Type 2 diabetes (T2DM), a chronic metabolic disorder characterized by pancreatic β-cell dysfunction and insulin resistance, is closely associated with oxidative stress. Interferon-stimulated gene 15 (ISG15), a ubiquitin-like modifier, has been implicated in redox regulation; however, its mechanistic role in T2DM progression remains unclear. To identify novel bio-factors contributing to T2DM, peripheral blood mononuclear cells (PBMCs) and serum samples were collected from T2DM patients and age-/BMI-matched healthy controls to quantify ISG15 expression. Complementary studies utilized a T2DM mouse model to assess β-cell-specific ISG15 expression within pancreatic islets. In parallel, MIN6 pancreatic β-cells were exposed to pathophysiological stressors, including hyperglycemic/hyperlipidemic (HG/PG) conditions and hydrogen peroxide (H2O2)-induced oxidative stress, to mimic the diabetic microenvironment. Protein-protein interactions were evaluated, and functional analyses were conducted following ISG15 genetic ablation. The results demonstrated that ISG15 expression was significantly elevated in PBMCs and serum from T2DM patients compared to healthy controls. Similarly, T2DM mouse models exhibited marked upregulation of ISG15 in islet β-cells. In vitro, exposure to HG/PG or H2O2 increased ISG15 levels, reduced MIN6 cell viability, and heightened reactive oxygen species (ROS) accumulation. Mechanistic studies revealed that ISG15 directly interacts with the PI3K regulatory subunit PIK3R2, suppressing its activity and thereby disrupting the PI3K/AKT/Nrf2 antioxidant signaling axis. This disruption led to exacerbated oxidative stress. Collectively, these findings indicate that ISG15 acts as a novel mediator of oxidative stress in T2DM by targeting and inhibiting the PIK3R2-dependent PI3K/AKT/Nrf2 pathway. These results uncover a previously unrecognized molecular mechanism driving β-cell dysfunction and identify ISG15 as a potential therapeutic target for mitigating oxidative damage in diabetes.
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.
Hydroxy-safflor yellow A (HSYA), a bioactive compound from Carthamus tinctorius, has reported anti-inflammatory and antitumor activities. However, its effects on breast cancer and the underlying immune mechanisms remain poorly defined. In this study, we investigated the immunomodulatory role of HSYA in breast cancer, with a focus on its impact on tumor-associated macrophages (TAMs) and T cell responses. Using in vitro assays, murine breast cancer models, and single-cell transcriptomic analysis, we found that HSYA significantly inhibited tumor growth and reshaped the tumor immune microenvironment. Flow cytometry and transcriptomic profiling revealed that HSYA treatment reduced tumor-associated macrophages (TAMs) infiltration and suppressed C-C motif chemokine ligand 5 (CCL5) expression, which was associated with enhanced recruitment and activation of tissue-resident memory T (TRM) cells. Co-culture and functional assays further supported a role for the TAM/CCL5 axis in mediating these immune effects. Collectively, our findings demonstrate that HSYA exerts antitumor activity, at least in part, by modulating the TAM/CCL5/TRM axis, highlighting its potential as an immunomodulatory therapeutic strategy in breast cancer.
Glioma, a highly aggressive brain cancer, thrives in an immunosuppressive tumor microenvironment (TME). The Unfolded Protein Response (UPR) is a key adaptive stress pathway implicated in therapy resistance. To discover clinically actionable biomarkers, we employed an integrated multi-omics strategy, combining single-cell and bulk transcriptomic data from multiple glioma cohorts. Using hdWGCNA network analysis, we identified a UPR-associated gene module that defined two molecular subtypes with stark survival differences. Through a rigorous machine learning pipeline (Random Survival Forest, LASSO, and CoxBoost), ANXA5 emerged as the central prognostic gene. High ANXA5 expression consistently predicted poorer overall survival across eight independent datasets, validating its robust prognostic value. Functionally, ANXA5 was linked to extracellular matrix remodeling and immune modulation. Multi-omics profiling revealed that ANXA5-high gliomas exhibit a T-cell-inflamed yet immunosuppressive TME, characterized by elevated immune checkpoint expression. Crucially, ANXA5 demonstrated strong predictive power for response to immune checkpoint blockade (ICB), showing significant correlation with nine established immunotherapy response signatures and accurately discriminating responders from non-responders in six independent ICB-treated clinical cohorts (AUC: 0.65-0.78). Genomic analysis associated ANXA5 expression with distinct mutation patterns (EGFR/PTEN vs. IDH1/TP53). In vitro knockdown of ANXA5 confirmed its oncogenic role, as it suppressed glioma cell proliferation and invasion. Our study establishes ANXA5 as a prime example of a translatable biomarker discovered through multi-omics integration. It functions dually as a prognostic indicator and a predictive biomarker for immunotherapy, offering a tangible framework for patient stratification and personalized therapeutic strategies in glioma, thereby bridging a critical gap toward clinical translation.
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.
Fermented foods are increasingly recognized for their health-promoting properties, but little is known about the effects of sauerkraut brine (SB), a by-product of cabbage fermentation, on systemic inflammation and neuro-inflammatory responses. This study aimed to investigate the immunomodulatory, antioxidant, and behavioral effects of SB oral treatment in a mouse model of low-dose lipopolysaccharide (LPS) challenge. The SB was prepared by traditional spontaneous cabbage fermentation and analyzed for pH, microbiological profile, and acidity. At the end of fermentation, the brine contained a high level of viable lactic acid bacteria (LAB) (1.8 × 106 CFU/ml). Mice were pretreated orally with SB, heat-treated SB (htSB), or saline for 4 weeks before single or repeated LPS injection (0.5 mg/kg). Safety assessments, including body weight, food intake, and locomotor activity, did not indicate adverse effects with either form of SB. In the acute LPS model, SB pretreatment significantly reduced mRNA expression of the pro-inflammatory cytokines TNF-α, IL-1β, and IL-6 in the prefrontal cortex compared to the saline-pretreated group. Interestingly, the expression of IL-10, strongly induced by LPS, was also significantly reduced by SB, suggesting modulation of both pro- and anti-inflammatory signaling pathways. These protective effects were less pronounced in the heat-treated SB group, suggesting that viable bacteria or heat-sensitive components may be critical for bioactivity. In the repeated LPS model, SB prevented LPS-induced depletion of glutathione (GSH) and preserved total antioxidant capacity (TAC), while heat-treated SB provided no protection and resulted in increased protein carbonylation and decreased TAC. No significant changes in the activities of antioxidant enzymes (SOD1/2, CAT, GR, GSH-Px, GST) were observed. Behavioral tests using the open-field paradigm showed that all groups exhibited prolonged sickness behavior after repeated LPS exposure, despite a two-day washout period. However, only the SB group showed delayed recovery, with reduced horizontal and vertical activity lasting 1 day longer than in the other groups. Nevertheless, pretreatment with SB and heat-treated SB prevented the LPS-induced reduction in time spent in the central zone, a commonly used measure of anxiety-like behavior, suggesting a possible anxiolytic effect. In conclusion, SB exhibits anti-inflammatory and antioxidant properties associated with systemic LPS-induced neuroinflammation and may reduce anxiety-like behavior. These effects appear to be dependent on heat-sensitive constituents or microbial viability. The findings support further investigation of SB as a functional dietary intervention targeting neuro-immune health.
L-dopa decarboxylase (DDC) is the biosynthetic enzyme of dopamine and serotonin. Although DDC has been originally studied for its role in neurotransmission, it has also been detected in peripheral organs, where it is implicated in cellular homeostasis. DDC has been identified by our research team as a negative regulator of dengue virus (DENV) replication in liver cells. The latter has been attributed, at least in part, to the physical interaction of DDC protein with phosphatidylinositol 3-kinase (PI3K), and its biosynthetic function. PI3K/AKT signaling and cell survival are manipulated by DENV to favor its replication. Based on the above, we investigated whether DDC exerts its antiviral activity against DENV propagation through modulation of DENV-induced cell death and especially apoptosis. Specifically, DDC silencing in Huh7.5 cells (shDDC) significantly reduced virus-induced cytopathic effect compared to the control cells (shControl). This finding was accompanied by suppression of both early and later stages of apoptosis in the silenced cells, as shown by Annexin V/PI staining and TUNEL assay, respectively. Accordingly, upon infection, shDDC cells showed suppressed activation of key caspases, BCL-2 family members and TRAIL-receptor genes that modulate the apoptotic cascade, compared to the control cells. Moreover, mitochondrial analysis in DENV-infected cells revealed that, upon DDC silencing, a less pronounced disruption of mitochondrial membrane potential and network integrity, higher respiratory capacity, lower ROS production, and reduced cytochrome c release were observed. As the PI3K/AKT pathway is known to be affected by both DENV and DDC, next we assessed whether DDC is involved in the virus-induced apoptosis through this axis. For this, we quantified the reduction of p-AKT and p-mTOR levels caused by DENV infection in the two cell lines, which was found greater in the shControl cells. Finally, chemical inhibition of AKT phosphorylation abolished the differences in cell viability and apoptosis between the two cell lines. In total, our findings highlight the suppressive role of DDC against DENV replication by modulating the PI3K/AKT-dependent apoptotic signaling.
Resveratrol (RSV; 3,5,4'-trihydroxy-trans-stilbene) is a natural polyphenolic compound with notable antioxidant, anti-inflammatory, and immunomodulatory properties. It has been investigated for therapeutic applications in cardiovascular disease, cancer, and neurodegenerative disorders. This review emphasizes the potential of RSV in oncology and neuroprotection, synthesizing evidence from systematic database searches and experimental studies. Despite promising biological activities, RSV is limited by poor stability, low aqueous solubility, rapid metabolism, and restricted bioavailability, necessitating improved delivery strategies such as nanoencapsulation, nanocrystals, prodrugs, and structural analogues. Mechanistically, RSV exerts anticancer and neuroprotective effects through modulation of p53, STAT3, NF-κB, and mitochondrial-mediated apoptosis. Its antioxidant actions involve regulation of reactive oxygen species (ROS), activation of NRF2, AMPK signaling, and SIRT1. RSV and related antioxidants act on multiple molecular pathways, including TP53, β-catenin, STAT3, NF-κB, NRF2-AMPK, PI3K/AKT, and SIRT1, to regulate inflammation and cell death. The balance between oxidative and antioxidative processes is critical for therapeutic efficacy. Notably, RSV-induced ROS-mediated cell death, particularly in the context of TP53 mutations, represents a promising target for future interventions. Overall, RSV demonstrates multi-target potential for cancer and neurodegenerative disease therapy, though optimization of its pharmacological profile remains essential.
The milk thistle plant (Silybum marianum) is known for its hepatoprotective properties. However, the poor water solubility of silymarin limits its dissolution in the intestinal tract and restricts its bioavailability following oral administration. To improve bioavailability, special formulations, in particular micellar solubilization, are explored. In this study, we examined the transport rate of silymarin in a krill oil-based formulation across a Caco-2 epithelial barrier after upstream digestion simulation in vitro. Furthermore, in a randomized cross-over design study the bioavailability of the krill oil-based formulation was investigated after single dose intake in fasting conditions in healthy participants. We could demonstrate that the apparent transport coefficient of silybin, measured as lead substance of milk thistle extract, across the epithelium is efficiently boosted by a krill oil formulation, resulting in a 28% increase compared to silymarin powder. Consistent with these findings, a significant enhancement of bioavailability (P < 0.0001) was demonstrated for the krill oil-based formulation in comparison to the milk thistle extract resulting in an 8.59-fold higher AUC0-8h and a 15.08-fold greater Cmax of silybin and faster uptake kinetic after single dose intake. These findings suggest that phospholipid-based delivery systems offer a promising strategy for improving the efficacy of lipophilic bioactives. Furthermore, the combination of krill oil with milk thistle extracts efficiently provides silybin, PUFAs, and choline, which are important nutrients contributing to liver and heart health.
Metabolic 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.
Dengue virus (DENV) transmission has greatly increased in the last decade, partly due to the geographical expansion of Aedes spp. mosquitoes. These arthropods are now found even in temperate climates, including Europe, where outbreaks have occurred. A better understanding of the life cycle is essential, as it may enable the development of specific treatments or therapies, which are currently lacking. Recent breakthroughs concerning the viral capsid (C) protein structure and function are encouraging. It is now clear that the C protein binds both to host lipid droplets and to the viral genome-interactions crucial for viral encapsidation and replication. Here, we investigate how DENV C associates with the viral genome. Using DENV C and single-stranded DNA sequences analogous to relevant genomic regions, we biophysically characterize their interaction. A decrease in fluorescence intensity and lifetime, as well as changes in the protein secondary structure, were observed upon interacting with single-stranded DNA. These results are consistent with molecular condensation, supporting a possible liquid-liquid phase separation contributing to DENV C-nucleic acid complex formation.
Accumulating evidence suggests that SUMOylation plays a crucial role in the progression and resistance of secondary hyperparathyroidism (SHPT). However, the precise mechanism of SUMOylation in SHPT remains unclear. We identified the potential role of SUMOylation in SHPT based on RNA sequencing data obtained from Gene Expression Omnibus (GEO) datasets. Clinical samples were used to verify the expression of SENP1, SUMO1, SUMO2, GRHL2, and vitamin D receptor (VDR) in SHPT cells and tissues. Primary cells were extracted for subsequent experiments. Plasmid transfection and small interfering RNA (siRNA) were used to modulate SENP1 expression in SHPT primary cells. VDR relative expression was detected by Western blot (WB) and immunofluorescence. The effects of SENP1 on SHPT cell apoptosis and anti-proliferation were analyzed by flow cytometry and WB. Co-immunoprecipitation (Co-IP), chromatin immunoprecipitation (ChIP), and ChIP-quantitative polymerase chain reaction (ChIP-qPCR) were employed to explore the regulatory mechanisms of SENP1 in SHPT. We found that SUMOylation was significantly upregulated in SHPT and was closely related to calcitriol resistance. SENP1 and GRHL2 were downregulated. SENP1 was found to upregulate VDR and downregulate the SUMOylation of VDR, which mediates SENP1's regulation of SHPT cell apoptosis and anti-proliferation. Mechanistically Co-IP assays revealed binding between VDR, SENP1, SUMO1, and SUMO2. ChIP assays indicated that transcription of SENP1 was regulated by GRHL2, with binding sites identified by ChIP-qPCR. Additionally, we identified the potential binding pocket of SENP1 and screened 10 candidate small-molecule drugs approved by the US Food and Drug Administration (FDA). Our findings indicate a distinct mechanism of SENP1-mediated VDR SUMOylation and establish the critical role of the GRHL2/SENP1/VDR signaling axis in SHPT development.
Vestigial-like family member 3 (VGLL3), a transcriptional cofactor of the TEA domain family, has been previously identified as a regulator of osteoblast differentiation. Building upon our previous findings, we investigated VGLL3 function in MC3T3-E1 osteoblasts using an integrated approach combining transcriptomic analysis and functional assays to identify its downstream effectors and explore associated autophagy mechanisms. RNA-seq analysis of Vgll3-knockdown (shVgll3) cells identified death-associated protein kinase 2 (DAPK2), a regulator of autophagy, as a downstream effector. Autophagic activity was examined using transmission electron microscopy and western blot analysis of LC3-II and p62 proteins. The effects of Dapk2 knockdown (shDapk2) on osteoblast differentiation were evaluated using qPCR, western blotting, alkaline phosphatase staining, and Alizarin Red staining. Rapamycin treatment was used to determine whether pharmacologic activation of autophagy could restore osteoblast function. Vgll3 knockdown significantly suppressed autophagic flux, as evidenced by fewer autophagic vacuoles, decreased LC3-II accumulation, and increased p62 expression. A comparable reduction in autophagic activity was observed in shDapk2 cells and was accompanied by impaired osteoblast differentiation. Rapamycin treatment partially restored autophagy and osteogenic differentiation in Vgll3-deficient cells. Finally, overexpression of DAPK2 partially rescued autophagic activity and osteogenic differentiation in shVgll3 cells, supporting its role as a key downstream functional effector. FOXM1 was further implicated as a potential transcriptional regulator contributing to DAPK2 expression. Collectively, our findings suggest that VGLL3 may influence osteogenic differentiation in osteoblasts, potentially involving DAPK2-associated autophagy.
Electrical stimulation (ES) has emerged as a promising technique in the field of bioengineering and biomedicine, particularly in bone regeneration and cell differentiation. ES using alternating current (AC) is based on the periodic reversal of current direction, which generates oscillating electric fields. The application of an electric field has effects on cell growth and differentiation, as well as on morphology and migration. This study aimed to explore the effect of applying AC electrostimulation within the proliferation, differentiation, and morphology process of osteoblastic cells. The electrical stimulation signals were daily applied for 3 h during 14 days. Different frequencies were tested (1 Hz, 10 Hz, 100 Hz, and 1 kHz), with amplitudes of 125, 250, 500, 750, 1000, and 1500 mV/mm. Cell viability was estimated using the AlamarBlue, and MC3T3-E1 differentiation levels were evaluated through alkaline phosphatase (ALP) activity. RUNX2, OSX, ALP, OPG, and RANKL gene expression was assessed by RT-PCR. Morphological analysis was performed through cell transfection followed by immunofluorescence. Statistical analysis was conducted by SPSS.23 and graphs generated through Graph-pad. Viability and ALP activity were optimal at 10 Hz. Once the frequency was defined, RUNX2, OSX, ALP, OPG, and RANKL gene expression revealed an increase in the differentiation and osteogenic activity levels at 10 Hz and 500-750 mV/mm. As well as, morphological studies showed an increase in the area, pseudopodia length, and numbers at 500 mV 10 Hz conditions. The optimal ES condition to differentiate MC3T3-E1 cells is 10 Hz 500-750 mV/mm. Electrostimulation has emerged as a promising technique in the field of bioengineering and biomedicine, particularly in bone regeneration and cell early maturation.
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. In vivo, 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.
Growing the hyperthermophilic archaeon Pyrococcus horikoshii OT-3 in medium supplemented with d-allo-Ile instead of l-Ile markedly upregulates the activity of broad substrate specificity amino acid racemase (BAR). In P. horikoshii genome, the BAR gene (PH0138) forms a cluster with PH0137, which encodes a putative transporter protein, and PH0140 that encodes a feast/famine regulatory protein (FFRP), involved in the transcriptional regulation of metabolic pathway genes. Here, we performed gene expression, protein-DNA interaction, and crystallographic analyses to elucidate the expression mechanism of the BAR cluster. Gene expression analysis revealed that d-allo-Ile simultaneously induces PH0138 and PH0137. Electrophoretic mobility shift assays demonstrated that the PH0140 protein binds to the PH0137 promoter in the presence of l-Ile, but this interaction is disrupted by d-allo-Ile, identifying PH0140 as a d-amino acid-responsive regulatory protein (DARP). The crystal structures of DARP bound to l-Ile and d-allo-Ile were compared with those of another FFRP family member, P. horikoshii FL11. Our findings reveal a dimeric arrangement of l-Ile-bound DARP resembling the DNA-bound (open) form of FL11, whereas d-allo-Ile-bound DARP corresponds to the DNA-unbound (closed) form. These conformational changes result from subtle alterations in hydrogen bonding around the coregulators. Furthermore, cultivation in d-allo-Ile substitution medium only impacted the expression of PH0138 and PH0137, and not the other genes, suggesting that DARP regulates a more limited gene set than FL11. Cumulatively, these results reveal a distinct mechanism by which an FFRP homolog controls d-amino acid utilization.
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.
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 in vitro 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.
Researchers have developed hybrid bionic platforms for odor detection, inspired by natural chemoreceptive systems, advancing artificial olfactory systems that recognize specific volatile compounds. Odorant binding proteins (OBPs) are small carrier proteins found in the olfactory organs of mammals and insects. When coupled with electrical transducers, OBPs act as recognition elements, converting chemical signals into electrical outputs. This enables the development of biological electronic noses that are based on biomimetics and aim for sustainability. The objective of this review is to provide a comprehensive and updated overview of OBP-based biosensors, with a particular focus on insect OBPs as biorecognition elements, and to critically examine their applications, advantages, and technological potential across different fields. OBP-based biosensors show strong promise in medical diagnostics, environmental monitoring, food quality, insect pest control, and security. Insects demonstrate remarkable sensitivity to specific odors which makes them excellent models for designing bioinspired biosensors. Compared to conventional methods, OBP-based biosensors offer significant advantages in terms of portability, rapid response, and cost-effectiveness. OBPs are remarkably stable under different environmental conditions and can bind both volatile and aqueous-phase molecules, enhancing their functional versatility. Moreover, they can be produced through biotechnological processes using renewable resources, supporting eco-friendly innovation. These advantages make OBPs ideal candidates for next-generation biosensors in fields requiring real-time and on-site chemical detection.
The formation of macrophage-derived foam cells is a critical factor in the development of atherosclerotic lesions. However, the mechanisms through which the cuproptosis pathway contributes to macrophage foam cell formation remain unclear. This study aimed to investigate the role and mechanisms of the cuproptosis inducer Elesclomol in atherosclerosis. Macrophages were exposed to ox-LDL to establish a foam cell model. Subsequently, CCK8, oil red O staining, and Western blot were used to investigate foam cell formation and the cuproptosis pathway in macrophages. Furthermore, proteomics analysis was employed to explore the molecular mechanisms underlying the effects of Elesclomol. The results demonstrated that Elesclomol promoted foam cell formation by increasing lipid accumulation in ox-LDL-treated macrophages, which was attributed to the inhibition of cholesterol efflux mediated by ATP-binding cassette transporters A1 and G1 (ABCA1 and ABCG1). Additionally, Elesclomol enhanced the cuproptosis pathway in macrophages, leading to increased intracellular ROS accumulation and consequent cell death. Mechanistically, the cytotoxic effects of Elesclomol were mediated through activation of the mitogen-activated protein kinase (MAPK) pathway and upregulation of metallothionein 2A (MT2A). Collectively, these findings underscore the critical role of cuproptosis in the pathogenesis of atherosclerosis and suggest that targeting this pathway may represent a promising therapeutic strategy for the disease.