Search

The aims of the current study were to assess whether sodium dodecyl sulphate-agarose gel electrophoresis (SDS-AGE) and high-resolution electrophoresis (HRE) can identify dogs with a urinary protein-to-creatinine ratio (UPC ratio) >0.2 and whether HRE can provide preliminary information about the type of proteinuria, using SDS-AGE as a reference method. HRE and SDS-AGE were conducted on 87 urine samples classified according to the International Renal Interest Society as non-proteinuric (NP; UPC ratio: <0.20; 32/87), borderline proteinuric (BP; UPC ratio: 0.21-0.50; 15/87), or proteinuric (P; UPC ratio: >0.51; 40/87). SDS-AGE and HRE were positive in 14 out of 32 and 3 out of 32 NP samples and in 52 out of 55 and 40 out of 55 samples with a UPC ratio >0.20, respectively. The concordance between HRE or SDS and UPC ratio was comparable (κ = 0.59; κ = 0.55). However, specificity (90%) and positive likelihood ratio (7.76) were higher for HRE than for SDS-AGE (56% and 2.16) while sensitivity was lower (73% vs. 94%). The analysis of HRE results revealed that a percentage of albumin >41.4% and an albumin/α(1)-globulin ratio (alb/α(1) ratio) >1.46 can identify samples classified by SDS-AGE as affected by glomerular proteinuria while a percentage of α(1)-globulin >40.8% and an alb/α(1) ratio <0.84 can identify samples classified by SDS-AGE as affected by tubular proteinuria. In conclusion, both SDS-AGE and HRE could misclassify samples with a UPC ratio higher or lower than 0.20. Therefore, UPC ratio must always be determined before conducting these tests. The percentage of albumin and α(1)-globulin or the alb/α(1) ratio determined by HRE can provide preliminary information about the origin of proteinuria.

Objective: To assess recovery of renal parenchymal thickness and urinary protein levels in patients with severely hydronephrotic kidneys after nephrostomy placement., Methods: Fourteen patients (median age 1 year, range 6 months-7 years) who underwent nephrostomy placement for unilateral ureteropelvic junction obstruction at our hospital between May 2007 and January 2009 were included in a retrospective analysis. All patients had severe hydronephrosis, with a median parenchymal thickness of 1.8 mm (range 1-2.5 mm). Kidney morphology was examined by ultrasound before the procedure and 1, 2, 3, 4, 6 and 8 weeks after. Urinary proteins (including albumin, immunoglobulin [IgG], alpha2-macroglobulin, alpha1-microglobulin, beta2-microglobulin [beta2-MG] and kappa chain) and creatinine levels were also tested during these follow-up visits. Fifteen healthy children were assessed for urinary protein levels as well and made up the control group., Results: Parenchymal thickness increased within 4 weeks of nephrostomy placement. Kidney volumes were significantly decreased within 2 weeks. No further changes in morphology were detected after 4 weeks. Urinary alpha1-microglobulin and beta2-MG levels decreased to baseline within 1 and 4 weeks, respectively. Urinary albumin, IgG, alpha2-macroglobulin and kappa chain levels decreased gradually after nephrostomy, but did not return to baseline within 8 weeks., Conclusions: After nephrostomy placement, parenchymal thickness increases within 4 weeks, tubular function returns to normal earlier than glomerular function and glomerular membrane repair is inversely correlated with the severity of damage.

The electrophoretic pattern of concentrated urine samples can be used to identify the type of proteins leaking into the urine and has diagnostic and prognostic value, providing information about the location (glomerular or tubular) and degree of renal injury. This test usually requires a 24-hour urine collection, which can be inconvenient because of its heavy dependence on patient compliance and frequently is unreliable because of errors in collecting a complete 24-hour urine sample. In this study, we compared the electrophoretic pattern in 24-hour urine collections and random samples among patients with glomerular diseases and a wide range of proteinuria. Forty adult patients were evaluated; 24-hour urine collections and random urine samples were analyzed. Protein concentrations were determined using the sulfosalicylic acid method standardized with human serum. Electrophoresis was performed with concentrated urine samples (Ultrafree, PF/Millipore Corporation, Bedford, MA) using Beckman Paragon Electrophoresis System (agarose gels and blue staining; Beckman Instruments, Inc, Brea, CA). Densitometric scanning of electrophoretic pattern (Appraise Clinical Densitometer; Beckman Instruments, Inc) was performed, and the results were reported in percentages of each observed fraction. Our results revealed that despite the significant difference between protein concentration in 24-hour collections and in random samples, the pattern of protein excretion, in percentage basis, remains the same. There were no differences between the albumin, alpha(1)-globulin, alpha(2)-globulin, beta-globulin, and gamma-globulin fractions in both types of specimens. This study shows that, at least in glomerular proteinuria, the electrophoretic analysis of the urine can be performed accurately in random samples, avoiding the inconveniences and errors of a 24-hour urine collection., (Copyright 2002 by the National Kidney Foundation, Inc.)

The protein fractions in urine from proteinuric dogs with and without pyoderma were estimated. Fifteen dogs with pyoderma (five with superficial and 10 with deep pyoderma) were compared with 10 dogs with glomerulopathy and 27 dogs with diseases other than pyoderma or urinary tract problems. Agarose gel electrophoresis was used to fractionate the proteins. Three types of electrophoretogram were obtained with albuminuria, globulinuria and serum-like profiles. An albuminuria profile was found in eight of the 27 dogs with other diseases, in three of the five dogs with superficial pyoderma, in eight of the 10 dogs with deep pyoderma and in all 10 dogs with glomerulopathy. The albuminuria profile (mean [sem] albumin/globulin ratio 1.98 [0.10]) was also characterised by alpha 1b, alpha 2a and beta 2 globulin peaks in all 29 dogs with this profile, which was therefore thought to indicate that albuminuria (glomerular proteinuria) was a result of glomerular damage and inflammation because alpha 1b, alpha 2a, and beta 2 globulins are considered to be acute phase proteins. The serum-like profile (mean [sem] albumin/globulin ratio 0.72 [0.01]) was observed in 13 per cent of the proteinuric dogs examined and contained all the protein fractions normally detected by electrophoresis of serum. The profile was considered to be a variant from of the albuminuria profile, probably indicating advanced glomerular lesions and inflammation. The globulinuria profile (mean [sem] albumin/globulin ratio 0.33 [0.08]) was significantly different from the other two in that it was characterised by a low albumin peak and the presence of globulin fractions not clearly distinguishable from each other because of their confluency and absence of individual peaks. This profile could indicate severe glomerulotubular lesions and degradation of certain protein fractions. It could also be a result of increased secretion of tissue and other proteins by damaged tubules. It was concluded that glomerular damage leads to glomerular proteinuria characterised by high proportions of albumin together with alpha 1b, alpha 2a and beta 2 globulins in lower but significantly diagnostic proportions.

We fractionated normal urinary proteins obtained from 40 healthy subjects using cellulose acetate membrane electrophoresis and stained them with Acid Violet 17. The electrophoretic patterns were classified into four groups. Each of groups I, II, III, IV had an albumin peak and 1, 2, 3, and 4 additional globulin peaks, respectively. Within-day variation study showed that the pattern was fundamentally specific to the individual, although some intermediate cases were observed. We were unable to determine which type was standard for normal subjects. However, the concentration of Tamm-Horsfall protein was speculated to be an important factor in determining the patterns. Group III showed significantly higher values than group I in urine albumin, total protein, and beta-N-acetyl-D-glucosaminidase and this group was believed to include subjects in the subclinical stage of a glomerular disease. All specimens belonging to group IV showed an obvious fraction of alpha 1 globulin which is often found in urine specimens of patients with renal diseases of tubular origin or other pathological conditions.

Because standard urinary protein test paper reacts more strongly to albumin than to globulin, we attempted to develop a test paper sensitive to globulin by applying chromatic analysis principles used in quantitative analysis of urinary protein. We developed a new urinary protein test paper (URINE-TP) containing a reagent of ACID VIOLET 17. It was found that URINE-TP reacts with the same strength to gamma-globulin as to albumin. This new test paper therefore allows accurate detection of Bence Jones protein, which was formerly detectable with standard test paper. There was a correlation between values for urinary protein obtained with URINE-TP and those obtained by quantitative analysis. As with standard test paper, URINE-TP indicates urinary protein values in the normal range by a lack of reaction, but on URINE-TP positive and negative results are more clearly distinguishable than on standard test paper. From these results, we conclude that URINE-TP enables detection of both gamma-globulin and albumin in the urine, and that is as sensitive as the analytical method in the screening of urinary proteins.

A precipitating reagent comprising partially neutralized trichloroacetic acid with magnesium sulfate, sodium dodecyl sulfate and polyethylene glycol is used for automatic determination of proteins from urine by centrifugal analysis and continuous flow analysis. We compare and discuss the results of both methods, their limits of validity and incorporation in an automated laboratory.

Two commonly used brands of reagent strip (dipsticks) were evlauated for their sensitivity to Bence-Jones and seven other urinary proteins. Both brands showed significant differences in sensitivity to albumin, glycoprotein, ribonuclease and lysozyme; both were most sensitive to albumin and least sensitive to globulin. Furthermore, their comparative sensitivities to these proteins also differed markedly. These differences in sensitivity could lead to underestimation of protein content in urine specimens. Tests on urines from patients with multiple myeloma showed that a negative urinary dipstick test result did not rule out the presence of the disease.

The authors propose a convenient method for the separation of low molecular weight proteins, present in tubular urines. In particular, the method is used for the purification of beta2 microglobulin, after one recycling, and also for the isolation of the various post-gamma globulins which can later be separated on DEAE Sephadex.

High resolution electrophoresis was used to evaluate protein excretion patterns in six cadmium-exposed individuals with proteinuria, seven subjects with nonspecific nephropathies, and four normal unexposed subjects. The aim of the investigation was to determine (a) the type of excretion pattern (i.e., glomerular, tubular, or mixed) associated with cadmium exposure and (b) if the pattern in the cadmium-exposed individuals was distinctly different from subjects with nonspecific nephropathies. The electrophoretic results were consistent with the quantitative results for the cadmium-exposed workers. The results suggest that the pattern associated with cadmium exposure can be glomerular or mixed and that it is different (i.e., no gamma band) from nonspecific nephropathies.

Administration of a single oral dose of 60 mg/kg O,O,S-trimethyl phosphorothioate (OOS-Me), a malathion impurity, resulted in a substantial increase in the amounts of amino acids along with a change in the nature of proteins excreted in the urine of treated rats. In contrast to control rats, a small increase in albumin and a small decrease in alpha 1-globulin were observed. However, alpha2-, beta- and gamma 1-globulin, which were not detected in the urine of control rats, were found in substantial amounts in the urine of OOS-Me-treated rats. These findings, coupled with observed increases in urinary glucose levels and consistent specific gravity readings of 1.01 even though treated rats were experiencing oliguria, provide evidence for OOS-Me-induced kidney tubule damage.

The urinary total protein excretion was determined in 270, 18-24 hr urine samples from 130 healthy children of different age groups using the tannic acid-Fe3+-method of Yatzidis [1977]. The daily protein excretion of premature infants in the first month of life varies between 14-60 mg, with a mean of 29 mg, and that of fullterm newborn infants between 15-68 mg, with a mean of 32 mg. Protein excretion increases with age and amounts to 29-238 mg (mean 83 mg) in 10-16 year old children. Thus, the urinary protein concentration during the neonatal period is high when compared to adult values. This explains the "trace" and "positive" reactions frequently obtained in this period of life with Albustix. In 92 urine samples proteins were fractionated by sodium dodecyl sulphate gel discelectrophoresis. Hemoglobinuria was found during the first weeks of life and tubular type proteinuria was found in newborns and infants. The present data suggest that the proteinuria is due to ineffective proximal tubular reabsorption of low molecular weight microproteins as a result of glomerulo-tubular imblance in early life.

Automatic assay of urinary total proteins on a centrifugal analyser. The assay of urinary proteins is achieved opacimetrically after precipitation by trichloracetic acid. The addition of tensio-active agents lates the initiation of precipitation, helps the formation of a microprecipitate thus hold in suspension. The technique is linear from 0 to 2,5 g/l and for every albumin/globulins ratio between pure albumin and pure globulin.