Low cost technology for screening early cancerous lesions of oral cavity in rural settings.

BACKGROUND: Low cost technologies are needed in resource poor rural settings for detection of pre-cancer and cancer lesions of the oral cavity. AIM: The study was undertaken to investigate the feasibility of a low cost technology (Magnivisualizer) for the early detection any lesions of the oral cavity among tobacco users in a resource poor rural field setting. SUBJECTS AND METHODS: A total of 1329 tobacco users were motivated to come forward for oral examination in the camp organized for this purpose. Their oral cavities were screened with a torch and Magnivisualizer by a Dentist. RESULTS: With torch light, 104/1329 (7.8%) lesions were identified, though only 62/104 (59.6%) positive lesions could be differentiated into various categories. However, through Magnivisualizer 156/1329 (11.7%) lesions were detected and 153/156 (98.1%) positive lesions were differentiated into different categories. CONCLUSION: Magnivisualizer offers an alternative instrument for detecting most of the early cancerous and high-grade precancerous lesions and it can be used in the rural field settings.

VIA screening for cervical cancer in developing countries: potential role of the light source.

A total of 385 symptomatic patients presenting to the gynaecology and obstetrics outpatient department were screened by two sources of light: the yellow light of the tungsten bulb and the white light of the halogen bulb (100 Watt, 12 Volt), fitted with KODAK 80B filter and diffuser in an instrument called a 'Magnivisualizer' (developed by our Institute). Colposcopic examination was the gold standard for visualisation of the cervix. This study clearly brings out the significance of visual examination of the cervix using white light; as, in addition to having perfect correlation with colposcopy (0.86 for white vs 0.53 for yellow light), white light enables us to select the correct site of biopsy. Most of the rural clinics use torch or ordinary tungsten bulb, thus missing many significant lesions. In the light of our research findings, we strongly recommend the use of white light (complete spectrum of light) for screening purposes.

Urine HPV-DNA detection for cervical cancer screening: prospects and prejudices.

Oncogenic types of human papilloma viruses (HPVs) have been established to be the causative agents for cervical cancers and high-grade squamous intraepithelial lesions (HSILs). The clinical application of molecular tests for HPV detection for screening purposes has been of considerable interest. DNA amplification methods allow the use of self-collected samples (including urine) from material collected away from the original disease site. For screening of cervical pathology, detection of HPV-DNA in urine would be useful only if it represents cervical HPV infection and/or HPV-related cervical pathology. We conducted a review of the literature in order to ascertain: (1) if urine is an adequate sample for HPV-detection; (2) whether sensitive techniques are available for HPV-detection in urine and (3) if detection of HPV in urine truly represents cervical infection/pathology. The review process consisted of assembling facts and analysing the published literature on the following facts: anatomical considerations of the lower genital and the lower urinary tract, biological behaviour of HPV and its shedding behaviour, technical issues regarding sample collection, processing and HPV-DNA assay systems, concordance rates of HPV-DNA detection and their type specificity in the paired samples (urine and cervical scrapes) obtained in different clinico-epidemiological settings and comparative detection rates of HSILs in the paired samples.

Test characteristics of various screening modalities for cervical cancer: a feasibility study to develop an alternative strategy for resource-limited settings.

OBJECTIVE: To determine test characteristics - sensitivity, specificity, positive and negative predictive values - of different screening modalities to detect cervical precancerous and cancerous lesions in order to devise an effective alternative strategy for cervical cancer screening in resource-poor settings. METHODS: A total of 472 women presenting with nonspecific gynecologic symptoms were screened by cytology, visual inspection with acetic acid application (VIA), VIA with magnification (VIAM) and human papillomavirus (HPV)-DNA testing. Colposcopic examination was performed in all and on-site biopsy was taken if any grade I and above lesion was detected on colposcopy (230). On histopathological examination, 105 showed cervical intraepithelial neoplasia II and above lesions. Sensitivity, specificity and predictive values for each test were calculated taking colposcopy and or directed biopsy as the gold standard. Comparisons were made with cytology in order to assess the feasibility of alternative strategies in resource-poor settings. RESULTS: VIA was less sensitive (86.7% versus 91.4%) but more specific (90.7% versus 86.6%) than cytology at low grade squamous intraepithelial lesion (LSIL) threshold but the difference was not statistically significant (P > 0.01). HPV testing improved the sensitivity over cytology (97.1% versus 91.4%) but there was a nonsignificant loss of specificity (84.2% versus 86.6%). Results of VIAM were more or less similar to VIA. CONCLUSION: VIA can be used as a mass screening tool for cervical cancer in resource-poor settings.

Early detection of cervical cancer through acetic acid application--an aided visual inspection.

OBJECTIVE: To assess the sensitivity and specificity of visual inspection of cervix for detection of precancerous and early cancerous lesions of cervix. METHODS: In a Maternal and Child Health Care setting of New Delhi women underwent a detailed pelvic examination, visual inspection of cervix after 5% acetic acid application, cytology (pap smear), detailed colposcopic examination and colposcopic directed biopsy when indicated. RESULTS: Findings of aided visual inspection using 5% acetic acid and of cytology were evaluated among symptomatic 402 women against colposcopic findings and/or histologic reports. Seventy-three mild dysplasias, 50 moderate dysplasias, 45 severe dysplasias/Carcinoma in-situ and 40 early invasive cancerous cases were diagnosed histologically. The sensitivity of cytology (75.3%) was higher compared to that of acetic acid application (52.0%) for mild dysplasias. On the other hand, the sensitivity for detecting moderate dysplasias was 78% for cytology and 81.6% for acetic acid; for severe dysplasias/carcinoma in-situ it was 73.3% for cytology and 86.7% for acetic acid. For invasive cancers sensitivity for acetic acid application and cytology (95% for both modalities) was comparable. The specificity of cytology (99%) was higher compared to that of acetic acid application (94.3%). The false positive rate for cytology was 1.0% as against 5.7% for acetic acid application. The results of acetic acid application also showed a remarkable improvement in the sensitivity of unaided visual inspection for early cancerous lesion which was about 60% for early cancerous lesion and only 12% for mild dysplastic and 20% for moderate and severe dysplastic lesions in our earlier experience. It also reduced the false positive rates from 12% by unaided visual inspection to 5.7% by acetic acid application. Furthermore, cost of detection of one true lesion through acetic acid application (Rs.1689.00) was much lower as compared to the cost involved in cytology detected true lesions (Rs.2227.00). Visual inspection without acetic acid incurred Rs.6608.60 for detection of true lesion. CONCLUSION: Screening for cervical precancerous and cancerous lesions using visual inspection aided by acetic acid may be a suitable low-cost and a feasible alternative modality for control of cervical cancer in a resource poor setting.

Low-cost technology for screening uterine cervical cancer.

We report on an illuminated, low-cost (Rs 1500 (US$ 36)) magnifying device (Magnivisualizer) for detecting precancerous lesions of the uterine cervix. A total of 403 women attending a maternal and child health care clinic who had abnormal vaginal discharge and related symptoms were referred for detailed pelvic examination and visual inspection by means of the device after the application of 5% (v/v) acetic acid. Pap smears were obtained at the same time. The results were compared with those obtained using colposcopy and/or histology. The Magnivisualizer improved the detection rate of early cancerous lesions from 60%, for unaided visual inspection, to 95%. It also permitted detection of 58% of cases of low-grade dysplasia and 83% of cases of high-grade dysplasia; none of these cases were detectable by unaided visual inspection. For low-grade dysplasia the sensitivity of detection by means of the Magnivisualizer was 57.5%, in contrast with 75.3% for cytological examination. However, the two methodologies had similar sensitivities for higher grades of lesions. The specificity of screening with the Magnivisualizer was 94.3%, while that of cytology was 99%. The cost per screening was approximately US$ 0.55 for the Magnivisualizer and US$ 1.10 for cytology.

Biological behavior and etiology of inflammatory cervical smears.

Two hundred and fifty-seven consecutive women attending a major maternal and child health (MCH) center were studied clinically, colposcopically, cytologically, and microbiologically for different gynecologic infections. Out of 257 cases, 207 (80.5%) had inflammatory cervical smears, of which 183 (88.4%) were infected with one or more genital tract infections. Bacterial vaginosis (risk, 22.6-fold), chlamydia (risk, 21.6-fold), and human papillomavirus (HPV) (risk, 13.5-fold) were independently associated with inflammatory smears. In addition, significantly higher proportions of women with inflammatory smears had cervical ectopies (28.5% vs. 10.2%) and bleeding ectopies (30.9% vs. 4.1%) as compared to noninflammatory smears. Women infected with bacterial/parasitic genital infections were given specific treatment. These women were followed up at regular intervals to assess the efficacy of antimicrobial therapy. During follow-up examination, only 26 women (12.6%) showed negative smears. Sixteen women developed squamous intraepithelial lesions (SIL) during follow-up, and 163 women had persistent inflammatory smears. Multivariate analysis revealed that persistent inflammatory smears were associated with herpes simplex virus (HSV) infection, as revealed through detection of IgA antibodies to HSV (risk, 11.5-fold). Progression of SIL was associated with HPV infection (risk, 17.6 fold). Thus, inflammatory smears are associated with different types of infection, most of which do not respond to antimicrobial therapy.

Colposcopy as a tool for detection of human papillomavirus infection of uterine cervix in the setting of high prevalence of gynaecologic infections.

A cross sectional study involving 257 women from the Maternal and Child Health Centre (MCH) in Delhi was initiated for screening clinically, cytologically, colposcopically, and microbiologically for several gynaecologic infections. Eighty percent of the women had one or more gynaecologic infections and 31.1% had three or more infections. Cytology revealed changes suggestive of condyloma in 3 (1.2%) women only, while colposcopic examination suggested HPV changes in 117 (45.5%) women. A very high proportion of colposcopically detected lesions (78.6%) had evidence of HPV related changes in histology. The specificity of these lesions were further confirmed by Pan HPV DNA in-situ hybridisation, when 84% of the colposcopy is a valuable tool for detecting subclinical HPV lesions in a setting with high prevalence of gynaecologic infections.

Mutagenicity of endocervical mucus associated with genital tract infections.

Mutagenic metabolites produced due to chronic infection of cervical epithelium are suspected to be a plausible risk factor in cervical carcinogenesis. One hundred twenty-four symptomatic women attending a maternal and child health (MCH) clinic were studied clinically, cytologically, microbiologically, and biochemically for genital tract infections and for the presence of mutagens in the endocervical secretions using Ames' test. Human papillomavirus (HPV) was the leading infection (53.3%), followed by chlamydial infection (25.8%) and seropositivity for herpes simplex virus (25.1%). Mutagenic products in the endocervical secretions was detected in 23 women (18.5%). The univariate and multivariate (adjusted for other genital infections, age, and parity) analysis showed that the mutagenic mucus was associated with only chlamydial infection of endocervical region (OR = 3.7; 95% CI = 1.7,7.3). This shows that chlamydia is associated with mutagenicity of endocervical mucus.

Significance of inflammatory cervical smears.

Two hundred and fifty-seven women attending a Maternal and Child Health Centre (MCH) were examined for different colposcopic and histological patterns associated with cervical inflammation as detected by cytology and for their association with different gynaecological infections. The cytodiagnosis revealed inflammation in 207 women (80.5%) and non-inflammation in 49 (19.5%); one smear was inadequate for evaluation. Fifty-six per cent of the women with inflammation and 20% with non-inflammation had an atypical transformation zone (ATZ), the risk of ATZ being 4.9-fold higher in those with inflammation. Biopsies from 128 women with abnormal colposcopy revealed morphological changes suggestive of human papillomavirus (HPV) in 89 (69.5%) and dysplasia of varying grades in 8 (6.3%). Seventy per cent of histologically diagnosed HPV lesions stained immunohistochemically, whereas 84% reacted with a biotinylated Pan-HPV probe by DNA in situ hybridization (DISH). In addition to HPV, chlamydia (OR 15.6, 95% CI 2.2, 311.6), T. vaginalis (OR 18.4), bacterial vaginosis (OR 24.7, 95% CI 3.5, 492) and herpes simplex virus (OR 4.9, 95% CI 1.4, 20.9) were significantly associated with inflammatory smears. Of 11 dysplasias detected by colposcopy and confirmed by biopsy, 8 (72.7%) had inflammatory cytology in the initial Pap smears. Thus a large proportion of women with inflammatory smears had multiple gynaecological infections and may be at increased risk of developing preneoplastic or neoplastic changes. Furthermore, they risk transmitting the infections to their partners.

Clinical presentation of gynecologic infections among Indian women.

OBJECTIVE: To study the clinical presentation of different gynecologic infections among Indian women. METHODS: This was a cross-sectional study of 257 women that included clinical, cytologic, colposcopic, and microbiologic screening for various gynecologic infections. RESULTS: Human papillomavirus (HPV) was the leading infection, affecting 127 (49.4%) women; however, overt warts were only seen in seven (2.7%) patients. Women infected with HPV had a 60.3-fold higher risk of developing a bleeding ectopia compared to those with other infections; women with an unhealthy cervix and cervical ectopias also had an increased risk of HPV infection (7.6- and 2.8-fold, respectively). Bacterial vaginosis, detected in 33.5% of the women studied, had an increased risk of bleeding ectopia (9.3-fold), cervical ectopia (3.1-fold), cervicitis (2.9-fold), vaginitis (6.9-fold), and cervical hypertrophy (2.1-fold). Chlamydial infection, detected in 23.3% of the patient population, was associated with an eightfold increase in the risk of an unhealthy cervix and a fourfold increase in risk of a hypertrophied cervix. Immunoglobulin-A antibodies to the herpes simplex virus were detected in 53 (20.6%) women. More than half (55.2%) of the women had two or more infections, and the mean delay of seeking medical treatment was 7-13 months. CONCLUSION: The specific finding of bleeding cervices was associated with HPV and bacterial vaginosis, hypertrophied cervices with chlamydia and bacterial vaginosis, and unhealthy cervices with chlamydia and HPV infections.

Association between reproductive tract infections and cervical inflammatory epithelial changes.

OBJECTIVE: To determine the association, if any, between different reproductive tract infections and cervical inflammatory epithelial changes. STUDY DESIGN: Clinical, colposcopic, cytologic, and microbiologic screening for reproductive tract infections was conducted. Subjects were screened for Chlamydia trachomatis, Neisseria. gonorrhoeae, Trichomonas vaginalis, bacterial vaginosis, yeast vaginitis, human papillomavirus, and serology for herpes simplex viruses, syphilis, and human immunodeficiency virus-1 and -2. The subjects were 257 women who visited a maternal and child health center between January 1992 and December 1993. RESULTS: Cervical cytology revealed inflammatory epithelial changes in 207 women (80.5%), the highest proportion in the published series. Inflammatory epithelial changes were significantly associated with the number of reproductive tract infections. Risk increased up to 72.6 fold when women were infected with two or more agents. The positive predictive value of inflammatory epithelial changes for any infection was 88.4%, whereas the negative predictive value for any infection was 63.4%. In univariate analysis, the cervical inflammatory epithelial changes were significantly associated with infections such as chlamydia (28.0%), human papillomavirus (56.5%), Trichomonas vaginalis (18.3%), bacterial vaginosis (38.2%), and herpes simplex virus as evidenced by the presence of immunoglobulin M antibodies (28%). Multivariate analysis, however, revealed an independent association of inflammatory epithelial changes with chlamydia (odds ratio, 21.3; 95% confidence interval, 2.6, 181.3), human papillomavirus (odds ratio, 13.5; 95% confidence interval, 4.5, 39.6), and bacterial vaginosis (odds ratio, 22.6; 95% confidence interval, 2.9, 181) only. CONCLUSION: Cervical inflammatory epithelial changes are significantly associated with reproductive tract infection. Cervical inflammatory epithelial changes predict correctly to an extent of 81% infections with chlamydia, bacterial vaginosis, and human papillomavirus infection. A negative smear, on the other hand, rules out these infections to an extent of 83%.

Circulating immune profile in patients with pre-cancer and cancer of the cervix: a cross sectional study among Indian women.

To fully understand immune competence in subjects with precancerous and cancerous lesions of the uterine cervix, various markers of T and B cells in peripheral blood were evaluated. The cellular measures were the absolute numbers of CD2, CD3, CD22, CD25, CD4, CD8 and CD4/CD8 T cell ratio and serologic measures were the levels of cellular products ie interleukin-2 (IL-2) and soluble interleukin-2 receptors (SIL2-R). Eighty-five subjects were analysed, and diagnosed histologically as CIN (n = 33) and invasive cancer (n = 22). Thirty women with normal Pap test results served as controls. Results on cellular markers showed a significant decrease in peripheral blood lymphocyte counts in patients with higher grades of cervical lesions as compared to controls (CIN III P < 0.05 and invasive cancer P < 0.001). Similar trends were observed for CD22 cell counts (P < 0.001). A consistent decrease was observed for CD2 positive T lymphocytes in patients with higher grade cervical lesions only. In addition, a decrease in CD4-T cells with a relative increase in CD8 and CD25 cell count was observed in these groups of patients, leading to a considerable reduction in CD4/CD8 T-cell ratio. Results obtained on soluble cellular products revealed that serum levels of IL-2 and SIL2-R in patient groups were significantly higher than controls (P < 0.001). Preliminary findings indicated a host-vested cellular impairment, evidenced by immune deficit and immune-stimulation, in women with pre-cancer and cancerous lesions of the cervix.

HSV-IgA serum antibodies in cervical intraepithelial neoplasia and invasive cancer patients, and in their spouses: a case control study.

Class-specific IgG and IgA antibodies to HSV were assayed in women with CIN (76), invasive cancer (52) (histological diagnosis) and age-matched controls (119), employing HSV-2-infected HEp-2 cells as antigen during IFA assay. We observed an elevated geometric mean titre (GMT) of serum antibody (IgG five-to eight-fold and IgA four-to five-fold) for the entire spectrum of cervical lesions, as compared to controls. The odds of finding HSV-IgA antibodies were highest with CIN III (OR = 22.0), followed by invasive carcinoma, and CIN I & II (OR = 9.5 and 5.2), respectively. Furthermore, the investigations with respect to married couples (husbands and wives) who volunteered to participate in this study (33 cases and 47 control group) also indicated relatively high antibody titres and increased frequency of HSV sero positivity amongst husbands of cases as compared to their wives, as well as the control group males and females. The contribution of HSV infection in women and/or their husbands to the risk of developing abnormal cervical lesions was analysed after adjusting for the same in respective counterparts. It was observed that the risk was increased 14-fold with HSV-IgA positivity of women, and that HSV-IgA positivity of husbands (male partners) further increased the risk 16-fold. This preliminary observation shows the importance of serum HSV-IgA antibodies as a risk indicator in cervical precancer and cancer lesions in women without a history of recent genital herpes lesions. The serum HSV-IgA may also be taken as an indicator of "high risk" males.

Peripheral blood lymphocyte subpopulations in Indian women with cervical intraepithelial neoplasia and invasive cancer--an immunocytochemical study using monoclonal antibodies.

The peripheral blood lymphocyte subpopulations in Indian women with various grades of cervical intraepithelial neoplasia [33], invasive cancer [22] and matched controls [30] were studied using specific monoclonal antibodies and biotin avidin immunofluorescence technique. Our results showed a significant fall in peripheral T (CD2) B(CD22) cells in patients with CIN III and invasive cervical cancer lesions compared to controls (P less than 0.001). Similarly the quantum of T helper cells (CD4) decreased according to the severity of cervical lesions (P less than 0.01 - P less than 0.001), whereas the T suppressor cells (CD8) depicted an increasing trend in patients with higher grade of cervical lesions (CIN III and invasive cancer, P less than 0.001) as compared to controls. Further, the CD4/CD8 ratio showed a significant downfall with the severity of cervical lesions (P less than 0.01 - P less than 0.001), indicating a perturbance in the homeostasis of host cellular immunity.

Detection of mutagenicity in cervico-vaginal secretions--a plausible risk factor for cervical cancer.

The cervico-vaginal secretions from 51 women with various grades of dysplastic lesions of uterine cervix were assessed for mutagenic potential by Ames test using histidine deficient mutant strain of Salmonella typhimurium TA-98: with S-9 mix. Twenty three per cent of samples from women with cervical dysplasia were found significantly positive (P less than 0.001) for mutagenic activity compared to 3% positive from control. The frequency of mutagenic secretions detected were almost uniform, irrespective of the severity of cervical lesions. None of cervico-vaginal secretions, positive for mutagenicity could revert the tester strain when tested in absence of S-9 mix (liver microsomal enzymes). This indicates that mutagens in cervico-vaginal secretions are effective only when activated enzymatically.