High-grade Squamous Intraepithelial Lesion Cells Research Articles

Effects of Menstrual Cycle on Various Morphologies of High-Grade Squamous Intraepithelial Lesions in SurePath™ Liquid-Based Cervical Cytology

Introduction: The morphology of high-grade squamous intraepithelial lesion (HSIL) on Papanicolaou (Pap) smears widely varied, including syncytial aggregates, sheets, and scattered single cells, and no particular cellular pattern is consistently observed. Therefore, this study aimed to determine whether the menstrual cycle affects the cellular pattern of HSILs, an effort to avoid false negatives due to the oversight of scattered small single HSIL cells in the cytological triage of human papillomavirus-positive women. Methods: A total of 147 HSIL samples of liquid-based cytology (LBC) in patients with cervical intraepithelial neoplasia grade 2 or 3 were obtained, and then, the relationship between cellular patterns, such as single-cell-like and syncytial aggregates, and menstrual cycles classified into six phases was analyzed. If a syncytial aggregate was present, the number of cells constituting the aggregate was visually counted under the microscope. Results: HSILs in scattered single cells and small sheets of <6 on LBC samples accounted for 43% (23/54) during the late proliferative phase of the menstrual cycle. A moderately strong statistically significant association was observed between cellular patterns and menstrual cycles (χ² [3] = 9.423, p < 0.05) (Cramer’s V = 0.253). The value of adjusted residuals showed a statistically significant increased proportion of single-cell-like patterns during the late proliferative phase (p < 0.01). Conclusions: The present study demonstrated that HSIL cells in Pap smears in the late proliferation phase have a high frequency of single-cell-like patterns. In human papillomavirus-positive Pap smears with a clean background and predominantly superficial cells, careful microscopic observation by targeting single HSIL cells can potentially reduce false negatives.

Performance of specific morphologic features in distinguishing low-grade squamous intraepithelial lesions from high-grade squamous intraepithelial lesions in borderline cases: a College of American Pathologists Cytopathology Committee multiobserver study.

Distinguishing between low-grade squamous intraepithelial lesions (LSIL) and high-grade squamous intraepithelial lesions (HSIL) can be difficult on certain Papanicolaou (Pap) tests, hindering interobserver concordance. We investigated the variables influencing the interpretation of LSIL versus HSIL in Pap test slides rejected from the College of American Pathologists PAP education program. Eleven cytologists, who were unaware of the reference interpretation, examined 21 Pap slides (11 submitted as LSIL and 10 as HSIL) rejected from the PAP education program and recorded the number of LSIL cells, HSIL cells, keratinized dysplastic cells, LSIL clusters with mixed HSIL cells, atypical squamous metaplasia, atypical glandular cells, the presence of inflammation or infectious organisms, and the overall interpretation (LSIL or HSIL). We evaluated the significance of these 11 variables using a nonlinear mixed model analysis. LSIL had greater concordance (92 of 121 responses; 76.0% concordance) than HSIL (68 of 110 responses; 61.8% concordance; P < 0.001). The only predictors of misclassified cases were the number of atypical squamous metaplastic cells and the number of HSIL cells (P < 0.001). The more of these cells identified, the more likely the reviewers were to classify the slide as HSIL. The reproducibility of the diagnosis was fair (Gwet's agreement coefficient, 0.33). Interobserver reproducibility is a challenge for a subset of cases with features intermediate between LSIL and HSIL. Atypical squamous metaplasia and dysplastic nuclei with a nuclear/cytoplasmic ratio greater than one half of the cell volume (HSIL) present on a Pap test influenced the likelihood that a reviewer would interpret the case as HSIL rather than LSIL.

Treatment Outcomes of Patients With Cervical Intraepithelial Neoplasia or Invasive Carcinoma Who Underwent Loop Electrosurgical Excision Procedure.

BackgroundThis study aimed to evaluate the treatment outcomes of cervical intraepithelial neoplasia (CIN) or cancer patients who underwent loop electrosurgical excision procedure (LEEP) in terms of primary outcome and factors associated with persistence/recurrence.MethodsPatients with CIN or cancer who underwent LEEP from January 2007 to December 2015 were reviewed. Data collected were age, parity, menopausal status, human immunodeficiency virus (HIV) infection, smoking, cervical cytology, histopathology from cervical biopsy and LEEP including margin status, final histopathology, and follow-up data.ResultsThe mean age of 385 patients was 41.9 ± 10.8 years (range 18 - 79 years). Majority were multiparous (81.6%) and premenopausal (78.2%). There were 15.3% of patients with HIV infection. The most common cervical cytology was high-grade squamous cell intraepithelial lesion (HSIL, 44.1%), followed by atypical squamous cells of undetermined significance (ACS-US, 21%). Minor complications of bleeding or infection from LEEP were encountered in 7.3%. Among 153 patients (39.7%) who had positive margin(s), 43 underwent second LEEP, whereas 76 had hysterectomy. From all patients, 47 had failure after treatment (12.2%), being either persistence (30 patients; 7.8%) or recurrence (17 patients; 4.4%). Factors associated with persistence or recurrence by multivariate analysis were age ≥ 55 years old, HIV infection, final diagnosis of invasive cancer, and positive endocervical margin or both ecto- and endo- cervical margins.ConclusionsLEEP had low rate of persistence/recurrence. Age ≥ 55 years old, HIV infection, final diagnosis of cancer, and positive endocervical or both endo- and ecto- surgical margin(s) were significantly associated with persistent or recurrent diseases.

Aptima HPV messenger RNA testing and histopathologic follow‐up in women with HSIL cytology: A study emphasizing additional review of HPV‐negative cases

High-risk human papillomavirus (hrHPV) messenger RNA (mRNA) testing, the Food and Drug Administration-approved testing platform since 2013, has been increasing as a cervical screening alternative to hrHPV DNA testing methods. This study reports the largest routine clinical follow-up study reported to date of hrHPV mRNA cotesting and histopathologic follow-up results for women with high-grade squamous intraepithelial lesion (HSIL) cytology results. HSIL Papanicolaou test results for women cotested with Aptima hrHPV mRNA testing between June 2015 and November 2020 were analyzed along with recorded histopathologic follow-up results within 6 months of screening. Aptima hrHPV mRNA-positive results were reported for 95.2% of the cotested HSIL cytology cases (905 of 951). Histopathologic cervical intraepithelial neoplasia grade 2 or worse (CIN2+) was diagnosed on follow-up in 538 of 701 hrHPV mRNA-positive cases (76.8%) and in 15 of 36 hrHPV mRNA-negative cases (41.7%). Additional reviews of the hrHPV mRNA-negative HSIL cases showed variable interpretations, and confirmatory blinded-review interpretations of HSIL or atypical squamous cells, cannot exclude high-grade squamous intraepithelial lesion were more likely in cases with histopathologic CIN2+ (77.5% [93 of 120]) than those with cervical intraepithelial neoplasia grade 1 or negative findings (63.1% [101 of 160]; P < .01). This large routine-clinical-practice study confirms the previously reported high sensitivity of hrHPV mRNA testing for the detection of high-grade cervical dysplasia and cervical cancers. The blinded-review findings indicate that additional cytology review may be helpful for confirming an interpretation of HSIL in daily practice, especially for hrHPV-negative HSIL cases.

Screening for intra-anal squamous intra-epithelial lesions in women with a history of human papillomavirus-related vulvar or perianal disease: results of a screening protocol.

Women with a history of human papillomavirus (HPV)-related cervical, vaginal or vulvar high-grade squamous intra-epithelial lesions (HSILs) or cancer are at increased risk of developing anal squamous intra-epithelial lesions (SILs) or a squamous cell carcinoma of the anus (SCCA). Screening for intra-anal SILs with high-resolution anoscopy (HRA) in high-risk populations is a subject of debate. In this study we aimed to answer the following question: what is the prevalence of intra-anal (H)SIL in women with HPV-related vulvar and/or perianal disease using HRA for screening? A retrospective study was performed to evaluate the prevalence of intra-anal (H)SIL in women with a history of vulvar and/or perianal HSIL or (superficially invasive) squamous cell carcinoma (SCC). This study was performed between 2015 and 2018 following implementation of a protocol for intra-anal screening using HRA. Twenty-seven patients, 10 with a history of (superficially invasive) SCC (four vulvar, five perianal, one multizonal) and 17 with HSIL as the worst diagnosis (two perianal, 15 multizonal) were screened for intra-anal lesions using HRA. No anal cancer was found at screening, 6 (22%) patients were diagnosed with intra-anal HSIL and 12 (44%) patients with intra-anal low-grade SIL. We found a high prevalence of intra-anal HSIL in women previously diagnosed with vulvar and perianal HSIL. Given the clear link between HSIL and SCCA, screening for intra-anal lesions in women with HPV-related genital pathology seems warranted. Future studies should focus on the effect of HSIL treatment on the prevention of anal cancer.

Significant decrease of a master regulator of genes (REST/NRSF) in high-grade squamous intraepithelial lesion and cervical cancer

BackgroundThe repressor element 1-silencing transcription factor (REST) is a regulator of gene expression, and the Ras association domain family member 1 A (RASSF1A) is an important tumor suppressor gene involved in cancer development. Although extensive characterization of the roles of REST and RASSF1A in cancer development have been reported in cellular models, the link between them and their possible role in the development of squamous intraepithelial lesions (SIL) and squamous cell carcinoma (SCC) of the cervix have not been explored. The aim of this study was to evaluate the expression of REST and RASSF1A in cervical cytological and histological samples from patients diagnosed with SIL or SCC and in CC-derived cell lines. MethodsWe analyzed the expression of REST and RASSF1A by immunocyto/histochemistry in cervical samples from patients (n = 271) and in cancer cell lines. Data analyses were performed using the Kruskal–Wallis test and generalized linear models. ResultsWe identified binding sites for REST in RASSF1A and observed a significant reduction in REST and RASSF1A nuclear expression in samples from patients with high-grade SIL (HSIL) and SCC. For REST, we observed an average decrease of 334 and 423 r.u.d. for HSIL (n = 21) and SCC (n = 18) compared with non-LSIL (n = 72), whereas for RASSF1A, this decrease was 126 and 217 r.u.d., respectively (p < 0.001). ConclusionOur results provide evidence of the altered expression of REST and RASSF1A in SIL and SCC, with a significant decrease in HSIL, SCC, and SCC-derived cell lines; findings that can contribute to the diagnosis, prognosis, and post-treatment follow-up of patients diagnosed with SIL or SCC.

The interpretation of high-grade squamous intraepithelial lesion on anal cytology: a comparative analysis with the cervical Papanicolaou test

Prior studies have shown that high-grade squamous intraepithelial lesion (HSIL) tends to be underdiagnosed on anal cytology. Our study aims to decipher the interpretative challenges of HSIL that are more specific to anal cytology specimens by comparing them to cervical Papanicolaou tests. One hundred cases each of anal and cervical cytology specimens with HSIL interpretation and concordant histologic follow-up were retrieved and diagnostically confirmed. Patient demographic data were obtained from the electronic medical record. The cytologic specimens were reviewed and statistically compared in terms of proportion of HSIL cells, HSIL patterns and types, and cytoplasmic area of HSIL cells (with digital image analysis). A P value of <0.05 was considered statistically significant. Of the patients with anal HSIL, 97% were human immunodeficiency virus-positive and 60% were men who have sex with men. The anal cytology specimens significantly differed from the cervical ones in several respects: proportion of HSIL cells, cytoplasmic area of HSIL cells, cases with HSIL cells in syncytial groups (10 versus 57) and cases with keratinizing HSIL (45 versus 10). The P value was <0.0001 for all comparisons except for the proportion of HSIL cells (P = 0.001). Anal cytologic HSIL, in contrast to its cervical counterpart, exhibits fewer abnormal cells and smaller size of the diagnostic cells with a higher percentage of keratinizing lesions. A careful scrutiny of the sample with an enhanced understanding of the morphology and better sampling may help improve the detection of anal HSIL on cytology.

Endocervical glandular involvement is associated with an increased detection rate of high-grade squamous intraepithelial lesions on the Papanicolaou test

Although The Bethesda System for Reporting Cervical Cytopathology does not mandate reporting of endocervical glandular involvement (EGI) in Papanicolaou test specimens with high-grade squamous intraepithelial lesions (HSIL), several studies have suggested that EGI diagnosed on surgical specimens is associated with higher rates of residual or recurrent dysplasia. When suspected, EGI is reported for Papanicolaou test specimens at our institution, but the performance of this diagnosis has not been assessed. The archives were queried for Papanicolaou test specimens with a diagnosis of HSIL-EGI (2006-2017). All follow-up surgical pathology specimens within a year of the Papanicolaou test diagnosis were evaluated for cytologic-histologic correlation. This same query was repeated for all surgical pathology specimens with a diagnosis of HSIL-EGI. All preceding Papanicolaou test diagnoses within a year were assessed for cytologic-histologic correlation. Twenty Papanicolaou test specimen glass slides were reviewed by 6 observers to assess for interobserver variability. Patients with HSIL-EGI on surgical specimens were more likely to have a preceding Papanicolaou diagnosis of HSIL and atypical squamous cells, cannot exclude high-grade squamous intraepithelial lesion (ASC-H) (32.3% versus 25.5%, P = 0.03, and 16.7% versus 11.8%, P = 0.04, respectively). Patients with an HSIL-EGI diagnosis on a Papanicolaou test were significantly more likely to have HSIL-EGI detected on a follow-up histology (41.6% versus 24.0%, P < 0.001). Interobserver concordance was poor for the assignment of EGI in Papanicolaou test specimens. Overall, the diagnosis of HSIL-EGI on Papanicolaou test specimens is complicated by poor sensitivity and interobserver concordance.

Reappraisal of cytology-histology correlation in cervical cytology based on the recent American Society of Cytopathology guidelines (2017) at a cancer research centre.

To assess the impact of recently published American Society of Cytopathology (ASC) guidelines (2017) on the conduct of cervical cytology-histology correlation (CHC). A retrospective review was conducted for cervical biopsies with their corresponding conventional cervical smears over a 7.5-year period (January 2011-June 2018). As per the ASC guidelines, a discrepancy assessment grid was prepared. Major cytology-histology discordance was defined as a diagnosis of high-grade squamous intraepithelial lesion (HSIL) or CIN2+ in one of the tests with negative result in the other. Smears and biopsies of all discordant cases were reviewed for reasons of overcall and undercall. Of the 341 cervical biopsies with corresponding Papanicolaou smear, cytology-histology agreement was noted in 249 (73%) cases. Major discordance was observed in 22 cases (6.4%)-16 undercalls and six overcalls on cytology-while minor discrepancies were noted in 70 cases. Atypical metaplasia and repair changes were the main reasons for overcall while small HSIL cells in atrophic smear and scant HSIL cells were important causes of undercall on cytology review. Using the ASC guidelines, we could improvise upon the existing CHC methodology for categorisation of cyto-histological pairs of cases with a cytological diagnosis of atypical glandular cells. The present study demonstrates, for the first time, that the recent ASC guidelines facilitate cervical CHC, especially for categorisation of cases with atypical glandular cells on cytology. Uniform application of these guidelines would standardise the conduct of cervical CHC internationally and provide scope for inter-laboratory comparison of data as well as enhance self-learning and peer learning.

Cervical high-grade squamous intraepithelial lesion on conventional cytology: Cytological patterns, pitfalls, and diagnostic clues.

Conventional cervical cytology, which has a relatively low sensitivity in diagnosing high-grade lesions as compared to liquid-based cytology, is still being practiced in low resource settings. This study aimed at elucidating various cytomorphological patterns, pitfalls, and subtle clues to high-grade squamous intraepithelial lesion (HSIL) diagnosis on conventional cervical cytology through cytologic-histologic correlation. Cervical biopsies reported as CIN2/3 were correlated with their corresponding Pap smears over a 10-year period to determine the frequency of undercalls. For characterization of overcalls, cervical smears reported as HSIL and their corresponding biopsies during the same period were correlated. The discordant cases in both the groups were reviewed for problematic patterns and pitfalls in cytological diagnosis of HSIL. Of the 142 biopsies with CIN2/3, 29 (20.4%) cases had been undercalled on cytology. Sixteen (16) of these could be reclassified as ASC-H/HSIL on smear review. Smears showing predominant cells of low-grade squamous intraepithelial lesion grade with a few HSIL cells and those with small abnormal cells in an atrophic background formed the main confounders for HSIL underdiagnosis. Thirteen (13) out of 130 (10%) Pap smears called as HSIL, where biopsy diagnosis was less than CIN2, were labeled as overcalls. Atypical metaplasia, hyperchromatic crowded groups, and reparative changes constituted the major diagnostic pitfalls on cytology. A diligent smear review helped to reduce the undercall and overcall rates to 9.1% and 2.3%, respectively. Awareness of morphological challenges in interpretation of HSIL among cytopathologists practicing cervical cytology would assist in reducing the diagnostic errors and ensure better patient management.

Perception of Nursing Students towards Palliative Care of Cervical Cancer in Sudan

Objective: To assess the visual inspection with acetic acid (VIA) and Lugol's iodine (VILI) as alternative screening methods for cancer cervix. Materials and methods: Comparative cross-sectional study was conducted on 1000 women with age range from 18 to 61 years were attending the obstetrics and gynecology Department in Zagazig university hospital from January2013 to October 2015. Each one was done Papanicolaou smear (PAP), visual inspection with 5% acetic acid (VIA) and with 5% Lugol’s iodine (VILI). All women underwent Colposcopy. Analyse of the sensitivity, specificity and predictive values of the results using colposcopic directed biopsy as reference was done. Results: From 80 positive screening tests by (either PAP, VIA, VILI or colposcopy). Pap smear was positive in 14/80 (17.5%), including 4 cases of atypical squamous cell with undetermined significance (ASCU), 4 cases of low grade squamous cell intraepithelial lesion (LSIL) and 5 cases of high grade squamous cell intraepithelial lesion (HSIL) and one case with malignant cells. Biopsy was positive in 11/14 of PAP smear. VIA accounted positive in 23/80 (28.7%) and VILI results were positive in 12/80. Biopsy was positive in 21/23 for VIA and 8/12 was positive for VILI , The Pap smear had a sensitivity of 78.57%, specificity of 96.75%, and positive predictive value 75.12% and negative predictive value of 97.09%. VIA had a sensitivity of 91.30%, specificity of 85.33% and positive predictive value of 40.11% and negative predictive value of 98.05%. VILI had sensitivity of 66.54, specificity of 91.32, and positive predictive value 43.51% and negative predictive value of 98.31%. Conclusion: Precancerous and cancerous lesions of the cervix could be well detected by VIA and VILI in low resource locality. Using both testes in matching improves the specificity of both to make them good options for screening of cancer cervix in this society.

Prevalence of high-grade anal dysplasia among women with high-grade lower genital tract dysplasia or cancer: Results of a pilot study

ObjectiveTo estimate the prevalence of high-grade anal dysplasia in women with high-grade dysplasia or carcinoma of the cervix, vagina or vulva. MethodsIn this cross-sectional study, participants underwent anal cytology, anal HPV testing with Cervista HPV16/18 and high-resolution anoscopy (HRA). Patients with HSIL (high-grade squamous cell intraepithelial lesion) or greater on anal cytology or anal biopsy were referred to a colorectal surgery specialist for further evaluation. ResultsSeventy-five women were enrolled in the study, including 47 with cervical (cervix group), 10 with vaginal (vagina group), 15 with vulvar (vulva group), 1 with cervical and vaginal, and 2 with vulvar and vaginal disease. The median age in the cervix group (40 years (range 26–69)) was substantially younger than in the vagina (60 years (38–69)) and the vulva (59 years (36–75)) groups. Anal HSIL based on composite endpoints of the most severe cytology or histology result was diagnosed in 6 patients (8.0%). Anal cytology revealed HSIL in 2 (2.7%), atypical squamous cells of undetermined significance (ASCUS) in 12 (16.0%), low-grade squamous cell intraepithelial lesion (LSIL) in 2 (2.7%), and was normal in 59 (78.7%) patients. Anal HPV16/18 test was positive in 15 (20.0%), negative in 48 (64.0%) and insufficient in 12 (16.0%) patients. Of the 6 women with high-grade anal dysplasia, three (50%) had a positive anal HPV16/18 test. No case of anal cancer was observed. ConclusionOur results suggest that the prevalence of anal HSIL is elevated among women with HPV-related lower genital tract dysplasia or cancer. To further support the inclusion of this high-risk group into screening guidelines for anal dysplasia, further studies are necessary to determine what screening strategy is suited to this population.

TET1 promotes 5hmC-dependent stemness, and inhibits a 5hmC-independent epithelial-mesenchymal transition, in cervical precancerous lesions

DNA hypermethylation is a driving force in carcinogenesis. However, the role of active DNA hypomethylation in cancer remains largely unknown. This process, facilitated by ten-eleven translocation methylcytosine dioxygenase 1 (TET1), which oxidizes 5-methylcytosine (5 mC) to 5-hydroxymethylcytosine (5hmC), has never been studied in cervical cancer. Here, we found that TET1 and 5hmC correlative increases from normal cervix to Low-grade squamous intraepithelial lesion (LSIL), maximizing in High-grade squamous intraepithelial lesion (HSIL), and decreasing in invasive cancer. Full-length HPV-immortalized HSIL cells demonstrated higher TET1/5hmC levels, and stemness properties, compared to invasive cancer cells. TET1 silencing promoted the epithelial-mesenchymal transition (EMT), to transform precancerous cells in vivo. TET1 increased 5hmC in the ZEB1 and VIM promoters, surprisingly, silencing both genes. TET1 interaction with the histone modifiers, LSD1 and EZH2, on the ZEB1 promoter, resulted in gene silencing, via loss of histone H3K4 trimethylation, and gain of histone H3K27 trimethylation. Taken together, TET1 promotes stemness properties, and inhibits EMT, in HSIL cells, through 5hmC-dependent and -independent mechanisms.

Screening for cervical abnormalities associated with EBV, HPV and HSV-2 infections in South-West Nigeria: A tale between sex and non-sex workers

This study investigated socio-economic and clinical correlates of cervical abnormalities and some viral infections such as Human Papilloma virus (HPV), Epstein-Barr virus (EBV) and Herpes Simplex virus type 2 (HSV-2) among commercial sex workers (CSWs) and non-sex workers (NSWs) of reproductive age. It also estimated serum levels of SCCA1, p16 and Ki67 as potential non-invasive biomarkers of cervical abnormalities. This cross-sectional study included 203 consenting participants (NSWs = 98 and CSWs = 105) within the age-range of 20–49 years. Interview based risk factor questionnaire was administered, Pap smears were made from cervical scrapings and classified accordingly while serum levels of SCCA1, Ki67 and p16, EBV, HPV and HSV2 antibodies were determined by ELISA method. Multivariate and multiple logistic regression analysis were on generated data carried out and significance set at p < 0.05. Among NSWs, higher prevalence of cervicitis, atypical squamous cell of undetermined significance (ASCUS), low grade squamous cell intraepithelial lesion (LSIL) and high grade squamous cell intraepithelial lesion (HSIL) were 18.4%, 16.3%, 11.2% and 8.2%, respectively while the prevalence of the cervical abnormalities were 7.6%, 7.6%, 8.6% and 3.8%, respectively among CSWs (p = 0.04). Decreasing and increasing sero-prevalence of viral infections were observed from cervicitis, ASCUS, LSIL to HSIL among NSWs (77, 56, 54.5 and 50%) and CSWs (75, 87.5, 100 and 100%), respectively (p = 0.08). When only ASCUS, LSIL and HSIL were considered, statistics showed p16 sensitivity and specificity of 64.7% and 79.5%, respectively (Odd ratio {OR} 0.7118), SCCA1 sensitivity and specificity of 83.8% and 97%, respectively (OR 166.9) and ki67 sensitivity and specificity of 63.2% and 94.0%, respectively (OR 26.83). It revealed that serum levels of SCCA1, ki67 and p16 were sensitivity to the classes of Pap smear at p = 0.00; p = 0.00 and p = 0.01, respectively. This study suggests that while HPV infection could independently could promote the development of cervical abnormalities among CSWs, EBV infection and other factors may be potential factors among NSWs. It also suggests that serum levels of SCCA1, ki67 and p16 could be used as effective Pap smear complementary diagnostic biomarkers.

Impact of Implementing 100% Rapid Review as a Quality Control Tool in Cervical Cytology

Objective: We aimed to evaluate the impact of implementing 100% rapid review (100% RR) as a quality control tool in cervical smear cytology. Design: A cross-sectional study was conducted in which cytology findings, false-negative results, and quality indicators were evaluated. The variables were analyzed in 2004, the year in which 100% RR was implemented, and again in 2013, i.e., 10 years on. Results: Detection of atypical squamous cells of undetermined significance (ASC-US) increased from 0.90% in 2004 to 2.47% in 2013 by routine screening. Detection of atypical squamous cells, cannot exclude a high-grade squamous intraepithelial lesion (ASC-H) increased from 13.33% in 2004 to 36.27% in 2013 by 100% RR. Detection of high-grade squamous intraepithelial lesion (HSIL) increased from 0.64% in 2004 to 4.29% in 2013 by routine screening, and increased from 0% in 2004 to 19.61% in 2013 by 100% RR. There was a significant increase (p = 0.00001) in the identification of false-negative results of ASC-US, low-grade squamous intraepithelial lesion, ASC-H, HSIL, and atypical glandular cells. All quality indicators had increased in 2013 when compared to 2004 results. Conclusions: Ten years after the implementation of the 100% RR method, improvements were found in the detection of atypia and precursor lesions and in quality indicators.

Pattern of Cervical Cytology and High Risk Human Papillomavirus Strains in Non HIV Positive Women Presenting for Cervical Cancer Screening in Port Harcourt, Nigeria

Background: The high risk human papilloma virus (hrHPV) is largely associated with cervical cancer with identifiable dysplastic changes on cytology. The use of cervical cancer screening strategies has largely improved the general outcome of cervical malignancies globally. Objective: To compare cervical cytological features against human papilloma virus genotypes in non HIV positive patients presenting for cervical cancer screening in UPTH. Method: A cross sectional prospective study of 82 non HIV positive women who presented for cervical cancer screening at the University of Port Harcourt Teaching hospital. A prestructured questionnaire was used to obtain socio demographic information. Cytobrush extracts of cervical samples were subjected to cytology and human papilloma virus DNA genotyping using real-time polymerase chain reaction. The information obtained was analyzed using SSPP version 20 and results presented as frequency tables, percentages and tested for significance using student t-test and chi square with p value Results: The mean age of the women was 36.61 ± 7.75 with an age range of 22 - 58 years. The prevalence of HrHPV was 4.88%. Twenty-two of the respondent have abnormal cytological pattern. While the 4 cases of HrHPV positivity were of normal cytological pattern and occurred in respondents that were in their reproductive age, the 22 that showed abnormal cervical cytology were mainly of low grade squamous intraepithelial lesion (LGSIL). There was no case of high grade squamous cell intraepithelial lesion (HGSIL). Conclusion: HrHPV prevalence rate of 4.88% occurring in low socioeconomic class patients. All the cervical abnormalies were of LGSIL, hence the risk of transformation into cervical cancer is equally low. There may be no correlation between hrHPV and cervical cytology in low risk patients.

Low CD4 cells and viral co-infection increase the risk of VaIN: Use of SCCA1 and Ki67 as diagno-prognostic biomarkers

This study evaluated the correlation of SCCA1, Ki67 and CD4 cell expressions and classified vaginal smears in individuals co-infected with Human immunodeficiency virus (HIV), Herpes simplex virus 2 (HSV2), Epstein Barr virus (EBV) and Human Papilloma virus (HPV). This crossectional study included 173 participants within the age range of 20–70 years. Vaginal smears were stained by Papanicolaou technique and classified into high-grade squamous cell intraepithelial lesion (HSIL), low-grade squamous intraepithelial lesion (LSIL), atypical squamous cells of undetermined significance (ASCUS) and negative for intraepithelial lesion (NIL). Presence of immunoglobulin M and G antibodies for EBV, HIV, HPV and HSV2, and SCCA1 and Ki67 antigens were determined by ELISA method. Result showed that biomarkers SCCA1 had higher sensitivity (87.5%) to vaginal lesions when compared with Ki67 which had a sensitivity of 70.8% (p > .01). Assays revealed viral co-infections of 96.0% and 16.8% in smears positive and negative for vaginal lesions, respectively (p < .01) with HIV, HSV2 and EBV as the most prevalent type of co-infection (36%). The findings of this study suggest that low CD4 cells and viral co-infection could increase the risk of developing vaginal lesions. This study also suggests that SCCA1 and Ki67 could be used as diagnostic and prognostic biomarkers for vaginal intraepithelial neoplasia (VaIN).

Anal High-Grade Squamous Intraepithelial Lesions in Human Immunodeficiency Virus-Infected Men: A Study of 100 Cases With Emphasis on Cytohistologic Correlation.

Anorectal cytology (ARC) is a widely used screening tool for anal cancer in human immunodeficiency virus (HIV)-infected men who have sex with men (MSM). Its diagnostic accuracy needs to be improved, especially for high-grade squamous intraepithelial lesions (HSILs). Using 100 HIV+ MSM with biopsy-proven anal HSILs, we correlated histologic/cytologic findings. Upon review, HSIL cells were present in 58 cytology samples and absent in 42. Positive samples were higher in cellularity and contained transformation zones ( P < .05). Cytology was able to predict HSILs in 36%, 48%, 68%, and 78% of patients with one, two, three, and four or more high-grade lesions. HSIL cells were identified in all cytology samples initially reported as HSILs or atypical squamous cells, cannot exclude HSIL and in 34 samples reported as low-grade squamous intraepithelial lesions or less. Notably, among this last category, 15 (44%) were keratinized-type HSILs. Our findings should improve the ARC detection rate for anal HSILs, helping to implement ARC as the primary screening tool for anal cancer.

Viral load and integration status of HPV58 associated with cervical lesion severity in women of Northeast China.

We aimed to evaluate the viral load and integration status of human papillomavirus 58 in women with different grades of cervical lesions to determine whether viral load and integration status are related to malignant transformation in HPV58-infected women. A total of 212 cervical specimens were collected from women in Northeast China who had undergone human papillomavirus genotyping and were HPV58-positive. The HPV58 viral load was determined using real-time polymerase chain reaction, and the integration status was discriminated using the ratio of HPV58 E2 gene copy number to E6 gene copy number. The median HPV58 viral load in women with normal cervix or cervicitis, low-grade squamous cell intraepithelial lesion, high-grade squamous cell intraepithelial lesion and cervical cancer was 352.12, 864.21, 1199.75 and 693.04 copies/genome, respectively. High significance was obtained when comparing the viral load of infected women presenting normal/cervicitis with that of the women either with precancerous cervical lesions or cervical cancer (P < 0.05). The HPV58 genome was in the episomal form in 35 samples (16.5%), mixed episomal and integrated forms in 165 (77.8%) samples, and completely integrated into the host genome in 12 (5.7%) samples. The HPV58 E2/E6 copy number ratio in the cervical cancer group was significantly lower than that in the other groups (P < 0.01). The HPV58 viral load in patients with precancerous cervical lesions or cervical cancer increases significantly with disease progression. The HPV58 E2/E6 copy number ratio in patients with cervical cancer is lower than that for less severe cervical lesions, suggesting a high degree of viral integration may be a considerable risk factor for cervical cancer.

Abstract 433: Utility of p16 immunohistochemistry in evaluating negative cervical biopsies following high-risk pap test results

Abstract The Lower Anogenital Squamous Terminology (LAST) Standardization Project for HPV-associated Lesions recommends the use of p16 immunohistochemistry as an adjunct to morphologic assessment of cervical biopsies interpreted as negative from patients that are at high risk for missed high-grade disease (defined as a prior cytologic interpretation of HSIL, ASC-H, ASC-US/HPV-16+, or AGC-NOS) (Darragh et al., 2012). However, few studies have specifically evaluated the utility of performing p16 on negative cervical biopsies and endocervical curettage specimens following high-risk Pap test results. A search of the Stanford Cytopathology database from 7/1/2002-6/30/2012 for cervical cytology cases with diagnoses of ASC-H, HSIL, AGC-NOS, and atypical endocervical cells NOS (AGC-EC) yielded 1517 cases. Of these, 703 cases had histologic follow-up. Biopsies were excluded if diagnosed as atypical, low grade squamous intraepithelial lesion (LSIL), high grade squamous intraepithelial lesion (HSIL), or insufficient, or if the time to follow up was greater than 1 year. Immunostain for p16 (CINtec) was performed on 350 cases from 339 patients (age 16-85 yrs, average 42+/-14) and scored as positive (diffuse strong), negative or equivocal. 6 cases were excluded due to insufficient tissue and 3 cases with equivocal staining were excluded from further analysis. For p16(+) and equivocal cases, HPV in situ hybridization (ISH) (Ventana HPV III Family 16 probe) was performed and the corresponding H&amp;E section reviewed. Based on the H&amp;E review, the diagnosis was revised. Follow-up data was also obtained. 12/341 cases (3.5%) were p16(+) corresponding to missed diagnoses of LSIL (1 cases), HSIL-CIN2 (2 cases), and SIL-indeterminate grade (6 cases). Two biopsies from one patient at different time points exhibited p16 staining of bland metaplastic cells undermining endocervical glands that morphologically did not meet criteria for SIL and were interpreted as atypical metaplasia. Follow-up biopsies showed HSIL. p16 immunostain increases the detection rate of SIL by 3.5% in benign appearing cervical biopsies and endocervical curettages from patients with a prior high-risk Pap test result. The benefit of p16 immunostain is highest for cases with a prior Pap diagnosis of HSIL. Cytoplasmic only staining and staining of surface mucosa, endocervical glands, and tubal metaplasia may lead to over-interpretation of p16 as positive. Reference: Darragh, T. M. et al. The Lower Anogenital Squamous Terminology Standardization Project for HPV-Associated Lesions: background and consensus recommendations from the College of American Pathologists and the American Society for Colposcopy and Cervical Pathology. Arch. Pathol. Lab. Med. 136, 1266-1297 (2012). Citation Format: Alana F. Shain, Shirley Kwok, Ann K. Folkins, Christina S. Kong. Utility of p16 immunohistochemistry in evaluating negative cervical biopsies following high-risk pap test results. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 433.