No abstract

A s the drug discovery process evolves and demands more challenging and relevant assays, we have experienced a recent and significant escalation in the number of cell-based high-throughput assays for both small molecule and target identification screens. This has resulted in an increased need for the reproducible production of high quality cells in large quantities. Historically, manual cell culture was the only option available for providing cells in sufficient numbers for small molecule ultra-high-throughput screens (uHTSs), representing a technology gap in our automated cell culture process.Recently, we have applied an automated solution in the form of a novel 10-layer tissue culture flask, the HYPERFlask (Corning, Lowell, MA). This technology, when introduced as an upgrade to the SelecT (an automated cell culture system manufactured by The Automation Partnership, Ltd., Royston, England), provides a new approach to automating production of the high number of cells required for uHTS and consequently a highly desirable alternative to manual cell culture.The HYPERFlask has a surface area of 1720 cm 2 and can yield up to 3 Â 10 8 cells after 3 days in culture. This can be compared to a typical yield of 2 Â 10 7 cells from a T175 flask (with a surface area of 175 cm 2 ), the current standard flask type for automated cell culture on the SelecT. Cells grown in both flask types are of comparable quality, as demonstrated by equivalent cell viability, yield per cm 2 , functional response, and pharmacology.

SYNOPSIS The monthly variation in the incidence of asthma is closely related to temperature. This study, based on the analysis of admissions to children's hospitals throughout Australia, confirms, for a wide range of climates, observations previously reported from Brisbane. The relation is twofold. In the months which normally have a mean temperature below 70°F (21.1°C), asthma increases with increase in temperature. When the temperature exceeds 70°, asthma decreases with increase in temperature, and this is further supported by observations in North Queensland. The incidence of asthma is also related to rainfall. In Perth, where most rain falls in winter, the major incidence of asthma follows in spring. In Brisbane, where most rain falls in summer, the major incidence of asthma follows in autumn. It is suggested that the geographical variation in the incidence of asthma may also be related to temperature and rainfall.

This paper presents an automated, online approach to anomaly detection in high-content screening assays for pharmaceutical research. Online detection of anomalies is attractive because it offers the possibility of immediate corrective action, early termination, and redesign of assays that may require many hours or days to execute. The proposed approach employs assay-specific image processing within an assay-independent framework for distributed control, machine learning, and anomaly reporting. Specifically, we exploit coarse-grained parallelism to distribute image processing over several computing nodes while efficiently aggregating sufficient statistics across nodes. This architecture also allows us to easily handle geographically-distributed data sources. Our results from two applications, adipocyte quantitation and neurite growth estimation, confirm that this online approach to anomaly detection is feasible, efficient, and accurate.

Gene silencing by RNA interference has become a powerful tool to help identify genes that regulate biological processes. However, the complexity of the biology probed and the incomplete validation of the reagents used make it difficult to interpret the results of genome-wide siRNA screens. To address this challenge and maximize the return on the efforts required for validating genomic screen hits, the screening strategy must be designed to increase the robustness of the primary screening hits and include assays that inform on the mechanism of action of the knocked-down transcripts. Here, we describe the implementation of a small interfering RNA (siRNA) screen to identify genes that sensitize the effect of poly-(ADP ribose)-polymerase (PARP) inhibitor on cell survival. In the strategy we designed for the primary screen, two biological activities, apoptosis and cell viability, were measured simultaneously at different time points in the presence and absence of a PARP inhibitor (PARPi). The multiplexed assay allowed us to identify PARPi sensitizers induced by both caspase-dependent and independent mechanisms. The multiplexed screening strategy yielded robust primary hits with significant enrichment for DNA repair genes, which were further validated using relevant high-content imaging assays and confirmation of transcript knockdown by real-time PCR (rtPCR).