Renata Mirian Nunes Eleutério

Associação entre a carga viral de HPV de alto risco, expressão de p16INK4a e lesões intra-epiteliais escamosas do colo uterino

OBJECTIVE: To determine the (HR-HPV) high risk HPV viral load in squamous intra-epithelial lesions and association with p16INK4a expression. METHODS: A series of 109 cervical biopsies were studied (57 normal tissue, 26 low grade squamous intra-epithelial lesions [LSIL] and 26 high grade squamous intra-epithelial lesions [HSIL]). Detection of high risk HPV and viral load in cervical cells was made by molecular biology using hybrid capture 2nd generation collected before the biopsy. The p16INK4a was identified by immunohistochemistry using the p16INK4a kit (clone E6H4). RESULTS: High risk HPV was positive in 57.8% of all cases (29.8% in normal tissue, 80.8% in LSIL and 96.1% in HSIL). Protein p16INK4a was expressed in 23.8 % of squamous intra-epithelial lesions (15.4% in LSIL and 84.6% in HSIL). In normal tissue all cases were negative to p16INK4a. The viral load was higher in p16 positive cases than in negative cases (positive p16INK4a mean of 669.9 RLU/PCB [9.47 - 2814.9] and negative p16INK4a mean of 253.94 RLU/PCB [1.07 1882.21] (p<0.05). However when studying just the HSIL cases differences were not significant. CONCLUSION: In this study although the HR-HPV viral load had shown a significant difference between p16 positive and negative cases, in HSIL cases this finding was not confirmed. New studies with a larger number of cases are necessary for consistent conclusions.

Identificação de DNA-HPV em adolescentes e mulheres jovens sem coito vaginal

Introduction: the human papillomavirus (HPV) virus is an extremely prevalent in the world. Currently there are around 200 types identified, about 45 of which affect the anogenital area. Although the high prevalence of HPV infection among sexually active adolescents is known, few studies have succeeded in demonstrating the presence of the virus among girls before sexual intercourse. Objective: to assess the prevalence of human papillomavirus (HPV) in adolescents and young women prior to coitarche assisted at an outpatient gynecological unit. Methods: this study included 50 adolescents assisted at the gynecology unit in University Hospital Pedro Ernesto (HUPE), State University of Rio de Janeiro. The patients were below 20 years old, who reported no sexual intercourse and with intact hymen on physical examination by a gynecologist with broad experience, from January 2007 to January 2009. The material for study was collected from vulvar vestibule for detection of HPV-DNA by Hybrid Capture second generation (hC2). Results: in the 50 patients tested for HPV-DNA age ranged from 11 to 20 years old (mean: 15.88 + 2.04) and all of them denied coitus and had intact hymen. The hC2 was positive in three cases (6%). In two cases non-high risk HPV was detected and in one case high-risk HPV was identified. After 6 months, a second test for HPV-DNA was negative in two out of three patients. The third one was not tested. Conclusion: HPV infection can occur before the first sexual intercourse and even in situations where there is no genital contact.

Liquid‐based cytology and HPV DNA testing using intra‐anal specimens from HIV‐negative women with and without genital HPV‐induced lesions

Screening for anal cancer using cytology has not been considered in immunocompetent women. The aim of this study was to identify cytological atypia and human papillomavirus (HPV) DNA in intra‐anal specimens from human immunodeficiency virus (HIV)‐negative women with and without genital HPV lesions.

The risk of high-grade squamous intraepithelial lesions in women with low serum levels of vitamin A.

Background: The progression of the cervical squamous intraepithelial lesion (SIL) is associated with many cofactors, and nutritional aspects are gradually assuming an important role in understanding the physiopathogenesis. Objective: To assess the serum levels of retinol in women from a poor region of Brazil with a histological diagnosis of cervical high-grade SIL (HSIL). Methods: A cross-sectional study was conducted in 62 women aged 14-48 years who had a cervical biopsy and were treated at the Federal University of Ceara in Fortaleza, Brazil. Serum retinol levels were measured in peripheral blood by a spectrophotometry technique. Values were categorized as subnormal (<20 µg/dl) or normal (≥20 µg/dl). Multivariate logistic regression was used to determine the association between serum levels of retinol and HSIL for a confidence interval of 95%. Results: Twenty-six out of 62 women (42%) had a negative histopathological result for SIL and 36 (58%) were diagnosed with SIL (20 low SIL and 16 high SIL). The level of retinol was more frequently low in women with HSIL, but there was no statistical significance [p = 0.409, OR: 2.26 (0.33-15.59)]. Conclusion: There was no association between high SIL and low levels of retinol in peripheral blood.

HPV-DNA Hybrid Capture Test

OBJECTIVE To evaluate if the cellularity of Hybrid Capture samples (Digene, Sao Paulo, Brazil) influences the results of HPV-DNA Hybrid Capture tests in men. STUDY DESIGN We harvested material from penile scrapings for the Hybrid Capture HPV test. This material was then used to make cytologic smears, which we used to evaluate for the presence of nonnucleated squamous cells, nucleated squamous cells and glandular cells. The cellularity of nucleated squamous cells was classified as absent, low, moderate or high. Subsequently, we performed the Hybrid Capture test to identify the low and high risk of HPV and compared these results with the cytologic findings. We used the Fisher and odds ratio tests at CI of 95% to determine statistical significance. RESULTS Of the 88 tests performed, 65 (74.0%) were negative for HPV-DNA and 23 (26.0%) were positive. Nucleated and nonnucleated squamous cells were absent on nine slides, all of which tested negative for HPV. When only nonnudcleated squamous cells were found, 20% of the cases were positive for HPV-DNA (p < 0.0001; OR = 26.185). The presence of nucleated squamous cells correlated with 33% HPV-DNA positivity (p < 0.0001, OR = 49.05). CONCLUSION Assessing the presence of non-nucleated and nucleated squamous cells on cytologic smears prior to performing an HPV-DNA test is a useful tool for quality control in penile samples.

Atypical Squamous Cells: Cytopathological Findings and Correlation with HPV Genotype and Histopathology

Objective: We aimed to assess potential associations between atypical squamous cell (ASC) subgroups: ASC-US (undetermined significance) and ASC-H (cannot exclude high-grade squamous intraepithelial lesion), regarding cytomorphological features, high-risk (HR) human papillomavirus (HPV) testing, and histological outcomes in a sample of Brazilian women. Study Design: Cross-sectional study which evaluated 1,346 liquid-based cytologies between January 2010 and July 2016 with ASC results. ASC-US and ASC-H were analyzed for frequency, diagnostic criteria, and cytological findings and compared with HR-HPV tests and histological outcomes. Results: Enlarged nucleus was the most frequent ASC-US criterion, but alternative criteria were present in 20% of the total cases. No ASC-US criteria were associated with histological outcomes or HR-HPV positivity. Parakeratosis, corneal pearl, giant cells, and binucleation were strongly associated with ASC-US while hyperkeratosis was associated with high-grade squamous intraepithelial lesions (HSIL) or a superior outcome. HR-HPV was positive in 64.39% of ASC-US and 65.38% of ASC-H. HSIL or superior outcomes also occurred in 13.33% of ASC-US and 64.71% of ASC-H cases. Conclusion: Alternative criteria for ASC-US were relatively frequent. Reactive cellular changes suggestive of atypias were more abundant in ASC-US. Although ASC-H is associated with worse histological outcomes, no differences in HPV positivity were found in comparison to ASC-US.

Atypical Squamous Cells in Liquid-Based Cervical Cytology: Microbiology, Inflammatory Infiltrate, and Human Papillomavirus-DNA Testing

Objective: The aim of this study was to assess the correlation between atypical squamous cells (ASC) and inflammatory infiltrate and vaginal microbiota using cervical liquid-based cytological (SurePath®) and high-risk human papillomavirus (HR-HPV) tests. Study Design: A cross-sectional study was conducted using a 6-year database from a laboratory in Fortaleza (Brazil). Files from 1,346 ASC cases were divided into subgroups and results concerning inflammation and vaginal microorganisms diagnosed by cytology were compared with HR-HPV test results. Results: An absence of specific microorganisms (ASM) was the most frequent finding (ASC of undetermined significance, ASC-US = 74%; ASC - cannot exclude high-grade squamous intraepithelial lesion, ASC-H = 68%), followed by bacterial vaginosis (ASC-US = 20%; ASC- H = 25%) and Candida spp. (ASC-US = 6%; ASC-H = 5%). Leukocyte infiltrate was present in 71% of ASC-US and 85% of ASC-H (p = 0.0040), and in these specific cases HR-HPV tests were positive for 65 and 64%, respectively. A positive HR-HPV test was relatively more frequent when a specific microorganism was present, and Candida spp. was associated with HR-HPV-positive results (p = 0.0156), while an ASM was associated with negative HR-HPV results (p = 0.0370). Conclusion: ASC-US is associated with an absence of inflammation or vaginosis, while ASC-H smears are associated with Trichomonas vaginalis and inflammatory infiltrate. A positive HR-HPV is associated with Candida spp. in ASC cytology.

Inflammatory cells in liquid‐based cytology smears classified as bacterial vaginosis

Bacterial vaginosis (BV) remains an enigmatic question. The term “osis” instead of “itis” is used because commonly, there are no inflammatory process associated with BV. However, leukocytes are often observed in it.

Prevalence of HPV in Adolescents Virgins and Sexually Active at a University Hospital in the City of Rio de Janeiro, Brazil

Sexually transmitted diseases (STDs) are common worldwide, with especially alarming numbers in Brazil. Among the most common infections is human papillomavirus (HPV). The possibility of the nonsexual transmission of HPV is not well defined and is the subject of debate. This study aimed to identify the prevalence of HPV in adolescents with no history of sexual intercourse compared with a group of similar age with sexual activity. 100 adolescents were evaluated with at least two years after menarche, who attended from January 2007 to January 2009 at the University Hospital Pedro Ernesto, Rio de Janeiro, Brazil. Among the adolescents, 50 had intact hymen and 50 reported regular sexual activity. For patients without sexual intercourse (group 1) we collected material from vestibule and for patients with sexual activity (group 2) we collected material from vagina and endocervix. The search for HPV-DNA 2nd generation hybrid capture (hC2) was performed. In group 1 the test was positive in 3 cases (6%). In the second group, 33 cases (66%) were positive for at least one site. The positivity in girls with sexual activity is high. On the other hand, the HPV infection, although rare, may occur in girls without sexual intercourse.