Abstract The aim of this study was to investigate the protective effects of brown algae, namely Sargassum thunbergii (ST) and Sargassum fusiforme (SF), on memory and cognitive impairment, development of Alzheimer’s disease (AD), oxidative stress, and microglial activation in D-galactose (D-gal)-induced aging rats. Adult male Sprague Dawley rats were administered D-gal (150 mg/kg, i.p.) and a daily dose of hot water extract of ST (150 and 300 mg/kg) or SF (300 mg/kg) or phosphatidylserine [(PS) 30 mg/kg, positive control] for 13 weeks. ST, SF, and PS exhibited improved memory and cognition impairment in both radial arm maze and novel object recognition tests. Administration of ST, SF, and PS attenuated amyloid beta (Aβ) levels by decreasing Aβ production and increasing Aβ clearance-related proteins in the brains of D-gal-induced aging rats. However, only the ST group showed reduced expression of hyper-phosphorylated tau proteins in the brain by suppressing glycogen synthase kinase 3 beta (GSK3β) activities. Moreover, ST, SF, and PS also decreased acetylcholinesterase activity, oxidative stress, microglia activation, and inflammation, and increased the microglial M2 phenotype in the rat brain compared to D-gal-treated control rats. These results indicate that ST and SF could be potential candidates to ameliorate the risk of AD.

Oxidative stress can induce memory loss, which may be mitigated by antioxidants. Moringa leaves (Moringa oleifera) and butterfly pea flowers (Clitoria ternatea) possess flavonoid chemicals that function as natural antioxidants. This study seeks to evaluate the impact of combined Moringa leaf and butterfly pea flower infusions on enhancing the memory of mice and to identify the optimal dosage. The research utilized male white mice (Mus musculus), categorized into six groups: a normal group, a negative control (aquadest), a positive control (Ginkgo biloba), and three treatment groups receiving various infusion doses. The procedure had three phases: acclimatization (T0), intraperitoneal administration of hyoscine butylbromide (T1), and therapy (T2). The manufacture of INDUK BUNTEL (an infusion of moringa leaves and butterfly pea flowers) was conducted using an infusion extraction method. Testing on test subjects was conducted utilizing the Radial Arm Maze methodology. The findings indicated that the infusion combination at dose III (100 mg/KgBW: 100 mg/KgBW) was the most efficacious in enhancing the memory of mice. The mean mistake rate in mice was 16.44%, representing the lowest score among the treatment groups. The one-way ANOVA test indicated no significant difference across treatment groups with three distinct dose combinations (p > 0.05).

ABSTRACT Background and Objectives Stress in day-to-day life affects the body system differently by altering homeostasis and biological processes. The foremost sign of chronic stress-induced disorders is reflected in behavioral activity. Chronic stress affects the brain negatively, it modulates neurobehavioral activity and impairs the associated activities like learning, memory, and cognition. Emerging evidence suggests that virgin coconut oil (VCO), is an abundant source of natural antioxidants with anti-inflammatory, antidepressant, and neuroprotective potential. However, VCO's efficacy in ameliorating chronic restraint stress-induced abnormal changes is rarely understood. Hence, we aimed to evaluate VCO's neuroprotective potential in attenuating chronic restraint stress-induced neurobehavioral and biochemical alteration in rats. Methods A total of eighteen male Wistar rats were allocated into three groups (n = 6/group): Control (C) – saline (5 ml/kg/21 days), Stress (S) – restraint stress (6hrs/day/21 days), and Test (S + VCO) (6 hrs/day+5 ml/kg VCO for 21 days). The behavioral (Radial arm maze (RAM)) and biochemical (corticosterone (CORT)), reduced glutathione (GSH), and malonaldehyde (MDA) parameters were analyzed to determine the neuroprotective effect of VCO. Results The VCO-treated group has shown a significant increase in the percentage of correct choice entries and a reduction in error entries, highlighting an improvement in working and reference memory. Moreover, there was a substantial elevation in GSH, and a reduction in MDA and CORT levels, indicating a potent antioxidant activity of VCO. Additionally, significant body weight gain and reduced adrenal gland weight were noted in VCO-treated rats. Conclusions Our findings signify that VCO has therapeutic potential to improve spatial learning memory and antioxidant status in chronic restraint stress rats. Schematic depiction of the protective effect of virgin coconut oil on chronic restraint stress rats. Highlights VCO exhibits notable neuroprotection and convalesces the neuron. VCO reduces lipid peroxidation in the brain of chronic restraint stress rats. VCO retrieves spatial learning and memory in chronic restraint stress rats. Oral administration of VCO in rats prevents oxidative stress damage in the brain.

The olfactory bulb (OB) is an emerging neuroendocrine centre regulating appetite, metabolism, and behaviours such as those linked to anxiety, motivation and spatial navigation. These processes are likely mediated by one or more of the many hormone receptors found in the OB. For instance, recent studies show that selective OB deletion of the receptor for ghrelin and LEAP2 (GHSR; growth hormone secretagogue receptor) increases anxiety-like behaviour and impacts peripheral glucose and energy homeostasis. As GHSR function has been linked to motivated behaviours and spatial navigation, we decided to investigate whether OB-selective GHSR (OBGHSR) deletion affects motivation, using an operant progressive ratio schedule, and/or spatial navigation, using a Y maze and radial arm maze. In contrast to wild-type mice, our study shows that OBGHSR deletion increased motivated sucrose seeking after a short fast, but not in ad libitum fed mice, and had a mild effect to delay extinction learning. In both Y-maze and radial arm maze studies for spatial navigation, OBGHSR deletion reduced spatial exploration in terms of distance moved and arm entries. However, the proportion of correct and incorrect arm entries relative to the total number of entries was not affected in either the Y-maze or the radial arm maze, suggesting that spatial memory was not affected. Our study demonstrates that intact OBGHSRs in male mice normally restrain motivated sucrose seeking in a metabolic state-dependent manner and optimise spatial navigation by increasing exploration, without affecting spatial memory.

Background: Spatial memory impairment has significant negative influence on both survival and quality of life. Nigella sativa oil (NiSO) has been investigated for its potential to reduce memory impairments in various experimental models. Objectives: To assess the effects of NiSO on ketamine induced working and reference memory impairment in male Wistar rats. Methods: This experimental study was conducted from March 2020 to February 2021 in the Department of Physiology, after obtaining ethical approval from the Institutional Review Board of Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka, Bangladesh. For this purpose, 60 male Wistar rats were divided into normal memory (normal saline 5 ml/kg for 26 days), impaired memory (ketamine 15 mg/ kg during acquisition phase) and experimental (treated with NiSO 1 ml/kg for 26 days and ketamine 15 mg/kg during acquisition phase) groups. All groups underwent Radial arm maze (RAM) and Morris water maze (MWM) tests. Variables were working memory error (WME, re-entry into already visited arm) and reference memory error (RME, entry into non-baited arm) in RAM along with escape latency (EL, the moment of a rat’s entrance into the water upto it’s arrival at the platform) and target crossing (TC, number of crossing of the quadrant from where the platform was removed) in MWM. Data were expressed as mean±SEM and statistically analyzed with ANOVA followed by Bonferroni post-hoc test, where p≤0.05 was considered as significant. Results: Significantly (p≤0.001) higher WME and RME as well as significantly (p≤0.001) delayed EL and reduced TC were found in memory impaired rats when compared to normal memory rats. However, significantly (p≤0.001) lower WME and RME as well as significantly reduced EL (p≤0.001) and higher frequency of TC (p≤0.001) were observed in experimental rats, when compared to memory impaired rats. Moreover, these variables were almost similar in the experimental rats, in comparison to, those of normal memory rats except significantly (p≤0.01) higher TC in MWM test. Conclusions: NiSO prevented working and reference memory impairment as well as enhanced reference memory in rats. J Bangladesh Soc Physiol 2025;20(1): 19-31

Beta-sitosterol mitigates type 2 diabetes-induced cognitive deficits: a behavioral and molecular investigation in Swiss albino mice.

Memory impairment, characterized by reduced ability to recall facts, information, and experiences, is increasingly recognized as a major public health concern. This trend is largely driven by the rising prevalence of age-related cognitive decline and Alzheimer’s disease within the aging population. Aluminium chloride (AlCl₃), a well-established neurotoxicant, induces neurobehavioral and biochemical alterations that mimickey features of neurodegenerative disorders, thus serving as a reliable experimental model for evaluating neuroprotective agents. This study assessed the neuroprotective efficacy of quercetin in a rat model of AlCl₃-induced neurotoxicity. Fifty adult male rats (n = 10 per group) were randomly assigned into five groups. Neurotoxicity was induced by oral administration of AlCl₃ (100 mg/kg/day) for 14 days. Subsequently, from days 14 to 35, rats received daily treatments of quercetin (100 or 200 mg/kg), donepezil (3 mg/kg), or vehicle control. AlCl₃ exposure significantly impaired body weight gain, feed intake, locomotor activity, grooming behaviour, and spatial memory performance. Quercetin treatment markedly ameliorated these deficits, as evidenced by improved performance in Y-maze and radial-arm maze tasks. Biochemical analysis revealed that quercetin significantly reduced lipid peroxidation, enhanced total antioxidant capacity, and modulated inflammatory responses by decreasing pro-inflammatory cytokines (IL-1β, TNF-α) and elevating anti-inflammatory IL-10 levels. Furthermore, quercetin restored acetylcholine and brain-derived neurotrophic factor (BDNF) concentrations and preserved hippocampal cytoarchitecture, as demonstrated by histopathological assessment. These findings highlight quercetin’s therapeutic potential in mitigating aluminium-induced neurotoxicity and suggest its utility in the management of neurodegenerative disorders.

The aim of the presented study was to investigate the effects of prebiotic inulin, probiotic VSL#3 (mixture of bacteria from 7 different species and 8 strains) and synbiotic (inulin + VSL#3) supplements applied together with lacosamide (LCM) on post-traumatic epilepsy (PTE). In addition, effectiveness of the relevant treatments on comorbid problems related to learning and memory, anxiety, motor performance and pain threshold that may develop together with seizures due to traumatic brain injury (TBI) and PTE was also examined using behavioral tests. In experiments, adult male Sprague-Dawley rats, divided into 6 groups, were given 30mg/kg LCM or 1000mg/kg inulin together with LCM as prebiotic, VSL#3 mixture containing 10 × 109 CFU/kg bacterial colonies as probiotic and (inulin + VSL#3) as synbiotic for 28 days by oral gavage after mild-TBI was induced by weight-drop method and electroencephalogram electrodes were placed. In order to reveal the tendency to trauma-induced seizures, PTE was modeled by injection of 30 + 15 + 15mg/kg pentylenetetrazole at subconvulsant doses at 30-min intervals on 7th, 14th, 21st and 28th days after TBI and seizures were scored according to Racine scale. Locomotor, cognitive and pain-related behaviors of animals other than seizures were also examined using open field, rotarod, elevated plus maze, radial arm maze, dynamic plantar and thermal plantar tests. It was determined that LCM applied alone was effective in seizure control related to PTE, but probiotic supplement in form of LCM + VSL#3 provided more effective seizure control. In addition, it was determined that probiotic VSL#3 applied together with LCM increased locomotor activity, both VSL#3 and inulin prevented the disorder in forced motor activity caused by LCM, and VSL#3 + inulin applied together with LCM increased mechanical pain threshold. It was concluded that probiotic VSL#3 combined with antiepileptic LCM provides more effective seizure control by strengthening the antiepileptic activity of LCM in management of PTE.

Persisting neurobehavioral consequences of gestational exposure to cadmium and benzo[a]pyrene in rats.

BackgroundAccumulating evidence suggests that selective activation of the M1 and/or M4 muscarinic acetylcholine receptor (mAChR) subtypes may offer a novel therapeutic approach for the treatment of psychotic symptoms and cognitive impairments observed in psychiatric disorders, including schizophrenia. The M1 and M4 receptors are members of the family A G protein-coupled receptors; and include five different subtypes termed M1–M5, which modulate many central nervous system functions, including affective responses and cognition. Unfortunately, until the recent clinical success of CobenfyTM, M1 and/or M4 muscarinic orthosteric agonists ultimately failed in clinical development due to the lack of receptor subtype selectivity resulting in adverse side effects associated with non-selective activation of peripheral M2 and M3 mAChRs.Aims & ObjectivesThis presentation will review recent breakthrough innovations by researchers at the Warren Center for Neuroscience Drug Discovery to identify subtype-selective M1 and M4 mAChR tool compounds and clinical candidates that activate a specific receptor subtype at sites that are less highly conserved than the orthosteric binding site of acetylcholine (ACh) and more topographically distinct, termed allosteric sites. These efforts have led to the discovery of highly, selective M1 and M4 positive allosteric modulators (PAMs), ligands that do not activate the M1 or M4 receptor directly, but markedly potentiate the response of the receptor subtype to ACh.MethodHTS, medicinal chemistry and cheminformatics techniques were utilized to identify and optimize selective M1 and M4 mAChR PAMs, including VU0467319 and VU0467154 respectively; as well as selective M4 mAChR orthosteric antagonists, represented by VU6028418. In vitro cell-based assays were used to generate functional potency, efficacy, and selectivity data. Pharmacokinetic (PK) studies measured compound plasma/brain exposure in preclinical species using LC/MS detection. Compound efficacy was assessed in rodent models of antipsychotic-like activity, cognitive enhancement, and changes in sleep/wake architecture with DSI telemetry devices.ResultsIn vitro studies revealed that VU0467319 is a moderately potent M1 PAM (EC50 = 492 nM, ACh max=71%) with minimal M1 agonism (EC50 > 30 μM), and no activity at other mAChRs up to 30μM. VU0467319 exhibited excellent multispecies PK and produced dose-dependent increases in prefrontal cortical- and hippocampal-mediated learning and memory tasks in rodents, including novel object recognition, contextual fear conditioning and eight arm radial arm maze tasks; as well as increased high frequency gamma power, a correlate of enhanced arousal. In vitro studies demonstrated that VU0467154 is highly potent (EC50=18nM at rM4; ACh max=68%) and selective M4 PAM with no activity at other mAChRs up to 30μM. VU0467154 produced reversal of psychostimulant-induced hyperlocomotion in wildtype mice and rats, but not in M4 KO mice. VU0467154 also reversed impairments in PFC-dependent cognitive tasks, including visual pairwise discrimination. VU0467154 increased non-rapid eye movement sleep, while decreasing rapid eye movement sleep and arousal; all effects were attenuated using the M4 orthosteric antagonist VU6028418. VU0467319 and VU0467154 did not induce any dose-limiting adverse side effects.Discussion & ConclusionsThe pharmacologic characterization of selective M1 and M4 mAChR PAMs has revealed distinct efficacy profiles that may provide innovative treatment strategies for addressing the different affective and cognitive impairments observed in schizophrenia and other neuropsychiatric disorders.

BackgroundOrexin neurons regulate physiological functions, including not only energy homeostasis and wakefulness, but also motivated behaviors. However, it is unknown whether these neurons are involved in reward choice strategy. We recently developed a transgenic rat line that expresses Cre recombinase exclusively in orexin neurons.Aims & ObjectivesHere, we examined the roles of orexin neurons in reward choice behavior under conditions of uncertainty in a gambling test in rats, and analyzed the changes in choice behavior when orexin neurons were manipulated using chemogenetic approaches.MethodWe developed a transgenic rat line that expresses Cre recombinase exclusively in orexin neurons. We used rat gambling test using a radial arm maze to examine decision-making strategies.ResultsIn the gambling test, cell-specific excitatory manipulation of orexin neurons using chemogenetics resulted in risky arm choice. Positive, but not negative, reward prediction error contributed to reward-based risky choice when orexin neurons were activated. These risky behaviors were inhibited by suvorexant, orexin receptor antagonist, treatment. Suvorexant alone also induced conservative choice behaviors.Discussion & ConclusionsOur data suggest that activated orexin neurons affect the processing of reward prediction error and alter strategy in reward-based decision-making.

The present study explores the innovative application of virtual reality (VR) in conducting the Radial Arm Maze (RAM) task, a performance-based test traditionally utilized for assessing spatial memory. This study aimed to develop a gamified version of the RAM implemented in immersive VR and investigate the interaction between the sense of presence, movement features, and performance within the RAM. We developed software supporting a head-mounted display (HMD), addressing prior limitations in the scientific literature concerning user interaction, data collection accuracy, operational flexibility, and immersion level. This study involved a sample of healthy young adults who engaged with the immersive VR version of the RAM, examining the influence of VR experience variables (sense of presence, motion sickness, and usability) on RAM performance. Notably, it also introduced the collection and analysis of movement features within the VR environment to ascertain their impact on performance outcomes and their relationship with VR experience variables. The VR application developed is notable for its user-friendliness, adaptability, and integration capability with physiological monitoring devices, marking a significant advance in utilizing VR for cognitive assessments. Findings from our study underscore the importance of VR experience factors in RAM performance, highlighting how a heightened sense of presence can predict better performance, thereby emphasizing engagement and immersion as crucial for task success in VR settings. Additionally, this study revealed how movement parameters within the VR environment, specifically speed and directness, significantly influence RAM performance, offering new insights into optimizing VR experiences for enhanced task performance.

This method paper details a protocol to test spatial working memory in mice using a semi-automated radial 8-arm maze (RAM). The RAM is a partially enclosed apparatus with 8 horizontal, equally spaced arms radiating from a central hub, from which access to each arm can be controlled individually by servo-controlled motorized doors. Animals start in the central hub and are allowed to explore the maze for a food reward at the end of each arm or selected arm. The RAM task was originally designed for rats, but we have adapted the protocol for mice, for example, by including more habituation steps. In our protocol, all arms are initially baited with sweetened condensed milk, and mice are admitted sequentially into four pseudo-randomly selected arms to collect the rewards ("forced run") before opening all doors together to allow the mice to run freely and find the remaining four rewards ("free run"). A 5 s delay is introduced between the forced and free runs to challenge working memory; an error is recorded if the mouse enters any previously visited arm during the free run. The task is complete when all rewards are recovered. After 6 days of habituation and 9 days of maze training, male C57BL/6 mice regularly achieve ≥ 80% daily success rate score, defined as 4/(4+E), where E is the number of errors. This semi-automated task could, in principle, be combined with in vivo monitoring methods such as electrophysiology, multiple-photon microscopy, or calcium imaging.

Numerous studies have looked into the neurocognitive impact of cannabinoid ligands. The effect of Catha edulis (Vahl) Forssk. ex Endl. (khat) on learning and memory is also researched by few scholars, although the findings are inconsistent. Moreover, the interaction of cannabis with other drugs of abuse is well studied. According to recent reports, the endocannabinoid system modulates the neurobehavioural effects of khat, including motor activities, working memory, and anxiety-like behaviours. However, it is not known whether such modulation is apparent in spatial learning and memory. To this effect, mice were subjected to acute exposure to khat and cannabinoids (agonists and antagonists) and spatial learning and memory were assessed using a battery of tests including the radial arm maze, multiple T-maze and the Morris water maze. Parameters including latency, correct/incorrect decision, reference memory error, and working memory error were determined. The study revealed that acute khat exposure did not have a substantial effect on spatial learning and memory. Method-dependent pro-cognitive effect was observed with single administration of JWH-133 (CB2R agonist) and cannabidiol. By contrast, WIN-55,212-2 (a nonselective CBR agonist) resulted in significant impairment of cognition. Whereas, co-administration of khat with cannabinoid agonists attenuated the effect produced by the agonists regardless of the direction of change(enhanced or reduced cognition). Co-administration with inverse agonists/antagonists, however, had a procognitive effect. Especially, co-administration of khat and the cannabinoid 1 receptor inverse agonist/antagonist AM-251 improved spatial learning and memory. Overall, this research indicates that khat could potentially be able to modify how the endocannabinoid system affects spatial learning and memory.

Integrated network pharmacology, metabolomics, and microbiome studies to reveal the therapeutic effects of Anacyclus pyrethrum in PD-MCI mice.

Methiopropamine (MPA), a novel psychoactive substance (NPS) similar to methamphetamine (METH), warrants investigation into its neurotoxic effects on cognitive function and behaviors due to limited existing research. Therefore, this study aimed to explore the effects of MPA on several behavioral parameters in mice, brain levels of monoamine neurotransmitters, and p-ERK1/2 expression. Mice were randomly divided into four groups (n = 10) which received daily intraperitoneal injections of either saline, 1 or 3 mg/kg of MPA, or 1 mg/kg of METH for 7 days. The novel object recognition test (NORT) revealed a significant decline in recognition memory, particularly evident at a dose of 3 mg/kg of MPA, similar to METH at 1 mg/kg, observed 24 h post-withdrawal. MPA at 3 mg/kg also impaired working and reference memory performance in the 8-arm radial maze (8-ARM) test and exhibited an anxiolytic effect in the open field test (OFT). These cognitive impairments were accompanied by decreased dopaminergic parameters and p-ERK1/2 expression within the prefrontal cortex (PFC). This further suggests that MPA neurotoxicity is targeted at the dopaminergic transmission in the PFC. In conclusion, MPA consumption is associated with memory impairment, which is attributable to dopaminergic deficits and reduced p-ERK1/2 activities in the PFC.

Aging is accompanied by a decline in cognitive functions, including spatial memory, yet significant variability exists in the learning abilities of older individuals. Using a large cohort of aged and young male mice, we employed spatial discrimination testing in an 8-arm radial maze to investigate age-related differences in performance in spatial learning and to categorize individual memory phenotypes within the aged population. Despite a general learning ability across groups, aged mice showed slower acquisition rates compared to young counterparts, highlighting age-related cognitive difficulties in establishing or discriminating spatial representations. By modeling individual learning curves, we classified aged mice into two subgroups-normal learners (NL) and slow learners (SL)-based on learning speed. SL mice demonstrated significantly delayed spatial memory acquisition compared to NL and young mice, suggesting pronounced heterogeneity in cognitive aging. This method provides a robust framework to explore the neurobiological underpinnings of learning deficits and may inform the development of targeted interventions to mitigate age-related memory decline.

Amyloid deposits initiate neuroinflammation by activating astrocytes and microglia in the hippocampus, increasing neuronal vulnerability and loss. Astrocytes, while essential for cerebral function, can contribute to neuronal dysfunction by retracting neuronal synapses, that forms a consequence of neuroinflammation, leading to cognitive deficits in Alzheimer's disease (AD). Upon Amyloid-β (Aβ) deposition, astrocytes become reactive as part of a repair mechanism, however this process can impair neurogenesis resulting in AD progression. The current study hypothesizes that resveratrol (RSV) can address inflammation and promote neural regeneration, mitigating cognitive decline. Our previous research highlights RSV's homeostatic effect through SIRT1 normalization, which is crucial in preventing AD progression. However, its neurogenic potential in AD remains underexplored. In this study, Aβ25-35-induced AD rat model was used to study the anti-inflammatory, neurogenic and cellular homeostatic effect of RSV (30mg/kg) for four weeks. Results showed increased Doublecortin expressing cells, indicating favorable neurogenesis in hippocampus. Immunofluorescence of microglia and astrocytes in the hippocampus revealed that RSV counteracted their activation by reducing the formation of engulfing microglia and elongated astrocytes. Behavioral assessments using the Morris water maze and cued radial arm maze demonstrated significant improvements in spatial and learning memory. These cognitive improvements were supported by increased choline acetyltransferase and SIRT1 levels. These findings suggest that RSV effectively reduces neuroinflammation, promotes neurogenesis in the sub granular zone of the hippocampus, and improves learning and memory in both control and AD conditions via SIRT1. This study highlights RSV's potential as a suitable therapeutic agent for AD.

Background: Memory impairment involves a decline in memory, cognition, behavior and daily functioning. Conventional treatments often fall short due to the complex mechanisms underlying memory loss, diminishing effectiveness over time and having significant side effects. In this context, medicinal herbs have gained attention for their broad therapeutic benefits and lower risk of adverse effects. Among them, Azadirachta indica leaf extract (AILE) is notable for its diverse biological properties, making it a promising candidate for addressing cognitive decline. Objective: The study aimed to evaluate the effects of AILE on spatial working and reference memory in male Wistar rats with ketamine-induced memory impairment. Ketamine, a known NMDA receptor antagonist, was used to induce cognitive deficits, which were assessed using the Radial Arm Maze (RAM). Methods: This experimental study was conducted in the Department of Physiology at Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka, using 30 male Wistar rats (200±50 g body weight) sourced from the university's central animal house. The rats were divided into three groups: Group 1 (G1) normal memory, Group 2 (G2) memory impaired, Group 3 (G3) experimental, respectively. Ethical approval for this research was obtained from the Institutional Review Board (IRB) of BSMMU. Data were analyzed using ANOVA, Bonferroni post hoc tests, and Student's paired t-test with significance set at p≤0.05. Results: In the RAM test, ketamine-treated rats exhibited a significant increase in both working memory errors and reference memory errors (p≤0.001), indicating substantial memory impairment. However, rats treated with AILE showed a significant reduction in both working memory errors and reference memory errors (p≤0.001) compared to the ketamine-only group. These results suggest that AILE effectively mitigates ketamine-induced cognitive deficits, improving both working and reference memory performance in the RAM. Conclusion: AILE demonstrated significant neuroprotective effects against ketamine-induced memory impairment, likely through modulation of NMDA receptor function, reduction of oxidative stress, and inhibition of apoptotic pathways. These findings suggest AILE's potential as a therapeutic agent for cognitive deficits involving NMDA receptor dysfunction.

Status epilepticus causes the most severe condition related to epilepsy in terms of high mortality rate. Although status epilepticus treatment guidelines specify a treatment process based on three-stage monotherapy, effective control cannot yet be achieved in all cases. In the presented study, with electrophysiological and behavioral tests, it was aimed to investigate the effectiveness of the combination of midazolam (MDZ), one of the most commonly used benzodiazepines in the first-line treatment of status epilepticus, with the second-line antiepileptics levetiracetam (LEV), lacosamide (LCM), valproic acid (VPA), and fosphenytoin (fPHT). A status epilepticus model was created with lithium-pilocarpine (5mEq/kg-320mg/kg) in adult male Sprague-Dawley rats with implanted electroencephalography (EEG) electrodes. MDZ (9mg/kg) alone or in dual combinations with antiepileptic drugs (200mg/kg LEV, 50mg/kg LCM, 300mg/kg VPA, 100mg/kg fPHT) was injected i.p. to the experiment groups with status epilepticus. After video-EEG recordings were taken from the rats during and after status, the effects of drug interactions on cognitive and motor behaviors were examined by applying behavioral tests (open field, Rotarod, radial arm maze, and passive avoidance). Compared with the untreated status epilepticus group, it was determined that MDZ alone and the combination of four antiepileptic drugs administered with MDZ significantly reduced the mortality rate, spike frequency, and spike amplitude of epileptic seizures and suppressed epileptic seizures at certain levels (p<0.01). Compared to MDZ monotherapy, it was determined that the mortality rate and spike frequency and amplitude decreased significantly in the MDZ+LCM group (p<0.01), whereas on the other hand, mortality and spike frequency increased in the MDZ+LEV group (p<0.01). No negative effects were observed in learning and memory in all treatment groups, but it was determined that the motor functions of the animals treated with MDZ+fPHT were impaired compared to both the control group without any treatment and the MDZ group (p<0.01). In the status epilepticus model induced by lithium-pilocarpine, the combination of MDZ+LCM was found to be the most effective polytherapy option in reducing seizures and mortality. Additionally, it was observed that LEV, LCM, and VPA administered together with MDZ did not negatively affect both cognitive and motor functions.