Brain Levels Research Articles

Co-administration of coenzyme Q10 and curcumin mitigates cognitive deficits and exerts neuroprotective effects in aluminum chloride-induced Alzheimer's disease in aged mice.

Aluminum chloride (AlCl3), a known neurotoxic and Alzheimerogenic metal disrupts redox homeostasis which plays a pivotal role in pathophysiology of neurodegenerative disorders, particularly cognitive decline. The current study was designed to unveil the long-term neuroprotective outcomes and efficacy of CoQ10 and curcumin low dose (100 mg/kg each) combination in 18-months old geriatric male Balb/c mice subjected to AlCl3-prompted memory derangements (200 mg/kg in water bottles) for 28 days. The neuroprotective properties driven by antioxidant mechanisms were assessed via observing cellular pathology in key-memory related brain regions including the cornuammonis (CA3 and DG) and cortex 2/3 layer. Our outcomes revealed that AlCl3 exposure significantly reduced spatial learning and memory. In contrast, CoQ10 and curcumin combinatorial regime markedly mitigated cognitive deficits Vs. individual high-dose in AlCl3-treated animals as demonstrated by their improved performance in neurobehavioral tests such as the Y-maze, novel object recognition, passive avoidance and Morris-water maze test. Additionally, CoQ10 and curcumin co-administration restored redox balance by significantly reducing the levels of oxidative stressor (MDA) and increasing the anti-oxidant capacity (SOD,GPx). AchE is an enzyme involved in acetylcholine breakdown which negatively impacts acetylcholine levels and memory function. AlCl3 exposure elevated AchE levels in mice brains vs. treatment. This neurochemical alteration was notably reversed in the dual-treatment group. Furthermore, CoQ10 and curcumin ameliorated AlCl3-induced neurotoxicity by preserving neuronal cytoarchitecture in both cortical and hippocampal regions. In conclusion, CoQ10 and curcumin combination might attenuate memory loss induced by AlCl3-intoxication via restoring aberrant AchE activity, enhanced anti-oxidant defenses and salvaging the deleterious neuronal damage.

Cerebral blood flow following successful living kidney transplantation: VINTAGE study

Abstract Background Chronic kidney disease (CKD) is a significant risk factor for cerebrovascular disease. However, there is limited research on how successful living donor kidney transplantation (LDKT) affects cerebral blood flow (CBF). This study aims to comprehensively investigate how LDKT influences CBF across various brain levels and regions. Methods Data from 53 recipients between 2016 and 2020 were obtained from the VINTAGE study conducted at our hospital. CBF was measured by level and region using single-photon emission computed tomography (SPECT), according to the Talairach brain atlas. The primary endpoint was the mean difference in CBF before and 1-year post-LDKT. Subgroup analysis using traditional risk factors assessed the heterogeneity of the effect on CBF in the frontal lobe region. Results LDKT improved blood flow in the anterior cerebral artery and middle cerebral artery but had less impact on the posterior cerebral artery. The most consistent improvements were observed in the frontal lobe region (left frontal lobe: -0.12 [95% CI -0.18 to -0.05, P < 0.001]; right frontal lobe: -0.13 [95% CI -0.21 to -0.05, P = 0.001]. Subgroup analysis showed a consistent effect of LDKT on frontal lobe CBF improvement, with no qualitative interaction observed. Conclusions LDKT contributes to the normalization of CBF, with improvement in anterior circulation and frontal lobe blood flow. To clarify the clinical significance of KT's CBF-improving effect, future studies should investigate the relationship between specific cognitive impairments (e.g. short-term memory, visuospatial ability, executive function) and CBF in each perfusion region.

How does teaching experience impact brain processes underlying the theory of mind? Study on primary school educators

ABSTRACT Despite its importance for daily social interactions, few studies have explored interindividual differences in the Theory of Mind (ToM) abilities of healthy adults. We used Children’s False-Attribution (CFA), Children’s False-Beliefs (CFB), and Belief-Desire Reasoning tasks, along with fMRI-based assessments, in a comparative analysis of ToM among primary school teachers (PST; n = 27), skilled in social interactions with children, and matched controls (MC; n = 24), who lacked such experience. PST demonstrated slower reaction times than MC in Adult and Child false-belief stories of CFB. However, no other behavioral differences between the groups and between-group differences were observed at the brain level. Both groups presented similar valence ratings for stories in the CFA. Notably, fMRI analysis revealed a group-by-condition interaction effect in the right lateral orbitofrontal cortex (OFC). In PSTs, OFC activation decreased during negative false-attribution stories regarding children compared to stories related to adults, whereas MC demonstrated an opposite activation pattern. Between-group differences in right lateral OFC activity possibly signify a neural efficiency effect secondary to frequent social interactions of PSTs, unlike the MCs, with children in the working environment. These results underscore the significance of everyday social experiences in the functional plasticity of ToM networks.

Brain physiology during photoperiod-related caste determination in the primitively eusocial wasp Polistes jokahamae

Adult females of the primitively eusocial wasp Polistes jokahamae can change caste in response to photoperiod length. Short-day photoperiod females store more lipids but have less developed ovaries for hibernation (becoming gynes), whereas long-day photoperiod females have fewer lipid stores but more developed ovaries under queenless conditions (becoming egg-laying workers). To explore the physiological mechanisms underlying photoperiod-related caste determination, analyses of gene expression levels in the brains of short- or long-day females were performed. Results suggest that short-day females may exhibit higher expression of genes involved in tryptophan metabolism, insulin signaling, and nutrition, including the digestion of sugars and lipids, and production of royal jelly proteins. Oral administration of tryptophan resulted in a positive correlation between tryptophan levels in the brain and lipid stores in the abdomen, suggesting that tryptophan promotes lipid storage in gynes. Long-day females showed enhanced expression of genes involved in tyramine/dopamine syntheses, epidermal growth factor receptor, insulin degradation, and oogenesis. In these females, tyramine administration increased the brain levels of tyramine and dopamine, and activated initial ovarian development under queenless conditions. Together, these results indicate that photoperiod-related caste determination in P. jokahamae may involve gene expression pathways similar to those involved in caste determination during the preimaginal stage in other Polistes species.

NF-κB, neutrophil extracellular traps, and microglial in mice with <em>Streptococcus suis</em> serotype 2 meningitis infection

BACKGROUND Streptococcus suis is the most frequent etiology of zoonotic bacterial meningitis, potentially initiating an outbreak. Acute bacterial meningitis caused by S. suis has various manifestations, often accompanied by sepsis with multiple organ involvement. This study aimed to evaluate the pattern of S. suis outgrowth in the brain, which is associated with nuclear factor-κappaB (NF-κB) activation, neutrophil extracellular traps (NETs) release (NETosis), and microglial activation as three crucial pathological mechanisms of bacterial meningitis. METHODS This study used 64 female BALB/c mice aged 6 weeks and weighed 18−20 g, grouped into infected and non-infected as the control group. Both groups were administered 1 ml of S. suis serotype 2 suspension (1 × 107 colony forming-unit/ml) and normal saline intraperitoneally. The bacterial colony count of S. suis was evaluated, along with NF-κB and NET levels in blood and brain, as well as meningeal inflammation and microglial activation in the brain at Days 1, 3, 5, and 7 post-infection. RESULTS The invasion of S. suis into the brain slightly induced NF-κB activation, leading to a burst of inflammatory responses, neutrophil infiltration with NET releases, and microglia activation that co-occurred, showing their peaks on Days 3 and 5 after onset. CONCLUSIONS The S. suis invasion into the mice’s brain increased NF-κB activation, NETosis, and microglia activation during S. suis meningitis infection.

Group-common and individual-specific effects of structure–function coupling in human brain networks with graph neural networks

Abstract The human cerebral cortex is organized into functionally segregated but synchronized regions bridged by the structural connectivity of white matter pathways. While structure–function coupling has been implicated in cognitive development and neuropsychiatric disorders, it remains unclear to what extent the structure–function coupling reflects a group-common characteristic or varies across individuals, at both the global and regional brain levels. By leveraging two independent, high-quality datasets, we found that the graph neural network accurately predicted unseen individuals’ functional connectivity from structural connectivity, reflecting a strong structure–function coupling. This coupling was primarily driven by network topology and was substantially stronger than that of the correlation approaches. Moreover, we observed that structure–function coupling was dominated by group-common effects, with subtle yet significant individual-specific effects. The regional group and individual effects of coupling were hierarchically organized across the cortex along a sensorimotor-association axis, with lower group and higher individual effects in association cortices. These findings emphasize the importance of considering both group and individual effects in understanding cortical structure–function coupling, suggesting insights into interpreting individual differences of the coupling and informing connectivity-guided therapeutics.

Elevated levels of peripheral and central nervous system immune markers reflect innate immune dysregulation in autism spectrum disorder

BackgroundEvidence suggests dysregulated immune functions in the pathophysiology of Autism spectrum disorder (ASD), although specific immune mechanisms are yet to be identified. MethodsWe assessed circulating levels of 25 immune/neuroinflammatory markers in a large ASD sample (n = 151) and matched controls (n = 72) using linear models. In addition, we performed global brain transcriptomics analyses of relevant immune-related genes. We also assessed the expression and function of factors and pathway elements of the inflammasome system in peripheral blood mononuclear cells (PBMC) isolated from ASD and controls using in vitro methods. ResultsWe found higher circulating levels of IL-18 and adhesion factors (ICAM-1, MADCAM1) in individuals with ASD relative to controls. Consistent with this, brain levels of ICAM1 mRNA were also higher in ASD compared to controls. Furthermore, we found higher expression/activity of Caspase-1 and the inflammasome sensor NLRP3 in PBMCs in ASD, both at baseline and following inflammatory challenge. This corresponded with higher levels of secreted IL-18, IL-1β, and IL-8, as well as increased expression of adhesion factors following inflammasome activation in ASD PBMC cultures. Inhibition of the NLRP3-inflammasome rescued the observed immune phenotype in ASD in vitro. ConclusionOur results suggest a role for inflammasome dysregulation in ASD pathophysiology.

Chemokine associations with blood cerebrospinal fluid barrier permeability and delirium

Delirium is a highly prevalent neuropsychiatric syndrome characterised by acute and fluctuating impairments in attention and cognition. Mechanisms driving delirium are poorly understood but it has been suggested that blood cytokines and chemokines cross the blood brain barrier during delirium, directly impairing brain function. It is not known whether these molecules reach higher brain levels when the blood cerebrospinal fluid barrier (BCSFB) is impaired. Here, in human hip-fracture patients, we tested the influence of BCSFB integrity on CSF levels of chemokines and assessed their association with delirium. CSF levels of IP-10, eotaxin, eotaxin 3 and TARC showed weak to moderate correlations with BCSFB permeability, as measured by the Qalbumin ratio, while MCP1, IL-8, MIP1α and MIP1β showed no significant correlation. Chemokines were not associated with delirium in univariate analysis or when stratified on dementia status, but exploratory analyses showed that elevated Eotaxin (CCL11) and MIP1α (CCL3) were associated with prevalent delirium. Modelling acute systemic inflammation, we used bacterial LPS (250 μg/kg) or sterile laparotomy surgery in mice to demonstrate de novo synthesis of chemokines at the choroid plexus (CP) and microvasculature. Gene expression data showed CP-enriched expression of Il1b, Tnfa, Cxcl1 and Ccl3 in both models and immunohistochemistry showed cytokine and chemokine synthesis in CP stromal (IL-1β, CCL2/MCP1) or epithelial cells (CXCL10/IP-10) cells and at the microvasculature. Larger studies are required to confirm these human findings on chemokine associations with BCSFB permeability and prevalent delirium. Preclinical studies are warranted to determine whether chemokines might play a role in the pathophysiology of delirium.

Therapeutic impact of a benzofuran derivative on Aluminium chloride-induced Alzheimer's disease-like neurotoxicity in rats via modulating apoptotic and Insulin 1 genes

Neurodegenerative disorders such as Alzheimer’s disease (AD) are age-related and are fatal in advanced cases. There is a limited efficacy of drugs used for the management of these diseases. Herein, the neurotherapeutic efficacy of a benzofuran-derivative-7 (BF-7) was investigated. Aluminum chloride (AlCl3) was employed to induce AD-like brain toxicity in rats. The rats were divided into four groups: Negative control, AlCl3-induced AD rats (100 mg/kg body weight, orally), AlCl3-AD induced rats treated with BF-7 (10 mg/kg body weight, orally), AlCl3-AD-induced rats treated with the standard drug “Donepezil” (10 mg/kg body weight, orally). The behavioral performance was tested using a beam-balance test. Brain and serum acetylcholinesterase (AChE) activities and the brain levels of norepinephrine, dopamine (DA), and serotonin (5-HT) were measured. The genetic expression of Bcl-2, Bax, caspase-3, and insulin 1 were assayed. The histopathological imaging and the immunohistochemical evaluation of Glial Fibrillary Acidic Protein (GFAP) were investigated in the cerebral cortex. Treatment of AD-rats with BF-7 mitigated AlCl3-induced neurotoxicity by improving motor functions, counteracting apoptosis, and exerting cholinergic functions. In addition, the genetic expression of Insulin 1 was upregulated significantly in AD-induced rats treated with BF-7. This compound could be used as a promising candidate for neurotherapeutic drug discovery against AD or any other toxic brain disorders.

Gastrodin reduces Aβ brain levels in an Alzheimer's disease mouse model by inhibiting P-glycoprotein ubiquitination

BackgroundStudies have demonstrated the potential of gastrodin in the treatment of Alzheimer's disease (AD), however, its mechanism of action remains elusive. Currently, the Amyloid-β (Aβ) cascade hypothesis continues to be the prevailing theory regarding AD etiology. The ubiquitination of P-glycoprotein (P-gp) at the blood-brain barrier (BBB) contributes to the accumulation of Aβ in the brain during AD. PurposeTo investigate the mechanism of gastrodin intervention in AD. MethodsThe molecular docking, molecular dynamics simulations, and microscale thermophoresis (MST) were employed to identify the action target of gastrodin. The western blot (WB) was performed to detect the protein expression level, the ubiquitination level of P-gp was determined using co-immunoprecipitation (CO-IP) assay. P-gp transport activity was detected using an NBD-CSA fluorescence assay. Trans-Epithelial Electrical Resistance (TEER) was used to detect cell resistance. Fluorescein-labeled dextran experiments were performed to determine the individual cell permeability. The immunofluorescence (IF) was employed to detect Aβ deposition, the Morris Water Maze test was used to assess behavioral changes in APP/PS1 mice and the levels of Aβ40 and Aβ42 expression were quantified using enzyme-linked immunosorbent assay. ResultsThe FBXO15 was the target of gastrodin-mediated inhibition of P-gp ubiquitination. Gastrodin increased the P-gp expression, cell resistance, and P-gp transport activity of BEND.3 cells upon treatment with Aβ40 through mechanisms involving the reduction of FBXO15 and P-gp binding and the inhibition of P-gp ubiquitination. And gastrodin could effectively improve memory function and increase number of neurons in APP/PS1 mice, reduce the accumulation of Aβ40 and Aβ42, and enhance P-gp expression in a dose-dependent manner. ConclusionAβ40 induces the ubiquitination and proteasomal degradation of BBB P-gp, however, gastrodin inhibits the ubiquitination of P-gp by binding to FBXO15, thereby increasing P-gp protein expression and enhancing its transport function.

Anti-oxidative, anti-AChE and neuroprotective effect of hydro-alcoholic extract of Sansevieria cylindrica leaves and Plumeria obtusa seed pods in Wistar rats

The rising incidence of neurodegenerative diseases represents a significant global health concern. The prognosis for such diseases is often unfavorable, and patients require supportive treatment to manage their condition effectively. The present study investigated the anti-oxidative, anti-acetylcholinesterase (anti-AChE) effect of hydro-alcoholic extract of Sansevieria cylindrica leaves and Plumeria obtusa seed pods in Wistar rats. Plants were authenticated followed by pharmacognostic evaluation. The antioxidant and anti-AChE activities were evaluated by scopolamine-induced oxidative stress and acetylcholine level alterations in Wistar rats. On the 0th, 7th, and 14th days during study period behavioral parameters were evaluated, including open field test, new object recognition test, and Morris water maze task. Different antioxidant enzymes were also evaluated in brain homogenate post 7 days treatment. AChE levels in rats’ brains were assessed post 14 days of treatment. Finally, histopathology of brains samples was performed. Correspondingly, in-vitro and in-silico studies were done to support the study findings.ResultsSansevieria cylindrica and Plumeria obtusa exhibited strong antioxidant and anti-AChE activities due to secondary metabolites content. Rearing and, line crossing by rats showed substantial alteration (p < 0.05) by combination of both plants at high dose. The escape latency found to be reduced significantly (p < 0.05) by Sansevieria cylindrica individually and in combination with Plumeria obtusa at all doses. The lipid peroxidation level in brain was decreased significantly than standard at 1:1 combination of plants extracts. The levels of GSH, CAT and SOD were attenuated significantly by different concentration of extracts (p < 0.05 and p < 0.001, respectively). The reduction in AChE levels was around 34.62% and 31.10% in male and female rats, respectively in combination of both plants at high dose compared to disease control group. It was further evident in brain histopathology. All in-vivo results were supported by in-vitro free radicals scavenging effect and in-silico study.ConclusionThe hydro-alcoholic extracts from Plumeria obtusa seed pods and Sansevieria cylindrica leaves showed a strong natural antioxidant, and acetylcholinesterase inhibitions which can aid in the treatment of neurodegerative diseases including Alzheimer.

Sex-specific perturbations of neuronal development caused by mutations in the autism risk gene DDX3X.

DDX3X is an X-linked RNA helicases that escapes X chromosome inactivation and is expressed at higher levels in female brains. Mutations in DDX3X are associated with intellectual disability (ID) and autism spectrum disorder (ASD) and are predominantly identified in females. Using cellular and mouse models, we show that Ddx3x mediates sexual dimorphisms in brain development at a molecular, cellular, and behavioral level. During cortical neuronal development, Ddx3x sustains a female-biased signature of enhanced ribosomal biogenesis and mRNA translation. Female neurons display higher levels of ribosomal proteins and larger nucleoli, and these sex dimorphisms are obliterated by Ddx3x loss. Ddx3x regulates dendritic outgrowth in a sex- and dose-dependent manner in both female and male neurons, and dendritic spine development only in female neurons. Further, ablating Ddx3x conditionally in forebrain neurons is sufficient to yield sex-specific changes in developmental outcomes and motor function. Together, these findings pose Ddx3x as a mediator of sexual differentiation during neurodevelopment and open new avenues to understand sex differences in health and disease.

Brain metabolic profiling of schizophrenia: a path towards a better understanding of the neuropathogenesis of psychosis.

Schizophrenia (SCZ) is a complex psychotic syndrome whose pathogenesis involves countless protagonists, none of which, to date, can fully explain how this disorder develops. In this narrative review, an overview of the biochemical impairment is offered according to several perspectives. Indeed, the metabolic framework behind SCZ dopaminergic hypotheses, glutamate - gamma-amynobutyric acid dysregulation, norepinephrine and serotonin, calcium channel dysfunction is addressed together with the energetic impairment, involving glucose and lipids in SCZ etiopathogenesis, in order to highlight the multilevel pathways affected in this neuropsychiatric disorder. Furthermore, neuroinflammation is analyzed, by virtue of its important role, widely investigated in recent years, in neurodegeneration. Tracing the neurotransmitter activity at the brain level by assessing the metabolic network behind the abovementioned molecules puts into light as unavoidable the need for future studies to adopt an integrate approach to address SCZ pathological and clinical picture. The combination of all these factors, essential in acquiring an overview on the complexity of SCZ pathophysiology represents a crucial step in the development of a more targeted management of SCZ patients.

Limosilactobacillus reuteri ameliorates maternal separation stress in newborn mice and alters subsequent adult behaviour.

Maternal separation (MS) in mice results in behavioral deficits and gut microbiota dysbiosis that all persist into adulthood. Limosilactobacillus reuteri DSM 17938 modulates gut microbiota, alters systemic metabolites, and facilitates immune regulation. To assess the effect of DSM 17938 on biochemical and behavioural stress-associated changes, newborn mice were exposed to unpredictable MS (MSU) daily from day 7 to day 20 of life, with intragastric administration of DSM 17938 or PBS as control. Body weight, brain levels of cholecystokinin (CCK), glial fibrillary acidic protein (GFAP), corticosterone, and stool microbiota were assessed at day 21. Behaviour tests including Y-maze (YMT), Tail Suspension (TST), and Open Field (OFT) were evaluated in adult mice. MSU resulted in a decrease in early postnatal growth, which improved with DSM 17938. Reduced CCK and increased corticosterone brain levels due to MSU were reversed by DSM 17938. GFAP levels increased with MSU, indicating that the decreased brain CCK was likely secondary to neuronal damage. DSM 17938 treated offspring demonstrated better cognitive function and less anxious behaviour in adult behaviour tests. DSM 17398 corrected stress related gut microbial dysbiosis. In conclusion, early life modulation of gut microbiota by DSM 17938 had beneficial effects on stress-associated physical and biochemical changes caused by MS in neonates and on subsequent adult behaviour.

Andrographolide prevents acute hyperammonemia-induced motor dysfunction in rats. Evidences for its mechanism of action

BackgroundBeside the cirrhotic one, non-cirrhotic hyperammonemia (HA) can also cause neuro-degeneration and establishment of 'Hyperammonemic Encephalopathy', ultimately leading to the coma stage. We have done pilot study on aqueous extract of Andrographis paniculata herb against HA induced oxidative stress in rats. Andrographolide (AGP) is the major component isolated from of A paniculata and reported to have various biological activities. PurposePresent study aimed to explore the therapeutic potentiality of AGP on rat model of non cirrhotic acute HA. MethodsRotarod and traction test was performed to access the motor function. Brain and serum ammonia level were assayed to confirm occurrence of HA. Malondialdehyde (MDA), reduced glutathione (GSH), protein carbonylation levels and activity of superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx), glutathione reductase (GR) were studied as oxidative stress markers. Expression of two NMDAR subunits, NR2A, 2B, nNOS and level of nitrite (NO2), cGMP in cerebral cortex and cerebellum were assayed to study the NMDAR-nNOS-cGMP cascade. ResultAcute HA caused motor dysfunction in rats. MDA level and protein carbonylation, increased significantly while SOD, catalase, GPx and GR got suppressed and NMDAR-nNOS-cGMP cascade activated in cerebral cortex and cerebellum. All these suggested establishment of hyperammonemic encephalopathy. AGP treatment restored the motor function, lowered the ammonia level, suppressed the oxidative stress markers, NR2A, 2B, nNOS over-expression and elevated NO2, cGMP levels in cortex and cerebellum. ConclusionOur study showed AGP could be a potential natural anti-hyperammonemic drug as it has prevented the development of acute hyperammonemic encaphalopathy in rats by suppressing NMDAR-nNOS-cGMP cascade and oxidative stress in cerebral cortex and cerebellum.

Effect of JM-20 on Age-Related Cognitive Impairment in Mice.

The decline in cognitive function associated with aging significantly impacts the well-being of elderly individuals and their families. This decline is a major recognized risk factor for neurodegenerative diseases, notably Alzheimer's disease. Animal models of aging provide a platform for evaluating drugs concerning aspects like memory and oxidative stress. JM-20 has demonstrated protective effects on short-term memory acquisition and consolidation, along with antioxidant properties and modulation of Acetylcholinesterase activity. This study assesses the potential protective JM-20 against cognitive decline and age-related memory loss. For the study, aged mice exhibiting aging-associated damage were initially selected. Experimental groups were then formed, and the effect of 8mg/kg of JM-20 was evaluated for 40 days on aging-related behavior, such as spatial memory, novelty recognition memory, ambulatory activity, and anxiety. Subsequently, animals were sacrificed, and the hippocampal region was extracted for redox studies and to assess acetylcholinesterase activity. Results indicated that JM-20 at 8mg/kg reversed damage to spatial working and reference memory, exhibiting performance comparable to untreated young adult animals. Furthermore, JM-20 preserved the enzymatic activity of superoxide dismutase, catalase, and total sulfhydryl levels in age-related cognitive impairment in mice, indicating a potent protective effect against oxidative events at the brain level. However, only young, healthy animals showed decreased acetylcholinesterase enzyme activity. These findings provide preclinical pharmacological evidence supporting the neuroprotective activity of JM-20, positioning it as a promising therapeutic candidate for treating memory disorders associated with aging.

Preclinical metabolism and disposition of [14C]GFH009, a novel selective CDK9 inhibitor

GFH009 is a potent and highly selective cyclin-dependent kinase 9 (CDK9) inhibitor currently under phase II clinical trials. In this study, we investigated the metabolism and disposition of GFH009 in Sprague-Dawley (SD) rats, as well as in vitro metabolism of CD-1 mouse, SD rat, beagle dog, cynomolgus monkey, and human. A radiolabelled study indicated that [14C]GFH009 was quickly and widely distributed throughout the body, but presented low levels in brain, testis, and epididymis after a single intravenous dose of 6 mg (100 µCi)/kg to SD rats. GFH009 undergoes systemic metabolic changes, primarily through O-demethylation, oxidation to carboxylic acid and N-dealkylation, cleavage off the methoxyisopropyl moiety being a minor pathway. These metabolic pathways were found to be mainly consistent both in vitro and in vivo. In SD rats, GFH009 was rapidly and completely eliminated, with faeces serving as the major excretion pathway and urine serving as the minor one. Besides, the major clearance pathway for GFH009 was excretion and the minor one was metabolism. GFH009 exhibits favourable drug metabolism and pharmacokinetics (DMPK) properties, which provides valuable insights into the disposition of GFH009 and can be used to guide future clinical studies.

DDDR-14. IMPROVED ANTI-GLIOBLASTOMA EFFICACY BY BRG399, A NOVEL ORAL MICROTUBULE BINDING AGENT

Abstract Glioblastoma (GBM) is a highly malignant form of brain tumor with a poor prognosis despite advances in treatments. Therapeutic resistance/relapse remains a significant challenge necessitating the development of novel approaches. BRG399 is a brain permeant, non P-gp substrate, microtubule-targeting agent that displays anti-cancer activity in vitro and in vivo. BRG399 was well tolerated in rat and dog toxicology studies. Anti-cancer activity (IC50) of BRG399 in in vitro models of GBM was assessed to have potency ranging from 42nm to 89nM in the 3 cell lines tested. Here, in vivo tumor activity of BRG399 was assessed in Sprague Dawley rats implanted with 3x105 C6 rat glioma cells into the right entorhinal cortex/subiculum region. 14 days post successful implantation tumors were assessed by MRI, rats were randomized to receive vehicle (2% Labrasol) or BRG399 (5, 10, 20 mg/kg groups) twice daily by oral gavage for 16 days, and MRIs were performed on survivors on days 31 and 45 post implantation. BRG399 administration resulted in a marked improvement in survival (from 7.7 ± 2.6 days to 25.5 ± 15.6 days, log-rank test, p:0.007) compared to controls. Six animals receiving BRG399 remained alive beyond day 60 of post-treatment initiation. To assess biodistribution of BRG399 in the brain, rats were implanted with C6 cells and tumor establishment assessed 12 days later by MRI. Animals were treated with vehicle or BRG399 (10, 20, 25 mg/kg) and plasma and brains were collected for bioanalytical assessment. Tissue single cell biodistribution was performed using TIMSTOF FLEX spatial OMICS. These results demonstrate a therapeutic effect of BRG399 on intracranial glioma-bearing rats with survival of the BRG399 treatment groups of strong statistical significance over the control group. Analysis of drug levels in plasma and brain will serve as guidance for dose and scheduling decisions for further therapeutic development.

Supra-Prophylactic Doses of Enoxaparin Reduces Fibrin Deposition Without Exacerbation of Intracerebral Hemorrhage in a Rat Model of Penetrating Traumatic Brain Injury.

Deep vein thrombosis and pulmonary embolism prophylaxis is an important part of trauma care. Despite an increased risk of thrombotic complications, the use of venous thrombosis chemoprophylaxis in penetrating traumatic brain injury (pTBI) patients is met with reluctance from neurosurgeons because of concern for the exacerbation of intracerebral hemorrhage. The objective of this study was to provide initial pre-clinical evidence of the effects of Lovenox (LVX) administration following pTBI with significant intracerebral hemorrhage. Sprague-Dawley rats received a penetrating ballistic-like brain injury. Animals were randomly divided into two groups following injury: LVX (25 mg/kg) or vehicle (VEH, saline). LVX or vehicle was administered subcutaneously beginning 24 h after the injury and continued daily for 7 days post-injury. A neurological assessment was performed daily and magnetic resonance imaging (MRI) was performed at baseline, 1, 2, 3, and 7 days post-injury. Following the final MRI, brains were isolated and prepared for histological analysis. Thromboelastography demonstrated dramatic anticoagulation effects which were confirmed by significant increases in partial thromboplastin time (p < 0.001). Daily neurological assessment revealed no worsening of functional deficits following LVX treatment. MRI analysis demonstrated no differences in cerebral edema or intracranial hemorrhage volumes between treatment groups at any tested post-injury time points. However, LVX elicited a significant reduction in fibrin deposition in the ipsilateral striatum and lesion site at 7 days post-injury (p < 0.05). Serum levels of beta-amyloid were decreased at 7 days following LVX treatment (p < 0.05) which may indicate neuroprotective effects but was not correlated to brain levels. The results presented indicate that administration of LVX at a dose capable of inducing anticoagulation is safe in a rodent model of pTBI without exacerbation of intracerebral hemorrhage within the first 7 days of injury.

"Transcriptional Regulation of human NMNAT2: Insights from 3D Genome Sequencing and Bioinformatics".

Nicotinamide mononucleotide adenylyl transferases 2 (NMNAT2) is a crucial nicotinamide adenine dinucleotide (NAD)-synthesizing enzyme essential for neuronal health. In the Religious Orders Study/Memory and Aging Project (ROSMAP), human brain levels of NMNAT2 mRNA positively correlated with cognitive capabilities in older adults. NMNAT2 mRNA abundance is significantly reduced following various insults or proteinopathies. To elucidate the transcriptional regulation of NMNAT2, we employed circular chromosome conformation capture followed by high-throughput sequencing (4C-seq) to identify potential NMNAT2 enhancer and silencer regions by determining genomic regions interacting with the NMNAT2 promoter in human SH-SY5Y cells. We discovered distinct NMNAT2 promoter interactomes in undifferentiated versus neuron-like SH-SY5Y cells. Utilizing bioinformatics analyses, we identified putative transcriptional factors and NMNAT2-associated genes. Notably, the mRNA levels of many of these genes showed a significant correlation with NMNAT2 mRNA levels in ∼400 single-nuclei RNA-seq datasets from ROSMAP. Additionally, using CRISPR-Cas9 strategies, we confirmed the requirement of two specific genomic regions within the interactomes and four transcription factors in regulating NMNAT2 transcription. In summary, our study identifies genomic loci containing NMNAT2 regulatory elements and predicts associated genes and transcription factors through computational analyses.