Search Results (1,966)

Cancer cells are able to survive under conditions of high endoplasmic reticulum (ER) stress by activating the adaptive unfolded protein response (UPR), which is closely linked with autophagy. On the other hand, excessive and prolonged ER stress leads to apoptosis. However, the relationships between different UPR branches and apoptosis or autophagy vary in cancer cells of different origins and depend on the extent and nature of the stress signal. This study was designed to establish the role of ER stress sensors protein kinase RNA-like endoplasmic reticulum kinase (PERK) and inositol-requiring enzyme 1 (IRE1α) in apoptosis or autophagy signaling in T-lymphoblastic leukemia MOLT-3 cells via the RNA interference method. The cells were transfected with small interfering RNAs (si-PERK, si-IRE1α, or si-Cont) for 6 h and further cultured under normal conditions for 72 h to provide an insight into chronic effects of the gene silencing. The expression of apoptosis and autophagy effectors at the mRNA and protein levels was compared using RT-PCR and Western blot assays, respectively. Transfection of the cells with PERK siRNA led to a significant decrease in PERK protein and gene expression, and decreased phosphorylation of its downstream effector eukaryotic initiation factor 2α (eIF2α). PERK silencing was accompanied by activation of apoptosis-related genes and proteins—BCL2-associated X (Bax), caspase-3, C/EBP homologous protein (CHOP), while the levels of autophagy markers (Unc-51 like autophagy activating kinase 1 (ULK1), Beclin-1, and microtubule-associated proteins 1A/1B light chain 3 (LC3A/B)) remained stable. In contrast, treatment of the cells with si-IRE1α reduced the content of IRE1α, X-box-binding protein 1 (sXBP1), and glucose-regulated protein 78 (GRP78) proteins, but increased ERN1 gene expression. IRE1α RNA interference did not affect the levels of the pro-apoptotic marker Bax, but suppressed caspase-3, CHOP, c-Jun N-terminal kinase (JNK), and autophagy signaling molecules (ULK1, Beclin-1, LC3A/B) at both the transcriptional and translational levels. These results indicate that the PERK pathway is an important contributor to the survival of MOLT-3 cells under basal ER stress, while PERK depletion compromises the resistance of cells to UPR-mediated apoptosis. The IRE1α UPR branch is directly linked with autophagy-dependent signaling, although IRE1α knockdown exerted a more complicated influence on the cells, probably via activation of multiple pro-death and compensatory pro-survival regulatory mechanisms. Full article

Sarcopenia is an age-related progressive decline in skeletal muscle mass, strength, and physical performance that increases the risk of falls, disability, and reduced quality of life among older adults. Its pathogenesis is multifactorial and involves chronic low-grade inflammation, hormonal disturbances, insulin resistance, and mitochondrial dysfunction, leading to an imbalance between muscle protein synthesis and degradation. The aim of this study was to summarise current knowledge regarding the mechanisms underlying sarcopenia, contemporary diagnostic methods, and the effectiveness of modern nutritional and exercise-based strategies, with particular emphasis on technologies supporting patient monitoring. This study is a structured narrative review conducted across PubMed, Scopus, and Web of Science databases, with the literature search completed on 1 March 2026. Separate searches were performed for thematic sections, including pathophysiology, diagnosis, physical activity, nutritional interventions, plant-derived compounds, and digital health technologies. While the core search focused on publications from 2023–2025, specific time-bound deviations were applied: the search for plant-derived compounds was extended back to 2020, and combined interventions were searched up to March 2026 to ensure the inclusion of the most recent evidence. The review included 53 peer-reviewed primary studies (RCTs and observational) and secondary literature (reviews and meta-analyses) involving individuals aged ≥60 years. The most robust evidence supports multicomponent interventions, particularly the synergy between resistance training and adequate protein intake (1.2–1.5 g/kg/day), often supplemented with leucine, vitamin D, omega-3 fatty acids, and creatine. Such strategies effectively counteract anabolic resistance by combining mechanical loading with the stimulation of the mTORC1 signalling pathway, leading to significant improvements in muscle mass, strength, and physical function. While isolated protein or micronutrient supplementation shows limited effectiveness in the absence of exercise, their role as supportive elements in multimodal strategies is well-documented. Furthermore, emerging digital health technologies—including wearable sensors and telerehabilitation—are proving essential for clinical practice, enabling precise, continuous monitoring of physical activity and gait parameters under free-living conditions, which enhances both patient adherence and long-term therapeutic outcomes. Full article

Cerebellar climbing fiber–Purkinje cell (CF–PC) long-term depression (LTD) plays a critical role in motor learning and is modulated by locus coeruleus (LC) noradrenergic afferents via distinct adrenergic receptor (AR) subtypes. Nevertheless, the mechanisms underlying LC noradrenergic neuron-mediated regulation of CF–PC LTD remain poorly understood. Here, we investigated the effects of chemogenetic activation of LC noradrenergic afferents on CF–PC LTD in cerebellar slices from dopamine β-hydroxylase (DBH)-Cre mice using electrophysiology, glutamate sensor imaging, immunofluorescence and pharmacological approaches. Tetanic stimulation (5 Hz) of CFs induced CF–PC LTD under control conditions, and this LTD was enhanced by chemogenetic activation of LC noradrenergic afferents. Blockade of group I metabotropic glutamate receptors (mGluR1) abolished LTD under control conditions, whereas chemogenetic activation of LC noradrenergic afferents triggered a novel form of CF–PC LTD accompanied by an increased N2/N1 ratio. With mGluR1 blocked, chemogenetic activation of LC noradrenergic afferents failed to trigger the novel CF–PC LTD following blockade of α₂-AR or α₂A-AR, but not α₂B-AR or α₂C-AR. Importantly, chemogenetic activation of LC noradrenergic afferents triggered LTD of glutamate fluorescence at CF terminals, which was abolished by blockade of α_2-AR or α₂A-AR, but not α₂B-AR or α₂C-AR. Notably, inhibition of either cyclin-dependent kinase 5 (CDK5) or presynaptic, but not postsynaptic, protein kinase A (PKA) completely abolished the CF–PC LTD triggered by chemogenetic activation of LC noradrenergic afferents in mouse cerebellar slices. Immunofluorescence results showed robust α₂A-AR expression throughout the cerebellar molecular layer, with intense signals along PC dendrites and clear colocalization with vesicular glutamate transporter 2 (vGluT2) at cerebellar CF terminals. These results indicate that activation of LC noradrenergic afferents potentiates CF–PC LTD by triggering Glu-LTD at CF terminals through the α₂A-AR/CDK5/PKA signaling cascade in the mouse cerebellar cortex. Full article

Grass carp (Ctenopharyngodon idella) is an important freshwater aquaculture species in China. However, high-density farming often leads to significant growth differentiation among individuals, seriously affecting yield and product quality. Here, we conducted an integrated transcriptomic and data-independent acquisition (DIA) proteomic analysis across the brain, liver, and muscle tissues of fast-growing (FG) and slow-growing (SG) grass carp after 9 months of high-density culture. Our analysis revealed that the core mechanism driving growth differentiation is a deep decoupling of transcription and translation, where enhanced muscle translational efficiency dictates the fast-growth phenotype, while slow growth is constrained by central stress and hepatic energy depletion. In the slow growth group, the brain exhibited translational arrest and neuroinflammation, and the liver entered a state of hypermetabolism mediated by AMPK, evidenced by the post-transcriptional up-regulation of key sensors such as CAB39 (PRM ratio = 1.634) and CAMKK2 (PRM ratio = 1.295). Conversely, in the fast-growing group, the mTOR–ribosome signaling axis was strongly activated at the post-transcriptional level in the muscle (GSEA NES = −1.852), triggering myofibrillar protein deposition despite transcriptional silence in classical growth pathways. These findings elucidate the systemic molecular mechanisms of growth divergence and provide high-confidence molecular targets for developing fast-growing, stress-resilient grass carp strains for precision aquaculture breeding. Full article

Exosomes and small extracellular vesicles are promising liquid-biopsy biomarkers because they carry molecular information from their cells of origin and can be accessed from minimally invasive biofluids. Reliable separation and detection are made more difficult by their small size, low abundance, diverse composition, and co-occurrence with lipoproteins, protein aggregates, and other extracellular particles. To improve exosome enrichment, capture, and sensing, nanomaterial-assisted techniques have become crucial. Using a mechanism-based approach that differentiates between non-gravimetric signal amplification and genuine physical mass loading, this study offers an organized comparison of nanomaterial-enabled exosome sensing techniques. This distinction is helpful because different transducers measure different physical quantities: while optical, electrochemical, fluorescent, catalytic, and nucleic acid-based platforms typically benefit from enhanced signal generation rather than increased mass, resonant and gravimetric sensors benefit from increased inertial or surface-bound mass. In terms of amplification mechanism, transducer compatibility, sample-matrix tolerance, workflow complexity, and translational maturity, the review contrasts metallic nanoparticles, magnetic systems, metal–organic frameworks, carbon and two-dimensional materials, quantum dots, upconversion nanomaterials, DNA nanostructures, and polymer-based platforms. The gap between analytical sensitivity and clinical utility, including separation purity, recovery, biological heterogeneity, pre-analytical variability, interference from complex biofluids, and the need for uniform validation, is given special focus. The review concludes that no single nanomaterial or amplification method is universally optimal; instead, platform-aware, application-specific integration of isolation, amplification, and validation techniques is necessary for clinically meaningful exosome sensing. Full article

The detection of RASSF1A (Ras-associated domain family 1 isoform A) promoter methylation in body fluids can offer a powerful tool for the early diagnosis of bladder cancer. Zinc finger proteins (ZFPs) serve as sequence-specific recognition elements for targeting double-stranded DNA sequences. Here, we report a cyclic olefin copolymer (COC) chip-integrated ZFP array-based molecular sensor that bypasses the need for bisulfite conversion and PCR amplification to recognize the specific site of DNA methylation in the RASSF1A promoter. Building upon the SEER-LAC (SEquence-Enabled Reassembly of β-Lactamase) framework, we engineered a dual-recognition split-enzyme system in which a COC chip-immobilized ZFP array confers sequence specificity while a co-recruited methyl-binding domain (MBD) enforces methylation-dependent gating, together driving the proximity-induced reconstitution of functional β-lactamase at methylated target loci. Accordingly, this sensor specifically reassembles and restores enzymatic activity only in the presence of specific methylated DNA in the RASSF1A promoter region. We demonstrate that this dual-component array effectively differentiates methylation status with high specificity. Given its rapid turnaround and non-PCR-based mechanism, this system can be well-suited for developing diagnostic assays for bladder cancer, offering a potential alternative to conventional epigenetic screening methods. Full article

Calcineurin B-like (CBL) proteins are plant-specific calcium sensors critical for ion homeostasis and stress tolerance. Here, seven MdCBL genes were genome-wide identified in apple (Malus domestica). We conducted systematic bioinformatic profiling of their physicochemical features, subcellular localization, cis-regulatory elements, phylogeny, secondary structures, phosphorylation sites, and functional annotations and further verified the salt-stress regulatory function of MdCBL5 via transgenic tests. MdCBL proteins contain 210–246 amino acids, with molecular weights of 24,196.73–28,283.39 Da, pI values of 4.63–4.97 and instability indices of 37.12–49.20. Localization prediction placed these proteins in nuclei, cytosol and chloroplasts. Their promoter regions are rich in hormone-responsive (auxin, ABA, salicylic acid) and stress-responsive (cold, drought, salt) cis-elements. Phylogenetic clustering divided MdCBLs into five subgroups (A–E) with high homology to Arabidopsis CBLs. Random coils and α-helices dominate their secondary structures, and serine residues constitute most phosphorylation sites. Functional annotation supports their involvement in calcium signaling, ion transport and diverse stress adaptation. Salt stress experiments have confirmed that MdCBL5 may enhance apple salt resistance by promoting MdSOS gene expression. Meanwhile, transient transformation in apple fruit showed that MdCBL5 can effectively enhance fruit quality traits. Collectively, this study establishes a theoretical foundation for further elucidating the biological functions of the apple MdCBL5 gene and provides valuable insights for genetically improving stress resistance and fruit quality in apple via molecular breeding strategies. Full article

Salmonella is a globally significant foodborne intracellular pathogen, and invasive salmonellosis poses a major global public health threat. The NLR family CARD-containing protein 4 (NLRC4) inflammasome, a pivotal cytosolic innate immune sensor, specifically recognizes Salmonella flagellin and type III secretion system (T3SS) components via the NAIP (NLR family apoptosis inhibitory protein) family. Upon activation, it triggers pyroptosis, pro-inflammatory cytokine release, and infected intestinal epithelial cell extrusion, serving as a central pathway for host defense against Salmonella colonization and systemic spread. This work systematically summarizes the structural composition, activation mechanisms, post-translational modifications, and regulatory protein network of the NLRC4 inflammasome, and highlights the molecular mechanisms by which Salmonella evades NLRC4 surveillance through multiple strategies: transcriptional downregulation of immunogenic ligands, structural modification of T3SS components, secretion of effector proteins, and chemotaxis-virulence synergy. A comprehensive delineation of the co-evolutionary arms race between Salmonella and the NLRC4 inflammasome provides an integrated mechanistic framework for understanding host–pathogen immune interplay. Deciphering the mechanisms of bacterial immune evasion on this basis holds critical importance for identifying novel anti-infective targets and advancing translational preventive and therapeutic strategies against salmonellosis. Full article

Cyclic adenosine monophosphate (cAMP) is a ubiquitous second messenger downstream of G protein-coupled receptors (GPCRs) and a central regulator of cellular signaling. Genetically encoded exchange proteins directly activated by cAMP (Epac)-based Förster resonance energy transfer (FRET) biosensors enable real-time monitoring of cAMP dynamics in living cells, but commonly used cyan/yellow FRET pairs require short-wavelength excitation, limiting compatibility with multiplex imaging and blue-light optogenetic tools such as bacterial photoactivated adenylyl cyclases (bPACs). Here, we engineered and systematically characterized four red-shifted Epac-based single-chain FRET cAMP sensors combining yellow or orange FRET donors with red fluorescent FRET acceptors. Using ratiometric live-cell imaging, we quantified stimulus-evoked FRET responses and identified Epac_red4 as the best-performing variant, showing an approximately 55% decrease in normalized FRET after forskolin stimulation. Epac_red4 also reliably detected G_i/o-mediated decreases in cAMP following μ-opioid receptor activation. Brief 405 nm light pulses induced graded and reversible cAMP elevations using the low dark-activity variant bPAC-F198Y. Furthermore, Epac_red4 enabled analysis of cAMP recovery kinetics during phosphodiesterase inhibition and multiplex imaging of cAMP and intracellular Ca²⁺ using Fura-2 with minimal spectral and pH-related interference under physiological imaging conditions. Together, Epac_red4 represents a robust red-shifted cAMP sensor for optogenetic and multiplex signaling studies. Full article

Despite advances in liver machine perfusion (MP), ischemia–reperfusion injury (IRI) remains a major challenge in liver transplantation, with energetic stress and mitochondrial dysfunction recognized as key drivers of damage exacerbation. We investigated whether fractions enriched with extracellular vesicles (EVs) derived from mesenchymal stromal cells can preserve energetic homeostasis in rat livers undergoing normothermic MP (NMP). An established NMP rat model was used (n = 5 per group). After procurement, livers underwent NMP for 4 h, preceded or not by 30 min cold ischemia (CI). EVs (NMP + EVs and CI + NMP + EVs) or saline (NMP and CI + NMP) were randomly administered to the perfusion fluid. Perfusate samples were collected throughout the procedure, and biopsies were taken at the end of NMP. Ischemic livers exhibited succinate accumulation, flavin mononucleotide (FMN) release, activation of reverse electron transport, and adenosine triphosphate (ATP) depletion. EV treatment effectively counteracted these effects, restoring a metabolic profile comparable to that of non-ischemic livers. Moreover, EVs improved adenosine monophosphate/ATP ratios and prevented AMP-activated protein kinase activation, a key energy-stress sensor. Furthermore, EVs reduced oxidative stress markers, cell death mediators, and pro-inflammatory cytokines, indicating a broad cytoprotective and anti-inflammatory effect. These findings support the potential of EVs to preserve mitochondrial function, restore energy balance, and reduce inflammation, thereby improving liver cell viability during NMP. Full article

The small heat shock protein HSPB1 is a ubiquitously expressed mechanoresponsive chaperone essential for cytoskeletal remodeling under mechanical load. Mutations in HSPB1, including S135F, cause Charcot-Marie-Tooth (CMT) peripheral neuropathy, yet the mechanisms underlying the selective vulnerability of peripheral nerves remain enigmatic. Here we demonstrate that substrate stiffness is a critical determinant of HSPB1^S135F-mediated neurodegeneration. Using stiffness-tunable polydimethylsiloxane (PDMS) substrates (1 kPa, 10 kPa, 2 MPa) and uniaxial cyclic stretch, we show that primary dorsal root ganglia (DRG) neurons and SH-SY5Y cells expressing HSPB1^S135F exhibit profound deficits in mechanoadaptation. On compliant substrates (10 kPa), HSPB1^S135F causes stretch-induced axon fragmentation and neuronal death, whereas HSPB1^WT confers robust neuroprotection. HSPB1^S135F also disrupts stiffness-directed neuritogenesis in differentiated SH-SY5Y cells: HSPB1^WT-expressing cells show optimal axonal outgrowth and βIII-tubulin expression on 10 kPa substrates mimicking muscle tissue stiffness, while HSPB1^S135F mutants display disorganized focal adhesions and complete differentiation failure. Mechanistically, we uncover that HSPB1^S135F dysregulates stage-specific transglutaminase (TGase) expression—insufficient TGase during early neuritogenesis impairs filopodia stabilization, whereas aberrant TGase persistence at late stages constrains axon extension. Our findings establish HSPB1 as a biomechanical sensor that integrates ECM stiffness signals to coordinate peripheral nerve regeneration, and identify defective mechanoadaptation as a previously unrecognized pathomechanism in CMT. These results open new avenues for stiffness-targeted therapeutic strategies in peripheral neuropathy. Full article

Imiquimod-induced skin inflammation is the most widely used psoriasis mouse model. Although p38 mitogen-activated protein kinase reportedly plays a role in the pathogenesis of psoriatic inflammation, the purpose of one of its upstream activators, apoptosis signal-regulating kinase 1 (ASK1), remains unclear. This study investigated the role of ASK1 and its molecular mechanism in the imiquimod-induced psoriasis model. Compared to wild-type mice, the ASK1 knockout (KO) mouse skin lesion showed a higher clinical score and a thicker epidermis. The mRNA expression of pro-inflammatory cytokines, such as IL-17 and TNF-α, was also higher. Notably, the expression of aryl hydrocarbon receptor (AhR), a sensor for xenobiotic chemicals that is expressed in the skin to strengthen the skin barrier and accelerate terminal differentiation of the epidermis—as well as its downstream molecule CYP1A1, but not NRF2—was increased in the ASK1 KO psoriatic skin lesion. Immunoprecipitation analysis, followed by Western blotting, revealed that ASK1 interacts with AhR in cells transfected with their respective expression vectors, potentially leading to reduced AhR expression. These results suggest that ASK1 negatively regulates the development of the imiquimod-induced mouse psoriasis model by interacting with AhR and presumably antagonizing the AhR-CYP1A1 axis. Full article

As hen eggs are a primary source of high-quality dietary protein, egg freshness is fundamentally linked to biochemical alterations during storage, including moisture redistribution, protein degradation, and fluctuating chemical profiles. Accurate assessment of these internal changes is paramount for quality control; nonetheless, conventional analytical techniques remain predominantly destructive, rendering them impractical for high-throughput industrial monitoring. While existing literature has explored individual spectroscopic methods, the synergistic potential of multi-sensor integration and advanced artificial intelligence (AI) algorithms remains insufficiently synthesized. This review systematically evaluates recent breakthroughs in integrating AI with diverse spectroscopic modalities for non-destructive freshness quantification, including Visible-Near-Infrared (VIS-NIR), Raman, Fluorescence, and Hyperspectral Imaging (HSI). We elucidate the underlying mechanisms of spectral response to internal quality degradation and discuss the evolution of data-driven modeling from traditional chemometrics to sophisticated deep learning architectures. Furthermore, this work identifies critical bottlenecks in real-time industrial implementation and proposes future research trajectories toward intelligent multi-sensor fusion platforms. Full article

Food contamination by Aflatoxin M1 and Atrazine remains a critical food-safety concern, requiring sensitive detection methods that can operate reliably in complex matrices. Here, we report an AI-assisted antibody-functionalized electrochemical sensing platform for the detection and classification of Aflatoxin M1 and Atrazine across corn, corn flour, and protein matrices. The sensor used analyte-specific antibodies immobilized on an electrochemical electrode surface, where target binding produced measurable changes in the interfacial electrochemical response. Sensor performance was evaluated using cyclic voltammetry, coulometry, and electrochemical impedance spectroscopy (EIS), with EIS providing strong frequency-dependent signatures for concentration-dependent analysis. Spike-and-recovery studies further demonstrated the applicability of the platform in food-matrix conditions. To improve interpretation of complex electrochemical signals, full-spectrum EIS features were integrated with machine learning models for concentration-level classification into low, mid, and high groups. The AI workflow achieved an overall classification accuracy of 93.33%, with 96.67% specificity, 93.44% PPV, 96.66% NPV, and 0.982 AUC for Atrazine, and 96.70% specificity, 93.38% PPV, 96.67% NPV, and 0.987 AUC for Aflatoxin M1. In addition, analyte classification between Aflatoxin M1 and Atrazine reached 97.4% accuracy and 0.994 ROC-AUC. Overall, this work demonstrates a matrix-aware electrochemical immunosensing strategy enhanced by AI-based signal interpretation for food contaminant detection. Full article

The use of insect-based protein sources in aquaculture is gaining increasing attention with Hermetia illucens (black soldier fly, BSF) larvae meal representing a promising substitute to fishmeal (FM). This study evaluated the effect of partial dietary inclusion of BSF meal (BSF0, BSF2.5, BSF5, BSF10%) on the volatilome of rainbow trout (Oncorhynchus mykiss) fillets, before and after cooking, using gas chromatography–mass spectrometry (GC–MS) and a metal oxide sensor-(MOX)-based device. Fish were fed diets with increasing BSF inclusion, and both raw and cooked fillets were analyzed to assess changes in volatile organic compounds (VOCs). GC–MS enabled the identification and semi-quantitative analysis of VOC classes, while MOX sensor responses were processed using Linear Discriminant Analysis (LDA) to assess discrimination among dietary treatments. Results showed that BSF inclusion influenced the volatile profile, with clearer separation at higher inclusion levels (BSF5–BSF10%), especially in cooked fillets. Thermal processing enhanced these differences. GC–MS analysis revealed a reduction in aldehydes and ketones and an increase in carboxylic acids with higher BSF inclusion. Key compounds such as hexanal and heptanal decreased, indicating changes in lipid-derived volatile pathways. Overall, the integration of GC–MS and MOX sensors proved effective in detecting diet-induced changes, supporting their application as effective and reliable tools for quality assessment in aquaculture products, with potential implications for sensory quality that should be further confirmed through dedicated sensory studies. Full article