Serum carbohydrate sulfotransferase 7 in lung cancer and non-malignant pulmonary inflammations.

BACKGROUND: Carbohydrate sulfotransferases (CHST) were shown to be involved in carcinogenesis. The aim of the study was to assess the diagnostic value of serum CHST7 concentration in differentiation between lung cancer and non-malignant pulmonary inflammations. METHODS: Clinical case-control study involving 125 participants was conducted: the control group containing cases of pneumonia and chronic obstructive pulmonary disease was compared to the lung cancer group composed of primary and metastatic cancers. Serum concentrations of CHST7 and routinely used markers including carcinoembryonic antigen (CEA), cytokeratin fragment 21-1 (CYFRA 21-1) and neuron-specific enolase (NSE) were determined for each participant using immunochemical methods. Statistical association, receiver operating characteristic (ROC) analysis and cross-validation were used for the evaluation of CHST7 either as a standalone biomarker or as a part of a biomarker panel. RESULTS: In comparison to the control group, serum CHST7 was elevated in lung cancer (p<0.001), but no differences between the overall stages of primary cancers were detected (p=0.828). The differentiation performance in terms of ROC area under curve (AUC) was 0.848 making CHST7 superior biomarker to the NSE (p=0.031). In comparison to CEA and CYFRA 21-1, the performance differences were not detected. CHST7 was not correlated to other biomarkers, and its addition to the routine biomarker panel significantly improved the cross-validated accuracy (85.6% vs. 75.2%) and ROC AUC (p=0.004) of the differentiation using a machine learning approach. CONCLUSIONS: Serum CHST7 is a promising biomarker for the differentiation between lung cancer and non-malignant pulmonary inflammations.

Differentiation of acute pyelonephritis from other febrile states in children using urinary neutrophil gelatinase-associated lipocalin (uNGAL).

BACKGROUND: Acute pyelonephritis is a severe disease which is sometimes difficult to recognize based on clinical symptoms and routinely available diagnostic tests, especially in young children. The aim of this study was to assess the diagnostic value of urinary neutrophil gelatinase-associated lipocalin (uNGAL) as a biomarker of acute pyelonephritis. METHODS: In this case-control study we analyzed 134 children (median age 2.5 years) who were admitted to the Pediatric Clinic of University Hospital Centre Osijek, Croatia. Eighty of them had acute pyelonephritis, while 54 children had febrile state of different etiology including cystitis and they represented the control group. uNGAL, white blood cells, C-reactive protein, urinanalysis, urine culture, kidney ultrasound and a dimercaptosuccinic acid scintigraphic scan were done for each child. uNGAL was measured using chemiluminiscent microparticle immunoassay on ARHITECT i1000SR (Abbott Diagnostics, IL, USA). RESULTS: uNGAL values were significantly higher in children with acute pyelonephritis compared to the control groups (113.6 ng/mL vs. 10.2 ng/mL, p<0.001). A receiver operating characteristic curve comparison was done for tested parameters and encouraging results were obtained for uNGAL (AUC=0.952). A cut-off value of 29.4 ng/mL had 92.5% sensitivity and 90.7% specificity. We showed that uNGAL can also serve in differentiating acute pyelonephritis from cystitis (cut-off 38.5 ng/mL), and for differentiation of cystitis from febrile states with etiology other than urinary tract infection (UTI) (cut-off 20.4 ng/mL). CONCLUSIONS: uNGAL can be a useful diagnostic biomarker in acute pyelonephritis in children, but also in differentiating cystitis from febrile states other than UTI.

VKORC1 gene polymorphisms and adverse events in Croatian patients on warfarin therapy.

OBJECTIVE: The objective was to determine the impact of VKORC1 polymorphisms on warfarin anticoagulant therapy (stable warfarin maintenance dose, time required to reach therapeutic dose and time spent in therapeutic range) and its adverse events (overanticoagulation and bleeding events, time to first overanticoagulation or bleeding event, and therapy for bleeding events) in Croatian patients. MATERIALS: Blood samples were collected from 186 patients on stable warfarin therapy. METHODS: VKORC1 1173C>T and VKORC1 –1639G>A gene polymorphisms were analyzed using real-time PCR. Prothrombin time international normalized ratio (INR) values were determined and overanticoagulation as well as bleeding events were recorded. RESULTS: Both tested VKORC1 gene polymorphisms (VKORC1 1173C>T and VKORC1 -1639G>A) were in perfect linkage disequilibrium. Genotype analysis showed that 33.9% of patients were homozygous for wild-type, 46.8% were heterozygous and 19.4% were homozygous for the variant allele. We have found a statistically significant difference between variantallele carriers and wild-type patients in stable warfarin maintenance dose (p<0.001) and incidence of bleeding events (p=0.040). Patients homozygous for variant-allele were more likely to experience an overanticoagulation event in the first 30 days of therapy (p=0.040). CONCLUSIONS: Our study showed that VKORC1 1173C>T and VKORC1 -1639G>A gene polymorphisms are associated with stable warfarin maintenance dose and adverse events of warfarin therapy.

Association of IL-1ß and IL-10 Polymorphisms with Prostate Cancer Risk and Grade of Disease in Eastern Croatian Population.

Single nucleotide polymorphisms (SNPs) in the promotor regions of cytokine genes included in angiogenesis may influence prostate cancer (PCa) development via regulation of the pathways of tumor angiogenesis. The aim of the present study was to investigate the association of IL-1 female +3954 (rs1143634) and IL-10-1082 (rs1800896) polymorphisms with PCa risk and aggressiveness in eastern Croatian patients. One hundred twenty PCa patients and 120 benign prostatic hyperplasia (BPH) controls were genotyped using real-time PCR (LightCycler Instrument, Roche Diagnostics) and the melting curve analysis method. There was no significant difference in the frequency of genotypes for the two polymorphisms between PCa patients and controls (Χ2 = 0.857, p = 0.355 for IL-female 1; Χ2 = 0.026, p = 0.872 for IL-10). Carriers of the IL-10-1082A>G variant were found to be associated with the Gleason score (GS) > 7 (AA versus GA+GG, OR = 3.47, 95% CI 1.11-10.88, p = 0.033). There was no significant difference in the frequency of genotypes for the two polymorphisms and the presence of metastatic disease in PCa patients. These results suggest that tested SNPs associated with differential production of IL-1 female and IL-10 are not risk factors for PCa and do not correlate with the presence of distant metastasis in eastern Croatians. We found that IL-10-1082 GA+/or GG carriers have a higher risk of developing PCa with GS > 7 in eastern Croatians.

Association study of cytochrome P450 1A1*2A polymorphism with prostate cancer risk and aggressiveness in Croatians.

Cytochrome P450 1A1 (CYP1A1) is an enzyme participating in the bioactivation of various endogenous and environmental reactive compounds that can bind to DNA and thus induce cancerogenesis. Gene encoding the enzyme is expressed in the prostate tissue and is polymorphic. CYP1A1*2A gene polymorphism is associated with elevated enzyme activity and/or inducibility which can lead to accumulation of genotoxic compounds and consequently to cancerogenesis. We examined the association of this polymorphism with prostate cancer (PCa) risk and aggressiveness. The case-control study consisted of 120 PCa patients and 120 benign prostatic hyperplasia (BPH) controls, in Croatian population. Regarding aggressiveness, PCa patients were grouped according to the Gleason score (GS), tumor stage (T) and existence of distant metastasis (M). The polymorphism was analyzed using real-time polymerase chain reaction (PCR). We did not observe association of mutated allele with PCa risk, neither with PCa aggressiveness. Furthermore, frequency of polymorphic genotype was slightly higher in BPH group (16.6% vs. 14.2%, respectively) and also in less aggressive form of PCa (20.4% vs. 9.6% for GS < 7; 15.6% vs. 9.1% for T < 3; 16.7% vs. 10.0% for no distant metastasis). Comparing our findings with other published results, we can assume that the ethnicity influence the genotype distribution and thus may affect the etiology of PCa, even possibly in the way to cause an opposite effect among different ethnic groups. Given the small number of participants, results should be validated on the larger sample size.

Comparison between clinical significance of serum proinflammatory protein interleukin-6 and classic tumor markers total PSA, free PSA and free/total PSA prior to prostate biopsy.

The aim of the study was to clarify whether serum levels of proinflammatory cytokine interleukin-6 (IL-6) could be a useful marker in prostate diseases. Serum IL-6 was determined prior to prostate biopsy procedure in 82 patients with prostate adenocarcinoma (PCa), 25 patients with benign prostatic hyperplasia (BPH), 24 patients with high-grade prostatic intraepithelial neoplasia (PIN) and 17 patients with chronic prostatitis. Serum IL-6 levels were compared with total PSA (tPSA), free PSA (fPSA) and the free/total ratio (f/tPSA) serum levels. Statistically significant difference was not found in serum IL-6 levels among the four groups (p = 0.088). However, the patients with poorly differentiated PCa with Gleason score (GS) 4 + 3 = 7 and > 7 had significantly higher serum IL-6 levels than the patients with moderately differentiated PCa with GS 3 + 4 = 7 and < 7 (p = 0.007). The findings suggest that serum IL-6 level might be a potentially useful marker for poorly differentiated PCa.

Iron Metabolism, Calcium, Magnesium and Trace Elements: A Review.

Iron (Fe) is fundamental to life on earth. In the human body, it is both essential and harmful if above threshold. A similar balance applies to other elements: calcium (Ca), magnesium (Mg), and trace elements including copper (Cu), zinc (Zn), lead (Pb), cadmium (Cd), mercury (Hg), and nickel (Ni). These elements share some proteins involved in the absorption and transport of Fe. Cu and Cd can inhibit Fe absorption, while excess of Fe may antagonize Cu metabolism and reduce ceruloplasmin (Cp). Excessive Fe can hinder Zn absorption and transferrin (Trf) can bind to both Zn and Ni. Ca is able to inhibit the divalent metal transporter 1 (DMT1) in a dose-dependent manner to reduce Fe absorption and low Mg concentrations can exacerbate Fe deficiency. Pb competitively inhibits Fe distribution and elevated Cd absorption reduces Fe uptake. Exposure to Hg is associated with higher ferritin concentrations and Ni alters intracellular Fe metabolism. Fe removal by phlebotomy in hemochromatosis patients has shown to increase the levels of Cd and Pb and alter the concentrations of trace elements in some types of anemia. Yet, the effects of chronic exposure of most trace elements remain poorly understood.

Cognitive phenomena measurement with time window-based multispectral brain mapping.

BACKGROUND: Cognitive neuroscience experiments require accurate and traceable methods of measuring cognitive phenomena, analyzing and processing data, and validating results, including measurement of impact of such phenomena on brain activity and consciousness. EEG measurement is the most widely used tool for evaluation of the experiment's progress. To extract more information from the EEG signal, continuous innovation is necessary to provide a broader range of information. OBJECTIVE: This paper presents a new tool for measuring and mapping cognitive phenomena using time window-based multispectral brain mapping of electroencephalography (EEG) signals. METHODS: The tool was developed using Python programming language and enables users to create brain maps images for six spectra (Delta, Theta, Alpha, Beta, Gamma, and Mu) of EEG signal. The system can accept an arbitrary number of EEG channels with standardized labels based on the 10-20 system, and users can select the channels, frequency bandwidth, type of signal processing, and time window length to perform the mapping. RESULTS: The key advantage of this tool is its ability to perform short-time brain mapping, which allows for the exploration and measurement of cognitive phenomena. The tool's performance was evaluated through testing on real EEG signals, and results demonstrated its effectiveness in accurately mapping cognitive phenomena. CONCLUSION: The developed tool can be used in various applications, including cognitive neuroscience research and clinical studies. Future work involves optimizing the tool's performance and expanding its capabilities.

Syndrome of inappropriate antidiuretic hormone secretion as an adverse reaction of ciprofloxacin: a case report and literature review.

Antidiuretic hormone (ADH) is secreted by the posterior pituitary gland. Unsuppressed release of ADH leads to hyponatremia. This condition is referred to as syndrome of inappropriate antidiuretic hormone secretion (SIADH). Hereby, a case report is presented on ciprofloxacin-induced SIADH. A 67-year-old male patient was examined in the emergency room with symptoms of lethargy, headache, lack of attention, and a generally depressed mood lasting for three days. One week prior, empirical antimicrobial therapy involving ciprofloxacin for prostatitis was initiated. Laboratory analysis showed no relevant abnormalities except for hyponatremia (Na = 129 mmol/L). Chronic hyponatremia, thyroid dysfunction, and adrenal dysfunction were ruled out. Serum osmolality was 263 mOsmol/kg, urine osmolality was 206 mOsmol/kg, and urine sodium was 39 mmol/L. Given that all criteria for SIADH were met, ciprofloxacin was discontinued, and fluid restriction was advised. Four days later, the patient's serum sodium concentrations nearly normalized (Na = 135 mmol/L), and all symptoms resolved. The Naranjo Scale yielded a score of 8, supporting the likelihood of a probable adverse reaction to ciprofloxacin. This case is presented to raise awareness among clinicians about the potential of ciprofloxacin to cause even mild hyponatremia.

GDF-15 Levels and Other Laboratory Findings as Predictors of COVID-19 Severity and Mortality: A Pilot Study.

Growth differentiation factor 15 (GDF-15) is a stress-induced cytokine associated with acute and chronic inflammatory states. This prospective observational study aimed to investigate the prognostic roles of GDF-15 and routine clinical laboratory parameters in COVID-19 patients. Upon the admission of 95 adult hospitalized COVID-19 patients in Croatia, blood analysis was performed, and medical data were collected. The patients were categorized based on survival, ICU admission, and hospitalization duration. Logistic regression and ROC curve methods were employed for the statistical analysis. Logistic regression revealed two independent predictors of negative outcomes: CURB-65 score (OR = 2.55) and LDH (OR = 1.005); one predictor of ICU admission: LDH (OR = 1.004); and one predictor of prolonged hospitalization: the need for a high-flow nasal cannula (HFNC) upon admission (OR = 4.75). The ROC curve showed diagnostic indicators of negative outcomes: age, CURB-65 score, LDH, and GDF-15. The largest area under the curve (AUC = 0.767, specificity = 65.6, sensitivity = 83.9) was represented by GDF-15, with a cutoff value of 3528 pg/mL. For ICU admission, significant diagnostic indicators were LDH, CRP, and IL-6. Significant diagnostic indicators of prolonged hospitalization were CK, GGT, and oxygenation with an HFNC upon admission. This study reaffirms the significance of the commonly used laboratory parameters and clinical scores in evaluating COVID-19. Additionally, it introduces the potential for a new diagnostic approach and research concerning GDF-15 levels in this widespread disease.

Vitamin K epoxide reductase complex 1 (VKORC1) gene polymorphisms in population of Eastern Croatia.

Vitamin K epoxide reductase (VKOR) is a key enzyme in the y-carboxylation of proteins associated with important bodily functions (coagulation, bone metabolism, etc.). This feature is particularly used in warfarin therapy, which blocks the VKOR enzyme and leads to production of dysfunctional coagulation proteins. Genetic factors, particularly vitamin K epoxide reductase complex 1 (VKORC1) gene, have greatest influence on warfarin therapy. The aim of this study was to determine the distribution of VKORC1 1173C > T and VKORC1 -1639G > A polymorphisms, which are most important for warfarin therapy. We investigated 420 unrelated healthy individuals, mostly blood donors, from region of the Eastern Croatia. Both investigated polymorphisms were in perfect linkage disequilibrium (LD) and showed identical results. 151 patients (36%) were homozygous for the wild-type (C/C and G/G), 196 (47%) were heterozygous (C/T and G/A) and 73 (17%) were homozygous for the variant allele (T/T and A/A). Number of normal alleles among individuals was 498 (59.3%), and number of variant alleles was 342 (40.7%). The data obtained are in good agreement with the results of studies in other European populations.

Lack of association between TNF-alpha promoter polymorphism and prostate carcinoma susceptibility in eastern Croatian population.

A single nuclear polymorphisms (SNPs) in the promoter region of the tumor necrosis factor-alpha (TNF-alpha) gene are involved in regulation of expression levels of TNF-alpha and therefore are associated with oncogenesis of several cancers. Aim of our study was to investigate the effect of G--->A polymorphism at -308 position in the promoter region of the TNF-alpha gene on prostate cancer (CalphaP) susceptibility in a subset of patients from Eastern Croatia. Study population consisted of 240 patients (120 with CalphaP, 120 controls). They were genotyped for TNF-alpha G-308A polymorphism using real-time PCR (LightCycler Instrument, Roche Diagnostics) and melting curve analysis method. X(2) test was used to compare distribution of TNF-alpha polymorphism genotypes between patients and control group. Relative risk was estimated by the odds ratio (OR). There was no significant statistical difference (X(2)=0.000, DF=1, p=1, OR=1, 95%CI=0.5537-1.8059) between patients and control group. Besides, data of CalphaP patients were stratified according to pathohistological diagnosis (PHD) by Gleason score and groups were compared according to TNF-alpha genotypes. Also, all patients and CalphaP patients were grouped according to prostate volume (V) into three groups: V<50 mL, V50-100 mL, V>100 mL. These groups were also compared according to TNF-alpha genotypes. There were no significant statistical differences between any of groups. Our findings suggest that TNF-alpha -308 SNP is not associated with CalphaP in Eastern Croatia population.

Interleukin-6 polymorphism and prostate cancer risk in population of Eastern Croatia.

Recent studies suggest that chronic inflammation is crucial in the development and progression of prostate cancer (CaP). Interleukin-6 (IL-6) is a proinflammatory cytokine that plays an important role in intraprostatic inflammation and thus carcinogenesis. The -174G > C polymorphism of IL-6 gene has been associated with high IL-6 producer phenotype and an increased risk for CaP. The aim of this study was to evaluate the association between the mentioned IL-6 polymorphism and CaP risk, as well as to compare the genotype frequency between the different tumour grades of CaP, in population of Eastern Croatia. We analyzed the IL-6 polymorphism in 120 CaP patients and 120 controls with benign prostatic hyperplasia (BPH). CaP patients and BPH controls did not statistically differ in studied IL-6 polymorphism. Furthermore, high IL-6 producer genotypes (GG or GC) were more frequent in controls than in CaP group (86.7% vs 80.8%, respectively, p = 0.147). Also, no statistically significant difference in IL-6 high and low producer genotype frequency was noticed between well, moderately and poorly differentiated tumours. Our results, taken together with other studies on the subject, suggest that IL-6 - 174 single nucleotide polymorphism (SNP) distribution may differ between various ethnic groups and that a single cytokine gene polymorphism has probably just a minor effect on CaP susceptibility. Further studies should be performed to clarify the link between SNPs of different cytokines and the risk for CaP.

SARS-CoV-2 and Plasmodium falciparum coinfection: a case report.

BACKGROUND: A rare case of coinfection of Plasmodium falciparum and SARS-CoV-2 disease in Croatia is presented in this report. METHODS: We tracked epidemiological and laboratory findings in a patient with SARS-CoV-2 and Plasmodium falciparum coinfection. A complete blood count was performed using the Sysmex XN-2000 analyser (Sysmex Corporation, Kobe, Japan), coagulation analyses were performed using the BCS XP coagulometer (Siemens Healthineers AG, Erlangen, Germany). Procalcitonin (PCT) and Interleukin-6 (IL-6) were measured by electrochemiluminescence immunoassay (ECLIA) using the Cobas e411 (Roche Diagnostics GmbH, Mannheim, Germany) analyser and high sensitivity troponin I (hsTnI) was measured using the Dimension EXL with LM analyser (Siemens Healthcare Diagnostics, Newark, USA). All other biochemistry analyses were performed using the Olympus AU680 (Beckman Coulter, Brea, California, USA) analyser. White blood cell differential analysis has been performed by examining the blood smear using the CellaVision DM1200 (CellaVision AB, Lund, Sweden) automatic analyser. RESULTS: Even though the patient's initial health condition was disturbed, as a result of the physician's comprehensive anamnesis accompanied by laboratory findings, prompt diagnosis and appropriate therapy were assured, and consequently, the patient recovered. CONCLUSION: In a pandemic, testing each febrile patient for the SARS-CoV-2 virus is of essential importance. However, the possibility of coinfection with another infectious disease agent cannot be disregarded.

Endocannabinoids, Anandamide and 2-Arachidonoylglycerol, as Prognostic Markers of Sepsis Outcome and Complications.

Background: One of the major challenges in improving sepsis care is early prediction of sepsis complications. The endocannabinoid system has been intensely studied in recent years; however, little is known about its role in sepsis in humans. This study aimed to assess the prognostic role of endocannabinoids, anandamide (AEA) and 2-arachidonoylglycerol (2-AG), as early predictors of mortality, invasive mechanical ventilation (IMV) requirement, and length of stay (LOS) in patients with sepsis. Materials and Methods: In total, 106 patients with confirmed sepsis were enrolled in this study. The patients were divided into groups according to mortality outcome (survival, N=53; nonsurvival, N=53), IMV requirement (IMV group, N=26; non-IMV group, N=80), and LOS (LOS <10 days, N=59; LOS ≥10 days, N=47). Patients' clinical status was assessed along with laboratory biomarkers as well as AEA and 2-AG concentration measurements early on admission to emergency units. AEA and 2-AG levels were measured by enzyme-linked immunosorbent assay (ELISA) using an ELISA processor, EtiMax 3000 (DiaSorin, Saluggia, Italy). The predictive value of AEA and 2-AG for the studied sepsis outcomes and complications was analyzed using univariate and multivariate analyses and receiver operating characteristic (ROC) curve analysis. Results: Two endocannabinoids showed no significant difference between survivors and nonsurvivors, although an AEA concentration <7.16 µg/L predicted mortality outcome with a sensitivity of 57% (95% confidence interval [CI] 42-71) and specificity of 80% (95% CI 66-91). AEA concentrations ≤17.84 µg/L predicted LOS ≥10 days with sensitivity of 98% (95% CI 89-100) and specificity of 34% (95% CI 22-47). When analyzing IMV requirement, levels of AEA and 2-AG were significantly lower within the IMV group compared with the non-IMV group (5.94 µg/L [2.04-9.44] and 6.70 µg/L [3.50-27.04], p=0.043, and 5.68 µg/L [2.30-8.60] and 9.58 µg/L [4.83-40.05], p=0.002, respectively). The 2-AG showed the best performance for IMV requirement prediction, with both sensitivity and specificity of 69% (p<0.001). Endocannabinoid AEA was an independent risk factor of LOS ≥10 days (odds ratio [OR] 23.59; 95% CI 3.03-183.83; p=0.003) and IMV requirement in sepsis (OR 0.79; 95% CI, 0.67-0.93; p=0.004). Conclusion: Low AEA concentration is a prognostic factor of hospital LOS longer than 10 days. Lower AEA and 2-AG concentrations obtained at the time of admission to the hospital are predictors of IMV requirement.

Statistical learning confirms the diagnostic significance of the anemia panel in breast cancer.

BACKGROUND: Diagnostic value of available tumor markers, such as cancer antigen CA 15-3 and carcinoembryonic antigen (CEA) in breast cancer is limited. There is an ongoing search for additional, potentially better diagnostic blood markers with improved clinical utility. The aim of this study is to evaluate performance of the approach based on routine blood tests accompanied by a statistical learning tool to the diagnosis of breast cancer. METHODS: Blood was collected from total of 104 subjects which were divided into two groups: breast cancer patients and a control group that consisted of asymptomatic volunteers and patients who had benign breast lesions at the time of blood collection. Random forest statistical learning method and the external method validation have been applied to evaluate diagnostic performance of 31 routine blood tests. RESULTS: The applied statistical learning approach assigned the highest diagnostic importance to the anemia panel among all analyzed blood tests that also included CA 15-3. External validation has shown utility of selected statistical approach - we were able to select tests that provide a diagnostic accuracy comparable to some diagnostic tools described in literature and based on more demanding laboratory techniques, such as gene expression microarrays. CONCLUSIONS: Inclusion of tests for anemia significantly improves diagnostic accuracy for the breast cancer in comparison to the diagnostic accuracy of the CA 15-3 alone. Application of the random forests also enables the reduction of number of laboratory tests needed for the establishment of diagnosis. Differences in relevant test values between the cancer and control group are small but application of multiparametric statistical learning ensured diagnostic accuracy of 72.0% associated by a sensitivity of 64.7% and specificity of 84.9%.

Analytical bias of automated immunoassays for six serum steroid hormones assessed by LC-MS/MS.

INTRODUCTION: There is a growing amount of evidence showing the significant analytical bias of steroid hormone immunoassays, but large number of available immunoassays makes conduction of a single comprehensive study of this issue hardly feasible. Aim of this study was to assess the analytical bias of six heterogeneous immunoassays for serum aldosterone, cortisol, dehydroepiandrosterone sulphate (DHEAS), testosterone, 17-hydroxyprogesterone (OHP) and progesterone using the liquid chromatography coupled to the tandem mass spectrometry (LC-MS/MS). MATERIALS AND METHODS: This method comparison study included 49 serum samples. Testosterone, DHEAS, progesterone and cortisol immunoassays were performed on the Abbott Architect i2000SR or Alinity i analysers (Abbott Diagnostics, Chicago, USA). DiaSorin's Liaison (DiaSorin, Saluggia, Italy) and DIAsource's ETI-Max 3000 analysers (DIAsource ImmunoAssays, Louvain-La-Neuve, Belgium) were chosen for aldosterone and OHP immunoassay testing, respectively. All immunoassays were evaluated against the LC-MS/MS assay relying on the commercial kit (Chromsystems, Gräfelfing, Germany) and LCMS-8050 analyser (Shimadzu, Kyoto, Japan). Analytical biases were calculated and method comparison was conducted using weighted Deming regression analysis. RESULTS: Depending on the analyte and specific immunoassay, mean relative biases ranged from -31 to + 137%. Except for the cortisol, immunoassays were positively biased. For none of the selected steroids slope and intercept 95% confidence intervals simultaneously contained 0 and 1, respectively. CONCLUSIONS: Evaluated immunoassays failed to satisfy requirements for methods' comparability and produced significant analytical biases in respect to the LC-MS/MS assay, especially at low concentrations.

Test results comparison and sample stability study: is the BD Barricor tube a suitable replacement for the BD RST tube?

INTRODUCTION: The aim was to evaluate the BD Barricor tubes by comparison with the BD Rapid Serum Tubes (RST) through measuring 25 analytes and monitoring sample stability after 24 hours and 7 days. MATERIALS AND METHODS: Samples of 52 patients from different hospital departments were examined. Blood was collected in BD RST and BD Barricor tubes (Becton, Dickinson and Company, Franklin Lakes, USA). Analytes were measured by Beckman Coulter AU 480 (Beckman Coulter, Brea, USA), Dimension EXL (Siemens Healthcare Diagnostics, Newark, USA) and ARCHITECT i2000SR (Abbott Diagnostics, Lake Forest, USA). Between-tube comparison for each analyte was performed, along with testing analyte stability after storing samples at 4 °C. RESULTS: BD Barricor tubes showed unacceptable bias compared to BD RST tubes for potassium (K) (- 4.5%) and total protein (4.4%). Analyte stability after 24 hours was acceptable in both tested tubes for most of analytes, except for glucose, aspartate aminotransferase (AST) and lactate dehydrogenase (LD) in BD Barricor and free triiodothyronine in BD RST sample tubes. Analyte stability after 7 days was unacceptable for sodium, K, calcium, creatine kinase isoenzyme MB, AST, LD and troponin I in both samples; additionally for glucose, alkaline phosphatase and albumin in BD Barricor. CONCLUSION: All analytes, except K and total protein, can be measured interchangeably in BD RST and BD Barricor tubes, applying the same reference intervals. For most of the analytes, sample re-analysis can be performed in both tubes after 24 hours and 7 days, although BD RST tubes show better 7-day analytes stability over BD Barricor tubes.

The soluble fms-like tyrosin kinase-1 (sFLT-1) to placental growth factor (PIGF) ratio as a possible indicator for the severity of preeclampsia - single institution experience.

Aim To investigate a potential of the clinical use of the soluble fms-like tyrosine kinase 1 (sFLT-1) to placental growth factor (PlGF) ratio from the perspective of a small hospital centre. Methods Maternal serum samples were analysed at 241/7-28 0/7, and 281/7-320/7 weeks of gestation. The level of sFLT-1 and PIGF was determined by immunoassay platform and used to calculate the sFLT-1/PIGF ratio in 35 pregnant women, and divided into subgroups according to preeclampsia occurrence at the time of delivery: preterm (≤37 weeks) or term (37-42 weeks'), and matched a control group. Results Patients in the preterm delivery group had a significantly higher incidence of intrauterine growth restriction, lower gestational age at the time of delivery, and lower infant birth weight compared to the other two groups. There was a negative correlation between the sFLT-1/PlGF ratio and GA and between the sFLT-1/ PlGF ratio and birth weight at the time of delivery. The value of the sFLT-1/PlGF ratio was significantly higher in the preterm delivery PE group. All the PE group pregnancies ended with caesarean delivery compared to 25% in the control group. However, none of the patients from the PE group had any of the possible complications of preeclampsia nor did they require additional therapy such as blood transfusion or additional non-standard hypertensive therapy. Conclusion The sFLT-1/PlGF ratio could be used as an indicator for the development and estimation of the severity of PE to provide objective evidence for the management of preeclampsia patients, and as a predictive marker of preeclampsia at low cost.

Influence of postoperative analgesia on systemic inflammatory response and postoperative cognitive dysfunction after femoral fractures surgery: a randomized controlled trial.

BACKGROUND AND OBJECTIVES: To investigate the possible effect of postoperatively applied analgesics-epidurally applied levobupivacaine or intravenously applied morphine-on systemic inflammatory response and plasma concentration of interleukin (IL)-6 and to determine whether the intensity of inflammatory response is related to postoperative cognitive dysfunction (POCD). METHODS: This is a randomized, prospective, controlled study in an academic hospital. Patients were 65 years and older scheduled for femoral fracture fixation from July 2016 to September 2017. Inflammatory response was assessed by leukocytes, neutrophils, C reactive protein (CRP) and fibrinogen levels in four blood samples (before anesthesia, 24 hours, 72 hours and 120 hours postoperatively) and IL-6 concentration from three blood samples (before anesthesia, 24 hours and 72 hours postoperatively). Cognitive function was assessed using the Mini-Mental State Examination preoperatively, from the first to the fifth postoperative day and on the day of discharge. RESULTS: The study population included 70 patients, 35 in each group. The incidence of POCD was significantly lower in the levobupivacaine group (9%) than in the morphine group (31%) (p=0.03). CRP was significantly lower in the levobupivacaine group 72 hours (p=0.03) and 120 hours (p=0.04) after surgery. IL-6 values were significantly lower in the levobupivacaine group 72 hours after surgery (p=0.02). The only predictor of POCD in all patients was the level of IL-6 72 hours after surgery (p=0.03). CONCLUSIONS: There is a statistically significant association between use of epidural levobupivacaine and a reduction in some inflammatory markers. Postoperative patient-controlled epidural analgesia reduces the incidence of POCD compared with intravenous morphine analgesia in the studied population. TRIAL REGISTRATION NUMBER: NCT02848599.