Impact of preterm birth on the cellular characteristics of neonatal buccal cells.

OBJECTIVE: To demonstrate the impact of preterm birth on the cytological, cytomorphometrical, and nuclear parameters of neonatal buccal smears. METHODS: This study consisted of Early Preterm Neonates (EPN; ≤34th gestational week [gw]; n = 36), Late Preterm Neonates (LPN; 34th to <37th gw; n = 46), and Term Neonates (control; ≥37th gw; n = 56). Cytological evaluation and buccal cytome assay were performed using Papanicolaou and Feulgen methods, respectively. RESULTS: Cytological evaluation demonstrated that smear background was cleaner (P < .05) and there were less macrophages in the control group (P < .001). Cyto-morphometric analysis showed that the measurements of nuclear diameter, nuclear area, and nucleus-to-cytoplasm ratio were higher in the preterm (EPN and LPN) versus the control groups (P = .016, P < .001, and P < .001, respectively). We also demonstrated that staining intensity of the nucleus and cytoplasm were less intense in the EPN and LPN groups (P < .001). There was no statistically significant difference between the EPN and LPN groups for any parameters (P > .05). Buccal cytome assay showed that nuclear buds were more prevalent in term newborns compared to preterm neonates (P < .001). CONCLUSIONS: Morphological and cytological properties of neonatal buccal cells are influenced by preterm birth status, and buccal smears may be used as a tool to detect biological markers of neonatal health problems.

Determination of Barr bodies in Transgender Patients in India - A comparative study.

Sex determination in forensic medicine is considered one of the first and foremost steps in personal identification. The need for identifying the exact sex of the individual arises when deciding whether a person can exercise certain civil rights reserved for one particular sex, for competing in sex-specific athletic and sports events, legitimacy, divorce, paternity disputes and also to some criminal offenses. Nuclear sexing by Barr body examination can be done using buccal smears to establish the sex of the individual when routine methods fail to disclose the exact gender of the individual. Aim: To determine and compare the Barr bodies present in exfoliated buccal epithelial cells in males, females and transgender populations using light and fluorescence microscopy. Materials and Methods: A total of 90 patients were recruited for the study. Group I consisted of 30 female patients. Group II consisted of 30 male patients and group III consisted of 30 transgender patients. The buccal mucosa was then scraped using a wooden spatula and the cells obtained were fixed in 95% ethanol. Two smears per individual were made and stained. One smear was stained with papanicolaou (PAP) stain and the other with Acridine orange and viewed under light microscopy and fluorescent microscopy, respectively. Results: When PAP stained slides were examined, the percentage of Barr-bodies in females ranged from 3% to 5% and in males it was 0% and in transgenders, it ranged from 0% to 5%. In Acridine orange stained smears, the percentage of Barr bodies in females ranged from 1% to 3% and in males it was 0% and in transgenders, it was 0%. Kruskal-Wallis test to study the relation of Barr body percentage in females, males and transgender subjects demonstrated significant differences between the groups (P < 0.001). Wilcoxon signed rank test was done for pairwise comparison, which showed that the distribution of percentage of positive cells in females are statistically significant from males and transgenders (P < 0. 001). Conclusion: Nuclear sexing using Barr bodies offers a simple yet effective method for determining the sex of transgender patients which could help them in understanding their gender identity better and diagnose any underlying chromosomal aberration.

Comparative Analysis of Cytomorphology in Adolescent and Geriatric Patients: A Cross-Sectional Study.

Background Exfoliation of the surface cells of the normal epithelium occurs as a result of physiological turnover. Epithelial cells are constantly renewed, and they are exfoliated or shed as they migrate from the basal layer to the uppermost layer of the epithelium. Oral exfoliative cytology involves the collection and microscopic evaluation of these shed cells or scraped epithelial cells, quantitatively and qualitatively. The objective of the present study was to analyze and compare the cytomorphometric features like cellular area, nuclear area, and nuclear-cytoplasmic ratio from buccal mucosal smears of adolescent and geriatric patients. This study highlights the changes in cell morphology in different age groups (adolescent and pediatric), which could be attributed to hormonal, habitual, and aging factors.  Material and methods Buccal smear sample was collected from a total of 60 individuals belonging to the age groups of 11-19 years and above 60 years. The smears were stained with H&E and PAP (Papanicolaou) stain. Photomicrographs were taken in 40x, and measurements were calculated using ImageJ software (National Institutes of Health, Bethesda, Maryland, United States). Cellular size, nuclear size, and nucleo-cytoplasmic ratio were analyzed and compared between the two age groups using independent t-tests using IBM SPSS Statistics for Windows, Version 22.0 (Released 2013; IBM Corp., Armonk, New York, United States). Results A significant difference was observed in the cellular area and nuclear area between the two age groups with a p-value of 0.00. No significant findings were present in the nucleo-cytoplasmic area of the two age groups. Conclusion Cytomorphometric analysis has shown that there were variations in the cytoplasmic and nuclear areas among different age groups.

Cytomorphometric Analysis of Buccal Exfoliated Cells in Geriatric and Pediatric Age Groups - A Cross-Sectional Study.

Background We have to establish variations in cellular dimensions in buccal smears with respect to age. It can be used as a reference standard while dealing with age-related pathological abnormalities. Aim The study aims to compare the nuclear area (NA), cellular area (CA), and nucleus-to-cytoplasm ratio (N:C) of pediatric and geriatric age groups in smears obtained from clinically normal buccal mucosa. Materials and methods Buccal smears were collected from 60 subjects with age groups of <10 and >60 years. Cytological smears were prepared and fixed using alcohol. H&E and Papanicolaou's staining was performed as per manufacturer instructions. Cytomorphometric analysis was carried out using Image J software v.1.52 for CA, NA, and N:C. Statistical analysis using Student's t-test was performed using SPSS version 23.0 (IBM Inc, Armonk, New York).  Results A significant difference (p<0.001) in NA and CA between the pediatric and geriatric age groups was noted. There was no significant difference in N:C among the study groups. Conclusion The present study provides baseline data of two different age groups that can be used for comparison of abnormal cells in suspicious clinical lesions.

Multi-tissue cytogenetic analysis for the diagnosis of mosaic Down syndrome: A case report.

Less than one percent of individuals with Down syndrome exhibit mosaicism, a biological phenomenon that describes an individual who has two or more genetically distinct cell lines. The percentage of mosaicism in different tissues can impact the presence of clinical findings and hinder cytogenetic diagnosis. We report a case of mosaicism for trisomy 21 diagnosed after multi-tissue cytogenetic analysis of peripheral blood and buccal mucosa, associated with significant intellectual disability, dysmorphic facial features, congenital heart defects, macropenis, and imperforate anus.

Genotoxicity in Oral Mucosal Epithelial Cells of Petrol Station Attendants: A Micronucleus Study.

INTRODUCTION: Occupational exposure to petrol derivatives possesses an increased risk of various cancers including that of the oral mucosa. Scientific studies have shown the correlation of micronuclei assay (MN) with the cytogenotoxic changes in petrol station attendants. However, very few have reported the use of MN assay as a promising tool for assessing the impact of smoking in these workers. AIM: To explore the cytogenotoxic damage in exfoliated buccal cells obtained from petrol station attendants and control subjects using the MN assay along with additional effects due to smoking. MATERIALS AND METHODS: The study comprised 60 males who were divided into Group I-IV with each having 15 subjects. These subjects were categorized as exposed smokers, exposed nonsmokers, unexposed smoker group, and unexposed nonsmokers. The MN and additional nuclear abnormalities (karyorrhexis [KH], binucleation [BN], pyknosis [P], and karyolysis [KL]) were calculated in PAP-stained slides. RESULTS: Statistically higher mean frequencies of overall nuclear anomalies were observed in petrol pump workers in comparison with the control group. Petrol pump smokers carry the highest nuclear anomalies followed by non-exposed smokers than exposed non-smokers and the count was the least among unexposed non-smoker workers. DISCUSSION AND CONCLUSION: The present study indicated that the petrol pump workers are under higher cytogenotoxic damage. Also, smoking added to the frequency of damage. Thus, MN and other nuclear anomalies are in-vitro reliable biomarker assays available and should be routinely employed as a screening tool in their periodic medical evaluation.

Reactivation of inactive X chromosome in buccal smear of carcinoma of breast.

Buccal mucosal smears of 100 female patients of carcinoma of breast were compared with 100 controls matched accordingly. The frequency of Barr bodies was significantly lower in carcinoma of breast patients (menstruating and menopausal women) P < 0.001 when compared with controls indicating reactivation of the inactive X chromosome.

Sex determination efficacy of Papanicolaou and acriflavine Schiff stains in buccal smears.

BACKGROUND: Barr body is formed from random inactivation and condensation of one of the two female chromosomes in virtually all the somatic cells of female mammals. Buccal smears have been reported to be potential sources of Barr bodies. AIM: This study was done to assess the efficacy of acriflavine (AF) Schiff and Papanicolaou (PAP) stains in sex determination by identifying Barr bodies in buccal smears of both sexes. MATERIALS AND METHODS: Two samples of buccal smears, collected from thirty males and thirty females in the age group of 16-60 years were used to demonstrate Barr bodies using AF Schiff and PAP stains, respectively. Hundred cells were examined for Barr body positive nucleus, and its mean percentage was calculated and statistically analyzed. RESULTS: In females, AF Schiff stained positive cells ranged from 16% to 53% and PAP stained positive cells ranged from 9% to 38%. In males, 0-9% AF positive Barr bodies and 0-5% PAP stained Barr bodies were identified. CONCLUSION: Sex determination using buccal smear is a simple and reliable method. AF Schiff stain is better both qualitatively and quantitatively when compared to PAP stain, thus aids in more accurate sex determination.

Rare case of Killian-Pallister syndrome associated with idiopathic short stature detected with fluorescent in situ hybridization on buccal smear.

BACKGROUND: Killian-Pallister syndrome (KPS) is a rare form of chromosomal mosaicism and is defined by the existence of an extra chromosome 12 in some cell lines in one individual. The degree of mosaicism varies among tissues and dictates the clinical presentation of the syndrome. The clinical features of Killian-Pallister syndrome include mental retardation, typical facial dysmorphism and pigmentation defects. CASE PRESENTATION: We present a rare case of Killian-Pallister syndrome with severe form of the disease associated with isolated growth hormone deficiency and low-rate mosaicism on buccal smear. The absence of a marker chromosome 12p in lymphocyte cultures and the low degree of mosaicism lead to frequent misdiagnosis of this condition. CONCLUSIONS: The selection of tissue sampling is crucial in establishing the diagnosis of Killian-Pallister syndrome. Fluorescent in situ hybridisation on buccal smear remains the golden standard as a screening method if a suspicion of the syndrome exists.

Cytogenomic delineation and clinical follow-up of 10 Brazilian patients with Pallister-Killian syndrome.

BACKGROUND: Pallister-Killian syndrome (PKS) is a sporadic genetic disorder caused by the presence of a tissue-specific mosaicism for isochromosome 12p - i(12) (p10) and is characterized by facial dysmorphism including coarse facies, upslanting palpebral fissures, bitemporal alopecia, pigmentary skin anomalies, developmental delay, hypotonia and seizures. Although typical clinical features of PKS commonly exist, clinicians often do not raise the possibility of this diagnosis. RESULTS: We reviewed the medical records of 10 patients with confirmed PKS followed in our service (since 1990 to 2015). Age at diagnosis varied from prenatal to 3 years and clinical features were consistent with those described in the literature. In all patients, peripheral blood karyotypes were normal and cytogenomic study was performed in order to confirm the diagnosis. Three of these patients had PKS diagnosis confirmed by buccal smear MLPA. CONCLUSION: An early conclusion from our results demonstrated that MLPA on buccal smears is a good and non-invasive method to detect extra copies of 12p and should be considered as the first exam, before a skin biopsy for a fibroblast karyotype is performed.

Oral Cytology for HIV: A New Diagnostic Tool?

AIM: To asses morphological and morphometrical alterations of oral squamous epithelial cells in HIV infected individuals, and determine its diagnostic significance for HIV infection. MATERIALS AND METHODS: Informed Consent was obtained from all study subjects. 40 adult HIV infected patients (experimental group) and 40 adult non-infected volunteers (control group) participated. Age, occupation, and relevant medical history were recorded. The following laboratory tests reports were recorded: complete blood cell counts, blood glucose levels, serum iron and erythrocyte sedimentation rate. Oral smears were collected from normal appearing tongue and buccal mucosa of the individuals by exfoliative cytology. The cells were morphologically analysed and the nuclear area (NA), the cytoplasmic area (CA) and the nucleus-to-cytoplasm area ratio (NA/CA) were calculated. Cell yield, cell cohesion, presence/absence of inflammatory cells and candida were observed. RESULTS: The cytological smears of HIV patients showed abundant cell yield and the epithelial cells were found to be in close cohesive clusters and both findings were found to be statistically significant. Nuclear cytoplasmic ratio was seen to be increased in 93.8% of HIV positive patientsand Mild to moderate pleomorphism was observed in 17.9% of HIV positive patients. HIV positive patients had a diminished inflammatory response and this was found to be statistically significant. CONCLUSION: Statistically significant deviations from normal oral epithelium were found in the study conducted. With further research, oral exfoliative cytology may form a new, painless, inexpensive diagnostic method for HIV infections.