Ricardo Estrada

Clinical and laboratory alterations in horses during immunization with snake venoms for the production of polyvalent (Crotalinae) antivenom.

Six horses were immunized with the venoms of Bothrops asper, Crotalus durissus durissus and Lachesis muta stenophrys for the production of polyvalent (Crotalinae) antivenom. During the immunization, clinical and laboratory alterations were evaluated in these animals, and the development of humoral immune response was followed. Only moderate local tissue changes (edema, abscesses, fistules and fibrosis) were observed in these animals, whereas no systemic alterations occurred. Regarding laboratory tests, there was a drop in hemoglobin concentration and hematocrit, together with an increment in total serum protein. Horses developed a moderate leukocytosis, with increments in polymorphonuclear leukocytes and lymphocytes. No significant changes were observed in prothrombin time or platelet count. There were no alterations in serum lactic dehydrogenase and gamma glutamyl transferase activities, whereas minor increments in creatine kinase and alanine aminotransferase activities were observed, together with a decrease in aspartate aminotransferase. All these changes occurred after the injection of 9 mg venom, when sodium alginate was first used as adjuvant. Creatinine levels had a small increment, although no changes were observed in urea levels or in the urea/creatinine ratio. An important individual variability was observed in the humoral immune response, as judged not only by enzyme-linked immunosorbent assay, but also by assessing the neutralization of the indirect hemolytic activity of venoms.

On asymptotic expansions of twisted products

The series development of the quantum‐mechanical twisted product is studied. The series is shown to make sense as a moment asymptotic expansion of the integral formula for the twisted product, either pointwise or in the distributional sense depending on the nature of the factors. A condition is given that ensures convergence and is stronger than previously known results. Possible applications are examined.

A randomized double-blind clinical trial of two antivenoms in patients bitten by Bothrops atrox in Colombia

Abstract A randomized double-blind clinical trial in 39 patients envenomed by Bothrops atrox in Antioquia and Choco, Colombia, was performed to compare the efficacy and safety of 2 equine-derived antivenoms prepared at Instituto Clodomiro Picado, University of Costa Rica. Twenty patients received a monovalent anti- B , atrox antivenom (group A) and 19 patients were treated with a polyvalent (Crotalinae) antivenom (group B). Both antivenoms were equally efficient in the neutralization of the most relevant signs of envenoming (haemorrhage and blood clotting time alteration). Fourteen patients (36%) presented early adverse reactions to antivenoms and no significant difference between the 2 groups was observed. Urticaria (18%) was the most frequent early adverse reaction and there was no life-threatening anaphylactic reaction. Based on clinical criteria and serum venom levels, estimated by an enzyme immunoassay, 15 patients were classified into 2 groups: mild and moderate/severe envenoming. With the antivenom doses used in this study (3, 6 and 9 vials for mild, moderate and severe envenoming, respectively), both antivenoms were equally efficient in clearing serum venom levels within the first hour of treatment, and the levels remained below the lower limit of venom detection for 24 h. Antivenom concentration in serum remained high for up to 24 h after antivenom infusion, suggesting that an excess of antibody in relation to circulating antigen had been administered.

Immune response towards snake venoms.

The immune response involves a complex repertoire of innate and adaptive responses to foreign agents in the organism. The present review focuses on the immune response to snake venoms, including those occurring in snakebite accidental envenomation, experimental vaccination and animal hyperimmunization for snake antivenom production. The following aspects are considered: (a) the structural characteristics of snake toxins and their relationship to immunogenicity, (b) the effects that factors such as administration route, venom dose, type of adjuvant, and individual and species characteristics of the immunized animal have on the immune response, (c) the initial venom-induced inflammatory response, (d) the process by which specific antibodies towards individual toxins are produced, and (e) the techniques currently used to evaluate the antibody response. Understanding the immune response to snake venoms is highly relevant for improving antivenom production and for gaining a more complete view of snakebite envenoming.

Impact of Regional Variation in Bothrops asper Snake Venom on the Design of Antivenoms: Integrating Antivenomics and Neutralization Approaches

Intraspecific snake venom variations have implications in the preparation of venom pools for the generation of antivenoms. The impact of such variation in the cross-reactivity of antivenoms against Bothrops asper venom was assessed by comparing two commercial and four experimental antivenoms. All antivenoms showed similar immunorecognition pattern toward the venoms from adult and neonate specimens. They completely immunodepleted most P-III snake venom metalloproteinases (SVMPs), l-amino acid oxidases, serine proteinases, DC fragments, cysteine-rich secretory proteins (CRISPs), and C-type lectin-like proteins, and partially immunodepleted medium-sized disintegrins, phospholipases A(2) (PLA(2)s), some serine proteinases, and P-I SVMPs. Although all antivenoms abrogated the lethal, hemorrhagic, coagulant, proteinase, and PLA(2) venoms activities, monospecific experimental antivenoms were more effective than the polyspecific experimental antivenom. In addition, the commercial antivenoms, produced in horses subjected to repeated immunization cycles, showed higher neutralization than experimental polyspecific antivenom, produced by a single round of immunization. Overall, a conspicuous pattern of cross-neutralization was evident for all effects by all antivenoms, and monospecific antivenoms raised against venom from the Caribbean population were effective against venom from the Pacific population, indicating that geographic variations in venom proteomes of B. asper from Costa Rica do not result in overt variations in immunological cross-reactivity between antivenoms.

On Summability of Distributions and Spectral Geometry

Abstract:Modulo the moment asymptotic expansion, the Cesáro and parametric behaviours of distributions at infinity are equivalent. On the strength of this result, we construct the asymptotic analysis for spectral densities arising from elliptic pseudodifferential operators. We show how Cesáro developments lead to efficient calculations of the expansion coefficients of counting number functionals and Green functions. The bosonic action functional proposed by Chamseddine and Connes can more generally be validated as a Cesáro asymptotic development.

A distributional theory for asymptotic expansions

The purpose of this chapter is to present the distributional theory of asymptotic expansions for functions of one variable. This chapter and the next, where the multidimensional expansions are studied, are the central part of the book.

Regularization, pseudofunction, and hadamard finite part

Abstract First, we discuss and correlate the various types of regularizations available in the literature for the singular function H(x) x k , where k is an integer and H(x) is the Heaviside function. Then we present the corresponding regularization for the function r−k, where r is the radial distance in R n. Thereafter, we express the recently discovered distributional derivatives of this function in terms of pseudofunctional language commonly used in the Coulomb, gravitational, and interparticle potentials where the function 1 r plays a fundamental role.

Inhibition by CaNa2EDTA of local tissue damage induced by Bothrops asper (terciopelo) venom: Application in horse immunization for antivenom production

The ability of the chelating agent CaNa2EDTA to inhibit local tissue damage induced by Bothrops asper venom was studied in mice and in horses used for polyvalent (Crotalinae) antivenom production. CaNa2EDTA was devoid of toxicity when injected i.m. or s.c. inducing only a mild edema. Preincubation of B. asper venom with CaNa2EDTA inhibited hemorrhagic and dermonecrotic activities, but did not reduce edema-forming and myotoxic effects. A group of horses initially immunized with native venoms developed less severe local tissue reactions when injected with booster doses of venom and CaNa2EDTA than when receiving booster injections of venom alone, although they showed a similar antivenom response. Moreover, antivenoms produced from plasmas of horses that received booster injections of either venom alone or venom plus CaNa2EDTA had similar neutralizing activity against lethal, hemorrhagic and coagulant effects induced by B. asper venom. The similar antibody response was corroborated by Western blotting using crude venom and by an ELISA that estimates anti-myotoxin titer. It is concluded that the chelating agent CaNa2EDTA reduces the extent of local tissue damage induced by B. asper venom, without affecting the immune response of horses against pharmacologically-relevant venom components.

THE CESARO BEHAVIOUR OF DISTRIBUTIONS

A theory of summability for generalized functions of one real variable is presented; this theory generalizes the classical theory of summability of series and integrals. The relationship between the average behaviour of generalized functions according to this summability theory and the distributional behaviour is studied. Several applications are given.