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Animals : An Open Access Journal From... May 2023The liver is the main site of fat synthesis and plays an important role in the study of fat deposition in poultry. In this study, we investigated the developmental...
The liver is the main site of fat synthesis and plays an important role in the study of fat deposition in poultry. In this study, we investigated the developmental changes of duckling livers and isolated primary duck hepatocytes. Firstly, we observed morphological changes in duckling livers from the embryonic period to the first week after hatching. Liver weight increased with age. Hematoxylin-eosin and Oil Red O staining analyses showed that hepatic lipids increased gradually during the embryonic period and declined post-hatching. Liver samples were collected from 21-day-old duck embryos for hepatocyte isolation. The hepatocytes showed limited self-renewal and proliferative ability and were maintained in culture for up to 7 days. Typical parenchymal morphology, with a characteristic polygonal shape, appeared after two days of culture. Periodic acid-Schiff (PAS) staining analysis confirmed the characteristics of duck embryo hepatocytes. PCR analysis showed that these cells from duck embryos expressed the liver cell markers ALB and CD36. Immunohistochemical staining and immunofluorescence analysis also confirmed ALB and CK18 expression. Our findings provide a novel insight regarding in vitro cell culture and the characteristics of hepatocytes from avian species, which could enable further studies concerning specific research on duck lipid metabolism.
PubMed: 37889689
DOI: 10.3390/ani13111820 -
Sultan Qaboos University Medical Journal Feb 2021The post-mortem interval (PMI) refers to the amount of time elapsed between death and discovery of the body. This study aimed to evaluate light microscopic cellular...
OBJECTIVES
The post-mortem interval (PMI) refers to the amount of time elapsed between death and discovery of the body. This study aimed to evaluate light microscopic cellular changes in the oral mucosa and identify the potential of this method for predicting PMI.
METHODS
This prospective study was conducted between July 2016 and January 2018 at the Institute of Dental Sciences, Siksha 'O' Anusandhan University, Bhubaneswar, India. A total of 150 post-mortem (including 75 gingival and 75 buccal mucosa samples) and 40 ante-mortem (including 20 gingival and 20 buccal mucosa samples) tissue samples were compared using haematoxylin and eosin, periodic acid-Schiff (PAS) and van Gieson stains. Microscopic changes in the epithelium and connective tissue were categorised according to PMI stage as early (<12.5 hours since death), intermediate (12.5-20.5 hours since death) or late (>20.5 hours since death).
RESULTS
Most epithelial cellular changes occurred early, except for arc-shaped nuclei and epithelial shredding which were intermediate and late changes, respectively. However, microscopic changes in the connective tissue were only observable at ≥12.5 hours. There was a progressive decrease in intensity in van Gieson stains and an increase in intensity in PAS stains as PMI increased. Several microscopic features were found to be significant predictors of PMI including epithelial homogenisation, cytoplasmic vacuolation, nuclear degeneration, arc-shaped nuclei, chromatin clumping, red blood cell clumping and lysis, melanin incontinency, myofibril degeneration, salivary gland degeneration and epithelial connective tissue separation ( <0.050 each).
CONCLUSION
These findings indicate that microscopic evaluation of the oral mucosa may be helpful for PMI prediction.
Topics: Histocytochemistry; Humans; India; Mouth Mucosa; Postmortem Changes; Prospective Studies; Salivary Glands
PubMed: 33777421
DOI: 10.18295/squmj.2021.21.01.005 -
Journal of the American Society of... Jan 2021Nephropathologic analyses provide important outcomes-related data in experiments with the animal models that are essential for understanding kidney disease...
BACKGROUND
Nephropathologic analyses provide important outcomes-related data in experiments with the animal models that are essential for understanding kidney disease pathophysiology. Precision medicine increases the demand for quantitative, unbiased, reproducible, and efficient histopathologic analyses, which will require novel high-throughput tools. A deep learning technique, the convolutional neural network, is increasingly applied in pathology because of its high performance in tasks like histology segmentation.
METHODS
We investigated use of a convolutional neural network architecture for accurate segmentation of periodic acid-Schiff-stained kidney tissue from healthy mice and five murine disease models and from other species used in preclinical research. We trained the convolutional neural network to segment six major renal structures: glomerular tuft, glomerulus including Bowman's capsule, tubules, arteries, arterial lumina, and veins. To achieve high accuracy, we performed a large number of expert-based annotations, 72,722 in total.
RESULTS
Multiclass segmentation performance was very high in all disease models. The convolutional neural network allowed high-throughput and large-scale, quantitative and comparative analyses of various models. In disease models, computational feature extraction revealed interstitial expansion, tubular dilation and atrophy, and glomerular size variability. Validation showed a high correlation of findings with current standard morphometric analysis. The convolutional neural network also showed high performance in other species used in research-including rats, pigs, bears, and marmosets-as well as in humans, providing a translational bridge between preclinical and clinical studies.
CONCLUSIONS
We developed a deep learning algorithm for accurate multiclass segmentation of digital whole-slide images of periodic acid-Schiff-stained kidneys from various species and renal disease models. This enables reproducible quantitative histopathologic analyses in preclinical models that also might be applicable to clinical studies.
Topics: Algorithms; Animals; Deep Learning; Diagnosis, Computer-Assisted; Disease Models, Animal; Image Processing, Computer-Assisted; Kidney; Kidney Diseases; Kidney Glomerulus; Male; Mice; Mice, Inbred C57BL; Neural Networks, Computer; Pattern Recognition, Automated; Periodic Acid; Reproducibility of Results; Schiff Bases; Translational Research, Biomedical
PubMed: 33154175
DOI: 10.1681/ASN.2020050597 -
Animals : An Open Access Journal From... May 2023In terrestrial mammals, the parotid and mandibular glands secrete different types of saliva into the oral cavity. Both glands were obtained from two female lowland...
In terrestrial mammals, the parotid and mandibular glands secrete different types of saliva into the oral cavity. Both glands were obtained from two female lowland tapirs () and one female aardvark () from the Wroclaw Zoological Garden (Poland) and examined by light microscopy (hematoxylin and eosin, mucicarmine, periodic acid-Schiff, Alcian blue pH 1.0, Alcian blue pH 2.5, Alcian blue pH 2.5/PAS, and Hale's dialysed iron). Both the parotid glands observed in the lowland tapir and aardvark were compound alveolar serous secretory units, and in both species, the secretion was composed of neutral and acidic mucopolysaccharides (sialo and sulfated mucins). However, in both the lowland tapir and aardvark, a histological examination found the stroma of the mandibular gland was divided into very large lobes by poorly marked connective tissue. While many interlobar and striated ducts were found in the aardvark, very few were found in the lowland tapir. The mandibular gland was a branched tubular (mucous secretion) type in the lowland tapir, but it was a branched tubuloalveolar (mucous-serous) type in the aardvark. In all tested glands, the secretion was composed of neutral mucopolysaccharides, acid-sulfated mucosubstances, and sialomucins.
PubMed: 37238113
DOI: 10.3390/ani13101684 -
Cureus Nov 2022Introduction Chronic calculous cholecystitis is the most common and important contributing factor for cholecystectomy across the country, with a prevalence of 2-29%....
Quantitative Analysis of Mucin Expression Using Combined Alcian Blue-Periodic Acid Schiff (AB-PAS) Stain and Combined High Iron Diamine-Alcian Blue (HID-AB) Stain and the Correlation With Histomorphological Score in Chronic Calculous Cholecystitis.
Introduction Chronic calculous cholecystitis is the most common and important contributing factor for cholecystectomy across the country, with a prevalence of 2-29%. Cholesterol supersaturated bile plays a major role in stone formation. It is very essential to identify the pathogenesis of stone formation in order to prevent its formation. This study is aimed to evaluate histomorphological features of chronic calculous cholecystitis and to quantitatively evaluate alteration in mucin expression using the combined Alcian blue-periodic acid Schiff (AB-PAS) stain and the combined high iron diamine Alcian blue (HID-AB) stain, to correlate with each other and also with biochemical features of gall stones. Methods A cross-sectional study of 64 chronic calculous cholecystitis were taken for histomorphological assessment and grading using Hematoxylin and Eosin (H&E) stain and Masson's trichrome stains. Expression of the type of mucins was analyzed using histochemical stains by a standardized scoring system. Results A significant positive correlation was observed between an increase in grades of inflammation and fibrosis with an increase in the quantity of sialomucin and neutral mucin in the deep layers of epithelium, and a significant negative correlation was observed between an increase in grades with a decrease in acidic mucin and sulfomucin of both superficial and deep epithelium except sulfomucin in fibrosis. No significant correlation was obtained with muscle thickness, adipose tissue deposition, and epithelial hyperplasia. A higher frequency of mixed-type stones was associated with severe inflammation. Conclusion Inflammation and fibrosis were strongly correlated with quantitative alteration and reversal of mucin composition in chronic cholecystitis; hence we conclude that these features play a significant role in the pathogenesis of stone formation. Using Combined AB(2.5pH)-PAS stain and Combined HID-AB(2.5 pH) stain to detect mucin hypersecretion and composition of altered mucin is relatively accurate and cost-effective rather than performing costly immunohistochemical (IHC) markers.
PubMed: 36600870
DOI: 10.7759/cureus.32033 -
Romanian Journal of Morphology and... 2022Maternal obesity is associated with increased maternal and fetal morbidity and mortality, with an increased risk of gestational diabetes mellitus (GDM) and preeclampsia...
Maternal obesity is associated with increased maternal and fetal morbidity and mortality, with an increased risk of gestational diabetes mellitus (GDM) and preeclampsia (PE). This prospective study histopathologically analyzes the placentas obtained from 34 pregnant obese women studied between October 2016 and May 2020. The 10 cases of term placentas from obese pregnancies with GDM and the 12 cases with PE were examined by the Hematoxylin-Eosin (HE), Masson's trichrome (MT) and Periodic Acid-Schiff-Hematoxylin (PAS-H) classical stainings, and by the immunohistochemical evaluation and compared to placentae from uncomplicated term obese pregnancies (12 cases). We did not meet placental histopathological (HP) abnormalities that we could classify as characteristic only for the state of obese pregnancy, but we did find placental changes associated with PE and GDM, in the context of obese pregnancy. In the case of association with PE, there were common lesions, manifested by intra- and perivillous fibrinoid deposition, calcification, and placental infarction area, to which were added numerous syncytial knots. In the case of obese pregnancy associated with GDM, we found, in addition to common placental lesions of obesity, intravillositary vascular edema and in the terminal villi appearing chorangiosis. This study revealed a number of HP changes that occur in maternal obesity, even in uncomplicated obese pregnancies. A characteristic of obese pregnancies associated with PE was the presence of numerous syncytial knots, and in obese pregnancies associated with GDM, the most common HP lesion was placental chorangiosis. Certainly, we cannot conclude that these HP lesions are specific to a particular pathology, but they belong primarily to the status of maternal obesity.
Topics: Diabetes, Gestational; Female; Hematoxylin; Humans; Obesity; Obesity, Maternal; Placenta; Pre-Eclampsia; Pregnancy; Pregnancy Outcome; Prospective Studies
PubMed: 36074672
DOI: 10.47162/RJME.63.1.09 -
The Malaysian Journal of Medical... Oct 2020This study was undertaken to evaluate the preferred method (Giemsa or periodic acid Schiff-Alcian blue [PAS-AB] stains) of detecting () in gastric mucosal biopsies in...
Evaluation of Better Staining Method among Hematoxylin and Eosin, Giemsa and Periodic Acid Schiff-Alcian Blue for the Detection of Helicobacter pylori in Gastric Biopsies.
BACKGROUND
This study was undertaken to evaluate the preferred method (Giemsa or periodic acid Schiff-Alcian blue [PAS-AB] stains) of detecting () in gastric mucosal biopsies in terms of sensitivity, specificity and applicability. To the best of my knowledge, this is the first report comparing Giemsa and PAS-AB staining for the detection of in such biopsies.
METHODS
The formalin-fixed paraffin-embedded blocks of 49 gastric biopsies from different patients were collected from the archive of anatomical pathology at King Abdulaziz Medical City, National Guard, Riyadh, Saudi Arabia. From each block, three slides were prepared and analysed using the hematoxylin and eosin (H&E), Giemsa and PAS-AB stains to detect the presence/absence of , and the results were compared in terms of sensitivity, specificity and applicability.
RESULTS
The majority of the biopsies in this study showed antrum-type gastric mucosa. Only 15 biopsies showed active gastritis, whereas the rest showed chronic gastritis. Three biopsies showed intestinal metaplasia. All were detected by PAS-AB stain, but only two-thirds were detected by H&E stain. Fifteen gastric biopsies showed infection in general and in 13 of them, active gastritis cases were discovered. Fourteen out of these 15 infection cases were detected by Giemsa stain, whereas only 13 cases were detected by H&E stain. PAS-AB stain showed the worst results since it demonstrated only 40% sensitivity and 67.65% specificity in detection.
CONCLUSION
Giemsa stain has better sensitivity and specificity in gastric infection detection than PAS-AB. Therefore, using PAS-AB stain to detect infection is not recommended.
PubMed: 33154702
DOI: 10.21315/mjms2020.27.5.6 -
Molecules (Basel, Switzerland) Oct 2023As a common emerging environmental pollutant, microplastics (MPs) have been detected in a variety of environmental media and human bodies. The potential toxic effects...
As a common emerging environmental pollutant, microplastics (MPs) have been detected in a variety of environmental media and human bodies. The potential toxic effects and mechanisms of MPs need to be revealed urgently. MPs can be deposited in the kidney, and exposure to high doses of MPs can cause nephrotoxicity in experimental animals. In this study, we investigated the effects of exposure to polystyrene microplastics (PS-MPs) at environmentally relevant doses (0.1 and 1 mg/L) on kidney structure, function, and transcriptome in mice. We found that mice exposed to PS-MPs in drinking water for eight weeks had no change in body weight or kidney coefficient. PS-MPs administration decreased the levels of blood urea nitrogen (BUN) in mice, while serum creatinine (CRE) and uric acid (UA) concentrations were unaffected. Through using periodic acid-Schiff (PAS) and Masson staining, we discovered that the glomerular tuft area increased in the PS-MP-treated mice, while the degree of renal fibrosis remained unchanged. Furthermore, renal cortex transcriptomic analysis identified 388 and 303 differentially expressed genes (DEGs) in the 0.1 and 1 mg/L dose groups, respectively. The DEGs were highly enriched in mitochondrial-related terms and pathways of thermogenesis and oxidative phosphorylation. Moreover, protein-protein interaction (PPI) network analysis revealed that cytochrome b-c1 complex subunit 10 (UQCR11) and cytochrome c oxidase subunit 3 (MT-CO3) were important node proteins. These findings suggest that environmental exposure to MPs can cause abnormalities in renal structure and filtration function and that long-term exposure to MPs may be a risk factor for renal disease.
Topics: Humans; Animals; Mice; Plastics; Transcriptome; Microplastics; Kidney; Kidney Glomerulus; Polystyrenes; Water Pollutants, Chemical
PubMed: 37894583
DOI: 10.3390/molecules28207104 -
Microorganisms Jun 2022In the wake of the COVID-19 pandemic, fungal infections of the maxillofacial region have become prevalent, making their accurate diagnosis vital. Histopathological...
In the wake of the COVID-19 pandemic, fungal infections of the maxillofacial region have become prevalent, making their accurate diagnosis vital. Histopathological staining remains a simple, cost-effective technique for differentiation and diagnosis of the causative fungal organisms. The present study aims to evaluate the staining efficacy of Periodic Acid-Schiff (PAS), Alcian Blue, Safranin-O and Gomori's Methenamine Silver (GMS) on fungal smears. This research work also attempts to study the morphometric characteristics of , , . , and , 10 smears each, were stained using PAS, Alcian Blue, Safranin-O and GMS. The morphological characteristics and staining efficacy were examined, and semi-quantitative scoring was performed. , and were stained for the first time with Safranin-O. The morphometric traits were then analyzed using an image analysis software. Safranin-O provided the most reliable staining efficacy amongst the stains and optimum morphological definition for all three organisms. Safranin-O was found to be superior to PAS and GMS, ensuring detection of even the most minute mycotic colonies. The hyphae of to be the largest, and the spores and fruiting body of were found to be the largest amongst the three organisms compared. Early and accurate diagnosis of fungal infections can significantly reduce morbidity in orofacial fungal infections.
PubMed: 35744745
DOI: 10.3390/microorganisms10061226 -
Evidence-based Complementary and... 2022The mitochondrial dysfunction of HCE-2 human corneal epithelial cells was induced by high osmotic pressure exposure and treated with resveratrol (50 M). Western...
METHODS
The mitochondrial dysfunction of HCE-2 human corneal epithelial cells was induced by high osmotic pressure exposure and treated with resveratrol (50 M). Western blotting was used to detect the expression of the antioxidant proteins SOD2, GPx, and SIRT1, and flow cytometry was used to detect cell apoptosis and ROS production. The DED mouse model was induced by 0.2% benzalkonium chloride (BAC) and treated with resveratrol. The tear yield was measured by the phenol cotton thread test, the density of cup cells in the conjunctiva was measured by periodic acid-Schiff (PAS) staining, and the expression levels of SIRT1, GPx, and SOD2 in lacrimal glands were detected by Western blotting.
RESULTS
In hypertonic conditions, the apoptosis of HCE-2 cells increased, the expression of the antioxidant proteins SOD2 and GPx decreased, ROS production increased, and the expression of SIRT1 protein, an essential regulator of mitochondrial function, was downregulated. Treatment with resveratrol reversed the mitochondrial dysfunction mediated by high osmotic pressure. In the DED mouse model, resveratrol treatment promoted tear production and goblet cell number in DED mice, decreased corneal fluorescein staining, upregulated SIRT1 expression, and induced SOD2 and GPx expression in DED mice.
CONCLUSION
Resveratrol alleviates mitochondrial dysfunction by promoting SIRT1 expression, thus reducing ocular surface injury in mice with dry eye. This study suggests a new path against DED.
PubMed: 35664941
DOI: 10.1155/2022/1013444