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Reference or citation managers aid in capturing and managing citations and associated full text, tracking references and citing them properly in manuscripts, and creating bibliographies. With more features than ever, selecting the most appropriate reference manager can be overwhelming for users and librarians. One common situation in which librarians are asked for advice involves building shared libraries of references to support collaborative group work. This project developed a structured evaluation for comparison of several common citation managers and prototypical use cases to help match features with user needs, preferences, and workflows. As products evolve and needs change, is there a "perfect fit"?
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Investigación Biomédica/organización & administración , Sistemas de Administración de Bases de Datos/organización & administración , Almacenamiento y Recuperación de la Información/métodos , Bibliotecólogos , Rol Profesional , Investigadores , Bases de Datos Bibliográficas , Humanos , Colaboración Intersectorial , North CarolinaRESUMEN
OBJECTIVE: Conventional anticonvulsants reduce neuronal excitability through effects on ion channels and synaptic function. Anticonvulsant mechanisms of the ketogenic diet remain incompletely understood. Because carbohydrates are restricted in patients on the ketogenic diet, we evaluated the effects of limiting carbohydrate availability by reducing glycolysis using the glycolytic inhibitor 2-deoxy-D-glucose (2DG) in experimental models of seizures and epilepsy. METHODS: Acute anticonvulsant actions of 2DG were assessed in vitro in rat hippocampal slices perfused with 7.5mM [K(+)](o), 4-aminopyridine, or bicuculline, and in vivo against seizures evoked by 6 Hz stimulation in mice, audiogenic stimulation in Fring's mice, and maximal electroshock and subcutaneous pentylenetetrazol (Metrazol) in rats. Chronic antiepileptic effects of 2DG were evaluated in rats kindled from olfactory bulb or perforant path. RESULTS: 2DG (10mM) reduced interictal epileptiform bursts induced by 7.5mM [K(+)](o), 4-aminopyridine, and bicuculline, and electrographic seizures induced by high [K(+)](o) in CA3 of hippocampus. 2DG reduced seizures evoked by 6 Hz stimulation in mice (effective dose [ED]50 = 79.7 mg/kg) and audiogenic stimulation in Fring's mice (ED50 = 206.4 mg/kg). 2DG exerted chronic antiepileptic action by increasing afterdischarge thresholds in perforant path (but not olfactory bulb) kindling and caused a twofold slowing in progression of kindled seizures at both stimulation sites. 2DG did not protect against maximal electroshock or Metrazol seizures. INTERPRETATION: The glycolytic inhibitor 2DG exerts acute anticonvulsant and chronic antiepileptic actions, and has a novel pattern of effectiveness in preclinical screening models. These results identify metabolic regulation as a potential therapeutic target for seizure suppression and modification of epileptogenesis.
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Anticonvulsivantes/uso terapéutico , Desoxiglucosa/uso terapéutico , Modelos Animales de Enfermedad , Epilepsia/tratamiento farmacológico , Hipocampo/efectos de los fármacos , Animales , Anticonvulsivantes/farmacología , Desoxiglucosa/farmacología , Relación Dosis-Respuesta a Droga , Electrochoque/métodos , Epilepsia/etiología , Epilepsia/patología , Epilepsia Refleja/tratamiento farmacológico , Epilepsia Refleja/etiología , Potenciales Evocados/efectos de los fármacos , Hipocampo/fisiopatología , Técnicas In Vitro , Masculino , Ratones , Pentilenotetrazol/toxicidad , Cloruro de Potasio/farmacología , Ratas , Ratas Sprague-Dawley , Factores de TiempoRESUMEN
The antiseizure activity of benzodiazepines (BDZs) 1-5 in mice and rats as animal models is described. These BDZs have selective efficacy for alpha2beta3gamma2 and alpha3beta3gamma2 GABA(A)-receptors. Significant anticonvulsant activity with little or no motor impairment and therapeutic indexes (TI) of 2.8-44 (mice, ip) were observed for compounds 2-4 in the subcutaneous metrazole seizure (scMET) test. In rats, orally (po) the TI was >5 to 105. These compounds represent novel leads in the search for anticonvulsants devoid of sedative, ataxic, and amnestic side effects.
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Anticonvulsivantes/síntesis química , Benzodiazepinas/síntesis química , Receptores de GABA-A/metabolismo , Animales , Anticonvulsivantes/farmacología , Anticonvulsivantes/toxicidad , Benzodiazepinas/farmacología , Benzodiazepinas/toxicidad , Convulsivantes , Hipocampo/efectos de los fármacos , Hipocampo/fisiopatología , Excitación Neurológica/efectos de los fármacos , Ligandos , Ratones , Pentilenotetrazol , Ratas , Convulsiones/inducido químicamente , Convulsiones/tratamiento farmacológico , Relación Estructura-ActividadRESUMEN
BACKGROUND/AIMS: GBR12909 has been reported to possess anticonvulsant activity with focal brain perfusion to the hippocampus of pilocarpine, although an earlier publication suggested any anticonvulsant effects were only mild. Here we further explored the anticonvulsant potential of GBR12909 with a suite of anticonvulsant assays in both zebrafish and mammals and then explored whether it possessed any QT effects which might limit clinical utility. METHODS: We assessed the anticonvulsant effects of GBR12909 in zebrafish pentylenetetrazole (PTZ), mammalian maximal electroshock and PTZ models of generalized epilepsy and a rodent hippocampal kindling model. Cardiac effects were assessed in zebrafish and man. RESULTS: GBR12909 possesses anticonvulsant activity in zebrafish and rodent models of generalized epilepsy. However, phase 1 human data indicated potential QT effects. Subsequent testing in a zebrafish QT assay confirmed marked arrhythmogenic potential. CONCLUSION: Further clinical development of GBR12909 in epilepsy was considered inappropriate because of insufficient window between the therapeutic effects and the cardiac arrhythmia problems identified in zebrafish assays. Any further development based on this mechanism of action should avoid the GBR12909 chemical scaffold, or involve structure-activity dissociation of its neurological and cardiac effects.
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Anticonvulsivantes/farmacología , Inhibidores de Captación de Dopamina/farmacología , Epilepsia/tratamiento farmacológico , Canales de Potasio Éter-A-Go-Go/metabolismo , Miocardio/metabolismo , Piperazinas/farmacología , Animales , Anticonvulsivantes/efectos adversos , Arritmias Cardíacas/inducido químicamente , Perros , Inhibidores de Captación de Dopamina/efectos adversos , Electrocardiografía , Masculino , Ratones , Piperazinas/efectos adversos , Ratas , Ratas Sprague-Dawley , Pez CebraRESUMEN
Geriatric epilepsy is a significant clinical problem that has not been studied adequately in animal models. This chapter will review the available literature with particular attention to models that have demonstrated how acute seizures and epilepsy in aged animals differ from those of younger animals. Studies include several strains of mice [e.g., El, DBA, senescence-accelerated mouse (SAM), Cacnb4 knockout] as well as acute seizure models in common strains of aged mice. Aged rats (including Fischer 344, Wistar, and Sprague-Dawley) have been used in acute seizure, lesion, and epilepsy models. This area of research remains largely unexplored and therefore provides numerous opportunities for new investigations.
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Modelos Animales de Enfermedad , Epilepsia , Anciano , Anciano de 80 o más Años , Animales , Gerbillinae , Humanos , Ratones , RatasRESUMEN
Ggamma11 is an unusual guanine nucleotide-binding regulatory protein (G protein) subunit. To study the effect of different Gbeta-binding partners on gamma11 function, four recombinant betagamma dimers, beta1gamma2, beta4gamma2, beta1gamma11, and beta4gamma11, were characterized in a receptor reconstitution assay with the G(q)-linked M1 muscarinic and the G(i1)-linked A1 adenosine receptors. The beta4gamma11 dimer was up to 30-fold less efficient than beta4gamma2 at promoting agonist-dependent binding of [35S]GTPgammaS to either alpha(q) or alpha(i1). Using a competition assay to measure relative affinities of purified betagamma dimers for alpha, the beta4gamma11 dimer had a 15-fold lower affinity for G(i1) alpha than beta4gamma2. Chromatographic characterization of the beta4gamma11 dimer revealed that the betagamma is stable in a heterotrimeric complex with G(i1) alpha; however, upon activation of alpha with MgCl2 and GTPgammaS under nondenaturing conditions, the beta4 and gamma11 subunits dissociate. Activation of purified G(i1) alpha:beta4gamma11 with Mg+2/GTPgammaS following reconstitution into lipid vesicles and incubation with phospholipase C (PLC)-beta resulted in stimulation of PLC-beta activity; however, when this activation preceded reconstitution into vesicles, PLC-beta activity was markedly diminished. In a membrane coupling assay designed to measure the ability of G protein to promote a high-affinity agonist-binding conformation of the A1 adenosine receptor, beta4gamma11 was as effective as beta4gamma2 when coexpressed with G(i1) alpha and receptor. However, G(i1) alpha:beta4gamma11-induced high-affinity binding was up to 20-fold more sensitive to GTPgammaS than G(i1) alpha:beta4gamma2-induced high-affinity binding. These results suggest that the stability of the beta4gamma11 dimer can modulate G protein activity at the receptor and effector.
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Subunidades beta de la Proteína de Unión al GTP/metabolismo , Subunidades gamma de la Proteína de Unión al GTP/metabolismo , Isoenzimas/metabolismo , Subunidades de Proteína/metabolismo , Receptor de Adenosina A1/metabolismo , Receptor Muscarínico M1/metabolismo , Fosfolipasas de Tipo C/metabolismo , Animales , Anticuerpos/inmunología , Sitios de Unión , Precipitación Química , Dimerización , Subunidades alfa de la Proteína de Unión al GTP Gq-G11/aislamiento & purificación , Subunidades beta de la Proteína de Unión al GTP/aislamiento & purificación , Subunidades gamma de la Proteína de Unión al GTP/aislamiento & purificación , Fosfolipasa C beta , Unión Proteica , Isoformas de Proteínas/metabolismo , Proteínas Recombinantes/metabolismo , Spodoptera , Termodinámica , PavosRESUMEN
Allosteric enhancers (AEs) of the A(1) adenosine receptor (A(1)AR) have potential as drugs for treating neurological, cardiovascular, and renal diseases. This report describes the synthesis and evaluation of a series of 6-aryl-8H-indeno[1,2-d]thiazol-2-ylamines that exhibited AE activity at the A(1)AR. Palladium-mediated condensation of arylboronic acids with 5-bromoindan-1-one generated arylindanones 2a-aj for iodine-catalyzed condensation with thiourea, generating 2-aminothiazolium salts 3a-aj. Binding studies using membranes from cells stably expressing human A(1)ARs, A(2A)ARs, or A(3)ARs evaluated AE activity and receptor subtype selectivity. The EC(50) of the AE activities of compounds 3m-o, 3x, and 3ae were 2.2, 1.5, 0.9, 1.0, and 3.0 muM, respectively, substantially lower than that of the well characterized 2-amino-3-aroylthiophene (PD 81,723), >10 microM. The new compounds also have substantially higher maximal AE activity. These compounds had no AE activity at the A(2A)AR and only minimal activity at the A(3)AR.
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Agonistas del Receptor de Adenosina A1 , Aminas/síntesis química , Indenos/síntesis química , Tiazoles/síntesis química , Antagonistas del Receptor de Adenosina A1 , Regulación Alostérica , Aminas/química , Aminas/farmacología , Animales , Unión Competitiva , Línea Celular , Cricetinae , Cricetulus , Humanos , Indenos/química , Indenos/farmacología , Ensayo de Unión Radioligante , Relación Estructura-Actividad , Tiazoles/química , Tiazoles/farmacologíaRESUMEN
The A(2A) adenosine receptor (A(2A)AR) mediates anti-inflammatory actions of adenosine in a variety of cell types. LPS (lipopolysaccharide) was reported to induce a small (<2-fold) increase in the expression of A(2A)AR mRNA in human monocytes and monocytic cell lines. We investigated the effects of LPS on the expression of adenosine receptor mRNAs in primary mouse IPMPhi (intraperitoneal macrophages), human macrophages and Wehi-3 cells. Treatment with 10 ng/ml LPS for 4 h produced a >100-fold increase in A(2A)AR mRNA. LPS-induced increases in mRNA for A(2A)AR and TNFalpha (tumour necrosis factor alpha) are reduced by 90% in IPMPhi pretreated with the NF-kappaB (nuclear factor kappaB) inhibitor, BAY 11-7082 {(E)3-[(4-methylphenyl)sulphonyl]-2-propenenitrile; 10 microM}. In Wehi-3 cells exposed to LPS, A(2A)AR and A(2B)AR transcripts are elevated by 290- and 10-fold respectively, the A(1)AR transcript is unchanged and the A(3)AR transcript is decreased by 67%. The induction of A(2A)AR mRNA by LPS is detectable after 1 h, reaches a peak at 6 h at 600 times control and remains elevated beyond 24 h. The ED50 (effective dose) of LPS is 2.3 ng/ml. A(2A)AR receptor number, measured by 125I-ZM241385 binding to whole cells, is undetectable in naïve cells and increases linearly at a rate of 23 receptors x cell(-1) x min(-1) to a B(max) of 348 fmol/mg (28000 receptors/cell) in 20 h. The increase in receptor number is correlated with an increase in the potency of an A(2A) agonist (4-{3-[6-amino-9-(5-ethylcarbamoyl-3,4-dihydroxy-tetrahydro-furan-2-yl)-9H-purin-2-yl]-prop-2-ynyl}-cyclohexanecarboxylic acid methyl ester; referred to as ATL146e) to stimulate cAMP in these cells. After LPS pretreatment, the potency of the A(2A) agonist, ATL146e, to reduce TNFalpha release from IPMPhi was increased by 200-fold. The results support the hypothesis that regulation of adenosine receptor expression, especially up-regulation of the A(2A)AR, is part of a delayed feedback mechanism initiated through NF-kappaB to terminate the activation of human and mouse macrophages.
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Regulación de la Expresión Génica/efectos de los fármacos , Lipopolisacáridos/farmacología , Macrófagos/efectos de los fármacos , FN-kappa B/metabolismo , Receptor de Adenosina A2A/genética , Transcripción Genética/efectos de los fármacos , Animales , Células Cultivadas , Ácidos Ciclohexanocarboxílicos/farmacología , Humanos , Macrófagos/metabolismo , Ratones , Nitrilos/farmacología , Unión Proteica , Purinas/farmacología , ARN Mensajero/genética , ARN Mensajero/metabolismo , Sulfonas/farmacología , Triazinas/farmacología , Triazoles/farmacología , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
This report describes the synthesis and structure-activity relationships of a new class of A(1) adenosine receptor agonist allosteric enhancers, 2-aminothiazolium salts. The EC(50) of compounds 6a, 6b, 7, and 8 were 0.3, 4.5, 3.8, and 1.2 microM, substantially lower than that of the 'Gold Standard' 2-amino-3-benzoyl thiophene (PD 81,723), which has an EC(50) of 38 microM.
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Agonistas del Receptor Purinérgico P1 , Tiazoles/farmacología , Regulación Alostérica , Humanos , Relación Estructura-Actividad , Tiazoles/síntesis química , Tiazoles/químicaRESUMEN
Agonists bind with higher affinity to G protein-coupled heptahelical receptors than to uncoupled receptors. Recombinant A(1) and A(3) adenosine receptors couple well to G(i/o), but recombinant human A(2A) adenosine receptors (hA(2A)AR) couple poorly to G(s) and bind agonists with K(i) values in binding assays that are much higher than ED(50) values for functional responses such as coronary dilation and inhibition of neutrophil oxidative burst. In this study, we produced hA(2A)AR-G protein complexes in membranes derived from Sf9 cells quadruply infected with receptors and heterotrimeric G protein subunits. The composition of G(beta) markedly influences coupling such that A(2A)AR-alpha(s)beta(1)gamma(2) are 8 +/- 2% coupled whereas equivalently expressed A(2A)AR-alpha(s)beta(4)gamma(2) are 40 +/- 2% coupled. Hence, we were able for the first time to accurately measure high-affinity agonist binding to hA(2A)AR. The agonist 2-[2-(4-amino-3-[(125)I]iodophenyl)ethylamino]adenosine binds to coupled and uncoupled hA(2A)AR with K(D) values of 0.46 nM and 26 nM, respectively, a difference in affinity of 57-fold. The addition of GTPgammaS converts all receptors to the low-affinity state. A(2A)AR coupling does not influence binding of antagonists including, (125)I-4-(2-[7-amino-2-[2-furyl][1,2,4]triazolo[2,3-a][1,3,5]triazin-5-yl-amino]ethyl)phenol ((125)I-ZM241385), K(D) = 0.5 nM. Based on a comparison of high-affinity binding sites, N(6)-3-iodo-2-chlorobenzyladenosine-5'-N-methyluronamide is only 8-fold A(3) selective (A(2A Ki, H) = 18.3 +/- 3.2 nM; A(3 Ki, H) = 2.4 +/- 0.3 nM) and 2-chloro-N(6)-cyclopentyladenosine is only 33-fold A(1) selective (A(2A Ki, H) = 11.0 +/- 1.9; A(1 Ki, H) = 0.3 +/- 0.1). We conclude that recombinant hA(2A)AR can form a high-affinity receptor-G protein complex with alpha(s)beta(4)gamma(2) that is useful for determining receptor selectivity.
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Proteínas de Unión al GTP Heterotriméricas/metabolismo , Receptores Purinérgicos P1/metabolismo , Animales , Western Blotting , Células Cultivadas , Humanos , Insectos , Conformación Proteica , Ensayo de Unión Radioligante , Receptor de Adenosina A2A , Receptores Purinérgicos P1/química , Receptores Purinérgicos P1/genética , Proteínas Recombinantes/metabolismoRESUMEN
2-Amino-3-benzoylthiophenes are allosteric enhancers (AE) of agonist activity at the A(1) adenosine receptor. The present report describes syntheses and assays of the AE activity at the human A(1)AR (hA(1)AR) of a panel of compounds consisting of nine 2-amino-3-aroylthiophenes (3a-i), eight 2-amino-3-benzoyl-4,5-dimethylthiophenes (12a-h), three 3-aroyl-2-carboxy-4,5-dimethylthiophenes (15a-c), 10 2-amino-3-benzoyl-5,6-dihydro-4H-cyclopenta[b]thiophenes (17a-j), 14 2-amino-3-benzoyl-4,5,6,7-tetrahydrobenzo[b]thiophenes (18a-n), and 15 2-amino-3-benzoyl-5,6,7,8-tetrahydro-4H-cyclohepta[b]thiophenes (19a-o). An in vitro assay employing the A(1)AR agonist [(125)I]ABA and membranes from CHO-K1 cells stably expressing the hA(1)AR measured, as an index of AE activity, the ability of a candidate AE to stabilize the agonist-A(1)AR-G protein ternary complex. Compounds 3a-i had little or no AE activity, and compounds 12a-h had only modest activity, evidence that AE activity depended absolutely on the presence of at least a methyl group at C-4 and C-5. Compounds 17a-c lacked AE activity, suggesting the 2-amino group is essential. Polymethylene bridges linked thiophene C-4 and C-5 of compounds 17a-j, 18a-n, and 19a-o. AE activity increased with the size of the -(CH(2))(n)()- bridge, n = 3 < n = 4 < n = 5. The 3-carbethoxy substituents of 17a, 18a, and 19a did not support AE activity, but a 3-aroyl group did. Bulky (or hydrophobic) substituents at the meta and para positions of the 3-benzoyl group and also 3-naphthoyl groups greatly enhanced activity. Thus, the hA(1)AR contains an allosteric binding site able to accommodate 3-aroyl substituents that are bulky and/or hydrophobic but not necessarily planar. A second region in the allosteric binding site interacts constructively with alkyl substituents at thiophene C-4 and/or C-5.