Differences between the real and the desired worlds in the results of clinical trials

OBJECTIVE:We refer to the effectiveness (known as pragmatic or real world) and efficacy (known as explanatory or desired or ideal world) of interventions. However, these terms seem to be randomly chosen by investigators who design clinical trials and do not always reflect the true purpose of the study. A pragmatic-explanatory continuum indicator summary tool was thus developed with the aim of identifying the characteristics of clinical trials that distinguish between effectiveness and efficacy issues. We verified whether clinical trials used the criteria proposed by the indicator summary tool, and we categorized these clinical trials according to a new classification.METHOD:A systematic survey of randomized clinical trials was performed. We added a score ranging from 0 (more efficacious) to 10 (more effective) to each domain of the indicator summary tool and proposed the following classifications: high efficacy

Effect of polymerization with transglutaminase on in vitro digestion and antigenicity of beta-lactoglobulin

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)The in vitro digestibility of beta-Lactoglobulin (beta-Lg) denatured by heat or chemical (0.1 or 0.25 mol L-1 Cys) treatments, polymerized or not by transglutaminase (10 or 25 U g(-1) protein), and the antigenicity of the products before and after in vitro digestion, was investigated. The polymerization was more efficient in the presence of Cys than after heat treatment. The addition of Cys, associated or not with polymerization, increased digestibility by more than 20% as compared to untreated beta-Lg. The pepsin digest of the polymerized beta-Lg showed lower reactivity towards IgE than the untreated or denatured beta-Lg. After digestion by pepsin and pancreatin, untreated and heat treated beta-Lg still showed antigenic activity, whereas the products released from beta-Lg denatured by Cys and from the polymerized samples showed no reactivity towards IgE. The polymerization of chemically denatured beta-Lg facilitated the action of the gastrointestinal enzymes, and their digestion products presented low antigenic properties. (C) 2012 Elsevier Ltd. All rights reserved.252123131Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)CNPq [479322/2008-2

The effect of transglutaminase-induced polymerization in the presence of cysteine on beta-lactoglobulin antigenicity

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)The effect of polymerization by the enzyme transglutaminase (TG) on the antigenicity of beta-lactoglobulin (beta-Lg) was investigated. Polymerization was carried out using 7% heat treated beta-Lg and 5-50 U TG g(-1), substrate or 7% untreated beta-Lg in the presence of 0.05-0.4 mol L(-1) cysteine (Cys) and 25 U TG g(-1) substrate. The electrophoretic profile of polymerized samples showed bands corresponding to high molecular mass. For antigenicity evaluation, sera from BALB/c mice sensitized with native beta-Lg, beta-Lg polymerized by 25 U TG g(-1) (beta-Lg HT TG) or polymerized in 0.25 mol L(-1) Cys (beta-Lg Cys TG) were used. Animals sensitized with beta-Lg Cys TG showed lower levels of IgG1 and IgE than those immunized with native beta-Lg or beta-Lg HT TG. These results suggested that polymerization in the presence of Cys modified or hid epitopes, reducing the potential antigenicity of beta-Lg, whereas heat treatment followed by polymerization did not lead to a reduction in antigenicity. (C) 2010 Elsevier Ltd. All rights reserved.206386392FAEPEX-UnicampConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

In search of a tolerance-induction strategy for cow's milk allergies: significant reduction of beta-lactoglobulin allergenicity via transglutaminase/cysteine polymerization

OBJECTIVE: To explore the use of beta-lactoglobulin polymerized using microbial transglutaminase and heating to identify whether protein polymerization could reduce in vivo allergenicity and maintain in vitro and ex vivo immunoreactivity for use in tolerance-induction protocols. METHODS: Based on previous protocols applied in mice and children, we performed in vivo challenges (using a skin prick test) with native and polymerized beta-lactoglobulin in adult patients with an IgE-mediated allergy to beta-lactoglobulin. In vitro humoral immunoreactivity was analyzed using immunoblotting. Cell-mediated immunoreactivity was analyzed using ex vivo challenges with native and polymerized beta-lactoglobulin and monitored by leukocyte adherence inhibition tests. RESULTS: The skin tests demonstrated that there was a significant reduction in immediate cutaneous reactivity after polymerization. Polymerization did not decrease the immunoblotting detection of s-IgE specific to beta-lactoglobulin. Cell-mediated immunoreactivity, as assessed by ex vivo challenges and leukocyte adherence inhibition tests, did not exhibit significant differences between leukocytes challenged with native versus polymerized beta-lactoglobulin. CONCLUSIONS: The polymerization of beta-lactoglobulin decreased in vivo allergenicity and did not decrease in vitro humoral or ex vivo cell-mediated immunoreactivity. Therefore, we conclude that inducing polymerization using transglutaminase represents a promising technique to produce suitable molecules for the purpose of designing oral/sublingual tolerance induction protocols for the treatment of allergies.67101171117