Joanna Zeyland

Salinomycin, an ionophore coccidiostat widely used in livestock production, has attracted growing interest as a potential anticancer agent, although its clinical application remains limited by pronounced organ-specific toxicity. Cannabidiol (CBD), a non-psychoactive cannabinoid with documented cytoprotective properties, is increasingly consumed, making concurrent exposure plausible, yet its interaction remains unexplored. This study evaluated the cytotoxicity of salinomycin (0.39–50 μg/mL), alone and combined with two non-cytotoxic CBD concentrations (1.56 and 3.125 μg/mL), in three human cell lines—embryonic kidney-derived HEK-293, hepatoblastoma-derived HepG2, and neuroblastoma-derived SH-SY5Y—using assays of mitochondrial and lysosomal activity, membrane integrity, total protein content, and DNA synthesis, complemented by ROS measurement, apoptosis/necrosis evaluation, morphology, and an exploratory combination index analysis. CBD modulated salinomycin cytotoxicity in a strictly cell-line-dependent manner. In HEK-293 cells, it attenuated cytotoxicity, as reflected by uniformly higher IC₅₀ values, reduced ROS production, and lower median fluorescence intensities of Hoechst 33342 and propidium iodide, whereas in the tumor-derived HepG2 and SH-SY5Y lines, it enhanced cytotoxicity irrespective of the CBD concentration applied, accompanied by increased ROS generation and more pronounced cell death. As all three lines are immortalized or tumor-derived, these observations describe cell-line-specific responses and cannot be extrapolated to normal renal, hepatic, or neuronal tissue.

The use of ionophore coccidiostats—salinomycin (SAL), lasalocid (LAS), monensin (MON), and narasin (NAR)—in animal production raises toxicity concerns in non-target species through feed contamination and because of their narrow safety margins. Cannabidiol (CBD) has antioxidant and cytoprotective properties that may modify the cellular response to ionophore-induced toxicity. This study examined the effect of CBD on the cytotoxicity of SAL, LAS, MON, and NAR in feline (CRFK) and canine (MDCK) kidney cells. Cells were exposed for 72 h to each coccidiostat, alone or in combination with CBD, and assessed using mitochondrial activity (MTT), lysosomal activity (NRU), total protein content (TPC), and membrane integrity (LDH) assays; the interaction was evaluated by isobolographic analysis. Molecular docking of CBD and the ionophores to human VDAC1 (PDB 6TIQ) was performed in AutoDock Vina 1.2.7, both individually and in simultaneous multi-ligand combinations, with β-NADH, VBIT-4, and SW016789 as reference ligands. All four ionophores induced concentration-dependent cytotoxicity, with distinct, compound- and species-specific profiles between CRFK and MDCK cells. CBD co-exposure produced divergent interactions depending on the drug and cell model, ranging from marked cytoprotection to pronounced synergistic potentiation of toxicity, most notably for SAL and LAS in canine cells. The ionophores occupied the N-terminal helix, and cannabidiol occupied a partially overlapping C-terminal region, from which β-NADH displaced it, suggesting a plausible structural basis for the observed interactions. These findings indicate that ionophore nephrotoxic potential and its modulation by CBD are both concentration- and species-dependent, warranting further investigation using additional in vitro and in vivo models.

Introduction: Veterinary drug residues in the environment threaten public health and animal welfare. Ionophore coccidiostats (lasalocid (LAS), monensin (MON) and narasin (NAR)) are commonly used in animal production. Overdosing, errors in feed mixing or interactions with other drugs can cause nephrotoxicity in animals, which has also been observed in poisoned humans. The toxic effects of veterinary drugs may be mitigated by cannabidiol (CBD), known to be antioxidant and cytoprotective and expected to have a protective effect on kidney cells. The aim of this study was to evaluate the effect of CBD on cytotoxicity induced by ionophore coccidiostats in human embryonic kidney cells (HEK-293). Material and Methods: Cells were exposed to LAS, MON and NAR, either alone or in combination with CBD. Cytotoxicity was assessed using mitochondrial activity, lysosomal activity, total protein content and membrane integrity assays. The action of combination of CBD and the coccidiostats was determined using interaction analysis. Results: The ionophores induced cytotoxicity dose-dependently. Lasalocid was the most toxic, significantly impairing mitochondrial function and membrane integrity at 1 µg/mL. Co-treatment with CBD significantly attenuated the drugs’ toxic effects. Predominantly, antagonistic effects were revealed, indicating a protective effect of CBD. However, an additive effect was observed at high concentrations of the LAS-CBD combination. Conclusion: Cannabidiol exerts a protective effect against coccidiostat nephrotoxicity in human renal cells. This finding suggests its modulation of cellular responses to ionophore-induced cytotoxicity, offering a basis for further in vivo studies on its potential.

Autoimmune myocarditis frequently progresses to inflammatory cardiomyopathy through dysregulated immune–stromal interactions. This study employs single-nuclei RNA-sequencing (snRNA-seq) to profile 46,233 cardiac nuclei from the experimental autoimmune myocarditis (EAM) mouse model at four timepoints: day 0 (healthy), day 14 (inflammation), day 21 (acute inflammation), and day 40 (late cardiac remodelling). Single-nuclei RNA profiling identified 18 transcriptionally distinct cell populations. Global cell–cell communication analysis revealed a dramatic peak of intercellular signalling at day 14 (5907 interactions), with fibroblast subpopulations and macrophages as dominant hubs, followed by partial resolution at day 21 (2264 interactions) and renewed remodelling at day 40 (4862 interactions). Subclustering of the macrophage compartment identified five subpopulations: Mac-TLF, Mac-MHCII, Mac-rMHCII, Mac-ResL, and Classical Monocytes. Tissue-resident macrophages (Mac-TLF, CCR2-) dominated at healthy state (~55%) but were rapidly depleted at day 14, coinciding with a dramatic influx of recruited CCR2⁺ macrophages (Mac-rMHCII), which expanded to over 70% of the compartment and maintained dominance through day 40. At inflammation (day 14), the expanded Mac-rMHCII subpopulation displayed a strongly pro-inflammatory signature (Il1b, Stat2, Parp14, Apoe), and the overall macrophage compartment was enriched for cytokine response, Fc-gamma receptor, and Notch signalling pathways, while downregulating homeostatic and mitochondrial metabolic programmes, potentially contributing to impaired efferocytosis and cardiomyocyte dysfunction. Macrophage-centred communication networks expanded markedly at day 14 (1047 interactions), with resting fibroblasts (FB-R) as the primary signalling partner, driving pro-inflammatory stromal activation marked by upregulation of Ccl2, Ccl7, and Csf2. Intra-macrophage subcluster communication also intensified at this timepoint (447 interactions). These findings delineate the temporal and functional heterogeneity of cardiac macrophages during EAM progression and identify key immune–stromal interactions driving pathological cardiac remodelling. The coexistence of pro-inflammatory and transitional reparative macrophage subsets highlights the limitations of broad immunosuppression and supports precision strategies targeting CCR2-mediated recruitment, the SPP1 signalling axis, and macrophage–fibroblast crosstalk as therapeutic avenues in myocarditis and its progression.

Accurate taxonomic classification is essential for effective conservation strategies, especially for rare and endangered species. Traditionally based on morphology, such classifications may be challenged by recent advances in molecular tools that reveal hidden genetic structure or lack thereof. Jacobaea erucifolia is a declining species in Poland, primarily threatened by habitat loss, fragmentation, and competition with invasive species. Although two subspecies—erucifolia and tenuifolia—have been identified in Poland based on leaf morphology and ecological preferences, the taxonomic status of these forms remains unclear. This study aimed to evaluate the genetic variation within ten Polish populations of J. erucifolia and assess whether the observed morphological differences correspond to genetic divergence. Based on morphology, four populations were classified as subsp. erucifolia and six as subsp. tenuifolia. Reference populations of confirmed subspecies tenuifolia were included from Slovakia, Hungary, and the Netherlands. Genetic analyses were conducted using amplified fragment length polymorphisms (AFLPs), chloroplast DNA restriction fragments (RFLP), cpDNA, and nuclear internal transcribed spacer (ITS) ribosomal DNA sequences. The results revealed a high level of admixture and no significant genetic differentiation among Polish populations, regardless of their initial morphological classification. All Polish populations formed a single genetic group, distinct from foreign tenuifolia samples, although no diagnostic genetic markers were identified to separate these two groups (subspecies) of J. erucifolia definitively. Moreover, sequence data showed no differences among all analyzed individuals, further challenging the validity of subspecies differentiation. These findings suggest that the morphological variation observed within Polish populations likely reflects phenotypic plasticity rather than subspecies-level divergence. Consequently, subspecies tenuifolia does not appear to occur in Poland, and observed differences between Polish populations and those from neighboring countries may represent ecotypic variation. Our study underscores the necessity of integrating genetic and morphological data when resolving taxonomic ambiguities, especially in the context of conservation planning. Future research involving broader geographic sampling and deeper analysis of hybridization patterns will help clarify the evolutionary history and and population dynamics of J. erucifolia.

There is a high demand for the development of new carriers for pharmaceutical forms for human, veterinary, and animal-feeding use. One of the solutions might be bioactive compound-loading pellets for animal forage. The aim of the work was to assess the physical and sensory properties of forage with the addition of onion peel and off-spec onions as a source of quercetin. The feed was prepared using an expanding process (thermal–mechanical expanding process). Quercetin content was evaluated in raw onion and in final-product feed mixture samples (before and after expanding, and pelleting). The obtained feed was subjected to sensory analysis, testing for expanded pellet uniformity, water absorption index (WAI), the angle of a slide, and antioxidant activity. The results confirmed a high recovery of the quercetin after the expanding process (approximately 80%), and a significantly reduced intensity of onion odor, which was confirmed compared to the non-expanded onion, which is beneficial. Furthermore, digital and optical microscopy were applied for structure analysis. Microscopic imaging results confirmed that the onion structures were visible in the whole length of feed material and analyzed cross-sections. The results can be an introduction to further research on developing products that use the expanding and pelleting process to exploit the peel and off-spec onions, as well as other waste raw materials.

Introduction and objective: Mobile phones and Wi-Fi are the most commonly used forms of telecommunications. Initiated with the first generation, the mobile telephony is currently in its fifth generation without being screened extensively for any biological effects that it may have on humans or...

Plant cannabinoids, secondary metabolites of species belonging to the Cannabis genus, can mimic the endocannabinoids’ action and exert biological effects. Considering the contribution of the endocannabinoid system in cell cycle and apoptotic regulation, there is an interest in exploring the potential anti-cancer activities of natural and synthetic cannabinoids. Cannabidiol (CBD), an abundant plant cannabinoid, reveals a low affinity to cannabinoid receptors and, contrary to various cannabinoids, lacks psychoactive action. Here, we present the in vitro assessment of the pro-apoptototic potential of CBD-rich extracts of Cannabis sativa L. (eCBD) compared to purified CBD (pCBD). As demonstrated, both eCBD and pCBD decreased the viability of breast cancer cell line MDA-MB-231 and human prostate cancer cell line PC-3 in a concentration-dependent fashion. Endoplasmic reticulum stress-related apoptosis and morphological changes were induced only in low-serum conditions. Moreover, the effects of eCDB and pCDB were also assessed in non-malignant cell lines (MCF-10A and PNT2) with no alterations of viability noted, ultimately suggesting a selective action of CBD in tumor cells. The results suggest the possible involvement of reactive oxygen species in the response mechanism to eCBD and pCBD, but no clear pattern was observed. We also demonstrated significant changes in gene expression involved in apoptosis and cell cycle control upon extract treatment. Altogether, our study shows the potential of eCBD and pCBD as novel pro-apoptototic agents that can be considered promising in future preclinical and clinical testing.

The bacteria-derived CRISPR/Cas (an acronym for regularly interspaced short palindromic repeats/CRISPR-associated protein) system is currently the most widely used, versatile, and convenient tool for genome engineering. CRISPR/Cas-based technologies have been applied to disease modeling, gene therapies, transcriptional modulation, and diagnostics. Nevertheless, some challenges remain, such as the risk of immunological reactions or off-target effects. To overcome these problems, many new methods and CRISPR/Cas-based tools have been developed. In this review, we describe the current classification of CRISPR systems and new precise genome-editing technologies, summarize the latest applications of this technique in several fields of research, and, finally, discuss CRISPR/Cas system limitations, ethical issues, and challenges.

Progressive changes in the environment are related to modifications of the habitat. Introducing exotic species, and interbreeding between species can lead to processes that in the case of rare species or small populations threatens their integrity. Given the declining trends of many populations due to increased hybridization, early recognition of hybrids becomes important in conservation management. Natural hybridization is prevalent in Jacobaea. There are many naturally occurring interspecific hybrids in this genus, including those between Jacobaea vulgaris and its relatives. Although Jacobaea erucifolia and J. vulgaris often co‐occur and are considered closely related, apart from the few reports of German botanists on the existence of such hybrids, there is no information on research confirming hybridization between them. Morphologically intermediate individuals, found in the sympatric distributions of J. vulgaris and J. erucifolia, were hypothesized to be their hybrids. Two molecular marker systems (nuclear and chloroplast DNA markers) were employed to test this hypothesis and characterize putative hybrids. Nuclear and chloroplast DNA sequencing results and taxon‐specific amplified fragment length polymorphism (AFLP) fragment distribution analysis confirmed the hybrid nature of all 25 putative hybrids. The AFLP patterns of most hybrids demonstrated a closer relationship to J. erucifolia, suggesting frequent backcrossing. Moreover, they showed that several individuals previously described as pure were probably also of hybrid origin, backcrosses to J. erucifolia and J. vulgaris. This study provides the first molecular confirmation that natural hybrids between J. vulgaris and J. erucifolia occur in Poland. Hybridization appeared to be bidirectional but asymmetrical with J. vulgaris as the usual maternal parent.