Epidemiological surveillance of Human Papillomavirus (HPV) genotypes in oral and genital tracts and the impact of HPV vaccination on male university students from Rio de Janeiro state, Brazil / Vigilância Epidemiológica dos genótipos de papilomavírus humano (HPV) nos tratos oral e genital e o impacto da vacinação em estudantes universitários do sexo masculino do estado do Rio de Janeiro, Brasil

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 por papilomavírus humano (HPV) são de grande preocupação em homens, dada sua possível influência na saúde deles e no risco de transmissão para parceiros. Compreender e abordar essa infecção em homens é fundamental para avaliar a eficácia da vacinação na redução de doenças relacionadas ao HPV. Objetivo: Avaliar o impacto da vacinação contra o HPV, possíveis alterações genotípicas e efeitos adversos, por meio de um estudo prospectivo realizado em estudantes universitários do sexo masculino. Métodos: O estudo envolveu 286 voluntários examinados em Clínicas de Doenças Sexualmente Transmissíveis na Universidade Federal Fluminense, em Niterói, Rio de Janeiro, Brasil. A prevalência do HPV foi avaliada por polymerase chain reaction (PCR) genérico e genotipada por microarranjo de DNA, e foram monitorados os efeitos adversos. Resultados: Os resultados deste estudo revelaram a ausência de efeitos adversos moderados ou graves. Observaram-se mudanças genéticas, incluindo o desaparecimento dos tipos oncogênicos do HPV 16 e 18. Surpreendentemente, mesmo após a conclusão do esquema completo de vacinação, os estudantes ainda abrigavam o HPV 11 na cavidade oral. Além disso, foram identificadas infecções persistentes pelo HPV 6 e 11 em três estudantes que já tinham infecções preexistentes antes da vacinação e na visita de acompanhamento. A análise multivariada revelou associações independentes, especialmente um aumento no risco de infecção pelo HPV na cavidade oral em homens que têm relações sexuais com homens. As taxas de prevalência do HPV permaneceram baixas tanto antes quanto depois do esquema de vacinação (T0: 14,7%, T1: 8,7%). Mesmo após a conclusão do esquema de vacinação, a prevalência permaneceu semelhante em T2 (14,6%), sem diferenças estatisticamente significativas registradas. O HPV 11 emergiu como o tipo mais prevalente ao longo do estudo, seguido pelo HPV 6. Genótipos da vacina foram detectados em uma proporção significativa de amostras em T0 (85,4%), T1 (89,5%) e T2 (100%). Conclusão: No geral, este estudo sugere que a vacinação pode representar uma abordagem promissora para a redução dos riscos à saúde relacionados ao HPV. Esses achados lançam luz sobre os benefícios e desafios potenciais da vacinação contra o HPV, enfatizando a necessidade de monitoramento contínuo e esforços de vacinação

Human Papillomavirus infection in oral and anogenital sites: prevalence and rates of concordance / Infecção pelo Papilomavírus Humano em regiões orais e anogenitais: prevalência e taxa de concordância

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.

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.

Human papillomavirus prevalence, genomic diversity and related risk factors in HIV-positive women from a countryside city in the state of Rio de Janeiro.

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.

Knowledge of human papillomavirus and Pap test among Brazilian university students.

OBJECTIVE: Human papillomavirus (HPV) is the most prevalent sexually transmitted virus in the world and is associated with an increased risk of cervical cancer. The most effective approach to cervical cancer control continues to be screening through the preventive Papanicolaou test (Pap test). This study analyzes the knowledge of university students of health science programs as well as undergraduate courses in other areas of knowledge on important questions regarding HPV. METHOD: Four hundred and seventy-three university students completed a questionnaire assessing their overall knowledge regarding HPV infection, cervical cancer, and the Pap test. A descriptive analysis is presented, and multivariate analysis using logistic regression identified factors associated with HPV/cervical cancer information. RESULTS: Knowledge was higher for simple HPV-related and Pap test questions but was lower for HPV interrelations with genital warts and cervical cancer. Being from the health science fields and having high income were factors associated with greater knowledge. Only the minority of the participants recognized all the situations that increased the risk of virus infection presented in the questionnaire. CONCLUSIONS: These findings highlight the need for educational campaigns regarding HPV infection, its potential as a cervical cancer agent and the forms of prevention available.

Knowledge of human papillomavirus and Pap test among Brazilian university students

SUMMARY OBJECTIVE: Human papillomavirus (HPV) is the most prevalent sexually transmitted virus in the world and is associated with an increased risk of cervical cancer. The most effective approach to cervical cancer control continues to be screening through the preventive Papanicolaou test (Pap test). This study analyzes the knowledge of university students of health science programs as well as undergraduate courses in other areas of knowledge on important questions regarding HPV. METHOD: Four hundred and seventy-three university students completed a questionnaire assessing their overall knowledge regarding HPV infection, cervical cancer, and the Pap test. A descriptive analysis is presented, and multivariate analysis using logistic regression identified factors associated with HPV/cervical cancer information. RESULTS: Knowledge was higher for simple HPV-related and Pap test questions but was lower for HPV interrelations with genital warts and cervical cancer. Being from the health science fields and having high income were factors associated with greater knowledge. Only the minority of the participants recognized all the situations that increased the risk of virus infection presented in the questionnaire. CONCLUSIONS: These findings highlight the need for educational campaigns regarding HPV infection, its potential as a cervical cancer agent and the forms of prevention available.

RESUMO OBJETIVO: O papilomavírus humano (HPV) é o vírus sexualmente transmissível mais prevalente no mundo, estando a infecção por este agente associada a um aumento do risco do câncer de colo uterino. A abordagem mais eficaz para o controle desse tipo de câncer continua sendo a triagem por meio do exame preventivo (Papanicolaou). Este estudo analisa o conhecimento de estudantes universitárias de cursos da área da saúde, bem como cursos de graduação de outras áreas do conhecimento com relação a questões importantes sobre o HPV. MÉTODO: Quatrocentas e setenta e três estudantes universitárias responderam a um questionário que avaliava os conhecimentos sobre a infecção pelo HPV, o câncer de colo do útero e o exame preventivo. Após análise descritiva, foi feita a análise multivariada por regressão logística para identificação dos fatores associados à informação sobre o HPV/câncer de colo do útero. RESULTADOS: O conhecimento das universitárias foi maior para questões simples relacionadas ao HPV e ao exame preventivo, mas foi menor para as correlações do HPV com verrugas genitais e com o câncer de colo do útero. Ser aluna da área da saúde e ter alta renda foram fatores associados ao maior conhecimento. Somente uma minoria das participantes reconheceu todas as situações que aumentavam o risco de infecção pelo HPV apresentadas no questionário. CONCLUSÃO: Os resultados evidenciam a necessidade de realização de campanhas educativas sobre a infecção pelo HPV, do seu potencial como agente de câncer do colo uterino e as formas de prevenção disponíveis.

Investigation of three oncogenic epitheliotropic viruses shows human papillomavirus in association with non-melanoma skin cancer.

Non-melanoma skin cancers (NMSC) share similar risk factors with other virus-related cancers, despite the lack of proved causal association between viral infection and NMSC development. We investigated the presence of Merkel cell polyomavirus (MCPyV), Epstein-Barr virus (EBV), and human papillomavirus (HPV) DNA in 83 NMSC fresh-frozen and 16 non-cancerous skin biopsies and evaluated viral infection according to demographical data, histopathological diagnosis, and ultraviolet exposure. Our results showed that 75% of NMSC biopsies were positive for at least one out of three viruses, whereas only 38% of non-cancerous skin biopsies were positive (p = 0.02). Notably, HPV detection was frequent in NMSC (43%) and nearly absent (one sample, 6.7%) in non-cancerous biopsies (p = 0.007). MCPyV was associated with sites of higher exposure to ultraviolet radiation (p = 0.010), while EBV was associated with a compromised immune system (p = 0.032). Our study showed that HPV was strongly associated with NMSC while EBV and MCPyV with other risk factors. Though further studies are required to elucidate the role of viral infection in NMSC development and management, this study supports the possible role of oncogenic viruses in skin cancers, especially HPV.

The Role of Oncogenic DNA Viruses in Penile Cancer Development.

Penile cancer is a relatively rare neoplasia in developed countries, with significant morbidity and mortality in developing countries. Penile cancer can be subdivided into human papillomavirus (HPV)-positive and HPV-negative cases. Worldwide, the HPV prevalence in penile cancer samples is around 50%, and HPV16 is the most prevalent genotype. Although HPV is an important factor for cancer development, other oncogenic factors may be associated with carcinogenesis. Some of these factors can be infectious, such as the Epstein-Barr virus (EBV), as well as the Merkel cell polyomavirus (MCPyV). The prevalence rates of nearly 50% for both HPV and EBV infections indicate an important role of these viruses in penile tissue malignancy, reinforcing the idea of a multifactorial etiology of the disease. Although the HPV role is better understood, EBV is thought to facilitate persistence, integration, and mutations. Recent studies on the Merkel cell polyomavirus have not shown a relevant prevalence in penile cancer samples, but its presence indicates the opportunistic infectious potential of this virus. Regarding HPV-negative cases, the literature suggests a link with younger age and epigenetic alterations, mainly through the p16INK4a pathway. Recently, several biomarkers that might act as prognostic tools (e.g., Ki-67, squamous cell carcinoma antigen, among others) have been proposed, but the results remain controversial. In addition, other risk factors have also been associated with penile carcinogenesis, such as the presence of phimosis, noncircumcision, chronic inflammation, and number of sexual partners. Further studies are needed to develop tools for early detection and epidemiological surveillance of penile cancer.

Methylation of p16 ink4a promoter is independent of human papillomavirus DNA physical state: a comparison between cervical pre-neoplastic and neoplastic samples

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.

Methylation of p16 ink4a promoter is independent of human papillomavirus DNA physical state: a comparison between cervical pre-neoplastic and neoplastic samples.

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.

Human papillomavirus, Epstein-Barr virus, and methylation status of p16ink4a in penile cancer.

Little is known about penile carcinogenesis. The aim of this study was to evaluate the prevalence of HPV and EBV, and the methylation status of p16ink4a in penile cancer samples, and to contribute to the understanding of the mechanisms responsible for penile cancer development. HPV DNA was detected in 63.6% of 122 cases, with HPV16 being the most prevalent type. EBV DNA was detected in 47.7%, with EBV-1 being the most prevalent type. HPV/EBV co-infections were found in 27.3% of the cases. Hypermethylation in p16ink4a was detected in 64.5% of 110 tested cases. An association between the absence of HPV absence and p16ink4a hypermethylation was also found. Death and/or progressive disease was associated with grade (P = 0.001), T stage (P < 0.0001), and N stage (P < 0.0001). In the multivariable model, grade and N stage were independent risk factors for disease-free survival (P = 0.008 and P < 0.001, respectively). Patients without viral infection had a median age significantly lower than that of the HPV-infected patients. We suggest at least two pathways for penile carcinogenesis, one HPV-independent linked to epigenetic events, probably via p16ink4a inactivation; and another, dependent on HPV infection.

Methylation at 3'LCR of HPV16 can be affected by patient age and disruption of E1 or E2 genes.

CpG methylation at early promoter of HPV16 DNA, in the 3' end of the Long Control Region (3'LCR), has been associated to the presence of episomal forms of viral genome and, consequently, intact E1 and E2 ORFs. The DNA methylation would block the access of E2 viral protein to the E2 binding sites at early-promoter. However, is still unclear if methylation at 3'LCR of HPV16 DNA can also vary depending of other tumor characteristics in addition to viral DNA physical state. In this study, we evaluate whether the methylation level at the five CpG located at 3'LCR of HPV16 is associated to patient age and E1 and/or E2 ORFs integrity. DNA pyrosequencing was used to measure the methylation level in 69 invasive cervical cancer samples obtained from biopsies of patients attended at Brazilian National Institute of Cancer (INCA). PCR amplifications were performed to assess disruption status of E1 and E2 genes of HPV16. The methylation average per sample ranged widely, from <1 to 88.00%. Presence of intact E1/E2 genes and patient age were positively associated with average methylation in both bivariate analyses (p=0.003 and p=0.006, respectively), and multivariate analysis (p=0.002 and p=0.021, respectively), adjusted for tumor type (squamous cell carcinomas or adenocarcinomas) and HPV16 lineage. These findings showed that presence of intact E1/E2 open reading frames was associated with high levels of DNA methylation, and older patients showed higher levels of methylation than younger ones independently of viral genome disruption.

Evaluation of anogenital human papillomavirus infection in asymptomatic men from Rio de Janeiro, Brazil

Infection by human papillomavirus (HPV) is among the most prevalent sexually transmitted diseases worldwide. However, there are still gaps in the knowledge regarding the natural history of HPV infection in men. This study aimed to determine the prevalence of HPV infection in penile swab samples, derived from a clinically asymptomatic male population. For this purpose, 261 samples were collected between January 2011 and July 2013 in different institutions in the city of Rio de Janeiro. These samples were collected from the glans, corona, frenulum and coronal sulcus of the penis. Viral identification was made through generic and type-specific Polymerase Chain Reaction, and Restriction Fragment Length Polymorphism techniques. Prevalence of HPV infection was 16.5% (43 subjects). The most prevalent HPV type was HPV6 (34.9%), followed by HPV16 (23.3%), HPV11 (16.3%), HPV45 (9.3%) and HPV58 (2.3%). Hence, infection was associated with low-risk oncogenic types in 53.7% of the studied individuals, while high-risk oncogenic types were detected in 46.3%. Statistically significant results were found for the group of men who have sex with men, the group who have active anal intercourse, and subjects that lacked circumcision. After adjustments, sexual behavior and lack of circumcision remained as independent risk factors for HPV infection. We believe that these results may contribute to a clearer view about the circulation of HPV in the general male population, as well as to the identification of risk factors associated with the epidemiology of HPV infection in our state.

Proteomics analysis of tissue samples from patients with squamous cell carcinoma of the penis and positive to human papillomavirus.

PURPOSE: 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.

Proteomics analysis of tissue samples from patients with squamous cell carcinoma of the penis and positive to human papillomavirus

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.

Human papillomavirus infection inhealthy men from Rio de Janeiro, Brazil / Infecções por papilomavírus humanos em homens saudáveis do Rio de Janeiro, Brasil

Genital infections by human papillomavirus (HPV) are the most prevalent sexually transmitted viral diseases worldwide. Although the naturalhistory 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 clinicall yasymptomatic male population. Methods: A total of 550 samples were collected between January 2011 and July 2014 in different institutions in the Stateof Rio de Janeiro, including a dermatology clinic and a metallurgical company. The samples were collected from the anatomical regions of the glans andbalanopreputial sulcus. HPV identification was made through the generic and type-specific Polymerase Chain Reaction (PCR), and Restriction FragmentLength 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% ofthem. 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 thatthe prevalence of HPV infection among asymptomatic male population was considerably lower than the described in the literature, although in agreementwith results reported in some recently published studies. We believe that the results may contribute to understand the features of circulation of HPV in malepopulation, in order to evaluate risk-benefits and strategies of disease prevention.

As infecções genitais pelo Papilomavírus humano (HPV) são hoje as mais prevalentes viroses de transmissão sexual em todo o mundo. Embora a história natural do câncer cervical seja melhor compreendida, pouco se sabe sobre a etiologia do câncer de pênis e a história natural do HPV no homem não está completamente elucidada. Objetivo: Este estudo teve como objetivo, determinar a prevalência do HPV em amostras penianas de uma população masculina assintomática. Métodos: Foram coletadas 550 amostras entre janeiro de 2011 e julho de 2014 em diferentes instituições do Estado do Rio de Janeiro, dentre elas:uma clínica de dermatologia e uma indústria metalúrgica. As amostras foram coletadas de sítios anatômicos como sulco balanoprepucional. A identificação do HPV foi feita pela Reação em Cadeia da Polimerase (PCR) genérica e tipo-específica, bem como pelo Polimorfismo do padrão de comprimento de fragmentos de restrição (RFLP). Resultados: A prevalência total da infecção pelo HPV foi de 21,8% (120 indivíduos). O tipo viral de maior prevalência foi o HPV 6 (35%),seguido pelo HPV 16 (20,8%), HPV 11 (19,1%), HPV 31 (6,7%), HPV 33 (6,7%), HPV 45 (8,3%) e HPV 58 (3,3%). A infecção foi majoritariamente associada a tipos de baixo risco oncogênico (54,1%), enquanto os genótipos oncogênicos foram detectados em 45,9% dos pacientes. A idade dos pacientes variou de 18 a 65 anos com média de 28,4. Conclusão: De acordo com nossos achados, podemos sugerir que a prevalência do HPV na população masculina assintomática foi consideravelmente menor do que a descrita em alguns estudos da literatura, mas em acordo com o reportado recentemente por vários autores. Acreditamos que estes resultados podem contribuir para a compreensão dos aspectos epidemiológicos associados à infecção no trato genital masculino, a fim de avaliar estratégias de prevenção de doenças associadas e avaliar o risco-benefício das diferentes abordagens aplicadas em Saúde-Pública.

Genital herpes on the penis and topicuse of uncaria tomentosa: case report / Herpes genital peniano e uso tópico gel de uncaria tomentosa: relato de caso

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.

Herpes genital é uma doença infectocontagiosa de transmissão sexual que acomete pessoas das mais diversas camadas socioeconômicas e está disseminada em todo o mundo, sendo uma causa frequente de lesões genitais dolorosas em homens e em mulheres. Objetivo e Métodos: descrever caso de paciente com úlceras penianas inicialmente medicado com vários produtos tópicos e orais que por último usou, topicamente, gel de Uncaria tomentosa e o diagnóstico final foi herpes genital. Resultados: os sintomas de dor e ardor tiveram rápida remissão. A evolução clínica foi satisfatória e após uma semana o paciente, já sem lesão, informou ter tido relação sexual sem incômodos e retomado as atividades profissionais de forma plena. Conclusão: o uso tópico de gel de Uncaria tomentosa 50 mg/g em úlceras penianas foi bem tolerado, não teve efeitos colaterais com alívio dos sintomas locais.

Human papillomavirus infection in multiple sites / Infecção por papilomavírus humano em múltiplos sítios

Studies concerning human papillomavirus natural history have been focused on cervical infection and disease, but have scarcely describedanal 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 onthe 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. Never heless, for anal samples, statistically significant differences werefound 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 auto inoculation 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 preliminaryevaluation of HPV infection in human multiple sites.

Muitos estudos sobre a história natural do papilomavírus humano (HPV) têm focado nas infecções cervicais, mas pouco tem se falado sobre infecções anais em pacientes assintomáticos. Assim, a história natural do HPV ainda está em construção e os mecanismos de manutenção dele no organismo hospedeiro são pouco compreendidos. Entendemos que a detecção do vírus em sítios extragenitais poderá ajudar na compreensão da cadeia de infecção disseminação-reinfecção pelo HPV no trato genital humano. Além disso, é evidente a importância de um rastreio adequado do vírus diante de diagnósticos clínicos, principalmente no período inicial da lesão, provendo, assim, um melhor prognóstico. Nossos resultados apontaram uma prevalência da infecção por HPV em lesões genitais de 87% (80/92) e nas amostras anais tal prevalência foi de 47,8% (44/92). Ressaltamos que nossa prevalência foi alta, mas estudamos indivíduos com lesões genitais clinicamente detectadas, aumentando, assim, o risco de infecção em outros sítios e confirmando a ocorrência da presença do HPV no hospedeiro, provavelmente decorrente de autoinoculação. Apesar da diferença entre o número de indivíduos dos sexos feminino e masculino em nosso estudo, não encontramos diferenças estatísticas relacionadas à presença do HPV em lesões genitais (H: 85,1% e M: 92%; p>0.05),corroborando que não há diferenças entre o tropismo por gênero referente às infecções genitais. Entretanto, no caso das infecções anais, a porcentagem foi estatisticamente diferente: 68% de HPV anal em mulheres e 40,3% em homens (p=0,038). Estes resultados apontam que a aquisição da infecção anal em mulheres, apesar de assintomática, é mais frequente do que em homens, sugerindo que esta mucosa possa ser infectada por autoinoculação e funcionar como um reservatório feminino, o que eventualmente poderá resultar em lesões. Tais discrepâncias da prevalência entre homens e mulheres poderiam se explicadas pelo comportamento sexual, por efeitos hormonais e por diferenças entre sítios. Nosso estudo se trata de um levantamento preliminar da infecção por HPV em múltiplos sítios.

An upward trend in DNA p16ink4a methylation pattern and high risk HPV infection according to the severity of the cervical lesion.

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.

An upward trendiin dna p16ink4a methylation patern and high risk hpv infectionn according to the severity of the cervical lesion / Padrão de metilação do gene P16INK4A e de infecções por HPV de alto risco acompanham a evolução de lesões cervicais

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. .

RESUMO É reconhecido que infecções por papilomavírus humanos de alto risco (HPV) são causa necessária, mas não suficiente para o desenvolvimento do câncer cervical. Recentemente, estudos de silenciamento gênico apontaram que a hipermetilação do gene p16INK4A é importante co-fator para a carcinogênese cervical, eliminando a função supressora de tumor da proteína p16 em lesões malignas. Entretanto poucos estudos avaliaram a relação da metilação com a progressão da doença. Nosso objetivo foi investigar o padrão de metilação do gene P16INK4A em diferentes graus de lesão cervical e sua associação com a infecção por diferentes tipos de HPV. Nosso estudo de corte transversal avaliou 141 amostras cervicais de pacientes atendidas no Hospital Moncorvo Filho, Rio de Janeiro. A detecção e tipagem do HPV foi realizada pela técnica de reação em cadeia da polimerase (PCR), e a metilação do gene P16INK4A pela PCR-metilação específica em formato nested (MSP). A frequência de HPV foi de 62,4% (88/141). O tipo mais prevalente foi o HPV16 (37%), seguido pelo HPV18 (16,3%) e HPV33/45 (15,2%). Curva ascendente foi observada quanto ao padrão de metilação do gene P16INK4A e o grau da lesão: a metilação foi identificada em somente 10,7% das amostras de epitélio normal, em 22,9% das lesões de baixo grau, em 57,1% das lesões de alto grau e em 93,1% dos carcinomas (p < 0,0001). Foram feitas análises univariada e multivariada a fim de correlacionar metilação, idade, exposição ao tabaco, infecção e genótipo de HPV. Foi encontrada correlação da metilação com a infecção pelo HPV (p < 0,0001), genótipos de alto risco (p = 0,01), ...

Detection of Merkel cell polyomavirus in oral samples of renal transplant recipients without Merkel cell carcinoma.

Merkel cell carcinoma (MCC) is a rare but aggressive neuroendocrine cancer, with approximately 80% of cases associated with Merkel cell polyomavirus (MCPyV). The lack of information concerning its occurrence in non-MCC immunosuppressed populations led to the investigation of MCPyV DNA in saliva and oral biopsies from 60 kidney allograft recipients and 75 non-transplanted individuals (control group). In contrast to herpesviruses, which was also investigated (CMV, HHV-6A, and B, HHV-7) MCPyV was detected predominantly in patients with oral lesions (gingivitis and/or periodontitis) of both transplanted and non-transplanted groups (P=0.016) and in the saliva of the transplanted group (P=0.009). MCPyV co-detection with CMV (P=0.048), and HHV-6 (P=0.020) in the saliva of transplanted patients requires further investigation on a possible role of co-infection.