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Introduction: Human Papillomavirus (HPV) infections are of significant concern in men, given its potential impact on their health and the risk of transmission to partners. Understanding and addressing this infection in men is crucial to evaluate the effectiveness of vaccination in reducing HPV-related diseases. Objective: To assess the impact of HPV vaccination, potential genotype shifts, and adverse effects, through a prospective study conducted with male university students. Methods: The study involved 286 volunteers who were examined at Sexually Transmitted Disease Clinics at the Universidade Federal Fluminense in Niterói, Rio de Janeiro, Brazil. The HPV prevalence was evaluated using generic PCR, genotyped by DNA microarray and monitored adverse effects. Results: The findings of this study revealed the absence of moderate or severe adverse effects. Genetic shifts were observed, including the disappearance of oncogenic HPV types 16 and 18. Surprisingly, even after completing the full vaccine regimen, students still harbored HPV11 in the oral tract. Furthermore, persistent HPV 6 and 11 infections were identified in three students, who had pre-existing infections prior to vaccination, at the follow-up visit. Multivariate analysis uncovered independent associations, notably an increased risk of HPV infection in the oral tract among men who have sex with men. HPV prevalence rates remained low both before and after the vaccination scheme (T0: 14.7%, T1: 8.7%). Even after the full vaccination scheme, the prevalence remained similar at T2 (14.6%), with no statistically significant differences recorded. HPV11 emerged as the most prevalent type throughout the study, followed by HPV6. Vaccine genotypes were detected in a significant proportion of samples at T0 (85.4%), T1 (89.5%), and T2 (100%). Conclusion: Overall, this study suggests that vaccination may represent a promising approach to reducing HPV-related health risks. These findings shed light on the potential benefits and challenges of HPV vaccination, emphasizing the need for continued monitoring and vaccination efforts.

Introdução: As infecções causadas pelos papilomavírus humanos (HPV) são responsáveis pelo desenvolvimento de cânceres em diversos sítios anatômicos humanos. Entretanto, a história natural da infecção em sítios que não a cérvice uterina não é muito clara. Objetivo: Avaliar infecções orais, genitais e anais por HPV, correlacionando taxas de prevalência do vírus e seus genótipos aos sítios de infecção e a fatores de risco sócio-demográficos. Métodos: Em nosso estudo, investigamos 351 amostras coletadas dos sítios oral, genital e anal de 117 pacientes, por meio da técnica de PCR MY09/11, seguida de genotipagem por RFLP. Todos os pacientes apresentavam lesões genitais benignas. Resultados: A prevalência do HPV foi de 89,7% (105/117) nas lesões genitais, 53,8% (63/117) nas amostras orais e 58,9% (69/117) nas amostras anais. Em relação aos fatores de risco associados à infecção genital, encontramos diferenças estatísticas significativas para prática de sexo oral (p=0,039) e sexo anal (p=0,0000012). Já para as amostras orais, observamos importante correlação entre infecção e uso de contraceptivo oral (p=0,039), tabagismo (p=0,036) e uso de álcool (p=0,0075) enquanto nas amostras anais, alto risco de infecção pelo HPV foi associado a pacientes relatando parceiros sexuais não exclusivos (p=0,013). A presença do DNA viral simultaneamente nos três sítios estudados foi observada em 36,8% dos casos (43/117). Desses, 18% (21/117) apresentaram genótipos concordantes, diferindo da literatura, na qual há grande disparidade de descrições. Conclusão: Há a necessidade de novos estudos a fim de esclarecer a história natural do HPV em sítios extragenitais em diferentes populações, avaliando características anatômicas e fisiológicas com o intuito de esclarecer diferentes taxas de infecção por genótipos do HPV e diferentes processos de doença.

Introduction: Studies concerning human papillomavirus natural history have been focused on cervical infection and disease, but have scarcely described anal infections, especially in clinically health population. Hence, knowledge on HPV natural history is recently being investigated although viral maintenance in hosts is poorly understood. Detection on diverse, extragenital sites may add in the elucidation of infection-dissemination-reinfection cycle reported on the human genital tract. Besides that, it is evident the importance of an adequate screening of the viruses for appropriate diagnosis, mainly in the first steps of the neoplasia in order to provide a better prognosis. Our results pointed out HPV prevalence rates in genital lesions of 87% (80/92) and of 47.8% (44/92) for anal samples. We pointed out that this high prevalence is due to clinically detected lesions, that contributes to the risk of extragenital infections, mainly by host auto-inoculation. Despite the difference in samples size, we did not find statistical relevant differences related with genera (men 85.1% and women 92%; p>0.05), corroborating the idea of no tropism differences by sex. Nevertheless, for anal samples, statistically significant differences were found between men and women (68% of anal HPV infection in women against 40.3% in men (p=0.038). These results reveal that female anal infections are more frequent than in men, suggesting that infection by autoinoculation occurs and can render this site as an HPV reservoir, occasionally becoming clinical lesions. Hormonal profile, sexual behavior and differences among body sites can explain these differences here described. Our study is a preliminary evaluation of HPV infection in human multiple sites.

Introduction: genital herpes is an infectious sexually transmitted disease that affects people from different socioeconomic strata, it is widespread throughout the world and a frequent cause of painful genital lesions in men and women. Objective and Methods: to report a case of a patient with penile ulcers initially treated with various topical and oral products, who finally used Uncaria tomentosa gel topically. The final diagnosis was genital herpes. Results: The symptoms of pain and burning had rapid remission. The clinical course was satisfactory and after a week, the patient, with no lesion, reported having had sexual intercourse without discomfort and returned to professional activities. Conclusion: the topical use of Uncaria tomentosa gel 50 mg/g in penile ulcers was well tolerated, and showed no side effects with relief of local symptoms.

Human papillomavirus (HPV) can cause genital warts and HPV-related cancer.People living with human immunodeficiency virus (HIV)are more symptomatic for HPV infections.Campos dos Goytacazes,a municipality of Rio de Janeiro,introduced the quadrivalent HPV vaccine (4vHPV)for HIV-positive women four years before initiation of a public vaccination program.This study analyzed the prevalence of HPV infection in HIV-positive women and the variables associated with infectionTwo groups were evaluated:group 1,with Pap smear and HPV-negative polymerase chain reaction (PCR);group 2, individuals with at least one positive result for HPV in PCR or pap smear.PCR was performed in endocervical samples using generic primers, and the LCD-Array Kit was used for genotyping.Univariate and multivariate analyzes were performed.Results in 109 women (Group 1 n = 70; group 2 n = 39)showed an overall HPV prevalence of 36%.Results also showed that 88% (n = 23) and 96% (n = 25)of typed viruses (total of typed viruses n = 26)were included in 4vHPV and 9vHPV (nonavalent HPV),respectively.In univariate analysis,age less than 45 years, a high number of sexual partners,and HIV-viral load were risk factors for infection.However, a CD4 indicator was associated with protection.Although HIV infection is generally related to multiple and rare types of HPV,this study showed that a vast majority of the HPV types found are included in 4vHPV.Considering that age less than 45 years is a risk factor, the use of 4vHPV in Brazil should be extended in the public vaccination program to HIV seropositive women up to age 45 years

Introduction: Genital infections by human papillomavirus (HPV) are the most prevalent sexually transmitted viral diseases worldwide. Although the natural history of cervical cancer is better understood, there are still scarce information regarding the etiology of penile cancer, and the natural history of HPV infection in men is not yet fully elucidated. Objective: This study aimed to determine the prevalence of HPV infection in penile samples, from a clinically asymptomatic male population. Methods: A total of 550 samples were collected between January 2011 and July 2014 in different institutions in the State of Rio de Janeiro, including a dermatology clinic and a metallurgical company. The samples were collected from the anatomical regions of the glans and balanopreputial sulcus. HPV identification was made through the generic and type-specific Polymerase Chain Reaction (PCR), and Restriction Fragment Length Polymorphism (RFLP) techniques. Results: An overall prevalence of HPV infection was observed in 21.8% (120 subjects). The most prevalent HPV type was HPV 6 (35%), followed by HPV 16 (20.8%), HPV 11 (19.1%), HPV 31 (6.7%), HPV 33 (6.7%), HPV 45 (8.3%) and HPV 58 (3.3%). Hence, infection was associated with low-risk oncogenic types in 54.1% of the studied individuals, while high-risk oncogenic types were detected in 45.9% of them. The age of the studied subjects ranged from 18 to 65 years with a mean age of 28.4 years. Conclusion: According to our findings, we can infer that the prevalence of HPV infection among asymptomatic male population was considerably lower than the described in the literature, although in agreement with results reported in some recently published studies. We believe that the results may contribute to understand the features of circulation of HPV in male population, in order to evaluate risk-benefits and strategies of disease prevention.

Introdução/objetivos: Infecções pelo ZIKV ocasionalmente podem desencadear um amplo espectro de malformações congênitas, coletivamente denominados de Síndrome da Zika Congênita. Um número restrito de estudos descreve a imunidade na infecção pelo ZIKV durante a gravidez, tanto em modelos experimentais, como em pacientes. Desta forma, buscamos determinar se a resposta imunológica de memória específica ao ZIKV, 2 a 3 anos após a infecção primária, desenvolvida por mães infectadas durante a gravidez e de seus filhos expostos ao vírus por via transplacentária é efetiva e duradoura. Métodos: A resposta efetora de linfócitos T de vinte e uma mães e dezoito crianças foi avaliada por ELISPOT de IFN- γ e citometria de fluxo após estimulação com megapools de ZIKV. Resultados: Como principais achados, observamos uma alta frequência de linfócitos T CD4+ de perfil Th2 efetora/memória e de linfócitos T CD8+ de perfil Th1 naive, seguida de linfócitos T CD8+ de perfil Tc2 efetora/memória nas mães e crianças, indicando que essas células estariam, de alguma forma, auxiliando a resposta imune humoral de mães e crianças com histórico de infecção pelo ZIKV. Observamos ainda, que a capacidade de degranulação e produção de IFN-γ pelos linfócitos T CD4+ foram detectadas nos três grupos de pacientes mesmo após 2-3 anos de infecção, indicando que os linfócitos T CD4+ mantém um perfil de memória de longa duração. Por outro lado, as habilidades de degranulação e produção de IFN-γ pelos linfócitos T CD8+ foram ausentes ou baixos nos três grupos de pacientes após o mesmo período, indicando que os linfócitos T CD8+ mantém um perfil de memória de curta duração quando comparado aos T CD4+. Por fim, demonstramos que os linfócitos T CD4+ TEMRA são os principais produtores de IFN-γ. Conclusão: É importante relembrar que embora não estejamos estudando a infecção aguda na gestação, nossos dados refletem um imprint do que provavelmente ocorreu na infecção aguda. Desta forma, nosso estudo descreve pontos importantes de relevância imunológica, clínica e epidemiológica, particularmente em relação aos linfócitos T CD8+ de memória específicos ao ZIKV que são gerados, mas mantidos por um curto período. Também evidenciarmos que as respostas de linfócitos T específicas ao ZIKV nas mães parecem não ter sido diferenciadas na fase aguda e que, portanto, não estariam relacionadas ao desfecho clínico dos bebês.

Introduction: HPV infection causes cancer at several anatomical sites. However, the infection’s natural history in non-cervical sites is understudied. Objective: To evaluate oral and anogenital HPV infections, correlating HPV prevalence rates and genotypes with site of infection and risk factors. Methods: In the present study, 351 samples from oral, genital, and anal sites of 117 patients were investigated by using PCR MY09/11 detection, followed by genotyping with RFLP. Results: HPV DNA prevalence was 89.7% (105/117) in genital lesions, 53.8% (63/117) in oral samples, and 58.9% (69/117) in anal samples. Regarding the risk factors associated with HPV in genital lesions, statistically significant rates for oral (p=0.039) and anal sex practices (p=0.0000012) were found. For oral samples, a relevant correlation concerning oral contraceptive use (p=0.039), tobacco smoking (p=0.036), and alcohol use (p=0.0075) were observed; whereas in anal samples, higher risk for HPV infection in patients who reported non-exclusive sexual partners (p=0.013) were found. The presence of viral DNA in all the three sites concurrently was observed in 36.8% of the cases (43/117). Among them, 18% (21/117) presented concordant HPV genotypes, diverging from the literature, and thus corroborating that there is still much to learn about HPV natural history, since different biological behaviors are expected within different populations. Differences in anatomy and physiology of the studied sites can determine different prevalence rates of infection by diverse genotypes. Conclusion: Due to the high prevalence of HPV DNA in extragenital sites, further studies are required to define aspects of HPV natural history among different human anatomical sites.

Background: Zika virus (ZIKV) infection causes for mild and self-limiting disease in healthy adults. In newborns, it can occasionally lead to a spectrum of malformations, the congenital Zika syndrome (CZS). Thus, little is known if mothers and babies with a history of ZIKV infection were able to develop long-lasting T-cell immunity. To these issues, we measure the prevalence of ZIKV T-cell immunity in a cohort of mothers infected to the ZIKV during pregnancy in the 2016–2017 Zika outbreak, who gave birth to infants affected by neurological complications or asymptomatic ones.Results: Twenty-one mothers and 18 children were tested for IFN-γ ELISpot and T-cell responses for flow cytometry assays in response to CD4 ZIKV and CD8 ZIKV megapools (CD4 ZIKV MP and CD8 ZIKV MP). IFN-γ ELISpot responses to ZIKV MPs showed an increased CD4 and CD8 T-cell responses in mothers compared to children. The degranulation activity and IFN-γ-producing CD4 T cells were detected in most mothers, and children, while in CD8 T-cells, low responses were detected in these study groups. The total Temra T cell subset is enriched for IFN-γ+ CD4 T cells after stimulation of CD4 ZIKV MP.Conclusion: Donors with a history of ZIKV infection demonstrated long-term CD4 T cell immunity to ZIKV CD4 MP. However, the same was not observed in CD8 T cells with the ZIKV CD8 MP. One possibility is that the cytotoxic and pro-inflammatory activities of CD8 T cells are markedly demonstrated in the early stages of infection, but less detected in the disease resolution phase, when the virus has already been eliminated. The responses of mothers' T cells to ZIKV MPs do not appear to be related to their children's clinical outcome. There was also no marked difference in the T cell responses to ZIKV MP between children affected or not with CZS. These data still need to be investigated, including the evaluation of the response of CD8 T cells to other ZIKV peptides.

There have been reports of neurological abnormalities associated with the Zika virus (ZIKV), such as congenital Zika syndrome (CZS) in children born to mothers infected during pregnancy. We investigated how the immune response to ZIKV during pregnancy is primed and conduct a thorough evaluation of the inflammatory and cytotoxic profiles as well as the expression of CCR5 and CX3CR1. We compared the reactivity of T cells to ZIKV peptides in convalescent mothers infected during pregnancy. The child’s clinical outcome (i.e., born with or without CZS) was taken to be the variable. The cells were stimulated in vitro with ZIKV peptides and evaluated using the ELISPOT and flow cytometry assays. After in vitro stimulation with ZIKV peptides, we observed a tendency toward a higher Interferon gamma (IFN-γ)-producing T cell responses in mothers who had asymptomatic children and a higher CD107a expression in T cells in mothers who had children with CZS. We found a higher frequency of T cells expressing CD107a+ and co-expressing CX3CR1+CCR5+, which is much clearer in the T cells of mothers who had CZS children. We suggest that this differential profile influenced the clinical outcome of babies. These data need to be further investigated, including the evaluation of other ZIKV peptides and markers and functional assays.

BACKGROUND Epigenetic modifications in host cells, like p16 ink4a methylation, have been considered as putative complementary mechanisms for cancer development. Because only a small proportion of infected women develop cervical cancer, other factors might be involved in carcinogenesis, either independently or in association with high-risk human papillomavirus (HR-HPV) infections, including epigenetic factors. OBJECTIVES We hypothesised that p16 ink4a methylation might have a role in cancer development driven by HPV16, mainly in the presence of intact E1/E2 genes. Thus, our objectives were to assess the status of p16 ink4a methylation and the HPV16 E1/E2 integrity in samples in different stages of cervical diseases. METHODS Presence of HPV16 was determined by E6 type-specific polymerase chain reaction (PCR). Methylation status of the p16 ink4a promoter was assessed by methylation-specific PCR in 87 cervical specimens comprising 29 low-grade (LSIL), 41 high-grade (HSIL) lesions, and 17 cervical cancers (CC). Characterisation of E1 and E2 disruption (as an indirect indicator of the presence of episomal viral DNA) was performed by PCR amplifications. FINDINGS We observed a significantly increased trend (nptrend = 0.0320) in the proportion of methylated p16 ink4a in cervical samples during cancer development. Concomitant E1 and E2 disruptions were the most frequent pattern found in all groups: CC (76%), HSIL (54%), and LSIL (73%). No statistically significant differences between p16 ink4a methylation and E1/E2 integrity, in histological groups, was observed. MAIN CONCLUSIONS There was an increase in methylation of the p16 ink4a promoter from pre-neoplastic lesions to cancer. Additionally, a high frequency of E1/E2 disruptions in LSIL/HSIL suggested that viral DNA integration was an early event in cervical disease. Moreover, the methylation status was apparently independent of HPV16 integrity.

ABSTRACTPurpose:The aim of this study was to identify possible protein biomarkers and/or candidates for therapeutic targets in tissues of patients with SCCP, infected by HPV, applying one dimensional electrophoresis (1DE), followed by direct mass spectrometry (MS) analysis.Materials and Methods:Tissues from 10 HPV positive patients with SCCP and from 10 patients with HPV negative non-tumorous penile foreskins were analyzed applying 1D electrophoresis, followed by analysis with direct mass spectrometry (MS).Results:Sixty-three different proteins were identified in the first group and 50 in the second group. Recognition was possible for 28 proteins exclusively detected in Group 1 and 21 proteins presented only in Group 2.Conclusion:Some proteins in the first group are directly involved in the development of other types of cancer, and therefore, suitable for analysis. Complement C3 protein is a strong candidate for evaluating SCCP patients.

SUMMARY High-risk human papillomavirus (hr-HPV) infection is necessary but not sufficient for cervical cancer development. Recently, P16INK4A gene silencing through hypermethylation has been proposed as an important cofactor in cervical carcinogenesis due to its tumor suppressor function. We aimed to investigate P16INK4A methylation status in normal and neoplastic epithelia and evaluate an association with HPV infection and genotype. This cross-sectional study was performed with 141 cervical samples from patients attending Hospital Moncorvo Filho, Rio de Janeiro. HPV detection and genotyping were performed through PCR and P16INK4A methylation by nested-methylation specific PCR (MSP). HPV frequency was 62.4% (88/141). The most common HPV were HPV16 (37%), HPV18 (16.3%) and HPV33/45(15.2%). An upward trend was observed concerning P16INK4A methylation and lesion degree: normal epithelia (10.7%), low grade lesions (22.9%), high grade (57.1%) and carcinoma (93.1%) (p < 0.0001). A multivariate analysis was performed to evaluate an association between methylation, age, tobacco exposure, HPV infection and genotyping. A correlation was found concerning methylation with HPV infection (p < 0.0001), hr-HPV (p = 0.01), HSIL (p < 0.0007) and malignant lesions (p < 0.0001). Since viral infection and epigenetic alterations are related to cervical carcinoma, we suggest that P16INK4A methylation profile maybe thoroughly investigated as a biomarker to identify patients at risk of cancer.

Penile cancer is a potentially mutilating disease. Although its occurrence is relatively rare worldwide, penile cancer rates can be high in developing countries. A few studies have been conducted on the involvement of human papillomavirus (HPV) in penile carcinoma, which have found HPV present in 30-70% of penile malignant lesions, with a higher prevalence of HPV 16 and 18. It has been assumed that cofactors, such as Epstein-Barr virus (EBV) infections, may play a role in the progression of penile neoplasia. The aim of this study was to determine HPV and EBV prevalence in 135 penile malignant lesions from Brazilian men through the use of MY09/11 polymerase chain reaction (PCR), type-specific PCR and restriction fragment length polymorphism analysis. HPV prevalence among the men tested was 60.7%. Of the men who tested positive, 27 presented with HPV 16 (29.7%), five with HPV 18 (5.5%), 21 with HPV 45 (23.1%) and nine with HPV 6 (9.9%). Seven mixed infections were detected (9.2%), while 11 cases remained untyped (13.4%). Regarding EBV positivity, 46.7% of the samples contained EBV DNA with EBV-1 as the most prevalent type (74.6%). More than 23% of the men were co-infected with both HPV and EBV, while 35% presented exclusively with HPV DNA and 20% presented only with EBV DNA. Penile carcinoma aetiology has not been fully elucidated and the role of HPV and EBV infections individually or synergistically is still controversial. Hence, more studies are needed to determine their possible role in carcinogenesis.

INTRODUCTION: The aim of this work was to survey HPV information from a random population of young women from Rio de Janeiro, Brazil. METHODS: This cross-sectional study included cervical samples from 241 female students. To determine human papillomavirus status, polymerase chain reaction amplification was performed. HPV typing was determined by restriction fragment length polymorphism analysis. Demographic data, life style, sexual and gynecological history were obtained through use of a structured questionnaire. RESULTS: The average age of the women was 19.6 years-old (SD=3.4 years). HPV prevalence was 27.4%. Nineteen different HPV genotypes were detected, including 13 high risk types. HPV 16 was the most prevalent type (6.2%), followed by 31 (4.1 %) and 66 (3.7%). Most of the oncogenic types belonged to the A9 species (28/48). The frequency of women infected by at least one oncogenic type was significantly higher than those only infected by low risk types (18.7% versus 7.5%). Cervical changes were detected in 12.5% of the sample and were significantly linked to infection with HPV types of the A9 species. Demographic variables, sexual initiation, or number of sexual partners were not associated with HPV prevalence, variety of HPV genotypes or oncogenic types. CONCLUSIONS: The relative frequency of HPV genotypes other than vaccine types in young females should be taken into account when evaluating vaccination strategies. Due to the high prevalence of HPV infection among the population studied, implementation of sex education in schools, promotion of condom use and an organized screening program to prevent cervical cancer must be encouraged for this age group.INTRODUÇÃO: O objetivo deste trabalho foi investigar a infecção por HPV em uma população randômica de mulheres jovens no estado do Rio de Janeiro, Brasil. MÉTODOS: Este estudo incluiu amostras cervicais de 241 jovens. A detecção de papilomavírus humanos foi realizada pela reação da cadeia da polimerase e a tipificação deste vírus determinada pelo polimorfismo do comprimento do fragmento de restrição. Dados demográficos, estilo de vida, história sexual e ginecológica foram obtidos através de um questionário. A média de idade das mulheres foi 19,6 anos (SD=3,4). RESULTADOS: A prevalência de HPV foi de 27,4% e 19 diferentes tipos foram detectados, incluindo 13 oncogênicos. O HPV 16 foi o tipo mais prevalente (6,2%), seguido do 31 (4,1%) e do 66 (3,7%). A maioria dos tipos oncogênicos pertencia à espécie A9 (28/48). A frequência de mulheres infectadas por ao menos um tipo oncogênico foi significantemente mais alta do que aquelas infectadas por tipos de baixo risco (18,7% versus 7,5%). Alterações cervicais foram detectadas em 12,5% das jovens e foram positivamente relacionadas a tipos de HPV da espécie A9. Variáveis demográficas, iniciação sexual e número de parceiros sexuais não foram associados com a prevalência de HPV, variedade de genótipos ou tipos oncogênicos. CONCLUSÕES: A frequência razoável de outros genótipos além dos vacinais nestas jovens deve ser considerada para avaliação de estratégias de vacinação. Devido à alta prevalência de HPV na população estudada, a implantação de educação sexual nas escolas, incentivo ao uso de preservativo e exames preventivos devem ser encorajados nesta faixa etária.

The objective of the present study was to evaluate the usefulness of molecular methodologies to access human papillomavirus genome in the genital tract. Samples from 136 women aged 17 to 52 years old obtained from the Dr. Sérgio Franco Laboratories between 2000 and 2001, were analyzed by the hybrid capture assay and amplified by PCR with generic primers MY09/MY11 and specific primers for types 16, 18, 31, 33, 35, 58. Viral genome was detected in 71.3% of the samples by hybrid capture and 75% by amplification. When cytopathology was used as a reference method for screening lesions, hybrid capture (p=0) and amplification (p=0.002) presented positive association. The 3 methods showed absolute agreement when cytopathology confirmed papillomavirus infection and high grade intraepithelial lesion. Disagreements occurred for 10 cases: seven inflammatory cases positive by PCR and negative for hybrid capture and 3 low squamous intraepithelial lesions positive for hybrid capture but negative for amplification. In conclusion, hybrid capture was shown to be sensitive and specific enough for use in clinical routines. Moreover, the evaluation of viral load values obtained by this method were shown to be related to the severity of the lesion and merit further studies to analyze the possible association with risk of progression to malignancy.O objetivo do nosso trabalho foi avaliar o emprego de métodos moleculares para comprovar a presença dos papilomavírus humanos no trato genital. Amostras de 136 pacientes com idades entre 17 e 52 anos, coletadas nos Laboratórios Dr. Sérgio Franco entre 2000 e 2001, foram analisadas pelas técnicas de captura híbrida e amplificação pela reação em cadeia da polimerase com primers genéricos MY09/MY11 e específicos para os tipos 16, 18, 31, 33, 35, 58. O genoma viral foi detectado em 71,3% dessas amostras pela captura híbrida e 75% pela reação em cadeia da polimerase. Quando a citopatologia foi usada como método de referência para rastreamento das lesões, a captura (p=0) e a amplificação (p=0,002) demonstraram associação positiva. Os três testes demonstraram concordância absoluta quando a citopatologia diagnosticou processo compatível com papilomavírus ou lesão intraepitelial de alto grau. Dez casos discordantes ocorreram: sete casos de citologia inflamatório positivos na reação em cadeia da polimerase mas negativos na captura híbrida e 3 casos de lesão de baixo grau positivas na captura híbrida e negativas pela reação em cadeia da polimerase. Concluímos que a captura híbrida apresentou sensibilidade e especificidade adequadas para uso clínico. Avaliação da carga viral obtida por esta metodologia relacionou-se com a severidade da lesão e merece estudos adicionais a fim de determinar seu valor prognóstico para o câncer.

INTRODUCTION: Exanthem subitum is a classical rash disease of early childhood caused by human herpesvirus 6B (HHV-6B). However, the rash is frequently misdiagnosed as that of either measles or rubella. METHODS: In this study, a nested multiplex polymerase chain reaction (PCR) was used to diagnose HHV-6B primary infection, differentiate it from infections caused by HHV-6A and compare it to antibody avidity tests. The samples were separated into case group and control group according to the results of the indirect immunofluorescence assay (IFA) technique. RESULTS: From the saliva samples analyzed, HHV-6A DNA was detected in 3.2% of the case group and in 2.6% of the control group. Regarding HHV-6B, PCR detected viral DNA in 4.8% of the case group and in 1.3% of the control group. Among the serum samples studied, a frequency of 1.7% was determined for HHV-6A in the case group and 1.2% in the control group. PCR did not detect HHV-6B DNA in serum samples. The sensitivity and specificity of the PCR technique ranged from 0% to 4.8% and 97.5% to 100%, respectively, compared to IFA. CONCLUSIONS: The PCR technique was not suitable for diagnosing primary infection by HHV-6B in children with exanthematic disease and should not substitute the IFA.

In this study, we assessed the prevalence of human herpesvirus-7 (HHV-7) in 141 serum samples from children less than four years of age with exanthematic disease. All samples were negative for measles, rubella, dengue fever and parvovirus B19 infection. Testing for the presence of human herpesvirus-6 (HHV-6)-specific high avidity IgG antibodies by indirect immunofluorescence assay (IFA) revealed two main groups: one composed of 57 patients with recent primary HHV-6 infection and another group of 68 patients showing signs of past HHV-6 infection. Another 16 samples had indeterminate primary HHV-6 infection, by both IgG IFA and IgM IFA. Serum samples were subjected to a nested polymerase chain reaction to detect the presence of HHV-7 DNA. Among patients with a recent primary HHV-6 infection, HHV-7 DNA was present in 1.7% of individuals; however, 5.8% of individuals tested positive for HHV-7 DNA in the group with past primary HHV-6 infection. Among the 16 samples with indeterminate diagnosis, 25% (4/16) had HHV-7 DNA (p < 0.002). We hypothesise that HHV-7 might be the agent that causes exanthema. However, a relationship between clinical manifestations and the detection of virus DNA does not always exist. Therefore, a careful interpretation is necessary to diagnose a primary infection or a virus-associated disease. In conclusion, we detected HHV-7 DNA in young children from the state of Rio de Janeiro, Brazil.

CONTEXT: Alterations of the p53 tumor suppressor gene are correlated with a critical step in the development of many human cancers.The tumor suppressor gene functions include regulation of the cell cycle and the cellular response to DNA damage, initiation of DNA repair and replication, induction of apoptosis and promotion of cell differentiation. CASE REPORT: Smears from ten cases of cervical lesions were analyzed for status of exons 5-8 of the p53 gene using PCR/SSCP. HPV infection was also screened by the PCR method using two PCR primer sets. Changes in the p53 gene were observed in a case of squamous carcinoma and a case of asymptomatic cervical intraepithelial neoplasia grade III (CIN III). High-risk HPV was detected in both cases showing that HPV infection and p53 mutation are not exclusive events.

Penile carcinoma is an uncommon and potentially mutilating disease with a still unknown etiology. Human papillomavirus (HPV) infection seems to play an important role in the development of a subset of these carcinomas and its presence is thought to be related to the histological type of the lesion. HPV prevalence in penile tumors is reported to be associated to a variety of morphological changes. In recent years, increased insight has been gained into the pathogenesis of penile cancer, the risk factors associated with penile cancer development and the clinical and histological precursor lesions related to this disease. Although penile carcinoma is recognized to be a multi-step process showing a polyclonal profile, a proportion of penile carcinoma is attributable to high risk HPV infection, while in the remaining penile cancers molecular mechanisms independent of HPV are likely to represent the more common underlying events. However, research on the mechanisms behind penile carcinogenesis is warranted.

Os condilomas (verrugas genitais), são lesões hiper-proliferau, as causadas por papilomavírus humanos (HPV) maiona dos condtlomas regridem espontaneamente. mas esões recorrentes são de difícil tratamento. Não há terapia ideal para infecção de papilomavírus na região genital As terapias utilizadas mcluem agentes caústicos, cauterização, cirurgia e inibidores da repcação viral. Entre os antivirais, o interferon (IFN), que também tem propriedades antiproliferativas. e utilizado experimentalmente no tratamento das verrugas. O presente trabalho tem por objetivo o invescgar a ação do IFN em tecido epitelial infectado por HPV. Para verificar se proteínas virais ou celulares tiveram sua sintese afetada por IFN, utilizamos biópsias genitais de pacientes infectados por HPV que não tinham sido submetidos a tratamento prévio. Fragmentos de tecido foram coletados em meio de cultura sem metionina, divididos, e uma das partes tratadas por 106Ul/ml de s-IFN 2b por 24 horas a 37ºC. Após dispersão das células, um pulso de 35S-metionina (5uC/ ui) foi aplicado ao material, durante uma hora a 37ºC. Os extratos foram analisados por eletroforese em gel de poliacrilamida (PAGE) a 12.5% e autorradiografia. A detecção do tipo virai presente nas biópsias foi verificada por hibridização do ADN in situ. Pela análise da incorporação do radiosisótopo, nossos resultados revelaram que os fragmentos de epitélio da pele mantidos in vitro são vitais e que os tratados do PAGE, proteínas de alto peso molecular, provavelmente de origem celular, foram inibidas. Entretanto, ao menos duas proteínas foram induzidas peJo antivirai como visto no PAGE e na autorradiografia O tipo virai 6 foi encontrado em duas biópsias analisadas Outro paciente tinha uma infecção mista pelos upos 6. 11 e 18. O tipo 16 foi detectado em um carcinoma de pênis. Podemos concluir, pelos resultados obtidos, que o IFN promove alterações no perfil de proteínas em biópsias infectadas por HPV. No entanto, são necessários estudos posteriores para determinar a natureza destas proteínas.

CONTEXT: Persistent infection with high risk human papillomavirus (HPV) has been linked to cervical carcinoma. Integration of viral DNA into host cell DNA is essential for this cancer development, promoting disruption of the HPV E2 gene, thus leading to unregulated increases in E6 and E7 proteins and inactivating the products of p53 and Rb tumor suppressor genes. OBJECTIVE: To investigate HPV 16 infection in cervical lesions, physical state of viral DNA and p53 gene alterations in a group of women attending a public health service. DESIGN: Prospective, non-controlled, transversal study. SETTING: Gynecological clinic of the School od Medicine, Universidade Federal Fluminense. SAMPLE: 43 consective patients with cervical lesions referred to our service. MAIN MEASUREMENTS: Cases were classified via cytology/histology as normal, HPV infection, condyloma, low-grade squamous intraepithelial lesion (LSIL), high-grade squamous intraepithelial lesion (HSIL) and carcinoma. HPV infection was studied via polymerase chain reaction (PCR) using two PCR primer sets, to determine DNA integration. p53 gene changes were investigated by single-strand conformation polymorphism (SSCP) analysis. RESULTS: One normal case, 7 HPV infections, 6 condylomas, 7 LSIL, 14 HSIL and 8 cancers were found, with 95% positive for HPV genome when tested using both L1 and E6 primers. HPV 16 was most prevalent (73.1%). HPV 16 DNA was integrated within the host genome in 3 LSIL. One LSIL progressed to HSIL by 13 months after first diagnosis. Among HPV 16-positive HSIL cases, 50% contained integrated viral DNA. HPV 16 E2 gene disruption was seen in 7 cancers (87.5%). Only smal-cell carcinoma showed intact HPV 16 E2 gene. Abnormal p53 bands detected by PCR/SSCP were observed in 4 cases: 2 squamous carcinoma with parametrium (exon 8) and two cervical intraepithelial neoplasia (CIN) III (exons 5 and 7). All cases presented HPV 16 E2 gene loss. CONCLUSIONS: The sample had a high rate of high-risk HPV detected in benign and malignant lesions; high cervical cancer burden; HPV 16 DNA integration in all except one case of cancer; p53 gene changes in CIN III and in invasive cancer cases associated with DNA integration.