Prevalencia de los genotipos de HPV en lesiones pre invasoras de alto grado de malignidad y cáncer de cuello uterino en la población del Hospital de Clínicas: Montevideo-Uruguay / Prevalence of HPV genotypes in cervical high risk intraepithelial lesions and cervical cancer in the population of Hospital de Clínicas: Montevideo-Uruguay / Prevalência de genótipos de HPV em lesões pré-invasivas com alto grau de malignidade e câncer de colo do útero na população do Hospital de Clínicas: Montevidéu, Uruguai

El Cáncer de Cuello Uterino (CCU) es un problema de Salud pública a nivel mundial. Su indiscutible asociación con el Virus del papiloma humano (HPV) hace necesario su estudio. El objetivo de este trabajo, es conocer la prevalencia de los diferentes genotipos de HPV, en lesiones pre invasoras de alto grado de malignidad (HSIL) y/o cáncer de cuello uterino. Material y Métodos: Todas las Mujeres que fueron derivadas a pol de TGI del H Clínicas entre enero del 2011 y diciembre de 2012, por un PAP sospechoso de lesión y en las que se confirmó luego un HSIL o cáncer cervical fueron tipificadas. Se recabaron datos de edad, tipo de lesión y genotipificación. La extracción de ADN viral se realizó a partir de muestras cervico vaginales conservadas en medio de transporte comercial (Sacace) mediante el kit QIAamp DNA Mini Kit (QIAGEN). Se buscaron 14 genotipos de alto riesgo. Resultados: Se tipificaron 75 pacientes, 19 con CCU y 56 con HSIL. El HPV 16 fue el más prevalente en un 61,5 % para las infecciones únicas y en un casi 100 % para las múltiples y un 60% para el total de las lesiones, seguido en prevalencia por los HPV 31,33 y 45. El HPV 18 fue muy poco prevalente. Conclusiones: En esta muestra, la prevalencia del HPV 16 está acorde con las publicaciones nacionales siendo el más frecuente. El HPV 18 tiene muy baja prevalencia siendo 2 casos en 75, siempre en infecciones múltiples.

Cervical Cancer (CC) is a public health problem worldwide. Its indisputable association with the Human Papilloma Virus (HPV) makes its study necessary. The objective of this work is to know the prevalence of the different HPV genotypes, in pre-invasive high-grade malignant lesions (HSIL) and / or cervical cancer. Material and Methods: All women who were referred to Low genital tract service in the Hospital de Clinicas between January 2011 and December 2012, for a PAP suspected of injury and in which HSIL or cervical cancer was later confirmed were typified. Data on age, type of lesion and genotyping were collected. The viral DNA extraction was carried out from cervico-vaginal samples preserved in commercial transport medium (Sacace) using the QIAamp DNA Mini Kit (QIAGEN). 14 high-risk genotypes were searched. Results: 75 patients were typified, 19 with CCU and 56 with HSIL. HPV 16 was the most prevalent in 61.5% for single infections and almost 100% for multiple infections and 60% for all lesions, followed in prevalence by HPV 31,33 and 45. The HPV 18 was very rare. Conclusions: In this sample, the prevalence of HPV 16 is in accordance with national publications, being the most frequent. HPV 18 has a very low prevalence, being 2 cases in 75, always in multiple infections.

O câncer cervical é um problema de saúde pública em todo o mundo. E um câncer com uma ligação comprovada com o vírus do papiloma humano. O objetivo é conhecer a prevalência dos diferentes genótipos do HPV em mulheres que apresentam neoplasias pré-invasivas de alto grau e câncer de colo do útero, que foram tratadas e diagnosticadas no Hospital de Clínicas entre janeiro de 2011 e dezembro de 2012. Material e Métodos: o estudo foi realizado em 75 pacientes do Hospital de Clinicas, com diagnóstico histológico de Câncer Cervical e lesões intraepiteliais de alto grau para as quais foi realizado o tipageme do HPV alto risco. Resultados: 75 pacientes foram tipificados, 19 com CCU e 56 com HSIL. O HPV 16 foi o mais prevalente em 61,5% para infecções únicas e quase 100% para infecções múltiplas e 60% para todas as lesões, seguido em prevalência pelo HPV 31,33 e 45. O HPV 18 era muito raro. Conclusões: Nesta amostra, a prevalência do HPV 16 está de acordo com as publicações nacionais, sendo a mais frequente. O HPV 18 tem prevalência muito baixa, sendo 2 casos em 75, sempre em infecções múltiplas.

High-risk human papillomavirus and cervical lesions among women living with HIV/AIDS in Brazilian Amazon, Brazil

ABSTRACT OBJECTIVE: The goal of this study was to determine the prevalence of human papillomavirus infection infection and cervical lesions and its associated factors among HIV infected women attending an AIDS clinic in Amazonas state, Brazil. METHODS: Cross-sectional study. Women attending an AIDS clinic in the city of Manaus between March and December 2011 for gynecological examination were invited to participate. Enrolled patients answered a standardized interview including demographical, behavioral, and clinical data. Additionally, patients underwent a gynecological evaluation with collection of cervical samples for cytological analysis and high-risk human papillomavirus infection hybrid capture. A blood sample was also obtained to determine CD4 and viral load levels. RESULTS: A total of 310 (82.9%) women participated in the study. High-risk human papillomavirus infection was detected in 191 (61.6%) cases; 24 (13.5%) had low-grade squamous intraepithelial lesion (SIL) and 4 (2.2%) high-grade SIL. No invasive cervical cancer was diagnosed. Median age was 32 (interquartile range (IQR): 27-38) years and median of education was 8.5 (IQR 4-11) years of schooling and 56.1% had a monthly income up to US$180. In multivariate analysis, being less than 30 years old [OR = 1.7 (95% CI: 1.2-2.4,p = 0.005)], high-grade SIL [OR = 6.5 (95% CI: 1.6-23.0, p = 0.009)], and CD4 counts <200 cells/mm3 [OR = 1.6 (95% CI: 1.2-2.0, p < 0.001)] were associated with high risk human papillomavirus infection infection. CONCLUSIONS: In the present study high-risk human papillomavirus infection was frequent and it was associated to high-SIL. These results show the importance of gynecologic examinations in routine care and follow-up required by those who present with cervical lesions.

Human papillomavirus risk factors for infection and genotype distribution in aboriginal women from Northern Argentina / Virus papiloma humano (hpv) en mujeres de la etnia Pilagá del nordeste argentino: Factores de riesgo de infección y distribución de tipos virales

The mortality rate for cervical cancer (CC) in Northern Argentina is three times higher than the average for the country (7.8 deaths/100 000 women). We determined the prevalence and genotype distribution of human papillomavirus (HPV) in 227 sexually active women of the native Pilagá community in Formosa, Argentina. We also conducted an HPV-16 variant analysis and studied several community factors that might play a role in viral entry and infection. Endo- and exocervical samples were tested for HPV DNA with MY09/11-PCR or with GP5+/6+-PCR. HPV was detected in 46.7% of the samples and 21 different types were found; the most frequent being HPV-16 (19.4%), -6 and -18 (5.3%), -58 (3.5%) and -31 and -33 (3.1%). In relation to HPV-16 variants, 68.2% were European and 31.8% Asian-American. Among the cofactors analyzed only disposal of human excreta to the open air (P=0.01) was significantly associated with HPV infection. Our prevalence estimates clearly show that Pilagá women are highly exposed to or infected with high risk HPV types and therefore are at a high risk of developing precancerous lesions and eventually CC at the population level.

La tasa de mortalidad por cáncer cervical (CC) en la región norte de la Argentina es tres veces más alta que la media del país (7.8 muertes/100 000 mujeres). En el presente trabajo se determinó la prevalencia de infección por virus papiloma humano (VPH) y la distribución y frecuencia de los genotipos en 227 mujeres sexualmente activas de la etnia aborigen Pilagá (Formosa, Argentina). También se realizó un análisis de las variantes intratípicas de VPH-16 presentes en la comunidad y se analizaron diversos factores socioculturales que podrían tener algún rol destacado en la transmisión de la infección viral. Se estudiaron muestras de células endo-exocervicales mediante PCR basadas en los cebadores MY09/11 y GP5+/6+ con posterior restricción enzimática y/o hibridación dot-blot. La infección por VPH fue detectada en el 46.7% de las mujeres analizadas. Fueron identificados 21 genotipos, de los cuales los más frecuentes fueron HPV-16 (19.4%), -6 y -18 (5.3%), -58 (3.5%) y -31 y -33 (3.1%). Respecto al HPV-16, se encontraron 68.2% de variantes europeas y 31.8% de asiático-americanas. Entre los cofactores analizados, solo la disposición de excretas al aire libre estuvo significativamente asociada con la infección por VPH (P = 0.01). Los datos obtenidos reflejan que la comunidad Pilagá está altamente expuesta a las infecciones por genotipos de alto riesgo de VPH, lo cual puede estar asociado a una alta incidencia de lesiones cervicales preneoplásicas y neoplásicas.

HPV infection and intraepithelial lesions from the anal region: how to diagnose?

In the last years, the prevalence of HPV infection in the anal region has increased, especially in some groups like homosexual and HIV-positive people. Since this infection can be associated with the development of squamous anal cancer due to its progression from HPV infection to anal intraepithelial neoplasia (AIN) and finally to cancer, the screening and evaluation of these conditions are important. Anal cytology and high resolution anoscopy are good methods that are available and can be used. Although useful, these methods should be performed correctly and not indiscriminately in all patients. Patients for whom anal cytology screening is recommended are: HIV-infected patients, homosexuals, women who present with high-grade vulvar squamous intraepithelial neoplasia, vulvar cancer or cervical cancer. An abnormal anal cytology should be further evaluated with high resolution anoscopy.

Melanosis of the vagina and human papillomavirus infection, an uncommon pathology: Case Report / Melanosis de la vagina e infección por virus del papiloma humano, una patología poco común: Reporte de un Caso

Benign melanotic lesions of the vagina are uncommon and only a few cases have been reported in the literature. A 34-year-old woman was referred because of a Vaginal Intraepithelial Neoplasia 1 biopsy result. On the gynecological examination, two different hyperpigmented areas were noted in the vagina. The colposcopic visualization of the cervix and vagina found an aceto-white lesion at the right lateral wall of the upper third of the vagina. Biopsies from three areas were taken. Histological study reported a melanosis of the vagina and HPV infection. An immunohistochemical panel of epithelial markers was performed in vaginal samples, such as Cytokeratin AE1/AE3 and epithelial membrane antigen, mesenchymal marker: vimentin; melanocytic makers: protein S-100 and HMB45 (Human Melanoma Black); proliferating cell marker: proliferating cell nuclear antigen (PCNA), and P-53 oncoprotein. High Risk (16, 18, 31, 45) and Low Risk (6, 11) HPV types were studied by In Situ Hybridization using the same vaginal samples. CK, EMA and Vimentin were 2+. Melanocytic markers, HMB45 and S100, and PCNA were 1+ in basal cell layer. P-53 was negative. The melanotic tissue and acetowhite lesion were positives to HPV Types 6,11. In conclusion, melanosis of the vagina is a uncommon benign pathology. Usually, melanosis is present in women over 40 years old. We present a case of melanosis of the vagina in a young woman infected with low-risk HPV types and review the literature.

Las lesiones melanóticas de la vagina son infrecuentes. y Solo pocos casos han sido reportados. Se presenta el caso de una mujer de 34 años quien es referida con diagnóstico de una Neoplasia Intraepitelial Vaginal 1. Al examen ginecológico, se encontraron dos áreas hiperpigmentadas en la vagina. La exploración colposcópica del cuello uterino y vagina reveló la presencia de una lesión aceto-blanca en la pared lateral derecha del tercio superior de la vagina. Muestras de biopsias fueron tomadas en dichas áreas. El estudio histológico reportó una melanosis de la vagina y una infección por el virus del Papiloma Humano (VPH). Se realizó un panel de estudio inmunohistoquímico de marcadores epiteliales en las muestras vaginales: tales como citoqueratina AE1/AE3 y antígeno epitelial de membrana; marcador mesenquimal: vimentin; marcadores melanóticos: proteina S-100 y HMB45 (Human Melanoma Black); marcadores de proliferación celular: antígeno de proliferación nuclear (PCNA), y la oncoproteína P-53. Se realizó Hibridización In Situ para establecer los tipos de alto (16, 18, 31, 45) y bajo (6, 11) riesgo de VPH en las muestras vaginales. Los marcadores CK, EMA y Vimentin fueron 2+. Los marcadores melanótico, el HMB45 y el S100, y el PCNA fueron 1+ en la capa basal. P-53 fue negativo. El tejido melanótico y la lesión acetoblanca fueron positivos al VPH 6,11. En conclusión, la melanosis vaginal es una patología poco frecuente. Usualmente, se ha reportado en mujeres mayores de 40 años. Presentamos un caso de una melanosis de la vagina infectada con un tipo de VPH de bajo riesgo en una mujer joven y una revisión de la literatura.

Short-term risk of cervical intraepithelial neoplasia grades 2 and 3 for women with normal cytology and human papillomavirus infection / Riesgo a corto plazo de lesiones intraepiteliales cervicales grados 2 y 3 en mujeres con citología vaginal normal e infección por el virus del papiloma humano

OBJECTIVE. To assess the risk of cervical intraepithelial neoplasia grades 2, 3 or higher (CIN 2/3+) for women with normal cytology and concurrent high-risk human papillomavirus infection (HR-HPV). MATERIAL AND METHODS. We examined 2 200 women every 6 months for an average of 9 years. Cervical smears and samples for HPV DNA were obtained at each visit. Absolute risk of subsequent CIN2/CIN3+ was estimated using the Kaplan-Meier method. RESULTS. The absolute risk of CIN2/CIN3+ among HR-HPV-positive women with normal Pap smear results was 1.06 percent (95 percentCI, 0.57-2.20), 5 times higher the risk among all women with normal Pap smears (0.20 percent; 95 percentCI, 0.12-0.32) but 7 times lower than that for women with HR-HPV infection and LSIL (7.24 percent; 95 percentCI, 3.78-15.2). CONCLUSION. Short-term absolute risk of CIN2/3+ after a normal Pap smear with concurrent HR-HPV infection is low (~1 percent), suggesting that the HR-HPV test has limited utility in short-term clinical decision-making for women with normal cytology.

OBJETIVO. Evaluar el riesgo a corto plazo de neoplasia intraepitelial cervical de alto grado (CIN2/CIN3+) en mujeres con citologí-a cervicouterina normal e infección por virus del papiloma humano de alto riesgo (HR-HPV). MATERIAL Y MÉTODOS. Cohorte prospectiva de 2200 mujeres evaluadas cada seis meses durante 9 años en promedio. En cada visita se tomó muestra cervical para extendido y detección de HPV DNA. El riesgo absoluto de CIN2/CIN3+ a la siguiente visita fue calculado utilizando el método de Kaplan-Meier. RESULTADOS. En mujeres con citologí-a normal e infección concomitante por HR-HPV el riesgo absoluto de presentar CIN2/CIN3+ fue de 1.06 por ciento (95 por cientoCI, 0.57-2.20). Este riesgo fue cinco veces mayor al observado en todas las mujeres con citologí-a normal (0.20 por ciento; 95 por cientoCI, 0.12-0.32) pero siete veces menor que el observado en mujeres con lesiones intraepiteliales escamosas de bajo grado con infección concomitante (7.24 por ciento; 95 por cientoCI, 3.78-15.2). CONCLUSIÓN. El riesgo absoluto de CIN2/3+ a corto plazo luego de una citologí-a normal e infección por HR-HPV es baja (~1 por ciento), sugiriendo que, a corto plazo, la prueba de HR-HPV tiene utilidad clí-nica muy limitada en mujeres con citologí-a normal.

Nuevos paradigmas y desafíos en la prevención y control del cáncer de cuello uterino en América Latina / New paradigms and challenges in cervical cancer prevention and control in Latin America

El cáncer de cuello uterino sigue siendo un problema de salud pública en Latinoamérica. El uso de la citología para la detección de lesiones pre-cancerosas no ha tenido mayor impacto en las tasas de incidencia y mortalidad, que aún se mantienen altas en la región. La disponibilidad de nuevas técnicas de tamizaje para la detección de lesiones pre-cancerosas y de vacunas altamente eficaces que previenen casi todas las lesiones relacionadas con VPH-16 y VPH-18 en mujeres no expuestas previamente al virus representan una gran oportunidad para la prevención del cáncer de cuello uterino en la región. En este manuscrito resumimos la evidencia científica y la experiencia de la región en i) el uso de pruebas de VPH y de la inspección visual después del ácido acético (IVAA) en tamizaje primario, y ii) la implementación de programas de vacunación en adolescentes. Finalmente enumeramos una serie de recomendaciones adecuadas para distintos escenarios. La factibilidad de implementar un programa nacional de prevención de cáncer de cuello uterino exitoso y sostenible en países latinoamericanos dependerá de las prioridades de salud, la infraestructura y personal de salud disponible, determinadas luego de un riguroso análisis situacional local.

Cervical cancer continues to be a significant health problem in Latin America. The use of conventional cytology to detect precancerous cervical lesions has had almost no major impact on reducing cervical cancer incidence and mortality rates, which are still high in the region. The availability of new screening tools to detect precancerous lesions provide great opportunities for cervical cancer prevention in the region, as do highly efficacious HPV vaccines able to prevent nearly all lesions associated with HPV-16 and -18 when applied before viral exposure. This paper summarizes the scientific evidence and regional experiences related to: i) the use of HPV testing and visual inspection after the application of acetic acid (VIA) in primary screening and ii) the implementation of adolescent HPV vaccination programs. Finally, we outline a number of recommendations for different resource settings. The feasibility of implementing successful and sustainable national cervical cancer prevention programs in Latin American countries in the region will depend on health priorities and the availability of infrastructure and health personnel-as determined by rigorous local situational analysis.

Detección y tipificación de virus papiloma humano en lesiones preneoplásicas de cuello uterino / Human papillomavirus genotyping of cervical uterine preneoplastic lesions in a high risk area

The relationship between human papillomavirus (HPV) and uterine cervical cancer (UCC) is widely known and accepted. Aim: To determine the frequency of genotypes of HPV in cervical preneoplastic lesions in a high risk area of UCC. Material and Methods: Using a combination of PCR and Reverse Line Blot technique, 235 formalin fixed paraffin embedded samples, with diagnosis of low-grade squamous intraepithelial lesion (LSIL) or high-grade squamous intraepithelial lesion (HSIL) were genotyped. Results: HPV was detected in 61.2 percent of LSIL and 78.1 percent of HSIL. The main genotypes found were HPV 16, 18, 31, 45, 56 y 58. HPV 16 was the most common in both LSIL (18.1 percent) and HSIL (36.9 percent). HPV 16 or 18 were present in 25.1 percent and 47.1 percent of the LSIL and HSIL respectively. In both LSIL and HSIL, the predominant viral genotypes were those types classified as with a high oncogenic risk. Conclusions: HPV genotypes 16, 18, 31, 45, 56 y 58 were the most common in our series. HPV 16 and 18, viral types with high oncogenic risk and included in commercial vaccines, were found in 25.1 percent and 47.1 percent of LSIL and HSIL, respectively.

High prevalence and low E6 genetic variability of human papillomavirus 58 in women with cervical cancer and precursor lesions in Southeast Mexico

Infection with some genotypes of human papillomavirus (HPV) is the most important risk factor associated with cervical cancer (CC). Throughout the world, HPV type 58 prevalence varies from one region to another; it is higher in women from certain countries in Asia and Latin America, such as China and Mexico. Although intratypic variants have been reported on a few occasions, our knowledge about HPV 58 genetic variation remains limited. Therefore, this work aims to (i) determine the prevalence of HPV type 58 amongst Mexican women with invasive CC or precursor lesions and (ii) identify HPV 58 sequence variants. One hundred and forty five colposcopy clinic patients were studied. Genotyping of HPV 16, 18 and 58 was determined by specific nested PCR and HPV 58 variants were detected by direct sequencing. The general prevalence of HPV was 51.7 percent (75/145). HPV 16 was found in 30.6 percent (23/75) and HPV 58 in 24 percent (18/75) of the patients. HPV 18 was not identified in patients with cervical intraepithelial neoplasia (CIN) grade I; it was only found in those with CIN II, with a prevalence of 6.8 percent (3/44). In patients with CC, the prevalence of HPV 16 and 58 was 78.9 percent. Regarding HPV 58 variants, 94.4 percent of the HPV 58 sequences were identical to the prototype strain, whereas one sample showed changes at a single nucleotide. This study demonstrates a high prevalence of HPV 58 and a low genetic variability of E6 sequences amongst Mexican colposcopy patients.

Prevalência do herpes-vírus humano tipo 1 em neoplasias cutâneas epiteliais malignas / Prevalence of human herpes virus type 1 in epithelial skin cancer

FUNDAMENTOS - O DNA viral pode atuar como oncogene, favorecendo o desenvolvimento de neoplasias, como as linfoides e da pele. Entre esses vírus, encontram-se alguns herpes-vírus humanos. OBJETIVO - Identificar a presença de DNA do herpes-vírus humano tipo 1 em neoplasias epiteliais pré-malignas,malignas e pele normal de indivíduos controle, avaliando seu papel na carcinogênese. MÉTODOS - Identificação, por reação em cadeia da polimerase, do DNA viral do tumor e pele sã de 41 pacientes e comparação com grupo controle, sem neoplasia. Análise estatística: Testes de Fisher e de McNemar. RESULTADOS - O vírus foi identificado em 20 indivíduos sem e em 21 com neoplasia. Destes últimos, 11 o expessaram apenas nas células tumorais. A diferença, entretanto, não foi estatisticamente significante. CONCLUSÕES - Parece não haver relação direta entre o encontro do DNA viral na pele sã e na pele tumoral. Sua presença pode facilitar o desenvolvimento da neoplasia ou apenas coincidir de se localizar onde esta já ocorreu.

BACKGROUND - Viral DNA may act as an oncogene, especially in skin and lymphoid organs. This group includes some human herpes virus. OBJECTIVE - To identify human herpes virus type 1 DNA in pre-malignant and malignant skin samples of epithelial tumors comparing to normal skin to determine its role in carcinogenesis. METHODS - Forty-one patients with epithelial tumors were submitted to biopsies from tumor and normal skin. The control group comprised 41 biopsies from patients with other dermatoses than cancer. After DNA extraction, polymerase chain reaction was performed to identify 199-bp band. The results were statistically evaluated by Fisher and McNemar tests. RESULTS - The virus was identified in 20 subjects without cancer and in 21 with skin cancer. From these, 11 expressed it only in tumor cells. This difference was not significant. CONCLUSION - There seem to be no direct relation between viral findings in normal skin and skin cancer cells. It may act as a promoter or just coexist at the same site where a neoplastic transformation has already occurred.

Quanto a escova deve ser introduzida no canal anal para avaliação citológica mais eficaz? / How deep must the brush be introduced in the anal canal for a more effective cytological evaluation?

OBJETIVO: A sensibilidade da citologia anal varia amplamente na literatura, entre 45 por cento e 98 por cento, o que pode ocorrer devido à falta de padronização quanto à distância que a escova deve ser introduzida no canal anal. Desta forma, nosso objetivo foi investigar se o local de coleta influencia no resultado desse exame. MÉTODOS: Colhemos amostras com escova introduzida 4 cm no canal anal de 114 doentes (Grupo A) e 2 cm em outros 94 pacientes (Grupo B), antes do exame proctológico. Realizamos cinco rotações com a escova antes de retirá-la e a esfregar sobre lâmina de vidro, posteriormente submetida ao exame citopatológico padrão. Todos os doentes são HIV-positivo. Submetemos os resultados à avaliação estatística. RESULTADOS: No Grupo A, 39 doentes possuíam condilomas no canal anal e a citologia foi positiva em 29 deles (74,3 por cento). Também observamos alterações citológicas em 30 de 75 doentes (40 por cento) sem lesões clínicas no canal anal. No Grupo B, havia 54 doentes com condilomas no canal anal e em 13 (24,1 por cento) houve confirmação citológica. Em 40 outros, sem lesões clínicas pelo HPV, notamos que em nove (22,5 por cento) havia anormalidades citológicas. Os testes estatísticos revelaram que os exames realizados nos doentes do Grupo A foram mais eficientes. CONCLUSÃO: Os espécimes coletados com escovas inseridas mais profundamente no canal anal melhoraram a eficácia do exame.

OBJECTIVE: In literature, sensitivity of Pap smears ranges widely from 45 to 98 percent. Possibly this is because there is no standard for how far the brush should be introduced into the anal canal. The aim was to evaluate whether the sampling site has an influence on the results of anal Pap smears. Design and setting. This is a non-randomized, non-blinded, retrospective review carried out in the Proctology and Pathology Sectors, Emilio Ribas Infectious Diseases Institute. METHOD: We obtained specimens with brushes introduced 4 cm into the anal canal in 114 patients (Group A) and 2 cm in 94 patients (Group B), before anorectal examination. These brushes were rotated five or six times before being withdrawn and rubbed on a slide that underwent Pap testing using standard cytopathology laboratory equipment. All patients were HIV-infected. Statistical tests were used. RESULTS: In Group A, 39 patients had anal canal condylomas and the cytology was positive in 29 of them (74.3 percent). We also observed cytological alterations in 30 of 75 patients (40 percent) without clinical lesions in the anal canal. In Group B, there were 54 patients with condylomas and 13 of them (24.1 percent) were confirmed by cytology. In 40 patients with no clinical lesions, we observed that nine (22.5 percent) had cytological abnormalities Statistical analysis revealed that examination in Group A was more efficient. CONCLUSION. Specimens collected by inserting the brush deeper into the anal canal improved the efficiency of anal Pap smears.

p16INK4 expression in precursor lesions of squamous cell cervical cancer related to the presence of HPV-DNA

The purpose of the present study was to identify the expression of p16INK4 in cervical cancer precursor lesions by immunohistochemistry and to correlate it with lesion grade and presence of human papillomavirus (HPV) infection. Cervical specimens from 144 women seen consecutively at the gynecology outpatient clinic of our institution from December 2003 to May 2005 were analyzed by cytopathology, histopathology, polymerase chain reaction for HPV-DNA, and p16INK4 immunostaining. Histologically normal biopsies, HPV-DNA negative by polymerase chain reaction, were used as control. HPV-DNA prevalence, including the control group, was 68.1 percent and the prevalence of p16INK4 expression was 55.0 percent. The percentage of cells stained by p16INK4 ranged from 10 to 100 percent, both in the group consisting of cervical intraepithelial neoplasia (CIN)1/HPV specimens and in the group of CIN2/CIN3 specimens with P value of 0.0001. p16INK4 expression was 48.3 percent in the CIN1/HPV group, as opposed to 94.3 percent in the CIN2/CIN3 group (P = 0.001), showing a statistically significant difference between the two groups. The quantitative method used here is simple and less subjective than the different semiquantitative methods described in the literature. In view of the different definitions of a p16INK4-positive case, it is almost impossible to compare the findings reported by different investigators. This study confirms the association between p16INK4 and CIN2 and CIN3 lesions. Moreover, it shows that some low grade lesions expressed high levels of this protein. This may indicate that such low grade lesions may be predisposed to progress to high grade lesions. This means that p16INK4 may be a strong marker for "neoplastic lesions" induced by HPV and not just an infection marker.

Human papillomavirus in oral lesions / Virus papiloma humano en lesiones orales

Growing evidence suggests a role for human papillomavirus (HPV) in oral cancer; however its involvement is still controversial. This study evaluates the frequency of HPV DNA in a variety of oral lesions in patients from Argentina. A total of 77 oral tissue samples from 66 patients were selected (cases); the clinical-histopathological diagnoses corresponded to: 11 HPV- associated benign lesions, 8 non-HPV associated benign lesions, 33 premalignant lesions and 25 cancers. Sixty exfoliated cell samples from normal oral mucosa were used as controls. HPV detection and typing were performed by polymerase chain reaction (PCR) using primers MY09, 11, combined with RFLP or alternatively PCR using primers GP5+, 6+ combined with dot blot hybridization. HPV was detected in 91.0% of HPV- associated benign lesions, 14.3% of non-HPV associated benign lesions, 51.5% of preneoplasias and 60.0% of cancers. No control sample tested HPV positive. In benign HPV- associated lesions, 30.0% of HPV positive samples harbored high-risk types, while in preneoplastic lesions the value rose to 59.9%. In cancer lesions, HPV detection in verrucous carcinoma was 88.9% and in squamous cell carcinoma 43.8%, with high-risk type rates of 75.5% and 85.6%, respectively. The high HPV frequency detected in preneoplastic and neoplastic lesions supports an HPV etiological role in at least a subset of oral cancers.

Crecientes evidencias sugieren que el virus Papiloma humano (HPV) tiene un rol en el cáncer oral; sin embargo su participación es todavía controvertida. Este estudio evalúa la frecuencia de ADN de HPV en una variedad de lesiones orales de pacientes de Argentina. Se seleccionaron 77 muestras de tejido oral de 66 pacientes (casos); el diagnóstico histo-patológico correspondió a: 11 lesiones benignas asociadas a HPV, 8 lesiones benignas no asociadas a HPV, 33 lesiones premalignas y 25 cánceres. Como controles se usaron 60 muestras de células exfoliadas de mucosa oral normal. La detección y tipificación de HPV se realizó por PCR empleando los primers MY09,11, seguida de RFLP, o PCR usando los primers GP5+, 6+ seguida de hibridación en dot blot. HPV fue detectado en 91% de las lesiones benignas asociadas a HPV, 14.3% de las lesiones benignas no asociadas, 51.5% de preneoplasias y 60% de cánceres. Ninguna muestra control resultó HPV positiva. En las lesiones benignas, 30% de las muestras HPV positivas correspondieron a tipos de alto riesgo, mientras que en las lesiones preneoplásicas la positividad ascendió a 59.9%. En cánceres, la detección de HPV en carcinomas verrugosos fue 88.9% y en carcinomas escamosos 43.8%, con 75.5% y 85.6% de tipos virales de alto riesgo, respectivamente. La alta frecuencia de HPV detectada en lesiones preneoplásicas y cánceres apoya un rol etiológico del HPV en, al menos, un subgrupo de cánceres orales.

Detección y tipificación de virus papiloma humano en lesiones preneoplásicas del cuello uterino mediante PCR-RFLP / Detection and typification of human papilloma virus in pre cancerous cervical lesions

Background: The association of different genotypes of human papilloma virus (HPV) with cervical cancer is well known. However, there is little information about their association with pre-cancerous lesions. Aim: To assess the frequency of different HPV genotypes in pre cancerous cervical lesions. Material and methods: A cervical sample was obtained by cytobrush in 15 women with low grade lesions and 40 women with high grade lesions, subjected to conization by loop electrical excision procedure (LEEP). Detection and typification of HPV was done by polymerase chain reaction and restriction fragment length polymorphism. Results: All women were infected with HPV. Eighty five percent of samples were typified. A unique HPV subtype was found in 76 percent of women. Fourteen percent had an infection with multiple subtypes and in 10 percent, the viral genotype was not identified. The most common subtypes found were HPV 16, HPV 52 and HPV 53. Conclusions: There is a high rate of infection with HPV with a high oncogenic risk among these women.

Inestabilidad microsatelital en lesiones preneoplásicas y neoplásicas del cuello uterino: Correlación con el genotipo del virus papiloma humano / Microsatellite instability and human papilloma virus genotypes in preneoplastic and neoplastic uterine cervix lesions

Background: The association between some specific human papilloma virus (HPV) types and cervix cancer is well known. However, the genetic conditions that favor the development of cervical cancer are less well known. Aim: To determine the presence of satellite instability (MSI) in preneoplastic and neoplastic lesions of the cervix and correlate these findings with HPV genotypes. Material and methods: Biopsy samples of cervical lesions were studied. Sixteen had low grade lesions, 22 had high grade lesions and 28 had an epidermoid cancer. Viral types were identified with polymerase chain reaction, dot-blot hybridization and restriction fragment length polymorphism. MSI was determined using a panel of eight highly informative microsatellites. Results: Microsatellite instability in at least one locus was observed in 91, 56 and 69 percent of low grade lesions, high grade lesions and epidermoid carcinomas, respectively. MSI-High grade, MSI-Low grade instability and microsatellite stability were observed in 5, 60 and 46 percent of samples, respectively. Two of three samples with high grade instability had HPV 52 genotype. Other viral subtypes had frequencies that ranged from 78 percent to 100 percent, with the exception of HPV16 that was present in only 53 percent of samples with low grade instability. Conclusions: Two thirds of biopsy samples from cervical lesions had MSI, mechanism that can be involved in the first stages of cervical carcinogenesis. The low frequency of high grade instability, its association with HPV52 and the low frequency of HPV16 in samples with low grade instability, suggest different coadjutant mechanisms in cervical carcinogenesis

Citologia anal para rastreamento de lesões pré-neoplásicas / Anal cytology for screening of pre-neoplasic lesions

OBJETIVO: Avaliar se a citologia anal com escova pode servir para rastreamento das lesões clínicas e subclínicas provocadas pelo HPV. MÉTODOS: Colhemos citologia anal, com escova, do canal anal de 102 doentes HIV-positivo com queixas proctológicas. Eram 86 homens e 16 mulheres com média etária de 37 anos. Destes, 33 negavam infecção pelo HPV, 14 haviam tratado verrugas, 28 tinham condilomas externos, sete apresentavam lesões internas e 20 os tinham em associação. O material foi enviado para exame de papanicolaou e coloração pela hematoxilina-eosina. Avaliamos as contagens de linfócitos T CD4+ para observar se o estado imunológico determinou as displasias mais avançadas. RESULTADOS: Somente um exame não pôde ser aproveitado. Os demais revelaram padrões celulares que variaram da normalidade até NIAa, incluindo a presença do HPV. Ocorreram 30 NIAs de baixo e 13 de alto grau em todos os grupos de doentes, com ou sem infecção pelo HPV. Em um dos doentes com NIAa e sem história prévia de infecção pelo HPV, e com úlcera no canal anal, a biópsia revelou carcinoma espinocelular invasivo. As médias de células T CD4+ nos portadores de NIA de baixo grau foi 281/mm³ e naqueles com NIAa foi 438/mm³. A análise estatística mostrou diferença significante, revelando que, ao contrário do esperado, displasias menos acentuadas acometem doentes com contagens menores de linfócitos T CD4+. Esse fato demonstra que a imunidade sistêmica isolada parece não interferir na gênese dessas lesões, sugerindo que aspectos da imunidade local devam ser estudados. A avaliação estatística feita com a tabela 2x2 revelou sensibilidade de 74 por cento e especificidade de 61 por cento. CONCLUSÃO: Acreditamos que a citologia anal possa servir para esse rastreamento, selecionando os doentes para colposcopia anal e biópsias.

BACKGROUND: High grade intra-epithelial neoplasias (HAIN) are probable precursors of anal carcinoma, with association to high-risk types of Human Papillomavirus (HPV). This progression could be related to severity of the dysplasia and, albeit not yet confirmed, treatment of these lesions would prevent the evolution to cancer. Standardization and improvement of screening methods should therefore be essential to treat or prevent precursor lesions, mainly in patients at risk such as seropositives to Human Immunodeficiency Virus (HIV). The aim of this study was to evaluate if anal cytology, with a cytobrush, could be useful to screen clinic and pre-clinic lesions provoked by HPV. METHODS: Brushes were used to obtain smears from the anal canal of 102 HIV-positive patients with proctologic complaints. There were 86 males and 16 females with a mean age of 37 years. HPV infection was denied by 33 patients, 14 had treated anal warts in the past, 28 had condylomas in the anal verge, seven had internal clinical lesions and 20 had both internal and external condylomas. The smears were submitted to Pappanicolaou and hematoxilin-eosin stains to identify cytological changes including HAIN. T CD4+ lymphocyte counts were also evaluated to check if the immunologic status caused more advanced dysplasia. RESULTS: One smear only proved insufficient. All the others revealed cellular patterns varying from normality to HAIN. Low grade AIN (LAIN) occurred in 30 and HAIN in 13 patients. One patient with HAIN, without a history of HPV infection in the past, presented an anal canal ulcer which at biopsy was diagnosed as invasive squamous-cell carcinoma. T CD4+ cells averaged 281/mm³ for LAIN patients and 438/mm³ for HAIN patients. Analyses disclosed a statistical difference, showing that despite expectations, more advanced dysplasias occurred in patients with higher counts of T CD4+ cells. This fact demonstrated that isolated systemic immunity did not...

Epidemiología de la infección cervical por virus Papiloma humano en Ushuaia: Argentina / Cervical infection epidemiology of human papillomavirus in Ushuaia: Argentina

El virus Papiloma humano (HPV) es un factor necesario para el desarrollo del cáncer cervical. El objetivo del estudio fue conocer la epidemiología de dicha infección en Ushuaia, Provincia de Tierra del Fuego, Argentina. Se realizó un estudio de caso-control de 132 cepillados endocervicales. La detección y tipificación del genoma viral fue realizada por la reacción en cadena de la polimerasa, con posterior análisis de polimorfismos de fragmentos de restricción o hibridación. La prevalencia general de la infección fue 41%, correspondiendo 26% a los controles y 71% a los casos. El grupo etario con mayor prevalencia de HPV fue el de 14 a 24 años. Los tipos virales más frecuentes en la población infectada fueron HPV16 (23%), HPV18 (11%) y HPV33/35 (8% cada uno), resultando infectados con tipos virales de alto riesgo el 30% de las muestras, 16% de los controles y 60% de los casos. El trabajo aporta los primeros datos sobre los tipos virales predominantes en Ushuaia. Los resultados demostraron una prevalencia menor que en regiones con alta incidencia de cáncer cervical, siendo el HPV16 el más frecuente. La información obtenida permitiría estimar la efectividad de las vacunas en vías de aprobación, en la población estudiada.

Genital infection with human papillomavirus (HPV) is decisive in the causation of cervical cancer. In order to evaluate the epidemiology of HPV infection in Ushuaia, Province of Tierra del Fuego, Argentina, 132 endocervical cytobrushes from preneoplastic and neoplastic cases and controls were studied. Detection and typing of the viral genome was performed by polymerase chain reaction, combined with a restriction fragment length polymorphism assay or hybridization. The overall prevalence of HPV infection was 41% in the population examined, with a frequency of 26% in the controls and 71% in the cases under study. The 14-24 age group showed the highest HPV prevalence. The most common viral types in the infected population were HPV 16 (23%), HPV 18 (11%), HPV 33 (8%) and HPV 35 (8%), while high risk viral types were detected in 30% of the samples, 16% of the controls and 60% of the cases. This study provides the first data on the predominant viral types in Ushuaia. Our results show lower levels of infection than in regions with a high incidence of cervical cancer, HPV 16 being the most prevalent viral type. This research may be useful for selecting a specific vaccine targeting the population examined.

Mecanismo de infección y transformación neoplásica producido por virus papiloma humano en el epitelio cervical / Infection mechanism and neoplastic transformation produced by human papilloma virus in cervical epithelium

Se presenta una revisión bibliográfica del mecanismo de infección y transformación neoplásica producida por el virus papiloma humano, de alto riesgo oncogénico, en el epitelio cervical. Se expone la interacción cápside receptor, internalización celular, expresión de genes tempranos, integración del genoma viral al de la célula huésped y algunos mecanismos vinculados a la proliferación y desarrollo neoplásico.

Utilidad en la combinación de oligonucleótidos universales para la detección del virus del papiloma humano en cáncer cervicouterino y lesiones premalignas / Usefulness of combining universal oligonucleotides in detecting human papillomavirus in cervical cancer and premalignant lesions

OBJETIVO: Determinar la frecuencia y distribución del virus del papiloma humano en los diferentes estadios que conforman la historia natural del cáncer cérvico uterino, y optimizar la detección mediante el uso de diferentes oligonucleótidos universales. MATERIAL Y MÉTODOS: Se trata de un estudio transversal, descriptivo, en el que las muestras fueron colectadas durante enero a diciembre de 1999. El procesamiento de las muestras y el análisis de los datos se realizaron en el Instituto Nacional de Cancerología en la Ciudad de México. Se hizo análisis comparativo con t de Student para valores continuos y con ji cuadrada para proporciones, y análisis de concordancia entre biopsia y exudado cervical con la prueba estadística de Kappa. Para la detección del virus se utilizó la técnica de la reacción en cadena de la polimerasa (PCR) con oligonucleótidos universales los cuales reconocen diferentes regiones del gen L1 (MY09/11; GP5/6; L1C1/2), y oligonucleótidos específicos para el VPH 16 y el VPH 18, así como secuenciación directa de los productos de la PCR. RESULTADOS: Se analizaron 154 muestras: 65 (42.2 por ciento) citologías normales, 45 (29.2 por ciento) lesiones de alto y bajo grado, y 44 (28.6 por ciento) de cáncer invasor. El VPH fue detectado en 95.5 por ciento de los casos de cáncer invasor, en 91.6 por ciento de lesiones de alto grado, en 66.7 por ciento de lesiones de bajo grado y en 23.1 por ciento de citologías normales, por la PCR con al menos uno de los juegos de oligonucleótidos utilizados. La detección fue más eficiente en las muestras obtenidas por biopsia que en los exudados cervicovaginales. El porcentaje total de detección del VPH con un juego de oligonucleótidos universales (37.6 por ciento) aumentó sustancialmente (60.4 por ciento) al combinarlo con otros dos juegos de oligonucleótidos universales. CONCLUSIONES: La presencia del VPH de alto riesgo es elevada inclusive en mujeres con epitelios cervicales con diagnóstico citológico normal. La detección del VPH mejora al utilizar distintos juegos de oligonucleótidos universales que reconocen la región L1 del VPH. Con una adecuada toma de muestra, el análisis para detección de ADN en exudado cérvico-vaginal es una buena alternativa de diagnóstico del VPH.