Collection strategies and cryopreservation of umbilical cord blood.

The aim of this study was to compare (a) two different umbilical cord blood (UCB) collection methods while the placenta is still in the uterus (in utero), and (b) to evaluate the efficacy of four cryopreservation protocols based on UCB haematopoiestic stem cell (HSC) recovery. We analysed UCB samples collected with our original collection system designed for active Syringe/Flush/Syringe method or by standard in utero method. For comparing different cryopreservation procedures, dimethyl sulphoxide (DMSO) at final concentration of 5 and 10% was used and combined with our own controlled-rate or uncontrolled-rate cryopreservation. A total of 99 samples were collected. A significantly higher UCB volume, total nucleated cell and mononuclear cell were seen following the first collection strategy (n= 49; mean +/- SD, 103 +/- 35.4 mL; 12.34 +/- 5.27 x 10(8); 595 +/- 3.47 x 10(6)) vs. the second strategy (n= 50; 86 +/- 29.3 mL; 9.87 +/- 4.47; 424 +/- 2.82 x 10(6)) respectively (P < 0.01). The discard rate was 14% for the first and 36% for the second collection strategy (P < 0.01). It was shown that the most efficient procedure was the controlled-rate protocol combined with lower (5%) DMSO concentration. Using active Syringe/Flush/Syringe method, we collected UCB with greater volumes and with lower discard rate compared to the standard by gravity technique. The data presented also showed much better recovery of UCB cells when controlled-rate freezing procedure and 5% DMSO were combined.

Therapy of rapidly progressive glomerulonephritis with immunosuppressive agents.

Rapidly progressive glomerulonephritis is a kidney disease leading to sudden and definitive damages of the renal parenchyma and progressive impairment of its function until the complete failure. Histological findings of the changes are characterized with dominant glomerular lesions with crescentic formations. Early and intensive immunosuppressive therapy with pulse doses of steroids (Solumedrol 1.5 to 2.5 g), followed by Prednisolone 1 mg/kg every other day and cytostatic drugs (cyclophosphamide 1-2 mg-kg/every other day) discontinues the processes of specific and non-specific inflammation in the kidney and could enable regeneration of the kidney tissues with favorable outcome of the disease. Immunosuppressive therapy should be gradually lowered after two months, and applied for at least three to six months.

Shunt nephritis associated with Moraxella bovis.

Moraxella bovis was repeatedly isolated from cerebrospinal fluid in a girl with two episodes of shunt nephritis. Clinical remission of nephritis was achieved only after shunt replacement. A list of about 20 infectious agents reported to date in patients with shunt nephritis is given. M. bovis is yet another agent previously not reported in patients with shunt nephritis.

[Shunt nephritis. 2 case reports and a review of the literature]. / Santin nefritis. Prikaz dva bolesnika i pregled literature.

Two cases of glomerulonephritis associated with colonization of cerebrospinal fluid shunts inserted for the treatment of hydrocephalus (shunt nephritis) are described and additional 113 cases reported in world literature, are reviewed. Both of our patients were affected by two episodes of shunt nephritis. Complete clinical remission of renal disease followed antibiotic treatment in one patient, and shunt replacement combined with antibiotic therapy in the other. In the 115 cases of shunt nephritis reported to date, the causative organisms, clinical features, pathogenesis, renal histology, treatment, and outcome are discussed in detail. The importance of regular observation for early recognition of this reversible form of glomerulonephritis, is emphasized.

[The importance of the sonographic image in the differential diagnosis of acute rejection in kidney transplantation]. / Znacaj sonografske slike u diferencijalnoj dijagnostici akutnog odbacivanja transplantisanog bubrega.

Between October 1987 and October 1992 110 biopsies of transplanted kidney. Histologic examination showed 36 cases of acute rejection (Ac), 19 cases of chronic rejection (Chr), 16 cases of acute tubular necrosis (ATN) and 39 cases of cyclosporin nephrotoxicity. Sonographic characteristics as: hyperechogenicity of the cortex, hypoechogenicity of central pyelovascular zone, large and sonolucent pyramids, spherical configuration of renal allograft, were coelating with histologic and relevant clinic sings of disease. The results show statistically significant correlation with sonographic sings of acute rejection and relevant histologic and clinical findings. There was no correlations in other groups of diseases. These results confirm ultrasound as the primary imaging tool and an invaluable diagnostic modality in the renal transplant patient.

[Renal size as an ultrasonographic parameter in the differential diagnosis of primary and secondary glomerulopathy]. / Velicina bubrega kao ultrasonografski parametar u diferencijalnoj dijagnozi primarne i sekundarne glomerulopatije.

Between October 1987. and October 1992. 289 biopsies of the kidney in patients with glomerular diseases were performed. Pathohistologic analysis showed 218 cases with primary and 71 cases with secondary glomerular diseases. In all patients kidneys were measured by ultrasound examinations. The results showed significant difference in longitudinal diameter of kidneys in patients with primary from those with secondary glomerular diseases. These results can help a nephrologist to diagnose glomerular disease.

[Jeune's syndrome (3 case reports)]. / Zenov sindrom (prikaz tri bolesnika).

Jeune's syndrome or asphyxiating thoracic dystrophy is an autosomal recessive osteochondrodysplasia with multisystem involvement. In patients who survive neonatal period in the main clinical feature progressive renal failure is. Renal lesions are variable but is familial juvenile nephronophtisis the most frequent one. We present three patients with Jeune's syndrome phenotype and chronic tubulointerstitial disease. All patients developed terminal renal failure in the eyrly childhood. Renal histology, examined in two cases, was consistent with juvenile nephronophtisis in one case and with renal dysplasia in other case. All our patients had hepatic fibrosis and two of them had pigmentary retinophaty. We want to underline the importance of regular check-up of children with typical phenotype by pediatrician-nephrologist as wel as possibility of prenatal diagnosis of Jeune's syndrome.

[Idiopathic membranoproliferative glomerulonephritis (structural-functional relationship)]. / Idiopatski membranoproliferacioni glomerulonefritisi (strukturno-funkciona zavisnost).

We investigated 12 patents with idiopathic membranoproliferative glomerulonephritis. Discriminatory analysis was used for structural functional relationship in order to determine discriminatory power of certain clinical and pathohistological parameters. Semiquantitatively were determined pathohistological parameters: glomerular index in range 0-10, vascular index 1-4, interstitial fibrosis 0-10, interstitial infiltration 0-3, tubular atrophy 0-3. Nephrotic syndrome (symbolic value), quantitative proteinuria and creatinine clearance (continual variable) were used as clinical parameters. Discriminatory power was determined as a degree of decreasing Shennon's entropy dy distinction of patients according to value of creatinine clearance at the time of biopsy. Discriminatory power was measured in the information measurement units (bit). The most powerful was glomerular index (discriminatory power 0.29) in moderately reduced glomerular filtration rate (creatinine clearance 80 ml/min) at the time of biopsy. In severely reduced glomerular filtration rate (creatinine clearance 40 ml/min) vascular index had the greatest discriminatory power (0.24) while interstitial infiltration and interstitial fibrosis had less powerful discriminatory power (0.13). Negative predictive value of reduced glomerular filtration rate at the time of biopsy in membranoproliferative glomerulonephritis has been confirmed in other studies.

[Renal function and histologic findings in immunoglobulin A nephropathy]. / Funcija bubrega i histoloski nalaz u nefropatiji imunoglbulina A.

We investigated 28 patients with IgA nephropathy. Discriminatory analysis was the method used to determine discriminatory power of certain clinical and pathohistological parameters. Semiquantitatively were determined pathohistological parameters: glomerular index in range 0-10, vascular index 1-4, interstitial fibrosis 0-10, interstitial infiltration 0-3, tubular atrophy 0-3. Nephrotic syndrome (symbolic value), quantitative proteinuria and creatinine clearance at the time of biopsy and year after (continual variable) were used as clinical parameters. Discriminatory power was determined as a degree of decreasing Shennon's entropy by distinction of patients according to value of creatinine clearance at the time of biopsy, and a year after biopsy. Discriminatory power was measured in he information measurement units (bit). Based on discriminatory-predictive analysis we determined that glomerula changes and interstitial fibrosis had an equal effect (discriminatory power) on renal function at the time of biopsy and a year latter, as well as predictive value of these histological parameters and creatinine clearance at the time of biopsy.

[Glutathione and enzymes associated with glutathione metabolism in adriamycin nephropathy]. / Glutation i kljucni enzimi metabolizma glutationa u adriamicinskoj nefropatiji.

Recent data have shown the protective effect of thiol groups on the progression of adriamycin-induced experimental nephropathy, in which reactive oxygen metabolites have been postulated to play an important role. To gain greater insight in the role of glutathione (GSH) in adriamycin-induced nephrosis, we studied changes in reduced GSH level and its associated enzymes in kidney tissue of rats undergoing chronic renal failure produced by i.v. infections of adriamycin (2 x 2 mg/kg b.w.). Kidney damage was characterized by increases in relative kidney weight and BUN levels. The results obtained revealed a 15% drop in renal GSH level in adriamycin-treated animals associated by a similar decrease in the gamma-glutamylcysteine synthetase activity. The activities of kidney glutathione reductase (GR) and glutathione peroxiase (GSH-Px) which are critical constituents of GSH-redox cycle, were significantly decreased (23 an 26%, respectively) in response to adriamycin treatment. Rat kidney glutathione-S transferase and gamma-glutamyl transpeptidase activities were not affected in adriamycin induced nephrosis. We propose that the impairment of renal antioxidant defense, characterized by combined drop in GSH, GR and GSH-Px levels, could permit enhanced free radical induced kidney damage in adriamycin-induced nephropathy.

[Cyclosporine in the treatment of primary focal-segmental glomerulosclerosis]. / Ciklosporin u lecenju primarne fokusno-segmentne glomeruloskleroze.

Cyclosporin A (CyA) was administered at the Clinic of Nephrology, CCS, in treatment of 8 patients with steroid-resistant nephrotic syndrome (NS) caused by focal segmental glomerulosclerosis (FSG), Cyclophosphamide (CP) treatment was attempted in 6 of these cases, but without success. Prior to the onset of CyA therapy, CP treatment was interrupted, and the dose of prednisone was reduced to 0.4 mg/kg/48 h. CyA was applied in the initial dose of 5 mg/kg/bw/24h, and then adjusted so that CyA blood level was 80-120 ng/ml. Although CyA treatment caused proteinuric decrease in most of the patients (from 12.1 g/24 h to 8.2 g/24 h), complete NS remission was achieved in only one patient, the same as the incomplete one. The female patient with a positive response to CyA treatment behaved as "CyA-dependent" and each discontinuation of CyA therapy soon led to recurrent NS. Insignificant CyA effect on NS or renal function was registered in 4 patients, and the therapy was interrupted in two cases due to renal function impairment. Apart from nephrotoxicity, other CyA side effects were absent.

[Acute tubulointerstitial nephritis with uveitis]. / Akutni tubulointersticijumski nefritis s uveitisom.

Clinical features and natural course of acute tubulointerstial nephritis and uveitis (TINU syndrome) in five adolescent patients (3 girls and 2 boys), are presented. Initial nonspecific symptoms preceding nephropathy were anorexia, weight loss, fever and malaise. Inflammatory syndrome consisted of increased ESR, high plasma proteins and gamma globulins. Analysis of urine showed proteinuria and sterile leukocyturia. Laboratory features of tubular dysfunction and decreased GFR were found in all patients. Renal biopsy, which was performed in 2 pts, revealed acute interstitial nephritis. Anterior uveitis which appeared later, was successfully treated with topical steroids. Renal function completely recovered within a few month in four pts and markedly improved in one. Despite the fact that renal biopsy was not performed in all children, the combination of an acute nonoliguric renal failure without hypertension and signs of tubular dysfunction together with particular benign course, suggested acute idiopathic TINU syndrome.

[Lupus nephritis--clinical and morfological correlation]. / Lupus-nefritis (klinicko-morfoloska korelacija).

During the last five years at our Department renal biopsy was carried out in 41 patients with systemic lupus erythematosus. On the basis of the pathologic findings glomerular changes were classified as follows: minimal in 5 patients; mesangial proliferative glomerulonephritis in 11; focal and segmental proliferative glomerulonephritis in 5; diffuse proliferative glomerulonephritis in 15; membranous glomerulonephritis in 3, and diffuse sclerosing glomerulo-nephritis in 2 cases. Morphological changes of renal biopsy did not correlate with the degree of deterioration of glomerular function. Twenty seven patients were regularly followed-up. Most of them had diffuse proliferative glomerulonephritis (11) and mesangial proliferative glomerulonephritis (7). This is a larger number than in other authors' series. The clinical manifestation of lupus nephritis was: nephrotic syndrome (19 patients); minimal urinary abnormalities (6); rapidly progressive renal failure (1), and acute nephrotic syndrome (1). The majority of patients had high blood pressure (23). Their immunosuppressive therapy was ordered according to renal histologic lesions and severity of extrarenal manifestations. Clinical remission was achieved in 22 patients, but 2 died due to extrarenal complications.

[Lupus nephritis in children and adolescents (Part II: Prognosis)]. / Lupusni nefritis kod dece i adolesenata (prognoza bolesti).

In 27 children and adolescents (24 females, 3 males suffering from lupus nephritis the important role of renal pathohistologic changes in the evolution and outcome of the disease was analysed, and a five-year survival rate was calculated. The mean age at the onset of the disease was 12.1 +/- 1.9 (range: 0.7-0) years, the mean interval from onset to renal biopsy was 2.1 +/- 1.9 (0.1 - 7.0) years, ant the mean follow-up was 4.1 +/- 2.1 (1-8) years. The pathohistologic changes were classified according to the World Health Organisation criteria. At the end of the follow-up patients classified in Ib and II classes were without urinary abnormalities, i.e. without preoteinuria and/or haematuria. However, patients in class IV manifests nephrotic syndrome (2 pts), renal failure (3 pts) or proteinuria and heamaturia (6 pts), and two patients died. More severe clinical features and course of lupus nephiritis associated with infavourable outcome significantly correlated with WHO classes III and IV. A five-year survival rate (life-table method) of 88.5% is among the best results reported to date in children and adolescents with lupus nephritis.

[Lupus nephritis in children and adolescents (clinical-morphologic correlation)]. / Lupusni nefritis kod dece i adolescenata (klinicko-morfoloska korelacija).

Clinical and laboratory features in 27 children and adolescents (24 females, 3 males) with lupus nephritis were analysed and correlated with pathohistologic findings obtained by renal biopsy. The mean age at the onset of the disease was 12.1 +/- 1.9 (range 0.1-7.0) years. In the 30% of patients nephritis was one of the presenting features of the disease. The most frequent signs of the lupus nephritis at the time of biopsy were proteinuria (9 pts), proteinuria and haematuria (9 pts), and nephrotic syndrome (8 pts), while 7 patients showed decreased glomerular filtration rate (GFR), and hypertension was present in five. Distribution of patients according to the WHO morphologic classification of Lupus nephritis was as follows: 6 pts showed class Ib, 6 class II, 2 class III and 13 class IV. More severe renal dysfunction, manifested by nephrotic syndrome, proteinuria with haematuria, decreased GFR and hypertension was significantly associated with proliferative lupus nephritis (combined WHO classes III and IV). Nephrotic syndrome, haematuria and decreased serum C3 with significant probability predicted proliferative lupus nephritis, while decreased GFR was of borderline predictive value.

Progressive tubulointerstitial nephritis and chronic cholestatic liver disease.

We report the clinical and morphological features of a distinctive hepatorenal disorder in four patients and review the five similar patients in the literature. The main clinical characteristics were early onset of cholestatic liver disease and progressive tubulointerstitial nephritis leading to renal death in early childhood. Liver histology showed disturbed architecture with nodular and acinar formations and portal fibrosis and bile duct proliferation. Histological abnormalities in the kidney were severe interstitial fibrosis and tubular atrophy and dilatation, while the typical features of nephronophthisis were lacking. These clinical and morphological characteristics distinguish our patients from the majority described, as having nephronophthisis and congenital hepatic fibrosis or any other known syndrome with concomitant hepatorenal involvement. We suggest that the association of cholestatic liver disease and progressive tubulointerstitial nephritis represents a new syndrome.

Laminin, fibronectin, and Goodpasture antigen detection in patients with Alport's syndrome.

Indirect immunofluorescence study with laminin and fibronectin monoclonal antibodies on paraffin sections, as well as with serum from a patient with Goodpasture's syndrome with high titer of autoantibodies that recognize the antigenic determinants in human glomerular and tubular basement membrane, was performed on 14 patients with Alport's syndrome and 5 specimens of normal renal tissue obtained from donors in cases of renal transplantation (control group). We found no binding of Goodpasture antigen to glomerular and distal tubular basement membranes in renal biopsy tissue from all 14 patients with Alport's syndrome. In contrast, there was bright linear fluorescence of Goodpasture antigen on glomerular and tubular basement membranes of normal renal material. There was no difference in laminin and fibronectin binding in patients with Alport's syndrome and controls. In all the cases binding was strongly positive. These results suggest an abnormality or absence of immunoreactive autoantigen in the glomerular and distal tubular basement membrane in patients with Alport's syndrome. Therefore, Goodpasture antigen detection could be an important diagnostic method in early stages of Alport's syndrome when characteristic morphological changes are not yet developed.

Spontaneous blastogenesis profiles and IL-2 receptor expression on kidney allograft.

To examine immune activation rate of interstitial and glomeruli infiltrating MNC in different conditions of human renal allograft function deterioration, 33 renal transplant biopsies were performed 1-30 months after transplantations. Forty-one patients observed were on immunosuppressives: Pr, Aza, CsA following renal transplantation from a living-related donor parent. The patients were divided according to their histologic diagnosis into the following groups: 1, 15 pts in acute rejection attack (AR); 2, 10 pts with cyclosporine nephrotoxicity (CsN); 3, 10 pts with chronic vascular rejecting kidney (ChR). A conventional histologic investigation and immunohistochemical analyses of CD3 and CD25 molecules were performed in groups 1-3. Spontaneous blastogenesis (SB) of peripheral lymphocytes was simultaneously determined and compared with the controls (C)-30 healthy people, and with patients with stable renal allograft function (S)-8 pts. The highest IL-2R expression on diffuse or focal dense MNC infiltrates in interstitium was observed during AR, without IL-2R+ MNC in glomeruli. Low to moderate focal interstitial infiltrates in damaged areas of renal parenchyma due to CsN, were IL-2R negative. In ChR, moderate IL-2R expression was observed on interstitial spare mild or focal dense MNC infiltrates with IL-2R expression present on glomeruli infiltrating MNC. Significant increases of SB values were recorded during the first week after transplantation and AR in comparison to C. The highest SB values were in ChR group. Values of SB in CsN and S were on the C and before transplantation levels.(ABSTRACT TRUNCATED AT 250 WORDS)