Monitoring of treatment with L-asparaginase in children with acute lymphoblastic leukaemia, with a focus on silent inactivation and its influence on the treatment outcome.

Introduction: The aim of the study was to analyse the frequency of silent inactivation and allergic reaction to asparaginase (ASP) and its impact on treatment results in patients with lymphoblastic leukaemia. Material and methods: Seventy patients with acute lymphoblastic leukaemia treated with ASP were enrolled in the study. Asparaginase activity was monitored. The patients were switched to another ASP formulation after allergy or inactivation. The treatment results were analysed. Results: Silent inactivation of native E. coli ASP was diagnosed in 5 patients (7%) and allergy in 34 patients (49%), and these patients were switched to pegylated ASP (PEG-ASP). Silent inactivation of PEG-ASP occurred in 8 patients (23%) and allergy in 6 patients (17%). Eight children continued therapy with Erwinase, and 4 did not switch to Erwinase after inactivation of PEG-ASP. Allergy to Erwinase occurred in 2 patients (22%); there was no inactivation. No significant differences in outcome were found between the groups of patients with and without allergy or silent inactivation of ASP. Due to regular monitoring and switching to other ASP preparations after allergy or silent inactivation, therapeutic activity was ensured in almost all patients. Conclusions: Monitoring of ASP activity is crucial to recognize silent inactivation and to guarantee treatment effectiveness by switching to other ASP preparations.

Intravenous, Perioperatively Administered Lidocaine Regulates Serum Pain Modulators' Concentrations in Children Undergoing Spinal Surgery.

OBJECTIVES: We analyzed the influence of perioperative, intravenous (i.v.) lidocaine infusion as a part of multimodal anesthesia on concentrations of selected pain modulators. DESIGN: An observational study. SETTING: University Children's Hospital in Cracow, Poland, from May 2015 to May 2018. SUBJECTS: Forty-four children undergoing extensive spinal surgery, divided into two groups after surgery: the study group (N = 23), anesthetized generally with lidocaine as a co-analgesic, and the control group (N = 22), anesthetized generally without lidocaine. METHODS: We assessed proinflammatory mediators like neuron growth factor (NGF), high mobility group box 1 (HMGB1), interleukin 6 (IL-6), and FOS protein before, immediately after, six hours and 12-15 hours after surgery. We evaluated pain intensity at corresponding time points using a 10-point numerical/graphical scale. RESULTS: We observed that children in the lidocaine group had reduced pain intensity in the resting state and during movement until six hours after surgery when compared with controls. We found lower NGF concentrations in the lidocaine group vs controls only at six hours after surgery. Mean HMGB1 concentrations during the postoperative period in the study group were relatively stable, whereas we observed significant increases at six hours after surgery and a slight decrease at 12-15 hours after surgery in the control group. IL-6 concentrations at six hours were lower in lidocaine patients when compared with controls. We noted a negative correlation between HMGB1, NGF, Il-6, and lidocaine concentrations after surgery. We did not find any differences in FOS protein concentrations between the groups. CONCLUSIONS: Our findings suggest that intraoperative and postoperative i.v. lidocaine administration as a part of multimodal anesthesia may reduce inflammatory-dependent postoperative pain intensity.

Composition of tissue fatty acids of phospholipids depends on tumor localization and disease progression in colorectal patients.

OBJECTIVES: Fatty acids play a role in development and progression of colon cancer. The aim of this study was to assess the relation between tissue fatty acids (saturated fatty acids, unsaturated fatty acids, the ratio of C18 to C18:1 - index of fatty acids saturation, SI), colorectal tumor localization and disease progression. METHODS AND RESULTS: Total of 49 patients (14 with proximal colon, 13 with distal colon and 22 with rectal tumor localization) were studied. One year after surgery 24 patients had the disease progression. Tissue levels of saturated fatty acids and unsaturated fatty acids were measured before surgery by gas-chromatography. These fatty acids were determined in cancerous tissue (CA) and non-cancerous tissue (NCA). The most significant differences in the mean values of fatty acids of phospholipids between CA and NCA in patients with proximal tumor localization were noted. The mean value of C18 was significantly lower while C18:1 was significantly higher in CA as compared to NCA in patients without disease progression (p<0.02; p<0.03; respectively). SI was significantly lower in CA as compared to NCA only in patients without disease progression (p<0.02). CONCLUSION: Fatty acids of tissue phospholipids' fraction, as well as SI, strongly depend on tumor localization and might be useful as potential markers of the disease progression in colorectal cancer patients.

Plasma Free Fatty Acids and their Binding Proteins in Preterm Infants.

BACKGROUND/AIMS: The objective of the study was to evaluate the circulating concentrations of plasma free fatty acids (FFA), fatty acid binding proteins: FABP-1 and FABP-4 in preterm infants depending on different feeding protocol. METHODS: A total of 43 premature infants (≤34 weeks) were enrolled in the study, and divided into 3 subgroups: nursed while staying in the department (53%), breast-fed only during the first 24 h (16%), and formulafed from the beginning (31%). The control group consisted of 12 healthy, full-term, breast-fed newborns. Blood samples were collected after delivery and 1 month later. We measured plasma concentrations of FFA, FABP-1, and FABP-4. RESULTS: FFA plasma concentrations were significantly lower in preterm babies when compared to control group (p = 0.003) in the prenatal period. After 1 month, a significant decrease in FFA concentration was noted in all groups of preterm babies independently from feeding protocol. After a month, breast-fed preterm infants and controls had significantly lower FABP-1 levels than preterm formula-fed infants (all p < 0.05), while the highest concentrations of FABP-4 were noted in formula-fed preterm infants when compared to breast-fed preterm infants and the control group (all p < 0.05). CONCLUSIONS: Prematurity is connected with disturbances in plasma FFA concentrations. FABP-1, as well as FABP-4, plasma levels in preterm infants depend on feeding protocol.

Shall we diagnose metabolic syndrome in adolescents?

BACKGROUND: The clinical value of the diagnosis of metabolic syndrome (MS) in children and adolescents remains unclear. The aim of the present study was to assess the occurrence of metabolic complications, other than included in 2007 IDF MS definition, in obese children and adolescents METHODS: The study included 75 (33 boys) obese adolescents (mean age 13.9 years, mean BMI SDS 4.49). Classical (fasting glucose, TGL, HDL, blood pressure) and non classical (insulin resistance [HOMA-IR], creatinine, AST, ALT, uric acid, fibrinogen, liver US and 24h BP profile) risk factors were compared between groups with and without MS. 15(8 boys) met the 2007 IDF criteria for MS. RESULTS: Patients with MS presented with significantly lower: BMI SDS (4.2 vs. 5.8, p=0.02), mean 24h SBP (0.8 vs. 1.0, p=0.03), and uric acid level (352.1 vs. 414.0, p=0.01). In both groups a significant percentage of abnormal results of 24hABPM (up to 42.9 and 57.6%), insulin resistance (85.7 % and 61.1%), non alcoholic fatty liver disease (57.4 % and 38.9 %) and hyperuricemia (69.2 % and 55.3%) was observed. CONCLUSION: Recognizing the metabolic syndrome in adolescents does not provide any additional clinical benefits. It seems that in every obese child a wide, personalized diagnostic work-up should be performed.

[B-type natriuretic peptide as a marker of cardiac dysfunction in children with chronic kidney disease]. / Peptyd natriuretyczny typu B jako marker uszkodzenia serca u dzieci z przewlekla choroba nerek.

Left ventricular hypertrophy is the most common organ damage in children with chronic kidney disease (CKD). AIM: The aim of the study was to assess the usefulness of B-type natriuretic peptide (BNP) as a marker of heart injury in children with CKD. MATERIALS AND METHODS: We included 66 children (41 boys and 25 girls) aged 0.7 to 18.6 (median 11.6) years with CKD stage 1-5. The concentrations of urea, creatinine, cystatin C and BNP in blood serum were assessed, and the estimated glomerular filtration rate (eGFR) was calculated from the Schwartz and Filler formulas. Patients were divided into groups depending on the CKD stage [group 1: CKD stages 1 + 2 (GFR> 60 ml/min/1.73 m2), group 2: stage 3 (GFR = 30-59 ml/min/1.73 m2), group 3: CKD stage 4 (GFR 15-29 ml/min/ 1.73 m2), group 4 - stage 5 (dialyzed children)]. On the basis of echocardiography, the left ventricular mass (LVM) was calculated, which was indexed for height (left ventricular mass index, LVMI). Left ventricular hypertrophy (LVH) was diagnosed if the LVMI value was > 95th percentile for sex and age. RESULTS: Depending on the CKD stage the median BNP concentrations for group 1, group 2, group 3, and group 4 were 2.5 pg/ml, 6.0 pg/ml, 9.3 pg/ml and 18.0 pg/ml, and the LVH prevalence 27.3%, 33.3%, 60.0% and 63.6% , respectively. Significant correlations between BNP concentration and LVH expressed by LVMI (R=0.256, p=0.038), creatinine (R=0.453, p<0.001), cystatin (R=0.494, p<0.001) and eGFR (R=-0.473, p<0.001) were found. CONCLUSIONS: In children with chronic kidney disease, BNP is an indicator of heart failure correlating with renal function parameters and left ventricular mass index.

Simplification of intradermal skin testing in Hymenoptera venom allergic children.

BACKGROUND: The direct comparison between children and adults with Hymenoptera venom anaphylaxis (HVA) has never been extensively reported. Severe HVA with IgE-documented mechanism is the recommendation for venom immunotherapy, regardless of age. OBJECTIVE: To determine the differences in the basic diagnostic profile between children and adults with severe HVA and its practical implications. METHODS: We reviewed the medical records of 91 children and 121 adults. RESULTS: Bee venom allergy was exposure dependent, regardless of age (P < .001). Atopy was more common in children (P = .01), whereas cardiovascular comorbidities were present almost exclusively in adults (P = .001). In the bee venom allergic group, specific IgE levels were significantly higher in children (29.5 kUA/L; interquartile range, 11.30-66.30 kUA/L) compared with adults (5.10 kUA/L; interquartile range, 2.03-8.30 kUA/L) (P < .001). Specific IgE levels for culprit insect venom were higher in bee venom allergic children compared with the wasp venom allergic children (P < .001). In adults, intradermal tests revealed higher sensitivity, accompanied by larger area of skin reactions, regardless of type of venom. At concentrations lower than 0.1 µg/mL, 16% of wasp venom allergic children and 39% of bee venom allergic children had positive intradermal test results. The median tryptase level was significantly higher in adults than in children for the entire study group (P = .002), as well as in bee (P = .002) and wasp venom allergic groups (P = .049). CONCLUSION: The basic diagnostic profile in severe HVA reactors is age dependent. Lower skin test reactivity to culprit venom in children may have practical application in starting the intradermal test procedure with higher venom concentrations.

Antithrombin III as the Indicator of L-Asparaginase Activity in Children Treated for Acute Lymphoblastic Leukemia.

L-asparaginase (ASP) is widely used in the treatment of acute lymphoblastic leukemia (ALL) in children. Monitoring its activity is necessary because of the risk of drug inactivation as the result of an immune reaction. Besides allergic reactions, another frequent side effect of ASP treatment is coagulopathy, especially deficiency of antithrombin III (ATIII). The aim of this study was to analyze the relationship between ASP and ATIII activities and the possibility of ATIII activity use in an indirect ASP activity assessment. ASP and ATIII activity was measured in 76 children with ALL treated according to the ALL IC BFM 2002 protocol. A correlation between ASP and ATIII activities was found (R=-0.43, P=0.0001). ROC curve analysis revealed some utility regarding the determination of ATIII in identifying patients with low or undetectable ASP activity (area under the curve=0.87 [95% confidence interval, 0.77-0.96], P<0.0001 and 0.93 [95% confidence interval, 0.85-1.0], P<0.0001, respectively). Higher ATIII activity is associated with a higher probability of a decline in ASP activity. Examination of ATIII activity cannot replace a direct determination of ASP activity, but in the case of unavailability of the direct test, it can be a helpful surrogate parameter of drug activity.

[Hypoglycemia due to antibodies to exogenous insulin in a patient with type 2 diabetes ­ a case report of treatment with new generation oral hypoglycemic agents].

Hypoglycemic syndromes associated with immune reactions against insulin are rare phenomena described predominantly in Asians. Steroid therapy, immunosuppression or plasmapheresis is often required. Case report: A 73-year-old White woman with a 20-year history of type 2 diabetes was admitted to hospital due to recurrent incidents of hypoglycemia that started several months after insulin initiation (lispro 75/25) and increased in severity over the next 5 years. They were accompanied by postprandial hyperglycemia up to 25 mmol/l. The patient's glycated hemoglobin (HbA1c) was 70 mmol/ mol (8.6%). During hypoglycemic episodes recorded serum C-peptide was 0.57-0.73 nmol/l (1.7-2.2 ng/ml), while insulin concentration exceeded 7000 pmol/l (1000 mIU/l). Surreptitious insulin administration was ruled out as was, based on diagnostic imaging, the presence of an insulin secreting tumor. Anti-insulin antibody (AIA) level measured by 125I-insulin binding method was 92.5% (normal < 8.2%). Hypoglycemic episodes occurred for four days after discontinuation of insulin therapy and then resolved completely. Good glycemic control was maintained with metformin, acarbose and dapagliflozin. Three months later dapagliflozin was replaced with vildagliptine due to poor tolerance of a SGLT-2 inhibitor. Patient's HbA1c was 54 mmol/mol (7.1%), total fasting insulin level 2577 pmol/l and AIA binding 85.9%. Over the next year the patient has not experienced hypoglycemia and maintained good glycemic control, as HbA1c level was 53 mmol/l (7.0%) and AIA binding 39.5%. Conclusions: In this rare case of a patient with diabetes and hypoglycemic syndrome related to AIA, we achieved a rapid and stable remission of hypoglycemia without immunosuppression. Good glycemic control, despite 20-year history of diabetes was achieved with oral hypoglycemic agents.

Concentrations of adipokines in children before and after hematopoietic stem cell transplantation.

Adipokines have multiple effects, including regulation of glucose metabolism, cell proliferation, inflammation, and angiogenesis. The aim of the study was to determine plasma concentrations of adiponectin, apelin, leptin, and resistin as well as soluble leptin receptor in pediatric hematopoietic stem cell transplantation (HSCT). The expression of genes encoding the studied peptides was measured using microarray technique. Plasma concentrations of tested peptides were measured before and after oral glucose tolerance test in children treated with HSCT (n = 38) and in healthy controls (n = 26). The peptides were measured before HSCT (pre-HSCT group; n = 38) and after a median of 6 months after HSCT (post-HSCT group; n = 27 of 38 children treated with HSCT). In addition, measurements of fasting plasma glucose, insulin, lipids, and high-sensitivity C-reactive protein (hsCRP) were performed. In both HSCT groups, atherogenic lipid profile, low-grade systemic inflammation was observed. Leptin, adiponectin, and resistin also appear to be good markers of disease burden and low-grade systemic inflammation. Adipokines may be good markers of disease burden and may influence metabolic complications of HSCT. Future studies on larger groups of patients will explain if changes of the concentrations of leptin, adiponectin, and apelin observed in our study and confirmed by expression levels influence engraftment and reconstitution of cell lines.

[Fatty acid profile in phospholipids fraction with regard to blood pressure control in the kidney transplant patients].

Introduction: Hypertension is diagnosed in over 50% of renal transplant patients, and its presence is associated with significantly increased risk of cardiovascular complications. The pathogenesis of hypertension in this group of patients is complex. The use of immunosuppressive drugs is among the most important risk factors due to their hypertension-inducing properties. It is also suspected, that fatty acids may play an important role in the development of the disease; however, the mechanism of its formation has not yet been fully elucidated. Aim: Evaluation of fatty acid profile in phospholipids fraction in the blood serum of patients after renal transplantation, with respect to the blood pressure and immunosuppressive drug used. Materials and methods: The study included 65 kidney transplant patients treated with immunosuppressants: 24 patients received CsA (9 women, 15 men) and 41 were treated with Tac (15 women, 26 men). Blood pressure level was estimated by taking an average value of the last 3 measurements in an outpatient setting. Blood pressure exceeding 140/90 mmHg was considered as abnormally high. The number of antihypertensive drugs administered was based on the analysis of patients' records and direct interviews with patients. The concentration of each fatty acid was determined using gas chromatography. Statistical analysis was performed using Statistica 10. Results: There were no statistically significant differences between both: systolic blood pressure (136.16 vs. 133.31 mmHg, p=0.499) and diastolic blood pressure (80.62 vs 80.76; p=0.962) in patients treated with CsA compared to those taking Tac. Patients treated with CsA, received ­ on average ­ more antihypertensive drugs, compared to the group of patients taking Tac (2.64 vs 2.17), but this difference was also not statistically significant (p=0.174). The profile of fatty acids in renal transplant recipients treated with CsA was similar in both with normal blood pressure and with elevated blood pressure. In case of patients treated with Tac, significantly lower concentrations of C14 (p=0.015), C16:1 (p=0.039), C18:1 (p=0.043) and MUFA (monounsaturated fatty acids; p=0.049) were found in patients with higher values of blood pressure, compared to the ones whose blood pressure was within the norm. A significantly higher concentration of fatty acids: C14 (p=0.029), C16:1 (p=0.049) and C20:5 (p=0.029) was found in patients with normal blood pressure, treated with Tac as compared with treated with CsA. In contrast, regardless to the immunosuppressive drug used, no statistically significant differences were found between the patients' groups with elevated blood pressure values. Conclusion: The prevalence of hypertension in renal transplant recipients treated with CsA and Tac is similar. The fatty acid profile in phospholipids fraction depends on the blood pressure and the immunosuppressant used.

Enhanced liver fibrosis (ELF) test in obese children with ultrasound-proven liver steatosis.

OBJECTIVE: Non-alcoholic fatty liver disease (NAFLD) in obese children is a diagnostic challenge. The enhanced liver fibrosis test (ELF) based on the combination of serum concentration of hyaluronic acid (HA), aminoterminal propeptide of type III procollagen (PIIINP), tissue inhibitor of matrix metalloproteinase type 1 (TIMP-1) was developed as a noninvasive diagnostic tool for estimation of degree of liver fibrosis. The aim of our study was to investigate the performance of ELF test in obese children with ultrasound-proven steatosis in order assess the possibility of early detection of fibrotic changes in liver structure. MATERIAL AND METHODS: 58 obese (BMI >95th percentile) children, 27 male (mean age 13.9±2.65 years) and 31 female (mean age 13.82±2.64 years). Based on the liver ultrasound (US) examination results two groups of obese children were studied: group with steatosis (N=20, 8/12 M/F, mean age 14.2±1.90 years, BMI 32.9±5.60 kg/m2) and group with normal liver US (n=38, 19/19 M/F, mean age 13.7±2.94 years, BMI 30.4±4.67 kg/m2). Serum activity of aminotransferases (AST, ALT) and lactate dehydrogenase (LDH), and γ-glutamyl transpeptidase (GGT), and ELF test (HA, PIIINP, and TIMP-1 were analyzed. RESULTS: Children with liver steatosis presented with significantly higher AST (34.1 vs. 25.6 U/L), ALT (43.4 vs. 32 U/L), LDH (427.5 vs. 361.3 U/L), GGT (30.7 vs. 18.9 U/L). The ELF test value was also significantly higher in that group (8.98 vs. 8.49). Nevertheless no combination of measured parameters with ELF test value show better diagnostic value for differentiation between children with and without steaotosis. CONCLUSION: ELF test cannot be used for assessment of steatosis in obese children.

[Do we successfully treat anemia and calcium-phosphate disorders in children with chronic kidney disease at the beginning of the twenty-first century?]. / Czy na poczatku XXI wieku skutecznie leczymy niedokrwistosc i zaburzenia wapniowo-fosforanowe u dzieci z przewlekla choroba nerek?

UNLABELLED: In children with chronic kidney disease (CKD) anemia and calcium-phosphate disturbances are already present at early stages of the disease and require a comprehensive treatment. The aim of this study was to evaluate the efficacy of the treatment of biochemical disturbances, depending on the severity of CKD in children. MATERIAL AND METHODS: The study included 71 children (44 boys, 27 girls) with CKD stage 1-5. Mean age was 11 ± 5 years, mean height: 135.7 ± 28 cm and mean eGFR 32 ml/min/1.73 m2. The serum hemoglobin, urea, creatinine, cystatin C, calcium, phosphorus and parathyroid hormone (PTH) levels were measured. eGFR was calculated according to Schwartz and Filler formulas, employing creatinine and cystatin C as markers. Patients were divided into groups depending on the stage of CKD [group 1: CKD stage 1+2 (GFR > 60), group 2: CKD stage 3 (GFR = 30-59) Group 3: CKD stage 4 (GFR = 15-29 ml/min/1.73 m2), group 4 - dialyzed children]. RESULTS: The concentration of he- moglobin depending on the stage of CKD (group 1 vs. group 2 vs. group 3 vs group 4) was 12.95 vs. 12.68 vs. 12.47 vs. 11.3 g/dI, respectively. The concentration of total and ionized calcium was significantly lower in children on dialysis compared to patients treated conservatively. With the progression of CKD the concentration of phosphorus (1.39 vs. 1.4 vs. 1.49 vs. 1.82 mmolI) and PTH (21.7 vs 48.6 vs 99.9 vs. 219 pg/ml) significantly increased. Treatment with erythropoietin was used in 48% of children, calcium carbonate in 55% and alphacalcidol in 56% of patients. CONCLUSIONS: Despite the use of regular treatment, with the progression of CKD a progression of anemia, increased serum phosphate and parathyroid hormone and a decrease in calcium levels in studied children was observed. The severity of metabolic disorders in dialyzed children indicates the need for administration of new and more effective drugs, to prevent early enough complications of CKD in the form of mineral bone disease and cardiovascular complications.

[Blood volume for biochemistry determinations--laboratory needs and everyday practice]. / ObjetoSc krwi pobieranej do badan biochemicznych--potrzeby laboratorium a codzienna praktyka.

UNLABELLED: Blood loss due to diagnostic phlebotomy jest a very serious problem, especially for newborn, infants and critically ill patients on intensive care units. Although single blood loss can be easily tolerated in adults, in small babies and in patients who are frequently monitored based on laboratory tests iatrogenic anaemia can occur. AIM OF THE STUDY: To evaluate the blood volume drawn for routine biochemistry tests in relation to patient age and the number of parameters requested. MATERIAL AND METHODS: Blood volume drawn for routine biochemistry measurements from patients hospitalized in University Children's Hospital (N = 2980, children age from one day to 18 years) and in University Hospital (N = 859, adults, aged > 1.8 years) in Cracow has been analyzed. Blood volume was calculated based on regular tube diameter and blood heights in the tube. In case of microvettes the blood volume was 0.2 ml. Statistical analysis has been performed by using PRISM 5.0. The statistical significance was set at p < 0.05. RESULTS: The mean values of blood volume were 3.02 +/- 0.92 ml and 4.12 +/- 0.68 ml in children and adults, respectively. Analyzing blood volume drawn in children using both microvettes and regular tubes, significant correlation between blood volume and patient age (p < 0.001) as well the number of requested parameters (p < 0.001). The latest relationship was true only for up to five parameters. However, analyzing the blood volume drawn into only into regular tubes blood volume was not related to patients age and number of laboratory tests requested. The proportion of microvettes used for blood collection was highest for newborns and infants, and in all cases where only one to three laboratory tests were requested. CONCLUSIONS: 1. All educational programs for nurses and doctors should include the information about current laboratory automation and methods miniaturization; 2) The amount of blood volume needed by laboratory for the requested number of tests should always be taken into account when diagnostic phlebotomy is necessary.

24-hour urine metanephrine excretion in patients diagnosed with adrenal incidentaloma: impact of commonly used drugs on a clinical decision.

INTRODUCTION: Incidentaloma is an adrenal tumor detected during diagnostic imaging performed for extra­adrenal causes. Evaluation of metanephrine concentrations in a 24­hour urine collection can be a significant challenge in patients with multiple medications and comorbidities. OBJECTIVES: The aim of this study was to evaluate the effect of commonly used groups of drugs on metanephrine levels in the 24­hour urine collection. PATIENTS AND METHODS: A total of 1051 patients with adrenal mass below 10 Hounsfield units on unenhanced computed tomography were included in the study. Patients diagnosed with Cushing or Conn syndrome, adrenal carcinoma, pheochromocytoma, active extra­adrenal malignant neoplasms, and exacerbation of severe illnesses were excluded. Metanephrine, normetanephrine, and 3­methoxytyramine in the 24­hour urine collection were measured by high­performance liquid chromatography with electrochemical detection. Information on concomitant medication (ß­blockers, calcium channel blockers [CCBs], loop diuretics, thiazide diuretics, potassium­sparing diuretics, α­blockers, angiotensin­converting enzyme inhibitors / angiotensin II receptor blockers, metformin, nonmetformin antidiabetic drugs [NMADs], lipid­lowering drugs, proton pump inhibitors, levothyroxine, thyreostatics, antidepressants, neuroleptics, benzodiazepines, glucocorticosteroids, inhaled B­receptor agonists, and ipratropium) was collected from each patient. RESULTS: The urinary excretion of normetanephrine was significantly higher in the patients on ß­blockers, CCBs, loop diuretics, α­blockers, NMADs, and neuroleptics. α­Blockers increased urine metanephrine concentration, and NMADs, antidepressants, and glucocorticosteroids lowered it. There was no association between the analyzed drugs and urinary 3­methoxytyramine level. CONCLUSIONS: Many drug groups interfere with the measurement of urinary fractionated metanephrines. These interactions should be taken into account during interpretation of a hormonal evaluation, as they can be crucial for further management, especially for making a decision on surgical treatment.

Telocytes: new insight into the pathogenesis of gallstone disease.

The major mechanisms of gallstone formation include biliary cholesterol hypersecretion, supersaturation and crystallization, mucus hypersecretion, gel formation and bile stasis. Gallbladder hypomotility seems to be a key event that triggers the precipitation of cholesterol microcrystals from supersaturated lithogenic bile. Telocytes, a new type of interstitial cells, have been recently identified in many organs, including gallbladder. Considering telocyte functions, it is presumed that these cells might be involved in the signalling processes. The purpose of this study was to correlate the quantity of telocytes in the gallbladder with the lithogenicity of bile. Gallbladder specimens were collected from 24 patients who underwent elective laparoscopic cholecystectomy for symptomatic gallstone disease. The control group consisted of 25 consecutive patients who received elective treatment for pancreatic head tumours. Telocytes were visualized in paraffin sections of gallbladders with double immunofluorescence using primary antibodies against c-Kit (anti-CD117) and anti-mast cell tryptase. Cholesterol, phospholipid and bile acid levels were measured in gallbladder bile. The number of telocytes in the gallbladder wall was significantly lower in the study group than that in the control group (3.03 ± 1.43 versus 6.34 ± 1.66 cell/field of view in the muscularis propria, P < 0.001) and correlated with a significant increase in the cholesterol saturation index. The glycocholic and taurocholic acid levels were significantly elevated in the control subjects compared with the study group. The results suggest that bile composition may play an important role in the reduction in telocytes density in the gallbladder.

Blood loss from laboratory diagnostic tests in children.

BACKGROUND: Excessive diagnostic phlebotomy in children and critically ill patients is a frequent phenomenon in many hospitals. However, little attention is paid to a single blood volume taken routinely everyday from thousands of patients worldwide. The objective of the present study was to assess the volume of a single blood sample draw for laboratory testing in a pediatric population in relation to child age and weight, number of diagnostic tests requested by physicians, laboratory needs, and size of collection tube. METHODS: A single blood volume draw for diagnostic tests was measured in 3136 consecutive routine samples taken from children (from 1 day to 18 years old) and placed into a Microvette® or regular sampling tubes. The serum excess was calculated by taking into account the serum volume needed for the requested number of tests and the dead volume of analyzer. RESULTS: A huge variation in blood volume draws between individual patients, regardless of the number of tests requested, has been observed. The number of blood samples placed into the microvette decreased with patients' age, with 53.9% in children younger than 1 month old and 9.3% in children older than 12 years old. There was a clear-cut increase in the mean value of the blood volume draw with an increase in children's age. Only 2% of children up to 3 years old had a blood volume draw >1 mL/kg body weight. CONCLUSIONS: Each pediatric laboratory should have a clear-cut recommendation on the amount on blood volume necessary for the requested number of tests.

The accuracy of serum osmolarity calculation in small children.

Background: Serum osmolality can be measured (Omeas) or calculated (Ocal). Many formulas for Ocal have been already published, but data regarding the most accurate equation in small babies is not available. Thus, we aim to compare Omeas and Ocal obtained by different formulas in newborns and small children. Methods: The study included 280 serum samples taken from children, from the first day of life to 2 years (mean age 8.2 ± 7.6 months) treated in the University Children's Hospital in Krakow. The serum osmolality was measured by osmometer and calculated by 8 common formulas. Results: The mean value of Omeas (2 8 5 .8 ± 5 .1 mOsm/kgH2O) was significantly different as compared to the mean values of Ocal (p< 0.01) for all formulas, except Ocal obtained by the formula: 1.86*(N a + K) +1.15*Glu + Urea + 14. According to Bland-Altman analysis, this formula showed the best performance for estimating osmolality. In children under 3 months of life Passing-Bablok regression indicated both systematic and proportional error for results obtained by each formula compared to the measured values. Conclusions: To calculate osmolarity in children aged between 3 months and 2 years old the following equation: 1 .86*(N a + K) + 1.1 5*G lu+ U rea+ 14 might be used, whereas serum osmolality in children up to 3 month of life should be measured.

Biliary Amino Acids and Telocytes in Gallstone Disease.

The role of amino acids in cholesterol gallstone formation is not known. Therefore, the aim of the study was to determine the amino acid profile in the bile of patients with and without cholecystolithiasis in relation to bile lithogenicity and telocyte numbers within the gallbladder wall. The study included 23 patients with cholecystolithiasis and 12 gallstone-free controls. The levels of free amino acids in the bile were measured, and telocytes were identified and quantified in the gallbladder muscle wall. The mean values of valine, isoleucine, threonine, methionine, phenylalanine, tyrosine, glutamic acid, serine alanine, proline and cystine were significantly higher in the study group than in the controls (p from 0.0456 to 0.000005), and the mean value of cystine was significantly lower in patients with gallstone disease than in the controls (p = 0.0033). The relationship between some of the amino acids, namely alanine, glutamic acid, proline, cholesterol saturation index (CSI) and the number of telocytes was significant (r = 0.5374, p = 0.0051; r = 0.5519, p = 0.0036; and r = 0.5231, p = 0.0071, respectively). The present study indicates a potential relationship between the altered amino acid composition of bile and the reduced number of telocytes in the gallbladder muscle wall in cholelithiasis.

Amino acid profile in overweight and obese prepubertal children - can simple biochemical tests help in the early prevention of associated comorbidities?

Background: It is accepted that plasma branched-chain amino acids (BCAAs) and aromatic amino acids (AAAs) are closely related to metabolic risk. Arterial hypertension, metabolic syndrome, endothelial dysfunction, inflammation, and metabolic dysfunction-associated fatty liver disease (MAFLD) are frequently seen in obese patients. Many attempts have been made to find biochemical indicators for the early detection of metabolic complications in children. It is not known if different amino acid profiles and BCAA and AA concentrations in overweight and obese children correlate with chemerin, proinflammatory, and simple biochemical markers. Thus, the study aimed to find out the early markers of cardiovascular disease and MAFLD in overweight and obese children. Materials and methods: The study included 20 overweight and obese children (M/F 12/8; mean age 7.7 ± 2.3 years; BMI 26.8 ± 5.0 kg/m2) and 12 non-obese children (control group) (M/F 4/8; mean age 6.5 ± 2.2 years; BMI 14.8 ± 1.5 kg/m2). The following plasma amino acids were measured: aspartic acid, glutamic acid, serine, asparagine, glycine, glutamine, taurine, histidine, citrulline, threonine, alanine, arginine, proline, tyrosine, methionine, valine, isoleucine, leucine, phenylalanine, tryptophan, ornithine, and lysine. Chemerin, high-sensitivity C-reactive protein (hs-CRP), interleukin-6 (IL-6), and basic biochemistry parameters were measured. Results: The mean plasma levels of leucine, isoleucine, valine, phenylalanine, tyrosine, glutamic acid, and alanine were significantly higher in overweight and obese children than in the control group (p<0.03-p<0.0004). Conversely, the mean values of serine, asparagine, glutamine, and citrulline were significantly lower in overweight and obese children than in the control group (p<0.03-p<0.0007). Isoleucine, leucine, valine (BCAAs) tyrosine, and phenylalanine (AAAs) levels showed a positive correlation with uric acid, ALT, hs-CRP, and chemerin (r=0.80-0.36; p<0.05-p<0.00001), but not with IL-6. The mean values of glucose, IL-6, hs-CRP, chemerin, uric acid, and ALT were significantly higher in overweight and obese children than in the control group (p<0.03-p<0.00002). In contrast, the lipid profile did not differ between groups. Conclusion: An abnormal amino acid profile in overweight and obese pre-pubertal children, accompanied by elevated ALT and UA observed in the studied cohort, may suggest early metabolic disturbances that can potentially lead to metabolic syndrome, or MAFLD, and increased cardiovascular risk.