Conversion to Sirolimus Ameliorates Cyclosporine-Induced Nephropathy in the Rat: Focus on Serum, Urine, Gene, and Protein Renal Expression Biomarkers

Protocols of conversion from cyclosporin A (CsA) to sirolimus (SRL) have been widely used in immunotherapy after transplantation to prevent CsA-induced nephropathy, but the molecular mechanisms underlying these protocols remain nuclear. This study aimed to identify the molecular pathways and putative biomarkers of CsA-to-SRL conversion in a rat model. Four animal groups (n=6) were tested during 9 weeks: control, CsA, SRL, and conversion (CsA for 3 weeks followed by SRL for 6 weeks). Classical and emergent serum, urinary, and kidney tissue (gene and protein expression) markers were assessed. Renal lesions were analyzed in hematoxylin and eosin, periodic acid-Schiff, and Masson’s trichrome stains. SRL-treated rats presented proteinuria and NGAL (serum and urinary) as the best markers of renal impairment. Short CsA treatment presented slight or even absent kidney lesions and TGF-β, NF-κβ, mTOR, PCNA, TP53, KIM-1, and CTGF as relevant gene and protein changes. Prolonged CsA exposure aggravated renal damage, without clear changes on the traditional markers, but with changes in serums TGF-βand IL-7, TBARs clearance, and kidney TGF-βand mTOR. Conversion to SRL prevented CsA-induced renal damage evolution (absent/mild grade lesions), while NGAL (serum versus urine) seems to be a feasible biomarker of CsA replacement to SRL.

Download Full-text

Tubular overexpression of Gremlin in transgenic mice aggravates renal damage in diabetic nephropathy

AJP Renal Physiology ◽

10.1152/ajprenal.00023.2015 ◽

2015 ◽

Vol 309 (6) ◽

pp. F559-F568 ◽

Cited By ~ 24

Author(s):

Vanessa Marchant ◽

Alejandra Droguett ◽

Graciela Valderrama ◽

M. Eugenia Burgos ◽

Daniel Carpio ◽

...

Keyword(s):

Diabetic Nephropathy ◽

Renal Damage ◽

Transforming Growth Factor ◽

Mesangial Cells ◽

Interstitial Fibrosis ◽

Smooth Muscle Actin ◽

Periodic Acid ◽

Kidney Tissue ◽

Renal Tubules ◽

Periodic Acid Schiff

Diabetic nephropathy (DN) is currently a leading cause of end-stage renal failure worldwide. Gremlin was identified as a gene differentially expressed in mesangial cells exposed to high glucose and in experimental diabetic kidneys. We have described that Gremlin is highly expressed in biopsies from patients with diabetic nephropathy, predominantly in areas of tubulointerstitial fibrosis. In streptozotocin (STZ)-induced experimental diabetes, Gremlin deletion using Grem1 heterozygous knockout mice or by gene silencing, ameliorates renal damage. To study the in vivo role of Gremlin in renal damage, we developed a diabetic model induced by STZ in transgenic (TG) mice expressing human Gremlin in proximal tubular epithelial cells. The albuminuria/creatinuria ratio, determined at week 20 after treatment, was significantly increased in diabetic mice but with no significant differences between transgenic (TG/STZ) and wild-type mice (WT/STZ). To assess the level of renal damage, kidney tissue was analyzed by light microscopy (periodic acid-Schiff and Masson staining), electron microscopy, and quantitative PCR. TG/STZ mice had significantly greater thickening of the glomerular basement membrane, increased mesangial matrix, and podocytopenia vs. WT/STZ. At the tubulointerstitial level, TG/STZ showed increased cell infiltration and mild interstitial fibrosis. In addition, we observed a decreased expression of podocin and overexpression of monocyte chemoattractant protein-1 and fibrotic-related markers, including transforming growth factor-β1, Col1a1, and α-smooth muscle actin. Together, these results show that TG mice overexpressing Gremlin in renal tubules develop greater glomerular and tubulointerstitial injury in response to diabetic-mediated damage and support the involvement of Gremlin in diabetic nephropathy.

Download Full-text

The potential renal toxicity of silver nanoparticles after repeated oral exposure and its underlying mechanisms

BMC Nephrology ◽

10.1186/s12882-021-02428-5 ◽

2021 ◽

Vol 22 (1) ◽

Author(s):

Hamed Nosrati ◽

Manijeh Hamzepoor ◽

Maryam Sohrabi ◽

Massoud Saidijam ◽

Mohammad Javad Assari ◽

...

Keyword(s):

Silver Nanoparticles ◽

Molecular Mechanisms ◽

Renal Toxicity ◽

Periodic Acid ◽

Critical Role ◽

Oral Exposure ◽

Control Group ◽

Rt Pcr ◽

Periodic Acid Schiff

Abstract Background Silver nanoparticles (AgNPs) can accumulate in various organs after oral exposure. The main objective of the current study is to evaluate the renal toxicity induced by AgNPs after repeated oral exposure and to determine the relevant molecular mechanisms. Methods In this study, 40 male Wistar rats were treated with solutions containing 30, 125, 300, and 700 mg/kg of AgNPs. After 28 days of exposure, histopathological changes were assessed using hematoxylin-eosin (H&E), Masson’s trichrome, and periodic acid-Schiff (PAS) staining. Apoptosis was quantified by TUNEL and immunohistochemistry of caspase-3, and the level of expression of the mRNAs of growth factors was determined using RT-PCR. Results Histopathologic examination revealed degenerative changes in the glomeruli, loss of tubular architecture, loss of brush border, and interrupted tubular basal laminae. These changes were more noticeable in groups treated with 30 and 125 mg/kg. The collagen intensity increased in the group treated with 30 mg/kg in both the cortex and the medulla. Apoptosis was much more evident in middle-dose groups (i.e., 125 and 300 mg/kg). The results of RT-PCR indicated that Bcl-2 and Bax mRNAs upregulated in the treated groups (p < 0.05). Moreover, the data related to EGF, TNF-α, and TGF-β1 revealed that AgNPs induced significant changes in gene expression in the groups treated with 30 and 700 mg/kg compared to the control group. Conclusion Our observations showed that AgNPs played a critical role in in vivo renal toxicity.

Download Full-text

Deep learning-based transformation of H&E stained tissues into special stains

Nature Communications ◽

10.1038/s41467-021-25221-2 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Kevin de Haan ◽

Yijie Zhang ◽

Jonathan E. Zuckerman ◽

Tairan Liu ◽

Anthony E. Sisk ◽

...

Keyword(s):

Visual Inspection ◽

Kidney Diseases ◽

Periodic Acid ◽

Needle Core Biopsy ◽

Silver Stain ◽

Periodic Acid Schiff ◽

Tissue Sections ◽

Biopsy Tissue ◽

Hematoxylin And Eosin

AbstractPathology is practiced by visual inspection of histochemically stained tissue slides. While the hematoxylin and eosin (H&E) stain is most commonly used, special stains can provide additional contrast to different tissue components. Here, we demonstrate the utility of supervised learning-based computational stain transformation from H&E to special stains (Masson’s Trichrome, periodic acid-Schiff and Jones silver stain) using kidney needle core biopsy tissue sections. Based on the evaluation by three renal pathologists, followed by adjudication by a fourth pathologist, we show that the generation of virtual special stains from existing H&E images improves the diagnosis of several non-neoplastic kidney diseases, sampled from 58 unique subjects (P = 0.0095). A second study found that the quality of the computationally generated special stains was statistically equivalent to those which were histochemically stained. This stain-to-stain transformation framework can improve preliminary diagnoses when additional special stains are needed, also providing significant savings in time and cost.

Download Full-text

Autofluorescence microscopy as a label-free tool for renal histology and glomerular segmentation

10.1101/2021.07.16.452703 ◽

2021 ◽

Author(s):

Nathan Heath Patterson ◽

Elizabeth K Neumann ◽

Kavya Sharman ◽

Jamie L Allen ◽

Raymond C Harris ◽

...

Keyword(s):

Recognition Performance ◽

Periodic Acid ◽

Kidney Tissue ◽

Human Kidney ◽

True Positive Rate ◽

Label Free ◽

Periodic Acid Schiff ◽

Renal Histology ◽

Fresh Frozen ◽

Histological Staining

Functional tissue units (FTUs) composed of multiple cells like the glomerulus in the kidney nephron play important roles in health and disease. Histological staining is often used for annotation or segmentation of FTUs, but chemical stains can introduce artefacts through experimental factors that influence analysis. Secondly, many molecular -omics techniques are incompatible with common histological stains. To enable FTU segmentation and annotation in human kidney without the need for histological staining, we detail here the use of widefield autofluorescence (AF) microscopy as a simple, label-free modality that provides detailed renal morphology comparable to periodic acid-Schiff (PAS) stained tissue in both formalin-fixed paraffin-embedded (FFPE) and fresh frozen samples and with no tissue processing beyond sectioning. We demonstrate automated deep learning-based glomerular unit recognition and segmentation on PAS and AF images of the same tissue section from 9 fresh frozen samples and 9 FFPE samples. All training comparisons were carried out using registered AF microscopy and PAS stained whole slide images originating from the same section, and the recognition models were built with the exact same training and test examples. Measures of recognition performance, such as the Dice-Sorensen coefficient, the true positive rate, and the positive predictive value differed less than 2% between standard PAS and AF microscopy for both preservation methods. These results demonstrate that AF is a potentially powerful tool to study human kidney tissue, that it can serve as a label-free source for automated and manual annotation of tissue structures.

Download Full-text

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

Malaysian Journal of Medical Sciences ◽

10.21315/mjms2020.27.5.6 ◽

2020 ◽

Vol 27 (5) ◽

pp. 53-61

Author(s):

Abdullah Saleh Alkhamiss

Keyword(s):

Helicobacter Pylori ◽

Alcian Blue ◽

National Guard ◽

Periodic Acid ◽

Giemsa Stain ◽

Periodic Acid Schiff ◽

Gastric Biopsies ◽

Hematoxylin And Eosin ◽

H Pylori ◽

Sensitivity Specificity

Background: This study was undertaken to evaluate the preferred method (Giemsa or periodic acid Schiff-Alcian blue [PAS-AB] stains) of detecting Helicobacter pylori (H. pylori) 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 H. pylori 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 H. pylori, 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 H. pylori infection in general and in 13 of them, active gastritis cases were discovered. Fourteen out of these 15 H. pylori 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 H. pylori detection. Conclusion: Giemsa stain has better sensitivity and specificity in gastric H. pylori infection detection than PAS-AB. Therefore, using PAS-AB stain to detect H. pylori infection is not recommended.

Download Full-text

Effects of parathyroid extract. II: changes preceding renal calcification

American Journal of Physiology-Legacy Content ◽

10.1152/ajplegacy.1962.203.4.676 ◽

1962 ◽

Vol 203 (4) ◽

pp. 676-680 ◽

Cited By ~ 4

Author(s):

Reagan H. Bradford ◽

R. Palmer Howard ◽

Walter Joel ◽

Jerry Puls ◽

M. R. Shetlar

Keyword(s):

Serum Albumin ◽

Periodic Acid ◽

Blue Staining ◽

Alcian Blue Staining ◽

Periodic Acid Schiff ◽

Protein Ratio ◽

Parathyroid Extract ◽

Staining Techniques ◽

Consistent Increase ◽

Hematoxylin And Eosin

Parathyroid extract, a total of 860 units, has been administered to rats in small, progressively increasing doses over a period of 12 days. The effects on serum protein, total glycoprotein, glycoprotein/protein ratio, calcium, individual protein and glycoprotein fractions, and renal calcification have been presented. Kidney sections from each rat were studied by histochemical techniques for calcification, neutral polysaccharide, and acid mucopolysaccharide. The serum total glycoprotein, glycoprotein/protein ratio, and calcium were found to be elevated. The serum albumin was decreased, whereas the globulin fractions were essentially unchanged. The globulin glycoprotein hexose, mostly α1-globulin, was increased; albumin glycoprotein hexose showed a somewhat less consistent increase. A "precalcification" periodic acid-Schiff-staining intraluminal material was demonstrated in the kidney after parathyroid extract treatment for 4 days. This was followed approximately 2 days later by Alcian blue-staining material and calcification demonstrable by hematoxylin and eosin and by Kóssa staining techniques. This has been interpreted as suggesting a glycoprotein-containing lesion which precedes calcification.

Download Full-text

Deep Learning-Based Spermatogenic Staging Assessment for Hematoxylin and Eosin-Stained Sections of Rat Testes

Toxicologic Pathology ◽

10.1177/0192623320969678 ◽

2020 ◽

pp. 019262332096967

Author(s):

Dianne M. Creasy ◽

Satish T. Panchal ◽

Rohit Garg ◽

Pranab Samanta

Keyword(s):

Deep Learning ◽

Digital Images ◽

Periodic Acid ◽

Wistar Rat ◽

Specialized Training ◽

Periodic Acid Schiff ◽

Automated Method ◽

Histopathological Evaluation ◽

Hematoxylin And Eosin ◽

Sensitive Method

In preclinical toxicology studies, a “stage-aware” histopathological evaluation of testes is recognized as the most sensitive method to detect effects on spermatogenesis. A stage-aware evaluation requires the pathologist to be able to identify the different stages of the spermatogenic cycle. Classically, this evaluation has been performed using periodic acid-Schiff (PAS)-stained sections to visualize the morphology of the developing spermatid acrosome, but due to the complexity of the rat spermatogenic cycle and the subtlety of the criteria used to distinguish between the 14 stages of the cycle, staging of tubules is not only time consuming but also requires specialized training and practice to become competent. Using different criteria, based largely on the shape and movement of the elongating spermatids within the tubule and pooling some of the stages, it is possible to stage tubules using routine hematoxylin and eosin (H&E)-stained sections, thereby negating the need for a special PAS stain. These criteria have been used to develop an automated method to identify the stages of the rat spermatogenic cycle in digital images of H&E-stained Wistar rat testes. The algorithm identifies the spermatogenic stage of each tubule, thereby allowing the pathologist to quickly evaluate the testis in a stage-aware manner and rapidly calculate the stage frequencies.

Download Full-text

Pathological Findings in the Pituitary Glands of Dogs and Cats

Veterinary Pathology ◽

10.1177/0300985818784162 ◽

2018 ◽

Vol 55 (6) ◽

pp. 880-888 ◽

Cited By ~ 8

Author(s):

Laura Polledo ◽

Guy C. M. Grinwis ◽

Peter Graham ◽

Mark Dunning ◽

Kerstin Baiker

Keyword(s):

Adrenocorticotropic Hormone ◽

Periodic Acid ◽

Clinical Manifestations ◽

Histopathological Diagnosis ◽

Periodic Acid Schiff ◽

Melanocyte Stimulating Hormone ◽

Nodular Hyperplasia ◽

Cystic Changes ◽

Pituitary Glands ◽

Hematoxylin And Eosin

With the exception of classic functional adenomas in dogs and horses, pituitary lesions are infrequently described in the veterinary literature. Approximately 10% of pituitary glands from asymptomatic humans contain abnormalities, but the equivalent proportion in small animals is unknown. Pituitary glands from 136 dogs and 65 cats collected during routine necropsies were examined to determine the prevalence of pituitary lesions and their histopathological diagnosis. Lesions were characterized in sections stained with hematoxylin and eosin, periodic acid-Schiff (PAS), Gordon and Sweet’s and reticulin stains, and immunohistochemistry for adrenocorticotropic hormone (ACTH), growth hormone, melanocyte stimulating hormone–α, and prolactin. Pituitary abnormalities were identified in 36 of 136 (26.4%) dogs and 10 of 65 (15.3%) cats. Cystic changes were the most common lesion, occurring in 18 (13.2%) dogs and 8 (12.3%) cats. Pituitary neoplasia was detected in 14.1% (12/85) of middle-aged and old dogs; 1 (1.5%) cat had pituitary nodular hyperplasia. PAS and reticulin stains helped differentiate ACTH-immunoreactive adenomas from hyperplastic nodules: adenomas contained PAS-positive intracytoplasmic granules and loss of the normal reticulin network. One dog had a pituitary carcinoma with infiltration into the thalamus. Other pituitary abnormalities included secondary metastases (2 dogs) and hypophysitis (4 dogs, 1 cat). In most cases, the lesion appeared to be subclinical and could be considered incidental, whereas clinical manifestations were apparent in only 4 dogs (2.9%) and none of the cats with pituitary lesions. Pituitary abnormalities are common in dogs and cats, and their clinical relevance requires further investigation.

Download Full-text

Condrolipoma na orelha de um cão

Acta Scientiae Veterinariae ◽

10.22456/1679-9216.85347 ◽

2017 ◽

Vol 45 ◽

pp. 4

Author(s):

Maria De Fátima De Souza ◽

Alinne Emanuelle Monteiro Sarmento ◽

Francisca Maria Sousa Barbosa ◽

Raul Antunes Silva Siqueira ◽

Ivia Carmem Talieri ◽

...

Keyword(s):

Alcian Blue ◽

Periodic Acid ◽

Fibrous Tissue ◽

Rare Event ◽

Periodic Acid Schiff ◽

Non Invasive ◽

Pas Staining ◽

Staining Techniques ◽

Hematoxylin And Eosin ◽

Well Differentiated

Background: Lipoma is defined as a tumor composed of mature adipocytes, identical to the cells that form normal adult fat. Other elements may also be present, such as cartilaginous tissue. Chondrolipoma is characterized by the presence of differentiated cartilage islands spread within a typical lipoma, in which mucin and collagen can be observed around isles of chondrocytes. This condition affects mainly middle-aged or elderly dogs as well as elderly humans. In the scientific literature, chondrolipomas have not yet been described on the ear of dogs. Therefore, this study aimed to report a case of chondrolipoma on the ear of a dog and its histopathological and histochemical description.Case: A female, 10-year-old, Labrador retriever dog was admitted to the Veterinarian Hospital of the Federal University of Paraíba (UFPB), Areia, Brazil. The dog was presented with an ulcerated pendulous nodule on the ear, with a clinical evolution of approximately 7 months. A biopsy sample was taken and submitted to the Veterinary Histopathology Laboratory of the UFPB. Macroscopic evaluation revealed a pendulous and firm nodule, with a round apex, measuring 1.3cm in diameter. The epidermis covering the nodule exhibited an ulcer of 0.5cm in length. The nodule exhibited resistance to cutting and the surface was dark red with hard areas in the center. Sample was fixed in 10% formalin and routinely processed for histology, embedded in paraffin, and stained with hematoxylin and eosin (HE), alcian blue (AB), Masson’s trichrome (MT), and periodic acid-Schiff (PAS). Histopathological evaluation revealed that the nodule was encapsulated, formed by adipocytes of variable sizes. Adipocytes were well-differentiated and the nodule was defined as lipoma. Within the lipoma there was proliferation of chondrocyte islands in a bluish basophilic matrix. Adjacently, abundant fibrous connective tissue was found intertwined with blood vessels. Some vessels exhibited red blood cells in the vicinity, indicating hemorrhage. The epidermis covering the nodule was slightly acanthotic and moderately hyperkeratotic. MT staining marked the fibrous tissue in blue. AB staining revealed blue areas confirming the presence of mucin around cartilage islands. PAS staining revealed the presence of glycogen between mesenchymal cells, which included chondrocytes.Discussion: The diagnosis of chondrolipoma was concluded with the presence of a well-differentiated lipoma, randomly intertwined with cartilage islands, which were also well-differentiated and spread within a typical lipoma. Additionally, the presence of mucin and collagen around the isles of chondrocytes facilitated the diagnosis. Chondrocytes presented different phases of maturation. The case described here exhibited microscopic characteristics that were similar but somewhat different to those previously reported by other authors, in considering that the nodule was encapsulated, and did not exhibit cell atypia or mineralization in the cartilage islands. Most studies published used PAS and Masson’s trichrome staining techniques to confirm the diagnosis of chondrolipoma. However, alcian blue also aids in diagnosing this condition. The tumor developed for months before the dog was taken to a veterinarian, when it finally became ulcerated, which characterized it as non-invasive. Chondrolipoma is a benign tumor, which does not induce metastasis and once excised does not exhibit recurrence. In veterinary medicine, the presence of cartilage within a lipoma is a rare event.

Download Full-text

Submucosal Lifting Agent ORISE Gel Remnants Histopathologically Mimic Mucin and Malignancy: A Case Series

American Journal of Clinical Pathology ◽

10.1093/ajcp/aqz113.088 ◽

2019 ◽

Vol 152 (Supplement_1) ◽

pp. S73-S73 ◽

Cited By ~ 2

Author(s):

Luke Cypher ◽

Shaoli Sun ◽

Erin Forster ◽

Brenda Hoffman ◽

David Lewin

Keyword(s):

Colon Cancer ◽

Epithelial Cells ◽

Amorphous Material ◽

Periodic Acid ◽

Case Series ◽

The United States ◽

Periodic Acid Schiff ◽

Good Communication ◽

Gastrointestinal Lesions ◽

Hematoxylin And Eosin

Abstract Introduction There were an estimated 18.1 million new cancer cases in 2018, with colon cancer being the third most common worldwide. Colon cancer development is an accumulation of mutations resulting in normal epithelial cells transforming into adenomas and then adenocarcinomas. In certain scenarios, endoscopic interventions have gained considerable momentum over invasive surgery as an alternative to manage early gastrointestinal lesions. New techniques such as endoscopic mucosal resection (EMR) and endoscopic submucosal dissection allow for removal of large, flat sessile polyps. Successful EMR is dependent on expanding the submucosal space to create adequate lift of the polyp to facilitate tissue capture and to avoid perforation and excess bleeding. ORISE gel (Boston Scientific) is a submucosal lifting agent currently in use in the United States. Methods We present three cases of gastrointestinal specimens obtained using ORISE gel. Histological analysis with hematoxylin and eosin revealed submucosal amorphous deposits that appeared to be mucin. Due to the concern for malignancy, additional stains were performed, including periodic acid–Schiff with diastase digestion (DPAS) to identify mucin. DPAS staining for mucin was negative, indicating the mucinous-appearing amorphous material seen on hematoxylin and eosin staining was not mucin but a likely remnant ORISE gel used during EMR. Additional immunohistochemical stains for epithelial cells (cytokeratin AE1/AE3) were also performed to exclude the presence of infiltrating tumor cells. Conclusion These three cautionary cases reveal the importance of good communication between endoscopists and pathology. In an effort to avoid overdiagnosis and/or the usage of unnecessary additional stains, pathologists should be alerted of ORISE gel usage.

Download Full-text