muramidase and Uremia

Plasma levels of granulocyte elastase in complex with alpha 1-proteinase inhibitor during hemodialysis were investigated in 15 patients (37.4 +/- 3.2 years) undergoing maintenance hemodialysis (47.0 +/- 12.9 months) with dialyzers made from cellulose hydrate, cuprophan, polymethylmethacrylate, ethylene-vinyl alcohol copolymer, and polyacrylonitrile. Cellulose hydrate membrane caused a maximal increase of the plasma levels of granulocyte elastase in complex with alpha 1-proteinase inhibitor (E-alpha 1PI: 1,659.0 +/- 256.8 ng/ml). Patients dialyzed with polyacrylonitrile dialyzers failed to exhibit comparable plasma levels of granulocyte elastase (E-alpha 1PI: 237.8 +/- 22.9 ng/ml). During hemodialysis plasma E-alpha 1PI values rose to a peak 643.0 +/- 174.7 ng/ml in patients on polymethylmethacrylate dialyzers, to 557.5 +/- 120.0 ng/ml on cuprophan dialyzers, but to only 381.9 +/- 54.0 ng/ml on ethylene-vinyl alcohol copolymer dialyzers. Plasma lysozyme levels decreased significantly in the presence of polyacrylonitrile and polymethylmethacrylate membranes. We conclude that the degree of PMNs stimulation depends on the nature of the dialyzer membrane material. The following membranes induce a reaction of increasing intensity: polyacrylonitrile, ethylene-vinyl alcohol copolymer, cuprophan, polymethylmethacrylate, and cellulose hydrate.

Topics: Acrylic Resins; Adult; Aged; alpha 1-Antitrypsin; Blood Proteins; Cellulose; Female; Granulocytes; Humans; Male; Membranes, Artificial; Middle Aged; Muramidase; Pancreatic Elastase; Polymethacrylic Acids; Polyvinyls; Renal Dialysis; Uremia

Diabetic uremic sera contain excessive amounts of reactive advanced glycation endproducts (AGEs), which accelerate the vasculopathy of diabetes and end-stage renal disease. To capture in vivo-derived toxic AGEs, high affinity AGE-binding protein lysozyme (LZ) was linked to a Sepharose 4B matrix. Initial studies showed that > 80% of 125I-AGE-BSA was retained by the LZ matrix, compared with < 10% retained by a control matrix. More than 60% of AGE-lysine was captured by the LZ matrix, and the LZ-bound fraction retained immunoreactivity and cross-linking activity, but had little intrinsic fluorescence (370/440 nm). After passage through the LZ matrix, AGE levels in diabetic sera (0.37+/-0.04 U/mg) were significantly reduced to a level (0.09+/-0.01 U/mg; n = 10; P < 0. 0001) comparable with the level of normal human serum, whereas total protein absorption was < 3%. The AGE-enriched serum fraction exhibited cross-linking activity, which was completely prevented by aminoguanidine. Among numerous LZ-bound proteins in diabetic uremic sera, three major proteins "susceptible" to AGE modification were identified: the immunoglobulin G light chain, apolipoprotein J (clusterin/SP-40,40), and the complement 3b beta chain. These findings indicate that the LZ-linked AGE affinity column may serve as an efficient method for the depletion of toxic AGEs from sera, including specific AGE-modified proteins that may be linked to altered immunity, lipoprotein metabolism, and accelerated vasculopathy in renal failure patients with or without diabetes.

Topics: Adult; Aged; Amino Acid Sequence; Blood Proteins; Diabetic Nephropathies; Glycation End Products, Advanced; Humans; Immunoblotting; Middle Aged; Molecular Sequence Data; Muramidase; Serum Albumin, Bovine; Uremia

3-Deoxyglucosone (3-DG) has been identified as an intermediate of the Maillard reaction in vitro. We measured serum 3-DG levels using gas chromatography/mass spectrometry and found a marked elevation in serum 3-DG levels in uremic patients compared with healthy subjects. The uremic patients with diabetes showed significantly higher serum concentrations of 3-DG than those without diabetes. 3-DG was demonstrated to be a potent protein-cross-linking agent in the reaction with lysozyme, leading to browning, fluorescence formation and polymerization of the protein by formation of advanced glycation end products (AGE). The increase in serum 3-DG levels in the uremic patients suggests that 3-DG may be responsible for the development of uremic complications by promoting the formation of AGE.

Topics: Adult; Aged; Blood Proteins; Cross-Linking Reagents; Deoxyglucose; Diabetic Nephropathies; Female; Gas Chromatography-Mass Spectrometry; Humans; In Vitro Techniques; Maillard Reaction; Male; Middle Aged; Muramidase; Uremia

By means of the immunocytochemical method, the level of cytoplasmic lysozyme in leukocytes from healthy volunteers (n = 50) and from patients with uremia (n = 50), leukocytosis (n = 50), various forms of leukemia (n = 36) and myelodysplastic syndrome (MDS) (n = 7) were analysed, and compared with that of simultaneously assayed serum lysozyme. Both the cytoplasmic and serum levels of lysozyme in uremia and leukocytosis were significantly higher than normal subjects (p < 0.001). No correlation, however, was found between their cytoplasmic and serum levels of lysozyme. Morphological analysis for various kinds of leukemia and MDS indicated that myelocytic and monocytic cells became highly positive for lysozyme staining with maturation, and that lymphocytes, leukemic myeloblasts and monoblasts were negative. The cytoplasmic and serum lysozyme levels of leukemias or MDS having a number of lysozyme-positive cells were elevated as compared with those of normal individuals. Among them acute myelocytic leukemia (FAB M4) revealed an excellent correlation between the lysozyme levels in cytoplasm and in serum. The rest whose serum lysozyme level tend to be lower than the cytoplasmic one gave poor correlation. Thus, serum lysozyme level is not fully reflected by the cytoplasmic level. The dual determination of cytoplasmic and serum lysozyme is suggested to be helpful on estimating leukemia types, the degree of cellular maturation and total cell mass, and might also provide a valuable tool for prediction of prognosis for these disorders.

Topics: Cytoplasm; Humans; Leukemia; Leukocytes; Leukocytosis; Muramidase; Myelodysplastic Syndromes; Uremia

In 10 chronic uraemics activities of some lysosomal enzymes were determined in peripheral blood neutrophils and plasma during haemoperfusion with charcoal cartridge Adsorba 300 C (Gambro). Blood plasma activities of arylsulfatases, beta-N-acetylglucosaminidase and beta-glucuronidase did not significantly change during haemoperfusion, while lysozyme activity was significantly increasing. Neutrophil contents of these enzymes decreased. The determinations of these enzyme activities revealed a positive outlet-inlet difference in blood plasma and a negative outlet-inlet difference in neutrophils during the first 20 minutes of the procedure. The results suggest that during haemoperfusion degranulation of the peripheral blood neutrophils occurs.

Topics: Acetylglucosaminidase; Adult; Arylsulfatases; Charcoal; Chronic Disease; Glucuronidase; Hemoperfusion; Humans; Muramidase; Neutrophils; Uremia

Topics: Acrylic Resins; Cellulose; Humans; In Vitro Techniques; Muramidase; Renal Dialysis; Uremia

Transient reduction in circulating polymorphonuclear granulocytes and eosinophils were observed early in hemodialysis. About a threefold increase in serum-lactoferrin occurred 2 h from the start of hemodialysis. Increments of the serum levels of eosinophil cationic protein (ECP) were observed as early as 1 h after initiation of hemodialysis, reaching maximum levels (about a fourfold increase from initial levels) 1 h later. When fresh blood was circulated through a dialyzer without having a patient in the circuit considerable increases of lactoferrin and ECP were also found. The intracellular contents of lactoferrin and ECP in granulocytes isolated from peripheral blood were unaffected throughout the dialysis period. Sera obtained at different times during dialysis induced no release of granular proteins from isolated granulocytes in vitro. The raised serum concentrations of lactoferrin and ECP during dialysis suggest that a local degranulation of neutrophils and eosinophils may take place probably in the dialyzer.

Topics: Adult; Blood Proteins; Eosinophil Granule Proteins; Eosinophils; Humans; Kidneys, Artificial; Lactoferrin; Lactoglobulins; Muramidase; Neutrophils; Renal Dialysis; Ribonucleases; Uremia

Topics: Animals; Female; Humans; Male; Middle Aged; Muramidase; Pelvic Neoplasms; Prostatic Hyperplasia; Rabbits; Uremia; Urinary Calculi

Topics: Acute Disease; Adult; Colitis, Ulcerative; Crohn Disease; Female; Humans; Leukemia, Monocytic, Acute; Male; Middle Aged; Muramidase; Uremia

Topics: Bacteriuria; Glomerular Filtration Rate; Glomerulonephritis; Humans; Hypertension; Kidney Diseases; Muramidase; Pyelonephritis; Uremia; Urinary Calculi

Topics: Animals; Chromatography, Ion Exchange; Humans; Kidney Transplantation; Kidney Tubular Necrosis, Acute; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Mice; Mice, Inbred BALB C; Molecular Weight; Muramidase; Proteinuria; Renal Dialysis; Transplantation, Homologous; Uremia

Topics: Albuminuria; Animals; Chromatography, Gel; Chromatography, Ion Exchange; Fibrin; Fibrinogen; Glomerular Filtration Rate; Glomerulonephritis; Humans; Immunoelectrophoresis; Kidney Transplantation; Molecular Weight; Muramidase; Nephrosis; Plasminogen; Rabbits; Streptokinase; Transplantation, Homologous; Uremia

On the assumption that increased urinary lysozyme concentration (;lysozymuria') indicates tubular proteinuria and therefore impaired tubular function, urinary lysozyme has been estimated in acute disorders where transient disturbances of renal function might be expected, in cases diagnosed clinically as extrarenal uraemia, and in a few examples of acute renal disease. Reversible lysozymuria occurred with hypokalaemia, postoperative ;collapse', electrolyte depletion, severe extrarenal infection, acute pyelonephritis, the nephrotic syndrome, after a few apparently uncomplicated surgical operations, and very transiently after ventricular fibrillation abolished by DC shock. There was no lysozymuria with severe uraemic heart failure, aspirin and paracetamol poisoning, or severe jaundice, nor in two cases of acute glomerulonephritis. Although lysozymuria may occasionally be useful in the clinical diagnosis of acutely disordered renal function, the results suggest that its value is limited; on the other hand, they have provided information on renal pathophysiology in acute disease.

Topics: Acetaminophen; Acute Disease; Aspirin; Electroshock; Glomerulonephritis; Heart Failure; Humans; Hypokalemia; Jaundice; Kidney; Kidney Diseases; Kidney Failure, Chronic; Kidney Tubules; Muramidase; Myocardial Infarction; Nephrotic Syndrome; Pneumonia; Postoperative Complications; Proteinuria; Pyelonephritis; Uremia; Ventricular Fibrillation

Topics: Adult; Bone Marrow; Butanols; Creatinine; Freezing; Humans; Leukocyte Count; Meningitis; Methods; Muramidase; Myeloproliferative Disorders; Neutrophils; Osteomyelitis; Pneumonia; Staphylococcal Infections; Ultrasonics; Uremia

Topics: Blood Coagulation; Factor XII; Humans; Muramidase; Uremia

Topics: Adult; Aged; Alpha-Globulins; Beta-Globulins; Blood Protein Electrophoresis; Female; Humans; Immune Sera; Kidney Diseases; Male; Metabolic Clearance Rate; Middle Aged; Muramidase; Uremia

Topics: Adult; Chronic Disease; Female; Humans; Kidney Failure, Chronic; Kidney Transplantation; Lymph; Male; Middle Aged; Muramidase; Renal Dialysis; Transplantation, Homologous; Uremia

The present study was directed toward determining the role of the kidney in the metabolism of various classes of serum proteins and to define the urinary protein excretion patterns and the pathogenesis of disorders of protein metabolism in patients with proteinuria. To this end, the metabolic fates of a small protein, lambda-L chain (mol wt 44,000), and a protein of intermediate size, IgG (mol wt 160,000), were studied in controls and patients with renal disease. Controls metabolized 0.28%/hr of circulating IgG and 22.3%/hr of circulating lambda-L chain. All the IgG and 99% of the lambda-L chain was catabolized with the remaining lambda-L chain lost intact into the urine. The kidney was shown to be the major site of catabolism for small serum proteins. Three distinct disorders of protein metabolism were noted in patients with renal tubular disease and tubular proteinuria, glomerular disease (the nephrotic syndrome), and disease involving the entire nephrons (uremia), respectively. Patients with renal tubular disease had a 50-fold increase in the daily urinary excretion of 15-40,000 molecular weight proteins such as lysozyme and lambda-L chains. Serum IgG and lambda-L chain survivals were normal; however, the fraction of the over-all lambda-L chain metabolism accounted for by proteinuria was increased 40-fold whereas endogenous catabolism was correspondingly decreased. Thus, tubular proteinuria results from a failure of proximal tubular uptake and catabolism of small proteins that are normally filtered through the glomerulus. Patients with the nephrotic syndrome had a slight increase in lambda-L chain survival whereas IgG survival was decreased and the fraction of IgG lost in the urine was markedly increased. Here, abnormal glomerular permeability to proteins of intermediate size is the basic abnormality. Patients with uremia had a normal IgG survival but a four to 10-fold prolongation of lambda-L chain survival due to loss of entire nephrons, the major site of metabolism of these proteins. This results in an increase (up to 10-fold) in the serum concentration of lambda-L chain, lysozyme, and other small biologically active proteins, a phenomenon that may be of importance in causing some of the manifestations of the uremic syndrome.

Topics: Adult; Blood Protein Disorders; Blood Protein Electrophoresis; Blood Proteins; Cell Membrane Permeability; Child; Electrophoresis, Disc; Humans; Immunoglobulin G; Iodine Radioisotopes; Kidney; Kidney Glomerulus; Molecular Weight; Muramidase; Nephrotic Syndrome; Proteinuria; Radioimmunoassay; Uremia

Serum levels, urinary excretion, and clearances of several proteins of different molecular weights were studied in 18 patients with mono- and myelomonocytic leukemia. Nine patients had normal renal function (group A) and nine had impaired renal function with azotemia (group B). The majority of patients in both groups had increased concentration of immunoglobulins, particularly IgG, IgA, and IgM; IgD level was normal. Serum transferrin and alpha(2)-macroglobulin were frequently reduced while the level of ceruloplasmin was often increased, especially in patients with azotemia. The activity of lysozyme in the serum was high in all patients, but was considerably higher in group B. Proteinuria was found in most patients but was more prominent in group B. Almost invariably albumin constituted less than 25% of the total protein excreted. Qualitative analysis of various urinary proteins by immunochemical techniques and clearance studies suggested the presence of glomerular as well as tubular dysfunction. Determination of urinary lysozyme frequently showed no direct correlation between the serum level of the enzyme and its concentration in the urine or its clearance by the kidney. In addition to glomerular filtration, impaired tubular reabsorption may account for the high level of lysozyme in the urine. It is postulated that the very high level of lysozyme in the glomerular filtrate and possibly hypergammaglobulinemia may play a role in the induction of tubular damage. Renal impairment has been correlated with histological changes in the kidneys. From a comparative study of various leukemias, it seems that the combined glomerular-tubular dysfunction is a manifestation unique to mono- and myelomonocytic leukemia.

Topics: Adult; Agammaglobulinemia; Aged; Albuminuria; Blood Chemical Analysis; Blood Proteins; Ceruloplasmin; Female; Humans; Hypergammaglobulinemia; Immunoglobulins; Kidney Glomerulus; Kidney Tubules; Leukemia, Myeloid; Male; Middle Aged; Muramidase; Nitrogen; Proteinuria; Transferrin; Uremia

Topics: Blood; Humans; Kidney Diseases; Muramidase; Proteinuria; Rats; Research; Urea; Uremia; Urine