A Barcoding Strategy Enabling Higher-Throughput Library Screening by Microscopy.

Dramatic progress has been made in the design and build phases of the design-build-test cycle for engineering cells. However, the test phase usually limits throughput, as many outputs of interest are not amenable to rapid analytical measurements. For example, phenotypes such as motility, morphology, and subcellular localization can be readily measured by microscopy, but analysis of these phenotypes is notoriously slow. To increase throughput, we developed microscopy-readable barcodes (MiCodes) composed of fluorescent proteins targeted to discernible organelles. In this system, a unique barcode can be genetically linked to each library member, making possible the parallel analysis of phenotypes of interest via microscopy. As a first demonstration, we MiCoded a set of synthetic coiled-coil leucine zipper proteins to allow an 8 × 8 matrix to be tested for specific interactions in micrographs consisting of mixed populations of cells. A novel microscopy-readable two-hybrid fluorescence localization assay for probing candidate interactions in the cytosol was also developed using a bait protein targeted to the peroxisome and a prey protein tagged with a fluorescent protein. This work introduces a generalizable, scalable platform for making microscopy amenable to higher-throughput library screening experiments, thereby coupling the power of imaging with the utility of combinatorial search paradigms.

Pulmonary embolism in the pediatric emergency department: a case demonstrating the application of point-of-care cardiac ultrasound in a pediatric patient with pulmonary embolism.

Although pulmonary embolism in children is rare, it is important for the pediatric emergency medicine provider to be aware of its presentation and emergent management. We present a case of bilateral pulmonary embolisms in an adolescent patient to illustrate the benefits from the timely diagnosis of right ventricular dysfunction by point-of-care echocardiography performed by emergency medicine physicians. Ultrasonographic techniques and the emergent management of pulmonary embolism are reviewed.

Psychological resilience and dysfunction among hospitalized survivors of the SARS epidemic in Hong Kong: a latent class approach.

OBJECTIVE: To examine trajectories of psychological functioning using latent class analysis on a sample of hospitalized survivors of the 2003 severe acute respiratory syndrome (SARS) epidemic in Hong Kong. DESIGN: A longitudinal study of 997 survivors, recruited from among 1,331 individuals hospitalized for SARS, were interviewed at 6, 12, and 18 months after hospitalization. MAIN OUTCOME MEASURES: Psychological and physical functioning at each time point was measured using the 12-item Medical Outcome Study Short-Form Health Survey (SF-12). RESULTS: Four latent classes were identified--chronic dysfunction, delayed dysfunction, recovery, and resilience. All groups had better physical health than the chronic group. Resilient and recovered individuals had greater social support and less SARS-related worry, and resilient individuals were more likely to be male. The resilient group also had greater social support than the delayed group and better physical functioning than the recovered group. CONCLUSION: This study demonstrated that longitudinal outcome trajectories following a major health-threat event in an Asian sample bear close resemblance to prototypical trajectories observed in trauma studies using Western samples. Unique predictors of the trajectories included factors observed in previous studies, such as social support, as well as factors of particular relevance to a major disease outbreak, such as SARS-related worry.