Carol Morin

Human papillomavirus and cancer of the uterine cervix

Abstract Human papillomavirus infection of the cervix was found in 1.69% of 234,715 women during a mass screening program during the years 1975 to 1979 inclusive. Condylomata of the cervix represent the most prevalent squamous lesion diagnosed during this 5-year period. Most of condylomata (71.16%) were found in women under 30 years of age with a mean of 27.47 years and a peak incidence of 2.5% in the 20–25 year-age group. Condylomata precede invasive carcinoma by 27.45 years. The coexistence of condylomata was observed in 25.6% of dysplastic and neoplastic lesions. This association seems to shorten the mean time for malignant transformation. The epidemiologic behavior of condylomata appears similar to dysplasia which is considered a precursor of neoplasia. However, the high regression rate of condylomata (68.39%) implies that lowered immunity and other promoting or synergistic factors are necessary for malignant conversion.

Insulin-Like Growth Factor-I, IGF-Binding Protein-3, and Mammographic Breast Density

Some studies have suggested that insulin-like growth factor (IGF) pathway is related to premenopausal breast density, one of the strongest known breast cancer risk factors. This study was designed specifically to test the hypothesis that higher levels of IGF-I and lower levels of IGF-binding protein (IGFBP)-3 are associated with high mammographic breast density among premenopausal but not among postmenopausal women. A total of 783 premenopausal and 791 postmenopausal healthy women were recruited during screening mammography examinations. Blood samples were collected at the time of mammography, and plasma IGF-I and IGFBP-3 levels were measured by ELISA. Mammographic breast density was estimated using a computer-assisted method. Spearmans partial correlation coefficients (rs) were used to evaluate the associations. Adjusted mean breast density was assessed by joint levels of IGF-I and IGFBP-3 using generalized linear models. Among premenopausal women, high levels of IGF-I and low levels of IGFBP-3 were independently correlated with high breast density (rs = 0.083; P = 0.021 and rs = −0.124; P = 0.0005, respectively). Correlation of IGF-I with breast density was stronger among women in the lowest tertile of IGFBP-3 than among those in the highest tertile of IGFBP-3 (rs = 0.138; P = 0.027 and rs = −0.039; P = 0.530, respectively). In contrast, the correlation of IGFBP-3 with breast density was stronger among women in the highest tertile of IGF-I than among those in the lowest tertile of IGF-I (rs = −0.150; P = 0.016 and rs = −0.008; P = 0.904, respectively). Women in the combined top tertile of IGF-I and bottom tertile of IGFBP-3 had higher mean breast density than those in the combined bottom tertile of IGF-I and top tertile of IGFBP-3 (53.8% versus 40.9%; P = 0.014). No significant association was observed among postmenopausal women. Our findings confirm that IGF-I and IGFBP-3 are associated with breast density among premenopausal women. They provide additional support for the idea that, among premenopausal women, these growth factors may affect breast cancer risk, at least in part, through their influence on breast tissue morphology as reflected on mammogram.

Human papillomaviruses and vulvar vestibulitis

Objective To assess the relationship between human papillomavirus (HPV) infection and vulvar vestibulitis syndrome. Methods From November 1995 to December 1997, 135 women with vulvar vestibulitis were compared with 322 controls who had no evidence of vulvar vestibulitis. Human papillomavirus DNA was amplified by polymerase chain reaction and detected with liquid-capture molecular assay. Results Human papillomavirus DNA was found in 29.6% of cases and in 23.9% of controls (relative risk [RR] 1.4; 95% confidence interval [CI] .8, 2.2). The prevalence of HPV tended to decrease with increasing duration of pain among cases. Thus, prevalences were 37.5%, 29.6%, and 22.0% for pain durations of 3–6 months, 7–12 months, and 13–24 months, respectively (P = .14). Prevalence of HPV also tended to increase with pain intensity among cases, but that association was not statistically significant (P = .57). Prevalence percentages for women with low, moderate, or severe pain were 27.5%, 28.8%, and 34.4%, respectively. Prevalence of HPV was slightly higher in cases with the most severe pain (34.4%) than in controls (23.9%) (RR 1.8; 95% CI .8, 4.0). In cases with the most pain in the shortest time (3–6 months), prevalence of HPV was double that of controls (50% versus 23.9%) (RR 3.5; 95% CI 1.0, 12.7; P = .054). Conclusion There was little support for the idea that HPV might be related to vulvar vestibulitis.

Human papillomavirus and cervical lesions.

Human papillomavirus (HPV) infection has been implicated in the intraepithelial cervical changes that cause most abnormal Papanicolaou smears. To date, 14 types of HPVs have been identified. All are small, nondeveloped, icosahedral DNA viruses that share a common internal antigen. In cases of cervical HPV infection, the koilocytes and dyskeratocytes are the most frequently seen cell types. Most infections are flat aceto-white lesions. Florid condyloma acuminatum, usually detectable with the naked eye, is characterized by an irregular surface secondary to finger-like projections, in the middle of which a capillary loop comes to the surface. Spiked condyloma, not seen with the naked eye, has an irregular surface that shows asperity. 3 techniques have been used to differentiate atypical condyloma from intraepithelial neoplasia: microspectrophotometric studies, the peroxidase- antiperoxidase technique, and electronmicroscopy. There is growing evidence that papillomaviruses play an etiologic role in human genital cancer. 20-25% of dysplastic and neoplastic lesions show a coexistence of condylomas of the cervix or vulva with dysplasia or neoplasia. Epidemiologic research suggests that cervical condylomas occur at a mean age of 27.5 years, precede cervical dysplasia by 3.3 years, carcinoma in situ by 9.3 years, and invasive carcinoma by 27.4 years. The conversion of most cases of papillomas into squamous cell carcinomas requires the presence of carcinogenic initiators, 1 of which is believed to be herpes simplex virus.

VAGINAL CONDYLOMATA: A HUMAN PAPILLOMAVIRUS INFECTION

In our clinic, as a rule, we do not treat vaginal condylomata. They are usually subclinical and asymptomatic. When atypia is present on biopsy, they should be treated in the same manner as vaginal intraepithelial neoplasia. When vaginal discharge and pruritus are present, infection should be searched for and treated. When condylomata are seen with the naked eye, colposcopy has shown that there were many more, too small to be seen, so that local therapy seems a waste of time. If on colposcopic examination only a few condyloma acuminata are located, then therapy is defendable. CO2 laser therapy should be preferred to other modalities until a systemic treatment is available and safe.

Quantitative DNA methylation analysis of laser capture microdissected formalin-fixed and paraffin-embedded tissues

We developed an assay to quantify DNA methylation in breast cancer cells isolated by laser capture microdissection (LCM). The assay uses methylation sensitive restriction enzyme (MSRE) digestion and quantitative polymerase chain reaction (qPCR). To assess the validity and precision of the assay, we prepared standard samples with expected methylation percentage (MP) for two gene promoters (PLAU (plasminogen inhibitor, urokinase) and TIMP3 (TIMP metallopeptidase inhibitor 3)) that we compared with measured MPs. We found good linearity of MSRE digestion and qPCR procedures for both promoters (beta=0.90-1.19+/-0.05-0.10 and r=0.95-0.98; all P<0.0001). Moreover, results remained similar after addition of a purification step between MSRE digestion and qPCR procedures. The validity of this technique was also confirmed by successfully replicating previously published MPs of four cell lines for PLAU and TIMP3 promoters. We assessed the consistency of our approach by comparing MPs of PLAU and TIMP3 promoters from nine breast cancer patients and two cell lines using LCM frozen tissues and their corresponding formalin-fixed paraffin-embedded tissues. We found good consistency (intraclass correlation coefficient=0.93) of MPs between frozen tissues and formalin-fixed paraffin-embedded tissues. Our data demonstrate that this assay based on digestion with MSRE and qPCR procedures is a good technique to quantify MP on limited amounts of DNA and may find clinical applications.

Cytologic Predictors of Cervical Intraepithelial Neoplasia in Women with an ASCUS Pap Smear

OBJECTIVE To identify cytologic parameters on Pap smears of women with an atypical squamous cells of undetermined significance (ASCUS) diagnosis that could help cytologists to indicate whether a particular ASCUS case is most likely related to cervical intraepithelial neoplasia (CIN) grade 1 or 2/3. STUDY DESIGN A total of 360 eligible women diagnosed with ASCUS and referred to the colposcopy clinic of Saint-Sacrement Hospital participated in the study. Eligible women were those aged 18-50 years, newly diagnosed with ASCUS, with no history of cervical biopsies or treatment, and not pregnant at the time of the visit. Colposcopically directed biopsies of lesions were obtained. All Pap smears were reviewed according to 36 different cytomorphologic criteria. The regression logistic model was used to estimate the odds ratios (ORs) for the associations between cytologic criteria observed in smears and the diagnosis of CIN made on biopsies. All cytologic criteria significantly (P < .05) associated with CIN were entered in the models, and a backward selection was done to determine independent cytologic predictors of CIN 1 and 2/3. RESULTS Biopsies revealed that 22.2% of the study population had concurrent CIN. CIN I and 2/3 were identified in 61 (16.9%) and 19 women (5.3%), respectively. Clear perinuclear spaces (OR = 2.5, P = .002) and moderate nuclear atypia (OR = 4.4, P = .02) were two cytologic criteria independently associated with CIN 1. Four independent predictors of CIN 2/3 were identified: the presence of clear perinuclear spaces (OR = 5.9, P = .004), hyperchromasia (OR = 3.9, P = .04), moderate anisokaryosis (OR = 13.1, P = .01 and increased nuclear volume of metaplastic cells (OR = 5.1, P = .007). CONCLUSION These observations may help cytologists to better categorize ASCUS lesions as intraepithelial ones and will also contribute to improving the Bethesda definition of ASCUS. Further studies are planned to validate these observations.

Evaluation of a Modified Sanitary Napkin as a Sample Self-Collection Device for the Detection of Genital Chlamydial Infection in Women

ABSTRACT A modified sanitary napkin was compared with endocervical swab and urine specimens for the detection of urogenital Chlamydia trachomatis infection. Endocervical swabs and/or first-catch urine were collected from 510 women at medical or community settings in Quebec City. Participants were also asked to wear a modified sanitary napkin (Ezy-Detek) during 4 consecutive hours and to bring it back to the clinic or mail it to the laboratory. Endocervical and urine specimens were tested using the Cobas Amplicor CT/NG assay (Roche Diagnostic Systems) according to the manufacturers instructions, as were specimens collected with the napkin after adequate preparation. If the PCR test result was positive on the endocervical sample or on any two samples, a woman was considered to be infected. PCR testing results on paired samples were identical for 493 (96.6%) of 510 women. According to the definition given above, 58 (11.3%; 95% confidence interval [CI], 8.7 to 14.5%) women were infected with C. trachomatis. The sensitivity and specificity of PCR testing on modified sanitary napkin specimens were, respectively, 93.1% (54 of 58; 95% CI, 83.3 to 98.1%) and 98.9% (447 of 452; 95% CI, 97.4 to 99.6%) compared to 81.0% (47 of 58; 95% CI, 68.6 to 90.1%) and 100% (451 of 451; 95% CI, 99.2 to 100%) for urine specimens. The positive and negative predictive values were, respectively, 91.5% (54 of 59) and 99.1% (447 of 451) for the sanitary napkin specimens compared to 100% (47 of 47) and 97.6% (451 of 462) for urine samples. These results suggest that a modified sanitary napkin represents an effective noninvasive device for self-collection of specimens to detect urogenital C. trachomatis infection.

Immunohistochemistry of breast tumor markers on archived bouin-fixed paraffin-embedded tissues.

Neutral-buffered formalin is the most commonly used tissue fixative in pathology laboratory. Among other fixatives, Bouins solution has been used in several laboratories and is still in use for particular tissues. In this project, we determine if we can study breast clinical markers on archived Bouin-fixed tissue samples with immunohistochemistry (IHC) protocols optimized for tissue fixed in neutral-buffered formalin. To evaluate the concordance of IHC results between formalin-fixed and Bouin-fixed tissues, we calculated the concordance percentage and the &kgr; statistic of 12 clinical IHC markers quantified by an automated system on breast cancer tissues fixed in neutral-buffered formalin and their corresponding tissues fixed in Bouins solution. When positivity threshold of immunostaining was setup at ≥10% for both fixation conditions, we observed a concordance percentage of 83.9% (&kgr;=0.65). However, when positivity threshold of immunostaining was lowered to 3% to 4% for Bouin-fixed tissues, concordance percentage was then of 96.8% (&kgr;=0.92). Our data demonstrate that we can study IHC markers on archived Bouin-fixed tissue from patients with long clinical follow-up using IHC protocols optimized for formalin-fixed tissues after an adjustment of the positivity threshold of immunostaining quantified by an automated system.

Cervical condyloma planum.

Abstract Condylomata were defined and described as wart-like lesions with papillary projections often observed on the skin and mucosa of the anogenital area. Cytologic, histologic, and colposcopic studies have demonstrated that the condylomatous lesion on the cervix presents itself more frequently as a flat lesion than a classical exophytic papillary one. Condyloma planum represents a new lesion on the human cervix, not described until 1977. 1,2 The viral etiology of this new type of condyloma has been confirmed in three ways: by the electron microscopic observation of human papillomavirus (HPV) particles, 1,3–8 by the identification of viral antigen using the peroxidaseantiperoxidase technique, 9–13 and by demonstrating the presence of the HPV DNA sequences using molecular cloning and hybridization techniques. 5,14