PAS Staining of Fresh Frozen or Paraffin Embedded Human Kidney Tissue v2 (protocols.io.buaknscw)

protocols.io ◽  
2021 ◽  
Author(s):  
Elizabeth Neumann ◽  
Jamie Allen ◽  
Jennifer Harvey ◽  
Maya Brewer ◽  
Carrie not provided ◽  
...  
Keyword(s):  
Kidney Tissue ◽  
Human Kidney ◽  
Fresh Frozen ◽  
Pas Staining

scholarly journals PAS Staining of Fresh Frozen or Paraffin Embedded Human Kidney Tissue v1 (protocols.io.4qngvve)

protocols.io ◽  
2019 ◽  
Author(s):  
Jamie Allen ◽  
Jennifer Harvey ◽  
Maya Brewer ◽  
Mark de ◽  
Jeff Spraggins
Keyword(s):  
Kidney Tissue ◽  
Human Kidney ◽  
Fresh Frozen ◽  
Pas Staining

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

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.

CORTICOSTEROIDS IN HUMAN KIDNEY TISSUE

1969 ◽  
Vol 61 (1_Suppl) ◽  
pp. S66
Author(s):  
A. T. A. Fazekas ◽  
I. Gy. Fazekas
Keyword(s):  
Kidney Tissue ◽  
Human Kidney

scholarly journals The Globoseries Glycosphingolipid Sialosyl Galactosyl Globoside Is Found in Urinary Tract Tissues and Is a Preferred Binding Receptor In Vitro for Uropathogenic Escherichia coli Expressing pap-Encoded Adhesins

1998 ◽  
Vol 66 (8) ◽  
pp. 3856-3861 ◽  
Author(s):  
A. E. Stapleton ◽  
M. R. Stroud ◽  
S. I. Hakomori ◽  
W. E. Stamm
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract ◽  
Kidney Tissue ◽  
Human Kidney ◽  
Blood Group Antigens ◽  
E Coli ◽  
Vaginal Epithelial Cells ◽  
History Of ◽  
Tract Infections

ABSTRACT Women with a history of recurrent Escherichia coliurinary tract infections (UTIs) are significantly more likely to be nonsecretors of blood group antigens than are women without such a history, and vaginal epithelial cells (VEC) from women who are nonsecretors show enhanced adherence of uropathogenic E. coli isolates compared with cells from secretors. We previously extracted glycosphingolipids (GSLs) from native VEC and determined that nonsecretors (but not secretors) selectively express two extended globoseries GSLs, sialosyl galactosyl globoside (SGG) and disialosyl galactosyl globoside (DSGG), which specifically bound uropathogenicE. coli R45 expressing a P adhesin. In this study, we demonstrated, by purifying the compounds from this source, that SGG and DSGG are expressed in human kidney tissue. We also demonstrated that SGG and DSGG isolated from human kidneys bind uropathogenic E. coli isolates expressing each of the three classes ofpap-encoded adhesins, including cloned isolates expressing PapG from J96, PrsG from J96, and PapG from IA2, and the wild-type isolates IA2 and R45. We metabolically 35S labeled these five E. coli isolates and measured their relative binding affinities to serial dilutions of SGG and DSGG as well as to globotriaosylceramide (Gb3) and globotetraosylceramide (Gb4), two other globoseries GSLs present in urogenital tissues. Each of the five E. coli isolates bound to SGG with the highest apparent avidity compared with their binding to DSGG, Gb3, and Gb4, and each isolate had a unique pattern of GSL binding affinity. These studies further suggest that SGG likely plays an important role in the pathogenesis of UTI and that its presence may account for the increased binding of E. colito uroepithelial cells from nonsecretors and for the increased susceptibility of nonsecretors to recurrent UTI.

scholarly journals O32 Ontogeny of human kidney OCT2 expression across the paediatric age range

2019 ◽  
Vol 104 (6) ◽  
pp. e14.2-e14
Author(s):  
N Smeets ◽  
B van Groen ◽  
J Pertijs ◽  
M Wilmer ◽  
B Smeets ◽  
...  
Keyword(s):  
Organic Cation ◽  
Basolateral Membrane ◽  
Age Groups ◽  
Kidney Tissue ◽  
Human Kidney ◽  
Organic Cation Transporter ◽  
Age Related ◽  
Paediatric Age ◽  
Organic Cation Transporter 2 ◽  
Related Pattern

BackgroundIn adults, the organic cation transporter 2 (protein name OCT2, gene name SLC22A2) is localised in the kidney proximal tubules where it mediates organic cation secretion. Hence, the transporter plays a role in the disposition and excretion of several drugs and drug-drug interactions. To better understand the disposition of OCT2 substrate drugs in children, we studied OCT2 localisation and expression in paediatric kidney tissue.MethodsThe expression of OCT2 was visualised in tissue using immunohistochemical staining. Tissues were derived post-mortem from children aged 0 -14 years. Gestational age varied between 24 and 40 weeks. Intensity of the staining at the basolateral membrane was scored by two individual observers using three categories; negative, detectible and high. Agreement between two observers was determined using Cohen’s kappa.Results44 kidney samples (n=17 neonates, n=17 infants, n=7 children, n=3 adolescent) were analysed and scored. There was substantial agreement between two judgements with a kappa of 0.773 (p< 0.005). No age related pattern was observed in the expression of OCT2. Even in the youngest age group, the expression of OCT2 was clearly visible.ConclusionThe kidney expression of OCT2 did not show an age-related pattern. In all age groups, expression levels were similar and OCT2 was properly localised at the basolateral membrane. These findings suggest that, with increasing age, OCT2 will not influence the renal excretion of its substrates.Disclosure(s)Nothing to disclose

Antigenic structure of human kidney tissue

1969 ◽  
Vol 67 (6) ◽  
pp. 665-668
Author(s):  
R. F. Averkina ◽  
O. E. Vyazov
Keyword(s):  
Kidney Tissue ◽  
Human Kidney ◽  
Antigenic Structure

scholarly journals A role for OCRL in glomerular function and disease

2019 ◽  
Vol 35 (4) ◽  
pp. 641-648 ◽  
Author(s):  
Rebecca Preston ◽  
Richard W Naylor ◽  
Graham Stewart ◽  
Agnieszka Bierzynska ◽  
Moin A Saleem ◽  
...  
Keyword(s):  
In Situ Hybridisation ◽  
Kidney Tissue ◽  
Human Kidney ◽  
Tubular Function ◽  
Glomerular Function ◽  
Persistent Proteinuria ◽  
Whole Exome ◽  
Proximal Tubulopathy ◽  
Renal Phenotype

Abstract Background Lowe syndrome and Dent-2 disease are caused by mutations in the OCRL gene, which encodes for an inositol 5-phosphatase. The renal phenotype associated with OCRL mutations typically comprises a selective proximal tubulopathy, which can manifest as Fanconi syndrome in the most extreme cases. Methods Here, we report a 12-year-old male with nephrotic-range proteinuria and focal segmental glomerulosclerosis on renal biopsy. As a glomerular pathology was suspected, extensive investigation of tubular function was not performed. Results Surprisingly, whole exome sequencing identified a genetic variant in OCRL (c1467-2A>G) that introduced a novel splice mutation leading to skipping of exon 15. In situ hybridisation of adult human kidney tissue and zebrafish larvae showed OCRL expression in the glomerulus, supporting a role for OCRL in glomerular function. In cultured podocytes, we found that OCRL associated with the linker protein IPIP27A and CD2AP, a protein that is important for maintenance of the podocyte slit diaphragm. Conclusion Taken together, this work suggests a previously under-appreciated role for OCRL in glomerular function and highlights the importance of investigating tubular function in patients with persistent proteinuria.

ENGRAFTMENT OF HUMAN KIDNEY TISSUE IN RAT RADIATION CHIMERA

1997 ◽  
Vol 64 (11) ◽  
pp. 1541-1550 ◽  
Author(s):  
Benjamin Dekel ◽  
Tatyana Burakova ◽  
Hadar Marcus ◽  
Elias Shezen ◽  
Sylvie Polack ◽  
...  
Keyword(s):  
Kidney Tissue ◽  
Human Kidney ◽  
Radiation Chimera

scholarly journals Characterization of the Hoechst 33342 side population from normal and malignant human renal epithelial cells

2008 ◽  
Vol 295 (3) ◽  
pp. F680-F687 ◽  
Author(s):  
Sanjai K. Addla ◽  
Mick D. Brown ◽  
Claire A. Hart ◽  
Vijay A. C. Ramani ◽  
Noel W. Clarke
Keyword(s):  
Stem Cell ◽  
Epithelial Cells ◽  
Side Population ◽  
Kidney Tissue ◽  
Human Kidney ◽  
Proliferative Potential ◽  
Stem Cell Markers ◽  
Hoechst 33342 ◽  
Tissue Samples

The fundamental changes which predispose for renal cell carcinoma (RCC) are poorly characterized. It is hypothesized that “cancer stem cells” may be influential in carcinogenesis, and the epithelial side population (SP) is enriched for stemlike cells in other epithelial cancers. In this study, we have isolated and characterized the SP and non-SP (NSP) populations from normal (NK) and malignant (RCC) human kidney tissue. NK specimens were taken from patients undergoing non-renal cancer surgery and paired malignant and macroscopically normal tissue samples were taken from patients undergoing surgery for RCC. The Hoechst 33342 dye efflux technique was used to isolate epithelial SP and NSP from normal and malignant human renal tissue. Cellular subpopulations were phenotyped for lineage, cell cycle, and putative stem cell markers, and functionally characterized using in vitro colony-forming and proliferation assays. The SP constituted 3.8 ± 0.4 and 5.9 ± 0.9% of epithelial cells in NK and RCC, respectively, of which 14.1 ± 3.5 and 13.2 ± 3.6% were shown to be in G0. SP cells demonstrated greater proliferative potential in colony-forming efficiency, long-term culture, and spheroids assays and were shown to be maintained upon tissue culture passage. We have shown that the renal SP is enriched for quiescent cells, with a high proliferative capacity and stemlike properties. The population is, however, heterogeneous, confirming that the terms “SP cell” and “stem cell” cannot be used interchangeably.

scholarly journals Human endogenous retrovirus transcription profiles of the kidney and kidney-derived cell lines

2011 ◽  
Vol 92 (10) ◽  
pp. 2356-2366 ◽  
Author(s):  
Sonja Haupt ◽  
Michele Tisdale ◽  
Michelle Vincendeau ◽  
Mary Anne Clements ◽  
David T. Gauthier ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Line ◽  
Cell Lines ◽  
Kidney Tissue ◽  
Human Kidney ◽  
Endogenous Retrovirus ◽  
Endogenous Retroviruses ◽  
Human Endogenous Retrovirus ◽  
Tissue Samples ◽  
Transcription Pattern

The human genome comprises approximately 8–9 % of human endogenous retroviruses (HERVs) that are transcribed with tissue specificity. However, relatively few organs have been examined in detail for individual differences in HERV transcription pattern, nor have tissue-to-cell culture comparisons been frequently performed. Using an HERV-specific DNA microarray, a core HERV transcription profile was established for the human kidney comparing 10 tissue samples. This core represents HERV groups expressed uniformly or nearly so in non-tumour kidney tissue. The profiles obtained from non-tumour tissues were compared to 10 renal tumour tissues (renal cell carcinoma, RCC) derived from the same individuals and additionally, to 22 RCC cell lines. No RCC cell line or tumour-specific differences were observed, suggesting that HERV transcription is not altered in RCC. However, when comparing tissue transcription to cell line transcription, there were consistent differences. The differences were irrespective of cancer state and included cell lines derived from non-tumour kidney tissue, suggesting that a specific alteration of HERV transcription occurs when establishing cell lines. In contrast to previous publications, all known HERV-derived tumour antigens, including those identified in RCC, were expressed both in multiple RCC cell lines and several non-tumour tissue-derived cell lines, a result that contrasts with findings from patient samples. The results establish the core kidney transcription pattern of HERVs and reveal differences between cell culture lines and tissue samples.

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