DNA-Mediated Charge Transport Devices for Protein Detection

Detection of biologically relevant targets, including small molecules, proteins, DNA, and RNA, is vital for fundamental research as well as clinical diagnostics. Sensors with biological elements provide a natural foundation for such devices because of the inherent recognition capabilities of biomolecules. Electrochemical DNA platforms are simple, sensitive, and do not require complex target labeling or expensive instrumentation. Sensitivity and specificity are added to DNA electrochemical platforms when the physical properties of DNA are harnessed. The inherent structure of DNA, with its stacked core of aromatic bases, enables DNA to act as a wire via DNA-mediated charge transport (DNA CT). DNA CT is not only robust over long molecular distances of at least 34 nm, but is also especially sensitive to anything that perturbs proper base stacking, including DNA mismatches, lesions, or DNA-binding proteins that distort the π-stack. Electrochemical sensors based on DNA CT have previously been used for single-nucleotide polymorphism detection, hybridization assays, and DNA-binding protein detection. Here, improvements to (i) the structure of DNA monolayers and (ii) the signal amplification with DNA CT platforms for improved sensitivity and detection are described.

First, improvements to the control over DNA monolayer formation are reported through the incorporation of copper-free click chemistry into DNA monolayer assembly. As opposed to conventional film formation involving the self-assembly of thiolated DNA, copper-free click chemistry enables DNA to be tethered to a pre-formed mixed alkylthiol monolayer. The total amount of DNA in the final film is directly related to the amount of azide in the underlying alkylthiol monolayer. DNA monolayers formed with this technique are significantly more homogeneous and lower density, with a larger amount of individual helices exposed to the analyte solution. With these improved monolayers, significantly more sensitive detection of the transcription factor TATA binding protein (TBP) is achieved.

Using low-density DNA monolayers, two-electrode DNA arrays were designed and fabricated to enable the placement of multiple DNA sequences onto a single underlying electrode. To pattern DNA onto the primary electrode surface of these arrays, a copper precatalyst for click chemistry was electrochemically activated at the secondary electrode. The location of the secondary electrode relative to the primary electrode enabled the patterning of up to four sequences of DNA onto a single electrode surface. As opposed to conventional electrochemical readout from the primary, DNA-modified electrode, a secondary microelectrode, coupled with electrocatalytic signal amplification, enables more sensitive detection with spatial resolution on the DNA array electrode surface. Using this two-electrode platform, arrays have been formed that facilitate differentiation between well-matched and mismatched sequences, detection of transcription factors, and sequence-selective DNA hybridization, all with the incorporation of internal controls.

For effective clinical detection, the two working electrode platform was multiplexed to contain two complementary arrays, each with fifteen electrodes. This platform, coupled with low density DNA monolayers and electrocatalysis with readout from a secondary electrode, enabled even more sensitive detection from especially small volumes (4 μL per well). This multiplexed platform has enabled the simultaneous detection of two transcription factors, TBP and CopG, with surface dissociation constants comparable to their solution dissociation constants.

With the sensitivity and selectivity obtained from the multiplexed, two working electrode array, an electrochemical signal-on assay for activity of the human methyltransferase DNMT1 was incorporated. DNMT1 is the most abundant human methyltransferase, and its aberrant methylation has been linked to the development of cancer. However, current methods to monitor methyltransferase activity are either ineffective with crude samples or are impractical to develop for clinical applications due to a reliance on radioactivity. Electrochemical detection of methyltransferase activity, in contrast, circumvents these issues. The signal-on detection assay translates methylation events into electrochemical signals via a methylation-specific restriction enzyme. Using the two working electrode platform combined with this assay, DNMT1 activity from tumor and healthy adjacent tissue lysate were evaluated. Our electrochemical measurements revealed significant differences in methyltransferase activity between tumor tissue and healthy adjacent tissue.

As differential activity was observed between colorectal tumor tissue and healthy adjacent tissue, ten tumor sets were subsequently analyzed for DNMT1 activity both electrochemically and by tritium incorporation. These results were compared to expression levels of DNMT1, measured by qPCR, and total DNMT1 protein content, measured by Western blot. The only trend detected was that hyperactivity was observed in the tumor samples as compared to the healthy adjacent tissue when measured electrochemically. These advances in DNA CT-based platforms have propelled this class of sensors from the purely academic realm into the realm of clinically relevant detection.

Microelectrode Implants for Spinal Cord Stimulation in Rats

Paralysis is a debilitating condition afflicting millions of people across the globe, and is particularly deleterious to quality of life when motor function of the legs is severely impaired or completely absent. Fortunately, spinal cord stimulation has shown great potential for improving motor function after spinal cord injury and other pathological conditions. Many animal studies have shown stimulation of the neural networks in the spinal cord can improve motor ability so dramatically that the animals can even stand and step after a complete spinal cord transaction.

This thesis presents work to successfully provide a chronically implantable device for rats that greatly enhances the ability to control the site of spinal cord stimulation. This is achieved through the use of a parylene-C based microelectrode array, which enables a density of stimulation sites unattainable with conventional wire electrodes. While many microelectrode devices have been proposed in the past, the spinal cord is a particularly challenging environment due to the bending and movement it undergoes in a live animal. The developed microelectrode array is the first to have been implanted in vivo while retaining functionality for over a month. In doing so, different neural pathways can be selectively activated to facilitate standing and stepping in spinalized rats using various electrode combinations, and important differences in responses are observed.

An engineering challenge for the usability of any high density electrode array is connecting the numerous electrodes to a stimulation source. This thesis develops several technologies to address this challenge, beginning with a fully passive implant that uses one wire per electrode to connect to an external stimulation source. The number of wires passing through the body and the skin proved to be a hazard for the health of the animal, so a multiplexed implant was devised in which active electronics reduce the number of wires. Finally, a fully wireless implant was developed. As these implants are tested in vivo, encapsulation is of critical importance to retain functionality in a chronic experiment, especially for the active implants, and it was achieved without the use of costly ceramic or metallic hermetic packaging. Active implants were built that retained functionality 8 weeks after implantation, and achieved stepping in spinalized rats after just 8-10 days, which is far sooner than wire-based electrical stimulation has achieved in prior work.

Toward Understanding Astrophysical Phenomena

Fast radio bursts (FRBs), a novel type of radio pulse, whose physics is not yet understood at all. Only a handful of FRBs had been detected when we started this project. Taking account of the scant observations, we put physical constraints on FRBs. We excluded proposals of a galactic origin for their extraordinarily high dispersion measures (DM), in particular stellar coronas and HII regions. Therefore our work supports an extragalactic origin for FRBs. We show that the resolved scattering tail of FRB 110220 is unlikely to be due to propagation through the intergalactic plasma. Instead the scattering is probably caused by the interstellar medium in the FRB's host galaxy, and indicates that this burst sits in the central region of that galaxy. Pulse durations of order $\ms$ constrain source sizes of FRBs implying enormous brightness temperatures and thus coherent emission. Electric fields near FRBs at cosmological distances would be so strong that they could accelerate free electrons from rest to relativistic energies in a single wave period. When we worked on FRBs, it was unclear whether they were genuine astronomical signals as distinct from `perytons', clearly terrestrial radio bursts, sharing some common properties with FRBs. Recently, in April 2015, astronomers discovered that perytons were emitted by microwave ovens. Radio chirps similar to FRBs were emitted when their doors opened while they were still heating. Evidence for the astronomical nature of FRBs has strengthened since our paper was published. Some bursts have been found to show linear and circular polarizations and Faraday rotation of the linear polarization has also been detected. I hope to resume working on FRBs in the near future. But after we completed our FRB paper, I decided to pause this project because of the lack of observational constraints.

The pulsar triple system, J0733+1715, has its orbital parameters fitted to high accuracy owing to the precise timing of the central $\ms$ pulsar. The two orbits are highly hierarchical, namely $P_{\mathrm{orb,1}}\ll P_{\mathrm{orb,2}}$, where 1 and 2 label the inner and outer white dwarf (WD) companions respectively. Moreover, their orbital planes almost coincide, providing a unique opportunity to study secular interaction associated purely with eccentricity beyond the solar system. Secular interaction only involves effect averaged over many orbits. Thus each companion can be represented by an elliptical wire with its mass distributed inversely proportional to its local orbital speed. Generally there exists a mutual torque, which vanishes only when their apsidal lines are parallel or anti-parallel. To maintain either mode, the eccentricity ratio, $e_1/e_2$, must be of the proper value, so that both apsidal lines precess together. For J0733+1715, $e_1\ll e_2$ for the parallel mode, while $e_1\gg e_2$ for the anti-parallel one. We show that the former precesses $\sim 10$ times slower than the latter. Currently the system is dominated by the parallel mode. Although only a little anti-parallel mode survives, both eccentricities especially $e_1$ oscillate on $\sim 10^3\yr$ timescale. Detectable changes would occur within $\sim 1\yr$. We demonstrate that the anti-parallel mode gets damped $\sim 10^4$ times faster than its parallel brother by any dissipative process diminishing $e_1$. If it is the tidal damping in the inner WD, we proceed to estimate its tidal quantity parameter ($Q$) to be $\sim 10^6$, which was poorly constrained by observations. However, tidal damping may also happen during the preceding low-mass X-ray binary (LMXB) phase or hydrogen thermal nuclear flashes. But, in both cases, the inner companion fills its Roche lobe and probably suffers mass/angular momentum loss, which might cause $e_1$ to grow rather than decay.

Several pairs of solar system satellites occupy mean motion resonances (MMRs). We divide these into two groups according to their proximity to exact resonance. Proximity is measured by the existence of a separatrix in phase space. MMRs between Io-Europa, Europa-Ganymede and Enceladus-Dione are too distant from exact resonance for a separatrix to appear. A separatrix is present only in the phase spaces of the Mimas-Tethys and Titan-Hyperion MMRs and their resonant arguments are the only ones to exhibit substantial librations. When a separatrix is present, tidal damping of eccentricity or inclination excites overstable librations that can lead to passage through resonance on the damping timescale. However, after investigation, we conclude that the librations in the Mimas-Tethys and Titan-Hyperion MMRs are fossils and do not result from overstability.

Rubble piles are common in the solar system. Monolithic elements touch their neighbors in small localized areas. Voids occupy a significant fraction of the volume. In a fluid-free environment, heat cannot conduct through voids; only radiation can transfer energy across them. We model the effective thermal conductivity of a rubble pile and show that it is proportional the square root of the pressure, $P$, for $P\leq \epsy^3\mu$ where $\epsy$ is the material's yield strain and $\mu$ its shear modulus. Our model provides an excellent fit to the depth dependence of the thermal conductivity in the top $140\,\mathrm{cm}$ of the lunar regolith. It also offers an explanation for the low thermal inertias of rocky asteroids and icy satellites. Lastly, we discuss how rubble piles slow down the cooling of small bodies such as asteroids.

Electromagnetic (EM) follow-up observations of gravitational wave (GW) events will help shed light on the nature of the sources, and more can be learned if the EM follow-ups can start as soon as the GW event becomes observable. In this paper, we propose a computationally efficient time-domain algorithm capable of detecting gravitational waves (GWs) from coalescing binaries of compact objects with nearly zero time delay. In case when the signal is strong enough, our algorithm also has the flexibility to trigger EM observation {\it before} the merger. The key to the efficiency of our algorithm arises from the use of chains of so-called Infinite Impulse Response (IIR) filters, which filter time-series data recursively. Computational cost is further reduced by a template interpolation technique that requires filtering to be done only for a much coarser template bank than otherwise required to sufficiently recover optimal signal-to-noise ratio. Towards future detectors with sensitivity extending to lower frequencies, our algorithm's computational cost is shown to increase rather insignificantly compared to the conventional time-domain correlation method. Moreover, at latencies of less than hundreds to thousands of seconds, this method is expected to be computationally more efficient than the straightforward frequency-domain method.

Design of an aerial tramway

An Aerial Tramway is a device for transporting material in receptacles over wire ropes supported at various elevations above the ground by means of posts or standards. Wire rope has been known for many centuries, and there is a drawing of a ropeway appearing in a book dated 1411.

Mean-speed measurements in two-dimensional, incompressible, fully-developed turbulent channel flow

Mean velocity profiles were measured in the 5” x 60” wind channel of the turbulence laboratory at the GALCIT, by the use of a hot-wire anemometer. The repeatability of results was established, and the accuracy of the instrumentation estimated. Scatter of experimental results is a little, if any, beyond this limit, although some effects might be expected to arise from variations in atmospheric humidity, no account of this factor having been taken in the present work. Also, slight unsteadiness in flow conditions will be responsible for some scatter.

Irregularities of a hot-wire in close proximity to a solid boundary at low speeds were observed, as have already been found by others.

That Kármán’s logarithmic law holds reasonably well over the main part of a fully developed turbulent flow was checked, the equation u/u_t = 6.0 + 6.25 log₁₀ y^ut/v being obtained, and, as has been previously the case, the experimental points do not quite form one straight line in the region where viscosity effects are small. The values of the constants for this law for the best over-all agreement were determined and compared with those obtained by others.

The range of Reynolds numbers used (based on half-width of channel) was from 20,000 to 60,000.

Heteroepitaxy of Group IV and Group III-V semiconductor alloys for photovoltaic applications

Photovoltaic energy conversion represents a economically viable technology for realizing collection of the largest energy resource known to the Earth -- the sun. Energy conversion efficiency is the most leveraging factor in the price of energy derived from this process. This thesis focuses on two routes for high efficiency, low cost devices: first, to use Group IV semiconductor alloy wire array bottom cells and epitaxially grown Group III-V compound semiconductor alloy top cells in a tandem configuration, and second, GaP growth on planar Si for heterojunction and tandem cell applications.

Metal catalyzed vapor-liquid-solid grown microwire arrays are an intriguing alternative for wafer-free Si and SiGe materials which can be removed as flexible membranes. Selected area Cu-catalyzed vapor-liquid solid growth of SiGe microwires is achieved using chlorosilane and chlorogermane precursors. The composition can be tuned up to 12% Ge with a simultaneous decrease in the growth rate from 7 to 1 μm/min^-1. Significant changes to the morphology were observed, including tapering and faceting on the sidewalls and along the lengths of the wires. Characterization of axial and radial cross sections with transmission electron microscopy revealed no evidence of defects at facet corners and edges, and the tapering is shown to be due to in-situ removal of catalyst material during growth. X-ray diffraction and transmission electron microscopy reveal a Ge-rich crystal at the tip of the wires, strongly suggesting that the Ge incorporation is limited by the crystallization rate.

Tandem Ga_1-xIn_xP/Si microwire array solar cells are a route towards a high efficiency, low cost, flexible, wafer-free solar technology. Realizing tandem Group III-V compound semiconductor/Si wire array devices requires optimization of materials growth and device performance. GaP and Ga_1-xIn_xP layers were grown heteroepitaxially with metalorganic chemical vapor deposition on Si microwire array substrates. The layer morphology and crystalline quality have been studied with scanning electron microscopy and transmission electron microscopy, and they provide a baseline for the growth and characterization of a full device stack. Ultimately, the complexity of the substrates and the prevalence of defects resulted in material without detectable photoluminescence, unsuitable for optoelectronic applications.

Coupled full-field optical and device physics simulations of a Ga_0.51In_0.49P/Si wire array tandem are used to predict device performance. A 500 nm thick, highly doped "buffer" layer between the bottom cell and tunnel junction is assumed to harbor a high density of lattice mismatch and heteroepitaxial defects. Under simulated AM1.5G illumination, the device structure explored in this work has a simulated efficiency of 23.84% with realistic top cell SRH lifetimes and surface recombination velocities. The relative insensitivity to surface recombination is likely due to optical generation further away from the free surfaces and interfaces of the device structure.

Finally, GaP has been grown free of antiphase domains on Si (112) oriented substrates using metalorganic chemical vapor deposition. Low temperature pulsed nucleation is followed by high temperature continuous growth, yielding smooth, specular thin films. Atomic force microscopy topography mapping showed very smooth surfaces (4-6 Å RMS roughness) with small depressions in the surface. Thin films (~ 50 nm) were pseudomorphic, as confirmed by high resolution x-ray diffraction reciprocal space mapping, and 200 nm thick films showed full relaxation. Transmission electron microscopy showed no evidence of antiphase domain formation, but there is a population of microtwin and stacking fault defects.

Análisis de la fabricación de las tijas de una motocicleta de competición. Prototipo

[ES]Este trabajo fin de grado, TFG en adelante, consiste en el análisis de la fabricación de las tijas de una motocicleta de competición Motostudent. Se trata de conseguir fabricar las tijas de una manera óptima, tanto en que su material como el proceso de fabricación sea el más adecuado para su función siendo, además, lo más económico posible. Para llevar a cabo este trabajo es necesario conocer las cualidades que una tija debe cubrir en una motocicleta de competición. A partir de dichas cualidades se selecciona el material idóneo de entre otros muchos, estudiando las propiedades fiscas, químicas y mecánicas de cada uno de ellos. También se hará un estudio de dos alternativas de fabricación posibles, electroerosión y arranque de viruta con herramienta, para a posteriori elegir la más indicada. La idea no es construir un diseño final para la fabricación, sino un prototipo sobre el cual hacer las pruebas oportunas y posteriormente realizar cambios y mejoras para posteriores ediciones. El presente proyecto marcará las pautas de diseño y el proceso a seguir durante la fabricación de las tijas que el equipo representante de la Escuela técnica superior de Bilbao, ETSIB en adelante, empleará en la futura competición.

Photodisintegration of helium-3 at energies between 200 and 600 MEV

We have measured differential cross-sections for the two-body photodisintegration of Helium-3, ɣ + He³ → p + d, between incident photon energies of 200 and 600 MeV, and for center of mass frame angles between 30° and 150°. Both final state particles were detected in arrays of wire spark chambers and scintillation counters; the high momentum particle was analyzed in a magnet spectrometer. The results are interpreted in terms of amplitudes to produce the ∆(1236) resonance in an intermediate state, as well as non-resonant amplitudes. This experiment, together with an (unfinished) experiment on the inverse reaction, p + d → He³ + ɣ, will provide a reciprocity test of time reversal invariance.

五自由度纳米级定位精度机构的研制

提出一种新型的五自由度精密定位平台的工作原理及其设计方法。工作台采用压电陶瓷作为驱动元件，柔性导向机构实现平移及转动功能。整个工作台可由整块金属材料通过线切割加工制成，实现一体化加工，而且结构紧凑。并给出导向机构刚度计算公式及设计实例。

位相锁定来自同一侧面抽运Nd：YAG板条的两束激光

实验研究了腔内位相锁定来至一LAD侧面抽运的Nd：YAG板条的两束激光，输出镜面出现干涉条纹，获得1.13W的相干光，其组束效率达到64.9%，相干度约60%。实验中发现只需要一根作为滤波的金属丝放在离输出镜合适的位置都就能有效稳定干涉条纹，金属丝引起的损耗低于8%。

A travessia de Nanetto Pipetta e o imaginário dos imigrantes italianos para o Sul do Brasil, em busca do país da Cocanha

Esta dissertação aborda o tema da imigração italiana à luz do imaginário daqueles que se dirigiam em massa para o Brasil, entre o final do século XIX e início do XX, em busca de um mundo novo, perseguindo o sonho de um Paraíso Terrestre, o país imaginário da Cocanha. O tema é abordado a partir de obras literárias, como o poema francês, datado do século XIII, o Fabliau da Cocanha e da narrativa Vita e Stòria di Nanetto Pipetta, que apresenta o jovem personagem imigrante clandestino vêneto, criado por Aquiles Bernardi, e cujas aventuras eram publicadas semanalmente em capítulos no jornal gaúcho Stafetta Riograndense, nos anos 1924-1925. A trama vivida pelo personagem que dá título à obra é fio condutor das reflexões apresentadas. Para o imigrante italiano, o imaginário sobre o Brasil como o país da Cocanha encontra neste personagem sua expressão fiel, mantendo acesa a chama da esperança de uma vida melhor para si e para as gerações futuras.

Comparison of six tag types in sea-trout tagging experiments in the Baltic Sea

In the international Baltic Sea trout tagging experiment 27 753 hatchery reared sea trout smolts were tagged in Denmark, Finland, Poland and Sweden in 1979 and 1980. The fish were tagged with the original Carlin tag, two modified Carlin tag types (Canadian and Finnish polythene), streamer and Floy tags and Polish tags attached with Monel metal wire. The tag returns were affected by the place of release and smolt quality. The best results were obtained in the case of tags attached with double wire or thread -original Carlin, Canadian and Finnish polythene. The poorest results were obtained with streamer tags.

A comparison between circle hook and J hook performance in the dolphinfish, yellowfish tuna, and wahoo troll fishery off the coast of North Carolina

We compared numbers of strikes, proportions of fish that hooked up after strikes, proportions of fish that stayed on hook (retained) after hook up, and numbers of fish caught between circle and J hooks rigged with dead natural fish bait (ballyhoo)and trolled for three oceanic predator species: dolphinfish (Coryphaena hippurus), yellowfin tuna (Thunnus albacares), and wahoo (Acanthocybium solandri). Interactions were compared between circle and J hooks fished on 75 trips by two user groups (charter and recreational fishermen). Hooks were affixed to three species-specific leader types most commonly fished in this region: monofilament (dolphinfish), fluorocarbon (tuna), and wire (wahoo). Numbers of fish caught per trip and three potential mechanisms that might inf luence numbers caught (i.e., number of strikes, proportion of fish hooked, and proportion retained) were modeled with generalized linear models that considered hook type, leader type, species, user (fishing) group, and wave height as main effects. Hook type was a main effect at the catch level; generally, more fish were caught on J hooks than on circle hooks. The effect of hook type on strike rates was equivocal. However, J hooks had a greater proportion of hook-ups than did circle hooks. Finally, the proportion of fish retained once hooked was generally equal between hook types. We found similar results when data from additional species were pooled as a “tuna” group and a “mackerel” group. We conclude that J hooks are more effective than circle hooks at the hook-up level and result in greater numbers of troll-caught dolphinfish, tunas

A materialidade e o adverso nos Bólides de Hélio Oiticica

A pesquisa analisa a criação de adversidade através da materialidade crua e rústica utilizada nos Bólides de Hélio Oiticica. O papel da materialidade tornou-se evidente a partir da percepção de que os ditos objetos realizam um convite àquilo que chamamos de participação adversa. Ao mesmo tempo em que eram objetos capazes de despertar a curiosidade do espectador e incitá-lo à manipulação, o faziam através do toque de substâncias ásperas e pouco suaves, como por exemplo, pedaços de madeira mal pintada, telas de nylon, conchas e pigmento em pó. O despertar de sensações contrárias ao prazer sensorial sugeriu que não apenas a materialidade estaria no cerne da questão da criação de adversidade como um dos fios condutores da obra de Oiticica, como também que a captura de objetos descartados oriundos do cotidiano seria a maneira do artista estruturar a sua noção de adversidade (da vida e da arte). Para compreender o papel estrutural dos objetos despejados do dia-a-dia, realizamos a aproximação entre o adverso e o abjeto (KRAUSS, 2000), bem como o pensamento do informe como elemento operacional da obra (YVE-ALAIN BOIS, 2000). A materialidade rude foi vista ainda como criadora de uma temporalidade de instantes para os Bólides e aqueles que os manipulavam, como organizadora de fricção com o espaço e a cultura e ainda como elemento de rebaixamento da condição matérica e do espectador

Seasonal changes in growth of coho salmon (Oncorhynchus kisutch) off Oregon and Washington and concurrent changes in the spacing of scale circuli

In this study we present new information on seasonal variation in absolute growth rate in length of coho salmon (Oncorhynchus kisutch) in the ocean off Oregon and Washington, and relate these changes in growth rate to concurrent changes in the spacing of scale circuli. Average spacing of scale circuli and average rate of circulus formation were significantly and positively correlated with average growth rate among groups of juvenile and maturing coho salmon and thus could provide estimates of growth between age groups and seasons. Regression analyses indicated that the spacing of circuli was proportional to the scale growth rate raised to the 0.4−0.6 power. Seasonal changes in the spacing of scale circuli reflected seasonal changes in apparent growth rates of fish. Spacing of circuli at the scale margin was greatest during the spring and early summer, decreased during the summer, and was lowest in winter or early spring. Changes over time in length of fish caught during research cruises indicated that the average growth rate of juvenile coho salmon between June and September was about 1.3 mm/d and then decreased during the fall and winter to about 0.6 mm/d. Average growth rate of maturing fish was about 2 mm/d between May and June, then decreased to about 1 mm/d between June and September. Average apparent growth rates of groups of maturing coded-wire−tagged coho salmon caught in the ocean hook-and-line fisheries also decreased between June and September. Our results indicate that seasonal change in the spacing of scale circuli is a useful indicator of seasonal change in growth rate of coho salmon in the ocean.