RESUMEN
Evidence on the action mechanisms of gold salts in the treatment of rheumatoid arthritis is still inconclusive. The intracellular localization of the place of action is likely. Therefore not only the serum gold levels but also the intracellular concentration of gold are of special interest. We measured the gold concentration in the serum and in the blood cells after in vitro application of aurothiomalate (Tauredon), gold keratinate (Auro-Detoxin) and triethylphosphine-gold (Ridaura) and in blood samples of patients undergoing these gold salts treatments. Cell-bound concentrations were found to vary extensively as a function of the gold compound used. While no or very little gold was present intracellularly after administration of the 2 parenteral drugs, up to 40% of the circulating gold was found to bind to the cells after administration of the triethylphosphine compound for gastro-intestinal absorption. The red cell concentration was more or less the same as that in the extracellular compartment. Gold apparently accumulated in the white cells, because the cell-bound concentration relative to unit volume was up to 20 times higher than the plasma level. The method used did not offer any information on the actual binding site of gold in white cells, i.e. cytoplasm versus nucleus versus cell membrane.
Asunto(s)
Oro/sangre , Compuestos Organometálicos , Fosfinas , Antiinflamatorios/sangre , Antiinflamatorios/uso terapéutico , Auranofina , Aurotioglucosa/análogos & derivados , Aurotioglucosa/sangre , Eritrocitos/análisis , Oro/uso terapéutico , Tiomalato Sódico de Oro/sangre , Humanos , Leucocitos/análisis , Compuestos Orgánicos de Oro , Compuestos Organofosforados/sangre , Péptidos/sangre , Espectrofotometría Atómica/métodosRESUMEN
We examined the binding of gold to serum proteins and the gold level in serum and in blood cells both after incubation in vitro and under longterm treatment with 3 gold compounds. Gold was measured by atomic absorption spectrophotometry, a serum fractionated by gel chromatography; blood cells were separated by gradient centrifugation, and intracellular gold was measured after pressure decomposition and MIBK-extraction. Aurothiomalate (GSTM), gold keratinate (GK), and auranofin (AF) were used. AF showed the highest binding to globulins in in vitro and in vivo. With GSTM and GK there were no measurable amounts of gold in the red cell lysate or erythrocyte membranes whereas with AF 40% of gold was localized within erythrocytes. The amount of gold in granulocytes and mononuclear cells after incubation with AF is considerably higher than with GK and GSTM.
Asunto(s)
Eritrocitos/análisis , Oro/sangre , Leucocitos/análisis , Auranofina , Aurotioglucosa/análogos & derivados , Aurotioglucosa/uso terapéutico , Oro/uso terapéutico , Tiomalato Sódico de Oro , Granulocitos/análisis , Técnicas In Vitro , Linfocitos/análisis , Monocitos/análisis , Péptidos/uso terapéutico , Distribución TisularAsunto(s)
Cnidarios/análisis , Péptidos , Toxinas Biológicas , Secuencia de Aminoácidos , Aminoácidos/análisis , Animales , Bioensayo , Braquiuros/efectos de los fármacos , Cromatografía en Gel , Cromatografía por Intercambio Iónico , Electroforesis en Gel de Poliacrilamida , Neuronas/efectos de los fármacos , Péptidos/aislamiento & purificación , Péptidos/farmacología , Toxinas Biológicas/aislamiento & purificación , Toxinas Biológicas/farmacologíaRESUMEN
Four toxic polypeptides, Toxins I, II, III and IV were isolated in pure form from the sea anemone Condylactis aurantiaca (Actinaria). Toxin isolation was achieved by alcoholic extraction of the homogenised sea anemones, batchwise adsorption onto cation exchangers, gel filtration on Sephadex G-50 and G-25 and ion-exchange chromatography on SP-Sephadex and QAE-Sephadex. Toxins from Condylactis aurantiaca all contain between 49 and 51 amino acids. Their amino acid compositions were compared to those of the Anemonia sulcata toxins. The toxins were tested on the shore crab Carcinus maenas by intramusclar injection. Crabs react highly sensitively to sea anemone toxins with muscle cramps and paralysis. For Condylactis toxins LD100 ranges from 2 to 6.6 mug/kg Carcinus maenas.
Asunto(s)
Cnidarios/análisis , Toxinas Marinas/análisis , Anémonas de Mar/análisis , Aminoácidos/análisis , Animales , Braquiuros , Toxinas Marinas/aislamiento & purificación , Péptidos/análisisRESUMEN
Sera from 28 untreated patients with Hodgkin's disease and from 120 healthy controls were investigated for the presence of circulating immune complexes using a modified 3% polyethylene glycol precipitation method with subsequent quantification of the precipitated protein. Elevated levels of precipitable protein were found in 79% (p less than 0.005) of Hodgkin's disease sera. The degree of elevation was associated with disease activity including the presence of B-symptoms. Constant and pronounced increase of precipitable protein was found in six patients with stage-III B nodular sclerosis subtype, thus exceeding the average amount of precipitable protein in healthy controls by a factor of 3-4. The erythrocyte sedimentation rate in 20 patients correlated with the amount of precipitable protein (r = 0.79). Additionally, partial component analysis of the precipitates was carried out by laser nephelometry. Immunoglobulins and complement components were identified as being major components of the precipitated material in sera both from patients and healthy controls, thus confirming the probability of the immune complex nature of the precipitates. Significant differences between patients and healthy controls concerned the amount of precipitable components. Elevation of precipitable IgM was found to be the most sensitive parameter (86% above means + 2 SD of normal controls, p less than 0.005). Increased amounts of precipitable IgG, C4, and Clq were found in 57-46% of patients' sera. Elevation of precipitable IgA and C3c were identified less often. The results suggest the quantification of precipitable immune complexes and their components to be of value as adjuncts in determining disease activity in Hodgkin's disease.
Asunto(s)
Complejo Antígeno-Anticuerpo/aislamiento & purificación , Enfermedad de Hodgkin/inmunología , Adolescente , Adulto , Anciano , Sedimentación Sanguínea , Precipitación Química , Niño , Complemento C4/análisis , Femenino , Humanos , Inmunoglobulina G/análisis , Inmunoglobulina M/análisis , Masculino , Persona de Mediana Edad , Estadificación de Neoplasias , Nefelometría y Turbidimetría , Polietilenglicoles , Probabilidad , Proteínas/análisisRESUMEN
The investigation of antibody kinetics following antigen-specific immunoadsorption in alkaline phosphatase immunized rats revealed significantly lower antibody levels than in untreated controls over a follow-up period of 6 weeks. A rebounding antibody synthesis as a result of specific depletion was not observed. Non-adsorption of specific antiidiotypic antibodies may explain these findings.
Asunto(s)
Fosfatasa Alcalina/inmunología , Anticuerpos/aislamiento & purificación , Antígenos/inmunología , Inmunización , Técnicas de Inmunoadsorción , Modelos Biológicos , Fosfatasa Alcalina/administración & dosificación , Animales , Anticuerpos/metabolismo , Especificidad de Anticuerpos , Bromuro de Cianógeno/química , Femenino , Ratas , Ratas Sprague-Dawley , Sefarosa/químicaRESUMEN
HISTORY AND CLINICAL FINDINGS: For nine years a 54-year-old woman had been suffering from worsening treatment-resistant cold-dependent purpura of the limbs as well as cutaneous ulcerations and arthralgia, which recently had occurred even at a even slight decrease in room temperature. INVESTIGATIONS: A special form of cryofibrinogenemia was identified by affinity-chromatographic separation of a plasma cryoprecipitate. From this cryoprecipitate a monoclonal antifibrinogen antibody (IgG-kappa) was isolated which, in the cold, formed a precipitating complex with fibrinogen. Paraproteinaemia was not demonstrated by conventional serum and plasma electrophoresis. There was no evidence of neoplasma. TREATMENT AND COURSE: Attempted treatment with steroids, fibrinolytic agents and intravenous cyclophosphamide was unsuccessful. But long-term repeated plasmaphereses and anti-immunoglobulin adsorption improved the symptoms. After 5 years of this treatment-14 years after onset of symptoms-the patient died of the consequences of fulminant pulmonary embolism. CONCLUSION: To establish the diagnosis of monoclonal cryofibrinogenemia it is necessary, first, to identify the cryoprecipitate in plasma; secondly, to undertake affinity-chromatographic separation of the cryoprecipitate with subsequent analysis of its components.
Asunto(s)
Anticuerpos Monoclonales/aislamiento & purificación , Autoanticuerpos/aislamiento & purificación , Fibrinógenos Anormales/inmunología , Púrpura/inmunología , Cromatografía de Afinidad , Crioglobulinas/inmunología , Resistencia a Medicamentos , Femenino , Fibrinógenos Anormales/metabolismo , Humanos , Persona de Mediana Edad , Plasmaféresis , Púrpura/tratamiento farmacológico , Úlcera Cutánea/inmunologíaRESUMEN
We report on a 54-year-old female patient with arthritis and a severe cold-induced leukocytoclastic vasculitis of the skin caused by a rare form of cryofibrinogenemia ("type II" cryofibrinogen). Affinity chromatography of cryoprecipitates from the patient's plasma revealed reversible cryoprecipitability of complexes composed of fibrinogen and a monoclonal antifibrinogen antibody (IgG3 kappa). Conventional serum and plasma electrophoresis did not detect the paraprotein. Control of symptoms was achieved by long-term plasmapheresis.