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

Cell Lineage Tracing Identifies Hormone-Regulated and Wnt-Responsive Vaginal Epithelial Stem Cells.

The intact vaginal epithelium is essential for women's reproductive health and provides protection against HIV and sexually transmitted infections. How this epithelium maintains itself remains poorly understood. Here, we used single-cell RNA sequencing (RNA-seq) to define the diverse cell populations in the vaginal epithelium. We show that vaginal epithelial cell proliferation is limited to the basal compartment without any obvious label-retaining cells. Furthermore, we developed vaginal organoids and show that the basal cells have increased organoid forming efficiency. Importantly, Axin2 marks a self-renewing subpopulation of basal cells that gives rise to differentiated cells over time. These cells are ovariectomy-resistant stem cells as they proliferate even in the absence of hormones. Upon hormone supplementation, these cells expand and reconstitute the entire vaginal epithelium. Wnt/β-catenin is essential for the proliferation and differentiation of vaginal stem cells. Together, these data define heterogeneity in vaginal epithelium and identify vaginal epithelial stem cells

Development of a mechanism and an accurate and simple mathematical model for the description of drug release: Application to a relevant example of acetazolamide-controlled release from a bio-inspired elastin-based hydrogel

Producción CientíficaTransversality between mathematical modeling, pharmacology, and materials science is essential in order to achieve controlled-release systems with advanced properties. In this regard, the area of biomaterials provides a platform for the development of depots that are able to achieve controlled release of a drug, whereas pharmacology strives to find new therapeutic molecules and mathematical models have a connecting function, providing a rational understanding by modeling the parameters that influence the release observed. Herein we present a mechanism which, based on reasonable assumptions, explains the experimental data obtained very well. In addition, we have developed a simple and accurate “lumped” kinetics model to correctly fit the experimentally observed drug release behavior. This lumped model allows us to have simple analytic solutions for the mass and rate of drug release as a function of time without limitations of time or mass of drug released, which represents an important step-forward in the area of in vitro drug delivery when compared to the current state of the art in mathematical modeling. As an example, we applied the mechanism and model to the release data for acetazolamide from a recombinant polymer. Both materials were selected because of a need to develop a suitable ophthalmic formulation for the treatment of glaucoma. The in vitro release model proposed herein provides a valuable predictive tool for ensuring product performance and batch-to-batch reproducibility, thus paving the way for the development of further pharmaceutical devices.Este trabajo forma parte de los Proyectos de Investigación financiados por la Comisión Europea a través del Fondo Social Europeo (FSE) y de la Consejería de Educación mediante el Fondo Europeo de Desarrollo Regional (ERDF), el MINECO (Proyectos MAT2013-41723-R, MAT2013-42473-R, PRI−PIBAR-2011-1403 y MAT2012-38043), la Junta de Castilla y León (Proyectos VA155A12, VA152A12, and VA244U13), el CIBER-BBN y el Instituto de Salud Carlos III mediante el Centro de Medicina Regenerativa y Terapia Celular de Castilla y León

Flux flow resistivity and vortex viscosity of high-Tc films

The flux flow regime of high-T$_{\rm c}$ samples of different normal state resistivities is studied in the temperature range where the sign of the Hall effect is reversed. The scaling of the vortex viscosity with normal state resistivity is consistent with the Bardeen-Stephen theory. Estimates of the influence of possible mechanisms suggested for the sign reversal of the Hall effect are also given.Comment: 3 pages. 4 figures upon reques

Amphiphilic Elastin-Like Block Co-Recombinamers Containing 2 Leucine Zippers: Cooperative Interplay between Both Domains 3 Results in Injectable and Stable Hydrogels

Many biological processes are regulated by reversible binding events, with these interactions between macromolecules representing the core of dynamic chemistry. As such, any attempt to gain a better understanding of such interactions, which would pave the way to the extrapolation of natural designs to create new advanced systems, is clearly of interest. This work focuses on the development of a leucine zipper-elastin-like recombinamer (ZELR) in order to elucidate the behavior of such domains when coexisting along the same molecule and to engineer reversible, injectable and stable hydrogels. The unique propensity of the Z-moiety selected to dimerize, together with the thermosensitive behavior of the ELR, which has been constructed as a thermosensitive amphiphilic tetrablock, has been engineered into a single recombinant molecule. In this molecular design, the Z-moieties are unable to form a network, while the ELR is below its Tt, thus, guaranteeing the liquid-like state of the system. However, this situation changes rapidly as the temperature increases above Tt, where a stable hydrogel is formed, as demostrated by rheological tests. The inability of the ELR molecule (without Z-domains) to form such a stable hydrogel above Tt clearly points to a positive cooperative effect between these two domains (Z and EL), and no conformational changes in the former are involved, as demonstrated by circular dichroism analysis. AFM shows that Z-motifs seem to induce the aggregation of micelles, which supports the enhanced stability displayed by ZELRs when compared to ELR at the macroscale level. To the best of our knowledge, this is the first time that such an interplay between these two domains has been reported. Furthermore, the cytocompatibility of the resulting hydrogels opens the door to their use in biomedical applications.Este trabajo forma parte de Proyectos de Investigación financiados por la Comisión Europea a través del Fondo Social Europeo (FSE) y el Fondo Europeo de Desarrollo Regional (ERDF), por el del MINECO (MAT2013-41723R, MAT2013-42473-R, MAT2012-38043 y PRI-PIBAR-2011-1403), la Junta de Castilla y León (VA049A11, VA152A12 y VA155A12) y el Instituto de Salud Carlos III bajo el Centro en Red de Medicina Regenerativa y Terapia Celular de Castilla y León

Heat inactivated mycobacteria, alpha-Gal and zebrafish: Insights gained from experiences with two promising trained immunity inductors and a validated animal model

Trained immunity (TRAIM) may be defined as a form of memory where innate immune cells such as monocytes, macrophages, dendritic and natural killer (NK) cells undergo an epigenetic reprogramming that enhances their primary defensive capabilities. Cross-pathogen protective TRAIM can be triggered in different hosts by exposure to live microbes or microbe-derived products such as heat-inactivated Mycobacterium bovis or with the glycan α-Gal to elicit protective responses against several pathogens. We review the TRAIM paradigm using two models representing distinct scales of immune sensitization: the whole bacterial cell and one of its building blocks, the polysaccharides or glycans. Observations point out to macrophage lytic capabilities and cytokine regulation as two key components in non-specific innate immune responses against infections. The study of the TRAIM response deserves attention to better characterize the evolution of host-pathogen cooperation both for identifying the aetiology of some diseases and for finding new therapeutic strategies. In this field, the zebrafish provides a convenient and complete biological system that could help to deepen in the knowledge of TRAIM-mediated mechanisms in pathogen-host interactions.This study was funded by Junta de Comunidades de Castilla-La Mancha, Spain, and EU-FEDER, projects MYCOTRAINING SBPLY/19/180501/000174 and CCM17-PIC-036 (SBPLY/17/180501/000185), Ministerio de Ciencia e Innovación/Agencia Estatal de Investigación MCIN/AEI/10.13039/501100011033, Spain and EU-FEDER (Grant BIOGAL PID2020-116761GB-I00) and by Principado de Asturias, PCTI 2021–2023 (GRUPIN: IDI2021-000102) and FEDER. EFC holds a PhD contract from the UCLM cosupported by the European Social Fund (2020/3836).S

Experimental and Computational Fluid Dynamic study of an active ventilated façade integrating battery and distributed MPPT

Ventilated Façades integrating photovoltaic panels are a promising way to improve efficiency and the thermal-physical performances of buildings. Due the inherent intermittence of the non-programmable renewable energy sources, their increasing usage implies the use of energy storage systems to mitigate the mismatch between power generation and the buildings’ load demand. The main purpose of this paper is to investigate the thermo-fluid dynamic performances of a prototype integrating a photovoltaic cell and a battery as a module of an active ventilated façade. Based on an experimental setup, a numerical study in steady state conditions of flow through the air cavity of the module has been carried out and implemented in a fluid-dynamics Finite Volume code. In order to assess the viability of the prototype, the calibrated model was lastly used to predict thermal performance of the prototype on different climate conditions supporting its further improvement

Raiva em herbívoros no estado do Pará, Brasil: estudo descritivo (2004 a 2013)

PARC/PROPESP and PAPQ/ PROPESPUniversidade Federal do Pará. Instituto de Medicina Veterinária. Laboratório de Epidemiologia e Geoprocessamento. Castanhal, PA, Brazil.Instituto Federal de Educação do Tocantins. Palmas, TO, Brazil.Universidade Federal do Pará. Instituto de Medicina Veterinária. Laboratório de Epidemiologia e Geoprocessamento. Castanhal, PA, Brazil.Universidade Federal de Mato Grosso. Graduate Program in Health Sciences. Sinop, MT, Brazil.Museu Paraense Emílio Goeldi - Campus de Pesquisa. Programa de Capacitação Institucional. Coordenação Ciências da Terra e Ecologia. Belém, PA, Brazil.Agência de Defesa Agropecuária do Pará. Belém, PA, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Geoprocessamento. Ananindeua, PA, Brasil.Universidade Federal do Pará. Instituto de Medicina Veterinária. Laboratório de Epidemiologia e Geoprocessamento. Castanhal, PA, Brazil.Rabies is an important zoonosis to public health associated with lethal encephalitis and economic losses. Analysis of its spatial distribution is a meaningful tool in understanding its dispersion, which may contribute to the control and prophylaxis of the disease. This study analyzed the spatial-temporal distribution of rabies outbreaks in livestock in Pará state, Brazil, from 2004 to 2013. We used records of neurological syndromes obtained from the state’s livestock authority (Adepará). The analysis recorded 711 neurological syndromes reports in livestock, of which 32.8% were positive for rabies. In 8% of the neurological syndromes (n=57) was not possible to perform the analysis because of bad-packaging conditions of the samples sent. Outbreaks involved at least 1,179 animals and cattle were the most affected animal species (76.8%). The numbers of reported neurological syndromes and of rabies outbreak shad strong positive correlation and exhibited decreasing linear trend. Spatially, most outbreaks occurred in two mesoregions in Pará (Northeast and Southeast). One of the justifications for this spatial distribution may be related with the distribution of the animals in the state, since these mesoregions are the largest cattle producers in Pará and have most of their territory deforested for pasture implementation

Ferromagnetic supramolecular metal-organic frameworks for active capture and magnetic sensing of emerging drug pollutants

Capture and sensing of emerging pollutants is one of the increasing environmental concerns due to the adverse ecological and human health effects. Here, we report the synthesis of a supramolecular metal-organic framework (SMOF) [CrCu6(m-H2O)6(m3-OH)6(m-adeninato-kN33:kN9)6](SO4)1.5 which is able to capture anionic drugs and exhibits magnetic properties useful for sensing purposes. The features of the nucleobase decorated CrCu6 building block allow the incorporation of up to 9 drug molecules (i.e., ibuprofen and naproxen in this work) per heptameric entity. In addition, we provide a simple way to quantify the incorporated number of drug molecules through a magnetic sustentation experiment in which the field required to keep the particles attached to the electromagnet pole is linearly related to the total mass of the anionic counterion. In this way, it also provides an easy way to determine the amount of entrapped drug molecules, making this SMOF a promising candidate for environmental remediation technologies.This work has been funded by the Universidad del País Vasco/Euskal Herriko Unibertsitatea (GIU17/50), the Gobierno Vasco/Eusko Jaurlaritza (PIBA18/14; IT1291-19), the Ministerio de Economía y Competitividad (MAT2016-75883-C2-1-P), the Ministerio de Ciencia e Innovación (PID2019-108028GB-C21), and FEDER funds. Technical and human support provided by SGIker (UPV/EHU, MICINN, GV/EJ, and ESF) is also acknowledged