Anaphylaxis induced by a drug containing lysozyme and papain: Influence of papain on the IgE response

[Background]: This paper reports the case of an egg-allergic pediatric patient who, once desensitized to egg following a successful rush oral immunotherapy protocol, could also tolerate Lizipaina®, a drug containing lysozyme (LYS) and papain, which had previously caused him a severe allergic reaction. Because the LYS amount that elicited the anaphylactic reaction (5 mg) was much lower than that tolerated during a double-blind placebo-controlled food challenge (corresponding to approximately 60 mg of LYS), the possibility that the presence of papain could increase the allergenic potential of LYS was investigated. [Methods]: Lizipaina, LYS and LYS hydrolyzed with papain were analyzed by SDS-PAGE under reducing and nonreducing conditions, and Western blotting of sera from egg-allergic patients was performed in order to detect IgE-binding fragments. Finally, sequence identification of the IgE-reactive bands was carried out by MALDI-TOF/TOF. [Results]: The SDS-PAGE pattern of LYS treated with papain under nonreducing conditions showed the presence of intact LYS that partially disappeared following reduction with β-mercaptoethanol, releasing IgE-reactive fragments as determined by Western blotting. MALDI-TOF/TOF revealed that papain degraded LYS, giving rise to three IgE-binding fragments: LYS (22-129), LYS (34-96) and LYS (62-128) that likely remained linked through the disulfide bonds present in the LYS molecule. Conclusion: The combined administration of LYS with proteolytic enzymes such as papain may have developed a severe allergic reaction in the patient studied, underlining the importance of considering all the components and their interactions when drugs are to be consumed by allergic persons.The authors acknowledge financial support from Fundación Salud 2000 (Merck-Serono, Spain) and ALK-Abelló Laboratories (Madrid, Spain), and by the project AGL2011-24740. S. B. and I.L.-E. acknowledge the financial support of CSIC through JAE-Pre and JAE-Doc grants, and D.L.-O. the financial support of the Ministerio de Educación, Cultura y Deporte through a FPU grant.Peer Reviewe

In vitro propagation and germplasm conservation of wild orchids from South America

Orchids are an important part of plant biodiversity on this planet due to their high variability among species and their habitats. South America represents more than thirty percent of all known orchid species, Colombia, Ecuador, Brazil, Peru, and Bolivia being among the richest countries in the world in terms of orchid biodiversity. Nevertheless, concerning the orchid conservation status, in Colombia precisely orchids occupy the unlucky first place as the plant family with the highest number of threatened species. There is a similar situation in the rest of the South American countries. The two main threats to orchid survival are both anthropogenic: the first one is deforestation, and the second largest threat to orchids is collection from the wild. One desirable action to safeguard these endangered species is to develop procedures that make possible their massive propagation, which would provide material for both environmental restoration and commercial purposes avoiding extractions from nature. Likewise, the development of systems that allow the ex situ conservation of orchid germplasm is imperative. This chapter reviews the progresses of different in vitro approaches for orchid propagation and germplasm conservation, safeguarding the genetic biodiversity of these species. Several study cases are presented and described to exemplify the protocols developed in the Botanical Institute of Northeast (UNNE-CONICET) for propagating and long-term storing the germplasm of wild orchids from Argentina (Cattleya lundii, Cohniella cepula, C. jonesiana, Gomesa bifolia, Aa achalensis, Cyrtopodium brandonianum, C. hatschbachii, Habenaria bractescens). Moreover, it has been attempted to put together most of the available literature on in vitro propagation and germplasm conservation for South American orchids using different explants and procedures. There are researches of good scientific quality that even cover critical insights into the physiology and factors affecting growth and development as well as storage of several orchid materials. Moreover, studies are still necessary to cover a major number of South American species as well as the use of selected material (clonal) for both propagation and conservation approaches.Fil: Dolce, Natalia Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; ArgentinaFil: Medina, Ricardo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; ArgentinaFil: Terada, Graciela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; ArgentinaFil: González Arnao, María Teresa. Universidad Veracruzana; MéxicoFil: Flachsland, Eduardo Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; Argentin