Cytology Research Articles

Safety and Effects of Intravaginal Administration of Lacticaseibacillus rhamnosus CRL1332 Immobilized on Nanofibers in a Murine Experimental Model

The design of probiotic hygiene products for daily use is considered an adequate alternative for the restoration of the vaginal microbiome, maintaining health, and/or preventing infections of the female urogenital tract. Most of these probiotic products are available on the world market, but their efficacy and safety are not sufficiently documented. One of the requirements to transfer novel probiotic formulas/products to the productive sector is to demonstrate their innocuity and the absence of adverse or collateral effects on the host, mainly assayed in experimental models. The inclusion of beneficial lactobacilli in nanofibers by electrospinning technique has shown promising application possibilities, and the immobilization of Lacticaseibacillus rhamnosus CRL1332 in nanofibers with and without bioprotective substances and their characterization were previously performed by our research group. In this work, the safety of the intravaginal (i.va.) administration of these functional nanofibers in a murine experimental model was evaluated. L. rhamnosus CRL1332 immobilized in different nanofibers was intravaginally inoculated into mice (seven daily doses). Vaginal washes were taken for microbiological (cultivable lactobacilli) and cytological techniques, and the vagina was used for histological and morphological-ultrastructural evaluation. Our results demonstrated that the intravaginal administration of L. rhamnosus CRL1332 immobilized in nanofibers is safe in murine models, given the absence of an inflammatory response at the cytological and histological levels, with minor modifications at the ultrastructural level, and also related to the normal cultivable vaginal microbiota. On the other hand, the number of cultivable lactobacilli increased in the vagina of mice receiving L. rhamnosus CRL1332 nanofibers. The results indicate the safety of lactobacilli-functional nanofibers and support their inclusion in the design of vaginal probiotic products to prevent/treat urogenital infections and reconstitute the women’s vaginal microbiota.

Cytology and Histology of Thyroid Nodules: Exploring Novel Insights in the Molecular Era for Enhanced Patient Management.

Significant advancements have been made over the past decade in our understanding of thyroid cancers, encompassing histomorphology, cytology, and ancillary techniques, particularly molecular tests. As a result, it is now feasible to put forth a comprehensive histo/cytomolecular approach to treating these tumors, thereby offering patients treatments that are precisely tailored to their unique circumstances.

Influence of sexually transmitted infections on the cervical cytological abnormalities among Iranian women: A cross-sectional study.

Sexually transmitted infections (STIs) are one of the world's most severe health challenges. The existence of STIs such as human papillomavirus (HPV) might cause cervical cell changes leading to cervical cancer. This study aims to assess the association of STIs with cervical cytological abnormalities and genital warts among women in northeastern Iran. This cross-sectional study was carried out on 190 women referred to the central laboratory of Academic Center for Education, Culture, and Research, Mashhad, Iran from March to July 2022. The presence of genital infections caused by Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, and Herpes simplex viruses (1 and 2) were assessed using the real-time polymerase chain reaction method. HPV genital infection was detected based on the principles of reverse hybridization, and cellular changes in the cervix were examined by the liquid-based cytology technique. The mean age of participants was 35.33 8.9 yr. 34 different HPV genotypes were detected in all HPV-positive cases, and the most common genotype was low-risk HPV6. No significant association was found between STIs and cervical cytology abnormalities. The prevalence rates of sexually transmitted pathogens among HPV-positive and HPV-negative individuals were 10.9 and 1.6%, respectively. The frequency of genital warts was significantly higher in cases with multiple infections of high- and low-risk HPV genotypes. High percentages of the participants with non-HPV STIs and HPV infection had normal cervical cytology. It is advised to use STIs and HPV diagnostic tests along with cytology examinations for cervical cancer screening.

An immersive simulation strategy to teach cytology sample collection methods and basic diagnosis skills: A two academic center study.

Collecting cytology samples and making simple diagnoses are skills taught in veterinary universities, mostly in a passive way. Simulators enhance practical skills learning, increasing student engagement through immersive activities. These strategies have not been carefully assessed in veterinary cytology. This study aimed to describe a simulator developed for training cytology sample collection methods and assess the utility of an immersive simulation strategy to learn and practice the collection of cytology samples. A flipped classroom with a station design was followed. Students first watched video tutorials on sampling collection, listened to brief cases' clinical histories, and moved to immersive simulator stations. There, they practiced imprints, swabs, and fine-needle aspiration (FNA). Microscopic observation of the material was available through augmented reality tools. Students were evaluated by Objective Structured Clinical Examination (OSCE) tools on their ability to perform FNA on stuffed animal models. Students answered two questionnaires addressing their learning outcomes. Second- and third-year students from two centers (n = 129) practiced cytologic collection methods in simulators which significantly changed their willingness to perform FNA in live animals after the class activities. OSCE pass rates over 90% were obtained for most steps of FNA, and students rated the activity as essential/very relevant for learning. Immersive simulation strategies were effective at increasing student comfort with cytologic sampling techniques. This approach should be included in the veterinary curriculum as it can increase the quality of cytology samples and could potentially improve the cytologic diagnosis of a submitted sample.

Role of scrape cytology smear preparation in the diagnosis of ovarian masses-utility and pitfalls.

Scrape cytology technique is useful for rapid intraoperative diagnosis of ovarian mass lesions that acts as an adjunct to frozen section examination. Though ovaries can be accessed by laparoscopy and USG guided FNAC, there are controversial reports regarding the safety of these procedures. The present study has been designed to evaluate the role of scrape cytology in a variety of ovarian mass lesions. To study the cyto-morphology of ovarian mass lesions and to evaluate the role of scrape cytology technique in correctly diagnosing the ovarian lesions taking histopathological diagnosis as gold standard. This is a prospective observational study done on 61 ovarian mass lesions as received from the Obstetrics and Gynecology department of our Institution. Out of 61 cases, 58 (95.08%) cases were correctly diagnosed with respect to categorization and typing. The age ranged from 14 to 65 years with the mean age being 38.1 years. On histopathology, out of 61 cases, 39 (63.93%) were epithelial along with sub categorization of benign, borderline, and malignant, 13 (22.9%) were germ cell tumors, 5(8.19%) were sex cord stromal tumors, 3 (4.91%) were hemorrhagic cysts and the remaining 1 (1.63%) was massive ovarian edema. Thus, on comparing with histopathology, the sensitivity and specificity of scrape cytology technique were 93.55% and 96.67%, and the diagnostic accuracy was 95.08%. Scrape cytology of ovarian lesions can give quick and reliable results. Also, proper training of cytopathologists chiefly regarding the sampling technique, gross presentation of ovarian lesions and interpretation of scrape cytology smears is needed. Further studies to provide standard guidelines and reporting criteria will prove to be helpful.

Usefulness of imprint cytology for early diagnosis in vertebral lesions

Abstract Background: Vertebral pathologies can be divided into degenerative, infective, or neoplastic. Infective and neoplastic lesions can progress rapidly and may lead to compression of the cord. Thus, rapid diagnosis for these lesions is important. Frozen section and fine-needle aspiration cytology techniques are less useful in hard bony tumors. Imprint cytology has been successfully used in the diagnosis of soft tissue tumors. The purpose of this study was to evaluate the role of imprint cytology in the early diagnosis of various spinal pathologies. Materials and Methods: Thirty-eight patients with suspected diagnosis of infection or tumor underwent transpedicular biopsy using Jamshidi needle. At least two imprint slides were made using core tissue obtained from biopsy. These slides were, thereafter, stained with the May-Grunwald–Giemsa method. The core biopsy sample was also sent for histopathological examination, after fixing it with 10% formalin. Paraffin-embedded blocks and slides were made for histopathology reporting, as per institution’s protocol. All imprint cytology slides were screened and reported by same pathologist. Results: The sensitivity of imprint cytology was 68.6%. Specificity, positive predictive value, negative predictive value, and overall accuracy of the imprint cytology were 100%, 100%, 21.4%, and 71%, respectively, when compared to histopathology. The mean duration of imprint cytology reporting was only 36 min while that for histopathological report was 15 days. Conclusion: Imprint cytology is a simple and rapid method for obtaining diagnosis in suspected vertebral lesions of infection or tumor. It is a reliable and cost-effective method in experienced hands. Although imprint cytology shows modest correlation in the ability to identify positive results, it forms a good diagnostic tool in confirming true negative cases with high diagnostic efficacy.

Leukocyte differentiation in bronchoalveolar lavage fluids using higher harmonic generation microscopy and deep learning.

In diseases such as interstitial lung diseases (ILDs), patient diagnosis relies on diagnostic analysis of bronchoalveolar lavage fluid (BALF) and biopsies. Immunological BALF analysis includes differentiation of leukocytes by standard cytological techniques that are labor-intensive and time-consuming. Studies have shown promising leukocyte identification performance on blood fractions, using third harmonic generation (THG) and multiphoton excited autofluorescence (MPEF) microscopy. To extend leukocyte differentiation to BALF samples using THG/MPEF microscopy, and to show the potential of a trained deep learning algorithm for automated leukocyte identification and quantification. Leukocytes from blood obtained from three healthy individuals and one asthma patient, and BALF samples from six ILD patients were isolated and imaged using label-free microscopy. The cytological characteristics of leukocytes, including neutrophils, eosinophils, lymphocytes, and macrophages, in terms of cellular and nuclear morphology, and THG and MPEF signal intensity, were determined. A deep learning model was trained on 2D images and used to estimate the leukocyte ratios at the image-level using the differential cell counts obtained using standard cytological techniques as reference. Different leukocyte populations were identified in BALF samples using label-free microscopy, showing distinctive cytological characteristics. Based on the THG/MPEF images, the deep learning network has learned to identify individual cells and was able to provide a reasonable estimate of the leukocyte percentage, reaching >90% accuracy on BALF samples in the hold-out testing set. Label-free THG/MPEF microscopy in combination with deep learning is a promising technique for instant differentiation and quantification of leukocytes. Immediate feedback on leukocyte ratios has potential to speed-up the diagnostic process and to reduce costs, workload and inter-observer variations.

USE OF CYTOLOGICAL SMEAR AND CELL BLOCK TECHNIQUE FOR THE DIAGNOSIS OF PLASMA CELL MYELOMA INFILTRATION IN THE MANDIBLE

Fine-needle aspiration cytology (FNAC) is a diagnostic tool using a needle to collect samples for cytopathological examination. We report a case of a 49-year-old male patient with previous medical history of plasma cell myeloma (PCM), who had osteolytic lesions in the mandible, diagnosed by FNAC. Imaging exams exhibited multiple osteolytic lesions with ill-defined margins in the bilateral mandibular body. FNAC of the intraosseous lesion of the mandible was performed. Cytological smear and cell block technique revealed plasma cells in different degrees of maturation, predominantly eccentric pleomorphic nuclei and abundant, eosinophilic cytoplasm without a paranuclear halo. Immunohistochemistry exhibited positivity for CD138, lambda light chain and Ki-67 (proliferation index was ∼50%). Based on clinicoradiographic, microscopic, and biochemical findings, the diagnosis was PCM infiltration in the mandible. The patient underwent chemotherapy and autologous hematopoietic stem cell transplantation, and had partial remission after 10 months of follow-up. Fine-needle aspiration cytology (FNAC) is a diagnostic tool using a needle to collect samples for cytopathological examination. We report a case of a 49-year-old male patient with previous medical history of plasma cell myeloma (PCM), who had osteolytic lesions in the mandible, diagnosed by FNAC. Imaging exams exhibited multiple osteolytic lesions with ill-defined margins in the bilateral mandibular body. FNAC of the intraosseous lesion of the mandible was performed. Cytological smear and cell block technique revealed plasma cells in different degrees of maturation, predominantly eccentric pleomorphic nuclei and abundant, eosinophilic cytoplasm without a paranuclear halo. Immunohistochemistry exhibited positivity for CD138, lambda light chain and Ki-67 (proliferation index was ∼50%). Based on clinicoradiographic, microscopic, and biochemical findings, the diagnosis was PCM infiltration in the mandible. The patient underwent chemotherapy and autologous hematopoietic stem cell transplantation, and had partial remission after 10 months of follow-up.

Development of a postoperative nomogram to predict disease-specific mortality in gastric cancer: A competing risk analysis using variables the AJCC recommends be collected and registered.

e16099 Background: The American Joint Committee on Cancer (AJCC) has increasingly recognized the need for individual risk prediction model for the era of tailored therapy. In several reports, gastric cancer postoperative nomograms for predicting overall or disease-specific survival have been described. The AJCC Precision Medicine Core introduced the attractiveness of disease-specific mortality (DSM) as an endpoint of risk model. This study aimed to develop the first pretreatment gastric cancer postoperative nomogram for predicting DSM that considers competing risks (CRs). Methods: The nomogram was developed using data of 2,501 patients with primary gastric cancer who are potential candidate for adjuvant chemotherapy based on NCCN guideline, and it was created with a multivariable Fine and Gray subdistribution proportional hazard regression model. Fifteen pathological or non-tumor-related variables (age, sex tumor location, tumor size, macroscopic type, histology, depth (pT), number of positive nodes (pN), number of negative nodes, location of positive nodes, lymphovascular invasion, lavage cytology (CY), tumor margin, serum CEA and serum CA19-9) were collected. They are recommended to collect and register by the AJCC. The model was validated internally using measures of discrimination (Harrell’s C-index), calibration and decision curve analysis. Results: In the development procedure, multivariable analysis for DSM selected 12 variables (age, tumor location, macroscopic type, histology, pT, pN, number of negative nodes, location of positive nodes, lymphovascular invasion, CY, tumor margin and serum CEA) for constructing the nomogram. The developed nomogram showed good discrimination, with a C-index of 0.802(0.786-0.817) that of the American Joint Committee on Cancer (AJCC) pathological stage was 0.714 (0.697-0.734). The calibration appeared to be accurate for the 5-year prediction. Compared with scenarios in which no prediction model was used for postoperative treatment decision making (i.e., assume all or assume none), the nomograms had a favorable net benefit across a wide range of decision threshold probabilities between 5-year DSM probabilities of approximately 5 and 95%. Conclusions: This new postoperative risk model accurately predicts DSM in gastric cancer and can be used for patient counseling in clinical practice.

Prognostic Value of Peritoneal Lavage Cytology in Potentially Resectable Pancreatic Cancer Stratified by Cytologic Status.

Patients with pancreatic ductal adenocarcinoma (PDAC) with positive peritoneal lavage cytology (CY) reportedly have poor prognoses. However, the value of diagnosis of suspicious for malignancy on CY is unknown. This study aimed to elucidate the prognostic impact of CY by focusing on CY subgroups. Data were collected from 231 resectable PDAC patients who underwent curative-intent resection. Patients were divided into three CY-based groups: negative (CY0), suspicious for malignancy (CY-S), and positive (CY1). Clinicopathological characteristics and prognostic factors were analyzed. CY1 and CY-S were diagnosed in 7.8% and 3.9% of the patients, respectively. The CY1 group had significantly larger tumors and higher frequencies of distal tumors, anterior pancreatic tissue invasion, retropancreatic tissue invasion, and R1 resection than the CY0 group. Patient characteristics did not differ between the CY0 and CY-S groups. The CY1 group exhibited worse survival than the CY0 and CY-S groups (median survival time: 18.8 vs. 39.6 months, p=0.0021 and vs. 62.2 months, p=0.018). Multivariate analysis for survival indicated that a tumor size >2 cm, preoperative CA19-9 value >100 U/ml, CY1, lymph node metastasis, R1 resection, and lack of adjuvant chemotherapy were associated with poor prognosis. Both the CY1 and CY-S groups had higher frequencies of peritoneal recurrence than the CY0 group (50% vs. 11.8%, p<0.001 and 44.4% vs. 11.8%, p=0.019). The prognosis of the CY1 group was poor. Although CY-S was associated with a higher frequency of peritoneal recurrence than CY0, the long-term outcomes of patients with surgical treatment were acceptable.

SENODAY: A New Perspective of Handling Breast Cancer

BackgroundTo evaluate the feasibility of a same day breast cancer diagnosis and management protocol, consequently decreasing time to treatment and immediately reassuring patients with benign diagnosis. Materials and MethodsA total of 60 women underwent breast exam during SENODAY in our cancer center between January 2020 and December 2022. Patients are first seen by a breast surgeon who mentions whether the patient's history and physical exam are suspicious of malignancy. Patients are then sent to the radiologist who performs a complete radiologic assessment, classifies the lesions, and performs a biopsy when necessary. The specimen is sent to the pathologist who uses the imprint cytology technique to obtain a preliminary diagnosis. Effective counseling is established in case of breast cancer diagnosis. ResultsAmong 60 women, 25 patients were reassured by breast imaging and 35 underwent histopathological analysis (17 patients with a 1-day protocol and 18 with the standard definitive technique). Clinical examination was found to have a sensitivity of 100% and a specificity of 89.47%. The positive predictive value was 80 % and the negative predictive value was 100%. However, we did not find a strong correlation between imaging and definitive pathology. Moreover, on imprint cytology, sensitivity, specificity, PPV, and NPV were 100%. Finally, the mean time-to-treat was 28.6 days. ConclusionsSENODAY reassured 68.3% of patients. It also offered effective counseling and a treatment plan to newly diagnosed breast cancer patients within 1 day. Same day histological diagnosis by imprint cytology is effective and feasible with an excellent accuracy.

A Rapid, Sensitiveand Type-Specific Detection of High-Risk HPV-16 and HPV-18.

Human papillomavirus (HPV) infection, particularly infection with HPVs 16 and 18, is a major cause of cervical cancer. The current high-risk HPV screening or diagnosis tests use cytological or molecular techniques that are primarily based on qualitative HPV DNA detection. Comparative studies, however, revealed that different assays have varying sensitivities for detecting specific HPV types. Here, we developed and optimized a sensitive PCR (Polymerase Chain Reaction) assay for detection of high-risk HPV-16 and HPV-18. The PCR parameters were optimized, and analytical specificities were validated. Performance of developed PCR assay was evaluated in clinical samples (n = 100) which showed 100% specificity for both the assays and 96.97% and 94.12% sensitivity for HPV-16 and HPV-18, respectively. The developed assay demonstrated high sensitivity and specificity for detection of high-risk HPV-16 and HPV-18, making it applicable to routine HPV detection practices.

Relationship between a deep learning model and liquid-based cytological processing techniques.

Artificial intelligence (AI)-based cytopathology studies conducted using deep learning have enabled cell detection and classification. Liquid-based cytology (LBC) has facilitated the standardisation of specimen preparation; however, cytomorphology varies according to the LBC processing technique used. In this study, we elucidated the relationship between two LBC techniques and cell detection and classification using a deep learning model. Cytological specimens were prepared using the ThinPrep and SurePath methods. The accuracy of cell detection and cell classification was examined using the one- and five-cell models, which were trained with one and five cell types, respectively. When the same LBC processing techniques were used for the training and detection preparations, the cell detection and classification rates were high. The model trained on ThinPrep preparations was more accurate than that trained on SurePath. When the preparation types used for training and detection were different, the accuracy of cell detection and classification was significantly reduced (P < 0.01). The model trained on both ThinPrep and SurePath preparations exhibited slightly reduced cell detection and classification rates but was highly accurate. For the two LBC processing techniques, cytomorphology varied according to cell type; this difference affects the accuracy of cell detection and classification by deep learning. Therefore, for highly accurate cell detection and classification using AI, the same processing technique must be used for both training and detection. Our assessment also suggests that a deep learning model should be constructed using specimens prepared via a variety of processing techniques to construct a globally applicable AI model.

Development and Validation of a Simulation Model for Collection of Canine Vaginal Samples.

Vaginal cytology is a widely used cytological technique mostly taught by observation, either through direct tutoring or videos. To the best of our knowledge, vaginal cytology simulators have never been assessed in veterinary medicine. Twenty-five undergraduate students with no prior experience in canine vaginal sampling were randomly assigned to two groups that either practiced the procedure in a simulator or a live animal. An inverted classroom design was followed. After observing a video tutorial, students practiced with the simulator/live animal for two classes. Three weeks later, they performed a vaginal cytology on a live animal being recorded. The videos were evaluated through an objective structured clinical examination (OSCE) by an observer blinded to the student's groups. The learning outcome was compared through OSCE pass rates and questionnaires. The simulation model was made by 3D printing and soft silicone for the vulvar labia, having pink and blue colored vaseline in the correct and incorrect locations for sampling. The model was economic and accurately replicated the female reproductive tract. It provided immediate feedback to students, by obtaining pink or blue swabs from the correct and incorrect locations, respectively. Students reported that three to five or more attempts were needed to properly learn the procedure, thus justifying the need for a simulator. No differences in the OSCE pass rates were observed between the groups. The simulation model was effective for learning the vaginal cytology procedure, replacing the use of live animals. This low-cost model should be incorporated in the tool-kit of reproduction classes.

Deep Learning and Transfer Learning Methods to Effectively Diagnose Cervical Cancer from Liquid-Based Cytology Pap Smear Images

As cervical cancer is considered one of the leading causes of death for women globally, different screening techniques have emerged. As the Papanicolaou technique generates high numbers of false negatives due to only testing 20% of a sample, the liquid-based cytology technique was developed to test 100% of the sample and improve accuracy. However, as the larger sample size has made it difficult to detect the lesion images through a microscope, studies have looked for ways to intelligently analyze sample. The aim of this study is to develop an artificial intelligence image recognition system that detects the lesion level of cervical cancer of liquid-based Pap smears under the Bethesda classification of cancer (NI/LSIEL/HSIEL/SCC). For this purpose, six activities were carried out: dataset selection, data augmentation, optimization, model development, evaluation and system construction. A dataset built from publicly available Pap smear images and passed through data augmentation algorithms generated a total of 2,676 images. Two models, ResNet50V2 and ResNet101V2, were developed under Deep Learning and Transfer Learning protocols. The evaluation showed that the ResNet50V2 model obtained better performance, where the classification of HSIL and SCC type images obtained a precision of 0.98 and achieved an accuracy of 0.97. Finally, the system based on the ResNet50V2 model was built and its performance was validated.

CYTOLOGICAL CHARACTERIZATION OF TOMATO (SOLANUM LYCOPERSICUM L.) GERMPLASM

Germplasm diversity of any crop plays a significant role in the establishment of a successful breeding programme. In this study, five tomato germplasm, namely Roma Savannah, Tima, UTC, Dan Zaria and Tagino were collected from the Nasarawa State College of Agriculture, Lafia were evaluated for their cytological characterization to determine the germplasm cytological karyotyping parameters. The number of chromosomes in all lines was 2n = 24, except for Tagino, in which the number was 2n = 26, whereas chromosome sizes were small, ranging from 322.09 to 461.17µm. The Roma Savannah cultivar was a symmetrical cultivar (primitive), having the highest total form percentage (TF%) and symmetry index (Syi) values and the minimum karyotype asymmetry index (ASK) value, whereas Dan Zaria was asymmetrical (advanced) which makes it a superior germplasm among the other five evaluated. On the whole, the application of cytological techniques could be considered as a means to provide suitable parameters for studying the evolution of the genetic divergence between the studied tomato lines which can aid in preserving genetic resources in gene banks and improving new cultivars in breeding systems.

Role of Crush Cytology in Detecting Gastrointestinal Malignancies

Background: Adenocarcinomas are the most common malignancy of the gastrointestinal (GI) tract. Crush cytology is an effective method which can be used to detect neoplastic conditions of GI tract, especially in combination with biopsies. Materials and Methods: The objectives of the study were to study the efficacy of crush cytology as a convenient and near accurate method to evaluate endoscopic biopsy of GI neoplasms. Study Design: Retrospective and Prospective. The original cytopathology diagnoses were correlated with histology report on cases sent over 2-year period. In consecutive 89 patients attending the department of gastroenterology with clinical suspicion of malignancy, an endoscopy was performed. The material obtained was subjected to cytology as well as histopathology. The stained cytology slides along with corresponding histopathology slides were studied. Results: Out of 89 cases, both crush smear and histopathology sections were positive for malignant cells (65 cases), both negative for malignant cells (seven cases), crush smear positive and histopathology negative (three cases), and crush smear-negative and histopathology positive (14 cases). Out of the 14 cases which were negative in crush smears for malignancy, on histopathological examination, eight were signet-ring cell carcinoma, three were poorly differentiated adenocarcinoma, two turned out to be neuroendocrine carcinoma, and one non-Hodgkin lymphoma (NHL). Conclusion: Crush cytology smears can be used to diagnose malignant GI cases fairly accurately. Cases of signet-ring cell carcinoma, neuroendocrine tumor, poorly differentiated malignancy, and NHL may be missed by crush cytology technique alone. A special stain for mucin can be used to identify the signet-ring cells in signet-ring cell carcinoma cases.

Cellular diameter in desquamated cells of smokeless tobacco users

Background: Tobacco-induced mucosal changes have been identified in exfoliated cells. The morphology of the exfoliated cells depends on the nature of the changes taking place in the epithelial layer; conversely, alteration in cytological pattern may be attributed to the changes occurring in the epithelial layer. Applying this possibility, exfoliative cytological techniques have been applied to examine the effect of tobacco on the oral mucosa. Aims and Objectives: The study was conducted to study the alterations in the cellular diameter of buccal mucosal cells. Materials and Methods: In a case controlled study, 240 participants were divided into four groups according to their habit of consumption of tobacco leaves, tobacco with areca nut, participants with oral lesions, and participants without any such habit. The groups were, further, subdivided according to their age. Desquamated cells from the buccal mucosa of were collected and smeared on a slide. The cells were stained with Papanicolou (rapid kit). Fifty cells per slide were studied and photomicrographs were obtained at ×40 magnification. The cellular diameter was obtained using the Leica 1000 software. Results: The mean cell diameter was found to be highest in the group of normal subjects (49.51±7.31 micron), followed by the group of tobacco users (43.52±1.92 micron) and Gutkha users (39.05±3.57 micron). The mean cell diameter was minimum (24.97±3.13 micron) in the group of oral lesion patients. According to the ANOVA test, the difference in cell diameter among various groups is found to be highly significant (P&lt;0.0001). Conclusion: Tobacco used in any form can induce changes in the buccal mucosa. In our study, mean cellular diameter decreased from normal-cells to cells affected by oral lesions.

Nuclear and Nucleolar Morphometric Changes in MGP Stained Oral Epithelial Cells In Cigarette Smokers and Non Smokers

Aim: To compare nuclear and nucleolar morphometric changes in methyl green pyronin stained oral epithelial cells in cigarette smokers, passive smokers and non smokers Study design: A comparative cross sectional study Place and duration: Carried out in Postgraduate Medical Institute, Lahore. Duration was 6 months, March 2022 to August 2022. Methods: 120 subjects were equally divided into three main groups, cigarette smokers, passive smokers and non smokers, each fulfilling the inclusion and exclusion criteria. Buccal smears were collected by exfoliative cytology of mucosa and then stained by methyl green pyronin stain. The quantitative nuclear morphometric parameters i.e., nuclear and nucleolar diameter and area, and number of nucleoli, were measured. Results: Themedian of all nuclear parameters of smokers was significantly higher as compared to passive smokers and non-smokers in MGP stain (p ≤ 0.05). Further, effect of duration of smoking on morphometric variables showed significant (p&lt; 0.05) results amongst the smokers in MGP stain. However, no significant differences were seen in all morphometric parameters with respect to frequency of cigarette smoking. Conclusion: The study confirms nuclear morphometric changes in oral epithelial cells of cigarette smokers indicating that smoking does influence cellular alterations. The simplicity of cytology technique with the use of methyl green pyronin stain is valuable screening tool for detecting early oral mucosal changes in high risk patients. Keywords: smoking, exfoliative cytology, nuclear morphometric parameters, methyl green pyronin stain.

Development of a postoperative nomogram to predict overall survival in gastric cancer using variables the AJCC recommends for collection and registration.

468 Background: The American Joint Committee on Cancer (AJCC) has increasingly recognized the need for individual risk prediction model for the era of tailored therapy. This study aimed to develop a postoperative gastric cancer nomogram for prediction of overall survival (OS). Methods: The nomogram was developed using data of 4,990 patients with primary gastric cancer who underwent macroscopically complete resection (residual tumor: R0 or R1) at Shizuoka Cancer Center (Shizuoka, Japan), and it was created with a multivariable Cox proportional hazard regression model. Fifteen pathological or host-related variables (age, sex tumor location, tumor size, macroscopic type, histology, depth (pT), number of positive nodes (pN), number of negative nodes, location of positive nodes, lymphovascular invasion, lavage cytology (CY), tumor margin, serum CEA and serum CA19-9) were collected. They are recommended to collect and register by the AJCC. The model was validated internally using measures of discrimination (Harrell’s C-index), calibration and decision curve analysis. A stage-specific subset survival analysis of the three risk groups (low, intermediate, high) calculated using the nomogram was performed. Results: In the development procedure, multivariable analysis for OS selected 11 variables for constructing the nomogram. The developed nomogram showed good discrimination, with a C-index of 0.812; that of the American Joint Committee on Cancer (AJCC) pathological stage was 0.756. The nomogram performed well in the calibration and decision curve analyses. A stage-specific subset survival analysis of the three risk groups calculated using the nomogram also showed the superiority of nomogram-prediction when compared to AJCC ordinal staging. Conclusions: This new postoperative risk model accurately predicts OS in gastric cancer and can be used for patient counseling in clinical practice and clinical trial eligibility determination.