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Background The objective of this study was to estimate the prevalence of Human papillomavirus (HPV) genotypes and Chlamydia trachomatis (CT) infections among adolescents and young adult women and to identify the risk factors associated. Methods This study included 276 sexually active participants, classified as adolescents (15-19 years) and young adult women (20-24 years) that realized conventional cytology and were tested for 27 HPV genotypes and for CT. Bivariate and multivariate analyses were performed to evaluate the factors associated with both infections. Results The prevalence of HPV infection was 50.7% in adolescents and 43.0% in young adult women. The HPV-16 was the most prevalent (27.7%), followed by HPV-68 (6.9%), HPV-52 (6.2%), HPV-39 (4.6%), and HPV-73 (4.6%). The prevalence of CT infection was 11.5% among adolescents and 6.2% among young adult women. Cytological abnormalities were found in 14.2% among adolescents, of which 80.9% were positive for HPV and 10% of young adult women of which 84.6% were positive for HPV infection. HPV16 and HPV 68 were detected in 35.7% and 32.1% of cases with cytological abnormalities. Factors independently associated with HPV infection obtained by multivariate analysis were unmarried or divorced marital status and the presence of cytological abnormalities. The same variables were analyzed for the association with CT infection, and the association with cytological abnormalities remained significant. Conclusion HPV and CT infections are very prevalent in adolescent and young adult women; these findings reinforce the need of early vaccination, prior to the onset of sexual activity, and justify implementation of molecular screening tests.

Because of its role in preserving DNA/RNA stability, there is an increasing number of studies addressing the relationship between Fused in Sarcoma (FUS) expression and cancer. However, the disparity in FUS/FUS oncogenic vs. tumor-suppressive roles may be attributed to the complex molecular pathways associated with FUS regulation in different cancer types, and its role in cervical carcinogenesis remains largely unexplored. Methods: We determined FUS protein expression in specimens of 61 patients with advanced cervical cancer. Long-term (> 10 years) clinical follow-up data for these patients were available, and we determined disease-free, cancer-related and overall survival as related to FUS expression. Results: There were no significant associations between FUS expression and patients’ age, tumor grade, and acute/late toxicity events related to treatment (either radiation alone or chemoradiation). However, multivariate Cox regression analysis for disease-free survival (recurrence), overall survival (death) and cancer-related survival showed that patients with high average FUS expression fared significantly better than their counterparts with low average FUS expression, both in terms of disease-free survival (HR = 0.31; 95%CI 0.12 to 0.77; p = 0.01) and cancer-related survival (HR = 0.41; 95%CI 0.17 to 0.98; p = 0.04). Conclusions: Our study shows that high FUS protein expression in advanced cervical cancer specimens is a potent harbinger of better prognosis, and can as such be used in clinical practice to help characterize patients and, possibly, plan treatment and follow-up strategies.

Background Expression of matrix metalloproteases 2, 9 and 14 (MMP-2, MMP-9, MMP-14), tissue inhibitors of metalloprotease 1 and 2 (TIMP-1, TIMP-2) and vascular endothelial growth factor A (VEGF-A) is involved in tumor invasion and metastasis via extracellular matrix degradation and angiogenesis. This study aimed to assess whether the expression of MMP-2, MMP-9, MMP-14, TIMP-1, and TIMP-2 in tumors and in the adjacent stroma is associated with cervical cancer prognosis. Methods This study analyzed a retrospective cohort of 64 patients. Protein expression was previously obtained by immunohistochemistry from biopsies containing both tumor and stroma. The expression and percentage of stained cells were categorized as high or low according to the cutoff points by using ROC curves. The follow-up data was collected from diagnosis to the last clinical visit. Clinical status categorized as alive without disease, alive with disease, death due to other causes, and death from the disease. The relative risk of death from the disease was evaluated according to the proteins expression using a cause-specific Cox regression model with a 95% confidence interval (95%CI). For the significant associations (p Results High expression levels of stromal MMP-2 (RR; 95%CI: 3.91; 1.17–13.02) and stromal TIMP-2 (RR, 95%CI: 8.67; 1.15–65.27) were associated with a greater relative risk of death from the disease and with lower survival (p = 0.03; p = 0.04) than lower expression levels. Low expression levels of stromal MMP-9 (RR, 95%CI: 0.19; 0.05–0.65) and tumoral MMP-9 (HR, 95%CI: 0.19; 0.04–0.90) were protective factors against death from the disease and were associated with poorer survival. Conclusions High expression levels of MMP-2 and TIMP-2 in the stroma were significantly associated with poor survival in cervical cancer patients. High expression of MMP-9 was associated with a favorable cervical cancer prognosis.

IntroductionHuman papillomavirus (HPV) infection is associated with the development of anogenital and head and neck cancers. In recent years a potential role of HPV in colorectal cancer (CRC) has been suggested.ObjectiveTo investigate the presence of HPV in colorectal carcinomas and to study the role of p16INK4a as a marker of transcriptionally active HPV infection. In addition, to investigate the correlation between these findings and the CRC prognostic factors.MethodsCase control study with 92 cases of colorectal cancers, 75 controls of normal tissue adjacent to the tumor, and 30 controls of precursor lesions, including polyps and colorectal adenomas. Paraffinized samples were used, HPV detection and genotyping were performed by PCR and reverse hybridization by using the INNO LIPA kit, with SPF10 plus primers. The expression of the p16INK4a protein was investigated using immunohistochemistry. Data analysis was performed using descriptive, univariate statistics and survival curves were calculated by using the Kaplan Meier and log-rank method.ResultsHPV was detected in 13% of the cases and the most prevalent genotype was HPV 16. HPV DNA was not detected in either control groups. The high expression of p16INK4a was observed in 30% of the cases, but it was not associated to the presence of HPV. The overall survival was 53.3% and was influenced by prognostic factors such as later stage, lymph node and distant metastasis.ConclusionsBased on these results, HPV is unlikely to be involved in colorectal carcinogenesis and p16INK4a expression is not a relevant marker of transcriptionally active HPV infection in CRC.

It is well known that persistent infection with high-risk HPV (hr-HPV), mostly HPV-16 and 18, is the main cause of cervical cancer development. Manganese superoxide dismutase (MnSOD or SOD2) are highly expressed in different neoplasia. The present study investigated SOD2 protein expression and the presence of hr-HPV types in 297 cervical samples including non-neoplastic tissue, cervical intraepithelial neoplasia grade 3 (CIN3), squamous cell carcinoma (SCC) and adenocarcinoma (ADC). Strong SOD2 expression was significantly higher in ADC (82%) than CIN3 (52%) or SCC (64%). There was no association between SOD2 expression and HPV 16 and/or 18 detection for every lesion analyzed. Binary Logist Regression revealed that strong SOD2 expression (OR: 27.50, 6.16-122.81) and HPV 16 and/or HPV 18 (OR: 12.67, 4.04-39.74) were independently more associated with CIN3 than non-neoplastic cervix. Strong SOD2 expression (OR: 3.30, 1.23-8.86) and HPV 16 and/or HPV 18 (OR: 3.51, 1.03-11.87) were independently more associated with ADC than SCC. Similar findings for SOD2 expression were observed by the Cochran Mantel-Haenszel test, controlling for HPV-16 and/or HPV 18. In conclusion, the expression of SOD2 was increased in CIN3 and SCC, and more increased in cervical ADC than in SCC. Strong SOD2 expression was statistically independent of the presence of HPV 16 and/or 18. These findings suggest that the mitochondrial antioxidant system and HPV infection could follow independent pathways in the carcinogenesis of cervical epithelium and in the differentiation to SCC or ADC of the cervix., Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP; Grants No. 2008/57889-1, 2010/07739-3 and 2012/16059-1) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)/Instituto Nacional de Ciência e Tecnologia das Doenças Associadas ao HPV (INCT-HPV) (CNPq Grant No. 573799/2008-3), info:eu-repo/semantics/publishedVersion

Background Studies have indicated that bacterial vaginosis (BV) might be a cofactor for the acquisition and persistence of high-risk papillomavirus, enabling the development of cytological abnormalities. The presence of endocervical and metaplastic cells makes the smear more adequate for the detection of these abnormalities once these cell types are representative of the transformation zone, a site of increased susceptibility to viral infection. Methods The purpose of this study was to evaluate the patterns of vaginal microbiota, the representation of endocervical and/or metaplastic cells, and the detection of cytological abnormalities in cervical smears from women 15 to 64 years old. Results from satisfactory cytological smears performed in a laboratory school from the Federal University of Goias were analyzed. The degree of association between the categorical variables was evaluated by the χ2 test, Fisher's Exact test, and stratified analysis through the estimation of the prevalence ratio, with 95% confidence intervals and 5% statistical significance level (P Results The global prevalence of BV and cytological abnormalities was 22.02% and 8.21%, respectively. BV and the representation of endocervical and/or metaplastic cells were independently associated with the detection of high-grade cytological abnormalities in the cervical smears of women between 25 and 64 years old. Conclusions BV and representation of endocervical and/or metaplastic cells were independently associated with the detection of high-grade cytological abnormalities reinforcing the importance of specimen adequacy and microbiota in the cervical microenvironment.

Alterations in specific DNA damage repair mechanisms in the presence of human papillomavirus (HPV) infection have been described in different experimental models. However, the global effect of HPV on the expression of genes involved in these pathways has not been analyzed in detail. In the present study, we compared the expression profile of 135 genes involved in DNA damage repair among primary human keratinocytes (PHK), HPV-positive (SiHa and HeLa) and HPV-negative (C33A) cervical cancer derived cell lines. We identified 9 genes which expression pattern distinguishes HPV-positive tumor cell lines from C33A. Moreover, we observed that Three Prime Repair Exonuclease 1 (TREX1) expression is upregulated exclusively in HPV-transformed cell lines and PHK expressing HPV16 E6 and E7 oncogenes. We demonstrated that TREX1 silencing greatly affects tumor cells clonogenic and anchorage independent growth potential. We showed that this effect is associated with p53 upregulation, accumulation of subG1 cells, and requires the expression of E7 from high-risk HPV types. Finally, we observed an increase in TREX1 levels in precancerous lesions, squamous carcinomas and adenocarcinomas clinical samples. Altogether, our results indicate that TREX1 upregulation is important for cervical tumor cells growth and may contribute with tumor establishment and progression., This research was supported by grants of FAPESP (2010/20002-0; 2008/57889-1) and CNPq (480552/2011-8); and fellowships for BP (CNPq 573799/2008-3; CAPES 1524553), WSA (FAPESP 2012/16512-8), MM (FAPESP 2011/14416-9), SH (2013/27006-9).

Background. Approximately 90% of all anal cancers are associated with human papillomavirus (HPV), especially high-risk genotypes such as HPVs 16 and 18. Objective. To investigate the clinical and prognostic aspects of anal cancers associated with the presence, as well as the genotypic distribution of human papillomavirus (HPV). Methods. A retrospective study carried out over a 10-year period, using clinical and molecular data, with PCR analysis and reverse hybridization (INNO-LIPA kit), in anal cancers. The data analysis was done using descriptive univariate statistics, and the survival curves were made using the Kaplan–Meier and log-rank methods. Results. Of the 81 formalin-fixed and paraffin-embedded specimens, HPV prevalence was 69% and was significantly higher in squamous cell carcinomas (SCC) than in other anal tumors (p=0.0001). Female patients had a higher prevalence of HPV (p=0.01). Multiple infections were detected in 14.3% of cases. The most prevalent genotypes were HPVs 16, 33, and 18. The overall survival at 60 months was 44.3%, and the prognostic factors included gender (p=0.008) with greater survival for men (52.9%) in comparison to women (29.6%), histological type (p=0.01), SCC (54.4%), adenocarcinomas (37.5%), other carcinomas (14.2%), and the presence of distant metastasis (p=0.01). Survival was not influenced by the presence of HPV (p=0.54). Conclusions. The association of HPV to anal cancer was found in this study, especially in SCC. However, the presence of HPV did not influence the prognosis of patients with anal cancer.

Background Penile carcinoma (PC) is a rare, highly mutilating disease, common in developing countries. The evolution of penile cancer includes at least two independent carcinogenic pathways, related or unrelated to HPV infection. Objectives To estimate the prevalence, identify HPV genotypes, and correlate with clinicopathological data on penile cancer. Methods A retrospective cohort study involving 183 patients with PC undergoing treatment in a referral hospital in Goiânia, Goias, in Midwestern Brazil, from 2003 to 2015. Samples containing paraffin embedded tumor fragments were subjected to detection and genotyping by INNO-LiPA HPV. The clinicopathological variables were subjected to analysis with respect to HPV positivity and used prevalence ratio (PR), adjusted prevalence ratio (PRa) and 95% confidence interval (CI) as statistical measures. Results The prevalence of HPV DNA in PC was 30.6% (95% CI: 24.4 to 37.6), high-risk HPV 24.9% (95% CI: 18.9 to 31.3), and 62.5% were HPV 16. There was a statistical association between the endpoints HPV infection and HPV high risk, and the variable tumor grade II-III (p = 0.025) (p = 0.040), respectively. There was no statistical difference in disease specific survival at 10 years between the HPV positive and negative patients (p = 0.143), and high and low risk HPV (p = 0.325). Conclusions The prevalence of HPV infection was 30.6%, and 80.3% of the genotypes were identified as preventable by anti-HPV quadrivalent or nonavalent vaccine. HPV infections and high-risk HPV were not associated with penile carcinoma prognosis in this study.

Vaginal infections may affect susceptibility to and clearance of human papillomavirus (HPV) infection and chronic inflammation has been linked to carcinogenesis. This study aimed to evaluate the association between bacterial vaginosis (BV) and inflammatory response (IR) with the severity of cervical neoplasia in HPV-infected women. HPV DNA was amplified using PGMY09/11 primers and genotyping was performed using a reverse line blot hybridization assay in 211 cervical samples from women submitted to excision of the transformation zone. The bacterial flora was assessed in Papanicolaou stained smears, and positivity for BV was defined as ≥ 20% of clue cells. Present inflammatory response was defined as ≥ 30 neutrophils per field at 1000× magnification. Age higher than 29 years (OR:1.91 95% CI 1.06-3.45), infections by the types 16 and/or 18 (OR:1.92 95% CI 1.06-3.47), single or multiple infections associated with types 16 and/or 18 (OR: 1.92 CI 95% 1.06-3.47), BV (OR: 3.54 95% CI 1.62-7.73) and IR (OR: 6.33 95% CI 3.06-13.07) were associated with severity of cervical neoplasia (CIN 2 or worse diagnoses), while not smoking showed a protective effect (OR: 0.51 95% CI 0.26-0.98). After controlling for confounding factors, BV(OR: 3.90 95% CI 1.64-9.29) and IR (OR: 6.43 95% CI 2.92-14.15) maintained their association with the severity of cervical neoplasia. Bacterial vaginosis and inflammatory response were independently associated with severity of cervical neoplasia in HPV-positive women, which seems to suggest that the microenvironment would relate to the natural history of cervical neoplasia.

Background Human papillomavirus (HPV) infection, bacterial vaginosis (BV), and cervicitis may play a role in the presence of cytological abnormalities in female adolescents and young women. This study aimed to estimate the prevalence of HPV, BV and cervicitis in female adolescents and young women and evaluate whether these conditions are associated with a finding of cytological abnormalities in cervical smears. Methods Cervical smears were screened using the conventional method and HPV-DNA detection was performed by PGMY-PCR. BV was defined as ≥20% clue cells in cervical smears. Cervicitis was assessed by counting leukocytes in five non-adjacent microscopic fields at 1000× magnification. Odds ratios and 95% confidence intervals were calculated. Results The study analyzed 251 samples, with cytological abnormalities being found in 9.5% (24/251). Atypical squamous cells of undetermined significance (50.0%) and low-grade squamous intraepithelial lesions (29.1%) were the most common findings. HPV, BV and cervicitis were found in 44.2% (111/251), 41.0% (103/251) and 83.2% (209/251) of cases, respectively. Of the variables investigated, BV and an abnormal cytological diagnosis were independently associated with HPV positivity. Cytological abnormalities were significantly associated with a finding of HPV and BV in the same woman, and also with a simultaneous finding of HPV, BV and cervicitis; however, the independent association of these combined variables did not remain significant in the multivariate analysis. Conclusions BV and cytological abnormalities were independently associated with HPV infection in female adolescents and young women. Diagn. Cytopathol. 2015;43:780–785. © 2015 Wiley Periodicals, Inc.

OBJETIVOS: Estimar a prevalência da vaginose bacteriana (VB), candidíase e tricomoníase e comparar os achados do exame físico da secreção vaginal com o diagnóstico microbiológico, obtido pelo estudo citológico do esfregaço vaginal, pelo do método de Papanicolaou. MÉTODOS: Estudo de corte transversal que incluiu 302 mulheres com idade entre 20 a 87 anos, submetidas à entrevista e exame ginecológico para avaliação da secreção vaginal e coleta de esfregaço citológico, no período de junho de 2012 a maio de 2013. Para avaliar a acurácia das características da secreção vaginal em relação ao diagnóstico microbiológico do esfregaço citológico foi empregado as análises de sensibilidade, especificidade, valor preditivo positivo (VPP) e valor preditivo negativo (VPN), com seus respectivos IC95%. Para avaliar o grau de concordância entre as características clínicas da secreção vaginal e os achados microbiológicos no exame citológico, foi aplicado o índice kappa (k). RESULTADOS: A prevalência da VB, candidíase e tricomoníase foi de 25,5, 9,3 e 2,0%, respectivamente. A sensibilidade, especificidade, valor o VPP e o VPN das características clínicas da secreção vaginal para o diagnóstico citológico de VB foram de 74, 78,6, 54,3, e 89,9%, respectivamente. A sensibilidade, a especificidade, o VPP e o VPN das características clínicas da secreção vaginal para o diagnóstico citológico de candidíase foram de 46,4, 86,2, 25,5 e de 94%, respectivamente. O grau de concordância entre a avaliação clínica da secreção vaginal e o diagnóstico microbiológico de VB, candidíase e tricomoníase, avaliados pelo índice kappa foi de 0,47, 0,23 e 0,28, respectivamente. CONCLUSÃO: A causa mais frequente de secreção vaginal anormal foi VB. A avaliação clínica da secreção vaginal apresentou sensibilidade, VPP e grau de concordância moderado a fraco, comparado ao diagnóstico microbiológico, o que indica a necessidade de avaliação complementar do achado clínico de secreção vaginal anormal.

Introduction Penile carcinoma (PC) is a rare disease, however it is still considered a serious public health problem. The expression of p16INK4a, a protein associated with tumour suppression, can be used as a marker for the presence of high risk HPV DNA. The upregulation of this protein is understood to be an attempt to stop uncontrolled cellular proliferation in response to HPV infection. Objectives The goal of this study was to estimate the prevalence of HPV DNA and evaluate the expression and correlation of p16INK4a with HPV DNA in patients with PC in Goias, Brazil. Methods: this retrospective cohort study involved 190 patients with PC treated in the UroOncology service of Hospital Araujo Jorge (HAJ), a unit of the Association Against Cancer in Goias (ACCG), from January 2003 to November 2015. The paraffin blocks containing the cancerous tissue fragments were subjected to extraction of viral DNA, subsequently subjected to polymerase chain reaction testing with short PCR fragment (SPF PCR) primers to detect HPV DNA. The marking of the p16INK4a protein was performed with immunohistochemistry, using a commercial kit (Mach 4 Universal HRPPolymer Detection System – Biocare Medical, CA, USA). The slides were evaluated independently by two pathologists. Results Of the 190 samples tested, 89 (46.8%) (CI 95%: 39.8%–53.9%) showed positive HPV DNA and 98 (51.7.0%) (CI 95%: 33.2 to 53.2) showed expression of p16INK4a. The correlation between the presence of HPV DNA and p16INK4a was 63.6% (CI 95%: 46.3 to 78.6). Although there is no expression of p16INK4a in 100% of cases positive for HPVDNA, there was statistical significance between the presence of viral DNA and expression of p16INK4a (p Conclusion Some studies suggest that the standard knowledge of the expression of the p16INK4a protein may be a useful marker for HPV activity in patients with penile cancer. The results of this study showed that there are significant differences between the expression of this protein in positive and negative HPV DNA samples.

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.

Cervical cancer, caused by high oncogenic risk Human Papillomavirus (HPV) infection, continues to be a public health problem, mainly in developing countries. Using peptide phage display as a tool to identify potential molecular targets in HPV associated tumors, we identified α-mannosidase, among other enriched sequences. This enzyme is expressed in both tumor and inflammatory compartment of the tumor microenvironment. Several studies in experimental models have shown that its inhibition by swainsonine (SW) led to inhibition of tumor growth and metastasis directly and indirectly, through activation of macrophages and NK cells, promoting anti-tumor activity. Therefore, the aim of this work was to test if swainsonine treatment could modulate anti-tumor immune responses and therefore interfere in HPV associated tumor growth. Validation of our biopanning results showed that cervical tumors, both tumor cells and leukocytes, expressed α-mannosidase. Ex vivo experiments with tumor associated macrophages showed that SW could partially modulate macrophage phenotype, decreasing CCL2 secretion and impairing IL-10 and IL-6 upregulation, which prompted us to proceed to in vivo tests. However, in vivo, SW treatment increased tumor growth. Investigation of the mechanisms leading to this result showed that SW treatment significantly induced the accumulation of myeloid derived suppressor cells in the spleen of tumor bearing mice, which inhibited T cell activation. Our results suggested that SW contributes to cervical cancer progression by favoring proliferation and accumulation of myeloid cells in the spleen, thus exacerbating these tumors systemic effects on the immune system, therefore facilitating tumor growth.

Summary Objectives To determine the frequency of human papillomavirus (HPV) types and behavioral characteristics related to cytological abnormalities in women descendants of slaves, who live in isolated communities known as quilombos in the state of Maranhao, Brazil. Methods Cervicovaginal specimens of 353 women were analyzed by conventional cytology and genotyping. HPV detection and genotyping was performed using a linear array HPV genotyping test kit. Behavioral factors and their association with cytological abnormalities were analyzed, as well as the association between cytological abnormalities and HPV infection. Results The frequency of HPV infection was 13%, and infection with high-risk HPV types was more frequent than with low-risk types (10.2% vs. 2.8%). The most prevalent genotypes were HPV 68 (3.1%) and HPV 58 (2.6%). HPV-positive women were 6.5 times more likely than HPV-negative women to be diagnosed with cytological abnormalities. There was a significant association between HPV infection and the presence of cytological abnormalities in women 31–40 years of age and in women 51–60 years of age. Conclusions A distinct profile of high-risk HPV genotypes was detected, with predominance of types 68 and 58. It is possible that the results of the present study are due to specific characteristics of the population, which is geographically isolated and maintains conservative sexual habits.

Glandular abnormalities comprise less than one-quarter of the diseases affecting the uterine cervix. The acquisition of knowledge on the epidemiological and biological features of glandular lesions of the cervix has occurred more slowly compared with the progress associated with squamous cell lesions. This difference is principally due to the greater prevalence of squamous cells lesions compared with that of glandular lesions. Evidence-based management guidelines are lacking for the most severe forms of glandular abnormalities of the cervix, in particular for invasive adenocarcinomas. This review summarizes the epidemiology, natural history and diagnosis of glandular abnormalities of the cervix and the current debate on the management of cervical in situ and invasive adenocarcinomas.

Cytopathic effects related to the human papillomavirus (HPV) infection are more frequently found in cervical intraepithelial neoplasia (CIN) 1; however, there are indications that at least half the histological diagnoses of CIN2 and CIN3 include koilocytosis areas. The objective of this study was to evaluate the frequency of the cytological criteria suggestive of HPV infection in the cervical smears of women with a histological diagnosis of CIN. One hundred and sixty-two women with abnormal cervical smears and a diagnosis of CIN confirmed by histopathology were selected, including 46 cases of CIN 1, 42 of CIN 2 and 74 cases of CIN 3. Koilocytosis was found in 63% of the smears from women with a histopathological diagnosis of CIN 1. This sign was observed in 26.2% and 25.7% of smears of women with a diagnosis of CIN 2 and CIN 3, respectively. Cytomegaly also was frequent in cervical smears of women with histopathological diagnosis of CIN 1 (71.8%). On the other hand, spindle cells and atypical metaplasia were more frequent in women with CIN 2 and CIN 3. Atypical parakeratosis showed similar frequency in all grades of CIN diagnosis. Koilocytois and cytomegaly were inversely correlated with the diagnosis of CIN2 or CIN 3, with OR values respectively of 0.30 (95%CI 0.13-0.68) and 0.26 (95%CI 0.11-0.58). The others signs analyzed did not show any significant association. Koilocitosis and cytomegaly can provides good reassurance that a patient with atypical cervical smear have CIN 1.

Objective To evaluate the outcome of CIN 2 diagnosed by colposcopy-directed biopsy in women followed without treatment for 12 months and to verify whether the regression and progression of this lesion are associated with the woman's age at diagnosis and age at first sexual intercourse. Study design Women diagnosed with CIN 2 by biopsy and with previous cervical smear showing LSIL were included in this cohort study and followed up for one year with cervical smear and colposcopy every three months. The rates of progression, persistence and regression of the CIN 2 were evaluated. The Kruskal–Wallis test was used to analyze the woman's age at diagnosis, age at first sexual intercourse and interval since the first sexual intercourse according to the CIN 2 outcome, assuming a significance level of 5%. Results At the end of 12 months of follow-up the CIN 2 regression rate was 74% (31/42), progression rate to CIN 3 was 24% (10/42) and in one case CIN 2 persisted (2%). Among women who had regression, this event was detected in the first six months of follow-up in 26 of the 31 cases. There was no statistically significant association between the evolution of CIN 2 and the woman's age at diagnosis, age at first sexual intercourse and interval since first sexual intercourse. Women whose lesions were restricted to one quadrant were more likely to have CIN 2 regression at three-month follow-up compared with women with a lesion extending to one or more quadrants (OR: 6.50; 95% CI: 1.20–35.23). Conclusions The results of this study indicate that the majority of CIN 2 diagnosed by biopsy in women with previous Pap smear showing LSIL will regress in 12 months and therefore an expectant approach could be considered in these cases, not only for young women. Nevertheless these findings are not conclusive, and larger studies are required in order to certify when it is safe to adopt expectant management for CIN 2.

Introducao: a colpocitologia oncotica tem se mostrado um metodo de triagem eficaz na prevencao de câncer de colo uterino e de suas lesoes precursoras. Objetivo: identificar as alteracoes celulares presentes nos resultados da colpocitologia oncotica realizada em reeducandas. Metodos: estudo de corte transversal e analitico, realizado com reeducandas do Complexo Prisional da Regiao Metropolitana de Goiânia, Goias. Resultados: participaram da entrevista 148 mulheres, e 104 realizaram o exame colpocitologico. A populacao constituiu-se por mulheres que possuiam no maximo 30 anos de idade (57,6%), eram naturais de Goias (66,3%), solteiras (45,2%), com baixa escolaridade e baixa renda. Quanto a adequabilidade da amostra, 94,2% foram consideradas satisfatorias. O exame colpocitologico identificou 10,2% de alteracoes celulares classificadas em: ASC-US (4,1%), ASG-US (2,0%), LSIL (1,0%) e HSIL (3,1%). Conclusao: os resultados deste estudo mostram a necessidade de adocao de politicas de saude voltadas para este grupo.

This study was designed to analyze whether specific human papillomavirus (HPV) genotypes may predict histologic outcomes in women with glandular abnormalities in their cervical smears. Of the 160 women included, 111 were diagnosed with atypical glandular cells, 35 had both atypical glandular cells and high-grade squamous intraepithelial lesions, whereas 14 women had AIS, in 1 case associated with high-grade squamous intraepithelial lesions. All women underwent colposcopic examinations and biopsy was performed in 129/160 (80.6%). Thirty-one women (19.3%) were considered negative for neoplasia and scheduled for follow-up. All specimens were tested for 27 HPV genotypes by Roche's polymerase chain reaction-reverse line blot assay. Histologic diagnoses were either cervical intraepithelial neoplasia or invasive carcinoma in 75 (58%) women, and negative for neoplasia in 54 (42%). The overall prevalence of HPV was 43%. HPV 16 was the most prevalent type followed by HPV 18. HPV 16 was significantly associated with squamous and glandular neoplasia and HPV 18 with glandular neoplasia. In women with cervical intraepithelial neoplasia 2 or 3, 11 different HPV genotypes were found, whereas in those who had invasive glandular or invasive carcinoma HPV 16 and HPV 18 were found predominantly. The detection of HPV 16 in women with glandular abnormalities in cervical smears did not help differentiating squamous from glandular lesions. However, the detection of HPV 53 in abnormal smears can predict squamous neoplasia, whereas HPV 18 can predict glandular neoplasia as histologic diagnoses.

The value of repeat cytology in the detection of high-grade preinvasive or invasive disease was analyzed at the time of colposcopy in cases of patients with abnormal referral cytology. A series of 136 patients with squamous abnormalities in their referral cytology, who had undergone repeat cytology and for whom outcome had been established, were included in the study. Final diagnosis consisted of 27 negative results (20%), 37 cases of CIN1 (27%), 31 CIN2 (23%), 39 CIN3 (29%), and 2 (1%) cases of squamous cell carcinoma (SCC). Referral and repeat cytology suggestive of atypical squamous cells cannot exclude high-grade squamous intraepithelial lesion (ASC-H), high-grade squamous intraepithelial lesion (HSIL) were findings significantly associated with a final diagnosis of CIN2 or worse. Sensitivity and positive predictive value of repeat cytology for the histological diagnosis of significant histological neoplasia (CIN2 or worse) were 81.9% and 77.6%, respectively. In women with referral cytology findings of atypical squamous cells of undetermined significance or low-grade squamous intraepithelial lesion (ASC-US/LSIL), repeat cytology was suggestive of HSIL in 20% of cases (11/54), and in 4% (2/54) biopsy was negative or CIN1, and the final diagnosis established by LLETZ was CIN3. Eight cases (10%) of HSIL in referral and repeat cytology in which biopsies were negative or insufficient were diagnosed as CIN3 or worse in the final diagnosis. Repeat cytology would have changed therapeutic management in 4 and 10%, respectively, of patients included as ASC-US/LSIL and ASC-H/HSIL in referral cytology. Diagn. Cytopathol. 2009. © 2008 Wiley-Liss, Inc.

Background The objective of this study was to evaluate the correlation between the 2001 Bethesda classification of endocervical glandular abnormalities and histological diagnosis. Study design A series of 155 women with endocervical glandular abnormalities on cervical smears were included: 91 with atypical glandular cells (AGC) not otherwise specified (NOS), 15 with AGC-favor neoplastic (FN); 35 with AGC associated with high-grade squamous intraepithelial lesion (HSIL) as combined diagnosis and 14 with adenocarcinoma in situ (AIS). Results Histological outcome of squamous neoplasias (CIN 2 or worse) and adenocarcinoma were significantly associated with AGC-FN and AIS, taking as reference AGC-NOS, and more associated with AIS than AGC-FN. Similar associations were observed for histological outcome of adenocarcinoma, but no association was observed for only squamous neoplasia. Histological outcome of CIN2 or worse was strongly associated with AGC when HSIL was also present, but no association was observed with only for adenocarcinoma histological outcome. Conclusions AGC-NOS, AGC-FN and AIS cytological diagnosis represent a progressively increasing association with neoplastic diagnosis, due to progressively increasing association with adenocarcinoma. Histological outcome of squamous neoplasia is frequent but does not differ with these cytological interpretations. The presence of HSIL associated with AGC represents greater probability of squamous neoplasia but not adenocarcinoma.

Objective: To analyse the correlation between cytomorphological criteria in smears with atypical glandular cells (AGC) or adenocarcinoma in situ (AIS) and human papillomavirus (HPV) reflex test results with different neoplastic histological diagnoses, particularly to distinguish between glandular and squamous neoplasia. Methods: A series of 155 women with glandular abnormalities in their conventional cervical smears was included: 106 with AGC, 35 with AGC associated with high-grade squamous intraepithelial lesion (HSIL) and 14 with AIS. Two reviewers evaluated 35 cytomorphological criteria and hybrid capture II (HCII) was performed in all cases. Colposcopy was carried out in all cases and biopsy in 126/155. For statistical purposes, predictive values and odds ratio (OR) were calculated, followed by chi-square automatic interaction detection. Results: Histology detected 56 cases of squamous and 17 of glandular intraepithelial or invasive neoplasia. Predictive values of the papillary groups and feathering criteria for glandular neoplasia were, respectively, 80.0% and 73.3%. Feathering was the criterion with the highest OR for distinguishing glandular from squamous neoplasia and also for distinguishing between glandular and non-neoplastic diagnosis. Rosettes and pseudostratified strips did not perform as well. Multivariant Classification and Regression Trees analysis identified feathering as the best criterion for distinguishing between glandular, squamous and non-neoplastic diagnoses regardless of HPV status. Conclusions: Feathering was the best criterion for predicting glandular neoplasia.

Human papillomavirus (HPV) infection, bacterial vaginosis (BV), and cervicitis may play a role in the presence of cytological abnormalities in female adolescents and young women. This study aimed to estimate the prevalence of HPV, BV and cervicitis in female adolescents and young women and evaluate whether these conditions are associated with a finding of cytological abnormalities in cervical smears.Cervical smears were screened using the conventional method and HPV-DNA detection was performed by PGMY-PCR. BV was defined as ≥20% clue cells in cervical smears. Cervicitis was assessed by counting leukocytes in five non-adjacent microscopic fields at 1000× magnification. Odds ratios and 95% confidence intervals were calculated.The study analyzed 251 samples, with cytological abnormalities being found in 9.5% (24/251). Atypical squamous cells of undetermined significance (50.0%) and low-grade squamous intraepithelial lesions (29.1%) were the most common findings. HPV, BV and cervicitis were found in 44.2% (111/251), 41.0% (103/251) and 83.2% (209/251) of cases, respectively. Of the variables investigated, BV and an abnormal cytological diagnosis were independently associated with HPV positivity. Cytological abnormalities were significantly associated with a finding of HPV and BV in the same woman, and also with a simultaneous finding of HPV, BV and cervicitis; however, the independent association of these combined variables did not remain significant in the multivariate analysis.BV and cytological abnormalities were independently associated with HPV infection in female adolescents and young women.

Background: Co-infection by Chlamydia trachomatis (CT) in women with Human Papillomavirus (HPV) infection has been shown to increase the risk of developing cervical intraepithelial neoplasia (CIN). The present study was designed to analyze the association between HPV e CT co-infection and the severity of cervical neoplasia. Methods: Two hundred fifty-one women with PCR-confirmed HPV infection were tested for CT co-infection by PCR, prior to cervical conizations due to CIN. Prevalence rates of CT and HPV types were reported for the histological diagnosis categories. Results: The prevalence of Chlamydia trachomatis was 15.1% (38/251). CT negative women showed a significant association between age ≥ 30 years and CIN 2 or worse diagnosis; this association was not found in CT positive women. In women < 29 years of age, negative for CT, the infection by HPV 16 /18 were detected in 50% of the women with CIN 2 or worse diagnosis and in 19.5% of women with CIN 1 or negative (OR=5.83; 95%CI: 2.19-15.57). Conclusion: No association with CIN 2 or worse diagnosis was observed for Chlamydia trachomatis positive women for all age groups. These data can suggest that HPV type and no CT infection may correlate with risk for severity of histological diagnoses in younger women.

Aim: To evaluate the expression of MMP-2, MMP-9, MMP-14, TIMP-1 and TIMP-2 in Cervical intraepithelial neoplasia grade 3 (CIN 3) and invasive squamous cell carcinoma. Methods and Results: This study comprised three groups: Group 1: 55 cases with CIN 3; Group 2: 30 cases with CIN 3 and invasive carcinoma components; Group 3: 46 cases with invasive carcinoma. Protein expression was investigated in tumor and stromal cells by immunohistochemistry and percentages of immunostained cells were determined. The mean percentage of MMP-14 tumor cells was significantly higher in stromal cells in all groups. TIMP-2 and MMP-9 expression was significantly higher in stromal cells than in tumor cells. TIMP-1 had a significantly higher expression in stromal cells of carcinoma and tumor cells of CIN 3. Stromal cells expression of MMP-2, MMP- 14, TIMP-1 was increased with the severity of cervical neoplasia. The expression of MMP-2 in stromal cells that was higher in CIN 3 component of Group 2 than in CIN 3 of Group 1. Conclusions: The increase in MMP-2 expression from CIN 3 to invasive cervical cancer reinforces their role in cervical cancer progression. From CIN 3 to invasive cervical carcinoma, TIMP-1 expression increased in stromal cells and decreased in tumor cells.

Angiogenesis seems to be an early event that is associated with disease progression in cervical intraepithelial neoplasia (CIN). The aim of this study was to describe and analyze VEGF-A expression in CIN 3 and invasive cervical carcinoma, in tumor cells and stromal cells. This study comprised three groups: Group 1: 53 cases with CIN 3; Group 2: 24 cases with both CIN 3 and invasive carcinoma components; Group 3: 36 cases with invasive carcinoma. Protein expression was investigated in tumor and stromal cells by immunohistochemistry. Statistical analysis was carried out, considering the mean percentage of immunopositive cells. VEGF-A expression was higher in stromal cells in cases of invasive carcinoma (Group 3) than in CIN 3 cases (Group 1) and this difference was statistically significant. VEGF-A expression showed a borderline association between stromal cells in invasive carcinoma (Group 3) and invasive carcinoma associated with CIN 3 (Group 2). The mean percentage of cells expressing VEGF-A was significantly higher in tumor cells in CIN 3 than in stromal cells in CIN 3 (Group 1). The progression of CIN 3 to invasive carcinoma seems be determined by complex interactions between tumor and stromal cells. These findings reinforce the evidence of the role of stromal cells in tumor invasiveness.

Objective: To evaluate the presence of some criteria in cervical smears with atypical glandular cells and their correlation with histological patterns to identify pre-neoplastic and neoplastic lesions. Methods: Seventy-three women referred with an atypical glandular cell smear, who had undergone conization or hysterectomy, were included in this study. Referral Pap smears were reviewed using the set of 27 cyto-morphological criteria that was correlated with the histological diagnosis. Results: Histological results showed intraepithelial or invasive neoplasia in 35 (48%) cases and benign lesions in 38 (52%) cases. After logistic regression and decision tree analysis an increased nuclear/cytoplasmic ratio and the presence of dyskeratotic cells were strongly associated with intraepithelial or invasive neoplasia and the differential cyto-morphological criteria for glandular lesions were decreased cytoplasm, irregular nuclear membranes and the presence of nucleoli. Conclusion: The analysis of individual cyto-morphological criteria can better predict intraepithelial or invasive neoplasia and differentiate glandular from squamous lesions.

Background and objective: Human Papillomavirus (HPV) persistence after high-grade cervical intra-epithelial neoplasia (CIN) removal may be associated with residual lesions or risk of disease recurrence. Knowledge regarding the factors associated with HPV persistence following CIN treatment is still limited. The main purpose of this longitudinal study was to assess the association between characteristics of the patients and their cervical lesions with high-risk HPV-type persistence, detected by commercially available Hybrid Capture II (HC II), after CIN 2 and 3 treatment with large loop excision of the transformation zone (LLETZ). Study design: For this cohort study, a total of 94 women submitted to LLETZ between March 2001 and September 2002 were included. Only women with at least one follow-up visit at 6 or 12 months and confirmed CIN 2 or 3 in the cone specimen were considered. In each visit women answered to a questionnaire and undertook Pap smear and HC II specimens collection. McNemar’s, chi-square and Fisher tests were used for univariate analysis. Generalized Estimating Equations (GEE) were used for multivariate analysis. All calculations were performed within 95% confidence intervals (95% CI). Results: Histological evaluation showed 12 (13%) women with CIN, 2 and 82 (87%) with CIN 3 and conization margins were compromised in 27 (29%) cases. Eighty-seven (92%) women showed positive HC II tests prior to LLETZ. Of women initially HPV negative, none had a positive HC II during follow-up. The proportion of positive HPV tests was reduced from 92% to 20%(P < 0.01) at the first visit and to 22% (P < 0.01) at the second visit after LLETZ. Multivariate analysis showed that smoking and age above 35 years (irrespective of margin status) were strongly associated with positive HPV during follow-up. Conclusion: HPV persistence following LLETZ was associated with smoking and with the interaction between age and conization margins.

We assessed the detection of high-risk human papillomavirus DNA (HPV-DNA) in women examined by a second Pap smear due to atypical glandular cells (AGC) detected in their screening Pap smear. In 91 women included in the study, a second Pap smear was taken and HPV-DNA test was performed using Hybrid Capture II (HC II). The second Pap smear showed no abnormalities in 28 (31%) cases, ACG in 17 (19%) cases, and high-grade squamous intraepithelial lesions (HSIL) or worse in 24 (26%). HC II test was positive in 36% of the cases. Considering the second Pap smear diagnosis, HPV-DNA was detected in 87% of the women with HSIL, 100% of women with in situ adenocarcinoma, and only in 11% of the women with no abnormalities. The use of the second Pap smear combined with HPV-DNA may improve the management of women with AGC in the primary screening. Diagn. Cytopathol. 2004;31:19–22. © 2004 Wiley-Liss, Inc.

The objective of this study was to analyze the association between bacterial vaginosis (BV) and squamous intraepithelial lesion (SIL). Pap smears were analyzed to verify the presence of BV and SIL. One hundred and ten women with SIL comprised the study group, while 110 women with no cytological abnormalities served as controls. BV was similarly present in women of both groups: 18% of women with SIL and 12% of women without SIL. Results were also similar when the grade of SIL was taken into consideration. BV was detected in 16% of women with low-grade SIL and in 12% of women in the control group, while a higher rate of BV (33%) was found among women with high-grade SIL in comparison to the controls (12%). This difference, however, was not statistically significant. BV tended to be more common among women with high-grade SIL than in women with no cytological abnormalities. Diagn. Cytopathol. 2006; 34:323–325. © 2006 Wiley-Liss, Inc.

Purpose. To describe the acquisition, persistence, and clearance of HPV infection in women with CIN 2 followed up for 12 months.Methods. Thirty-seven women with CIN 2 biopsy, who have proven referral to cervical smear showing low-grade squamous intraepithelial lesions or atypical squamous cells of undetermined significance and tested for HPV, were followed up for one year with cervical smear, colposcopy, and HPV test every three months. HPV DNA was detected by the polymerase chain reaction and genotyping by reverse line blot hybridization assay.Results. CIN 2 regression rate was 49% (18/37), persistence as CIN 1 or CIN 2 was 22% (8/37), and progression to CIN 3 was 29% (11/37). Multiple HPV types were observed at admission in 41% (15/37) of cases. HPV 16 was detected at admission in 58% (11/19) of the cases that persisted/progressed and in 39% (7/18) of the cases that regressed. HPV 16 was considered possibly causal in 67% (10/15) of the cases that persisted or progressed and in 10% (1/10) of the cases that regressed (P=0.01).Conclusion. Multiple HPV infections were frequently detected among women with CIN 2 at admission and during the followup. The CIN 2 associated with HPV 16 was more likely to persist or to progress to CIN 3.

Background Human papillomavirus and Chlamydia trachomatis share the same route of sexual transmission and possess similar risk factors, indicating that coinfection may act synergistically in the induction of epithelial cell abnormalities. Objective This study aimed to determine the prevalence of human papillomavirus and Chlamydia trachomatis in adolescents and young women and identify factors associated with coinfection. Study Design This cross-sectional study included 276 female participants, aged 15–24 years, who were sexually active. Interviews were conducted and cervical specimens were collected for cervical smears and molecular tests. All cervical specimens were tested for 27 human papillomavirus genotypes by polymerase chain reaction amplification and hybridization to a human papillomavirus linear array. Detection of Chlamydia trachomatis was performed by polymerase chain reaction using primers directed to the region encoding the cryptic plasmid. Bivariate and multivariate analyses were performed to evaluate the factors associated with coinfection with human papillomavirus and Chlamydia trachomatis . The odds ratio, the adjusted odds ratio, and the 95% confidence interval were calculated. Results The prevalence of infection by Chlamydia trachomatis and human papillomavirus was 9.1% (95% confidence interval, 5.61–12.4) and 47.1% (95% confidence interval, 41.0–53.2), respectively. The prevalence of coinfection with human papillomavirus and Chlamydia trachomatis was 5.8% (95% confidence interval, 3.3–9.2); coinfection with 1 human papillomavirus type was 3.3% (95% confidence interval, 1.5–6.1) and with multiple types was 2.5% (95% confidence interval, 1.0–5.2). The prevalence of cytological abnormalities was 12.3% (95% confidence interval, 8.6–16.79). Human papillomavirus infections of high oncogenic risk were more prevalent (85.4%). Factors independently associated with coinfection of human papillomavirus/ Chlamydia trachomatis obtained by multivariate analysis were the initiation of sexual activity under 16 years of age with an an odds ratio of 4.9 (95% confidence interval, 1.0–23.63; P = .05) and cytological abnormalities with an odds ratio of 10.7 (95% confidence interval, 1.9–59.5; P = .01), which indicates there is risk for the detection of cytological abnormalities in adolescents and young women coinfected with human papillomavirus/ Chlamydia trachomatis . Conclusion The prevalence of coinfection among our study population was of a magnitude that warrants attention by public health services. Adolescents and young women should be monitored for Chlamydia trachomatis infection and vaccinated against human papillomavirus. The association between cytological abnormalities and coinfection with human papillomavirus and Chlamydia trachomatis indicates the potential synergistic role of these infections in carcinogenesis of the cervix.

High-risk human papillomavirus (HPV) is considered the main etiological agent for cervical neoplasia. However, the presence of a single type HPV infection alone is unlikely to be sufficient to cause cervical cancer. There is epidemiologic evidence suggesting that HPV and Chlamydia trachomatis play a central role in the etiology of cervical intraepithelial neoplasia and subsequent cervical cancer. To evaluate the HPV prevalence and the seropositivity for C. trachomatis in women referred to the colposcopy clinic due to an abnormal cervical smear and to examine the effect of this association on the severity of cervical neoplasia. Following enrollment, 131 patients underwent colposcopy and biopsies when necessary. HPV DNA was detected by the polymerase chain reaction (PCR) and genotyping was performed by reverse line-blot hybridization assay. C. trachomatis seropositivity was tested by ELISA for the detection of IgG antibodies. The prevalence of HPV infection was 86.3%. Seropositivity for C. trachomatis was 26%. Thirty-one women (27.4%) were positive for C. trachomatis antibodies and HPV-DNA. The most prevalent HPV type in C. trachomatis-seropositive women were HPV 16 (51.6%) and this HPV type was present mainly in neoplasia cases. Positivity for HPV, particularly HPV types 16 and 18, and C. trachomatis seropositivity was significantly associated with a diagnosis of high grade neoplasia. Borderline significance was observed after adjustment for HPV. C. trachomatis seropositivity is associated with high grade neoplasia in women infected with HPV, mainly when the types 16 and 18 were involved.

This study was designed to evaluate the effect of single or multiple-human papillomavirus (HPV) infection and phylogenetic groups on the prevalence and severity of cervical intraepithelial neoplasia (CIN) in women undergoing colposcopy after an abnormal cervical smear. Colposcopy was performed in 198 cases and biopsy was performed in 193 patients. All specimens were tested for 27 HPV genotypes using the Roche polymerase chain reaction reverse line blot assay. The overall prevalence of HPV infection in women with an abnormal cervical smear was 86% (171 of 198). The prevalence of HPV 16 in high-grade CIN (2/3) was 52% (40 of 76), being detected in 88.8% of cases (8 of 9) of invasive carcinoma. The prevalence of HPV types 31 and 35 in high-grade CIN was 10.5% (8 of 76) and 6.6% (5 of 76), respectively. Single or multiple-type infection involving HPV 16 were significantly associated with a diagnosis of high-grade neoplasia (≥ CIN 2) [odds ratio (OR) 6.49; 95% confidence interval (CI): 1.88-23.44 and OR: 3.65; 95% CI: 1.13-12.15] even after adjustment for HPV-DNA. A statistically significant association was also found between HPV 16 and the other HPV types belonging to species α 9 and a diagnosis of high-grade neoplasia (OR: 7.62; 95% CI: 1.28-51.58); however, no association was found between HPV 16 and the other HPV types belonging to species α 7. HPV 16 is the most important predictor of high-grade cervical neoplasia. Multiple-type infections are predictors of high-grade cervical neoplasia when type 16 is present.

The objective of this study was to evaluate the frequency and the significance of cytomorphological criteria defined in studies as being predictive of neoplasia in cervical smears of women with a cytological diagnosis of atypical glandular cells (AGC) or adenocarcinoma in situ (AIS). Women (n = 103) with cytological findings suggestive of AGC or AIS, whose diagnoses were later established by histopathology, were included in the study. The criteria analyzed and classified as present or absent in cervical smears previously classified as AGC-NOS (not otherwise specified), AGC-FN (favor neoplasia), or AIS were as follows: irregular nuclear membranes; scanty cytoplasm; dyskeratotic cells; increased nuclear/cytoplasmic ratio; nucleoli; overlapping; papillary clusters, feathering; loss of polarity; nuclear enlargement; coarsely granular chromatin; and pseudostratified strips. Histopathology resulted in neoplastic diagnoses in 55 cases (53.3%) and nonneoplastic diagnoses in 48 cases (46.6%). Coarsely granular chromatin was observed in 62.5% of cases with a diagnosis of neoplasia. Feathering was present in 80% of cases of histopathological AIS. Loss of polarity and coarsely granular chromatin were significantly associated with neoplastic diagnosis considering all subcategories of glandular abnormalities diagnosis. In AGC-SOE subclassification, coarsely granular chromatin was significantly associated with neoplastic diagnosis. The presence of nucleoli was significantly associated with neoplastic diagnosis in cervical smears qualified as AGC-FN and AIS. Nuclear enlargement, increased nuclear/cytoplasmic ratio, coarsely granular chromatin and overlapping cells were found in all the subclassifications of glandular cell abnormalities irrespective of the histopathological results. Chromatin aspects, polarity, and presence of nucleoli can predict neoplasia.

The relation between the detection of clue cells in cervical smears of women with CIN and the expression of COX-2 in these lesions were determined. Samples from 228 women, treated due to CIN and who underwent cervical conization, were obtained. Hybrid Capture II and Pap smear samples were collected immediately before performing conization. Pathological diagnoses were 11 (5%) normal cervix, 35 (15%) CIN1, 31 (14%) CIN2, and 151 (66%) CIN3. COX-2 immunoreactivity grading on the pathological specimens was based on the German ImmunoReactive score. In cervical smears, 20 fields (40x) were examined, each of them with a minimum count of 10 epithelial cells. When 20% or more of clue cells were detected the sample was considered positive for clue cells. The prevalence of clue cells was similar across histological strata (P = 0.42). Although the expression of COX-2 did not differ in lesions with varying severities (P = 0.24), there was a negative association between the expression of COX-2 and the presence of clue cells in Pap smear (OR = 0.4; 95% CI = 0.2-0.9): only 12% of women with moderate and strong expression of COX-2 had clue cells in their smears, contrasted to 22% of those with negative and weak expression of COX-2. HPV infection was associated in a borderline manner to the expression of COX-2 (P = 0.04; OR = 2.3 95% CI = 1.0-5.4). The reduced expression of COX-2 in CIN specimens may suggest that clue cells interfere with the inflammatory component of the carcinogenic process that lead to CIN.

The recovery and stability of DNA for the detection and genotyping of HPV in UCM-containing specimens, after exposure to denaturing reagents and stored for up to 2 years were evaluated. Samples were collected from 60 women who had cervical cytology specimens harboring cervical intraepithelial neoplasia (CIN) 2 or 3. All samples were stored in UCM and had been frozen at −20 °C following the addition of the denaturing reagent (sodium hydroxide) and the removal of the aliquot required for Hybrid Capture 2 testing for the identification of HPV DNA. The samples had been stored for 6, 12 and 24 months (20 samples for each storage time). HPV DNA extraction was performed according to a protocol designed specifically and the presence and quality of DNA was confirmed by human β-globin detection using the consensus primers G73 and G74. HPV DNA was amplified using the consensus primers PGMY09 and PGMY11, and reverse line-blot hybridization was used to detect type-specific amplicons for 37 HPV types. The DNA extracted from the denatured specimen was recovered in 57/60 (95%) of the samples. HPV DNA was detected in 56/57 (98%) of the recovered samples. Twenty-six of the 56 samples recovered (48%) were genotyped successfully.