Current Protocols in Pharmacology最新文献_第4页

Modeling Chronic Graft Versus Host Disease in Mice Using Allogeneic Bone Marrow and Splenocyte Transfer. 用同种异体骨髓和脾细胞移植模拟小鼠慢性移植物抗宿主病。

Current Protocols in Pharmacology Pub Date : 2018-12-01 Epub Date: 2018-09-11 DOI: 10.1002/cpph.47

Mark A Schroeder, Kidist Ashami, Karl Staser

{"title":"Modeling Chronic Graft Versus Host Disease in Mice Using Allogeneic Bone Marrow and Splenocyte Transfer.","authors":"Mark A Schroeder, Kidist Ashami, Karl Staser","doi":"10.1002/cpph.47","DOIUrl":"https://doi.org/10.1002/cpph.47","url":null,"abstract":"This unit describes a method for allogeneic bone marrow and splenocyte transfer for the modeling of chronic graft versus host disease (cGVHD) in mice. Preclinical models provide clinically relevant platforms for mechanistic and therapeutic studies that may inform the treatment of patients suffering from cGVHD, a common and potentially severe complication of allogeneic hematopoietic stem cell transplantation (alloHSCT). Most murine models of cGVHD depend on the transfer of major histocompatibility complex (MHC)-mismatched bone marrow and whole splenocytes (or purified T cells) into an irradiated recipient. The bone marrow contains hematopoietic stem and progenitor cells necessary to reconstitute the irradiated host hematopoietic system, while splenocytes contain T cells that mediate cGVHD. Of note, specific mouse strains, splenocyte dose, bone marrow quantity, and irradiation doses vary widely across different cGVHD models. Here we describe donor bone marrow and splenocyte preparation, recipient irradiation and intravenous injection of donor cells, and clinical monitoring for disease emergence and progression. © 2018 by John Wiley & Sons, Inc.","PeriodicalId":10871,"journal":{"name":"Current Protocols in Pharmacology","volume":"83 1","pages":"e47"},"PeriodicalIF":0.0,"publicationDate":"2018-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpph.47","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36481801","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 4

Issue Information TOC 发布信息TOC

Current Protocols in Pharmacology Pub Date : 2018-11-21 DOI: 10.1002/cpph.49

引用次数: 0

Overview of Transporters in Pharmacokinetics and Drug Discovery 转运蛋白在药代动力学和药物发现中的概述

Current Protocols in Pharmacology Pub Date : 2018-08-31 DOI: 10.1002/cpph.46

Yan Zhang

引用次数: 6

Screening Strategies for the Discovery of Ion Channel Monoclonal Antibodies 发现离子通道单克隆抗体的筛选策略

Current Protocols in Pharmacology Pub Date : 2018-08-31 DOI: 10.1002/cpph.44

Caroline S. Colley, Elizabeth England, John E. Linley, Trevor C. I. Wilkinson

{"title":"Screening Strategies for the Discovery of Ion Channel Monoclonal Antibodies","authors":"Caroline S. Colley, Elizabeth England, John E. Linley, Trevor C. I. Wilkinson","doi":"10.1002/cpph.44","DOIUrl":"10.1002/cpph.44","url":null,"abstract":"Ion channels play crucial roles in physiology by modulation of cellular functions that include electrical excitability, secretion, cell migration, and gene transcription. They are an important target class for drug discovery and have historically been targeted using small molecule approaches. A significant opportunity exists to target these channels with antibodies and alternative forms of biologics. Antibodies display high specificity, selectivity, and affinity for their target antigen, thus having the potential to target ion channels very precisely. Nonetheless, isolating antibodies to ion channels is challenging due to the difficulties in expression and purification of ion channels in a format suitable for antibody drug discovery and due to the complexities of screening for function. In this overview, we focus on an array of screening methods, ranging from direct antibody binding screens to complex electrophysiological assays, and describe how these assays can be used to identify functional monoclonal antibodies. We also provide some insights into specific considerations which are required to enable these screens to be used for antibody drug discovery. © 2018 by John Wiley & Sons, Inc.","PeriodicalId":10871,"journal":{"name":"Current Protocols in Pharmacology","volume":"82 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpph.44","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36448457","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: 6

Protocols for Measuring Glutamate Uptake: Dose-Response and Kinetic Assays in In Vitro and Ex Vivo Systems 谷氨酸摄取测定方案:体外和离体系统的剂量-反应和动力学分析

Current Protocols in Pharmacology Pub Date : 2018-08-27 DOI: 10.1002/cpph.45

Andréia C. K. Fontana

{"title":"Protocols for Measuring Glutamate Uptake: Dose-Response and Kinetic Assays in In Vitro and Ex Vivo Systems","authors":"Andréia C. K. Fontana","doi":"10.1002/cpph.45","DOIUrl":"10.1002/cpph.45","url":null,"abstract":"This article presents detailed descriptions of procedures and troubleshooting tips for basic in vitro and ex vivo uptake assays for the functional characterization of glutamate transporters and the assessment of the effect of compounds that modulate their activity. Assays are performed in cell lines that transiently or stably express a particular transporter under investigation, in primary cultures of astrocytes, or in ex vivo synaptosomal preparations that endogenously express these transporters. Two main assays are described, including dose-response assays to measure potencies of test compounds for stimulation or inhibition of function (EC50 or IC50 values, respectively) and kinetic functional assays to calculate apparent affinity (KM) and maximal velocity (Vmax) of radiolabeled substrate uptake into the cells. The methods described use glutamate transporters as an example; however, the protocols can be adapted to other neurotransmitter transporters using their respective substrates (e.g., GABA uptake through GATs, serotonin uptake through SERTs, among others). © 2018 by John Wiley & Sons, Inc.","PeriodicalId":10871,"journal":{"name":"Current Protocols in Pharmacology","volume":"82 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpph.45","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36431816","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: 5

Utilizing Miniature Fluorescence Microscopy to Image Hippocampal Place Cell Ensemble Function in Thy1.GCaMP6f Transgenic Mice 利用微型荧光显微镜成像海马位置细胞集合功能。GCaMP6f转基因小鼠

Current Protocols in Pharmacology Pub Date : 2018-08-20 DOI: 10.1002/cpph.42

Tim Indersmitten, Tamara Berdyyeva, Leah Aluisio, Timothy Lovenberg, Pascal Bonaventure, Ryan M. Wyatt

引用次数: 3

Overview of Genetically Engineered Mouse Models of Prostate Cancer and Their Applications in Drug Discovery 前列腺癌基因工程小鼠模型及其在药物开发中的应用综述

Current Protocols in Pharmacology Pub Date : 2018-06-07 DOI: 10.1002/cpph.39

Gianluca Civenni, Giuseppina M. Carbone, Carlo V. Catapano

{"title":"Overview of Genetically Engineered Mouse Models of Prostate Cancer and Their Applications in Drug Discovery","authors":"Gianluca Civenni, Giuseppina M. Carbone, Carlo V. Catapano","doi":"10.1002/cpph.39","DOIUrl":"10.1002/cpph.39","url":null,"abstract":"Prostate cancer (PCa) is the most common malignant visceral neoplasm in males in Western countries. Despite progress made in the early treatment of localized malignancies, there remains a need for therapies effective against advanced forms of the disease. Genetically engineered mouse (GEM) models are valuable tools for addressing this issue, particularly in defining the cellular and molecular mechanisms responsible for tumor initiation and progression. While cell and tissue culture systems are important models for this purpose as well, they cannot recapitulate the complex interactions within heterotypic cells and the tumor microenvironment that are crucial in the initiation and progression of prostate tumors. Limitations of GEM models include resistance to developing invasive and metastatic tumors that resemble the advanced stages of human PCa. Nonetheless, because genetic models provide valuable information on the human condition that would otherwise be impossible to obtain, they are increasingly employed to identify molecular targets and to examine the efficacy of cancer therapeutics. The aim of this overview is to provide a brief but comprehensive summary of GEM models for PCa, with particular emphasis on the strengths and weaknesses of this experimental approach. © 2018 by John Wiley & Sons, Inc.","PeriodicalId":10871,"journal":{"name":"Current Protocols in Pharmacology","volume":"81 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-06-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpph.39","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36243846","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: 7

Measuring Histone Deacetylase Inhibition in the Brain 测定脑组织组蛋白去乙酰化酶抑制作用

Current Protocols in Pharmacology Pub Date : 2018-06-07 DOI: 10.1002/cpph.41

Doodipala Samba Reddy, Xin Wu, Victoria M. Golub, W. Mohaiza Dashwood, Roderick H. Dashwood

引用次数: 27

Reagent Validation to Facilitate Experimental Reproducibility 试剂验证以促进实验可重复性

Current Protocols in Pharmacology Pub Date : 2018-05-16 DOI: 10.1002/cpph.40

Michael Williams

{"title":"Reagent Validation to Facilitate Experimental Reproducibility","authors":"Michael Williams","doi":"10.1002/cpph.40","DOIUrl":"10.1002/cpph.40","url":null,"abstract":"Many results reported in the biomedical research literature cannot be independently reproduced, undermining the basic foundations of science. This overview is intended for researchers who are committed to improving the quality and integrity of biomedical science by raising awareness of both the sources of irreproducibility, and activities specifically targeted to address the issue. The irreproducibility of biomedical research is due to a variety of factors, known and unknown, that markedly influence experimental outcomes. Among the known factors are inadequate training or mentoring, or experimenter incompetence. These may result in flawed experimental design and execution that reflect a lack of planning, leading to an underpowering of a study, an absence of appropriate controls, selective data analysis, inappropriate statistical analysis, and/or investigator bias or fraud. Another frequently overlooked source of irreproducibility is failure to ensure that the equipment used is performing up to manufacturer specifications. Failure to validate/authenticate the experimental reagents is another source of irreproducibility. These include compounds, cell lines, antibodies, and the animal models used in a study. This overview discusses how a lack of validation of research reagents negatively impact experimental reproducibility, and provides guidelines to avoid these pitfalls. © 2018 by John Wiley & Sons, Inc.","PeriodicalId":10871,"journal":{"name":"Current Protocols in Pharmacology","volume":"81 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpph.40","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36243847","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: 7

Reconstitution and Electrophysiological Characterization of Ion Channels in Lipid Bilayers 脂质双分子层离子通道的重构与电生理表征

Current Protocols in Pharmacology Pub Date : 2018-05-04 DOI: 10.1002/cpph.37

Dan A. Klaerke, Maria de los Angeles Tejada, Vibeke Grøsfjeld Christensen, Mette Lassen, Per Amstrup Pedersen, Kirstine Calloe

引用次数: 6