The aim was to analyze the effect of leukotriene synthesis inhibitor administered intraperitoneally in vasopressin release during sepsis. Male Wistar rats received injections of MK-886 (1.0, 2.0 or 4.0 mg/kg) or vehicle (DMSO 5%) 1 h before cecal ligation and puncture. There was some variation on the survival rate depending on the dose used but the drug did not modify the hematocrit, osmolality, serum sodium and nitrate, plasma protein, and neutrophil recruitment, in any dose. Nevertheless, vasopressin (AVP) release decreased in a dose-response manner in the early phase of sepsis. These results support the suggestion that leukotrienes (LTs) are involved in AVP release during sepsis.
Shrimp was obtained from the sea by fisherman, who were instructed to not add any additives and to place the shrimp at -20oC immediately after catching. Shrimp was kept at -20oC, and at the time of use, shrimp was thawed, had their shell removed, and boiled in water for 10 minutes.For the double-blind-placebo-controlled shrimp challenge, shrimp was crushed using a food processor, and added to a mixture of hazelnut cream, oat flakes, milk-based ice cream with small bits of chocolate, powdered chocolate and vanilla extract, containing 15.4 % of fat. Prior to challenge, patients were asked about any reactions to each of the components of the mixture. The cumulative dose of 1g shrimp protein was the maximum we could use to allow shrimp taste to be disguised in the food matrix. Placebo challenges were carried out using the same mixture minus shrimp and similar timing schedule. For the open challenge, cooked shrimp prepared as described for the DBPCFC was lightly salted and seasoned with olive oil. Active and placebo challenges were performed one week apart.
IgE ImmunoblotsNatural Lit v 1 and recombinant Per a 7 were prepared in our laboratory according to previously published methods (Santos et al). Protein concentration of shrimp extract, Lit v 1 and Per a 7 was measured by Bradford assay, using Bovine Serum Albumin (BSA) as standard. Fifty micrograms of protein/lane were applied to 10% Sodium Dodecyl Sulfate Polyacrylamide Electrophoresis (SDS-PAGE) gels. Electrophoresis was carried out under reducing conditions with betamercaptoethanol. Protein transferring to nitrocellulose membranes (Millipore, Bedford, MA) was confirmed by Ponceau S staining. Membranes were blocked with Phosphate Buffered Saline (PBS) + 0.1% Tween + 5% skim milk (blocking solution) for 1 hour at room temperature. After
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