Phosphatidylserine (PS) normally localizes to the inner leaflet of cell membranes but becomes exposed in abnormal or apoptotic cells, signaling macrophages to ingest them. Along similar lines, it seemed possible that the removal of red cells from circulation because of normal aging or in hemolytic anemias might be triggered by PS exposure. To investigate the role of PS exposure in normal red cell aging, we usedN-hydroxysuccinimide-biotin to tag rabbit red cellsin vivo, then used phycoerythrin-streptavidin to label the biotinylated cells, and annexin V-fluorescein isothiocyanate (FITC) to detect the exposed PS. Flow cytometric analysis of these cells drawn at 10-day intervals up to 70 days after biotinylation indicated that older, biotinylated cells expose more PS. Furthermore, our data match a simple model of red cell senescence that assumes both an age-dependent destruction of senescent red cells preceded by several hours of PS exposure and a random destruction of red cells without PS exposure. By using this model, we demonstrated that the exposure of PS parallels the rate at which biotinylated red cells are removed from circulation. On the other hand, using an annexin V-FITC label and flow cytometry demonstrates that exposed PS does not cause the reduced red cell life span of patients with hemolytic anemia, with the possible exception of those with unstable hemoglobins or sickle cell anemia. Thus, in some cases PS exposure on the cell surface may signal the removal of red cells from circulation, but in other cases some other signal must trigger the sequestration of cells.
To evaluate two methods for reducing metal artifacts in computed tomography (CT)--the metal deletion technique (MDT) and the selective algebraic reconstruction technique (SART)--and compare these methods with filtered back projection (FBP) and linear interpolation (LI). The institutional review board approved this retrospective HIPAA-compliant study; informed patient consent was waived. Simulated projection data were calculated for a phantom that contained water, soft tissue, bone, and iron. Clinical projection data were obtained retrospectively from 11 consecutively identified CT scans with metal streak artifacts, with a total of 178 sections containing metal. Each scan was reconstructed using FBP, LI, SART, and MDT. The simulated scans were evaluated quantitatively by calculating the average error in Hounsfield units for each pixel compared with the original phantom. Two radiologists who were blinded to the reconstruction algorithms used qualitatively evaluated the clinical scans, ranking the overall severity of artifacts for each algorithm. P values for comparisons of the image quality ranks were calculated from the binomial distribution. The simulations showed that MDT reduces artifacts due to photon starvation, beam hardening, and motion and does not introduce new streaks between metal and bone. MDT had the lowest average error (76% less than FBP, 42% less than LI, 17% less than SART). Blinded comparison of the clinical scans revealed that MDT had the best image quality 100% of the time (95% confidence interval: 72%, 100%). LI had the second best image quality, and SART and FBP had the worst image quality. On images from two CT scans, as compared with images generated by the scanner, MDT revealed information of potential clinical importance. For a wide range of scans, MDT yields reduced metal streak artifacts and better-quality images than does FBP, LI, or SART. http://radiology.rsna.org/lookup/suppl/doi:10.1148/radiol.11101782/-/DC1.
Several recently introduced ‘new’ techniques in computed tomography—iterative reconstruction, gated cardiac CT, multiple-source, and dual-energy CT—actually date back to the early days of CT. We review the historic origins and evolution of these techniques, which may provide some insight into the latest innovations in commercial CT systems.
Translation of an mRNA is generally divided into three stages: initiation, elongation and termination. The relative rates of these steps determine both the number and position of ribosomes along the mRNA, but traditional velocity sedimentation assays for the translational status of mRNA determine only the number of bound ribosomes. We developed a procedure, termed Ribosome Density Mapping (RDM), that uses site-specific cleavage of polysomal mRNA followed by separation on a sucrose gradient and northern analysis, to determine the number of ribosomes associated with specified portions of a particular mRNA. This procedure allows us to test models for translation and its control, and to examine properties of individual steps of translation in vivo. We tested specific predictions from the current model for translational control of GCN4 expression in yeast and found that ribosomes were differentially associated with the uORFs elements and coding region under different growth conditions, consistent with this model. We also mapped ribosome density along the ORF of several mRNAs, to probe basic kinetic properties of translational steps in yeast. We found no detectable decline in ribosome density between the 5′ and 3′ ends of the ORFs, suggesting that the average processivity of elongation is very high. Conversely, there was no queue of ribosomes at the termination site, suggesting that termination is not very slow relative to elongation and initiation. Finally, the RDM results suggest that less frequent initiation of translation on mRNAs with longer ORFs is responsible for the inverse correlation between ORF length and ribosomal density that we observed in a global analysis of translation. These results provide new insights into eukaryotic translation in vivo.
1 January 2015 in Journal of Vascular and Interventional Radiology