(1) Background: “Kidney Disease: Improving Global Outcomes” (KDIGO) provides guidelines for identifying the stages of acute kidney injury (AKI) and chronic kidney disease (CKD). A data-driven rule-based engine was developed to determine KDIGO staging compared to KD-related keywords in discharge letters. (2) Methods: To assess potential differences in outcomes, we compare the patient subgroups with exact KDIGO staging to imprecise or missing staging for all-cause mortality, in-hospital mortality, selection bias and costs by applying Kaplan–Meier analysis and the Cox proportional hazards regression model. We analysed 63,105 in-patient cases from 2016 to 2023 at a tertiary hospital with AKI, CKD and acute-on-chronic KD. (3) Results: Imprecise and missing CKD staging were associated with an 85% higher risk of all-cause and in-hospital mortality (CI: 1.7 to 2.0 and 1.66 to 2.03, respectively) compared to exact staging for any given disease status; imprecise or missing AKI staging increased in-hospital mortality risk by 56% and 57% (CI: 1.43 to 1.70 and 1.37 to 1.81, respectively) in patients with AKI. (4) Conclusions: Exact staging is associated with better outcomes in KD management. Our study provides valuable insight into potential quality and outcome improvements and lower costs, considering elderly patients, women and patients with acute-on-chronic KD as the most vulnerable.

Supplementary Data from Serum Peptidome Profiling Revealed Platelet Factor 4 as a Potential Discriminating Peptide Associated with Pancreatic Cancer

Background The criteria for the diagnosis of kidney disease outlined in the Kidney Disease: Improving Global Outcomes guidelines are based on a patient’s current, historical, and baseline data. The diagnosis of acute kidney injury, chronic kidney disease, and acute-on-chronic kidney disease requires previous measurements of creatinine, back-calculation, and the interpretation of several laboratory values over a certain period. Diagnoses may be hindered by unclear definitions of the individual creatinine baseline and rough ranges of normal values that are set without adjusting for age, ethnicity, comorbidities, and treatment. The classification of correct diagnoses and sufficient staging improves coding, data quality, reimbursement, the choice of therapeutic approach, and a patient’s outcome. Objective In this study, we aim to apply a data-driven approach to assign diagnoses of acute, chronic, and acute-on-chronic kidney diseases with the help of a complex rule engine. Methods Real-time and retrospective data from the hospital’s clinical data warehouse of inpatient and outpatient cases treated between 2014 and 2019 were used. Delta serum creatinine, baseline values, and admission and discharge data were analyzed. A Kidney Disease: Improving Global Outcomes–based SQL algorithm applied specific diagnosis-based International Classification of Diseases (ICD) codes to inpatient stays. Text mining on discharge documentation was also conducted to measure the effects on diagnosis. Results We show that this approach yielded an increased number of diagnoses (4491 cases in 2014 vs 11,124 cases of ICD-coded kidney disease and injury in 2019) and higher precision in documentation and coding. The percentage of unspecific ICD N19-coded diagnoses of N19 codes generated dropped from 19.71% (1544/7833) in 2016 to 4.38% (416/9501) in 2019. The percentage of specific ICD N18-coded diagnoses of N19 codes generated increased from 50.1% (3924/7833) in 2016 to 62.04% (5894/9501) in 2019. Conclusions Our data-driven method supports the process and reliability of diagnosis and staging and improves the quality of documentation and data. Measuring patient outcomes will be the next step in this project.

Background: Serological tests are a powerful tool in the monitoring of infectious diseases and the detection of host immunity. However, manufacturers often provide diagnostic accuracy data generated through biased studies and the performance in clinical practice is essentially unclear. Objectives: We aimed to determine the diagnostic accuracy of various serological testing strategies for (a) identification of patients with previous coronavirus disease-2019 (COVID-19) and (b) prediction of neutralizing antibodies against SARS-CoV-2 in real-life clinical settings. Methods: We prospectively included 2’573 consecutive health-care workers and 1’085 inpatients with suspected or possible previous COVID-19 at a Swiss University Hospital. Various serological immunoassays based on different analytical techniques (enzyme-linked immunosorbent assays, ELISA; chemiluminescence immunoassay, CLIA; electrochemiluminescence immunoassay, ECLIA; lateral-flow immunoassay, LFI), epitopes of SARS-CoV-2 (nucleocapsid, N; receptor-binding domain, RBD; extended RBD, RBD+; S1 or S2 domain of the spike [S] protein, S1/S2), and antibody subtypes (IgG, pan-Ig) were conducted. A positive real-time PCR test from a nasopharyngeal swab was defined as previous COVID-19. Neutralization assays with live SARS-CoV-2 were performed in a subgroup of patients to assess neutralization activity (n=201). Results: The sensitivity to detect patients with previous COVID-19 was ≥85% in anti-N ECLIA (86.8%) and anti-S1 ELISA (86.2%). Sensitivity was 84.7% in anti-S1/S2 CLIA, 84.0% in anti-RBD+ LFI, 81.0% in anti-N CLIA, 79.2% in anti-RBD ELISA, and 65.6% in anti-N ELISA. The specificity was 98.4% in anti-N ECLIA, 98.3% in anti-N CLIA, 98.2% in anti-S1 ELISA, 97.7% in anti-N ELISA, 97.6% in anti-S1/S2 CLIA, 97.2% in anti-RBD ELISA, and 96.1% in anti-RBD+ LFI. The sensitivity to detect neutralizing antibodies was ≥85% in anti-S1 ELISA (92.7%), anti-N ECLIA (91.7%), anti-S1/S2 CLIA (90.3%), anti-RBD+ LFI (87.9%), and anti-RBD ELISA (85.8%). Sensitivity was 84.1% in anti-N CLIA, and 66.2% in anti-N ELISA. The specificity was ≥97% in anti-N CLIA (100%), anti-S1/S2 CLIA (97.7%), and anti-RBD+ LFI (97.9%). Specificity was 95.9% in anti-RBD ELISA, 93.0% in anti-N ECLIA, 92% in anti-S1 ELISA, and 65.3% in anti-N ELISA. Diagnostic accuracy measures were consistent among subgroups. Conclusions: The diagnostic accuracy of serological tests for SARS-CoV-2 antibodies varied remarkably in clinical practice, and the sensitivity to identify patients with previous COVID-19 deviated substantially from the manufacturer’s specifications. The data presented here should be considered when using such tests to estimate the infection burden within a specific population and determine the likelihood of protection against re-infection.

BACKGROUND The criteria for the diagnosis of kidney disease outlined in “The Kidney Disease: Improving Global Outcomes (KDIGO)” are based on a patient’s current, historical and baseline data. The diagnosis of acute (AKI), chronic (CKD) and acute-on-chronic kidney disease requires past measurements of creatinine and back-calculation and the interpretation of several laboratory values over a certain period. Diagnosis may be hindered by unclear definition of the individual creatinine baseline and rough ranges of norm values set without adjustment for age, ethnicity, comorbidities and treatment. Classification of the correct diagnosis and the sufficient staging improves coding, data quality, reimbursement, the choice of therapeutic approach and the patient’s outcome. OBJECTIVE With the help of a complex rule-engine a data-driven approach to assign the diagnoses acute, chronic and acute-on-chronic kidney disease is applied. METHODS Real-time and retrospective data from the hospital’s Clinical Data Warehouse of in- and outpatient cases treated between 2014 – 2019 is used. Delta serum creatinine, baseline values and admission and discharge data are analyzed. A KDIGO based standard query language (SQL) algorithm applies specific diagnosis (ICD) codes to inpatient stays. To measure the effect on diagnosis, Text Mining on discharge documentation is conducted. RESULTS We show that this approach yields an increased number of diagnoses as well as higher precision in documentation and coding (unspecific diagnosis ICD N19* coded in % of N19 generated 17.8 in 2016, 3.3 in 2019). CONCLUSIONS Our data-driven method supports the process and reliability of diagnosis and staging and improves the quality of documentation and data. Measuring patients’ outcome will be the next step of the project.

CONTEXT:α-klotho is an integral membrane protein that serves as a coreceptor for fibroblast growth factor 23 (FGF23) in conjunction with cognate fibroblast growth factor receptors. Proteolytic cleavage sheds the ectodomain of α-klotho (soluble α-klotho) as an endocrine substance into blood, urine, and cerebrospinal fluid.OBJECTIVE:To study the relationship of soluble α-klotho to mineral metabolism in the general population with mainly preserved kidney function.DESIGN:Cross-sectional analysis of the associations between soluble α-klotho with laboratory markers of markers of mineral metabolism in a population-based cohort.SETTING:Three centers in Switzerland including 1128 participants.MEASURES:Soluble full-length α-klotho levels by a specific immunoassay and markers of mineral metabolism.RESULTS:The median serum level of soluble α-klotho was 15.0 pmol/L. Multivariable analyses using α-klotho as the outcome variable revealed a sex-by-PTH interaction: In men, PTH was positively associated with α-klotho levels, whereas this association was negative in women. Plasma phosphate associated with soluble α-klotho levels in an age-dependent manner, changing from a positive association in young adults gradually to a negative association in the elderly. The decline of 1,25 (OH)2 vitamin D3 levels in parallel to the gradual impairment of kidney function was greatly attenuated in the setting of high circulating soluble α-klotho levels.CONCLUSIONS:Soluble α-klotho level is associated with plasma phosphate in an age-dependent manner and with PTH in a sex-dependent manner. Furthermore, our data reveal soluble α-klotho as a modulator of 1,25 (OH)2 vitamin D3 levels in individuals with preserved renal function.

Introduction: Measurements from frozen sample collections are important key indicators in clinical studies. It is a prime concern of biobanks and laboratories to minimize preanalytical bias and variance through standardization. In this study, we aimed at assessing the effects of different freezing and thawing conditions on the reproducibility of medical routine parameters from frozen samples. Materials and Methods: In total, 12 pooled samples were generated from leftover lithium heparinized plasma samples from clinical routine testing. Aliquots of the pools were frozen using three freezing methods (in carton box at -80 degrees C, flash freezing in liquid nitrogen, and controlled-rate freezing [CRF]) and stored at -80 degrees C. After 3 days, samples were thawed using two methods (30 minutes at room temperature or water bath at 25 degrees C for 3 minutes). Ten clinical chemistry laboratory parameters were measured before (baseline) and after freeze-thaw treatment: total calcium, potassium, sodium, alanine aminotransferase, lactate dehydrogenase (LDH), lipase, uric acid, albumin, c-reactive protein (CRP), and total protein. We evaluated the influence of the different preanalytical treatments on the test results and compared each condition with nonfrozen baseline measurements. Results: We found no significant differences between freezing methods for all tested parameters. Only LDH was significantly affected by thawing with fast-rate thawing being closer to baseline than slow-rate thawing. Potassium, LDH, lipase, uric acid, albumin, and CRP values were significantly changed after freezing and thawing compared with unfrozen samples. The least prominent changes compared with unfrozen baseline measurements were obtained when a CRF protocol of the local biobank and fast thawing was applied. However, the observed changes between baseline and frozen samples were smaller than the measurement uncertainty for 9 of the 10 parameters. Discussion: Changes introduced through freezing-thawing were small and not of clinical importance. A slight statistically based preference toward results from slow CRF and fast thawing of plasma being closest to unfrozen samples could be supported.

BACKGROUND:Serological immunoassays that can identify protective immunity against SARS-CoV-2 are needed to adapt quarantine measures, assess vaccination responses, and evaluate donor plasma. To date, however, the utility of such immunoassays remains unclear. In a mixed-design evaluation study, we compared the diagnostic accuracy of serological immunoassays that are based on various SARS-CoV-2 proteins and assessed the neutralizing activity of antibodies in patient sera.METHODS:Consecutive patients admitted with confirmed SARS-CoV-2 infection were prospectively followed alongside medical staff and biobank samples from winter 2018/2019. An in-house enzyme-linked immunosorbent assay utilizing recombinant receptor-binding domain (RBD) of the SARS-CoV-2 spike protein was developed and compared to three commercially available enzyme-linked immunosorbent assays (ELISAs) targeting the nucleoprotein (N), the S1 domain of the spike protein (S1), and a lateral flow immunoassay (LFI) based on full-length spike protein. Neutralization assays with live SARS-CoV-2 were performed.RESULTS:One thousand four hundred and seventy-seven individuals were included comprising 112 SARS-CoV-2 positives (defined as a positive real-time PCR result; prevalence 7.6%). IgG seroconversion occurred between day 0 and day 21. While the ELISAs showed sensitivities of 88.4% for RBD, 89.3% for S1, and 72.9% for N protein, the specificity was above 94% for all tests. Out of 54 SARS-CoV-2 positive individuals, 96.3% showed full neutralization of live SARS-CoV-2 at serum dilutions ≥ 1:16, while none of the 6 SARS-CoV-2-negative sera revealed neutralizing activity.CONCLUSIONS:ELISAs targeting RBD and S1 protein of SARS-CoV-2 are promising immunoassays which shall be further evaluated in studies verifying diagnostic accuracy and protective immunity against SARS-CoV-2.

Background: Significant hope and expectation has been placed on serological immunoassays to test for previous COVID-19 infection, to determine immunity against SARS-CoV2 and to assess the response to vaccination. To date, however the diagnostic utility of such immunoassays remains unclear. In a mixed-design evaluation study, we compared the performance of four serological immunoassays targeting various proteins of the SARS-CoV-2 virus: (a) the receptor binding domain (RBD; ELISA, IgG, IgM), (b) the nucleocapsid (N; ELISA, IgG, IgM), (c) the S1 domain of the spike protein (S1; ELISA, IgG) and (d) the spike protein (S; lateral flow immunoassay, IgG, IgM). Methods: Patients admitted between March and April 2020 to Inselspital University Hospital with confirmed COVID-19 infection were prospectively followed alongside medical staff in a cross-sectional design. Biobank samples from the previous winter period 2018/2019 were additionally tested. An in-house enzyme-linked immunosorbent assay utilising recombinant RBD from the SARS-CoV-2 spike protein was developed and compared to three commercially available serological assays. The primary reference standard was defined as a positive RT-PCR (nasopharyngeal swab). Findings: One-hundred and fifty-nine individuals were included, comprising 25 patients (with 187 data points), 102 medical staff, and 32 biobank samples. Prevalence was 30.8% (n=49). Time to IgG seroconversion occurred between day 2 and day 21 after the start of symptoms and between day 0 and 21 after a positive RT-PCR. Regarding IgG, the area under the receiver operating characteristic curve was 0.97 for RBD (95% confidence interval 0.94 to 1.00), 0.97 for S1 (0.95, 1.00), and 0.94 for N ELISA (0.88, 0.99). Corresponding sensitivity was 79.6% (RBD; 66.4, 88.5), 87.8% (S1; 75.8, 94.3), 67.4% (95% CI 52.5, 80.1), and 71.4% (S LFI; 58.7, 82.1). Specificity was 98.2% (RBD; 93.6, 99.7), 99.1 (S1; 95.0, 100.0), 95.5% (N ELISA 89.7, 98.5), and 95.8% (S LFI). Interpretation: Diagnostic performance of serological immunoassays targeting RBD, S1, N and S appears be adequate for identification of patients with previous COVID-19 infection. This observation should be confirmed by full diagnostic accuracy studies to also derive the precise time-point of seroconversion and presence and duration of immunity. Funding Statement: No specific funding was obtained for the purpose of this study.Declaration of Interests: All authors declare that there is no conflict of interests.Ethics Approval Statement: The study protocol was approved by the appropriate ethics committee and the authorities of the University Hospital and conducted in accordance with the Declaration of Helsinki.

INTRODUCTION:Non-targeted metabolic profiling using high-resolution mass spectrometry (HRMS) is a standard approach for pathway identification despite technical limitations.OBJECTIVES:To assess the performance of combining targeted quadrupole (QQQ) analysis with HRMS for in-depth pathway profiling.METHODS:Serum of exercising patients with type 1 diabetes (T1D) was profiled using targeted and non-targeted assays.RESULTS:Non-targeted analysis yielded a broad unbiased metabolic profile, targeted analysis increased coverage of purine metabolism (twofold) and TCA cycle (three metabolites).CONCLUSION:Our screening strategy combined the benefits of the unbiased full-scan HRMS acquisition with the deeper insight into specific pathways by large-scale QQQ analysis.

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Fibroblast growth factor 23 (FGF23) regulates phosphate homeostasis, and its early rise in patients with chronic kidney disease is independently associated with all-cause mortality. Since inflammation is characteristic of chronic kidney disease and associates with increased plasma FGF23 we examined whether inflammation directly stimulates FGF23. In a population-based cohort, plasma tumor necrosis factor (TNF) was the only inflammatory cytokine that independently and positively correlated with plasma FGF23. Mouse models of chronic kidney disease showed signs of renal inflammation, renal FGF23 expression and elevated systemic FGF23 levels. Renal FGF23 expression coincided with expression of the orphan nuclear receptor Nurr1 regulating FGF23 in other organs. Antibody-mediated neutralization of TNF normalized plasma FGF23 and suppressed ectopic renal Fgf23 expression. Conversely, TNF administration to control mice increased plasma FGF23 without altering plasma phosphate. Moreover, in Il10-deficient mice with inflammatory bowel disease and normal kidney function, plasma FGF23 was elevated and normalized upon TNF neutralization. Thus, the inflammatory cytokine TNF contributes to elevated systemic FGF23 levels and also triggers ectopic renal Fgf23 expression in animal models of chronic kidney disease.

BACKGROUND:Donation after circulatory death (DCD) could significantly improve cardiac graft availability. However, DCD hearts undergo potentially deleterious warm ischemia/reperfusion (I/R). As endothelial damage is a key factor in cardiac I/R injury, we aimed to investigate the tolerance of cardiac and endothelial function after various durations of warm ischemia to improve the timing and choice of cardioprotective therapies. METHODS:Isolated, working rat hearts were perfused for 20 minutes aerobically, then underwent various periods of warm global ischemia and either 30 or 60 minutes of reperfusion. RESULTS:Compared with non-ischemic hearts, recovery of left ventricular work (heart rate-developed pressure product) was significantly reduced at 60 minutes of reperfusion with ≥27 minutes of ischemia (p <0.05 for all), but was unchanged after 21 or 24 minutes of ischemia. Markers of cell death and edema significantly increased with ≥27-minute ischemia compared with non-ischemic hearts (p <0.05 for all). Endothelial-dependent vasodilation was significantly impaired compared with non-ischemic hearts with ≥24 minutes of ischemia, whereas endothelial-independent vasodilation was impaired with ≥27 minutes of ischemia (p <0.05 for all). Furthermore, with ≥24 minutes of ischemia, superoxide production by nitric oxide synthase and peroxynitrite levels were significantly increased compared with non-ischemic hearts, suggesting endothelial nitric oxide synthase (eNOS) uncoupling (p <0.05 for both). CONCLUSIONS:The first signs of endothelial dysfunction after cardiac ischemia occur with less ischemia than cardiac functional alterations, and may result from increased eNOS uncoupling. Strategies aimed at improving eNOS coupling may thus help to optimize both endothelial and myocardial recovery, ultimately facilitating DCD heart transplantation.

Background Thrombin generation (TG) assays evaluate the balance between pro- and anticoagulant forces, to better assess bleeding and thrombotic risks. Although TG readouts obtained with the calibrated automated TG have been investigated in multiple clinical conditions, TG still needs standardization and clinical validation. The automated TG instrument ST Genesia (R) (STG, Stago, Asnieres-sur-Seine, France) provides a normalization of TG parameters based on a reference plasma aiming to reduce the interlaboratory variability and the variability between different measurement runs. Objectives To evaluate STG in a group of healthy adults. Methods Reference intervals in healthy adults and variability of the new standardized reagents for bleeding (BleedScreen) and thrombophilic (ThromboScreen) conditions were determined using STG. Results TG was measured in platelet-free plasma (PFP) samples of 123 healthy adults. Reference intervals were determined for TG parameters. Intra- and interassay coefficients of variation were calculated on quality controls and PFP samples from healthy adults. Oral contraception (OC) possibly influenced TG parameters, resulting in a higher median and a broader reference interval for peak height and endogenous thrombin potential (ETP) in women aged 20 to 49 years than in all other sex and age categories. Therefore, we propose the following reference interval categories: men, women aged OC, women aged OC, and women aged >= 50 years. Normalization was effective to reduce the interassay variability of quality controls for ETP (BleedScreen assay), and peak height and ETP (ThromboScreen assay without thrombomodulin), but had little impact on PFP sample variability. Conclusion STG appears suitable for accurate measurement of TG in healthy adults.

Anaemia affects quality of life and radiographic outcome in rheumatoid arthritis (RA). In a cross-sectional study with 779 patients, we assessed the prognostic potential of the major haematopoietic regulators, hepcidin and erythropoietin, comparing their serum concentrations with respect to different anaemia types, inflammatory activity, anti-cytokine-specific treatment effects and iron deficiency (ID) indices. The results showed that clinical disease activity was more closely associated with haemoglobin levels than with anti-tumour necrosis factor-alpha or interleukin 6 receptor effects. In ID, hepcidin was suppressed, independently of inflammation. Erythropoietin levels were inappropriately low in relation to the degree of anaemia, but, in contrast to low haemoglobin, not directly associated with joint damage progression. Hepcidin and erythropoietin levels are intimately connected with inflammation and ID. Interventional studies on these important targets are already in progress.

BACKGROUND: Cardioprotection and graft evaluation after ischemia-reperfusion (IR) are essential in facilitating heart transplantation with donation after circulatory death. Given the key role of mitochondria in IR, we aimed to investigate the tolerance of cardiac mitochondria to warm, global ischemia and to determine the predictive value of early reperfusion mitochondria-related parameters for post-ischemic cardiac recovery. METHODS: Isolated, working rat hearts underwent 0, 21, 24, 27, 30, or 33 minutes of warm, global ischemia, followed by 60 minutes of reperfusion. Functional recovery (developed pressure x heart rate) was determined at 60 minutes of reperfusion, whereas mitochondrial integrity was measured at 10 minutes of reperfusion. RESULTS: Functional recovery at 60 minutes of reperfusion decreased with >= 27 minutes of ischemia vs no ischemia (n = 7-8/group; p < 0.01). Cytochrome c, succinate release, and mitochondrial Ca2+ content increased with >= 27 minutes of ischemia vs no ischemia (p < 0.05). Ischemia at >= 21 minutes decreased mitochondrial coupling, adenosine 5'-triphosphate content, mitochondrial Ca2+ retention capacity, and increased oxidative damage vs no ischemia (p < 0.05). Reactive oxygen species (ROS) from reverse electron transfer increased with 21 and 27 minutes of ischemia vs no ischemia and 33 minutes of ischemia (p < 0.05), whereas ROS from forward electron transfer increased only with 33 minutes of ischemia vs no ischemia (p < 0.05). Mitochondrial coupling and adenosine 5'-triphosphate content correlated positively and cytochrome c, succinate, oxidative damage, and mitochondrial Ca2+ content correlated negatively with cardiac functional recovery (p < 0.05). CONCLUSIONS: Mitochondrial dysfunction occurs with shorter periods of ischemia than cardiac dysfunction. Mitochondrial coupling, ROS emission from reverse electron transfer, and calcium retention are particularly sensitive to early reperfusion injury, reflecting potential targets for cardioprotection. Indicators of mitochondrial integrity may be of aid in evaluating suitability of donation after circulatory death grafts for transplantation. (C) 2018 International Society for Heart and Lung Transplantation. All rights reserved.

Objectives: The new immunochemistry cobas e 801 module (Roche Diagnostics) was developed to meet increasing demands on routine laboratories to further improve testing efficiency, while maintaining high quality and reliable data. Design and methods: During a non-interventional multicenter evaluation study, the overall performance, functionality and reliability of the new module was investigated under routine-like conditions. It was tested as a dedicated immunochemistry system at four sites and as a consolidator combined with clinical chemistry a three sites. Results: We report on testing efficiency and analytical performance of the new module. Evaluation of sample workloads with site-specific routine request patterns demonstrated increased speed and almost doubled throughput (maximal 300 tests per h), thus revealing that one cobas e 801 module can replace two cobas e 602 modules while saving up to 44% floor space. Result stability was demonstrated by QC analysis per assay throughout the study. Precision testing over 21 days yielded excellent results within and between labs, and, method comparison performed versus the cobas e 602 module routine results showed high consistency of results for all assays under study. In a practicability assessment related to performance and handling, 99% of graded features ma (44%) or even exceeded (55%) laboratory expectations, with enhanced reagent management and loading during operation being highlighted. Conclusion: By nearly doubling immunochemistry testing efficiency on the same footprint as a cobas e 602 module, the new module has a great potential to further consolidate and enhance laboratory testing while maintaining high quality analytical performance with Roche platforms.

Introduction Although centrifugation is performed in almost every blood sample, recommendations on duration and g-force are heterogeneous and mostly based on expert opinions. In order to unify this step in a fully automated laboratory, we aimed to evaluate different centrifugation settings and their influence on the results of routine clinical chemistry analytes. Materials and methods We collected blood from 41 healthy volunteers into BD Vacutainer PST II-heparin-gel- (LiHepGel), BD Vacutainer SST II-serum-, and BD Vacutainer Barricor heparin-tubes with a mechanical separator (LiHepBar). Tubes were centrifuged at 2000xg for 10 minutes and 3000xg for 7 and 5 minutes, respectively. Subsequently 60 and 21 clinical chemistry analytes were measured in plasma and serum samples, respectively, using a Roche COBAS instrument. Results High sensitive Troponin T, pregnancy-associated plasma protein A, ß human chorionic gonadotropin and rheumatoid factor had to be excluded from statistical evaluation as many of the respective results were below the measuring range. Except of free haemoglobin (fHb) measurements, no analyte result was altered by the use of shorter centrifugation times at higher g-forces. Comparing LiHepBar to LiHepGel tubes at different centrifugation setting, we found higher lactate-dehydrogenase (LD) (P = 0.003 to < 0.001) and lower bicarbonate values (P = 0.049 to 0.008) in the latter. Conclusions Serum and heparin samples may be centrifuged at higher speed (3000xg) for a shorter amount of time (5 minutes) without alteration of the analytes tested in this study. When using LiHepBar tubes for blood collection, a separate LD reference value might be needed.

BackgroundCardiac troponin T (cTnT) is elevated after coronary artery bypass grafting surgery. The aim of this study was to determine the association between cTnT elevations between 6 and 12 hours after coronary artery bypass grafting and in‐hospital outcome. Methods and ResultsWe prospectively studied 1722 patients undergoing isolated coronary artery bypass grafting. We assessed the association between conventional cTnT (749 patients) and high‐sensitivity cTnT (hs‐cTnT; 973 patients) 6 to 12 hours postoperatively with in‐hospital major adverse cardiac or cerebrovascular events (MACCE), a composite of all‐cause death, myocardial infarction, or stroke. The prespecified secondary outcome was a safety composite of MACCE, resuscitation, intensive care unit readmission or admission ≥48 hours, inotrope or vasopressor use ≥24 hours, or new‐onset renal insufficiency. Among patients with a conventional cTnT measurement, 92 experienced a MACCE (12%) and 146 experienced a safety composite event (19%). Likewise, for hs‐cTnT, 114 experienced a MACCE (12%) and 153 experienced a safety composite event (16%). Compared with cTnT ≤200 ng/L, each 200‐ng/L increment in cTnT was associated with a monotonous increase in the odds of MACCE and the safety composite outcome. Conventional and hs‐cTnT demonstrated moderate discrimination for MACCE (areas under the fitted receiver operating characteristics curve, 0.72 and 0.77 for conventional and hs‐cTnT, respectively) and the safety composite outcome (areas under the fitted receiver operating characteristics curve, 0.66 and 0.74 for conventional and hs‐cTnT, respectively) and resulted in improved prognostic performance when added to the EuroSCORE. At a cutoff of 800 ng/L, conventional and hs‐cTnT provided clinically relevant power to rule in MACCE and the safety composite outcome. ConclusionscTnT levels assessed between 6 and 12 hours after coronary artery bypass grafting identify patients at increased risk of MACCE or other complications.

Owing to the introduction of next-generation sequencing (NGS) new challenges for diagnostic algorithms and the interpretation of the results for therapeutic decision making in hemato-oncology have arisen. Recurrent somatic mutations crossing the borders between different hematological entities and solid neoplasms have been detected. In analogy to mutant TP53, the same mutation type may occur in myeloid, B- or T-lymphatic malignancies or solid neoplasms. At the same time, a certain mutation can show different prognostic outcomes in different entities and co-existence of certain mutations may change the prognostic relevance. These insights may spark the investigation of targeted therapies with the same substances across different disease entities. This review article summarizes mutations that can emerge in different hematologic and solid malignancies and summarizes other obstacles in the era of modern molecular diagnostics, such as the phenomenon of “clonal hematopoiesis of indeterminate potential” being difficult to interpret in the individual patient.