In this narrative review we summarise some of the roles played by glutamate and its receptors in both epilepsy and schizophrenia, before presenting the kynurenine pathway of tryptophan metabolism as a generator of compounds acting at least partly by modulating those receptors. Increasing evidence indicates that epilepsy involves interactions between neuronal excitability, cellular metabolism and immune system signalling. The kynurenine pathway of tryptophan metabolism occupies a central position within this neuroimmune–metabolic network through its regulation of glutamatergic neurotransmission and both innate and adaptive arms of the immune response. The modulation of glutamate receptors is achieved by activating NMDA-sensitive sites by quinolinic acid and the blockade of all glutamate ionotropic receptors by kynurenic acid. In addition, emerging evidence is discussed which implicates the GRIA3-mediated AMPA receptor subunit GluA3 function in epilepsy and schizophrenia, and observations suggesting antagonistic actions of 3-hydroxykynurenine at kainate receptors. The evidence reviewed suggests that the modulation of the kynurenine pathway metabolism may be relevant to the development of agents which increase understanding of the mechanisms involved in epilepsy, while also addressing the psychiatric and systemic inflammatory disorders that frequently accompany it. A dysregulation of neuroimmune metabolism represents a framework for improved understanding of epilepsy and its associated neuropsychiatric comorbidities, while highlighting opportunities and challenges for therapies targeting glutamatergic and immunometabolic pathways.
Cervical cancer remains a major cause of cancer-related mortality worldwide, highlighting the need for strategies that may improve responses to conventional chemotherapeutics. Epigallocatechin gallate (EGCG), a green-tea polyphenol with diverse biological activities, has been investigated as a potential chemosensitizing agent. This study evaluated the interaction between EGCG and doxorubicin (DOX) in HeLa cervical cancer cells, with HaCaT keratinocytes included as a non-malignant comparator. Cell viability and drug interactions were assessed using the CCK-8 assay and Chou–Talalay combination index (CI) analysis. Complementary assays evaluated membrane integrity, wound closure, apoptosis, cell-cycle distribution, intracellular DCF-associated fluorescence with or without N-acetylcysteine (NAC) pretreatment, caspase-3 immunoreactivity, and EGFR, FOXP3, CASP3, and CASP7 mRNA expression. Network-based analyses were additionally used to identify candidate molecular associations and pathways. CI analysis demonstrated synergistic EGCG–DOX interactions in HeLa cells under the tested conditions, whereas additive or antagonistic interactions predominated in HaCaT cells. Combined treatment produced the greatest reduction in viable cells, increased apoptosis, altered cell-cycle distribution, and reduced wound closure. It also produced the highest viability-normalized DCF-associated fluorescence, which was attenuated by NAC pretreatment, indicating an antioxidant-sensitive change in intracellular oxidative status without establishing a causal role in cytotoxicity. Combined treatment was further associated with increased total caspase-3 immunoreactivity and altered EGFR, FOXP3, CASP3, and CASP7 transcript levels. Overall, EGCG and DOX exhibited synergistic interactions and multiple treatment-associated cellular and transcriptional responses in HeLa cells under the present in vitro conditions. These findings do not establish cancer-specific selectivity or a definitive molecular mechanism but provide a basis for validation in additional cervical cancer models and at clinically relevant exposures.
Trihelix is a class of transcription factors unique to plants that play a major role in abiotic and biotic stress responses, seed isolate development, floral organ morphogenesis, and plant photomorphogenesis. Nevertheless, the Trihelix transcription factor family in Gardenia jasminoides (G. jasminoides) has not been systematically characterized. In this study, 11 GjTrihelix genes were identified from the G. jasminoides genome, unevenly distributed across five chromosomes, and can be classified into four subfamilies: GT-1, GT-2, SIP and SH4. Gene structure and functional motif analyses revealed high conservation within the same subfamily. Cis-acting element analysis showed that these genes are closely related to hormone responses, stress responses, and growth and development processes. Intraspecific synteny analysis showed a segmental duplication between GjTrihelix-3 and GjTrihelix-10. Interspecific collinearity analysis revealed that G. jasminoides shared 21 collinear gene pairs with soybean, compared with five pairs with Arabidopsis and 13 with Populus, indicating greater syntenic block conservation between G. jasminoides and soybean. Transcriptome data analysis demonstrated distinct spatiotemporal expression specificity of this gene family. Several genes were constitutively expressed in fruits; GjTrihelix-1 and GjTrihelix-3 were predominantly expressed in green fruits, while GjTrihelix-11 was highly expressed in red fruits. Under melatonin treatment, five GjTrihelix genes showed significant up-regulation and obvious transcriptional suppression of another five genes. Following infection by Botryosphaeria dothidea, GjTrihelix-5 and GjTrihelix-7 were progressively induced and peaked at 72 h. qRT-PCR results indicated that most GjTrihelix genes were highly expressed in leaves, while GjTrihelix-11 was highly expressed in flowers. Most GjTrihelix genes were significantly down-regulated under NaCl, ABA, GA3 and IAA stresses. This study provides new insights into the potential association of the Trihelix transcription factor family in G. jasminoides growth, development, and stress adaptation, offering theoretical references for stress-resistant G. jasminoides breeding.
Bufalin is a naturally occurring bufadienolide with broad-spectrum anticancer activity. Unlike conventional cytotoxic agents, bufalin exerts pleiotropic antitumour effects by directly modulating oncogenic proteins and promoting their degradation. It also disrupts metabolic plasticity, induces multiple forms of regulated cell death, counteracts therapeutic resistance, and remodels the immunosuppressive tumour microenvironment. However, the further application of bufalin is hindered by poor aqueous solubility, rapid systemic clearance, a narrow therapeutic window, and dose-limiting toxicity. Nanodrug delivery systems (NDDSs) offer a promising strategy for translating these interconnected pharmacological effects into spatially and temporally controlled therapeutic responses. This review summarises the distinctive anticancer mechanisms of bufalin and systematically evaluates the organic, inorganic, biomimetic, and hybrid nanocarriers developed for its delivery. Particular attention is given to tumour-responsive drug release, the targeting of cancer stem cells, the induction of ferroptosis and pyroptosis, metabolic modulation, sensitisation to phototherapy, and the activation of antitumour immunity. Finally, the major translational challenges are critically examined to inform the further rational development of bufalin-based nanomedicines.
Uterine corpus endometrial carcinoma (UCEC) is the most common gynecologic malignancy, and aberrant canonical NF-κB signaling is implicated in uterine corpus endometrial carcinoma (UCEC), yet the expression, epigenetic, and prognostic profile of IκB kinase (IKK) complex subunits CHUK (IKKα), IKBKB (IKKβ), and IKBKG (NEMO) remains undefined. This study assessed expression, methylation, miRNA regulation, and clinical relevance of IKK-complex genes in UCEC using public datasets. RNA-seq expression (TCGA-UCEC; GEPIA2) showed significant downregulation of IKBKB in tumors versus normal endometrium (unpaired Wilcoxon), whereas CHUK and IKBKG showed non-significant increases; immunohistochemistry (Human Protein Atlas) illustrated heterogeneity and suggested possible mRNA–protein discordance for IKBKB. Promoter methylation analysis (UALCAN; Illumina 450K) identified CHUK hypomethylation and IKBKG hypermethylation in tumors, with no significant change for IKBKB. Stratification showed IKBKB suppression across clinicopathological strata and TP53 mutant/wild-type tumors; IKBKG decreased across subtypes and stages. Pan-cancer profiling (TIMER2.0) highlighted IKBKB as the broadly dysregulated IKK member across malignancies. STRING networks indicated connectivity with NF-κB mediators (e.g., RELA, NFKB1, TRAF6). miRNA predictions (miRDB/TargetScan) revealed shared regulation (CHUK–IKBKB: 14 miRNAs; CHUK–IKBKG: 2; IKBKB–IKBKG: 0). Kaplan–Meier analysis indicated that elevated IKBKG expression correlated with reduced overall survival (HR = 1.85, p = 0.0037), while CHUK expression did not correlate with survival, and IKBKB exhibited a non-significant trend. In a multivariable Cox regression analysis, high IKBKG expression continued to show a significant association with reduced overall survival, even after adjusting for age, histological type, histological grade, and TCGA molecular subtype (adjusted HR = 1.58, 95% CI = 1.00–2.47, p = 0.048). These findings collectively suggest the potential prognostic significance of IKBKG in UCEC, although independent validation in larger, comprehensively annotated cohorts is still required. Collectively, these findings suggest IKBKG as a candidate negative prognostic marker and indicator in UCEC, warranting experimental validation.
Allergic asthma is characterized by Th2-driven airway inflammation. Indoleamine 2,3-dioxygenase (IDO) maintains immune tolerance, but its role in modulating the anti-inflammatory actions of Curcumin (CN) is unclear. This study investigated whether CN retains efficacy in a rat model of Ovalbumin (OVA)-induced airway inflammation under pharmacological IDO inhibition. Rats were sensitized and challenged with OVA and treated with methylprednisolone (MP, 15 mg/kg), CN (200 mg/kg), the IDO inhibitor 1-MT (70 mg/kg), or CN+1-MT. Assessments included systemic and pulmonary leukocyte profile, delayed-type hypersensitivity (DTH), OVA-specific IgE, bronchoalveolar lavage fluid (BALF) inflammatory cells and nitric oxide (NO) levels, lung wet/dry weight ratio, gene expression of tumor necrosis factor alpha (TNF-α), interleukin (IL)-4, IL-6, Cxcr2 and transcription factor Nfkb1 and lung histopathology. CN alone or combined with 1-MT normalized leukocyte counts, suppressed eosinophilic and neutrophilic infiltration, restored BALF NO levels, reduced DTH response and OVA-specific IgE, decreased pulmonary edema, and preserved pulmonary vascular integrity. CN significantly attenuated TNF-α, IL-4, IL-6, and Cxcr2 gene expression despite pharmacological IDO inhibition. Histopathology revealed reduced inflammatory infiltration and maintained alveolar structure. CN maintains anti-inflammatory, immunomodulatory and tissue-protective efficacy in allergic airway inflammation despite pharmacological IDO inhibition. These findings call for future research on CN as a potential adjunctive therapy method for inflammatory airway diseases. However, to elucidate the underlying processes, more research on IDO activity and downstream immunometabolic pathways is needed.
Microbial lipids produced by oleaginous yeasts are a sustainable alternative to conventional lipid sources because of their ability to generate nutritionally valuable fatty acids. This study evaluated the effects of carbon-to-nitrogen (C/N) ratio, modified through molasses concentration, and cultivation temperature (15 and 25 °C) on biomass production, lipid accumulation, fatty acid composition, and nutritional quality of lipids produced by the native yeast Rhodotorula mucilaginosa 6S. Biomass, lipids, sugar consumption, and growth kinetics were determined, while fatty acid composition was analyzed by gas chromatography and nutritional quality was assessed through lipid nutritional indices. Biomass production was mainly influenced by the C/N ratio, whereas temperature had the greatest effect on lipid accumulation and fatty acid composition. Cultivation at 25 °C enhanced lipid yield and productivity, while 15 °C promoted faster growth and higher accumulation of polyunsaturated fatty acids (PUFAs), resulting in lipids with superior nutritional quality. The combination of 15 °C and a C/N ratio of 12 produced the most favorable lipid profile, with higher unsaturation, improved PUFA/SFA and MUFA/SFA ratios, balanced ω-6/ω-3 and LA/ALA ratios, low atherogenicity and thrombogenicity indices, and a high hypocholesterolemic/hypercholesterolemic ratio. These findings demonstrate the potential of molasses as a sustainable substrate for producing high-quality microbial lipids.
The continual spermatogenesis throughout adulthood is ensured by a rare and unique cell group named spermatogonial stem cells (SSCs), which undergo self-renewal and/or differentiate into sperm cells. SSCs also become a promising genetic source for the protection of animal biodiversity. However, the isolation and culture of SSCs in vitro is still a big challenge and poorly explored in rabbits. The main objective of this study was to isolate, culture, and deeply characterize SSCs obtained from rabbit testes. Briefly, rabbit testicular tissue was mechanically and enzymatically dissociated, and obtained testicular somatic and germ cells were cultured for a short term in culture media supplemented with specific molecular factors maintaining SSC self-renewal and proliferation (GDNF, GFRα-1, FGF2, etc.). Immunofluorescent and PCR techniques were used for molecular profiling of cultured SSCs, while TEM analysis revealed their ultrastructure. After a few weeks, round and grape-like SSC colonies emerged, growing on the feeder cell layer. Rabbit SSCs showed positive staining for DBA, GFRA1, PLZF, RET, PGP9.5, DAZL, and DDX4. Increased expression of additional SSC markers was noticed using RT-qPCR and dd PCR (RET, PLZF, PGP9.5, DAZL, DDX4, CDH1, CD9, CD14, CD90, c-kit, ALDH, SSEA-4, SALL4, OCT4, and SOX2), while ultrastructure typical for primitive undifferentiated cells was observed under TEM. In conclusion, we successfully established a method for rabbit SSC isolation, culture, and phenotyping, which might facilitate their collection for further cryopreservation. However, the self-renewal, proliferative, and differentiation capacities of cultured SSCs still need to be confirmed through an in vivo SSC transplantation experiment.
Bisphosphonate-related osteonecrosis of the jaw (BRONJ) remains a challenging complication of bisphosphonate therapy because jaw extraction sockets exposed to bisphosphonates represent impaired wound environments rather than ordinary bone defects. This narrative review summarizes clinical, cellular, animal, and biomaterial evidence on the mechanisms that limit BRONJ repair and discusses how these pathological barriers can inform local material design. Current evidence suggests that BRONJ repair is constrained by impaired osteoclast-mediated remodeling, osteocyte and osteoblast dysfunction, oxidative stress, unresolved inflammation, angiogenic insufficiency, microbial challenge, mucosal instability, and changes in bone material properties. Biomaterial strategies investigated to date include local delivery of regenerative factors, restoration of remodeling activity, extracellular vesicles, nucleic acid nanostructures, platelet-derived matrices, antibacterial and ion-releasing hydrogels, angiogenic or lymphangiogenic systems, and mechanically adaptive scaffolds. Most studies remain preclinical and are based on rodent extraction or mandibular defect models, and few establish a direct causal link between a specific material property and durable BRONJ resolution. Future materials should be judged not only by their ability to enhance bone formation, but also by whether they can re-establish a sealed, vascularized, immune-balanced, and remodeling-competent socket capable of sustained jawbone repair.
Background: Fetal growth restriction (FGR) is associated with placental dysfunction and adverse perinatal outcomes. Although insulin-like growth factor-1 (IGF-1) signaling is important for placental and fetal development, the mRNA expression, immunopositivity and potential biological significance of specific IGF-1 isoforms in FGR remain incompletely characterized. This study investigated placental IGF-1Eb mRNA expression and immunopositivity in FGR pregnancies compared with appropriate-for-gestational-age (AGA) pregnancies. Methods: A total of 62 third-trimester human placentas were analyzed, including 47 from pregnancies complicated by FGR and 15 from pregnancies with AGA fetal growth. The mRNA expression of the IGF-1Eb isoform was assessed by reverse-transcription quantitative PCR in a subset of 28 fresh placental samples. IGF-1Eb protein immunoreactivity was assessed in paraffin-embedded tissue sections. Histopathological lesions were classified according to the Amsterdam criteria, and associations with clinical, demographic, and pathological parameters were evaluated using appropriate statistical analyses. Results: IGF-1Eb mRNA expression did not differ significantly between the FGR and AGA groups. In contrast, significant differences in IGF-1Eb immunoreactivity were observed in selected placental compartments. Moderate immunoreactivity in the perivillous syncytiotrophoblast was more frequent in FGR placentas and was associated with histological features of maternal vascular malperfusion, as well as with gestational age, neonatal birth weight, placental weight, maternal body mass index, and fetal sex. Increased IGF-1Eb immunopositivity was also observed in the endothelium of maternal decidual and fetal villous vessels in FGR placentas. No significant differences were observed in IGF-1Eb immunoreactivity in the extravillous trophoblast. Conclusions: In this observational study, placental IGF-1Eb protein immunoreactivity, but not mRNA expression, differed between FGR and AGA pregnancies in selected placental compartments. These findings indicate compartment-specific differences in IGF-1Eb protein immunoreactivity, associated with FGR and placental pathological features. However, the observational and cross-sectional design does not establish causality or a functional role for IGF-1Eb in placental dysfunction. Larger prospective studies incorporating functional validation are required to clarify the biological significance of these findings.
Chemotherapy-Induced Peripheral Neuropathy (CIPN) is a dose-limiting complication of paclitaxel (PTX) therapy, for which effective treatments are still lacking. This study evaluated the neuroprotective potential of Thymoquinone (TQ), a bioactive derivative of Nigella sativa, in mitigating chronic PTX-induced neurotoxicity without compromising antineoplastic activity. We first performed in vitro experiments using Sprague–Dawley rat embryonic (E15) Dorsal Root Ganglia (DRG) (Envigo Laboratory (Udine, Italy)) to assess neurotoxicity through neurite outgrowth evaluation. To investigate the molecular mechanisms underlying TQ’s putative neuroprotective mechanisms, SIRT1 protein expression was additionally evaluated by western blot, while MCF-7 and MDA-MB-231 breast cancer cells were used to monitor cytotoxicity via MTT assay. We then moved to in vivo experiments, in which chronic neuropathy was induced in rats using PTX (10 mg/kg, i.v., weekly for 4 weeks). TQ was co-administered orally (5–10 mg/kg/day). The effects of TQ on peripheral neuropathy were assessed through behavioral testing, neurophysiological assessments, and histological analysis of Intraepidermal Nerve Fibre density (IENF), DRG and peripheral nerves. In vitro, TQ (5 μM) significantly attenuated PTX-induced neurite shortening at 24 h; TQ co-treatment fully prevented PTX-induced SIRT1 downregulation in embryonic DRG neurons and did not compromise PTX cytotoxicity in MCF-7 cells, while significantly potentiating it in MDA-MB-231 triple-negative breast cancer cells. In vivo results demonstrated that PTX-treated animals exhibited mild erythroid myelosuppression at the end of treatment. Regarding efficacy, TQ consistently prevented PTX-induced mechanical allodynia throughout the treatment period, and TQ (10 mg/kg) transiently mitigated IENF depletion at mid-treatment; however, no improvement in neurophysiological parameters or peripheral nerve morphology was observed at either time point. Collectively, these findings suggest that, under conditions of chronic PTX exposure, TQ exerts a predominantly analgesic effect in vivo, without conferring meaningful structural neuroprotection against PTX-induced peripheral nerve degeneration.
Oxygen availability is a key regulator of organ maturation during the perinatal period. Disruption of physiological oxygen homeostasis contributes to prematurity-associated disorders, yet its effects on the coordinated maturation of the enteric nervous system (ENS) and gut microbiome remain poorly understood. Because β3-adrenergic receptor (β3-AR) signaling has emerged as a mediator of tissue adaptation to oxygen, we investigated whether activation of this pathway modulates hyperoxia-induced alterations in the developing colon. Newborn rats were exposed to normoxia or hyperoxia (85% O2) from birth to postnatal day 14 and treated with the β3-AR agonist BRL37344 (1 or 3 mg/kg). Enteric neuronal and glial populations were evaluated by quantitative immunofluorescence, whereas the colonic microbiome (CM) was characterized by 16S rRNA gene sequencing. Hyperoxia reduced neuronal density and altered neurochemical coding within the submucosal plexus, disrupted enteric glial organization in both the colonic submucosal plexus and mucosa, and remodeled the intestinal microbiome without affecting overall community diversity. BRL37344 treatment partially preserved submucosal neurochemical coding, modulated neuron–glia organization within the submucosal plexus, prevented the loss of mucosal enteric glial cells, and reshaped microbial composition. Collectively, these findings demonstrate that neonatal hyperoxia disrupts coordinated postnatal maturation of the ENS and CM and indicate that β3-AR signaling may contribute to postnatal intestinal adaptation to neonatal oxygen imbalance.
Schizophrenia is a highly heterogeneous neuropsychiatric disorder characterized by positive symptoms, negative symptoms, and cognitive impairment, with substantial long-term functional consequences. Although dopaminergic dysfunction remains central to current disease models and treatment, dopamine-centered frameworks alone cannot fully explain cognitive deficits, treatment resistance, or marked variability in therapeutic response. Emerging evidence supports a broader view in which genetic susceptibility and environmental exposures converge on multiple interacting biological systems, including dopaminergic and glutamatergic neurotransmission, neuroimmune and glial dysfunction, kynurenine pathway metabolism, large-scale brain network dysconnectivity, and gut–brain communication. In this review, we integrate these mechanisms within a systems-level framework and discuss how their interactions may contribute to symptom heterogeneity and disease progression. We further examine the limitations of conventional dopamine D2-based antipsychotics, emerging non-dopaminergic pharmacological strategies, and adjunctive interventions targeting cognition and functional recovery. Finally, we highlight major translational barriers, including treatment resistance, medication non-adherence, adverse-effect burden, and the lack of clinically actionable biomarkers. An integrated understanding of converging neurobiological mechanisms may provide a stronger foundation for mechanism-informed patient stratification and precision treatment in schizophrenia.
Acute lung injury (ALI) is a common clinical acute respiratory disorder driven primarily by a diffuse pulmonary inflammatory response. Cordycepin (COR) is a bioactive metabolite extracted from the fungus Cordyceps militaris, which possesses antioxidant and anti-inflammatory properties. However, its underlying molecular mechanism remains to be elucidated. This study evaluated the therapeutic effect of COR on ALI and the underlying molecular mechanisms. MH-S cells were primed with lipopolysaccharide (LPS) at 1 µg/mL for 24 h and then treated with varying doses of COR for an additional 24 h. WB and RT-qPCR analyses showed that COR inhibited the phosphorylation of TGF-β-activated kinase 1 (TAK1) as well as the key kinases in the MAPK and NF-κB pathways in LPS-induced MH-S cells, as evidenced by decreased TAK1, p38, JNK, IκB-α and P65 expression levels, as well as decreased TNF-α, IL-6, IL-1β, MAP3K7, MAPK8 and MAPK14 relative expression. Six-week-old BALB/c mice were intranasally instilled with LPS at 3 mg/kg, followed 24 h later by oral gavage administration of various concentrations of COR. The results showed that COR can effectively suppress the progression of pulmonary tissue injury; similarly, the expression levels of key proteins and inflammatory factors in the TAK1-MAPK and NF-κB signaling pathways were downregulated. In summary, COR exerts a therapeutic effect on ALI by directly targeting TAK1 to inhibit the activation of the MAPK and NF-κB signaling pathways and concurrently suppressing key inflammatory cytokines.
Cannabinoids are terpenophenolic compounds derived from Cannabis sativa L. that exert a broad range of biological and pharmacological activities. Increasing evidence highlights their potential as modulators of cancer progression specifically through the suppression of tumor cell growth, angiogenesis, and metastasis across multiple tumor models. This review provides a comprehensive overview of the molecular mechanisms by which natural and synthetic cannabinoids induce regulated cancer cell death. Current evidence demonstrates that cannabinoids regulate multiple forms of cancer cell death, including apoptosis, autophagy-dependent cell death, necroptosis, ferroptosis, and parthanatos. These effects are mediated through complex and interconnected signaling pathways such as TRIB3/AKT/mTORC1, PI3K/AKT/mTOR, MAPK/ERK, NF-κB, ERK/JNK/p38-MAPK, and ceramide/Raf1/ERK/ROS. In addition to their direct antitumor effects, cannabinoids can enhance the efficacy of conventional anticancer therapies through the coordinated regulation of complementary cell death pathways. They also provide clinically relevant supportive benefits in palliative care, alleviating chemotherapy-induced nausea, cachexia, and mood or sleep disturbances. Collectively, these findings identify cannabinoids as promising anticancer agents and therapeutic adjuvants, predominantly in the preclinical setting. However, significant challenges remain regarding their safety, optimal dosing, formulation, and clinical efficacy. Further mechanistic studies, rigorous preclinical research, and well-designed clinical trials are required to establish the translation of cannabinoid-based therapies into precision oncology.
The ubiquitous nicotinamide adenine dinucleotide (NAD) engages in diverse biological processes, leading to non-selective energy transfer toward target synthetic reactions. To achieve selective energy transfer in complex biological systems, we previously constructed artificial systems mediated by the non-natural cofactor nicotinamide cytosine dinucleotide (NCD), which can be specifically recognized by engineered enzymes with minimal cross-talk with natural cofactors. For enhanced energy transfer and higher product yields, efficient conversion of NCD to NCDH is required to deliver reducing power in NCD-mediated biosynthetic pathways. Here, we established a comprehensive strategy for selective reduction in intracellular NCD. First, coupled enzymatic colorimetric assays with high specificity were validated for quantifying NAD, NADP, and NCD. With phosphite as the energy source, we selectively elevated the intracellular NCDH/NCD ratio with minimal perturbation to NADH/NAD and NADPH/NADP ratios in E. coli. To overcome the limitation of phosphite transmembrane transport, cell-free systems were constructed to confirm that phosphite could drive near-complete NCD reduction. Finally, cells were treated with polymyxin B, which promoted phosphite uptake and thereby enabled maximal reduction in intracellular NCD. An NCDH/NCD ratio of 47 was achieved, demonstrating that 98% of the intracellular NCD pool existed in the reduced form. This work demonstrates that NCD can function as an independent redox cofactor for selective regulation, providing viable strategies for artificial cofactor-driven biosynthesis.
Spontaneous deamidation and isomerization of asparagine (Asn) residues is a major pathway of nonenzymatic protein aging, where Asn-Gly-containing peptides (NG motifs) represent the most reactive sequence. In this context, the first step of isomerization is succinimide formation, which is initiated by a concerted proton-transfer and nucleophilic activation step. Although the overall mechanism is established, the detailed nature of the rate-determining succinimide formation step and the possible role of nuclear quantum effects remain unclear. Here, we combine density functional theory (DFT), intrinsic reaction coordinate (IRC) analysis, and quantitative NMR kinetics to investigate hydrogen/deuterium (H/D) substitution effects on NG isomerization in different model peptides, which represent both conformationally flexible and restricted systems. Deuteration preserves the reaction pathway and structural evolution, indicating an invariant classical reaction coordinate. However, kinetic isotope effects in the flexible peptide system reveal a mixed classical–tunneling mechanism in the rate-determining step with observable deviations from classical over-the-barrier behavior. In contrast, the conformationally restricted peptide shows suppressed tunneling contributions. These findings demonstrate that conformational accessibility modulates proton tunneling in spontaneous peptide rearrangements and extends nuclear quantum effects beyond enzymatic systems to nonenzymatic processes associated with protein aging.
The neurovascular unit (NVU) is composed of a diverse array of cells and an extracellular matrix (ECM). Neural cells and blood vessels are intricately interconnected, forming a cohesive whole. Specific cellular components and structures within the NVU play an indispensable role in maintaining homeostasis of the central nervous system (CNS). With the advancement and maturation of cell co-culture technology, various three-dimensional (3D) NVU models continue to emerge, offering a more objective and comprehensive perspective for in vitro studies of CNS diseases. Specifically, these 3D NVU models include Transwell Chamber models, gel-polydimethylsiloxane (PDMS)-based 3D models, self-assembled NVU models and microfluidic NVU models, which reconstruct the complex NVU architecture to varying degrees. This review systematically summarizes multiple 3D construction strategies for in vitro NVU to overcome the limitations of conventional cellular tests or animal experiments, highlights the critical roles of biomimetic gel in recapitulating native cell-gel crosstalk, comparatively analyzes four major 3D NVU technical routes in terms of cellular composition, vascular morphology, barrier performance, and reproducibility, categorizes application scenarios of 3D NVU platforms oriented to practical research demands, including oxygen-glucose deprivation/reoxygenation (OGD/R) injury modeling, blood-brain barrier (BBB) permeability assay, CNS drug penetration screening, neuroinflammation and neurotoxicity evaluation, proposes practical principles for model selection under different experimental purposes, and concludes with current bottlenecks, including imperfect vascular network maturation and lack of unified evaluation criteria, together with future perspectives for standardized 3D NVU in vitro. By comparing the advantages and limitations of these approaches, we aim to clarify their optimal applicability for investigating specific pathological mechanisms and screening potential therapeutics.
Arterial hypertension is a key trigger for all major, life-threatening cardiovascular events such as arrhythmia, stroke, cardiac infarction and heart failure. Ethanol consumption has been identified as a critical risk factor, which promotes hypertension and thereby corrupts cardiovascular health and limits overall life expectancy. For heavy drinkers (≥72 g/day), reduction in half of the alcohol consumption reportedly results in considerable decreases in systolic and diastolic blood pressure levels amounting to 5.50 and 3.97 mmHg, respectively. Based on the complex network of ethanol effects on mammalian physiology, diverse mechanistic concepts have been postulated for a causal linkage between alcohol consumption and distorted blood pressure regulation. The impact of ethanol on the cardiovascular system involves a wide range of ethanol-sensitive cell types, ranging from neurons to vascular smooth muscle, endothelial and immune-cells, all of which express an interdependent array of potential ethanol target structures. Here, we provide an update on currently available evidence for a causal link between alcohol consumption and the etiology of arterial hypertension. The potential role of ethanol target structures with particular focus on membrane ion channels is discussed along with novel concepts of lipid-dependent modulation of signaling processes in the plasma membrane by ethanol.
Introduction: Thrombopoietin mimetics (TPOm) are known to increase blood cell counts, thereby ameliorating various diseases, including cardiomyopathy—a leading cause of mortality because of delayed radiotoxic effects. Previous preclinical studies demonstrated significant survival enhancement following lethal radiation exposure with a single dose of JNJ-26366821 in two mouse strains, CD2F1 and C57BL/6. Materials and Methods: In this study, a single dose of JNJ-26366821 (1.0 mg/kg) or saline was administered 24 h post-8.0 Gy total body irradiation (TBI) to male C57BL/6 mice. Mice were euthanized on days 1, 7, 15, and 30 post-TBI, and serum and heart samples were collected. Unirradiated mice treated with saline served as baseline controls. Inflammatory serum cytokines/chemokines, growth factors and multi-omics analyses, including proteomics, metabolomics, and lipidomics, were evaluated to assess therapeutic impacts. Results: JNJ-26366821 demonstrated overall positive effects, restraining radiation-induced inflammatory surges by 7 days post-TBI. Functional analysis revealed that JNJ-26366821 treatment in unirradiated mice activated cellular homeostasis, which persisted after post-exposure intervention in irradiated mice. The drug immediately reduced cell death, but networks supporting cardiovascular health, such as angiogenesis and vasculogenesis, remained inhibited 2 h post-intervention. By 7 days post-TBI, JNJ-26366821 mobilized molecules in heart tissue to reconstitute cellular development and maintenance. Protein synthesis and metabolism were activated between 15 and 30 days post-TBI, potentially compensating for radiation-induced muscle wasting. Conclusions: JNJ-26366821 demonstrated promising mitigating effects on cellular homeostasis and cardiovascular health when administered as a single dose at 24 h post-exposure.
