OPTIMIZING A SHORT-END ELECTROPHORETICALLY MEDIATED MICRO-ANALYSIS (EMMA) ASSAY FOR CREATININE

In this work electrophoretically mediated micro-analysis (EMMA) is used in conjunction with short end injection to improve the in-capillary Jaffé assay for creatinine. Key advances over prior work include (i) using simulation to ensure intimate overlap of reagent plugs, (ii) using OH- to drive the reaction, (iii) using short-end injection to minimize analysis time and in-line product degradation. The potential-driven overlapping time with the EMMA approach, as well as the borate buffer background electrolyte (BGE) concentration and pH are optimized with the short end approach. The best conditions for short-end analyses would not have been predicted by the prior long end work, owing to a complex interplay of separation time and product degradation rates. Raw peak areas and flow-adjusted peak areas for the Jaffé reaction product (at 505 nm) are used to assess the sensitivity of the short-end EMMA approach. Optimal overlap conditions depend heavily on local conductivity differences within the reagent zone(s), as these differences cause dramatic voltage field differences, which effect reagent overlap dynamics. Simul 5.0, a dynamic simulation program for capillary electrophoresis (CE) systems, is used to understand the ionic boundaries and profiles that give rise to the experimentally obtained data for EMMA analysis. Overall, fast migration of hydroxide ions from the picrate zone makes difficult reagent overlap. In addition, the challenges associated with the simultaneous overlapping of three reagent zones are considered, and experimental results validate the predictions made by the simulation. With one set of “optimized” conditions including OH- (253 mM) as the third reagent zone the response was linear with creatinine concentration (R2 = 0.998) and reproducible over the clinically relevant range (0.08 to 0.1 mM) of standard creatinine concentrations. An LOD (S/N = 3) of 0.02 mM and LOQ (S/N=10) of 0.08 mM were determined. A significant improvement (43%) in assay sensitivity was obtained compared to prior work that considered only two reagents in the overlap.

The Experience of Micro and Macro Events: A Life-Span Analysis

This study investigated how individuals retrospectively construe their lives in terms of major life events. Ninety-nine participants sorted a set of personal and historical events in terms of perceived importance for their lives. Analyses of variance with repeated measures and rank comparisons were computed. Overall findings revealed no cohort differences with regard to the perception of life events. However, within-cohort differences were found, indicating that more life events were recalled from the young adult years. Those experiences were also perceived as having been more important in the participants' lives than events from other age segments. With regard to historical events, war-related experiences were among the highest ranked. Analyses of variance revealed intracohort differences but not intercohort differences, indicating higher scores for the time between 1930 and 1948 relative to other historical periods.

Investigation of the Design of a Bistable Micro-Chemical-Mechanical Device Utilizing Lateral Buckling

The Gracias Laboratory at Johns Hopkins University has developed microgrippers which utilize chemically-actuated joints to be used in micro-surgery. These grippers, however, take up to thirty minutes to close fully when activated biochemicals in the human body. This is very problematic and could limit the use of the devices in surgery. It is the goal of this research to develop a gripper that uses theGracias Laboratory's existing joints in conjunction with mechanical components to decrease the closing time. The purpose of including the mechanical components is to induce a state of instability at which time a small perturbation would cause the joint to close fully.The main concept of the research was to use the lateral buckling of a triangular gripper geometry and use a toggle mechanism to decrease the closure time of the device. This would create a snap-action device mimicking the quick closure of a Venus flytrap. All developed geometries were tested using finite element analysis to determine ifloading conditions produced the desired buckled shape. This research examines lateral buckling on the micro-scale and the possibility ofusing this phenomenon in a micro-gripper. Although a final geometry with the required deformed shaped was not found, this document contains suggestions for future geometries that may produce the correct deformed shape. It was determined through this work that in order to obtain the desired deformed shape, polymeric sections need to be added to the geometry. This simplifies the analysis and allows the triangular structure to buckle in the appropriate way due to the added joints. Future work for this project will be completed by undergraduate students at Bucknell University. Fabrication and testing of devices will be done at Johns Hopkins University in the Gracias Laboratory.

Using Micro-Gravity Techniques to Map Alluvium Thickness and Pleistocene Location of the West Branch of the Susquehanna River Near Muncy, Pennsylvania

Laurentide glaciation during the early Pleistocene (~970 ka) dammed the southeast-flowing West Branch of the Susquehanna River (WBSR), scouring bedrock and creating 100-km-long glacial Lake Lesley near the Great Bend at Muncy, Pennsylvania (Ramage et al., 1998). Local drill logs and well data indicate that subsequent paleo-outwash floods and modern fluvial processes have deposited as much as 30 meters of alluvium in this area, but little is known about the valley fill architecture and the bedrock-alluvium interface. By gaining a greater understanding of the bedrock-alluvium interface the project will not only supplement existing depth to bedrock information, but also provide information pertinent to the evolution of the Muncy Valley landscape. This project determined if variations in the thickness of the valley fill were detectable using micro-gravity techniques to map the bedrock-alluvium interface. The gravity method was deemed appropriate due to scale of the study area (~30 km2), ease of operation by a single person, and the available geophysical equipment. A LaCoste and Romberg Gravitron unit was used to collect gravitational field readings at 49 locations over 5 transects across the Muncy Creek and Susquehanna River valleys (approximately 30 km2), with at least two gravity base stations per transect. Precise latitude, longitude and ground surface elevation at each location were measured using an OPUS corrected Trimble RTK-GPS unit. Base stations were chosen based on ease of access due to the necessity of repeat measurements. Gravity measurement locations were selected and marked to provide easy access and repeat measurements. The gravimeter was returned to a base station within every two hours and a looping procedure was used to determine drift and maximize confidence in the gravity measurements. A two-minute calibration reading at each station was used to minimize any tares in the data. The Gravitron digitally recorded finite impulse response filtered gravity measurements every 20 seconds at each station. A measurement period of 15 minutes was used for each base station occupation and a minimum of 5 minutes at all other locations. Longer or multiple measurements were utilized at some sites if drift or other externalities (i.e. train or truck traffic) were effecting readings. Average, median, standard deviation and 95% confidence interval were calculated for each station. Tidal, drift, latitude, free-air, Bouguer and terrain corrections were then applied. The results show that the gravitational field decreases as alluvium thickness increases across the axes of the Susquehanna River and Muncy Creek valleys. However, the location of the gravity low does not correspond with the present-day location of the West Branch of the Susquehanna River (WBSR), suggesting that the WBSR may have been constrained along Bald Eagle Mountain by a glacial lobe originating from the Muncy Creek Valley to the northeast. Using a 3-D inversion model, the topography of the bedrock-alluvium interface was determined over the extent of the study area using a density contrast of -0.8 g/cm3. Our results are consistent with the bedrock geometry of the area, and provide a low-cost, non-invasive and efficient method for exploring the subsurface and for supplementing existing well data.