bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cysteinyl Leukotriene Receptor Assays
Authors: Muccitelli Roseanna M. , Luttmann Mark A. , Hay Douglas W.P. 
Published: June 2004

AbstractThe cysteinyl leukotrienes (CysLTs), LTC4, LTD4 and LTE4, lipid products derived from arachidonic acid metabolism, have been implicated in the pathophysiology of several inflammatory diseases, in particular, asthma. This unit describes techniques and applications for the measurement of contractile responses to the CysLTs in isolated smooth muscle preparations. The contractions are assessed by standard methods for the isometric measurement of responses (contractile or relaxant) of isolated tissues to exogenous agonists, and a detailed description of the methods employed to assess CysLT‐induced contractions in guinea‐pig trachea is outlined. However, the same general methodology (other than parameters such as dissection for non‐airway tissues) are appropriate for measuring CysLT‐induced contractions in airway preparations from other animals, and in non‐airways tissues (e.g., the gastrointestinal tract) from different species, and also in exploring the relaxant responses to the CysLTs that have been demonstrated in some tissues (e.g., pulmonary vein or artery).
Models of Inflammation: Carrageenan‐Induced Paw Edema in the Rat
Authors: Whiteley Phyllis E. , Dalrymple Stacie A. 
Published: June 2004

AbstractPaw swelling, or footpad edema, is a convenient method for assessing inflammatory responses to antigenic challenges and irritants. The protocol described in this unit uses carrageenan as the irritant to induce paw edema. Typically, test compounds are assessed for acute anti‐inflammatory activity by examining their ability to reduce or prevent the development of carrageenan‐induced paw swelling. This model has long been used to assess the anti‐inflammatory properties of agents such as nonsteroidal anti‐inflammatory drugs (NSAIDs) that inhibit prostaglandin production.
Models of Inflammation: Adjuvant‐Induced Arthritis in the Rat
Authors: Whiteley Phyllis E. , Dalrymple Stacie A. 
Published: June 2004

AbstractInjection of adjuvant (Mycobacterium butyricum suspended in mineral oil) into rats produces an immune reaction that characteristically involves inflammatory destruction of cartilage and bone of the distal joints with concomitant swelling of surrounding tissues. Adjuvant‐induced arthritis in rats is commonly used to evaluate compounds that might be of potential use as drugs for treatment of rheumatoid arthritis and other chronic inflammatory conditions. This unit describes a method for inducing arthritis by injecting adjuvant into the tail and evaluating a test compound for the ability to inhibit the inflammatory response.
Models of Urogenital Dysfunction: Urinary Urge Incontinence (UUI) in Anesthetized Dog and Rat Models
Authors: Newgreen Donald , Anderson Charles , Nunn Philip , Carter Anthony , Bushfield Mark 
Published: June 2004

AbstractThis unit describes a model system for the measurement of the effects of compounds on bladder function in anesthetized dog and rat. A protocol is also described for measuring nerve‐stimulated salivary secretion which is particularly relevant when investigating agents that modulate parasympathetic drive (such as muscarinic antagonists), since it allows for the measurement of relative selectivity of bladder function over salivation. This unit describes a model system for the measurement of the effects of compounds on bladder function in anesthetized dog and
Models of Neurological Disease (Alzheimer's Disease): Spatial Discrimination Water Maze Test in Septal‐Lesioned Rats
Authors: Decker Michael W. 
Published: June 2004

AbstractModels of Alzheimer s disease currently focus on symptomatology, as understanding of the etiology of the disorder is not yet sufficiently advanced to model the disease process in animals. Moreover, currently available treatments are purely palliative in nature. Given that cognitive dysfunction is a central feature of the disease, animal models of cognitive dysfunction are frequently used to evaluate the therapeutic potential of test compounds. This unit presents a rat model in which a brain lesion is produced which disrupts septohippocampal function, thus impairing performance on a spatial learning task (the Morris water maze).
Models of Schizophrenia: Phencyclidine Disruption of Prepulse Inhibition (PPI) of Startle in Rats
Authors: Kehne John H. , Hoffman Diane C. , Cassella James V. 
Published: June 2004

AbstractModern theories of schizophrenia implicate deficits in glutamatergic transmission in the etiology of this disorder. This unit describes an animal model in which a deficit in glutamatergic transmission is induced pharmacologically by the NMDA receptor ion channel blocker phencyclidine. Reversal of the resulting behavioral impairment, the reduction of sensory‐motor gating as measured by prepulse inhibition, can be used as a method for evaluating novel treatments for schizophrenia.
Models for Assessing Antipsychotics: Antagonism of Amphetamine‐Induced Hyperactivity and Stereotypies in Mice
Authors: Roux Sylvain , Hay Anne‐Marie , Porsolt Ruger D. 
Published: June 2004

AbstractAll classical antipsychotics, such as chlorpromazine and haloperidol, have potent dopamine receptor‐blocking propertiesThis unit describes the rat anti‐Thy‐1.1 model of acute proliferative glomerulonephritis for the study of chronic renal insufficiency. A procedure is detailed for the induction of glomerulonephritis in rats as well as measurement of daily urinary excretion of protein, which is a convenient, primary screening tool. The unit also provides methods for assessment of glomerular filtration rate and effective renal plasma flow in anesthetized rats with anti‐Thy‐1.1‐induced renal insufficiency.
Characterization of Matrix Metalloproteinase Inhibitors: Angiogenesis and Tumor Models
Authors: Albert Daniel H. , Davidsen Steven K. 
Published: June 2004

AbstractSince the matrix metalloproteinases (MMPs) have an essential role in the process of tumor growth, invasion and metastasis, small molecule MMP inhibitors have the ability to modulate tumor progression in animals and the potential to be of therapeutic benefit to cancer patients. The antiangiogenic properties of MMP inhibitors can be assessed by the measurement of hemoglobin content of Matrigel plugs containing angiogenic growth factors introduced into the flanks of mice. A flank tumor growth model using B16 murine melanoma cells provides a useful means of determining the antitumor effects of MMP inhibitors as well as correlating efficacy with the concentration of drug in blood.
Overview of Animal Models of Asthma
Authors: Canning Brendan J. 
Published: June 2004

AbstractAsthma is an inflammatory disease characterized by airways obstruction, airways hyperresponsiveness, excessive mucous secretion and cough. Guinea pig airways display many anatomical, physiological and pharmacological attributes of human airways, making this species ideal for modeling the asthmatic condition. This unit provides an overview of animal models of asthma, including definitions, descriptions of available animal models, and discussion of numerous critical issues to consider before designing a model to study this complex disease.
Models of Neuropathic Pain in the Rat
Authors: Bennett Gary J. , Chung Jin Mo , Honore (Table 5.32.2) Marie , Seltzer Ze'ev 
Published: June 2004

AbstractPeripheral nerve injury due to trauma, disease, and certain toxins sometimes produces abnormal (neuropathic) pain syndromes that are chronic and refractory to standard analgesics. Knowledge of the mechanisms that produce neuropathic pain and the ability to search for new drugs to control it have been greatly advanced by the introduction of rat models of post‐traumatic painful peripheral neuropathy. There are currently three models of neuropathic pain in the rat that are widely used. The procedures to create these models and the behavioral assays used to quantify the resulting neuropathic pain symptoms are described in this unit: the chronic constriction injury (CCI) model, the partial sciatic ligation (PSL) model, and the spinal nerve ligation (SNL) model. Four kinds of abnormal pain sensations are commonly measured to assess the outcome: heat‐hyperalgesia, mechano‐hyperalgesia, mechano‐allodynia, and cold‐allodynia.
Electrical Amygdala Kindling
Authors: Dürmüller N. , Porsolt R.D. 
Published: June 2004

AbstractThis unit describes a method of electrical amygdala kindling in the rat. This procedure requires mastery of stereotaxic electrode implantation which is not covered in the current unit. Also, the investigator must have a sound knowledge of electronics and computing. The text gives instructions on how to render rats epileptic, how to determine the effects of compounds in kindled rats, and how to analyze the data. Results with three reference substances are illustrated. These substances are used in the clinic and give robust results in kindling.
A Rat Model of Postoperative Pain
Authors: Brennan Timothy J. 
Published: June 2004

AbstractResearch on pain resulting from an incision is crucial for developing therapies for postoperative pain, a condition for which effective and inexpensive treatments are not yet available. Because of the gap between preclinical models of persistent pain (e.g., inflammatory, neuropathic) and acute postsurgical pain, efforts have been undertaken to develop methods for studying pain caused by incisions. To this end, a rat model for postoperative pain has been developed. The model uses a plantar incision in the hindpaw and is characterized by persistent, reduced withdrawal thresholds to mechanical stimuli, as is the case in patients after surgery. The pain behaviors are greatest immediately after recovery from anesthesia. The enhanced responsiveness remains remarkable for several days and then gradually decreases. The model is useful for screening test compounds that may be effective in reducing postoperative pain and for understanding pain mechanisms associated with incisions.
Isotopic Assays for Reporter Gene Activity
Authors: Kingston Robert E. , Sheen Jen , Moore David 
Published: June 2004

AbstractThis unit describes two widely used reporter systems that are based on radioactive detection assays. The first assay uses chloramphenicol acetyltransferase (CAT) activity as a measure of the level of expression of a transfected gene. This bacterial enzyme catalyzes the transfer of an acyl group from acetyl CoA to chloramphenicol. Transfected cells are harvested and lysed, and then acyl CoA and radioactively labeled chloramphenicol are added to cell lysate, and modified derivatives of the antibiotic are separated from the starting material. The second reporter system uses a kit to perform a simple two‐site radioimmunoassay to quantitate the amount of human growth hormone (hGH) secreted into culture medium by transfected cells. Medium is incubated with 125I‐labeled antibody specific for hGH, and immune complexes are collected by an avidin‐coated bead. The quantity of hormone is determined based on comparison with a standard curve.
Use of the A. Victoria Green Fluorescent Protein to Study Protein Dynamics In Vivo
Authors: Kahana Jason A. , Silver Pam A. 
Published: June 2004

AbstractFluorescent molecules serve as valuable tools for the detection of a variety of biochemical phenomena. Such reagents have been employed for protein localization, quantitation of gene expression, detection of nucleic acids, cell sorting, and determination of chemical concentrations. Although fluorescence is a useful tool for detecting molecules within cells, its application in vivo has been limited. The ideal vital fluorescent tag should (1) be detectable without causing cytological damage, (2) be able to label a wide variety of cell types readily, and (3) be able to be targeted to virtually any subcellular region. The recently cloned green fluorescent protein (GFP) from the jellyfish Aequorea victoria is such a molecule. This overview describes the use of this proteinaceous fluorophore for in vivo observation of cellular phenomena, including applications and problems with the use of GFP, a discussion of mutant GFPs with altered fluorescence characteristics, and also some details on microscopy requirements.
Enzymatic Amplification of DNA by PCR: Standard Procedures and Optimization
Authors: Kramer Martha F. , Coen Donald M. 
Published: June 2004

AbstractThis unit describes a method for amplifying DNA enzymatically by the polymerase chain reaction (PCR), including procedures to quickly determine conditions for successful amplification of the sequence and primer sets of interest, and to optimize for specificity, sensitivity, and yield. Hot‐start methods are described which can greatly improve specificity, sensitivity, and yield. This protocol suggests some relatively inexpensive methods to achieve hot start, and lists several commercial hot‐start options which may be more convenient, but of course more expensive. The unit has recently been updated to include new information on reagents to enhance the reaction, better cycling parameters, and innovations in robotics and high‐performance thermocyclers.
Mobility Shift DNA‐Binding Assay Using Gel Electrophoresis
Authors: Buratowski Stephen , Chodosh Lewis A. 
Published: June 2004

AbstractDNA‐binding assay using nondenaturing polyacrylamide gel electrophoresis (PAGE) provides a simple, rapid, and extremely sensitive method for detecting sequence‐specific DNA‐binding proteins. Proteins that bind specifically to an end‐labeled DNA fragment retard the mobility of the fragment during electrophoresis, resulting in discrete bands corresponding to the individual protein‐DNA complexes. The assay described in this unit can be used to test binding of purified proteins or of uncharacterized factors found in crude extracts. This assay also permits quantitative determination of the affinity, abundance, association rate constants, dissociation rate constants, and binding specificity of DNA‐binding proteins. Three additional protocols describe a competition assay using unlabeled competitor DNA, an antibody supershift assay, and multicomponent gel shift assays.
Overview of Pharmacokinetics
Authors: Weber Steven J. 
Published: June 2004

AbstractPharmacokinetics is the study of the absorption, distribution, metabolism (biotransformation), and excretion (collectively referred to as ADME) of a compound following its administration. The physical/chemical state (e.g., molecular size and configuration, lipophilicity, and solubility) of a compound will greatly affect all these parameters. The aim of pharmacokinetic studies is to determine what dose and dosing schedule are sufficient to elicit and maintain a desired clinical response. This unit presents detailed procedures for dosing a rat via intravenous (i.v.), oral (p.o.), subcutaneous (s.c.), intraperitoneal (i.p.), and intramuscular (i.m.) routes. Protocols for blood collection, excreta collection, tissue collection, sample processing and calculation of data are also provided.
Determination of Compound Binding to Plasma Proteins
Authors: Dow Natasha 
Published: June 2004

AbstractThe pharmocokinetic and pharmacodynamic properties of a compound are profoundly affected by the extent of its binding to plasma proteins. Consequently, the determination of the plasma protein binding of a compound is essential during drug development and is increasingly required during lead prioritization. This unit describes the use of ultrafiltration and equilibrium dialysis techniques for determination of plasma protein binding. The pharmocokinetic and pharmacodynamic properties of a compound are profoundly affected by the extent of its binding to plasm
Autoradiographic Localization of Growth Factor Receptors in Neuronal Tissues
Authors: Doré Sylvain , Kar Satyabrata , Auld Daniel , Chabot Jean‐Guy , Quirion Remi 
Published: June 2004

AbstractThe physiological and pharmacological effects of growth factors, like those of neurotransmitters and neuromodulators, are mediated by binding to specific receptor sites. Using radiolabeled molecules with high specific activity, such as the endogenous ligand itself, it is possible to measure receptor binding in membrane preparations. This technique provides a means to characterize the selectivity and specificity of the receptors for a particular growth factor. Additionally, ligand‐binding assays make it possible to study in detail the structure‐activity relationships and mechanisms of action of growth factors. As an example of this methodology, this unit describes the use of both film and emulsion autoradiography to study the pharmacological characteristics and cellular distribution profile of insulin‐like growth factor receptors in the central nervous system. For the sake of clarity, in vitro film autoradiography in tissue sections and liquid emulsion autoradiography in cultured neurons are discussed separately. The physiological and pharmacological effects of growth factors, like those of neurotransmitters and neuromodulators, are medi.
Vigilance‐Controlled Quantified EEG in Safety Pharmacology
Authors: Dürmüller N. , Porsolt R.D. , Scherschlicht R. 
Published: June 2004

AbstractThe vigilance‐controlled quantified EEG can be used either as a safety, or as a discovery pharmacology procedure. Put strategically into the preclinical development process of a drug, it can be useful for making the decision about the future research direction to be taken. The experimental approach described in this unit is based on the rat EEG. Even though there are considerable differences in function and structure between human and rat brain, the EEG response to psychoactive drugs and convulsants is similar in the two species. Thus, the rat EEG is generally a reliable predictor for human CNS drug effects.
An Overview of QT Interval Assessment in Safety Pharmacology
Authors: Lacroix Pierre 
Published: June 2004

AbstractMedicinal products that prolong cardiac repolarization unintentionally, as assessed in terms of prolongation of the QT interval of the electrocardiogram, may trigger a potentially fatal arrhythmia called torsade de pointe (TDP). This lethal risk necessitates a robust preclinical evaluation before engaging in clinical trials. There are two different and complementary approaches to assess the potential of drugs to cause QT interval prolongation. The in vivo approach provides information on the potential of the compound to prolong the QT interval under near‐physiological conditions. It is mostly descriptive and not explanatory in terms of mechanisms of action. The in vitro approach provides much more mechanistic information, but is far removed from the clinical situation. Both approaches appear to possess reasonable predictive value, but the results obtained may largely depend on the experimental conditions used. This unit describes and presents an analytic strategy aimed at understanding the problems associated with this cardiovascular risk.
Overview of Electrophysiology
Authors: Bertrand Daniel 
Published: June 2004

AbstractThis unit provides an overview of the electrophysiological techniques used to determine the physiological and/or pharmacological properties of membrane ion channels at the single‐cell level. Only those techniques that measure potential changes at the transmembrane and membrane levels are discussed. The basics of current‐clamp recording, voltage‐clamp recording, and patch‐clamp procedures are presented along with their advantages and disadvantages. Additional topics are discussed with regard to their relation to electrophysiological measurements, such as membrane properties, ion selectivity, time resolution, ligand‐gated ion channels (LGICs), voltage‐dependent channels, and channel modulation and interactions. Also included is a discussion the electrophysiological properties of seven‐transmembrane‐domain receptors. This unit provides an overview of the electrophysiological techniques used to determine the physiological and/or pharmacologic of ion channels.
Electrophysiological Analysis of G Protein–Coupled Receptors in Mammalian Neurons
Authors: Proctor William R. , Dunwiddie Thomas V. 
Published: June 2004

AbstractThis unit describes general techniques that are useful for recording electrophysiological responses that are mediated via the activation of G‐protein coupled receptors (GPCRs). It includes a brief description of preparations, but focuses primarily on experiments using hippocampal brain slice preparations. Techniques for the preparation of brain slices, electrodes, filling solutions, and the recording protocols that are suitable for assessing the activity of GPCRs using electrophysiological techniques are summarized, and various protocols for the activation of these receptors are discussed.
Electrophysiological Analysis of Tetrodotoxin‐Resistant Sodium Channel Pharmacology
Authors: Dong Xiao‐Wei , Priestley Tony 
Published: June 2004

AbstractThis unit describes essential procedures to record the activity of and the effects of pharmacological manipulation upon tetrodotoxin‐resistant (TTXr) sodium channels. Readers are guided through the process of assembling a functional |patch‐clamp| electrophysiology apparatus and construction of software‐controlled voltage protocols that address various aspects of channel gating. Guidelines are also presented for analysis and display of the data typically generated by these experiments. For investigators wishing to study native channels in their appropriate cellular environment, protocols describing the isolation of viable cells from the main tissues that commonly express TTXr channels are also included.
Methods of Measuring Internalization of G Protein–Coupled Receptors
Authors: Evans Nicholas 
Published: June 2004

AbstractThis unit provides detailed protocols for measuring receptor internalization. The techniques are sufficiently generalized to be applicable to most receptors in a wide variety of cell types. Both radioactive and non‐radioactive techniques are described that may be used to quantify receptor internalization, and the differences between the two are highlighted. This unit discusses how quantification of internalization may be achieved, and the advantages and drawbacks of each technique. Low‐ and higher‐throughput methods are compared, and the technologies required to conduct the analyses are discussed.
Receptor Nomenclature Guidelines
Authors: Williams Michael 
Published: June 2004

AbstractReceptors are typically characterized via two distinct approaches: (1) the identification and pharmacological characterization of a receptor‐mediated response using classical pharmacological and/or radioligand approaches in tissues and animal models using selective agonist and antagonist ligands, and; (2) the cloning and expression of proteins with structural homology to known receptors, the function of which is subsequently established by studying the structure activity relationship (SAR) of receptor‐mediated responses. An additional means to characterize receptors proceeded, and evolved, with the structural approach, namely classification in terms of signal transduction mechanisms. The International Union of Pharmacology (IUPHAR) created guidelines and selected working groups for each receptor family to establish a common nomenclature system. Reports from those groups that have reached some degree of consensus have been summarized in this appendix.