This dissertation is a composite of some of the research that I have conducted during the course of my PhD study. The larger goal of this dissertation is to renew the interests among the scientific community for an otherwise under-appreciated technique called Isothermal Titration Calorimetry. The resurgence of calorimetry in the biophysical community and the shift to investigations of more complex biological systems signal a real need for more sophisticated analysis techniques. This dissertation expounds on new ITC analysis methods that we have developed as well as results from the study of thermodynamic properties of higher order DNA structures. In 1978, Peter Privalov described the first use of microcalorimetry to obtain the thermodynamic properties for removing calcium from parvalbumin III protein. Fast forward 36 years: modern day electronics, highly efficient thermally conductive and chemically inert materials, in conjunction with sensitive thermal detectors, has transformed the original calorimeter into a device capable of measuring heat changes as small as 0.05 nanowatts, which is equivalent to capturing heat from an incandescent light bulb a kilometer away. However, analytical methods have not kept pace with this technology. Commercial ITC instruments are typically supplied with software that only includes a number of simple interaction models. As a result, the lack of analysis tools for more complex models has become a limiting factor for many researchers. We have recently developed new ITC fitting algorithms that we have incorporated into a userriendly program (CHASM©) for the analysis of complex ITC equilibria. In a little over a year, CHASM© has been downloaded by over 370 unique users. Several chapters in this dissertation demonstrate this software’s power and versatility in the thermodynamic investigations of two model systems in both aqueous and non-aqueous media. In chapter VI, we assembled a model NHE-III1 : a novel structure of Gquadruplex in a double stranded form and studied its structural complexity and binding interactions with a classical G-quaduplex interactive ligand known as TMPyP4. In chapter VII, we reported the thermodynamic properties of a novel PAH system in which weak dispersion forces are solely responsible for formation of the supramolecular complexes.

Lactobacillus kunkeei is one of the most abundant bacteria within the honey crop of the honey bee. Genome sequencing of L. kunkeei isolated from honey bees all over the world showed several genes unique for L. kunkeei. Among these orphan genes, an array of four to five highly conserved genes coding for giant extracellular proteins were found. Cryogenic electron microscopy imaging of a giant-protein preparation from L. kunkeei A00901 showed an overall structure similar to a long string with a knot at the end. Further analysis showed high similarity between the different giants at the N-terminus, and secondary structure predictions showed that the same region was rich in β-sheets.  These results, combined with the knowledge of other large extracellular proteins, led to the hypothesis that the “knot” domain is located at the N-terminus and that these proteins are used by the cell to latch on to the intestine lining or other cells in the honey crop. In this study, predictions were made to locate the N-terminal domains of two of these giant proteins. Four different constructs were made for each protein, where three constructs were designed for expression and purification of the N-terminal domain with different end-positions, and one construct was for a predicted β-solenoid domain located downstream from the N-terminal domain. The protein constructs were recombinantly produced in E. coli, and three of the N-terminal constructs from both proteins were purified. Thermal stability was tested using nano differential scanning fluorimetry (nanoDSF), Thermofluor, and circular dichroism (CD), which all showed characteristic melting curves at low melting temperatures, ranging from 33 °C to 44 °C, for all three constructs. During CD measurements, all three constructs showed refolding after thermal denaturation and a higher abundance of antiparallel β-sheets over α-helices. Looking at the protein structure, small angle X-ray scattering data indicated that all three proteins formed elongated structures. These results indicate that a folded domain has been found for both proteins. Although, further analysis will be required to determine the boundaries of the N-terminal domains, and to elucidate if these domains have anything to do with ligand binding and the L. kunkeei ability to latch onto the honey crop.

No description available.

No description available.

This thesis aims to investigate whether additive manufacturing is applicable in manufacturing the 1N thruster option that Bradford Ecaps offers. Therefore, the nozzle design is of particular interest as AM provides accessibilities to manufacturing complex structures. The current Ecaps 1N thruster has an operating thrust lifespan that exceeds the required lifespan commonly needed for the majority of customers. With AM, an increase in production throughput and optimization of nozzle design is possible. A candidate material, a platinum group metal, was picked for a future 1N thruster prototype concerning the limiting operating constraints. Computational fluid analysis was performed to investigate different contour nozzles to investigate the possibility of improving the performance of the Bradford ECAPS 1N thruster. AMATLAB code was developed to model the contour nozzles, and ANSYS Fluent was used for the computational analysis. Three different nozzle geometries were evaluated to investigate the overall performance of the expanding exhaust gas and thrust properties in vacuum conditions. Configuration 1. which had an extended nozzle was selected as a solution since it eliminatedthe interferences with the continuum. The Nasa CEA code was used to generate the fluid gas properties. No substantial performance gain was observed for the 1N thruster. This was found to be due to the boundary-dominated flow exhibited in the nozzle. A conical nozzle was found to work comparatively well. / Detta examensarbete syftar till att undersöka om additiv tillverkning (AM) är tillämplig vid tillverkning av Bradford Ecaps 1N raketmotor. Därför är munstycksdesignen av särskilt intresse eftersom AM ger möjlighet för tillverkning av komplexa strukturer. Den nuvarande Ecaps 1N-motorn har en livslängd som överstiger den livslängd som krävs för de flesta kunder. Med AM är ökning av produktionsgenomströmningen och optimering av munstycksdesign möjlig. Ett kandidatmaterial, en metall i platinagruppen, valdes ut för en framtida 1N prototyp med hänvisning till de begränsande driftsbegränsningarna. Beräkningsflödesanalys utfördes för att undersöka olika konturmunstycken för att undersöka möjligheten till att förbättra prestandan hos Bradford ECAPS 1N framdrivningssystem. En MATLAB-kod utvecklades för att modellera konturmunstyckena och ANSYS Fluent användes för beräkningsanalysen. Tre olika munstycksgeometrier utvärderades för att undersöka den totala prestandan hos de expanderande avgaserna och dragkraftsegenskaperna under vakuumförhållanden. Konfiguration 1. som hade ett förlängt munstycke valdes som en lösning eftersom detta eliminerade interferenserna med kontinuumet. Nasa CEA-koden användes för att generera fluidens gasegenskaper. Ingen betydande prestandaökning observerades för 1N motorn. Detta visade sig bero på det gränsskiktsdominerade flödet som uppvisades i munstycket. Ett koniskt munstycke visade sig fungera relativt bra.

For the last ten years, DVD-R media have played an important role in the storage of large amounts of digital data throughout the world. During this time it was assumed that the DVD-R was as long-lasting and stable as its predecessor, the CD-R. Several reports have surfaced over the last few years questioning the DVD-R's ability to maintain many of its claims regarding archival quality life spans. These reports have shown a wide range of longevity between the different brands. While some DVD-Rs may last a while, others may result in an early and unexpected failure. Compounding this problem is the lack of information available for consumers to know the quality of the media they own. While the industry works on devising a standard for labeling the quality of future media, it is currently up to the consumer to pay close attention to their own DVD-R archives and work diligently to prevent data loss. This research shows that through accelerated aging and the use of logistic regression analysis on data collected through periodic monitoring of disc read-back errors it is possible to accurately predict unrecoverable failures in the test discs. This study analyzed various measurements of PIE errors, PIE8 Sum errors, POF errors and jitter data from three areas of the disc: the whole disc, the region of the disc where it first failed as well as the last half of the disc. From this data five unique predictive equations were produced, each with the ability to predict disc failure. In conclusion, the relative value of these equations for end-of-life predictions is discussed.

Flax (Linum usitatissimum L.) is a self-pollinated crop widely cultivated for fiber and oil production. Flaxseed is renowned for its health attributes but the presence of compounds, such as the heavy metal cadmium (Cd), is undesirable. Genomic studies in flax have produced large amounts of data in the last 15 years, providing useful resources to improve the genetic of this crop using genomics-based technologies and strategies. The goal of this thesis is therefore to capitalize on these advances to address the Cd problem and to propose solutions to improve breeding efficiencies. To find genomic-based solutions to Cd content, to the currently low breeding efficiency and to abiotic stress resistance in flax, this study utilized four major strategies: (1) genomic cross prediction, (2) gene family identification, (3) genome-wide association study (GWAS) and (4) genomic selection (GS). Characterization of the ATP-binding cassette (ABC) transporter and heavy metal associated (HMA) gene families was performed using the flax genome sequence. A total of 198 ABC transporter and 12 HMA genes were identified in the flax genome, of which nine were orthologous to Cd-associated genes in Arabidopsis, rice and maize. A transcriptomic analysis of eight tissues provided some support towards the functional annotation of these genes and confirmed the expression of these ABC transporter and HMA genes in flax seeds and other tissues. A diversity panel of 168 flax accessions was grown in the field at multiple locations and years and the seed content of 24 heavy metals (HMs) was measured. The panel was also sequenced and a single nucleotide polymorphism (SNP) dataset of nearly 43,000 SNPs was defined. A GWAS was conducted using these genotypic and phenotypic data and a total of 355 non-redundant quantitative trait nucleotides (QTNs) were identified for ten of the 24 metal contents. Overall, a total of 24 major and 331 minor effect QTNs were detected, including 11 that were pleiotropic. After allelic tests, 108 non-redundant QTNs were retained for eight of the ten metals and ranging from one for copper (Cu) to 70 for strontium (Sr). A total of 20 candidate genes for HM accumulation were identified at 12 of the 24 major QTN loci, of which five belonged to the ABC transporter family. Many of the metal contents, including Cd, appeared to be controlled by many genes of small effects; hence, GS is better suited than marker-assisted selection for application in breeding. To test this, predictive ability using ten GS statistical models was evaluated using trait-specific QTN and the random genome-wide 43K SNP datasets. Significantly higher predictive abilities were observed from the GS models built with the dataset made of QTNs associated with metal contents (70-80%) compared to that of the 43K dataset (10-25%). This study showed the feasibility of using GS to improve the predictive ability of polygenic traits such as metal content in seeds. GS can be applied in early generation selection to accelerate the improvement of abiotic stress resistance and either select low-Cd lines or discard high-Cd lines. These findings validate the use of a QTL-based strategy as a highly effective method for improving the efficiency of predictive ability of GS for highly complex traits such as resistance or tolerance to HM accumulation. Identification of both large and minor effect QTNs and/or pleiotropic effects hold potential for flax breeding improvement. Candidate gene functional validation can be performed using methods such as genome editing or targeting induced local lesions in genomes (TILLING).

No description available.

No description available.

No description available.