alcian-blue and Proteinuria

Some preliminary observations suggest that predisposition to a particular type of glomerulonephritis (GN) may be connected with the genetically determined charge of the glomerular capillary wall. A correlation between erythrocyte surface and the glomerular capillary wall charges has also been observed. The purpose of this study was to verify and extend previous investigations. Therefore we measured erythrocyte and platelet surface charge from patients with idiopathic membranous and mesangial GN as well as idiopathic membranoproliferative GN and lupus nephritis.. The erythrocyte and platelet surface charge was determined by the binding of the cationic dye, alcian blue (AB). A fresh alcoholic AB solution was made for each experiment, which were run in batches of four, each including cells from a healthy person and from patients each with a different type of GN.. In patients with idiopathic membranous and membranoproliferative GN, a significant decrease in the erythrocyte and platelet charges was observed irrespective of their clinical state (remission or nephrotic syndrome). Erythrocyte charge was decreased despite the normal amount of membranous sialic acid. In contrast, patients with idiopathic mesangial GN, in complete or partial remission, exhibited normal erythrocyte and platelet surface charges. Exclusively in this type of GN, the appearance of nephrotic proteinuria was associated with a slight decrease, the erythrocyte charge, which was not statistically significant (P > 0.1). A reduction in the negative erythrocyte charge in lupus nephritis was less in magnitude than in idiopathic membranous or membranoproliferative GN, and occurred independently of the level of daily proteinuria, whereas the platelet charge was normal.. The decrease of the erythrocyte and platelet charge in idiopathic membranous and mebranoproliferative GN seems to be a pre-morbid feature.

Topics: Adolescent; Adult; Aged; Alcian Blue; Blood Platelets; Case-Control Studies; Cell Membrane; Coloring Agents; Electrochemistry; Erythrocyte Membrane; Female; Glomerulonephritis; Glomerulonephritis, Membranoproliferative; Glomerulonephritis, Membranous; Humans; Lupus Nephritis; Male; Membrane Potentials; Middle Aged; Nephrotic Syndrome; Proteinuria; Surface Properties

We studied alcian blue (AB) binding to red blood cells (RBC) [ABRBC] in 89 children and adults with a variety of glomerular diseases. ABRBC was significantly reduced in the group as a whole when compared with healthy controls (P < 0.001). A moderate correlation between the degree of proteinuria and ABRBC was detected in the children with renal disease (r = 0.43, P < 0.001), but not in adult patients. A significant reduction in ABRBC was detected in 30 of the above children who had Steroid Responsive Nephrotic Syndrome (MCNS) compared to their controls (40.57 +/- 16.6 v. 75.09 +/- 7.81, P = 0.001). A similar decrease from normal values was observed in other childhood glomerular diseases: focal glomerulosclerosis, post-streptococcal glomerulonephritis, membranous nephropathy (52.31 +/- 26.07 v. 79.44 +/- 5.88, P = 0.001). Mean ABRBC was lower in MCNS than other histological groups (P = 0.0518). ABRBC was age-dependent (r = -0.399, P < 0.02). These findings provide further indirect evidence of the relative roles of charge and size selective filters in glomerular basement membrane (GBM) in renal diseases, and suggest that depletion of anionic charge may be a major cause of proteinuria in MCNS.

Topics: Adolescent; Adult; Alcian Blue; Analysis of Variance; Case-Control Studies; Child; Child, Preschool; Electric Conductivity; Erythrocytes; Female; Glomerulonephritis; Humans; Male; Proteinuria; Regression Analysis; Statistics, Nonparametric

We studied direct and indirect methods of measuring membrane charge by detecting fixed anionic sites with polyethylenimine (PEI) on the glomerular basement membrane (GBM) and Alcian blue on red blood cell (RBC) membrane (ABRBC), respectively, in 40 children with nephrotic syndrome (NS). Size selectivity of the GBM was measured indirectly by fine analysis of urinary proteins with sodium dodecyl sulfate-polyacrylamide gel electrophoresis in 22 of these children. Correlation between ABRBC and PEI was strongest (r = 0.79; p = 0.0037) in 11 children with steroid-responsive NS (SRNS), moderate (r = 0.31) in 10 children with focal glomerulosclerosis (FGS), and absent in 14 children with hepatitis B antigen membranous nephropathy (MGN) and 5 with mesangioproliferative glomerulonephritis (MPGN). ABRBC and PEI were reduced in the group as a whole as compared with their controls (ABRBC: 44.53 +/- 9.81 vs 71.54 +/- 12.14, p < 0.05; PEI: 16.31 +/- 4.34 vs 33.3 +/- 1.09, p < 0.005). This reduction was greater in SRNS (26.35 +/- 7.15 p = 0.004) but was also detected in the remainder of the group taken together (52.31 +/- 26.07, p < 0.001). Excretion of glomerular proteins was restricted by size (< or = 80 kd) in SRNS but unrestricted (< or = 80 kd plus > 80 kd) in FGS, MGN, and MPGN. The main cause of proteinuria is likely to be depletion of negative charge on the GBM in SRNS, and distortion of capillary pore size in MGN and MPGN, with probable overlap of these mechanisms in each disease, especially in FGS. Basement membrane injury appears widespread in SRNS but confined to the kidney in MGN and MPGN.

Topics: Alcian Blue; Basement Membrane; Biopsy; Child; Child, Preschool; Erythrocyte Membrane; Glomerulonephritis, Membranous; Glomerulosclerosis, Focal Segmental; Hepatitis B; Humans; In Vitro Techniques; Kidney Glomerulus; Nephrotic Syndrome; Proteinuria

Changes of milieu interieur in nephrotic syndrome (NS) have many consequences in various organs. We have measured the electrical charge of erythrocytes (Ery) with binding of alcian blue (AB) in 18 children with relapse of NS (12 minimal changes, 3 membranous and 3 mesangioproliferative glomerulonephritis) and 15 healthy children. The most important finding was that the binding of AB to Ery in patients with minimal change nephrotic syndrome (MCNS) and other aetiologies of NS was significantly less than that in the control group (p < 0.05). In addition, we have studied the thermal denaturation of the Ery membranes by differential scanning microcalorimetry. In some children with NS we have seen the splitting of B transition. We suppose that these phenomena occur as the result of structural change, which may involve lipoprotein components of the cytoskeletal network.

Topics: Alcian Blue; Calorimetry, Differential Scanning; Child, Preschool; Erythrocyte Membrane; Erythrocytes; Female; Humans; Male; Nephrotic Syndrome; Proteinuria

In workers exposed to Cd (8 years on the average), we have found a significant decrease of sialic acid in erythrocyte membranes (22.61 +/- 1.84 vs 25.80 +/- 3.01 micrograms/mg of protein in controls, p less than 0.05) and an increase of sialic acid concentration in both urine (276.7 +/- 132.3 vs 174.5 +/- 70.9 micrograms/g of creatinine, p less than 0.05) and plasma (761.8 +/- 83.5 vs 640.4 +/- 70.7 micrograms/ml, p less than 0.01). In rats exposed to Cd (100 ppm in drinking water for 5.5 months), we have observed a reduction of the sialic acid level in erythrocyte membranes (31.4 +/- 1.2 vs 33.4 +/- 1.1 micrograms/mg of protein, p less than 0.01) and glomeruli (12.5 +/- 1.3 vs 13.9 +/- 1.6 micrograms/mg of protein, p less than 0.05). These effects in Cd treated rats were accompanied by a loss of the glomerular barrier selectively as reflected by an increased urinary output of albumin and transferrin. After 10 months of Cd exposure, the albuminuria and transferrinuria were negatively correlated with the sialic acid content of glomerular membranes (r = -0.47 and -0.51, p less than 0.05), which suggests that the depletion of sialic acid is involved in the loss of glomerular barrier function induced by long term Cd exposure. In Cd-treated rats, sialidase activity was enhanced in kidney cortex and in serum but not in glomeruli.

Topics: Adult; Alcian Blue; Animals; Cadmium; Erythrocytes; Female; Humans; Kidney Cortex; Male; Middle Aged; N-Acetylneuraminic Acid; Occupational Exposure; Proteinuria; Rats; Rats, Inbred Strains; Sialic Acids

Topics: Alcian Blue; Animals; Erythrocytes; Humans; Indoles; Membrane Potentials; Nephrotic Syndrome; Proteinuria; Rabbits; Sheep

The negative charge on the surface of red blood cells may reflect the charge on the glomerular capillaries. Measurements were made of the charge on the red cells of patients with minimal change nephropathy (MCN), membranous nephropathy (IMN), and IgA nephropathy (IgAN) during complete or partial remission. The charge was significantly less in patients with MCN and IMN than in patients with IgAN. Since blood was taken during clinical remission, the differences probably represent the pre-morbid state. Low charge was associated with MCN and IMN, which usually present with heavy proteinuria, and high charge with IgA nephropathy, in which heavy proteinuria is rare. Patients with low red-cell charge are, therefore, susceptible to the development of heavy proteinuria and patients with high charge are protected. The site of deposition or formation of immune complexes may also be determined indirectly by charge.

Topics: Adolescent; Adult; Aged; Alcian Blue; Erythrocyte Membrane; Female; Glomerulonephritis; Glomerulonephritis, IGA; Humans; Immunoglobulin A; Ions; Male; Middle Aged; Nephrosis, Lipoid; Proteinuria; Sex Factors

The presence and distribution of anionic sites in the glomerular basement membrane and visceral epithelial cell coat has been demonstrated. No definite decrease in intensity or periodicity of staining of basement membrane particulate sites was seen in protein overload proteinuric animals and only one staining technique employed for electron microscopy (alcian blue 8GX) demonstrated a focal decrease in visceral epithelial cell coat staining in severely damaged glomeruli. A decrease in overall glomerular staining was also demonstrated by quantitative analysis of colloidal iron staining by light microscopy. The findings differ from those described in puromycin aminonucleoside nephropathy and nephrotoxic nephritis. Staining was demonstrated also in other basement membranes, in Bowman's capsule and along interstitial collagen fibres.

Topics: Alcian Blue; Animals; Anions; Basement Membrane; Binding Sites; Cell Membrane; Epithelium; Female; Iron; Kidney Glomerulus; Microscopy, Electron; Polyethyleneimine; Proteinuria; Rats; Rats, Inbred Strains; Ruthenium Red; Staining and Labeling