Counting ovarian early follicles helps assess the impact of pharmaceuticals and endocrine-disrupting chemicals on female fertility. Traditional manual counting methods are labor-intensive, time-consuming, and prone to intra- and interobserver variability, leading to inconsistent results. This study addresses these limitations by developing a fully automated solution for the identification and quantification of early-stage follicles in hematoxylin and eosin-stained rat ovarian sections, with the aim of enhancing efficiency, accuracy, and reproducibility. We developed a deep learning model designed to identify primordial type 1 and type 2 follicles, and a second model focusing on detecting early growing follicles. The automated system exhibited high performance on an independent test data set of 200 whole-slide images, whose predictions were validated by pathologists. The models achieved high precision and recall, with 98.49% and 90.92%, respectively, for type 1; 95.97% and 97.54% for type 2; and 99.45% and 80.09% for early growing follicles. Further comparisons with manual annotations performed by 4 independent pathologists revealed significant interobserver variability. While human performance was hindered by factors such as fatigue and inconsistent criteria application, resulting in variable precision and recall and high rates of missed follicles, the algorithm maintained superior consistency and overall higher F1-scores. Furthermore, a comparison between manual work and the algorithm’s productivity showed that the algorithm was 11 times faster than manual quantification. These results demonstrate that the automated approach not only matches expert accuracy but also effectively eliminates subjectivity and diagnostic drift inherent in manual counting, providing a reproducible method for follicle counting, and significantly enhancing workflow efficiency.
Galectins play an important role in intestinal immune modulation. Little is known about galectin expression in the normal gastrointestinal tract (GIT) of dogs or their potential role in canine immunosuppressant-responsive (IR) chronic inflammatory enteropathy (CIE). In this study, we described and compared the expression of galectins in the stomach, duodenum, ileum, and colon of dogs without CIE-IR (healthy dogs) ( n = 5) and dogs with CIE-IR ( n = 7). Cellular localization of galectins was evaluated using immunohistochemistry quantified with open-source digital pathology software (QuPath). Immunoreactivity in the normal GIT was intense for Gal-2, weak for Gal-3, and moderate for Gal-4. Gal-2 was expressed throughout the gastric mucosa, including epithelium, pits, glands, and lamina propria, whereas Gal-3 and Gal-4 were expressed in the lower two-thirds of the mucosa. In healthy dogs, inflammatory cells, myenteric plexus neurons, and smooth muscle cells were immunoreactive for the examined galectins, whereas enteroendocrine and goblet cells were only immunoreactive for Gal-4. Quantitatively, Gal-2 expression was significantly higher than Gal-3 and Gal-4 in the stomach and duodenum ( P < .05) and higher than Gal-3 in the ileum. In the colon, Gal-2 and Gal-4 expressions were significantly ( P < .05) higher than Gal-3. The expressions of Gal-2, Gal-3, and Gal-4 in duodenum, ileum, and colon of dogs with CIE-IR were significantly ( P < .05) lower than in healthy dogs. A potential role for Gal-2, Gal-3, and Gal-4 in GIT immune regulation and their involvement in the pathogenesis of CIE-IR in dogs is possible and warrants further study.
Dehydropyrrolizidine alkaloid (PA) poisoning is common in livestock, wildlife, and humans. Dehydropyrrolizidine alkaloids have varying toxicities, though comparisons characterizing individual alkaloid toxicities are incomplete. The objective of this work was to directly compare the toxicity of select PAs in male C57BL/6J mice. Mice were exposed to varying doses of riddelliine, senecionine, seneciphylline, lasiocarpine, heliotrine, riddelliine N-oxide, and senecionine N-oxide for 10 days. Hepatic necrosis, serum biochemistries, and hepatic pyrrole concentrations were compared. Riddelliine and senecionine free base and N-oxide, seneciphylline, and lasiocarpine caused hepatocellular degeneration and necrosis. Both senecionine alkaloids produced midzonal or periportal lesions. Similar, but less severe midzonal degeneration and necrosis were observed in high-dose seneciphylline-treated mice with minimal hepatocyte enlargement at low doses. Both riddelliine alkaloids induced centrilobular degeneration and necrosis. High-dose lasiocarpine caused periportal hepatocyte enlargement and necrosis. High-dose heliotrine did not cause necrosis. Hepatic necrosis scores, liver enzyme activities, and liver pyrrole concentrations correlated with dose and necrosis scores for all alkaloids except heliotrine and lasiocarpine. The affected lobular area, degree of hepatic degeneration and necrosis, and the production of pyrroles were PA-specific. Under these conditions, seneciphylline and senecionine alkaloids were most toxic followed by riddelliine alkaloids. Lasiocarpine and heliotrine were the least toxic. This differs from relative toxicities reported in other models. In conclusion, in these mice, there are unique individual PA toxicities, lesion distributions, and pyrrole concentrations.
