Search Results (21,636)

Mycobacterium tuberculosis (M. tuberculosis) has developed some strategies to evade host immune responses through ubiquitination, thereby facilitating persistent mycobacterial infection. The Rv3717 protein has been identified as a peptidoglycan (PG) amidase that contributes to mycobacterial survival, but its exact mechanism is still unclear. The findings of this study indicate that Rv3717 inhibits mycobacterial clearance within pulmonary epithelial cells. To elucidate the molecular mechanisms by which Rv3717 facilitates persistent infection, we identified intracellular candidates interacting with Rv3717 using co-immunoprecipitation (Co-IP) combined with liquid chromatography–mass spectrometry (LC-MS/MS). The unique proteins are categorized into three functional networks: mRNA splicing, the immune system process, and the translation process through Protein–Protein Interaction (PPI) analysis. The candidate interacting proteins of Rv3717 are involved in interleukin-2 enhancer-binding factor 2 (ILF2) and TAF15, as well as the polyubiquitin chain (UBC) and E3 ubiquitin ligase TRIM21. Our results suggest that intracellular Rv3717 is likely to influence biological processes through the potential interacting proteins. Our findings confirmed that Rv3717 interacted with interleukin enhancer-binding factor 2 (ILF2) through Co-IP and immunofluorescence assays. Furthermore, Rv3717 was verified to bind with ubiquitin and be degraded through the proteasome system. More importantly, the ubiquitination of Rv3717 accelerated the proteasomal degradation of ILF2 and downregulated the expression of IL-2. This study is the first to propose that the ubiquitination of the mycobacterial membrane vesicle-associated protein Rv3717 facilitates the proteasomal degradation of ILF2, resulting in the downregulation of IL-2 expression. Overall, the role of intracellular Rv3717 in promoting mycobacterial survival is associated with its ubiquitination and the proteasomal degradation of ILF2. Full article

Background: Although Ti-Fu-Kang (TFK) decoction has been clinically used for fatigue management, the systematic understanding of its mechanisms, particularly against central fatigue, remains largely unknown. This study is the first to employ an integrative approach of network pharmacology and metabolomics to explore the mechanisms of TFK against central fatigue. Methods: The central fatigue rat model was established using the modified multiple platform method in conjunction with alternate-day fasting. Behavioral alterations were evaluated through six behavioral tests, while brain injury was assessed through HE and Nissl staining. Serum metabolic indicators were analyzed to identify fatigue-related metabolic disturbances. Western blot analysis was used to assess the protein phosphorylation level of PI3K and AKT1. Oxidative stress was assessed by measuring superoxide dismutase, malondialdehyde, and glutathione peroxidase activities. Network pharmacology and serum metabolomics investigated the molecular mechanisms and metabolic pathways. Results: TFK significantly ameliorated behavioral abnormalities and brain pathological damage in central fatigue model rats. Network pharmacology analysis and in vivo experiment revealed that TFK may mediate biological processes such as oxidative stress and neuron death via the PI3K-AKT signaling pathway. Moreover, analysis of serum fatigue-related metabolic indicators indicated that TFK significantly modulated metabolic disruptions by elevating the levels of glucose, liver glycogen, and muscle glycogen and reducing the levels of alanine aminotransferase, aspartate aminotransferase, blood urea nitrogen, creatine kinase, lactate, and lactate dehydrogenase in central fatigue rats. Serum metabolomics analysis revealed that TFK ameliorates central fatigue by modulating amino acid metabolism, specifically by altering the levels of leucine and L-tryptophan, which subsequently contributes to the restoration of 5-hydroxytryptamine and dopamine homeostasis. Conclusions: This study elucidates the potential therapeutic mechanism of TKF in alleviating central fatigue, providing a scientific and theoretical basis for broader application and development of TFK. Full article

Ticks transmit diseases and harm animals worldwide, and their control primarily relies on pesticides. Resistance to these pesticides has developed consistently over centuries. Arginine Kinase (AK, EC 2.7.3.3) is a conserved, ancestral enzyme that provides reserve energy in emergency situations and a viable target for novel antiparasitic drugs. Our aim was to evaluate six carbamoyl carboxylic acid analogues (CCAs) as potential lead compounds by investigating their interaction with the active site of Rhipicephalus sanguineus AK (RsAK) using a structural modeling approach. The binding was characterized using fluorescence quenching (Stern–Volmer analysis) and molecular dynamics simulations. The simulations, performed with GROMACS using the CHARMM 26 force field over 100 ns, provided atomic-level insight into the ligand–protein interactions and stability. CCA4 exhibited the lowest dissociation constant (K_D~13·10⁻⁶ M) among the analogues, which we attribute to its end moieties (carboxylate and a pyridine on the ends). Purely aromatic ends (CCA1) or those with dual carboxylates (CCA6) showed lower affinity, suggesting that electrostatic complementarity and steric fit are processes involved in the binding. Despite requiring optimization, the CCA scaffold represents a novel strategy for tick control. These compounds provide a foundation for developing synergistic agents to enhance the efficacy of sustainable acaricides. Full article

The conventional kinesin (kinesin-1) molecular motor is a prototypical member of the kinesin superfamily. It can processively step on microtubules toward the plus end by hydrolyzing ATP molecules, performing the biological function of shuttling cargos in cells. Its dynamics have been thoroughly studied using various methods including biochemical measurement, single molecule imaging, single molecule optical trapping, and so on. While most of the experiments yielded consistent results on the dynamics of the motor, a lot of conflicting experimental results have also been presented. Here, a brief review is given of the diverse conflicting experimental results. Furthermore, a model for the chemomechanical coupling of the motor is briefly reviewed, which can consistently and quantitatively explain these conflicting experimental results in addition to the other experimental results. A consistent explanation of the diverse conflicting experimental results with the same model is an essential criterion for determining the correctness of the model. Full article

Cardiovascular aging is a multifactorial and systemic process that contributes significantly to the global burden of cardiovascular disease, particularly in older populations. This review explores the molecular and cellular mechanisms underlying cardiovascular remodeling in age-related conditions such as hypertension, atrial fibrillation, atherosclerosis, and heart failure. Central to this process are chronic low-grade inflammation (inflammaging), oxidative stress, cellular senescence, and maladaptive extracellular matrix remodeling. These hallmarks of aging interact to impair endothelial function, promote fibrosis, and compromise cardiac and vascular integrity. Key molecular pathways—including the renin–angiotensin–aldosterone system, NF-κB, NLRP3 inflammasome, IL-6, and TGF-β signaling—contribute to the transdifferentiation of vascular cells, immune dysregulation, and progressive tissue stiffening. We also highlight the role of the senescence-associated secretory phenotype and mitochondrial dysfunction in perpetuating inflammatory and fibrotic cascades. Emerging molecular therapies offer promising strategies to reverse or halt maladaptive remodeling. These include senescence-targeting agents (senolytics), Nrf2 activators, RNA-based drugs, and ECM-modulating compounds such as MMP inhibitors. Additionally, statins and anti-inflammatory biologics (e.g., IL-1β inhibitors) exhibit pleiotropic effects that extend beyond traditional risk factor control. Understanding the molecular basis of remodeling is essential for guiding future research and improving outcomes in older adults at risk of CVD. Full article

Seaweeds are increasingly valued in the food industry for their bioactive compounds, mainly hydrocolloids like carrageenan. This study investigates E. perplexum, a red seaweed, as a unique and sustainable source of carrageenan with promising functional properties. Using response surface methodology (RSM), the extraction process was optimized through alkaline extraction, identifying optimal conditions of 85 °C for 3 h with 2.58 M KOH, which yielded 77.10% carrageenan. The extracted carrageenan exhibited strong emulsifying activity (71.53 ± 2.41) and color properties comparable to commercial carrageenan, highlighting its viability for food applications. Chemical evaluation revealed a higher sulfate content (8.45 ± 0.16) and slightly reduced carbohydrate levels, which may influence its gelling and stabilizing abilities. Structural examination through ATR-FTIR spectroscopy corroborates the presence of key functional groups, including sulfate esters and galactose derivatives, inferring molecular integrity. These results emphasize the importance of RSM in optimizing extraction and underscore the ability of E. perplexum as a promising source of the derived carrageenan, which is a high-performance additive in food systems. Further research on purification, functional enhancement, and safety assessment is recommended to facilitate its integration into commercial food systems. Full article

Peptide mapping is a well-established method for confirming the identity of therapeutic proteins as part of batch release testing and product characterization for regulatory filings. Traditionally based on enzymatic digestion followed by reversed-phase liquid chromatography and UV detection, the method has evolved with technological advancements to incorporate mass spectrometry (MS), enabling more detailed structural insights. Residue-level confirmation of amino acid sequences requires MS/MS fragmentation, which produces large amounts of data that must be processed using specialized software. In regulated environments, the use of academic algorithms is often limited by validation requirements, making it necessary to rely on commercially approved tools, although their built-in scoring systems have limitations that can affect sequence assignment accuracy. Here, we present representative examples of incorrect peptide assignments generated by commercial software. In antibody sequence analysis, misidentifications resulted from isobaric and near-isobaric dipeptides (e.g., SA vs. GT). Additional examples from the analysis of SARS-CoV-2 spike protein variants revealed software-induced artifacts, including artificial succinylation of aspartic acid residues to compensate for sequence mismatches, and incorrect deamidation site assignments due to misinterpretation of isotopic peaks. These findings underscore the necessity for expert manual review of MS/MS data, even when using validated commercial platforms, and highlight the molecular challenges in distinguishing true sequence variants from software-driven artifacts. Full article

The global rise in the elderly population inherently escalates the demand for health and social care. Ensuring cognitive performance for healthy brain aging presents significant challenges for researchers and health professionals promoting self-care behaviors. This article aims to provide a comprehensive and critical analysis of the latest research on healthy brain aging by employing a biopsychosocial framework. It integrates biological, psychological, and social dimensions to elucidate their collective influence on cognitive health in older adults. Methodologically, this article provides a narrative review of the existing literature. A diverse array of bibliographic resources was obtained from prominent electronic databases, including MEDLINE, PubMed, Scopus and Web of Science, to ensure broad coverage of the topic. The search was designed to capture relevant studies published between 2010 and 2025, using key terms such as ‘aging’, ‘biomarker’, ‘neurodegeneration’, and ‘cognitive performance’. Following a rigorous selection process, two field specialists evaluated a total of 106 full-text articles to identify those that met the eligibility criteria, ultimately yielding 70 relevant studies. The findings reveal important connections between psychosocial and biological biomarkers and brain morphology, highlighting lifestyle factors—such as diet, exercise, and social engagement—as crucial for cognitive health. The article also underscores specific biomarkers relevant for assessing brain age and their relationship to neurodegenerative disorders. Notably, while biological markers like Aβ, tau, and α-synuclein (proteins that define the core molecular pathology of common neurodegenerative diseases) are present, they do not guarantee the onset of neurodegenerative diseases; psychosocial factors play an essential role in determining disease manifestation. In conclusion, these results support a holistic approach to healthy aging, which integrates psychosocial environments and lifestyle choices that enhance cognitive resilience. We propose further cross-sectional descriptive studies to better identify the biopsychosocial variables influencing cognitive performance and healthy brain aging, aiming to improve clinical practices and inform public health strategies. Full article

A reassessment of the risk-benefit balance of the two lipid nanoparticle (LNP)-based vaccines, Pfizer’s Comirnaty and Moderna’s Spikevax, is currently underway. While the FDA has approved updated products, their administration is recommended only for individuals aged 65 years or older and for those aged 6 months or older who have at least one underlying medical condition associated with an increased risk of severe COVID-19. Among other factors, this change in guidelines reflect an expanded spectrum and increased incidence of adverse events (AEs) and complications relative to other vaccines. Although severe AEs are relatively rare (occurring in < 0.5%) in vaccinated individuals, the sheer scale of global vaccination has resulted in millions of vaccine injuries, rendering post-vaccination syndrome (PVS) both clinically significant and scientifically intriguing. Nevertheless, the cellular and molecular mechanisms of these AEs are poorly understood. To better understand the phenomenon and to identify research needs, this review aims to highlight some theoretically plausible connections between the manifestations of PVS and some unique structural properties of mRNA-LNPs. The latter include (i) ribosomal synthesis of the antigenic spike protein (SP) without natural control over mRNA translation, diversifying antigen processing and presentation; (ii) stabilization of the mRNA by multiple chemical modification, abnormally increasing translation efficiency and frameshift mutation risk; (iii) encoding for SP, a protein with multiple toxic effects; (iv) promotion of innate immune activation and mRNA transfection in off-target tissues by the LNP, leading to systemic inflammation with autoimmune phenomena; (v) short post-reconstitution stability of vaccine nanoparticles contributing to whole-body distribution and mRNA transfection; (vi) immune reactivity and immunogenicity of PEG on the LNP surface increasing the risk of complement activation with LNP disintegration and anaphylaxis; (vii) GC enrichment and double proline modifications stabilize SP mRNA and prefusion SP, respectively; and (viii) contaminations with plasmid DNA and other organic and inorganic elements entailing toxicity with cancer risk. The collateral immune anomalies considered are innate immune activation, T-cell- and antibody-mediated cytotoxicities, dissemination of pseudo virus-like hybrid exosomes, somatic hypermutation, insertion mutagenesis, frameshift mutation, and reverse transcription. Lessons from mRNA-LNP vaccine-associated AEs may guide strategies for the prediction, prevention, and treatment of AEs, while informing the design of safer next-generation mRNA vaccines and therapeutics. Full article

Constructed wetlands (CWs) are sustainable and cost-effective systems that utilise plant–microbe interactions and natural processes for wastewater treatment. Microbial communities play a pivotal role in pollutant removal by crucial processes like nitrogen transformations, phosphorus cycling, organic matter degradation and the breakdown of emerging contaminants. Dominant phyla, such as Proteobacteria, Bacteroidetes, Actinobacteria and Firmicutes, collectively orchestrate these biogeochemical functions. Advances in molecular tools, including high-throughput sequencing and metagenomics, have revealed the diversity and functional potential of wetland microbiomes, while environmental factors, i.e., temperature, pH and hydraulic retention time, strongly influence their performance. Phosphorus removal efficiency is often lower than nitrogen, and large land requirements and long start-up times restrict broader application. Microplastic accumulation, the spread of antibiotic resistance genes and greenhouse gas emissions (methane, nitrous oxide) present additional challenges. The possible persistence of pathogenic microbes further complicates system safety. Future research should integrate engineered substrates, biochar amendments, optimised plant–microbe interactions and hybrid CW designs to enhance treatment performance and resilience in the era of climate change. By acknowledging the potential and constraints, CWs can be further developed as next-generation, nature-based solutions for sustainable water management in the years to come. Full article

Background/Objectives: Transient neonatal diabetes mellitus (TNDM) is a form of neonatal diabetes mellitus (NDM) arising in the first weeks of life and remitting in infancy. Epigenetic aberrations at the imprinted 6q24 locus (overexpression of PLAGL1/HYMAI) are the most common causes of TNDM. The aim of this study was a retrospective clinical and genetic analysis of a Polish pediatric cohort, emphasizing the role of methylation-specific MLPA (MS-MLPA) in the diagnosis of TNDM. Methods: We conducted a retrospective analysis of the medical records of 22 patients with diabetes diagnosed at 1 year of age. The molecular studies included an analysis of the NDM gene panel by a targeted NGS and MS-MLPA for the 6q24 imprinting region. Results: 6q24-TNDM was confirmed in five patients, with a median age of diabetes remission of 4 months (IQR: 3–6 months). The MS-MLPA identified paternal UPD6 or isolated maternal hypomethylation of PLAGL1 in three patients, and two had a paternal 6q24 duplication. Conclusions: In our group, changes in the 6q24 region were confirmed in 22.7% of NDM patients, indicating the usefulness of the MS-MLPA technique in the diagnosis and detection of imprinting defects. We acknowledge key limitations, including diagnostic delays and incomplete parental testing, which precluded trio-based confirmation of paternal UPD6 versus epimutation in some cases; future diagnostic workflows should incorporate an early trio-based SNP array or STR confirmation. A methylation analysis should be included early in the NDM genetic diagnosis process to provide genetic counseling and monitor patients for diabetes recurrence. Full article

Background: Low temperature stress is a major environmental challenge affecting the growth, metabolism, and survival of many aquaculture species, including Nile tilapia (Oreochromis niloticus). Understanding the molecular mechanisms underlying cold tolerance is therefore essential for improving fish resilience and aquaculture sustainability. Methods: In the present study, an acute cold stress model of Nile tilapia (Oreochromis niloticus) was established and it was found that uncoupling protein 1 (UCP1) was involved in the acute cold stress process of tilapia. Results: The upregulation of UCP1 in the liver under cold stimulation was regulated by stress hormones such as cortisol and adrenaline. UCP1 has a short half-life and is degraded by proteasomes. In tilapia primary hepatocytes, the addition of adrenergic receptor agonists resulted in mitochondrial membrane potential decreasing, while UCP1 siRNA transfection inhibited mitochondrial membrane potential. Biochemical characteristics indicate that UCP1 is a channel protein that mediates proton leakage. In addition, feeding and intraperitoneal injection of mitochondrial uncoupling agent BAM15 can alleviate the low-temperature stress of tilapia. Conclusions: UCP1 helps maintain the metabolic homeostasis of tilapia under acute cold stimulation and provides new insights into the mechanisms of cold resistance as well as potential treatment strategies in fish. Full article

Cholangiocarcinoma (CCA) is an aggressive malignancy of the biliary tract with rising global incidence and limited treatment options. Its pathogenesis involves a complex interplay of genetic mutations, epigenetic dysregulation, inflammatory signaling, and environmental influences. Emerging evidence highlights the pivotal role of the gut–liver axis and microbiota dysbiosis in shaping biliary homeostasis and disease progression. Alterations in microbial composition disrupt apoptosis and autophagy, two key processes regulating cell survival and death, thereby contributing to tumorigenesis, metastasis, and therapy resistance. Specific taxa, including Enterococcus, Escherichia coli, Pseudomonas, Bifidobacterium, and Bacillus, demonstrate strain-dependent effects, acting either as tumor promoters through genotoxic metabolites and immune evasion or as potential tumor suppressors by inducing apoptosis and immune activation. These findings underscore the context-dependent roles of microbiota in CCA biology. Importantly, microbiota modulation offers novel therapeutic opportunities. Dietary interventions such as probiotics, prebiotics, and nutritional strategies, alongside innovative microbiome-targeted therapies, hold promise for restoring microbial balance, enhancing antitumor immunity, and improving patient outcomes. This review integrates current molecular and microbiological evidence to propose the gut microbiota as both a biomarker and a therapeutic target in CCA, opening avenues for precision medicine approaches in hepatobiliary oncology. Full article

Refrigeration has become a common practice for preserving Dendrobium officinale products. The molecular mechanisms underlying chilling stress responses, particularly those linking physiological adaptation to flavor-related metabolite changes, remain unclear. This study aimed to explore the transcriptional and metabolic changes in D. officinale leaves during cold treatment and to identify key stress-responsive metabolites underlying flavor modulation and their roles in cold adaptation. Transcriptional clustering analysis revealed distinct expression profiles under varying temperatures, indicating that chilling temperatures affect pathways related to RNA processing, oxidative stress, and secondary metabolism. Metabolomics profiling demonstrated significant metabolite shifts over time, with lipids, organic acids, and phenylpropanoids being prominently altered. Notably, flavonoids like rutin and sugars like trehalose varied in their accumulation depending on the duration of cold exposure. Proteomic analysis indicated that proteins involved in amino acid metabolism and the TCA (tricarboxylic acid) cycle were significantly impacted by prolonged chilling, with amino acids (key osmoprotectants and flavor contributors) accumulating over time, linking cold stress adaptation to sensory quality enhancement. These findings suggest that a chilling temperature primarily affects metabolic flow at different time points, which could help control the quality of D. officinale leaves during cold storage. Full article

Circadian rhythms are endogenous ~24 h oscillations that regulate diverse biochemical processes. Although stress responses can exhibit circadian modulation, evidence for rhythmic regulation of stress granules (SGs)—cytoplasmic RNA–protein condensates formed under stress—remains limited. We investigated sodium arsenite-induced SG dynamics in NIH/3T3 cultures. SG number, eIF3 signal intensity—an established SG marker—and area oscillated with a period of ~24 h. These rhythms persisted in Bmal1^−/− mouse embryonic fibroblasts (MEFs), despite lacking a transcription–translation feedback loop (TTFL) that constitutes the canonical circadian clock, but with altered amplitude and phase, indicating partial dependence on the molecular clock. Several SG-associated RNA-binding proteins (TIA-1, BRF1, hnRNP Q, and LARK) exhibited time-dependent changes at the mRNA and/or protein level, suggesting potential mechanisms for rhythmic SG modulation. Unlike previous in vivo reports linking SG variation to eIF2α phosphorylation, no temporal changes in phosphorylated eIF2α were observed, highlighting differences between isolated cells and tissues. Our results show that SG rhythmicity can persist without BMAL1, supporting alternative oscillatory mechanisms that contribute to the temporal organization of stress responses. Given their role in cell survival and the association of SG dysfunction with disease, these rhythms provide insight into how cellular stress responses are temporally regulated. Full article