An in vitro biomolecular breadboard for prototyping synthetic biological circuits

Biomolecular circuit engineering is critical for implementing complex functions in vivo, and is a baseline method in the synthetic biology space. However, current methods for conducting biomolecular circuit engineering are time-consuming and tedious. A complete design-build-test cycle typically takes weeks' to months' time due to the lack of an intermediary between design ex vivo and testing in vivo. In this work, we explore the development and application of a "biomolecular breadboard" composed of an in-vitro transcription-translation (TX-TL) lysate to rapidly speed up the engineering design-build-test cycle. We first developed protocols for creating and using lysates for conducting biological circuit design. By doing so we simplified the existing technology to an affordable ($0.03/uL) and easy to use three-tube reagent system. We then developed tools to accelerate circuit design by allowing for linear DNA use in lieu of plasmid DNA, and by utilizing principles of modular assembly. This allowed the design-build-test cycle to be reduced to under a business day. We then characterized protein degradation dynamics in the breadboard to aid to implementing complex circuits. Finally, we demonstrated that the breadboard could be applied to engineer complex synthetic circuits in vitro and in vivo. Specifically, we utilized our understanding of linear DNA prototyping, modular assembly, and protein degradation dynamics to characterize the repressilator oscillator and to prototype novel three- and five-node negative feedback oscillators both in vitro and in vivo. We therefore believe the biomolecular breadboard has wide application for acting as an intermediary for biological circuit engineering.

Photochemical electron transfer at fixed distance : a synthetic model of the photosynthetic primary process

A series of meso-phenyloctamethylporphyrins covalently bonded at the 4'phenyl position to quinones via rigid bicyclo[2.2.2]octane spacers were synthesized for the study of the dependence of electron transfer reaction rate on solvent, distance, temperature, and energy gap. A general and convergent synthesis was developed based on the condensation of ac-biladienes with masked quinonespacer-benzaldehydes. From picosecond fluorescence spectroscopy emission lifetimes were measured in seven solvents of varying polarity. Rate constants were determined to vary from 5.0x10⁹sec^-1 in N,N-dimethylformamide to 1.15x10¹⁰ Sec^-1 in benzene, and were observed to rise at most by about a factor of three with decreasing solvent polarity. Experiments at low temperature in 2-MTHF glass (77K) revealed fast, nearly temperature-independent electron transfer characterized by non-exponential fluorescence decays, in contrast to monophasic behavior in fluid solution at 298K. This example evidently represents the first photosynthetic model system not based on proteins to display nearly temperature-independent electron transfer at high temperatures (nuclear tunneling). Low temperatures appear to freeze out the rotational motion of the chromophores, and the observed nonexponential fluorescence decays may be explained as a result of electron transfer from an ensemble of rotational conformations. The nonexponentiality demonstrates the sensitivity of the electron transfer rate to the precise magnitude of the electronic matrix element, which supports the expectation that electron transfer is nonadiabatic in this system. The addition of a second bicyclooctane moiety (15 Å vs. 18 Å edge-to-edge between porphyrin and quinone) reduces the transfer rate by at least a factor of 500-1500. Porphyrinquinones with variously substituted quinones allowed an examination of the dependence of the electron transfer rate constant κ_ET on reaction driving force. The classical trend of increasing rate versus increasing exothermicity occurs from 0.7 eV≤ |ΔG^0'(R)| ≤ 1.0 eV until a maximum is reached (κ_ET = 3 x 10⁸ sec^-1 rising to 1.15 x 10¹⁰ sec^-1 in acetonitrile). The rate remains insensitive to ΔG⁰ for ~ 300 mV from 1.0 eV≤ |ΔG^0’(R)| ≤ 1.3 eV, and then slightly decreases in the most exothermic case studied (cyanoquinone, κ_ET = 5 x 10⁹ sec^-1).

HYBRID PHOTOVOLTAIC DEVICES BASED ON ORGANIC SEMICONDUCTING POLYMERS AND INORGANIC NANOSTRUCTURED ELECTRODES

La fotovoltaica orgánica es una tecnología solar emergente que todavía no ha entrado en el mercado. El objetivo de esta tesis ha sido acercar un poco más la industrialización de dicha tecnología mediante el incremento de la eficiencia y la durabilidad de estos dispositivos solares. Para la consecución de dicho objetivo se identificaron las limitaciones existentes y se diseñó una hora de ruta con diversas estrategias para poder superar cada uno de los problemas. Así, mediante un exhaustivo control de la nano morfología del film fotoactivo y la introducción de electrodos nanoestructurados se ha conseguido incrementar la eficiencia. La sustitución de los electrodos estándares por nuevos electrodos basados en óxidos metálicos confiere durabilidad al sistema. Por último, la sustitución del óxido de indio y estaño como electrodo transparente por nanohilos metálicos de plata habilita la posibilidad de fabricar dispositivos solares flexibles de bajo coste.

Initiatives for the improvement of continuous management training

[EN] Purpose. This work aims to present, from the company viewpoint, a structured account of management proposals and practices directed toward improving the intensity and effectiveness of continuous management training (CMT). Design/methodology/approach. The article takes as its main theoretical referents the Theory of Human Capital, the Resource-Based Vision and the contributions made via the new institutional economy with regard to the problems of information asymmetry between companies, employees and training providers and completes the proposals that derive from this theoretical approach. To do this, experience-based contributions are collected from a selection of company training and HR managers from twelve Basque companies characterised by their strong investment in management training. The methodology used was qualitative and obtained by different qualitative techniques: Focus Groups, Nominal Groups and the Delphi Method, which make up the so-called Hybrid Delphi. Findings and implications. The proposals are aimed at the main agents in training activity: training providers, associations and public agents engaged in management training and, particularly, companies themselves. The initiatives seek above all to increase training market transparency, to improve mutual commitments between companies and managers, and to link training and development with culture and strategic management, so that firms make optimal investment in management training. Originality/value. The methodology used is original, and the contributions are consistent with the theory, have a proven practical utility, and are presented in a hierarchy, which facilitates decision making.

A contribuição da Metodologia de Projeto em Design no processo de desenvolvimento de recursos de Tecnologia Assistiva

O presente trabalho apresenta o processo utilizado por terapeutas ocupacionais no desenvolvimento de objetos de Tecnologia Assistiva. São apresentadas definições e teorias relacionadas ao desenvolvimento de produtos de Tecnologia Assistiva a qual trata de um grupo de produtos desenvolvidos para pessoas com deficiência. São apresentados: o design de produtos voltados para inclusão, modelos de ações voltados às pessoas com deficiência, princípios para o desenvolvimento de produtos para pessoas com deficiência, o mercado brasileiro e o perfil do público-alvo, e os métodos e técnicas de desenvolvimento de Tecnologia Assistiva. Sendo o terapeuta ocupacional um profissional que no processo de reabilitação desenvolve produtos de Tecnologia Assistiva, explicita-se o seu papel no desenvolvimento desses produtos. Tendo o Ergodesign como base metodológica para o desenvolvimento de produtos de Tecnologia Assistiva, apresentam-se questões pertinentes à essa teoria dentro da ergonomia e do design. Expõem-se os resultados da pesquisa de campo realizada com terapeutas ocupacionais que produzem produtos de Tecnologia Assistiva, com terapeutas ocupacionais que encaminham para outro profissional realizar a produção, com professores de disciplinas de Tecnologia Assistiva dos cursos de Terapia Ocupacional da cidade do Rio de Janeiro e com usuários desses produtos. A partir da pesquisa bibliográfica e dos resultados obtidos na pesquisa de campo, este trabalho tem como objetivo principal contribuir, através de uma proposta metodológica, para a formação do terapeuta ocupacional no processo de desenvolvimento de objetos para pessoas com deficiência

Auto-ensamblaje de unidades porfirínicas para la obtención de dispositivos supramoleculares

40 p. : il.

Integrated treatment of first episode psychosis with online training (e-learning): study protocol for a randomised controlled trial

Background: The integrated treatment of first episode psychosis has been shown to improve functionality and negative symptoms in previous studies. In this paper, we describe a study of integrated treatment (individual psychoeducation complementary to pharmacotherapy) versus treatment as usual, comparing results at baseline with those at 6-month re-assessment (at the end of the study) for these patients, and online training of professionals to provide this complementary treatment, with the following objectives: 1) to compare the efficacy of individual psychoeducation as add-on treatment versus treatment as usual in improving psychotic and mood symptoms; 2) to compare adherence to medication, functioning, insight, social response, quality of life, and brain-derived neurotrophic factor, between both groups; and 3) to analyse the efficacy of online training of psychotherapists. Methods/design: This is a single-blind randomised clinical trial including patients with first episode psychosis from hospitals across Spain, randomly assigned to either a control group with pharmacotherapy and regular sessions with their psychiatrist (treatment as usual) or an intervention group with integrated care including treatment as usual plus a psychoeducational intervention (14 sessions). Training for professionals involved at each participating centre was provided by the coordinating centre (University Hospital of Alava) through video conferences. Patients are evaluated with an extensive battery of tests assessing clinical and sociodemographic characteristics (Positive and Negative Syndrome Scale, State-Trait Anxiety Inventory, Liebowitz Social Anxiety Scale, Hamilton Rating Scale for Depression, Scale to Assess Unawareness of Mental Disorders, Strauss and Carpenter Prognostic Scale, Global Assessment of Functioning Scale, Morisky Green Adherence Scale, Functioning Assessment Short Test, World Health Organization Quality of Life instrument WHOQOL-BREF (an abbreviated version of the WHOQOL-100), and EuroQoL questionnaire), and brain-derived neurotrophic factor levels are measured in peripheral blood at baseline and at 6 months. The statistical analysis, including bivariate analysis, linear and logistic regression models, will be performed using SPSS. Discussion: This is an innovative study that includes the assessment of an integrated intervention for patients with first episode psychosis provided by professionals who are trained online, potentially making it possible to offer the intervention to more patients.

Charge and spin transport in graphene devices

170 p.

Influence of synthetic method on the properties of La0.5Ba0.5FeO3 SOFC cathode

Power Point presentado en The Energy and Materials Research Conference - EMR2015 celebrado en Madrid (España) entre el 25-27 de febrero de 2015

Mindfulness training for reducing anger, anxiety, and depression in fibromyalgia patients

Fibromyalgia is a disabling syndrome. Results obtained with different therapies are very limited to date.The goal of this study was to verify whether the application of a mindfulnessbased training program was effective in modifying anger, anxiety, and depression levels in a group of women diagnosed with fibromyalgia. This study is an experimental trial that employed a waiting list control group. Measures were taken at three different times: pretest, posttest, and follow-up. The statistical analyses revealed a significant reduction of anger (trait) levels, internal expression of anger, state anxiety, and depression in the experimental group as compared to the control group, as well as a significant increase in internal control of anger. It can be concluded that the mindfulness-based treatment was effective after 7 weeks. These results were maintained 3 months after the end of the intervention.