Osteoporosis is closely linked to oxidative stress and inflammation, positioning the vitamin E metabolite γ-CEHC, known for its robust antioxidant and anti-inflammatory properties, as a promising therapeutic agent. However, its molecular targets have remained largely unknown. In this study, we characterized the protein targets of γ-CEHC and clarified its role in regulating bone metabolism using an ovariectomized (OVX) mouse model and invitro assays. Bone morphological analysis and histomorphometry demonstrated that γ-CEHC improves osteoporosis in OVX mice by inhibiting osteoclast differentiation and enhancing osteoblast differentiation. To identify the underlying mechanisms, we employed isothermal thermal proteome profiling (TPP) to map γ-CEHC-interacting proteins, followed by Gene Ontology (GO) and KEGG enrichment analyses. Our findings identified fatty acid-binding protein 5 (Fabp5) as a core target. The direct and specific binding between γ-CEHC and Fabp5 was confirmed through cellular thermal shift assays (CETSA), molecular docking-suggesting hydrogen bonding with Thr63-and Surface Plasmon Resonance (SPR) which showed a strong binding affinity (Kd = 5.24 μM). Furthermore, γ-CEHC was found to suppress LPS-induced M1 macrophage activation and promote M2 polarization, thereby reducing reactive oxygen species (ROS) levels and restoring bone remodeling homeostasis. This study is the first to systematically elucidate the molecular mechanisms of γ-CEHC in bone metabolism, revealing that it acts as a highly selective ligand for Fabp5. These findings provide a novel mechanistic basis for using γ-CEHC and targeting Fabp5 in the treatment of osteoporosis.

Dextromethorphan (DXM), a widely used antitussive agent, was investigated for its effects on mitochondrial F1FO-ATPase activity and oxidative phosphorylation. Our results demonstrate that DXM inhibited F1FO-ATPase independently of the thiol redox state. Mutual exclusion analysis highlighted an overlapping binding site between DXM and dicyclohexylcarbodiimide (DCCD), indicating a shared or adjacent binding site in the membrane-embedded FO domain of the enzyme. These findings suggested that DXM selectively targeted the proton translocation mechanism of F1FO-ATPase during the ATP hydrolysis and synthesis of ATP. Moreover, kinetic analysis confirmed a high affinity of DXM for the enzyme, with an inhibitory efficiency of 2.37 mM-1⸱s-1. Importantly, DXM did not affect electron transport chain activity but impaired ATP synthesis, as evidenced by altered respiratory control ratios of oxidative phosphorylation. The data obtained offer new insights into its off-target mitochondrial effects and potential implications for bioenergetic regulation.

Plant-based food products have been developed from diverse plant sources as new food choices. Fermentation of plant matrices with lactic acid bacteria (LAB) has been shown to improve quality and bioactivity of the resulting product while proteolysis by the LAB in the plant-based matrix remains to be elucidated. In this study, a hazelnut-based matrix prepared for a plant-based product was fermented with four different starter cultures of LAB, and their effects on proteolysis, bioactivity and allergenicity were investigated. Sucrose supplementation of the hazelnut matrix stimulated fermentation and time to reach to the target pH of 4.5 was shortened. CH-1 was the fastest acidifying culture reducing pH to the target value after 5 h. While the cultures RSF-736 and CHN-11 required 18 h for fermentation, R-707 was co-cultured with CH-1 to reach the target pH within the same time. Bacterial counts were in the range of 5-8 log cfu/g without a significant change after 15 days of storage in the hazelnut-based products. Level of proteolysis as measured by changes in soluble protein and total free amino acid contents differed among the cultures. Reductions in the amounts of hazelnut proteins were also confirmed by SDS-PAGE analysis, especially in the products prepared with cultures R-707+CH-1 and RSF-736. Allergenicity of the hazelnut matrix, determined by a hazelnut-specific ELISA test, significantly decreased after fermentation with all the cultures. Fermentation also enhanced total phenolic content and antioxidant activity of the hazelnut matrix with CHN-11 demonstrating the highest values after storage. On the other hand, fermentation did not significantly alter α-amylase inhibitory activity compared to the activity of 10.2% in the unfermented control. In addition, fermentation resulted in no change or a slight reduction in ACE inhibitory activity compared to the activity of 46.9% in the unfermented control depending on the culture. These findings demonstrate that LAB species can degrade hazelnut matrix leading to a reduction in allergenicity and enhancement of antioxidant activity.

Cutaneous squamous cell carcinoma (CSCC) is a malignant tumor originating from epidermal keratinocytes. In various types of tumors, ferroptosis is a vital iron-dependent form of regulated cell death. Recent studies suggest that heat shock protein 27 (HSP27), encoded by the heat shock protein family B member 1 (HSPB1) gene, is involved in the regulation of ferroptosis, but the specific mechanism remains unclear. In this study, CSCC cell lines were transfected with lentivirus-mediated HSPB1-shRNA or lentivirus carrying overexpressed HSPB1. CSCC cell lines, xenograft mouse models, and ferroptosis inhibitors or inducers were applied to verify the mechanism and function of HSP27. Downregulation of HSP27 inhibited the proliferation, migration, and invasion of CSCC cells, whereas upregulation of HSP27 showed the opposite results. Similarly, tumor volume and weight were reduced after HSP27 was downregulated invivo. Further studies revealed that HSP27 promoted the growth of CSCC cells and tumors by inhibiting ferroptosis, and the downregulation of HSP27 enhanced ferroptosis induced by Erastin. Ferrostatin-1 or Erastin successfully reversed the phenotype triggered by HSP27 alterations. HSP27 can induce the growth of CSCC by inhibiting ferroptosis, and is expected to become a new target for the treatment of CSCC.

Pancreatic cancer (PC) remains a highly lethal malignancy with limited treatment options, largely due to its heterogeneity and therapy resistance. While ferroptosis-a form of iron-dependent cell death driven by lipid peroxidation-has emerged as a relevant pathway, its role in PC is incompletely understood, as is the oncogenic function of Centrosomal Protein 55 (CEP55). Here, we integrated TCGA data with immunohistochemical validation and demonstrated that CEP55 is significantly overexpressed in PC and correlates with advanced disease and poor prognosis. Functionally, CEP55 knockdown suppressed proliferation, migration, and clonogenicity, while inducing ferroptosis, as evidenced by elevated lipid peroxidation, iron accumulation, and glutathione depletion. Mechanistically, CEP55 silencing downregulated key ferroptosis suppressors, including GPX4, SLC7A11, and NQO1, increasing cellular sensitivity to ferroptotic stress. Erastin, a ferroptosis inducer, enhanced ferroptosis in CEP55-deficient cells and counteracted the tumor-promoting effects of CEP55 overexpression. Invivo, CEP55 silencing reduced tumor growth and altered ferroptosis markers. Our findings establish CEP55 as a novel driver of PC progression via ferroptosis suppression, supporting its potential as both a prognostic biomarker and a therapeutic target for combination strategies aimed at overcoming PC resistance.

Trifuhalol A (TFA), a phlorotannin derived from the edible brown seaweed Agarum cribrosum, has been reported to exert diverse physiological activities, yet its anti-diabetic mechanism remains unclear. This study systematically investigates the multi-targeted anti-diabetic effects of TFA, with a particular focus on enhancing glucose uptake and protecting pancreatic islets. Invitro enzyme inhibition assays demonstrated that TFA significantly inhibited the activities of α-glucosidase and α-amylase, indicating its potential to attenuate postprandial glycemic excursions by modulating carbohydrate hydrolysis. Additionally, TFA effectively suppressed the formation of advanced glycation end-products (AGEs), potentially reducing the risk of diabetes-associated complications. Mechanistically, TFA enhanced glucose uptake in C2C12 myotubes by activating the PI3K/Akt and AMPK signaling pathways, which in turn promoted the translocation of glucose transporter type 4 (GLUT4) to the plasma membrane, thereby facilitating cellular glucose utilization and insulin sensitivity. Furthermore, invivo investigations using an alloxan-induced type 1 diabetic zebrafish further confirmed the bioefficacy of TFA, as evidenced by its capacity to reduce hyperglycemia, alleviate oxidative stress, and protect pancreatic islets, without eliciting observable systemic toxicity. Taken together, these findings provide both mechanistic and functional evidence supporting TFA as a safe and potent multi-target bioactive compound with promising applications in the development of functional foods and therapeutic strategies for diabetes management.

Exosomes are nanoscale extracellular vesicles (EVs) that have recently garnered significant attention owing to their crucial role in orchestrating cell-to-cell communication. Through the transfer of heterogeneous molecular cargo encompassing lipids, proteins, cytokines, growth factors, and RNAs (including mRNAs, lncRNAs, miRNAs, and circRNAs), they modulate a wide spectrum of physiological and pathological processes. Exosomes have been extensively investigated as diagnostic tools, therapeutic agents, as well as innovative platforms for drug delivery in metabolic, oncological, cardiovascular, and neurological disorders. Culminating evidence has demonstrated the pivotal role of exosomes in renal pathophysiology. Depending on their cargo content, exosomes represent potential biomarkers for early disease detection and survival prediction across various renal pathologies. While current therapeutic interventions are largely confined to attenuating disease progression, exosomes hold the potential to promote regeneration in both acute kidney injury and chronic kidney diseases. The current review comprehensively examines the clinical utility of exosomal cargo as diagnostic and prognostic biomarkers as well as therapeutic agents in kidney diseases, highlighting their crosstalk with critical signaling pathways implicated in renal pathophysiology. Addressing the current challenges in exosome isolation and standardization, and the development of advanced exosome engineering technologies are crucial for the transformation from experimental research settings to clinical practice. This should be augmented by preclinical validation and well-designed clinical trials, ultimately paving the way for a new era of precision medicine.

Glioblastoma (GB) is highly malignant with a median survival of 14 months despite conventional treatments like surgery, radiotherapy, and temozolomide. Resistance to these therapies necessitates innovative approaches, such as immune checkpoint inhibitors (ICIs) targeting cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), programmed cell death protein 1 (PD-1), and programmed death ligand 1 (PD-L1) to enhance T-cell-mediated tumor destruction. However, clinical trials have shown limited ICI efficacy in GB due to its immunosuppressive microenvironment and the blood-brain tumor barrier (BBTB), which impairs drug delivery. Emerging evidence highlights the gut microbiota as a pivotal modulator of ICI response, enhancing CD8+ and CD4+ T-cell function, antigen presentation, and immune modulation via the gut-brain axis in cancers. In addition, studies showed that gut-derived metabolites, including short-chain fatty acids, modulate immune responses and support blood-brain barrier integrity by regulating inflammatory signaling and tight junction proteins. Future GB research should prioritize clinical trials, mechanistic studies, and interventional strategies like fecal microbiota transplantation and probiotics to enhance ICI efficacy.

ABSTRACTMetabolic diseases have increased worldwide in recent decades, mainly due to a sedentary lifestyle and an unhealthy diet, with diet identified as an important regulator of gut microbiota composition. The use of natural products, such as Crocus sativus tepals extract (CTE) could be a promising approach to alleviate metabolic disorders. The aim was to investigate the potential ameliorative mechanisms of CTE in metabolic disorders induced by a high‐fat diet in an animal model, focusing on the composition of the gut microbiota and its relationship with the gut‐liver axis. We analyzed liver‐related biochemical and morphological parameters in mice fed a 60% fat diet for 14 weeks and orally treated with CTE during the last 5 weeks of the diet. In addition, jejunal and liver histology, intestinal barrier integrity, inflammation and oxidative stress, liver inflammation and lipid metabolism were investigated. The results showed that oral administration of CTE restored the composition of the gut microbiota and specifically promoted short‐chain fatty acids‐producing and anti‐inflammatory bacterial genera. It also improved intestinal barrier integrity and reduced inflammation in the jejunum and liver, along with a suppression of Fas and CerS6 expression in the liver and a reduction in circulating free fatty acids and β‐hydroxybutyrate levels. Our results indicate a possible link between the gut microbiota and the metabolic benefits of treatment with CTE, suggesting its therapeutic potential for the prevention or treatment of metabolic disorders.

Tumor suppressor Phosphatase and Tensin Homolog Deleted on Chromosome TEN (PTEN) shows a differential sub-cellular distribution, with its nuclear presence being particularly critical for its multifaceted tumor-suppressive functions. Nuclear PTEN mediates its arsenal of tumor suppressive actions viz., genomic stability maintenance, cell cycle regulation, DNA damage response, and transcriptional modulation, in both phosphatase-dependent and non-phosphatase-dependent manners. Diverse mechanisms exist to facilitate its nuclear import, including passive diffusion, active transport, and post-translational modifications such as monoubiquitination, phosphorylation, and SUMOylation as well as their crosstalk. Similarly, a number of mechanisms dictate the nuclear export of PTEN. Nucleo-cytoplasmic shuttling of PTEN is closely guarded by several protein factors. This review comprehensively explores the proteins involved in the transport and regulation of nuclear PTEN. Furthermore, it highlights the clinical significance of nuclear PTEN levels, which are closely associated with tumor grade, disease prognosis, and patient survival across multiple cancer types. By elucidating these mechanisms, this review underscores the importance of nuclear PTEN in cancer biology and its potential as a therapeutic target.