Gellan gum-hyaluronate spongy-like hydrogels promote angiogenesis in hindlimb ischemia

"Tissue engineering: part A", vol. 21, suppl. 1 (2015)

Exploring silk based biomaterials functionalized with antimicrobial peptides to prevent surgical site infections

Surgical site infections (SSI) often occur after invasive surgery, which is as a serious health problem, making it important to develop new biomaterials to prevent infections. Spider silk is a natural biomaterial with excellent biocompatibility, low immunogenicity and controllable biodegradability. Through recombinant DNA technology, spider silk-based materials can be bioengineered and functionalized with antimicrobial (AM) peptides 1. The aim of this study is to develop new materials by combining spider silk chimeric proteins with AM properties and silk fibroin extracted from Bombyx mori cocoons to prevent microbial infection. Here, spider silk domains derived from the dragline sequence of the spider Nephila clavipes (6 mer and 15 mer) were fused with the AM peptides Hepcidin and Human Neutrophil peptide 1 (HNP1). The spider silk domain maintained its self-assembly features allowing the formation of beta-sheets to lock in structures without any chemical cross-linking. The AM properties of the developed chimeric proteins showed that 6 mer + HNP1 protein had a broad microbicidal activity against pathogens. The 6 mer + HNP-1 protein was then assembled with different percentages of silk fibroin into multifunctional films. In vitro cell studies with a human fibroblasts cell line (MRC5) showed nontoxic and cytocompatible behavior of the films. The positive cellular response, together with structural properties, suggests that this new fusion protein plus silk fibroin may be good candidates as multifunctional materials to prevent SSI.

Production of elastic Chitosan/Chondroitin sulphate multilayer films for tissue regeneration

Membrane-like scaffolds are suitable to induce regeneration in many and different anatomic sites, such as periodontal membrane, skin, liver and cardiac tissues. In some circumstances, the films should adapt to geometrical changes of the attached tissues, such as in cardiac or blood vessel tissue engineering applications. In this context, we developed stretchable two-dimensional multilayer constructs through the assembling of two natural-based polyelectrolytes, chitosan (CHT) and chondroitin sulphate (CS), using the layer-by-layer methodology. The morphology, topography and the transparency of the films were evaluated. The in- fluence of genipin, a natural-derived cross-linker agent, was also investigated in the control of the mechanical properties of the CHT/CS films. The water uptake ability can be tailored by changing the cross-linker concentration, which influenced the young modulus and ultimate tensile strength. The maximum extension tends to decrease with the increase of genipin concentration, compromising the elastic properties of CHT/CS films: nevertheless using lower cross-linker contents, the ultimate tensile stress is similar to the films not cross-linked but exhibiting a significant higher modulus. The in vitro biological assays showed better L929 cell adhesion and proliferation when using the crosslinked membranes and confirmed the non-cytotoxicity of the CHT/CS films. The developed free-standing biomimetic multilayer could be designed to fulfill specific therapeutic requirements by tuning properties such as swelling, mechanical and biological performances.

The use of canonical correlation analysis to assess the relationship between executive functioning and verbal memory in older adults

Executive functioning (EF), which is considered to govern complex cognition, and verbal memory (VM) are constructs assumed to be related. However, it is not known the magnitude of the association between EF and VM, and how sociodemographic and psychological factors may affect this relationship, including in normal aging. In this study, we assessed different EF and VM parameters, via a battery of neurocognitive/psychological tests, and performed a Canonical Correlation Analysis (CCA) to explore the connection between these constructs, in a sample of middle- aged and older healthy individuals without cognitive impairment (N = 563, 50+ years of age). The analysis revealed a positive and moderate association between EF and VM independently of gender, age, education, global cognitive performance level, and mood. These results confirm that EF presents a significant association with VM performance.

A comunicação em organizações virtuais: o paradigma digital

Dissertação de mestrado em Ciências da Comunicação (área de especialização em Publicidade e Relações Públicas)

Desenvolvimento de talentos no desporto : aspectos psicológicos

A presente investigação tem como principal objetivo compreender a relevância de diversos fatores sociodemográficos e psicossociais inerentes ao desenvolvimento do talento em contexto desportivo, numa perspectiva multidimensional. Procedeu-se a uma avaliação quantitativa de jogadores de futebol, integrados num clube de elite, com idades compreendidas entre os 13 e os 19 anos. No sentido de avaliar os construtos psicológicos considerados no presente estudo (motivação, perfecionismo, suporte parental, resiliencia, coping e compromisso) foram utilizados os seguintes questionários: Sport Motivation Scale - SMS (Pelletier et al., 1995); Multidimensional Perfectionism Scale – MPS (Frost, Marten, Lahart, & Rosenblate, 1990); Own Memories of Parental Rearing – EMBU (Perris, Jacobson, Lindstörm, Von Knorring, & Perris, 1980); Resilience Scale – RS (Wagnild & Young, 1993); Athletic Coping Skills – ACSI 28 (Smith, Schutz, Smoll, & Ptacek, 1995); e Elite Athlete Commitment Scale – EACS (Ramadas, Serpa, Rosado, Gouveia & Maroco, 2013). A significância da variável nível de prestação (elite/sub-elite; dispensados/retidos) sobre os diversos constructos psicológicos foi avaliada através da análise da covariância multivariada (MANCOVA), da análise de equações estruturais (CBSEM) e da técnica de míninos quadrados parciais (PLS). Os jogadores mais bem sucedidos (jogadores de elite e jogadores retidos) percecionaram maior suporte parental, demonstraram níveis mais elevados de compromisso, resiliência, autodeterminação, capacidade de adaptação e confronto, assim como um perfeccionismo ajustado. No que concerne às variáveis sociodemográficas, constatou-se que os jogadores retidos jogam predominantemente no segundo ano do respetivo grupo de idade e têm uma idade inferior aos jogadores dispensados. Os resultados obtidos poderão constituir um relevante suporte para futuros programas educacionais que incidam sobre temáticas relacionadas com os compromissos necessários à prossecução e manutenção de níveis de elite, estratégias de coping, gestão da rotina diária, e o papel dos pais no processo de formação do jovem desportista.

Envolvimento parental no desporto

O presente conjunto de investigações pretendeu estudar o envolvimento parental na competição desportiva de crianças e jovens. Baseado no modelo do envolvimento parental no desporto (Teques & Serpa, 2009), o estudo permitiu concretizar dois objectivos fundamentais. Primeiro, desenvolver um conjunto de escalas válidas e fidedignas para aceder aos constructos incluídos no modelo teórico. Segundo, testar as hipóteses fundamentadas na estrutura conceptual do modelo com o propósito de compreender (1) a razão porque os pais se envolvem no desporto dos filhos, (2) quais os comportamentos utilizados pelos pais durante o envolvimento, e (3) como é que o envolvimento influencia o contexto de realização do jovem atleta. No total, participaram voluntariamente 1620 pais e 1665 jovens atletas de vários desportos individuais e coletivos, com idades compreendidas entre os 9 e os 18 anos. A prossecução dos objectivos teve por base uma série de três estudos independentes. Os resultados do primeiro estudo sugerem que as crenças do papel parental, a auto-eficácia, a perceção das invocações oriundas do treinador e do jovem atleta, o tempo e energia disponíveis, e os conhecimentos e competências relacionam-se com as atividades de envolvimento dos pais. No segundo estudo, os resultados demonstraram que as perceções dos comportamentos parentais de encorajamento, reforço, instrução, e modelagem medeiam a relação entre os comportamentos reportados pelos pais e as variáveis psicológicas de auto-eficácia, auto-eficácia social, motivação intrínseca, e estratégias de autorregulação dos jovens. Os resultados do terceiro estudo indicam que as perceções dos comportamentos dos pais relacionam-se com a realização desportiva através dos efeitos de mediação da auto-eficácia, autoeficácia social e das estratégias de autorregulação. Implicações para a intervenção, limitações e direções futuras para a investigação são também discutidas.

A promoção do capital psicológico,work engagement e saúde mental de trabalhadores

Dissertação de Mestrado em Políticas de Desenvolvimento de Recursos Humanos

Construcción de un Test de Conocimiento General. Development of a General Knowledge Test

Entre los factores que contribuyen a predecir el rendimiento académico se pueden destacar aquellos que reflejan capacidades cognitivas (inteligencia, por ejemplo), y aquellas diferencias individuales consideradas como no-cognitivas (rasgos de personalidad, por ejemplo). En los últimos años, también se considera al Conocimiento General (CG) como un criterio para el éxito académico (ver Ackerman, 1997), ya que se ha evidenciado que el conocimiento previo ayuda en la adquisición de nuevo conocimiento (Hambrick & Engle, 2001). Uno de los objetivos de la psicología educacional consiste en identificar las principales variables que explican el rendimiento académico, como también proponer modelos teóricos que expliquen las relaciones existentes entre estas variables. El modelo teórico PPIK (Inteligencia-como-Proceso, Personalidad, Intereses e Inteligencia-como-Conocimiento) propuesto por Ackerman (1996) propone que el conocimiento y las destrezas adquiridas en un dominio en particular son el resultado de la dedicación de recursos cognitivos que una persona realiza durante un prolongado período de tiempo. Este modelo propone que los rasgos de personalidad, intereses individuales/vocacionales y aspectos motivacionales están integrados como rasgos complejos que determinan la dirección y la intensidad de la dedicación de recursos cognitivos sobre el aprendizaje que realiza una persona (Ackerman, 2003). En nuestro medio (Córdoba, Argentina), un grupo de investigadores ha desarrollado una serie de recursos técnicos necesarios para la evaluación de algunos de los constructos propuesto por este modelo. Sin embargo, por el momento no contamos con una medida de Conocimiento General. Por lo tanto, en el presente proyecto se propone la construcción de un instrumento para medir Conocimiento General (CG), indispensable para poder contar con una herramienta que permita establecer parámetros sobre el nivel de conocimiento de la población universitaria y para en próximos trabajos poner a prueba los postulados de la teoría PPIK (Ackerman, 1996). Between the factors that contribute to predict the academic achievement, may be featured those who reflect cognitive capacities (i.g. intelligence) and those who reflect individual differences that are considered like non-cognitive (i.g. personality traits). In the last years, also the General Knowledge has been considered like a criterion for the academic successfully (see Ackerman, 1997), since it has been shown that the previous knowledge helps in the acquisition of the new knowledge (Hambrick & Engle, 2001). An interesting theoretical model that has proposed an explanation for the academic achievement, is the PPIK (intelligence like a process, interests and inteligence like knowledge) proposed by Ackerman (1996), who argues that knowledge and the acquired skills in a particular domain are the result of the dedication of cognitive resources that a person perform during a long period of time. This model proposes that personality traits, individuals interests and motivational aspects are integrated as complex traits that determine the direction and the intensity of the dedication of cognitive resources on the learning that a person make (Ackerman, 2003). In our context, (Córdoba, Argentina), a group of researcher has developed a series of necessary technical resoures for the assesment of some of the theoretical constructs proposed by this model. However, by the moment, we do not have an instrument for evaluate the General Knowledge. Therefore, this project aims the construction of an instrument to asess General Knowledge, essential to set parameters on the knowledge level of the university population and for in next works test the PPIK theory postulates.

Implementation of Reed-Solomon RS (255,239) Code

In this paper the construction of Reed-Solomon RS(255,239) codeword is described and the process of coding and decoding a message is simulated and verified. RS(255,239), or its shortened version RS(224,208) is used as a coding technique in Low-Power Single Carrier (LPSC) physical layer, as described in IEEE 802.11ad standard. The encoder takes 239 8-bit information symbols, adds 16 parity symbols and constructs 255-byte codeword to be transmitted through wireless communication channel. RS(255,239) codeword is defined over Galois Field GF and is used for correcting upto 8 symbol errors. RS(255,239) code construction is fully implemented and Simulink test project is constructed for testing and analyzing purposes.

Human capital in a global and knowledge-based economy. Part 2. Asssessment at the EU country level.

This report is an extension and partial update of de la Fuente and Ciccone (2002). It constructs estimates of the private and social rates of return on schooling for fourteen EU countries using microeconometric estimates of Mincerian wage equations, the results of cross-country growth regressions and OECD data on educational expenditures, tax rates and social benefits. The results are used to draw some tentative conclusions regarding the optimality of observed investment patterns and educational subsidy levels.

L'autoetnografia com a memòria de trànsit

Projecte realitzat en el Departament de dibuix de la Facultat de Belles Arts, a la Universitat de Barcelona, durant el segon curs de Doctorat, en el seminari que a càrrec de Fernando Hernandez on treballem l’ Autoetnografia Narrativa. Aquesta és una investigació que no té un caràcter individual marcat tan sols per la meva veu, sinó que més aviat la configuren també tots els companys de Seminari, amb ells i a través d’ells he aconseguit aprendre molts factors que intento quedin reflectits en el texte que presento i que treballo des de l’Educació Artística i la Cultura Visual. Durant aquest any me n’he adonat de què l’aprenentatge es realitza i té molt a veure amb les vivències que cada un de nosaltres construeix en societat, en elles centro les següents pàgines que per mí són l’inici d’un recorregut cap a un nou coneixement. Intento en tot moment tenir present el grup amb el qual he tingut la possibilitat de treballar durant aquest any i des del qual he pogut treballar-me com a subjecte educadora. D’aquesta manera i des de mí presento la meva contribució a n’aquest construir aprenentatge en societat, a partir de la següent narració.

Coevolution of restriction factors and retroviruses

Les virus exploitent la machinerie cellulaire de l'hôte pour se répliquer. Ils doivent s'adapter pour infecter la cellule hôte de manière optimale tout en échappant à la vigilance du système de défense de l'hôte. Ainsi l'hôte et les virus se livrent à de constantes batailles évolutives. Mon travail de thèse a porté sur l'étude des signatures évolutives de facteurs de l'hôte agissant comme des 'facteurs de restriction' en bloquant la réplication rétrovirale chez les primates. Plus spécifiquement, mon travail a visé à utiliser des données évolutives pour renseigner les analyses fonctionnelles et la biologie. Nous avons étudié le facteur anti-VIH-1 nommé TRIM5a (i) chez les prosimiens pour mieux comprendre son rôle dans le contrôle d'un lentivirus endogène, (ii) dans son activité contre d'autres anciennes infections représentées par des rétrovirus endogènes humains et (iii) en tant que protéine capable de générer des mutants de la capside. Premièrement nous nous sommes intéressés à TRIM5a chez deux espèces de lémuriens dont Microcebus murinus qui porte le lentivirus endogène PSIV dans son génome depuis plusieurs millions d'années,. Nous avons observé que TRIM5a chez M. murinus a un spectre d'activité antivirale réduit à l'opposé de TRIM5a chez le Lemur catta - non porteur du PSIV endogène - qui bloque une large variété de rétrovirus dont le PSIV. De ce fait TRIM5a aurait pu contribuer à protéger certaines espèces de lémuriens vis-à-vis d'anciennes infections par le PSIV. A l'inverse du PSIV, des virus dérivés des rétrovirus endogènes humains HERV-K and HERV-H se sont révélés largement résistants à l'inhibition par TRIM5a. Ces données illustrent une absence de protection par TRIM5a face à d'autres anciennes infections rétrovirales. Puis, pour évaluer l'impact de la protéine TRIM5a humaine sur le VIH-1, nous avons testé l'effet de mutations des résidues sous sélection positive dans la capside du VIH-1 sur l'inhibition par TRIM5a. Nos résultats montrent que TRIM5a ne jouerait pas un rôle significatif dans l'évolution de la capside du VIH-1. Enfin notre travail a porté sur le facteur anti-VIH-1 SAMHD1 récemment découvert, que nous avons séquencé chez 25 espèces de primates. L'analyse évolutive des sites sous sélection positive et des expériences fonctionnelles ont permis d'identifier le domaine de SAMHD1 interagissant avec la protéine lentivirale Vpx. De même que d'autres protéines virales contrecarrent les facteurs de restriction en les menant à la dégradation, nous avons observé que Vpx induit la dégradation de SAMHD1 de manière spécifique à l'espèce. Ces découvertes contribuent à comprendre comment les facteurs de restriction et les virus co-évoluent pour se neutraliser l'un l'autre. - Viruses hijack the host cellular machinery to replicate. They adapt to infect optimally host cells while escaping host defense systems. Viruses and the host coevolve in an evolutionary struggle. My thesis work has been devoted to study the evolutionary signatures of host factors acting as restriction factors that block retroviral replication in primates. Specifically, my work aimed at using evolutionary data to inform functional analyses and biology. We studied the anti-HIV-1 factor TRIM5a (i) in prosimians to better understand its possible role in the control of an endogenous lentivirus, (ii) in its activity against other ancient infections - as represented by HERVs, and (iii) as a protein capable of generating escape mutants in the viral capsid. First, my work focused on two lemur species, one of which was the gray mouse lemur that carries the endogenous lentivirus PSIV integrated in its genome for several million years. TRIM5a from gray mouse lemur exhibited a limited antiviral spectrum as opposed to TRIM5a from ring-tailed lemur - not a host of PSIV - that is able to block diverse retroviruses notably PSIV. These results support the possible contribution of TRIM5a in protecting lemur species from ancient infection by PSIV. In contrast, chimeric viruses derived from two human endogenous retroviruses were broadly resistant to TRIM5a-mediated restriction, suggesting TRIM5a lack of activity against other types of ancient infections. To evaluate the recent impact of human TRIM5a on HIV-1 evolution, we tested whether variants at positively selected sites in the HIV-1 capsid affected the ability of human TRIM5a alleles to restrict HIV-1. Our results indicate that TRIM5a does not play a significant role in the evolution of HIV1 capsid. At last, our work concentrated on the newly discovered anti-HIV-1 restriction factor SAMHD1. We determined its coding sequence in a panel of 25 species of primates. Evolutionary analyses of positively selected sites in SAMHD1 domains and functional assays identified the domain of SAMHD1 interacting with the lentiviral protein Vpx. Similar to other viral countermeasures targeting cellular restriction factors, Vpx was responsible of the degradation of SAMHD1 orthologs in a species-specific manner. These findings contributed to understanding how restriction factors and viruses evolve to counteract each other.

In vitro engineering of human stratified urothelium: analysis of its morphology and function.

PURPOSE: Gastric or intestinal patches, commonly used for reconstructive cystoplasty, may induce severe metabolic complications. The use of bladder tissues reconstructed in vitro could avoid these complications. We compared cellular differentiation and permeability characteristics of human native with in vitro cultured stratified urothelium. MATERIALS AND METHODS: Human stratified urothelium was induced in vitro. Morphology was studied with light and electron microscopy and expression of key cellular proteins was assessed using immunohistochemistry. Permeability coefficients were determined by measuring water, urea, ammonia and proton fluxes across the urothelium. RESULTS: As in native urothelium the stratified urothelial construct consisted of basal membrane and basal, intermediate and superficial cell layers. The apical membrane of superficial cells formed villi and glycocalices, and tight junctions and desmosomes were developed. Immunohistochemistry showed similarities and differences in the expression of cytokeratins, integrin and cellular adhesion proteins. In the cultured urothelium cytokeratin 20 and integrin subunits alpha6 and beta4 were absent, and symplekin was expressed diffusely in all layers. Uroplakins were clearly expressed in the superficial umbrella cells of the urothelial constructs, however, they were also present in intermediate and basal cells. Symplekin and uroplakins were expressed only in the superficial cells of native bladder tissue. The urothelial constructs showed excellent viability, and functionally their permeabilities for water, urea and ammonia were no different from those measured in native human urothelium. Proton permeability was even lower in the constructs compared to that of native urothelium. CONCLUSIONS: Although the in vitro cultured human stratified urothelium did not show complete terminal differentiation of its superficial cells, it retained the same barrier characteristics against the principal urine components. These results indicate that such in vitro cultured urothelium, after being grown on a compliant degradable support or in coculture with smooth muscle cells, is suitable for reconstructive cystoplasty.

A New Large-DNA-Fragment Delivery System Based on Integrase Activity from an Integrative and Conjugative Element.

During the past few decades, numerous plasmid vectors have been developed for cloning, gene expression analysis, and genetic engineering. Cloning procedures typically rely on PCR amplification, DNA fragment restriction digestion, recovery, and ligation, but increasingly, procedures are being developed to assemble large synthetic DNAs. In this study, we developed a new gene delivery system using the integrase activity of an integrative and conjugative element (ICE). The advantage of the integrase-based delivery is that it can stably introduce a large DNA fragment (at least 75 kb) into one or more specific sites (the gene for glycine-accepting tRNA) on a target chromosome. Integrase recombination activity in Escherichia coli is kept low by using a synthetic hybrid promoter, which, however, is unleashed in the final target host, forcing the integration of the construct. Upon integration, the system is again silenced. Two variants with different genetic features were produced, one in the form of a cloning vector in E. coli and the other as a mini-transposable element by which large DNA constructs assembled in E. coli can be tagged with the integrase gene. We confirmed that the system could successfully introduce cosmid and bacterial artificial chromosome (BAC) DNAs from E. coli into the chromosome of Pseudomonas putida in a site-specific manner. The integrase delivery system works in concert with existing vector systems and could thus be a powerful tool for synthetic constructions of new metabolic pathways in a variety of host bacteria.