Automated High Throughput Screening of Fluorescent Imaging Plate Reader (FLIPR) Assays Using Assay Platform™ Integrated with Activity Base for ‘on the fly’ Compound Reconfirmation

Advances in the field of automation have meant hitherto complex manual cell-based assays can now be automated. These improvements have brought significant enhancements in throughput, data fidelity and consistency, and allowed a reallocation of constrained resources.Building upon these improvements, we have linked our automated cell-based screening system, Assay Platform™, to Activity Base (IDBS), a software package designed to automate the analysis of HTS data. Customisation of this package has resulted in software that can identify ‘active’ compounds and re-pick them ‘on the fly’ from the original compound plates for triplicate re-testing without operator intervention.Based on an operator initially defining ‘normal’ parameters for assay activity in Activity Base, combined with an automated quality control software module that checks data fidelity, wells containing ‘active’ compounds can be re-picked and re-tested at the end of an automated screening run. Automating cell-based assays has significantly improved productivity, and, with the synergism of Activity Base, has given us greater power to complete each screening run and report ‘active’ compounds to Chemistry more rapidly. This article presents our approach to the automation of cell-based Fluorescent Imaging Plate Reader (FLIPR) screening together with automated active re-test confirmation using Activity Base.     

Silver Bullet Talks with Ed Amoroso

Gary McGraw interviews Ed Amoroso, AT&T's chief information security officer. Their conversation ranged widely, from system design to privacy.     

Top-down proteomic analysis by MALDI-TOF profiling: Concentration-independent biomarkers

Although numerous protein biomarkers have been correlated with advanced disease states, no new clinical assays have been developed. Goals often anticipate disease-specific protein changes that exceed values among healthy individuals, a property common to acute phase reactants. This review considers somewhat different approaches. It focuses on intact protein isoform ratios that present a biomarker without change in the total concentration of the protein. These will seldom be detected by peptide level analysis or by most antibody-based assays. For example, application of an inexpensive method to large sample groups resulted in observation of several polymorphisms, including the first structural polymorphism of apolipoprotein C1. Isoform distribution of this protein was altered and was eventually linked to increased obesity. Numerous other protein isoforms included C- and N-terminal proteolysis, changes of glycoisoform ratios and certain types of sulfhydryl oxidation. While many of these gave excellent statistical correlation with advanced disease, clinical utility was not apparent. More important may be that protein isoform ratios were very stable in each individual. Diagnosis by longitudinal analysis of the same individual might increase sensitivity of protein biomarkers by 20-fold or more. Protein changes that exceed the range of values found among healthy individuals may be uncommon.     

An evaluation of backpack harness systems in non-neutral torso postures

Much of the research on backpack design has been focused on spinal loading/biomechanics while the wearer is in a neutral/upright trunk posture, such as those employed by outdoor enthusiasts and schoolchildren. This research has led to some important harness design improvements that reduce trunk muscle exertions, fatigue and improve overall comfort. There are number of occupations, however, wherein workers wear back-mounted packs/devices (e.g. air tanks) while working in non-neutral trunk postures. The objective of the current study was to evaluate the effects of these non-neutral postures on biomechanical loading and then reconsider the backpack system design recommendations. Fifteen participants were asked to support a 18.2 kg load on their back while assuming static forward flexed postures of the torso (15 degrees , 30 degrees , 45 degrees , and 60 degrees of sagittal bend). The mass on the back was attached to the participant through two different harness mechanisms: a basic harness design (as seen on college student backpacks) and a more advanced design containing lateral stiffness rods and a weight-bearing hip belt (as seen on backpacks for hikers). While performing these static, posture maintenance tasks, the activation levels of the bilateral trapezius, erector spinae, and rectus abdominis were collected. Participants also provided subjective ratings of comfort. The results showed that there was a significant interaction between harness type and forward flexion angle for the trapezius and the erector spinae muscles. The normalized EMG for the trapezius muscles showed a 14% and 11% reduction in muscle activity at 15 degrees and 30 degrees , respectively, with the advanced design but these positive effects of the advanced design were not found at the greater flexion angles. Likewise the erector spinae muscles showed a 24% and 14% reduction in muscle activity at 15 degrees and 30 degrees , respectively, with the advanced design harness but these effects of the advanced design were not found at the greater forward flexion angles. The level of forward flexion angle affected the rectus abdominis muscle activity, but neither the harness type main effect nor the interaction of harness type and forward flexion angle was significant. The subjective survey results agreed with the EMG results and showed the advanced design harness was generally more comfortable with respect to the shoulder and low back areas. Collectively, the subjective and objective results show a significant improvement with the advanced harness system but also note an interesting interaction with degree of sagittal flexion, indicating a diminished effectiveness of the design improvements at forward flexed postures. Design criteria for harness systems in these forward flexed postures are discussed.     

Vortex dynamics at the superfluid λ-transition

A vortex-ring theory of the superfluid⁴He -transition is extended to include the dynamics of the transition. The response of the vortices to an oscillating superflow is found by solving the Fokker-Planck equation. This allows a calculation of the superfluid relaxation time, which is in agreement with Landau-Khalatnikov theory and with dynamic scaling. At high frequencies the transition becomes broadened, with both the superfluid density and the dissipation remaining finite at and above T. Comparison is made to earlier theories that use high-temperature expansions and renormalization-group expansions. Applications to other subjects such as mutual friction, high-T_c superconductors, and rapidly quenched systems are briefly discussed.     

Ceramic Powder Synthesis by Spray Pyrolysis

A variety of spray pyrolysis (SP) techniques have been developed to directly produce ceramic powders from solutions. This paper reviews the current status of these processes in terms of the process parameters that enable the formation of particles with controlled morphology and composition. A model incorporating solute diffusion in the droplet and solvent evaporation from the droplet surface is presented to establish the critical parameters leading to solid particle formation. The model illustrates that solid particles can be obtained if solutes with high solubility and a large difference between the critical supersaturation and equilibrium concentration are used and if the process is designed to avoid solvent boiling. It is demonstrated that mixed metal oxide, non-oxide, and composite particles that are solid, hollow, porous, or fibrous can be produced by modifying the precursor characteristics, solution properties, and process parameters. The physical and chemical flexibility of SP processes offers numerous opportunities for the controlled synthesis of advanced ceramic powders and films. However, production rates are limited by the need to produce < 5-μm-diameter droplets and to avoid subsequent droplet coagulation. Developments in process controls, atomization, and system design are required for wider commercialization of SP-type processes.     

Recent change in summer chlorophyll a dynamics of southeastern Lake Michigan

Six offshore stations in southeastern Lake Michigan were sampled during a pre quagga mussel Dreissena rostriformis bugensis period (1995–2000) and a post quagga mussel period (2007–2011). Chlorophyll a fluorescence profiles were used to characterize chlorophyll a concentrations during early (June–July) and late (August–September) summer stratification. During the early summer period the average whole water column chlorophyll a, the deep chlorophyll maximum, and the size of deep chlorophyll layer decreased 50%, 55%, and 92%, respectively, between 1995–2000 and 2007–2011. By contrast, in late summer there were no changes in these metrics between periods. Surface mixed layer chlorophyll a in early and late summer did not differ between time periods. On the other hand, chlorophyll a in the near bottom zone (bottom 20 m) declined 63% and 54% between 1995–2000 and 2007–2011 in early and late summer respectively. Changes in total phosphorus between 1995–2000 and 2007–2011 were less dramatic, with declines of 22–27% in early summer and 11–30% in late summer. Changes in the chlorophyll a conditions were attributed to dreissenid mussels which reduced material available from the spring bloom and disrupted the horizontal transport of nutrients to the offshore. Although light availability increased (i.e., increased secchi depths), reduced nutrient availability and spring diatom abundance resulted in a much smaller deep chlorophyll layer in 2007–2011.     

A computational study on the quadratic knapsack problem with multiple constraints

The quadratic knapsack problem (QKP) has been the subject of considerable research in recent years. Despite notable advances in special purpose solution methodologies for QKP, this problem class remains very difficult to solve. With the exception of special cases, the state-of-the-art is limited to addressing problems of a few hundred variables and a single knapsack constraint.In this paper we provide a comparison of quadratic and linear representations of QKP based on test problems with multiple knapsack constraints and up to eight hundred variables. For the linear representations, three standard linearizations are investigated. Both the quadratic and linear models are solved by standard branch-and-cut optimizers available via CPLEX. Our results show that the linear models perform well on small problem instances but for larger problems the quadratic model outperforms the linear models tested both in terms of solution quality and solution time by a wide margin. Moreover, our results demonstrate that QKP instances larger than those previously addressed in the literature as well as instances with multiple constraints can be successfully and efficiently solved by branch and cut methodologies.     

Musculoskeletal disorder risk during automotive assembly: current vs. seated

Musculoskeletal disorder risk was assessed during automotive assembly processes. The risk associated with current assembly processes was compared to using a cantilever chair intervention. Spine loads and normalized shoulder muscle activity were evaluated during assembly in eight regions of the vehicle. Eight interior cabin regions of the vehicle were classified by reach distance, height from vehicle floor and front to back. The cantilever chair intervention tool was most effective in the far reach regions regardless of the height. In the front far reach regions both spine loads and normalized shoulder muscle activity levels were reduced. In the middle and close reach regions spine loads were reduced, however, shoulder muscle activity was not, thus an additional intervention would be necessary to reduce shoulder risk. In the back far reach region, spine loads were not significantly different between the current and cantilever chair conditions. Thus, the effectiveness of the cantilever chair was dependent on the region of the vehicle.