Periodic acid-Schiff Staining Methods Research Articles

A Periodic Comparison of Harderian Gland in Henna Partridge (Alectoris chukar) According to Different Developmental Stages

Background: The present study aimed at determining histological structure and histochemical properties of the henna partridge (Alectoris chukar) Harderian gland periodically. Objectives: For this purpose, the Harderian glands of 12 (6 females + 6 males) healthy henna partridges aged 3 and 6 months were used. Methods: Totally removed Harderian gland tissue samples were kept in 10% neutral formolin for 36 hours and fixed. After washing, dehydrating, and polishing with known histological techniques, tissue pieces were blocked in paraffin. Results: We found that a thin connective tissue capsule surrounds the Harderian gland of the henna partridge and that the capsule sends septums into the gland and divides the gland into lobes and lobules. It was observed that the corpus glandulae consist of low or high prismatic epithelial cells. Within each lobe, there is a single main draining channel which has a very large lumen along with primary draining channels. Primary and main draining channel epithelial cells have a single-layered cuboidal structure. By Gordon-Sweet staining, it was found that reticular fiber bundles started from the connective tissue capsule that surrounded the organ and spread by branching. The reticular fiber bundles extended in thin strands to the periphery of the corpus glandulae. These reticular fiber bundles in the regions where the connective tissue septums widened are thicker and form a network. The reticular fiber bundles are seen in the basal part of the epithelial cells forming the main draining channel and especially in the crypt areas. Reticular fibers are also found around blood vessels. Strong Alcian blue (AB)-positive epithelial cells are observed in the corpus glandulae in AB pH=2.5 staining, but no periodic acid-Schiff (PAS)-positive epithelial cells are observed in PAS staining. While the majority of primary draining channel cells show a strong AB-positive reaction, a few cells showed a weak PAS-positive reaction. Regarding the main draining channel epithelium, goblet cells show a weak and strong AB-positive reaction in AB pH=2.5 staining method, respectively and a weak PAS-positive reaction in PAS staining method. Conclusion: In line with these findings, no significant difference was found in terms of general histological structure and histochemical properties of the 3- and 6-month-old henna partridge Harderian gland.

Ticlopidine induces embryonic development toxicity and hepatotoxicity in zebrafish by upregulating the oxidative stress signaling pathway

Ticlopidine exerts its anti-platelet effects mainly by antagonizing platelet p2y12 receptors. Previously, a few studies have shown that ticlopidine can induce liver injury, but the exact mechanism of hepatotoxicity remains unclear. Oxidative stress, metabolic disorders, hepatocyte apoptosis, lipid peroxidation, and inflammatory responses can all lead to hepatic liver damage, which can cause hepatotoxicity. In this study, in order to deeply explore the potential molecular mechanisms of ticlopidine -induced hepatotoxicity, we used zebrafish as a model organism to comprehensively evaluate the hepatotoxicity of ticlopidine and its associated mechanism. Three days post-fertilization, zebrafish larvae were exposed to varying concentrations (1.5, 1.75 and 2 μg/mL) of ticlopidine for 72 h, in contrast, adult zebrafish were exposed exposure to 4 μg/mL of ticlopidine for 28 days. Ticlopidine-exposed zebrafish larvae showed changes in liver morphology, shortened body length, and delayed development of the swim bladder development. Liver tissues of ticlopidine-exposed zebrafish larvae and adults stained with Hematoxylin & Eosin revealed vacuolization and increased cellular interstitial spaces in liver tissues. Furthermore, using Oil Red O and periodic acid-Schiff staining methods and evaluating different metabolic enzymes of ticlopidine-exposed zebrafish larvae and adults suggested abnormal liver metabolism and liver injury in both ticlopidine-exposed zebrafish larvae and adults. Ticlopidine also significantly elevated inflammation and oxidative stress and reduced hepatocyte proliferation. During the rescue intervention using N-acetylcysteine, we observed significant improvement in ticlopidine-induced morphological changes in the liver, shortened body length, delayed swim bladder development, and proliferation of liver tissues showed significant improvement. In conclusion, ticlopidine might inhibit normal development and liver proliferation in zebrafish by upregulation of oxidative stress levels, thus leading to embryonic developmental toxicity and hepatotoxicity. In this study, we used zebrafish as a model organism to elucidate the developmental toxicity and hepatotoxicity induced by ticlopidine upregulation of oxidative stress signaling pathway in zebrafish, providing a theoretical basis for clinical application.

Severe choline deficiency induces alternative splicing aberrance in optimized duck primary hepatocyte cultures.

Choline deficiency, one main trigger for nonalcoholic fatty liver disease (NAFLD), is closely related to lipid metabolism disorder. Previous study in a choline-deficient model has largely focused on gene expression rather than gene structure, especially sparse are studies regarding to alternative splicing (AS). In modern life science research, primary hepatocytes culture technology facilitates such studies, which can accurately imitate liver activity in vitro and show unique superiority. Whereas limitations to traditional hepatocytes culture technology exist in terms of efficiency and operability. This study pursued an optimization culture method for duck primary hepatocytes to explore AS in choline-deficient model. We performed an optimization culture method for duck primary hepatocytes with multi-step digestion procedure from Pekin duck embryos. Subsequently a NAFLD model was constructed with choline-free medium. RNA-seq and further analysis by rMATS were performed to identify AS events alterations in choline-deficency duck primary hepatocytes. The results showed E13 (embryonic day 13) to E15 is suitable to obtain hepatocytes, and the viability reached over 95% by trypan blue exclusion assay. Primary hepatocyte retained their biological function as well identified by Periodic Acid-Schiff staining method and Glucose-6-phosphate dehydrogenase activity assay, respectively. Meanwhile, genes of alb and afp and specific protein of albumin were detected to verify cultured hepatocytes. Immunofluorescence was used to evaluate purity of hepatocytes, presenting up to 90%. On this base, choline-deficient model was constructed and displayed significantly increase of intracellular triglyceride and cholesterol as reported previously. Intriguingly, our data suggested that AS events in choline-deficient model were implicated in pivotal biological processes as an aberrant transcriptional regulator, of which 16 genes were involved in lipid metabolism and highly enriched in glycerophospholipid metabolism. An effective and rapid protocol for obtaining duck primary hepatocytes was established, by which our findings manifested choline deficiency could induce the accumulation of lipid and result in aberrant AS events in hepatocytes, providing a novel insight into various AS in the metabolism role of choline.

Application of histochemical and immunohistochemical techniques for detection of lung tissue in cooked sausage.

Using unauthorized tissues in sausage is a common food adulteration in some parts of the world. This study was designed to compare the accuracy of histochemical and immunohistochemical techniques for the detection of lung tissue in cooked sausage samples. Samples with different levels of sheep lung tissues (1, 2.5, and 5%) and a control group were prepared and stained histochemically using H&E, Masson trichrome, and Periodic Acid-Schiff (PAS) stainings, and immunohistochemically using two different commercially-available antibodies of TTF1 Pan-cytokeratin. The highest positive results of lung tissue detection were achieved in sausage samples stained with anti-TTF1 immunohistochemical staining method. Both anti-TTF1 and anti-pan-cytokeratin immunohistochemical techniques detected all contaminated sausage samples treated with 50 g/kg lung tissues. Anti-TTF1 staining method had the highest odds ratio (7.4), followed by anti-pan-cytokeratin method (6.0). Reversely, PAS staining method had the lowest odds ratio (0.21), followed by Masson trichrome method (1.7). Additionally, anti-TTF1 method had the highest (1.8-31.0) confidence intervale (95%), while PAS had the lowest (0.02-2.1). Totally, the odds ratio of lung tissue detected by immunohistochemical methods were higher than those detected by histochemical staining. This is the first report on the comparison of histochemical and immunohistochemical techniques for lung tissue detection in cooked sausage. Anti-TTF1 immunohistochemical staining proved to be the most useful technique for the detection of unauthorized lung tissue in cooked sausages.

An Unexpected Guest in a Patient With Ulcerative Colitis

Question: A 16-year-old girl with a 2-year history of ulcerative colitis was referred to our university hospital because of fever, vomiting, and epigastric pain for 10 days; bloody diarrhea during the last month; and a rapid involuntary weight loss (5 kg). Sixth months earlier, during remission with oral mesalamine (60 mg/kg), she suffered a mild-to-moderate ulcerative colitis flare-up requiring oral prednisolone (1 mg/kg for 2 weeks and then tapered). At admission, physical examination showed pallor, mild signs of dehydration, and diffuse abdominal pain without signs of peritoneal irritation. Written consent for publication was obtained from the patient and her parents. Laboratory investigations revealed anemia, mild eosinophilia, hypoalbuminemia, an increased erythrocyte sedimentation rate, and fecal calprotectin. Specific serologic tests and blood and stool cultures for bacteria and viruses were negative. Colonoscopy showed pancolitis with continuous diffuse granular and friable mucosa (Mayo Score of 2) and upper endoscopy showed antral gastritis and diffuse duodenitis with nonbleeding erosions. The patient was started on steroids (1 mg/kg) followed by the amelioration of diarrhea, but persistence of mild grade fever and vomiting. Histologic examination showed crypt distortions and abscesses associated with goblet cell depletion. An upper endoscopy showed a normal esophageal and gastric mucosa. Duodenal histology (periodic acid–Schiff staining method) is presented in Figure A. What is the diagnosis? See the Gastroenterology web site (www.gastrojournal.org) for more information on submitting your favorite image to Clinical Challenges and Images in GI. Aspergillus fumigatus was identified in duodenal biopsy specimens. Serum galactomannan antigen test (Bio-rad Platelia Aspergillus Ag kit) and specific IgM (DRG IgM ELISA) assays were positive (index 4.1 and 6.5 UI/mL, respectively); chest and brain computed tomography scans were normal.1Donnelly J.P. Chen S.C. Kauffman C.A. et al.Revision and update of the consensus definitions of invasive fungal disease from the European Organization for Research and Treatment of Cancer and the Mycoses Study Group Education and Research Consortium.Clin Infect Dis. 2020; 71: 1367-1376Crossref PubMed Scopus (898) Google Scholar Antifungal treatment with voriconazole (loading dose: 6 mg/kg/12 h on the first day, then 4 mg/kg/12 h) was started and steroids were tapered.2Patterson T.F. Thompson 3rd, G.R. Denning D.W. et al.Practice guidelines for the diagnosis and management of aspergillosis: 2016 update by the Infectious Diseases Society of America.Clin Infect Dis. 2016; 63: e1-e60Crossref PubMed Scopus (1405) Google Scholar A comprehensive immune assessment was performed after discontinuing steroid therapy. After 1 week of treatment, the clinical condition and inflammatory markers improved. Aspergillus fumigatus is a ubiquitous saprophytic fungus with low pathogenicity in healthy individuals, but may be the major cause of life-threatening infection in immunocompromised individuals. Enteric aspergillosis is an uncommon presentation of invasive aspergillosis, which can disseminate hematogenously to other organs. Clinical features of invasive aspergillosis include abdominal pain, fever, and hematochezia or peritonitis subsequent to bowel ischemia or perforation; typical endoscopic findings include ulcerations or islands of necrotic tissue. Glucocorticosteroids are critical in the management of inflammatory bowel disease (IBD), but are associated with increased risk of opportunistic infections due to immune suppression. It should also be emphasized that IBD itself predisposes to infective complications3Kucharzik T. Ellul P. Greuter T. et al.ECCO guidelines on the prevention, diagnosis, and management of infections in inflammatory bowel disease.J Crohns Colitis. 2021; 15: 879-913Crossref PubMed Scopus (70) Google Scholar and that gastrointestinal infections can be potential triggers for IBD relapse.

HPMSCs inhibit the expression of PD-1 in CD4+IL-10+ T cells and mitigate liver damage in a GVHD mouse model by regulating the crosstalk between Nrf2 and NF-\u03baB signaling pathway

BackgroundThe activation of T cells and imbalanced redox metabolism enhances the development of graft-versus-host disease (GVHD). Human placenta-derived mesenchymal stromal cells (hPMSCs) can improve GVHD through regulating T cell responses. However, whether hPMSCs balance the redox metabolism of CD4+IL-10+ T cells and liver tissue and alleviate GVHD remains unclear. This study aimed to investigate the effect of hPMSC-mediated treatment of GVHD associated with CD4+IL-10+ T cell generation via control of redox metabolism and PD-1 expression and whether the Nrf2 and NF-κB signaling pathways were both involved in the process.MethodsA GVHD mouse model was induced using 6–8-week-old C57BL/6 and Balb/c mice, which were treated with hPMSCs. In order to observe whether hPMSCs affect the generation of CD4+IL-10+ T cells via control of redox metabolism and PD-1 expression, a CD4+IL-10+ T cell culture system was induced using human naive CD4+ T cells. The percentage of CD4+IL-10+ T cells and their PD-1 expression levels were determined in vivo and in vitro using flow cytometry, and Nrf2, HO-1, NQO1, GCLC, GCLM, and NF-κB levels were determined by western blotting, qRT-PCR, and immunofluorescence, respectively. Hematoxylin-eosin, Masson’s trichrome, and periodic acid-Schiff staining methods were employed to analyze the changes in hepatic tissue.ResultsA decreased activity of superoxide dismutase (SOD) and a proportion of CD4+IL-10+ T cells with increased PD-1 expression were observed in GVHD patients and the mouse model. Treatment with hPMSCs increased SOD activity and GCL and GSH levels in the GVHD mouse model. The percentage of CD4+IL-10+ T cells with decreased PD-1 expression, as well as Nrf2, HO-1, NQO1, GCLC, and GCLM levels, both in the GVHD mouse model and in the process of CD4+IL-10+ T cell generation, were also increased, but NF-κB phosphorylation and nuclear translocation were inhibited after treatment with hPMSCs, which was accompanied by improvement of hepatic histopathological changes.ConclusionsThe findings suggested that hPMSC-mediated redox metabolism balance and decreased PD-1 expression in CD4+IL-10+ T cells were achieved by controlling the crosstalk between Nrf2 and NF-κB, which further provided evidence for the application of hPMSC-mediated treatment of GVHD.

Indocyanine Green Uptake and Periodic Acid-Schiff Staining Method for Function Detection of Liver Cells are Affected by Different Cell Confluence.

The online version contains supplementary material available at 10.1007/s10616-021-00453-8.

Glandular structures in the major interlobular and extrapancreatic ducts in the guinea pig.

The morphology of the pancreatic duct system did not receive much attention as compared to the microanatomy of the exocrine and endocrine pancreas. The histological peculiarities of the excretory duct system are of major importance especially in laboratory animals like guinea pigs. The paper describes the histological peculiarities of the major interlobular and extrapancreatic ducts in guinea pigs. The pancreatic tissue samples were collected from five guinea pigs. For histological investigation, several pancreatic fragments underwent fixation in 10% buffered formalin and were later processed by the standard paraffin technique. Subsequentially, tissue sections were stained by Goldner's trichrome staining. The mucous substances were assessed by Alcian blue and Periodic acid-Schiff staining methods. The interlobular and main pancreatic ducts in the guinea pig present a simple columnar epithelium surrounded by a thick layer of dense connective tissue. The aforementioned epithelium of the main pancreatic ducts includes principal cells, goblet cells and basal cells. Additionally, the ductal epithelium presents occasional unicellular multiloculated intraepithelial mucous glands and prominent extraepithelial glands. The latter adopts a simple or compound tubular feature. The mucus elaborated by the three glandular types is mostly neutral in goblet cells, predominantly acidic in extraepithelial ductal glands, and a similar amount of acidic and neutral mucin in intraepithelial glands. In conclusion, the epithelium-associated mucous glands in the interlobular and main pancreatic ducts in the guinea pig are restricted not only to goblet cells. A substantial mucous discharge probably with a protecting role against irritative pancreatic juice derives from the main ductal intraepithelial and extraepithelial glands.

CHARACTERISTIC OF SKIN MORPHOLOGY OF SUNDA PORCUPINE (Hystrix javanica) WITH SPECIAL REFERENCE TO THE CONNECTIVE TISSUE

This study was conducted to investigate the histological characteristic, type, and distribution of connective tissue in Sunda porcupine skin. The investigation was carried out in three adult of sunda porcupines at microscopic level using hematoxylin eosin, Masson thrichrome, Verhoeffs van Gieson, alcian blue pH 2.5 and periodic acid Schiff staining methods. Skin consists of epidermis, dermis hypodermis, and subcutaneous muscle. Quill follicles were the main and dominant structure as well as the specific characteristic on Sunda porcupine skin. The connective tissue was distributed well in basal membrane, dermis, quill follicle, and hypodermis with various intensity and density. The collagen was the main fiber found in the skin while the elastin fiber was not observed. The acid carbohydrate was found distributed well in the skin while the neutral carbohydrate was not detected in this study. In addition the fibers of connective tissue associated with the adipose tissue which found plentifully in quill follicles and hypodermis. The present results showed that the wide distribution of connective tissue might have an important role on the wound healing physiology of Sunda porcupine skin.

Cutaneous myxidiosis in European eel, Anguilla anguilla (Linnaeus, 1758): histopathology, histochemistry and laminin immunohistochemistry.

Histopathology, histochemistry and immunohistochemistry of the integument of European eel, Anguilla anguilla (Linnaeus, 1758), infected by Myxidium sp. are reported. Skin samples from affected and unaffected eels were dissected, formalin fixed, paraffin embedded, sectioned and stained with H&E, Periodic acid-Schiff's staining method, Alcian Blue 8 GX pH 2.5/Periodic acid-Schiff's and McCallum-Goodpasture's Gram stain. Moreover, immunohistochemistry was performed using a primary polyclonal laminin antibody. Histologically, cysts (diameter 2-3mm) were observed mainly under the scale pockets, encircled by a thin collagen layer, lined by elongated, flattened fibroblasts and containing bipolar, PAS- and Gram-positive spores with opposite polar capsules. The epidermis stretched by the underlying cyst appeared dysplastic, thinned with a significant reduction in mucous cells number. Only inconsistent and aspecific inflammatory reaction was noted around the cysts at the dermis/epidermis interface. Intense laminin-like protein immunolabel was documented in the plasmodial ectoplasm and related to host anergia. This was the first report of laminin immunolabel in a member of the Myxozoa. Epidermal dysplasia represents likely an aspecific response against the underlying tensile force exerted by the developing parasite cyst, while fibroblast and collagen encapsulation denote a parasite-driven host response protecting, rather than harming, the encircled parasite.

Improved sensitivity for detection and quantitation of glycoproteins on polyacrylamide gels

A simple and sensitive technique for staining glycoproteins on polyacrylamide gels utilizing periodic acid-Schiff (PAS) reagent is described. The novelty of this technique is an additional incubation of the PAS stained gels in an ethanolic solution of metabisulfite. The treatment results in an about 20-fold increase in the intensitiy of bands and the color is stable for at least 1 week at room temperature. Furthermore, the integrated optical density of the bands is proportional to the amount of glycoproteins. Amounts as small as 0.2 μg of fetuin can be conveniently determined. As compared to other PAS staining methods, this procedure offers a remarkably improved sensitivity for detection and quantitation of glycoproteins following their electrophoretic separation on polyacrylamide gels.

Characterization of human exocrine pancreatic proteins by two-dimensional isoelectric focusing/sodium dodecyl sulfate gel electrophoresis

Exocrine proteins contained in human pancreatic juice were separated in two dimensions using isoelectric focusing and sodium dodecyl sulfate gel electrophoresis. Nineteen discrete proteins were found. Fifteen of these were identified by actual or potential enzyme activity and include three forms of trypsinogen, two forms each of procarboxypeptidase A, procarboxypeptidase B, proelastase, and colipase, and one form each for amylase, lipase, chymotrypsinogen, and prophospholipase A2. Lipase and four unidentified proteins were found to contain carbohydrate by the periodic acid Schiff staining method. Each pancreatic protein was characterized by isoelectric point and molecular weight. Proteins were quantitated according to relative mass, as measured by the incorporation of a mixture of 15 3H-amino acids into secretory proteins contained in tissue slices, and according to the distribution of Coomassie blue R stain among proteins contained in pancreatic juice, as determined by two-dimensional gel scanning and computer analysis. The second form of pancreatic procarboxypeptidase B (IEPn6.7) was present in only 4 of 10 subjects tested. Trypsinogens 1 and 3 were covalently labeled with 35SO4. Trypsin derived from trypsinogen 2 showed no inhibition with soybean trypsin inhibitor or Trasylol.

Correlative histologic study of the corpuscles of Stannius and the juxtaglomerular cells of teleost fishes

Different patterns of organization of corpuscles of Stannius and different types of distribution of juxtaglomerular cells are described in the kidneys of bony fishes. These patterns seem to bear little relationship to taxonomic position. Aldehyde-fuchsin and periodic acid-Schiff staining methods differentiate two kinds of granules in individual cells of the corpuscles of Stannius. Two kinds of cells may also exist in the corpuscles of Stannius of some species. The staining properties of granules in the corpuscles of Stannius and those in the juxtaglomerular cells are different. Juxtaglomerular cells were not found in Acipenser or in any of the Salmonidae examined but were present in all other teleosts. The location of juxtaglomerular cells on the afferent arterioles in many teleosts and their reaction to Bowie's stain suggest their homology with the juxtaglomerular cells of mammals. In fishes, it is conceivable that the juxtaglomerular cells and the corpuscles of Stannius together represent the renin-angiotensin-adrenal (zona glomerulosa) system.