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Current Protocols in Pharmacology

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https://www.readbyqxmd.com/read/28306153/logical-experimental-design-and-execution-in-the-biomedical-sciences
#1
Daniel J Holder, Michael J Marino
Lack of reproducibility has been highlighted as a significant problem in biomedical research. The present unit is devoted to describing ways to help ensure that research findings can be replicated by others, with a focus on the design and execution of laboratory experiments. Essential components for this include clearly defining the question being asked, using available information or information from pilot studies to aid in the design the experiment, and choosing manipulations under a logical framework based on Mill's "methods of knowing" to build confidence in putative causal links...
March 17, 2017: Current Protocols in Pharmacology
https://www.readbyqxmd.com/read/28306152/assessment-of-ex-vivo-transport-function-in-isolated-rodent-brain-capillaries
#2
Gary N Y Chan, Ronald E Cannon
The blood-brain barrier plays an important role in neuroprotection; however, it can be a major obstacle for drug delivery to the brain. This barrier primarily resides in the brain capillaries and functions as an interface between the brain and peripheral blood circulation. Several anatomical and biochemical elements of the blood-brain barrier are essential to regulate the permeability of nutrients, ions, hormones, toxic metabolites, and xenobiotics into and out of the brain. In particular, high expression of ATP-driven efflux transporters at the blood-brain barrier is a major obstacle in the delivery of CNS pharmacotherapeutics to the brain...
March 17, 2017: Current Protocols in Pharmacology
https://www.readbyqxmd.com/read/28306151/the-quantitative-characterization-of-functional-allosteric-effects
#3
Terry Kenakin
Seven-transmembrane receptors (7TMRs or GPCRs [G protein-coupled receptors]) are nature's prototypic allosteric proteins in that they mediate the interaction between ligand binding to the receptor and the receptor interacting with another cell signaling protein. A growing class of potential drugs acting through 7TMRs are allosteric in nature in that they bind to a separate site on the receptor protein to modify the interactions between the receptor, natural binding/orthosteric ligand, and signaling proteins...
March 17, 2017: Current Protocols in Pharmacology
https://www.readbyqxmd.com/read/28306150/biomarkers-a-general-review
#4
Jeffrey K Aronson, Robin E Ferner
A biomarker is a biological observation that substitutes for and ideally predicts a clinically relevant endpoint or intermediate outcome that is more difficult to observe. The use of clinical biomarkers is easier and less expensive than direct measurement of the final clinical endpoint, and biomarkers are usually measured over a shorter time span. They can be used in disease screening, diagnosis, characterization, and monitoring; as prognostic indicators; for developing individualized therapeutic interventions; for predicting and treating adverse drug reactions; for identifying cell types; and for pharmacodynamic and dose-response studies...
March 17, 2017: Current Protocols in Pharmacology
https://www.readbyqxmd.com/read/27960031/functional-studies-of-sodium-channels-from-target-to-compound-identification
#5
Daniel Bertrand, Bruno Biton, Thomas Licher, Jean-Marie Chambard, Christophe Lanneau, Michel Partiseti, Isabel A Lefevre
Over the last six decades, voltage-gated sodium (Nav ) channels have attracted a great deal of scientific and pharmaceutical interest, driving fundamental advances in both biology and technology. The structure and physiological function of these channels have been extensively studied; clinical and genetic data have uncovered their implication in diseases such as epilepsy, arrhythmias, and pain, bringing them into focus as current and future drug targets. While different techniques have been established to record the activity of Nav channels, proper determination of their properties still presents serious challenges, depending upon the experimental conditions and the desired subtype of channel to be characterized...
December 13, 2016: Current Protocols in Pharmacology
https://www.readbyqxmd.com/read/27960030/detection-of-receptor-heteromerization-using-in-situ-proximity-ligation-assay
#6
Ivone Gomes, Salvador Sierra, Lakshmi A Devi
Although G protein-coupled receptor (GPCR) heteromerization has been extensively demonstrated in vitro using heterologous cells that overexpress epitope-tagged receptors, their presence in endogenous systems is less well established. This is because a criterion to identify receptor heteromerization is the demonstration that the two interacting receptors are present not only in the same cell but also in the same subcellular compartment in close enough proximity to allow for direct receptor-receptor interaction...
December 13, 2016: Current Protocols in Pharmacology
https://www.readbyqxmd.com/read/27960029/stability-of-drugs-drug-candidates-and-metabolites-in-blood-and-plasma
#7
Gregory A Reed
Determination of drug or drug metabolite concentrations in biological samples, particularly in serum or plasma, is fundamental to describing the relationships between administered dose, route of administration, and time after dose for achieving the optimal clinical response. While a well-characterized, accurate analytical method is needed to define these parameters, it must also be established that the analyte concentration in the sample at the time of analysis is identical to the concentration at sample acquisition...
December 13, 2016: Current Protocols in Pharmacology
https://www.readbyqxmd.com/read/27960028/overview-on-research-and-clinical-applications-of-optogenetics
#8
Chris Towne, Kimberly R Thompson
Optogenetics is a method that uses light to control cells in living tissue, typically neurons, that have been modified to express light-sensitive ion channels and pumps. The approach facilitates neuromodulation in brain preparations and freely moving animals with unmatched spatial and temporal resolution. This optogenetics overview describes the vast array of light-sensitive proteins available and the methods used to deliver them to tissue and modulate them with light. How these methods have so far enhanced our knowledge of fundamental neuroscience and psychiatric disease will be discussed as well as how they may contribute to drug discovery in the future...
December 13, 2016: Current Protocols in Pharmacology
https://www.readbyqxmd.com/read/27960027/visualization-of-5-ht-receptors-using-radioligand-binding-autoradiography
#9
Roser Cortés, M Teresa Vilaró, Guadalupe Mengod
Described in this unit are techniques to visualize the majority of serotonin (5-hydroxytryptamine, 5-HT) receptor subtypes in sections of frozen brain tissue using receptor autoradiography. Protocols for brain extraction and sectioning, radioligand exposure, autoradiogram generation, and data quantification are provided, as are the optimal incubation conditions for the autoradiographic visualization of receptors using agonist and antagonist radioligands. © 2016 by John Wiley & Sons, Inc.
December 13, 2016: Current Protocols in Pharmacology
https://www.readbyqxmd.com/read/27636110/a-mouse-model-of-furosemide-induced-overactive-bladder
#10
Michael S Saporito, Eva Zuvich, Amy DiCamillo
Detailed in this unit is a mouse model of overactive bladder and urinary incontinence based on diuretic stress-induced urination. The procedure involves the use of a unique, highly sensitive, and automated urine capturing method to measure urinary latency, frequency, and void volume. Although this method was first described and validated using an anti-muscarinic drug used for treating overactive bladder, subsequent work has shown that effective non-cholinergic agents can be detected. These findings indicate good predictive value for this model regarding the possible clinical utility of test agents as treatments for overactive bladder, regardless of their site of action...
September 16, 2016: Current Protocols in Pharmacology
https://www.readbyqxmd.com/read/27248578/genetically-engineered-mouse-models-of-pancreatic-cancer-the-kpc-model-lsl-kras-g12d-lsl-trp53-r172h-pdx-1-cre-its-variants-and-their-application-in-immuno-oncology-drug-discovery
#11
Jae W Lee, Chad A Komar, Fee Bengsch, Kathleen Graham, Gregory L Beatty
Pancreatic ductal adenocarcinoma (PDAC) ranks fourth among cancer-related deaths in the United States. For patients with unresectable disease, treatment options are limited and lack curative potential. Preclinical mouse models of PDAC that recapitulate the biology of human pancreatic cancer offer an opportunity for the rational development of novel treatment approaches that may improve patient outcomes. With the recent success of immunotherapy for subsets of patients with solid malignancies, interest is mounting in the possible use of immunotherapy for the treatment of PDAC...
June 1, 2016: Current Protocols in Pharmacology
https://www.readbyqxmd.com/read/27248577/mammalian-cell-tissue-culture-techniques
#12
Katy Phelan, Kristin M May
Cultured tissues and cells are used extensively in physiological and pharmacological studies. In vitro cultures provide a means of examining cells and tissues without the complex interactions that would be present if the whole organism were studied. A number of special skills are required in order to preserve the structure, function, behavior, and biology of cells in culture. This unit describes the basic skills required to maintain and preserve cell cultures: maintaining aseptic technique, preparing media with the appropriate characteristics, passaging, freezing and storage, recovering frozen stocks, and counting viable cells...
June 1, 2016: Current Protocols in Pharmacology
https://www.readbyqxmd.com/read/26995550/drug-combinations-tests-and-analysis-with-isoboles
#13
Ronald J Tallarida
Described in this unit are experimental and computational methods to detect and classify drug interactions. In most cases, this relates to two drugs or compounds with overtly similar effects, e.g., two analgesics or two anti-hypertensives. From the dose-response data of the individual drugs, it is possible to generate a curve, the isobole, which defines all dose combinations that are expected to yield a specified effect. The theory underlying the isobole involves the calculation of doses of drug A that are effectively equivalent to doses of drug B with that equivalence determining whether the isobole is linear or nonlinear...
March 18, 2016: Current Protocols in Pharmacology
https://www.readbyqxmd.com/read/26995549/models-of-inflammation-carrageenan-air-pouch
#14
Djane B Duarte, Michael R Vasko, Jill C Fehrenbacher
The subcutaneous air pouch is an in vivo model that can be used to study the components of acute and chronic inflammation, the resolution of the inflammatory response, the oxidative stress response, and potential therapeutic targets for treating inflammation. 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...
March 18, 2016: Current Protocols in Pharmacology
https://www.readbyqxmd.com/read/26995548/methods-of-inducing-inflammatory-bowel-disease-in-mice
#15
Byoungwook Bang, Lenard M Lichtenberger
Animal models of experimentally induced inflammatory bowel disease (IBD) are useful for understanding more about the mechanistic basis of the disease, identifying new targets for therapeutic intervention, and testing novel therapeutics. This unit provides detailed protocols for five widely used mouse models of experimentally induced intestinal inflammation: chemical induction of colitis by dextran sodium sulfate (DSS), hapten-induced colitis via 2,4,6-trinitrobenzene sulfonic acid (TNBS), Helicobacter-induced colitis in mdr1a(-/-) mice, the CD4(+) CD45RB(hi) SCID transfer colitis model, and the IL-10(-/-) colitis model...
March 18, 2016: Current Protocols in Pharmacology
https://www.readbyqxmd.com/read/26995547/overview-of-genetically-engineered-mouse-models-of-distinct-breast-cancer-subtypes
#16
REVIEW
Jerry Usary, David Brian Darr, Adam D Pfefferle, Charles M Perou
Advances in the screening of new therapeutic options have significantly reduced the breast cancer death rate over the last decade. Despite these advances, breast cancer remains the second leading cause of cancer death among women. This is due in part to the complexity of the disease, which is characterized by multiple subtypes that are driven by different genetic mechanisms and that likely arise from different cell types of origin. Because these differences often drive treatment options and outcomes, it is important to select relevant preclinical model systems to study new therapeutic interventions and tumor biology...
March 18, 2016: Current Protocols in Pharmacology
https://www.readbyqxmd.com/read/26995546/overview-of-transgenic-glioblastoma-and-oligoastrocytoma-cns-models-and-their-utility-in-drug-discovery
#17
REVIEW
Fuyi Chen, Albert Becker, Joseph LoTurco
Many animal models have been developed to investigate the sources of central nervous system (CNS) tumor heterogeneity. Reviewed in this unit is a recently developed CNS tumor model using the piggyBac transposon system delivered by in utero electroporation, in which sources of tumor heterogeneity can be conveniently studied. Their applications for studying CNS tumors and drug discovery are also reviewed. © 2016 by John Wiley & Sons, Inc.
March 18, 2016: Current Protocols in Pharmacology
https://www.readbyqxmd.com/read/27636112/overview-of-critical-parameters-for-the-design-and-execution-of-a-high-throughput-screen-for-allosteric-ligands
#18
Andrew Alt
Allosteric ligands modulate the activity of receptor targets by binding to sites that are distinct from the orthosteric (native agonist) binding site. Allosteric modulators have potential therapeutic advantages over orthosteric agonists and antagonists, including improved selectivity, and maintenance of the spatial and temporal fidelity of native signaling patterns. The identification of allosteric ligands presents unique challenges because of the requirement for screening in the presence of an orthosteric agonist, the small signal window that is produced by many allosteric modulators, the proclivity of allosteric modulators to exhibit activity switching within a chemotype (e...
2016: Current Protocols in Pharmacology
https://www.readbyqxmd.com/read/27636111/in-vitro-drug-metabolism-using-liver-microsomes
#19
Kathleen M Knights, David M Stresser, John O Miners, Charles L Crespi
Knowledge of the metabolic stability of newly discovered drug candidates eliminated by metabolism is essential for predicting the pharmacokinetic (PK) parameters that underpin dosing and dosage frequency. Further, characterization of the enzyme(s) responsible for metabolism (reaction phenotyping) allows prediction, at least at the qualitative level, of factors (including metabolic drug-drug interactions) likely to alter the clearance of both new chemical entities (NCEs) and established drugs. Microsomes are typically used as the enzyme source for the measurement of metabolic stability and for reaction phenotyping because they express the major drug-metabolizing enzymes cytochrome P450 (CYP) and UDP-glucuronosyltransferase (UGT), along with others that contribute to drug metabolism...
2016: Current Protocols in Pharmacology
https://www.readbyqxmd.com/read/27636109/measurement-of-receptor-signaling-bias
#20
Terry Kenakin
G protein-coupled receptors (GPCRs) are often pleiotropically linked to numerous cellular signaling mechanisms in cells, and it is now known that many agonists differentially activate some signaling pathways at the expense of others. The mechanism for this effect is the stabilization of different active receptor states by different agonists, and it leads to varying qualities of efficacy for different agonists. Agonist bias is a powerful mechanism to amplify beneficial signals and diminish harmful signals, and thus improve the overall profile of agonist ligands...
2016: Current Protocols in Pharmacology
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