Automated Analysis of Nuclear Parameters in Oral Exfoliative Cytology Using Machine Learning.

BACKGROUND: As oral cancer remains a major worldwide health concern, sophisticated diagnostic tools are needed to aid in early diagnosis. Non-invasive methods like exfoliative cytology, albeit with the help of artificial intelligence (AI), have drawn additional interest. AIM: The study aimed to harness the power of machine learning algorithms for the automated analysis of nuclear parameters in oral exfoliative cytology. Further, the analysis of two different AI systems, namely convoluted neural networks (CNN) and support vector machine (SVM), were compared for accuracy. METHODS: A comparative diagnostic study was performed in two groups of patients (n=60). The control group without evidence of lesions (n=30) and the other group with clinically suspicious oral malignancy (n=30) were evaluated. All patients underwent cytological smears using an exfoliative cytology brush, followed by routine Hematoxylin and Eosin staining. Image preprocessing, data splitting, machine learning, model development, feature extraction, and model evaluation were done. An independent t-test was run on each nuclear characteristic, and Pearson's correlation coefficient test was performed with Statistical Package for the Social Sciences (SPSS) software (IBM SPSS Statistics for Windows, Version 28.0. IBM Corp, Armonk, NY, USA). RESULTS:  The study found substantial variations between the study and control groups in nuclear size (p<0.05), nuclear shape (p<0.01), and chromatin distribution (p<0.001). The Pearson correlation coefficient of SVM was 0.6472, and CNN was 0.7790, showing that SVM had more accuracy. CONCLUSION: The availability of multidimensional datasets, combined with breakthroughs in high-performance computers and new deep-learning architectures, has resulted in an explosion of AI use in numerous areas of oncology research. The discerned diagnostic accuracy exhibited by the SVM and CNN models suggests prospective improvements in early detection rates, potentially improving patient outcomes and enhancing healthcare practices.

Evaluation of nuclear morphometry in exfoliative cytology of buccal mucosa in patients with high risk of oral cancer.

BACKGROUND: Oral cancer poses a significant global health burden, with India having the highest prevalence. Effective detection is crucial in effective prevention. This study aimed to evaluate nuclear morphometric parameters (NMPs) in buccal mucosa cells of smokers, correlate NMPs with dysplasia, establish cut off values for grading dysplasia, and investigate the relationship between NMPs and smoking. METHODS: After obtaining ethical approval and informed consent, patients were recruited from the outpatient department of our institution. A target sample size of 250 was calculated. The data included smoking exposure quantified in pack-years, nuclear morphometric analysis (NMA) of buccal mucosa cells obtained through oral cytology using Image J, and the severity of dysplasia of the slides assessed by pathologists. Statistical analysis assessed the impact of dysplasia and the association between nuclear characteristics and smoking exposure. Receiver operating characteristic (ROC) plots determined the potential of these parameters to distinguish dysplasia levels. RESULTS: Significant differences in NMPs were observed among different smoking groups. Dysplasia severity had a significant correlation with NMPs, and strong correlations were found between NMPs and lifetime smoking exposure. ROC analysis established cut off values for NMPs with good sensitivity and specificity for classifying dysplasia severity. CONCLUSIONS: This study highlights the potential of NMA as a tool for oral cancer screening. NMPs can distinguish dysplasia severity and correlate with tobacco (smoking). The efficiency of NMA in a non-invasive oral cytology offers promise for patient-centered screening Additionally, the findings suggest future applications in telepathology and the potential for AI integration in automated screening after conducting multicentric large-scale studies.

Utility of Perl's Prussian Blue Stain in Exfoliated Buccal Cells of Thalassemia and Sickle Cell Anemia Patients and Their Correlation With Serum Ferritin Levels.

Introduction Iron is essential for all living beings. Excess iron, on the other hand, is dangerous because it causes the creation of free radicals. As a result, iron absorption is carefully managed to maintain a balance between absorption and iron loss in the body. Due to the lack of particular excretory channels for iron in humans, iron excess in the tissues is common. It can be caused by a number of factors, including increased iron absorption, as seen in hemochromatosis, or frequent parenteral iron treatment, as seen in thalassemia and sickle cell anemia patients (a transfusional overload). Aim The study aims to demonstrate Perl's Prussian blue stain to identify iron overload at a preliminary stage and correlate with serum ferritin levels in patients with thalassemia and sickle cell anemia who frequently receive blood transfusions. Materials and methods The present study comprised 62 confirmed cases of thalassemia and sickle cell anemia patients undergoing repeated blood transfusions of a minimum of 15/more, along with 62 clinically healthy individuals between December 2016 and November 2018. The patients with thalassemia and sickle cell anemia were confirmed by hemoglobin electrophoresis (Bio-Rad D-10, Bio-Rad Laboratories, Inc, California, United States). The buccal smears were obtained from these patients along with the controls, and these slides were stained by Perl's Prussian blue stain and were examined under a light microscope. Results Sixty-two cases and 62 controls were considered in the current investigation. Forty-seven of the 62 people had thalassemia, and 15 had sickle cell anemia. Thirty-nine out of the 47 patients with thalassemia and six of the 15 individuals with sickle cell anemia had positive results for Perl's Prussian blue stain. All patients had elevated blood ferritin levels, with varying ranges associated with positive results for Perl's Prussian blue stain. Conclusion The objective of this study was to demonstrate the utility of oral exfoliative cytology in thalassemia and sickle cell anemia patients who often receive blood transfusions as a screening and diagnostic tool. The exfoliative cytology methods' acceptability and simplicity, along with their correlation with serum ferritin levels and Perl's Prussian blue reaction, make this noninvasive procedure an excellent screening and diagnostic tool for all patients who receive repeated blood transfusions.

Liposarcoma Involving Serous Fluid Cavities-A Case Series Illustrating Clinical Implications and the Diagnostic Role of Exfoliative Cytology.

Introduction: Cytological diagnosis of sarcomas requires detailed cytomorphological assessment and integration of immunocytochemistry and/or molecular testing. The role of exfoliative cytology, as compared to aspiration cytology, is less understood. This case series describes well-differentiated/dedifferentiated liposarcomas in effusions, with cytomorphological features, ancillary test results and clinical outcomes detailed. Methods: A computerized search of the department pathology archives was performed for sarcomatous effusions with histological diagnosis or clinical history of well-differentiated/dedifferentiated liposarcoma. Clinical progress, cytology slides, immunocytochemistry and molecular test results were reviewed. Results: Six patients were identified. In 5 patients with clinical follow up, 4 (80%) were deceased within 5 months of malignant effusion. One patient was alive with 12 years disease-free survival after radical resection with adjuvant radiotherapy. Three patients showed dedifferentiation on histology, and high-grade (dedifferentiated) tumor cells were present in effusion cytology of 2 patients. Two showed well-differentiated components only on biopsy, but high-grade (dedifferentiated) tumor cells were identified in cytology. The high-grade tumor cells displayed marked nuclear irregularity, enlargement, size variation, with macronucleoli and multinucleation. Well-differentiated lipomatous components were demonstrated in 4 patients (66.7%), comprising of multivacuolated lipoblasts and atypical lipocytes. CDK4 and MDM2 immunoreactivity in all 3 cases with cell blocks, and CDK4 and MDM2 amplification in one were successfully demonstrated. Conclusion: Lipomatous and dedifferentiated components can be sampled and cytomorphologically identified on effusion fluids of liposarcomas, with sufficient cellularity for immunocytochemistry and molecular testing. Although generally associated with poor prognosis, long disease-free survival with sarcomatous effusion is possible with radical surgery and adjuvant treatment.

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.

Micronuclei detection in oral cytologic smear: does it add diagnostic value?

BACKGROUND: Screening and early diagnosis of oral squamous cell carcinoma (OSCC) are directly associated with increased survival rate and improved prognosis. Noninvasive diagnostic tools have been implemented in the early detection as toluidine blue staining, optical imaging, and oral cytology. This study aimed to assess and compare the presence of micronuclei (MN) in oral exfoliative cytology of healthy controls, subjects exposed to high-risk factors for oral cancer, subjects with oral potentially malignant lesions (OPMLs), and those with malignant oral lesions. SUBJECTS AND METHODS: A total number of 92 subjects were divided into 46 healthy controls with no oral mucosal lesions (23 with no evidence of cancer risk factors and 23 with cancer risk factors), 23 with OPMLs and 23 with oral malignant lesions. All the 92 participants were subjected to cytological sampling for detection of MN. The final diagnosis of the oral lesions was confirmed by the histopathological picture and compared to the cytological results. RESULTS: The results showed that the diagnostic accuracy of MN was higher in OPMLs group (95.2%). The sensitivity of MN test in malignant group was much lower (52.2%); however, all the cytological criteria of malignancy were markedly detected as compared to the OPMLs group. CONCLUSIONS: Conventional oral cytology supported by MN is highly beneficial as adjunctive tool in the screening for early detection of dysplastic oral lesions.

Cytomorphometric Analysis of Exfoliated Buccal Mucosal Cells and DNA Repair Proficiency in Peripheral Lymphocytes in Type 1 Diabetes Mellitus.

The aim of this study was to analyze the cytomorphometric changes in the buccal mucosal smears and to correlate the cytomorphometric findings and glycosylated hemoglobin (HbA1c) in type 1 diabetic (T1D) patients. In addition, DNA repair proficiency in peripheral lymphocytes of T1D patients was evaluated. Smears from the clinically normal buccal mucosa of 46 healthy subjects as controls and randomly selected 45 T1D patients, were stained using the Papanicolaou method, and the mean values (µm2) of nuclear area (NA), cytoplasmic area (CA), and cytoplasmic: nucleus ratio (CNR) were obtained for each patient. DNA damage in diabetics and control subjects was tested on peripheral blood samples using an in vitro mutagen sensitivity assay. Statistical analysis included Student's t-test, one-way ANOVA, and Pearson's correlation test to correlate cytomorphometric changes with HbA1c levels in both T1D and healthy control groups. The mean NA and CA values were higher in T1D while the mean CNR was found to be higher in the healthy control group. Statistical significance between the two groups was found only for the mean NA (P < 0.001) and mean CNR values (P < 0.001). A significant correlation was seen between HbA1c and CNR in both the T1D group (P = 0.015) and the healthy control group (P = 0.02). Significant differences were noted between the T1D and control subjects with regard to the frequency of damaged cells and DNA repair proficiency (P < 0.05). Significant cytomorphometric alterations were detected in the buccal mucosal cells of diabetic patients, which may aid in the early detection of diabetes or predict the risk for future disease complications among T1D patients.

Endometrial Aspiration Cytology: A Cross-Sectional Comparative Study of Its Efficacy and Sensitivity in Diagnosing Gynecological Disorders.

Introduction Endometrial aspiration cytology (EAC) is a noninvasive, rapid, and cost-effective procedure for diagnosing gynecological disorders. This study aimed to validate endometrial aspiration as a routine, safe, and efficient outpatient diagnostic procedure, correlating its findings with histopathology evaluations to facilitate early surgical planning for patients with abnormal uterine bleeding. Materials and methods This cross-sectional study involved patients of reproductive, menopausal, and postmenopausal age groups who presented with diverse gynecological concerns that required dilatation and curettage. Endometrial aspiration was performed using a Karman cannula (Angiplast Pvt. Ltd., Vatva, India), and the obtained material was prepared into smears and stained for evaluation. Sampling adequacy, the nature of glandular and stromal cells, phasing of the endometrium, and other abnormalities were assessed and correlated with histology to examine the diagnostic utility of endometrial cytology. Results EAC showed 90.66% sampling adequacy with the Karman cannula. The sensitivity of EAC for diagnosing benign and malignant conditions was 88.7% and 100%, respectively. Conditions including secretory endometrium, proliferative phase, tuberculous endometritis, and glandular hyperplasia were diagnosed using EAC and confirmed by histopathology. Six malignancies were successfully diagnosed on cytology smears, while challenges in differentiation and sampling errors were recognized as limitations of the technique. Conclusions This study established EAC as a highly sensitive, minimally invasive preliminary diagnostic tool for gynecological disorders, particularly effective in diagnosing malignancy. Despite certain limitations, the procedure's ease, cost-effectiveness, and safety underscore its potential for routine use by surgeons.

Periodic acid-Schiff staining in oral exfoliative cytology of diabetic patients: The odyssey for noninvasive screening - A systematic review and meta-analysis.

Background: Diabetes mellitus is a dominant metabolic disorder in the current fast paced culture; its prevalence is escalating worldwide and among all age groups. Guidelines recommend universal screening for diabetes; however, the uptake of these tests in individuals suggests challenges owing to invasive nature of blood collection. Thus, there arises the need for a noninvasive investigation for diabetes screening with a minimum paraphilia and for all medical settings. Materials and Methods: We have thus conducted a systematic review and meta-analysis according to Preferred Reporting Items for Systematic Reviews and Meta-Analysis guidelines, to quantify the association between diabetes and effectiveness of periodic acid-Schiff staining of exfoliative cytology as a screening method. MEDLINE, EMBASE, Cochrane, Scopus, Web of science, CINAHL, and Google Scholar were searched systematically for publications dated till July 20, 2022. Six studies (case-control cross-sectional) were selected and evaluated in depth. The statistical evaluation was done with a forest plot with odds ratio and weightage examined. Results: It was proved that exfoliative cytology with periodic acid-Schiff (PAS) staining can be used as a screening test for diabetic status evaluation. The findings suggest that the aforesaid noninvasive test is a conclusive screening practice for diabetics. Conclusion: This systematic review and meta-analysis suggest that PAS staining in exfoliative cytology can be used as a noninvasive screening in diabetic individuals to assess the current level of blood glucose. Given the increased risk of diabetes worldwide, higher quality prospective evidence is suggested in larger sample sizes with other metabolic disorders, ethnicity, and oral disorders to further evaluate the plausibility of the results.

Exfoliated oral mucosa cells as bioindicators of short- and long-term systemic titanium contamination.

BACKGROUND: Humans are exposed to exogenous sources of titanium-containing particles that can enter the body mainly by inhalation, ingestion, or dermal absorption. Given the widespread use of biomaterials in medicine, the surface of a titanium (Ti) biomedical device is a potential endogenous source of Ti ions and/or Ti-containing particles, such as TiO2 micro-(MPs) and nano-particles (NPs), resulting from biotribocorrosion processes. Ti ions or Ti-containing particles may deposit in epithelial cells of the oral mucosa, and the latter may therefore serve as bioindicators of short and long-term systemic Ti contamination. The aim of the present study was to histologically and quantitatively evaluate the presence of Ti traces in cells exfoliated from the oral mucosa as possible bioindicators of systemic contamination with this metal at short and long-term experimental time points METHODS: Thirty Wistar rats were intraperitoneally injected with a suspension of titanium dioxide (TiO2) (0.16 g/100 g body weight of TiO2 in 5 ml of NaCl 0.9%) using 5 nm NPs (Group: TiO2-NP5; n = 10), 45 µm MPs (Group: TiO2-MP45; n = 10), or vehicle alone (Control group; n = 10). At one and six months post-injection, right-cheek mucosa cells were obtained by exfoliative cytology using a cytobrush; they were spray fixed and stained using Safranin or the Papanicolaou technique. The smears were cytologically evaluated (light microscopy) to determine the presence of particulate material, which was also analyzed microchemically (SEM-EDS). Left-cheek mucosa cells were similarly obtained and re-suspended in 5 ml of PBS (pH: 7.2-7.4); the samples corresponding to each group were pooled together and analyzed spectrometrically (ICP-MS) to determine Ti concentration in each of the studied groups. Blood samples were obtained for histological determination of the presence of particulate material on Safranin-stained blood smears and determination of plasma concentration of Ti by ICP-MS RESULTS: Different size and shape metal-like particles were observed inside and outside epithelial cells in TiO2-NP5 and TiO2-MP45 cytological smears at both one and six months post-injection. EDS analysis showed the presence of Ti in the particles. ICP-MS revealed higher Ti concentrations in both TiO2 injected groups compared to the control group. In addition, Ti concentration did not vary with time or particle size. Monocytes containing particles were observed in blood smears of TiO2-exposed animals one- and six-months post-injection. Plasma levels of Ti were significantly higher in TiO2-NP5- and TiO2-MP45- exposed animals than in controls (p < 0.05), and Ti concentration was significantly higher at one month than at six months in both TiO2-exposed groups (p < 0.05). CONCLUSIONS: Cells exfoliated from the oral mucosa could be used as bioindicators of short- and long-term systemic contamination with Ti. Exfoliative cytology could be used as a simple, non-invasive, and inexpensive diagnostic method for monitoring biotribocorrosion of Ti implants and patient clinical follow-up.

Correlation of the activities of antioxidant enzymes superoxide dismutase and glutathione reductase with micronuclei counts among areca nut chewers of Manipuri population using exfoliative cytology: A preliminary study.

Context: Areca nut (AN) is a potent cytotoxic and genotoxic agent. Oxidative stress-induced by chewing of AN can cause DNA damage leading to nuclear anomalies such as micronuclei (MN) and also alters antioxidant defense mechanisms, leading to genomic instabilities and the development of oral cancer. Aims: The aim of this study is to study the correlation between the levels of glutathione reductase (GR) and superoxide dismutase (SOD) in exfoliated buccal mucosal cells and the genotoxicity levels (MN count) in chronic AN chewers. Materials and Methods: The present study was conducted with the approval of the Research Ethics Board in 60 individuals; 40 cases (Group I-20 raw AN chewers, Group II-20 dried areca with tobacco chewers), and 20 controls as Group III in the age group of 18-68 years who attended the outpatient department of our college. Estimation of SOD and GR and MN assessment was done using buccal exfoliated cells. Statistical Analysis Used: Statistical analysis was performed using the Student's t-test and one-way ANOVA. Results: Antioxidant levels were found to be significantly reduced in both Group I and Group II in comparison to the control group. Group II showed significantly reduced level of GR in comparison to Group I. The MN count was significantly increased in Group II in comparison to Group I. The MN counts showed an inverse correlation to the activities of the antioxidant enzymes. Greater activities of antioxidant enzymes correlated with decreased MN counts. Conclusions: Detection of MN in AN chewers with or without tobacco can be a useful biomarker for clinical screening procedures that may be used as a risk marker for oral cancer. It is important to increase the awareness programs to educate the public about the deleterious effects of AN chewing, emphasize on early intervention of AN chewing habit and thus prevent the development of oral cancer.

Analysis of false-negatives in exfoliative cytology in oral potentially malignant disorders: A retrospective cohort study.

INTRODUCTION: Keratinized lesions have been a conceivable false-negative (FN) factor in oral exfoliative cytology (OEC); however, other factors are poorly analyzed. In this study, we aimed to identify the factors influencing the accuracy of OEC and FNs focusing on the lesion characteristics, patient background, and surgeon factors in oral potentially malignant disorders (OPMD). MATERIAL AND METHODS: We retrospectively studied 44 patients who underwent both OEC and histopathological diagnosis. Sensitivity, specificity, FN rate, false-positive (FP) rate, and prevalence of both methods were compared. Similarly, accuracy indices were compared among clinical diagnosis groups (leukoplakia vs. other diagnosis). The association between patient and surgeon-related factors influencing FN OEC results were investigated using Fisher's exact test and a multiple logistic regression analysis. RESULTS: Overall, the sensitivity; specificity; and FN, FP, and prevalence rates of OEC were 31.8%, 82.1%, and 68.8%, 17.9%, and 36.4%, respectively. Leukoplakia was significantly more common in clinical diagnosis (P = 0.007) with sensitivity, specificity, and FN rates of 20.0%, 95.2%, and 80.0%, respectively. Contrarily, non-keratinized lesions had sensitivity, specificity, and FN of 83.3%, 85.7%, and 16.7%, respectively. In the prevalent group, leukoplakia and anucleate squamous cells were significantly associated with FN cases (P = 0.013, P = 0.050). On multivariate analysis in OEC negative patients, age ≤64 (P = 0.050) and location on the tongue (P = 0.047) was independently associated with FNs. CONCLUSION: FN of OEC was conceivable to be due to poor deep-seated cell sampling, which was associated with leukoplakia, age, and location. Therefore, these factors may be considered in the evaluation of OEC results.

Cytomorphometric Evaluation of Oral Mucosa of Patients with Duchenne Muscular Dystrophy.

Duchenne muscular dystrophy (DMD) is a rare genetic disorder in which patients progressive muscle deterioration and low life expectancy. Since DMD has no cure, treatment slows the deterioration process using corticosteroids and other medications, including oral bisphosphonates (BP). These drugs can lead to toxicities in both soft and hard tissues. The objective of this study was to present cytological changes in DMD patients, comparing to young, healthy individuals. Cytological smears were obtained from buccal mucosa from nine DMD patients and five healthy patients. The Papanicolaou technique was used to stain the slides. Images were captured using a photomicroscope, and 50 clearly defined cells were selected. The size and ratio between the nucleus and cytoplasmic areas (NA/CA) diameter were measured using a computer program. All the DMD patients presented a statistical difference (p < 0.0001) in the size of the cytoplasm (0.60 ± 0.33 µm) and the NA/CA ratio (0.17 ± 0.07 µm) was seen when compared to the control group (cytoplasm 0.60 ± 0.15 µm and nucleus 0.14 ± 0.05 µm). Furthermore, in one DMD in use of BP who presented Medication-Related Osteonecrosis of the Jaw (MRONJ), presented higher differences in the cytoplasmic (1.20 ± 0.65 µm) and nuclear (0.20 ± 0.12 µm) sizes (p < 0.0001) compared with all others DMD patients. These results suggest that the drugs used or the systemic condition of individuals with DMD may contribute to these changes.

Cytological Screening Model of Normal Oral Mucosa Exposed to Carcinogens: A Pilot Study.

INTRODUCTION: Oral cytopathology is able to detect incipient cellular alterations, but it is not routinely applied to this purpose. We aimed to establish a model to screen individuals with no oral lesion exposed to smoking/alcohol, by means of the nuclear area, cell proliferation rate, and analysis of genetic damage. METHODS: In this cross-sectional pilot study, 90 patients were allocated into 3 groups: oral cancer group (patients with oral squamous cell carcinoma), tobacco/alcohol group (patients without oral lesions and exposed to these risk factors), and control group (individuals with no lesion and not exposed to tobacco and alcohol). The cytological smears performed in these individuals were stained with Papanicolaou, a silver-staining and a Feulgen reaction. The nuclei of cells were measured, and AgNORs/nucleus and micronuclei (MN) were quantified. The cutoff values were stipulated evaluating the healthy mucosa (control group) and the cancerization field mucosa (oral cancer group). RESULTS: Cutoff values for the screening of individuals exposed to carcinogens were ≥8% of nuclei larger than 100 µm2, ≥3.38 AgNOR/nucleus, and ≥3 MN per 1,000 cells. CONCLUSIONS: Nuclear area measurement and AgNORs/nucleus and MN quantification identified the incipient phase of oral carcinogenesis. A screening model for individuals without oral lesion exposed to smoking/alcohol was proposed.

Cytomorphometric analysis of squames obtained from normal mucosa, leukoplakia and oral squamous cell carcinoma.

INTRODUCTION: Exfoliative cytology is becoming increasingly important in the early diagnosis of oral cancers, as a procedure for obtaining cell samples, which can be analyzed by sophisticated diagnostic techniques. Quantitative techniques, based on the evaluation of parameters such as nucleus area, cytoplasm area and nucleus-to-cytoplasm area ratio, may increase the sensitivity of exfoliative cytology for early diagnosis of oral cancers, since these techniques are precise, objective and reproducible. OBJECTIVES: This study was undertaken to employ the cytomorphometric quantitative techniques to determine the value of exfoliative cytology and to assess this method to analyze the effectiveness in providing additional diagnostic test for the detection of early oral malignancy. MATERIALS AND METHODS: The study consisted of three groups. The experimental group comprised of 20 subjects with oral leukoplakia and 20 subjects with oral squamous cell carcinoma of the buccal mucosa. The control group comprised of 10 apparently healthy subjects. They were subjected to clinical examination and cytosmear of the buccal mucosa. A standard wooden tongue spatula/metal spatula moistened with normal saline was used to obtain scrapings of buccal mucosa. The scrapings were spread on plain glass slides and immediately fixed in Biofix spray, followed by staining with the Papanicolaou technique. Then, the microscopic pictures were captured onto a computer and are cytomorphometrically analyzed using an image analyzer. Since this study involved multiple groups, one-way analysis of variance (ANOVA) was used for comparing the parameters for multiple groups. Where significant difference was seen, the ANOVA test was followed by Mann-Whitney test, for pairwise comparison. RESULTS: The mean count of nuclear area and nuclear diameter showed an increase from Group I to Group II, Group I to Group III and Group II to Group III, which was statistically significant. The mean count of cell area and cell diameter showed a decrease from Group I to Group II, Group I to Group III and Group II to Group III, which was statistically significant. The mean count of nuclear/cytoplasm area ratio showed an increase from Group I to Group II, Group I to Group III and Group II to Group III, which was statistically significant. CONCLUSION: With the advancements in the field of quantitative exfoliative cytology, interest in oral cytology has once again emerged in the diagnosis of oral premalignancy and malignancy. Computer-aided analysis with the sophisticated software increases the speed and accuracy of cytological measurements, which are repeatable. These versatile systems facilitate quantitative oral cytological assessments, which may become a viable procedure for the early detection of oral premalignancy and malignancy.

Evaluation of Role of Visual Inspection Using Acetic Acid (VIA) and Exfoliative Cytology in Screening and Early Detection of Oral Premalignant Lesions and Oral Cancer.

OBJECTIVE: Oral cancer accounts for 50-70% of total cancer mortality. VIA screening has drastically changed the morbidity and mortality related to cervical cancer. In present study, we assessed the role of 5% Acetic Acid as a vital stain in oral mucosa of chronic tobacco chewers, to know if it can help in early diagnosis and improved prognosis of oral malignancies and to assess the sensitivity and specificity of VIA and exfoliative cytology for screening of Oral premalignant and malignant lesions in high risk population with limited health care infrastructure. METHODS: This was an outpatient department based prospective study done in a tertiary hospital over a period of two years. 150 cases with a history of chronic tobacco use were evaluated. Visual inspection (VI) followed by VIA using 5% Acetic acid was done. Oral scrapes were taken for cytological examination followed by biopsy for histopathological evaluation, wherever possible. RESULTS: Sensitivity, specificity, PPV, NPV and accuracy of VIA and Exfoliative cytology was 71%, 65%, 83%, 48%, 69% and 98%, 65%, 89%, 92%, 88% respectively keeping histology as gold standard. CONCLUSION: Acetic acid along with exfoliative cytology can be used as simple, cost effective and convenient methods for mass screening of OPMDs and OSCC in high risk population where biopsy is not possible.
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The ocular pyogranulomatous lesion in a Gentoo penguin (Pygoscelis papua) from the Antarctic Peninsula: evaluation of microbiological and histopathological analysis outcomes.

In this study, it was aimed to present the results of microbiological, cytological, histopathological, and immunohistochemical analyses of ocular samples from an Antarctic (Ardley Island, King George Island) Gentoo penguin chick (Pygoscelis papua) with a pyogranulomatous lesion in the right eye. Samples were taken from both the healthy left eye and the lesion in the right eye. Conventional culture methods and phenotypic and molecular tests were used for bacterial isolation and identification, respectively. None of the isolates could be identified phenotypically. As a result, four of the five isolates obtained from the right eye were considered to belong to putative novel bacterial species and taxa as their similarity to GenBank data was below 98.75%. The isolates were considered to be Pasteurellaceae bacterium, Corynebacterium ciconiae, Cardiobacteriaceae bacterium, Actinomyces sp., and Dermabacteraceae bacterium. The only isolate from the left eye was identified as Psychrobacter pygoscelis. The cytological analysis demonstrated cell infiltrates composed mostly of degenerate heterophils, reactive macrophages, plasma cells, lymphocytes, and eosinophils. Based on histopathological findings, the lesion was defined as a typical pyogranulomatous lesion. Immunohistochemistry demonstrated that the granuloma was positive for TNF-α, IL-4, MMP-9, IL-1ß, and IL-6. This is the first documented report of the unilateral pyogranulomatous ocular lesion in a Gentoo penguin chick, living in its natural habitat in Antarctica. This report also describes the isolation of four bacteria from the infected eye, which are considered to belong to novel Genus, species, or taxa. The primary bacterial pathogen that caused the ocular lesion was not able to be detected and remains unclear.

A Cytological Study Enlightening the Unseen Effects of Concomitant Chemoradiotherapy in Contralateral Normal Buccal Mucosa of Oral Squamous Cell Carcinoma Patients.

Background/objectives In patients receiving concomitant chemoradiotherapy (CCRT) as a treatment for oral squamous cell carcinoma (OSCC), cytological changes were seen not only in neoplastic epithelial cells but the non-neoplastic epithelial cells are also affected, resulting in cytopathological atypical changes. The present study was designed to observe oral epithelial atypical cytopathologic changes induced in contralateral normal buccal mucosa in OSCC patients receiving CCRT. Methods The study included 150 patients with OSCC treated by CCRT whose details were collected from the Institute of Nuclear Medicine and Oncology Lahore (INMOL) Hospital Lahore. Cytological smears were obtained from the contralateral normal buccal mucosa of OSCC patients. The serial scrape smears were taken before, immediately after, on the 17th day (mid of treatment), and at the end of CCRT, whereas 20 patients were taken as normal healthy controls and were not exposed to CCRT. The smears were stained with hematoxylin and eosin and Papanicolaou stain. SPSS version 20 (Armonk, NY: IBM Corp.) was used for statistical analysis and p > 0.05 was considered to be significant. Results CCRT-induced oral epithelial atypical cytological changes were predominantly noted at end of therapy (19.7%) in the contralateral normal buccal mucosa. Nuclear atypia features were higher on the 17th day and end of treatment; whereas, epithelial atypia was mainly observed on the 17th day of CCRT (40%). A highly significant association was observed between epithelial atypia and radio-chemotherapy dose (p = 0.045), between CCRT-induced epithelial atypical cytological changes and days of treatment (p = 0.001), and between days of CCRT and nuclear atypia (0.000) accordingly. Atypia was not observed in any control group. Conclusion Varying degrees of oral epithelial atypical cytological changes may occur in otherwise normal contralateral mucosa of the patients receiving CCRT.

Lymphoreticular malignancies in serous effusions: Cytomorphologic, flow cytometric and immunocytochemical analysis.

INTRODUCTION: Involvement of body fluids by lymphoreticular malignancies (LRM) is rare and often associated with poor prognosis and decreased overall survival. The present study was conducted to analyze the characteristic cytomorphologic, flow cytometric and immunocytochemical features of LRMs in serous effusions. MATERIALS AND METHODS: This was a three-year retrospective study. A total of 218 effusion samples, reported as involved by lymphoreticular malignancies, on cytology, were reviewed. All the cases wherein the cytological diagnosis was confirmed by flow cytometric (FCM) and/or immunocytochemical (ICC) studies were retrieved and studied in detail. FCM and/or ICC were performed in a total of 51/218(23.4%) samples, including 30 pleural (58.8%), 18 peritoneal (35.3%), and 3 pericardial fluid (5.9%) samples. RESULTS: The cytomorphologic diagnoses included infiltration by non-Hodgkin lymphoma (NHL;n = 27), infiltration by LRM (n = 19), infiltration by chronic lymphocytic leukemia (CLL;n = 2), Hodgkin's lymphoma (HL;n = 1) and suggestive of infiltration by LRM (n = 2). FCM and/or ICC confirmed the diagnoses as infiltration by T-cell lymphoblastic lymphoma in 18; mature B-cell NHL in 10; Burkitt lymphoma in 7; diffuse large B-cell lymphoma in 4; follicular lymphoma, T- cell NHL and CLL in 2 samples each and hairy cell leukemia, plasmablastic lymphoma and HL in 1 sample each. 94.1% concordance was noted between the initial and final cytologic diagnosis. CONCLUSIONS: Involvement of body fluids and effusions by LRMs, though rare, carries an immense prognostic significance and hence the prompt detection is crucial. Detection of these malignancies by cytologic examination of effusions is challenging yet potentially useful and the least invasive method available to establish an early diagnosis.