Establishing the C. elegans uterine seam cell (utse) as a novel model for studying cell behavior

The molecular inputs necessary for cell behavior are vital to our understanding of development and disease. Proper cell behavior is necessary for processes ranging from creating one’s face (neural crest migration) to spreading cancer from one tissue to another (invasive metastatic cancers). Identifying the genes and tissues involved in cell behavior not only increases our understanding of biology but also has the potential to create targeted therapies in diseases hallmarked by aberrant cell behavior.

A well-characterized model system is key to determining the molecular and spatial inputs necessary for cell behavior. In this work I present the C. elegans uterine seam cell (utse) as an ideal model for studying cell outgrowth and shape change. The utse is an H-shaped cell within the hermaphrodite uterus that functions in attaching the uterus to the body wall. Over L4 larval stage, the utse grows bidirectionally along the anterior-posterior axis, changing from an ellipsoidal shape to an elongated H-shape. Spatially, the utse requires the presence of the uterine toroid cells, sex muscles, and the anchor cell nucleus in order to properly grow outward. Several gene families are involved in utse development, including Trio, Nav, Rab GTPases, Arp2/3, as well as 54 other genes found from a candidate RNAi screen. The utse can be used as a model system for studying metastatic cancer. Meprin proteases are involved in promoting invasiveness of metastatic cancers and the meprin-likw genes nas-21, nas-22, and toh-1 act similarly within the utse. Studying nas-21 activity has also led to the discovery of novel upstream inhibitors and activators as well as targets of nas-21, some of which have been characterized to affect meprin activity. This illustrates that the utse can be used as an in vivo model for learning more about meprins, as well as various other proteins involved in metastasis.

An investigation of various types of arc welded joints

In the last few years there has been much discussion as to the relative merits or arc welded joints as compared to riveted joints. There is an old precedent which must be removed before arc welding will come into its own. The purpose of this paper is to help establish confidence in this process of fabrication with special regard to strength and reliability.

Developing and characterizing bulk metallic glasses for extreme applications

Metallic glasses have typically been treated as a “one size fits all” type of material. Every alloy is considered to have high strength, high hardness, large elastic limits, corrosion resistance, etc. However, similar to traditional crystalline materials, properties are strongly dependent upon the constituent elements, how it was processed, and the conditions under which it will be used. An important distinction which can be made is between metallic glasses and their composites. Charpy impact toughness measurements are performed to determine the effect processing and microstructure have on bulk metallic glass matrix composites (BMGMCs). Samples are suction cast, machined from commercial plates, and semi-solidly forged (SSF). The SSF specimens have been found to have the highest impact toughness due to the coarsening of the dendrites, which occurs during the semi-solid processing stages. Ductile to brittle transition (DTBT) temperatures are measured for a BMGMC. While at room temperature the BMGMC is highly toughened compared to a fully glassy alloy, it undergoes a DTBT by 250 K. At this point, its impact toughness mirrors that of the constituent glassy matrix. In the following chapter, BMGMCs are shown to have the capability of being capacitively welded to form single, monolithic structures. Shear measurements are performed across welded samples, and, at sufficient weld energies, are found to retain the strength of the parent alloy. Cross-sections are inspected via SEM and no visible crystallization of the matrix occurs.

Next, metallic glasses and BMGMCs are formed into sheets and eggbox structures are tested in hypervelocity impacts. Metallic glasses are ideal candidates for protection against micrometeorite orbital debris due to their high hardness and relatively low density. A flat single layer, flat BMG is compared to a BMGMC eggbox and the latter creates a more diffuse projectile cloud after penetration. A three tiered eggbox structure is also tested by firing a 3.17 mm aluminum sphere at 2.7 km/s at it. The projectile penetrates the first two layers, but is successfully contained by the third.

A large series of metallic glass alloys are created and their wear loss is measured in a pin on disk test. Wear is found to vary dramatically among different metallic glasses, with some considerably outperforming the current state-of-the-art crystalline material (most notably Cu₄₃Zr₄₃Al₇Be₇). Others, on the other hand, suffered extensive wear loss. Commercially available Vitreloy 1 lost nearly three times as much mass in wear as alloy prepared in a laboratory setting. No conclusive correlations can be found between any set of mechanical properties (hardness, density, elastic, bulk, or shear modulus, Poisson’s ratio, frictional force, and run in time) and wear loss. Heat treatments are performed on Vitreloy 1 and Cu₄₃Zr₄₃Al₇Be₇. Anneals near the glass transition temperature are found to increase hardness slightly, but decrease wear loss significantly. Crystallization of both alloys leads to dramatic increases in wear resistance. Finally, wear tests under vacuum are performed on the two alloys above. Vitreloy 1 experiences a dramatic decrease in wear loss, while Cu₄₃Zr₄₃Al₇Be₇ has a moderate increase. Meanwhile, gears are fabricated through three techniques: electrical discharge machining of 1 cm by 3 mm cylinders, semisolid forging, and copper mold suction casting. Initial testing finds the pin on disk test to be an accurate predictor of wear performance in gears.

The final chapter explores an exciting technique in the field of additive manufacturing. Laser engineered net shaping (LENS) is a method whereby small amounts of metallic powders are melted by a laser such that shapes and designs can be built layer by layer into a final part. The technique is extended to mixing different powders during melting, so that compositional gradients can be created across a manufactured part. Two compositional gradients are fabricated and characterized. Ti 6Al¬ 4V to pure vanadium was chosen for its combination of high strength and light weight on one end, and high melting point on the other. It was inspected by cross-sectional x-ray diffraction, and only the anticipated phases were present. 304L stainless steel to Invar 36 was created in both pillar and as a radial gradient. It combines strength and weldability along with a zero coefficient of thermal expansion material. Only the austenite phase is found to be present via x-ray diffraction. Coefficient of thermal expansion is measured for four compositions, and it is found to be tunable depending on composition.

Three-dimensional nonlinear inelastic analysis of steel moment-frame buildings damaged by earthquake excitations

The Northridge earthquake of January 17, 1994, highlighted the two previously known problems of premature fracturing of connections and the damaging capabilities of near-source ground motion pulses. Large ground motions had not been experienced in a city with tall steel moment-frame buildings before. Some steel buildings exhibited fracture of welded connections or other types of structural degradation.

A sophisticated three-dimensional nonlinear inelastic program is developed that can accurately model many nonlinear properties commonly ignored or approximated in other programs. The program can assess and predict severely inelastic response of steel buildings due to strong ground motions, including collapse.

Three-dimensional fiber and segment discretization of elements is presented in this work. This element and its two-dimensional counterpart are capable of modeling various geometric and material nonlinearities such as moment amplification, spread of plasticity and connection fracture. In addition to introducing a three-dimensional element discretization, this work presents three-dimensional constraints that limit the number of equations required to solve various three-dimensional problems consisting of intersecting planar frames.

Two buildings damaged in the Northridge earthquake are investigated to verify the ability of the program to match the level of response and the extent and location of damage measured. The program is used to predict response of larger near-source ground motions using the properties determined from the matched response.

A third building is studied to assess three-dimensional effects on a realistic irregular building in the inelastic range of response considering earthquake directivity. Damage levels are observed to be significantly affected by directivity and torsional response.

Several strong recorded ground motions clearly exceed code-based levels. Properly designed buildings can have drifts exceeding code specified levels due to these ground motions. The strongest ground motions caused collapse if fracture was included in the model. Near-source ground displacement pulses can cause columns to yield prior to weaker-designed beams. Damage in tall buildings correlates better with peak-to-peak displacements than with peak-to-peak accelerations.

Dynamic response of tall buildings shows that higher mode response can cause more damage than first mode response. Leaking of energy between modes in conjunction with damage can cause torsional behavior that is not anticipated.

Various response parameters are used for all three buildings to determine what correlations can be made for inelastic building response. Damage levels can be dramatically different based on the inelastic model used. Damage does not correlate well with several common response parameters.

Realistic modeling of material properties and structural behavior is of great value for understanding the performance of tall buildings due to earthquake excitations.

The Flocculation of E. coli with Polyethyleneimine

A comprehensive study was made of the flocculation of dispersed E. coli bacterial cells by the cationic polymer polyethyleneimine (PEI). The three objectives of this study were to determine the primary mechanism involved in the flocculation of a colloid with an oppositely charged polymer, to determine quantitative correlations between four commonly-used measurements of the extent of flocculation, and to record the effect of varying selected system parameters on the degree of flocculation. The quantitative relationships derived for the four measurements of the extent of flocculation should be of direct assistance to the sanitary engineer in evaluating the effectiveness of specific coagulation processes.

A review of prior statistical mechanical treatments of absorbed polymer configuration revealed that at low degrees of surface site coverage, an oppositely- charged polymer molecule is strongly adsorbed to the colloidal surface, with only short loops or end sequences extending into the solution phase. Even for high molecular weight PEI species, these extensions from the surface are theorized to be less than 50 Å in length. Although the radii of gyration of the five PEI species investigated were found to be large enough to form interparticle bridges, the low surface site coverage at optimum flocculation doses indicates that the predominant mechanism of flocculation is adsorption coagulation.

The effectiveness of the high-molecular weight PEI species 1n producing rapid flocculation at small doses is attributed to the formation of a charge mosaic on the oppositely-charged E. coli surfaces. The large adsorbed PEI molecules not only neutralize the surface charge at the adsorption sites, but also cause charge reversal with excess cationic segments. The alignment of these positive surface patches with negative patches on approaching cells results in strong electrostatic attraction in addition to a reduction of the double-layer interaction energies. The comparative ineffectiveness of low-molecular weight PEI species in producing E. coli flocculation is caused by the size of the individual molecules, which is insufficient to both neutralize and reverse the negative E.coli surface charge. Consequently, coagulation produced by low molecular weight species is attributed solely to the reduction of double-layer interaction energies via adsorption.

Electrophoretic mobility experiments supported the above conclusions, since only the high-molecular weight species were able to reverse the mobility of the E. coli cells. In addition, electron microscope examination of the seam of agglutination between E. coli cells flocculation by PEI revealed tightly- bound cells, with intercellular separation distances of less than 100-200 Å in most instances. This intercellular separation is partially due to cell shrinkage in preparation of the electron micrographs.

The extent of flocculation was measured as a function of PEl molecular weight, PEl dose, and the intensity of reactor chamber mixing. Neither the intensity of mixing, within the common treatment practice limits, nor the time of mixing for up to four hours appeared to play any significant role in either the size or number of E.coli aggregates formed. The extent of flocculation was highly molecular weight dependent: the high-molecular-weight PEl species produce the larger aggregates, the greater turbidity reductions, and the higher filtration flow rates. The PEl dose required for optimum flocculation decreased as the species molecular weight increased. At large doses of high-molecular-weight species, redispersion of the macroflocs occurred, caused by excess adsorption of cationic molecules. The excess adsorption reversed the surface charge on the E.coli cells, as recorded by electrophoretic mobility measurements.

Successful quantitative comparisons were made between changes in suspension turbidity with flocculation and corresponding changes in aggregate size distribution. E. coli aggregates were treated as coalesced spheres, with Mie scattering coefficients determined for spheres in the anomalous diffraction regime. Good quantitative comparisons were also found to exist between the reduction in refiltration time and the reduction of the total colloid surface area caused by flocculation. As with turbidity measurements, a coalesced sphere model was used since the equivalent spherical volume is the only information available from the Coulter particle counter. However, the coalesced sphere model was not applicable to electrophoretic mobility measurements. The aggregates produced at each PEl dose moved at approximately the same vlocity, almost independently of particle size.

PEl was found to be an effective flocculant of E. coli cells at weight ratios of 1 mg PEl: 100 mg E. coli. While PEl itself is toxic to E.coli at these levels, similar cationic polymers could be effectively applied to water and wastewater treatment facilities to enhance sedimentation and filtration characteristics.

A retrofitting framework for pre-Northridge steel moment-frame buildings

In the 1994 M_w 6.7 Northridge and 1995 M_w 6.9 Kobe earthquakes, steel moment-frame buildings were exposed to an unexpected flaw. The commonly utilized welded unreinforced flange, bolted web connections were observed to experience brittle fractures in a number of buildings, even at low levels of seismic demand. A majority of these buildings have not been retrofitted and may be susceptible to structural collapse in a major earthquake.

This dissertation presents a case study of retrofitting a 20-story pre-Northridge steel moment-frame building. Twelve retrofit schemes are developed that present some range in degree of intervention. Three retrofitting techniques are considered: upgrading the brittle beam-to-column moment resisting connections, and implementing either conventional or buckling-restrained brace elements within the existing moment-frame bays. The retrofit schemes include some that are designed to the basic safety objective of ASCE-41 Seismic Rehabilitation of Existing Buildings.

Detailed finite element models of the base line building and the retrofit schemes are constructed. The models include considerations of brittle beam-to-column moment resisting connection fractures, column splice fractures, column baseplate fractures, accidental contributions from ``simple'' non-moment resisting beam-to-column connections to the lateral force-resisting system, and composite actions of beams with the overlying floor system. In addition, foundation interaction is included through nonlinear translational springs underneath basement columns.

To investigate the effectiveness of the retrofit schemes, the building models are analyzed under ground motions from three large magnitude simulated earthquakes that cause intense shaking in the greater Los Angeles metropolitan area, and under recorded ground motions from actual earthquakes. It is found that retrofit schemes that convert the existing moment-frames into braced-frames by implementing either conventional or buckling-restrained braces are effective in limiting structural damage and mitigating structural collapse. In the three simulated earthquakes, a 20% chance of simulated collapse is realized at PGV of around 0.6 m/s for the base line model, but at PGV of around 1.8 m/s for some of the retrofit schemes. However, conventional braces are observed to deteriorate rapidly. Hence, if a braced-frame that employs conventional braces survives a large earthquake, it is questionable how much service the braces provide in potential aftershocks.