RESUMO
The content of red pigment concentrating hormone (RPCH) in the eye-stalk of the crayfish Procambarus clarkii varies seasonally, with maximum values during the summer months and the lowest values in winter. The responsiveness of tegumentary chromatophores to synthetic RPCH varies concurrently.
Assuntos
Astacoidea/metabolismo , Hormônios de Invertebrado/metabolismo , Oligopeptídeos/metabolismo , Periodicidade , Animais , Olho/efeitos dos fármacos , Olho/metabolismo , Proteínas do Olho/biossíntese , Feminino , Hormônios de Invertebrado/farmacologia , Masculino , Oligopeptídeos/farmacologia , Ácido Pirrolidonocarboxílico/análogos & derivados , Pigmentos da Retina/biossíntese , Opsinas de Bastonetes , Estações do AnoRESUMO
In vibratome-cut slices from rat striatum and in the presence of 10 mM LiCl, the cholinergic agonist carbachol stimulated the accumulation of total [3H]inositol phosphates (EC50 11+/-1 microM and maximum effect 546+/-36% of basal). The response to 100 microM carbachol (497+/-24% of basal) was inhibited by muscarinic antagonists (1 microM), the rank order of efficacy being 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP; 100% inhibition) approximately pirenzepine (98+/-3%) > p-fluoro analog of hexahydro-sila-difenidol (pFHHSiD; 90+/-3%) >> methoctramine (32+/-7%) approximately tropicamide (30+/-10%). Antagonist inhibition curves best fit to a single-site model for 4-DAMP (pKi 8.9+/-0.2) whereas, for both pirenzepine and pFHHSiD, the best fit was to the two-site model. The pKi values for the high-affinity (8.3+/-0.2) and low-affinity (6.9+/-0.2) components for pirenzepine-mediated inhibition corresponded to those reported for M1 and M3 receptors, respectively. The pKi values for the high-affinity (8.2+/-0.3) and low-affinity (7.0+/-0.2) components for pFHHSiD inhibition were in good agreement with those reported for M3 and M1 receptors, respectively. Altogether these results indicate that carbachol-induced [3H]inositol phosphate formation in rat striatal slices is mediated by both M1 and M3 muscarinic receptors.
Assuntos
Carbacol/farmacologia , Corpo Estriado/efeitos dos fármacos , Fosfatos de Inositol/biossíntese , Antagonistas Muscarínicos/farmacologia , Animais , Corpo Estriado/metabolismo , Avaliação Pré-Clínica de Medicamentos , Técnicas In Vitro , Piperidinas/farmacologia , Pirenzepina/farmacologia , Ratos , Ratos WistarRESUMO
In this study, the authors set out to determine the presence of M3 muscarinic receptors in rat striatum by examining the binding of [3H]N-methylscopolamine ([3H]NMS) to striatal membranes and its displacement by antagonists with different affinity for M1 and M3 muscarinic receptors -pirenzepine; 4-diphenylacetoxy-N-methylpiperidine methiodide, 4-DAMP; and the p-fluoro analog of hexahydro-sila-difenidol, pFHHSiD). The specific binding of [3H]NMS to membranes from rat striatum (551 +/- 40 fmol.mg prot.-1, KD 0.11 +/- 0.01 nM) was displaced in a concentration-dependent manner by all three antagonists tested. Inhibition curves best fit to a single-site model for 4-DAMP (pKi 9.1 +/- 0.1), whereas for both pirenzepine and pFHHSiD, the best fit was to the two-site model. The pKi values for the high-affinity (8.0 +/- 0.2) and low-affinity (6.7 +/- 0.2) components for pirenzepine-mediated inhibition of [3H]NMS binding corresponded to those reported for M1 and M3 receptors, respectively. The pKi values for the high-affinity (7.7 +/- 0.1) and low-affinity (7.1 +/- 0.2) components for pFHHSiD inhibition were in good agreement with those reported for M3 and M1 receptors, respectively. Altogether, these results indicate the presence in rat striatum of both M1 and M3 muscarinic receptors. These findings might be relevant to the design and use of muscarinic antagonists in the treatment of neurological disorders such as Parkinson's disease.
Assuntos
Corpo Estriado/ultraestrutura , Receptores Muscarínicos/metabolismo , Animais , Ligação Competitiva , Cinética , Masculino , Antagonistas Muscarínicos/metabolismo , N-Metilescopolamina/metabolismo , Ratos , Ratos Wistar , Receptor Muscarínico M1 , Receptor Muscarínico M3 , TrítioRESUMO
In cross-chopped slices from rat thalamus and in the presence of 10 mM LiCl, noradrenaline stimulated the accumulation of [3H]inositol phosphates with [3H]inositol monophosphates ([3H]IP1) being the major product detected (86 +/- 2% of total [3H]inositol phosphates). Noradrenaline-induced [3H]IP1 accumulation was concentration-dependent and yielded and EC50 of 4.6 +/- 0.2 microM, maximum effect of 272 +/- 3% of basal formation and Hill coefficient (nH) of 1.6 +/- 0.1. The effect of 100 microM noradrenaline was inhibited by the alpha 1-adrenoceptor antagonists prazosin, (+)-niguldipine, 5-methylurapidil and WB-4101 (2-(2,6-dimethoxyphenoxyethyl) aminomethyl-1,4-benzodioxane). The inhibition curve for prazosin best fit to a single-site model whereas curves for (+)-niguldipine, 5-methylurapidil and WB-4101 best fit to a two-site model. The putative alpha 1D-adrenoceptor-selective antagonist BMY 7378 (8-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-8- azaspiro[4.5]decane-7,9-dione) showed low potency and efficacy to inhibit the response to noradrenaline. Pre-treatment of the slices with chloroethylclonidine (100 microM; 30 min) decreased by 64 +/- 4% the maximum response. Noradrenaline-induced [3H]IP1 accumulation was significantly reduced by Ca2+ removal (by 64 +/- 2%) and by the Ca(2+)-channel blockers Ni2+, Co2+ and nimodipine (inhibition of 56 +/- 6%, 54 +/- 5% and 41 +/- 5%, respectively). Taken together these results indicate that noradrenaline-induced inositol phosphate formation in thalamus slices is mainly mediated by the activation of both alpha 1B and alpha 1A subtypes of alpha 1-adrenoceptors.
Assuntos
Agonistas alfa-Adrenérgicos/farmacologia , Fosfatos de Inositol/biossíntese , Norepinefrina/farmacologia , Receptores Adrenérgicos alfa 1/metabolismo , Tálamo/metabolismo , Animais , Cálcio/farmacologia , Clonidina/análogos & derivados , Clonidina/farmacologia , Dioxanos/farmacologia , Técnicas In Vitro , Masculino , Ratos , Ratos Wistar , Receptores Adrenérgicos alfa 1/efeitos dos fármacos , Tálamo/efeitos dos fármacosRESUMO
In cross-chopped slices from rat thalamus and in the presence of 10 mM LiC1, the cholinergic agonist carbachol stimulated the accumulation of total [3H]inositol phosphates ([3H]IP2 = [3H]IP1 + [3H]IP2 + [3H]IP3). Best-fit values for the concentration-response curve for carbachol after 60 min incubation yielded an EC50 of 44 +/- 6 microM, maximum effect of 199 +/- 6% of basal accumulation and Hill coefficient (nH) of 1.1 +/- 0.1. Carbachol-induced [3H]IPs accumulation was inhibited by 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP; pKi 9.1) and the p-fluoro analogue of hexahydro-sila-difenidol (pF-HHSiD; pKi 8.1). Concentration-response curves for carbachol were shifted to the right in a parallel fashion by pirenzepine (100, 300 and 100 nM). A Schild plot of the data was linear (slope 0.95 +/- 0.04) and yielded a log KD for pirenzepine of -6.8 +/- 0.1. Taken together, these results suggest that carbachol-induced inositol phosphate accumulation in rat thalamus is mediated by muscarinic M3-receptors.
Assuntos
Carbacol/farmacologia , Fosfatos de Inositol/metabolismo , Agonistas Muscarínicos/farmacologia , Receptores Muscarínicos/metabolismo , Tálamo/metabolismo , Animais , Ligação Competitiva/fisiologia , Relação Dose-Resposta a Droga , Fosfatos de Inositol/biossíntese , Masculino , Antagonistas Muscarínicos/farmacologia , Técnicas de Cultura de Órgãos , Piperidinas/farmacologia , Pirenzepina/farmacologia , Ratos , Ratos Wistar , Tálamo/química , Tálamo/efeitos dos fármacos , TrítioRESUMO
To assess the usefulness of the South American primate Cebus apella as a model for neurovirulence of Junín virus, eight monkeys were inoculated with 10(5) LD50 of the attenuated XJ-Clone 3 Junín virus strain by the intrathalamic route. After the second week, weight loss and polyadenopathies were observed in most animals, one-half of which had a transient leukothrombocytopenia. Moderate clinical central nervous system (CNS) involvement was present in four of eight monkeys, while the rest had only mild neurologic signs. All recovered except one, which developed a deep coma and was killed in a pre-mortem stage at 18 days post-infection (pi). Junín virus was isolated from the throat from five, from the blood from three, and from the brain from two monkeys. In the most severely ill animal, virus titers higher than viremia were detected in both inoculated and contralateral brain hemispheres, as well as in lung, lymph node, and small intestine. Junín antigens and "in vivo" bound immunoglobulins were detected by immunofluorescence (IF) in the brain of four animals at 18, 21, 40, and 155 days pi. Moderate lymphocytic parenchymal and meningeal infiltration were observed in the brain of four animals, and gliosis was also present in the most affected monkey. Although the clinical response to infection was not uniform, all infected monkeys developed high IF antibodies. Cebus apella cannot be used as a highly sensitive model for Argentine hemorrhagic fever (AHF). However, the results obtained show that the XJ-Clone 3 strain can replicate in the primate CNS and to induce lesions and immunoglobulin deposition. In addition, viral persistence is suggested by the late detection of viral antigens in brain at 40 and 155 days pi.
Assuntos
Arenaviridae/patogenicidade , Arenavirus do Novo Mundo/patogenicidade , Encefalopatias/microbiologia , Cebidae , Cebus , Modelos Animais de Doenças , Febre Hemorrágica Americana/microbiologia , Animais , Anticorpos Antivirais/análise , Antígenos Virais/análise , Arenavirus do Novo Mundo/imunologia , Arenavirus do Novo Mundo/isolamento & purificação , Encéfalo/microbiologia , Feminino , Imunofluorescência , Viremia/microbiologia , VirulênciaRESUMO
The amino acid sequence of the L-CDR2 (complementarity-determining region) of Bla mRF (monoclonal rheumatoid factor) is identical to that of the Wa mRFs. The PSL2-CRI (crossreactive idiotype), as determined by anti-PSL2, which has been shown to be present on all Wa mRFs, is also present on the Bla mRF and other monoclonal autoantibodies. PSL2-CRI is, therefore, not unique to Wa mRFs and may be present on most IgM kappa monoclonal autoantibodies. Whether PSL2-CRI is a crossidiotype (XId) that is selectively present on autoantibodies or represents an allotypic marker for a V kappa III gene is undetermined.
Assuntos
Autoanticorpos/imunologia , Idiótipos de Imunoglobulinas/imunologia , Imunoglobulina M/imunologia , Fator Reumatoide/imunologia , Sequência de Aminoácidos , Anticorpos Monoclonais/imunologia , Reações Cruzadas , Humanos , Cadeias kappa de Imunoglobulina/imunologiaAssuntos
Animais Selvagens/imunologia , Anticorpos Antivirais/análise , Arenaviridae/imunologia , Arenavirus do Novo Mundo/imunologia , Febre Hemorrágica Americana/imunologia , Roedores/imunologia , Adolescente , Adulto , Animais , Animais Selvagens/microbiologia , Arenavirus do Novo Mundo/patogenicidade , Argentina , Criança , Pré-Escolar , Reservatórios de Doenças , Feminino , Cobaias , Febre Hemorrágica Americana/microbiologia , Humanos , Masculino , Camundongos , Pessoa de Meia-Idade , Roedores/microbiologiaAssuntos
Arenaviridae/patogenicidade , Arenavirus do Novo Mundo/patogenicidade , Cebidae , Cebus , Febre Hemorrágica Americana/microbiologia , Animais , Antígenos Virais/análise , Arenavirus do Novo Mundo/imunologia , Arenavirus do Novo Mundo/isolamento & purificação , Peso Corporal , Masculino , Contagem de PlaquetasRESUMO
The existence of antinuclear antibodies (ANA) and their relationship with anti-Trypanosoma cruzi specific agglutinins was studied in Rockland mice inoculated with 10(5), 10(4) or 10(3) epimastigotes and 2 X 10(5) trypomastigotes of T. cruzi. ANA and anti-T. cruzi agglutinins were detected at the same time in groups of mice receiving 10(5) culture forms, while in those groups of mice receiving 10(4) parasites agglutinins were established earlier than ANA which were present only at low titres. Positive serology and ANA were also detected in mice infected with trypomastigotes. Neither specific agglutinins nor ANA were found in groups of mice receiving 10(3) parasites probably because the number of parasites inoculated was too low to infect them. ANA were not found in not infected mice. ANA induced in mice by T. cruzi infection seem to be related to the amount of parasites inoculated and thus with the degree of the subsequent infection. Immunopathologic studies carried out in mice infected with trypomastigotes showed the presence of nodular deposits of gamma globulin in the kidneys, suggesting the possibility of an immune complex disease : the light and electron microscopic studies of the kidneys showed in the glomeruli an increase of the mesangium, hypercellularity and thickened basement membranes.
Assuntos
Anticorpos Antinucleares , Doença de Chagas/imunologia , Rim/imunologia , Trypanosoma cruzi/imunologia , Animais , Anticorpos Antinucleares/análise , Humanos , Doenças do Complexo Imune , Masculino , CamundongosRESUMO
An immunoadsorption technique was used to investigate the presence of antiglobulin factors in 28 patients with ankylosing spondylitis and 30 healthy individuals. Both groups were "sero-negative" by standard agglutination techniques. Whereas the controls were consistently negative, 18 of the 28 patients with ankylosing spondylitis had antiglobulins of the IgG class.
Assuntos
Anticorpos Anti-Idiotípicos/análise , Espondilite Anquilosante/imunologia , Adolescente , Adulto , Humanos , Imunoglobulina A/análise , Imunoglobulina G/análise , Imunoglobulina M/análise , Técnicas Imunológicas , Pessoa de Meia-Idade , gama-Globulinas/análiseRESUMO
Ultrastructural and immunohistochemical studies on tissues from five patients with Argentine hemorrhagic fever revealed previously undetected lesions caused by the viral infection. Two types of particle were seen in the cells of all organs examined. The particles had some characteristics similar to those described for arenaviruses. However, the virus-like particles were intracellular, had a single membrane, and apparently originated by a process of budding into the endoplasmic reticulum cisternae. Intranuclear bodies and three types of cytopolasmic change were observed in conjunction with the virus-like particles; Antigenic determinants of Junin virus were demonstrated in cells of all organs examined. Immunohistochemical experiments also indicated alterations in the cellular mechanisms of protein synthesis. Until now the pathogenesis of human diseases produced by arenaviruses has not been established. The results of this study suggest that in Argentine hemorrhagic fever the virus is responsible for a direct pathogenic action.