Search Results (1,332)

Background: Neuropathic pain remains difficult to treat because current analgesics often provide insufficient efficacy or dose-limiting adverse effects. Nav1.7 is genetically validated as a key regulator of human pain sensation, but the development of selective small-molecule Nav1.7 inhibitors has been limited by the high similarity among voltage-gated sodium channel subtypes. Methods: We generated monoclonal antibodies selectively targeting Nav1.7, humanized them for therapeutic development, and evaluated their binding, selectivity, functional channel inhibition, systemic analgesic efficacy, and effects on neuronal activity in a rat model of partial sciatic nerve ligation-induced neuropathic pain. Results: The humanized antibodies showed high-affinity and selective binding to Nav1.7 and functionally inhibited the channel in cellular assays. After systemic administration to neuropathic pain model rats, the lead antibody produced robust analgesia lasting at least 96 h. Electrophysiological analyses demonstrated reduced mechanically evoked and spontaneous neuronal activity, and immunohistochemistry showed decreased mechanical stimulus-induced phosphorylation of extracellular signal-regulated kinase in dorsal root ganglion neurons. The antibodies did not impair physiological nociception or motor function under the tested conditions. Conclusions: These findings provide preclinical proof of concept that humanized anti-Nav1.7 antibodies can act as systemically administered, long-acting biologic analgesics for neuropathic pain while preserving normal nociceptive and motor functions. The clinical advancement of S-151128 further supports the translational potential of this modality. Full article

Almonds are widely cultivated in Mediterranean regions, and their processing generates by-products such as almond skins, which are often discarded or used in low-value applications, leading to economic and environmental concerns. These skins are rich in bioactive compounds like polyphenols and flavonoids, with putative protective effects against toxins. Fungi such as Aspergillus and Penicillium species can contaminate nuts and their by-products and produce neurotoxic metabolites, like cyclopiazonic acid (CPA). This study aimed to characterize the phenolic compounds in aqueous extracts of almond skin and evaluate their cytoprotective effects on the viability of human neuroblastoma cells (SH-SY5Y) under individual CPA exposure and simultaneous co-exposure with almond skin extract. The extracts were optimized for extraction conditions, and UPLC-Triple-TOF-MS/MS analysis identified epicatechin, quercetin and kaempferol as the predominant phenolic compounds. Also, cell viability results showed that CPA induced cytotoxic effects on SH-SY5Y cells in a concentration-dependent manner. However, cells exposed to almond skin extract, at various dilutions (from 1/4 to 1/16), significantly increased cell viability from 43% to 57% relative to the control. Moreover, when SH-SY5Y cells were simultaneously co-exposed to CPA (400–600 nmol/L) and almond skin extract (1/4 dilution), a partial attenuation of CPA-induced toxicity (from 9% at 400 nmol/L to 20% at 600 nmol/L) was observed when compared with CPA alone. These findings suggest cytoprotective potential of almond skin extract in an in vitro neuronal-like model which may be associated with their content of phenolic compounds, providing new insights into their action against the emerging mycotoxin CPA, which remains underexplored in food safety research. Full article

Accurately quantifying the expression of individual transcript isoforms remains a formidable challenge, especially in contexts such as neurodegenerative diseases and cancers, which are characterized by high isoform diversity. The present study introduces a junction-based method, named differential splicing frequency analysis (DSFA), which enables more sensitive detection of differential splicing using RNA-seq data. Unlike the existing exon-, isoform-, and event-based methods, DSFA quantifies splice junction usage. We applied DSFA to Alzheimer’s disease (AD)-associated genes through large-scale RNA-seq data mining. The present study is the first to establish that the APP770-, APP751-, APP695-, and APP752-encoding isoforms represent major isoforms of the APP gene. Three important findings are: (1) the APP752-encoding isoform exhibits immune cell specificity; (2) the relative proportion of the APP752-encoding isoform increases during the differentiation of induced pluripotent stem cells (iPSCs) into microglia, akin to the increase in relative proportion of the APP695-encoding isoform during iPSC differentiation into neurons; and (3) the APP751-encoding isoform predominates in both cancer and immune cells. Additionally, we identified APP/58417N and App/52804N as differentially expressed splice junctions in humans and mice, respectively. Through over-expression of U1 snRNA in human embryonic stem cell (hESC)-derived neurons, we found that U1 snRNA over-expression decreases the usage of APP/58417N in neurons, similar to the effects observed in AD samples. Our research highlights that the major isoforms of a gene can differ markedly in their expression across tissue and cell types. Full article

Background: Early endolysosomal and autophagic defects are among the earliest cellular alterations observed in Alzheimer’s disease (AD). However, the molecular mechanisms linking amyloid precursor protein (APP) metabolism to vesicle trafficking dysfunction remain incompletely understood. The APP-derived fragment C99 has emerged as a potential upstream mediator of intracellular toxicity, but its impact on organelle homeostasis and its modulation by metabolic interventions remain unclear. Methods: To investigate these mechanisms, we expressed human C99 in Drosophila neurons and examined intracellular pathology using ultrastructural analysis, fluorescent reporters of autophagy and mitochondrial turnover, and proteomic interactome mapping. The effects of the ketone body β-hydroxybutyrate (BHB) were evaluated to assess the impact of metabolic intervention. Results: Neuronal C99 expression induced pronounced vesicular abnormalities, impaired autophagic turnover, and disrupted mitochondrial quality control. Transmission electron microscopy revealed extensive accumulation of enlarged vesicular compartments, accompanied by reduced mitochondrial turnover and accumulation of aged mitochondria. BHB treatment restored autophagic cargo clearance, improved mitochondrial turnover, and normalized vesicular ultrastructure. These protective effects required neuronal ketone transport, indicating a neuron-intrinsic metabolic mechanism. Proteomic analysis of the C99-associated interactome revealed that ketone treatment remodels networks enriched for vesicle trafficking and proteostasis pathways. Network prioritization identified the retromer component VPS35 as a candidate regulatory hub. Functional analyses demonstrated that depletion of VPS35 abolished the BHB-dependent restoration of autophagy, mitochondrial turnover, and vesicle morphology. Conclusions: Ketone treatment restores mitochondrial quality control and autophagic homeostasis through a VPS35-dependent mechanism in C99-induced neurodegeneration. These findings provide mechanistic insight into how metabolic interventions may restore intracellular homeostasis in Alzheimer’s disease. Full article

Background: Progesterone (P4) is used as an antiseizure medication (ASM) to treat catamenial epilepsy, refractory to first-line drugs. P4 and other neurosteroids (NSs) are important regulators of multiple nervous system functions, including neuronal excitability and synaptic plasticity. In addition to their antiseizure properties, P4 and other NSs are also anti-inflammatory agents. Neuroinflammation is an important pathophysiological mechanism of epilepsy refractory to ASMs. Accordingly, we evaluated the ability of P4 to modulate neuroinflammation, using human microglia activated by lipopolysaccharide (LPS). Methods: Human microglia (HMC3) were stimulated for 3 h with LPS in the absence or presence of various concentrations of P4. Thereafter, levels of (i) toll-like receptor 4 (TLR4), (ii) the NLRP3 inflammasome, and (iii) pro-inflammatory cytokines were quantitated by real-time PCR and Western blot analyses. Phagocytic activity was also assessed using a phagocytosis assay employing fluorescent beads. Results: P4 treatment significantly reduced the microglial inflammatory state induced by LPS, which was mediated by upregulation of the TLR4- and NLRP3-axes. The protective effects of P4 were mediated by inhibition of Nuclear Factor kappa-light-chain-enhancer of activated B cells (NFκB) phosphorylation and reduced activation of Mitogen-Activated Protein Kinases (MAPK). The effects of P4 included a significant reduction in mRNA levels of the main pro-inflammatory cytokines and a reduction in phagocytic activity of HMC3. Conclusions: P4 is endowed with significant anti-inflammatory properties, which may be involved in the beneficial effects reported for drug-resistant catamenial epilepsy. Further research is required to clarify P4 post-receptor mechanisms of action and to explore the roles of other P4-derived NSs. Full article

α-synuclein (α-syn) aggregation in dopaminergic neurons is a central event in Parkinson’s disease (PD) pathogenesis. Immunotherapeutic strategies targeting α-syn, including passive and active approaches, aim to inhibit aggregation, propagation, and toxicity of pathological species while promoting their clearance via immune mechanisms. This review summarizes α-syn directed immunotherapies evaluated in in silico, in vitro, and in vivo models, as well as early phase clinical trials, focusing on how epitope selection and antibody formats influence efficacy, safety, and target engagement. Data on monoclonal antibody, peptide, and protein-based vaccines, and structure-guided immunogens were analyzed, integrating behavioral, neuropathological, proteomic, and structural outcomes alongside biomarker development for α-syn species in cerebrospinal fluid and peripheral compartments. Clinical evidence indicates that several candidates induce sustained anti-α-syn antibody responses with acceptable safety profiles and signs of pharmacodynamic engagement, including reductions in free or oligomeric α-syn. However, consistent long-term clinical benefits remain unproven, highlighting the gap between preclinical success and disease modification in humans. Advances in structural biology and proteomics support rational epitope selection and improved immunogen design, reinforcing α-syn-targeted immunotherapy as a promising yet experimental strategy for PD, and highlighting the need for mechanistically oriented, biomarker-driven clinical trials initiated in well-characterized prodromal and early-stage cohorts. Full article

Human herpesvirus-6 consists of a pair of viral species, HHV-6A and HHV-6B, which are neurotropic with the ability to invade, persist, and reactivate within the nervous system. Accumulating evidence links HHV-6 to epilepsy and other neuropathologies, including: multiple sclerosis, chronic fatigue syndrome, and neurodegeneration. Yet, mechanisms by which these viruses induce neurological disorders, including their role in epileptogenesis, remain unknown. It has been demonstrated that HHV-6 exhibits tropism for astrocytes, oligodendrocytes, and neurons. Thus, HHV-6 can perturb cellular homeostasis, neuronal signaling, and immune regulation, astrocytic glutamate clearance, GABAergic inhibition, and cholinergic or monoaminergic neurotransmission yielding network hyperexcitability. It is also reported that HHV-6 can activate neuroinflammation through Toll-Like Receptor (TLR), cytokine, and/or NF-$\kappa$B activation, which facilitates neuronal injury and network instability. Indeed, a suite of converging processes suggest a multifactorial nature for HHV-6 related neuropathology. Despite robust experimental and clinical data, definitive causal relationships between HHV-6 and epilepsy (or induction of neurodegeneration) remain elusive. This review discusses evidence for roseolovirus-induced neurological dysfunction and disorders commonly associated with HHV-6A and HHV-6B infections. A preponderance of clinical and experimental evidence suggests that differential tropism for distinct neuronal neurotransmitter chemotypes and glia as well as systemic effects are involved in roseolovirus-mediated neurological disease. Full article

Alzheimer’s disease (AD) is a progressive neurodegenerative disorder characterized by progressive memory impairment and cognitive decline. The APOE4 allele represents one of the most prominent genetic risk factors. In this study, we investigated the impact of APOE4 on the cholinergic neuronal development and on the neuronal inflammatory response to TNF-α stimulation. To address this, human induced pluripotent stem cells (hiPSCs) carrying a homozygous APOE4 genotype and an isogenic APOE3 control were differentiated into cholinergic-like induced neurons (iNs) by LHX8 overexpression. APOE4 was associated with accelerated early neuronal differentiation, as reflected by earlier downregulation of the progenitor marker Nestin. However, delayed expression of synaptophysin indicated impaired synaptic maturation. Functionally, APOE3 iNs exhibited a robust but temporally regulated response to TNF-α, whereas APOE4 iNs were characterized by a delayed yet sustained induction of inflammatory signaling. Moreover, APOE4 iNs displayed an enhanced stress-associated transcriptional response at early differentiation stages. Collectively, these findings suggest that APOE4 influences both neuronal development and the timing and persistence of inflammatory responses, potentially predisposing cholinergic neurons to later dysfunction in AD. Full article

Extracellular vesicles (EVs) have emerged as key mediators of intercellular communication in the brain, with glial cell-derived EVs increasingly recognized for their roles in maintaining brain homeostasis and contributing to the progression of neurodegenerative diseases. By transferring a diverse cargo of bioactive molecules, including proteins, RNAs, and organelles, EVs influence recipient cell behavior and overall brain function. In neurodegenerative conditions, glial EVs can either propagate pathogenic signals or deliver neuroprotective and regenerative cues, depending on their cellular origin and molecular composition. This context-dependent heterogeneity highlights the need for physiologically relevant human models to investigate EVs biology. Human induced pluripotent stem cell (iPSC)-derived glial models provide a disease-relevant platform, as they recapitulate key pathological features of Alzheimer’s disease (AD), Parkinson’s disease (PD) and amyotrophic lateral sclerosis (ALS). When further integrated with brain organoid platforms, these iPSC-based systems enable the generation of three-dimensional environments that closely resemble in vivo EVs dynamics. Importantly, glial EVs can modulate cellular pathways involved in neuronal survival and function. Indeed, their potential to interact with and, under specific experimental conditions, traverse the blood–brain barrier (BBB) has contributed to growing interest in their application for biomarker discovery and therapeutic development. Engineered and patient-specific EVs derived from iPSCs are emerging as promising tools for targeted, cell type-specific, therapeutic approaches, although their clinical applicability still requires further validation. This review discusses the emerging evidence supporting the dual role of iPSC-derived glial EVs in health and disease, underscores the translational potential of iPSC-based platforms for mechanistic studies, and outlines their promise as precision medicine tools for diagnostics and therapy. Full article

The increasing prevalence of Alzheimer’s disease has created a substantial and urgent need for brain-healthy functional foods. The processing of Pacific white shrimp (Litopenaeus vannamei) generates considerable amounts of head waste, which is rich in bioactive compounds, including lipids and peptides, holding great promise for the development of nutraceuticals to support human brain health. However, traditional extraction methods are time-consuming and inefficient in fully utilizing these compounds. This study aimed to explore the functional properties of these shrimp head-derived ingredients using “one-step” three-phase partitioning (TPP) followed by successive proteolysis. The extracted polar lipid (PL-SH), protein (P-SH) and proteolytic peptidic product (Pep-SH) from shrimp heads were screened for their antioxidant, neuroprotective, and anti-neuroinflammatory activities. Antioxidant activities were evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS⁺), and hydroxyl free radical scavenging assays, all of which revealed strong antioxidant potential for all three products. Neuroprotective activities were assessed using HT-22 mouse hippocampal neuronal cells challenged with Aβ₂₅₋₃₅, and anti-neuroinflammatory activities were evaluated using BV-2 microglial cells stimulated with lipopolysaccharide (LPS). The results suggested that both PL-SH and Pep-SH exerted protective effects against Aβ₂₅₋₃₅-induced cell damage under the tested conditions, and PL-SH also reduced nitric oxide (NO) production induced by LPS, indicating potential anti-neuroinflammatory activity. However, further studies with additional biomarkers (e.g., ROS, apoptosis markers, and cytokines) are required to confirm these effects. The lipid composition of PL-SH was further characterized by thin-layer chromatography and LC-MS/MS-based lipidomics, revealing various classes of phospholipids. Furthermore, analysis of the molecular weight distribution and sequences of peptides in Pep-SH revealed peptide sizes ranging from 70 to 1700 Da and a high degree of homology to known antioxidant and neuroprotective peptide sequences. These findings suggest that lipids and peptides from Pacific white shrimp heads possess valuable functional properties, supporting their potential use in the development of functional foods for neuroprotection and anti-neuroinflammation. Full article

Cyclopiazonic acid (CPA), a neurotoxin produced by Penicillium and Aspergillus genera, induces oxidative stress and neuronal damage, mechanisms implicated in neurodegenerative diseases. This study investigates the oxidative stress induced by CPA in SH-SY5Y human neuroblastoma cells, focusing on mitochondrial membrane potential, mitochondrial superoxide levels, ROS production, lipid peroxidation and gene expression. Additionally, the cytoprotective effects of extra virgin olive oil (EVOO) extract, along with its major polyphenols oleuropein (OLE) and tyrosol (TYR), were evaluated. CPA exposure increased mitochondrial superoxide levels and lipid peroxidation, reducing mitochondrial membrane potential, although no intracellular ROS generation was observed. Gene expression analysis revealed downregulation of antioxidant defense genes (nrf2, nos2, ho1, cat, keap1, nqo1, gpx1 and gsr), with the strongest repression observed for nos2 (93%), nqo1 (83%) and ho1 (79%) at the highest CPA concentration, consistent with oxidative stress markers. EVOO extract demonstrated protective effects, enhancing cell viability across all CPA assayed concentrations (400–600 nM). Conversely, TYR and OLE exhibited variable and concentration-dependent effects, also showing protection to a lesser extent, while EVOO extract proved to be more effective due to synergistic interactions among its phenolic components. Overall, CPA induces mitochondrial oxidative damage as a key mechanism of neurotoxicity, while EVOO phenolics mitigate this toxicity. Full article

Background/Objectives: Magnesium (Mg) is essential for neuronal maturation, yet its role in human cortical development remains poorly defined. Here, we investigated the effects of physiological (1 mM) and elevated (5 mM) concentrations of MgSO₄ and magnesium pidolate (MgPid) on human brain organoids co-cultured with an in vitro blood–brain barrier (BBB) model. Methods: Human brain organoids derived from induced pluripotent stem cells were co-cultured with an in vitro BBB system and treated for 4 days with either MgSO₄ or MgPid at physiological and elevated concentrations. Cortical organization was assessed by transmission electron microscopy and immunofluorescence analysis. Western blotting for neurotransmitter receptors and Mg transporters, quantification of intraorganoid Mg²⁺ levels, ELISA-based measurement of GABA and dopamine, and analysis of glutamate were performed. Results: High Mg exposure enhanced cortical stratification and neuronal organization, as shown by the localization of CTIP2 in the outermost layer and TBR2 in the inner layer, together with ultrastructural features consistent with advanced differentiation. Elevated Mg increased intraorganoid Mg²⁺ levels without altering Mg transporter abundance and selectively modulated neurotransmitter receptor expression: NMDA-R levels were reduced by MgPid, whereas GABA_A-R and GABA_B-R were upregulated, particularly in response to MgPid. Levels of glutamate, GABA, and dopamine remained unchanged. Conclusions: These findings identify Mg, especially in the form of MgPid, as a modulator of cortical architecture and inhibitory–excitatory receptor balance in human organoids, supporting its potential relevance for neurodevelopmental regulation and Mg-based therapeutic strategies. These results also support organoids as human-relevant, animal-free tools for neuroscience and neuropharmacological research. Full article

Mitochondrial dysfunction is a central feature of aging, driving bioenergetic decline, increased oxidative stress, and increased vulnerability to neurodegenerative diseases. Human induced pluripotent stem cells (iPSCs) and iPSC-derived neurons provide powerful models to study these processes. Ginkgo biloba extract GBE LI1370 (GBE) has demonstrated antioxidant and mitochondria-protective properties in preclinical models, including improvements in mitochondrial membrane potential, reduction in reactive oxygen species, and enhanced neuronal survival. However, its effects on mitochondrial function in human iPSCs and their differentiated derivatives in the context of aging have not yet been investigated. This study evaluated the mitochondrial protective effects of GBE (100 µg/mL) in an established iPSC-based model of aging and in neurons and astrocytes derived from aged iPSCs. Mitochondrial parameters, including ATP production, mitochondrial membrane potential (MMP), mitochondrial reactive oxygen species (mtROS), superoxide levels, and mitochondrial respiration, were assessed. Aged iPSCs exhibited reduced ATP production and MMP, together with increased mtROS and superoxide levels compared to young controls. Astrocytes derived from aged iPSCs also displayed mitochondrial dysfunction. Treatment with GBE for 24 h increased ATP production and MMP, reduced oxidative stress, and improved mitochondrial respiration in both young and aged iPSCs, as well as in aged iPSC-derived neurons and astrocytes. These preliminary donor-based findings support further investigation of GBE-associated mitochondrial responses in human donor-derived cellular models of aging and warrant validation in larger donor cohorts. Full article

Background/Objectives: Statins are currently one of the most commonly used cholesterol-lowering drugs. In recent years, in addition to their well-known effects on the cardiovascular system, statins have been shown to exert beneficial effects in the progression of various neuropsychiatric and neurodegenerative diseases. Methods: In this study, the effects of lovastatin on the function of α7-nicotinic acetylcholine (nACh) receptors expressed in rat hippocampus and Xenopus oocytes were investigated. Results: In whole-cell patch clamp studies in hippocampal neurons, 21-day chronic (20 mg/kg), but not acute (20 min), lovastatin treatment caused significant potentiation of choline (a selective agonist for α7 nACh receptors)-induced currents and choline-induced increases in GABA_A receptor-mediated currents. Further studies in Xenopus oocytes expressing human α7-nACh receptors indicated that 72 h pretreatment with lovastatin caused a significant increase in α7-nACh receptor function with an EC₅₀ value of 296 nM. Other statins, such as simvastatin and pravastatin, also potentiated α7-nACh receptors. Potentiation by lovastatin treatment was associated with a significant decrease in oocyte cholesterol content and was diminished by Go6983, an inhibitor of protein kinase C (PKC), suggesting that both decreased cholesterol levels and activation of PKC are involved in statin potentiation of α7-nACh receptors. Conclusions: In conclusion, our findings indicate that chronic lovastatin treatment potentiates the function of α7-nACh receptors in hippocampal neurons and in Xenopus oocytes expressing human α7-nACh receptors and provides important insights that could guide future efforts to design novel drugs targeting α7-nACh receptors. Full article

Background/Objectives: Oxidative stress is a major contributor to cellular injury in many pathological conditions, including neurodegenerative disorders. Resveratrol, a natural polyphenol with antioxidant properties, has been proposed as a cytoprotective compound, although the molecular mechanisms underlying its effects remain incompletely understood. Here, we investigated whether the protective action of resveratrol against hydrogen peroxide-induced oxidative stress is mediated by adenosine receptor signalling and activation of the Nrf2 pathway in HeLa cells. Methods: Cells were treated with resveratrol alone or in combination with selective adenosine receptor antagonists and oxidant challenge, and cell viability, ROS production, receptor involvement, and Nrf2 expression and localization were analyzed. Results: Resveratrol at a non-toxic concentration significantly protected HeLa cells against oxidative damage, reduced ROS accumulation, promoted Nrf2 nuclear translocation and gene expression, and enhanced the gene expression of antioxidant enzymes such as SOD1, catalase, HO-1, and NQO1. Pharmacological blockade of the A_2A receptor prevented this protective effect, whereas the inhibition of A₁ and A₃ receptors enhanced it and avoided the increased SOD1, catalase, HO-1, and NQO1 gene expression promoted by resveratrol alone. Moreover, A_2A antagonism was associated with reduced PKA levels, consistent with the involvement of the cAMP/PKA signalling axis. Conclusions: Taken together, these observations support a model in which adenosine A_2A receptor signalling contributes to resveratrol-associated cytoprotection and Nrf2 activation in a human non-neuronal cell model. Our findings therefore provide mechanistic insight into resveratrol–adenosine receptor interactions and generate hypotheses to be tested in disease-relevant neuronal systems. Full article