Walentyna Zoch-Zwierz

Effect of cyclosporin A on proteinuria in the course of glomerulopathy associated with WT1 mutations

Denys–Drash syndrome (DDS) is characterized by progressive glomerulopathy caused by diffuse mesangial sclerosis (DMS), genitourinary defects, and a higher risk of developing Wilms’ tumor. It is commonly assumed that the DMS is unresponsive to any medications. In this report, we present a patient with Denys–Drash syndrome, in whom the cyclosporine A (CsA) was found to induce total remission. This observation and observations of other authors confirm that in genetic forms of nephrotic syndrome, the proteinuric effect of CsA may be due to a non-immunologic mechanism. We confirm the beneficial effect of CsA treatment in DDS; however, the potential nephrotoxicity of this drug will probably not allow long-term use.

Urinary monocyte chemoattractant protein-1 excretion in children with glomerular proteinuria

Abstract Objective. The aim of the study was to examine the urinary levels and clinical significance of monocyte chemoattractant protein-1 (uMCP-1) in children according to histological diagnosis and degree of proteinuria. Material and methods. Group I comprised 20 children with idiopathic nephrotic syndrome (INS), examined twice (A, during INS relapse; and B, after proteinuria subsided). Group II comprised 17 children with persistent proteinuria due to focal segmental glomerulosclerosis (FSGS). Group III included 12 children with immunoglobulin A nephropathy (IgAN). The control group (C) contained 22 healthy children. uMCP-1 was determined by enzyme-linked immunosorbent assay and expressed in pg/ml. Results. The median uMCP-1/creatinine ratio (uMCP-1/cr) in children with minimal change disease in relapse (IA) was significantly higher than in controls (p < 0.05), but when controlling for cyclosporine A (CsA) treatment the median uMCP-1 in children with INS, who were not treated with CsA, was 12.01 pg/mg cr (range 1.82–261.56 pg/mg cr) and did not differ from healthy controls. In examination IB the uMCP-1/cr concentration decreased and did not differ from healthy controls (p > 0.05). Children from groups II and III also had higher uMCP-1/cr levels than groups I and C (p < 0.01). uMCP-1/cr positively correlated with serum total cholesterol, low-density lipoprotein and protein/creatinine ratio in relapse (IA), and with serum cholesterol level in group B. A positive correlation between uMCP-1/cr and protein/creatinine ratio was also confirmed in groups II and III. Conclusion. Increased uMCP-1 was found in children with IgAN and FSGS correlated with proteinuria. A slight increase in uMCP-1 in children with INS was probably associated with CsA treatment.

Serum osteoprotegrin (OPG) and receptor activator of nuclear factor kappaB (RANKL) in healthy children and adolescents.

UNLABELLED Most metabolic bone diseases are characterized by a disturbance in bone resorption, therefore biochemical markers concerning this process are of special interest. Recent investigations in bone biology identified the RANKL/ RANK/OPG system, the set of cytokines or cytokine receptors belonging to the tumor necrosis factor (TNF) family that are required for control of bone modeling and remodeling. The imbalance between OPG and RANKL was found not only in pathology of bone, but also in the control of the immune and vascular systems. However, clinical application of new bone markers in children may be difficult due to lack of reference data in relation to age, sex and physiological development. AIM To investigate the relationship of serum concentrations of OPG, RANKL and OPG/RANKL ratio in relation to age, sex and parameters of physical development in healthy children and adolescents. CHILDREN AND METHODS The study was performed on a group of 70 healthy children and adolescents, divided into subgroups according to sex and age. OPG and sRANKL serum concentrations were determined using ELISA. RESULTS Serum OPG did not differ between boys and girls or younger and older children. There was no correlation between OPG level and height, weight and BMI percentiles. The level of sRANKL was 3 times higher in males than in females (p < 0.01) and almost 3 times higher in older than younger children (p < 0.01). There was a positive correlation between sRANKL concentration and body weight percentile (r = 0.268, p < 0.05). There was no correlation between serum OPG and sRANKL levels. CONCLUSION In healthy children and adolescents the serum level of OPG is not influenced by age, sex or parameters of physical development, in contrast to sRANKL and sRANKL/OPG ratio, which are dependent on these factors. Age and sex reference data should be established.

Relationship of serum interleukin-7 concentration and the coagulation state in children with nephrotic syndrome.

Background : Enhanced platelet reactivity may play a significant role in the hypercoagulable state of nephrotic syndrome (NS). Thrombocytosis with platelet aggregation cause the release of some cytokines, among them interleukin‐7 (IL‐7). The aim of the study was to evaluate serum IL‐7 levels in children with the symptoms of NS and to determine a correlation between its concentration and platelet count, other hemostatic factors, and NS intensity.

Glucocorticoid receptor and vascular endothelial growth factor in nephrotic syndrome

AIM The aim of the study was to assess plasma and urine concentrations of vascular endothelial growth factor (VEGF) in nephrotic syndrome children (NS) depending on the total dose of glucocorticoids (GC) and the percentage of lymphocytes with glucocorticoid receptor expression (CD3/GCR). METHODS We examined 51 children (2-15 years), allocated to three groups: group I: 13 children with the first NS onset, group II: 13 children with NS relapse, group C: 25 healthy children. The NS patients were examined: (A) before treatment and (B) 4-5 weeks after prednisone administration at a dose of 60 mg/m2/24 h. Plasma and urinary VEGF levels were determined using the immunoenzymatic ELISA method. Flow cytometry was applied to assess CD3/GCR expression. RESULTS Higher plasma and urinary VEGF concentrations were noted in NS children before treatment (A), as compared to control subjects (C). Following prednisone therapy (B), VEGF level was reduced but it was still higher than in the control group. Positive correlation was observed between VEGF and protein in the urine (group I r = 0.660, P < 0.05, group II r = 0.818, P<0.01) and a weak positive correlation between VEGF in plasma and urine (group I r = 0.531, P<0.05, group II - r = 0.581, P<0.05). CD3/GCR expression was lower in group II. In both groups, the correlation between plasma VEGF and CD3/GCR was positive (P<0.05). CONCLUSIONS 1. Plasma and urinary VEGF levels increase during nephrotic syndrome onset. 2. Glucocorticoid treatment reduces plasma and urinary VEGF levels in NS children.

Functional bladder capacity and urine osmolality in children with primary monosymptomatic nocturnal enuresis

Objective To assess functional day-time bladder capacity (DBC) and urine osmolality in children with primary monosymptomatic nocturnal enuresis (PMNE) according to age and sex. Material and methods A total of 263 children with PMNE were divided into two groups: Group I, 160 children (63 girls, 97 boys) aged 5–9 years (mean age 7.14±1.47 years); and Ggroup II, 103 children (25 girls, 78 boys) aged 10–15 years (mean age 12.26±1.52 years). DBC (milliliters) was the largest void of the day measured over four 24-h periods, irrespective of the diet applied. Urine osmolality was determined three times: in the evening before bed-time; at night, 2–4 h after falling asleep; and in the morning in the nocturnal void. Results DBC was smaller in Group I than in Group II (151.27 vs 199.46 ml; p<0.05). No statistically significant differences were found in relation to sex (p>0.05). The mean osmolality of the nocturnal void in the morning was 854.15 and 909.22 mOsmol/kg H2O in Groups I and II, respectively (p>0.05). Differences between boys and girls were not statistically significant (p>0.05). No correlation was found between DBC and urine osmolality (p>0.05). A detailed analysis of the results revealed DBC below the 5th percentile or above the 95th percentile in 23/263 cases (8.7%), reduced osmolality (< 800 mOsmol/kg H2O) in 76/263 (28.8%), a familial nature of nocturnal enuresis in 124/263 (47.1%) and difficulty waking in 86/263 (32.7%). Conclusions In children with PMNE aged 5–15 years, functional DBC increases with age and does not differ between the sexes; the mean nocturnal urine osmolality is neither age- nor sex-dependent.

Serum and urine leptin concentration in children with nephrotic syndrome

Literature data point to the relationship between leptin concentration and certain markers of the metabolic syndrome, including cholesterol, triglycerides and apolipoproteins. A substantial lipid metabolism disturbance occurs in children with idiopathic nephrotic syndrome (NS). The aim of the study was to find out whether in NS children, serum and urine leptin levels change proportionally to lipid metabolism disturbances. The study was performed on two groups: (I) 30 children with NS (A) before, (B) during, prednisone therapy after proteinuria regression; (II) 25 healthy children. Serum and urine leptin levels were determined by the immunoenzymatic ELISA method. Serum leptin level in NS children before and after treatment was similar to that in the control group ( p >0.05). Leptin urinary excretion in group A was approximately 60 times and in group B 24 times higher than in the controls ( p <0.01). Before treatment, children with NS had increased concentrations of TC, TG, LDL, β-lipoprotein, apolipoprotein B (apo B) ( p <0.01) and reduced HDL and apolipoprotein A (apo A) ( p <0.01). The conclusions were that: (1) in NS children leptin urinary excretion increases but its level is unchanged in serum; (2) serum leptin level is correlated with lipid parameters.

Bonn Risk Index Based Micromethod for Assessing Risk of Urinary Calcium Oxalate Stone Formation

PURPOSE The Bonn Risk Index has been used to evaluate the risk of urinary calcium oxalate stone formation. According to the original method, risk should be determined based on a 200 ml urine sample taken from a 24-hour collection. We evaluated whether the Bonn Risk Index can also be effectively determined in small urine samples. MATERIALS AND METHODS We studied 190 children and adolescents with nocturia and calcium oxalate urolithiasis. Initially Bonn Risk Index was determined according to the original method of Laube. Subsequently Bonn Risk Index was calculated using a computer program controlling a specially designed system to define the time point of induced crystallization based on consecutive urine samples of 1.5, 2.0 and 3.0 ml. RESULTS No significant differences were found in Bonn Risk Index between values obtained from 200 ml samples and those based on the micromethod with urine samples of 2 and 3 ml. CONCLUSIONS Assessment of risk of urinary calcium oxalate stone formation with Bonn Risk Index in small urine volumes, based on prototype equipment controlled by specialized computer software, is comparable to the original method. This finding facilitates the procedure and improves Bonn Risk Index determination in children.

Assessment of Lithogenic Risk in Children Based on a Morning Spot Urine Sample

PURPOSE The Bonn Risk Index has been used to evaluate the risk of urinary calcium oxalate stone formation. According to the original method, risk should be determined based on 24-hour urine collection. We studied whether the Bonn Risk Index could be measured in spot urine samples and which part of the day is most suitable for this purpose. MATERIALS AND METHODS We collected total and fractionated 24-hour urine (in a 6-hour nocturnal portion and 9 consecutive 2-hour diurnal samples) in 42 children and adolescents with calcium oxalate urolithiasis and 46 controls. Bonn Risk Index values determined from each of the urine fractions were compared to those obtained from related 24-hour urine collections. RESULTS Both groups exhibited similar circadian patterns of Bonn Risk Index values. Median Bonn Risk Index for the nighttime portion of urine in the stone group was 1.4 times higher than that obtained from the total 24-hour urine. The morning hours between 08:00 and 10:00 showed the peak lithogenic risk, and this fraction had the highest sensitivity and selectivity regarding discrimination between stone formers and healthy subjects. The afternoon hours demonstrated lower and less fluctuating crystallization risk. Despite diurnal fluctuations in Bonn Risk Index, there was still a well-defined cutoff between the groups. CONCLUSIONS Bonn Risk Index determined from urine samples collected between 08:00 and 10:00 appears optimal in separating stone formers from healthy subjects, and appears as useful as the value determined from 24-hour urine collection. Investigation of this diurnal sample simplifies diagnosis in pediatric stone disease without loss of clinical information.

Expression of multidrug resistance P-glycoprotein on lymphocytes from nephrotic children treated with cyclosporine A and ACE-inhibitor

The aim of this work was to determine the expression of P-glycoprotein (P-gp) on peripheral lymphocytes (CD3) in children with steroid-dependent nephrotic syndrome (SDNS) during cyclosporine A (CyA) and ACE-inhibitor (ACE-I) treatment. The study group (I) consisted of 20 children with SDNS aged 5–18 years, with a subsequent proteinuria relapse at the time of prednisone dose reduction. All nephrotic syndrome (NS) children were examined three times: A—at proteinuria relapse, before CyA treatment; B—after 3 months; C—after 12 months of CyA administration. The control group (II) consisted of 20 healthy children. CD3/P-gp was measured using a flow cytometry assay. The serum CyA level was assessed by means of the immunofluorescence method. The expression of CD3/P-gp in NS relapse, prior to CyA+ACE-I administration, was much higher (median 9.15%, range 1.50–13.50%) when compared to healthy controls (median 1.20%, range 0.30–5.70%). The absolute number of CD3/P-gp in this examination was almost five times higher when compared to healthy controls (p<0.01). After 3 months of CyA+ACE-I therapy, the expression of CD3/P-gp decreased dramatically and was similar to the controls. Similar results were obtained after 12 months of treatment. A strong negative correlation was found between CD3/P-gp and serum CyA concentration in both examinations (r=−0.624, p<0.01; r=−0.464, p<0.01). We conclude that the results of our studies indicate that CyA+ACE-I in SDNS inhibits the expression of P-gp. CyA is an alternative therapy that may lead to the optimization of glucocorticoid (GC) doses, thus, reducing the risk that is associated with the treatment.