Mimicking the Arterial Microenvironment with PEG-PC to Investigate the Roles of Physicochemical Stimuli in SMC Phenotype and Behavior

The goal of this dissertation was to parse the roles of physical, mechanical and chemical cues in the phenotype plasticity of smooth muscle cells (SMCs) in atherosclerosis. We first developed and characterized a novel synthetic hydrogel with desirable traits for studying mechanotransduction in vitro. This hydrogel, PEG-PC, is a co-polymer of poly(ethylene glycol) and phosphorylcholine with an incredible range of Young’s moduli (~1 kPa - 9 MPa) that enables reproduction of nearly any tissue stiffness, exceptional optical and anti-fouling properties, and support for covalent attachment of extracellular matrix (ECM) proteins. To our knowledge, this combination of mechanical range, low price, and ease-of-use is unmatched by any other hydrogel. We further used PEG-PC to evaluate the impact of substrate stiffness on the proliferation and adhesion properties of three cancer cell lines in 2D, from which we conclude that mechanotransduction is cell type-dependent and differences in focal adhesion-mediated signaling affect proliferation outcomes. With PEG-PC as a substrate, we then designed a complex in vitro model to recapitulate characteristic changes in the surrounding microenvironment that SMCs experience during the progression of atherosclerosis. These changes include the composition of the ECM, the availability of soluble factors, and the surrounding mechanical environment. Our findings point to ECM composition as the primary regulator of SMC behaviors and characteristics, in part by modulating the effects of soluble factors. Unexpectedly, changes in substrate stiffness had a relatively modest effect. In spite of large ECM-directed differences in proliferation and motility, we did not find that these behaviors are inversely related to SMC marker expression, nor was marker expression substantially dependent on ECM composition despite being regulated by focal adhesion kinase signaling. Finally, our findings suggest that the transition from a migratory to a proliferative phenotype in atherosclerosis is mediated by the changing ECM composition, and we propose hypothetical, integrin-driven models to explain this switch. From these conclusions, we emphasize the importance of increasing the complexity of in vitro models to carefully match critical features of the in vivo microenvironments. We expect this approach to produce physiologically relevant behaviors, and in doing so we may identify novel, context-dependent therapeutic targets

Pyramidal Atoms: Berylliumlike Hollow States

Based on the idea that four excited electrons arrange themselves around the nucleus in the corners of a pyramid in order to minimize their mutual repulsion, we present an analytical model of quadruply excited states. The model shows excellent comparison with ab initio results and provides a clear physical picture of the intrinsic motion of the four electrons. The model is used to predict configuration-mixing fractions and spectra of these highly correlated states.Comment: 4 pages, 2 figure

High Altitude Ozone Research Balloon

In order to create a mission model of the high altitude ozone research balloon (HAORB) several options for flight preparation, altitude control, flight termination, and payload recovery were considered. After the optimal launch date and location for two separate HAORB flights were calculated, a method for reducing the heat transfer from solar and infrared radiation was designed and analytically tested. This provided the most important advantage of the HAORB over conventional balloons, i.e., its improved flight duration. Comparisons of different parachute configurations were made, and a design best suited for the HAORB's needs was determined to provide for payload recovery after flight termination. In an effort to avoid possible payload damage, a landing system was also developed

Telomere formation on macronuclear chromosomes of Oxytricha trifallax and O. fallax: alternatively processed regions have multiple telomere addition sites

BACKGROUND: Ciliates employ massive chromatid breakage and de novo telomere formation during generation of the somatic macronucleus. Positions flanking the 81-MAC locus are reproducibly cut. But those flanking the Common Region are proposed to often escape cutting, generating three nested macronuclear chromosomes, two retaining "arms" still appended to the Common Region. Arm-distal positions must differ (in cis) from the Common Region flanks. RESULTS: The Common-Region-flanking positions also differ from the arm-distal positions in that they are "multi-TAS" regions: anchored PCR shows heterogeneous patterns of telomere addition sites, but arm-distal sites do not. The multi-TAS patterns are reproducible, but are sensitive to the sequence of the allele being processed. Thus, random degradation following chromatid cutting does not create this heterogeneity; these telomere addition sites also must be dictated by cis-acting sequences. CONCLUSIONS: Most ciliates show such micro-heterogeneity in the precise positions of telomere addition sites. Telomerase is believed to be tightly associated with, and act in concert with, the chromatid-cutting nuclease: heterogeneity must be the result of intervening erosion activity. Our "weak-sites" hypothesis explains the correlation between alternative chromatid cutting at the Common Region boundaries and their multi-TAS character: when the chromatid-breakage machine encounters either a weak binding site or a weak cut site at these regions, then telomerase dissociates prematurely, leaving the new end subject to erosion by an exonuclease, which pauses at cis-acting sequences; telomerase eventually heals these resected termini. Finally, we observe TAS positioning influenced by trans-allelic interactions, reminiscent of transvection

A set-valued framework for birth-and-growth process

We propose a set-valued framework for the well-posedness of birth-and-growth process. Our birth-and-growth model is rigorously defined as a suitable combination, involving Minkowski sum and Aumann integral, of two very general set-valued processes representing nucleation and growth respectively. The simplicity of the used geometrical approach leads us to avoid problems arising by an analytical definition of the front growth such as boundary regularities. In this framework, growth is generally anisotropic and, according to a mesoscale point of view, it is not local, i.e. for a fixed time instant, growth is the same at each space point

Effects of Activation Drills Combined with Dynamic Warmup on Performance Field Tests

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Sorafenib resistance and JNK signaling in carcinoma during extracellular matrix stiffening

Tumor progression is coincident with mechanochemical changes in the extracellular matrix (ECM). We hypothesized that tumor stroma stiffening, alongside a shift in the ECM composition from a basement membrane-like microenvironment toward a dense network of collagen-rich fibers during tumorigenesis, confers resistance to otherwise powerful chemotherapeutics. To test this hypothesis, we created a high-throughput drug screening platform based on our poly(ethylene glycol)-phosphorylcholine (PEG-PC) hydrogel system, and customized it to capture the stiffness and integrin-binding profile of in vivo tumors. We report that the efficacy of a Raf kinase inhibitor, sorafenib, is reduced on stiff, collagen-rich microenvironments, independent of ROCK activity. Instead, sustained activation of JNK mediated this resistance, and combining a JNK inhibitor with sorafenib eliminated stiffness-mediated resistance in triple negative breast cancer cells. Surprisingly, neither ERK nor p38 appears to mediate sorafenib resistance, and instead, either ERK or p38 inhibition rescued sorafenib resistance during JNK inhibition, suggesting negative crosstalk between these signaling pathways on stiff, collagen-rich environments. Overall, we discovered that β1integrin and its downstream effector JNK mediate sorafenib resistance during tumor stiffening. These results also highlight the need for more advanced cell culture platforms, such as our high-throughput PEG-PC system, with which to screen chemotherapeutics

Chemical Vapor Deposition Model of Polysilicon in a Trichlorosilane and Hydrogen System

The traditional polysilicon processes should be refined when addressing the low energy consumption requirement for the production of solar grade silicon. This paper addresses the fluid dynamic conditions required to deposit polysilicon in the traditional Siemens reactor. Analytical solutions for the deposition process are presented, providing information on maximizing the rate between the amount of polysilicon obtained and the energy consumed during the deposition process. The growth rate, deposition efficiency, and power-loss dependence on the gas velocity, the mixture of gas composition, the reactor pressure, and the surface temperature have been analyzed. The analytical solutions have been compared to experimental data and computational solutions presented in the literature. At atmospheric pressure, the molar fraction of hydrogen at the inlet should be adjusted to the range of 0.85–0.90, the gas inlet temperature should be raised within the interval of 673 and 773 K, and the gas velocity should reach the Reynolds number 800. The resultant growth rate will be between 6 and 6.5 _m min−1. Operation above atmospheric pressure is strongly recommended to achieve growth rates of 20 _m min−1 at 6 atm

PEG-Phosphorylcholine Hydrogels As Tunable and Versatile Platforms for Mechanobiology

We report here the synthesis of a new class of hydrogels with an extremely wide range of mechanical properties suitable for cell studies. Mechanobiology has emerged as an important field in bioengineering, in part due to the development of synthetic polymer gels and fibrous protein biomaterials to control and quantify how cells sense and respond to mechanical forces in their microenvironment. To address the problem of limited availability of biomaterials, in terms of both mechanical range and optical clarity, we have prepared hydrogels that combine poly(ethylene glycol) (PEG) and phosphorylcholine (PC) zwitterions. Our goal was to create a hydrogel platform that exceeds the range of Young’s moduli reported for similar hydrogels, while being simple to synthesize and manipulate. The Young’s modulus of these “PEG-PC” hydrogels can be tuned over 4 orders of magnitude, much greater than commonly used hydrogels such as PEG-diacrylate, PEG-dimethacrylate, and polyacrylamide, with smaller average mesh sizes and optical clarity. We prepared PEG-PC hydrogels to study how substrate mechanical properties influence cell morphology, focal adhesion structure, and proliferation across multiple mammalian cell lines, as a proof of concept. These novel PEG-PC biomaterials represent a new and useful class of mechanically tunable hydrogels for mechanobiology

Modularity map of the network of human cell differentiation

Cell differentiation in multicellular organisms is a complex process whose mechanism can be understood by a reductionist approach, in which the individual processes that control the generation of different cell types are identified. Alternatively, a large scale approach in search of different organizational features of the growth stages promises to reveal its modular global structure with the goal of discovering previously unknown relations between cell types. Here we sort and analyze a large set of scattered data to construct the network of human cell differentiation (NHCD) based on cell types (nodes) and differentiation steps (links) from the fertilized egg to a crying baby. We discover a dynamical law of critical branching, which reveals a fractal regularity in the modular organization of the network, and allows us to observe the network at different scales. The emerging picture clearly identifies clusters of cell types following a hierarchical organization, ranging from sub-modules to super-modules of specialized tissues and organs on varying scales. This discovery will allow one to treat the development of a particular cell function in the context of the complex network of human development as a whole. Our results point to an integrated large-scale view of the network of cell types systematically revealing ties between previously unrelated domains in organ functions.Comment: 32 pages, 7 figure