EXPLORATIONS IN HOMEOVISCOUS ADAPTATION AND MASS SPECTRAL ANALYSIS OF MEMBRANE LIPIDS

The focus of this dissertation is centered on the mass spectral analysis of lipids and changes occurring in keeping with the concept of homeoviscous adaptation [1]. Homeoviscous adaptation is the process of modification of membrane lipids in response to environmental stimuli [1]. Dissertation investigations applied this concept to prokaryotic and eukaryotic organisms, and expanded the perception of environmental factors from exogenous organic solvents to intracellular environment. The field of lipidomics deals with the analysis of phospholipid and fatty acid components of membranes the changes that occur due to environmental stimuli and their biological significance [2-6]. The high sensitivity of mass spectrometry (MS) is an ideal tool for lipidomics allowing detection, quantification and structural elucidation [6]. Coupling of a mass spectrometer to a chromatographic system, such as gas chromatograph (GC), allows the separation of fatty acid methyl esters analytes prior to analysis [7]. The research investigations that comprise this dissertation are divided into three interrelated projects. The first project involved the analysis of composition and structure of Clostridium thermocellum membranes from wild-type and ethanol-adapted strains in response to adaptation of cultures to growth in ethanol. The hypothesis being that adaptation of cultures to growth in ethanol would result in compensatory change to the membrane composition. Rat mitochondrial fatty acid profiles isolated from brain, liver, kidney and heart tissues were compared. The hypothesis being that differences in cellular environments found among various tissues would be reflected in the mitochondrial membrane composition. These data support the concept that variations to the lipid content of neurological mitochondria may increase susceptibility to the products of oxidative stress. Lastly, changes in neurological mitochondria as a function of Alzheimer’s disease progression were studied. The hypothesis being that changes to the mitochondrial lipidome would be significantly reflected during advanced stages of AD, in addition to being more prevalent in regions displaying greater pathology. The three interrelated projects increased our understanding of the boundaries established by the concept of homeoviscous adaptation. Project specific hypotheses were supported by data obtained from these investigations

Automated robotic liquid handling assembly of modular DNA devices

Recent advances in modular DNA assembly techniques have enabled synthetic biologists to test significantly more of the available "design space" represented by "devices" created as combinations of individual genetic components. However, manual assembly of such large numbers of devices is time-intensive, error-prone, and costly. The increasing sophistication and scale of synthetic biology research necessitates an efficient, reproducible way to accommodate large-scale, complex, and high throughput device construction. Here, a DNA assembly protocol using the Type-IIS restriction endonuclease based Modular Cloning (MoClo) technique is automated on two liquid-handling robotic platforms. Automated liquid-handling robots require careful, often times tedious optimization of pipetting parameters for liquids of different viscosities (e.g. enzymes, DNA, water, buffers), as well as explicit programming to ensure correct aspiration and dispensing of DNA parts and reagents. This makes manual script writing for complex assemblies just as problematic as manual DNA assembly, and necessitates a software tool that can automate script generation. To this end, we have developed a web-based software tool, http://mocloassembly.com, for generating combinatorial DNA device libraries from basic DNA parts uploaded as Genbank files. We provide access to the tool, and an export file from our liquid handler software which includes optimized liquid classes, labware parameters, and deck layout. All DNA parts used are available through Addgene, and their digital maps can be accessed via the Boston University BDC ICE Registry. Together, these elements provide a foundation for other organizations to automate modular cloning experiments and similar protocols. The automated DNA assembly workflow presented here enables the repeatable, automated, high-throughput production of DNA devices, and reduces the risk of human error arising from repetitive manual pipetting. Sequencing data show the automated DNA assembly reactions generated from this workflow are ~95% correct and require as little as 4% as much hands-on time, compared to manual reaction preparation

Cognitive Flexibility Hypertext as a Learning Environment in Economics: A Pedagogical Note

Instructional design in complex subjects requires the application of a sophisticated theory of cognition.* Instructional techniques and strategies that work at the knowledge and comprehension stages of cognitive development may actually inhibit learning at more advanced levels. Evaluation and synthesis require a different cognitive paradigm. The theory of cognitive flexibility is a case-based approach for the development of upper-level cognitive skills, particularly the ability to transfer knowledge to novel situations. This paper applies the theory of cognitive flexibility to instructional design in economics. An example from monetary economics illustrates the adaptation of cognitive flexibility hypertext to a knowledge management interface

association of a large vulvar tumor in a in a young premenopausal woman with recurrent pseudoangiomatous stromal hyperplasia of the breasts

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Hot Rocks: Constraining the Thermal Conditions of the Mistastin Lake Impact Melt Deposits Using Zircon Grain Microstructures

The production of superheated melt during hypervelocity impact events has been proposed to be a common occurrence on terrestrial planetary bodies. Recent direct evidence of superheated impact melt temperatures exceeding \u3e2370°C from the Kamestastin (Mistastin Lake) impact structure, Canada, was based on a single impact glass sample. Such high superheated melt temperatures have strong implications for the evolution of crustal material, the thermal history of impact cratering events, and the rheology of impact melt. However, although widely predicted in previous studies, with the exception of the Mistastin Lake impact glass, there is little direct evidence for superheated temperatures in multiple settings across an impact structure. Therefore, an outstanding question is how heterogeneous are superheated conditions across a single impact structure. In this work, we analyze the crystallographic orientations and microstructures of zircon grains and the precursor parent phases of baddeleyite crystals, from four different samples representing the entire melt-bearing stratigraphy at Mistastin: an impact glass, a vesicular clast-poor impact melt rock, a clast-rich impact melt rock, and a glass-bearing impact breccia. Using electron microprobe analysis followed by electron backscatter diffraction, we discovered that four zircon grains with vermicular coronae of baddeleyite crystals from the impact glass contain evidence for a cubic zirconia precursor, indicative of temperature conditions \u3e2370°C. We also report evidence of superheating up to 1673°C in the glass-bearing impact breccia. In addition, we also report the first occurrence at Mistastin of the high-pressure zircon polymorph reidite and former reidite in granular neoblastic (FRIGN) zircon in grains from the glass-bearing impact breccia, implying minimum peak shocks from 30–40 GPa. The identification of superheating from two localities at Mistastin demonstrates (1) that superheating is not restricted solely to rapidly cooled impact melt rock samples and is therefore more distributed across impact structures, and (2) we can investigate the P-T evolution pathways of impact melt from different impact settings, providing a clearer picture of the thermal conditions and history of the impact structure

The Role of Perceived University Support in the Formation of Students' Entrepreneurial Intention

Entrepreneurship education is central to student entrepreneurship. Previous research has attempted to understand the role of entrepreneurship education in the formation of students' entrepreneurial intention and behavior, albeit in an isolated manner. Universities can support entrepreneurship in many ways, but it is important to measure students' perception of the support that they receive in order to understand the extent of such support and its impact on students. The current study proposed and tested an integrative, multiperspective framework. We have hypothesized that the three dimensions of university support, that is, perceived educational support, concept development support, and business development support, together with institutional support, shape students' entrepreneurial self-efficacy. In turn, entrepreneurial self-efficacy and individual motivations constitute the fundamental elements of the intention to start a business. A sample of 805 university students took part in the study and data were analyzed using structural equation modeling. Our findings showed that perceived educational support exerted the highest influence on entrepreneurial self-efficacy, followed by concept development support, business development support, and institutional support. Self-efficacy in turn had a significant effect on entrepreneurial intention. Individual motivations such as self-realization, recognition, and role had an additional impact on intention. However, intention was not related to financial success, innovation, and independence. The findings suggest that a holistic perspective provides a more meaningful understanding of the role of perceived university support in the formation of students' entrepreneurial intention. Theoretical and practical implications are discussed

Multi-wavelength lens construction of a Planck and Herschel-detected star-bursting galaxy

We present a source-plane reconstruction of a Herschel and Planck-detected gravitationally lensed dusty star-forming galaxy (DSFG) at z = 1.68 using Hubble, Submillimeter Array (SMA), and Keck observations. The background submillimeter galaxy (SMG) is strongly lensed by a foreground galaxy cluster at z = 0.997 and appears as an arc with a length of ∼15″ in the optical images. The continuum dust emission, as seen by SMA, is limited to a single knot within this arc. We present a lens model with source-plane reconstructions at several wavelengths to show the difference in magnification between the stars and dust, and highlight the importance of multi-wavelength lens models for studies involving lensed DSFGs. We estimate the physical properties of the galaxy by fitting the flux densities to model spectral energy distributions leading to a magnification-corrected starformation rate (SFR) of 390 ± 60 M yr−1 and a stellar mass of 1.1 ± 0.4 10 x 11 M. These values are consistent with high-redshift massive galaxies that have formed most of their stars already. The estimated gas-to-baryon fraction, molecular gas surface density, and SFR surface density have values of 0.43 ± 0.13, 350 ± 200 M pc−2, and ~ 12 7 M yr−1 kpc−2, respectively. The ratio of SFR surface density to molecular gas surface density puts this among the most star-forming systems, similar to other measured SMGs and local ULIRGs

A Survey of New Temperature-Sensitive, Embryonic-Lethal Mutations in C. elegans: 24 Alleles of Thirteen Genes

To study essential maternal gene requirements in the early C. elegans embryo, we have screened for temperature-sensitive, embryonic lethal mutations in an effort to bypass essential zygotic requirements for such genes during larval and adult germline development. With conditional alleles, multiple essential requirements can be examined by shifting at different times from the permissive temperature of 15°C to the restrictive temperature of 26°C. Here we describe 24 conditional mutations that affect 13 different loci and report the identity of the gene mutations responsible for the conditional lethality in 22 of the mutants. All but four are mis-sense mutations, with two mutations affecting splice sites, another creating an in-frame deletion, and one creating a premature stop codon. Almost all of the mis-sense mutations affect residues conserved in orthologs, and thus may be useful for engineering conditional mutations in other organisms. We find that 62% of the mutants display additional phenotypes when shifted to the restrictive temperature as L1 larvae, in addition to causing embryonic lethality after L4 upshifts. Remarkably, we also found that 13 out of the 24 mutations appear to be fast-acting, making them particularly useful for careful dissection of multiple essential requirements. Our findings highlight the value of C. elegans for identifying useful temperature-sensitive mutations in essential genes, and provide new insights into the requirements for some of the affected loci