Los polímeros tipo elastina y su utilización como tags para la purificación de proteínas

Actualmente, una de las técnicas más ampliamente utilizadas en la purificación de proteínas recombinantes es la cromatografía de afinidad. Sin embargo, esta técnica es costosa, requiere de equipo especializado y es difícil de escalar. Por tanto, es deseable el desarrollo de métodos más económicos y técnicamente más sencillos. Uno de estos métodos está basado en aprovechar las características termosensibles y el comportamiento inteligente de los polímeros tipo elastina (ELP) para purificar una proteína de interés. El bajo coste que esta metodología requiere permitiría disminuir el precio de diversas proteínas de interés biomédico en el mercado, con las consiguientes repercusiones que ello conlleva a la hora de su aplicación en clínica. Por tanto, el presente artículo pretende indagar en la utilización de los ELP como tags para la purificación proteica, gracias al diseño y la producción de construcciones de fusión compuestas por la proteína diana de interés unida al tag elastomérico. Además se resaltarán otros efectos colaterales positivos que la presencia del ELP puede aportar a la proteína quimérica.Currently, chromatography is one of the most commonly used techniques to achieve protein purification. Nevertheless, such technique requires specialized equipment and is difficult to scale-up. Therefore, the development of new, simpler and broadly applicable purification methods to circumvent these problems is desirable. One such approach takes advantages of the thermo-sensitive and smart behavior of the elastin like polymers (ELP) to purify the target protein. The low-cost of carrying out such methodology may permit us to decrease the price of diverse biomedical useful proteins, with the consistent impact that such fact entails when applying in clinic. Therefore, the aim of this article is to clarify some aspects related to the use of the ELP as protein purification tags. For such use, it is necessary to design and produce fusion constructs between the target protein and the elastomeric tag. Moreover, apart from protein purification, further effects of the presence of the ELP in the fusion construct would be described.Peer ReviewedAward-winnin

Creation of antimicrobial biopolymers by the use of recombinant DNA technology

[Excerpt] The spread of antimicrobials resistant microorganisms has triggered the search for new ways to treat infections. One of these ways is the creation of antimicrobial devices and surfaces that kill or prevent the spread of microorganisms. In the present work we explored the properties of different antimicrobial peptides (AMPs) for the creation of biopolymers with broad antimicrobial activity. Antimicrobial recombinant protein-based polymers (rPBPs) were designed by cloning the DNA sequence coding for the different AMPs in frame with the N-terminus of the elastin-like recombinamer consisting of 200 repetitions of the pentamer VPAVG, here named A200. [...]This work was supported by the strategic programme UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569) funded by national funds through the FCT I.P. and by the ERDF through the COMPETE2020 - Programa Operacional Competitividade e Internacionalização (POCI). By the Spanish Minister of Economy and Competitiveness (MAT2012-38043-C02-01) and Junta de Castilla y León-JCyL (VA152A12-2 and VA155A12-2), Spain. AC and RM, acknowledge FCT for SFRH/BD/75882/2011 and SFRH/BPD/86470/2012 grants, respectively

Obtención de un polímero de tipo Elastina modificado con secuencias Bioactivas y Biodegradables, para su aplicación en ingeniería

La matriz extracelular es uno de los principales elementos reguladores de la actividad celular. Los diferentes módulos de las macromoléculas que la componen son capaces de desencadenar señales que activan diferentes rutas intracelulares que organizan las funciones vitales de las células. La ingeniería de tejidos se dedica a desarrollar sistemas capaces de imitar, temporalmente, el comportamiento de la matriz extracelular con objeto de promover la regeneración o el reemplazo de tejidos y órganos dañados, actuando como un soporte atractivo para las células que deben adherirse y crecer sobre ella, hasta reemplazarla por tejido sano. En este trabajo se describe el proceso de diseño y producción de un polímero de tipo elastina que se ha funcionalizado con secuencias bioactivas que añaden actividades específicas al andamio o soporte celular que constituye la elastina. Así,algunos dominios elastoméricos se modificaron con el aminoácido lisina para poder entrecruzar las moléculas de polímero y conseguir matrices. También se incluyó la secuencia REDV, presente el dominio CS5 de la fibronectina humana, como motivo de adhesión celular. Por último, el polímero se funcionalizó con secuencias diana de enzimas proteolíticas para mejorar su bioprocesabilidad.Extracellular matrix (ECM) is a major component for the regulation of cell activity. The different modules of the proteins which constitute the extracellular matrix macromolecules represent for the cells which enter in contact with them, new signals capable of activating several intracellular signaling pathways, resulting in the modulation of numerous cell functions. Tissue engineering tries to develop new materials based on these components as scaffolds for cells to promote their adhesion and growth. In this work, genetic engineering techniques were used to design and biosynthesize an extracellular matrix analogue based in the elastin component. The structural base of our scaffold is an elastin –derived sequence which confers an adequate mechanical behavior. In addition, several domains were included, for adding new bioactivities to this elastin-like polymer (ELP). Some of these elastic domains were modified to contain lysine for cross linking purposes. The polymer also contained periodically spaced fibronectin CS5 domain enclosing the well known cell attachment sequence REDV. Finally, the polymer had target sequences for proteolitic action.Peer ReviewedAward-winnin

Antibacterial protein-based fibres: combining recombinant DNA technology with electrospinning

[Excerpt] With the increasing healthcare-associated infections and antibiotic-resistant microorganisms there is a demand not only for new antimicrobial compounds but also for antimicrobial materials. Genetically engineered protein polymers functionalized with bioactive domains offer potential as multifunctional versatile materials for biomedical use. The present work describes the fabrication and characterization of antimicrobial fibre mats comprising the antimicrobial elastin-like recombinamer CM4-A200 [1]. [...]This work was supported by the strategic programme UID/BIA/04050/2013 (POCI-01- 0145-FEDER- 007569) through FCT I.P. and by ERDF through COMPETE2020 - POCI. The authors are grateful for funding from FCT (project “FunBioPlas” ERA-IB-2-6/0004/2014) and a fellowship to RM (SFRH-BPD/86470/2012). The authors also thank support from the COST Action MP1206

Commented checklist of marine fishes from the Galicia Bank seamount (NW Spain)

A commented checklist containing 139 species of marine fishes recorded at the Galician Bank seamount is presented. The list is based on nine prospecting and research surveys carried out from 1980 to 2011 with different fishing gears. The ichthyofauna list is diversified in 2 superclasses, 3 classes, 20 orders, 62 families and 113 genera. The largest family is Macrouridae, with 9 species, followed by Moridae, Stomiidae and Sternoptychidae with 7 species each. The trachichthyd Hoplostethus mediterraneus and the morid Lepidion lepidion were the most abundant species. Biogeographically, the Atlantic group, with 113 species (81.3%) is the best represented, followed by the Lusitanian one with 17 species (12.2%). Data on species abundance, as number of individuals caught, size and depth are reported. Habitat, distribution and vulnerability status are commented. Moreover, biometric data and meristic counts are also reported for several species. The results obtained showing a high fish biodiversity and a sensible number of threatened species, strongly support the future declaration of the Galicia Bank as a Marine Protected Area.En prensa0,974

Synthetic Protein Biotechnology approaches for the creation of antimicrobial biopolymers

[Excerpt] The spread of antimicrobials resistant microorganisms has triggered the search for new ways to treat infections. In the present work we explored the ABP-CM4 peptide properties from Bombyx mori for the creation of biopolymers with broad antimicrobial activity. An antimicrobial recombinant protein-based polymer (rPBP) was designed by cloning the DNA sequence coding for ABP-CM4 in frame with the N-terminus of the elastin-like recombinamer consisting of 200 repetitions of the pentamer VPAVG, here named A200. [...]This work was supported by FEDER through POFC – COMPETE by FCT through the project PEst-OE/BIA/UI4050/2014. By the Spanish Minister of Economy and Competitiveness (MAT2012-38043-C02-01) and Junta de Castilla y León-JCyL (VA152A12-2 and VA155A12-2), Spain. AC and RM, acknowledge FCT for SFRH/BD/75882/2011 and SFRH-BPD/86470/2012 grants, respectively

Antimicrobial Electrospun Membranes of Selp/Ag Composites

Silk-elastin-like proteins (SELPs) are a new class of bioinspired, biologically synthesized block copolymers, composed of silk and elastin repeating units. SELP electrospun fibre mats show potential for application as wound dressings for skin regeneration. In this work, antimicrobial nanofibrous mats were produced by electrospinning SELP solutions containing different concentrations of silver nitrate without addition of reducing agents. The SELP/Ag composite materials demonstrated antimicrobial activity against both Gram– and Gram+ bacteria. Furthermore, the SELP/Ag composite materials showed no cytotoxicity against normal human skin fibroblasts.This work was supported by FCT/MEC through Portuguese funds (PIDDAC) - PEst-OE/BIA/UI4050/2014, PEST-C/FIS/UI607/2011, Matepro - NORTE-07-0124-FEDER-000037. RM, AC and VS, acknowledge FCT for SFRH-BPD/86470/2012, SFRH/BD/75882/2011 and SFRH/BPD/63148/2009 grants, respectively. The authors also thank support from the COST Action MP1206 “Electrospun Nano-fibres for bio inspired composite materials and innovative industrial applications”

Non-motor symptom burden in patients with Parkinson's disease with impulse control disorders and compulsive behaviours : results from the COPPADIS cohort

The study was aimed at analysing the frequency of impulse control disorders (ICDs) and compulsive behaviours (CBs) in patients with Parkinson's disease (PD) and in control subjects (CS) as well as the relationship between ICDs/CBs and motor, nonmotor features and dopaminergic treatment in PD patients. Data came from COPPADIS-2015, an observational, descriptive, nationwide (Spain) study. We used the validated Questionnaire for Impulsive-Compulsive Disorders in Parkinson's Disease-Rating Scale (QUIP-RS) for ICD/CB screening. The association between demographic data and ICDs/CBs was analyzed in both groups. In PD, this relationship was evaluated using clinical features and treatment-related data. As result, 613 PD patients (mean age 62.47 ± 9.09 years, 59.87% men) and 179 CS (mean age 60.84 ± 8.33 years, 47.48% men) were included. ICDs and CBs were more frequent in PD (ICDs 12.7% vs. 1.6%, p < 0.001; CBs 7.18% vs. 1.67%, p = 0.01). PD patients had more frequent previous ICDs history, premorbid impulsive personality and antidepressant treatment (p < 0.05) compared with CS. In PD, patients with ICDs/CBs presented younger age at disease onset, more frequent history of previous ICDs and premorbid personality (p < 0.05), as well as higher comorbidity with nonmotor symptoms, including depression and poor quality of life. Treatment with dopamine agonists increased the risk of ICDs/CBs, being dose dependent (p < 0.05). As conclusions, ICDs and CBs were more frequent in patients with PD than in CS. More nonmotor symptoms were present in patients with PD who had ICDs/CBs compared with those without. Dopamine agonists have a prominent effect on ICDs/CBs, which could be influenced by dose

Estudios de marcado y recaptura de especies marinas

Los resultados obtenidos del marcado y posterior recaptura de los ejemplares son una herramienta muy valiosa para contribuir a mejorar el conocimiento de la biología y ecología de una especie, examinando ciertos aspectos como son: el crecimiento, los movimientos o migraciones, la mortalidad o supervivencia, la abundancia y distribución de la especie, el hábitat y diferenciación de poblaciones o stocks. Actualmente la técnica de marcado se aplica a muchas especies, tanto terrestres como marinas, pertenecientes a diversos grupos zoológicos: peces, crustáceos, reptiles, moluscos y mamíferos. Este libro repasa algunos ejemplos de marcado de especies marinas de interés comercial. No todas las especies pueden ser marcadas, porque es necesario cumplir una serie de requisitos para poder llevar a cabo con éxito un experimento de marcado. En uno de los apartados de esta guía, se describen los distintos aspectos a tener en cuenta para obtener buenos resultados. Se describen los principales proyectos de marcado actualmente en ejecución o en marcha llevados a cabo por el Instituto Español de Oceanografía (IEO). En primer lugar, se describe brevemente la especie, su distribución, crecimiento, reproducción, alimentación, etc. A continuación, se presenta la información del marcado, es decir, campañas realizadas, número de ejemplares marcados y algunos de los resultados obtenidos hasta la fecha a partir de las recapturas disponibles. En algunas especies, los programas de marcado se llevan realizando desde hace más de 20 años, como es el caso del atún rojo, por lo que la información disponible es bastante amplia. En otros casos por el contrario como la merluza, los proyectos son relativamente recientes, no obstante los resultados son bastante interesantes y prometedores.Nowadays many different marine animals are being tagged. This book summarizes recent tagging programs carried out by the Spanish Institute of Oceanography (IEO). Although the objectives of these various studies mainly depend on the species and each project in particular, the general aim is to better understand the biology and ecology of these animals the structure and dynamics of their populations and their capacity to respond to human activities. This book provides an overview of different aspects of this technique such as a brief history of tagging, the types of tags currently used, including both conventional and electronic tags, where and how to put them on the marine animals, some recommendations regarding how to perform a tagging survey and where to go or what to do if anyone recovers a tagged fish or marine animal. The book then summarizes the main species tagged by the IEO, making a short description of their biology followed by some of the results obtained from tagging studies undertaken until now. Other applications are to know the spatial distribution (spawning or feeding areas), estimate growth parameters, mortality and survival rates, longevity, the size of the population or identifying stocks. Nowadays the advances in electronics have also open new fields such us the possibility of tracking an animal and knowing its habitat preferences and behaviour. Besides some of these tags have the capacity of recording this information during long periods and sending the data from long distances even without the need to recover the animal. Tagging activities constitute a very useful tool to improve the knowledge of many species and contribute to their management and conservation. For that reason this methodology is included in many IEO projects in which other activities like the monitoring of the fishery (landings, fishing effort, fleet characteristics, fishing areas, biological sampling, etc.) are carried out. Some projects are related with coastal pelagic fisheries including anchovy, sardine and mackerel or oceanic pelagic fisheries like tuna and billfish species and pelagic sharks. Others are focused on benthic and demersal species such as hake, black spot seabream, anglerfish, flatfish, etc. Nevertheless not all species can be tagged, as they have to survive being caught and handled before being release. For this reason, tagging techniques may not easily be applied to some species.Versión del edito

RICORS2040 : The need for collaborative research in chronic kidney disease

Chronic kidney disease (CKD) is a silent and poorly known killer. The current concept of CKD is relatively young and uptake by the public, physicians and health authorities is not widespread. Physicians still confuse CKD with chronic kidney insufficiency or failure. For the wider public and health authorities, CKD evokes kidney replacement therapy (KRT). In Spain, the prevalence of KRT is 0.13%. Thus health authorities may consider CKD a non-issue: very few persons eventually need KRT and, for those in whom kidneys fail, the problem is 'solved' by dialysis or kidney transplantation. However, KRT is the tip of the iceberg in the burden of CKD. The main burden of CKD is accelerated ageing and premature death. The cut-off points for kidney function and kidney damage indexes that define CKD also mark an increased risk for all-cause premature death. CKD is the most prevalent risk factor for lethal coronavirus disease 2019 (COVID-19) and the factor that most increases the risk of death in COVID-19, after old age. Men and women undergoing KRT still have an annual mortality that is 10- to 100-fold higher than similar-age peers, and life expectancy is shortened by ~40 years for young persons on dialysis and by 15 years for young persons with a functioning kidney graft. CKD is expected to become the fifth greatest global cause of death by 2040 and the second greatest cause of death in Spain before the end of the century, a time when one in four Spaniards will have CKD. However, by 2022, CKD will become the only top-15 global predicted cause of death that is not supported by a dedicated well-funded Centres for Biomedical Research (CIBER) network structure in Spain. Realizing the underestimation of the CKD burden of disease by health authorities, the Decade of the Kidney initiative for 2020-2030 was launched by the American Association of Kidney Patients and the European Kidney Health Alliance. Leading Spanish kidney researchers grouped in the kidney collaborative research network Red de Investigación Renal have now applied for the Redes de Investigación Cooperativa Orientadas a Resultados en Salud (RICORS) call for collaborative research in Spain with the support of the Spanish Society of Nephrology, Federación Nacional de Asociaciones para la Lucha Contra las Enfermedades del Riñón and ONT: RICORS2040 aims to prevent the dire predictions for the global 2040 burden of CKD from becoming true