Institute of Clinical and Experimental Medicine

Donor kidney tissue based transcriptomics may represent new dimension for prediction of kidney transplant outcomes. In this prospective, single-center study, 276 kidneys from 174 deceased brain-death donors were assessed by microarrays to identify phenotypes of procurement biopsies. Molecular classifiers (extreme gradient boosting, logistic and Poisson regression) with 10-fold cross-validation were employed to categorize donors based on clinical variables (age, BMI, hypertension, ECD kidney) and histological scores (vascular fibrous intimal thickening, interstitial fibrosis, tubular atrophy, arteriolar hyaline thickening). Archetypal analysis and linear mixed model were applied to determine molecular phenotypes and their association with posttransplant 1-year eGFR in 234 donor kidneys. Three molecular archetypes were identified. The "ideal" archetype (median donor age 42 years, low KDRI, minimal chronic histological changes) was associated with the highest 1-year eGFR, while the "marginal" archetype (68 years, extensive chronic changes, high KDRI) with the lowest one. The "intermediate" archetype yielded better 1-year eGFR despite donor profiles similar to the marginal group. While KDRI predicted 1-year eGFR, adding molecular archetypes improved model performance (AIC 80.0 vs. 83.7;p<0.05). External validation in an independent dataset (n=174, GSE147451) confirmed predictive value of the model. Molecular profiling of procurement biopsies may help to identify donor kidneys with higher posttransplant eGFR.

Wild-type transthyretin cardiac amyloidosis (ATTRwt-CA) is an increasingly recognized disease in elderly individuals. We aimed to perform a cross-sectional survey investigating the current diagnostic journey of ATTRwt patients in the Czech Republic from initial symptoms to final diagnosis. Between November 2022 and May 2023, most data were prospectively obtained using a dedicated questionnaire from 118 ATTRwt-CA patients during the regular outpatient consultation. The rest of the data were taken by attending physicians from medical records. The mean age of the patients at the time of diagnosis was 77 ± 6 years, 85.6% were males. The time interval from the onset of clinical scenario leading to suspicion of CA to suspected diagnosis was 14±24 months, while the subsequent period to definite ATTRwt-CA diagnosis was 2.5±2.3 months. Heart failure primarily led to the suspicion of CA in 61.9%, followed by suspicious echocardiographic findings (12.7%) and atrial fibrillation or flutter (6.8%). The history of carpal tunnel syndrome was the most common extracardiac manifestation found in 53.9% subjects, with median duration of 99 months. The diagnosis of CA was most often first suspected by outpatient cardiologists (33.1%) and cardiologists working at a cardiocenter (25.4%). At the time of diagnosis, NYHA class I or II was present in 10% and 56% of patients, respectively. The definite ATTRwt-CA diagnosis was made noninvasively in 63% cases. A relatively long-time delay still exists between the onset of clinical manifestations and suspicion of CA. Nevertheless, once CA is suspected, the definite ATTRwt-CA diagnosis is made early. Although heart failure still represents the most common clinical scenario, several other clinical profiles lead quite frequently to suspicion of the disease. The non-invasive diagnostics of ATTRwt-CA is done in majority of our patients, who are diagnosed mostly in early stages of the disease. The awareness of ATTRwt manifestations must be increased in order to shorten the whole diagnostic process of the disease.

Atherosclerosis is a chronic, progressive disease of the arterial wall and the principal pathological substrate underlying most cardiovascular diseases, including ischemic heart disease, stroke, and peripheral arterial disease. Despite advances in prevention, imaging, and therapy, atherosclerosis remains the leading cause of cardiovascular morbidity and mortality worldwide. From a pathological perspective, the disease represents a dynamic and heterogeneous process characterized by endothelial dysfunction, lipid retention and modification, chronic inflammation, immune activation, smooth muscle cell phenotypic modulation, extracellular matrix remodeling, and thrombogenic surface alterations. This review provides a comprehensive overview of atherosclerosis from a pathologist's perspective, integrating classical morphological concepts with contemporary insights into immunopathology, plaque classification, and mechanisms of plaque instability. We summarize the structure and function of the arterial wall, the stepwise pathogenesis of lesion initiation and progression, and the histopathological classification systems established by the American Heart Association and subsequently refined through Virmani's framework. Particular emphasis is placed on plaque instability, highlighting the qualitative features-such as fibrous cap thinning, necrotic core expansion, macrophage-driven inflammation, plaque erosion, and calcification patterns-that determine clinical outcomes rather than luminal stenosis alone. Furthermore, the review discusses the expanding role of immunohistochemical markers in defining plaque biology, including lineage markers and functional indicators of inflammation, matrix integrity, osteogenic signaling, and local anticoagulant balance. These pathological insights are integrated with contemporary risk assessment tools, imaging modalities, preventive strategies, and therapeutic interventions, including emerging lipid-lowering and RNA-based therapies. In conclusion, pathology remains central to understanding atherosclerosis as a biologically active disease and to refining concepts of plaque instability. Integrating histopathology with molecular profiling, imaging, and clinical data is essential for advancing precision prevention and targeted treatment strategies in atherosclerotic cardiovascular disease.

The molecular mechanisms guiding cochlear development remain incompletely understood. Here, we used conditional deletion mouse models (both sexes) to define the spatiotemporal roles of the transcription factors NEUROD1 and ISL1 during cochlear neurosensory development. Delayed deletion of Neurod1 using Atoh1Cre had no detectable effect on cochlear structure or hearing function. In contrast, Atoh1Cre-mediated Isl1 deletion disrupted sensory epithelium organization, producing ectopic inner hair cells and supernumerary outer hair cell rows, and altered spiral ganglion development. Although Atoh1Cre targeted only a subset of developing spiral ganglion neurons, Isl1 loss resulted in aberrant spiral ganglion morphology, reduced neuronal density, impaired innervation, and defective neuronal migration. These abnormalities arose through a combination of cell-autonomous effects in Isl1-deficient neurons and non-cell-autonomous effects on untargeted, neighboring neurons. The resulting neurosensory abnormalities compromised hearing function, evidenced by elevated auditory brainstem response thresholds, abnormal low-frequency acoustic stimulus processing, and reduced distortion product otoacoustic emissions. Together, these findings further substantiate the essential role of ISL1 in auditory neuron development and establish ISL1 as a key regulator of cochlear neurosensory development.

Fluorine-19 magnetic resonance imaging (19F MRI) is a powerful imaging modality that provides background-free imaging. Its sensitivity can be modulated through environmentally responsive probe designs exploiting paramagnetic relaxation enhancement (PRE) and related mechanisms. Here, we present a novel pH-sensitive 19F MRI nanosystem in which signal activation is governed by nanoparticle (NP) ultrastructure. Poly(lactic-co-glycolic acid) (PLGA) NPs were synthesized co-encapsulating perfluoro-15-crown-5-ether (PFCE) and lipophilic gadolinium (Gd) chelates, yielding either single-core or multi-core structures. While single-core NPs, the conventional structure for polymer-encapsulated perfluorocarbons, remained pH-insensitive, multi-core NPs exhibited a pronounced and reversible pH-dependent 19F signal modulation. The 19F T1 decreased slightly (from ∼700 to ∼600 ms) in both NP types due to the presence of Gd, whereas T2 shortened markedly at neutral pH (∼20 ms, “OFF” state) and increased substantially under acidic conditions (“ON” state); this variation was absent in single-core NPs. The pH-dependent 19F T2 behaviour was consistent across both high-field (14 T) and low-field (3 T) MRI. Such behaviour of multi-core NPs was validated in vitro and in vivo, as the 19F signal reappeared only after lysosomal internalization in RAW 264.7 cells and was selectively activated within acidic tumour regions, but not at the neutral subcutaneous injection site. By exploiting nanoparticle ultrastructure to achieve reversible signal switching, this work introduces a general strategy for activatable 19F MRI probes for imaging applications in tumours and other pH-sensitive disease environments. It demonstrates, for the first time, PRE-driven signal enhancement and suppression governed by structural rearrangement rather than chemical modification in response to pH variation.

In pharmacology and clinical pharmacology (P&CP) teaching, using generic names of medicines contributes to global comprehensibility. However, brand names are often used in routine clinical practice, which may pose a challenge in the educational setting. This study aimed to explore P&CP teachers’ use of and attitudes towards generic and brand names in undergraduate education, as well as national prescription regulations. A survey was conducted with P&CP teachers from 38 European countries. An electronic questionnaire was used to collect data on the participants’ profiles, the nomenclature used and preferred in undergraduate education, and prescribing practices. The latter were then verified with national regulatory authorities. Sixty-one teachers from 23 countries participated in the study, the majority being physicians (n = 47). In total, 13 (21%) teachers use only generic names in their teaching, while 46 (75%) use both generic and brand names. Thirty-four (56%) teachers preferred listing brand names in case-based teaching, as this reflects real life. Conversely, 37 (61%) teachers stated that students should not be exposed to brand names before graduation. Generic prescribing is either allowed or mandatory in all participating countries except two, where brand-name prescribing is mandatory. This study shows that generic names must remain fundamental in P&CP education, but brand names also play a role, especially in case-based education, reflecting real-life practice. Potential implications of generic and brand prescribing for internationalisation in education could be worth further attention.

Abstract Background Comparative data on risankizumab (RISA) and upadacitinib (UPA) in Crohn’s disease (CD) are limited. In this preliminary analysis, we assessed treatment persistence with these agents and explored whether the choice should be modified according to disease location. Methods Data from the Czech National Prospective Registry of Biological and Innovative Therapies for Inflammatory Bowel Disease (CREdIT) were analysed to evaluate treatment persistence in adult and paediatric patients with CD who began treatment with RISA or UPA, with a minimum follow-up period of three months. Patients with missing outcomes or dual advanced therapy were excluded. Inverse-probability-of-treatment weighting (IPTW) based on a propensity score including demographic, disease activity and treatment history variables was applied to reduce confounding factors. Treatment persistence was analysed using Kaplan–Meier curves and Cox proportional hazards models, clustered by patient. We evaluated whether disease location (ileal vs non-ileal) altered treatment persistence by using group-specific survival analyses. Results A total of 474 treatment observations were identified in patients receiving RISA or UPA. Following the application of exclusion criteria, the analysis included 370 observations from 323 patients, of which 157 (42%) were on RISA and 213 (58%) were on UPA. Of these, 11 (7%) were paediatric patients in the RISA group and 22 (10%) in the UPA group. The median number of actual lines of advanced therapy was four in both groups, with 38% of RISA observations and 39% of UPA observations representing ≥ 5th treatment line. Groups differed only in age at treatment start (median 38 vs 37 years). Ileal disease only was present in 23 (15%) RISA patients and 28 (13%) UPA patients. Unadjusted data (Figure 1) demonstrated higher persistence of RISA vs. UPA treatment. After balancing the cohort using IPTW, all standardised mean differences between groups were below 0.1. Compared with RISA, UPA was associated with a higher risk of treatment discontinuation (hazard ratio: 2.83; 95% confidence interval: 1.50–6.34). Interaction analysis by disease location was not feasible because no RISA failures occurred in the ileal subgroup. Therefore, only descriptive stratified curves are shown in Figure 2. Conclusion In this preliminary analysis of refractory CD, treatment persistence was greater with RISA than with UPA. These real-world data suggest that RISA may be a more durable treatment option, particularly for patients with ileal CD. However, confirmation in larger patient groups is required. Conflict of interest: Dr. Hradsky, Ondrej: Lectures/congress fees/consultancy from MSD, AbbVie, Takeda, Sandoz, Nutricia, Ferring, Pfizer, and Lilly. Duricova, Dana: Lectures/congress fees/consultancy (outside the scope of the submitted work) from Johnson &amp; Johnson, Takeda, AbbVie, Pfizer, Ferring, Eli Lilly, and Celltrion. Lukás, Milan: Research Support / Clinical trials: Biogen, Takeda, Janssen Consultant / Speaker : Abbvie, Ferring, Takeda, Janssen Speakers Bureau / Advisory Board Member: Pfizer, Roche, Egis, Celltrion, Takeda, Janssen, Eli Lilly Employee: ISCARE a.s./1stMedical Faculty, Charles University, Prague Nedbalova, Lenka: No conflict of interest Bouchner, Luděk: No conflict of interest Pipek, Barbora: Lectures/congress fees/consultancy (outside the scope of the submitted work) from Johnson &amp; Johnson, Takeda, AbbVie, Pfizer, and Eli Lilly. Svoboda, Pavel: Lectures/congress fees/consultancy (outside the scope of the submitted work) from AbbVie, Takeda, Janssen-Cilag, Celltrion, Tillotts, Alfasigma, Sandoz, Pfizer, and Eli Lilly. Siroky, Milan: No conflict of interest Bronsky, Jiri: Lectures/congress fees/consultancy (outside the scope of the submitted work) from AbbVie, Sandoz, Danone-Nutricia, Nestlé, Sanofi, Pfizer, and Vitabalans. Bortlik, Martin: Lectures/congress fees/consultancy (outside the scope of the submitted work) from AbbVie, Takeda, Janssen-Cilag, Celltrion, Roche, AstraZeneca, Biogen, Tillotts, Ferring, Alfasigma, PRO.MED.CS, Sandoz, Bristol-Myers Squibb, Pfizer, and Swedish Orphan Biovitrum.

Abstract Interferons (IFNs) orchestrate diverse immune responses, but distinguishing individual IFN contributions in human transcriptomic data is challenging due to overlapping interferon-stimulated gene (ISG) signatures and limited cell-type-specific datasets. To address this, we generated a single-cell transcriptomic atlas of IFN responses by stimulating primary human T, B, NK, and CD14 monocytes with IFN-I, IFN-II, and IFN-III. This revealed core and cell-type-specific ISG programs across 13 subsets, highlighting distinct functions of IFNs. We developed an algorithm to separate IFN-I and IFN-II activity in transcriptomic data. Applied to multiple myeloma samples, it showed elevated IFN-I and IFN-II responses, with induction therapy reducing only IFN-I. Extending to multiple disease datasets provided a cross-disease overview of IFN-I and IFN-II activities and revealed increased IFN-II activities in T cells during lupus flares. This resource and the accompanying analytical framework enable dissection of IFN-driven transcriptional programs in a cell-type specific manner in human disease.

Macrophages are critical components of the innate immune system that adapt their function based on local microenvironmental cues. In adipose tissue, macrophage polarization is intimately linked to metabolic regulation and inflammation. In this pilot study, we performed lipidomic profiling to characterize the phospholipid composition of pro-inflammatory (PI-ATM) and non-inflammatory (NI-ATM) macrophages isolated from visceral adipose tissue of living kidney donors. Tissue samples were processed, and macrophages were isolated and sorted using CD14 and CD16 markers. Lipids were extracted by methyl tert-butyl ether, and differences in the lipidome were analyzed using the optimized UHPLC-MS method. PI-ATM exhibited a higher abundance of lipids across various lipid classes than NI-ATM. Phosphatidylethanolamines and ether lipids derived from phosphatidylethanolamines and phosphatidylcholines were most frequently found among the significantly altered lipids. The majority of significantly altered lipid species contained an octadecenoic aliphatic chain. These findings suggest that alterations in membrane lipid composition - particularly ether lipids and phosphatidylethanolamines - may play a pivotal role in macrophage polarization and adipose tissue inflammation. Our study provides novel insights into the lipidomic regulation of macrophage function in human adipose tissue and identifies potential targets for therapeutic intervention in metabolic disorders.