persistence among HIV-positive pregnant women: predictive value for cytological abnormalities, a longitudinal cohort study.

This study investigated the rate of human papillomavirus (HPV) persistence, associated risk factors, and predictors of cytological alteration outcomes in a cohort of human immunodeficiency virus-infected pregnant women over an 18-month period. HPV was typed through L1 gene sequencing in cervical smears collected during gestation and at 12 months after delivery. Outcomes were defined as nonpersistence (clearance of the HPV in the 2nd sample), re-infection (detection of different types of HPV in the 2 samples), and type-specific HPV persistence (the same HPV type found in both samples). An unfavourable cytological outcome was considered when the second exam showed progression to squamous intraepithelial lesion or high squamous intraepithelial lesion. Ninety patients were studied. HPV DNA persistence occurred in 50% of the cases composed of type-specific persistence (30%) or re-infection (20%). A low CD4+T-cell count at entry was a risk factor for type-specific, re-infection, or HPV DNA persistence. The odds ratio (OR) was almost three times higher in the type-specific group when compared with the re-infection group (OR = 2.8; 95% confidence interval: 0.43-22.79). Our findings show that bonafide (type-specific) HPV persistence is a stronger predictor for the development of cytological abnormalities, highlighting the need for HPV typing as opposed to HPV DNA testing in the clinical setting.

Bonafide, type-specific human papillomavirus persistence among HIV-positive pregnant women: predictive value for cytological abnormalities, a longitudinal cohort study

This study investigated the rate of human papillomavirus (HPV) persistence, associated risk factors, and predictors of cytological alteration outcomes in a cohort of human immunodeficiency virus-infected pregnant women over an 18-month period. HPV was typed through L1 gene sequencing in cervical smears collected during gestation and at 12 months after delivery. Outcomes were defined as nonpersistence (clearance of the HPV in the 2nd sample), re-infection (detection of different types of HPV in the 2 samples), and type-specific HPV persistence (the same HPV type found in both samples). An unfavourable cytological outcome was considered when the second exam showed progression to squamous intraepithelial lesion or high squamous intraepithelial lesion. Ninety patients were studied. HPV DNA persistence occurred in 50% of the cases composed of type-specific persistence (30%) or re-infection (20%). A low CD4+T-cell count at entry was a risk factor for type-specific, re-infection, or HPV DNA persistence. The odds ratio (OR) was almost three times higher in the type-specific group when compared with the re-infection group (OR = 2.8; 95% confidence interval: 0.43-22.79). Our findings show that bonafide (type-specific) HPV persistence is a stronger predictor for the development of cytological abnormalities, highlighting the need for HPV typing as opposed to HPV DNA testing in the clinical setting.

HPV DNA genotyping and methylation of gene p16 INK4A in cervical LSIL.

BACKGROUND: DNA methylation is the most important epigenetic change involved in the control of gene expression in human cells. Methylation of the p16(INK4a) gene occurs early in the development of cervical cancer. Low-grade squamous intraepithelial lesions (LSILs) are prevalent, and their behavior is variable. OBJECTIVE: To identify the HPV DNA type, detect the methylation status of the p16(INK4A) gene, and analyze their association with the cytological evolution of LSIL over a period of two years. METHODS: We conducted a cohort study with 40 participants. Cervical scrapings were collected for cytological and molecular analysis. HPV DNA detection and typing were performed by means of polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). Methylation-specific PCR was performed to detect methylation. RESULTS: HPV DNA was detected in 87% of the cases, and type 16 was the most frequent type. Methylation was detected in 11% of the cases and did not exhibit a significant correlation with the HPV type. Unfavorable cytological evolution exhibited a significant association with the presence of methylation. CONCLUSION: HPV 16 was the most frequently detected type of HPV in LSIL. Methylation of the p16(INK4A) gene was infrequent and occurred independent of the presence of HPV DNA. Methylation of the p16(INK4a) gene exhibited a significant correlation with persistence/progression of LSIL.

HIV/HPV co-infection during pregnancy in southeastern Brazil: prevalence, HPV types, cytological abnormalities and risk factors.

OBJECTIVE: HIV(+) pregnant women are at a higher risk of HPV infection and development of cervical cancer. Our objectives were to assess the prevalence and HPV types in HIV(+) pregnant women and to identify risk factors for HPV infection and cytological abnormalities. METHODS: Cervicovaginal smears were collected during pregnancy from 140 women. Partial HPV L1 gene and the exon 4 of the human TP53 gene (containing codon 72) were PCR-amplified and sequenced. Amplified products indicating multiple HPV infection were further cloned and sequenced. The association of demographic, obstetric and HIV-related clinical variables with HPV infection and cervical lesions was tested by univariate analyses, and significant factors were subsequently tested by logistic regression multivariate analysis. RESULTS: HPV DNA tested positive for 118 patients and HPV types were identified in 104 samples. Twenty-eight different types were found, HPV-16 and HPV-58 being the most prevalent. High-risk types were present in 79.8% of samples and multiple infections in 16.3%. Abnormal cervical smears were found in 44 patients (31.4%). Absolute CD4(+) T-cell counts below 350 were associated with HPV infection. Younger age was associated with cervical abnormalities and higher CD4(+) T-cell count was an apparent protective factor. CONCLUSIONS: We found a high prevalence of HPV infection and high-risk types in this cohort. Our results highlighted the relevance of immune system integrity rather than TP53 variants for protecting this highly vulnerable population to HPV infection and carcinogenesis.

Analysis of the use of fasciocutaneous flaps for immediate vulvar reconstruction.

OBJECTIVE: To analyze the use of immediate reconstruction techniques of the vulva after surgical resection, with fasciocutaneous flaps of the medial and/or posterior thigh. METHODS: We conducted a transversal, retrospective study to analyse the outcome of immediate surgical reconstruction with fasciocutaneous flaps in nine patients who underwent vulvectomy from May 2009 to August 2010. RESULTS: Mean age was 61 years (range 36-82). In 56% of cases, diagnosis was vulvar intraepithelial neoplasia (VIN), usual type. Radical vulvectomy was performed in 45% of patients, simple vulvectomy in 33% and wide resections in 22%. Eleven fasciocutaneous flaps were made, of which 36.3% were flap transpositions from the posterior thigh, 18.2% from the medial thigh, 18.2% were in advancement flaps, 18.2% simple advancement flaps and 9.1% flap rotation from the posterior thigh. There were no major losses of the flaps made. CONCLUSION: Thigh fasciocutaneous flaps are currently the best options for immediate reconstruction after resection of vulvar cancer due to the preservation of sensibility and tissue availability in the donor areas. The association of the Plastic Surgeon with the Gynecologist offers tranquility for patients and provides good postoperative results.

The R337H mutation in TP53 and breast cancer in Brazil.

BACKGROUND: Germline mutations in p53 are associated with the Li-Fraumeni Syndrome which is characterized by childhood cancers, including pediatric adrenal cortical carcinomas and early onset breast cancer. The high incidence of adrenal cortical carcinomas in southern Brazil is mostly attributed to the R337H mutation in TP53. The relatively high population frequency of this mutation in southern Brazil, along with the clustering of early onset breast cancer in Li-Frameni families, suggests this mutation may also be a low-penetrance breast cancer susceptibility polymorphism. METHODS: We undertook this study to evaluate the frequency of the R337H mutation in breast cancer patients from Rio de Janeiro, Brazil. R337H mutation status was determined in 390 unselected breast cases and 324 controls identified from clinics in Rio de Janeiro, Brazil using a PCR-based assay. RESULTS: Two of the breast cancer cases (0.5%) and none of the controls carried the mutation. Both cases had an early age at diagnosis (< 40 years old) and a family history of breast and other cancers. CONCLUSIONS: These data suggest genetic screening of young onset breast cancer patients should include testing for the R337H mutation.

Promoter hypermethylation of death-associated protein kinase and p16 genes in vulvar lichen sclerosus.

OBJECTIVE: The purpose of this study was to discuss our investigation of the hypermethylation of promoter regions of tumor suppressor genes, such as death-associated protein kinase (DAPK) and p16, in vulvar lichen sclerosus (LS), in comparison with a control group. MATERIALS AND METHODS: Promoter hypermethylation of DAPK and p16 was investigated using 24 vulvar biopsies of patients with LS who had received no previous treatment. The control group was composed of 15 patients with no vulvar disease. The DNA of subjects was treated with sodium bisulphate, and the genes under study were subjected to methylation-specific polymerase chain reaction. The resulting polymerase chain reaction products were amplified and analyzed using a 10% polyacrylamide gel. RESULTS: The mean age of the patients with LS was 57 years (the majority were postmenopausal). In the control group, the mean age of the patients was 50 years (p = .151). Methylation of the promoter region of DAPK was found in 4 (17%) of the 23 patients analyzed, and p16 promoter region methylation was found in 8 patients (35%). Two cases of methylation of the DAPK gene were also found to be methylated for the p16 gene. In the control group, no methylation was found in the patients analyzed for the DAPK gene and methylation was found in 3 (21%) of the 14 patients analyzed for the p16 gene (p = .190 and p = .316, respectively). CONCLUSIONS: Methylation of the DAPK and p16 genes, although not sufficient to dictate prognosis of the disease, should not be underestimated because it may form part of a process of genetic and epigenetic alterations that in the future could become relevant to malignant transformation.

Análise do emprego de retalhos fasciocutâneos para reconstrução vulvar imediata / Analysis of the use of fasciocutaneous flaps for immediate vulvar reconstruction

OBJETIVO: Analisar o emprego de técnicas de reconstrução imediata de vulva, pós-ressecção cirúrgica, com retalhos fasciocutâneos das faces medial e/ou posterior da coxa. MÉTODOS: Estudo de coorte transversal, retrospectivo, para análise do resultado da reconstrução cirúrgica imediata, com retalhos fasciocutâneos em nove pacientes submetidas à vulvectomia, no período de maio de 2009 a agosto de 2010. RESULTADOS: A média de idade foi 61 anos (variação 36 a 82 anos). Em 56% dos casos, o diagnóstico foi neoplasia intraepitelial vulvar (NIV) tipo usual. A vulvectomia radical foi realizada em 45% das pacientes, a vulvectomia simples em 33% e as ressecções amplas, em 22%. Foram confeccionados 11 retalhos fasciocutâneos, sendo 36,3% de transposições de retalho posterior de coxa, 18,2% de retalhos mediais de coxa, 18,2% de retalhos em avanço em V-Y, 18,2% de retalhos em avanço simples e 9,1% de rotação de retalho de região posterior de coxa. Não houve casos de perdas importantes dos retalhos confeccionados. CONCLUSÃO: Os retalhos fasciocutâneos de coxa são, atualmente, boas opções para a reconstrução imediata da vulva pós-ressecção oncológica devido à preservação da sensibilidade e da disponibilidade tecidual nas áreas doadoras. A associação do Cirurgião Plástico com o Ginecologista oferece tranquilidade às pacientes e determina bons resultados pós-operatórios.

OBJECTIVE: To analyze the use of immediate reconstruction techniques of the vulva after surgical resection, with fasciocutaneous flaps of the medial and/or posterior thigh. METHODS: We conducted a transversal, retrospective study to analyse the outcome of immediate surgical reconstruction with fasciocutaneous flaps in nine patients who underwent vulvectomy from May 2009 to August 2010. RESULTS: Mean age was 61 years (range 36-82). In 56% of cases, diagnosis was vulvar intraepithelial neoplasia (VIN), usual type. Radical vulvectomy was performed in 45% of patients, simple vulvectomy in 33% and wide resections in 22%. Eleven fasciocutaneous flaps were made, of which 36.3% were flap transpositions from the posterior thigh, 18.2% from the medial thigh, 18.2% were in advancement flaps, 18.2% simple advancement flaps and 9.1% flap rotation from the posterior thigh. There were no major losses of the flaps made. CONCLUSION: Thigh fasciocutaneous flaps are currently the best options for immediate reconstruction after resection of vulvar cancer due to the preservation of sensibility and tissue availability in the donor areas. The association of the Plastic Surgeon with the Gynecologist offers tranquility for patients and provides good postoperative results.

Microinvasive squamous carcinoma (FIGO stage IA1) of the cervix: are there colposcopic criteria for the diagnosis?

OBJECTIVE: The purpose of this study was to evaluate colposcopic sensitivity in the diagnosis of microinvasive squamous carcinoma of the cervix. STUDY DESIGN: We conducted a cross-sectional study in 151 patients from 1991-2008. The colposcopic findings of microinvasion suspicion were described by the International Federation for Cervical Pathology and Colposcopy in 2003. RESULTS: There has been colposcopic suspicion of invasion in 35 patients, which represents a sensitivity of 23%. The major colposcopic findings that were observed in the transformation zone included acetowhite epithelium in 21% (32/151 patients), coarse punctuation in 19% (29/151 patients), coarse mosaic in 17% (26/151 patients), and atypical vessels in 3.9% (6/151 patients). Suspicion of microinvasion was found in 14.5% of unsatisfactory colposcopy and in 8.6% of satisfactory colposcopy. CONCLUSION: The sensitivity of colposcopy in the diagnosis of microinvasive carcinoma of the cervix was low. Colposcopy plays an important role in directing the biopsy to the most suspicious area. The definitive diagnosis of microinvasive squamous carcinoma is established only by histologic study.

Série de casos de neoplasia intraepitelial vulvar tratados com imiquimode / Case series of vulvar intraepithelial neoplasia treated with imiquimod

O termo neoplasia intraepitelial vulvar (NIV) inclui as lesões precursoras do câncer de vulva, sendo divididas em NIV escamosa e não escamosa. A NIV escamosa compreende a NIV tipo usual, a qual se encontra relacionada à infecção pelo HPV, à multicentricidade de lesões, à atividade sexual e ao fumo,sendo mais frequentemente observada em mulheres jovens. Já a NIV diferenciada está relacionada ao líquen escleroso, não estando associada à infecção induzida por HPV, nem ao comportamento sexual, acometendo pacientes mais idosas. A NIV não escamosa está relacionada à doença de Paget e ao melanoma in situ. O tratamento padrão da NIV é cirúrgico e consiste na excisão ampla da lesão com margem de segurança. Este estudo objetiva avaliar ouso do imiquimode creme a 5% no tratamento da NIV tipo usual de forma isolada e/ou, associado ao procedimento cirúrgico. O imiquimode já é utilizadocom eficácia no tratamento de lesões condilomatosas genitais. Além disso, tem-se mostrado eficaz em lesões de NIV tipo usual.

The term vulvar intraepithelial neoplasia (VIN) includes the precursor lesions of vulvar cancer, being divided into squamous VIN and non-squamous VIN. The squamous VIN refers to the usual type VIN, which is related to HPV infection, multicentricity of lesions, sexual activity and smoking beingmore frequently observed in young woman. The VIN differentiated type, which is related to vulvar dermatoses, such as lichen sclerosus, is not associated with HPV-induced infection or sexual behavior and affects older patients. Thenon-squamous VIN is related to Paget's disease and melanoma in "situ".The standard VIN's treatment is a surgical wide excision of the lesion with a safety margin. This study aims to test the use of imiquimod 5% cream in thetreatment of usual type VIN isolated, and/or associated with surgical procedure. The imiquimod is already successfully used in the treatment of genital condylomatous lesions. Moreover, it has been shown to be effective in usual type VIN lesions.

Promoter hypermethylation patterns of death-associated protein kinase and p16 genes in vulvar lichen sclerosus.

OBJECTIVE: This article aimed to investigate the hypermethylation of promoter regions of tumor suppressor genes, such as death-associated protein kinase (DAPK) and p16, in vulvar lichen sclerosus (LS). MATERIALS AND METHODS: The promoter hypermethylation of DAPK and p16 was investigated from 15 vulvar biopsies of patients with LS who had had no previous treatment. DNA was treated with sodium bisulfate and underwent methylation-specific polymerase chain reaction of these genes. The amplified polymerase chain reaction products were analyzed by 10% polyacrylamide gel. RESULTS: The mean age of the patients was 57 years (most were postmenopausal). Methylation of the promoter region of DAPK was found in 2 (13%) of 15 patients analyzed, and p16 promoter region methylation was found in 7 patients (47%). The samples that showed DAPK methylation also showed p16 methylation. CONCLUSIONS: Methylation of DAPK and p16 represent alterations that might occur in cell cycle control in LS. The hypothesis is that patients who had methylated genes in this study, mainly the 2 cases in which there has been methylation in both studied genes, may be more susceptible to the development of differentiated vulvar intraepithelial neoplasia or vulvar cancer. Methylation may play a role in progress of vulvar carcinogenesis.

Epstein-Barr virus and human papillomavirus infection in vulvar lichen sclerosus.

OBJECTIVE: We investigated the presence of the Epstein-Barr virus (EBV) and human papillomavirus (HPV) in patients with vulvar lichen sclerosus (LS). MATERIALS AND METHODS: We investigated the presence of HPV and EBV from 34 vulvar biopsies of patients with LS who had had no previous treatment and from 17 normal vulvar brushings used as controls. We used polymerase chain reaction to amplify DNA sequences of these viruses. Human papillomavirus and EBV DNA detection was carried out using MY09/MY11 and TC67/TC69 consensus primers, respectively. The amplified polymerase chain reaction products were analyzed by 10% polyacrylamide gel. RESULTS: The mean age of the patients was 57 years old, with the majority postmenopausal. Human papillomavirus DNA was not found in the LS samples studied, but it was found in 23.2% (4/17) of the controls. However, EBV DNA was found in 26.5% (9/34) of the LS samples analyzed, and it was not found in the controls. CONCLUSIONS: Our results showed no relationship between HPV and LS. This result is in accordance with the literature. We have found 26.5% of EBV in our samples. This is a preliminary study, and the follow-up of these patients will elucidate whether EBV could play a role in cases of LS.

The presence of methylation of the p16INK4A gene and human papillomavirus in high-grade cervical squamous intraepithelial lesions.

Methylation is a chemical modification in which a methyl group (CH3) is added to the cytosine in the promoter region of the gene. It involves a very frequent epigenetic event that is found in many human cancers. Currently, there is no consensus on whether methylation of the p16 gene could be used as a biomarker in cervical intraepithelial neoplasia. The authors studied the presence of methylation of the p16 gene and human papillomavirus (HPV) DNA, and a possible relationship between them in high-grade squamous intraepithelial lesions of the cervix. This case-control study analyzed 27 high-grade squamous intraepithelial lesion samples and 20 normal cytology samples. To detect p16 methylation, methylation-specific polymerase chain reaction was used, and for HPV DNA detection the polymerase chain reaction was performed by using MY09/MY11 and GP5+/GP6+ consensus primers. The presence of methylation of the promoter region of the p16INK4a gene was detected in 55.6% of the samples from the case group, whereas it was detected only in 20% of the samples from the control group (P=0.005). HPV DNA was found in 66.7% of the samples from the case group, whereas only 15% from the control group (P=0.0001). The relationship between the presence of methylation of the p16 gene and HPV DNA did not prove statistically significant in the case group (P=0.67) or the control group (P=0.51). In conclusion, the presence of methylation of the p16 gene constituted an occurrence that was early but independent of the presence of HPV DNA.

Neoplasia intraepitelial cervical / Cervical intraepithelial neoplasia

A neoplasia intraepitelial cervical (NIC) representa uma das manifestações subclínicas da infecção pelo papilomavírus humano (HPV). O diagnóstico e o tratamento dessas lesões são de grande importância, pelo fato de estarem intimamente relacionadas à gênese do câncer do colo uterino. Este artigo propõe-se a fazer uma revisão sobre os principais aspectos da epidemiologia, da história natural, do diagnóstico e do tratamento das NIC.

Cervical intraepithelial neoplasia (CIN) represent one of subclinical manifestations by Human Papillomavirus (HPV) infection. Diagnostic and treatment of CIN are extremely important because they are related with the carcinogenesis of cervical cancer. This article attempts to carry out a review on the main epidemiological data, natural history, diagnostic and treatment of CIN.

Evaluation of DAPK gene methylation and HPV and EBV infection in cervical cells from patients with normal cytology and colposcopy.

OBJECTIVE: Evaluation of promoter methylation of the death-associated protein kinase (DAPK) gene and HPV and EBV infections in cervical cells from patients with normal cytology and colposcopy. STUDY DESIGN: Twenty women, who had been patients at the Institute of Gynecology of the Federal University of Rio de Janeiro (UFRJ) for routine examinations and who showed normal cytology and colposcopy, were selected for this work. Cervical brushings were used for DNA extraction, and the analysis of methylation patterns of the DAPK gene was done through chemical modification with sodium bisulfite. Analysis of viral infection was done using polymerase chain reaction (PCR). RESULTS: Of the 20 patients studied, six (30%) presented methylation of the DAPK gene, five (25%) presented infection with EBV and three (15%) presented coinfection with HPV/EBV. Associating methylation with viral infection, we found methylated DAPK in one patient (16%) with EBV, in two patients (33%) with co-infection and in three patients (50%) with no viral infection. CONCLUSIONS: In the present study, we verified, for the first time, the methylation pattern of the DAPK gene in cervical smears from patients with normal cytology and colposcopy. The results also showed the presence of viral infections in these patients. EBV infection, irrespective of whether associated with HPV or not, may contribute to cervical carcinogenesis as a cofactor. Methylation of the DAPK gene is associated with cell transformation, suggesting that DAPK methylation might be an important marker for the development of cervical epithelial neoplasias.