HPV genotyping and p16 expression in Xingu Indigenous Park, Brazil.

The association between high-risk human papillomavirus (HPV) genotypes and p16 expression in indigenous women from the Xingu Indigenous Park, Brazil, was unknown. This study evaluated p16 expression in women with a histological diagnosis of cervical intraepithelial neoplasia (CIN) 3 or higher and correlated this expression with HPV genotypes to determine possible discrepancies in the expression of this marker. We evaluated 37 previously collected samples with different HPV genotypes and high-grade lesions diagnosed based on cytology, histology, and colposcopy. Immunohistochemical analysis was performed using paraffin-embedded tissue sections and the CINtec® Histology Kit. p16 protein expression was investigated by immunostaining with an anti-p16 antibody. HPV genotyping was performed by reverse hybridization. The age of the study population ranged from 22-75 years (43.81 ± 15.89 years) and parity ranged from 1-11 (5.92 ± 2.58). Thirteen different HPV genotypes were found using the INNO-LiPA kit. Single and multiple infections by HPV were found with prevalence of single infections (P = 0.029). Comparison between HPV genotype and simple or multiple infections was highly significant; it was observed more HPV 52 followed by HPV 16 in single infections (P < 0.001). p16 expression was predominantly diffuse, which was observed in 91.7% of lesions, whereas 8.3% were focal (P < 0.001). HPV 52, HPV 16 and 31 were the most prevalent HPV types in high-grade CIN in these indigenous women. Diffuse p16 expression in high-grade CIN was not influenced by the viral genotype; however, more studies are necessary to further our understanding of this restricted group.

HPV infection-associated anogenital cyto-colpo-histological findings and molecular typing in HIV-positive women.

HIV and human papillomavirus (HPV) coinfection is increasing, especially in the anal canal (AC) and cervico-vaginal regions. We identified anal epithelium abnormalities related to high-risk HPV (HR-HPV) lesions in the lower genital tracts (LGTs) of HIV-positive women, described the HPV genotypes identified, and assessed the expression of E6/E7 oncogenes in coinfected patients. Ninety-eight women were enrolled in groups combining HIV status and presence or absence of HPV in the LGT. Anal and cervical smears were collected for cytology and HR-HPV assays using Cobas(®) and/or PapilloCheck(®). Samples with highly oncogenic HPV genotypes were confirmed by NucliSENS EasyQ(®). Forty-two HIV-positive (25-52; mean age 39.5) and 56 HIV-negative (18-58; mean age 35.7) patients were included. E2 and C1 groups presented AC alterations (P = 0.002); altered images for high-resolution anoscopy were higher in E1 and C2 (P < 0.001). Of the 29 women with alterations, 41.38% were HIV-negative and 58.62% were HIV-positive (P < 0.001). HIV-positive patients accounted for 29% of the anal high-grade squamous intraepithelial lesions (P = 0.015). The Cobas(®) positive result frequency was higher in three AC groups than in the other groups. There was variation in the number of HPV types in the cervico-vaginal samples among the study groups (P < 0.001). Anal cytology and anoscopy showed more altered findings in HIV-positive patients with HPV in the LGT. HR-HPV anal infections by various genotypes are common and are associated with cervical infections in HIV-positive patients. E6/E7 expression is apparently more common in the AC of HIV-positive women.

Accuracy of the cytopathology, bacterioscopy, and vaginal flora culture.

INTRODUCTION: An over-population of vaginal microorganisms causing inflammatory processes renders it difficult to properly assess the cytopathological exam that aims to screen precedent cervical lesions. On the contrary, the occurrence of the microbial flora saprophyte does not influence correct cythodiagnosis. OBJECTIVE: To assess the composition of vaginal tract aerobic microorganisms of asymptomatic women in menacme and post-menopause, and to analyze the accuracy of cytopathologic, bacterioscopic exams, and culturing of the flora. METHODS: The women were first submitted to a focused anamnestic interrogatory and then submitted to gynecological exam. A sample of the vaginal fluid was collected with a culture swab and a smear was made on two glass slides for stained bacterioscopic exam (GRAM). The collection of material was then compiled in a cytopathologic smear analysis. All women signed the free and informed consent letter and the project was approved by the Ethics Research Board of Hospital São Paulo - UNIFESP. RESULTS: Bacterioscopy and culture proved to be better than the cytopathologic exam in featuring the bacilli and cocci. The bacterioscopy provided a better detection of the presence of bacilli (p < 0.001); no statistical difference was seen between both exams with respect to the detected cocci. The beta-hemolytic Streptococcus group was of significance in post-menopausal women (p < 0.05). CONCLUSION: In this study, the bacterioscopic and culture exams of the vaginal fluid were more effective in assessing the vaginal flora and in the detection of bacilli, compared to the cytopathological exam.

Depth of glandular crypts and its involvement in squamous intraepithelial cervical neoplasia submitted to large loop excision of transformation zone (LLETZ).

BACKGROUND: The authors aimed to confirm the depth of six mm in order to achieve an optimal eradication of the lesion. MATERIALS AND METHODS: This is a retrospective observational study of 94 cervical surgical pieces from women aged 17 to 22 years with a cyto-colpo-histopathological diagnosis of high-grade squamous cervical intraepithelial neoplasia (CIN II and/or CIN III) submitted to large loop excision of transformation zone (LLETZ). The glandular crypts and margins, both exposed or not to CIN, were assessed. The compromise and the maximum depth of the glandular crypts were noticed. RESULTS: After LLETZ, 23 (24.47%) cases presented a neoplasic impairment of endocervical margin and ten (10.64%) of the ectocervical margin. The largest noticed crypt measured 4.500 mm and the shortest 0.100 mm, with an average of 2.148 mm. CONCLUSIONS: Squamous CIN more frequently show the exposure of surgical margins to LLETZ. The deeper location of glandular crypts in the cases studied was 4.500 mm, while the largest neoplastic extension was 3.000 mm.The therapeutic method depends on this knowledge.

Immunohistochemical expression of granzyme B and vascular endothelial growth factor (VEGF) in normal uterine cervices and low and high grade squamous intraepithelial lesions.

PURPOSE: This study aimed to evaluate the immunoexpression of granzyme B and vascular endothelial growth factor (VEGF) in the variants of cervical squamous intraepithelial neoplasia. METHODS: Granzyme B immunohistochemical expression was studied in the epithelium, stroma and in both the epithelium + stroma of 142 fragments of uterine cervix; there were 34 grade 1 cervical intraepithelial neoplasias (CIN 1), 36 grade 2 cervical intraepithelial neoplasias (CIN 2), 33 grade 3 cervical intraepithelial neoplasias (CIN 3) and 39 uterine cervix fragments without abnormalities - control group. Immunoexpression of VEGF was studied in 160 uterine cervix fragments, with 43 grade 1 cervical intraepithelial neoplasias (CIN 1), 33 grade 2 cervical intraepithelial neoplasias (CIN 2), 31 grade 3 cervical intraepithelial neoplasias (CIN 3) and 53 uterine cervix fragments without abnormalities--control group. RESULTS: In the stroma, immunoexpression of granzyme B in grade 1 cervical intraepithelial neoplasias was smaller than in grade 3 cervical intraepithelial neoplasias. High VEGF immunoexpression was found in grade 3 cervical intraepithelial neoplasias while it was low in grade 1 cervical intraepithelial neoplasias and in the control group. CONCLUSION: The higher the severity of the cervical intraepithelial lesion, the higher the immunoexpression of granzyme B. A progressive increase in VEGF immunoexpression was found in the intense grade, according to the severity of the cervical intraepithelial neoplasia.

Low-dose 5-fluorouracil adjuvant in laser therapy for HPV lesions in immunosuppressed patients and cases of difficult control.

The authors established a protocol for the use of 5-fluorouracil (5FU) adjuvant in lasertherapy for clinical and subclinical HPV infection in immunosuppressed patients, persistent lesions and as reinforcement treatment in cases of poor progress. Sixty-four patients were evaluated, of whom 26 were immunosuppressed, 34 presented persistent lesions and four received intravaginal reinforcement treatment with 2.5 g 5% 5FU every two weeks, or biweekly vulvar reinforcement after lasertherapy. On average, five 5FU courses were used, but in the immunossuppressed patients its use was maintained indefinitely. The rate of complete response was 66%, but the immunossuppressed patients showed less response (46.2%) when compared with the persistent lesion/reinforcement treatment group (78.9%). The responses were positive in the two groups when compared to that with no response. We deem the use of low-dose 5FU an excellent alternative in cases of difficult HPV progress, presenting a low cost and minimal side-effects.