Micronucleus in oral exfoliated cells and associated factors among young adults in the far south of Brazil

There is an increasing interest in the application of micronucleus (MN) assays in the clinical setting to investing the impact of environmental, genetic, life style and socioeconomic conditions on DNA. However, the evaluation of mutagenic damage among young population had not been studied and early intervention in this population can prevent several health comorbidities. Thus, the present study aimed in investigate the MN presence in young adults and its associated factors. This is a cross-sectional study, composed by 155 young adults (between 18 to < 30-year-old) of both sexes. The socioeconomic and life style data was collected through a semi-structured questionnaire. Oral exfoliated cells were collected to evaluate the frequency of MN cells. Socioeconomic conditions and life style factors were not associated with the presence of MN. Twenty percent of the participants reported some chronic noncommunicable disease and almost twelve percent reports continued use of medication, both variables present a risk factor to prevalence of MN. Although mutagenesis is a process generally more related to older individuals, this study highlights the importance of monitored DNA damage between young adults and promote healthy habits to prevent the development and worse prognosis of chronic non-transmissible diseases and the consequent use of drugs.

Investigations concerning the impact of consumption of hot beverages on acute cytotoxic and genotoxic effects in oral mucosa cells

Consumption of very hot beverages and foods increases the incidence of oral and esophageal cancer but the mechanisms are not known and the critical temperature is not well defined. We realized a study with exfoliated cells from the oral cavity of individuals (n = 73) that live in an area in Iran which has the highest incidence of EC worldwide. Consumption of beverages at very high temperatures is a characteristic feature of this population. We analyzed biomarkers which are (i) indicative for genetic instability (micronuclei that are formed as a consequence of chromosomal damage, nuclear buds which are a consequence of gene amplifications and binucleated cells which reflect mitotic disturbances), (ii) markers that reflect cytotoxic effects (condensed chromatin, karyorrhectic, karyolitic and pyknotic cells), (iii) furthermore, we determined the number of basal cells which is indicative for the regenerative capacity of the buccal mucosa. The impact of the drinking temperature on the frequencies of these parameters was monitored with thermometers. We found no evidence for induction of genetic damage but an increase of the cytotoxic effects with the temperature was evident. This effect was paralleled by an increase of the cell division rate of the mucosa which was observed when the temperature exceeded 60 °C. Our findings indicate that cancer in the upper digestive tract in drinkers of very hot beverages is not caused by damage of the genetic material but by an increase of the cell division rate as a consequence of cytotoxic effects which take place at temperatures over 60 °C. It is known from earlier experiments with rodents that increased cell divisions lead to tumor promotion in the esophagus. Our findings provide a mechanistic explanation and indicate that increased cancer risks can be expected when the drinking temperature of beverages exceeds 60 °C.

The Value of MicroRNA-375 Detection for Triaging Primary Human Papillomavirus Positive Women: A Cross-Sectional Study in a General Population

PurposeThis study aims to validate the value of microRNA (miRNA) detection for triaging human papillomavirus (HPV)-positive women in the general population.Patients and MethodsmiR-375 detection in cervical exfoliated cells has been demonstrated to have the superior value to cytology in triaging primary HPV-positive women in the hospital population. In this study, residual samples of cervical exfoliated cells from 10,951 women in a general population were used to detect miRNA. The performance efficiency of miRNA detection in identifying high-grade cervical intraepithelial neoplasia (CIN) was evaluated. Pearson chi-square test and McNemar pairing test were used to compare miRNA detection and cytology.ResultsIn valid 9,972 women aged 25–65, miR-375 expression showed a downward trend along with an increase in cervical lesion severity. The expression level of miR-375 ≤1.0 × 10-3 was identified as positive. In the HPV-positive and 12 HPV genotypes other than 16/18 (HR12)-positive women, miR-375 detection showed equivalent sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) to that of cytology (≥ASC-US) and higher or similar sensitivity and NPV but lower specificity and PPV than that of cytology (≥ASC-H) in identifying CIN3+ and CIN2+. In HPV 16-positive women, miR-375 positivity had higher sensitivity and NPV but lower specificity and PPV than that of cytology (≥ASC-H and HSIL) in identifying CIN3+ and CIN2+. The immediate CIN3+ risk of miR-375 positivity was 19.8% (61/308) in HPV-positive, 10.8% (22/204) in HR12-positive, and 43.5% (37/85) in HPV16-positive women, respectively.ConclusionThe detection of miR-375 in cervical exfoliated cells may be an optional method for triaging primary HPV-positive women in population-based cervical cancer screening.

Low-Coverage Whole Genome Sequencing Diagnoses Endometrial Carcinoma from Tampon DNA of Uterine Bleeding Patients

Background Endometrial carcinoma (EC) is a disease predominantly affecting postmenopausal women. It accounts for about 5% of abnormal uterine bleeding. It is still challenging to diagnose cancers from uterine bleeding patients. Previously, chromosome aberrations were found to be frequent in EC. Here we employed a low coverage whole genome sequencing technology to investigate chromosome aberrations in tampon-collected DNA of patients with suspicious EC. Methods Thirty ECs and 14 benign cases with abnormal bleeding are prospectively recruited. Tampons were used to collect exfoliated cells and DNA extracted from these exfoliated cells was analyzed by a customized bioinformatics workflow, uterine exfoliated cell chromosomal aneuploidy detector (UterCAD). Results As our data shown, frequent chromosome copy number variations (CNV) were found in EC patients as compared to non-tumor controls, especially the chromosome 8q gain and 10q gain. Using UterCAD, CNVs were detected in tampon-derived DNA from 83.3% (25/30) EC, which were 80.8% (21/26) EECs and 100% (4/4) USCs. In EEC group, CNVs were found in 81.3% (13/16), 85.7% (6/7), and 66.7% (2/3) patients of stage IA, IB, and II/III, respectively. Moreover, all the 4 USC patients presented significant CNVs. Conclusions UterCAD could be a highly specific, robust uterine cancer diagnosis method, with an especially high sensitivity for the more aggressive subtype - serous carcinoma. It may be used as a non-invasive approach for diagnosis and active surveillance in endometrial cancer prior to the use of biopsy, thereby largely reducing the treatment burden on patients.

AN INTERVENTIONAL STUDY TO ASSESS THE CYTOLOGICAL AND COLPOSCOPIC CHANGES AFTER INTRAUTERINE CONTRACEPTIVE DEVICES INSERTION IN THE DEPARTMENT OF OBSTETRICS GYNAECOLOGY, SMS MEDICAL COLLEGE, JAIPUR

Background: Cytology examines exfoliated cells, while colposcopy examines the changes that occur in the vascular structure of the cervix5. Patients with an abnormal pap smear result or patients with a suspicious-looking cervix even if they had a negative pap smear, should be evaluated by colposcopy and colposcopy directed biopsy. Methods: Interventional Prospective study conducted at Department of Obstetrics and Gynaecology, SMS Medical College & associated Hospitals, Jaipur. Results: In this study, among 15 IUCD users, 5 (33.33%) had bacterial vaginosis, 2 (13.33%) had trichomonas vaginalis and 1 (6.66%) were found candidal infection, 1 (6.66%) polymicrobial infection. Conclusion: The present study indicates that there is definite changes in the cervical architecture in IUCD users and the change is also dependent on the duration of use. A simple technique like cytology and colposcopy is highly useful in diagnosing these changes and combating them by early interference whenever necessary. The present study does not reveal any increased risk of cervicitis and dysplasia in IUCD users. We further recommend that a long term follow-up is required to rule out the possibility of any increased risk of dysplasia or carcinoma in-situ. Keywords: IUCD, Colposcopy, Dysplasia.

Rationale and design of the Prevent Anal Cancer Self-Swab Study: a protocol for a randomised clinical trial of home-based self-collection of cells for anal cancer screening

IntroductionSquamous cell carcinoma of the anus is a common cancer among sexual minority men, especially HIV-positive sexual minority men; however, there is no evidenced-based national screening protocol for detection of anal precancers. Our objective is to determine compliance with annual anal canal self-sampling or clinician-sampling for human papillomavirus (HPV) DNA.Methods and analysisThis is a prospective, randomised, two-arm clinical study to evaluate compliance with annual home-based versus clinic-based HPV DNA screening of anal canal exfoliated cells. The setting is primary care community-based clinics. Recruitment is ongoing for 400 HIV-positive and HIV-negative sexual minority men and transgender persons, aged >25 years, English or Spanish speaking, no current use of anticoagulants other than nonsteroidal anti-inflammatory drugs and no prior diagnosis of anal cancer. Participants are randomised to either receive a swab in the mail for home-based collection of an anal canal specimen at 0 and 12 months (arm 1) or attend a clinic for clinician collection of an anal canal specimen at 0 and 12 months (arm 2). Persons will receive clinic-based Digital Anal Rectal Examinations and high-resolution anoscopy-directed biopsy to assess precancerous lesions, stratified by study arm. Anal exfoliated cells collected in the study are assessed for high-risk HPV persistence and host/viral methylation. The primary analysis will use the intention-to-treat principle to compare the proportion of those who comply with 0-month and 12-month sampling in the home-based and clinic-based arms. The a priori hypothesis is that a majority of persons will comply with annual screening with increased compliance among persons in the home-based arm versus clinic-based arm.Ethics and disseminationThe study has been approved by the Medical College of Wisconsin Human Protections Committee. Results will be disseminated to communities where recruitment occurred and through peer-reviewed literature and conferences.Trial registration numberNCT03489707.

HISTOPATHOLOGICAL GRADING OF ORAL SQUAMOUS CELL CARCINOMA BY MICRONUCLEI ASSAY, A POTENTIAL BIOMARKER IN ORAL EXFOLIATED CELLS

Background: Micronucleus is a microscopically visible round or oval cytoplasmic chromatin mass in the extra nuclear vicinity, originated from aberrant mitosis, which consists of eccentric chromosomes that have failed to reach spindle poles during mitosis and are used as biomarkers for assessment of DNA damage. Micronuclei in exfoliated oral epithelial cells have been shown in some studies to correlate with severity of this genotoxic damage. This severity can be measured in terms of grading of the lesions. Aim: To correlate frequency of micronuclei (MN) in oral exfoliated cells in clinically diagnosed cases of oral squamous cell carcinoma (OSCC) followed by a histopathological grading. Materials and Methods : The study subjects consisted of clinically diagnosed cases of OSCC. Healthy subjects without any tobacco consumption habits formed the control group.The cytosmears from both groups were stained with rapid Papanicolaou stain. MN were identified according to the criteria given by Tolbert et al. Results: The frequency of MN was three to four times higher in patients with OSCC as compared to patients in the control group and the difference was found to be highly significant. In 75% cases, the cytological grade as determined by the frequency of micronuclei correlated with the histopathological grade and this observation was statistically significant. Conclusions : The MN is potentially an excellent candidate to serve as such a biomarker for prediction of the grade of OSCC.