IL-10 deficiency exacerbates the brain inflammatory response to permanent ischemia without preventing resolution of the lesion

El pdf del artículo es la versión post-print.Stroke induces inflammation that can aggravate brain damage. This work examines whether interleukin-10 (IL-10) deficiency exacerbates inflammation and worsens the outcome of permanent middle cerebral artery occlusion (pMCAO). Expression of IL-10 and IL-10 receptor (IL-10R) increased after ischemia. From day 4, reactive astrocytes showed strong IL-10R immunoreactivity. Interleukin-10 knockout (IL-10 KO) mice kept in conventional housing showed more mortality after pMCAO than the wild type (WT). This effect was associated with the presence of signs of colitis in the IL-10 KO mice, suggesting that ongoing systemic inflammation was a confounding factor. In a pathogen-free environment, IL-10 deficiency slightly increased infarct volume and neurologic deficits. Induction of proinflammatory molecules in the IL-10 KO brain was similar to that in the WT 6 hours after ischemia, but was higher at day 4, while differences decreased at day 7. Deficiency of IL-10 promoted the presence of more mature phagocytic cells in the ischemic tissue, and enhanced the expression of M2 markers and the T-cell inhibitory molecule CTLA-4. These findings agree with a role of IL-10 in attenuating local inflammatory reactions, but do not support an essential function of IL-10 in lesion resolution. Upregulation of alternative immunosuppressive molecules after brain ischemia can compensate, at least in part, the absence of IL-10. © 2013 ISCBFM.Work supported by the Spanish Ministry of Economy (SAF2011-30492), and the European Community (FP7, grant agreements: n°201024 ARISE and n°278850 InMiND), and the ERANET-NEURON project (PRI-PIMNEU-2011-1342). IPP and EBT had PhD fellowships from the Agència de Gestió d'Ajuts Universitaris i de Recerca (AGAUR) of the Generalitat de Catalunya and the FPU program of the Spanish Ministry of Economy, respectively.Peer Reviewe

Controlled manipulation of enzyme specificity through immobilization-induced flexibility constraints

It is thought that during immobilization enzymes, as dynamic biomolecules, may become distorted and this may alter their catalytic properties. However, the effects of different immobilization strategies on enzyme rigidity or flexibility and their consequences in specificity and stereochemistry at large scale has not been yet clearly evaluated and understood. This was here investigated by using as model an ester hydrolase, isolated from a bacterium inhabiting a karstic lake, with broad substrate spectrum (72 esters being converted; 61.5 U mg−1 for glyceryl tripropionate) but initially non-enantiospecific. We found that the enzyme (7 nm × 4.4 nm × 4.2 nm) could be efficiently ionic exchanged inside the pores (9.3 nm under dry conditions) of amino-functionalized ordered mesoporous material (NH2-SBA-15), achieving a protein load of 48 mg g−1, and a specific activity of 4.5 ± 0.1 U mg−1. When the enzyme was site-directed immobilized through His interaction with an immobilized cationon the surface of two types of magnetic micro-particles through hexahistidine-tags, protein loads up to 10.2 μg g−1 and specific activities of up to 29.9 ± 0.3 U mg−1, were obtained. We found that ionically exchanged enzyme inside pores of NH2-SBA-15 drastically narrowed the substrate range (17 esters), to an extent much higher than ionically exchanged enzyme on the surface of magnetic micro-particles (up to 61 esters). This is attributed to differences in surface chemistry, particle size, and substrate accessibility to the active site tunnel. Our results also suggested, for the first time, that immobilization of enzymes in pores of similar size may alter the enzyme structures and produce enzyme active centers with different configuration which promote stereochemical conversions in a manner different to those arising from surface immobilization, where the strength of the ionic exchange also has an influence. This was shown by demonstrating that when the enzyme was introduced inside pores with a diameter (under dry conditions) slightly higher than that of the enzyme crystal structure a biocatalyst enantiospecific for ethyl (R)-4-chloro-3-hydroxybutyrate was produced, a feature not found when using wider pores. By contrast, immobilization on the surface of ferromagnetic microparticles produced selective biocatalysts for methyl (S)-(+)-mandelate or methyl (S)-lactate depending on the functionalization. This study illustrates the benefits of extensive analysis of the substrate spectra to better understand the effects of different immobilization strategies on enzyme flexibility/rigidity, as well as substrate specificity and stereochemistry. Our results will help to design tunable materials and interfaces for a controlled manipulation of specificity and to transform non-enantiospecific enzymes into stereo-chemically substrate promiscuous biocatalysts capable of converting multiple chiral molecules.This project received funding from the European Union’s Horizon 2020 research and innovation program Blue Growth: Unlockingthe potential of Seas and Oceans under grant agreement no. 634486 (project acronym INMARE). This research was also supported by the grants PCIN-2014-107 (within ERA NET IB2 grant nr. ERA-IB-14-030 - MetaCat), PCIN-2017-078 (within the ERA-MarineBiotech grant ProBone), BIO2014-54494-R, MAT2016-77496-R, BIO2017-85522-R, and CTQ2016-79138-R from the Spanish Ministry of Economy, Industry and Competitiveness. A.B. acknowledges the support from the Spanish Ministry of Economy, Industry and Competitiveness (MAT2017-88808-R grant), María de Maeztu Units of Excellence Programme (MDM-2016-0618), and the Diputación de Guipúzcoa for current funding in the frame of Gipuzkoa Fellows program. G.D. thanks the German Federal Ministry of Education and Research (BMBF, Grant No. 031A095C) for funding in the frame of the Molecular Interaction Engineering program (Biotechnologie 2020+). The authors gratefully acknowledge financial support provided by the European Regional Development Fund (ERDF). C.C. thanks the Spanish Ministry of Economy, Industry and Competitiveness for a PhD fellowship (Grant BES-2015-073829).Peer ReviewedPostprint (published version

Controlled manipulation of enzyme specificity through immobilization-induced flexibility constraints

It is thought that during immobilization enzymes, as dynamic biomolecules, may become distorted and this may alter their catalytic properties. However, the effects of different immobilization strategies on enzyme rigidity or flexibility and their consequences in specificity and stereochemistry at large scale has not been yet clearly evaluated and understood. This was here investigated by using as model an ester hydrolase, isolated from a bacterium inhabiting a karstic lake, with broad substrate spectrum (72 esters being converted; 61.5 U mg$^{-1}$ for glyceryl tripropionate) but initially non-enantiospecific. We found that the enzyme (7 nm × 4.4 nm × 4.2 nm) could be efficiently ionic exchanged inside the pores (9.3 nm under dry conditions) of amino-functionalized ordered mesoporous material (NH$_{2}$-SBA-15), achieving a protein load of 48 mg g−1, and a specific activity of 4.5 ± 0.1 U mg$^{-1}$. When the enzyme was site-directed immobilized through His interaction with an immobilized cationon the surface of two types of magnetic micro-particles through hexahistidine-tags, protein loads up to 10.2 μg g$^{-1}$ and specific activities of up to 29.9 ± 0.3 U mg$^{-1}$, were obtained. We found that ionically exchanged enzyme inside pores of NH2-SBA-15 drastically narrowed the substrate range (17 esters), to an extent much higher than ionically exchanged enzyme on the surface of magnetic micro-particles (up to 61 esters). This is attributed to differences in surface chemistry, particle size, and substrate accessibility to the active site tunnel. Our results also suggested, for the first time, that immobilization of enzymes in pores of similar size may alter the enzyme structures and produce enzyme active centers with different configuration which promote stereochemical conversions in a manner different to those arising from surface immobilization, where the strength of the ionic exchange also has an influence. This was shown by demonstrating that when the enzyme was introduced inside pores with a diameter (under dry conditions) slightly higher than that of the enzyme crystal structure a biocatalyst enantiospecific for ethyl (R)-4-chloro-3-hydroxybutyrate was produced, a feature not found when using wider pores. By contrast, immobilization on the surface of ferromagnetic microparticles produced selective biocatalysts for methyl (S)-(+)-mandelate or methyl (S)-lactate depending on the functionalization. This study illustrates the benefits of extensive analysis of the substrate spectra to better understand the effects of different immobilization strategies on enzyme flexibility/rigidity, as well as substrate specificity and stereochemistry. Our results will help to design tunable materials and interfaces for a controlled manipulation of specificity and to transform non-enantiospecific enzymes into stereo-chemically substrate promiscuous biocatalysts capable of converting multiple chiral molecules

Unexpected discovery: “A new 3,3′-bipyrazolo[3,4-b]pyridine scaffold and its comprehensive analysis”

In this study, a novel 3,3′-bipyrazolo [3,4-b]pyridine-type structure was synthesized from 5-acetylamino-3-methyl-1-phenylpyrazole using the Vilsmeier-Haack reaction as a key step. The spectroscopic properties and structural elucidation of the compound were determined with the use of FT-IR, HRMS, 1H NMR, and 13C NMR. Likewise, the theoretical analysis of the IR and NMR spectra allowed peaks to be assigned and a solid correlation was demonstrated between the experimental and theoretical results. Finally, ab initio calculations based on the density functional theory method at the B3LYP/6-311G (d,p) level of theory were used to determine the conformational energy barrier, facilitating the identification of the most probable conformers of the synthesized compound. Overall, our findings contribute to the understanding of bipyrazolo [3,4-b]pyridine derivatives.Fondo Nacional de Desarrollo Científico y Tecnológico (Chile)9 página

Biology of moderately halophilic aerobic bacteria

The moderately halophilic heterotrophic aerobic bacteria form a diverse group of microorganisms. The property of halophilism is widespread within the bacterial domain. Bacterial halophiles are abundant in environments such as salt lakes, saline soils, and salted food products. Most species keep their intracellular ionic concentrations at low levels while synthesizing or accumulating organic solutes to provide osmotic equilibrium of the cytoplasm with the surrounding medium. Complex mechanisms of adjustment of the intracellular environments and the properties of the cytoplasmic membrane enable rapid adaptation to changes in the salt concentration of the environment. Approaches to the study of genetic processes have recently been developed for several moderate halophiles, opening the way toward an understanding of haloadaptation at the molecular level. The new information obtained is also expected to contribute to the development of novel biotechnological uses for these organisms

Certificación turística en América: Análisis descriptivo hasta el 2023

La certificación en el turismo puede servir tanto de estímulo para mejorar la gestión de los recursos turísticos como de incentivo para los visitantes, puesto que a menudo ofrecen garantía de buenas experiencias turísticas. En tal sentido, es necesario realizar investigaciones que muestren las características de los sistemas de certificación turística en los destinos. El objetivo de este artículo es describir los sistemas de certificación turísticas en las Américas. Para el estudio se consideró un total de 59 sistemas de certificación turística en el continente, con niveles operativos globales, subglobales, nacionales, regionales y locales. La metodología se sustentó en una investigación de tipo documental, caracterización cualitativa y cuantitativa de los sistemas de certificación, utilizando la estadística descriptiva y el programa SPSS para el procesamiento de los datos. Entre los resultados se obtuvo que la mayor categoría certificada en el continente es “Proveedores de alojamiento”; “Estados Unidos” posee el mayor número de programas de certificación; el nivel operativo predominante entre los sistemas de certificación es el “Regional”. Se concluye que, muchos países de América utilizan los mismos criterios de sistemas globales para construir sus programas de certificación turística nacionales, en línea con las políticas ambientales y turísticas propias de cada país

Calidad microbiológica de una galleta formulada a base de harina de yuca y plasma de bovino

En este trabajo se evaluó la calidad microbiológica de una galleta formulada con harina de yuca y plasma de bovino. Las galletas una vez horneadas fueron enfriadas a temperatura ambiente durante 1 h, para luego ser empacadas en bolsas de celofán cerradas herméticamente. Posteriormente, se almacenaron en condiciones de despensa (25ºC-28ºC) y rotuladas con los respectivos días de almacenamiento. Al mencionado producto se le realizó, por medio de la técnica Petrifilm 3M, la determinación de aerobios mesófilos, coliformes, Escherichia coli, mohos y levaduras, mientras que al plasma de bovino (utilizado como materia prima), además de los contajes microbiológicos arriba mencionados, se determinaron aerobios mesófilos, coliformes, Escherichia coli y la presencia de Staphylococcus aureus y Salmonella. El análisis microbiológico del producto, fue realizado a los días 0, 7, 10, 13, 15 y 20, mientras que para el plasma de bovino, el análisis se llevó a cabo el mismo día de su obtención bajo condiciones de refrigeración. Los resultados obtenidos revelaron que en los primeros 15 días, el recuento total de aerobios (RTA) se encontró dentro de los límites permitidos por la norma COVENIN, no obstante; a partir del día 20 el producto mostró un crecimiento de éste por encima de lo permitido en las mencionadas normas. El resto de los microorganismos estudiados estuvieron ausentes. Los resultados demuestran que la galleta presentó buenas características microbiológicas durante un período de 15 días de almacenamiento, no detectándose microorganismos de interés sanitario, aún cuando la misma fue almacenada a temperatura de despensa sin ningún tipo de preservativos

Influence of different bottom types on the Senegal sole Solea senegalensis Kaup, 1858 ongrowing

Fish coloration may affect its market value. The present paper compares growth, survival rate and pigmentation during the ongrowing of Senegal sole Solea senegalensis Kaup, 1858 on three different bottoms. This experiment lasted five months, and at the beginning fish were 4.93 ± 0.75 cm (length) and 1.91 ± 0.68 g (weight). No significant differences were found for final length and weight between different bottoms. Several pigmentation patterns were found on the sand and white bottoms, but one single pattern appeared in the dark bottom tanks. The survival rate on the sand bottom was significantly less than others, probably due to pathologies stemming from the difficult maintenance of the sand bed.La falta de homogeneidad en la coloración de los peces puede afectar a su valor comercial. En este trabajo se ha comparado el crecimiento, la supervivencia y la pigmentación de ejemplares de lenguado senegalés Solea senegalensis Kaup, 1858 durante el preengorde en tanques con tres tipos de fondos. El experimento tuvo una duración de 5 meses. El tamaño inicial de los individuos era 4,93 ± 0,75 cm y 1,91 ± 0,68 g, y al final de la experiencia no se encontraron diferencias significativas entre las tallas y los pesos de los ejemplares de los distintos tanques. Se detectaron varios patrones de pigmentación en los especímenes correspondientes a los fondos arenoso y blanco, pero solo uno en los de los tanques de fondo gris. La supervivencia en el fondo de arena fue la menor de todas (p 0,01), probablemente atribuible a patologías asociadas al difícil mantenimiento del lecho arenoso.Instituto Español de Oceanografí

White dwarf-main-sequence binaries from Gaia EDR3: The unresolved 100 pc volume-limited sample

We use the data provided by the Gaia Early Data Release 3 to search for a highly-complete volume-limited sample of unresolved binaries consisting of a white dwarf and a main sequence companion (i.e. WDMS binaries) within 100 pc. We select 112 objects based on their location within the Hertzsprung-Russell diagram, of which 97 are new identifications. We fit their spectral energy distributions (SED) with a two-body fitting algorithm implemented in VOSA (Virtual Observatory SED Analyser) to derive the effective temperatures, luminosities and radii (hence surface gravities and masses) of both componentsARM acknowledges financial support from the MINECO under the Ramón y Cajal program (RYC-2016-20254). ST and ARM acknowledge support from the MINECO under the AYA2017-86274-P grant, and the AGAUR grant SGR-661/2017. ESM and FJE acknowledge financial support from the MINECO under the AYA2017-86274-P grant. FJE acknowledges support from the H2020 ESCAPE project (Grant Agreement no. 824064). LMC, LGA and AHC acknowledge support from AGENCIA through the Programa de Modernización Tecnológica BID 1728/OC-AR, and from CONICET through the PIP 2017-2019 GI grant. This publication makes use of VOSA and SVO DiscTool, developed under the Spanish Virtual Observatory project supported from the Spanish MINECO through grant AyA2017-84089. This research has made use of Aladin sky atlas developed at CDS, Strasbourg Observatory, France (Bonnarel et al. 2000; Boch & Fernique 2014). TOPCAT (Taylor 2005) and STILTS (Taylor 2006) have also been widely used in this paper. We thank the anonymous referee for the helpful suggestions. The authors are greatly indebted to Detlev Koester for sharing his grid of model atmosphere white dwarf spectra. The authors also thank Roberto Raddi for sharing the grid of white dwarf absolute magnitudes calculated for the Gaia EDR3 bandpasses. This work has made use of data from the European Space Agency (ESA) mission Gaia (https://www.cosmos.esa.int/ gaia), processed by the Gaia Data Processing and Analysis Consortium (DPAC, https://www.cosmos.esa.int/web/gaia/dpac/ consortium). Funding for the DPAC has been provided by national institutions, in particular the institutions participating in the Gaia Multilateral AgreementPostprint (updated version

Analysis and description of disease-specific quality of life in patients with anal fistula

INTRODUCTION: In patients diagnosed with anal fistula, knowing the quality of life specifically related to the disease can help coloproctology specialists to choose the most appropriate therapeutic strategy for each case. The aim of our study is to analyzse and describe the factors related to the specific quality of life in a consecutive series of patients diagnosed with anal fistula. METHODS: Observational, cross-sectional study carried out from March 2015 to February 2017. All patients were assessed in the colorectal surgery unit of a hospital in southeast of Spain. After performing an initial anamnesis and a physical examination, patients diagnosed with anal fistula completed the Quality of Life in Ppatients with Anal Fistula Questionnaire (QoLAF-Q). This questionnaire specifically measures quality of life in people with anal fistula and its score range is the following: zero impact = 14 points, limited impact = 15 to 28 points, moderate impact = 29 to 42 points, high impact = 43 to 56 points, and very high impact = 57 to 70 points. RESULTS: A total of 80 patients were included. The median score obtained in the questionnaire for the sample studied was 34.00 (range=14-68). Statistically significant differences between patients with "primary anal fistula" (n=65) and "recurrent anal fistula" (n=15) were observed (mean rank=42.96 vs. mean rank=29.83, p=0.048). Furthermore, an inverse proportion (P=.016) between "time with clinical symptoms" and "impact on quality of life" was found (5 years: mean rank = 19.00). There were no statistically significant differences (P=.149) between quality of life amongst patients diagnosed with complex (mean rank = 36.13) and simple fistulae (mean rank = 43.59). CONCLUSIONS: Anal fistulae exert moderate-high impact on patients' quality of life. "Shorter time experiencing clinical symptoms" and the "presence of primary fistula" are factors that can be associated with worse quality of life