Serum and urine periostin and cytokeratin-18 in children with congenital obstructive nephropathy.

Congenital obstructive nephropathy (CON) is one of the most common causes of chronic kidney disease in children. The aim of the study was to investigate serum and urine periostin and cytokeratin-18 (CK-18) in children with CON in relation to CON etiology, treatment, and kidney injury. We evaluated 81 children with CON secondary to ureteropelvic junction obstruction (UPJO), ureterovesical junction obstruction (UVJO), posterior urethral valves (PUV) and 60 controls. Neither biomarker demonstrated any relation to CON etiology. However, all patients showed significantly higher urine periostin (uPeriostin) and uPeriostin/Cr levels than the controls. Also, UVJO patients showed higher sCK-18 and uCK-18/Cr levels, and PUV patients showed higher uCK-18/Cr levels than the controls. Neither biomarker was found to have any relation to CON treatment. However, conservatively treated children and those before and after surgery showed significantly higher uPeriostin and uPeriostin/Cr levels than the controls. uPeriostin strongly correlated with differential renal function (DRF) < 40%. The ROC analysis demonstrated the best area under the curve (AUC) for uPeriostin (0.831) and uPeriostin/Cr (0.768), and low for sPeriostin (0.656) and uCK-18 (0.615) for detecting renal injury. In conclusion, although serum and urine periostin and CK-18 did not display any relation to etiology or the type of CON treatment, uPeriostin seems to be a useful tool for detecting renal injury in children with CON, especially due to its strong negative correlation with DRF < 40%.

Urinary vanin-1 for predicting acute pyelonephritis in young children with urinary tract infection: a pilot study.

BACKGROUND: Vanin-1, an epithelial glycosylphosphatidylolinositol (GPI)-anchored pantetheinase, is a valuable marker of renal injury. PURPOSE: The aim of this study was to assess the predictive value of vanin-1 in acute pyelonephritis (APN) in comparison to the conventional serum inflammatory markers in children aged 1-24 months with the first episode of urinary tract infection (UTI). MATERIAL AND METHODS: Urinary vanin-1, vanin-1/Cr ratio, WBC, CRP, PCT were analysed in 58 children with febrile UTI and in 18 children with non-febrile UTI. Febrile UTI group was divided into APN subgroup (n = 29) and non-APN subgroup (n = 29), based on the results of Tc-99m-ethylenedicysteine scan. RESULTS: The mean vanin-1 level was higher in the APN group compared to the non-febrile UTI group (p = 0.02) and did not differ between APN and non-APN subgroup. In univariate analysis, vanin-1 (p = 0.042), CRP (p < 0.001), PCT (p < 0.001), and WBC (p = 0.022), were associated with APN, but only vanin-1 (p = 0.048) and CRP (p = 0.002) were independent markers of APN. In ROC analysis, vanin-1, with its best cut-off value of 16.53 ng/mL, had worse diagnostic profile (AUC 0.629, sensitivity 58,6%, specificity 63.8%) than CRP, PCT and WBC (AUC: 0.937; 0.880; 0.667, respectively). CONCLUSIONS: Vanin-1 is not useful for predicting APN, since its diagnostic value is inferior to other conventional serum inflammatory markers.

Serum Klotho is correlated to cardiovascular complications of chronic kidney disease in children.

AIM: Aim of the study was to investigate soluble Klotho (sKl), fibroblast growth factor 23 (FGF23) concentrations, and their correlations with cardiovascular complications in children with CKD. MATERIALS AND METHODS: 38 children with CKD stages 2 - 5 were compared to 38 healthy controls in terms of: plasma FGF23, serum sKl, peripheral and central blood pressure, arterial stiffness (pulse wave velocity - (PWV)), carotid intima media thickness (cIMT), left ventricular mass index (LVMI), and diastolic function. Correlations between FGF23, sKl, and cardiovascular parameters were investigated. RESULTS: The CKD group was characterized by higher FGF23, lower sKl concentrations, higher peripheral and central blood pressure, arterial stiffness, cIMT, left ventricular mass index, and decreased E/A ratio compared to the control group. In CKD children, sKl correlated negatively with diastolic blood pressure (DBP), mean arterial pressure (MAP), central systolic, diastolic, and mean blood pressure, PWV, and LVMI. In multivariate analysis, higher sKl was a significant predictor of lower peripheral and central DBP and lower LVMI and E/A, whereas higher FGF23 was a predictor of higher of LVMI. CONCLUSION: (1) In children with CKD, decreased sKl might be a marker of elevated central blood pressure. (2) Both sKl decrease and FGF23 increase could possibly contribute to left ventricular hypertrophy in this group of patients.

Renalase in children with chronic kidney disease.

Background: Renalase is kidney-derived molecule initially considered as catecholamine-inactivating enzyme. However, recent studies suggest that renalase exerts potent cardio- and nephroprotective actions, not related to its enzymatic activity. Purpose: To assess renalase level in children with chronic kidney disease (CKD). Material and methods: Serum renalase, BMI, arterial stiffness, peripheral and central blood pressure, intima-media thickness (IMT), medications, and biochemical parameters were analyzed in 38 children with CKD (12.23 ± 4.19 years) (stage G2-5). Control group consisted of 38 healthy children. Results: In the study group, GFR was 25.74 ± 8.94 mL/min/1.73 m2; 6 children were dialyzed; 26 had arterial hypertension. Renalase level was higher in the study group compared to control group (p < 0.001). In CKD children renalase correlated (p < 0.05) with BMI Z-score (r = -0.36), alfacalcidol dose (r = 0.41), GFR (r = -0.69), hemoglobin (r = -0.48), total cholesterol (r = 0.35), LDL-cholesterol (r = 0.36), triglycerides (r = 0.52), phosphate (r = 0.35), calcium-phosphorus product (r = 0.35), parathormone (r = 0.58), and pulse wave velocity Z-score (r = 0.42). In multivariate analysis GFR (ß = -0.63, p < 0.001), triglycerides (ß = 0.59, p = 0.002), and alfacalcidol dose (ß = -0.49, p = 0.010) were determinants of renalase. Conclusions: In children with CKD there is a strong correlation between renalase level and CKD stage. Furthermore, in these patients renalase does not correlate with blood pressure but may be a marker of arterial stiffness.

Disturbances in NK Cells in Various Types of Hemophagocytic Lymphohistiocytosis in a Population of Polish Children.

Hemophagocytic lymphohistiocytosis (HLH) is a life-threatening disease associated with immune system hyperactivation and the appearance of serious systemic disturbances. The purpose of this study was an assessment of natural killer (NK) cell disturbances in a group of children with clinical signs of HLH. A total of 43 children with HLH and 17 healthy children were enrolled in the study. NK phenotyping, intracellular perforin staining, and cytotoxicity tests were performed by using the flow cytometry method. HLH patients were divided into 6 HLH types: 9% infection-related HLH; 7% malignancy-related HLH; 21% macrophage activating syndrome; 12% familial hemophagocytic lymphohistiocytosis; 2% X-linked lymphoproliferative syndrome; and 49% as HLH of unknown background. A positive correlation was observed between cytotoxicity and NK cells in children with HLH (P=0.01). In all HLH groups, the percentage of NK cells was significantly lower than in the control population. The spontaneous cytotoxicity was significantly lower in HLH patients. The results presented in this study indicate the importance of impaired function and the number of NK cells in the pathogenesis of HLH. Nonetheless, the background of disturbances seems to be different in various cases.

Markers of endothelial injury and subclinical inflammation in children and adolescents with primary hypertension.

INTRODUCTION: Adhesion molecules: E-selectin and intercellular adhesion molecule 1 (ICAM-1) are well-established markers of endothelial injury. The aim of the study was to assess the relation between E-selectin and ICAM-1 and clinical and biochemical parameters in children and adolescents with primary hypertension (PH). MATERIAL AND METHODS: In 77 patients with PH (15.04 ±2.62 years, 50 boys, 27 girls) we evaluated serum E-selectin, ICAM-1, and selected clinical and biochemical parameters including inflammatory indicators and ambulatory blood pressure monitoring (ABPM). RESULTS: The E-selectin concentration was 55.63 ±26.49 ng/ml and the ICAM-1 concentration was 302.17 ±67.14 ng/ml. E-selectin and ICAM-1 correlated (p < 0.05) with BMI Z-score (r = 0.24, r = 0.29), ICAM-1 also with uric acid (r = 0.35), HDL-cholesterol (r = -0.28), platelet-to-lymphocyte ratio (r = 0.26), and systolic and mean blood pressure variability (r = 0.24, r = 0.24); in boys ICAM-1 correlated with mean platelet volume (r = 0.29). In multivariate analysis the only significant predictor of E-selectin was mean arterial pressure during 24 hours (ß = 0.329, 95% CI: 0.012-0.646) and of ICAM-1 - uric acid (ß = 0.430, 95% CI: 0.040-0.819). In 27 children with newly diagnosed PH E-selectin correlated negatively with diastolic blood pressure dipping (r = -0.54, p = 0.004) and positively with ambulatory arterial stiffness index (r = 0.51, p = 0.012). CONCLUSIONS: Elevated mean arterial pressure and hyperuricemia are risk factors of endothelial damage in paediatric patients with primary hypertension. In children with untreated primary hypertension there may be a relation between endothelial damage and disturbed circadian blood pressure profile.

Asymmetric dimethylarginine is not a marker of arterial damage in children with glomerular kidney diseases.

INTRODUCTION: Asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide synthase, correlates with cardiovascular risk especially in patients with chronic kidney disease. The aim of our study was to establish significance of ADMA as a biomarker of arterial damage in children with glomerulopathies. MATERIAL AND METHODS: In 80 children with glomerulopathies (mean age, 11.33 ±4.25 years; 42 with idiopathic nephrotic syndrome [INS], 38 with IgA or Henoch-Schoenlein nephropathy [IgAN/HSN]), we analyzed serum ADMA [nmol/ml], peripheral and central blood pressure, arterial stiffness (augmentation index - AIx75HR, pulse wave velocity - PWV), common carotid artery intima media thickness (cIMT), and selected clinical and biochemical parameters. RESULTS: In the study group, mean ADMA concentration was 1.66 ±1.19 [nmol/ml] and did not differ between INS and IgAN/HSN patients. We found no significant correlations between concentration of ADMA, cIMT [mm]/Z-score, PWV [m/s]/Z-score, and AIx75HR [%] in the whole group and in INS and IgAN/HSN patients. In the whole group of 80 children, ADMA correlated (p < 0.05) with BMI Z-score (r = -0.24), uric acid (r = -0.23), HDL-cholesterol (r = -0.25), and central mean arterial pressure (r = -0.25), in children with INS also with total protein (r = 0.37), albumin (r = 0.36), and total cholesterol (r = -0.40, p = 0.028). In multivariate analysis, serum albumin was the strongest determinant of ADMA in the whole group (ß = 0.536, 95% CI: 0.013-1.060, p = 0.045). CONCLUSIONS: 1. In children with glomerulonephritis, measurement of asymmetric dimethylarginine cannot replace well established and validated methods of assessment of subclinical arterial damage. 2. In children with glomerular kidney diseases, ADMA concentration is related primarily to serum albumin concentration.

Chemotherapy-Induced Takotsubo Syndrome.

Cardiovascular complications are a significant problem in systemically treated cancer patients. One such complication is Takotsubo cardiomyopathy, also known as Takotsubo syndrome. It is most frequently defined as a sudden and transient left or right ventricular systolic dysfunction; mimicking acute coronary syndrome, but without the associated changes in coronary arteries. Takotsubo syndrome is a relatively little known complication that appears in the course of oncological treatment, and its incidence has not yet been established. In this study, we reviewed Medline database according to case reports concerning takotsubo syndrome appearing after systemic treatment in oncological patients. We took into consideration all types of anticancer drugs. We reviewed the changes reported in the electrocardiography, echocardiography, and coronary angiography, and also the level of troponin, a marker of acute coronary syndrome elevation. In view of the increasing frequency of cardiac complications reported in patients receiving systemic oncological treatment, Takotsubo syndrome appears to be underdiagnosed. However, the syndrome may be linked to potentially fatal complications such as cardiogenic shock or cardiac arrest. Therefore, it seems essential to carry out appropriate diagnostic procedures for every patient experiencing clinical side effects of onco-pharmacotherapy. In patients with chest pain and dyspnea during or after treatment, Takotsubo syndrome should be considered, particularly that the syndrome requires a different therapy approach than that used in a coronary syndrome. Diagnostic procedures should include echocardiogram and the assessment of myocardial necrosis markers and natriuretic peptides.

[Copeptin in children with chronic kidney disease]. / Kopeptyna u dzieci chorych na przewlekla chorobe nerek.

Studies in adult patients suggest that copeptin (C-terminal fragment of antidiuretin propeptide) is related to kidney and cardiovascular diseases. AIM: The aim was to assess copeptin concentration in children with chronic kidney disease (CKD). MATERIALS AND METHODS: In a group of 38 children with CKD (age: from 4.70 to 18.00 mean 12.23±4.19 years) we evaluated: serum copeptin concentration [ng/mL], age, sex, etiology of CKD, presence of arterial hypertension (AH), medications, glomerular filtration rate (GFR), hemoglobin, calcium-phosphorus metabolism parameters, and lipids. Control group consisted of 38 healthy children aged from 5.51 to 18.0 mean 11.79±3.29 years. RESULTS: Serum copeptin concentration did not differ between children with CKD and healthy children (0.72±0.34 vs. 0.84±0.33 [ng/mL], p=0.088). In children with CKD there were no differences in copeptin concentration depending on sex, presence of AH, and CKD grade. In children with CKD only positive correlation between copeptin and hemoglobin concentrations was found (r=0.35, p=0.031); no other significant correlations between copeptin and clinical and biochemical parameters including GFR were revealed. Also no significant correlations were found between copeptin and evaluated parameters in the control group. CONCLUSIONS: In children copeptin concentration does not seem to be related to kidney function. Copeptin may be a marker of hydration status in children with chronic kidney disease. There is a need for further studies evaluating clinical significance of copeptin in children with chronic kidney disease.

Serum GDIgA1 levels in children with IgA nephropathy and Henoch-Schönlein nephritis.

INTRODUCTION: GDIgA1 (galactose deficient IgA1) plays a significant role in the pathogenesis of IgA nephropathy (IgAN) and Henoch-Schönlein nephritis (HSN). AIM OF THE STUDY: The aim of this study was to assess the relevance of serum GDIgA1 level as a prognostic marker in children with IgAN and HSN. MATERIAL AND METHODS: 41 children were included to the study group (15 IgAN, 26 HSN) and 22 to the control group. The following parameters were evaluated at baseline and endpoint: proteinuria, erythrocyturia, serum creatinine, serum IgA, GFR. A kidney biopsy was performed in all patients and evaluated according to the Oxford Classification (1 - present, 0 - absent: M - mesangial hypercellularity; E- endocapillary hypercellularity; S - segmental sclerosis/adhesion; T - tubular atrophy/interstitial fibrosis), and was calculated as the total score (sum of M, E, S, T). At the end of follow-up, the serum GDIgA1 concentration was measured. RESULTS: The serum GDIgA1 concentration in patients with IgAN and HSN was significantly higher than in the control group. No significant differences in mean proteinuria, erythrocyturia, GFR, MEST score, or GDIgA1 in serum, as well as the duration of follow-up between IgAN and HSN were observed. Baseline serum IgA concentration and time to kidney biopsy were significantly higher in children with IgAN than in children with HSN. We observed a positive correlation between GDIgA1 and IgA levels (r = 0.53), and GDIgA1 and serum creatinine levels (r = 0.5), as well as negative correlation between GDIgA1 and GFR (r = -0.37). CONCLUSIONS: Serum GDIgA1 level may have a prognostic value in children with IgAN and HSN; however, to fully elucidate its clinical potential further studies performed in larger patient cohorts are required.

Renalase in Children with Glomerular Kidney Diseases.

Studies suggest that renalase, a renal catecholamine-inactivating enzyme, plays a major role in the pathogenesis of kidney and cardiovascular diseases in adults. This study seeks to determine the role of renalase in children with glomerular kidney diseases. We evaluated the serum renalase, arterial stiffness, intima-media thickness, blood pressure, and clinical and biochemical parameters in 78 children (11.9 ± 4.6 years of age) with glomerulopathies such as idiopathic nephrotic syndrome (40 cases), IgA nephropathy (12 cases), Henoch-Schönlein nephropathy (12 cases), and other glomerulopathies (14 cases). The control group consisted of 38 healthy children aged 11.8 ± 3.3 years. The mean renalase was 25.74 ± 8.94 µg/mL in the glomerulopathy group, which was not significantly different from the 27.22 ± 5.15 in the control group. The renalase level did not differ among various glomerulopathies either. However, proteinuric patients had a higher renalase level than those without proteinuria (28.43 ± 11.71 vs. 24.05 ± 6.23, respectively; p = 0.03). In proteinuric patients, renalase correlated with daily proteinuria. In the entire glomerulopathy group, renalase correlated with age, systolic central blood pressure (BP), diastolic peripheral and central BP, mean peripheral and central BP; peripheral diastolic BP Z-score, glomerular filtration rate, cholesterol, triglycerides, and pulse wave velocity. We conclude that in children with glomerulopathies renalase, although basically not enhanced, may underlie blood pressure elevation and arterial damage.

Effectiveness of rituximab in nephrotic syndrome treatment.

Idiopathic nephrotic syndrome (INS) is a common chronic illness characterized by massive proteinuria and hypo-albuminemia in children. Baseline treatment is 6 month-corticotherapy. In cases of steroid resistant/dependent INS several types of treatment are used, including course of methyloprednisolone "pulses", alkylating agents, cyclosporin A, levamisole and mycophenolate mofetil. It has been suggested that children with frequently relapsing nephrotic syndrome or steroid-dependent nephrotic syndrome had a significantly longer relapse-free period if rituximab (RTX) treatment was additionally applied. We present a case of a 4.5 boy who due to steroid-sensitive, steroid-dependent nephrotic syndrome has been successfully treated with RTX. Administration of the one dose of Rituximab in the patient caused immediate decrease of CD19/CD20 positive B lymphocyte population. The depletion of B cells has been observed for the next six months. With regard to the fact that RTX treatment may affect patient's immune response, comprehensive immunodiagnostic has been conducted in a course of the Therapy.

Usefulness of urinary collagen IV excretion for predicting the severity of Henoch-Schönlein nephropathy children.

THE AIM OF THE STUDY: The aim of the study was to evaluate the usefulness of urinary collagen IV (Col IV) excretion for predicting the severity of autoimmune renal inflammation in children with HSN (Henoch-Schönlein nephritis). MATERIAL AND METHODS: We studied 26 children, in whom HSN was diagnosed based on kidney biopsy. In all patients, urinalysis was performed and 24-hour urinary protein excretion was measured at the onset of the disease. All kidney biopsies were also scored using the Oxford classification: M - mesangial hypercellularity score (M0 absent, M1 present); E - presence of endocapillary proliferation (E0 absent, E1 present), S - segmental glomerulosclerosis/adhesion (S0 absent, S1 present), T - tubular atrophy/interstitial fibrosis (T0 ≤ 25%, T1 26-50%, T2 > 50%). The MEST score was calculated as the sum of M + E + S + T. RESULTS: Urinary Col IV level was significantly higher in the study group than in control group. Urinary Col IV level was insignificantly higher in group A (nephrotic proteinuria) compared to the B (non-nephrotic proteinuria) and C (without proteinuria).We found no significant differences in the age at the disease onset, severity of proteinuria, and Col IV between groups 1 (S0, T0) and 2.(S1,T1/T2). The MEST score was significantly higher in group 2 than group 1. CONCLUSIONS: Urinary Col IV excretion in children with HSN may be related to the lesions severity by the Oxford classification but seems to be associated with the mean value (the MEST score). In younger children, a more aggressive disease course is observed, and thus earlier and more aggressive treatment should be considered in this group.

Effect of perinatal risk factors on neutrophil gelatinase-associated lipocalin (NGAL) level in umbilical and peripheral blood in neonates.

INTRODUCTION: Acute kidney injury biomarkers are opening a new era in diagnosing kidney failure. The requirement for a specific and sensitive marker of kidney function is highly desirable in neonates because the diagnostic possibilities in this age group are not sufficient. Recent research show that neutrophil gelatinase-associated lipocalin (NGAL) can have a great potential but there is a wide range of medical conditions, that may influence their expression. THE AIM OF THE STUDY: was to evaluate the impact of perinatal risk factors on NGAL level in neonates. MATERIAL AND METHODS: NGAL was measured in umbilical cord blood and peripheral blood in full term neonates with perinatal risk factors during the first days of life. RESULTS: We found significantly higher umbilical cord blood NGAL levels in neonates with perinatal risk factors (117.69 ng/ml) compared to the control group (64.37 ng/ml). No significant difference in peripheral blood NGAL level was shown between the two groups. Umbilical cord blood NGAL level correlated positively with peripheral blood NGAL level (r = 0.36, p < 0.01). Umbilical cord blood NGAL level was significantly higher in neonates with fetal distress and infection compared to neonates with other perinatal risk factors. Peripheral blood NGAL level was significantly higher in neonates with infection compared to neonates with other perinatal risk factors. Significantly higher umbilical cord blood NGAL levels were seen in neonates born by operative delivery compared to born by natural delivery.

Markers of bone metabolism in children with nephrotic syndrome treated with corticosteroids.

The aim of the study was to assess bone mineral density, bone metabolism markers, and vitamin D level in children with idiopathic nephrotic syndrome in the course of 1-year observation. Twenty five children with nephrotic syndrome aged 5-17 years were enrolled into the study. The median number of relapses was 6 (range 1-22). All patients were treated with prednisone and vitamin D (800 IU/day). Bone mineral density of total body (TB-BMD) and lumbar spine (L-BMD), evaluated by dual energy X-ray absorptiometry (DXA) expressed as Z-score, and serum calcium, phosphorus, parathormone (iPTH), alkaline phosphatase (ALP), bone alkaline phosphatase (BAP), osteocalcin (OC), albumin, creatinine, 25(OH)D3, 1,25(OH)2D3 and urine calcium/creatinine ratio (uCa/Cr) were evaluated at the enrollment visit and after 1 year of therapy. After 1 year significant decreases of TB-BMD Z-score (from -0.24±1.34 to -0.74±1.31, p<0.05) and 25(OH)D3 serum level (from 31.7±16.3 to 23.7±9.3; p<0.05) were observed. No other appreciable differences were found. At the study onset, negative correlations were found between L-BMD Z-score and serum ALP, BAP, and phosphorus and between TB-BMD Z-score and urine uCa/Cr. After 1 year, L-BMD Z-score correlated negatively with serum BAP and OC, and positively with serum 25(OH)D3. Multivariate analysis showed that BAP was the strongest predictor of L-BMD Z-score (beta=-0.49; p<0.05). We conclude that children with nephrotic syndrome treated with corticosteroids are at risk of bone mass loss. Serum BAP concentration seems to be a good indicator of spongy bone metabolism in these children, who should be supplemented with vitamin D in an adjustable dose, possibly higher than 800 IU/24 h to prevent osteopenia.

The role of natural killer cells in pathogenesis of autoimmune diseases.

There is growing evidence that NK cell-mediated immunoregulation plays an important role in the control of autoimmunity. NK cells are a subset of lymphocytes that generally contribute to innate immunity but have also a great impact on the function of T and B lymphocytes. The major role of NK cells is cytotoxic reaction against neoplastic, infected and autoreactive cells, but they regulatory function seems to play more important role in the pathogenesis of autoimmune diseases. Numerous studies suggested the involvement of NK cells in pathogenesis of such a common autoimmune diseases as juvenile rheumatoid arthritis, type I diabetes and autoimmune thyroid diseases. The defects of NK cells regulatory function as well as cytotoxic abilities are common in patients with autoimmune diseases with serious consequences including HLH hemophagocytic lymphocytosis (HLH) and macrophage activation syndrome (MAS). The early diagnosis of NK cells defect responsible for the loss of the protective abilities is crucial for the prevention of life-threatening complications and implementation of necessary treatment.

Recurrent respiratory tract infections in children - analysis of immunological examinations.

BACKGROUND: Paediatric respiratory tract infections are among the most common reasons for preschool and school absences and visits to physicians. The disease mainly involves the upper respiratory tract and is associated with fever, cough, sore throat, and running nose. Children with recurrent respiratory infections (RRI), which are defined as more than six serious diseases a year, are a difficult diagnostic challenge. The aim of this study was to assess immunological deviations in laboratory tests performed in children with RRI. MATERIAL AND METHODS: In the retrospective study 25 children suffering from recurrent respiratory tract infection, aged 4.1 ±2.3 years, 13 boys and 12 girls, were involved. For all children chemiluminescence of granulocytes and immunophenotyping of lymphocytes from peripheral blood were examined. An immunophenotype of peripheral blood lymphocytes involved evaluation of T cell, B cells, and NK cells, examined with flow cytometry. RESULTS: Eleven of the studied children had decreased chemiluminescent response to stimulants, normal response was found for nine children, and five children had an increased result of the test. Five of the 25 children had decreased B cells number, and five had decreased number of T cells including decrease of CD4, as well as CD8 positive cells. Children with decreased chemiluminescence had more frequent neutropaenia than children with normal or increased chemiluminescent response, p < 0.05 (exact Fisher test). CONCLUSIONS: Recurrent respiratory tract infection could be associated with improper neutrophils response to pathogens, and immunological examination should be performed to find the reason for the increased number of infections in a year.

Serum fibroblast growth factor 23 and calcium-phosphorus metabolism parameters in children with chronic kidney disease - preliminary report.

INTRODUCTION: In chronic kidney disease (CKD) the function of all factors regulating mineral metabolism is disturbed, leading inevitably to renal osteodystrophy and vascular calcification. The aimof the study is to assess concentrations of fibroblast growth factor 23 (FGF 23), osteoprotegerin (OPG) and other parameters of calcium-phosphate metabolism in children with CKD. MATERIAL AND METHODS: 37 children with CKD 3-5, aged 1.6-17 years were included in the study. In all children serum levels of calcium (sCa), phosphate (sP), creatinine, alkaline phosphatase (ALP), FGF 23, intact parathormone (PTH), OPG and receptor activator nuclear factor κB ligand (RANKL) were measured. RESULTS: Total calcium concentration was within normal limits in all children included in this study. Hyperphosphatemia was found in 2 children from group CKD 3 (12%), 6 from CKD 4 (54%) and 1 from CKD 5 (11%). FGF 23 level increased consecutively in subsequent CKD stages achieving the highest values in CKD 5 group. In all children with CKD, serum levels of OPG were correlated with FGF 23. In children with CKD 3-4 negative correlation between FGF 23 and PTH (r=-0.45; p=0.02) and positive correlation between FGF 23 and RANKL (r=0,59; p=0.006) has been found. Positive correlation between OPG concentration and HCO3 -and BE levels has been observed, as well as negative correlation between RANKL/OPG ratio and HCO3 -and BE levels. CONCLUSION: Despite maintaining serum calcium, phosphorus and PTH levels within recommended limits, elevated levels of FGF 23 and OPG were observed in children with chronic kidney disease, especially in it's end-stage.

Six-minute walk test reflects neurohormonal activation and right ventricular function in systemic sclerosis patients.

OBJECTIVES: Heart and pulmonary involvement is a leading cause of systemic sclerosis (SSc)-related deaths. The six-minute walk test (6MWT) is a simple and reproducible test commonly used to evaluate exercise capacity. We tried to assess a potential relationship between exercise capacity assessed by 6MWT, echocardiographic parameters of right ventricular function and serum levels of endothelin-1 and NTproBNP. METHODS: We prospectively studied 111 consecutive patients (101F, 10M, age 54.2±13.8 years) with diagnosed SSc (mean disease duration 9±12.4 years) and a group of 21 age-matched subjects (18F, 3M, age 49.3±10.5 years). In addition to routine evaluation, 6MWT and transthoracic echocardiography (Phillips iE 33) were performed. We also measured serum endothelin-1 (Human Endothelin-1 immunoassay R & D Systems) and NT-proBNP levels (Elecsys pro-BNP immunoassay; Roche Diagnostics). RESULTS: The mean 6MWT distance was significantly shorter in the SSc group than in the controls (562.8±60.3 vs. 514.7±102.5 m, p=0.03). In the SSc group 6MWT distance correlated with ET-1 (r=-0.5, p<0.0001), NTproBNP (r=-0.4, p=0.0008) levels, and echocardiographic indices AcT (r=0.4, p=0.0002) and TRPG (r=-0.4, p=0.0011). Moreover, in patients with 6MWT distance <450 m NTproBNP and endotothelin-1 levels were significantly higher than in patients with distance >450 m (311.2, 31.1-17237 vs. 105.3, 5-17670 pg/ml, p=0.0138 and 2.9±2.2 vs. 1.4±0.7 pg/ml, p=0.0032). CONCLUSIONS: Decreased exercise capacity significantly correlates with biochemical and echocardiographic parameters of right ventricular dysfunction and neurohormonal activation providing a potential link for neuroendocrine derangement in patients with SSc.

Delay in the measurement of eosin-5'-maleimide (EMA) binding does not affect the test result for the diagnosis of hereditary spherocytosis.

BACKGROUND: The eosin-5'-maleimide (EMA) binding test is a flow cytometric test widely used to detect hereditary spherocytosis (HS). EMA binds to plasma membrane proteins of red blood cells (RBCs), mainly to band 3 protein. The mean fluorescence of EMA-stained RBCs in HS patients is lower when compared with control RBCs due to the decreased amount of target proteins. EMA dye in aqueous solution is sensitive to light and high temperature. Its fluorescence can decrease when exposed to light or ambient temperatures higher than 4°C. The aim of the study was to evaluate the stability of fluorescence readings of EMA-labeled RBCs over a period of 24 h. METHODS: The EMA test was performed in peripheral blood from 35 patients with microcytic anemia (five with HS, and 30 without HS). Peripheral blood samples were stained immediately after blood collection and analyzed using a flow cytometer at three time points: 0, after 1 and 24 h of storage at 4°C in the darkness. The results are presented as the percentage of normal control RBCs fluorescence. Flow cytometric studies were performed with Cytomics FC500 (Beckman Coulter, USA). RESULTS: In HS patients the mean result of the test reached 66.72%±9.26% of normal controls, and in non-HS patients the EMA result was 99.48%±5.03% of normal control cells. The results of patients with HS were 66.72%±9.26%, 66.90%±10.24% and 67.86%±11.31% at 0 h, and after 1 and 24 h of storage, respectively. The results obtained from non-HS patients at time 0, after 1 and 24 h of storage reached 99.48%±5.03%, 99.49%±5.34% and 99.78%±6.13%, respectively. There was no difference between the results from each time point in samples from patients with or without HS. CONCLUSIONS: Results of the EMA binding test do not depend on storage time of stained samples when stored at 4°C up to 24 h after staining.