Gradient microfluidics enables rapid bacterial growth inhibition testing

Bacterial growth inhibition tests have become a standard measure of the adverse effects of inhibitors for a wide range of applications, such as toxicity testing in the medical and environmental sciences. However, conventional well-plate formats for these tests are laborious and provide limited information (often being restricted to an end-point assay). In this study, we have developed a microfluidic system that enables fast quantification of the effect of an inhibitor on bacteria growth and survival, within a single experiment. This format offers a unique combination of advantages, including long-term continuous flow culture, generation of concentration gradients, and single cell morphology tracking. Using Escherichia coli and the inhibitor amoxicillin as one model system, we show excellent agreement between an on-chip single cell-based assay and conventional methods to obtain quantitative measures of antibiotic inhibition (for example, minimum inhibition concentration). Furthermore, we show that our methods can provide additional information, over and above that of the standard well-plate assay, including kinetic information on growth inhibition and measurements of bacterial morphological dynamics over a wide range of inhibitor concentrations. Finally, using a second model system, we show that this chip-based systems does not require the bacteria to be labeled and is well suited for the study of naturally occurring species. We illustrate this using Nitrosomonas europaea, an environmentally important bacteria, and show that the chip system can lead to a significant reduction in the period required for growth and inhibition measurements (<4 days, compared to weeks in a culture flask)

Single Cell Deposition and Patterning with a Robotic System

Integrating single-cell manipulation techniques in traditional and emerging biological culture systems is challenging. Microfabricated devices for single cell studies in particular often require cells to be spatially positioned at specific culture sites on the device surface. This paper presents a robotic micromanipulation system for pick-and-place positioning of single cells. By integrating computer vision and motion control algorithms, the system visually tracks a cell in real time and controls multiple positioning devices simultaneously to accurately pick up a single cell, transfer it to a desired substrate, and deposit it at a specified location. A traditional glass micropipette is used, and whole- and partial-cell aspiration techniques are investigated to manipulate single cells. Partially aspirating cells resulted in an operation speed of 15 seconds per cell and a 95% success rate. In contrast, the whole-cell aspiration method required 30 seconds per cell and achieved a success rate of 80%. The broad applicability of this robotic manipulation technique is demonstrated using multiple cell types on traditional substrates and on open-top microfabricated devices, without requiring modifications to device designs. Furthermore, we used this serial deposition process in conjunction with an established parallel cell manipulation technique to improve the efficiency of single cell capture from ∼80% to 100%. Using a robotic micromanipulation system to position single cells on a substrate is demonstrated as an effective stand-alone or bolstering technology for single-cell studies, eliminating some of the drawbacks associated with standard single-cell handling and manipulation techniques

Enhancing apoptosis in TRAIL-resistant cancer cells using fundamental response rules

The tumor necrosis factor related apoptosis-inducing ligand (TRAIL) induces apoptosis in malignant cells, while leaving other cells mostly unharmed. However, several carcinomas remain resistant to TRAIL. To investigate the resistance mechanisms in TRAIL-stimulated human fibrosarcoma (HT1080) cells, we developed a computational model to analyze the temporal activation profiles of cell survival (IκB, JNK, p38) and apoptotic (caspase-8 and -3) molecules in wildtype and several (FADD, RIP1, TRAF2 and caspase-8) knock-down conditions. Based on perturbation-response approach utilizing the law of information (signaling flux) conservation, we derived response rules for population-level average cell response. From this approach, i) a FADD-independent pathway to activate p38 and JNK, ii) a crosstalk between RIP1 and p38, and iii) a crosstalk between p62 and JNK are predicted. Notably, subsequent simulations suggest that targeting a novel molecule at p62/sequestosome-1 junction will optimize apoptosis through signaling flux redistribution. This study offers a valuable prospective to sensitive TRAIL-based therapy

The impact of sexual harassment on job satisfaction, turnover intentions, and absenteeism: findings from Pakistan compared to the United States

The purpose of this study was to compare and contrast how differences in perceptions of sexual harassment impact productive work environments for employees in Pakistan as compared to the US; in particular, how it affects job satisfaction, turnover, and/or absenteeism. This study analyzed employee responses in Pakistan (n = 146) and the United States (n = 102, 76) using questionnaire data. Significant results indicated that employees who were sexually harassed reported (a) a decrease in job satisfaction (b) greater turnover intentions and (c) a higher rate of absenteeism. Cross-cultural comparisons indicated that (a) Pakistani employees who were sexually harassed had greater job dissatisfaction and higher overall absenteeism than did their US counterparts and (b) Pakistani women were more likely to use indirect strategies to manage sexual harassment than were US targets

Multilayer buffer for high-temperature superconductor devises on MgO

A multilayer thin film epitaxial passivation of single crystal MgO substrates was developed. YBa2Cu3O7-x films on the buffered MgO substrates demonstrate pure c-axis orientation, absence of in-plane disoriented grains, transition temperature T-c > 91 K, and critical current density J(c) similar to 5 MA/cm(2) at 77.4 K and were deposited in thicknesses of up to several micrometers without cracks. High-temperature superconductor multilayer flux transformers of 2 mu m thickness on the buffered MgO substrates demonstrated improved insulation between the superconducting layers and an increased dynamic range compared to flux transformers on SrTiO3 substrates. ((c))2006 American Institute of Physics

Properties of YBa2Cu3O7 thin films deposited on substrates and bicrystals with vicinal offcut and realization of high IcRn Junctions

The surface morphology, microstructure and transport properties of epitaxial YBa2Cu3O7 and PrBa2Cu3O7 thin films and heterostructures deposited on slightly vicinal substrates of SrTiO3 by high pressure oxygen sputtering were studied. The vicinal angles of the substrates and bicrystals were less then 13 degrees. Depending on the tilt angle of the substrate a transition from spiral or island to step-flow growth leading to an improvement of the surface roughness was observed. Atomic force and transmission electron microscopy were used for these investigations. Furthermore, electrical and structural properties of YBa2Cu3O7 thin films on vicinal offcut SrTiO3 bicrystals with different grain boundary types were studied. This included junctions with a 2x 12 degrees tilt or twist of the YBa2Cu3O7 c-axis across the grain boundary. In comparison to conventional [001]-tilt grain boundaries bicrystal Josephson junctions [100]-tilt grain boundaries showed high I-c R-n -products of up to 1.2 meV at 77 K and up to 8 meV at 4.2 K. IV-curve instabilities, probably of magnetic origin due to flux flow in the electrodes, often could he observed for junctions biased with high current densities