Current protocols in toxicology最新文献_第6页

Evaluation of Marijuana Compounds on Neuroimmune Endpoints in Experimental Autoimmune Encephalomyelitis 大麻化合物对实验性自身免疫性脑脊髓炎神经免疫终点的影响

Current protocols in toxicology Pub Date : 2018-02-21 DOI: 10.1002/cptx.43

Barbara L. F. Kaplan

引用次数: 6

Determination of Mycophenolic Acid and Mycophenolic Acid Glucuronide Using Liquid Chromatography Tandem Mass Spectrometry (LC/MS/MS) 液相色谱串联质谱法(LC/MS/MS)测定霉酚酸和霉酚酸葡糖苷

Current protocols in toxicology Pub Date : 2018-02-21 DOI: 10.1002/cptx.42

Uttam Garg, Ada Munar, Clinton Frazee

{"title":"Determination of Mycophenolic Acid and Mycophenolic Acid Glucuronide Using Liquid Chromatography Tandem Mass Spectrometry (LC/MS/MS)","authors":"Uttam Garg, Ada Munar, Clinton Frazee","doi":"10.1002/cptx.42","DOIUrl":"10.1002/cptx.42","url":null,"abstract":"Mycophenolic acid (MPA) is an immunosuppressant that is used in renal, liver, and heart transplantation. Due to its narrow therapeutic range, monitoring of MPA levels is essential to avoid toxicity and organ rejection. Although immunoassays are available for the determination of MPA, due to their higher specificity, mass spectrometry methods are preferred. In this unit, we describe a liquid chromatography tandem mass spectrometry (LC/MS/MS) method utilizing positive ionization electrospray and multiple reaction monitoring (MRM) for the quantification of levels of MPA and its conjugate MPA glucuronide (MPAG). Blood collected in a plain, EDTA, or heparin-containing tube is centrifuged to separate the serum or plasma. Proteins are precipitated using a solution containing zinc sulfate and acetonitrile that has been spiked with deuterated internal standards. The resulting protein-free supernatant is injected into the LC/MS/MS system for analysis. The chromatography involves the use of a C18 column and ammonium acetate/water/formic acid and ammonium acetate/methanol/formic acid mobile phases. Quantification of MPA and MPAG levels is achieved by comparing the MRM peak area ratios of analytes and internal standards, consisting of specific precursor/product pairs, with those of calibrators at various concentrations. Calibration curves are constructed from the MRM peak area ratios of calibrators and internal standards versus concentration. © 2018 by John Wiley & Sons, Inc.","PeriodicalId":72743,"journal":{"name":"Current protocols in toxicology","volume":"75 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cptx.42","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35890116","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: 4

Use of Human Embryoid Bodies for Teratology 人类胚胎样体在畸形学中的应用

Current protocols in toxicology Pub Date : 2018-02-21 DOI: 10.1002/cptx.38

Anthony Flamier, Supriya Singh, Theodore P. Rasmussen

引用次数: 2

Phenotypic and Functional Evaluation of Hematopoietic Stem and Progenitor Cells in Toxicology of Heavy Metals 造血干细胞和祖细胞在重金属毒理学中的表型和功能评价

Current protocols in toxicology Pub Date : 2018-02-21 DOI: 10.1002/cptx.41

Qian Li, Zhengli Yang, Yifan Zhao, Xiaodong Jia, Zhijun Zhou, Yubin Zhang

引用次数: 7

A Real-Time Image-Based Approach to Distinguish and Discriminate Apoptosis from Necrosis 基于实时图像的细胞凋亡与坏死的鉴别方法

Current protocols in toxicology Pub Date : 2018-02-21 DOI: 10.1002/cptx.39

Asha Lekshmi, Shankara Narayanan Varadarajan, Santhik Subhasingh Lupitha, Mydhily Nair, Aneesh Chandrasekharan, T.R. Santhoshkumar

{"title":"A Real-Time Image-Based Approach to Distinguish and Discriminate Apoptosis from Necrosis","authors":"Asha Lekshmi, Shankara Narayanan Varadarajan, Santhik Subhasingh Lupitha, Mydhily Nair, Aneesh Chandrasekharan, T.R. Santhoshkumar","doi":"10.1002/cptx.39","DOIUrl":"10.1002/cptx.39","url":null,"abstract":"Recent cell biology studies reveal that a cell can die through multiple pathways via distinct signaling mechanisms. Among these, apoptosis and necrosis are two distinct cell death pathways, and their detection and discrimination is vital in the drug discovery process and in understanding diverse biological processes. Although sensitive assays for apoptosis and necrosis are available, it is extremely difficult to adapt any of these methods to discriminate apoptosis-inducing stimuli from necrosis-inducing stimuli because of the acquisition of secondary necrosis by apoptotic cells when they are not phagocytosed. Essentially, any assay for discriminating apoptosis and necrosis needs to be carried out in real-time kinetic mode. Caspase 3 or 7 activation is observed in the majority of apoptotic cell death. Similarly, the absence of caspase 3/7 activation and cell membrane leakage are the two prominent indicators for necrotic cell death or necroptosis. The programmed form of necrosis, called pyroptosis, is also accompanied by membrane leakage and most often associated with activation of specific caspases such as caspase 1, 4, or 11, but not through caspase 3/7 activation. Here, a robust and sensitive real-time method is described to distinguish and discriminate apoptosis from necrosis. The assay utilizes stable integration of a genetically encoded fluorescence resonance energy transfer (FRET) probe for caspase 3/7 activation and the mitochondrion-targeted DsRed to identify necrotic cells. Caspase activation is determined by cleavage of the FRET probe; loss of soluble FRET probe with retention of mitochondrial red fluorescence indicates necrosis. This unit describes an important protocol for the generation of sensor cells expressing both probes, followed by detailed analysis of apoptosis and necrosis by microscopy imaging, confocal imaging, high-throughput imaging, and flow cytometry. © 2018 by John Wiley & Sons, Inc.","PeriodicalId":72743,"journal":{"name":"Current protocols in toxicology","volume":"75 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cptx.39","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35891191","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: 3

Preparation of Human Primary Colon Tissue-Derived Organoid Using Air Liquid Interface Culture 气液界面培养制备人原代结肠组织类器官

Current protocols in toxicology Pub Date : 2018-02-21 DOI: 10.1002/cptx.40

Tatsuya Usui, Masashi Sakurai, Koji Umata, Hideyuki Yamawaki, Takashi Ohama, Koichi Sato

{"title":"Preparation of Human Primary Colon Tissue-Derived Organoid Using Air Liquid Interface Culture","authors":"Tatsuya Usui, Masashi Sakurai, Koji Umata, Hideyuki Yamawaki, Takashi Ohama, Koichi Sato","doi":"10.1002/cptx.40","DOIUrl":"10.1002/cptx.40","url":null,"abstract":"In vitro analysis of intestinal epithelium has been hindered by a lack of suitable culture systems useful for gastrointestinal research. To overcome the problem, an air liquid interface (ALI) method using a collagen gel was established to culture three-dimensional primary cells containing both primary epithelial and mesenchymal components from mouse gastrointestinal tissues. ALI organoids accurately recapitulate organ structures, multilineage differentiation, and physiology. Since ALI organoids from human tissues have not been produced, we modified the previous protocol for mouse ALI organoid culture to establish the culture system of ALI organoids from normal and tumor colorectal tissues of human patients. The current unit presents a protocol for preparation of the ALI organoid culture from normal and tumor colorectal tissues of human patients. ALI organoid culture from human tissues might be useful for examining not only resistance to chemotherapy in a tumor microenvironment but also toxic effects on organoids. © 2018 by John Wiley & Sons, Inc.","PeriodicalId":72743,"journal":{"name":"Current protocols in toxicology","volume":"75 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cptx.40","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35891188","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: 19

In Vivo Measurement of Intragastric Pressure with a Rubber Balloon in the Anesthetized Rat 橡胶球囊在体测量麻醉大鼠胃内压

Current protocols in toxicology Pub Date : 2018-02-16 DOI: 10.1002/0471140856.tx2112s57

Zoltán S. Zádori, Klára Gyires

引用次数: 3

Methods to Detect Protein Glutathionylation 方法检测蛋白谷胱甘肽化

Current protocols in toxicology Pub Date : 2018-02-16 DOI: 10.1002/0471140856.tx0617s57

Robyn L. Poerschke, Kristofer S. Fritz, Christopher C. Franklin

{"title":"Methods to Detect Protein Glutathionylation","authors":"Robyn L. Poerschke, Kristofer S. Fritz, Christopher C. Franklin","doi":"10.1002/0471140856.tx0617s57","DOIUrl":"10.1002/0471140856.tx0617s57","url":null,"abstract":"Glutathionylation is a posttranslational modification that results in the formation of a mixed disulfide between glutathione and the thiol group of a protein cysteine residue. Glutathionylation of proteins occurs via both nonenzymatic mechanisms involving thiol/disulfide exchange and enzyme-mediated reactions. Protein glutathionylation is observed in response to oxidative or nitrosative stress and is redox-dependent, being readily reversible under reducing conditions. Such findings suggest that glutathionylation plays an important role in mediating redox-sensitive signaling. Indeed, glutathionylation can affect protein function by altering activity, protein-protein interactions, and ligand binding. Glutathionylation may also serve to prevent cysteine residues from undergoing irreversible oxidative modification. Thus, determining the ability of a given protein to become glutathionylated can provide insight into its redox regulation and putative role in dictating cellular response to oxidative and nitrosative stress. Methods to measure protein glutathionylation using immunoblotting and mass spectrometry are described. Curr. Protoc. Toxicol. 57:6.17.1-6.17.18. © 2013 by John Wiley & Sons, Inc.","PeriodicalId":72743,"journal":{"name":"Current protocols in toxicology","volume":"57 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-02-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/0471140856.tx0617s57","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32103786","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: 3

Overview of Neurotoxicology 神经毒理学综述

Current protocols in toxicology Pub Date : 2018-02-13 DOI: 10.1002/cptx.36

Lucio G. Costa

引用次数: 9

Isolation, Cryopreservation, and Immunophenotyping of Human Peripheral Blood Mononuclear Cells 人外周血单个核细胞的分离、冷冻保存和免疫表型分析

Current protocols in toxicology Pub Date : 2018-02-13 DOI: 10.1002/cptx.31

Fredine T. Lauer, Jesse L. Denson, Scott W. Burchiel

引用次数: 17