Very early and early nutrition in children with pancreatitis-A randomised trial.

OBJECTIVES: The aim of our study was to assess the impact of the very early introduction of refeeding on the course of acute pancreatitis (AP) in children. Additionally, we evaluated the effect of nutrition on inflammatory markers, including cytokines. METHODS: This prospective randomised study was conducted in three university hospitals in Poland. Patients, aged 1-18 years with AP, were randomised into two groups: A-refeeding within 24 h of hospital admission (very early), and B-refeeding at least 24 h after admission (early nutrition). The severity of AP was assessed after 48 h. The serum concentrations of four cytokines (tumour necrosis factor α [TNFα], interleukin-1ß [IL-1ß], interleukin-6 [IL-6] and interleukin-8 [IL-8]) and C-reactive protein, as well as the activity of amylase, lipase and aminotransferases, were measured during the first 3 days of hospitalisation. RESULTS: A total of 94 children were recruited to participate in the study. The statistical analysis included 75 patients with mild pancreatitis: 42-group A and 33-group B. The two groups did not differ in the length of hospitalisation (p = 0.22), AP symptoms or results of laboratory tests. Analysis of cytokine levels was conducted for 64 children: 38-group A and 26-group B. We did not find a difference in concentrations of the measured cytokines, except for IL-1ß on the third day of hospitalisation (p = 0.01). CONCLUSIONS: The time of initiation of oral nutrition within 24 h (very early) or after 24 h (early) from the beginning of hospitalisation had no impact on the length of hospitalisation, concentrations of TNF-α, IL-1ß, IL-6 and IL-8, activity of amylase and lipase or occurrence of symptoms in children with mild AP.

Peripheral blood lymphocyte immunophenotyping (TBNK) - a comparison of BD FACSCanto II and BD FACSLyric flow cytometry analysers.

Introduction: Flow cytometry immunophenotyping is a common laboratory technique for evaluating lymphocyte subpopulations. Its result remains an important diagnostic tool in various medical fields. Cytometric tests are performed in many laboratories, making the comparability between different devices using the same method an important aspect. We aimed to compare the results of lymphocyte immunophenotyping (lymphocytes B, T, Th and Tc, NK cells) between two different flow cytometers. Material and methods: The study included 93 patients of the Children's Teaching Hospital of the Medical University of Warsaw and 9 Multi-Check control results. The method of lymphocyte subpopulation assessment was based on fluorescent flow cytometry immunophenotyping, using a BD Multitest 6-color TBNK kit (Becton Dickinson). We compared BD FACSCanto II and BD FACSLyric analysers (Becton Dickinson). For data analysis, we used Spearman's rank correlation, Bland-Altman plot and Passing-Bablok regression. Results: Spearman's rank correlation showed a strong interrelation for all analysed parameters (0.808-0.985). In the Passing-Bablok regression analysis, all examined parameters showed linear dependence with the slope values close to 1 (0.940-1.134). Bland-Altman coefficient values were within the range of 2.94-8.62% with half of them being above 5% (T, Tc, Th, B, NKT absolute values and B percentage values). Conclusions: The results from both cytometers can be considered equivalent, but it should be noted that one of the statistical methods showed some deviations, presumably primarily due to the evaluators' different gating techniques. The training of specialists performing these tests requires more attention.

Hyperthermia Enhances Adeno-Associated Virus Vector Transduction Efficiency in Melanoma Cells.

Gene therapy perfectly fits in the current needs of medicine for patients with melanoma. One of the major challenges of gene therapy is to increase gene transfer. The role of hyperthermia in the improvement of AAV (adeno-associated virus) transduction efficiency has been indicated. The aim of the present study was to assess the transduction efficacy of melanoma cell lines (A375, G-361, and SK-MEL-1) with the use of the rAAV/DJ mosaic vector under hyperthermia conditions. The analysis of changes in the transduction efficacy and expression of HSPs (heat shock proteins) and receptors for AAV was performed. The transduction was performed at 37 °C and at 43 °C (1 h). Hyperthermia enhanced gene transfer in all the tested cell lines. The most efficient transducing cell line under hyperthermia was A375 (increase by 17%). G361 and SK-MEL-1 cells showed an increase of 7%. The changes in the expression of the AAV receptors and HSPs after hyperthermia were observed. A key role in the improvement of gene transfer may be played by AAVR, HSPB1, HSP6, DNAJC4, HSPD1, HSPA8, HSPA9, HSP90AB1, and AHSA1. This study showed the possibility of the use of hyperthermia as a factor enabling the stimulation of cell transduction with rAAV vectors, thereby providing tools for the improvement in the efficacy of gene therapy based on rAAV.

Influence of blood sample storage and different types of anticoagulants on results of NK cytotoxicity tests.

Introduction: Natural killer (NK) cells are important players in the human immune response. Impaired NK function may lead to serious, life-threatening conditions. Defects may be consequences of genetic mutations or results of secondary factors such as infections, malignancies and autoimmune diseases. The cytotoxicity test is very useful, but its accessibility is limited to special immunological laboratories. Blood samples are often transported to remote centers, which takes time and requires special conditions.The aim of this study was to compare cytotoxicity assay results between samples preserved with three different anticoagulants to standardize the diagnostic procedure. Material and methods: Peripheral blood from healthy donors was taken with three anticoagulants: heparin, K2EDTA and citrate. Peripheral blood mononuclear cells (PBMC) were isolated and tested directly after blood drawing and after 24-hour storage. Cytotoxic abilities of NK cells were tested in 4 h co-culture with K562. NK cytotoxicity was measured by flow cytometry. Results: In most cases of analyzed healthy donors, cytotoxicity results were similar regardless of type of anticoagulant. However, the highest mean values were obtained in samples with citrate. There was a significant decrease in cytotoxicity after 24 hours of storage of the whole blood at ambient temperature. The mean drop in cytotoxicity results was substantial for all anticoagulants: 76% for heparin, 67% for citrate and 70% for EDTA. Conclusions: Results of spontaneous NK cytotoxicity seem to be affected by the anticoagulants used for blood protection. Commercial instant cytotoxicity testing and delayed analysis after blood storage gave the highest results in blood with sodium citrate.

Recurrent assessment of lymphocyte subsets in 32 patients with multisystem inflammatory syndrome in children (MIS-C).

BACKGROUND: Lymphopenia is a hallmark of multisystem inflammatory syndrome in children (MIS-C). We aimed to characterize lymphocyte subsets' shifts and their correlations with other severity markers of MIS-C. METHODS: In this prospective cross-sectional study, we performed peripheral lymphocyte phenotyping in 32 patients with MIS-C. We analyzed lymphocyte subsets at three time points of the disease: the acute (A), convalescent (B), and recovery (C) phases. Based on age-normalized lymphocyte counts, we distinguished two groups of patients: "the mild" (higher lymphocyte counts) and "the severe" (lower lymphocyte counts). In addition, we examined differences between these groups regarding other severity markers. RESULTS: In phase A, 84% of children had lymphopenia. Decreased absolute counts of CD3, CD4, and CD8 cells were observed in, respectively, 88%, 72%, and 84% of patients. The natural killer cells were decreased in 63% and CD19 in 59% of children. "The severe" group had significantly higher procalcitonin and troponin I levels and lower platelets and albumin. Moreover, "the severe" group had hypotension more frequently (73% vs. 20%, p = .008). In phase B, all lymphocyte counts increased, and 32% of children had lymphocytosis. The increase of CD3, CD4, and CD8 counts correlated with some laboratory severity markers (hemoglobin, procalcitonin, D-dimer, lactate dehydrogenase, N-terminal prohormone of brain natriuretic peptide, albumin), but not with steroid use. In phase C, most children had normal lymphocyte counts. CONCLUSIONS: Substantial shifts in lymphocyte counts during MIS-C apply most to T lymphocytes and correlate with the disease severity markers, particularly hypotension prevalence. A proportion of children with MIS-C develops transient lymphocytosis during convalescence.

Circulating calcification inhibitors are associated with arterial damage in pediatric patients with primary hypertension.

BACKGROUND: Circulating calcification inhibitors: fetuin A (FA) and osteoprotegerin (OPG) together with soluble ligand of receptor activator of nuclear factor kappa-B (sRANKL) have been linked to vascular calcifications and arterial damage. This study aimed to evaluate relationships between FA, OPG, sRANKL, and arterial damage in children with primary hypertension (PH). METHODS: In this cross-sectional single-center study, calcification inhibitors (FA, OPG, sRANKL) levels were measured in blood samples of 60 children with PH (median age 15.8, IQR: [14.5-16.8] years) and 20 age-matched healthy volunteers. In each participant, peripheral and central blood pressure evaluation (BP) and ambulatory BP monitoring (ABPM) were performed. Arterial damage was measured using common carotid artery intima media thickness (cIMT), pulse wave velocity (PWV), augmentation index (AIx75HR), and local arterial stiffness (ECHO-tracking-ET) analysis. RESULTS: Children with PH had significantly higher peripheral and central BP, BP in ABPM, thicker cIMT, higher PWV, and AIx75HR. FA was significantly lower in patients with PH compared to healthy peers without differences in OPG, sRANKL, and OPG/sRANKL and OPG/FA ratios. In children with PH, FA level correlated negatively with cIMT Z-score and ET AIx; sRANKL level correlated negatively with ABPM systolic blood pressure (SBP), SBP load, diastolic BP load, and AIx75HR; OPG/sRANKL ratio correlated positively with SBP load, while OPG/FA ratio correlated positively with ET AIx. In multivariate analysis, FA was a significant determinant of cIMT (mm) and cIMT Z-score. CONCLUSIONS: This study reveals that in children with primary hypertension, arterial damage is related to lower fetuin A concentrations.

Plerixafor combined with G-CSF for stem cell mobilization in children qualified for autologous transplantation- single center experience.

Failure of autologous peripheral blood CD34+ stem cells collection can adversely affect the treatment modality for patients with hematological and nonhematological malignant diseases where high dose chemotherapy followed by hematopoietic stem cell transplantation has become part of their treatment. Plerixafor in conjunction with G-CSF is approved for clinical use as a mobilization agent. The clinical efficacy of Plerixafor in CD34+ cells collection was analyzed in our institution. A total of 13 patients aged 1-15,5 years received Plerixafor in combination with G-CSF: 7 with neuroblastoma, 2 with Ewing's sarcoma and single patients with Hodgkin's lymphoma, germ cell tumor, retinoblastoma and Wilms tumor. Twelve patients (923%) achieved CD34+ cell counts of ≥ 20 × 106/L after 1-7 doses of Plerixafor. The average 9,9 - fold increase in number of CD34+ cells were achieved following the first dose and 429 - fold after second dose of plerixafor. Among the 13 patients, 12 yielded the minimum required cell collection of 2 × 106/kg within an average of 2 doses of Plerixafor. The mean number of apheresis days was 1.75. The median total number of collected CD34+ cells was 982 × 106/kg. Plerixafor enables rapid and effective mobilization, and collection of sufficient number of CD34+ cells.

Extracellular Vesicles in Allergic Rhinitis and Asthma and Laboratory Possibilities for Their Assessment.

Currently, extracellular vesicles (EVs) have been implicated in the etiopathogenesis of many diseases, including lung disorders, with the possibility of diagnostic and therapeutic applications. The analysis of EV in respiratory tract diseases faces many obstacles, including material collection from airways, standardization of isolation techniques, detection methods, the analysis of their content, etc. This review focuses on the role of extracellular vesicles in the pathogenesis of atopic respiratory diseases, especially asthma, with a special focus on their clinical applicability as a diagnostic tool. We also summarize available laboratory techniques that enable the detection of EVs in various biological materials, with particular emphasis on flow cytometry. The opportunities and limitations of detecting EV in bronchoalveolar lavage fluid (BALF) were also described.

Relationship between Gd-IgA1 and TNFR1 in IgA nephropathy and IgA vasculitis nephritis in children - multicenter study.

AIM OF THE STUDY: To evaluate the relationship between serum Gd-IgA1 (sGd-IgA1) and serum and urine TNFR1 (sTNFR1, uTNFR1) levels as possible prognostic factors in IgA nephropathy (IgAN) and IgA vasculitis nephritis (IgAVN). MATERIAL AND METHODS: From 299 patients from the Polish Registry of Pediatric IgAN and IgAVN, 60 children (24 IgAN and 36 IgAVN) were included in the study. The control group consisted of 20 healthy children. Proteinuria, haematuria, serum creatinine as well as IgA and C3 levels were measured and glomerular filtration rate (GFR) was calculated at onset and at the end of the follow-up. Kidney biopsy findings were evaluated using the Oxford classification. Serum Gd-IgA1 and serum and urine TNFR1 levels were measured at the end of follow-up. RESULTS: Serum Gd-IgA1 level was significantly higher in IgAN and IgAVN patients in comparison to the control group. Urine TNFR1 was significantly higher in IgAN than in IgAVN and the control group. We did not observe any differences in sTNFR1 level between IgAN, IgAVN and control groups. We found a positive correlation between Gd-IgA1 and creatinine (r = 0.34), and negative between Gd-IgA1 and GFR (r = -0.35) at the end of follow-up. We observed a negative correlation between uTNFR1/creatinine log and albumin level and protein/creatinine ratio. We did not find any correlations between Gd-IgA1 and TNFR1. CONCLUSIONS: The prognostic value of sGd-IgA1 in children with IgAN and IgAVN has been confirmed. TNFR1 is not associated with Gd-IgA1 and is not a useful prognostic marker in children with IgAN/IgAVN and normal kidney function.

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.

Expression of programmed cell death protein 1 and T-cell immunoglobulin- and mucin-domain-containing molecule-3 on peripheral blood CD4+CD8+ double positive T cells in patients with chronic hepatitis C virus infection and in subjects who spontaneously cleared the virus.

Chronic hepatitis C virus (HCV) infection is characterized by increased proportion of CD4+CD8+ double positive (DP) T cells, but their role in this infection is unclear. In chronic hepatitis C, immune responses to HCV become functionally exhausted, which manifests itself by increased expression of programmed cell death protein 1 (PD-1) and T-cell immunoglobulin- and mucin-domain-containing molecule-3 (Tim-3) on T cells. The aim of our study was to determine PD-1 and Tim-3 phenotype of DP T cells in subjects with naturally resolved and chronic HCV infection. Peripheral blood mononuclear cells from 16 patients with chronic infection and 14 subjects who cleared HCV in the past were stained with anti-CD3, anti-CD4, anti-CD8, anti-PD-1 and anti-Tim-3 antibodies and, in 12 HLA-A*02-positive subjects, MHC class I pentamer with HCV NS31406 epitope. In chronic and past HCV infection, proportions of total DP T cells and PD-1+ DP T cells were similar but significantly higher than in healthy controls. DP T cells were more likely to be PD-1+ than either CD4+ or CD8+ single positive (SP) T cells. HCV-specific cells were present in higher proportions among DP T cells than among CD8+ SP T cells in both patient groups. Furthermore, while the majority of HCV-specific DP T cells were PD-1+, the proportion of HCV-specific CD8+ T cells which were PD-1+ was 4.9 and 1.9 times lower (chronic and past infection, respectively). PD-1 and Tim-3 were predominantly expressed on CD4high CD8low and CD4low CD8high cells, respectively, and co-expression of both markers was uncommon.

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.

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.

The influence of agents differentiating HL-60 cells toward granulocyte-like cells on their ability to release neutrophil extracellular traps.

Studies on neutrophil extracellular traps (NETs) are challenging as neutrophils live shortly and easily become activated. Thus, availability of a cell line model closely resembling the functions of peripheral blood neutrophils would be advantageous. Our purpose was to find a compound that most effectively differentiates human promyelocytic leukemia (HL-60) cells toward granulocyte-like cells able to release NETs. HL-60 cells were differentiated with all-trans retinoic acid (ATRA), dimethyl sulfoxide (DMSO) or dimethylformamide (DMF) and stimulated with phorbol 12-myristate 13-acetate (PMA) or calcium ionophore A23187 (CI). Cell differentiation, phagocytosis and calcium influx were analyzed by flow cytometry. Reactive oxygen species production and NETs release were measured fluorometrically and analyzed microscopically. LC3-II accumulation and histone 3 citrullination were analyzed by western blot. ATRA most effectively differentiated HL-60 cells toward granulocyte-like cells. ATRA-dHL-60 cells released NETs only upon PMA stimulation, DMSO-dHL-60 cells only post CI stimulation, while DMF-dHL-60 cells formed NETs in response to both stimuli. Oxidative burst was induced in ATRA-, DMSO- and DMF-dHL-60 cells post PMA stimulation and only in DMF-dHL-60 cells post CI stimulation. Increased histone 3 citrullination was observed in stimulated DMSO- and DMF-, but not in ATRA-dHL-60 cells. The calcium influx was diminished in ATRA-dHL-60 cells. Significant increase in autophagosomes formation was observed only in PMA-stimulated DMF-dHL-60 cells. Phagocytic index was higher in ATRA-dHL-60 cells than in control, DMSO- and DMF-dHL-60 cells. We conclude that ATRA, DMSO and DMF differentiate HL-60 in different mechanisms. DMF is the best stimulus for HL-60 cell differentiation for NETs studies.

Association Between Vitamin D and Carboxy-Terminal Cross-Linked Telopeptide of Type I Collagen in Children During Growth Hormone Replacement Therapy.

Growth hormone and insulin-like growth factor-1 (IGF-1) play a crucial role in the regulation of bone turnover. Adequate vitamin D status supports proper bone remodeling, leading to normal longitudinal bone growth and normal peak bone mass. The aim of this study was to evaluate the association between serum 25-hydroxyvitamin D [25(OH)D] and carboxy-terminal cross-linked telopeptide of type I collagen (ICTP) in children and adolescents with growth hormone deficiency at baseline and during recombinant human growth hormone (rhGH) replacement therapy. The study was prospective and included 30 children and adolescents aged 5 to 17 years. Concentrations of 25(OH)D, ICTP, and IGF-1 were measured at baseline and during the first year of rhGH therapy. Baseline serum 25(OH)D concentration correlated with ICTP concentrations during the first trimester of rhGH therapy (r = 0.38, p < 0.050); the correlation was stronger in the second trimester of therapy (r = 0.6, p = 0.002). We conclude that proper vitamin D status is important in reaching the adequate dynamics of bone remodeling during growth, which is essential to achieve a catch-up growth during rhGH therapy.

Osteoprotegerin, Receptor Activator of Nuclear Factor Kappa B Ligand, and Growth Hormone/Insulin-Like Growth Factor-1 Axis in Children with Growth Hormone Deficiency.

The growth hormone/insulin-like growth factor-1 (GH/IGF-1) axis is involved in the regulation of the receptor activator of nuclear factor kappa B ligand (RANKL)/RANK/osteoprotegerin (OPG) system, but the exact mechanism of the associations is not fully explained. In this study we investigated the serum OPG and total sRANKL concentrations in short children who had differences in GH secretory status. We also investigated the associations between the GH/IGF-1 and OPG/RANKL systems in GH-deficient children during GH treatment. There were no significant differences in any anthropometric or biochemical parameters evaluated between the GH-deficient and GH-sufficient children. The OPG content and total alkaline phosphatase (ALP) activity increased significantly after the initiation of GH treatment, while total sRANKL remained unchanged. The variables baseline BMI SDS for height-age (ß = 0.42; p < 0.05), baseline ALP activity (ß = 0.36; p < 0.05), weight SDS for height-age at 6 months of GH treatment (ß = 1.86; p < 0.01), and total ALP activity at 6 months of GH treatment (ß = 0.48, p < 0.01) were identified as independent predictors of ΔOPG6-month-baseline. We conclude that OPG and total sRANKL concentrations are independent from GH secretory status in short children. OPG elevation during GH treatment is independently associated with total ALP activity and nutritional status in GH-deficient children.

[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.

Utility of leucocyte antigens in distinguishing between bacterial and viral infection in children.

INTRODUCTION: Accurate diagnosis of bacterial and viral infection is very difficult. Unfortunately, there is still no quick and discriminative diagnostic test that would help clinicians in establishing the diagnosis and taking a decision on treatment. The aim of the study was to compare the expression of antigens on phagocytes, which are involved in the first defence line during bacterial and viral infections in children, as a potential tool to distinguish the etiology of the infection. MATERIAL AND METHODS: The expression of CD35, CD32, CD88, and MHC class I on phagocytes in 49 blood samples from children with high fever and suspected infection as well as 19 healthy children (control group) was assessed by flow cytometry. Thirty-three children were diagnosed with bacterial and 16 with viral infection. Expression of antigens was analysed on a FACSCanto II flow cytometer according to mean fluorescence intensity (MFI) and antibody binding cites (ABC). RESULTS: Significant differences were observed for the following: CD32, CD35, CD88, and MHCI on granulocytes; CD32, CD35, CD88 on monocytes; and MHC-I ratio between groups were observed. The obtained results did not allow us to establish valuable score points for distinguishing between bacterial and viral infections. Classification and a regression tree using CD88 expression on granulocytes and CRP was developed. It enabled us to differentiate between the origin of infection with sensitivity and specificity of more than 90%. CONCLUSIONS: Utility of use of wide range antigens' expression on phagocytes for distinguishing between bacterial and viral infection in children has limited value. More adequate seems to be use of CD88 expression on granulocytes linked with CRP value.