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Background: The aim of the present study was to examine the presence of motilin in the duodenal epithelial cells of premature infants of

Background: The proteinuria resulting from IgA nephropathy is secondary to altered charge-selective and/or size-selective properties of the glomerular capillary walls. However, the functional changes occurring within the glomerular capillary network which lead to proteinuria are still poorly understood. In this study, we analyzed the participation of charged components in the glomerular capillary and their role with respect to proteinuria in childhood IgA nephropathy. Methods: We examined glomerular anionic charge in renal biopsy specimens with confocal laser scanning microscopy using FITC-conjugated poly-L-lysine as a cationic tracer. The renal specimens investigated were from 9 children with IgA nephropathy (IgAN(+)) associated with detectable proteinuria in a morning urine specimen, 8 children with IgA nephropathy (IgAN(–)) and 11 children with thin basement membrane disease (TBMD) with no detectable proteinuria. Results: The labeling intensity of glomerular fixed anionic sites from IgAN(+) was significantly lower than that of IgAN(–) and TBMD. Staining the serial sections following methylation treatment revealed that the labeling intensity for fixed anionic sites in TBMD was significantly higher than that of both IgAN(+) and IgAN(–). On the other hand, saponification treatment resulted in significantly more intensive fluorescence in TBMD and IgAN(–) than in IgAN(+). Furthermore, statistical analysis demonstrated a significant correlation between 24-hour protein excretion and the intensity of fixed anionic sites in all patients with IgA nephropathy at pH 7.0 and following staining with saponification treatment. Conclusions: These findings suggest that a reduction of glomerular anionic charge might be causally associated with the development of proteinuria in childhood IgA nephropathy. Furthermore, sulfate components such as heparan sulfate proteoglycan might be involved initially followed by carboxyl components such as sialoglycoproteins in the glomeruli of patients with IgA nephropathy contributing to the occurrence of proteinuria. We suggest that this method represents a straightforward approach to dissect the participation of charged components in the pathogenesis of childhood IgA nephropathy and their relationship to the development of glomerular proteinuria.

Precise localization of cytokines such as transforming growth factor-beta (TGF-beta), tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1 and IL-6 was observed in glomeruli using immunogold electron microscopy in 21 children with various types of renal diseases. The distribution pattern of these cytokines, as well as immunoglobulins, C3c and fibrinogen (Fg), was essentially confined to the electron-dense deposits (EDDs) regardless of their location. Frequency of positive labelling of each cytokine was different among various types of renal disorder, that is, TGF-beta was found mainly in lupus nephritis (LN), membranous nephropathy and IgA nephropathy, TNF-alpha in LN, and IL-1 in Henoch-Schönlein purpura nephritis. IL-6 was detected only in 1 case of LN. TNF-alpha was also found in the cytoplasm of glomerular epithelial cells. Furthermore, in order to evaluate the relation of cytokines to mesangial expansion, extracellular matrix components such as type IV collagen, laminin and fibronectin were stained. The result was that there was no significant correlation between the signal intensity or distribution pattern of cytokines and that of extracellular matrix components. These findings indicate that these cytokines could be associated with the formation of EDDs together with immunoglobulins, C3c and Fg. The involvement of each cytokine in renal pathophysiology might also depend upon the type of renal disease. They also raise the possibility that the glomerular epithelial cells might produce or absorb TNF-alpha. However, these results did not show significant correlation between cytokine involvement and mesangial expansion.

A report is presented of a patient with neonatal erythema infectiosum who developed petechiae, transient thrombocytopenia and transient cardiac failure due to transplacental transmission of human parvovirus B19 (HPV B19) infection. It is suggested that the thrombocytopenia was caused by platelet-associated IgG produced by the patient, and that the cardiac failure may have been caused by direct entry of HPV B19 into the cardiac tissue.

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 ± 1.17%. 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.

Type III Bartter syndrome (BS) (OMIM607364) is caused by mutations in the basolateral chloride channel ClC-Kb gene (CLCNKB). The CLCNKB gene is sometimes reported as having a large deletion mutation, but all cases reported previously were large homozygous deletions and a large heterozygous deletion is impossible to detect by direct sequencing. This report concerns a genetic analysis of five Japanese patients with type III BS. To identify the mutations, we used polymerase chain reaction (PCR) and direct sequencing. To detect large heterozygous deletion mutations of the CLCNKB gene, we conducted semiquantitative PCR amplification using capillary electrophoresis. The result was that four mutations were identified, comprising one novel 2-bp deletion mutation, an entire heterozygous deletion, and a heterozygous deletion mutation of exons 1 and 2. The nonsense mutation W610X was detected in all patients, and this mutation is likely to constitute a founder effect in Japan. Capillary electrophoresis is a new method and extremely useful for detecting large heterozygous deletions, and should be used to examine type III BS cases in whom only a heterozygous mutation has been detected by direct sequencing. This is the first report to identify large heterozygous deletion mutations in the CLCNKB gene in patients with type III BS.

Aim: Although infiltrating macrophages found in renal biopsy specimens have been accepted as a useful marker for evaluating the activity of IgA nephropathy (IgAN), it is difficult to perform renal biopsies repeatedly, especially in children. To establish a more convenient and noninvasive method for estimating the degree of macrophage infiltration we examined the number of macrophages in urinary sediments. Patients and methods: Ten ml of morning urine were collected from 30 children with IgAN, 10 with thin basement membrane disease (TBMD), 8 with idiopathic renal hemorrhage (IRH) which was defined as nonglomerular hematuria due to nutcracker phenomenon revealed on ultrasonography, and 10 healthy children as controls. Ten of the 30 children with IgAN were treated with combination therapy comprising prednisolone, warfarin and dipyridamole and urine samples were collected weekly during the period oftreatment. Two μl of the urine sediment were smeared on glass slides, dried and stained with a monoclonal antibody to human macrophages (anti-CD68, PG-M 1) followed by a FITC-conjugated secondary antibody. After staining with propidium iodide (PI), the cells were examined by fluorescence microscopy with cells stained with both FITC and PI being counted as macrophages. In addition, anti-CD68 staining was used to quantify macrophage infiltration in renal biopsies from the same group of IgAN patients. Results: The number of urine macrophages in children with IgAN was significantly higher than in children with TBMD and I RH as well as the control group (p < 0.01), whereas that was similar among TBMD, IRH and healthy children. In IgAN, there was a significant correlation between urine macrophage number and the activity index (p < 0.01), proteinuria (p < 0.01) and urine WBC count (p < 0.01). In addition, there was also a significant correlation between urine macrophage number and glomerular (p < 0.05) as well as interstitial macrophage infiltration (p < 0.01). In children with IgAN who received combination therapy, urine macrophage number decreased significantly (p < 0.01) in the 1st week of treatment whilst the degree of proteinuria decreased significantly (p < 0.01) in the 4th week. Conclusion: Urinary macrophage number may represent a noninvasive and straightforward estimate of the pathological activity evident in renal biopsy specimens, and may also be a more sensitive indicator than proteinuria of the therapeutic effect of interventional treatments in childhood IgAN.

The current hypothesis for the pathogenesis of childhood idiopathic nephrotic syndrome (INS) favors the involvement of a T cell-mediated immune response. Various cytokines derived from T cells may play a critical role in INS. Previous studies have measured serum or urine cytokine levels and suggest an imbalance of the T cell-mediated immune response. To elucidate the true profile of T cell-derived cytokines, we determined interleukin (IL)-2, interferon (IFN)-gamma, IL-4, IL-10, and tumor necrosis factor (TNF)-alpha mRNA expression in children with INS. We collected mRNA from peripheral blood mononuclear cells together with plasma and urine from nine children in the acute and remission phases of INS. Expression of IL-2, IFN-gamma, IL-4, IL-10, and TNF-alpha mRNA was determined by a quantitative real-time PCR assay. Plasma and urine cytokine concentrations were measured using a specific enzyme-linked immunosorbent assay. These data were compared between the acute and remission phase in the same patients. The IL-2 mRNA levels were significantly higher in the acute phase than in the remission phase, whilst no significant difference was found in the other cytokines investigated. There was no significant difference in the plasma and urine cytokine concentrations between the acute and remission phase. Our results indicate increased expression of IL-2 mRNA in the acute phase of INS, suggesting that IL-2, at least in part, might be involved in the pathophysiology of childhood INS.

Thin basement membrane disease (TBMD) and Alport syndrome, two forms of childhood nephritis, have generally been considered to be hereditary diseases. In Alport syndrome, several reports have demonstrated pathogenic mutations of the genes encoding type IV collagen alpha3, 4 and/or 5 chain [alpha3, 4 and/or 5(IV)]. Previous immunohistochemical studies indicated that these antigens were absent from the glomerular basement membrane (GBM) in Alport syndrome, whilst a normal labeling pattern was maintained in TBMD. In order to understand the role of the alpha3, 4 and/or 5(IV) antigens in TBMD, we used confocal laser scanning microscopy (CLSM) to examine cryosections of renal biopsies from 12 children with TBMD and 11 control children with IgA nephropathy (IgAN) without proteinuria. All tissue sections were stained with a mixture of FITC-conjugated rat monoclonal antibodies directed against human alpha3(IV), alpha4(IV) or alpha5(IV) and a Texas red-conjugated rat monoclonal antibody raised against human alpha2(IV). CLSM was performed and quantitative analysis of the ratio of the staining signal for alpha3(IV), alpha4(IV) or alpha5(IV) to alpha2(IV) [alpha3(IV), alpha4(IV) or alpha5(IV)/alpha2(IV)] along the GBM was determined. The average number of pixels for alpha3(IV), alpha4(IV) or alpha5(IV)/alpha2(IV) was 3.52 +/- 1.49, 3.54 +/- 1.25 and 1.09 +/- 0.49 in TBMD and 3.62 +/- 1.46, 3.99 +/- 1.53 and 1.77 +/- 0.47 in control subjects, respectively. Statistical analysis indicated that alpha5(IV)/alpha2(IV) ratio was significantly lower (p0.01) in children with TBMD compared to controls. These findings raise the possibility that TBMD might be caused by an abnormality of the alpha5(IV) antigen along the GBM.

Analysis of the hemostasis system using biochemical techniques in children with minimal change nephrotic syndrome (MCNS) has previously been restricted to in vitro assays. The recent introduction of measurement of shear stress-induced platelet aggregation (SIPA) using platelet-rich plasma (PRP) has facilitated detailed investigation of the hemostatic system in vivo. In order to further analyze the etiology of the thrombotic tendency exhibited by patients with childhood MCNS, we investigated SIPA at both low shear stress (L-SIPA) and high shear stress (H-SIPA) in 14 children with MCNS using PRP collected weekly after commencing prednisolone therapy. Seven patients were newly diagnosed cases of MCNS (ND) whereas the remainder had suffered a disease relapse (DR). Prior to prednisolone therapy L-SIPA, which was thought to be affected by fibrinogen (Fbg) levels, was significantly increased in both patient groups compared to normal controls (17.4 +/- 4.1% vs. 3.6 +/- 0.7%, ND vs control, p0.01: 11.7 +/- 3% vs. 2 +/- 0.7%, DR vs control, p0.01) with values declining at weekly intervals thereafter. Plasma Fbg levels in simultaneously collected samples followed a similar course. In contrast, H-SIPA, which was thought to be affected by von Willebrand factor (VWF), was significantly enhanced in MCNS patients after 1 week of prednisolone therapy compared to the controls (45 +/- 5.1% vs. 26.3 +/- 3.5%, ND vs normal, p0.05: 36.9 +/- 3.3% vs. 25.5 +/- 1.6%, DR vs. normal, p0.05). Plasma ristocetin cofactor and VWF antigen levels in simultaneously collected samples followed a similar course, whereas thrombin-antithrombin complex (TAT) levels remained constant. These results indicate that SIPA is enhanced in the acute stage of childhood MCNS, especially L-SIPA prior to the initiation of prednisolone therapy and H-SIPA after 1 week of prednisolone therapy, and that these phenomena may be affected by plasma Fbg and VWF levels, respectively. The enhanced SIPA may play an important thrombogenic role in the acute phase of childhood MCNS.

Background: This study was undertaken to determine the efficacy of low-dose intravenous erythromycin (EM) administration in infants with feeding intolerance. Methods: The subjects were 26 infants who would not accept enteral feeding within 5 days after birth. Fourteen infants (gestational age: 30.6 ′ 5.4 weeks and birthweight: 1466 ′ 825 g) were given EM intravenously at a dose of 1 mg/kg, three times daily (EM group). Doses were increased to 2 mg/kg in five infants who showed a poor response. Twelve infants (gestational age: 30.5 ′ 5.0 weeks and birthweight: 1317 ′ 672 g) were observed without EM administration (non-EM group). Blood concentrations of EM at 2 h after administration were measured on 8 (′ 2) days after the start of EM administration in the EM group. Results: Digestive perturbations and intestinal gasless and/or atonic shadows on X-ray findings markedly improved in the EM group soon after the treatment. Comparing the EM group and non-EM group, the postnatal ages at the start of successful enteral feeding were 9.1 ′ 3.2 days and 14.0 ′ 4.1 days, respectively (P < 0.01). The postnatal ages at feeding of 100 mL/kg per day were 15.2 ′ 4.0 days and 23.4 ′ 6.2 days, respectively (P < 0.01). The blood EM concentrations of 1 mg/kg and 2 mg/kg were 0.29 ′ 0.28 μg/mL and 0.57 ′ 0.20 μg/mL, respectively (P < 0.05). No adverse effect on cardiac status or in blood examinations was observed in any infant in the EM group. Conclusion: These results suggest that intravenous low-dose EM administration is a useful and safe treatment of feeding intolerance in infants including extremely low-birthweight infants.

BACKGROUND Activated neutrophils play an important role in the pathogenesis of renal injury in humans and in experimental models of hemolytic uremic syndrome (HUS). To evaluate the clinical significance of the circulating granulocyte elastase-alpha1-proteinase inhibitor complex (GEPIC), which is a marker of neutrophil activation, we investigated the plasma concentrations of GEPIC in children with hemolytic uremic syndrome (HUS) associated with verotoxin-producing Escherichia coli (VTEC), VTEC gastroenteritis without HUS and in normal controls. METHODS Of 22 children (1-19 years of age; mean age 5.5 years) with VTEC infection, nine were diagnosed with HUS. Plasma GEPIC, soluble thrombomodulin (sTM) and thrombin-antithrombin-III complex (TAT) levels were measured by ELISA. RESULTS The number of polymorphonuclear leukocytes and the levels of plasma GEPIC in patients with HUS were significantly higher than those in non-HUS (9850+/-5091 vs. 5278+/-3327 /microL, P

Between 1996 and 1997, 2,045 percutaneous renal biopsies were performed on native kidneys in 2,013 patients in pediatric nephrology units in Japan. Of these, 50.8% were performed by automated needle biopsy gun under ultrasound guidance, and the standard biopsy needle, Tru-cut needle or Vim-Silverman needle, under fluoroscopic guidance was used in 12.4% and 12.3% of the biopsies, respectively. Adequate renal tissue for histological diagnosis was obtained in 98.7% of cases, and the success rates for the techniques were not significantly different. The overall complication rate was 5.8%; gross hematuria occurred in 2.7% and large perirenal hematoma in 0.9% of cases. These complication rates were higher when a standard needle under fluoroscopic guidance was used compared with an automated needle under ultrasound guidance. We conclude that pediatric nephrologists in Japan perform percutaneous renal biopsies safely, partly due to technical improvements, such as the automated needle or ultrasound guidance.

The relationship between the activation of platelets (PLT) in urine and renal histological findings in children with IgA nephropathy (IgAN), thin basement membrane disease (TBMD), and minimal-change nephrotic syndrome (MCNS) was examined. The ratio of activated PLT to total PLT (activated and nonactivated PLT) was examined by double immunofluorescence using rhodamine-conjugated P selectin antibody (activated PLT) and fluorescein isothiocyanate conjugated PLT membrane glycoprotein antibody (GPIIb/IIIa, total PLT); the effect of urine on activation on PLT was also investigated. The number of activated PLT and the ratio of activated PLT to total PLT in urinary sediments were significantly higher in children with IgAN with diffuse mesangial proliferation than in those with TBMD or MCNS. PLT were activated by addition of urine in 13 out of 27 children with IgAN, and the activity was higher in the urine of those with active glomerular or interstitial lesions, while the urine of children with TBMD or MCNS had no effect. The presence of activated PLT and the effect of urine on PLT activation may be associated with the active glomerular or interstitial lesions in IgAN.

The relationship between the deposition of fibrin/fibrinogen related antigen (FRA) and morphologic findings in glomeruli was investigated in 59 children with IgA nephropathy. After treatment of renal biopsy sections with monochloroacetic acid (MCA) solution, MCA-insoluble FRA, suggesting cross-linked fibrin, was observed within the capillary walls, the mesangium and/or the sclerotic area in children with severe proliferative lesions, such as segmental sclerosis, capsular adhesions or crescent formation. Mesangial deposits of FRA were confined to electron-dense deposits, and the MCA-soluble FRA did not seem to be closely related to glomerular damage. FRA deposits within crescents may consist of MCA-soluble FRA.

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.

Various factors affecting coagulation and fibrinolysis, such as factor VIII procoagulant activity, factor VIII related antigen, von Willebrand factor activity, fibrinogen, plasminogen-plasmin, plasmin inhibitor and fibrin degradation product (F. D. P.) in blood were studied together with plasminogen activator, plasminogen-plasmin and F. D. P. in urine for a total of 45 cases of Schonlein-Henoch purpura which include 25 male and 20 female Japanese children of 2 to 15 years of age.These children were classified into 4 groups, according to clinical manifestations: Group 1, which included 20 patients with no renal complications; Group 2, which included 16 patients with acute glomerulonephritis; Group 3, which included 9 patients with nephrotic syndrome; Group 4, which included no patient in this study and in whom chronic nephritis was evident.Group 1 showed a high level of serum F. D. P. in 13 cases, at the initial stage, but the other fibrinolytic factors, factor VIII procoagulant activity, factor VIII related antigen and von Willebrand factor activity fell within normal range. The high level of serum F. D. P. returned to almost normal after 2 to 4 weeks.Group 2 showed, at the initial stage, high levels (over 150%) of factor VIII procoagulant activity, factor VIII related antigen and von Willebrand factor activity in 5-8 cases and normal ranges were reverted to 3 weeks to 3 months later. A moderate increase was observed in fibrinogen content in 8 cases and a high level of serum F. D. P. in 15. Urinary levels of plasminogen activator, plasminogen-plasmin and F. D. P. were increased in 10-20 cases. Normal ranges were reverted to 6 weeks to 3 months later.Group 3 showed, at the initial stage, high levels of factor VIII procoagulant activity, factor VIII related antigen and von Willebrand factor activity, in 6-7 cases and all levels returned to normal except for a few cases. An increase was observed in plasma fibrinogen, plasminogen-plasmin and serum F. D. P. in most cases. Levels of plasminogen activator, plasminogen-plasmin and F. D. P. in the urine were increased significantly in all cases.Renal biopsy was performed on 17 children with Schonlein-Henoch purpura accompained with renal symptoms and histological findings and deposition of fibrin or factor VIII related antigen in the glomerular lesion were investigated.Histological observations revealed with minimal change, 7 with proliferative glomerulonephritis, 2 with focal segmental glomerulonephritis and 5 with focal lobular glomerulonephritis.The degree of glomerular fibrin deposits correlated closely with plasma fibrinogen, plasminogen-plasmin in plasma and urine, and F. D. P. in serum and urine.The degree of glomerular deposition of factor VIII related antigen showed a close correlation not only with factor VIII procoagulant activity, factor VIII related antigen and von Willebrand factor activity, but with fibrin deposition.

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.

The localization of intrarenal cross-linked fibrin was examined by the effect of monochloroacetic acid treatment on the kidney sections. In acute glomerulonephritis or in mild diffuse or focal proliferative type of nephritis, cross-linked fibrin was observed mainly within glomerular capillary walls. Extension of cross-linked fibrin deposit over the mesangium or sclerotic area was seen in moderate to severe proliferative type of nephritis or in membranoproliferative glomerulonephritis. In hemolytic uremic syndrome or disseminated intravascular coagulation syndrome, cross-linked fibrin was detected within glomeruli and vessels.

Arthral, abdominal and renal symptoms in Henoch-Schonlein purpura (HSP) were scored. Coagulation factor XIII (F XIII) activity was determined in fifty-six children with HSP and the correlation with the severity score of the clinical symptoms was investigated. As a result, it was found that the decrease in F XIII level was correlated with the severity score of clinical symptoms, particularly abdominal symptoms. Based on the results, a controlled study was performed in 24 cases with moderate symptoms divided into a group treated with F XIII concentrate and a non-treated group to investigate clear-cut efficacy as a next study. In three days after the administration the symptoms were improved remarkably in accordance with the increase of F XIII level compared with non-treated group and scoring of clinical symptoms was confirmed to be useful for assessing the application of the F XIII concentrate to HSP.

This study examines the effects of anticoagulant and antithrombotic drugs on blood pressure (BP), plasma renin activity (PRA) and renal glomerular and vascular lesions in rats following aortic ligation. Whole heparin, low molecular weight (LMW) heparin or defibrinotide was administered for 4 weeks to rats following complete aortic ligation between the renal arteries. BP and PRA in whole heparin treated rats were significantly lower than those in control rats (p less than 0.01, p less than 0.01). Renal morphology also revealed a reduced number of epithelial cell droplets (ECD) (the nature of which is unknown) in glomeruli and less severe vascular lesions (p less than 0.005, p less than 0.005). Decreased BP and PRA were seen in rats receiving LMW heparin, but the failure of LMW heparin to prevent vascular damage indicates that an increased anti-Xa activity or reduction of PRA is not enough to prevent these lesions in hypertensive rats. The rats receiving defibrinotide showed reduced BP only.

3 young Japanese sisters with congenital α2-plasmin inhibitor (α2-PI) deficiency are reported. They have mild umbilical bleeding and/or repeated prolonged bleeding after minor trauma, but rarely spontaneous bleedings. The most characteristic hemostatic findings were shortened whole blood clot lysis time and euglobulin lysis time. Activities of all hemostatic factors except α2-PI were within normal range. Both functional and immunological absence of α2-PI were found in the plasma, and this failure to detect α2-PI was not corrected by the addition of the patient’s plasma of the first described case of α2-PI deficiency. Clinical and laboratory data revealed that these patients were probably homozygous for α2-PI deficiency and born of heterozygous parents, but not of consanguineous ones. Bleeding episodes due to deficiency of (α2-PI in these patients were well controlled by an antifibrinolytic agent, tranexamic acid.

1. Animals and histology from two previous investigations were used to test the hypothesis that a similar elevation in blood pressure may result in a different sequence of pathological changes in different experimental models of hypertension, DOCA-salt and aortic-ligature hypertension. 2. To asses differences in morphological parameters, individual animals in the two groups were paired (n = 12) for the same level of blood pressure at sacrifice. 3. Vascular damage was significantly less in the DOCA-salt group (P less than 0.01). Glomerular lesions however were more severe in the DOCA-salt group. In rats with aortic-ligature hypertension significantly fewer glomeruli had fibrinoid and/or crescents than in DOCA-salt rats (P less than 0.001). There were fewer glomeruli with epithelial cell droplets (ECD) and fewer ECD per glomerulus in aortic-ligature when compared with DOCA-salt hypertension (P less than 0.001, P less than 0.001, respectively). 4. This study highlights the significance of factors other than blood pressure per se in producing vessel and glomerular lesions in experimental hypertension.

Urinary levels of crosslinked fibrin degradation products (XLFDP) in glomerular disease were measured by radioimmunoassay using monoclonal antibodies. The majority of patients with crescentic glomerulonephritis or focal segmental sclerosis/hyalinosis showed increased levels of urinary XLFDP. Mild to moderately increased levels were seen in some patients with other forms of glomerulonephritis. In focal segmental sclerosis/hyalinosis or membranous nephropathy, however, XLFDP levels correlated with proteinuria and the ratio of XLFDP to proteinuria was lower than in crescentic glomerulonephritis. Although isolated urinary XLFDP is not necessarily an index of intraglomerular coagulation and fibrinolysis, the ratio of XLFDP to proteinuria may provide a useful marker for active glomerular lesions as crescents and/or intraglomerular coagulation and fibrinolysis.

The aim of the present experiments was to determine if anticoagulant and antithrombotic drugs, which protect against hypertension and vascular damage in some models of hypertension, have a similar effect in DOCA-salt hypertension. Unilaterally nephrectomized rats received injections of deoxycorticosterone acetate (DOCA) and 1% saline to drink and were treated with either heparin, defibrotide, low molecular weight (LMW) heparin or vehicle (control) for five and a half weeks. At sacrifice heparin treated rats had decreased hematocrit (p less than 0.001) and prolonged APTT (p less than 0.001). LMW heparin and defibrotide groups did not differ from control animals in either hematocrit or APTT. The systolic blood pressure (SBP) of heparin treated rats at sacrifice was lower than control (p less than 0.05) but, there were no other differences in SBP throughout the course of the experiment. Renal morphology revealed a lower number of glomerular epithelial cell droplets in the heparin group (p less than 0.01) only. Vascular damage did not differ significantly between groups.

Fibrinogen (Fg), factor VIII and factor XIII levels in the plasma were found to correlate closely with the results of renal biopsy in 29 children with renal disease, including 9 cases of acute glomerulonephritis (AGN), 9 of nephrotic syndrome (NS) and 11 of purpura nephritis (PN). At the acute phase of AGN, mild deposition of Fg, factor VIII related antigen (VIIIR: AG), factor XIII subunit A (XIII-A) and factor XIII subunit S (XIII-S) were limited to the glomerular capillaries together with a slightly increased Fg level and a low factor XIII in the plasma. In recovery or persistent cases, these values showed normal levels, and no deposition of VIII R: AG and XIII-A was detected.In NS, the Fg, factor VIII and factor XIII levels were elevated in the relapse phase, but these changes were reversed in patiants with remission. Minimal changes were not associated with detection of these coagulant factors within the glomeruli. On the other hand, intense deposition of Fg with mild VIII R: AG, XIII-A and XIII-S was detected in cases with mesangial proliferation or focal sclerosing lesions.High levels of Fg, factor VIII procoagulant activity (VIII: C) and VIII R: AG were found in the acute phase of PN, and cases with crescent formation revealed an increased level of Fg accompanied with a decrease in factor XIII. In PN, intraglomerular deposition of Fg, VIII R: AG, XIII-A and XIII-S were almost present, and a tendency to form intense or dense Fg deposits was characteristic of this disease. Deposition of Fg was observed in the crescents, but VIII R: AG, XIII-A and XIII-S deposits were not present in such lesion.

This study examines the effect of anticoagulant and antithrombotic drugs on blood pressure (BP), plasma renin activity (PRA) and renal glomerular and vascular lesions in malignant hypertensive rats. Whole-heparin, low molecular weight (LMW)-heparin, or defibrinotide were administered for 4 weeks to rats following complete aortitic legation between the renal arteries. BP and PRA in whole-heparin treated rats were significantly lower than those in control rats (p

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.

Renal biopsy tissue from 33 children with various glomerular diseases has been investigated by electron microscopy using a postembedding immunostaining technique with a protein A-gold complex in order to establish more precise correlations between immunopathologic and morphologic findings in glomeruli. This technique could detect immunoglobulins (IgG, IgA, and IgM), complement factor (C3c), and fibrinogen-related antigen. The immunoreactivity of these antigens was essentially confined to the mesangial, paramesangial, subendothelial, and subepithelial 'electron-dense deposits' in the glomeruli. Except for IgM and C3c in the case of glomerular sclerosis, the distribution of the mentioned factors was even in the electron-dense deposits, as could be shown by 'double-immunolabeling'. From the above-mentioned findings one can conclude that several of the localized factors are associated with the formation of electron-dense deposits, the ultrastructural hallmarks of glomerular disease.

Determination of coagulation Factor XIII (F XIII)-related parameters in 21 patients with Henoch-Schonlein purpura documented a significant decrease of F XIII activity as well as of the F XIII-related antigenic determinants. Subgroup analysis with regard to the clinical symptoms showed an even further decrease of these parameters in patients with gastrointestinal complications. Stimulated by these findings a substitution therapy with a F XIII concentrate was initiated in those patients whose F XIII activity in plasma remained low and who developed severe abdominal pain accompanied by persisting gastrointestinal bleeding. This therapeutic approach not only corrected the laboratory data, but more important led to a cessation of pain and bleeding. A rapid decrease of F XIII levels after transfusion below 40 U/ml was indicative of relapse of abdominal symptoms, while increasing values were associated with the recovery of the patients. In conclusion: F XIII activity determinations appear to have a predictive value in patients with Henoch-Schonlein purpura, and the administration of F XIII concentrates may contribute to the improvement of gastrointestinal complications.

Levels of antithrombin III (AT-III) activity and antigen in plasma and urine in children with renal diseases, and their correlation with the light microscopic findings of kidney tissue and the fluorescence of glomeruli, were investigated. AT-III activity in plasma was reduced slightly during the acute stage of acute glomerulonephritis and moderately in the relapse stage of nephrotic syndrome, whereas a small increase of AT-III antigen level in urine was noted in the acute stage of glomerulonephritis and considerably more was observed during the relapse stage of nephrotic syndrome. During the acute stage of glomerulonephritis or in some primary persistent glomerulonephritis (IgA nephritis, non-IgA nephritis), Henoch-Schonlein purpura nephritis and nephrotic syndrome, localization of small amounts of AT-III was noted on the capillary walls of glomeruli. These findings were in parallel with the proliferative changes of glomeruli. However, the AT-III localization did not change in parallel with the light microscopic findings or degree of the fluorescence of the fibrinogen/fibrin-related antigen. It was thought that the existence of AT-III antigen on the capillary walls of the glomeruli might be associated with the inhibition of excessive fibrin formation by AT-III.