Diet-derived circulating antioxidants and risk of epilepsy: A study combining metabolomics and mendelian randomization.

Background: Previous studies offer inconclusive results on the association between diet-derived circulating antioxidants and epilepsy. Objective: This study aims to assess oxidative stress presence in epilepsy patients' circulation and investigate the causal link between diet-derived circulating antioxidants and epilepsy. Methods: Untargeted metabolomics analysis was conducted on plasma samples from 62 epileptic patients and 20 healthy individuals to evaluate oxidative stress based on metabolite alterations in epilepsy patients' circulation. Two-sample Mendelian Randomization (MR) analysis examined the causation between diet-derived circulating antioxidants (measured by absolute levels and relative metabolite concentrations) and epilepsy, utilizing the inverse-variance weighted (IVW) method as the primary outcome, with complementary MR analysis methods (MR Egger, weighted median, weighted mode, and simple mode). Results: Untargeted metabolomics analysis revealed elevated circulating oxidizing metabolites (palmitic acid, oleic acid, linoleic acid, and myristic acid) and reduced reducing metabolites (glutamine) in epilepsy patients, providing robust evidence of oxidative stress. The IVW analysis indicated significantly reduced epilepsy risk (odds ratio: 0.552; 95% confidence interval: 0.335-0.905, P = 0.018) with genetically determined higher absolute circulating ß-carotene. However, other diet-derived circulating antioxidants (lycopene, retinol, ascorbic acid, and selenium) and antioxidant metabolites (α-tocopherol, γ-tocopherol, ascorbic acid, and retinol) did not significantly associate with epilepsy risk. Additional MR analysis methods and heterogeneity assessments confirmed the results' robustness. Conclusion: This study provides compelling evidence of oxidative stress in epilepsy patients' circulation. However, the majority of diet-derived circulating antioxidants (lycopene, retinol, ascorbic acid, vitamin E, and selenium) are unlikely to causally associate with reduced epilepsy risk, except for ß-carotene.

The role of HIF-1α/HO-1 pathway in hippocampal neuronal ferroptosis in epilepsy.

Epilepsy, a common central nervous system disorder, remains an enigma in pathogenesis. Emerging consensus designates hippocampal neuronal injury as a cornerstone for epileptogenic foci, pivotal in epileptic genesis and progression. Ferroptosis, a regulated cell death modality hinging on iron, catalyzes lipid reactive oxygen species formation through iron and membrane polyunsaturated fatty acid interplay, culminating in oxidative cell death. This research investigates the role of hypoxia-inducible factor (HIF)-1α/heme oxygenase (HO)-1 in hippocampal neuron ferroptosis during epilepsy. Untargeted metabolomics exposes metabolite discrepancies between epilepsy patients and healthy individuals, unveiling escalated oxidative stress, heightened bilirubin, and augmented iron metabolism in epileptic blood. Enrichment analyses unveil active HIF-1 pathway in epileptic pathogenesis, reinforced by HIF-1α signaling perturbations in DisGeNET database. PTZ-kindled mice model confirms increased ferroptotic markers, oxidative stress, HIF-1α, and HO-1 in epilepsy. Study implicates HIF-1α/HO-1 potentially regulates hippocampal neuronal ferroptosis, iron metabolism, and oxidative stress, thereby promoting the propagation of epilepsy.

Causal relationship between addictive behaviors and epilepsy risk: A mendelian randomization study.

BACKGROUND: Previous studies have reported inconsistent results regarding the potential relationships between addictive behaviors and the risk of epilepsy. OBJECTIVE: To assess whether genetically predicted addictive behaviors are causally associated with the risk of epilepsy outcomes. METHODS: The causation between five addictive behaviors (including cigarettes per day, alcoholic drinks per week, tea intake, coffee intake, and lifetime cannabis use) and epilepsy was evaluated by using a two-sample Mendelian Randomization (MR) analysis. The inverse-variance weighted (IVW) method was used as the primary outcome. The other MR analysis methods (MR Egger, weighted median, simulation extrapolation corrected MR-Egger, and Mendelian Randomization Pleiotropy Residual Sum and Outlier (MR-PRESSO)) were performed to complement IVW. In addition, the robustness of the MR analysis results was assessed by leave-one-out analysis. RESULTS: The IVW analysis method indicated an approximately 20% increased risk of epilepsy per standard deviation increase in lifetime cannabis use (odds ratio [OR], 1.20; 95% confidence interval [CI]), 1.02-1.42, P = 0.028). However, there is no causal association between the other four addictive behaviors and the risk of epilepsy (cigarettes per day: OR, 1.04; 95% CI, 0.92-1.18, P = 0.53; alcoholic drinks per week: OR, 1.31; 95% CI, 0.93-1.84, P = 0.13; tea intake: OR, 1.15; 95% CI, 0.84-1.56, P = 0.39; coffee intake: OR, 0.86; 95% CI, 0.59-1.23, P = 0.41). The other MR analysis methods and further leave-one-out sensitivity analysis suggested the results were robust. CONCLUSION: This MR study indicated a potential genetically predicted causal association between lifetime cannabis use and higher risk of epilepsy. As for the other four addictive behaviors, no evidence of a causal relationship with the risk of epilepsy was found in this study.

The Negative Allosteric Modulator EU1794-4 Reduces Single-Channel Conductance and Ca2+ Permeability of GluN1/GluN2A N-Methyl-d-Aspartate Receptors.

NMDA receptors are ligand-gated ion channels that mediate a slow, Ca2+-permeable component of excitatory synaptic currents. These receptors are involved in several important brain functions, including learning and memory, and have also been implicated in neuropathological conditions and acute central nervous system injury, which has driven therapeutic interest in their modulation. The EU1794 series of positive and negative allosteric modulators of NMDA receptors has structural determinants of action near the preM1 helix that is involved in channel gating. Here, we describe the effects of the negative allosteric modulator EU1794-4 on GluN1/GluN2A channels studied in excised outside-out patches. Coapplication of EU1794-4 with a maximally effective concentration of glutamate and glycine increases the fraction of time the channel is open by nearly 1.5-fold, yet reduces single-channel conductance by increasing access of the channel to several subconductance levels, which has the net overall effect of reducing the macroscopic current. The lack of voltage-dependence of negative modulation suggests this is unrelated to a channel block mechanism. As seen with other NMDA receptor modulators that reduce channel conductance, EU1794-4 also reduces the Ca2+ permeability relative to monovalent cations of GluN1/GluN2A receptors. We conclude that EU1794-4 is a prototype for a new class of NMDA receptor negative allosteric modulators that reduce both the overall current that flows after receptor activation and the flux of Ca2+ ion relative to monovalent cations. SIGNIFICANCE STATEMENT: NMDA receptors are implicated in many neurological conditions but are challenging to target given their ubiquitous expression. Several newly identified properties of the negative allosteric modulator EU1794-4, including reducing Ca2+ flux through NMDA receptors and attenuating channel conductance, explain why this modulator reduces but does not eliminate NMDA receptor function. A modulator with these properties could have therapeutic advantages for indications in which attenuation of NMDA receptor function is beneficial, such as neurodegenerative disease and acute injury.

Genetic Interaction of H19 and TGFBR1 Polymorphisms with Risk of Epilepsy in a Chinese Population.

PURPOSE: Long non-coding RNA H19 was highly expressed in the latent period of epilepsy, contributing to apoptosis of hippocampal neurons by targeting let-7b. Transforming growth factor beta receptor 1 (TGFBR1), a target of let-7b, is located on the susceptibility locus for epilepsy. In this context, we investigated the association between tagSNPs in long non-coding RNA H19 and transforming growth factor beta receptor 1 (TGFBR1) rs6478974 and the risk of epilepsy. PATIENTS AND METHODS: The present study consisted of 302 patients with epilepsy and 612 age- and gender-matched controls. The polymorphisms were analyzed using a TaqMan allelic genotyping assay. H19 and TGFBR1 mRNA levels were determined using quantitative real-time polymerase chain reaction. RESULTS: The TGFBR1 AT and TT genotypes emerged as a protective factor for the risk of epilepsy (AT vs AA: adjusted OR = 0.59, 95% CI: 0.39-0.89, P = 0.01; TT vs AA: adjusted OR = 0.53, 95% CI: 0.35-0.80, P = 0.002, respectively). The protective effect was also observed in recessive genetic model (adjusted OR = 0.56, 95% CI: 0.38-0.82, P = 0.003). Individuals carrying the rs6478974 TT genotype had lower levels of TGFBR1 mRNA. Moreover, the TCTAT and TCCAA haplotypes emerged as a risk factor for epilepsy and the rs3741219-rs2839698-rs6478974 was associated with an interactive effect on the risk of epilepsy. CONCLUSION: The current study provides evidence of the rs6478974 TT genotype decreasing the susceptibility to epilepsy by reducing the levels of TGFBR1 mRNA.

Immune-mediated epilepsy with GAD65 antibodies.

Anti-GAD65 antibodies have been identified in both acute/subacute seizures (limbic encephalitis and extralimbic encephalitis) and chronic isolated epilepsy. The evidence of high serum titers and intrathecal synthesis play a fundamental role in diagnosis but poorly correlate with disease severity or response to therapies. It remains controversial whether anti-GAD65 Abs are the pathogenic entity or only serve as a surrogate marker for autoimmune disorders mediated by cytotoxic T cells. Unlike other immune-mediated epilepsy, although multiple combinations of therapeutics are used, the efficacy and prognosis of patients with GAD65-epilepsy patients are poor. Besides, GAD65-epilepsy is more prone to relapse and potentially evolve into a more widespread CNS inflammatory disorder. This article reviews the recent advances of GAD65-epilepsy, focusing on the diagnosis, epidemiology, pathophysiology, clinical features, and treatment, to better promote the recognition and provide proper therapy for this condition.

Lentivirus-mediated microRNA-26a-modified neural stem cells improve brain injury in rats with cerebral palsy.

This study is launched to investigate the effect of lentivirus-mediated microRNA-26a (miR-26a)-modified neural stem cells (NSCs) in brain injury in rats with cerebral palsy (CP). The successfully constructed miR-26a lentivirus expression vector and empty vector virus were used to modify NSCs. The model of CP with ischemia and anoxia was established in rats. NSCs and miR-26a-NSCs were stereoscopically injected into the cerebral cortex of the modeled rats, respectively. The survival and migration of NSCs infected with recombinant lentivirus expressing green fluorescence in vivo was observed under a light microscope. The neurobehavioral functions, morphology, and ultrastructure of cerebral cortex and hippocampus, apoptosis of brain cells, expression of apoptosis-related protein caspase-3 and Bax, together with the expression of the glial fibrillary acidic protein (GFAP) in cerebral cortex and hippocampus were determined. Expression of miR-26a in NSCs infected with plVTHM-miR-26a increased significantly. After NSCs transplantation, the neurobehavioral status of CP rats was improved, the degree of brain pathological injury was alleviated, the apoptotic index of cells in cerebral cortex and hippocampus and the expression of the apoptotic protein (caspase-3 and Bax) were decreased, the expression of GFAP were significantly decreased. After miR-26a-NSCs transplantation, these aforementioned results further improved or decreased. Our study suggests that miR-26a-modified NSCs mediated by lentivirus can improve brain injury, inhibit apoptosis of brain cells and activation of astrocytes in CP rats.

Association of genetic polymorphisms in CASP7 with risk of ischaemic stroke.

Caspase 7 (CASP7) is located on chromosome 10q25.3 that has been identified to be a susceptibility locus of ischaemic stroke (IS) by genome-wide association study. Elevated CASP7 was observed in IS, acting as a key apoptotic mediator in the development of IS. The aim of this study was to investigate the association between genetic polymorphisms in CASP7 and risk of IS. The CASP7 polymorphisms were genotyped using a TaqMan allelic discrimination assay. The expression levels of CASP7 mRNA were examined using quantitative polymerase chain reaction and luciferase activity was analyzed using the Dual Luciferase reporter assay. The rs12415607 in the promoter of CASP7 was associated with a reduced risk of IS (AA vs. CC: adjusted OR = 0.55, 95% CI: 0.38-0.80, P = 0.002; CA/AA vs. CC: adjusted OR = 0.70, 95% CI: 0.54-0.91, P = 0.007; AA vs. CC/CA: adjusted OR = 0.64, 95% CI: 0.46-0.90, P = 0.01; A vs. C: adjusted OR = 0.74, 95% CI: 0.62-0.89, P = 0.001). Moreover, the rs12415607 AA genotype carriers exhibited lower levels of CASP7 mRNA and the rs12415607 A allele decreased the promoter activity. These findings indicate that the rs12415607 A allele induces lower levels of transcriptional activity and CASP7 mRNA, and thus is associated with a reduced risk of IS.

circRNA/lncRNA-miRNA-mRNA Network in Oxidized, Low-Density, Lipoprotein-Induced Foam Cells.

Although long noncoding RNAs (lncRNAs) and circular RNAs (circRNAs) have been suggested to play important roles in the pathogenesis of diseases, atherosclerosis-related lncRNAs and circRNAs remain rarely reported. This study aimed to explore the underlying molecular mechanisms of atherosclerosis based on the competing endogenous RNA (ceRNA) regulatory hypothesis of lncRNAs and circRNAs. The expression profiles of circRNAs, lncRNAs, and mRNAs in human THP-1 macrophages treated with oxidized low-density lipoprotein (an in vitro atherosclerosis model), or not, were obtained from the Gene Expression Omnibus database under accession numbers GSE107522, GSE54666, and GSE54039, respectively. The present study identified 29 differentially expressed circRNAs in GSE107522, 544 differentially expressed genes (DEGs) in GSE54666, and 502 DEGs and 231 differentially expressed lncRNAs in GSE54039 datasets by using the Linear Models for Microarray Data method. Eight DEGs were found to be shared and expressed with the consistent trend in GSE54666 and GSE54039 datasets. Two of them (ASPH, aspartate beta-hydroxylase; and PDE3B, phosphodiesterase 3B) were suggested to be crucial based on functional enrichment, protein-protein interaction, and ceRNA network analyses. ASPH, through interaction with CACNA2D4 (calcium voltage-gated channel auxiliary subunit alpha2delta 4), may be associated with atherosclerosis by regulating the cellular response to calcium ion; and PDE3B may exert roles in negative regulation of angiogenesis through cross talk with ELMO1 (engulfment and cell motility 1). Furthermore, the expression of ASPH and PDE3B may be regulated by hsa_circ_0028198/hsa_circ_0092317/XIST-miR-543; PDE3B expression may be also modulated by hsa_circ_0092317/hsa_circ_0003546/H19/XIST-miR-326. In conclusion, our identified ceRNA interaction axes may possibly be important targets for treatment of atherosclerosis.

Association between MEG3/miR-181b polymorphisms and risk of ischemic stroke.

BACKGROUND: Recent evidence suggests that long non-coding RNAs (lncRNAs) are key regulators in the pathological process of ischemic stroke (IS). Maternally expressed gene 3 (MEG3) was observed to be up-regulated in IS, acting as a competing endogenous RNA for miR-181b to regulate ischemic brain injury. The purpose of this study was to evaluate the association of tagSNPs in MEG3 (i.e., rs7158663 and rs4081134) and miR-181b rs322931 with IS risk. METHODS: Genomic DNA was extracted from blood samples of 509 patients with IS and 668 healthy controls. Genotyping of MEG3 rs7158663, rs4081134, and miR-181b rs322931 was performed by TaqMan assay. The transcriptional activity was measured using the Dual-Luciferase Reporter Assay kit. RESULTS: Single-site analysis revealed a significantly higher risk of IS being associated with miR-181b rs322931 CT and CT/TT genotypes (CT vs. CC: adjusted OR = 1.48, 95% CI: 1.13-1.95, P = 0.005; CT/TT vs. CC: adjusted OR = 1.52, 95% CI: 1.17-1.97, P = 0.002). Combined analyses revealed that combined genotypes (rs7158663 GG + rs322931 CT/TT and rs7158663 AG/AA + rs322931 CT/TT) increased IS risk compared to genotypes of rs7158663 GG + rs322931 CC. Stratification analyses showed that patients carrying miR-181b rs322931 CT/TT genotypes had higher levels of low-density lipoprotein cholesterol (LDL_C) (P = 0.01). Moreover, results from logistic regression analysis showed that rs322931 CT/TT genotypes were risk factors besides hypertension, total cholesterol, triglyceride, and LDL_C. Further dual-luciferase reporter assay showed that the rs322931 T allele had lower levels of luciferase activity than the rs322931 C allele. CONCLUSION: These findings indicate that miR-181b rs322931 may singly or jointly contribute to the risk of IS.

A Functional Polymorphism rs145204276 in the Promoter of Long Noncoding RNA GAS5 Is Associated with an Increased Risk of Ischemic Stroke.

Long noncoding RNAs (lncRNAs) play crucial roles in the regulation of pathological process of ischemic stroke (IS) via affecting cell apoptosis, inflammation, cell death, and angiogenesis. LncRNA growth arrest-specific 5 (GAS5) was observed to be up-regulated in IS, acting as a competing endogenous RNA for miR-137 to mediate the Notch1 signaling pathway. In this study, we aimed to whether an insertion/deletion polymorphism (rs145204276) in the promoter of GAS5 was related to the risk of IS. The rs145204276 was genotyped using polymerase chain reaction (PCR)-polyacrylamide gel electrophoresis in 509 patients with IS and 668 healthy controls with frequencies matched to cases regarding age, gender, living area, and ethnicity. The GAS5 expression levels were determined using qPCR and relative luciferase activity was measured using the Dual Luciferase assay system. The presence of del/del genotype and del allele was associated with an increased risk of IS [del/del versus ins/ins: adjusted odds ratio (OR) = 2.06, 95% confidence interval (CI): 1.37-3.11; recessive model: adjusted OR = 2.07, 95% CI: 1.41-3.04; del versus ins: adjusted OR = 1.31, 95% CI: 1.08-1.57]. Results from logistic regression analysis identified risk factors for IS, including hypertension, total cholesterol, triglyceride, low-density lipoprotein cholesterol, and rs145204276 del/del genotype. Furthermore, patients carrying rs145204276 del/del genotype had significantly higher levels of GAS5 and cells transfected with rs145204276 del allele exhibited a larger increase in luciferase activity. These findings indicate that rs145204276 del allele exhibited a significant association with an increased IS susceptibility by elevating the transcriptional activity and subsequently enhancing the expression of lncRNA GAS5.

Proton magnetic resonance spectroscopy and cognitive impairment in patients with ischemic white matter lesions.

BACKGROUND: The purpose of this study is to investigate the relationship between the cognitive impairment and NAA/Cr and Cho/Cr ratios in the proton magnetic resonance spectroscopy ((1)HMRS), and to assess the importance of (1)HMRS in the early diagnosis of cognitive impairment in patients with ischemic white matter lesions (WMLs). MATERIALS AND METHODS: A total of 45 patients (23 males and 22 females) with the ischemic WML were divided into mild WML group (n = 15), moderate WML group (n = 15), and severe WML group (n = 15). A total of 15 healthy controls (8 males and 7 females) with no WML on magnetic resonance imaging were included. (1)HMRS focusing on the frontal lobe white matter around the anterior horn of the lateral ventricle and Montreal Cognitive Assessment (MoCA) were conducted. RESULTS: Patients with more severe WML had lower MoCA scores. The NAA/Cr ratio in (1)HMRS was reduced in all the patients and was strongly correlated with the total MoCA scores (r = 0.845, P < 0.001). The Cho/Cr ratio in (1)HMRS was increased in mild and moderate patients, was negatively correlated with the total MoCA scores (r = 0.907, P < 0.001). The Cho/Cr ratio was reduced in the severe patients and was positively correlated with the total MoCA scores (r = 0.937, P < 0.001). In addition, NAA/Cr and Cho/Cr ratios in (1)HMRS were changed in patients with the mild WML whose total MoCA scores were similar to the controls. CONCLUSION: Our results suggest that NAA/Cr and Cho/Cr ratios in (1)HMRS are useful indicators for early diagnosis of ischemic WML and cognitive impairment in patients with ischemic WML.

Hemichorea associated with non-ketotic hyperglycaemia: a case report.

Hemichorea is a rare complication of non-ketotic hyperglycaemia. Patients with this syndrome have classically longstanding, poorly controlled diabetes. Here, we report a patient presenting hemichorea without diabetic history. His symptom resolved rapidly after correction of hyperglycaemia. We suggest that hemichorea may be the first manifestation of undiagnosed diabetes.