Preclinical orthotopic and intracardiac injection models of human breast cancer metastasis to bone and their use in drug discovery.
Authors: Scepansky E, Goldstein R, Rosenblatt M
Abstract
Breast cancer is the most common malignancy in women, with the development of distant metastases rendering the condition incurable. Relatively little is known about the factors governing the progression from primary tumor to metastasis, in part because of the difficulty in modeling what is a complex series of events. Detailed in this unit are descriptions of two murine models of breast cancer metastasis to bone that can be used to screen the effectiveness of new chemical entities on this disease process.
PMID: 21935904 [PubMed - indexed for M...
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
Measurement of action potential generation in isolated canine left ventricular midmyocardial myocytes.
Authors: Johnson DM, Hussein L, Spätjens RL, Valentin JP, Volders PG, Abi-Gerges N
Abstract
Proarrhythmic side effects are a major limitation during the drug development process for cardiac and non-cardiac compounds. Because changes in cardiac action potential (AP) are undesirable, the evaluation of the effects of test compounds on the AP is essential before advancing new compounds to clinical testing. However, an increase in repolarization duration alone is not always proarrhythmic, and newer surrogate markers have been suggested to better predict the occurrence of arrhythmia. Described in this unit is a...
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
Measurement of glucose uptake in cultured cells.
Authors: Yamamoto N, Ueda M, Sato T, Kawasaki K, Sawada K, Kawabata K, Ashida H
Abstract
Facilitative glucose uptake transport systems are ubiquitous in animal cells and responsible for transporting glucose across the cell surface membrane. Evaluation of glucose uptake is crucial in the study of numerous diseases and metabolic disorders, such as myocardial ischemia, diabetes mellitus, and cancer. Methods for assessing glucose uptake into mammalian cells are detailed in this unit. The work is divided into four sections: (1) a brief overview of glucose uptake assays in cultured cells; (2) a method for measur...
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
In vitro opioid receptor assays.
Authors: Taylor DA
Abstract
Although opioid analgesics have been used for centuries, identification of opioid receptors and the ability of an opioid receptor antagonist to block natural pain processes prompted a search for endogenous opioid peptides. In vitro models were needed to characterize opioid activity in biological samples. The longitudinal muscle/myenteric plexus (LM/MP) of the guinea pig ileum was the classical in vitro assay system, but the development of the mouse vas deferens (MVD) assay provided another important model that could be employed. Both assays entail electrical stimulation of intra...
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
Overview of drug product development.
Authors: Narayan P
Abstract
The process for developing drug delivery systems has evolved over the past two decades with more scientific rigor, involving a collaboration of various fields, i.e., biology, chemistry, engineering, and pharmaceutics. Drug products, also commonly known in the pharmaceutical industry as formulations or "dosage forms," are used for administering the active pharmaceutical ingredient (API) for purposes of assessing safety in preclinical models, early- to late-phase human clinical trials, and for routine clinical/commercial use. This overview discusses approaches for creating small-m...
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
Models of neurological disease (substance abuse): self-administration in monkeys.
Authors: Platt DM, Carey G, Spealman RD
Abstract
Drug self-administration is a procedure in which a subject performs a specified response that results in the delivery of a drug injection. This procedure is viewed as a relevant model for the study of human drug-taking behavior. Drug self-administration in primates has several characteristics that resemble drug-taking behavior in humans, and agents commonly abused by humans also generally maintain self-administration behavior in monkeys. Self-administration procedures allow for the study of a variety of drug properties. For instance, they can be used to inve...
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
Measurement of cell death in Mammalian cells.
Authors: Cummings BS, Wills LP, Schnellmann RG
Abstract
Methods for assessing mammalian cell death are presented in this unit. The unit is divided into six sections: (1) a brief overview of cytotoxicity and pathways of cell death, (2) a method to measure cell death using lactate dehydrogenase (LDH) release as a marker of membrane integrity, (3) a flow cytometry method that simultaneously measures two types of cell death, necrosis, and apoptosis, (4) use of fluorescence microscopy and nuclear morphology to assess apoptosis and necrosis, (5) the use of multi-well plates and high-content analysis imaging syst...
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
Evaluating modulators of "Regulator of G-protein Signaling" (RGS) proteins.
Authors: Bosch DE, Zielinski T, Lowery RG, Siderovski DP
Abstract
"Regulator of G-protein Signaling" (RGS) proteins constitute a class of intracellular signaling regulators that accelerate GTP hydrolysis by heterotrimeric Gα subunits. In recent years, RGS proteins have emerged as potential drug targets for modulation by small molecules. Described in this unit are high-throughput screening procedures for identifying modulators of RGS protein-mediated GTPase acceleration (GAP activity), for assessment of RGS domain/Gα interactions (most avid in vitro when Gα is bound by aluminum tetrafluoride), and for va...
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
Models of inflammation: Carrageenan- or complete Freund's Adjuvant (CFA)-induced edema and hypersensitivity in the rat.
Authors: Fehrenbacher JC, Vasko MR, Duarte DB
Abstract
Animal models of inflammation are used to assess the production of inflammatory mediators at sites of inflammation, the anti-inflammatory properties of agents such as nonsteroidal anti-inflammatory drugs (NSAIDs), and the efficacy of putative analgesic compounds in reversing cutaneous hypersensitivity. This unit details methods to elicit and measure carrageenan- and complete Freund's adjuvant (CFA)-induced cutaneous inflammation. Due to possible differences between the dorsal root sensory system and the trigeminal sensory system, injections of either t...
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
Models of inflammation: carrageenan air pouch.
Authors: Duarte DB, Vasko MR, Fehrenbacher JC
Abstract
The subcutaneous air pouch is an in vivo model that can be used to study acute and chronic inflammation, the resolution of the inflammatory response, and the oxidative stress response. Injection of irritants into an air pouch in rats or mice induces an inflammatory response that can be quantified by the volume of exudate produced, the infiltration of cells, and the release of inflammatory mediators. The model presented in this unit has been extensively used to identify potential anti-inflammatory drugs.
PMID: 22383000 [PubMed - indexed for MEDL...
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
Whole-cell configuration of the patch-clamp technique in the hERG channel assay to predict the ability of a compound to prolong QT interval.
Authors: Goineau S, Legrand C, Froget G
Abstract
In vitro electrophysiological safety studies have become an integral part of the drug development process since, in many instances, compound-induced QT prolongation has been associated with a direct block of human ether-a-go-go-related gene (hERG) potassium channels or its native current, the rapidly activating delayed rectifier potassium current (I(Kr)). Therefore, the in vitro hERG channel patch-clamp assay is commonly used as an early screen to predict the ability of a compound to prolong QT interval. The protocol described in this unit is designed to ass...
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
Topoisomerase assays.
Authors: Nitiss JL, Soans E, Rogojina A, Seth A, Mishina M
Abstract
Topoisomerases are nuclear enzymes that play essential roles in DNA replication, transcription, chromosome segregation, and recombination. All cells have two major forms of topoisomerases: type I enzymes, which make single-stranded cuts in DNA, and type II enzymes, which cut and pass double-stranded DNA. DNA topoisomerases are important targets of approved and experimental anti-cancer agents. The protocols described in this unit are for assays used to assess new chemical entities for their ability to inhibit both forms of DNA topoisomerase...
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
Models of affective illness: chronic mild stress in the rat.
Authors: Papp M
Abstract
Described in this unit is a chronic mild stress (CMS) procedure used for predicting an antidepressant response. Following exposure to a variety of mild stressors for a period of several weeks, rat behavior is modified in a number of ways. Among these is a substantial reduction in consumption of a 1% sucrose solution. Chronic administration of antidepressant drugs reverses diminished enthusiasm for sucrose in these subjects. While most antidepressants must be administered for at least 3 to 5 weeks to normalize behavior, there are treatments that display a more rapid onset of action....
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
High throughput functional assays for P2X receptors.
Authors: Namovic MT, Jarvis MF, Donnelly-Roberts D
Abstract
Adenosine triphosphate (ATP) activates two receptor superfamilies, metabotropic P2Y and ionotropic P2X receptors. The P2X receptors are nonselective cation channels that are widely expressed on excitable cells including neurons, glia, and smooth muscle cells. The protocols in this unit are useful for evaluating ligands that interact with P2X receptors on native cells or that are cloned and expressed in recombinant heterologous cell systems. Calcium imaging methods are described for the pharmacological characterization of fast or slowly desensitizi...
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
Overview of animal models of obesity.
Authors: Lutz TA, Woods SC
Abstract
The focus of this overview is on the animal models of obesity most commonly utilized in research. The models include monogenic models in the leptin pathway, polygenic diet-dependent models, and, in particular for their historical perspective, surgical and chemical models of obesity. However, there are far too many models to consider all of them comprehensively, especially those caused by selective molecular genetic approaches modifying one or more genes in specific populations of cells. Further, the generation and use of inducible transgenic animals (induced knock-out or...
Source: Current Protocols in Pharmacology - November 18, 2015 Category: Drugs & Pharmacology Tags: Curr Protoc Pharmacol Source Type: research
