966 resultados para SINGLE-CELL ASSAY
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Massive multi-user multiple-input multiple-output (MU-MIMO) systems are cellular networks where the base stations (BSs) are equipped with hundreds of antennas, N, and communicate with tens of mobile stations (MSs), K, such that, N ≫ K ≫ 1. Contrary to most prior works, in this paper, we consider the uplink of a single-cell massive MIMO system operating in sparse channels with limited scattering. This case is of particular importance in most propagation scenarios, where the prevalent Rayleigh fading assumption becomes idealistic. We derive analytical approximations for the achievable rates of maximum-ratio combining (MRC) and zero-forcing (ZF) receivers. Furthermore, we study the asymptotic behavior of the achievable rates for both MRC and ZF receivers, when N and K go to infinity under the condition that N/K → c ≥ 1. Our results indicate that the achievable rate of MRC receivers reaches an asymptotic saturation limit, whereas the achievable rate of ZF receivers grows logarithmically with the number of MSs.
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Acid stimulated accumulation of insoluble phosphorus within microbial cells is highly beneficial to wastewater treatment but remains largely unexplored. Using single cell analyses and next generation sequencing, the response of active polyphosphate accumulating microbial communities under conditions of enhanced phosphorus uptake under both acidic and aerobic conditions was characterised. Phosphorus accumulation activities were highest under acidic conditions (pH 5.5 > 8.5), where a significant positive effect on bioaccumulation was observed at pH 5.5 when compared to pH 8.5. In contrast to the Betaproteobacteria and Actinobacteria dominated enhanced biological phosphorus removal process, the functionally active polyP accumulators at pH 5.5 belonged to the Gammaproteobacteria, with key accumulators identified as members of the families Aeromonadaceae and Enterobacteriaceae. This study demonstrated a significant enrichment of key polyphosphate kinase and exopolyphosphatase genes within the community metagenome after acidification, concomitant with an increase in P accumulation kinetics.
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Thesis (Ph.D.)--University of Washington, 2016-08
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Single-cell oils (SCO) have been considered a promising source of 3rd generation biofuels mainly in the final form of biodiesel. However, its high production costs have been a barrier towards the commercialization of this commodity. The fast growing yeast Rhodosporidium toruloides NCYC 921 has been widely reported as a potential SCO producing yeast. In addition to its well-known high lipid content (that can be converted into biodiesel), is rich in high value added products such as carotenoids with commercial interest. The process design and integration may contribute to reduce the overall cost of biofuels and carotenoid production and is a mandatory step towards their commercialization. The present work addresses the biomass disruption, extraction, fractionation and recovery of products with special emphasis on high added valued carotenoids (beta-carotene, torulene, torularhodin) and fatty acids directed to biodiesel. The chemical structure of torularhodin with a terminal carboxylic group imposes an additional extra challenge in what concern its separation from fatty acids. The proposed feedstock is fresh biomass pellet obtained directly by centrifugation from a 5L fed-batch fermentation culture broth. The use of a wet instead of lyophilised biomass feedstock is a way to decrease processing energy costs and reduce downstream processing time. These results will contribute for a detailed process design. Gathered data will be of crucial importance for a further study on Life-Cycle Assessment (LCA).
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Nesta tese procurou-se demonstrar a valoração do efluente do processamento de pescado por incorporação dos nutrientes em Aphanothece microscopica Nägeli a diferentes temperaturas. Para tanto o trabalho é composto de cinco artigos que objetivaram avaliar sob o ponto de vista do tratamento do efluente pela cianobactéria Aphanothece e a separação e avaliação da biomassa gerada. O primeiro artigo intitula-se “Influência da temperatura na remoção de nutrientes do efluente da indústria de pescado por Aphanothece microscopica Nägeli”, e teve por objetivo avaliar a influência da temperatura (10, 20 e 30ºC) em um sistema de tratamento pela cianobactéria Aphanothece na remoção de matéria orgânica, nitrogênio e fósforo do efluente oriundo do processamento de pescado. A análise dos resultados mostrou que a temperatura influenciou significativamente na remoção de DQO, NTK, N-NH4 + e P-PO4 -3 . Para os experimentos a 20 e 30ºC todos os limites estabelecidos para os parâmetros avaliados foram atingidos. O segundo artigo intitulado “Efeito de coagulantes no efluente da indústria da pesca visando à separação de biomassa quando tratado por cianobactéria” avaliou o efeito da concentração e pH de dois tipos de coagulantes, cloreto férrico (FeCl3) e sulfato de alumínio (Al2(SO4)3), na separação da biomassa da cianobactéria Aphanothece microscopica Nägeli cultivada em efluente da indústria da pesca, assim como a remoção de matéria orgânica e nutrientes do efluente. Os resultados indicaram que o coagulante FeCl3 foi mais eficaz na remoção de todos os parâmetros testados. No que concerne à separação da biomassa, com um número de seis lavagens foi removido cerca de 97,6% da concentração de FeCl3 adicionado inicialmente. O terceiro artigo com o título “Caracterização da biomassa de Aphanothece microscopica Nägeli gerada no efluente da indústria da pesca em diferentes temperaturas de cultivo” avaliou a composição química da biomassa da cianobactéria Aphanothece microscopica Nägeli quando desenvolvida em meio de cultivo padrão BG11 e no efluente do processamento de pescado. O quarto artigo teve como título “Influência do meio de cultivo e temperatura em compostos nitrogenados na cianobactéria Aphanothece microscopica Nägeli” objetivou avaliar o teor de compostos nitrogenados presentes na biomassa da cianobactéria Aphanothece microscopica Nägeli quando cultivada em meio padrão e no efluente da indústria da pesca nas diferentes fases de crescimento. Para o estudo da composição química e nitrogenados no efluente foram realizados experimentos nas temperaturas de 10, 20 e 30ºC. As concentrações de proteína, cinzas e pigmentos aumentaram com o aumento da temperatura. Por outro lado, foi observada uma redução do teor de lipídios e carboidratos com o aumento da temperatura. O íon amônio juntamente com os ácidos nucléicos representa uma importante fração do nitrogênio não protéico presente na biomassa da cianobactéria Aphanothece. Ficou demonstrada a influência do meio de cultivo na concentração de nitrogênio, bem como a determinação de proteína pelo método de Kjeldahl superestima a concentração protéica em cianobactérias. O quinto artigo intitulado “Produção de proteína unicelular a partir do efluente do processamento do pescado: modelagem preditiva e simulação” avaliou a produção de proteína unicelular através do cultivo da cianobactéria Aphanothece microscopica Nägeli no efluente da indústria da pesca. Os dados cinéticos de crescimento celular foram ajustados a quatro modelos matemáticos (Logístico, Gompertz, Gompertz Modificado e Baranyi). Os resultados demonstraram que o modelo Logístico foi considerado o mais adequado para descrever a formação de biomassa. A análise preditiva mostrou a possibilidade da obtenção de 1,66, 18,96 e 57,36 kg.m-3.d-1 de biomassa por volume do reator em 1000 h de processo contínuo, para as temperaturas de 10, 20 e 30ºC, respectivamente.
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Autologous nerve grafts are the current gold standard for the repair of peripheral nerve injuries. However, there is a need to develop an alternative to this technique, as donor-site morbidities such as neuroma formation and permanent loss of function are a few of the limitations concerned with this technique. Artificial nerve conduits have therefore emerged as an alternative for the repair of short peripheral nerve defects of less than 30 mm, however they do not surpass autologous nerve grafts clinically. To develop a nerve conduit that supports regeneration over long nerve gaps and in large diameter nerves, researchers have focused on functionalizing of the conduits by studying the components that enhance nerve regeneration such as micro/nano-topography, growth factor delivery systems, supportive cells and extracellular matrix (ECM) proteins as well as understanding the complex biological reactions that take place during peripheral nerve regeneration. This thesis presents strategies to improve peripheral nerve interfaces to better the regenerative potential by using dorsal root ganglions (DRGs) isolated from neonatal rats as an in vitro model of nerve regeneration. The work started off by investigating the usefulness of a frog foam protein Ranaspumin-2 (Rsn2) to coat biomaterials for compatibility, this lead to the discovery of temporary cell adhesion on polydimethylsiloxane (PDMS), which was investigated as a suitable tool to derive cell-sheets for nerve repair. The influence of Rsn2 anchored to specific adhesion peptide sequences, such as isoleucine-lysine-valine-alanine-valine (IKVAV), a sequence derived from laminin proven to promote cell adhesion and neurite outgrowth, was tested as a useful means to influence nerve regeneration. This approach improves the axonal outgrowth and maintains outgrowth long term. Based on the hypothesis that combinational modulation of substrate topography, stiffness and neurotrophic support, affects axonal outgrowth in whole DRGs, dissociated DRGs were used to assess if these factors similarly act at the single cell level. Rho associated protein kinase (ROCK) and myosin II inhibitors, which affect cytoskeletal contractility, were used to influence growth cone traction forces and have shown that these factors work in combination by interfering with growth cone dynamic creating a different response in axonal outgrowth at the single cell level.
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The marine dinoflagellate genus Dinophysis includes species that are the causative agents of diarrhetic shellfish poisoning (DSP). Recent findings indicate that some Dinophysis species are mixotrophic, i.e. capable of both autotrophic and heterotrophic nutrition. We investigated inorganic (and organic) carbon uptake by several species of Dinophysis in the Light and dark using the 'single-cell C-14 method', and compared uptake rates with those of photosynthetic Ceratium species and heterotrophic dinoflagellates in the genus Protoperidinium. Experiments were conducted with water from the Gullmar Fjord and from the Koster Strait (Swedish west coast). Nutrient-enriched phytoplankton from surface water samples were concentrated (20 to 70 mu m) and incubated at in situ temperature under artificial light conditions with high concentrations of inorganic C-14 (1 mu Ci ml(-1)). Individual cells of each desired species were manually isolated under a microscope and transferred to scintillation vials. C. tripes showed net C-14 uptake only during light periods, whereas both C. lineatum and C. furca showed C-14 uptake in the Light as well as uptake (and sometimes losses) in the dark. Dinophysis species had similar carbon fixation rates in Light compared to Ceratium species. For D. acuminata and D. norvegica, net carbon uptake occurred in both Light and dark periods. D. acuta showed a loss of carbon in the dark in one experiment, but in another, dark C uptake was significantly higher than uptake in Light. When exposed to Light, C. furca, D. norvegica and D. acuta had high specific carbon uptake rates. Growth rates for the different species were calculated from C-14 uptake by the cells during the first hours of incubation in light. D. acuminata and D. norvegica had similar maximum growth rates, 0.59 and 0.63 d(-1) (mu); the maximum growth rate of D. acuta was lower (0.41 d(-1)). The positive dark carbon uptake by Dinophysis may suggest a mixotrophic mode of nutrition. In one experiment, both D. norvegica and D. acuta showed a significantly higher carbon uptake in a dark bottle than in a Light bottle, which would be consistent with uptake of C-14-labeled organic matter by D. norvegica and D. acuta. Demonstration of direct uptake of dissolved and particulate organic matter would provide conclusive evidence of mixotrophy and this will require the development of new protocols for measuring organic matter uptake applicable to Dinophysis in the natural assemblages.
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Dissertação (mestrado)—Universidade de Brasília, Instituto de Química, Programa de Pós-Graduação em Química, 2015.
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Dissertação (mestrado)—Universidade de Brasília, Instituto de Química, Programa de Pós-Graduação em Química, 2015.
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International audience
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Biodiesel is a fuel obtained from vegetable oils, such as soy, castorbean, among others. The monoester of fatty acid of these oils have chains with mono, di and tri double connections. The presence of these insaturations are susceptible to oxidization. Antioxidants are substances able to prevent oxidization from oils, fats, fat foods, as well as esters of Alquila( biodiesel). The objective of this work is to summarize a new antioxidant from the Cashew Nut Shell Liquid (CNSL) using the electrolysis technique. A current of 2 amperes was used in a single cell of only one group and two eletrodos of stainless steel 304 in a solution of methanol, together with the eletrolits: acetic acid, sodium chloride and sodium hydroxide, for two hours of agitation. The electrolysis products are characterized by the techniques of cromatography in a thin layer, spectroscopy of infrared and gravimetric analysis. The material was submitted to tests of oxidative stability made by the techniques of spectropy of impendancy and Rancimat (EN 14112). The analyses of characterization suggest that the polimerization of the electrolytic material ocurred. The application results of these materials as antioxidants of soy biodiesel showed that the order of the oxidative stability was obtained by both techniques used
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Alcohol is one of the oldest and most widely used drugs on the planet, but the cellular mechanisms by which it affects neural function are still poorly understood. Unlike other drugs of abuse, alcohol has no specific receptor in the nervous system, but is believed to operate through GABAergic and serotonergic neurotransmitter systems. Invertebrate models offer circuits of reduced numerical complexity and involve the same cell types and neurotransmitter systems as vertebrate circuits. The well-understood neural circuits controlling crayfish escape behavior offer neurons that are modulated by GABAergic inhibition, thus making tail-flip circuitry an effective circuit model to study the cellular mechanisms of acute alcohol exposure. Crayfish are capable of two stereotyped, reflexive escape behaviors known as tail-flips that are controlled by two different pairs of giant interneurons, the lateral giants (LG) and the medial giants (MG). The LG circuit has been an established model in the neuroscience field for more than 60 years and is almost completely mapped out. In contrast, the MG is still poorly understood, but has important behavioral implications in social behavior and value-based decision making. In this dissertation, I show that both crayfish tail-flip circuitry are physiologically sensitive to relevant alcohol concentrations and that this sensitivity is observable on the single cell level. I also show that this ethyl alcohol (EtOH) sensitivity in the LG can be changed by altering the crayfish’s recent social experience and by removing descending inputs to the LG. While the MG exhibits similar physiological sensitivity, its inhibitory properties have never been studied before this research. Through the use of electrophysiological and pharmacological techniques, I show that the MG exhibits many similar inhibitory properties as the LG that appear to be the result of GABA-mediated chloride currents. Finally, I present evidence that the EtOH-induced changes in the MG are blocked through pre-treatment of the potent GABAA receptor agonist, muscimol, which underlines the role of GABA in EtOH’s effects on crayfish tail-flip circuitry. The work presented here opens the way for crayfish tail-flip circuitry to be used as an effective model for EtOH’s acute effects on aggression and value-based decision making.
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Synthetic biology, by co-opting molecular machinery from existing organisms, can be used as a tool for building new genetic systems from scratch, for understanding natural networks through perturbation, or for hybrid circuits that piggy-back on existing cellular infrastructure. Although the toolbox for genetic circuits has greatly expanded in recent years, it is still difficult to separate the circuit function from its specific molecular implementation. In this thesis, we discuss the function-driven design of two synthetic circuit modules, and use mathematical models to understand the fundamental limits of circuit topology versus operating regimes as determined by the specific molecular implementation. First, we describe a protein concentration tracker circuit that sets the concentration of an output protein relative to the concentration of a reference protein. The functionality of this circuit relies on a single negative feedback loop that is implemented via small programmable protein scaffold domains. We build a mass-action model to understand the relevant timescales of the tracking behavior and how the input/output ratios and circuit gain might be tuned with circuit components. Second, we design an event detector circuit with permanent genetic memory that can record order and timing between two chemical events. This circuit was implemented using bacteriophage integrases that recombine specific segments of DNA in response to chemical inputs. We simulate expected population-level outcomes using a stochastic Markov-chain model, and investigate how inferences on past events can be made from differences between single-cell and population-level responses. Additionally, we present some preliminary investigations on spatial patterning using the event detector circuit as well as the design of stationary phase promoters for growth-phase dependent activation. These results advance our understanding of synthetic gene circuits, and contribute towards the use of circuit modules as building blocks for larger and more complex synthetic networks.
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Efficient numerical models facilitate the study and design of solid oxide fuel cells (SOFCs), stacks, and systems. Whilst the accuracy and reliability of the computed results are usually sought by researchers, the corresponding modelling complexities could result in practical difficulties regarding the implementation flexibility and computational costs. The main objective of this article is to adapt a simple but viable numerical tool for evaluation of our experimental rig. Accordingly, a model for a multi-layer SOFC surrounded by a constant temperature furnace is presented, trained and validated against experimental data. The model consists of a four-layer structure including stand, two interconnects, and PEN (Positive electrode-Electrolyte-Negative electrode); each being approximated by a lumped parameter model. The heating process through the surrounding chamber is also considered. We used a set of V-I characteristics data for parameter adjustment followed by model verification against two independent sets of data. The model results show a good agreement with practical data, offering a significant improvement compared to reduced models in which the impact of external heat loss is neglected. Furthermore, thermal analysis for adiabatic and non-adiabatic process is carried out to capture the thermal behaviour of a single cell followed by a polarisation loss assessment. Finally, model-based design of experiment is demonstrated for a case study.