Search Results (13,653)

Background/Objectives: Variants of MYPN, encoding a sarcomeric protein myopalladin, are associated with different types of cardiomyopathies and myopathies. However, the molecular mechanisms of MYPN-associated pathologies are still poorly understood. Methods: In this study, we generated induced pluripotent stem cells (iPSCs) from a hypertrophic cardiomyopathy patient carrying a novel p.N989I (c.2966A>T) variant of MYPN and used iPSC-derived cardiomyocytes to examine the impact of the variant on biophysical characteristics and transcriptomic profile. Results: No significant changes in parameters of calcium transients, sodium current and action potential were found in iPSC-derived cardiomyocytes with the p.N989I (c.2966A>T) variant of MYPN compared to non-isogenic cells from an unrelated healthy donor. At the transcriptomic level, MYPN-N989I cardiomyocytes demonstrated an upregulation of genes linked to cell cycle, mitotic spindle, microtubule cytoskeleton organization, and myogenic program genes. Downregulation of sarcomeric, Z-disc- and cell junction-associated genes, as well as genes involved in ATP synthesis, oxidative phosphorylation, and the SRF-signaling pathway, was also revealed. Conclusions: Our data suggest that the p.N989I (c.2966A>T) variant of MYPN plays a dual role in hypertrophic cardiomyopathy pathogenesis, disrupting not only sarcomeric and cytoskeletal organization but also the regulation of the muscle gene program. Full article

The European Union aims to achieve climate neutrality by 2050, with biogas and biomethane expected to play an increasingly important role in the decarbonisation of the energy system. This study investigates the economic and social determinants shaping the development of biogas production in European countries and identifies an optimal investment strategy for new biogas plants under varying environmental conditions. An expert–mathematical method was applied to assess and hierarchise twenty economic and social factors influencing biogas production, based on evaluations provided by 71 experts from eleven European countries. Subsequently, individual choice criteria derived from game theory were used to determine the optimal strategy for biogas plant construction under conditions of uncertainty. The results indicate that six determinants—EU-level production support mechanisms, investment costs, national support instruments, process efficiency improvements, community involvement, and agricultural raw material prices—account for 52.9% of the total impact on biogas development potential. Among the analysed investment options, large-scale biogas plants with an installed capacity of 3 MW were identified as the optimal strategy, offering the lowest unit production costs and the lowest risk of cost overruns across diverse economic and social environments. These findings provide policy-relevant insights for supporting efficient and socially acceptable biogas deployment in Europe. Full article

Aging is a regulated process marked by the accumulation of senescent cells, which remain viable but no longer divide. Senescent cells contribute to age-associated phenotypes and diseases, including osteoarthritis, dementia, and cancer, but their scarcity and heterogeneity have limited study. Here, we developed a fluorescence-guided high-resolution spatial transcriptomic profiling approach to precisely locate and profile p21-reporter-positive cells in aged liver. This method enabled unbiased detection of a p21-associated, senescence-enriched cell population and revealed its diverse cellular identities, including hepatocytes, macrophages, neutrophils, and plasma cells. Our analysis further showed that activated macrophages and hepatic stellate cells were more likely to exhibit a p21 positive (p21⁺) state than their resting counterparts. Transcriptomic profiling of p21-expressing cells indicated heterogeneous senescence-associated secretory phenotype (SASP) programs, with distinct inflammatory and remodeling signatures across cell identities and their spatial positions. In parallel, we identified an aggregation of interferon-stimulated gene (ISG)-expressing cells with limited overlap with p21 positivity, suggesting a distinct aging-associated stress program. Taken together, our fluorescence-guided spatial transcriptomic framework enables high-resolution, single-cell mapping of senescence in situ, delineating both senescent cell type specificity and cell identity–independent senescence programs, thereby advancing a more comprehensive understanding of regulatory mechanisms underlying aging. Full article

To date, the problem of mastitis in cattle remains relevant for both the industrial sector and scientific research. Despite numerous active investigations, the causes of this disease have not been fully established. It is postulated that several factors may be involved, such as bacterial pathogens, animal husbandry practices, and weather and climatic conditions. In this study, we selected cows from farms in Yakutia to investigate microbial isolates present in the milk of cows affected by mastitis and treated with antibiotics. Five identified Staphylococcus aureus isolates were investigated using whole-genome sequencing (Illumina sequencing and nanopore sequencing), followed by analysis of virulence factors in the genomes and cultural properties of the isolates. The profile of S. aureus virulence genes (exotoxins, cytotoxins, superantigen-like proteins, adhesins) was identified via WGS. Hemolysin gene (hla) was detected in all isolates. An investigation of the cultural properties of the isolates, specifically through hemolysis of rabbit erythrocytes and Western blot analysis of the culture liquid of S. aureus, revealed different expression levels of alpha-hemolysin among the strains. One isolate (17-21) exhibited the highest secretion level of about 320 ± 37 ng, both in the hemolysis test and immunoblotting assay. An investigation of the isolates’ antibiotic resistance showed that all isolates exhibited multidrug resistance, as confirmed by the presence of antibiotic resistance genes in these isolates. One isolate (7-7) exhibited the broadest range of phenotypic resistance and was resistant to all tested antibiotics (except clindamycin). Phylogenetic analysis suggested that the evolution of these isolates occurred independently in their respective ecological niches, although their transfer from cattle to humans, and vice versa, is possible. Isolates 7-7, 18-22, 33-40, and 35-42 are most typical to Yakutian cattle, while isolate 17-21 might have been introduced from a different region. To the best of our knowledge, this is the first in-depth study into a range of S. aureus isolates associated with mastitis infection in Yakutian cattle. Full article

Cardiovascular diseases remain a leading cause of morbidity and mortality worldwide, underscoring the need to better understand cardiovascular physiology. A key aspect involves identifying regulatory molecules that govern metabolic shifts. Forkhead box protein O1 (FoxO1) has emerged as a potential regulator; however, its role and underlying mechanisms remain unclear. This study investigated FoxO1 in metabolic adaptation using Wistar rats divided into age groups (fetal, postnatal day 1, postnatal day 7, adult) and treatment groups (control, hypoxia, FoxO1 inhibitor, combination). Hypoxia (12–14% O₂) and FoxO1 inhibitor (AS1842856, 10 mg/kgBW/day) were administered accordingly. Parameters assessed included hypoxia inducible factor 1 α (HIF-1α), FoxO1 mRNA and protein, glucose transporter type 1 (GLUT1), glucose transporter type 4 (GLUT4), cluster of differentiation 36 (CD36), hexokinase, pyruvate dehydrogenase kinase isoform 4 (PDK4), phosphoenolpyruvate carboxykinase (PEPCK), lactic acid, malonyl-CoA, carnitine palmitoyltransferase 1 (CPT1), citrate synthase, cytochrome c, and adenosine triphosphate (ATP). ATP production increased with age, associated with higher FoxO1 expression and metabolic shifts. Hypoxia in fetal hearts reduced HIF-1α and FoxO1. FoxO1 inhibition elevated glycolytic and oxidative markers. In conclusion, FoxO1 regulates glycolysis and lipid metabolism, offering insights into cardiac adaptation to hypoxia and potential therapeutic strategies. Full article

Estimation of the postmortem interval (PMI) remains a major challenge in forensic science. We used attenuated total reflection (ATR)–Fourier-transform infrared (FTIR) spectroscopy combined with chemometric modeling for PMI prediction using vitreous humor samples from 20 forensic cases with known PMI (24.8–97.6 h) and 10 with unknown PMI. The intensities of vibrational bands commonly associated with PMI were analyzed, and several peaks in the carbohydrate/phosphate region showed significant correlations with PMI. Principal component analysis revealed time-dependent spectral evolution, with PC1 (48.1%) associated mainly with carbohydrate/phosphate variations and PC2 (37.6%) with protein structural changes. Partial least squares regression with two latent variables achieved a cross-validated RMSE of 15.8 h (R² = 0.53) on all 20 known samples. Variable importance analysis identified glycoprotein degradation (1190 cm⁻¹) and phospholipid breakdown (736 cm⁻¹) as the dominant predictors, with traditional carbohydrate bands playing a secondary role. Predictions for unknown samples ranged from 27.1 to 80.1 h, with five of ten falling within the 90% prediction interval (±20 h) of the available estimates. This study presents a promising PMI estimation model that performed well on unseen samples, even if the sample size represents a methodological limitation that will be addressed in future investigations through larger, more diverse datasets. Full article

Peptide deformylase (PDF) belongs to a conserved enzyme family critical for N-terminal methionine excision (NME), an essential protein maturation process in prokaryotes and eukaryotic organelles (chloroplasts, mitochondria). To explore the potential functions of OsPDFs in Oryza sativa, this study employed bioinformatics approaches and experimental validation to systematically identify and analyze the OsPDF gene family. Three OsPDF genes (OsPDF1A, OsPDF1B, OsPDF1B2) were identified in rice. These genes are exclusively distributed on chromosome 1. The biophysical properties of these proteins showed that OsPDF1A and OsPDF1B are alkaline proteins, while OsPDF1B2 is acidic, and all are hydrophilic with moderate thermostability potential. Synteny analysis revealed closer evolutionary relationships between Oryza sativa and the monocot Triticum aestivum than with dicots, reflecting conserved PDF function in gramineous plants. Analysis of cis-acting elements in the 2000 bp upstream region of OsPDF gene promoters revealed numerous elements associated with abiotic stress response and hormone regulation. Furthermore, quantitative real-time PCR (qRT-PCR) data supported these findings, indicating that OsPDF1A and OsPDF1B were upregulated under low-temperature stress, and all three OsPDF genes were transcriptionally activated by heat, salt and UV-B stresses, indicating their active involvement in rice growth, development, and abiotic stress tolerance. In summary, OsPDFs exhibit significant functions in rice’s stress adaptation, growth, and development. This study not only enhances our understanding of the OsPDF gene family’s genomic, evolutionary, and functional characteristics, but also provides new perspectives and foundational data for further exploring their regulatory mechanisms in protein maturation and abiotic stress responses, as well as their potential applications in rice stress tolerance breeding. Full article

Although LRRK2 mutations modulate systemic glucose homeostasis and metabolic dysfunction precedes Parkinson’s disease (PD) motor symptoms; the way in which pathogenic variants of LRRK2 disrupt astrocytic glucose metabolism and organellar homeostasis remains poorly understood. Here, we demonstrate that LRRK2-I1371V mutation causes profound metabolic and organellar dysfunction in LRRK2-I1371V PD-iPSC-derived astrocytes and U87 cells overexpressing I1371V variant. LRRK2-I1371V astrocytes exhibit significantly reduced GLUT1 expression and cell surface localization, resulting in impaired glucose uptake and decreased lactate production. This metabolic insufficiency correlates with cascading mitochondrial dysfunction, characterized by membrane depolarization, elevated reactive oxygen species, enhanced ubiquitination and reduced proteasomal activity. Reduced LAMP1/LAMP2 expression, impaired lysosomal acidification, and selective cathepsin D deficiency were observed. Accumulation of undegraded cargo was confirmed by transmission electron microscopy upon α-synuclein exposure. ER stress was evident by upregulation of GADD34/CHOP, increased phospho-PERK, and reduced nascent protein synthesis. Increased ER–mitochondrial contact via MAMs and enhanced STIM1-ORAI3 clustering reflect compensatory but ultimately insufficient responses to energy stress. Our results reveal that LRRK2-I1371V induces glucose uptake deficits, leading to energy depletion and integrated ER–mitochondria–lysosome dysfunction, thus indicating restoration of astrocytic metabolic capacity as a potential therapeutic strategy for LRRK2-associated PD. Full article

Parameters of reflected light, measured in narrow or broad spectral bands, are widely analyzed for remote and proximal sensing of plant responses to stressors. Specifically, parameters of reflectance in red (R), green (G), and blue (B) spectral bands measured using simple color images can be sensitive to characteristics of plants. The conventional view is that RGB reflectance primarily reveals long-term changes in plants (days, weeks, etc.). In this study, we investigated light-induced changes in RGB reflectance in wheat (Triticum aestivum L.) and pea (Pisum sativum L.) leaves. Illumination increased this reflectance for about 10 min in wheat and about 15–20 min in pea; these changes relaxed after light intensity was decreased. The changes in RGB reflectance were strongly related to the effective quantum yield of photosystem II and non-photochemical quenching of chlorophyll fluorescence under high light intensity; these relations were absent under low light intensity. We hypothesized that changes in both RGB reflectance and photosynthetic parameters were related to the light-induced changes in chloroplast localization. A simple mathematical model of optical properties and photosynthesis in leaves was developed; results of the model-based analysis supported the proposed hypothesis. Experimental analysis of the dynamics of light transmittance additionally supported this hypothesis. Our results thus show that RGB imaging can be sensitive to fast changes in plants. Full article

Two-pore channels (TPCs) are evolutionarily conserved intracellular cation channels found in both plants and animals, where they mediate ion fluxes across endomembrane compartments. While historically the plant channel was among the first plant ion channels to be characterized, thanks to the relative ease of applying the patch-clamp technique to isolated plant vacuoles, where it is localized, the functional properties of the two main human isoforms, HsTPC1 and HsTPC2, expressed in endosomal and lysosomal membranes, were elucidated much later. In plants, TPCs are typically represented by a single isoform, exemplified by AtTPC1 in the model plant Arabidopsis thaliana, which functions as a voltage-dependent, Ca²⁺-regulated channel. The physiological role of plant TPCs is not yet fully clarified, although evidence suggests that they may contribute to systemic signaling and stress responses. In humans, two main isoforms, HsTPC1 and HsTPC2, are expressed in endosomal and lysosomal membranes. Human TPCs are primarily regulated by the phosphoinositide PI(3,5)P₂ and display a high selectivity for Na⁺. However, these channels also appear as a non-selective cationic conductance when activated by the potent Ca²⁺-mobilizing messenger NAADP, likely through interaction with an accessory protein. Functionally, human TPCs are involved in endolysosomal trafficking, membrane fusion, and intracellular signaling, with emerging roles in immunity, metabolism, and disease. Overall, TPCs represent key components of intracellular ion homeostasis and cellular physiology; however, their precise regulatory mechanisms and integrated physiological roles remain only partially understood and, in several respects, are still elusive. Full article

Background: The aim of this study is to investigate the relationship between the expression levels of autophagy-related genes (SQSTM1, Beclin1, Atg5, and Atg7) in diffuse astrocytic tumors and clinicopathological parameters, including tumor grade, IDH mutation status, and survival outcomes. Materials and Methods: A total of 150 histopathologically confirmed diffuse astrocytic tumor cases were retrospectively analyzed. Clinical data were extracted from patient records. Gene expression levels were determined using qRT-PCR and evaluated by the 2^−ΔCt method, where lower ΔCt values indicate higher gene expression. IDH1 R132H mutation status was evaluated by immunohistochemistry. Results: No statistically significant differences were observed in the expression levels of SQSTM1, Beclin1, Atg5, and Atg7 across WHO tumor grades (p > 0.05). However, when analyzed by IDH status, IDH-mutant tumors exhibited significantly higher gene expression levels (demonstrated by lower ΔCt values) of Beclin1 (p = 0.046) and Atg5 (p = 0.027) compared to IDH wild-type tumors. In multivariate Cox regression analysis, age and WHO tumor grades were confirmed as independent prognostic factors. Crucially, higher SQSTM1 expression independently predicted worse clinical outcomes, specifically poorer overall survival (OS) (p = 0.004) and shorter progression-free survival (PFS) (p = 0.031). Additionally, elevated Beclin1 expression was identified as an independent predictor of worse OS (p = 0.023). Conclusions: This study demonstrates that increased expression of autophagy-related genes, particularly SQSTM1 and Beclin1, serves as a robust indicator of poor prognosis and shorter survival times in diffuse astrocytic tumors. Furthermore, the elevated expression of Beclin1 and Atg5 in IDH-mutant cases highlights a complex metabolic interplay that warrants further investigation as potential therapeutic targets. Full article

Thermoplastic polyurethane properties are governed by the interplay between soft-segment mobility, hard-segment interactions, and segmented morphology, yet the extent to which atomistic predictions of their thermal and mechanical behavior depend on force-field choice remains insufficiently benchmarked. Here, we combine FTIR, DSC, TGA, and tensile testing with all-atom molecular dynamics simulations to investigate HDI–PEG polyurethane systems across a controlled soft-segment series. Experimentally, films with PEG molecular weights of 400, 1000, and 1500 g/mol were characterized, while simulations were extended to 400–2000 g/mol to compare two complementary force-field frameworks under a consistent protocol: OPLS-AA, a conventional atom-type-based force field, and OpenFF/Sage, a direct-chemical-perception framework augmented here with bespoke torsional refinements. Both force fields reproduce the composition-driven decrease in Tg and density with increasing PEG length, but differ systematically in absolute values, with OPLS-AA predicting higher densities and Tg values than OpenFF. Tensile experiments show the highest elastic modulus for PEG400, a marked decrease at PEG1000, and a partial recovery at PEG1500. Although nanosecond-scale deformation simulations overestimate absolute moduli because they probe high-rate elastic response, they recover composition-dependent stiffness differences, with OpenFF yielding a more pronounced non-monotonic trend than OPLS-AA. Overall, this work provides an experimentally anchored benchmark for assessing which composition-driven trends in HDI–PEG polyurethanes are robust across force-field families, and which observables remain sensitive to model assumptions and simulation scale. Full article

Ionizing radiation affects enzymes, which are essential for most cellular functions, by inducing chemical alterations in their molecular structures, often resulting in the inhibition of their activities. Unraveling the molecular and kinetic mechanisms driving these effects requires irradiation protocols that ensure accurate dose delivery, spatial homogeneity, and reproducibility. In this study, we established a systematic experimental framework that adapts a medical linear accelerator (LINAC) as a precision source for biochemical irradiation experiments. A rigorous protocol was developed that allows enzyme solutions to be irradiated under strictly defined and verifiable dosimetric conditions. Using this approach, we quantified the radiation-induced modulation of enzyme activity in two representative enzymes: invertase (β-fructofuranosidase) and collagenase. For invertase, a pronounced nonlinear decrease in enzyme activity was observed, with the enzyme retaining approximately only 2.2% of its initial activity at 50 Gy. Conversely, collagenase activity exhibited an exponential dose–response behavior over the dose range 0.1–200 Gy, yielding a global inactivation constant of $K = 0.015 {\mathrm{G}\mathrm{y}}^{-1}$. Complementary SDS–PAGE analysis revealed no detectable radiation-induced protein fragmentation or aggregation under the investigated conditions. These results confirm enzyme-specific radiation sensitivity and demonstrate the efficacy of this LINAC-based methodology for quantitative dose–effect studies. Overall, this work provides a versatile experimental tool for applied radiation research, bridging the gap between clinical medical physics and fundamental biochemistry. Full article

The present study proposes a fractal-inspired multiscale framework to interpret the structural, thermal, mechanical and dielectric properties of polylactic acid (PLA). Experimental investigations were performed using tensile testing, TG-DTA thermal analysis, X-ray diffraction (XRD) and dielectric spectroscopy. The structural organization was analyzed using XRD data, where a scaling tendency compatible with power-law behavior was identified over a limited $\mathrm{q}$-range. The thermal degradation exhibited a sharp transition, while the mechanical and dielectric responses reflected the heterogenous behavior typical of semicrystalline polymers. Rather than claiming a fully validated fractal model, the present work introduces a conceptual multiscale interpretation, supported by experimental observations, and proposes a fractal integrity index (FII) as an exploratory descriptor integrating structural, thermal and mechanical information. The results suggest that fractal-based descriptors may provide a useful complementary framework for interpreting complex polymer behavior, although further validation across multiple materials and experimental conditions is required. Full article

Weak alternating electric fields are widely used in neuromodulation techniques such as transcranial alternating current stimulation (tACS), yet the precise biophysical mechanisms underlying neuronal responses remain incompletely understood. Current computational models often neglect the electrical properties of the extracellular microenvironment, limiting their predictive accuracy. Motivated by experimentally observed frequency-dependent modulation of neuronal activity, we developed a two-compartment model of hippocampal CA3 pyramidal neurons in which extracellular resistance is explicitly parameterized and systematically examined as a key factor influencing neuronal response properties under external electric fields. Within a dual-compartment Hodgkin–Huxley framework, the neuron is divided into a “soma–basal dendrite unit” and an “apical dendrite unit,” accounting for voltage polarization induced by external fields. Using phase-locking ratio curves and three-dimensional parameter response surface, we systematically characterized neuronal sensitivity to field parameters and examined how potassium equilibrium potential ($V_K$) and extracellular resistance ($R_{\mathrm{out}}$) modulate these responses. Our results demonstrate that increasing $R_{\mathrm{out}}$ enhances neuronal responsiveness to external fields, while $V_K$ variations primarily regulate intrinsic excitability. These findings provide mechanistic insights into the frequency-dependent modulation of neuronal responses under weak electric fields, consistent with phenomena observed in biological neural systems, and provide a mechanistic and theoretical framework for understanding the joint effects of electric field amplitude and frequency on neuronal sensitivity to weak electric fields, which may help inform future neuromodulation strategies. Full article