Kari Syrjänen

Evaluation of visual inspection with acetic acid (VIA), Lugol's iodine (VILI), cervical cytology and HPV testing as cervical screening tools in Latin America: This report refers to partial results from the LAMS (Latin AMerican Screening) study

Objectives: To assess the performance indicators of visual inspection with acetic acid (VIA) and visual inspection with Lugols iodine (VILI) in four Latin American centres participating in the ongoing Latin AMerican Screening (LAMS) study, in settings with moderate incidence of cervical disease and with poorly to moderately well-organized cervical cancer screening. Setting: Three Brazilian centres (São Paulo, Campinas and Porto Alegre) and one Argentine centre (Buenos Aires) recruited a total of 11,834 healthy women to undergo VIA, VILI, conventional Pap smear and Hybrid Capture II (HCII). Methods: Women who had a positive result from any of these tests were subjected to colposcopy and biopsies (if necessary), and women with high-grade cervical intraepithelial neoplasia (CIN) were properly treated. To control for verification bias, 5% of women with normal tests were referred for colposcopy, as were 20% of HCII-negative women. Results: Data on VIA (n=11,834), VILI (n=2994), conventional Pap smear (n=10,138) and HCII (n=4195) were available for test comparisons, calculating sensitivity, specificity, and positive and negative predictive values. Overall test positivity was 11.6% for VIA, 23.0% for VILI, 2.2% for Pap smear (LSIL threshold), 1.1% for Pap smear (HSIL threshold) and 17.1% for HCII. VIA was positive in 61.8% of the women with CIN 1, 57.0% of those with CIN 2, 35.0% of women with CIN 3 and in 21 of 28 (75%) of women with cancer. Approximately 10% of women with no detectable disease had an abnormal VIA. Regarding VILI, 83.3% of women diagnosed with CIN 1 and 62.5% of those with CIN 3 had an abnormal test. VILI failed to detect one of three cases of cancer. Both the sensitivity, specificity and positive predictive value of VIA and VILI in detecting CIN 2 or CIN 3 could be significantly improved depending on the combination with Pap smear or HCII (sensitivity up to 100.0% and specificity up to 99.8%). Conclusions: The LAMS study failed to reproduce the performance figures obtained with VIA and VILI (as stand-alone tests) in some other settings, where the prevalence of cervical disease was higher. However, a combined use of VIA or VILI with the Pap test or HCII allowed specific detection of cervical abnormalities.

Epidermal growth factor receptor, c-erbB-2 proto-oncogene and estrogen receptor expression in human papillomavirus lesions of the uterine cervix

The expression of epidermal growth factor receptor (EGF-R), c-erbB-2 proto-oncogene, and estrogen receptor (ER) was studied immunohistochemically in a series of 97 human papillomavirus (HPV) lesions of the uterine cervix, with special emphasis on their association with the HPV type, grade of intraepithelial neoplasia (CIN), and the natural history of the disease. EGF-R expression was found in 95 of 97 (98%) specimens, mainly in basal and parabasal cells. Diffuse nuclear and cytoplasmic staining was detected in 36 of 97 (37%) samples, of which 29 were HPV positive. This staining pattern was most prominent in HPV 18-positive and in CIN lesions. Weak or moderate c-erbB-2 expression was found in 26 of 97 (27%) specimens. Estrogen receptor expression was observed in 28 of 77 (36%) samples, epithelial staining was seen in 11 of 77 (14%), and stromal staining occurred in 24 of 77 (31%) specimens. No clear-cut associations were established between the EGF-R, c-erbB-2, or ER expression and HPV type, nor in CIN or the clinical course of HPV infections. This failure for EGF-R, c-erbB-2, and ER to be associated with the specific HPV types, grade of CIN, or the clinical course of cervical HPV lesions suggests that the assessment of these factors is of limited value in explaining the development of HPV-associated CIN and in predicting the prognosis of this disease.

Transitional Cell Carcinoma Of The Bladder: Failure To Demonstrate Human Papillomavirus Deoxyribonucleic Acid By In Situ Hybridization And Polymerase Chain Reaction

Human papillomaviruses have been implicated in the pathogenesis of a variety of malignancies, particularly those of the anogenital tract. Some recent reports on the presence of human papillomavirus in bladder cancer have raised the possibility that it might be involved in the development of this malignancy as well. To study this concept, a series of 108 transitional cell carcinomas of the bladder were screened for the presence of human papillomavirus deoxyribonucleic acid (DNA) by in situ hybridization with biotin-labeled human papillomavirus cocktail probe and polymerase chain reaction with human papillomavirus L1 consensus primers. Although the positive controls showed strong hybridization signals, no evidence for human papillomavirus DNA was found in any of the bladder carcinomas by in situ hybridization. Similarly, despite the amplification of a 450 bp product in cervical human papillomavirus lesions (used as positive controls), no signals were obtained in any of the bladder tumors studied. beta-globin gene sequences (110 bp), serving as internal controls, were consistently amplified from all tumor samples, suggesting that cellular DNAs from the carcinoma specimens were sufficient for the amplification reaction. These data indicate that human papillomavirus infection is rare in transitional cell carcinoma of the bladder. The significance of these findings is discussed in relation to previous reports on human papillomavirus involvement in bladder carcinomas.

Polymerase Chain Reaction Amplification of Human Papillomavirus DNA from Archival, Papanicolaou-Stained Cervical Smears

OBJECTIVEnTo test the applicability of four protocols in recovering DNA suitable for amplification with the polymerase chain reaction (PCR) in archival, Papanicolaou-stained cervical smears.nnnSTUDY DESIGNnThe most efficient method was used to isolate DNA from 11 archival, Papanicolaou-stained smears with cytopathic changes due to human papillomavirus (HPV) infection to confirm the presence of HPV DNA.nnnRESULTSnbeta-Globin was successfully amplified in all smears, while HPV DNA was detected in 6 of 11. Four of the four HPV DNA-negative smears were classified as high grade squamous intraepithelial lesions. Failure to detect HPV DNA might have been due to the low copy number of HPV DNA or deletion of the L1 region.nnnCONCLUSIONnHigh cellularity and the method of recovering DNA from the smear are important determinants of successful amplification of HPV DNA in archival cervical smears.

PLA2 (group IIA phospholipase A2) as a prognostic determinant in stage II colorectal carcinoma

BACKGROUNDnApproximately 30% of all colorectal cancer (CRC) patients are diagnosed with stage II disease. Adjuvant therapy is not widely recommended. However, it is well established that a subgroup of patients with stage II are at high risk for recurrence within their lifetime and should be considered for adjuvant chemotherapy. The present work was designed to assess the value of group IIA phospholipase A2 (PLA2) as a predictor of disease outcome in stage II CRC patients with long-term follow-up.nnnPATIENTS AND METHODSnThe present study comprises a series of 116 patients who underwent bowel resection for stage II CRC during 1981-1990 at Turku University Hospital. Archival paraffin-embedded CRC tissue samples were used to prepare tissue microarray blocks for immunohistochemical staining with PLA2.nnnRESULTSnFifty-five percent of all tumors were positive for PLA2. There was no significant correlation between PLA2 expression and age, sex, depth of invasion and lymph node status. In Kaplan-Meier survival analysis, there was a significant (P = 0.010) difference in disease-free survival (DFS) between patients with negative tumors (longer DFS) and those with positive tumors. The same was true with disease-specific survival (DSS), patients with PLA2-negative tumors living significantly longer (P = 0.025). In multivariate (Cox) survival analysis, however, PLA2 was not an independent predictor of DFS or DSS. In subgroup analysis, the right-sided tumors with negative PLA2 staining had remarkably better prognosis (P = 0.010) than PLA2-positive left-sided tumors.nnnCONCLUSIONSnQuantification of PLA2 expression seems to provide valuable prognostic information in stage II CRC, particularly in selecting the patients at high risk for recurrent disease who might benefit from adjuvant therapy.

Over-expression of topoisomerase IIalpha is related to the grade of cervical intraepithelial neoplasia (CIN) and high-risk human papillomavirus (HPV), but does not predict prognosis in cervical cancer or HPV clearance after cone treatment.

Objective: One of the pathways leading to cervical cancer is a loss of normal cell cycle control. Topoisomerase II&agr; and II&bgr; are important nuclear proteins controlling the G2/M checkpoint, and shown to be over-expressed in many human cancers. Their links to oncogenic human papillomavirus (HPV) types and their prognostic value in cervical cancer are practically unexplored. Material and Methods As part of our HPV-PathogenISS study, a series of 150 squamous cell carcinomas (SCC) and 152 CIN lesions were examined using immunohistochemical (IHC) staining for topoisomerase II&agr; (topo II&agr;), and tested for HPV using PCR with three primer sets (MY09/11, GP5+/GP6+, SPF). Follow-up data were available from all SCC patients, and 67 CIN lesions had been monitored with serial PCR for HPV clearance/persistence after cone treatment. Results: Topo II&agr; expression increased with increasing grade of CIN (p = 0.0001), with the most dramatic up-regulation upon progression from CIN2 to CIN3 and peaking in SCC (OR 16.23; 95%CI 7.89-33.38). Topo II&agr; up-regulation was also significantly associated with HR-HPV detection in univariate analysis (OR = 3.07; 95%CI 1.70-5.52), but was confounded by the histological grade (Mantel-Haenszel common OR = 1.622; 95%CI 0.782-3.365), and by entering both p16INK4a (9) and Survivin (33) in the multivariate regression model. Topo II&agr; did not predict clearance/persistence of HR-HPV after treatment of CIN, and it was not a prognostic factor in cervical cancer in either univariate or multivariate analysis. Conclusions: Over-expression of topo II&agr; is significantly associated with progression from CIN2 to CIN3, being a late marker of cell proliferation. Its close association with HR-HPV is plausibly explained by the fact that E7 oncoproteins of these HR-HPV (but not LR-HPV) block the normal pRb-mediated inhibition of topo II&agr; by degrading the wild-type Rb.

Associação entre idade ao início da atividade sexual e subseqüente infecção por papilomavírus humano : resultados de um programa de rastreamento brasileiro

PuRPOse : to investigate women’s age at their first sexual intercourse and its correlation with their present age, human papillomavirus (HPV) infection and cytological abnormalities at Pap smear. MethOds : women from the general population were invited to be screened for cervical cancer and pre-malignant lesions. After answering a behavior questionnaire, they were submitted to screening with cervical cytology and high-risk HPV testing with Hybrid Capture 2 (HC2). This report is part of the Latin American Screening (LAMS) study, that comprises centers from Brazil and Argentina, and the data presented herein refer to the Brazilian women evaluated at the cities of Porto Alegre, Sao Paulo and Campinas. Results : from 8,649 women that answered the questionnaire, 8,641 reported previous sexual activity and were included in this analysis. The mean age at the interview was 38.1±11.0 years and the mean age at the first sexual intercourse was

Variants of the long control region of human papillomavirus type 16.

Expression of the human papillomavirus (HPV) E6 and E7 oncogenes is regulated on the transcriptional level by specific protein-binding sites contained in the viral long control region (LCR). Sequence changes within the LCR region may have an impact on the transcription of viral oncogenes, possibly resulting in differences in the oncogenic potential of the virus. The present study was designed to determine the sequence variability of the LCR of HPV 16 and to assess whether certain LCR variants do correlate with the clinical outcome of the disease of the uterine cervix. The entire LCR segment of HPV 16 was analysed from 37 cervical biopsy specimens derived from 28 women included in the Kuopio long-term prospective follow-up study. The LCR sequence was identical with the reference sequence in six HPV 16 isolates. Overall, 14 different HPV 16 LCR variants were identified. One of the variants showed sequence variation typical of the Asian-American variant lineage of HPV 16, and all the other variants appeared to belong to the European variant group. The European variants exhibited low genetic diversity, and only five of these LCR variants contained nucleotide changes involving known or proposed binding sites for transcription factors. The variants with changes at nucleotide positions 7193 and 7521 was the most prevalent, accounting for almost 37% of infections. This variant (7193; 7521) has been previously demonstrated to have similar transcriptional activity compared with the reference isolate by Veress and colleagues J Gen Virol 1999, 80, 1035-1043. The reference isolate, variant (7193; 7521) and variants with changes within transcription factor binding sites accounted for most of the infections, and no significant differences were found in the comparison of the distribution of these different LCR isolates in cases where the disease showed progression to severe cervical intraepithelial neoplasia (CIN) or carcinoma in situ (CIS). Notably, both the reference isolate and variant (7193; 7521) were also closely associated with infections showing more aggressive behaviour. According to the present findings, in European HPV 16 isolates, intratype genetic variation of the LCR region does not seem to be commonly responsible for differences in the pathogenicity of the virus and thereby for a risk of progressive infections.

Loss of MUC2 expression predicts disease recurrence and poor outcome in colorectal carcinoma

Clinical staging and histological grading after surgery have been the “gold standard” for predicting prognosis and planning for adjuvant therapy of colorectal cancer (CRC). With the recent development of molecular markers, it has become possible to characterize tumors at the molecular level. This is important for stage II and III CRCs, in which clinicopathological features do not accurately predict heterogeneity, e.g., in their tumor response to adjuvant therapy. In the present study, archival samples from 141 patients with stage I, II, III, or IV CRC treated during 1981–1990 at Turku University Hospital (Finland) were used (as microarray blocks) to analyze MUC2 expression by immunohistochemistry. Altogether, 49.7xa0% of all tumors were positive for MUC2. There was no significant correlation between MUC2 expression and age (Pu2009<u20090.499), tumor invasion (Pu2009<u20090.127), tumor staging (Pu2009<u20090.470), histological grade (Pu2009<u20090.706), lymph node involvement (Pu2009<u20090.854), or tumor metastasis (Pu2009<u20090.586). However, loss of MUC2 expression was significantly associated with disease recurrence (Pu2009<u20090.031), tumor localization (Pu2009<u20090.048), and with borderline significance with gender (Pu2009<u20090.085). In univariate (Kaplan–Meier) survival analysis, positive MUC2 significantly predicted longer disease-free survival (DFS) and disease-specific survival (DSS) as well. However, in multivariate (Cox) survival analysis, MUC2 lost its power as an independent predictor of DFS and DSS. Our results implicate the value of MUC2 expression in predicting disease recurrence and long-term survival in CRC.

Safety of screening with Human papillomavirus testing for cervical cancer at three-year intervals in a high-risk population: experience from the LAMS study*

Objectives To assess whether human papillomavirus (HPV) testing is a safe enough approach to warrant extension of the screening intervals of baseline Papanicolaou (Pap)−/HPV− women in low-income settings. Methods Of the >1000 women prospectively followed up as part of the Latin American Screening (LAMS) Study in São Paulo, Campinas, Porto Alegre) and Buenos Aires, 470 women with both baseline cytology and Hybrid Capture 2 (HC2) results available were included in this analysis. These baseline Pap-negative and HC2− or HC2+ women were controlled at six-month intervals with colposcopy, HC2 and Pap to assess the cumulative risk of incident Pap smear abnormalities and their predictive factors. Results Of the 470 women, 324 (68.9%) were high-risk HPV (hrHPV) positive and 146 (31.1%) were negative. Having two or more lifetime sex partners (odds ratio [OR] = 2.63; 95% CI 1.70–3.51) and women using hormonal contraception (OR = 2.21; 95% CI 1.40–3.51) were at increased risk for baseline hrHPV infection. Baseline hrHPV+ women had a significantly increased risk of incident abnormal Pap smears during the follow-up. Survival curves deviate from each other starting at month 24 onwards, when hrHPV+ women start rapidly accumulating incident Pap smear abnormalities, including atypical squamous cells (ASC) or worse (log-rank; P < 0.001), low-grade squamous intraepithelial lesions (LSIL) or worse (P < 0.001) and high-grade squamous intraepithelial lesions (HSIL) (P = 0.03). Among the baseline hrHPV− women, the acquisition of incident hrHPV during the follow-up period significantly increased the risk of incident cytological abnormalities (hazard ratio = 3.5; 95% CI 1.1–11.7). Conclusion These data implicate that HPV testing for hrHPV types might be a safe enough approach to warrant extension of the screening interval of hrHPV−/Pap−women even in low-resource settings. Although some women will inevitably contract hrHPV, the process to develop HSIL will be long enough to enable their detection at the next screening round (e.g. after three years).