Association of human papillomavirus (HPV), p16, p53 and p63 expression with non-bilharzia-associated squamous cell carcinoma of the bladder and algorithm construction for histopathological grading prediction

ABSTRACT Objective To investigate the expression of human papillomavirus (HPV), p16, p53, and p63 in non-schistosomiasis-related squamous cell carcinoma of the bladder and to develop an accurate and automated tool to predict histological classification based on clinicopathological features. Methods Twenty-eight patients with primary bladder pure squamous cell carcinoma who underwent cystectomy or transurethral resection of bladder tumor (TURBT) for bladder cancer between January 2011 and July 2017 were evaluated. Clinical data and follow-up information were obtained from medical records. Formalin-fixed, paraffin-embedded surgical specimens were used for immunohistochemical staining for p16, p53, and p63. Human papillomavirus detection was evaluated by PCR. Statistical analysis was performed, and statistical significance was set at p<0.05. Finally, decision trees were built to classify patients' prognostic features. Leave-one-out cross-validation was used to test the generalizability of the model. Results Neither direct HPV detection nor its indirect marker (p16 protein) was identified in most cases. The absence of p16 was correlated with less aggressive histological grading (p=0.040). The positive p16 staining detection found only in pT1 and pT2 cases in our sample suggests a possible role for this tumor suppressor protein in the initial stages of bladder squamous cell carcinoma. The decision trees constructed described the relationship between clinical features, such as hematuria/dysuria, the level of tumor invasion, HPV status, lymphovascular invasion, gender, age, compromised lymph nodes, and tumor degree differentiation, with high classification accuracy. Conclusion The algorithm classifier approach established decision pathways for semi-automatic tumor histological classification, laying the foundation for tailored semi-automated decision support systems for pathologists.

Genotipos del Virus del Papiloma Humano en citología alterada del cuello uterino / Human Papillomavirus genotypes in altered cervical cytology / Genótipos do papilomavírus humano em citologia cervical alterada

En Ecuador el cáncer de cuello uterino se ubica en el segundo lugar y, está relacionada a una infección genital persistente por el virus del papiloma humano (VPH) de alto riesgo. Objetivo general: determinar la relación entre virus del papiloma humano de alto riesgo y las lesiones intraepiteliales del cuello uterino, en mujeres de 21 a 65 años en tres cantones de la provincia de El Oro, periodo 2019. Se trata de un estudio de tipo descriptivo relacional de corte transversal. Se realizó el estudio en 109 mujeres que cumplieron los criterios de inclusión. Para la relación de las variables se utilizó los estadísticos del Chi cuadrado (con valor de p 0,05) y, la lesión más frecuente fue el de células escamosas atípicas de importancia no determinada. Se concluye que las lesiones intraepiteliales fueron más frecuentes que las reportadas en la literatura como general, y los genotipos 39, 16, 18 estuvieron presentes en las lesiones intraepiteliales de bajo grado del cuello uterino.

In Ecuador, cervical cancer is in second place and is related to persistent genital infection by high-risk human papillomavirus (HPV). General objective: to determine the relationship between high-risk human papillomavirus and cervical intraepithelial lesions in women aged 21 to 65 years in three cantons of the province of El Oro, period 2019. This is a descriptive relational cross-sectional study. The study was conducted in 109 women who met the inclusion criteria. Chi-square statistics (with p value 0.05) and the most frequent lesion was atypical squamous cell of undetermined significance. It is concluded that intraepithelial lesions were more frequent than those reported in the literature in general, and genotypes 39, 16, 18 were present in low-grade intraepithelial lesions of the cervix.

O câncer cervical é a segunda principal causa de câncer cervical no Equador e está relacionado à infecção genital persistente por papilomavírus humano de alto risco (HPV). Objetivo geral: determinar a relação entre papilomavírus humano de alto risco e lesões intra-epiteliais do colo uterino em mulheres de 21 a 65 anos de idade em três cantões da província de El Oro, período 2019. Este é um estudo descritivo, transversal e relacional. O estudo foi conduzido em 109 mulheres que preenchiam os critérios de inclusão. As estatísticas qui-quadradas (com valor p 0,05) e a lesão mais freqüente foi a célula escamosa atípica de importância indeterminada. Conclui-se que as lesões intra-epiteliais eram mais freqüentes do que as relatadas na literatura em geral, e os genótipos 39, 16, 18 estavam presentes em lesões intra-epiteliais de baixo grau do colo uterino.

Relationship between Papillomavirus vaccine, vaginal microbiome, and local cytokine response: an exploratory research.

INTRODUCTION: The influence of vaccination on composition of the human microbiome at distinct sites has been recognized as an essential component in the development of new vaccine strategies. The HPV vaccine is widely used to prevent cervical cancer; however, the influence of HPV vaccine on the vaginal microbiota has not been previously investigated. In his study, we performed an initial characterization of the microbiome and cytokine composition in the vagina following administration of the bivalent vaccine against HPV 16/18. MATERIAL AND METHODS: In this exploratory study, fifteen women between 18 and 40 years received three doses of the HPV-16/18 AS04-adjuvanted vaccine (Cervarix®). Cervicovaginal samples were collected before the first dose and 30 days after the third dose. HPV genotyping was performed by the XGEN Flow Chip technique. The cytokines IFN-γ, IL-2, IL-12p70, TNF-α, GM-CSF, IL-4, IL-5, IL-10, and IL-13 were quantitated by multiplex immunoassay. The vaginal microbiome was identified by analysis of the V3/V4 region of the bacterial 16S rRNA gene. RESULTS: The most abundant bacterial species in the vaginal microbiome was Lactobacillus crispatus, followed by L. iners. Bacterial diversity and dominant organisms were unchanged following vaccination. Small decreases in levels of pro and anti-inflammatory cytokines were observed following HPV vaccination, but there was no association between vaginal cytokine levels and microbiome composition. CONCLUSION: Vaginal microbiome is not altered following administration of the standard three-dose HPV-16/18 AS04-adjuvanted (Cervarix®) vaccine.

E6/E7 Variants of Human Papillomavirus 16 Associated with Cervical Carcinoma in Women in Southern Mexico.

Persistent infection with the human papillomavirus 16 (HPV 16) is the cause of half of all cervical carcinomas (CC) cases. Moreover, mutations in the oncoproteins E6 and E7 are associated with CC development. In this study, E6/E7 variants circulating in southern Mexico and their association with CC and its precursor lesions were evaluated. In total, 190 DNA samples were obtained from scrapes and cervical biopsies of women with HPV 16 out of which 61 are from patients with CC, 6 from patients with high-grade squamous intraepithelial lesions (HSIL), 68 from patients with low-grade squamous intraepithelial lesions (LSIL), and 55 from patients without intraepithelial lesions. For all E7 variants found, the E7-C732/C789/G795 variant (with three silent mutations) was associated with the highest risk of CC (odd ratio (OR) = 3.79, 95% confidence interval (CI) = 1.46-9.85). The analysis of E6/E7 bicistron conferred to AA-a*E7-C732/C789/G795 variants revealed the greatest increased risk of CC (OR = 110, 95% CI = 6.04-2001.3), followed by AA-c*E7-C732/C789/G795 and A176/G350*E7-p. These results highlight the importance of analyzing the combinations of E6/E7 variants in HPV 16 infection and suggest that AA-a*E7-C732/C789/G795, AA-c*E7-C732/C789/G795, and A176/G350*E7-p can be useful markers for predicting CC development.

In silico analysis of genomic variables associated to HPV16 integration sites / Análisis in silico de variables genómicas asociadas a sitios de integración de HPV16

Background: Despite current prophylactic interventions, a significant proportion of patients suffers a cancer-specific mortality, leading to a global awareness of the importance of identifying factors associated to the etiology of HPV-associated cancer. According to this, HPV-DNA integration into human genome is an important event in the pathogenesis. Purpose: To identify in silico, molecular regions of the genome where the HPV integration events occur Methods: We performed a bioinformatic study based on a systematic search in Medline through PubMed, Embase and Lilacs from inception to April 2019. We used the UCSC Genome Browser Home (https://genome.ucsc.edu) to evaluate the genetic environment. Results: HPV integration sites by anatomical location related to cervical cancer were 374 (61%). In addition, 325 (87%) of these integration sites had HPV-16, 21 (5%) had HPV-18 and 28 (7%) had another type of genotype. Oro-pharyngeal cavity was the second anatomic site with 162 (26%) integration sites. It is noteworthy that the HPV-16 was found integrated into 160 (99%) analyzed sites. Conclusion: Our results suggest that many of the integration sites reported in the scientific literature are HPV 16 from squamous cell carcinomas and 50% of HPV16 were integrated into transcriptional units that might affect the expression of gene target.

Antecedentes: A pesar de las intervenciones profilácticas actuales, una proporción significativa de pacientes muere debido al cáncer, lo que aumenta la conciencia global de la importancia de identificar los factores asociados a la etiología del cáncer asociado al VPH. Según esto, la integración del ADN-VPH en el genoma humano es un evento importante en la patogénesis. Propósito: Identificar in silico, las regiones moleculares del genoma donde ocurren los eventos de integración del VPH Métodos: Realizamos un estudio bioinformático basado en una búsqueda sistemática en Medline a través de PubMed, Embase y Lilacs desde el inicio hasta abril de 2019. Utilizamos el UCSC Genome Browser Home (https://genome.ucsc.edu) para evaluar el entorno genético. Resultados: Los sitios de integración del VPH relacionados con el cáncer de cuello uterino fueron 374 (61%). Además, 325 (87%) de estos sitios de integración tenían VPH-16, 21 (5%) tenían VPH-18 y 28 (7%) tenían otro tipo de genotipo. La cavidad orofaríngea fue el segundo sitio anatómico con 162 (26%) sitios de integración. Es de destacar que el VPH-16 se encontró integrado en 160 (99%) sitios analizados. Conclusión: Nuestros resultados sugieren que muchos de los sitios de integración reportados en la literatura científica que presentan al VPH-16 son carcinomas de células escamosas y que el 50% de estos VPH-16 se integraron en unidades transcripcionales que podrían afectar la expresión de algún gen objetivo.

Prevalencia de los Genotipos del Virus del Papiloma Humano en mujeres de 25 a 65 años / Prevalence of Human Papillomavirus Genotypes in women aged 25 to 65 years

Introducción: La infección que ocasiona el Virus del Papiloma Humano (VPH), tiene alta prevalencia en mujeres sexualmente activas. Generalmente es pasajera, pero al existir algunos factores relacionados pueden llegar a desarrollar cáncer cervicouterino. Dado que la enfermedad se desarrolla con lentitud la detección en etapas tempranas ha permitido poner en evidencia la presencia del virus en las células antes que puedan transformarse y volverse tumorigénicas. El objetivo de este estudio fue establecer la prevalencia de los genotipos del Virus del Papiloma Humano en mujeres de 25 a 65 años en un grupo de pacientes de un centro oncológico en Cuenca 2017 ­2018. Métodos:Es un estudio descriptivo, retrospectivo, analítico, en el cual se recopiló información de las historias clínicas y registros físicos del Laboratorio de Biología Molecular y del sistema médico de SOLCA -Cuenca, SOFTCASE, para establecer la prevalencia de VPH durante el periodo 2017 -2018.Se utiliza ODDS Ratio para demostrar asociación entre las variables demográficas y los grupos de serología de VPH de riesgo alto versus VPH De riesgo bajo. Resultados:Se incluyeron 594casos, con edad entre36 y 40 años n=103/594 (17.3%). De estado civil casadas n=318/594 (53.5%). Con paridad igual a 2 n=159/594 (26.8%). Casospositivos de VPH fueron 424/594 (71.38%) IC95% (71.23% a 71.53%), Genotipos de alto riesgo con el 58.01%, genotipos de probable bajo riesgo con el 33.25% y genotipos de bajo riesgo 8.72%. La prevalencia del 50% de la población positiva según el genotipo lo explicalos VPH 16, 71, 58, 6 y 31. De este grupo los VPH con serología 16, 58 y 31 tienen un riesgo Alto de malignidad. No se reportó asociación entre los VPH de alto riesgo con alguna de las variables demográficas. Conclusión:El grupo etario con mayor número de casos positivos perteneció a las mujeres de entre 36 y 40 años de edad, con paridad igual a 2 y de estado civil casadas. El subtipo VPH-16 fue el genotipo más prevalente del grupo de alto riesgo de malignidad. El subtipo VPH-71 fue el segundo genotipo más prevalente con un perfil de probable bajo riesgo de malignidad.

AbstractIntroduction:The infection caused by the Human Papilloma Virus (HPV) has a high prevalence in sexually active women. It is generally temporary, but when there are some related factors, they can develop cervical cancer. Since the disease develops slowly, detection in early stages has made it possible to reveal the presence of the virus in cells before they can transform and become tumorigenic. The objective of this study was to establish the prevalence of Human Papilloma Virus genotypes in women aged 25 to 65 years in a group of patients from an oncology center in Cuenca 2017-2018. Methods: It is a descriptive, retrospective, analytical study, in which information was collected from the medical records and physical records of the Molecular Biology Laboratory and the SOLCA -Cuenca medical system, SOFTCASE, to establish the prevalence of HPV during the period 2017 -2018. ODDS Ratio is used to demonstrate association between demographic variables and high-risk HPV versus low-risk HPV serology groups. Results: 594 cases were included, aged between 36 and 40 years, n = 103/594 (17.3%). Marital status married n = 318/594 (53.5%). With parity equal to 2 n = 159/594 (26.8%). Positive HPV cases were 424/594 (71.38%) 95% CI (71.23% to 71.53%), high risk genotypes with 58.01%, probable low risk genotypes with 33.25% and low risk genotypes 8.72%. The prevalence of 50% of the positive population according to genotype is explained by HPV 16, 71, 58, 6 and 31. Of this group, HPV with serology 16, 58 and 31 have a high risk of malignancy. No association was reported between high-risk HPV with any of the demographic variables. Conclusion: The age group with the highest number of positive cases belonged to women between 36 and 40 years of age, with parity equal to 2 and married marital status. The HPV-16 subtype was the most prevalent genotype in the group at high risk of malignancy. The HPV-71 subtype was the second most prevalent genotype with a profile of probable low risk of malignancy.

Frequency of high-risk genotypes of human papilloma virus in oral lesions / Frecuencia de genotipos alto riesgo del virus papiloma humano en lesiones bucales

Some genotypes of the human papilloma virus (HPV) in the oral cavity cause genetic instability that may lead to cancer. Clinical and histological diagnoses are key tools; however, molecular techniques allow predicting, detecting and monitoring the disease. Objective: To identify the frequency of four high-risk HPV genotypes and their association with lesions in the oral cavity. Materials and Methods: Descriptive cross-sectional study with a sample of 48 patients diagnosed with hyperplastic lesions and others currently classified as potentially malignant disorders (PMDs) of the oral cavity, who underwent biopsies, histopathological analysis, and HPV16, 18, 31, and 45 detection and genotyping by polymerase chain reaction (PCR). Results: Epithelial hyperplasia was the most frequent lesion found in 45.8% (n=22) of patients. Nicotine palatinus and leukoplakia were found in 8.3% and 6.2%, respectively; oral cancer in 6.2%. The total frequency of HPV was 12.5% (6/48). Oral papilloma was found in 6.1% (3/48), and nicotine palatinus and oral cancer in 2.0% each (1/48). HPV16, HPV31, and HPV45 were detected, while HPV18 was not observed. HPV16 was the most frequent genotype found (4 out of 6 patients), while HPV31 and HPV45 were found in one patient each. Only one genotype per lesion was found. The presence of HPV was associated with lesions (χ2=11.810; p=0.0375). No significant association with age and gender was found. Conclusion: High-risk HPV continues to be present in oral lesions. The HPV16 viral genotype was the most frequent in the studied lesions.

Algunos genotipos del virus del papiloma (VPH) en boca, producen inestabilidad genética dando lugar al cáncer. El diagnóstico clínico e histológico son herramientas claves, sin embargo, técnicas moleculares permiten predecir, detectar y dar seguimiento a la enfermedad. Objetivo: Identificar la frecuencia de cuatro genotipos del VPH de alto riesgo y su asociación con lesiones en cavidad bucal. Material y Métodos: Estudio descriptivo de corte transversal con una muestra de 48 pacientes diagnosticados con lesiones hiperplásicas y otros clasificados actualmente como desordenes potencialmente malignos (DPM) de la cavidad bucal, a quienes se les realizó biopsias, análisis histopatológico y detección y genotipificación VPH16, 18, 31, y 45 mediante reacción en cadena a la polimerasa (PCR). Resultado: La hiperplasia epitelial fue la lesión más frecuente en 45,8% (n=22). La palatinitis nicotínica y la leucoplasia, se encontraron 8,3% y 6,2% respectivamente, cáncer oral, en 6,2%. La frecuencia total de VPH fue 12,5% (6/48). El papiloma oral estuvo en un 6,1% (3/48), palatinitis nicotínica y cáncer oral en 2,0% (1/48).Se detectó VPH16, VPH31 y VPH45, mientras que VPH18 estuvo ausente. ElVPH16 fue el de mayor frecuencia con 66,7% (4/6), el VPH31 y VPH45 se encontraron en 16,7% (1/6). No se evidenció más de un genotipo por lesión. La presencia de VPH estuvo asociado con las lesiones (χ2=11,810; p=0,0375). No se encontró asociación significativa con edad y género. Conclusión: El VPH de alto riesgo sigue estando presente en lesiones bucales. El genotipo viral VPH16 se encontró con mayor frecuencia en las lesiones estudiadas.

Indoleamine 2,3-dioxygenase and tryptophan 2,3-dioxygenase expression in HPV infection, SILs, and cervical cancer.

BACKGROUND: Human papillomavirus (HPV) infection is the central factor for cervical cancer, whereas epithelial immune mechanisms contribute to the progression of HPV infection and its associated lesions. The authors evaluated the expression of indoleamine 2,3-dioxygenase (IDO) and tryptophan 2,3-dioxygenase (TDO) in cervicovaginal samples from women with normal cervical epithelium or with different degrees of squamous intraepithelial lesions (SILs) and cervical cancer. METHODS: IDO expression was analyzed by immunocytochemistry in liquid-based cytology samples from 165 women, of whom 42 had cervical changes subclassified as low-grade SIL (n = 6), high-grade SIL (n = 30), or squamous cell carcinoma (SCC) (n = 6), and 123 had negative Papanicolaou smears. IDO and TDO expression also were analyzed by immunohistochemistry, and HPV and other genital pathogens were evaluated by polymerase chain reaction analysis. RESULTS: Low IDO expression was observed in normal cervical epithelium irrespective of HPV status. Increased numbers of IDO-positive squamous cells and IDO-positive leukocytes were observed in women with SIL or SCC. TDO expression was detected in leukocytes infiltrating the stroma around intraepithelial or invasive cervical lesions. Higher IDO levels were detected in organotypic epithelial cultures established from keratinocytes transduced with the HPV16 E6/E7 oncoproteins. CONCLUSIONS: The upregulation of IDO expression in leukocytes and squamous cells in HPV-associated SIL and SCC suggests that immunosuppressive mechanisms involving tryptophan metabolism may have a role in cervical carcinogenesis. Although previous studies have suggested the role of IDO in HPV pathogenesis, this is the first evidence of TDO involvement in the process. Furthermore, the current data emphasize the role of leukocytes, especially neutrophil-like cells, as an IDO source.

Integration, Viral Charge and E2 mRNA Levels in the Progresion of Intraepitelial Cervical Lesions / Integración, carga viral y niveles de arn mensajero de e2 de vph 16 en la progresión de lesiones intraepiteliales cervicales

RESUMEN Entre las lesiones intraepiteliales escamosas cervicales (LIE) es importante distinguir aquellas asociadas con mayor riesgo de cáncer de cuello uterino. El objetivo de este trabajo fue evaluar si los niveles de expresión de E2 del VPH16 en mujeres con LIE y con evidencia de integración viral se asocian con el grado de la lesión. Se analizaron 109 cepillados cervicales positivos para VPH 16 provenientes de 19 mujeres sin LIE, 45 mujeres con LIE de bajo grado (LIEBG) y 45 mujeres con LIE de alto grado (LIEAG). Se cuantificó el número de copias de ARNm de E2 y de los genes E2 y E6 mediante PCR en tiempo real para determinar la carga viral (E6) y la proporción E2/E6 para evaluar la integración viral. Se encontraron frecuencias similares de expresión de E2 en LEIBG y LEIAG 15/45 (33 %), la frecuencia en mujeres sin lesión fue menor 3/19 (15,8 %), todos los casos en los que se observó expresión del gen E2 tenían mezcla de ADN viral episomal e integrado. La carga viral aumentó significativamente a mayor grado de la lesión (ρ =0,049), mientras que la proporción E2/E6 disminuyó (ρ=0,049). El análisis ROC mostró una baja capacidad de los tres parámetros virales para distinguir entre lesiones de bajo y alto grado. En conclusión, aunque las lesiones con presencia de ADN viral mixto e integrado y expresión de E2 podrían estar en menor riesgo de progresión, y la carga viral y la integración se relacionaron con mayor gravedad de la lesión, su valor clínico como biomarcadores de LEIAG es limitado.

ABSTRACT It is important to distinguish among squamous intraepithelial lesions (SIL) those associated with increased risk cervical cancer. Our aim was to evaluate if the expression level of gen E2 in women with SIL and evidence of viral integration is associated to the grade of lesion. Cervical scrapes HPV16 positive from 19 women with normal histology, 45 women with low-grade SIL (LSIL) and 45 women with high-grade SIL (HSIL) were analyzed. Real-time PCR was used to quantify the mRNA of E2 and E2 and E6 genes to calculate viral load (E6) and the ratio E2/E6 to assess viral integration. Similar frequencies of E2 expression were found in LSIL and HSIL15/45 (33 %), the frequency in women without SIL was lower 3/19 (15.8 %), and all cases with E2 gene expression had mixed episomal and integrated viral DNA. The viral load increased significantly with the grade of SIL (ρ= 0.049), while E2/E6 ratio decreased (ρ=0.049). The ROC analysis showed low capacity of the three viral parameters analyzed to distinguish between low and high grade SIL. In conclusion although SIL with mixed and integrated viral DNA with E2 expression could be at lower risk of progression, and viral load and integration were associated with higher severity of the lesion, its clinical value as biomarkers of HSIL is limited.

Methylation of the L1 gene and integration of human papillomavirus 16 and 18 in cervical carcinoma and premalignant lesions.

High-risk human papillomavirus (HPV) is the primary cause of cervical carcinoma (CC). Viral integration into the host chromosomes is associated with neoplastic progression, and epigenetic changes may occur as a result. The objective of the present study was to analyze HPV L1 gene methylation and to compare the use of quantitative polymerase chain reaction (qPCR), in situ hybridization (ISH) and L1 methylation analysis as methods for detecting HPV integration. Cervical scrapes or biopsy samples positive for HPV 16 or 18, from 187 female patients with CC, squamous intraepithelial lesions (SILs) or no intraepithelial lesion (non-IL) were analyzed. Methylation of the L1 gene was determined using bisulfite modification followed by PCR, and HPV integration was subsequently analyzed. HPV 16 L1 gene methylation was revealed to increase with histological grade, with statistically significant differences observed as follows: Low-grade SIL vs. CC, P<0.0001 and non-IL vs. CC, P<0.0001. HPV 18 L1 gene methylation also increased according to histological grade, however, no statistically significant differences were observed. Methylation at CpG site 5608 of the HPV 16 L1 gene was associated with all grades of cervical lesions, whereas methylation at CpG site 5617 demonstrated the strongest association with CC (odds ratio, 42.5; 95% confidence interval, 4.7-1861; P<0.0001). The concordance rates between the various methods for the detection of the physical status of HPV 16 and HPV 18 were 96.1% for qPCR and ISH, 76.7% for qPCR and L1 gene methylation, and 84.8% for ISH and L1 gene methylation. In conclusion, methylation of the HPV 16 L1 gene increases significantly according to the grade of the cervical lesion, and methylation at CpG sites 5608 and 5617 of this gene may be used as prognostic biomarkers. ISH and L1 gene methylation have good concordance with qPCR with regards to the detection of HPV integration. Therefore, these are useful methods in determining the physical state of HPV.

Pigmented Bowen's disease associated with high-risk HPV simulating melanoma of the hand

Abstract: Bowen's disease is an in situ squamous cell carcinoma of the skin with only 2% of pigmented cases reported. It is clinically characterized by papules and plaques of blackened surface that may be caused either by sun damage - usually in photoexposed areas in elderly individuals - or by human papillomavirus infection - usually in the anogenital region of young adults. Dermoscopic aspects of Bowen's disease are discussed for over a decade, but with no definitive criteria that would lead to a definitive diagnosis. We present a case of Bowen's disease affecting the finger of a 57-year-old Asian patient. The lesion clinically and dermoscopically simulated a melanoma. Histopathological findings suggested the diagnosis of pigmented Bowen's disease. Pigmented Bowen's disease should be considered a differential diagnosis of melanoma, since its clinical and dermoscopic criteria are unspecific. Histopathological examination remains the gold standard for the diagnosis of the disease.

Prevalence of Human Papillomavirus Infection and Cervical Cancer Screening among Riverside Women of the Brazilian Amazon / Prevalência da infecção pelo papilomavírus humano e rastreamento do câncer em mulheres ribeirinhas da Amazônia brasileira

Abstract Purpose The aim of this study was to evaluate the overall and type-specific prevalence of human papillomavirus (HPV) infection among females living in riverside communities in the state of Pará, in the Eastern Brazilian Amazon. These communities are inhabited by low-income people, and are accessible only by small boats. Cervical cytology and risk factors for HPV infection were also assessed. Methods Cervical samples from 353 women of selected communities were collected both for Papanicolau (Pap) test and HPV detection. Conventional polymerase chain reaction (PCR) and real-time PCR were used to assess the overall and type-specific prevalence of HPV-16 and HPV-18, the main oncogenic types worldwide. Epidemiological questionnaires were used for the assessment of the risk factors for HPV infection. Results The mean age of the participants was 37 years (standard deviation [SD] ± 13.7). Most were married or with a fixed sexual partner (79%), and had a low educational level (80%) and family monthly income (< U$ 250; 53%). Overall, HPV prevalence was 16.4% (n = 58), with 8 cases of HPV-16 (2.3%) and 5 of HPV-18 (1.4%). Almost 70% of the women surveyed had never undergone the Pap test. Abnormal cytology results were found in 27.5% (n = 97) of the samples, with higher rates of HPV infection according to the severity of the lesions (p = 0.026). Conclusions The infections by HPV-16 and HPV-18 were not predominant in our study, despite the high prevalence of overall HPV infection. Nevertheless, the oncogenic potential of these types and the low coverage of the Pap test among women from riverside communities demonstrate a potential risk for the development of cervical lesions and their progression to cervical cancer, since the access to these communities is difficult and, in most cases, these women do not have access to primary care and public health services.

Resumo Objetivo O objetivo deste estudo foi avaliar a prevalência global e tipo-específica da infecção pelo papilomavírus humano (HPV) entre mulheres que vivem em comunidades ribeirinhas do estado do Pará, Amazônia oriental, Brasil. Estas comunidades são habitadas por pessoas de baixa renda, e são acessíveis somente por meio de pequenos barcos. A citologia cervical e os fatores de risco para a infecção por HPV também foram avaliados. Métodos Amostras cervicais de 353 mulheres de comunidades selecionadas foram coletadas para a análise citológica e para a detecção do HPV. A prevalência global e tipoespecífica dos HPV-16 e HPV-18, principais tipos oncogênicos no mundo, foram avaliadas por meio de reação em cadeia de polimerase (PCR) convencional e PCR em tempo real. Os fatores de risco para a infecção por HPV foram avaliados a partir de questionários epidemiológicos. Resultados A idade média das participantes foi de 37 anos (desvio padrão [DP] ± 13,7). A maioria era casada ou tinha um parceiro sexual fixo (79%) e baixo nível de escolaridade (80%) e de renda familiar mensal (< U$ 250; 53%). A prevalência global do HPV foi de 16,4% (n = 58), com 8 casos de HPV-16 (2,3%) e 5 casos de HPV-18 (1,4%). Aproximadamente 70% das mulheres entrevistadas nunca tinha realizado o exame preventivo de Papanicolau. Os resultados citológicos anormais foram encontrados em 27,5% (n = 97) das amostras, com taxas mais altas da infecção por HPV de acordo com a severidade das lesões (p = 0,026). Conclusões As infecções por HPV-16 e HPV-18 não foram predominantes em nosso estudo, apesar da alta prevalência global da infecção por HPV. No entanto, o potencial oncogênico desses tipos e a baixa cobertura do exame de Papanicolau entre mulheres de comunidades ribeirinhas demonstram um risco potencial para o desenvolvimento de lesões cervicais e sua progressão para o câncer de colo do útero, uma vez que o acesso a essas comunidades é difícil e, na maioria dos casos, estas mulheres não têm acesso aos serviços de atenção primária e de saúde pública.

Comparação da prevalência do HPV-16 em pacientes com carcinoma espinocelular de boca e lesões potencialmente malignas bucais / Comparison of the HPV-16 prevalence in patients with oral squamous cell carcinoma and oral potentially malignant disorders

O papilomavírus humano (HPV) subtipo 16 é um fator de risco para o desenvolvimento do carcinoma espinocelular (CEC) de orofaringe. No entanto, o papel do mesmo na carcinogênese oral, bem como a associação com as lesões potencialmente malignas, permanece controverso. O objetivo deste estudo foi comparar a prevalência do HPV-16, em amostras de tecido fresco, obtidas de 27 pacientes com CEC oral, 37 pacientes com leucoplasia bucal, 24 pacientes com líquen plano bucal (LPB) e 32 pacientes controle, correlacionando a presença do HPV com as variáveis clínico-patológicas em uma população da região noroeste do estado de São Paulo - Brasil. Realizouse a extração do DNA das amostras e a verificação da presença do HPV-16 por meio da Real-Time Polymerase Chain Reaction. Todas as amostras foram negativas para o HPV-16 nos quatro grupos estudados. Conclui-se que a ausência do HPV-16 nas amostras de CEC bucal, leucoplasia e LPB indica que a infecção pelo mesmo não é comum e não representa um fator de risco importante na carcinogênese oral na população da região noroeste do estado de São Paulo ­ Brasil(AU)

Human Papillomavirus (HPV), specially subtype 16, is a known risk factor for the oropharyngeal squamous cell carcinoma (SCC) development. However, HPV role in oral carcinogenesis, as well as in potentially malignant oral lesions remains controversial. The goal of the present study was to compare the HPV16 prevalence, in fresh tissue samples obtained from 27 oral SCC patients, 34 oral leukoplakia (OL) patients, 24 oral lichen planus (OLP) patients and 32 control patients, correlating HPV presence with the clinicopathological variables in a population from northwest region of the Sao Paulo state - Brazil. DNA extraction was carried out and all samples were submitted to Real-Time PCR for HPV-16 DNA detection. We found that all fresh tissue samples of oral SCC, OL, OLP and oral normal mucosa were negative for HPV-16. We conclude that HPV-16 absence in oral SCC, OL and OLP samples indicates that its infection is uncommon and does not represent an important risk factor in oral carcinogenesis in the population from northwest region of the Sao Paulo state ­ Brazil(AU)

Identificación molecular de genotipos papilomavirus humanos en pacientes con cáncer de cuello uterino / Molecular identification of human papilloma virus genotypes in patients with cervical cancer

Fundamento: la principal causa para el cáncer cervico uterino es el papilomavirus humano de alto riesgo. No existen antecedentes de estudios moleculares para la tipificación de papilomavirus humano en la población de Camagüey. La reacción en cadena de la polimerasa es una técnica de Biología Molecular que se ha usado desde siempre para el diagnóstico clínico; esta permite confirmar la presencia del ADN del Papilomavirus en el ADN total extraído a partir de muestras de pacientes con cáncer de cuello uterino. Objetivo: demostrar por primera vez los genotipos papilomavirus humano de alto riesgo circulantes, que causan cáncer de cuello uterino en la población femenina de Camagüey, Cuba. Métodos: se realizó un estudio analítico prospectivo donde se estudiaron 22 pacientes femeninas de la provincia de Camagüey, que fueron atendidas en la consulta de Patología de cuello del Hospital Ginecoobstétrico. La identificación y tipificación de los genotipos papilomavirus humano se realizó mediante el procedimiento molecular polimorfismo de longitud en los fragmentos de restricción. Resultados: el 63, 6 % de los pacientes presentaron lesiones tipo exofítica, el 4, 5 % endofítica y el 31, 8 % de otros tipos. Este estudio confirmó que los genotipos papilomavirus humanos de alto riesgo circulantes en la provincia Camagüey son los genotipos 16 y 31, donde el más frecuente fue el genotipo 16. Conclusiones: la presente investigación constituye el primer reporte de un estudio molecular de papilomavirus humanos a partir de muestras de pacientes con cáncer de cuello uterino en la provincia de Camagüey, Cuba. Estos resultados, junto a los obtenidos por otros autores, tienen una contribución importante en el diseño de preparados vacunales preventivos o terapéuticos, cada vez más efectivo hacia una solución anticipada para el cáncer de cuello uterino en Cuba.

Background: it is demonstrated that the main cause of cervical cancer is high risk humanp virus. There is no precedent of molecular studies for the typing of Human Papilloma Virus in the population of Camagüey. Polymerase chain reaction is Molecular Biology technique that has been used traditionally for the clinical diagnosis and other purposes. This technique allows confirming the presence of papillomavirus´DNA in the total extracted DNA, from samples of patients with cervical cancer. Objective: to demonstrate for the first time existing high-risk human papilloma virus genotypes that cause cervical cancer in female population of Camagüey, Cuba. Methods: a prospective analytic study was conducted, in which 22 female patients of the province of Camagüey were studied. They received medical attention at Ana Betancourt Hospital. Identification and typing of the Human Papilloma Virus genotypes was carried through the molecular procedure Restriction Fragment Length Polymorphism. Results: patients who presented exophytic lesions accounted for 63, 6%, 4, 5 % had endophytic type, and 31, 8 % presented other types. This study confirmed that high-risk Human Papilloma Virus genotypes existing in the province of Camagüey are genotypes 16 and 31, and the most frequent is 16. Conclusions: this research is the first report of a molecular study of Human Papilloma Virus from samples of patients with cervical cancer in the province of Camagüey, Cuba. These results, along with the ones obtained by other authors, make an important contribution in the design of the increasingly effective therapeutic and preventive vaccine to an anticipated solution to cervical cancer in Cuba.

Detection of human papillomavirus in dental biofilm and the uterine cervix of a pregnant adolescent / Detecção do papilomavírus humano em biofilme dental e no colo uterino de uma adolescente grávida

CONTEXT: Adolescence and pregnancy are considered to be risk factors for human papillomavirus (HPV) infection. The relationship between this infection in the uterine cervix and oral HPV infection is controversial. CASE REPORT: This report describes a case of a pregnant 16-year-old adolescent who presented HPV infection in the uterine cervix and the mouth. Smears were collected from the cervix and the tongue/palate. Dental biofilm samples were also collected. The microarray technique was used to detect HPV. The HPV 56 subtype was observed in the cervical smear and HPV 6 in dental biofilm. CONCLUSION: In this pregnant adolescent, HPV infection was present in both the cervix and the mouth, but the HPV subtypes infecting these two areas were different.

CONTEXTO: A adolescência e a gestação são consideradas fatores de risco para a infecção pelo papilomavírus humano (HPV). A relação entre essa infecção no colo do útero e na boca num mesmo paciente é controversa. RELATO DE CASO: Descrever o caso de uma adolescente grávida de 16 anos que apresentou a infecção pelo HPV no colo do útero e na boca. Esfregaços foram realizados no colo do útero e em língua/palato. Amostras de biofilme dental também foram coletadas. Para detectar o HPV, foi utilizada a técnica do microarranjo. O HPV 56 foi o subtipo encontrado no esfregaço cervical e o tipo HPV 6 no biofilme dental. CONCLUSÕES: Observamos, nessa adolescente grávida, a presença do HPV na boca e no colo do útero, mas os subtipos virais que infectavam essas duas regiões eram distintos.

Sobre-expresión de la proteina p16 en biopsias con diagnóstico de NIC I, positivas para genoma de papiloma virus humano. Instituto del Cáncer. Solca Cuenca-Ecuador / Over - expression of p 16 protein in biopsies with diagnosis of NIC I, positive for genome of human papilloma virus. Institute Of Cancer. Solca Cuenca - Ecuador. 2009 - 2010

Numerosos estudios mencionan que la sobre-expresión de la proteína p16, un marcador biológico que permite identificar lesiones pre-neoplásicas del epitelio exocervical, tendría una alta asociación con el Papiloma Virus Humano (HPV) de alto riesgo oncogénico. Es un estudio descriptivo correlacional cuyo objeti-vo fue establecer asociación de las Neoplasias Intraepiteliales Cervicales grado I (NIC I), HPV positivos, con la expresión del p16.Materiales, métodos y resultados: Es un estudio correlacional que se realizó en el período de noviembre de 2009 a noviembre de 2010; se presentaron 256 casos de NIC I de los cua-les, 72 fueron HPV positivos; se practicó técni-ca de p16. La edad promedio de las mujeres fue de 41 años. Se encontró positividad para el p16 en 40 casos (55.6%) y fueron negativos 32 (44.4%). De los casos positivos para p16, los tipos virales más frecuentes fueron los de alto riesgo: 33 (82.5%). El p16 fue valorado en cuantía, distribución e intensidad, estableciéndose relación entre la intensidad del p16 con los virus de alto riesgo (p=0.038). Cuan-do se analizó la edad y el tipo viral, pacientes entre 20 y 40 años (36 casos, 90%) presenta-ron genoma de HPV de alto riesgo.Conclusiones: Existió correlación entre la intensidad del p16 con la presencia de HPV de alto riesgo, ayudando a seleccionar grupos con tendencia a la progresión de la enfermedad.

Numerous studies report that overexpression of p16 protein that is a biomarker to identify premalignant exocervical epithelium lesions would have a high association with human papillomavirus (HPV) with high oncogenic risk. It is a descriptive correlational study whose objective was to establish association of Cer-vical Intraepithelial Neoplasia grade I (CIN I), HPV positive, with the p16 expression.Materials, methods and results: It is a corre-lational study conducted from November 2009 to November 2010; about 256 cases of CIN I were presented, 72 were positive HPV; also p16 technique was performed. The wo-men average age was 41 years. There were 40 (55.6%) positive cases for p16, and only 32 (44.4%) were negative. From the positive ca-ses for p16, the most common viral types were high-risk patients: 33 (82.5%). The p16 was va-lued in amount, distribution and intensity, es-tablishing a relationship between the intensity of p16 with high-risk virus (p = 0.038). When the age and viral type were analyzed, patients between 20 and 40 years (36 cases, 90%) pre-sented HPV genome with high risk.Conclusions: There is correlation between the intensity of p16 with the presence of HPV high risk, helping select groups prone to disease progression.

Unexpected inverse correlation between Native American ancestry and Asian American variants of HPV16 in admixed Colombian cervical cancer cases.

BACKGROUND: European (E) variants of HPV 16 are evenly distributed among world regions, meanwhile Non-European variants such as European-Asian (EAs), Asian American (AA) and African (Af) are mostly confined to Eastern Asia, The Americas and African regions respectively. Several studies have shown that genetic variation of HPV 16 is associated with the risk of cervical cancer, which also seems to be dependent on the population. This relationship between ethnicity and variants have led to the suggestion that there is co-evolution of variants with humankind. Our aim was to evaluate the relationship between the individual ancestry proportion and infection with HPV 16 variants in cervical cancer. METHODS: We examined the association between ancestry and HPV 16 variants in samples of 82 cervical cancer cases from different regions of Colombia. Individual ancestry proportions (European, African and Native American) were estimated by genotyping 106 ancestry informative markers. Variants were identified by PCR amplification of the E6 gene, followed by reverse line blot hybridization (RLB) with variants specific probes. RESULTS: Overall European (E) and Asian American (AA) variants frequency was 66.5% and 33.5% respectively. Similar distribution was observed in cases with higher proportions of European or African ancestry. A higher Native American ancestry was significantly associated with higher frequency of E variants (median ancestry>23.6%, Age and place of birth adjusted OR: 3.55, 95% CI: 1.26-10.03, p=0.01). Even further, an inverse geographic correlation between Native American ancestry and frequency of infections with AA variants was observed (ρ=-0.825, p=0.008). Regions with higher proportion of Native American ancestry had a lower frequency of AA variants of HPV 16. CONCLUSIONS: This study suggests replacement of AA variants by E variants of human papillomavirus 16 in cervical cancer cases with high Native American ancestry.

Prognostic value of DNA and mRNA e6/e7 of human papillomavirus in the evolution of cervical intraepithelial neoplasia grade 2.

OBJECTIVE: This study aimed at evaluating whether human papillomavirus (HPV) groups and E6/E7 mRNA of HPV 16, 18, 31, 33, and 45 are prognostic of cervical intraepithelial neoplasia (CIN) 2 outcome in women with a cervical smear showing a low-grade squamous intraepithelial lesion (LSIL). METHODS: This cohort study included women with biopsy-confirmed CIN 2 who were followed up for 12 months, with cervical smear and colposcopy performed every three months. RESULTS: Women with a negative or low-risk HPV status showed 100% CIN 2 regression. The CIN 2 regression rates at the 12-month follow-up were 69.4% for women with alpha-9 HPV versus 91.7% for other HPV species or HPV-negative status (P < 0.05). For women with HPV 16, the CIN 2 regression rate at the 12-month follow-up was 61.4% versus 89.5% for other HPV types or HPV-negative status (P < 0.05). The CIN 2 regression rate was 68.3% for women who tested positive for HPV E6/E7 mRNA versus 82.0% for the negative results, but this difference was not statistically significant. CONCLUSIONS: The expectant management for women with biopsy-confirmed CIN 2 and previous cytological tests showing LSIL exhibited a very high rate of spontaneous regression. HPV 16 is associated with a higher CIN 2 progression rate than other HPV infections. HPV E6/E7 mRNA is not a prognostic marker of the CIN 2 clinical outcome, although this analysis cannot be considered conclusive. Given the small sample size, this study could be considered a pilot for future larger studies on the role of predictive markers of CIN 2 evolution.

Experiencia vivida de mujeres al ser diagnosticadas con virus del papiloma humano / Lived experience of women being diagnosed with human papiloma virus

Resumen:

Introducción: desde la visión médica se considera que las mujeres detectadas con el virus del papiloma humano (VPH) no siguen el tratamiento médico porque no saben las consecuencias para su salud o porque no quieren hacerlo. Esta visión deja de lado el sentir y la visión de la paciente sobre su proceso salud-enfermedad.

Metodología: estudio cualitativo que utiliza un enfoque o una aproximación fenomenológica para describir la experiencia de mujeres diagnosticadas con VPH. Se entrevistó a profundidad a 20 mujeres diagnosticadas (saturación de la teoría). Las informantes clave se identificaron en el listado nominal de la Clínica de Lesiones Tempranas del Hospital General de Ometepec, Guerrero. Las que aceptaron participar firmaron consentimiento informado. Para el análisis de datos se utilizó el método de Giorgi.

Resultados: las experiencias vividas de las mujeres diagnosticadas con el VPH se ubican entre la muerte física y su faceta como procreadoras, ya que el significado social de su sexualidad se centra en su función de mujer-madre. La promiscuidad de su pareja la consideran una actitud normal. El no usar condón es una cuestión que se establece ante la necesidad de ser reproductivas. Conclusiones: la significación sociocultural que tiene la mujer como procreadora limita el uso del condón. Su definición social y cultural la ubica como alguien sin derecho a conocer su cuerpo, el goce de su sexualidad y el respeto por su cuerpo y su salud, a ser única responsable de la integración de la familia y el cuidado de los hijos.

Abstract:

Introduction: From the medical point of view, women diagnosed with human papilloma virus (HPV) are noncompliant with treatment either because they are ignorant of the consequences for their health or because they simply do not want to comply. This view ignores the feelings and point of view of the patients about their health-illness process.

Methodology: This qualitative study used a phenomenological focus approach to describe the experience of women diagnosed with HPV. In-depth interviews were conducted with 20 women diagnosed with this virus. The key informants were identified from the Clínica de Lesiones Tempranas (Early Lesions Clinic) list at Ometepec General Hospital, in Guerrero, and signed informed consents. Giorgi's method was used for data analysis.

Results: The lived experiences of the women diagnosed with HPV are focused on physical death, and as reproductive women the social significance of their sexuality is centered on their function as wife-mother. The promiscuity of their partner is considered normal. Not using a condom is due to the need of being reproductive.

Conclusions: The sociocultural significance that the women have as procreators limits their use of condoms. Their social and cultural definition limits their right to know their body, to enjoy their sexuality, and their health. They are responsible for the family and for childcare.

Conhecimento e atitude de usuários do SUS sobre o HPV e as vacinas disponíveis no Brasil / SUS users? knowledge of and attitude to HPV virus and vaccines available in Brazil / SUS users? knowledge of and attitude to HPV virus and vaccines available in Brazil

OBJETIVO : Analisar o conhecimento de homens e mulheres acerca do HPV e das vacinas e sua intenção de serem vacinados e de vacinarem seus filhos adolescentes. MÉTODOS : Estudo descritivo, de corte transversal, com 286 mulheres (18 a 49 anos) e 252 homens (18 a 60 anos), usuários de cinco unidades básicas de saúde e duas policlínicas do Sistema Único de Saúde, em Campinas, SP, em 2011. Foi realizada entrevista estruturada. Realizou-se análise bivariada e regressão de Poisson para identificar variáveis associadas ao conhecimento sobre HPV e vacinas e à intenção de vacinação. RESULTADOS : Quase 40,0% dos entrevistados referiram ter ouvido falar do HPV e 28,9% mencionaram informações adequadas; a principal fonte de informação foi a mídia (41,7%); 8,6% tinham ouvido falar das vacinas. Depois de informados da existência das vacinas, cerca de 94,0% dos participantes disseram que se vacinariam e/ou vacinariam filhos adolescentes se as vacinas estivessem disponíveis na rede pública de saúde. Escolaridade > 8 anos e ser do sexo feminino estiveram independentemente associados a ter ouvido falar do HPV e das vacinas e a ter conhecimento adequado sobre o vírus. Maior idade associou-se a ter ouvido falar das vacinas. Não houve variáveis associadas à intenção de vacinação. CONCLUSÕES : Os resultados reforçam a necessidade de haver intervenções educativas na população para prover informação adequada sobre o HPV e sobre medidas de prevenção. .

Objetivo : Analizar el conocimiento de hombres y mujeres sobre el VPH y de las vacunas y su intención de ser vacunados y de vacunar sus hijos adolescentes. Métodos : Estudio descriptivo, de corte transversal, con 286 mujeres (18 a 49 años) y 252 hombres (18 a 60 años), usuarios de cinco unidades básicas de salud y dos policlínicas del Sistema Único de Salud, en Campinas, SP – Brasil, en 2011. Se realizó entrevista estructurada. Se utilizó el análisis bivariado y regresión de Poisson para identificar variables asociadas al conocimiento sobre VPH y vacunas y la intención de vacunación. Resultados : Casi el 40,0% de los entrevistados mencionaron haber oído hablar del VPH y 28,9% relataron informaciones adecuadas; la principal fuente de información fueron los medios de comunicación (41,7%); 8,6% habían oído hablar de las vacunas. Después de informados de la existencia de las vacunas, cerca de 94,0% de los participantes dijeron que se vacunarían y/o vacunarían hijos adolescentes si las vacunas estaban disponibles en la red pública de salud. Escolaridad > 8 años y ser del sexo femenino estuvieron independientemente asociados al haber oído hablar del VPH y de las vacunas y tener conocimiento adecuado sobre el virus Mayor edad se asoció al haber oído hablar de vacunas. No hubo variables asociadas a la intención de vacunación. Conclusiones : Los resultados refuerzan la necesidad de haber intenciones educativas en la población para proveer información adecuada sobre el VPH y sobre medidas de prevención .

OBJECTIVE : To investigate knowledge of HPV and HPV vaccines in men and women, users of the Brazilian Unified Health System, and the intention to get themselves and their teenage children vaccinated. METHODS : A descriptive, cross-sectional study with 286 women (18-49 years old) and 252 men (18-60 years old), users of five primary health units and two polyclinics in Campinas, SP, Southeastern Brazil, was carried out. Participants were interviewed in 2011 using a structured questionnaire. Bivariate and Poisson regression analysis were performed to identify variables associated with knowledge of HPV and HPV vaccines, and participants vaccination intentions. RESULTS : Almost 40.0% of the participants reported having heard of HPV and 28.9% mentioned adequate information. The main information source was the media (41.7%). Only 8.6% of the participants had heard of the HPV vaccines. Once the participants were informed of the existence of HPV vaccines about 94% of them said they would get vaccinated and/or vaccinate their teenage children, if the vaccines were available in the public health system. Schooling of over 8 years and being female were the variables independently associated with having heard of HPV, the vaccines and having adequate knowledge of the virus. Advanced age was associated with having heard of HPV vaccines. There were no variables associated with the vaccination intentions. CONCLUSIONS : These results reinforce the need for educational activities that provide the population with adequate information on HPV and preventive measures. .