Background Vitamin D (VD) deficiency (VDD) has been associated with autoimmune diseases, though its exact role is unclear. Consistent with other autoimmune diseases, VDD has been linked to incidence and severity of primary immune thrombocytopenia (ITP), a bleeding disorder characterized by destruction and impaired production of platelets (PLT), mediated by autoantibodies and autoreactive T cells. Severe bleeding can occur in ITP, though bleeding symptoms tend to be mild relative to PLT counts. This is often attributed to immature, highly active PLT. Interestingly, ITP also exhibits higher risk of thrombosis. Recently, VDD was found to enhance PLT activation via the VDR/AKT pathway. Increased platelet reactivity may underlie the moderate bleeding severity and increased thrombotic risk in ITP. We aimed to assess the impact of VD levels on PLT function and investigate if VDD enhances PLT reactivity in ITP. Methods Adult primary ITP patients from the Vienna ITP biobank were analysed (EC No.1843/2016). The SMOG score assessed bleeding severity. Chemiluminescence immunoassay measured serum 25-OH-VD; levels ≥75nmol/L were sufficient (VDS), <75nmol/L and <50nmol/L were considered moderate VDD (mVDD) and severe VDD (sVDD). PLT function was determined by the expression of activated glycoprotein (GP) IIb/IIIa and P-selectin, measured by flow cytometry, naïve and after stimulation by agonists (5μg Thrombin Receptor Activator Peptide 6 (TRAP-6), 1μM adenosine diphosphate (ADP), 80μM arachidonic acid (AA), 0.04μg/mL collagen related protein (CRP)-XL). PLT reactivity and degranulation were defined as the change in GPIIb/IIIa or P-selectin expression respectively after stimulation. Data was compared across VDD groups (sVDD, mVDD, VDS) with Kruskal-Wallis and Dunn's tests. PLT function results were associated with VD levels, sex, age, disease duration, platelet count, and cumulative SMOG score via multivariable linear regressions. Non-linear VD effects were explored using natural splines (5 degrees of freedom). Disease duration, PLT count, and cumulative SMOG score were log2-transformed. Significance of VD was determined by ANOVA F-test, comparing full and restricted models. Results 79 patients (50 (63%) women; 29 (37%) men) with a median age of 47 years were analysed. 14 (17.7%) had sufficient VD, 32 (40.5%) had mVDD, and 33 (41.7%) had sVDD. No significant differences in median past/current treatment lines (sVDD: 2, mVDD: 2, VDS: 2) or steroid use at time of sample collection (7 (21%) sVDD, 6 (19%) mVDD, 5 (36%) VDS) were observed between groups. PLT counts were lowest in sVDD patients (46x109/L), lower than in VDS (53x109/L) and significantly lower than in mVDD patients (78 x109/L; p=0.012). sVDD had a significantly higher cumulative SMOG score than mVDD (p=0.016), though after adjusting for PLT count in ordinal regression, no relationship between bleeding severity and VD levels was found. Naïve P-selectin and GPIIb/IIIa expression did not differ by VD status. While the increase of P-selectin expression in response to AA was higher in mVDD than sVDD and VDS (8% vs 2% vs 1%; p=0.006), there was no clear pattern of altered PLT response to stimuli by VD status. In multivariable regressions, naïve P-selectin or GPIIb/IIIa expression were not associated with VD levels, sex, age, ITP disease duration, PLT count, or bleeding severity. PLT degranulation and reactivity upon addition of agonists was significantly associated with higher PLT counts. No significant associations between VD levels and PLT function upon stimulation were found, except a non-linear association after CRP-XL addition, determined by increases in P-selectin and GPIIb/IIIa expression, with highest/lowest response at VD levels of 80/40nmol/L respectively. Conclusion In our ITP cohort, we could not confirm an association of PLT function with VD levels or states of VD deficiency. Although differences in bleeding severity were observed between patients stratified by VD status, this did not coincide with clear differences in PLT function. While enhanced PLT reactivity was significantly associated with higher PLT counts, there were no clear relationships with VD levels in multivariable regression. Though an association between VDD and ITP disease severity is plausible, the previous observation of increased platelet reactivity in VD deficiency could not be confirmed in adult primary ITP patients.

Background: The sensitivity of the platelet function analyzer (PFA-100, Dade Behring Inc) was shown to be high for the detection of von Willebrand disease (VWD), but limited for platelet function defects. Objectives: To study the diagnostic utility of PFA-100 in mild-to-moderate bleeding disorders and bleeding disorder of unknown cause (BDUC). Methods: PFA-100 closure times (CTs) were measured with collagen-epinephrine (EPI) and collagen-adenosine diphosphate (ADP) cartridges in 818 patients with mild bleeding disorders from the Vienna Bleeding Biobank. Patients on anticoagulation or antiplatelet therapy or thrombocytopenic patients were not included. Results: Only 2% of the 532 BDUC patients had prolonged CTs in PFA (EPI) and PFA (ADP), and 64% in either PFA (EPI) or PFA (ADP). In total, 34% of BDUC patients did not have prolonged CTs in PFA (EPI) or PFA (ADP). These rates were similar to patients with coagulation factor deficiencies (n = 27). The rate of pathologic CTs was significantly higher in patients with VWD (n = 79) and, although less pronounced, in platelet function defect (n =180). In 15 of 18 (83%) VWD patients with von Willebrand factor (VWF) antigen and/or VWF ristocetin cofactor activity levels <30 IU/dL, the PFA-100 was prolonged in both cartridges. No association of the PFA-100 with the bleeding severity was observed in BDUC patients. However, prolonged CTs were associated with higher age, lower hematocrit, lower VWF antigen or VWF ristocetin cofactor activity levels, lower platelet counts, and higher fibrinogen levels in BDUC patients. Conclusion: We could not confirm a diagnostic utility for the PFA-100 in mild-to-moderate bleeding disorder patients, and specifically BDUC. No association between PFA-100 results and bleeding severity was observed in BDUC patients

Background As the healthcare sector evolves, Artificial Intelligence's (AI's) potential to enhance laboratory medicine is increasingly recognized. However, the adoption rates and attitudes towards AI across European laboratories have not been comprehensively analyzed. This study aims to fill this gap by surveying European laboratory professionals to assess their current use of AI, the digital infrastructure available, and their attitudes towards future implementations.Methods We conducted a methodical survey during October 2023, distributed via EFLM mailing lists. The survey explored six key areas: general characteristics, digital equipment, access to health data, data management, AI advancements, and personal perspectives. We analyzed responses to quantify AI integration and identify barriers to its adoption.Results From 426 initial responses, 195 were considered after excluding incomplete and non-European entries. The findings revealed limited AI engagement, with significant gaps in necessary digital infrastructure and training. Only 25.6 % of laboratories reported ongoing AI projects. Major barriers included inadequate digital tools, restricted access to comprehensive data, and a lack of AI-related skills among personnel. Notably, a substantial interest in AI training was expressed, indicating a demand for educational initiatives.Conclusions Despite the recognized potential of AI to revolutionize laboratory medicine by enhancing diagnostic accuracy and efficiency, European laboratories face substantial challenges. This survey highlights a critical need for strategic investments in educational programs and infrastructure improvements to support AI integration in laboratory medicine across Europe. Future efforts should focus on enhancing data accessibility, upgrading technological tools, and expanding AI training and literacy among professionals. In response, our working group plans to develop and make available online training materials to meet this growing educational demand.

Electronic health records will increasingly aggregate longitudinal laboratory results from multiple providers, but availability alone does not guarantee safe interpretation. We present guidance, developed by laboratory professionals with the DGKL medical informatics division, for cumulative displays that are clinically meaningful. The core principle is to group medically comparable analyses while preserving laboratory provenance so that clinicians can follow true patient trends without conflating them with laboratory-induced variation. Comparability is defined algorithmically from Logical Observation Identifiers Names and Codes (LOINC) axis: analyses estimating the same patient property (allowing serum/plasma system equivalence and mathematically convertible properties such as substance vs. mass concentration) are grouped; coding of units is harmonized via Unified Code for Units of Measure (UCUM) with consistent conversion of numeric results and corresponding reference intervals, including inequality qualifiers. Analyte-specific conversion factors should come from authoritative sources; for poorly standardized measurands (e.g., tumor markers) or when conversions are inappropriate (e.g., Lp(a)), results remain separated. Methodological distinctions that affect interpretation - such as screening vs. confirmatory drug testing and point-of-care testing - are displayed independently to signal potential analytical discontinuities. A standardized, medically meaningful default result sequence - derived from LOINC metadata and clinical nomenclatures, with alphabetic naming as a pragmatic fallback - supports cross-laboratory aggregation; rare or novel tests lacking robust standardization remain as free text. The rules-based approach updates seamlessly with LOINC releases and remains compatible with the Nomenclature for Properties and Units (NPU), facilitating cross-border exchange within the European Health Data Space. While harmonized presentation improves trend analysis, true comparability ultimately requires measurement procedures traceable to reference methods and materials.

Abstract: The G haplotype is a group of co-inherited single nucleotide variants in the F5 gene that reduce venous thromboembolism (VTE) risk. Although 7% of the population is homozygous for the G haplotype (F5-G/G), the underlying mechanism of VTE protection is poorly understood. Using RNA sequencing data from 4651 blood donors in the INTERVAL study, we detected a rare excision event at the factor V (FV)-short splice sites in 5% of F5-G/Gs carriers as compared with 2.16% of homozygotes for the F5 reference sequence (F5-ref; P = .003). Highly elevated (∼10-fold) FV-short, a FV isoform that lacks most of the B-domain, has been linked with increased tissue factor inhibitor α (TFPIα) levels in rare hemorrhagic diathesis, including East Texas bleeding disorder. To ascertain whether the enhanced FV-short splicing seen in F5-G/G INTERVAL participants translated to increased plasma FV-short levels, we analyzed plasma samples from 7 F5-G/G and 13 F5-ref individuals in a recall-by-genotype study. A ∼2.2-fold higher amount of FV-short was found in a plasma pool from F5-G/G participants when compared with the pool of F5-refs (P = .029), but there was no difference in the total FV levels. Although no significant difference in TFPI levels were found, F5-G/Gs showed a ∼1.4-fold TFPI-dependent increase in lag time to thrombin generation than F5-refs (P = .0085). Finally, in an analysis of 117 699 UK Biobank participants, we discovered that, although being protective against VTE, the G haplotype also confers an increase in bleeding episodes (P = .011). Our study provides evidence that the effect of the common G haplotype is mediated by the FV-short/TFPI pathway.

In patients with a mild to moderate bleeding disorder (MBD) and abnormal light transmission aggregometry (LTA), a platelet function defect (PFD) is suspected. However, in many patients with PFD, the underlying mechanism remains elusive. Given the essential role of lipids in platelet signaling, platelet lipid profiles in MBD patients with unexplained PFD may provide valuable diagnostic and mechanistic insights. This prospective cohort study investigated platelet lipidomes in patients with PFD of unknown cause from the Vienna Bleeding Biobank (VIBB). Using a standardized lipidomics workflow, we analyzed platelets from 27 patients and 19 age- and sex-matched controls and found that sex-specific lipid shifts emerged exclusively within the patient cohort, with greater deviations in females. Furthermore, lipid alterations correlated with impaired platelet aggregation and were predictive of responses to ADP and TRAP-6 stimuli in LTA experiments. Baseline and stimulated platelet analyses in a female subgroup showed intrinsic lipidomic changes, including upregulated polyunsaturated triacylglycerols (PUFA-TG), acylcarnitines (CAR), and reduced lysophosphatidylethanolamines (LPE). This study emphasizes lipidomic profiling as a promising diagnostic tool for unexplained platelet dysfunction and highlights TG, CAR, and LPE as potential therapeutic targets. Further research into lipid-driven platelet regulation may advance personalized treatments and improve clinical outcomes for patients with MBD.

BACKGROUND:Autoantibody (AAb)-induced platelet desialylation, resulting in increased hepatic platelet clearance, has previously been reported as pathomechanism in immune thrombocytopenia (ITP). OBJECTIVES:To elucidate platelet desialylation capacity and its impact on clinical outcomes in adult primary ITP patients from the Vienna ITP biobank (EC 1843/2016). METHODS:Desialylation was assessed using a lectin-binding assay which evaluated the exposure of β-galactose and N-acetylglucosamine on platelets. RESULTS:Sera from 133 ITP patients (28% newly diagnosed and 72% persistent/chronic) were investigated, where sera of 38 patients (29%) induced platelet desialylation. Patients with desialylation capacity compared with those without more commonly had platelet-bound AAbs (n = 19/46, 50% vs n = 27/95, 28.4%; P = .018), thrombopoietin levels > 50 pg/mL (n = 23/34, 67.6% vs n = 37/38, 42.0%; P = .011), and higher bleeding scores (median [IQR], 3.0 [1.0-5.0] vs 1.0 [0.0-4.0]; P = .043). While the bleeding score overall, specifically the skin bleeding score, thrombopoietin > 50 pg/mL, and the presence of AAbs showed a significant association in univariate analysis with desialylating AAbs, only positivity for antiplatelet AAbs remained significantly associated in multivariate binary logistic regression analysis. In contrast, no association was seen between platelet desialylation capacity and disease duration, ITP treatment, or previous splenectomy. CONCLUSION:Platelet desialylation capacity was seen in a third of primary ITP patients and was associated with platelet-bound antiglycoprotein IIbIIIa AAbs, although more studies are required to establish the linkage with antiglycoprotein Ib-IX AAbs. Desialylation capacity was associated with a more severe bleeding phenotype, while, in contrast to previous data, an association with platelet counts, disease duration, ITP treatment, and splenectomy refractoriness was not confirmed.

Recent changes in the regulatory assessment of in vitro medical tests reflect a growing recognition of the need for more stringent clinical evidence requirements to protect patient safety and health. Under current regulations in the United States and Europe, when needed for regulatory approval, clinical performance reports must provide clinical evidence tailored to the intended purpose of the test and allow assessment of whether the test will achieve the intended clinical benefit. The quality of evidence must be proportionate to the risk for the patient and/or public health. These requirements now cover both commercial and laboratory developed tests (LDT) and demand a sound understanding of the fundamentals of clinical performance measures and study design to develop and appraise the study plan and interpret the study results. However, there is a lack of harmonized guidance for the laboratory profession, industry, regulatory agencies and notified bodies on how the clinical performance of tests should be measured. The Working Group on Test Evaluation (WG-TE) of the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM) is a multidisciplinary group of laboratory professionals, clinical epidemiologists, health technology assessment experts, and representatives of the in vitro diagnostic (IVD) industry. This guidance paper aims to promote a shared understanding of the principles of clinical performance measures and study design. Measures of classification performance, also referred to as discrimination, such as sensitivity and specificity are firmly established as the primary measures for evaluating the clinical performance for screening and diagnostic tests. We explain these measures are just as relevant for other purposes of testing. We outline the importance of defining the most clinically meaningful classification of disease so the clinical benefits of testing can be explicitly inferred for those correctly classified, and harm for those incorrectly classified. We introduce the key principles and a checklist for formulating the research objective and study design to estimate clinical performance: (1) the purpose of a test e.g. diagnosis, screening, risk stratification, prognosis, prediction of treatment benefit, and corresponding research objective for assessing clinical performance; (2) the target condition for clinically meaningful classification; (3) clinical performance measures to assess whether the test is fit-for-purpose; and (4) study design types. Laboratory professionals, industry, and researchers can use this checklist to help identify relevant published studies and primary datasets, and to liaise with clinicians and methodologists when developing a study plan for evaluating clinical performance, where needed, to apply for regulatory approval.

The EU AI Act came into force on August 1, 2024, and since February 2, 2025, employees must be sufficiently trained in the use of AI systems (Article 4 of the EU AI Act). But what does “sufficient” AI literacy mean in the context of the EU AI Act? This article highlights the statements of the EU AI Act on AI literacy and aims to help in the development of training curricula. Medical laboratories offer numerous areas of application for AI, including improving the organization of laboratory processes and knowledge management. Even though there are no mandatory training requirements yet, basic data literacy is considered a prerequisite for AI Literacy. Three major topics are identified for AI literacy training: a general knowledge on the content of the EU AI Act, fundamentals of artificial intelligence, and context-specific training. This combination of fundamental data expertise and fundamental AI expertise will enable Article 4 of the EU AI Act to be fulfilled “to the best of ability” and lay the foundation for the transformation of medical laboratories.

In the last decades, clinical laboratories have significantly advanced their technological capabilities, through the use of interconnected systems and advanced software. Laboratory Information Systems (LIS), introduced in the 1970s, have transformed into sophisticated information technology (IT) components that integrate with various digital tools, enhancing data retrieval and exchange. However, the current capabilities of LIS are not sufficient to rapidly save the extensive data, generated during the total testing process (TTP), beyond just test results. This opinion paper discusses qualitative types of TTP data, proposing how to divide laboratory-generated information into two categories, namely metadata and peridata. Being both metadata and peridata information derived from the testing process, it is proposed that the first is useful to describe the characteristics of data, while the second is for interpretation of test results. Together with standardizing preanalytical coding, the subdivision of laboratory-generated information into metadata or peridata might enhance ML studies, also by facilitating the adherence of laboratory-derived data to the Findability, Accessibility, Interoperability, and Reusability (FAIR) principles. Finally, integrating metadata and peridata into LIS can improve data usability, support clinical utility, and advance AI model development in healthcare, emphasizing the need for standardized data management practices.

The integration of artificial intelligence (AI) and machine learning (ML) into laboratory medicine shows promise for advancing diagnostic, prognostic, and decision-support tools; however, routine clinical implementation remains limited and heterogeneous. Laboratory data presents unique methodological and semantic complexities - method dependency, analyte-specific variation, and contextual sensitivity-not adequately addressed by general-purpose AI reporting guidelines. To bridge this gap, the EFLM Committee on Digitalisation and Artificial Intelligence (C-AI) proposes an expanded checklist to support assessment of requirements and recommendations for the development of AI/ML models based on laboratory data. Building upon the widely adopted ChAMAI checklist (Checklist for assessment of medical AI), our proposal introduces six additional items, each grounded in the CRoss Industry Standard Process for Data Mining (CRISP-DM) framework and tailored to the specificities of laboratory workflows. These extensions address: (1) explicit documentation of laboratory data characteristics; (2) consideration of biological and analytical variability; (3) the role of metadata and peridata in contextualizing results; (4) analyte harmonization and standardization practices; (5) rigorous external validation with attention to dataset similarity; and (6) the implementation of FAIR data principles for transparency and reproducibility. Together, these recommendations aim to foster robust, interpretable, and generalizable AI systems that are fit for deployment in clinical laboratory settings. By incorporating these laboratory-aware considerations into model development pipelines, researchers and practitioners can enhance both the scientific rigor and practical applicability of AI tools. We advocate for the adoption of this extended checklist by developers, reviewers, and regulators to promote trustworthy and reproducible AI in laboratory medicine.

Background A platelet function defect (PFD) is diagnosed in patients with mild to moderate bleeding disorders and abnormalities in light transmission aggregometry. The influence of platelet aging on platelet function in PFD remains unclear. Objectives We aimed to assess surface receptor profiles of mature and immature platelets in patients with suspected PFD and their relation to hemostatic function. Methods Twenty-two patients with light transmission aggregometry alterations from the Vienna Bleeding Biobank were compared with 19 age- and biological sex-matched healthy controls. Blood samples were immediately analyzed using platelet function analyzer-100 system (Dade Behring Inc) with epinephrine cartridges. Platelet subtypes were characterized via surface receptor profiling of CD9, CD31, CD36, CD40L, CD42b, CD62P, CD63, CD107, and Toll-like receptors (TLRs) 2, 4, and 9, and immature (RNA-rich) platelets were quantified. Results Platelet counts, immature platelet fraction, and mean platelet volume, as well as other hemostatic parameters, did not differ between patients and controls, while patients demonstrated prolonged closure times on platelet function analyzer-100 (Dade Behring Inc). Principal component analysis highlighted distinct glycoprotein expression patterns between immature and mature platelets in both their resting state and after activation in patients and healthy controls. In patients, immature platelets expressed lower levels of CD62P, CD36, CD31, TLR2, and TLR4 but increased TLR9 expression, while mature platelets showed no differences. These distinct surface receptor patterns of immature platelets were partly associated with reduced thrombin receptor activator peptide 6-induced platelet aggregation. Conclusion This study demonstrates significant alterations in surface receptor expression on immature platelets in patients with suspected PFD, underscoring immature platelets as crucial link to platelet biology and clinical symptoms in this patient population.

Rationale: Passenger lymphocyte syndrome (PLS) may complicate minor ABO mismatched lung transplantation (LuTX) via donor-derived red cell antibody-induced hemolysis. Objectives: To ascertain the incidence and specificity of PLS-relevant antibodies among the study population as well as the dynamics of hemolysis parameters and the transfusion requirement of patients with or without PLS. Methods: In this cohort study, 1,011 patients who received LuTX between January 2010 and June 2019 were studied retrospectively. Prospectively, 87 LuTX (July 2019 to June 2021) were analyzed. Postoperative ABO antibody and hemolytic marker determinations, transfusion requirement, and duration of postoperative hospital care were analyzed. Retrospectively, blood group A recipients of O grafts with PLS were compared with those without. Measurements and Main Results: PLS affected 18.18% (retrospective) and 30.77% (prospective) of A recipients receiving O grafts, 5.13% of B recipients of O grafts, and 20% of AB patients receiving O transplants. Anti-A and anti-A1 were the predominant PLS-inducing antibodies, followed by anti-B and anti-A,B. Significantly lower hemoglobin values (median, 7.4 vs. 8.3 g/dl; P = 0.0063) and an approximately twice as high percentage of patients requiring blood transfusions were seen in PLS. No significant differences in other laboratory markers, duration of hospital stay, or other complications after LuTX were registered. Conclusions: Minor ABO incompatible LuTX recipients are at considerable risk of developing clinically significant PLS. Post-transplant monitoring combining red cell serology and hemolysis marker determination appears advisable so as not to overlook hemolytic episodes that necessitate antigen-negative transfusion therapy.

In the last decades, clinical laboratories have significantly advanced their technological capabilities, through the use of interconnected systems and advanced software. Laboratory Information Systems (LIS), introduced in the 1970s, have transformed into sophisticated information technology (IT) components that integrate with various digital tools, enhancing data retrieval and exchange. However, the current capabilities of LIS are not sufficient to rapidly save the extensive data, generated during the total testing process (TTP), beyond just test results. This opinion paper discusses qualitative types of TTP data, proposing how to divide laboratory-generated information into two categories, namely metadata and peridata. Being both metadata and peridata information derived from the testing process, it is proposed that the first is useful to describe the characteristics of data, while the second is for interpretation of test results. Together with standardizing preanalytical coding, the subdivision of laboratory-generated information into metadata or peridata might enhance ML studies, also by facilitating the adherence of laboratory-derived data to the Findability, Accessibility, Interoperability, and Reusability (FAIR) principles. Finally, integrating metadata and peridata into LIS can improve data usability, support clinical utility, and advance AI model development in healthcare, emphasizing the need for standardized data management practices.

Objectives Reference intervals (RI) play a decisive role in the interpretation of medical laboratory results. An important step in the determination of RI is age- and sex specific partitioning, which is usually based on an empirical approach by graphical representation. In this study, we evaluate an automated machine learning approach.Methods This study uses pediatric data from the CALIPER RI (Canadian laboratory initiative on pediatric reference intervals) study. The calculation of potential partitions is carried out using a regression tree model included in the rpart package of the statistical programming language R. The Harris & Boyd method is used to compare the corresponding partitions suggested by rpart and CALIPER. For better comparability, the reference ranges of the partitions of both approaches are then calculated using reflimR.Results Most of the partitions suggested by rpart or CALIPER show sufficient heterogeneity among themselves to justify age- and/or sex-specific RI partitioning. With only few individual exceptions, both methods yield comparable results. The partitions of both approaches for albumin and gamma-glutamyltransferase are very similar to each other. For creatinine rpart suggests a slightly earlier distinction between the sexes. Alkaline phosphatase shows the most pronounced differences. In addition to a considerable earlier sex split, rpart suggests different age intervals for both sexes, resulting in three partitions for females and four partitions for males.Conclusions Our findings indicate that the automated analysis provided by rpart yields results that comparable to traditional methods. Nevertheless, the medical plausibility of the automatic suggestions needs to be validated by human experts.

Small-molecule EGFR inhibitors have distinctly improved the overall survival especially in EGFR-mutated lung cancer. However, their use is often limited by severe adverse effects and rapid resistance development. To overcome these limitations, a hypoxia-activatable Co(III)-based prodrug (KP2334) was recently synthesized releasing the new EGFR inhibitor KP2187 in a highly tumor-specific manner only in hypoxic areas of the tumor. However, the chemical modifications in KP2187 necessary for cobalt chelation could potentially interfere with its EGFR-binding ability. Consequently, in this study, the biological activity and EGFR inhibition potential of KP2187 was compared to clinically approved EGFR inhibitors. In general, the activity as well as EGFR binding (shown in docking studies) was very similar to erlotinib and gefitinib (while other EGFR-inhibitory drugs behaved different) indicating no interference of the chelating moiety with the EGFR binding. Moreover, KP2187 significantly inhibited cancer cell proliferation as well as EGFR pathway activation in vitro and in vivo. Finally, KP2187 proved to be highly synergistic with VEGFR inhibitors such as sunitinib. This indicates that KP2187-releasing hypoxia-activated prodrug systems are promising candidates to overcome the clinically observed enhanced toxicity of EGFR-VEGFR inhibitor combination therapies.

Introduction: Bleeding disorder of unknown cause (BDUC) is a diagnosis of exclusion and patients share a similar phenotype with established mild bleeding disorders (MBDs) such as platelet function defects (PFD), mild von Willebrand disease (VWD), or mild coagulation factor deficiencies (CFD). The integration of global assays, such as the Platelet Function Analyzer (PFA)-100, as screening tools for MBDs lacks universal endorsement and is currently not recommended in clinical routine. While the PFA-100 has demonstrated noteworthy sensitivity in the detection of VWD, data regarding its clinical utility in PFD are conflicting and data on PFA-100 in BDUC patients are scarce. Aims: To evaluate the diagnostic utility of PFA-100 in BDUC compared to other MBDs, and to investigate the influence of various parameters on PFA-100 results. Methods: This study included 821 patients enrolled in the Vienna Bleeding Biobank (VIBB), a single-center cohort study on individuals with non-trivial bleeding tendencies who were referred to our tertiary center until December 2022 (EC 604/2009). Patients under anticoagulation or antiplatelet treatment were excluded. PFA-100® assessments (Dade Behring Inc., Newark, Delaware, USA) were conducted on citric acid anticoagulated whole blood samples obtained at study inclusion using two different cartridges: Dade® PFA-100 Collagen/Epinephrine (EPI) and Dade® PFA-100 Collagen/adenosindiphosphat (ADP; Siemens Healthcare Diagnostics Products GmbH, Marburg, Germany). Results: PFA-100 data of 535 BDUC patients (65%) were compared to those of 79 patients (10%) with VWD, 180 (22%) with PFD, and 27 (3%) with CFD. Clinical characteristics are shown in Table 1. In patients with BDUC, the median (interquartile range) closure time (CT) measured by the PFA-100-EPI assay was 102 seconds (88-117), and 135 seconds (112-163; Table 1) with the PFA-100-ADP assay. The CTs were shorter in BDUC compared to VWD (PFA-100-EPI: p<.001, PFA-100-ADP: <p.001) and PFD (PFA-100-EPI: p<.001, PFA-100-ADP: <p.001), while no significant difference was observed compared to CFD (PFA-100-EPI: p=0.2; PFA-100-ADP: p=0.7). Based on in-house reference ranges, CT was normal in both PFA-100 assays in 180 (34%) of all BDUC patients, while 341 patients (64%) had at least one abnormal CT and 11 patients (2%) had abnormal CTs in both PFA-100 assays. A similar distribution was observed in patients with CFD (no abnormal CT: 33%, 1 abnormal CT: 63%, 2 abnormal CTs: 4%). Statistical analysis revealed a significant difference in the number of pathological CTs between BDUC and VWD (no abnormal CT: 9%, 1 abnormal CT: 58%, 2 abnormal CTs: 33%; p<.001), as well as between BDUC and PFD (no abnormal CT: 24%, 1 abnormal CT: 69%, 2 abnormal CTs: 7%; p=0.003). In respect to bleeding severity, in BDUC patients no correlation was found between CTs in the PFA-100-EPI and PFA-100-ADP assays with the Vicenza bleeding score (PFA-100-EPI Spearman rho (r)=0.06, p=0.06; PFA-100-ADP r=0.02, p=0.6), ISTH-BAT (PFA-100-EPI r= 0.06, p=0.3; PFA-100-ADP r= 0.10, p=0.06), or the number of bleeding manifestations (PFA-100-EPI r=0.01, p=0.81; PFA-100-ADP r=-01, p=0.8). The number of pathological PFA-100 was not associated with specific bleeding symptoms assessed by the Vicenza score in BDUC (data not shown). Using a Tobit regression model, we analyzed the association of clinical and laboratory parameters with prolonged CTs in both PFA-100 assays in BDUC, as shown in Table 2. In multivariable linear regression, reduced hematocrit, lower VWF:Ag/VWF:RCo levels, reduced platelet count and higher fibrinogen levels were associated with longer CTs. Increasing age was additionally associated with a longer CT in PFA-100-ADP. Conclusion: In a minority of BDUC patients CT was prolonged in both PFA-100 (EPI and ADP) assays. PFA-100 was not associated with the bleeding severity or specific symptoms in BDUC. In BDUC patients, lower hematocrit, lower platelet count, lower VWF levels and interestingly higher fibrinogen levels were associated with a prolonged CT. Our descriptive results do not suggest the integration of PFA-100 as screening tool for BDUC patients.

Objectives: Even though most physicians and professionals in laboratory medicine have received basic training in statistics, experience shows that a general understanding of data analysis is not yet available on a broad scale. Therefore, data literacy, data-driven decision making, and computational thinking should be implemented in future educational training. To evaluate the state of digital competence among young scientists (YS) in laboratory medicine, we launched a worldwide online survey. Methods: A global online survey was conducted from 25/05/2022 to 26/06/2022 and was disseminated to YS who are listed in three large networks: YS of the DGKL, the EFLM Task Group-YS, and IFCC Task Force-YS and its corresponding members, covering a base of 53 countries. Results: A total of 119 young scientists from 40 countries participated in this survey. 80 % did not learn digital skills in their academic education but 96 % felt they needed to. Digital literacy was associated with terms such as programming, artificial intelligence and machine learning, statistics, communication, Big Data and data analytics. Conclusions: The results of our survey show that more knowledge and training in the area of digital skills is not just necessary, but also wanted by young scientists. A varied learning environment consisting of tutorial articles, videos, exercises, technical articles, collection of helpful links, online meetings and in person bootcamps is crucial to meet the challenges of an international project with different languages, health systems and time zones.

Objectives: ChatGPT, a tool based on natural language processing (NLP), is on everyone's mind, and several potential applications in healthcare have been already proposed. However, since the ability of this tool to interpret laboratory test results has not yet been tested, the EFLM Working group on Artificial Intelligence (WG-AI) has set itself the task of closing this gap with a systematic approach.Methods: WG-AI members generated 10 simulated laboratory reports of common parameters, which were then passed to ChatGPT for interpretation, according to reference intervals (RI) and units, using an optimized prompt. The results were subsequently evaluated independently by all WG-AI members with respect to relevance, correctness, helpfulness and safety.Results: ChatGPT recognized all laboratory tests, it could detect if they deviated from the RI and gave a test-by-test as well as an overall interpretation. The interpretations were rather superficial, not always correct, and, only in some cases, judged coherently. The magnitude of the deviation from the RI seldom plays a role in the interpretation of laboratory tests, and artificial intelligence (AI) did not make any meaningful suggestion regarding follow-up diagnostics or further procedures in general.Conclusions: ChatGPT in its current form, being not specifically trained on medical data or laboratory data in particular, may only be considered a tool capable of interpreting a laboratory report on a test-by-test basis at best, but not on the interpretation of an overall diagnostic picture. Future generations of similar AIs with medical ground truth training data might surely revolutionize current processes in healthcare, despite this implementation is not ready yet.