989 resultados para Digestive enzymes Testing


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Smart and mobile environments require seamless connections. However, due to the frequent process of ''discovery'' and disconnection of mobile devices while data interchange is happening, wireless connections are often interrupted. To minimize this drawback, a protocol that enables an easy and fast synchronization is crucial. Bearing this in mind, Bluetooth technology appears to be a suitable solution to carry on such connections due to the discovery and pairing capabilities it provides. Nonetheless, the time and energy spent when several devices are being discovered and used at the same time still needs to be managed properly. It is essential that this process of discovery takes as little time and energy as possible. In addition to this, it is believed that the performance of the communications is not constant when the transmission speeds and throughput increase, but this has not been proved formally. Therefore, the purpose of this project is twofold: Firstly, to design and build a framework-system capable of performing controlled Bluetooth device discovery, pairing and communications. Secondly, to analyze and test the scalability and performance of the \emph{classic} Bluetooth standard under different scenarios and with various sensors and devices using the framework developed. To achieve the first goal, a generic Bluetooth platform will be used to control the test conditions and to form a ubiquitous wireless system connected to an Android Smartphone. For the latter goal, various stress-tests will be carried on to measure the consumption rate of battery life as well as the quality of the communications between the devices involved.

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Synthetic biology promises to transform organic synthesis by enabling artificial catalysis in living cells. I start by reviewing the state of the art in this young field and recognizing that new approaches are required for designing enzymes that catalyze nonnatural reactions, in order to expand the scope of biocatalytic transformations. Carbene and nitrene transfers to C=C and C-H bonds are reactions of tremendous synthetic utility that lack biological counterparts. I show that various heme proteins, including cytochrome P450BM3, will catalyze promiscuous levels of olefin cyclopropanation when provided with the appropriate synthetic reagents (e.g., diazoesters and styrene). Only a few amino acid substitutions are required to install synthetically useful levels of stereoselective cyclopropanation activity in P450BM3. Understanding that the ferrous-heme is the active species for catalysis and that the artificial reagents are unable to induce a spin-shift-dependent increase in the redox potential of the ferric P450, I design a high-potential serine-heme ligated P450 (P411) that can efficiently catalyze cyclopropanation using NAD(P)H. Intact E. coli whole-cells expressing P411 are highly efficient asymmetric catalysts for olefin cyclopropanation. I also show that engineered P450s can catalyze intramolecular amination of benzylic C-H bonds from arylsulfonyl azides. Finally, I review other examples of where synthetic reagents have been used to drive the evolution of novel enzymatic activity in the environment and in the laboratory. I invoke preadaptation to explain these observations and propose that other man-invented reactions may also be transferrable to natural enzymes by using a mechanism-based approach for choosing the enzymes and the reagents. Overall, this work shows that existing enzymes can be readily adapted for catalysis of synthetically important reactions not previously observed in nature.

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Experimental research on a 150 kW arc-heated plasma testing facility was conducted. Stable plasma jets with different gas compositions, temperatures and velocities were obtained at chamber pressure between 400 Pa – 100 kPa. Stagnation ablation experiments were conducted on samples of typical super alloys used for thermal protection systems. The microstructure and hardness of alloys before and after ablation were compared.

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Recent observations of the temperature anisotropies of the cosmic microwave background (CMB) favor an inflationary paradigm in which the scale factor of the universe inflated by many orders of magnitude at some very early time. Such a scenario would produce the observed large-scale isotropy and homogeneity of the universe, as well as the scale-invariant perturbations responsible for the observed (10 parts per million) anisotropies in the CMB. An inflationary epoch is also theorized to produce a background of gravitational waves (or tensor perturbations), the effects of which can be observed in the polarization of the CMB. The E-mode (or parity even) polarization of the CMB, which is produced by scalar perturbations, has now been measured with high significance. Con- trastingly, today the B-mode (or parity odd) polarization, which is sourced by tensor perturbations, has yet to be observed. A detection of the B-mode polarization of the CMB would provide strong evidence for an inflationary epoch early in the universe’s history.

In this work, we explore experimental techniques and analysis methods used to probe the B- mode polarization of the CMB. These experimental techniques have been used to build the Bicep2 telescope, which was deployed to the South Pole in 2009. After three years of observations, Bicep2 has acquired one of the deepest observations of the degree-scale polarization of the CMB to date. Similarly, this work describes analysis methods developed for the Bicep1 three-year data analysis, which includes the full data set acquired by Bicep1. This analysis has produced the tightest constraint on the B-mode polarization of the CMB to date, corresponding to a tensor-to-scalar ratio estimate of r = 0.04±0.32, or a Bayesian 95% credible interval of r < 0.70. These analysis methods, in addition to producing this new constraint, are directly applicable to future analyses of Bicep2 data. Taken together, the experimental techniques and analysis methods described herein promise to open a new observational window into the inflationary epoch and the initial conditions of our universe.

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Motivated by recent MSL results where the ablation rate of the PICA heatshield was over-predicted, and staying true to the objectives outlined in the NASA Space Technology Roadmaps and Priorities report, this work focuses on advancing EDL technologies for future space missions.

Due to the difficulties in performing flight tests in the hypervelocity regime, a new ground testing facility called the vertical expansion tunnel is proposed. The adverse effects from secondary diaphragm rupture in an expansion tunnel may be reduced or eliminated by orienting the tunnel vertically, matching the test gas pressure and the accelerator gas pressure, and initially separating the test gas from the accelerator gas by density stratification. If some sacrifice of the reservoir conditions can be made, the VET can be utilized in hypervelocity ground testing, without the problems associated with secondary diaphragm rupture.

The performance of different constraints for the Rate-Controlled Constrained-Equilibrium (RCCE) method is investigated in the context of modeling reacting flows characteristic to ground testing facilities, and re-entry conditions. The effectiveness of different constraints are isolated, and new constraints previously unmentioned in the literature are introduced. Three main benefits from the RCCE method were determined: 1) the reduction in number of equations that need to be solved to model a reacting flow; 2) the reduction in stiffness of the system of equations needed to be solved; and 3) the ability to tabulate chemical properties as a function of a constraint once, prior to running a simulation, along with the ability to use the same table for multiple simulations.

Finally, published physical properties of PICA are compiled, and the composition of the pyrolysis gases that form at high temperatures internal to a heatshield is investigated. A necessary link between the composition of the solid resin, and the composition of the pyrolysis gases created is provided. This link, combined with a detailed investigation into a reacting pyrolysis gas mixture, allows a much needed consistent, and thorough description of many of the physical phenomena occurring in a PICA heatshield, and their implications, to be presented.

Through the use of computational fluid mechanics and computational chemistry methods, significant contributions have been made to advancing ground testing facilities, computational methods for reacting flows, and ablation modeling.

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The creation of novel enzyme activity is a great challenge to protein engineers, but nature has done so repeatedly throughout the process of natural selection. I begin by outlining the multitude of distinct reactions catalyzed by a single enzyme class, cytochrome P450 monooxygenases. I discuss the ability of cytochrome P450 to generate reactive intermediates capable of diverse reactivity, suggesting this enzyme can also be used to generate novel reactive intermediates in the form of metal-carbenoid and nitrenoid species. I then show that cytochrome P450 from Bacillus megaterium (P450BM3) and its isolated cofactor can catalyze metal-nitrenoid transfer in the form of intramolecular C–H bond amination. Mutations to the protein sequence can enhance the reactivity and selectivity of this transformation significantly beyond that of the free cofactor. Next, I demonstrate an intermolecular nitrene transfer reaction catalyzed by P450BM3 in the form of sulfide imidation. Understanding that sulfur heteroatoms are strong nucleophiles, I show that increasing the sulfide nucleophilicity through substituents on the aryl sulfide ring can dramatically increase reaction productivity. To explore engineering nitrenoid transfer in P450BM3, active site mutagenesis is employed to tune the regioselectivity intramolecular C–H amination catalysts. The solution of the crystal structure of a highly selective variant demonstrates that hydrophobic residues in the active site strongly modulate reactivity and regioselectivity. Finally, I use a similar strategy to develop P450-based catalysts for intermolecular olefin aziridination, demonstrating that active site mutagenesis can greatly enhance this nitrene transfer reaction. The resulting variant can catalyze intermolecular aziridination with more than 1000 total turnovers and enantioselectivity of up to 99% ee.

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The material included within this report is the result of a series of tests of concrete specimens taken during the construction of various buildings in the cities of Pasadena and Los Angeles over a period of eight months.

The object of the problem is to determine the effect of the water ratio on the ultimate strength of the concrete as obtained from data observed and recorded from specimens taken from actual building practice rather than that from laboratory specimens made under ideal, or at least more nearly standard conditions.