Journal of pharmacological and toxicological methods最新文献

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A novel, flexible, and accessible method for the ex vivo induction and quantification of excitotoxicity. 一种新颖的、灵活的、可接近的体外诱导和定量兴奋毒性的方法。

Journal of pharmacological and toxicological methods Pub Date : 2025-10-06 DOI: 10.1016/j.vascn.2025.108401

Samantha J Carew, Christiana M Kennedy, Meghan L Greenland, Matthew P Parsons

{"title":"A novel, flexible, and accessible method for the ex vivo induction and quantification of excitotoxicity.","authors":"Samantha J Carew, Christiana M Kennedy, Meghan L Greenland, Matthew P Parsons","doi":"10.1016/j.vascn.2025.108401","DOIUrl":"https://doi.org/10.1016/j.vascn.2025.108401","url":null,"abstract":"Excitotoxicity is a key driver of neuronal death in diverse brain conditions, yet most toxicity assays rely on in vitro models that remove cells from their complex native environment within the brain parenchyma. Here, we present a novel ex vivo method to quantify N-methyl-d-aspartate (NMDA)-induced excitotoxicity using acute brain slices from male and female young adult c57bl/6 and aged B6129SF2J mice, similar to those used for conventional electrophysiological recordings. Acute hippocampal slices were recovered in an N-methyl-D-glucamine (NMDG)-based recovery solution, then treated with low-magnesium aCSF containing the co-agonist glycine to promote receptor activation, with or without exogenous NMDA. Following treatment, slices were fixed, cryoprotected, and cryosectioned to 20 μm for immunohistochemistry. Apoptotic cell death was assessed by staining for cleaved caspase-3, and was combined with the percentage of dead space to calculate a toxicity index for overall excitotoxic cell death. Importantly, exposure to low-magnesium aCSF with glycine alone was sufficient to elevate active caspase-3 levels, an effect that was further enhanced by exogenous NMDA application and prevented by NMDAR antagonism. Our ex vivo method largely preserves the cytoarchitecture and local microenvironment of brain tissue, enabling the assessment of cell-specific vulnerabilities to excitotoxic damage in select brain regions at defined ages. It is particularly well-suited for use in neurodegenerative disease models, where excitotoxic susceptibility may evolve over time. In all, the approach described here provides a reliable and accessible alternative to dissociated cell cultures, bridging the gap between in vitro and in vivo systems for studying glutamate-induced cell death.","PeriodicalId":94102,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":" ","pages":"108401"},"PeriodicalIF":0.0,"publicationDate":"2025-10-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145254150","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Therapeutic drug monitoring of busulfan and cyclophosphamide in hematologic malignancies: An LC-MS/MS development, validation and application. 丁硫凡和环磷酰胺在血液恶性肿瘤中的治疗药物监测：LC-MS/MS开发、验证和应用。

Journal of pharmacological and toxicological methods Pub Date : 2025-09-01 Epub Date: 2025-08-05 DOI: 10.1016/j.vascn.2025.108389

Jinxingyi Wang, Ruyu Tao, Hanshuai Hu, Xue Lan, Jiejie Gao, Yanping Guan, Qian Liu

{"title":"Therapeutic drug monitoring of busulfan and cyclophosphamide in hematologic malignancies: An LC-MS/MS development, validation and application.","authors":"Jinxingyi Wang, Ruyu Tao, Hanshuai Hu, Xue Lan, Jiejie Gao, Yanping Guan, Qian Liu","doi":"10.1016/j.vascn.2025.108389","DOIUrl":"10.1016/j.vascn.2025.108389","url":null,"abstract":"Background: Busulfan and cyclophosphamide (Bu-Cy) is a widely used conditioning regimen for hematopoietic stem cell transplantation (HSCT) in the treatment of hematologic malignancies. To explore the exposure-efficiency relationships of Bu-Cy regimen and evaluate the necessity of relative therapeutic drug monitoring (TDM), we developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to simultaneously quantify the busulfan and cyclophosphamide in human plasma.Methods: A simple LC-MS/MS bioassay for simultaneous determination of busulfan and cyclophosphamide using isotope dilution internal standardization has been established. Plasma samples were subjected to protein precipitation prior to their injection into a column for subsequent separation (Thermo Accucore aQ C18, 50 × 2.1 mm, 2.6 μm). Positive electrospray ionization (ESI) mode with selected reaction monitoring (SRM) was used for analytes determination.Results: Busulfan and cyclophosphamide could be measured within 3 min, with the calibration ranges of 80-4000 ng/mL for busulfan and 25-2000 ng/mL for cyclophosphamide. The validated method showed good sensitivity and linearity. Accuracies and precisions were less than 15 %. The results of stability showed that busulfan and cyclophosphamide were stable in the plasma under room temperature for 16 h, 4 °C for 24 h, -80 °C for 2 weeks, and after three freeze-thaw processes.Conclusion: The bioassay was simple, sensitive and successfully implemented in the realm of TDM for Bu-Cy, ultimately leading to improved dosing precision.","PeriodicalId":94102,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":" ","pages":"108389"},"PeriodicalIF":0.0,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144786281","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Corrigendum to "Best practice considerations for nonclinical in vivo cardiovascular telemetry studies in non-rodent species: Delivering high quality QTc data to support ICH E14/S7B Q&As" [J. Pharmacol. Toxicol. Methods 123 (2023) 107270]. 《非啮齿类动物非临床体内心血管遥测研究的最佳实践考虑：提供高质量的QTc数据以支持ICH E14/S7B Q&As》的更正[J]。杂志。Toxicol。方法123(2023)107270]。

Journal of pharmacological and toxicological methods Pub Date : 2025-09-01 Epub Date: 2025-09-16 DOI: 10.1016/j.vascn.2025.108392

Eric I Rossman, Todd A Wisialowski, Hugo M Vargas, Jean-Pierre Valentin, Michael G Rolf, Brian M Roche, Steve Riley, Michael K Pugsley, Jill Nicholsi, Dingzhou Li, Derek J Leishman, Robert B Kleiman, Andrea Greiter-Wilke, Gary A Gintant, Michael J Engwall, Annie Delaunois, Simon Authier

引用次数: 0