University of Zagreb

Vaccination against lumpy skin disease was carried out in Croatia during 2016 and 2017 to prevent this emerging viral disease from causing significant damage to the cattle population. Two unauthorised live vaccines were used for immunoprophylaxis, and adverse reactions in cattle were expected. Adverse events were monitored and 14 selected reported cases of deaths were analysed using the available necropsy results. The cases analysed were classified as probable, possible, inconclusive, or unlikely to be caused by vaccination, primarily based on results of post-mortem examination, taking into consideration factors that contributed to or resulted in death or euthanasia. Most cases were classified as possibly caused by vaccination due to subcutaneous oedema of varying consistency (at the injection site) and intramuscular haemorrhage, in some cases leading to purulent and necrotic processes. These processes eventually led to fatal purulent bronchopneumonia, pulmonary emphysema, septic conditions, or euthanasia. In the remaining cases, necropsy results were unable to be linked to vaccination or the findings were inconclusive due to the presence of other factors, such as Klebsiella infection. It can be assumed that neither vaccine had a direct connection with animal deaths, but they may have contributed to the fatal outcomes due to the complications of severe local reactions and general impact on the immune system.

Mitochondrial calcium ([Ca2+]mito) homeostasis is a key regulator of cellular physiology, controlling signal transduction, energy metabolism, and cell survival. To examine how these processes change with age in vivo, we used the pharyngeal muscle of Caenorhabditis elegans (C. elegans) as a tractable model for studying [Ca2+]mito dynamics. We introduced electric field stimulation as a robust trigger for [Ca2+]mito uptake that overcomes limitations of compound stimulation, which relies on pharyngeal pumping and endoplasmic reticulum store filling and is typically confined to single-stimulus protocols. In contrast, electric field stimulation enables physiologically relevant repeated excitation that challenges [Ca2+]mito uptake and recovery. Stimulation with 10V for 10s reliably evoked reproducible [Ca2+]mito transients in the pharyngeal muscle of C. elegans and produced higher responder rates than compound stimulation. Basal [Ca2+]mito increased with age, and the first field-evoked transient was significantly larger in aged animals, an effect not detected with pharmacological triggers. Repeated pulses unmasked cumulative [Ca2+]mito loading and incomplete recovery in aged pharynx, indicative of [Ca2+]mito overload, which was attenuated by inhibition of the mitochondrial Ca2+ uniporter (MCU) with mitoxantrone. Regional analyses identified the corpus as a hotspot for electric field-evoked [Ca2+]mito uptake and overload, while MCU inhibition reduced repeated responses in both corpus and posterior bulb. Electric field stimulation enables precise, repeated in vivo probing of [Ca2+]mito uptake and recovery, revealing overload in the C. elegans pharynx. This approach identified age-enhanced, MCU-dependent, and region-specific [Ca2+]mito loading, providing a pathophysiologically relevant readout of impaired Ca2+ handling that may contribute to age-related muscle dysfunction.

Oilseed cakes are by-products of oil extraction and they have considerable potential in novel food technologies due to their low cost and high nutrient content. In vitro production of skeletal muscle cell biomass is an emerging approach in cultured meat research and offers a promising alternative protein source. One of the major barriers to large-scale production is the reliance on animal serum, an expensive, animal-derived component. To address this limitation, this study explored the potential of flaxseed protein isolate (FPI), extracted from flaxseed oil cake, as a supplement in cell culture media with a fourfold reduction in serum for the cultivation of C2C12 myoblasts. Flaxseed protein isolate was evaluated for its impact on cell proliferation and differentiation in low-serum media. As the media component, FPI did not enhance C2C12 cell proliferation, but it facilitated myogenic differentiation significantly. Specifically, supplementing low-serum media with 0.25 mg mL-1 FPI supported cell survival throughout the 10 day differentiation period, resulting in typical myogenic marker presence. It also promoted the formation of myotubes with diameters and fusion index comparable with those observed in control cell cultures with standard serum level. Flaxseed protein isolate also considerably increased the production of sulfated glycosaminoglycans (sGAGs), an extracellular matrix component, exceeding even the levels observed in the control group. The results of this study suggest that flaxseed protein may serve as a valuable supplement in low-serum media to support myoblast differentiation. This could help reduce the cost of cultured meat production while supporting circular economy principles and the reuse of agricultural byproducts. © 2026 Society of Chemical Industry.

Background: Balkan nephropathy (BEN) is an environmental form of aristolochic acid nephropathy (AAN) strongly associated with upper urinary tract urothelial carcinoma (UTUC). Clinical diagnosis remains challenging, and misclassification is frequent. The study reassessed BEN diagnoses in kidney transplant recipients from rural farming areas who did not undergo prophylactic bilateral nephroureterectomy and evaluated post-transplant outcomes. Methods: In this retrospective single-centre study, we analysed 12 kidney transplant recipients from rural Balkan farming regions. BEN diagnoses were reevaluated according to international consensus criteria. Key endpoints included patient and graft survival, post-transplant clinical course, and UTUC characteristics. Results: Upon reassessment, three of the six patients initially diagnosed with BEN were reclassified as having BEN. Among the remaining six patients, four were reclassified as having sporadic BEN. Overall, only 33% of patients had a diagnosis concordant with their admission records. During a median follow-up of 6.8 years (IQR 2.1-15.8), UTUC developed in seven out of 12 patients. The UTUC cases were predominantly high-grade and multifocal, and they were identified as the leading cause of death. Five-year patient and graft survival rates were 71% and 100%, respectively. Conclusions: BEN is frequently misdiagnosed or misclassified in kidney transplant candidates from rural farming areas. Despite excellent graft survival, the high incidence of post-transplant urothelial carcinoma underscores the necessity of accurate diagnosis and the consideration of prophylactic bilateral nephroureterectomy. Lifelong intensive surveillance is essential not only for patients from established BEN regions but also for individuals from other rural farming areas at risk for sporadic BEN.

Equal Channel Angular Pressing (ECAP) is a widely used severe plastic deformation technique for producing ultrafine-grained metals. Numerical modelling plays an essential role in analyzing the process mechanics. The purpose of this study is to reduce the computational cost of explicit ECAP simulations by applying mass scaling while ensuring that the accuracy of the results is not compromised. The methodology includes explicit finite-element modelling with controlled mass scaling and verification by monitoring internal, kinetic, and total energies throughout deformation. The ratio of kinetic to internal energy is used as the primary indicator of acceptable quasi-static behavior. The results show that the applied mass-scaling level significantly reduces computational time without altering the predicted stress distribution, plastic strain fields, or forming forces, provided that the kinetic energy remains within recommended limits. The application of mass scaling reduced simulation time by more than two orders of magnitude while preserving quasi-static conditions, as indicated by a consistently low kinetic-to-internal energy ratio. The study concludes that energy-based verification offers a reliable framework for applying mass scaling in ECAP simulations while preserving the physical validity of the results.

Landau level spectroscopy plays an important role in modern condensed-matter physics. In this technique, electrons in a solid are subjected to quantising magnetic fields and probed experimentally, often through optical methods. Direct and detailed insights into the electronic properties of crystalline materials are obtained, particularly the properties related to their band structure. Landau level spectroscopy enables the precise extraction of key parameters such as effective mass, carrier density, mobility, and band gap, and serves as a powerful tool for studying interactions between electrons and other quasiparticles in solids. Over its more than seventy-year history, Landau level spectroscopy has been applied mainly to semiconductors and semimetals. Today, its scope also includes graphene-based systems, surface and bulk states in topological materials, and other emergent systems with a narrow or vanishing band gap. In this work, we review the fundamentals of Landau level spectroscopy and illustrate them with selected examples from the literature.

Electric vehicles (EVs) experience substantially reduced driving range in cold weather, primarily due to cabin heating energy demands. This paper proposes a control allocation strategy for positive temperature coefficient (PTC) heater-based electric minibus cabin thermal management, aimed at minimizing energy consumption. The strategy is of a hierarchical structure, where a supervisory PI cabin temperature controller commands the heating power demand, which is then achieved through optimal allocation and low-level control of the cabin inlet air temperature, coolant pump flow, and radiator blower air flow control inputs. Based on the assumption of fast heating system dynamics relative to cabin thermal dynamics, quasi-steady-state optimization of control input allocation is carried out by employing a grid-search algorithm over a dataset resulting from high-fidelity simulations. For the system heat-up transient conditions, where the steady-state allocation proves to be suboptimal, dynamic programming is applied on a validated reduced-order model to optimize the control trajectories. Insights gained through control trajectory optimization are then used to develop a rule-based modification of the control allocation strategy for the heat-up scenario. Simulation verification of the overall control system demonstrates energy consumption reduction in the range from 4 to 12% when compared to the industrial baseline system across both steady-state and transient operating conditions.

Abstract Once nearly eradicated from Europe, grey wolves ( Canis lupus ) have recently undergone a remarkable demographic recovery, but their long-term survival remains precarious. By analyzing 1,001 genomes, we uncover a mosaic of distinct evolutionary lineages, not a single recovering population. Northern populations exhibit Asian wolf ancestry, while southern populations preserve ancient Holocene lineages, with dog introgression varying by region. Isolated wolves from the Scandinavian, Italian, and Iberian peninsulas harbored particularly high levels of inbreeding and fixed deleterious mutations. Signatures of genome erosion were widespread across Europe, with many populations falling well short of the minimum size recommended for long-term survival. Our study reveals the complex tapestry of wolf ancestry and variation across Europe, which calls for nuanced, regional conservation plans founded in genetic monitoring.