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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.

Background: Monitoring of bilirubin is essential during early neonatal life. Bilirubin in high concentration is toxic to the brain and might cause irreversible neurological damage. Several different methods for bilirubin determination are available nowadays, but inconsistent results may be obtained. The study aimed to compare dry chemistry methods with vanadate oxidation method for bilirubin determination in relation to hematocrit and albumin level in neonates and infants. Methods: The study included 98 consecutive serum samples from newborns and infants (47 boys and 51 girls, mean age 19 ± 15 days) treated in the University Children's Hospital in Krakow. Total bilirubin (TBil) and neonatal bilirubin (NBil) concentration were measured by dry chemistry analyser (Vitros 4600, Ortho Clinical Diagnostics Inc.). Total bilirubin (TBilV) was also measured using vanadate oxidation method (Cormay, Poland). Albumin concentration and blood morphology have been routinely determined in all children. Results: No significant differences between the mean value of NBil (69.00 ± 67.76 mmol/L), TBil (81.26 ± 70.13 mmol/L) and TBilV (75.90 ± 60.62 mmol/L) were noticed. High coefficient correlation between NBil and TBil as well as between NBil and TBilV were noticed (Pearson's analysis, r = 0.99, r = 0.97, respectively; p < 0.0001 in both cases). There was a positive correlation between the difference (TBilV - NBil) and hematocrit (p < 0.009, r = 0.2664). Conclusions: In newborns and infants the same method for bilirubin determination should be used when the concentration of bilirubin is monitored. When using vanadate oxidation method for bilirubin determination, hematocrit value should be taken into account when results are interpreted.

Background: Polyunsaturated fatty acids (PUFAs) play a role in the development/progression of colon cancer. The aim of the study was to assess the relation between serum phospholipids PUFAs, colorectal tumour localization and disease progression. Methods: A total of 67 patients (18 with proximal colon, 17 with distal colon and 32 with rectal tumour localization) as well as 16 controls were studied. One year after surgery, 33 patients had disease progression. Serum levels of C16:1(n-7), C18:1(n-9), C18:3(n-3), C20:5(n-3), C22:6(n3), C18:2(n-6), C20:2(n-6), C20:4(n-6) fatty acids of serum phospholipids were quantitatively measured before surgery by gas-chromatography. Results: Significantly higher mean value of C 18:2, as compared to control, has been noted only for patients with proximal (p< 0.05) and distal tumour (p< 0.03) localization. The lower mean level of C 20:5 and unsaturation index (UI) were observed in colorectal cancer patients regardless the tumour localization, but the statistical difference was noted only for patients with proximal tumours (p< 0.05, p

Introduction: Incidentaloma is an adrenal tumor detected during diagnostic imaging performed for extra‑adrenal causes. Evaluation of metanephrine concentrations in a 24‑hour 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 24‑hour 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 extra‑adrenal malignant neoplasms, and exacerbation of severe illnesses were excluded. Metanephrine, normetanephrine, and 3‑methoxytyramine in the 24‑hour urine collection were measured by high‑performance liquid chromatography with electrochemical detection. Information on concomitant medication (β‑blockers, calcium channel blockers [CCBs], loop diuretics, thiazide diuretics, potassium‑sparing diuretics, α‑blockers, angiotensin‑converting enzyme inhibitors / angiotensin II receptor blockers, metformin, nonmetformin antidiabetic drugs [NMADs], lipid‑lowering drugs, proton pump inhibitors, levothyroxine, thyreostatics, antidepressants, neuroleptics, benzodiazepines, glucocorticosteroids, inhaled B‑receptor agonists, and ipratropium) was collected from each patient., Results: The urinary excretion of normetanephrine was significantly higher in the patients on β‑blockers, CCBs, loop diuretics, α‑blockers, NMADs, and neuroleptics. α‑Blockers increased urine metanephrine concentration, and NMADs, antidepressants, and glucocorticosteroids lowered it. There was no association between the analyzed drugs and urinary 3‑methoxytyramine 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.

Antibiotic therapy requires appropriate dosage of drugs for effective treatment. Too low antibiotic concentrations may lead to treatment failure and the development of resistant pathogens, whereas overdosing may cause neurological side effects or hemolytic diseases. Meropenem and linezolid are used only in the treatment of serious infections or when other antibiotics are no longer effective as well as for treating central nervous system infections. It is difficult or sometimes even impossible to predict the relation between dosing of antibiotics and its cerebrospinal fluid (CSF) concentration; thus, a method of determining antibiotics not only in the blood but also in the CSF is needed. Analytical method validation is an integral part of good laboratory practice and ensures high accuracy of the results. We performed complete validation process according to the Food and Drug Administration and European Medicine Agency, covering the aspects precision, specificity, accuracy, recovery, limit of detection, limit of quantification, stability, carry-over, and matrix effects. Our liquid chromatography-tandem mass spectrometry method for the simultaneous measurement of meropenem and linezolid in different matrix meets all the acceptance criteria. The method was successfully applied to determine meropenem and linezolid concentrations in serum and CSF samples obtained from children treated with these antibiotics.

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/m 2 ) and 12 non-obese children (control group) (M/F 4/8; mean age 6.5 ± 2.2 years; BMI 14.8 ± 1.5 kg/m 2 ). 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., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Bugajska, Berska, Wójcik and Sztefko.)

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.

Introduction: The aim of the study was to address the fatty acid (FA) status and its relationship with disease activity in patients with inflammatory bowel disease (IBD)., Methods: FA levels of the phospholipid fraction in serum and a colon biopsy specimen were measured in 17 patients with IBD., Results: A negative correlation between the histological activity of inflammation of the disease and the ratio of polyunsaturated FAs/no polyunsaturated FAs was observed. Moreover, the level of that ratio was lower in patients with IBDs as compared to controls., Conclusions: The FA profile in serum and in a colon biopsy specimen in patients with IBD is characteristic for essential fatty acid insufficiency., Competing Interests: The authors declare no conflict of interest., (Copyright: © 2022 Termedia & Banach.)

Introduction: The influence of growth hormone (GH) treatment on amino acids (AAs) profile in patients with Turner syndrome (TS) was investigated., Material and Methods: The study group included girls with TS: treated with GH (GH+) and girls with no GH treatment (GH-). The control group consisted of healthy girls. Free plasma AAs were measured by the LC/MS/MS., Results: The plasma concentrations of glutamine, threonine were significantly higher in group GH+ than in group GH- (p < 0.05). In group GH- the values of glutamine, alanine, isoleucine, glutamic acid were significantly different than in the control (p < 0.05-p < 0.008)., Conclusion: AAs profile in girls with TS might be characteristic for the disease but also depends on GH treatment.

Girls with Turner syndrome (TS) are at increased risk of developing insulin resistance and coronary artery disease as a result of hypertension and obesity frequently seen in these patients. On the other hand, it is known that obesity is associated with increased serum levels of branched-chain amino acids (BCAAs: valine; leucine and isoleucine) and aromatic amino acids. The aim of the study is to compare the metabolic fingerprint of girls with TS to the metabolic fingerprint of girls with obesity. Metabolic fingerprinting using an untargeted metabolomic approach was examined in plasma from 46 girls with TS (study group) and 22 age-matched girls with obesity (control group). The mean values of BCAAs, methionine, phenylalanine, lysine, tryptophan, histidine, tyrosine, alanine and ornithine were significantly lower in the study group than in the control ( p from 0.0025 to <0.000001). Strong significant correlation between BCAAs, phenylalanine, arginine, tyrosine, glutamic acid, citrulline and alanine, and body mass index expressed as standard deviation score BMI-SDS in the patients with obesity ( p from 0.049 to 0.0005) was found. In contrast; there was no correlation between these amino acids and BMI-SDS in the girls with TS. It is suggested that obesity in patients with TS is not associated with altered amino acids metabolism.

Introduction: Gallstone disease is associated with insulin resistance, type 2 diabetes mellitus, and increased risk of incident ischemic heart disease. It is known that the profile of branched‑chain amino acids (BCAAs) is altered in cardiac diseases as well as metabolic diseases, such as diabetes and obesity. The role of BCAAs in gallstone disease is still not known., Objectives: The aim of this study was to evaluate the concentration of essential amino acids and incretin hormones in patients with cholecystolithiasis., Patients and Methods: The study included 31 patients with cholecystolithiasis and 25 gallstone‑free controls. The levels of free exogenous and endogenous amino acids, bile acids, glucagon‑like peptide 1, glucose‑dependent insulinotropic polypeptide, ghrelin, C-peptide, and insulin were measured in the fasting state and 1 hour after consumption of a 300‑kcal mixed meal., Results: The mean fasting and postprandial levels of valine, isoleucine, leucine, and lysine were higher in the study group than in controls (all P.

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.

Objectives: It has been suggested that some amino acids are involved in the pathogenesis of autistic disorders. The aim of the study was to evaluate the plasma amino acids profile in young males with autism., Methods: Total of 27 autistic boys (aged 2-10 years, the study group) without any metabolic disorders and 13 healthy boys (aged 2-9 years, control group) were included in the study. In all subjects fasting blood plasma free amino acids (both exogenous and endogenous) were quantitatively measured by high performance liquid chromatography with UV-VIS detection., Results: The mean plasma concentration values of citrulline, .-aminobutyric acid, isoleucine, leucine, phenylalanine, tryptophan and ornithine were significantly lower in boys with autism as compared to the control group (p < 0.03, p < 0.04, p < 0.02, p < 0.02, p < 0.05, p < 0.02, p < 0.05, respectively). The areas under the Receiver Operating Characteristic curves for these amino acids ranged from 0.637 to 0.726. None of the amino acids measured differentiate autistic children from healthy children. The sum of exogenous amino acids was lower in the study group than in the control group but this difference was not statistically significant., Conclusions: Lower levels of exogenous amino acids confirm the possible role of these amino acids in autism. Determination of exogenous amino acids in plasma, however, cannot be used as a diagnostic test but it can still support autistic patients care.

Background: One of the strongest risk factors of cardiovascular disease is a high concentration of low-density lipoprotein cholesterol (LDL-C); thus, the accurate measurement of LDL-C concentration in children is important. The aim of this study was to compare the concentration of LDL-C measured by direct method and the LDL-C concentration estimated by different formulas with the modified β-quantification method in children with type 1 diabetes., Methods: Thirty-one serum samples received from diabetic children (15/18; M/F) and 26 serum samples taken from nondiabetic children (18/14; M/F) were used in the study. LDL-C concentrations were determined by direct enzymatic method (LDL-CD) and the modified β-quantification method (LDL-CmBQ). The concentration of LDL-C was also calculated by different formulas., Results: The results of LDL-CmBQ and LDL-CD correlated. However, the correlation coefficient obtain in the nondiabetic samples was much higher (r=0.927; p<0.001) than the correlation coefficient for LDL-CmBQ and LDL-CD concentration obtained in the diabetic samples (r=0.691; p<0.001). Additionally, the coefficients of correlation between the LDL-CmBQ concentration and the LDL-C concentrations calculated by different formulas were lower in diabetic (range: 0.514-0.693) than in nondiabetic samples (range: 0.834-0.937). Bland-Altman plots showed much higher confidence intervals for 95% limits of agreement for the differences between LDL-CmBQ and LDL-CD as well as for the differences between LDL-CmBQ and LDL-C estimated by different formulas for samples from the diabetic samples compared with nondiabetic samples., Conclusions: The limitations of methods measurement and of each of the formulas should be taken into account when a medical decision to lower LDL-C is made in the treatment of diabetic children.

Aim: Estimation of eGFR in children with normal kidney function using the Schwartz equations results in underestimating real GFR., Materials and Methods: We propose modification of three Schwartz equations - two based on creatinine concentration (eGFRScrBS bedside) and (eGFRScr) and one 3-marker based on creatinine, urea and cystatin C concentrations (eGFRS3M). The iohexol test (reference method) was performed 417 times in 353 children >2 years with mean GFR: 98 ± 31.6 ml/min/1.73m(2). The assessment included also the Filler and Zappitelli equations. The modification was performed using methods: (1) based on equation, eGFRcor = a [eGFR - T] + T, where T = 50, if eGFR > T, and a equals for: eGFRScrBS 1.4043, for eGFRScr 2.0048, for eGFRS3M 1.2951, and (2) based on correction of all coefficients of the original equation., Results: For comparison of all the results and for children with GFR< 60, 60-90, 90-135 and > 135 ml/min/1.73m(2) the correlation coefficient, relative error (RE) and root mean square relative error (RMSRE) was employed and revealed improvement of RE from 25.9 to 6.8 and 3.9% (depending on the correction method) for eGFRScr; from 19 to 8.1 and 3.9% for eGFRScrBS and: from 11.6% to 2.0 and 2.3% for eGFRS3M (respectively). The RMSRE values changed from 30 to 21.3 and 19.8% for eGFRScr, from 25.1 to 21.6 and 19.8% for eGFRScrBS and from 19.1 to 15.8 and 15.3 % for eGFRS3M., Conclusions: Modifications of Schwartz equations at GFR > 60 ml/min/1.73m(2) significantly improves the accuracy of calculating eGFR. The 3-markers equation is more accurate and should be employed frequently.

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.

Purpose: We assessed the reliability of calculating eGFR in children as compared to the iohexol disappearance test (GFR-I), which was performed 417 times in 353 children aged 2 and more., Material/methods: eGFR was estimated with equations based on serum creatinine: Schwartz (1: eGFR-Scr), Cockroft-Gault (2: eGFR-CG) and MDRD (3: eGFR-MDRD), and on creatinine clearance (4: eGFR-U), or relying on serum cystatin C: Hoeck (5: eGFR-H), Bokenkamp (6: eGFR-B) and Filler (7: eGFR-F), and on the three Schwartz markers (8: eGFR-S3M). Mean relative error (RE), correlation (R), Bland-Altman analysis and accuracy of GFR-I were studied in all patients and in subgroups: at GFR<60ml/min/1.73m(2); in children aged ≤12 and >12., Results: The results by eGFR-Scr, eGFR-S3M demonstrated no statistical difference to GFR-I at GFR<60ml/min/1.73m(2), but underestimated eGFR at higher filtration values by 11.6±15.1% and 19.1±16.4, respectively (p<0.0000). The eGFR-B, eGFR-F and eGFR-MDRD equations illustrated important overestimation of reference GFR results (RE: 84±44.2%; 29.5±27.9%, 35.6±62%; p<0.0000 for all). The MDRD and C-G formulas showed statistically better consistency in children aged >12. A good agreement was achieved by the eGFR-H equation (5.1±21.9%; p<0.0000; R=0.78)., Conclusions: (1) Schwartz equations show a good conformity at GFR<60ml/min/1.73m(2), but underestimate the results at higher GFR values. (2) The Bokenkamp equation with original coefficient should not be employed in children. (3) The use of the Hoeck formula in all children and C-G and MDRD formula in children aged >12 is possible. (4) The error of eGFR calculations increases at higher GFR values., (Copyright © 2014 Medical University of Bialystok. Published by Elsevier Urban & Partner Sp. z o.o. All rights reserved.)