RESUMEN
BACKGROUND: The major allergen of Baltic cod (Gadus callarias) is a 12.3-kDa parvalbumin with two calcium-binding sites corresponding to EF-hand motifs. Our group found a 24-kDa IgE-reactive band that was also recognized by a monoclonal antiparvalbumin antibody in Atlantic cod (Gadus morhua). Our purpose was to purify and to determine the cDNA deduced sequence of this new cod allergen. METHODS: Proteins from pre rigor mortis Atlantic cod were separated by gel filtration and the eluted peaks were analysed by SDS-PAGE and Western blotting with sera of sensitized patients and with antiparvalbumin. Protein bands were microsequenced, RNA transcripts were amplified by reverse transcription and polymerase chain reaction (RT-PCR) using primer combinations overlapping the open reading frame. RESULTS: Four IgE and antiparvalbumin reactive proteins(12.5, 24, 38 and 51 kDa) were detected in gel filtration eluate. The cDNA deduced sequence of the 24 kDa protein had 109 amino acid residues with a molecular weight of 11.5 kDa and a theoretical pI of 4.34. The 24 kDa band corresponded therefore to a dimer of a beta-parvalbumin. Its homology was higher with Sal sI than with Gad cI. This new allergen was named Gad mI. CONCLUSION: We have characterized a new parvalbumin allergen in Gadus morhua. This protein formed oligomers in native and in reducing conditions. Gad mI and Gad cI may correspond to two distinct genes of Gadus species.
Asunto(s)
Alérgenos/genética , ADN Complementario/genética , Proteínas de Peces/genética , Peces/inmunología , Parvalbúminas/genética , Alérgenos/inmunología , Animales , Western Blotting , Proteínas de Peces/inmunología , Hipersensibilidad a los Alimentos/etiología , Hipersensibilidad a los Alimentos/inmunología , Humanos , Parvalbúminas/inmunología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ADN , Análisis de Secuencia de ProteínaRESUMEN
OBJECTIVE: to determine whether inhaled nitric oxide (NO) would alter leukocyte kinetics in the septic microvasculature. DESIGN: Randomized, controlled trial. SETTING: Experimental laboratory. SUBJECTS: Male Sprague Dawley rats. INTERVENTIONS: Rats were treated with either saline or endotoxin (10 mg/kg, iv) and then allowed to breathe either air or air plus NO (10 ppm). MEASUREMENTS AND MAIN RESULTS: After a 4-hr period, rolling, firm adhesion, and emigration of leukocytes and endothelial dysfunction were monitored in mesenteric venules by using intravital videomicroscopy. Compared with controls, endotoxemic rats exhibited a profound influx in mesenteric venule rolling leukocytes (55+/-17 vs. 70+/-19 leukocytes/min; p < .05), associated with a reduction of leukocyte rolling velocity (83+/-14 vs. 34+/-3 microm/sec; p < .05). In endotoxemic rats, venular endothelium leukocyte firm adhesion (1.15+/-0.32 vs. 4.08+/-0.96 leukocytes/ 100 microm; p < .05) and emigration (0.84+/-0.47 vs. 4.23+/-1.2 leukocytes/100 microm; p < .05) increased compared with controls. Inhaled NO had no effect on leukocyte kinetics in control rats. Inhaled NO significantly attenuated endotoxin-induced venular endothelium leukocyte adhesion (4.08+/-0.96 vs. 1.86+/-0.76 leukocytes/100 microm; p < .05) and emigration (4.23+/-1.2 vs. 1.68+/-0.72 leukocytes/100 microm; p < .05). Compared with control rats, macromolecular (FITC-dextran) vascular leakage, expressed as the perivenular/intravenular fluorescence intensity ratio, increased in endotoxemic rats (0.56+/-0.02 vs. 0.81+/-0.05; p < .01). Endotoxin-induced macromolecular vascular leakage increases were partially prevented by inhaled NO (0.66+/-0.01 vs. 0.56+/-0.02; p < .05). CONCLUSION: These observations suggest that inhaled NO reduces leukocyte adhesion and the degree of vascular permeability dysfunction in mesenteric venule of endotoxemic rats.
Asunto(s)
Endotoxemia/tratamiento farmacológico , Infecciones por Escherichia coli/tratamiento farmacológico , Leucocitos/efectos de los fármacos , Venas Mesentéricas/efectos de los fármacos , Óxido Nítrico/administración & dosificación , Vasodilatadores/administración & dosificación , Administración por Inhalación , Animales , Permeabilidad Capilar/efectos de los fármacos , Adhesión Celular/efectos de los fármacos , Evaluación Preclínica de Medicamentos , Endotoxemia/sangre , Endotoxemia/fisiopatología , Infecciones por Escherichia coli/sangre , Infecciones por Escherichia coli/fisiopatología , Hemodinámica/efectos de los fármacos , Cinética , Leucocitos/fisiología , Masculino , Venas Mesentéricas/fisiopatología , Óxido Nítrico/farmacología , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Vasodilatadores/farmacología , Vénulas/efectos de los fármacos , Vénulas/fisiopatologíaRESUMEN
OBJECTIVES: Nitric oxide is known to prevent platelet aggregation and clot formation. Inhibitors of nitric oxide synthase might promote or enhance endotoxin disseminated intravascular coagulation. The present study was designed to evaluate the effects of the arginine analog, N omega-nitro-L-arginine methyl ester (L-NAME), on the endotoxin-induced disseminated intravascular coagulation in a porcine model of septic shock. DESIGN: Prospective, comparative, experimental study. SETTING: Laboratory at a large university hospital. SUBJECTS: Sixteen female piglets, weighing 20 to 28 kg. INTERVENTIONS: Three groups of animals were studied: a control group (n = 6); a lipopolysaccharide (LPS)-treated group (n = 5) receiving Escherichia coli endotoxin (5 micrograms/kg/min over 30 mins); and an LPS + L-NAME group (n = 5) receiving endotoxin and, 1 hr after, a bolus of L-NAME (25 mg/kg). MEASUREMENTS AND MAIN RESULTS: Hemodynamic changes, usual coagulation parameters, and plasma concentrations of thrombin-antithrombin complexes, antithrombin III activity (At III), tissue plasminogen activator, plasminogen activator inhibitor type 1, and von Willebrand factor were measured at baseline, and at 30, 60, 90, 120, 180, 240, and 300 mins. After euthanasia or death, lungs and kidneys were withdrawn for histologic study. The extent of microvascular thrombosis was assessed by a semiquantitative disseminated intravascular coagulation score. In both septic endotoxin group, administration of LPS resulted in hemodynamic changes typical of severe septic shock, with disseminated intravascular coagulation and histologic changes characterized by adult respiratory distress syndrome and kidney microthrombosis. L-NAME administration normalized mean arterial pressure with a dramatic increase in systemic vascular resistances and a marked decrease in cardiac index. The changes in usual coagulation parameters, AT III, tissue plasminogen activator, and plasminogen-activator inhibitor type 1 concentrations were not different between both septic groups. However, in the LPS + L-NAME group, thrombin-antithrombin complexes and von Willebrand factor were higher and associated with a higher histologic disseminated intravascular coagulation score. CONCLUSION: In this model of endotoxin septic shock, L-NAME administration resulted in histologic and coagulation changes consistent with an increased activation of intravascular coagulation.
Asunto(s)
Coagulación Intravascular Diseminada/tratamiento farmacológico , Coagulación Intravascular Diseminada/microbiología , Inhibidores Enzimáticos/uso terapéutico , NG-Nitroarginina Metil Éster/uso terapéutico , Choque Séptico/complicaciones , Animales , Modelos Animales de Enfermedad , Coagulación Intravascular Diseminada/patología , Evaluación Preclínica de Medicamentos , Escherichia coli , Femenino , Hemodinámica/efectos de los fármacos , Lipopolisacáridos , Índice de Severidad de la Enfermedad , PorcinosAsunto(s)
Petróleo/envenenamiento , Adulto , Preescolar , Femenino , Humanos , Lactante , Masculino , Edema Pulmonar/inducido químicamente , Edema Pulmonar/terapiaRESUMEN
During acute intoxications rhabdomyolysis appear with a great clinical polymorphism. The muscular involvement is not always evident because of its shortness and latence. Practically the problem is one of localised muscle damage, hyperkaliema or acute renal insufficiency. The serum isoenzymes of CPK levels, the presence of myoglobinuria are necessary for the diagnosis. The most serious rhabdomyolysis depend on the added injury of respiratory muscles and myocardium. Many toxic substances can involve rhabdomyolysis but the most frequent ones are sedatives, carbonic oxyde, ethanol. Only the complications are treated.
Asunto(s)
Mioglobinuria/inducido químicamente , Intoxicación , Intoxicación Alcohólica/complicaciones , Anestesia General/efectos adversos , Animales , Intoxicación por Monóxido de Carbono/complicaciones , Enfermedades Transmitidas por los Alimentos/complicaciones , Glycyrrhiza , Heroína/envenenamiento , Humanos , Hipnóticos y Sedantes/envenenamiento , Músculos/patología , Mioglobinuria/diagnóstico , Mioglobinuria/patología , Plantas Medicinales , Codorniz , Succinilcolina/efectos adversosRESUMEN
Among 17 septicemic patients, a statistically decrease of seric oligo-elements is demonstrated. The longitudinal study shows the rapid correction of these perturbations in the surviving group and the lack of correction among the non survivors. The level of phosphorus is the most interesting one. The importance of S.R.E. activity is probably the physiopathologic support of seric oligo-element modifications in sepsis.
Asunto(s)
Sepsis/sangre , Oligoelementos/sangre , Adolescente , Adulto , Anciano , Calcio/sangre , Cobre/sangre , Femenino , Humanos , Hierro/sangre , Masculino , Persona de Mediana Edad , Fósforo/sangre , PronósticoAsunto(s)
Oxigenoterapia Hiperbárica , Hipoxia Encefálica/terapia , Intento de Suicidio , Accidentes , Adolescente , Adulto , Anciano , Niño , Femenino , Humanos , Hipoxia Encefálica/etiología , Masculino , Persona de Mediana EdadRESUMEN
On the basis of 47 cases of gas gangrene collected over the three year period between 1974 and 1976, the authors review the circumstances surrounding its development, the clinical features and the prognosis of the disorder which remains grave despite a well-defined therapeutic protocol combining surgery, antibiotics and hyperbaric oxygen. There would appear to be a real resurgence of the disease at the present time. Post-traumatic and surgical aetiologies predominate, giving rise to two types of gangrene: clostridial gas gangrene secondary to contamined wounds, with a quasi-constant vascular element, affecting predominantly the limbs, and nonclostridrial gangrene, the increasing prevalence of which involves essentially spetic abdomino-pelvic surgery. In the light of this study, prognosis would appear to be related to the underlying terrain in which the gangrene occurs, to certain features of the clinical picture and, above all, to the possibilities of early application of the complete therapeutic protocol. Strict prophylactic measures would alone seem capable of preventign the worrying increase in the number of cases of gas gangrene.