Current protocols in toxicology最新文献_第9页

Assessment of Bile Salt Export Pump (BSEP) Inhibition in Membrane Vesicles Using Radioactive and LC/MS-Based Detection Methods 利用放射性和LC/ ms检测方法评价胆汁盐出口泵(BSEP)对膜泡的抑制作用

Current protocols in toxicology Pub Date : 2017-02-01 DOI: 10.1002/cptx.15

Lisa D. Marroquin, Paul D. Bonin, Julie Keefer, Thomas Schroeter

{"title":"Assessment of Bile Salt Export Pump (BSEP) Inhibition in Membrane Vesicles Using Radioactive and LC/MS-Based Detection Methods","authors":"Lisa D. Marroquin, Paul D. Bonin, Julie Keefer, Thomas Schroeter","doi":"10.1002/cptx.15","DOIUrl":"10.1002/cptx.15","url":null,"abstract":"The bile salt export pump (BSEP, ABCB11) belongs to the ATP-binding-cassette superfamily of transporters and is predominately found in the liver. BSEP is an efflux transporter that plays a critical role in the secretion of bile salts into the bile. Inhibition of BSEP function by drugs can result in the buildup of bile salts in the liver and eventually leads to cholestasis and drug-induced liver injury (DILI). DILI is a major cause of withdrawal of drugs from the pharmaceutical market and accounts for >50% of acute liver failures. Therefore, early detection of BSEP inhibition by drugs can help to mitigate the possibility of BSEP-associated liver injury. This unit describes two assays that investigate the relationship between drug interference with BSEP function and liver injury using membrane vesicles prepared from Hi5 insect cells transfected with human BSEP. Comprehensive protocols for assessing BSEP inhibition in a 384-well format using radiolabeled and liquid chromatography/mass spectrometry (LC/MS)–based detection methods are described. © 2017 by John Wiley & Sons, Inc.","PeriodicalId":72743,"journal":{"name":"Current protocols in toxicology","volume":"71 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2017-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cptx.15","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45741597","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: 8

Preparation of Specific Compartments of the Lungs for Pathologic and Biochemical Analysis of Toxicologic Responses 制备用于毒理学反应病理和生化分析的特定肺室

Current protocols in toxicology Pub Date : 2017-02-01 DOI: 10.1002/cptx.18

Laura S. Van Winkle, Jacklyn S. Kelty, Charles G. Plopper

引用次数: 5

Fluorescence-Based Microplate Assays for In Vitro Assessment of Mitochondrial Toxicity, Metabolic Perturbation, and Cellular Oxygenation 荧光微孔板法测定线粒体毒性，代谢扰动和细胞氧化的体外评估

Current protocols in toxicology Pub Date : 2016-11-01 DOI: 10.1002/cptx.3

James Hynes, Conn Carey, Yvonne Will

引用次数: 8

Basic Techniques in Mammalian Cell Tissue Culture 哺乳动物细胞组织培养的基本技术

Current protocols in toxicology Pub Date : 2016-11-01 DOI: 10.1002/cptx.13

Katy Phelan, Kristin M. May

引用次数: 55

In Vivo Determination of Mitochondrial Function Using Luciferase-Expressing Caenorhabditis elegans: Contribution of Oxidative Phosphorylation, Glycolysis, and Fatty Acid Oxidation to Toxicant-Induced Dysfunction 用表达荧光素酶的秀丽隐杆线虫在体内测定线粒体功能:氧化磷酸化、糖酵解和脂肪酸氧化对毒物诱导功能障碍的贡献

Current protocols in toxicology Pub Date : 2016-08-01 DOI: 10.1002/cptx.10

Anthony L. Luz, Cristina Lagido, Matthew D. Hirschey, Joel N. Meyer

{"title":"In Vivo Determination of Mitochondrial Function Using Luciferase-Expressing Caenorhabditis elegans: Contribution of Oxidative Phosphorylation, Glycolysis, and Fatty Acid Oxidation to Toxicant-Induced Dysfunction","authors":"Anthony L. Luz, Cristina Lagido, Matthew D. Hirschey, Joel N. Meyer","doi":"10.1002/cptx.10","DOIUrl":"10.1002/cptx.10","url":null,"abstract":"Mitochondria are a target of many drugs and environmental toxicants; however, how toxicant-induced mitochondrial dysfunction contributes to the progression of human disease remains poorly understood. To address this issue, in vivo assays capable of rapidly assessing mitochondrial function need to be developed. Here, using the model organism Caenorhabditis elegans, we describe how to rapidly assess the in vivo role of the electron transport chain, glycolysis, or fatty acid oxidation in energy metabolism following toxicant exposure, using a luciferase-expressing ATP reporter strain. Alterations in mitochondrial function subsequent to toxicant exposure are detected by depleting steady-state ATP levels with inhibitors of the mitochondrial electron transport chain, glycolysis, or fatty acid oxidation. Differential changes in ATP following short-term inhibitor exposure indicate toxicant-induced alterations at the site of inhibition. Because a microplate reader is the only major piece of equipment required, this is a highly accessible method for studying toxicant-induced mitochondrial dysfunction in vivo. © 2016 by John Wiley & Sons, Inc.","PeriodicalId":72743,"journal":{"name":"Current protocols in toxicology","volume":"69 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cptx.10","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34331887","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: 27

Multielectrode Array (MEA) Assay for Profiling Electrophysiological Drug Effects in Human Stem Cell-Derived Cardiomyocytes 多电极阵列(MEA)分析分析电生理药物对人干细胞源性心肌细胞的影响

Current protocols in toxicology Pub Date : 2016-05-04 DOI: 10.1002/cptx.2

Mike Clements

{"title":"Multielectrode Array (MEA) Assay for Profiling Electrophysiological Drug Effects in Human Stem Cell-Derived Cardiomyocytes","authors":"Mike Clements","doi":"10.1002/cptx.2","DOIUrl":"10.1002/cptx.2","url":null,"abstract":"More relevant and reliable preclinical cardiotoxicity tests are required to improve drug safety and reduce the cost of drug development. Human stem cell-derived cardiomyocytes (hSC-CMs) provide a potential model for the development of superior assays for preclinical drug safety screening. One such hSC-CM assay that has shown significant potential for enabling more predictive drug cardiac risk assessment is the MEA assay. The Multi-electrode Array (MEA) assay is an electrophysiology-based technique that uses microelectrodes embedded in the culture surface of each well to measure fluctuations in extracellular field potential (FP) generated from spontaneously beating hSC-CMs. Perturbations to the recorded FP waveform can be used as an unbiased method of predicting the identity of ion channel(s) impacted on drug exposure. Here, a higher throughput MEA assay using hSC-CMs in 48-well MEA plates is described for profiling compound-induced effects on cardiomyocyte electrophysiology. Techniques for preparing hSC-CM monolayers in MEA plates and methods to contextualize MEA assay experimental results are also covered. © 2016 by John Wiley & Sons, Inc.","PeriodicalId":72743,"journal":{"name":"Current protocols in toxicology","volume":"68 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-05-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cptx.2","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34454940","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: 29

Investigating the Effects of Particulate Matter on House Dust Mite and Ovalbumin Allergic Airway Inflammation in Mice 颗粒物质对小鼠屋尘螨和卵清蛋白过敏性气道炎症的影响

Current protocols in toxicology Pub Date : 2016-05-04 DOI: 10.1002/cptx.5

Alejandro R. Castañeda, Kent E. Pinkerton

{"title":"Investigating the Effects of Particulate Matter on House Dust Mite and Ovalbumin Allergic Airway Inflammation in Mice","authors":"Alejandro R. Castañeda, Kent E. Pinkerton","doi":"10.1002/cptx.5","DOIUrl":"10.1002/cptx.5","url":null,"abstract":"Particulate matter (PM), a component of air pollution, has been shown to enhance allergen-mediated airway hypersensitivity and inflammation. Surprisingly, exposure to PM during the sensitization to allergen is sufficient to produce immunological changes that result in heightened inflammatory effects upon future allergen exposures (challenge) in the absence of PM. This suggests that PM has the ability to modulate the allergic immune response, thereby acting as an adjuvant by enhancing the immunological memory formed during the adaptive immune response; however, the mechanisms through which this occurs remain elusive. Establishing a reproducible animal model to study the PM-mediated immunotoxicological effects that enhance allergy, may provide insights to understand how air pollution activates the immune system and thereby modulates the pathophysiology of asthma. The basic protocol can be used to study various characteristics of air pollution, such as PM size, source, or chemical composition, to help elucidate how such features may affect the allergic response in a mouse model of asthma. Using a BALB/c model of acute exposure (14 days), mice are first sensitized with allergen and PM, and then subsequently challenged with allergen only. The endpoints of this basic protocol include the assessment of inflammation via cells recovered from broncho-alveolar lavage (BAL), histopathological analysis, gene expression profiles, and protein quantification of inflammatory markers. © 2016 by John Wiley & Sons, Inc.","PeriodicalId":72743,"journal":{"name":"Current protocols in toxicology","volume":"68 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-05-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cptx.5","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34453447","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: 15

Determination of Metabolic Viability and Cell Mass Using a Tandem Resazurin/Sulforhodamine B Assay 利用瑞唑脲/硫代丹胺B串联测定代谢活力和细胞质量

Current protocols in toxicology Pub Date : 2016-05-04 DOI: 10.1002/cptx.1

Filomena S.G. Silva, Irina G. Starostina, Vilena V. Ivanova, Albert A. Rizvanov, Paulo J. Oliveira, Susana P. Pereira

引用次数: 37

The Use of Induced Pluripotent Stem Cells for the Study and Treatment of Liver Diseases 诱导多能干细胞在肝脏疾病研究和治疗中的应用

Current protocols in toxicology Pub Date : 2016-02-01 DOI: 10.1002/0471140856.tx1413s67

Marc C. Hansel, Julio C. Davila, Massoud Vosough, Roberto Gramignoli, Kristen J. Skvorak, Kenneth Dorko, Fabio Marongiu, William Blake, Stephen C. Strom

{"title":"The Use of Induced Pluripotent Stem Cells for the Study and Treatment of Liver Diseases","authors":"Marc C. Hansel, Julio C. Davila, Massoud Vosough, Roberto Gramignoli, Kristen J. Skvorak, Kenneth Dorko, Fabio Marongiu, William Blake, Stephen C. Strom","doi":"10.1002/0471140856.tx1413s67","DOIUrl":"10.1002/0471140856.tx1413s67","url":null,"abstract":"Liver disease is a major global health concern. Liver cirrhosis is one of the leading causes of death in the world and currently the only therapeutic option for end-stage liver disease (e.g., acute liver failure, cirrhosis, chronic hepatitis, cholestatic diseases, metabolic diseases, and malignant neoplasms) is orthotropic liver transplantation. Transplantation of hepatocytes has been proposed and used as an alternative to whole organ transplant to stabilize and prolong the lives of patients in some clinical cases. Although these experimental therapies have demonstrated promising and beneficial results, their routine use remains a challenge due to the shortage of donor livers available for cell isolation, variable quality of those tissues, the potential need for lifelong immunosuppression in the transplant recipient, and high costs. Therefore, new therapeutic strategies and more reliable clinical treatments are urgently needed. Recent and continuous technological advances in the development of stem cells suggest they may be beneficial in this respect. In this review, we summarize the history of stem cell and induced pluripotent stem cell (iPSC) technology in the context of hepatic differentiation and discuss the potential applications the technology may offer for human liver disease modeling and treatment. This includes developing safer drugs and cell-based therapies to improve the outcomes of patients with currently incurable health illnesses. We also review promising advances in other disease areas to highlight how the stem cell technology could be applied to liver diseases in the future. © 2016 by John Wiley & Sons, Inc.","PeriodicalId":72743,"journal":{"name":"Current protocols in toxicology","volume":"67 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/0471140856.tx1413s67","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50791314","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: 31

A Simple Light Stimulation of Caenorhabditis elegans 秀丽隐杆线虫的简单光刺激

Current protocols in toxicology Pub Date : 2016-02-01 DOI: 10.1002/0471140856.tx1121s67

Kun He Lee, Michael Aschner

引用次数: 6