European Journal of Pharmacology

Recurrent hypoglycemia increases cognitive impairment in diabetes mellitus patients. Following cerebral neuron injury, endothelial cells provide morphological, metabolic and immune support to damaged neurons. We investigated the inflammatory mechanism involved in hippocampal neuron degeneration. Behavioral experiments, including the open field test (OFT) and the Morris water maze test, were performed to measure cognitive changes. Using a vascular ring experiment, we evaluated vasodilation of the carotid artery. ZBP1 expression was knocked down after transfection with small interfering RNA in a brain endothelial cell line (bEnd3). In this study, PANoptosis, a recently defined form of programmed cell death (PCD), was found to be increased by hypoglycemia in the hippocampus of type 2 diabetic mice in vivo and by low glucose in bEnd3 cells in vitro. ZBP1 knockdown decreased PANoptosis induced by low-glucose stimulation in high-glucose-cultivated bEnd3 cells. RNA transcriptomics sequencing revealed that AGE-RAGE signaling significantly changed after ZBP1 was knocked down in bEnd3 cells. Corresponding biochemical data confirmed that ZBP1 knockdown regulated the advanced glycation end products (AGEs)-Receptor for Advanced Glycation End Products (RAGE) axis in bEnd3 cells. We present the first evidence that hypoglycemia impaired cognition in mice with type 2 diabetes by activating brain endothelial ZBP1-mediated PANoptosis via the AGE-RAGE axis. ARTICLE HIGHLIGHTSO_LIPANoptosis, a newly defined form of programmed cell death, is induced in the hippocampus after recurrent hypoglycemia in male db/db mice. C_LIO_LIZBP1, a sensor of the PANoptosome, was activated in low glucose cultured brain endothelial cells. C_LIO_LIHypoglycemia impairs vasodilation and cognitive function in type 2 diabetic mice. C_LIO_LIOur study indicates that inhibiting ZBP1-PANoptosis and the AGE-RAGE axis may be a potential approach to prevent hypoglycemia-induced cognitive degeneration in individuals with type 2 diabetes. C_LI

BackgroundDeep brain stimulation (DBS) is a treatment option for Parkinson disease (PD). However, the effect of DBS on the arterial pressure (AP) remains unexplored. We aimed to develop an artificial baroreflex system for treating orthostatic hypotension (OH) due to central baroreflex failure in patients with PD. To achieve this, we developed an appropriate algorithm after estimating the dynamic responses of the AP to DBS using a white noise system identification method. MethodsWe randomly performed DBS while measuring the AP tonometrically in 3 trials involving 3 patients with PD treated with DBS. We calculated the frequency response of the AP to the DBS using a fast Fourier transform algorithm. Finally, the feedback correction factors were determined via numerical simulation. ResultsThe frequency responses of the systolic AP to random DBS were identifiable in all 3 trials, and the steady state gain was 8.24 mmHg/STM. Based on these results, the proportional correction factor was set to 0.12, and the integral correction factor was set to 0.018. The computer simulation revealed that the system could quickly and effectively attenuate a sudden AP drop induced by external disturbances such as head-up tilting. ConclusionAn artificial baroreflex system with DBS may be a novel therapeutic approach for OH caused by central baroreflex failure.

The neurosteroid allopregnanolone is a positive allosteric modulator of GABA(A) receptors, which has proved beneficial in the treatment of major depressive disorder and epilepsies. It also has a role in treating the mood swings that are associated with fluctuations in its level during the menstrual cycle. Nonetheless, a subset of women do not tolerate high levels of allopregnanolone. Iso-allopregnanolone, a negative allosteric modulator, as well as synthetic steroid antagonists are used to treat such conditions. However, steroid-based medications are difficult to deliver and their specificity of action can be unclear. Recently introduced novel nonsteroidal agents that, like iso-allopregnanolone, can reverse the action of positive allosteric modulators without changing the positive action of GABA, might provide an alternative. We surveyed a number of them on human 1{beta}3{delta} GABAARs using a [3H]muscimol binding assay. A 6-membered ring spiro-hydantoin, DKD99, allosterically reversed the positive allosteric action of allopregnanolone over a wide concentration range (6 to 1,000 nM). DKD99 shifted allopregnanolones modulation curve 10-fold to the right. Furthermore, it has a much lower affinity when exerting similar actions on 1{beta}3{gamma}2 receptors. Agents such as this have utility for elucidating underlying mechanisms and may offer an alternative pathway for the development of nonsteroidal therapies against the positive allosteric modulatory actions of neurosteroids.

Endogenous circadian oscillators regulate learning, cognitive performance and memory are disrupted due to circadian shifts. High-fat-high-fructose (H) diet, photoperiodic shifts induced chronodisruption (CD) and a combination (HCD) causes neurobehavioral perturbations wherein; the merits of exogenous melatonin in alleviating the said behavioral deficits are studied herein. Indices of anxiety (marble burying test, elevated plus maze test and hole board test) and depressive behavior (sucrose preference test, forced swim test and tail suspension test) were elevated in H, CD and HCD groups. Significant increments in the titres of thyroid hormone levels (T3, T4 and TSH) and mRNA levels of hippocampal pro-inflammatory genes (Tnf-, Il-1{beta}, Il-4, Il-6, Il-10, Il-12, Il-17, Mcp-1 and Nf-{kappa}b) in the said experimental groups corroborates with the said changes. Exogenous melatonin treatment to the said experimental groups viz. HM, CDM and HCDM; accounted for moderate to significant improvement in the said neurobehavioral perturbations and hippocampal inflammatory markers. Hippocampal BDNF-TrkB pathway genes of H, CD and HCD had recorded a non-significant downregulation in mRNA but without prominent changes in proteins. Likewise, melatonin-treated groups showed moderate to significant improvement in transcripts of Bdnf, Trkb, Nt-3, Nt-4, Psd-95 and Syn-1. Herein, we report neurobehavioral perturbations caused by a combination of H and CD. Melatonin-mediated improvement in neurobehavior and the corrective changes in hippocampal BDNF-TrkB pathway implies towards the potential anxiolytic and anti-depressive activity as reported herein.

Parkinsons disease (PD) is the second most progressive degenerative disorder of the brain due to dopaminergic (DA) neuron degenerations and alpha-synuclein (-Syn) accumulations. At present, the disease has no effective treatment. Therefore, the current study objective is to identify a novel anti-PD formula (Zhi-Shi-Huang-Wu Formula, F-2) computed at 8:4:2:1 ratio from HSP 70 promoter activators Valeriana jatamansi (V), Acori talarinowii (A), Scutellaria baicalensis (S), Fructus Schisandrae (F). Traditionally, V is used to cure memory impairments, A treats mental disorders, and chronic mild stress, S for neuroprotection, and F showed multiple therapeutic actions to treat insomnia. This study investigated the neuroprotective potential of the V, A, S, F, formula F-2 and its underlying molecular mechanisms in transgenic Caenorhabditis elegans models. A, S, F, and F-2 successfully restored 6-hydroxydopamine intoxicated DA neuron degenerations, reduced food-sensing behavior disabilities, and attenuated -Syn aggregations. Moreover, activates the lipid deposition and proteasome expressions to confirm -Syn degradations at the cellular level. Reactive oxygen species (ROS) cause oxidative stress, and A, S, F, and F-2 repressed ROS and raised SOD-3 expressions. Overall, these data indicate that V, A, S, F combined into F-2 (22.3%) are more effective against PD progression-like symptom than individual drugs V (0.7%), A (11.4%), S (9.6%), and F (12.6%). These improved neuroprotective actions of F-2 possibly due to following the antioxidative pathway. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=144 SRC="FIGDIR/small/709540v1_ufig1.gif" ALT="Figure 1"> View larger version (47K): org.highwire.dtl.DTLVardef@1a6f1f7org.highwire.dtl.DTLVardef@157a270org.highwire.dtl.DTLVardef@69a238org.highwire.dtl.DTLVardef@1194b5e_HPS_FORMAT_FIGEXP M_FIG C_FIG

BackgroundThe primary cause of death associated with opioids is opioid-induced respiratory depression (OIRD). Naloxone is used to reverse OIRD, but this drug is a competitive antagonist of {micro}-opioid receptor (MOR) and reverses analgesia, which limits its therapeutic use. Alternative non-opioid receptor antagonist-based approaches to OIRD treatment and prevention are needed. The aim of this study was to evaluate if setmelanotide (SET) is capable of reversing OIRD in a mouse model. MethodsC57BL/6J male and female mice and Sprague-Dawley rats were given IP morphine or fentanyl and then treated 15 min later with either SET or vehicle VEH (IP) in a random order. Breathing was recorded by barometric plethysmography, and pain sensitivity was measured by the tail-flick test. ResultsIn mice with OIRD, SET induced a 3-fold reduction of the apnea index, and decreased apnea duration as compared to the VEH treatment. SET increased respiratory rate and did not affect opioid-induced analgesia. Photostimulation of MC4R+ ChR2-expressing fibers in the parafacial region of MC4R-Cre mice elicited short-latency excitatory postsynaptic current in rostral ventral respiratory group (rVRG) pre-motoneurons projecting to the phrenic nucleus in the C3-C4 ventral horns of the spinal cord. Fentanyl inhibited the activity of rVRG neurons and SET reversed this effect. ConclusionsSET effectively treated OIRD by increasing respiratory rate and inducing a significant decrease in the number of apneas without decreasing analgesia.

{beta}-Carotene is a strong antioxidant and immunomodulator, but it breaks down in the presence of oxygen and isnt readily bioavailable. Nutrishield(R) {beta}-Carotene is a special starch-based microencapsulation system designed to protect {beta}-carotene from oxidation, make it easier to spread in the gastrointestinal environment, and improve its effectiveness. This study reported Nutrishield(R) as a formulation platform by using {beta}-carotene as a model compound. We compared how well it worked in vitro and in vivo with {beta}-carotene that wasnt encapsulated. We randomly put thirty-two male BALB/c mice into four groups (n=8): Control, LPS, LPS + {beta}-Carotene (10 mg/kg/day), and LPS + Nutrishield(R) (the same dose of {beta}-Carotene). After an intraperitoneal injection of LPS (1 mg/kg, twice weekly), the treatments were administered orally for 28 days. LPS administration significantly elevated TNF- (142 {+/-} 8 pg/mL), IL-6 (118 {+/-} 7 pg/mL), and serum 8-OHdG (5.4 {+/-} 0.3 ng/mL) in comparison to controls (TNF- = 62 {+/-} 5 pg/mL; IL-6 = 51 {+/-} 4 pg/mL; 8-OHdG = 2.1 {+/-} 0.2 ng/mL). Nutrishield(R) {beta}-Carotene supplementation significantly normalized cytokine profiles (TNF- = 69 {+/-} 4 pg/mL; IL-6 = 56 {+/-} 3 pg/mL; IL-10 increased from 38 {+/-} 2 to 76 {+/-} 3 pg/mL; p < 0.001 vs. LPS). Histological analysis demonstrated reduced hepatocellular vacuolation and renal tubular degeneration in the Nutrishield(R) {beta}-Carotene group relative to the {beta}-carotene and LPS groups. These findings demonstrate that Nutrishield(R) microencapsulation significantly enhances the functional antioxidant and anti-inflammatory efficacy of {beta}-carotene in a murine model of systemic inflammation. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=134 SRC="FIGDIR/small/711625v1_ufig1.gif" ALT="Figure 1"> View larger version (48K): org.highwire.dtl.DTLVardef@1da9828org.highwire.dtl.DTLVardef@3b8ef2org.highwire.dtl.DTLVardef@25e339org.highwire.dtl.DTLVardef@168e793_HPS_FORMAT_FIGEXP M_FIG C_FIG

BackgroundAlcohol-induced osteonecrosis of the femoral head (AIONFH) is an orthopedic disorder from chronic alcohol abuse, characterized by disrupted femoral head blood supply, osteocyte death and structural collapse. Current hip-preserving therapy is unsatisfactory, and most patients eventually require total hip arthroplasty. Panax Notoginseng Saponins (PNS), the core active component of Panax notoginseng, exerts pro-angiogenic and anti-osteocyte apoptosis effects, but its specific therapeutic mechanism remains unclear. ObjectiveThis study used network pharmacology, molecular dynamics simulation and animal experiments to identify PNSs active components, core targets and key pathways for AIONFH, verify its in vivo efficacy, and provide a scientific basis for clinical application. MethodsPNS active components, their targets and AIONFH-related targets were screened from databases; intersection targets constructed an interaction network, core targets were screened by three machine learning algorithms, with concurrent GO and KEGG analysis. Molecular docking was performed between core targets and PNS components; Gromacs 2022 conducted 100 ns simulation to evaluate complex stability. AIONFH rat models were grouped with 4-week intragastric intervention; pathology, immunofluorescence and PCR were used for detection. Results and DiscussionNetwork pharmacology identified 127 PNS targets and 18 intersections with 672 AIONFH targets. Six core targets (including FGF2, HSD11B1) were screened; KEGG indicated VEGF pathway as key. Ginsenoside Re bound HSD11B1 with the lowest binding energy (-12.4 kcal/mol), and 100 ns simulation confirmed complex stability. Animal experiments showed PNS improved trabecular structure and regulated osteocyte activity. PNS treats AIONFH via multi-component, multi-target mode, core mechanism being osteocyte apoptosis inhibition. Results and DiscussionNetwork pharmacology screening identified 127 potential targets of PNS, and 18 potential intersection targets were obtained by overlapping with 672 AIONFH-related targets. Six core targets including FGF2 and HSD11B1 were screened out by machine learning, and KEGG analysis indicated that the VEGF pathway and other pathways were the key signaling pathways for PNS action. Molecular docking showed that Ginsenoside Re had the lowest binding energy with HSD11B1 (-12.4 kcal/mol), and 100 ns molecular dynamics simulation confirmed the stable conformation of this complex. Animal experiments demonstrated that PNS could improve trabecular bone structure and regulate osteocyte activity. In summary, PNS exerts a therapeutic effect on AIONFH through a multi-component, multi-target and multi-pathway mode, with the core mechanism of inhibiting osteocyte apoptosis.

IntroductionIntestinal constipation (IC) is a common and early non-motor symptom in Parkinsons disease (PD), impacting patients quality of life. In this context, the Mediterranean diet plays a fundamental role in managing IC. This study aimed to evaluate the effects of a nutritional education program based on the principles of the Mediterranean diet on IC in individuals with PD. MethodsThis is a randomized, controlled, single-center, parallel-group, single-blind clinical study to evaluate the effectiveness of a nutritional education program based on the Mediterranean diet for three months, with a delayed-start design, in people with PD and IC. Participants were randomly allocated (1:1 ratio) to two groups: early-start (intervention from baseline to the third month) and delayed-start (intervention from the third to the sixth month) nutritional counseling, with an initial in-person assessment and monthly remote assessments. Bowel habits, adherence to the Mediterranean diet, and clinical variables were analyzed. ResultsAfter six months, a significant increase in the frequency of weekly bowel movements was observed (Early-start: 2.91 to 4.14; Delayed-start: 2.68 to 4.18 bowel movements/week; p < 0.001), along with changes in stool consistency and improved adherence to the Mediterranean diet over time. However, no significant differences were detected between the groups. ConclusionA nutritional education program based on the principles of the Mediterranean diet was associated with improved bowel habits and dietary adherence over time. These results support that locally adapted, low-cost dietary counseling may represent a complementary approach to the treatment of IC in individuals with PD in non-Mediterranean settings.

Inflammation and oxidative stress (OS) are key to Parkinsons disease (PD). We performed a cross-dataset integrative transcriptomic analysis to identify OS- and inflammation-related hub genes persistently dysregulated in PD and to evaluate their response to nutrigenomic interventions using publicly available datasets. Four GEO datasets (GSE7621, GSE20141, GSE20146, GSE49036) were analysed to identify differentially expressed genes (DEGs), which were intersected with GeneCards OS-inflammation gene sets. Functional enrichment analyses, including gene ontology (GO), pathway over-representation analysis (ORA), and protein-protein interaction (PPI) analysis, were used to identify key pathways and hub genes. Gene-food bioactive compound (FBC) association was explored by integrating PD signatures with nutrigenomic profiles from NutriGenomeDB. We identified 183 DEGs in PD, enriched in synaptic, dopaminergic, OS, and inflammatory pathways. Intersection analysis yielded 26 OS-inflammation-related genes and 10 central regulators, including TH, DDC, SNCA, LRRK2, HSPB1, and HSPA1B. revealed opposing transcriptional patterns, with several FBCs suppressing stress-related genes and upregulating dopaminergic markers such as TH, GCH1, and DDC. Overall, this integrative analysis highlights OS-inflammation gene networks in PD and identifies candidate diet-gene interactions that warrant further experimental validation

BackgroundIt has been shown that n-3 polyunsaturated fatty acids (n-3 PUFA) of marine origin exert significant beneficial effects on myocardial infarction (MI); however, the underlying mechanisms remained unclear. Ceramides play a vital role in the regulation of energy metabolism, mitochondrial function, and apoptosis. Through the integration of clincial studies and animal experiments, this study aimed to determine whether n-3 PUFA improved myocardial function by modulating ceramide metabolism. MethodsIn a case-control study, 100 patients with AMI and 100 healthy pariticipants were enrolled to measure serum ceramide concentrations. Meanwhile, mice were randomly allocated into 4 groups and administrated to a 3-week intervention with n-3 PUFA in triglyceride and phospholipid forms. A mouse model of MI was then established, followed by an additional 4 weeks of continuous intervention. Subsequent comprehensive assessments of cardiac function were performed in the mice. Finally, the mice were euthanized to conduct targeted ceramide lipidomic analysis and other relevant assays. ResultsThe levels of serum C16:0-, C18:0-, C20:0-, C24:1-ceramides and total ceramides in patients with acute myocardial infarction (AMI) were significantly higher compared with the healthy controls. In the murine model of myocardial infarction, pathological analysis via TTC staining demonstrated that interventions with fish oil (triglyceride form) and krill oil (phospholipid form) both significantly reduced myocardial infarct size. Concomitant echocardiographic assessment confirmed that both treatments markedly elevated left ventricular ejection fraction (LVEF), with the magnitude of improvement being significantly superior to that of the model control group. Concurrently, compared with the model group, the concentrations of ceramides in cardiac tissue and serum were significantly lower in the groups with fish oil and krill oil intervention. Western blot analysis further confirmed that n-3 PUFA intervention upregulated adiponectin expression, reduced ceramide accumulation in myocardial tissue, and inhibited mitochondria-mediated cardiomyocyte apoptosis, thereby improving cardiac function and prognosis following myocardial infarction. ConclusionsThis work demonstrates that n-3 PUFA exert cardioprotective effects following MI mediated by adiponectin-ceramide axis. However, there is no significant difference regarding therapeutic efficacy of n-3 PUFA in triglyceride or phospholipid forms.

A hallmark of dilated cardiomyopathy (DCM) is calcium mishandling, including reduced transport activity of the SERCA calcium pump in cardiac muscle cells. This has focused attention on SERCA as mechanism of disease and potential therapeutic target. Previously, diminished SERCA activity has been attributed to decreased protein expression, but recent studies suggest SERCA levels are unchanged in DCM. Thus, another mechanism must be responsible for the deficit. Since proteolysis is increased and proteosome function is impaired in DCM, we reasoned that accumulation of toxic protein fragments may contribute to SERCA dysfunction. In particular, previous studies showed diverse species of hydrophobic -helices can inhibit SERCA, so we hypothesized that SERCA may become congested with transmembrane peptides that mimic endogenous regulatory partners. We purified cell membranes from non-failing and DCM human ventricles and subjected them to mass spectrometry to identify protein species upregulated in DCM. Select candidates were screened for binding and inhibition of SERCA. Several small membrane proteins and membrane protein fragments bound avidly to SERCA and significantly reduced cellular calcium stores. The data suggest a novel pathophysiological mechanism in which transmembrane protein debris obstructs SERCA function and regulation, contributing to cardiac muscle dysfunction in heart failure.

Inflammatory bowel disease (IBD) is characterised by chronic pain, a debilitating symptom for which effective treatments are few and far between. IBD pathogenesis includes the prevalence of a variety of pro-inflammatory cytokines, including the Interleukin-6 (IL-6) family members Il-6 and Oncostatin M (OSM). Previous research has shown disruption of OSM signaling can modulate nociceptor sensitization and activation, however the downstream signalling pathway is unknown. When an in silico analysis of murine colonic sensory neuronal populations was undertaken for receptor expression for OSM and other factors necessary for intracellular signaling, we can find diverse expression indicative of functional signaling. We were able to observe that hyper Il-6 (Il-6 bound to the soluble Il-6 receptor) and OSM can elicit activation of a subset of murine sensory neurons by finding an increase in calcium mobilization following superfusion. This could then be attenuated by the pharmacologic inhibition of all janus kinases or interestingly, TYK2 alone. Furthermore, inhibition of transient receptor potential vanilloid 1 or transient receptor potential ankyrin 1 ion channels, which are known to be sensitized by OSM in other sensory neurons also reduced the proportion of OSM-responsive neurons. This further understanding of OSM signaling in sensory neurons creates avenues for more extensive research into the molecular mechanisms occurring as well as the potential to exploit these therapeutically to induce analgesia in a subset of neurons.

Background: Clinical evidence on the contemporary management and functional outcomes of patients with Wernicke encephalopathy remains limited. This study aimed to clarify the nationwide patterns of thiamine administration and functional outcomes at discharge. Methods: Using the Japanese nationwide inpatient Diagnosis Procedure Combination database, we identified patients hospitalized with Wernicke encephalopathy between July 2010 and March 2024. Initial intravenous thiamine doses were categorized as low ([&le;]300 mg/day), medium (301-900 mg/day), or high (>900 mg/day). Outcomes included in-hospital mortality and functional status (Barthel Index) at discharge. Results: We identified 7856 patients with Wernicke encephalopathy. Over the 13-year study period, the proportion of patients receiving initial high-dose thiamine increased markedly from 5.4% to 49.0%, while the frequency of low-dose therapy decreased from 83.0% to 37.9%. Despite prompt intervention [median time to initial administration: 0 days (interquartile range, 0 to 0 days)], 56.1% of patients were discharged with impaired activities of daily living (Barthel Index <90), and the in-hospital mortality rate was 3.8%. Conclusions: High-dose thiamine treatment is increasingly implemented for Wernicke encephalopathy in Japan. Although in-hospital mortality was relatively low, the high prevalence of functional impairment at discharge, despite early treatment initiation, indicates substantial burden of Wernicke encephalopathy. Given the limited clinical evidence, further research investigating the optimal thiamine dose and develop effective primary prevention strategies for Wernicke encephalopathy is needed.

Temozolomide is the gold standard chemotherapeutic agent used in the treatment of glioblastoma multiforme. Yet its pharmacological use has been linked to the emergence of depressive- and/or anxiety-like behaviors, probably through the inhibition of hippocampal neurogenesis. Since prior studies reporting these negative effects were based on prolonged treatment paradigms (i.e., from 2 weeks to up to 6 months), and given the few reports that have included female rodents in their studies, our approach aimed at further characterizing the behavioral effects induced by temozolomide (25 mg/kg, 1 or 2 cycles, 5 days/cycle) in a mixed-sex cohort of adult rats. To do so, rats were scored across time through specific behavioral tests that capture diverse manifestations of affective-like responses (forced-swim, open field, novelty-suppressed feeding and sucrose preference) or cognitive performance (Barnes maze). At the neurochemical level, we ascertained the effects of 2 cycles of temozolomide on hippocampal neurogenesis (neural progenitors with NeuroD) and other potential neuroplasticity targets (i.e., FADD, BDNF). The main results showed that temozolomide induced unexpected antidepressant-like responses in a treatment-duration manner while decreased hippocampal FADD, a neuroplastic marker previously associated with the acute and repeated actions of most antidepressants. These results break the prior dogma linking increased hippocampal neurogenesis with antidepressant-like efficacy, and suggest that other mechanisms of action, such as the one described through the neuroplastic molecule FADD, might be responsible for the antidepressant-like actions of temozolomide, even in the presence of impaired neurogenesis. Our results, in conjunction with the prior data, suggested cycle- and/or length-dependent treatment effects in terms of temozolomides antidepressant- vs. depressant-like profile, while proposing a novel biomarker of its treatment response.

The Na+/K+-ATPase (NKA) regulates ion balance in the kidney and influences cellular processes like proliferation and apoptosis through its signal transduction. The endogenous ligand 20-Hydroxyeicosatetraenoic acid (20-HETE) contributes to inflammation and fibrosis in chronic kidney disease (CKD) and inhibits NKA activity in renal tubules. However, the molecular mechanism of this interaction remains unclear. In this study, we used in-silico approach to investigate the potential interaction between 20-HETE and NKA. Various ligands, including known NKA ligands such as cardiotonic steroids (CTS), 20-HETE, and negative controls, were docked using rigid and Induced Fit Docking to predict the affinity of the ligands toward NKA. Binding free energy calculations with the Prime Molecular mechanics with generalized Born and surface area (Prime MM/GBSA) tools were used to confirm the involvement of key amino acids in ligand-receptor interactions. The docking analyses revealed that 20-HETE exhibited a binding affinity comparable to negative control, with some differences between rigid and induced fit docking. Binding free energy data highlighted key amino acids in the 20-HETE and NKA interaction. Interaction fingerprint and mutations such as Ala330Gly and Val329Ala significantly reduced binding free energy, while Thr804Ala showed a notable decrease, underscoring the potential importance of these amino acids in ligand stabilization. These findings provide computational evidence supporting potential direct interaction between 20-HETE and NKA and identify candidate residues for future experimental validation.

Introduction Stroke is a leading cause of disability and death in adults with type 2 diabetes (T2D). We evaluated the comparative stroke risk in Veterans with T2D initiated on either of two glucose-lowering medications: GLP-1 receptor agonists (GLP-1RA) or SGLT-2 inhibitors (SGLT2i). Patients and Methods We conducted a retrospective cohort study on diabetic Veterans aged 40 and older with no prior history of stroke or transient ischemic attack, who started on a GLP-1RA or SGLT2i between 2014 and 2021. Patients with contraindications or prior exposure to medication were excluded. Using national Veteran health data, we identified 195,072 [SS1.1]eligible individuals and followed them from treatment initiation until stroke, death, loss to follow up, or end of follow up, whichever came first. Primary outcome was incident stroke, and secondary outcomes included ischemic and hemorrhagic stroke. We applied Kaplan-Meier methods and Cox proportional hazards models. Adjusted associations were estimated using inverse probability weighting. Results Both unadjusted and adjusted analyses suggest GLP-1RA users have reduced stroke incidence compared SGLT-2i users[HS2.1] (HR = 0.[HS3.1]67, 95% CI 0.64-0.69; HR = 0.72, 95% CI 0.69-0.75). Similar results were found in secondary outcome and stratified analyses, with GLP-1RA users having reduced stroke risk compared to SGLT2i users for all age groups, chronic kidney disease stages, and hemoglobin A1c levels. Discussion and Conclusion GLP-1RA treatment was associated with a lower risk of stroke compared with SGLT2i treatment in Veterans with T2D. These findings were consistent for ischemic and hemorrhagic strokes, suggesting potential differences in stroke risk between the treatments.

BACKGROUNGBisphenol A (BPA) has been linked to hypertension and disturbances in lipid metabolism; however, limited evidence is available regarding its association with hypertensive intracerebral hemorrhage (ICH). METHODSA multicenter, retrospective case-control study was conducted involving 129 participants, including individuals from an ICH group and healthy controls. Standard assays were employed to assess serum thyroid function, lipid profiles, serum fatty acid-binding [x]protein 4 (FABP4), oxidative stress markers, gap junction proteins, Wnt/{beta}-catenin signaling pathway activity, and expression changes of S100A8-mediated inflammatory cytokines involved in gut-brain interactions. Correlation analyses using Pearson and Spearman methods revealed that both BPA exposure and low T3 levels were significantly associated with elevated diastolic blood pressure, altered lipid metabolism, gut microbiota composition, and microglial activation. RESULTSGender-based disparities in lipid metabolism were identified. Changes in {beta}3-adrenergic receptor and neuromodulin-1 expression appear to influence fat regulation and attenuate oxidative stress responses. Subsequently, increased expression of gap junction proteins and activation of the Wnt/{beta}-catenin signaling pathway contribute to metabolic reprogramming and alterations in biochemical kinetics. Gut microbiota analysis demonstrated that, compared to controls, the ICH group exhibited significant dysbiosis and reduced alpha diversity. Further correlation analyses indicated that BPA levels were positively associated with FABP4 and oxidative stress markers, while S100A8 showed a strong dependence on microglial expression. CONCLUSIONThe interplay between lipid metabolism dysfunction and pro-inflammatory cytokines enhances vascular vulnerability. Collectively, BPA exposure, oxidative stress, and microglia-mediated neuroinflammation are significantly associated with an elevated risk of hypertensive ICH. China Clinical Trial Registry registration noticeFrom: China Clinical Trials Registry <chictr@vip.qq.com>+To:guopingwang60a<guopingwang60a@163.com> yunyanshuangfei <yunyanshuangfei@126.com> FUNDINGThis work was supported by the Natural Science Foundation of Shanxi Province (grant no. 201701D121177) Key informationGender-specific differences were observed in lipid metabolism and oxidative stress parameters; BPA exposure was shown to induce lipid metabolic disturbances, promote excessive production of oxidative stress byproducts, and consequently elevate oxidative stress responses; BPA was associated with stress-induced alterations in thyroid hormone function, further exacerbating dysregulation of lipid metabolism and oxidative stress; Fatty acid binding protein 4 (FABP4), a key adipokine implicated in metabolic disorders and adipose tissue inflammation, exhibited a significant positive correlation with serum BPA levels, whereas low levels of triiodothyronine (T3) were negatively correlated with FABP4. These findings suggest that serum FABP4 may serve as a biochemical marker for chronic low-grade adipose tissue inflammation and metabolic dysfunction; Gap junction proteins and the Wnt/{beta}-catenin signaling pathway may contribute to microglial activation and mediate neuroinflammatory responses, nerve injury, and secondary pathological processes in obesity-related cerebral hemorrhage.

Alzheimers disease (AD) is a common neurodegenerative disorder primarily caused by Amyloid-beta (A{beta}) toxicity. Therefore, there is an urgent need to develop novel, effective, and safe drugs to treat AD. Traditional Chinese Medicine (TCM) has a long history of use in protecting against memory impairments. Recently, TCM has attracted growing attention from researchers as a source of potent neuroprotective compounds. In this study, we focus on four TCM herbs with multiple therapeutic properties: Valeriana jatamansi (V; 20 mg/mL), Acori tatarinowii (A; 10 mg/mL), Fructus Schisandrae (F; 5 mg/mL), and Scutellaria baicalensis (S; 2.5 mg/mL). The aim is to develop a neuroprotective anti-AD formulation, named "Zhi-Shi-Wu-Huang" derived from V, A, F, and S, and evaluate its efficacy in transgenic Caenorhabditis elegans models of AD. These four TCM herbs are among the most potent activators of the HSP-70 promoter, promoting the expression of heat shock protein 70 (HSP-70), which helps prevent protein misfolding and aggregation. Additionally, V, A, F, S, and the Zhi-Shi-Wu-Huang formula were found to reduce reactive oxygen species (ROS) production and enhance the expression of superoxide dismutase-3 (sod-3) and chymotrypsin-like proteasomes. Our findings demonstrate that both the individual extracts (V, A, F, S) and the Zhi-Shi-Wu-Huang formulation significantly reduce A{beta}-induced toxicity in transgenic worms by activating the insulin/DAF-16 signaling pathway.

IntroductionInadequately controlled inflammation is a key driver of adverse cardiac remodelling after acute myocardial infarction (AMI). Central to this process is activation of the NLRP3 inflammasome-gasdermin D (GSDMD) pathway, which promotes pyroptosis and the release of the pro-inflammatory cytokine interleukin-1{beta} (IL-1{beta}), a mediator strongly associated with infarct severity and poor clinical outcomes. This study investigates whether repurposing the FDA-approved therapeutic Disulfiram, recently shown to inhibit GSDMD pore formation, could reduce inflammation and thus improve cardiac injury after AMI. Methods and ResultsCardiac ischemia-reperfusion (I/R) injury was induced in C57BL/6 mice by 60-minute ligation of the left coronary artery followed by reperfusion. Disulfiram (25 or 50 mg/kg) was administered at reperfusion and daily thereafter. Cardiac function was assessed by echocardiography, while fibrosis and inflammation were evaluated by histology, RT-PCR, immunohistochemistry and immunoblotting. Leukocyte populations in blood, spleen, bone marrow and heart were analysed by flow cytometry. In vitro, mouse bone marrow-derived macrophages (BMDMs) and PMA-differentiated THP-1 cells were treated with Disulfiram. Cytokine secretion, inflammatory gene expression and changes in cell viability (propidium iodide (PI) staining and lactate dehydrogenase (LDH) release) were measured. Disulfiram (50 mg/kg) significantly improved cardiac function 7 days post-I/R. This was accompanied by a significant reduction in cardiac fibrosis and inflammation, as reflected by a lower abundance of inflammatory cells in circulation and cardiac tissue. In LPS- and ATP/Nigericin-stimulated BMDMs and THP-1 cells, Disulfiram dose-dependently (0.1-50 {micro}M) reduced IL-1{beta} and IL-6 secretion and attenuated membrane permeability and cell lysis. ConclusionsThis study demonstrates that Disulfiram improves cardiac function post-AMI by ameliorating inflammation and fibrosis, which was associated with reductions in cytokine release from inflammatory cells in vitro. Therefore, targeting GSDMD by "repurposing" the FDA-approved drug, Disulfiram, may represent a novel way to provide cardio-protection post-AMI. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=82 SRC="FIGDIR/small/710794v1_ufig1.gif" ALT="Figure 1"> View larger version (11K): org.highwire.dtl.DTLVardef@7fd82eorg.highwire.dtl.DTLVardef@149de52org.highwire.dtl.DTLVardef@a2fe0eorg.highwire.dtl.DTLVardef@d56b4f_HPS_FORMAT_FIGEXP M_FIG C_FIG