scholarly journals Meso Scale Discovery and Luminex Comparative Analysis of Calbindin D28K

2009 ◽  
Vol 2009 ◽  
pp. 1-5 ◽  
Author(s):  
Samer Sourial ◽  
Maritha Marcusson-Ståhl ◽  
Karin Cederbrant
Keyword(s):  
Comparative Analysis ◽  
Human Kidney ◽  
Distal Tubule ◽  
Specific Protein ◽  
Tubular Damage ◽  
Meso Scale Discovery ◽  
Multiplex Immunoassay ◽  
Pathological Conditions ◽  
Calbindin D28k ◽  
Meso Scale

The sensitivity of different renal regions to xenobiotics requires the development of a multiplex immunoassay for the simultaneous analysis of kidney biomarkers. Calbindin D28K is a distal tubule-specific protein that can be detected in urine under pathological conditions. In this study, a pair of anti-calbindin D28K antibodies was used in an immunoassay for the detection of calbindin D28K expression in rat and human kidney and urine. Comparative analysis of the immunoassay was performed on the Meso Scale Development (MSD) and Luminex platforms. Analysis on both platforms detected calbindin D28K concentrations between 100 ng/mL and 100 pg/mL. Luminex detected 10-fold the amount of calbindin D28K in samples analyzed as compared to MSD, whereas calbindin D28K level in rat and human urine was below detection limit in both platforms. The application of the immunoassays described herein may be useful in toxicological and pathological studies of distal tubular damage in rats and human.

Immunolocalization of calcitriol receptor, 24-hydroxylase cytochrome P-450, and calbindin D28k in human kidney

1994 ◽  
Vol 266 (3) ◽  
pp. F477-F485 ◽  
Author(s):  
R. Kumar ◽  
J. Schaefer ◽  
J. P. Grande ◽  
P. C. Roche
Keyword(s):  
Vitamin D ◽  
Proximal Tubule ◽  
Polyclonal Antibodies ◽  
Collecting Duct ◽  
Human Kidney ◽  
Distal Tubule ◽  
Calbindin D28k ◽  
Parietal Epithelial Cells ◽  
Cytochrome P 450 ◽  
In Cells

The precise localization of the calcitriol (1 alpha,25-dihydroxyvitamin D3) receptor (VDR) and the 25-hydroxyvitamin D3 [25(OH)D3] 24-hydroxylase cytochrome P-450 in the human kidney is unknown. Using newly developed polyclonal antibodies against the human VDR, we demonstrate that the receptor is present in cells of the distal tubule, the collecting duct, the proximal tubule, and in the parietal epithelial cells of the glomerulus. In the distal tubule and collecting duct not all cells contain epitopes for the receptor. The protein is not detected in glomerular capillaries, in the glomerular mesangium, in the interstitium, or in blood vessels. Specific polyclonal antibodies directed against the 25(OH)D3 24-hydroxylase cytochrome P-450 demonstrate epitopes for the cytochrome in cells of the proximal tubule, the distal tubule, glomerular parietal epithelial cells, and mesangial cells. The protein is absent from interstitial cells. Calbindin D28k is present exclusively in principal cells of the distal tubule and collecting duct. In the human kidney, the VDR is present in cells where vitamin D-inducible proteins are found; conversely it is absent from cells where vitamin D-dependent proteins are not present.

scholarly journals Human kidney is a target for novel severe acute respiratory syndrome coronavirus 2 infection

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Bo Diao ◽  
Chenhui Wang ◽  
Rongshuai Wang ◽  
Zeqing Feng ◽  
Ji Zhang ◽  
...  
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Kidney Injury ◽  
Human Kidney ◽  
The Elderly ◽  
Tubular Damage ◽  
Clinical Parameters ◽  
Double Staining ◽  
Protein Deposits ◽  
Prostaglandin D Synthase

AbstractIt is unclear whether severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can directly infect human kidney, thus leading to acute kidney injury (AKI). Here, we perform a retrospective analysis of clinical parameters from 85 patients with laboratory-confirmed coronavirus disease 2019 (COVID-19); moreover, kidney histopathology from six additional COVID-19 patients with post-mortem examinations was performed. We find that 27% (23/85) of patients exhibited AKI. The elderly patients and cases with comorbidities (hypertension and heart failure) are more prone to develop AKI. Haematoxylin & eosin staining shows that the kidneys from COVID-19 autopsies have moderate to severe tubular damage. In situ hybridization assays illustrate that viral RNA accumulates in tubules. Immunohistochemistry shows nucleocapsid and spike protein deposits in the tubules, and immunofluorescence double staining shows that both antigens are restricted to the angiotensin converting enzyme-II-positive tubules. SARS-CoV-2 infection triggers the expression of hypoxic damage-associated molecules, including DP2 and prostaglandin D synthase in infected tubules. Moreover, it enhances CD68+ macrophages infiltration into the tubulointerstitium, and complement C5b-9 deposition on tubules is also observed. These results suggest that SARS-CoV-2 directly infects human kidney to mediate tubular pathogenesis and AKI.

scholarly journals Insulin-like growth factor binding protein 7 and tissue inhibitor of metalloproteinases-2: differential expression and secretion in human kidney tubule cells

2017 ◽  
Vol 312 (2) ◽  
pp. F284-F296 ◽  
Author(s):  
David R. Emlet ◽  
Nuria Pastor-Soler ◽  
Allison Marciszyn ◽  
Xiaoyan Wen ◽  
Hernando Gomez ◽  
...  
Keyword(s):  
Cell Culture ◽  
Acute Kidney Injury ◽  
Kidney Injury ◽  
Human Kidney ◽  
Distal Tubule ◽  
Cell Types ◽  
Tissue Inhibitor Of Metalloproteinases ◽  
Tubule Cell ◽  
Tubule Cells

We have characterized the expression and secretion of the acute kidney injury (AKI) biomarkers insulin-like growth factor binding protein 7 (IGFBP7) and tissue inhibitor of metalloproteinases-2 (TIMP-2) in human kidney epithelial cells in primary cell culture and tissue. We established cell culture model systems of primary kidney cells of proximal and distal tubule origin and observed that both proteins are indeed expressed and secreted in both tubule cell types in vitro. However, TIMP-2 is both expressed and secreted preferentially by cells of distal tubule origin, while IGFBP7 is equally expressed across tubule cell types yet preferentially secreted by cells of proximal tubule origin. In human kidney tissue, strong staining of IGFBP7 was seen in the luminal brush-border region of a subset of proximal tubule cells, and TIMP-2 stained intracellularly in distal tubules. Additionally, while some tubular colocalization of both biomarkers was identified with the injury markers kidney injury molecule-1 and neutrophil gelatinase-associated lipocalin, both biomarkers could also be seen alone, suggesting the possibility for differential mechanistic and/or temporal profiles of regulation of these early AKI biomarkers from known markers of injury. Last, an in vitro model of ischemia-reperfusion demonstrated enhancement of secretion of both markers early after reperfusion. This work provides a rationale for further investigation of these markers for their potential role in the pathogenesis of acute kidney injury.

Human kidney development

2015 ◽  
Author(s):  
Paul Winyard
Keyword(s):  
Kidney Development ◽  
Hormone Secretion ◽  
Human Kidney ◽  
Distal Tubule ◽  
Genetic Syndromes ◽  
Acid Base Balance ◽  
Waste Products ◽  
Molecular Control ◽  
Mesonephric Duct ◽  
Base Balance

The kidneys perform diverse functions including excretion of nitrogenous waste products, homeostasis of water, electrolytes and acid–base balance, and hormone secretion. The simplest functional unit within the kidneys is the nephron, which consists of specialized segments from glomerulus, through proximal tubule, loop of Henle, and distal tubule. Human nephrogenesis starts with two stages of transient kidneys, termed the pronephros and mesonephros, and ends with development of a permanent organ from the metanephros on each side. The latter consists of just a few hundred cells when it is formed in the fifth week of pregnancy but progresses to a nephron endowment of between 0.6 to 1.3 million by the time nephrogenesis is completed at 32–36 weeks of gestation. Key events during this process include outgrowth of the epithelial ureteric bud from the mesonephric duct, interactions between the bud and the metanephric blastema (a specific region of mesenchyme) that cause the bud to branch and mesenchyme to condense, epithelialization of the mesenchyme to form proximal parts of the nephron, and differentiation of segment specific cells. Molecular control of these events is being unpicked with data from human genetic syndromes and animal models, and this chapter highlights several of the most important factors/systems involved. Increased understanding of development is not just relevant to congenital kidney malformations, but may also be important in designing rational therapies for diseases of the mature kidney where recapitulation of developmental pathways is common.

Characterization of two potential immunotherapeutics in tumor infiltrating lymphocytes (TILs) using flow cytometry and Meso Scale Discovery.

2018 ◽  
Vol 36 (15_suppl) ◽  
pp. e15052-e15052
Author(s):  
Karen K Yam ◽  
Stephen Parker ◽  
Michael Agrez ◽  
Christopher Warburton ◽  
Adriana Alcantara ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Tumor Infiltrating Lymphocytes ◽  
Meso Scale Discovery ◽  
Infiltrating Lymphocytes ◽  
Meso Scale

scholarly journals Alpha-Hemoglobin-Stabilizing Protein: An Erythroid Molecular Chaperone

2011 ◽  
Vol 2011 ◽  
pp. 1-7 ◽  
Author(s):  
Maria Emília Favero ◽  
Fernando Ferreira Costa
Keyword(s):  
Molecular Chaperone ◽  
Specific Protein ◽  
Erythroid Cell ◽  
Ineffective Erythropoiesis ◽  
Cytotoxic Effects ◽  
Erythroid Precursor ◽  
Pathological Conditions ◽  
Alpha Globin ◽  
Health And Disease ◽  
And Function

Alpha-hemoglobin-stabilizing protein (AHSP) is an erythroid-specific protein that acts as a molecular chaperone for the free α chains of hemoglobin. Evidence strongly suggests that AHSP participates in hemoglobin synthesis and may act to neutralize the cytotoxic effects of excess free alpha-globin subunits that accumulate both in normal and beta-thalassemic erythroid precursor cells. As such, AHSP seems to be essential for normal erythropoiesis, and impaired upregulation of AHSP may lead to premature erythroid cell death, resulting in ineffective erythropoiesis. ReducedAHSPmRNA expression has been associated with clinical variability in some cases of β-thalassemia. It has been shown that αHb variants may also impair AHSP-αHb interactions, leading to pathological conditions that resemble α-thalassemia syndromes. The aim of this paper is to summarize current information concerning the structure and function of AHSP, focusing on its role in normal erythropoiesis and its relevance in health and disease.

Cloning and localization of a double-pore K channel, KCNK1: exclusive expression in distal nephron segments

1997 ◽  
Vol 273 (4) ◽  
pp. F663-F666 ◽  
Author(s):  
Marcelo Orias ◽  
Heino Velázquez ◽  
Freeman Tung ◽  
George Lee ◽  
Gary V. Desir
Keyword(s):  
Amino Acid ◽  
Collecting Duct ◽  
Human Kidney ◽  
Distal Tubule ◽  
Amino Acid Identity ◽  
Northern Analysis ◽  
K Channel ◽  
Thick Ascending Limb ◽  
Cortical Collecting Duct ◽  
K Channels

The K-selective channel, TOK1, recently identified in yeast, displays the unusual structural feature of having two putative pore regions, in contrast to all previously cloned K channels. Using the TOK1 pore regions as probes, we identified a human kidney cDNA encoding a 337-amino acid protein (hKCNK1) with four transmembrane segments and two pore regions containing the signature sequence of K channels. Amino acid identity to TOK1 is only 15% overall but 40% at the pores. Northern analysis indicates high expression of a 1.9-kb message in brain > kidney >> heart. Nephron segment localization, carried out in rabbit by reverse transcription-polymerase chain reaction, reveals that KCNK1 is expressed in cortical thick ascending limb, connecting tubule, and cortical collecting duct. It was not detected in the proximal tubule, medullary thick ascending limb, distal convoluted tubule, and glomerulus. We conclude that KCNK1 is a unique, double-pore, mammalian K channel, distantly related to the yeast channel TOK1, that is expressed in distal tubule and is a candidate to participate in renal K homeostasis.

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