Urinary Fibrin/fibrinogen Degradation Products Research Articles

Measurement of urinary fibrin/fibrinogen degradation products by latex photometric immunoassay.

Intraglomerular coagulation and fibrinolysis could be involved in the exacerbation of renal diseases, and urinary fibrin/fibrinogen degradation products (FDP) may be applicable as an index. Although the urinary FDP can be estimated by the latex agglutination method, this technique has disadvantages such as a poor sensitivity and is of the semiassay type. Recently, a new method of measurement which improves on these disadvantages, termed the latex photometric immunoassay (LPIA) method, has been developed. However, since FDP measurement by LPIA was designed for the purpose of serum FDP estimation, a measurement technique for urinary FDP has not yet been established. The purpose of the present study was to devise a measurement procedure for urinary FDP employing the LPIA method, and to obtain data on the normal levels of urinary FDP in healthy subjects. The results obtained may be summarized as follows. (1) The urinary pH and coexistent substances such as bovine serum albumin, glucose, urea, bilirubin, ascorbic acid, and hemoglobin, did not influence the urinary FDP measurement. (2) No changes in urinary FDP were observed after 28-day storage at -20 degrees C or -80 degrees C in the presence or absence of tranexamic acid. (3) The coefficient of variation was 5.3%. (4) The normal level of FDP excretion was 3.33 +/- 7.95 micrograms/day. The present data demonstrated that the LPIA method enables the urinary FDP to be measured quantitatively with a good sensitivity.

膀胱癌の診断における尿中FDPの有用性に関する検討

We assessed the utility of urine fibrin/fibrinogen degradation products (FDP) as the screening test for bladder cancer. Single voided specimens were obtained from 87 consecutive patients (61 men and 26 women, mean age 70.7) on cystoscopy, and FDP, NMP22, BTA and cytology test were performed for the same specimens. Final diagnosis of bladder cancer was made by histological examination, which were compared with the results of above four screening methods. Histologically confirmed bladder cancer was found in 14 cases. Overall sensitivity of urinary FDP, NMP22, BTA and cytology were 79, 64, 36 and 36%, respectively. While the sensitivity of FDP was significantly higher than that of BTA and cytology, no significant difference was found between FDP and NMP22. Overall specificity of these four methods were 69, 78, 92 and 90%, respectively. The specificity of FDP and NMP22 were significantly lower than that of BTA and cytology, but satisfactory as a screening test. The sensitivity of the four methods for low-grade and non-invasive tumors were 70, 50, 30 and 10% (G1 or G2, n = 10), and 75, 58, 33 and 25% (Ta or T1, n = 12), respectively. FDP might have a high sensitivity for even low-grade and non-invasive tumors. FDP in voided urine is a good screening method for bladder cancer because of its high sensitivity for low-grade and non-invasive tumors, and its diagnostic ability could be superior to NMP22.

Significance of Urinary Fibrin/Fibrinogen Degradation Products in Renal Diseases Measured by a Highly Sensitive ELISA

The urinary fibrin/fibrinogen degradation products (FDP), as sensitive indicators of various renal disorders, have been measured by several methods. For their determination, a new and highly sensitive enzyme-linked immunosorbent assay not requiring the urine concentration has been developed. The study comprised 42 patients with nonnephrotic chronic glomerulonephritis (CGN), 23 patients with primary nephrotic syndrome (NS), and 29 healthy adults. The results were as follows: (1) the content of urinary FDP in normal subjects was 10.30 +/- 9.08 ng/ml; (2) the mean level of urinary FDP in both CGN and NS groups was significantly higher than in normal subjects; (3) in the CGN group itself there was a tendency for an increase of urinary FDP during more active forms of the disease, and (4) there was a significant correlation between urinary FDP and urinary protein in the CGN group, whereas no correlation was observed in the NS group. These results suggest that the major part of urinary FDP in the CGN group derives from the increased filtration, while its origin in the NS group is not related to increased filtration only, but may also have involved intraglomerular coagulation abnormalities. The newly developed enzyme-linked immunosorbent assay can detect urinary FDP levels lower than 3.9 ng/ml. Therefore, this method can be of great value in determining the degree of abnormalities of intraglomerular coagulation and fibrinolysis in renal diseases.

各種腎疾患における尿中cross-linked FDPについての検討

Fibrin/Fibrinogen degradation products (FDP) and cross-linked FDP (XLFDP) have been found in the urine of many patients with renal disease. FDP result from fibrinogenolysis and fibrinolysis. It is useful to detect the urinary XLFDP which results from fibrinolysis in order to diagnose intraglomerular coagulation. I investigated urinary FDP and XLFDP in patients with various renal diseases. Urinary FDP and XLFDP were detected by latex aggregation test in the urine of 96 patients. Urinary XLFDP was measured by enzyme-linked immunosorbent assay (ELISA) using monoclonal antibody in the urine of 140 patients. The positive rates of urinary XLFDP were higher in chronic renal failure, membranous nephropathy and membranous-proliferative glomerulonephritis than in other renal diseases. High levels of urinary XLFDP were found in membranous nephropathy, membranous proliferative glomerulonephritis and non-IgA nephropathy. There was an obvious relationship between urinary XLFDP and the degree of proteinuria, hematuria, serum creatinine and intraglomerular fibrin deposits. The high levels of urinary XLFDP were detected in the case of severe proteinuria, normal and mild hematuria, normal and slightly increased serum creatinine and mild intraglomerular fibrin deposits.

Significance of urinary fibrin/fibrinogen degradation products (FDP) measured by highly sensitive ELISA method in various renal diseases

In order to clarify the abnormalities of intra-glomerular coagulation and fibrinolysis in patients with various renal diseases, urinary fibrin/fibrinogen degradation products (FDP) have been examined by several methods. We established a highly sensitive new method of enzyme-linked immunosorbent assay for urinary FDP. The results were as follow: 1) The mean +/- SD of urinary FDP in normal subjects was 10.30 +/- 9.08ng/ml. 2) The urinary FDP levels in chronic glomerulonephritis, nephrotic syndrome and chronic renal failure patients were significantly higher than normal subjects, and the levels in SLE, Alport's syndrome patients were higher than normal subjects. 3) The urinary FDP levels were a little bit higher in the patients with proliferative glomerulonephritis than in chronic glomerulonephritis patients with minor lesion or membranous nephropathy. 4) There was significant correlation between urinary FDP and urinary protein in chronic glomerulonephritis, while there was no correlation in nephrotic syndrome. 5) There was no correlation between urinary FDP and intra-glomerular fibrin deposits examined by immunofluorescent study in chronic glomerulonephritis, while in nephrotic syndrome, there were high levels of urinary FDP in the positive fibrin deposits cases. These results suggested that the most of the part of excretion of the urinary FDP in chronic glomerulonephritis is associated with the filtration of blood FDP to urine through the glomerular basement membrane, while in the nephrotic syndrome cases the origin of urinary FDP is related to the filtration and/or the intra-glomerular coagulation abnormalities.

Analysis of Urinary and Circulating EDP 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.

Fragments of urinary fibrin/fibrinogen degradation products and cross-linked fibrin degradation products in various renal diseases

In children with several kinds of glomerular disease, fragments of fibrin/fibrinogen degradation products(FDP) and cross-linked fibrin degradation products(XLFDP) in the urine were investigated by autoradiography using western blotting method. Results were compared with selectivity of proteins observed in cases of proteinuria, or with histological findings. Patients with nephrotic syndrome exhibited slightly increased amount of urinary FDP, consisted mainly of X and Y fragments. On the other hand, in cases of proliferative glomerulonephritis, such as acute glomerulonephritis, purpura nephritis, Ig A nephropathy, systemic lupus erythematosus, or hemolytic uremic syndrome, increased FDP, including XLFDP, was detected in the urine. In these cases, FDP was consisted mainly of fragments X, Y, and D-dimer, and could not be correlated with the degree of mesangial proliferation or with urinary protein selectivity. It was concluded that the increased urinary FDP and XLFDP were derived not only from filtration of plasma fibrinogen or FDP, but also from fibrinolysis of intraglomerular fibrin deposits.

Urinary fibrin/fibrinogen degradation products in transitional cell carcinoma of the bladder.

Urinary fibrin/fibrinogen degradation products (FDPs) were measured in 210 specimens from 174 patients with newly or previously diagnosed transitional cell carcinoma of the bladder. They were detected in 94% of patients with deeply invasive bladder tumours (pT2-4) compared with 17% of superficial tumours. Microalbuminuria (greater than 50 micrograms/g creatinine) was also found in 80% of patients with pT2-4 lesions. Both were compared with urine cytology. Urinary FDPs are markers of bladder tumour invasion. Our results suggest that urinary FDPs are not of value in screening for the presence of bladder neoplasia but their role may be in following patients with superficial bladder tumours to detect those tumours which become invasive. The mode of excretion of the FDPs in the urine is discussed.

小児じん疾患におけるじん組織フィブリンとＦＤＰフラグメントについて

In order to elucidate the relation between intraglomerular fibrin deposition and types of serum and urinary fibrin/fibrinogen degradation products (FDP) in children with various renal diseases, the extent and distribution of fibrinogen related antigen (FRA) deposition in the kidney tissue which was perfesed with monochloacetic acid (MCA) solution was examind by immunofluorescent microscopy in comparison with deposition of factor XIII subunit-A (XIII-a). In addition to the immunofluorescent studies, the typing of FDP was made by SDS polyacryamide gel electrophoresis and crossed immunoelectrophoresis (SDS-PAGE-CIE).Intrarenal deposition of FRA after MCA treatment was observed mainly within the glomerular capillary walls, mesangium or vessels, and its extent and distribution showed a good agreement with that of XIII-a. Patterns of SDS-PAGE-CIE of MCA insoluble FRA products by plasmin were similar to those seen in cross-linked fibrin. These findings suggest that MCA insoluble FRA or XIII-a deposition reflects the extent and distribution of cross-linked fibrin.The presence of D-dimer in the urinary FDP was well seen in the children who had intrarenal fibrin deposition.

Fibrin deposits in the glomeruli and fibrin/fibrinogen degradation products (FDP) in the blood and urine in some renal diseases.

In 115 patients with various glomerular diseases and in 23 with chronic pyelonephritis the comparative incidence of glomerular fibrin deposits, and blood and urinary FDP was studied for evaluation of their clinical value. The results of the study indicate that the determination of urinary FDP is the most reliable clinico-laboratory test for the presence of increased intrarenal haemocoagulation. Moreover, the quantitative assessment of urinary FDP could be also used as an index for estimating the activity of the pathological process, the selectivity of proteinuria, the need for anticoagulant therapy, and the prognosis of the disease.

Urinary fibrin-fibrinogen degradation products and intraglomerular fibrin-fibrinogen deposition in various renal diseases

The typing of the urinary fibrin-fibrinogen degradation products (FDP) and the deposition of fibrin-fibrinogen, factor XIII-subunit A (XIII-A) and factor XIII-subunit S (XIII-S) in the glomeruli were investigated in the children with several kinds of renal diseases. In acute glomerulonephritis with diffuse proliferative lesion, the urinary FDP were composed of fragments X,Y,D and E. There was no D-dimer fragment detected in the urine, although, mild deposition of fibrin-fibrinogen, XIII-A and XIII-S in the glomeruli was observed. The urinary FDP may be derived from plasma fibrinogen or serum FDP. In nephrotic syndrome with diffuse proliferative or focal global sclerosing lesion and in purpura nephritis with mesangial proliferation with crescents, the urinary FDP consisted of fragments X and Y, and lacked in D-dimer. In these cases, the major part of fibrin-fibrinogen in the glomeruli might be fibrinogen or non-cross-linked fibrin, and FDP excretion might arise from glomerular permeability or from lysis of fibrinogen or non-cross-linked fibrin. In one patient with hemolytic uremic syndrome and in another patient with systemic lupus erythematodes nephropathy, intense deposition of fibrin-fibrinogen, XIII-A and XIII-S within the glomerular capillaries or mesangial area were observed. In the urine of those cases, D-dimer was detected, which seems to reflect the lysis of cross-linked fibrin in the glomeruli.

Origin of Urinary Fibrin-Fibrinogen Degradation Products in Renal Glomerular Disease

The origin and mechanism of renal clearance of urinary 'fibrin-fibrinogen degradation products' (FDP) were studied in patients with renal glomerular diseases associated with heavy, non-selective proteinuria and high levels of urinary FDP. The results indicated that the urinary FDP arose primarily by the filtration of unaltered plasma fibrinogen through a damaged and abnormally permeable glomerular basement membrane and that a variable degree of lysis of the filtered fibrinogen occurred in the urine. The lysis of cross-linked fibrin in intraglomerular deposits, as evidenced by the presence of dimeric fragment D in the urine, appeared to contribute only a small amount to the total urinary FDP excretion.

The relationship between urinary fibrin/fibrinogen degradation products (U-FDP), and the effects of warfarin and heparin administration to these parameters in renal disease

The relationship between urinary fibrin/fibrinogen degradation products (U-FDP), and the effects of warfarin and heparin administration to these parameters in renal disease

Fibrin/fibrinogen degradation products (FDP) in urine and serum after prolonged heavy exercise.

The presence of FDP in the urine, known to be a sensitive indicator of various kidney disorders, was studied in 14 well-trained men participating in a 70-km cross-country ski race. None of the urine samples contained FDP on the day before the race. Immediately after the race, 4 samples contained FDP. High molecular weight FDP were found in 2 of them, whereas 2 others contained low molecular weight products only. 2 days after the race, 1 of these subjects still had FDP in the urine. In addition, 2 newly FDP positives were observed. Serum FDP were slightly elevated in 2 subjects before the race, and in another subject immediately after the race. The presence of urinary FDP did not correlate either with urinary albumin or uromucoid, or with serum FDP. A drop in plasma fibrinogen immediately after the race was noted in all subjects. It is suggested that the present observations may reflect a transient hyperproteolysis (coagulation and fibrinolysis) in the glomerular circulation, including fibrin formation and dissolution, associated with a transient damage of glomerular capillaries.

Determination of urinary fibrin/fibrinogen degradation products by radioimmunoassay.

Glomerular fibrin/fibrinogen related material is often found in the early stage of especially proliferative types of glomerulonephritis and during rejection of renal grafts after transplantation. The presence and concentration of urinary fibrin/fibrinogen degradation products (FDP) have been found to be correlated to the extent of glomerular fibrin deposits. Most methods for FDP determination hitherto available are not sensitive enough to detect such small amounts of the fibrin/fibrinogen related material in the urine as are found in early glomerulonephritis. A radioimmunoassay for determining urinary FDP is described, in which the high molecular weight degradation products (HMWDP) are first converted into end degradation products (D and E) by addition of plasminogen and streptokinase to the urine sample. In the test an antiserum against purified D-products is used, which avoids the influence of varying binding properties and therefore makes the method more specific.

Urinary fibrin-fibrinogen degradation products in nephrotic syndrome.

The urinary concentration of fibrin-fibrinogen degradation products (F.D.P.) was measured in 90 patients with proteinuria above 2 g/1 and correlated with proteinuria, differential protein clearances, serum urea and creatinine, and renal biopsy findings. There was a linear correlation (r equals 0-7; P less than 0-001) between the urinary F.D.P. excretion and the selectivity of the proteinuria such that patients with highly selective proteinuria excreted only small amounts of F.D.P. whereas those with non-selective proteinuria excreted much higher levels. There was a significant correlation between the urinary F.D.P. excretion and the urine:serum (U:S) ratio of IgG excretion but not with the U:S ratio or urinary excretion of albumin or transferrin. Sephadex G200 column chromatography of the concentrated urine in 26 cases showed that patients with highly selective proteinuria excreted predominantly F.D.P. of low molecular weight in the urine whereas those with non-selective proteinuria excreted mainly fibrinogen and products of high molecular weight. Hence the type and quantity of F.D.P. in the urine are determined primarily by the differential filtration of fibrinogen and the various degradation products from the plasma through the glomerular basement membrane, which in turn is determined by the "pore size" of the basement membrane. In clinical nephrology measurement of the urinary F.D.P. level provides a rapid and convenient means of estimating the differential protein clearance.

Urinary tract fibrinolysis

Tissue samples from all portions of the urinary collecting system were assayed for the presence of tissue fixed plasminogen activator activity. Daily urine samples were obtained from 17 patients following urinary tract surgery and assayed for the presence of fibrin-fibrinogen degradation products (FDP). Hematuria was present in all samples. Urine samples were obtained from 17 patients with chronic renal disease and proteinuria and similarly assayed for FDP. All portions of the urinary collecting system contained considerable tissue fixed plasminogen activator activity. All patients with hematuria had FDP present in the urine, indicating that fibrinolysis is the process by which the clots are removed from the urinary tract. These results indicate that correlation of urinary FDP with renal parenchymal disease should be attempted only in the documented absence of hematuria.

Fibrin-fibrinogen degradation products in children with renal disease.

Fibrin/fibrinogen degradation products (FDP) were measured in the serum and urine of children with various forms of renal disease. Serum FDP was raised both with nephrosis and with active proliferative nephritis. Urine FDP was rarely present in nephrosis but was significantly increased during the active phase of proliferative nephritis and also in urinary tract infection with frank haematuria. Urinary FDP correlated with total urinary protein in proliferative nephritis but not in nephrosis, nor did it correlate with serum FDP in either condition. The major application of urinary FDP determination in clinical practice is as an indicator of activity and possible response to treatment in the management of active proliferative nephritis.

Fibrin-fibrinogin degradation products in cardiovascular surgery.

Two hundred fifty-five determinations of the protamine sulfate test, the staphylococcal clumping test, and the tanned red blood cell hemagglutination inhibition immunoassay were performed on 30 patients undergoing cardiovascular surgery. The level of serum fibrin-fibrinogen degradation products rose in all patients and reached the highest levels between the fifth and ninth postoperative day. After the first postoperative day, a fibrin-fibrinogen degradation product increase of greater than 10μg/ml within 24 to 48 hours was associated with clinically recognized thrombotic complications in only two out of five patients. Isolated levels of serum fibrin-fibrinogen degradation products as high as 67μg/ml could not be correlated with clinically evident thrombotic disease or serious complications of surgery. Increased excretion of urinary fibrin-fibrinogen degradation products was associated with, but not predictive of, acute ischemic renal failure.

Urinary fibrin-fibrinogen degradation products (FDP) in renal diseases and during thrombolytic therapy.

Using gel filtration on Sephadex G-200 and the immunochemical method of Bouma et al., the urinary FDP in uraemia and after renal transplantation were found to be mainly of high molecular weight type. Their concentration did not vary with the urinary concentration of lysozyme or albumin. The findings suggest that the urinary FDP are not the results of glomerular filtration or further degradation in the lower urinary tract of filtered fibrinogen. The urine from patients receiving thrombolytic therapy contained low molecular weight products. They were thought to result from an overloading of the tubular reabsorption mechanism.