alcian-blue and Nephrotic-Syndrome

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 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

It has been suggested that the negative charge on the surface of red blood cells (RBC) may reflect the charge on the glomerular capillaries. Measurements were made of RBC charge in patients with steroid-responsive nephrotic syndrome (SRNS) based on the binding of a cationic dye, alcian blue. There was no difference found between patients in relapse and in normal controls. We conclude that RBC charge measurements are not a reliable indicator of the glomerular charge barrier and a more specific method is required.

Topics: Adult; Alcian Blue; Basement Membrane; Child; Child, Preschool; Electrophysiology; Erythrocyte Membrane; Humans; Nephrotic Syndrome; Steroids

To elucidate the mechanism of enhanced ristocetin-induced platelet aggregation (RIPA) in steroid-responsive nephrotic syndrome (SRNS), plasma levels of von Willebrand factor antigen (vWF:Ag) and ristocetin cofactor (RCof) were examined in 6 patients and the amount of ristocetin-induced vWF binding to platelets was determined. At the initial or relapse stage, the plasma vWF:Ag level was 415 +/- 137% and the RCof level was 364 +/- 117%. The ratio of RCof/vWF:Ag was 0.90 +/- 0.15 and no abnormalities of vWF:Ag multimers were observed, indicating that neither functional nor structural abnormalities were present in patient's plasma. The amount of ristocetin-induced normal vWF binding to nephrotic washed platelets, when ristocetin was used at concentrations of 0.5, 0.75, and 1.0 mg/ml, was 152-163% above the binding to normal platelets. In addition, nephrotic washed platelets resuspended in either normal or nephrotic plasma aggregated at a low concentration of ristocetin (0.75 mg/ml) which did not induce aggregation of normal platelets. In accordance with these observations, the decrease of Alcian blue 8GX binding to platelets, reflecting diminished surface negative charge, was also observed. These results appear to indicate that the plasma vWF level and the altered surface-negative charge in platelets both contribute to heightened vWF binding to GPIb, thus lowering the ristocetin concentration required for RIPA in SRNS.

Topics: Adolescent; Adrenal Cortex Hormones; Alcian Blue; Antigens; Child; Child, Preschool; Female; Humans; Iodine Radioisotopes; Male; Nephrotic Syndrome; Platelet Aggregation; Platelet Membrane Glycoproteins; Protein Binding; Ristocetin; von Willebrand Factor

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

A simple chemical test, based on the binding of the cationic dye alcian-blue 8GX (AB), has been devised to measure negative charge on cell membranes. The test has demonstrated that AB binding to red blood cells and platelets is significantly less in children with steroid-responsive nephrotic syndrome (SRNS) or nephrotic syndrome associated with focal segmental glomerulosclerosis than in normal controls. However, the sialic acid content of the nephrotic cell membrane is normal. This suggests that a generalised loss of membrane negative charge occurs in SRNS and that this is due to neutralisation rather than absence of anionic groups.

Topics: Adolescent; Adult; Alcian Blue; Binding, Competitive; Blood Chemical Analysis; Cations; Cell Membrane; Child; Child, Preschool; Humans; N-Acetylneuraminic Acid; Nephrotic Syndrome; Sialic Acids