In situ synthesis of DNA microarray on functionalized cyclic olefin copolymer substrate.

Thermoplastic materials such as cyclic-olefin copolymers (COC) provide a versatile and cost-effective alternative to the traditional glass or silicon substrate for rapid prototyping and industrial scale fabrication of microdevices. To extend the utility of COC as an effective microarray substrate, we developed a new method that enabled for the first time in situ synthesis of DNA oligonucleotide microarrays on the COC substrate. To achieve high-quality DNA synthesis, a SiO(2) thin film array was prepatterned on the inert and hydrophobic COC surface using RF sputtering technique. The subsequent in situ DNA synthesis was confined to the surface of the prepatterned hydrophilic SiO(2) thin film features by precision delivery of the phosphoramidite chemistry using an inkjet DNA synthesizer. The in situ SiO(2)-COC DNA microarray demonstrated superior quality and stability in hybridization assays and thermal cycling reactions. Furthermore, we demonstrate that pools of high-quality mixed-oligos could be cleaved off the SiO(2)-COC microarrays and used directly for construction of DNA origami nanostructures. It is believed that this method will not only enable synthesis of high-quality and low-cost COC DNA microarrays but also provide a basis for further development of integrated microfluidics microarrays for a broad range of bioanalytical and biofabrication applications.

"Double Rainbow," "Appalachiana," and "'The Invisible Mass': Exploring Compositional Technique in Alfred Schnittke's Second Symphony"

This dissertation consists of three distinct components: (1) “Double Rainbow,” a notated composition for an acoustic ensemble of 10 instruments, ca. 36 minutes. (2) “Appalachiana”, a fixed-media composition for electro-acoustic music and video, ca. 30 minutes, and (3) “'The Invisible Mass': Exploring Compositional Technique in Alfred Schnittke’s Second Symphony”, an analytical article.

(1) Double Rainbow is a ca. 36 minute composition in four movements scored for 10 instruments: flute, Bb clarinet (doubling on bass clarinet), tenor saxophone (doubling on alto saxophone), french horn, percussion (glockenspiel, vibraphone, wood block, 3 toms, snare drum, bass drum, suspended cymbal), piano, violin, viola, cello, and double bass. Each of the four movements of the piece explore their own distinct character and set of compositional goals. The piece is presented as a musical score and as a recording, which was extensively treated in post-production.

(2) Appalachiana, is a ca. 30 minute fixed-media composition for music and video. The musical component was created as a vehicle to showcase several approaches to electro-acoustic music composition –fft re-synthesis for time manipulation effects, the use of a custom-built software instrument which implements generative approaches to creating rhythm and pitch patterns, using a recording of rain to create rhythmic triggers for software instruments, and recording additional components with acoustic instruments. The video component transforms footage of natural landscapes filmed at several locations in North Carolina, Virginia, and West Virginia into a surreal narrative using a variety of color, lighting, distortion, and time-manipulation video effects.

(3) “‘The Invisible Mass:’ Exploring Compositional Technique in Alfred Schnittke’s Second Symphony” is an analytical article that focuses on Alfred Schnittke’s compositional technique as evidenced in the construction of his Second Symphony and discussed by the composer in a number of previously untranslated articles and interviews. Though this symphony is pivotal in the composer’s oeuvre, there are currently no scholarly articles that offer in-depth analyses of the piece. The article combines analyses of the harmony, form, and orchestration in the Second Symphony with relevant quotations from the composer, some from published and translated sources and others newly translated by the author from research at the Russian State Library in St. Petersburg. These offer a perspective on how Schnittke’s compositional technique combines systematic geometric design with keen musical intuition.

The Synthesis of Novel N-Heterocyclic Scaffolds and Diazirine-Based Molecular Tags

N-Heterocycles are ubiquitous in biologically active natural products and pharmaceuticals. Yet, new syntheses and modifications of N-heterocycles are continually of interest for the purposes of expanding chemical space, finding quicker synthetic routes, better pharmaceuticals, and even new handles for molecular labeling. There are several iterations of molecular labeling; the decision of where to place the label is as important as of which visualization technique to emphasize.

Piperidine and indole are two of the most widely distributed N-heterocycles and thus were targeted for synthesis, functionalization, and labeling. The major functionalization of these scaffolds should include a nitrogen atom, while the inclusion of other groups will expand the utility of the method. Towards this goal, ease of synthesis and elimination of step-wise transformations are of the utmost concern. Here, the concept of electrophilic amination can be utilized as a way of introducing complex secondary and tertiary amines with minimal operations.

Molecular tags should be on or adjacent to an N-heterocycle as they are normally the motifs implicated at the binding site of enzymes and receptors. The labeling techniques should be useful to a chemical biologist, but should also in theory be useful to the medical community. The two types of labeling that are of interest to a chemist and a physician would be positron emission tomography (PET) and magnetic resonance imaging (MRI).

Coincidentally, the 3-positions of both piperidine and indole are historically difficult to access and modify. However, using electrophilic amination techniques, 3-functionalized piperidines can be synthesized in good yields from unsaturated amines. In the same manner, 3-labeled piperidines can be obtained; the piperidines can either be labeled with an azide for biochemical research or an 18F for PET imaging research. The novel electrophiles, N-benzenesulfonyloxyamides, can be reacted with indole in one of two ways: 3-amidation or 1-amidomethylation, depending on the exact reaction conditions. Lastly, a novel, hyperpolarizable 15N2-labeled diazirine has been developed as an exogenous and versatile tag for use in magnetic resonance imaging.