ABSTRACT
The COVID-19 pandemic has underscored the need for rapid and cost-effective diagnostic tools. Serological tests, particularly those measuring antibodies targeting the receptor-binding domain (RBD) of the virus, play a pivotal role in tracking infection dynamics and vaccine effectiveness. In this study, we aimed to develop a simple enzyme-linked immunosorbent assay (ELISA) for measuring RBD-specific antibodies, comparing two plant-based platforms for diagnostic reagent production. We chose to retain RBD in the endoplasmic reticulum (ER) to prevent potential immunoreactivity issues associated with plant-specific glycans. We produced ER-retained RBD in two plant systems: a stable transformation of BY-2 plant cell culture (BY2-RBD) and a transient transformation in Nicotiana benthamiana using the MagnICON system (NB-RBD). Both systems demonstrated their suitability, with varying yields and production timelines. The plant-made proteins revealed unexpected differences in N-glycan profiles, with BY2-RBD displaying oligo-mannosidic N-glycans and NB-RBD exhibiting a more complex glycan profile. This difference may be attributed to higher recombinant protein synthesis in the N. benthamiana system, potentially overloading the ER retention signal, causing some proteins to traffic to the Golgi apparatus. When used as diagnostic reagents in ELISA, BY2-RBD outperformed NB-RBD in terms of sensitivity, specificity, and correlation with a commercial kit. This discrepancy may be due to the distinct glycan profiles, as complex glycans on NB-RBD may impact immunoreactivity. In conclusion, our study highlights the potential of plant-based systems for rapid diagnostic reagent production during emergencies. However, transient expression systems, while offering shorter timelines, introduce higher heterogeneity in recombinant protein forms, necessitating careful consideration in serological test development.
ABSTRACT
Hairy root systems have proven to be a viable alternative for recombinant protein production. For recalcitrant proteins, maximizing the productivity of hairy root cultures is essential. The aim of this study was to optimize a Brassica rapa rapa hairy root process for secretion of alpha- l-iduronidase (IDUA), a biologic of medical value. The process was first optimized with hairy roots expressing eGFP. For the biomass optimization, the highest biomass yields were achieved in modified Gamborg B5 culture medium. For the secretion induction, the optimized secretion media was obtained with additives (1.5 g/l PVP + 1 mg/l 2,4- d + 20.5 g/l KNO3 ) resulting in 3.4 fold eGFP secretion when compared to the non-induced control. These optimized conditions were applied to the IDUA-expressing hairy root clone, confirming that the highest yields of secreted IDUA occurred when using the defined additive combination. The functionality of the IDUA protein, secreted and intracellular, was confirmed with an enzymatic activity assay. A > 150-fold increase of the IDUA activity was observed using an optimized secretion medium, compared with a non-induced medium. We have proven that our B. rapa rapa hairy root system can be harnessed to secrete recalcitrant proteins, illustrating the high potential of hairy roots in plant molecular farming.
Subject(s)
Biological Products , Brassica , Biological Products/metabolism , Brassica/genetics , Brassica/metabolism , Molecular Farming , Plant Roots/genetics , Plant Roots/metabolism , Recombinant Proteins/genetics , Recombinant Proteins/metabolismABSTRACT
Este artículo reflexiona sobre los supuestos de la investigación en salud basada en las artes (Art-Based Health Research) a partir de la experiencia del Proyecto Arte y Salud en el organismo público Madrid Salud del Ayuntamiento de Madrid. Las investigaciones en el ámbito del arte y la salud tienen una amplia trayectoria que permite un análisis crítico de los aportes y también una reflexión sobre su trascendencia e impacto. El artículo recoge aspectos de la investigación en salud basada en las artes, que se sustenta en la investigación basada en las artes y que plantean cuestiones y conflictos teóricos, metodológicos y éticos. El trabajo desarrollado en Madrid Salud desde en 2011 permite una revisión desde la práctica de estas cuestiones y un amplio campo de experimentación para reflexionar sobre los retos de este enfoque de investigación
This article reflects on the assumptions of Art-Based Health Research based on the experience of the Art and Health project in the public agency health of the Madrid City Council. Research in the field of art and health has a long trajectory that allows a critical analysis of the contributions and also a reflection on the transcendence and impact of such research. The article gathers aspects of Arts-Based Health Research, which is based on arts-based research, and which raise theoretical, methodological and ethical questions and conflicts. The work developed in Madrid's health since 2011 allows a review from the practice of these issues and a wide field of experimentation to reflect on the challenges of this research approach.
Subject(s)
Humans , Health Promotion , Health Programs and Plans , Health , Medicine in the ArtsABSTRACT
Hairy roots derived from the infection of a plant by Rhizobium rhizogenes (previously referred to as Agrobacterium rhizogenes) bacteria, can be obtained from a wide variety of plants and allow the production of highly diverse molecules. Hairy roots are able to produce and secrete complex active glycoproteins from a large spectrum of organisms. They are also adequate to express plant natural biosynthesis pathways required to produce specialized metabolites and can benefit from the new genetic tools available to facilitate an optimized production of tailor-made molecules. This adaptability has positioned hairy root platforms as major biotechnological tools. Researchers and industries have contributed to their advancement, which represents new alternatives from classical systems to produce complex molecules. Now these expression systems are ready to be used by different industries like pharmaceutical, cosmetics, and food sectors due to the development of fully controlled large-scale bioreactors. This review aims to describe the evolution of hairy root generation and culture methods and to highlight the possibilities offered by hairy roots in terms of feasibility and perspectives.
ABSTRACT
Este artículo presenta un resumen de lo que han sido los diez años de existencia del Proyecto curarte I+D, de la Universidad Complutense de Madrid y la Universidad de Salamanca. Objetivo: El objetivo prioritario del proyecto ha sido generar un modelo para integrar el arte en contextos de salud en España. Materiales y Métodos: En el artículo se analizan los referentes internacionales revisados y las fases de implementación del proyecto, desde sus inicios a partir de los primeros proyectos financiados, pasando por reuniones científicas y congresos, hasta la creación de programas académicos de máster y doctorado específicos. Resultados: Se presentan las principales, acciones logradas en estos años (humanización de espacios de salud, programas de talleres con niños y adolescentes hospitalizados, programas de arte para enfermos de Alzheimer y otras demencias, intervenciones artísticas, etc.). Conclusión: Todas estas acciones han situado al Proyecto curArte I+D como referente en España en este nuevo campo de estudio e investigación.
This article presents a summary of the ten years of existence of the curArte I+D Project created by Universidad Complutense de Madrid and Uninversidad de Salamanca.. Objective: The main objective of the project has been to develop, a model which integrates art in health contexts in Spain. Materials and Methods: In this article, international referents and implementation stges of the project are analyzed from the first sponsored projects, to the scientific meetings and congresses, to the generation of academic specific Masters and Doctoral programs. Results: The main actions achieved during these years (health spaces humanization, art programs for Alzheimer and other insanity patients, artistic nterventions, etc.) are presented. Conclusions: All these actions have placed the curArte Project I+D as a referent in this new field of study and research.
Este artigo apresenta um resumo do que tem sido os dez anos de existência do Projeto curArte I+D, da Universidade Complutense de Madrid e a Universidade de Salamanca. Objetivo: O objetivo prioritário do projeto tem sido gerar um modelo para integrar a arte em contextos de saúde em Espanha. Materiais e Métodos: No artigo se analisam os referentes internacionais revisados e as fases de aplicação do projeto, desde seus inícios a partir dos primeiros projetos financiados, passando por reuniões cientificas e congressos, até criação de programas acadêmicos de mestrado e doutorado específicos. Resultados: Se apresentam as principais, ações logradas nestes anos (humanização de espaços de saúde, programas de ensino com crianças e adolescentes hospitalizados, programas de arte para doentes de Alzheimer e outras demências, intervenções artísticas, etc.). Conclusões: Todas estas ações tem situado ao Projeto curArte I+D como referente em Espanha neste novo campo de estudo e pesquisa.
Subject(s)
Child , Art , Child Welfare , Education , Health , Quality of LifeABSTRACT
Después de caracterizar -desde la vertiente conceptual- los términos educación, pedagogía y maestría pedagógica se esbozan los componentes esenciales de la excelencia pedagógica en la educación médica superior, así como los requisitos indispensables que debe reunir el profesor de Ciencias Médicas para alcanzar esa categoria excepcional en la docencia médica superior
Subject(s)
Education, MedicalABSTRACT
Después de caracterizar -desde la vertiente conceptual- los términos educación, pedagogía y maestría pedagógica se esbozan los componentes esenciales de la excelencia pedagógica en la educación médica superior, así como los requisitos indispensables que debe reunir el profesor de Ciencias Médicas para alcanzar esa categoria excepcional en la docencia médica superior(AU)
Subject(s)
Education, MedicalABSTRACT
En el estudio prospectivo de 20 reuniones administrativas en una comunidad terapéutica mixta de 50 pacientes en hospitalización total, basado en el registro y calificación de las incidencias de cada reunión efectuada, y las respuestas a encuestas anónimas aplicadas a 200 egresados se obtuvieron resultados que evidenciaron el desarrollo positivo de la actividad, la participación del colectivo en las gestiones comunitarias y la elevada significación de dicha reunión para los pacientes que la calificaron como estimulante y facilitadora de interacciones personales. Los autores concluyen que la inclusión de pacientes alcohólicos en proporción adecuada favorecía la cohesión intragrupo(AU)
