Artificial Intelligence in Toxicologic Pathology: Quantitative Evaluation of Compound-Induced Hepatocellular Hypertrophy in Rats

Digital pathology evolved rapidly, enabling more systematic usage of image analysis and development of artificial intelligence (AI) applications. Here, combined AI models were developed to evaluate hepatocellular hypertrophy in rat liver, using commercial AI-based software on hematoxylin and eosin-stained whole slide images. In a first approach, deep learning-based identification of critical tissue zones (centrilobular, midzonal, and periportal) enabled evaluation of region-specific cell size. Mean cytoplasmic area of hepatocytes was calculated via several sequential algorithms including segmentation in microanatomical structures (separation of sinusoids and vessels from hepatocytes), nuclear detection, and area measurements. An increase in mean cytoplasmic area could be shown in groups given phenobarbital, known to induce hepatocellular hypertrophy when compared to control groups, in multiple studies. Quantitative results correlated with the gold standard: observation and grading performed by board-certified veterinary pathologists, liver weights, and gene expression. Furthermore, as a second approach, we introduce for the first time deep learning-based direct detection of hepatocellular hypertrophy with similar results. Cell hypertrophy is challenging to pick up, particularly in milder cases. Additional evaluation of mean cytoplasmic area or direct detection of hypertrophy, combined with histopathological observations and liver weights, is expected to increase accuracy and repeatability of diagnoses and grading by pathologists.

Cytomorphometric Analysis of Buccal Exfoliated Cells in Patients With Iron Deficiency Anemia

Iron deficiency anemia can cause histopathologic alterations in the oral mucosa. Exfoliative cytology is a cheaper and less aggressive method for early diagnosis. The aim of this study was to compare the cytological and cytomorphometric characteristics of buccal exfoliated cells of iron deficiency anemic patients with those of healthy controls. This case-control study compared a group of 40 patients with iron deficiency anemia (IDA) with an age- and gender-matched control group (C) of 40 healthy individuals. The buccal mucosal smears were stained using the Papanicolaou technique for cytological analyses. Cellular clumping, cytoplasmic diameter (CD), nuclear diameter (ND), cytoplasmic area (CA), nuclear area (NA), nucleus to cytoplasmic area ratio (NA/CA), cellular and nuclear pleomorphism, micronuclei (Mn), binucleation, bacterial colonies, and keratin flakes were evaluated using a light microscope and digital image analysis. Mean values for IDA and C groups were: bacterial colonies (1.88 and 0.65; P=0.002); CA (2209.88 and 1687.79 μm²; P=0.006); Mn (1.60 and 0.60; P=0.02). Significant increases in bacterial colonies, CA and Mn were seen for the IDA group. The number of cellular clumps, CD, ND, NA, NA/CA, cellular and nuclear pleomorphism, binucleation, and the number of keratin flakes didn’t show significant differences between studied groups (P>0.05). There wasn’t any significant difference with respect to overall atypia. This study revealed that IDA was able to induce significant changes in CA and Mn of the oral epithelial cells. Exfoliative cytology and cytomorphometry can be used as a tool to assess the mucosal changes in IDA patients.

Effect of tobacco in human oral leukoplakia: a cytomorphometric analysis

Objectives. Tobacco use is one of the most critical risk factors for different oral diseases. The aim of this study is to demonstrate the effect of tobacco on oral mucosa by cytomorphometric analysis of cells with the help of exfoliative cytology and to find out the improvement in diagnostic sensitivity of exfoliative cytology in the detection of dysplastic changes and early oral malignancy. Methods. The nuclear area (NA) and cytoplasmic area (CA) of cells were measured within cytological smear obtained from leukoplakia lesions of buccal mucosa of 90 tobacco users, 30 smokers (TS), 30 chewers (TC) and 30 with combined habit of smoking and chewing (TSC)] and from normal buccal mucosa of 30 non users (NU) of tobacco. Each habit group consisted of 30 tobacco users with oral leukoplakia lesion with mild epithelial dysplasia only. The 30 non-users of tobacco served as controls. The mean values of the CA and NA were obtained for each case, and the nuclear/cytoplasmic area (NA/CA) ratio was calculated. Results. The results showed a statistically significant increase (P<0.001) in mean NA and a statistically significant decrease (P<0.001) in mean CA values of tobacco users with leukoplakia as compared to non-users, hence NA/CA ratio value was significantly higher in tobacco users with the lesion. Conclusion. The changes in cellular morphology caused by tobacco use can be visualized by use of exfoliative cytology with the help of cytomorphometric analysis. The evaluation of parameters (NA, CA and NA/CA ratio) may increase the sensitivity of exfoliative cytology for the early diagnosis of oral premalignant and malignant lesions.

ANALYSIS OF SULFUR VAPOR EXPOSURE TO THE NUMBER OF MICRONUCLEUS AND ORAL BUCCAL EPITHELIAL MORPHOLOGY

Background: The sulfur vapor consists of SO2 and CO2 which are genotoxins that may cause the damage of DNA to the micronucleus in buccal epithelial cells. Micronucleus is a mass like a nucleus, measuring one-third of the nucleus. DNA damage can also be seen from changes in the morphology of epithelial cells. Objective: This study aimed to identify the effect of sulfur vapor exposure on the number of micronucleus and morphology epithelial cells in the oral cavity on the sulfur miner. Methods: The method of this study was analytic observational with a cross-sectional approach. The total sample of this study was 24 respondents divided into 2 groups, each group contained 12 respondents. Exfoliated buccal cells were collected by scrapping the buccal mucosa. The specimens stained using Hematoxylin and Eosin. Nucleus and cytoplasmic area were examined using image J 1.40 Results: The result showed the average number of buccal mucosa micronucleus on coal miners higher (35,50) than controls (11,58). The result of Independent-measures T-test obtained significant different on the number of micronucleus between sulfur miner and controls (p=0,000). The result of Independent-measures T-test on the nuclear area and cytoplasmic area between sulfur miner and controls obtained insignificant different (p=0,379 dan p=0,616). Conclusion: Based on this study can be concluded that sulfur vapor exposure affected on the number of micronucleus on sulfur miners, but did not influence morphology of epithelial cells.

A cytomorphometric analysis of the oral mucosa in patients with type 2 diabetes mellitus

Background: Although many of the pathological conditions of oral mucosa are clinically distinguishable, most lesions require a definitive diagnosis. This article tried the use of exfoliative cytology as an alternative tool in the screening, diagnosis and follow-up of diabetes mellitus.Materials and Methods: After rinsing the mouth with normal saline, slides were prepared from buccal mucosa and dorsum of tongue and fixed in 95% ethyl alcohol. The slides were stained with Papanicolaou stain and Acridine orange. Fifty clearly defined cells in each slide were visualized under light microscope for cytomorphometric analysis of cells using Image J software and under fluorescence microscope for assessment of nuclear alterations like micronuclei, nuclear budding, binucleation, multinucleation and karyorrhexis.Results: Statistically significant increase in Nuclear area BM (p = 0.000057), Nuclear Area Tongue (p= 0.0000113), total Nuclear Area (p= 000079), Cellular Area BM (p= 0.0475), Cellular Area Tongue (p= 0.0105), Total Cellular Area (p= 0.00496), Cytoplasmic Area Tongue (p= 0.00358), Total Cytoplasmic Area (p= 0.00268) were obtained from epithelial cells in the diabetic group when compared with the control group. Also the epithelial cells from the diabetic group showed features such as nuclear budding, micronuclei, binucleation, karyorrhexis and perinuclear halo. Conclusion: The objective demonstration of cytomorphometric and nuclear alterations by the oral exfoliated cells indicate the presence of cytological changes in the oral mucosa of diabetic patients despite the apparently normal clinical appearance. Hence, cytomorphometric analysis would aid the health professional as an additional non-invasive tool for the screening and monitoring of Diabetes Mellitus.

Morphometric Characterization of Small Cell Lymphocytic Lymphoma

The morphometry in histopathology is used to characterize cell populations belonging to different tissues and to identify differences in their parameters with prognostic implications. To achieve morphometric examination were selected 6 of 24 cases identified as small cell lymphocytic lymphoma. For each case analysis was done on five fields, for each field measuring the parameters of 20 cells. The studied parameters were for cytoplasm: cytoplasmic area, maximum and minimum cytoplasmic diameter, cytoplasmic perimeter; for nucleus were measured: nuclear area, minimum and maximum nuclear diameter, nuclear perimeter, nuclear contour index, nuclear ellipticity index, nuclear irregularity index. Also the nucleocytoplasmic ratio was calculated in all studied cases. Small cell lymphocytic lymphoma is characterized in morphometric terms having a small cytoplasmic area (average 29.206) and also a small nuclear area (mean 28.939) having a nucleo-cytoplasmic ratio appearance suggestive for adult lymphocyte. A nuclear contour index small value (3.946), ellipticity index value also small (3.521) and small nuclear irregularity index (3.965). Standard deviations, in any of the studied morphometric categories, is around or below 1 suggesting monomorphic cell appearance. These morphometric and microscopic features characterized mainly by a small population of adult lymphocytes, monomorphic, with rounded hipercromic nuclei, dense chromatin, support the framing into indolent lymphoma group in terms of clinical outcome.

Cytological analysis of epithelial cells in adolescents with type 1 diabetes mellitus

Introduction: Type 1 diabetes mellitus is a chronic metabolic disorder condition, which may causes oral mucosa changes in adolescents. The effect of glycemic control on the oral tissues has been scarcely reported as well as the low number of oral mucosal lesions found in this population. The goal of this study is to evaluate cytopathological changes in oral epithelial cells of adolescents with type 1 diabetes mellitus (DM1) and assess the glycemic control. Materials and methods: Oral smears were collected from normal mucosa by exfoliative cytology in 40 healthy adolescents and 40 with DM1. Cell morphology and cellularity were analyzed. Nuclear area (NA) and cytoplasmic area (CA) were measured; nuclear/cytoplasmic area ratio (NA/CA) was calculated. Time elapsed from the diagnosis of diabetes and glycated hemoglobin levels (HbA1c) were recorded. Results: There was no significant difference to NA, CA, NA/CA for both groups and also in relation to HbA1c (P > 0.05). No morphological differences were found among the groups (P > 0.05). There was a predominance of nucleated cells of the superficial layer in smears of both groups. Class I and Class II smears were predominant in both groups. Conclusion: This study revealed that type 1 diabetes mellitus and the glycemic control were unable to induce significant changes on oral epithelial cells in adolescents.

Myenteric neuronal plasticity induced by Toxoplasma gondii (genotype III) on the duodenum of rats

The effects of acute and chronic infection caused by Toxoplasma gondii on duodenal myenteric neurons were analyzed. Eighteen rats were assigned into four groups: Acute Control Group (ACG, n=4); Acute Experimental Group (AEG, n=4); Chronic Control Group (CCG, n=5); and Chronic Experimental Group (CEG, n=5). Rats from the AEG and CEG were inoculated orally with 105 genotype III (BTU-II strain) tachyzoites of T. gondii isolated from a dog with neurological signs. Acute groups were killed after 24 hours after the inoculation and the chronic groups after 30 days. Whole-mount from the duodenum were stained with Giemsa. The population density of myenteric neurons, as well the body cell, nuclear and cytoplasmic area were analyzed. Both acute and chronic toxoplasmic infection did not provoke neuronal loss. On the other hand, plastic alterations were observed: decreasing of the nuclear and cytoplasmic area during the acute phase and neuronal hypertrophy during the chronic phase.

Effects of Humoral Hypercalcemia of Malignancy on the Parathyroid Gland in Nude Mice

The ultrastructure of parathyroid chief cells was examined from four groups of nude mice (NIH: Swiss) with different serum calcium concentrations. The groups consisted of eight male mice with hypercalcemia induced by transplantable canine adenocarcinoma (CAC-8), eight female mice with hypercalcemia induced by infusion of parathyroid hormone-related protein, ten male control mice, and six male mice fed a low calcium (0.01%) diet. Hypercalcemia induced by malignancy or parathyroid hormone-related protein infusion was associated with low serum phosphorus concentration, a decrease in the number of secretory and prosecretory granules in the parathyroid chief cells, and an increase in the cytoplasmic area of chief cells. Prominent myelinlike membranous whorls were present in the cytoplasm of chief cells of tumor-bearing and parathyroid hormone-related protein-infused hypercalcemic mice. Mice fed a low calcium diet had decreases in the number of secretory granules and cell area but increases in the number of prosecretory granules compared with control mice. The number of mitochondria and the nuclear area of chief cells were similar in all four groups. The prominent membranous whorls and increased cytoplasmic area of chief cells from these hypercalcemic mice mark these cells as distinctly different from the parathyroid chief cells of other species with hypercalcemia.