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The different molecular forms of urine neutrophil gelatinase-associated lipocalin present in dogs with urinary diseases.

Hsu WL, Chiou HC, Tung KC, Belot G, Virilli A, Wong ML, Lin FY, Lee YJ - BMC Vet. Res. (2014)

Bottom Line: In particular, monomer was detected more frequently in patients with renal disease than those with non-renal diseases; while the dimer form appeared in a significantly higher percentage of cases with pyuria compared to those without pyuria.The NGAL/MMP-9 complex was found to exist not only in the patients with cystitis, but also in the cases with renal injury.Different molecular forms of uNGAL can indicate different origins of the urinary abnormalities.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Veterinary Clinical Science, School of Veterinary Medicine, College of Bio-Resources and Agriculture, National Taiwan University, No,1, Sec,4, Roosevelt Rd, Taipei, Taiwan. yajanelee@ntu.edu.tw.

ABSTRACT

Background: Neutrophil gelatinase-associated lipocalin (NGAL) is a useful biomarker for the early prediction of renal diseases. NGAL may exist as monomer, dimer and/or NGAL/MMP-9 complex forms in humans. In this study, the existence of various forms of NGAL in urine (uNGAL) was determined and whether these forms are related to the different urinary diseases found in dogs is further discussed.

Results: Eighty-one urine samples from dogs with different forms of renal disease (41), pyuria (19) and a number of non-renal related diseases (10), as well as healthy dogs (11), were collected. uNGAL concentrations and their molecular forms in dogs were measured by ELISA and Western blot analysis, respectively. The uNGAL concentrations of dogs with pyuria (median: 15.35 ng/mL) were significantly higher than those of the healthy control animals (median: 3.92 ng/mL) (p < 0.01), but lower than those of dogs with renal diseases (median: 23.77 ng/mL). Each NGAL molecular form could be detected in dog urine. In particular, monomer was detected more frequently in patients with renal disease than those with non-renal diseases; while the dimer form appeared in a significantly higher percentage of cases with pyuria compared to those without pyuria. The NGAL/MMP-9 complex was found to exist not only in the patients with cystitis, but also in the cases with renal injury.

Conclusion: Different molecular forms of uNGAL can indicate different origins of the urinary abnormalities. Determining the molecular forms of uNGAL present in diseased dogs may provide clinical workers with a tool that will help the early and more precise detection of different urinary diseases.

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Related in: MedlinePlus

Western blot analysis of the various NGAL molecular forms. Urine samples representing the PYU (lane 1, 5), AKI (lane 2), non-RP (enteritis, lane 3), CKD (lane 4), and control (lane 6) groups were resolved using non-reducing SDS-PAGE followed by Western blot analysis using polyclonal antibodies against canine NGAL (A), or canine MMP-9 (B). Three different NGAL related proteins, as indicated by the arrowheads, were recognized by the anti-NGAL antibodies; the protein with highest molecular weight could also be detected using the anti-MMP-9 antibodies. M, D, and C indicate the monomeric NGAL, the dimeric NGAL, and the NGAL/MMP-9 complex, respectively.
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Figure 3: Western blot analysis of the various NGAL molecular forms. Urine samples representing the PYU (lane 1, 5), AKI (lane 2), non-RP (enteritis, lane 3), CKD (lane 4), and control (lane 6) groups were resolved using non-reducing SDS-PAGE followed by Western blot analysis using polyclonal antibodies against canine NGAL (A), or canine MMP-9 (B). Three different NGAL related proteins, as indicated by the arrowheads, were recognized by the anti-NGAL antibodies; the protein with highest molecular weight could also be detected using the anti-MMP-9 antibodies. M, D, and C indicate the monomeric NGAL, the dimeric NGAL, and the NGAL/MMP-9 complex, respectively.

Mentions: Canine MMP9 antibody was successfully produced (Figure 2). As shown in Figure 3A, three different forms of uNGAL were successfully detected in all the urine samples collected from patients (Figure 3). The two forms with smaller molecular weight, namely ~25 kDa and ~50 kDa, represent the monomeric and dimeric forms of NGAL, respectively. The remaining higher molecular weight form of the NGAL related protein, with a molecular weight over 130 kDa, was then identified as the NGAL/MMP-9 complex by Western blot analysis using antibodies against canine-MMP-9 (Figure 3B).


The different molecular forms of urine neutrophil gelatinase-associated lipocalin present in dogs with urinary diseases.

Hsu WL, Chiou HC, Tung KC, Belot G, Virilli A, Wong ML, Lin FY, Lee YJ - BMC Vet. Res. (2014)

Western blot analysis of the various NGAL molecular forms. Urine samples representing the PYU (lane 1, 5), AKI (lane 2), non-RP (enteritis, lane 3), CKD (lane 4), and control (lane 6) groups were resolved using non-reducing SDS-PAGE followed by Western blot analysis using polyclonal antibodies against canine NGAL (A), or canine MMP-9 (B). Three different NGAL related proteins, as indicated by the arrowheads, were recognized by the anti-NGAL antibodies; the protein with highest molecular weight could also be detected using the anti-MMP-9 antibodies. M, D, and C indicate the monomeric NGAL, the dimeric NGAL, and the NGAL/MMP-9 complex, respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4236819&req=5

Figure 3: Western blot analysis of the various NGAL molecular forms. Urine samples representing the PYU (lane 1, 5), AKI (lane 2), non-RP (enteritis, lane 3), CKD (lane 4), and control (lane 6) groups were resolved using non-reducing SDS-PAGE followed by Western blot analysis using polyclonal antibodies against canine NGAL (A), or canine MMP-9 (B). Three different NGAL related proteins, as indicated by the arrowheads, were recognized by the anti-NGAL antibodies; the protein with highest molecular weight could also be detected using the anti-MMP-9 antibodies. M, D, and C indicate the monomeric NGAL, the dimeric NGAL, and the NGAL/MMP-9 complex, respectively.
Mentions: Canine MMP9 antibody was successfully produced (Figure 2). As shown in Figure 3A, three different forms of uNGAL were successfully detected in all the urine samples collected from patients (Figure 3). The two forms with smaller molecular weight, namely ~25 kDa and ~50 kDa, represent the monomeric and dimeric forms of NGAL, respectively. The remaining higher molecular weight form of the NGAL related protein, with a molecular weight over 130 kDa, was then identified as the NGAL/MMP-9 complex by Western blot analysis using antibodies against canine-MMP-9 (Figure 3B).

Bottom Line: In particular, monomer was detected more frequently in patients with renal disease than those with non-renal diseases; while the dimer form appeared in a significantly higher percentage of cases with pyuria compared to those without pyuria.The NGAL/MMP-9 complex was found to exist not only in the patients with cystitis, but also in the cases with renal injury.Different molecular forms of uNGAL can indicate different origins of the urinary abnormalities.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Veterinary Clinical Science, School of Veterinary Medicine, College of Bio-Resources and Agriculture, National Taiwan University, No,1, Sec,4, Roosevelt Rd, Taipei, Taiwan. yajanelee@ntu.edu.tw.

ABSTRACT

Background: Neutrophil gelatinase-associated lipocalin (NGAL) is a useful biomarker for the early prediction of renal diseases. NGAL may exist as monomer, dimer and/or NGAL/MMP-9 complex forms in humans. In this study, the existence of various forms of NGAL in urine (uNGAL) was determined and whether these forms are related to the different urinary diseases found in dogs is further discussed.

Results: Eighty-one urine samples from dogs with different forms of renal disease (41), pyuria (19) and a number of non-renal related diseases (10), as well as healthy dogs (11), were collected. uNGAL concentrations and their molecular forms in dogs were measured by ELISA and Western blot analysis, respectively. The uNGAL concentrations of dogs with pyuria (median: 15.35 ng/mL) were significantly higher than those of the healthy control animals (median: 3.92 ng/mL) (p < 0.01), but lower than those of dogs with renal diseases (median: 23.77 ng/mL). Each NGAL molecular form could be detected in dog urine. In particular, monomer was detected more frequently in patients with renal disease than those with non-renal diseases; while the dimer form appeared in a significantly higher percentage of cases with pyuria compared to those without pyuria. The NGAL/MMP-9 complex was found to exist not only in the patients with cystitis, but also in the cases with renal injury.

Conclusion: Different molecular forms of uNGAL can indicate different origins of the urinary abnormalities. Determining the molecular forms of uNGAL present in diseased dogs may provide clinical workers with a tool that will help the early and more precise detection of different urinary diseases.

Show MeSH
Related in: MedlinePlus