[Rapid determination of creatinine in urine by capillary electrophoresis in a multiple injection mode].

The level of urinary creatinine in clinic diagnosis is one of the useful index of renal function, and is often used as a reference of some biochemical substance determined in random urine. In this paper, a rapid multiple-injection capillary electrophoresis(CE) method for the determination of urinary creatinine is reported. A 36 cm x 50 microns coated capillary, a phosphoric acid buffer(0.1 mol/L, pH 2.5)and UV-detector at 200 nm wavelength were used in this method. In comparing with single injection for five samples, the analysis time of five successive injections can save 20 min. By use of pyridine as internal standard, the correlation between the concentrations of creatinine and the corresponding peak height ratios of creatinine and pyridine is good(r = 0.9996), and the assay precision is acceptable. We also compared the CE method with the bio-analyser(Jeffe' kinetic), and the results showed a satisfactory correlation (r = 0.9773, n = 12) and a shorter analysis time.

Analysis of urinary and circulating FDP subfragments and subunits: application of the western-blot technique.

We applied the Western-blot technique for qualifying fibrin/fibrinogen degradation products (FDP) subfragments and subunits. With this technique we examined urine or serum samples from patients with glomerulonephritis and disseminated intravascular coagulation (DIC), in order to observe how primary or secondary fibrinolysis functioned under these pathological conditions. We also tested the antigenic recognition of several FDP antibodies with this technique. The results obtained were as follows: 1) FDP in serum samples from patients with DIC consisted of both fibrinogenolytic and fibrinolytic products; 2) Fibrinolysis was predominant in the serum from one patient with Henoch-Schönlein purpura nephritis, and its level increased after fibrinolytic therapy with urokinase; 3) Fibrinogen and fibrin polymers were always the major components of urinary FDP, although fibrinolytic products such as subfragment D-D dimer were detected in patients with glomerular disease and increased in acute exacerbation; 4) Anti-FDP D-D dimer monoclonal antibody (DD 3B6), which is usually considered to react specifically with cross-linked fibrin derivatives, also seemed to have cross-reactivity with non-crosslinked fibrin derivatives.

An immunohistochemical study of experimental disseminated intravascular coagulation (DIC).

The experiment was focused on clarifying changes in fibrin or fibrinogen related materials (FRMs) in blood, urine, and renal tissues of rats with disseminated intravascular coagulation (DIC). DIC was induced by a continuous infusion of massive volume of physiologic saline (100 ml) immediately after endotoxin injection. FRM response was checked by biochemical and histochemical examinations at various intervals. In the blood of DIC rats, platelet and fibrinogen levels initially decreased, followed by an increasing plasma fibrin degradation products (FDP). Parallel with elevation of blood FDP the percentage of glomeruli with FRMs increased. Thereafter, FRMs were observed in renal tubuli and urine. Our observations indicated that FRMs in renal tubuli were derived from glomerular capillaries via Bowman's space. In conclusion, in DIC the immunoenzymehistochemical (IEH) procedure appeared necessary for an accurate pathological diagnosis, and the presence of FRMs in renal tubuli appeared to be an important finding even in absence of FRMs in glomeruli.

Fibrinopeptide A in urine from patients with venous thromboembolism, disseminated intravascular coagulation and rheumatoid arthritis--evidence for dephosphorylation and carboxyterminal degradation of the peptide by the kidney.

Urinary fibrinopeptide A immunoreactivity was determined by radioimmunoassay using two anti-fibrinopeptide A sera with a different specificity in patients with venous thromboembolism, disseminated intravascular coagulation and rheumatoid arthritis. Elevated levels were frequently observed with both sera, and intravenous administration of heparin in patients with a thromboembolic disorder resulted in a decline of urinary fibrinopeptide A (FPA) concentrations to normal or nearly normal values. For both sera significant correlations with plasma levels were found although one of the sera reacted significantly better with the material in urine samples from these patients than the other (p less than 0.0001, n = 73). Analysis of urinary fibrinopeptide A immunoreactivity by high performance liquid chromatography (HPLC) provided evidence that A peptide material present in this body fluid was heterogeneous. In view of the characteristics of the antisera used in this study, data suggest that urinary FPA immunoreactivity consists to a large extent of carboxyterminally degraded FPA. Excretion of circulating FPA immunoreactive material through the kidneys apparently involves dephosphorylation and carboxyterminal breakdown of the A peptide. Since both synthetic and native phosphorylated or unphosphorylated fibrinopeptide A appeared to be stable in urine in vitro, an active role of the kidney in degrading the A peptide is likely.

Studies on the purification and characterization of human urinary plasminogen and plasmin.

A new method is described for the preparation of highly purified human plasminogen and plasmin with specific activity of 32 CTA units per mg of protein. With this method, the purification of the urinary plasminogen + plasmin antigenic materials from patients with chronic glomerulonephritis, disseminated intravascular coagulation syndrome and severe toxemia of pregnancy was performed, and the resulting highly purified proenzyme and enzyme were analyzed by immunoelectrophoresis, separative agar electrophoresis, gel filtration and SDS-gel electrophoresis. Our findings indicated that urinary plasmin reflects more closely the extent of intraglomerular fibrinolysis, while urinary plasminogen reflects non-selective proteinuria in patients with chronic glomerulonephritis or severe toxemia of pregnancy.

Severe glomerulonephritis complicated by coagulopathy: treatment with anticoaguland and immunosuppresive drugs.

Serial determinations, using plasma fibrinogen gel chromatography as well as standard methodology, demonstrated that six children with severe glomerulonephritis, characterized on renal biopsy by glomerular necrosis and crescent formation, had persistent evidence of intravascular coagulation. Based on these observations, therapy with anticoagulants and azathicoagulants and azathioprine was instituted for one year; treatment with anticoagulants was continued for a second year. Anticoagulant therapy was initiated with heparin, followed by oral anticoagulation with phenindione and dipyridamole. In contrast to our earlier experience with similar patients, each of the present patients improved. Urinalyses returned to normal and glomerular filtration rates to near normal values in all patients at the end of the treatment period and have remained so for up to 3.9 years since treatment has been completed. Post-treatment biopsies showed remarkable improvement, with virtually no glomerulosclerosis even in patients who had had a high incidence of glomerular crescents before treatment. It is suggested that the therapeutic regimen favorably influenced the natural history of disease and that plasma fibrinogen chromatographic findings may be helpful in selecting patients likely to benefit from the use of anticoagulant therapy.

Acute intravascular hemolysis with minimal renal impairment in Clostridium perfringens infection.

We report here a case of post-abortal clostridium prefringens infection in which there was severe intravascular hemolysis with black urine, but only minor abnormalities of the clotting mechanism and mild renal failure. The patient recovered following supportive therapy only.