Acute Neurotoxicity Research Articles

Preventing acute neurotoxicity of CNS therapeutic oligonucleotides with the addition of Ca2+ and Mg2+ in the formulation

Oligonucleotide therapeutics (ASOs and siRNAs) have been explored for modulation of gene expression in the central nervous system (CNS), with several drugs approved and many in clinical evaluation. Administration of highly concentrated oligonucleotides to the CNS can induce acute neurotoxicity. We demonstrate that delivery of concentrated oligonucleotides to the CSF in awake mice induces acute toxicity, observable within seconds of injection. Electroencephalography (EEG) and electromyography (EMG) in awake mice demonstrated seizures. Using ion chromatography, we show that siRNAs can tightly bind Ca2+ and Mg2+ up to molar equivalents of the phosphodiester (PO)/phosphorothioate (PS) bonds independently of the structure or phosphorothioate content. Optimization of the formulation by adding high concentrations (above biological levels) of divalent cations (Ca2+ alone, Mg2+ alone, or Ca2+ and Mg2+) prevents seizures with no impact on the distribution or efficacy of the oligonucleotide. The data here establishes the importance of adding Ca2+ and Mg2+ to the formulation for the safety of CNS administration of therapeutic oligonucleotides.

Comparative assessment of acute neurotoxicity of real-world ultra-fine black carbon emitted from residential solid fuel combustion

Incomplete combustion of residential solid fuel is one of the main anthropogenic sources for black carbon (BC). Fresh BC, mainly enriched in ultra-fine fraction of particles, can directly cross blood-brain barrier and are reported to be associated with neurodegenerative diseases. Because of the difficulties in collection and purification of BC from ambient particles, there are still significant knowledge gaps in understanding neurotoxicity caused by real-world BC. The purpose of this study is to compare the neurotoxic effects caused by BCs emitted from combustion of six residential solid fuels, and try to reveal associated biological mechanisms in SH-SY5Y cells. Two straw BC (Wheat-BC and Corn-BC) showed highest neurotoxic effects followed by wood BC (Pine-BC and Aspen-BC) and coal BC (Xvzhou and Longkou Coal), as indicated by viability, lactic dehydrogenase, malondialdehyde, adenosine triphosphate and acetylcholine levels. Coal BC caused nearly no toxicity in human neuroblastoma (SH-SY5Y) cells within highest dose of 200 μg/mL. RNA sequence and bioinformatics analysis were applied to effectively identify differential genes and signaling pathways. Based on Gene Ontology (GO) annotation, Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment, Protein Protein Interaction network (PPI network) construction, we found biomass BC affected mitochondrial function, interfered with cellular metabolic processes, disturbed redox homeostasis, and finally resulted in cellular damages. Coal-BC mainly caused cytokine/chemokine related inflammatory responses. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blotting methods were further applied to find out related signaling pathways. Biomass BC activated IL6R/JAK3/STAT3 and JAK3/STAT6 pathways leading to oxidative stress and inflammatory responses. Coal BC activated JAK3/STAT3 pathway leading to chemokine related responses. This study revealed the heterogeneity in neurotoxicity of BCs from different combustion sources and provided important data for health risk assessment. BC-related neurotoxicity should be considered when making air pollution emission control strategies, with residential biomass receiving more policy attention.

Integrating “fast” and “slow” systems therapeutics for physiological renormalization in neurodegenerative disorders

In our previous paper, we provided evidence that simultaneously activating multiple cannabinergic pathways, CB1 and CB2, a systems therapeutic approach, was harnessed to provide “fast” protection from neurodegenerative diseases by renormalizing the physiology of the retina. We had used an animal model of glaucoma, using glutamate-induced neurodegeneration of the retina. Other forms of chronic or acute neurotoxicity throughout the CNS may likewise benefit from a systems approach to cannabinergic mediated rescue of neural tissue. However, from a systems biology perspective, cannabinergic pathways act in many ways, not fully explained in our previous paper, and must act in synergy with many other pathways, including the molecules (e.g. HSP) released by adjacent stem cells, to optimally yield neuroprotection. In addition to the “fast” pathways previously described, we’ll address “slow” acting, lifestyle and nutritional pathways too. Building on our previous paper, we highlight some of the important “fast and “slow” neuroprotective pathways that should be activated together through a systems therapeutic approach.

Dorsal root ganglion inflammation by oxaliplatin toxicity: DPEP1 as possible target for peripheral neuropathy prevention

BackgroundPeripheral neuropathy (PN) constitutes a dose-limiting side effect of oxaliplatin chemotherapy that often compromises the efficacy of antineoplastic treatments. Sensory neurons damage in dorsal root ganglia (DRG) are the cellular substrate of PN complex molecular origin. Dehydropeptidase-1 (DPEP1) inhibitors have shown to avoid platin-induced nephrotoxicity without compromising its anticancer efficiency. The objective of this study was to describe DPEP1 expression in rat DRG in health and in early stages of oxaliplatin toxicity. To this end, we produced and characterized anti-DPEP1 polyclonal antibodies and used them to define the expression, and cellular and subcellular localization of DPEP1 by immunohistochemical confocal microscopy studies in healthy controls and short term (six days) oxaliplatin treated rats.ResultsDPEP1 is expressed mostly in neurons and in glia, and to a lesser extent in endothelial cells. Rats undergoing oxaliplatin treatment developed allodynia. TNF-\U0001d6fc expression in DRG revealed a pattern of focal and at different intensity levels of neural cell inflammatory damage, accompanied by slight variations in DPEP1 expression in endothelial cells and in nuclei of neurons.ConclusionsDPEP1 is expressed in neurons, glia and endothelial cells of DRG. Oxaliplatin caused allodynia in rats and increased TNF-α expression in DRG neurons. The expression of DPEP1 in neurons and other cells of DRG suggest this protein as a novel strategic molecular target in the prevention of oxaliplatin-induced acute neurotoxicity.

A single exposure to brivaracetam or perampanel does not cause cell death in neonatal rats.

Exposure to a range of anti-seizure medications (ASMs) during early brain development adversely impacts neurodevelopmental outcomes in both animal models and in clinical studies. Many ASMs, including phenobarbital, phenytoin, valproate (VPA), and benzodiazepines, are associated with acute neurotoxicity (cell death), impaired synaptic development, and long-term behavioral changes following gestational or neonatal exposure in animals. This is mirrored in clinical studies which show lasting neurodevelopmental deficits following early-life or gestational exposure to these drugs. Brivaracetam (BRV) and perampanel (PER) are two newer generation anti-seizure medications and are of interest based on their mechanisms of action (SV2A modulator, AMPA antagonist, respectively), as other drugs with these mechanisms of action do not trigger acute neurotoxicity. Both BRV and PER show anti-seizure efficacy in developing animals, but potential neurotoxicity of these drugs is unexplored. To address this gap, we treated postnatal day (P)7 Sprague-Dawley rats with BRV (20, 40, 80 mg/kg) and PER (0.1, 0.9, 2.7 mg/kg), and assessed the induction of cell death across a range of vulnerable brain regions 24 h after exposure. Cell death was assessed using pathogreen staining. In each of the regions examined (dorsal striatum, nucleus accumbens, motor cortex, cingulate cortex, lateral thalamus, septum, hippocampus), VPA, which served as a positive control, significantly increased cell death as measured by the numer of pathogreen positive cells. By contrast, neither BRV nor PER increased the number of pathogreen positive cells in any region examined. Our results suggest that BRV and PER may have a positive safety profile-at least with respect to acute induction of cell death - and therefore may offer a safer option for the treatment of early life seizures.

Episodes of acute methotrexate-related neurotoxicity linked to compromised long-term neurocognitive function.

Methotrexate is a critical component of curative chemotherapy for pediatric acute lymphoblastic leukemia (ALL), but is associated with neurotoxicity. Information on long-term outcomes following an acute neurotoxic event is limited. Therefore, this report compares neurocognitive performance more than 12months post diagnosis (mean=4years) between ALL patients with (n=25) and without (n=146) a history of acute neurotoxicity. Compared to children with no documented on-treatment neurotoxic event, children who experienced a neurotoxic event during treatment exhibited poorer performance on measures of fine motor function (p=.02) and attention (p=.02). Children with ALL who experience acute neurotoxicity may be candidates for early neuropsychological screening and intervention.

The protective effects of fermented vinegar on acute alcohol-induced hepatic and neuro-toxicity by regulating ethanol metabolism and gut microbiota

The protective effects of fermented vinegar on acute alcohol-induced hepatic and neuro-toxicity by regulating ethanol metabolism and gut microbiota

Highly Porous Cellulose-Based Carbon Fibers as Effective Adsorbents for Chlorpyrifos Removal: Insights and Applications

The extensive utilization of the organophosphate pesticide chlorpyrifos, combined with its acute neurotoxicity, necessitates the development of effective strategies for its environmental removal. While numerous methods have been explored for chlorpyrifos removal from water, adsorption is the most promising. We investigated the potential of two cellulose-derived porous carbons as adsorbents for chlorpyrifos removal from water, prepared by either CO2 or H2O activation, resulting in similar morphologies and porosities but different amounts of heteroatom functionalities. The kinetics of batch adsorption removal from water fits well with the pseudo-first-order and pseudo-second-order kinetic models for both materials. The Freundlich, Langmuir, Dubinin–Radushkevich, and Sips isotherm models described the process of chlorpyrifos adsorption very well in all investigated cases. The maximum adsorption capacity determined from the Sips isotherm model gave values of 80.8 ± 0.1 mg g−1 and 132 ± 3 mg g−1 for the H2O and CO2 activated samples, respectively, reflecting the samples’ differences in heteroatom functionalities. Additionally, the application of either adsorbent led to reduced toxicity levels in all tested samples, implying that no harmful by-products were generated during adsorption. Comparative analysis with the existing literature further validates the study’s findings, suggesting the efficacy and applicability of cellulose-based porous carbons for sustainable chlorpyrifos remediation.

Outcomes of Stereotactic Radiation Therapy Versus Fractionated Radiation Therapy in 44 Dogs With Pituitary Masses: A Multi-Institutional Retrospective Study (2016-2022).

Although canine pituitary masses (PM) are increasingly treated with stereotactic radiotherapy (SRT), historical literature supports superior outcomes with conventional full-course fractionated radiation therapy (FRT). A multi-institutional retrospective study was performed, including dogs with PM treated from 2016 to 2022 with SRT (total dose 30 or 35 Gy in 5 daily fractions) or FRT (total dose 50-54 Gy in 19-20 daily fractions). The influence of potential prognostic/predictive factors was assessed, including pituitary: brain height, pituitary: brain volume, sex, age and endocrine status (functional [F] vs. nonfunctional [NF] PM). Forty-four dogs with PM were included (26 F, 14 NF, 4 unknown). All patients completed protocols as scheduled (SRT = 27, FRT = 17) and two dogs had suspected Grade 1 acute neurotoxicity. During the first 6 months after RT, 5/27 (19%) dogs treated with SRT (4 F, 1 NF) and 3/17 (18%) dogs treated with FRT (all F) died or were euthanised because of progressive neurologic signs. The overall median survival time was 608 days (95% CI, 375-840 days). Young age at the time of treatment was significant for survival (p = 0.0288); the overall median survival time was 753 days for dogs <9 years of age (95% CI, 614-892 days) and 445 days for dogs ≥9 years of age (95% CI, 183-707 days). Survival time was not associated with treatment type or any other factor assessed herein. A prospective study using standardised protocols would further validate the results of the present study and potentially elucidate the predictors of early death.

Preventing acute neurotoxicity of CNS therapeutic oligonucleotides with the addition of Ca2+ and Mg2+ in the formulation.

Oligonucleotide therapeutics (ASOs and siRNAs) have been explored for modulation of gene expression in the central nervous system (CNS), with several drugs approved and many in clinical evaluation. Administration of highly concentrated oligonucleotides to the CNS can induce acute neurotoxicity. We demonstrate that delivery of concentrated oligonucleotides to the CSF in awake mice induces acute toxicity, observable within seconds of injection. Electroencephalography (EEG) and electromyography (EMG) in awake mice demonstrated seizures. Using ion chromatography, we show that siRNAs can tightly bind Ca2+ and Mg2+ up to molar equivalents of the phosphodiester (PO)/phosphorothioate (PS) bonds independently of the structure or phosphorothioate content. Optimization of the formulation by adding high concentrations (above biological levels) of divalent cations (Ca2+ alone, Mg2+ alone, or Ca2+ and Mg2+) prevents seizures with no impact on the distribution or efficacy of the oligonucleotide. The data here establishes the importance of adding Ca2+ and Mg2+ to the formulation for the safety of CNS administration of therapeutic oligonucleotides.

Quinofumelin (Pesticides).

Food Safety Commission of Japan (FSCJ) conducted a risk assessment of quinofumelin (CAS No. 861647-84-9), a quinoline fungicide, based on submitted documents. The data used in the assessment are fate in plants (including paddy rice and tomatoes), residues in crops, fate in livestock (goats and chickens), residues in livestock products, fate in animals (rats), subacute toxicity (rats, mice, and dogs), chronic toxicity (dogs), combined chronic toxicity/carcinogenicity (rats), carcinogenicity (mice), acute neurotoxicity (rats), subacute neurotoxicity (rats), two-generation reproductive toxicity (rats), developmental toxicity (rats and rabbits), and genotoxicity. FSCJ specified an acceptable daily intake (ADI) of 0.03 mg/kg bw per day, and consequently specified an acute reference dose (ARfD) of 0.3 mg/kg bw per day after applying a safety factor of 100 based on the NOAEL.

Long-term follow-up of ARI0002h (cesnicabtagene autoleucel), an academic point-of-care B-cell maturation antigen (BCMA)-directed chimeric antigen receptor (CAR) T-cell strategy: Activity and safety after fractionated initial therapy and booster dose in 60 patients with relapsed/refractory multiple myeloma (RRMM).

7544 Background: The activity and safety ofARI0002h, an academic autologous CAR T-cell product with a humanized single chain variable fragment targeting BCMA has been reported in a pilot multicenter clinical trial (CARTBCMA-HCB-01) treating 30 patients (pts) with RRMM (NCT04309981) (Oliver-Caldés, Lancet Onc 2023). Here, we describe results of the final cohort of 60 pts with longer follow-up. Methods: Patients aged 18-75 years old with RRMM were eligible if they had measurable disease, were refractory to the last line of treatment and received ≥2 prior regimens, including a proteasome inhibitor, an immunomodulatory drug and an anti-CD38 antibody. The target dose (3x106/kg CAR+cells) was administered in a fractionated manner (10%/30%/60%). A second dose of up to 3x106 CAR+ cells/kg was planned at least 3 months after the first dose in pts with any kind of response and no limiting side effects. Primary endpoints were overall response rate (ORR) within the first 3 months and rate of cytokine release syndrome (CRS) and/or neurotoxicity in the first 30 days. Bone marrow minimal residual disease (MRD) was analyzed by next-generation flow at a sensitivity of 10-6. Results: As of December 18th 2023, 72 pts with RRMM were screened, 69 underwent apheresis and 61 received LD, with 60 pts finally receiving ARI0002h. The ORR in the first 3 months was 95% (≥ very good partial response (VGPR) in 77%). Median time to first response was one month. Responses deepened over time, achieving 58% complete response (CR) (55% stringent CR). MRD-negative rates on evaluable samples on days 28 and 100 were 98% and 96%, respectively. With a median follow-up of 24 months (95%CI 9.8-39.9), estimated median progression-free survival (PFS) was 20 months (95% CI 13.2-26.8). Median overall survival (OS) was not reached with OS rate at 24 months of 63%. CRS was observed in 90% with 5% grades ≥ 3. Median time to CRS was 7 days (1-14) with a median duration of 4.5 days. Mild acute neurotoxicity was reported in only 2 pts (3%) with no late neurologic events. 6 pts (10%) developed a macrophage activation syndrome (4 grades 1-2, 1 grade 3, 1 grade 5). Seven patients (11.7%) developed second primary malignancies after ARI0002h infusion, including 3 skin cancers (one in situ melanoma), 3 solid tumors and one acute myeloid leukemia. 80% (44 out of 5) eligible pts had already received the booster dose, with no relevant toxicities. Median time after first infusion was 4.4 months. Response was evaluable in 42 pts; 45% (n=19) were already in sCR, 29% (n=12) maintained the response and 26% (n=11) improved the response. Conclusions: Results from 60 pts and a longer follow-up confirm the safety profile and the deep and durable responses obtained after ARI0002h infusion. Clinical trial information: NCT04309981 .

Oxaliplatin-induced Acute Neurotoxicity Recovers Between Repeat Infusion Cycles: An Axonal Excitability Repeated Multiple Measurements Study.

Oxaliplatin, a platinum-based chemotherapy used in the treatment of colorectal cancer, induces acute neurotoxicity following infusion. The aim of this study was to establish whether alterations in axonal excitability develop progressively with higher cumulative doses and whether there is a recovery in motor axons after each cycle of treatment. Twenty consecutive patients with a colorectal cancer diagnosis, referred from the Oncology Department of Aretaieion Hospital of Athens, were enrolled in this study between October 2018 and May 2019. None of the participants had diabetes, alcohol abuse, known neuropathy or were previously treated with another neo-adjuvant therapy. Threshold Tracking techniques and Qtrac software were used for assessing axonal excitability in motor axons. Excitability recordings were undertaken before and immediately after the end of oxaliplatin infusion. Statistically significant changes were found (p<0.01) in axonal excitability (relative refractory period, refractoriness at 2 ms and 2.5 ms, sub-excitability and super-excitability) before and after oxaliplatin infusion. No statistically significant changes (p>0.05) were found in threshold electrotonus and strength-duration parameters before and after oxaliplatin infusion. We also did not find statistically significant differences (p>0.05) between means of excitability parameters before infusion at each cycle. Our study confirms oxaliplatin-induced acute neurotoxicity following infusion and suggests that motor axons recover between repeat infusion cycles.

Advancements in bioscavenger mediated detoxification of organophosphorus poisoning.

Organophosphorus compounds, widely used in agriculture and industry, pose a serious threat to human health due to their acute neurotoxicity. Although traditional interventions for organophosphate poisoning are effective, they often come with significant side effects. This paper aims to evaluate the potential of enzymes within biological organisms as organophosphorus bioclearing agents. It analyses the technical challenges in current enzyme research, such as substrate specificity, stereoselectivity, and immunogenicity, while exploring recent advancements in the field. A comprehensive review of literature related to detoxifying enzymes or proteins was conducted. Existing studies on organophosphorus bioclearing agents were summarised, elucidating the biological detoxification mechanisms, with a particular focus on advancements in protein engineering and novel delivery methods. Current bioclearing agents can be categorised into stoichiometric and catalytic bioclearing agents, both of which have shown some success in preventing organophosphate poisoning. Technological advancements have significantly improved various properties of bioclearing agents, yet challenges remain, particularly in substrate specificity, stereoselectivity, and immunogenicity. Future research will focus on expanding the substrate spectrum, enhancing catalytic efficiency, prolonging in vivo half-life, and developing convenient administration methods. With the progression of clinical trials, bioclearing agents are expected to become widely used as a new generation of therapeutic organophosphate detoxifiers.

Favorable efficacy and reduced acute neurotoxicity by antisense oligonucleotides with 2′,4′-BNA/LNA with 9-(aminoethoxy)phenoxazine

An increasing number of antisense oligonucleotides (ASOs) have been approved for clinical use. However, improvements of both efficacy and safety in the central nervous system (CNS) are crucial for the treatment with CNS diseases. We aimed to overcome the crucial issues by our development of various gapmer ASOs with anovel nucleoside derivative including a 2',4'-BNA/LNA with9-(aminoethoxy)phenoxazine (BNAP-AEO). The various gapmer ASOs with BNAP-AEO were evaluated for thermal stability, invitro and invivo efficacy, and acute CNS toxicity. Thermal stability analysis of the duplexes with their complementary RNAs showed that ASOs with BNAP-AEO had a higher binding affinity than those without BNAP-AEO. Invitro assays, when transfected into neuroblastoma cell lines, demonstrated that ASOs with BNAP-AEO, had a more efficient gene silencing effect than those without BNAP-AEO. Invivo assays, involving intracerebroventricular injections into mice, revealed ASOs with BNAP-AEO potently suppressed gene expression in the brain. Surprisingly, the acute CNS toxicity in mice, as assessed through open field tests and scoring systems, was significantly lower for ASOs with BNAP-AEO than for those without BNAP-AEO. This study underscores the efficient gene-silencing effect and low acute CNS toxicity of ASOs incorporating BNAP-AEO, indicating the potential for future therapeutic applications.

Silica nanoparticles: Chemical synthesis and acute neurotoxic study in Wistar rat

Objectives: In this study, silica nanoparticles (SNPs) were synthesized through the sol-gel approach, characterized using X-ray diffraction and examined for their potential to induce acute neurotoxicity in rats. Materials and Methods: The average particle size was confirmed to be 110 ± 5 nm through scanning electron microscope (SEM) analysis. An intraperitoneal injection of 25, 50, and 100 mg/kg of SNPs was administered twice over 24 h to rats, followed by autopsies 24 h later. Brain regions (cerebellum, frontal cortex, hippocampus, and corpus striatum) were analyzed for changes in antioxidant enzyme activity (superoxide dismutase [SOD], catalase [CAT], glutathione-s-transferase [GST], glutathione peroxidase [GPx], glutathione reductase [GR] and thiobarbituric acid reactive substance [TBARS] levels) and neurotransmitter concentrations (acetylcholinestrase [AChE], dopamine, serotonin and nor-epinephrine). Results: The activity of SOD, CAT and GST increased significantly in the high dose group, specifically in the hippocampus and corpus striatum. No significant change in GR activity was observed in any dose group. Significant alterations were observed in neurotransmitter concentrations in hippocampus and corpus striatum in rats treated with high dose. Conclusion: Overall, the outcomes derived from our research suggest that SNPs lead to dose-dependent oxidative harm and neurotoxic effects in different regions of brain.

Acetylcholinesterase Inhibition in Rats and Humans Following Acute Fenitrothion Exposure Predicted by Physiologically Based Kinetic Modeling-Facilitated Quantitative In Vitro to In Vivo Extrapolation.

Worldwide use of organophosphate pesticides as agricultural chemicals aims to maintain a stable food supply, while their toxicity remains a major public health concern. A common mechanism of acute neurotoxicity following organophosphate pesticide exposure is the inhibition of acetylcholinesterase (AChE). To support Next Generation Risk Assessment for public health upon acute neurotoxicity induced by organophosphate pesticides, physiologically based kinetic (PBK) modeling-facilitated quantitative in vitro to in vivo extrapolation (QIVIVE) approach was employed in this study, with fenitrothion (FNT) as an exemplary organophosphate pesticide. Rat and human PBK models were parametrized with data derived from in silico predictions and in vitro incubations. Then, PBK model-based QIVIVE was performed to convert species-specific concentration-dependent AChE inhibition obtained from in vitro blood assays to corresponding in vivo dose-response curves, from which points of departure (PODs) were derived. The obtained values for rats and humans were comparable with reported no-observed-adverse-effect levels (NOAELs). Humans were found to be more susceptible than rats toward erythrocyte AChE inhibition induced by acute FNT exposure due to interspecies differences in toxicokinetics and toxicodynamics. The described approach adequately predicts toxicokinetics and acute toxicity of FNT, providing a proof-of-principle for applying this approach in a 3R-based chemical risk assessment paradigm.

MRI brain findings of Abamectin toxic encephalopathy: a case report with review of literature.

Abamectin is an insecticidal/miticidal compound derived from the soil bacterium "Streptomyces avermitilis". Abamectin toxicity in humans is very rare. We present a case of acute neurotoxicity induced by Abamectin, showcasing distinctive MRI brain findings in a 33-year-old female who exhibited a favourable recovery with the aid of supportive care. In a patient with known exposure to toxins, even with a lack of knowledge of the specific type or class of toxin, recognition of anatomical distribution of lesions on brain MRI and their characteristic appearance can help exclude other causes of neurologic impairment and aid in timely management.

Development of a Novel Methotrexate-Related Neurotoxicity Risk Score in Pediatric Acute Leukemia: A Report from the Reducing Ethnic Disparities in Acute Leukemia (REDIAL) Consortium

Introduction: Methotrexate (MTX) chemotherapy is a critical component of contemporary pediatric acute lymphoblastic leukemia (ALL) treatment regimens. However, intrathecal (IT) and intravenous (IV) MTX can result in dose-limiting acute neurotoxicity, which has been associated with an increased risk of ALL relapse. Emerging data suggest the likelihood of MTX neurotoxicity is elevated in Latino patients and those diagnosed at older age or treated with high-dose MTX (&amp;gt; 1g/m2); however, there are no established methods for identifying those at greatest risk. Therefore, we used machine learning algorithms to develop a neurotoxicity risk prediction model in a diverse, multi-ethnic cohort of pediatric patients with ALL. Methods: The Reducing Ethnic Disparities in Acute Leukemia (REDIAL) Consortium enrolls patients diagnosed with ALL from six participating treatment centers in the Southwestern U.S. This interim analysis included pediatric patients (age &amp;lt;20 years) diagnosed and treated for ALL at Texas Children's Hospital (2005-2019). Individuals diagnosed with infant leukemia (&amp;lt;1 year of age) and/or with preexisting neurologic disease were excluded. We retrospectively reviewed medical records to identify clinical, sociodemographic, and Common Terminology Criteria for Adverse Events (CTCAE) grading of toxicities such as transaminitis and hyperbilirubinemia. Address information at the time of diagnosis was geocoded and aligned to the American Community Survey at the census-tract level to define residential social determinants of health (SDOH). Cases of MTX neurotoxicity were identified by manual review of records for any patient who underwent brain MRI imaging during ALL therapy. Cases of acute MTX neurotoxicity were defined following Ponte di Legno Delphi Consensus definitions as neurologic episodes (e.g., seizure, stroke-like symptoms, altered mental status) occurring within 21 days of IV or IT MTX with no other identifiable underlying cause. Demographic and clinical factors were compared between cases of acute MTX-related neurotoxicity and controls using logistic regression models to estimate odds ratios (OR) and 95% confidence intervals (CI). The dataset was randomly split with 80% of observations used in the training stage and 20% reserved for testing. Random forest algorithms with boosting and down sampling were grown using 5-repeat, 10-fold cross validation to optimize tuning parameters. Model performance was evaluated using test and train error rate, area under the curve of the receiver operating characteristic (AUC-ROC), sensitivity, and specificity. Results: A total of 770 patients were eligible and met inclusion criteria. Patients were a mean age of 7.0 years at diagnosis, 54% male, 55% Latino and 45% treated with high-risk protocols. Acute MTX neurotoxicity developed in 134 (17.4%) of patients. In logistic regression models, neurotoxicity odds increased with older age at diagnosis (OR = 1.20, 95% CI: 1.15-1.26), Latino ethnicity (OR = 2.14, 95% CI: 1.38-3.38), treatment on high-risk protocols (OR=4.32, 95% CI: 2.75-6.99), obesity at diagnosis (vs normal weight, OR = 2.54, 95% CI: 1.51-4.22), and CTCAE grade 3 or higher transaminitis (OR = 2.02, 95% CI: 1.03-4.47) or hyperbilirubinemia (OR = 4.71, 95% CI: 1.21-18.41) during induction therapy. Overall, the random forest machine learning algorithm resulted in an AUC-ROC of 0.79 with an error rate of 0.26 in both the training and test sets. The overall sensitivity was 0.71, with a specificity of 0.67. Recognizing ethnic disparities in MTX neurotoxicity may exist, random forest models were generated stratifying on ethnicity (Figure 1), which yielded slightly better performance in non-Latino patients (AUC-ROC = 0.77) than Latino patients (AUC-ROC = 0.73). Conclusions: This interim analysis of data from the REDIAL Consortium demonstrates the potential utility of machine learning algorithms to develop a novel risk prediction model for acute MTX-related neurotoxicity. Efforts to expand this work within the larger REDIAL cohort (n&amp;gt;2,000) and incorporate inherited genetic data are ongoing. Ultimately, this work may inform efforts to preemptively risk-stratify patients and deliver targeted interventions to vulnerable individuals.

Mechanochemical synthesis and anticonvulsant activity of 3-aminopyrrolidine-2,5-dione derivatives

A series of 3-aminopyrrolidine-2,5-dione derivatives was synthesized and tested for anticonvulsant activity. Succinimide derivatives were obtained from a simple solvent-based reaction and a mechanochemical aza-Michael reaction of maleimide or its N-substituted derivatives with selected amines. The structure of the compounds was confirmed by spectroscopic methods (NMR, FT-IR, HPLC, ESI-MS, EA and XRD for four compounds). The cytotoxic activity of the succinimide derivatives was evaluated using HepG2 cells for hepatocytotoxicity and SH-SY5Y cells for neurocytotoxicity. None of the studied compounds showed hepatocytotoxicity and two showed neurocytotoxicity. Initial anticonvulsant screening was performed in mice using the psychomotor seizure test (6 Hz, 32 mA). The selected compounds were evaluated in the following acute models of epilepsy: the maximal electroshock test, psychomotor seizure test (6 Hz, 44 mA), subcutaneous pentylenetetrazole seizure test, and acute neurotoxicity (rotarod test). The most active compound 3-((4-chlorophenyl)amino)pyrrolidine-2,5-dione revealed antiseizure activity in all seizure models (including pharmacoresistant seizures) and showed better median effective doses (ED50) and protective index values than the reference compound, ethosuximide. Furthermore, 3-(benzylamino)pyrrolidine-2,5-dione and 3-(phenylamino)pyrrolidine-2,5-dione exhibited antiseizure activity in the 6 Hz and MES tests, and 3-(butylamino)−1-phenylpyrrolidine-2,5-dione and 3-(benzylamino)−1-phenylpyrrolidine-2,5-dione exhibited antiseizure activity in the 6 Hz test. All active compounds demonstrated low in vivo neurotoxicity in the rotarod test and yielded favourable protective indices.