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PloS one2020; 15(10); e0240586; doi: 10.1371/journal.pone.0240586

Evaluation of podocin in urine in horses using qualitative and quantitative methods.

Abstract: No sensitive method for diagnosing early kidney dysfunction in horses has been identified so far. Many studies carried out in humans and small animals show that podocin can be useful to diagnose various kidney diseases, mainly affecting the glomeruli. The aim of this study was to perform a qualitative and quantitative analysis of podocin in urine samples obtained from healthy horses, horses with clinical kidney dysfunction and horses at risk of acute kidney injury. The study objectives aimed to assess: (1) whether the selected podocin tryptic peptide for LC-MS-MRM allows for podocin detection in horse; and (2) whether the species-specific ELISA test makes this detection possible as well;, (3) whether the chosen methods are sensitive enough to detect kidney dysfunction and glomerular injury, (4) whether the results of the tests applying both methods correspond with one another, (5) whether the results correlate with the hematological and biochemical data. The signals that may indicate the presence of trypsin fragments of podocin were found in three healthy horses, all the horses diagnosed with kidney dysfunction and half of the animals at risk for acute kidney injury. The concentration of podocin, diagnosed with the ELISA test was as follows: from 0.19 to 1.2 ng/ml in healthy animals, from 0.19 to 20.0 ng/ml in AKI horses, from 0.29 to 5.71 ng/ml in horses at risk for acute kidney injury. The results of both methods corresponded significantly. Podocin may be a potential biomarker of clinical kidney disease in horses and may be used in the detection of glomerular injury. However, its use is limited by the possibility of physiological podocyturia. LC-MS-MRM seems to be a more sensitive method to evaluate the presence of podocin than the ELISA test, whilst selected tryptic peptides of podocin appear to apply to horses. The ELISA test showed greater effectiveness in excluding the disease than in confirming it.
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Publication Date: 2020-10-15 PubMed ID: 33057359PubMed Central: PMC7561189DOI: 10.1371/journal.pone.0240586Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article presents an innovative evaluation of podocin levels in horse urine as a marker for kidney disease and injury, using Liquid Chromatography-Mass Spectrometry Multiple Reaction Monitoring (LC-MS-MRM) and Enzyme-Linked Immunosorbent Assay (ELISA) methods. Briefly, the study discovered signals of podocin, a potential biomarker for horse kidney dysfunction, applying both methods — with LC-MS-MRM appearing more sensitive than the ELISA test.

Objectives and Methods

  • The research aimed to establish a reliable way to identify early kidney dysfunction in horses, specifically by conducting a qualitative and quantitative analysis of podocin in their urine. Podocin, identified in human and small animal kidney disease studies, mostly affects the kidneys’ filtration units: the glomeruli.
  • The researchers created markers of success that included identifying if a selected podocin peptide could be used for LC-MS-MRM detection, finding out if a species-specific ELISA test was applicable as well, and understanding whether both methods had enough sensitivity to detect kidney dysfunction and glomerular injury in horses.
  • They also examined the correlation between both methods’ results and compared them with hematological and biochemical data for validation. In this context, the ELISA test was expected to be more effective in ruling out disease than in confirming it.

Results and Outcomes

  • The study yielded signals that potentially indicated the presence of podocin in three healthy horses, all horses with diagnosed kidney dysfunction, and half the horses at risk for acute kidney injury.
  • The detected podocin concentration using ELISA testing ranged between 0.19 and 1.2 ng/ml in healthy horses, 0.19 to 20.0 ng/ml in horses with Acute Kidney Injury (AKI), and 0.29 to 5.71 ng/ml in horses at risk for AKI.
  • The outcomes of both methods showed a strong correspondence, suggesting that podocin might be a potential biomarker for clinical kidney disease in horses and could be used to detect glomerular injury.
  • However, the use of podocin as a marker is complicated by the potential of physiological podocyturia. This means that the presence of podocytes (cells from the kidney that may show up due to normal physiological processes) in the urine could skew the results.
  • Of the two methods employed, LC-MS-MRM appeared to be more sensitive at evaluating the presence of podocin than ELISA. The ELISA test, on the contrary, showed greater effectiveness in excluding the disease than in confirming it.
  • The study’s comprehensive findings make a case for the preliminary applicability of podocin tryptic peptides in horses, though the physiological appearance of podocytes in urine necessitates further investigation for its evolution as a valid diagnostic tool.

Cite This Article

APA
Siwińska N, Pasławska U, Bąchor R, Szczepankiewicz B, Żak A, Grocholska P, Szewczuk Z. (2020). Evaluation of podocin in urine in horses using qualitative and quantitative methods. PLoS One, 15(10), e0240586. https://doi.org/10.1371/journal.pone.0240586

Publication

PloS one
ISSN: 1932-6203
NlmUniqueID: 101285081
Country: United States
Language: English
Volume: 15
Issue: 10
Pages: e0240586
PII: e0240586

Researcher Affiliations

Siwińska, Natalia
  • Department of Internal Diseases with Clinic for Horses, Dogs and Cats, Faculty of Veterinary Medicine, Wrocław University of Environmental and Life Sciences, Wrocław, Poland.
Pasławska, Urszula
  • Department of Internal Diseases with Clinic for Horses, Dogs and Cats, Faculty of Veterinary Medicine, Wrocław University of Environmental and Life Sciences, Wrocław, Poland.
  • Veterinary Institute, Faculty of Biological and Veterinary Sciences, Nicolaus Copernicus University, Toruń, Poland.
Bąchor, Remigiusz
  • Faculty of Chemistry, University of Wrocław, Wrocław, Poland.
Szczepankiewicz, Barbara
  • Department of Internal Diseases with Clinic for Horses, Dogs and Cats, Faculty of Veterinary Medicine, Wrocław University of Environmental and Life Sciences, Wrocław, Poland.
Żak, Agnieszka
  • Department of Internal Diseases with Clinic for Horses, Dogs and Cats, Faculty of Veterinary Medicine, Wrocław University of Environmental and Life Sciences, Wrocław, Poland.
Grocholska, Paulina
  • Faculty of Chemistry, University of Wrocław, Wrocław, Poland.
Szewczuk, Zbigniew
  • Faculty of Chemistry, University of Wrocław, Wrocław, Poland.

MeSH Terms

  • Acute Kidney Injury / diagnosis
  • Acute Kidney Injury / pathology
  • Acute Kidney Injury / urine
  • Acute Kidney Injury / veterinary
  • Animals
  • Biomarkers / urine
  • Female
  • Glomerulonephritis / diagnosis
  • Glomerulonephritis / pathology
  • Glomerulonephritis / urine
  • Glomerulonephritis / veterinary
  • Horses / urine
  • Intracellular Signaling Peptides and Proteins / metabolism
  • Intracellular Signaling Peptides and Proteins / urine
  • Male
  • Membrane Proteins / metabolism
  • Membrane Proteins / urine
  • Podocytes / metabolism
  • Podocytes / pathology
  • Prognosis

Conflict of Interest Statement

The authors have declared that no competing interests exist.

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