The validity, psychometric robustness, and educational effectiveness of the Pharmacology Concept Inventory (PCI), including its ability to identify students’ misconceptions, were assessed across diverse educational settings and student cohorts to evaluate its potential as a concept-based assessment tool in pharmacology education. The PCI comprised 24 multiple-choice items, developed through a triangulated strategy that integrated core pharmacology concepts, expert input, and empirical evidence, and was informed by documented student misconceptions. This multinational validation and effectiveness evaluation included 1211 students from 23 institutions across 12 countries. The analyses examined the instrument’s structure, item- and test-level psychometric properties, and measurement invariance across demographic and educational subgroups. Factor analysis indicated that the PCI primarily measures a single underlying dimension of pharmacological proficiency, with some weak secondary multidimensional groupings of items. The PCI demonstrated consistently strong item discrimination and a moderate level of difficulty, with convergent findings across factor analysis, cognitive diagnostic assessment, and other psychometric approaches. Exploratory cognitive diagnostic estimates showed that mastery varied across concepts, with drug tolerance having the highest probability (68%) and the structural activity relationship the lowest (33%). Year of study and academic program were significant predictors of pharmacological proficiency, with early-year and medical students more likely to achieve mastery than late-year and non-medical students. The PCI also identified a proficiency distribution, providing insight into the extent of variability and the gaps in learners' understanding of core pharmacology. Overall, this study establishes the PCI as a valid, reliable, and diagnostically informative tool for assessing conceptual understanding and supporting evidence-based pharmacology education across international contexts.

BACKGROUND AND AIMS:Nonsteroidal anti-inflammatory drugs (NSAIDs) can cause small intestinal injury, which largely depends on the presence of gut bacteria and the immune responses triggered by them. While previous studies have emphasised the role of Toll-like receptor 4 (TLR4) and TLR2 in enteropathy, the functional significance of TLR5, a receptor for bacterial flagellin, remains elusive. Thus, we aimed to assess the impact of TLR5 activation and inhibition on NSAID enteropathy in mice. MATERIALS AND METHODS:Enteropathy was induced using indomethacin (IND). Mucosal injury, inflammation and the mRNA levels of TLR5, TLR4 and TLR2 were assessed after 24 h. The intestinal flagellin load was measured using Western blot, while bacterial counts were assessed using qPCR. Flagellin (10 and 30 μg) and TH1020 (10 μg) were administered intraperitoneally. TLR5 levels were also determined in the ileum of naproxen-treated rats. KEY FINDINGS:NSAID enteropathy was associated with downregulation of TLR5, intestinal inflammation and apoptosis, mucosal histological damage, higher bacterial counts and flagellin load in the intestine, and upregulation of TLR2 and TLR4. IND-induced changes, except the fall in TLR5 expression, were reduced or completely prevented in animals that had been pretreated with flagellin. Importantly, flagellin induced similar robust protection when administered four hours after IND. In contrast to flagellin, treatment with the potent and selective TLR5 antagonist TH1020 exacerbated the intestinal inflammation caused by IND. SIGNIFICANCE:Our findings reveal that TLR5 activation protects against NSAID-induced enteropathy and may therefore be a new therapeutic avenue.

Abstract The Digital Drug Assignment (DDA) system is a knowledge-graph-based computational method that automates reasoning at the patient level and scores molecularly targeted agents (MTAs) based on the full tumor genomic data. This approach was predictive of relative benefit of the agents as used in the SHIVA01 trial (DOI: 10.1038/s41698-021-00191-2). Here, we evaluated the predictive power of DDA on a larger scale by analyzing tumor genomic and drug sensitivity data from the GDSC database. Our study was based on data from 659 cell lines derived from a broad spectrum of solid tumors. Corresponding drug sensitivity data were available for 34,713 treatment datapoints involving 87 types of MTAs. All tumor genomic profiles were processed using DDA, which scores MTAs and stratifies them according to predicted efficacy. Consequently, the same MTA can receive different drug scores across tumors, depending on their individual molecular profiles. Treatments were then ranked for each tumor based on their DDA scores, resulting in 72 treatment groups defined by ranking positions (i.e., drugs ranked at the same position across tumors formed one treatment group). Sensitivity to treatment was determined using Z-scores of IC50 values, with treatments showing negative Z-scores classified as sensitive. Among the top-ranked MTAs, 54% of treatments were sensitive, with sensitivity gradually decreasing across lower-ranking groups and reaching 0% in the bottom group. A linear trend across DDA score rank groups from top to bottom was confirmed by the Cochran-Armitage trend test (Z = -10.42, p = 2.08e-25), indicating a very strong negative trend across ordered groups from the top towards the bottom. Increased confidence in the benchmark drug response classification was achieved by excluding treatments around the median with progressively larger absolute IC50 Z-score thresholds. With IC50 Z-score exclusion thresholds of absolute 1, 2, 2.5, and 3, the sensitivity of cell lines to top-ranked treatments was 59%, 67%, 74%, and 83%, respectively, while remaining 0% in the bottom groups in all cases. Thus, increasing confidence in drug response correlated with higher predictive accuracy. Although group sizes decreased with stricter thresholds - reducing statistical power - the results remained highly significant throughout, the Cochran-Armitage trend test Z-values gradually increased from -8.33 to -4.68 (all p < 0.001). These results demonstrate the predictive power of DDA-score-based treatment ranking across solid tumors and MTAs, with top-ranked drugs having the greatest efficacy. The findings strongly support the notion that there is a correlation between aggregated scientific evidence and drug sensitivity. DDA can potentially address challenges with complex molecular profiles in routine clinical settings and clinical trial design. Citation Format: Robert Doczi, Akos Takacs, Anna Dirner, Dora Lakatos, Barbara Vodicska, Dora Gorog-Tihanyi, Reka Szalkai-Denes, Eniko Kispeter, Andras Makkos, Aniko Gorbe, Peter Ferdinandy, William T. Beck, Christophe Le Tourneau, Petak Istvan. Molecularly-informed prediction of treatment efficacy in GDSC cell line data using computational reasoning [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 2719.

Small cell lung cancer (SCLC) is a highly aggressive malignancy with poor survival outcomes. The CD70-CD27 axis has been implicated in immune regulation and tumor progression across cancers, but its role in SCLC has not yet been elucidated. This research explores the expression patterns and prognostic significance of CD70 and CD27 in early-stage SCLC. In this retrospective study, we analyzed 190 surgically resected SCLC tumor samples using immunohistochemistry (IHC) for CD70 and CD27 expression and RNAscope for CD70 RNA detection. Immune infiltration was assessed using CD45, CD8, and CD20 staining. Quantification of RNAscope signals was performed using QPath software. Kaplan–Meier survival analysis and multivariate Cox regression were used to assess the prognostic impact of CD70, CD27, and immune cell infiltrates on overall survival (OS). CD70 was expressed in 46

BACKGROUND AND PURPOSE:Hidden cardiotoxicity is defined as drug-induced cardiotoxicity that becomes obvious only in the presence of comorbidities. However, the molecular mechanisms of hidden cardiotoxicity are not always known. Therefore, unbiased multi-omics approaches could assist in revealing regulatory pathways. The most notable representative of hidden cardiotoxic drugs is the cyclooxygenase-2-inhibitor, rofecoxib. We previously reported increased mortality in rats because of proarrhythmic effects of rofecoxib in ischaemic hearts. Here, we aimed to identify molecular mechanisms of hidden cardiotoxicity exemplified by rofecoxib that present prior to comorbidities. EXPERIMENTAL APPROACH:Rats were treated with rofecoxib or its vehicle for 4 weeks. RNA sequencing and proteomic datasets of heart samples were used for differential expression and pathway reconstruction analyses. KEY RESULTS:In this model, mechanisms of hidden cardiotoxicity could not be revealed by transcriptomic analyses. However, mass-spectrometry-based proteomics showed conspicuous changes, revealing 132 proteins that were dysregulated in expression or on phosphorylation sites. Importantly, the phospho-proteomics allowed us to identify two kinases that may mediate cardiotoxicity. Finally, pathway reconstruction maps a complex molecular machinery whose clustered proteins regulate processes involving cytoskeleton binding, mRNA processing, proteolysis, translation, citrate acid cycle and calcium ion signalling. CONCLUSION AND IMPLICATIONS:This is the first demonstration that multi-omics characterisation can reveal underlying regulatory pathways of hidden cardiotoxicity. Importantly, our study shows that transcriptomics gives limited information on the hidden cardiotoxic effects of rofecoxib, which are mainly mediated by changes in posttranslational modifications and protein expression. These changes, among other mechanisms, may disturb the cardiac calcium handling, which could explain the fatal arrhythmias following ischaemia/reperfusion observed with rofecoxib.

Hidden cardiotoxicity is defined as cardiotoxicity of a drug that manifests only in the diseased heart. We have previously shown that the proarrhythmic hidden cardiotoxic properties of a model drug, the selective cyclooxygenase-2 inhibitor rofecoxib, can be revealed in preclinical models of ischemia/reperfusion injury. As metabolic comorbidities, such as hypercholesterolemia (HC), may exacerbate hidden cardiotoxicity, we aimed to investigate the hidden cardiotoxic effects of the model drug, rofecoxib, in the presence of hypercholesterolemia. Rats were fed a high-cholesterol diet for 12 weeks and treated with 5.12 mg/kg rofecoxib. Four weeks of rofecoxib treatment surprisingly improved HC-induced mild cardiac dysfunction by restoring end-diastolic pressure, stroke work, and mechanical efficiency. Then, RNA sequencing revealed that the expression of 28 miRNAs and 300 genes was significantly altered in the HC-fed group. The HC-induced expression changes of miR-27a-5p and miR-30d-5p were reversed by rofecoxib treatment. Cdc42ep4, Cox5, and Cxcl9 genes were also counter-regulated following rofecoxib treatment compared to HC-induced changes. This is the first demonstration that rofecoxib improves HC-induced cardiac dysfunction, with the mechanism involving changes in the gene expression profile, including some key regulators of rofecoxib action.

Background Obesity is a major risk factor for the development of cardiovascular disease. However, recent research shows that moderate obesity reduces the risk of developing cardiovascular disease. We evidenced before that MAO-B inhibitor selegiline reduced visceral adiposity. Aim Therefore, our aim was to investigate cardiac effects of selegiline in moderate obesity in rats treated with a high-fat diet (HFD). Key Findings We demonstrated that HFD improved cardiac contractility parameters, which were reversed by selegiline. Enhanced contractility might be attributed to an increased sarcoplasmic/endoplasmic reticulum Ca2+-ATPase (SERCA2a) expression and phospholamban pentamerization. Selegiline reduced SERCA2a expression in HFD. HFD increased Tumor necrosis factor and Nuclear factor-kappa B expression which were not affected by selegiline. HFD induced proapoptotic processes, which were restored by selegiline. Conclusion In conclusion, moderate obesity improves cardiac function through Ca2+ homeostasis and inflammatory processes and MAO-B inhibition reverses these effects.

Background/Objectives: Small cell lung cancer (SCLC) is one of the malignancies with the worst prognosis, and there have been no major breakthroughs in its treatment for a long time. The majority of patients are diagnosed at the extensive stage, where the only option is chemotherapy, and even the addition of immune checkpoint inhibitors results in only modest benefits. The characterization of the molecular mechanisms behind therapy resistance has relevance in finding novel therapeutic approaches. Previous studies showed the possibility of annexin A1’s (ANXA1) involvement in the immunosuppressive tumor microenvironment in SCLC, and there are studies showing the direct effects of ANXA1 modulation on cancer cell aggressiveness. Methods: We aimed to characterize the roles of ANXA1 expression using publicly available transcriptomic data, the RNA-seq-based predictive algorithms EPIC and ESTIMATE, and immunohistochemistry on patient samples. For the in vitro studies, we silenced ANXA1 expression with short hairpin RNA in three SCLC cell lines, measured the growth rate with the trypan blue exclusion assay, assessed the chemosensitivity to cisplatin and etoposide with the Presto BlueTM viability assay, and performed Western blots to assess changes in the levels of metabolic and mesenchymal markers and transcriptional drivers. Results: ANXA1-high tumors are associated with significantly increased immune infiltrates, stromality, and tumor-associated macrophages (TAMs). The ANXA1 protein is expressed on tumor cells and TAMs at the tissue level. ANXA1 silencing in H841 cells did not affect the growth rate; in SW1271 cells, shANXA1 cells grew significantly slower than shCTRL cells. Meanwhile, in H1048 cells, proliferation was significantly faster. Despite the different growth rates of the tested cell lines, ANXA1 silencing decreased the chemosensitivity to both cisplatin and etoposide in all three cell lines. Gene expression changes in mesenchymal markers, metabolic markers, dominant transcriptional drivers, and immune-relevant molecules were also characterized. Conclusions: This is the first comprehensive characterization of ANXA1 in SCLC to reveal its role in the tumor’s cell biology and the TME, aiming to boost further research in the field.

BACKGROUND:The flipped classroom (FC) approach has demonstrated efficacy in enhancing the learning process, including within higher education contexts such as medical education. Recently, FC has emerged as potential alternative to traditional teaching models across various disciplines, particularly due to its more engaging nature. However, there is limited data available regarding its impact on student performance, particularly in the context of long-term knowledge retention in pharmacology. In this study, our objective was to assess the short- and long-term impact of FC on student performance in Hungarian pharmacology teaching at medical faculty in Semmelweis University (Budapest, Hungary). METHODS AND RESULTS:161 medical students and 10 teachers were involved in this study. We flipped four seminars, then we assessed the academic performance by using multiple choice tests immediately and two weeks after the flipped class. A follow-up assessment was conducted six months after the initial two FC sessions. Our findings indicated that the FC approach enhanced both short- and long-term knowledge retention across most topics. Notably, this long-term improvement was evident even six months after the original seminars on specific subjects. However, despite these topic-specific benefits, the overall performance, including exam grades, did not show significant improvement when compared to the conventional teaching approach. Additionally, we assessed student and teacher perceptions using two questionnaires immediately after practice and at the end of the year. According to the questionnaire responses, students perceived positively the FC approach, emphasizing its interactive and thought-provoking aspects. However, they identified the time-consuming nature of preparation as a significant concern. Teachers also viewed the FC approach favorably, particularly appreciating its interactivity and potential for greater effectiveness. Interestingly, more experienced teachers were less receptive to the FC method, and their perceptions of it were less favorable compared to those of their younger colleagues. CONCLUSIONS:The flipped classroom approach presents a viable strategy for teaching pharmacology, with the potential to enhance student performance and engagement. However, student occupation and faculty resistance may pose significant challenges to the implementation of such alternative teaching methods.

Background and purposeMicroRNA (miRNA) therapy is a promising approach to induce cardioprotection. We have previously identified cardiac microRNA-125b* (microRNA-125b-2-3p; miR-125b*) as a potential cardioprotective miRNA, termed ProtectomiR. We aimed to characterize the pharmacokinetics and pharmacodynamics, and the effect of miR-125b* mimic on infarct size using an in vivo mouse model.Experimental approachTo characterize the pharmacokinetics properties of miR-125b* mimic, a single injection of 10-mu g miR-125b* mimic or its scramble miRNA control, or vehicle i.v. was given to C57BL/6 mice. MiR-125b* expression was measured from plasma, heart, kidney and liver samples. Effect of miR-125b* on area at risk and infarct size was assessed after 45-min coronary occlusion, followed by 24-h reperfusion; 10-mu g miR-125b* mimic or 10-mu g non-targeting miRNA mimic control or vehicle were administered via the right jugular vein at 10th mins of coronary occlusion. To assess molecular mechanism involved in cardioprotection, expression of mRNA targets of miR-125b* were measured from ventricular myocardium at 1, 2, 4, 8 or 24 h post-treatment using quantitative real time polymerase chain reaction.Key resultsMiR-125b* expression was markedly increased in plasma and myocardium 1 h, and in the liver 2h after treatment. Infarct size was significantly reduced after miR-125b* mimic treatment when compared to the vehicle. The expression of Ccna2, Eef2k and Cacnb2 target mRNAs was significantly reduced 8 h after injection of miR-125b* mimic.Conclusion and implicationsThis is the first demonstration of pharmacokinetic and molecular pharmacodynamic properties as well as the cardioprotective effect of miR-125b* mimic in vivo.

Az evidenciaalapú orvoslás alapvető módszerei közé tartozik a gyógyszeres terápia. A 2005. évi XCV. törvény szerin gyógyszernek nevezett „bármely anyag vagy azok keveréke, amelyet emberi betegségek megelőzésére vagy kezelésére alkalmazható termékként jelenítenek meg, vagy azok az anyagok vagy keverékei, amelyek farmakológiai, immunológiai vagy metabolikus hatások kiváltása révén az ember valamely élettani funkciójának helyreállítása, javítása vagy módosítása, illetve az orvosi diagnózis felállítása érdekében az emberi szervezetben vagy emberi szervezeten alkalmazhatók” (1). Gyakorlatilag nincs olyan szakterület, ahol a gyógyszerek alkalmazása kikerülhető. A betegek nem egyformán reagálnak a gyógyszeres kezelésre, sőt, a nemkívánatos hatások kialakulási valószínűsége sem ugyanaz mindenkinél. Az eltéréseknek számos oka lehet, ezeket foglalja össze az 1. táblázat (2).

Background and Purpose: Cardioprotective miRNAs (protectomiRs) are promising therapeutic tools. Here, we aimed to identify protectomiRs in a translational porcine model of acute myocardial infarction (AMI) and to validate their cardiocytoprotective effect. Experimental Approach: ProtectomiR candidates were selected after systematic analysis of miRNA expression changes in cardiac tissue samples from a closed-chest AMI model in pigs subjected to sham operation, AMI and ischaemic preconditioning, postconditioning or remote preconditioning, respectively. Cross-species orthologue protectomiR candidates were validated in simulated ischaemia-reperfusion injury (sI/R) model of isolated rat ocardiomyocytes and in human AC16 cells as well. For miR-450a, we performed target prediction and analysed the potential mechanisms of action by GO enrichment and KEGG pathway analysis. Key Results: Out of the 220 detected miRNAs, four were up-regulated and 10 were down-regulated due to all three conditionings versus AMI. MiR-450a and miR-451 mimics at 25 nM were protective in rat cardiomyocytes, and miR-450a showed protection in human cardiomyocytes as well. MiR-450a has 3987 predicted mRNA targets in pigs, 4279 in rats and 8328 in humans. Of these, 607 genes are expressed in all three species. A total of 421 common enriched GO terms were identified in all three species, whereas KEGG pathway analysis revealed 13 common pathways. Conclusion and Implications: This is the first demonstration that miR-450a is associated with cardioprotection by ischaemic conditioning in a clinically relevant porcine model and shows cardiocytoprotective effect in human cardiomyocytes, making it a promising drug candidate. The mechanism of action of miR-450a involves multiple cardioprotective pathways.

Introduction: Ischemic conditionings (ICon) were intensively investigated and several protective signaling pathways were identified. Previously, we have shown the role of matrix metalloproteinases (MMP) in myocardial ischemia/reperfusion injury (MIRI) and the cardioprotective role of biglycan (BGN), a small leucine-rich proteoglycan in vitro. Here, we hypothesized that cardiac MMP and BGN signaling are involved in the protective effects of ICon. Methods: A reverse target-microRNA prediction was performed by using the miRNAtarget™ 2.0 software to identify human microRNAs with a possible regulatory effect on MMP and BGN, such as on related genes. To validate the identified 1289 miRNAs in the predicted network, we compared them to two cardioprotective miRNA omics datasets derived from pig and rat models of MIRI in the presence of ICons. Results: Among the experimentally measured miRNAs, we found 100% sequence identity to human predicted regulatory miRNAs in the case of 37 porcine and 24 rat miRNAs. Upon further analysis, 42 miRNAs were identified as MIRI-associated miRNAs, from which 24 miRNAs were counter-regulated due to ICons. Conclusions: Our findings highlight 24 miRNAs that potentially regulate cardioprotective therapeutic targets associated with MMPs and BGN in a highly translatable porcine model of acute myocardial infarction.

The pharmaceutical medicine course at the Semmelweis University of Budapest, Hungary, was initiated as part of the Innovative Medicines Initiative (IMI is the main program, IMI-PharmaTrain is one of the IMI projects) Pharmaceutical Medicine Training Programs (16 IMI Call 2008/1/16). The aim was to extend training in the development of pharmaceutical medicine to those EU member states where no such education was present. The final program envisaged the development of a cooperative education supported by universities located in Central and Eastern Europe. It was considered to be the economically and scientifically most viable approach to combine the expertise from these countries to form a united teaching staff and provide education jointly for young professionals of the region. Semmelweis University was selected to manage this coordinated program. In this report, we describe the organization and functioning of this international university-based pharmaceutical medicine education project called the Cooperative European Medicines Development Course (CEMDC) and evaluate its successes and shortcomings. During the pandemic, the educational course was interrupted. The follow-on program is reorganized as a postgraduate MSc course named “Semmelweis Pharma MBA” and will be started in 2025. It will continue the established PharmaTrain educational tradition. However, it will deal in more detail with the transition from basic pharmacological to industrial research, as well as biopharmaceutical formulation and manufacturing and marketing aspects of medicines development.

Lipid-lowering drugs have been shown to have cardioprotective effects but may have hidden cardiotoxic properties. Therefore, here we aimed to investigate if chronic treatment with the novel lipid-lowering drug bempedoic acid (BA) exerts hidden cardiotoxic and/or cardioprotective effects in a rat model of acute myocardial infarction (AMI). Wistar rats were orally treated with BA or its vehicle for 28 days, anesthetized and randomized to three different groups (vehicle + ischemia/reperfusion (I/R), BA + I/R, and positive control vehicle + ischemic preconditioning (IPC)) and subjected to cardiac 30 min ischemia and 120 min reperfusion. IPC was performed by 3 × 5 min I/R cycles before ischemia. Myocardial function, area at risk, infarct size and arrhythmias were analyzed. Chronic BA pretreatment did not influence cardiac function or infarct size as compared to the vehicle group, while the positive control IPC significantly reduced the infarct size. The incidence of reperfusion-induced arrhythmias was significantly reduced by BA and IPC. This is the first demonstration that BA treatment does not show cardioprotective effect although moderately reduces the incidence of reperfusion-induced arrhythmias. Furthermore, BA does not show hidden cardiotoxic effect in rats with AMI, showing its safety in the ischemic/reperfused heart.

A common way to investigate epilepsy and the effect of antiepileptic pharmaceuticals is to analyze the movement patterns of zebrafish larvae treated with different convulsants like pentylenetetrazol (PTZ), pilocarpine, etc. Many articles have been written on this topic, but the research methods and exact settings are not sufficiently defined in most. Here we designed and executed a series of experiments to optimize and standardize the zebrafish epilepsy model. We found that during the light and the dark trials, the zebrafish larvae moved significantly more in the light, independent of the treatment, both in PTZ and pilocarpine-treated and the control groups. As expected, zebrafish larvae treated with convulsants moved significantly more than the ones in the control group, although this difference was higher between the individuals treated with PTZ than pilocarpine. When examining the optimal observation time, we divided the half-hour period into 5-minute time intervals, and between these, the first 5 minutes were found to be the most different from the others. There were fewer significant differences in the total movement of larvae between the other time intervals. We also performed a linear regression analysis with the cumulative values of the distance moved during the time intervals that fit the straight line. In conclusion, we recommend 30 minutes of drug pretreatment followed by a 10-minute test in light conditions with a 5-minute accommodation time. Our result paves the way toward improved experimental designs using zebrafish to develop novel pharmaceutical approaches to treat epilepsy.