The complexity of proteomic instrumentation for LC-MS/MS introduces many possible sources of variability. Data-dependent sampling of peptides constitutes a stochastic element at the heart of discovery proteomics. Although this variation impacts the identification of peptides, proteomic identifications are far from completely random. In this study, we analyzed interlaboratory data sets from the NCI Clinical Proteomic Technology Assessment for Cancer to examine repeatability and
A major unmet need in LC-MS/MS-based proteomics analyses is a set of tools for quantitative assessment of system performance and evaluation of technical variability. Here we describe 46 system performance metrics for monitoring chromatographic performance, electrospray source stability, MS1 and MS2 signals, dynamic sampling of ions for MS/MS, and peptide identification. Applied to data sets from replicate LC-MS/MS analyses, these metrics displayed consistent, reasonable responses to controlled perturbations. The metrics typically displayed variations less than 10% and thus can reveal even subtle differences in performance of system components. Analyses of data from interlaboratory studies conducted under a common standard operating procedure identified outlier data and provided clues to specific causes. Moreover, interlaboratory variation reflected by the metrics indicates which system components vary the most between laboratories. Application of these metrics enables rational, quantitative quality assessment for proteomics and other LC-MS/MS analytical applications.
Optimal performance of LC-MS/MS platforms is critical toAccess to proteomics performance standards is essential for several reasons. First, to generate the highest quality data possible, proteomics laboratories routinely benchmark and perform quality control (QC) 1 monitoring of the performance of their instrumentation using standards. Second, appropriate standards greatly facilitate the development of improvements in technologies by providing a timeless standard with which to evaluate new protocols or instruments that claim to improve performance. For example, it is common practice for an individual laboratory considering purchase of a new instrument to require the vendor to run "demo" samples so that data from the new instrument can be compared head to head with existing instruments in the laboratory. Third, large scale proFrom the
Understanding how ecological conditions influence physiological responses is fundamental to forensic entomology. When determining the minimum postmortem interval with blow fly evidence in forensic investigations, using a reliable and accurate model of development is integral. Many published studies vary in results, source populations, and experimental designs. Accordingly, disentangling genetic causes of developmental variation from environmental causes is difficult. This study determined the minimum time of development and pupal sizes of three populations of Lucilia sericata Meigen (Diptera: Calliphoridae; from California, Michigan, and West Virginia) at two temperatures (20 degrees C and 33.5 degrees C). Development times differed significantly between strain and temperature. In addition, California pupae were the largest and fastest developing at 20 degrees C, but at 33.5 degrees C, though they still maintained their rank in size among the three populations, they were the slowest to develop. These results indicate a need to account for genetic differences in development, and genetic variation in environmental responses, when estimating a postmortem interval with entomological data.
Forensic entomologists rely on published developmental datasets to estimate the age of insects developing on human remains. Currently, these datasets only represent populations of targeted insects from specific locations. However, recent data indicate that populations can exhibit genetic variation in their development, including signatures of local adaptation demonstrated by regionally distinct plastic responses to their environments. In this study, three geographically distinct populations of the secondary screwworm, Cochliomyia macellaria Fabricius (Diptera: Calliphoridae; College Station, Longview, and San Marcos, TX, USA), a common blow fly collected from human remains in the southern USA, were reared in two distinct environments (cool 21 °C, 65 % relative humidity (RH); and warm 31 °C, 70 % RH) over 2 years (2011 and 2012) in order to determine differences in development time and mass. Significant differences in immature and pupal development time, as well as pupal mass, were shown to exist among strains derived from different populations and years. For immature development times, there was evidence of only an environmental effect on phenotype, while genotype by environment interactions was observed in pupal development times and pupal mass. College Station and San Marcos populations exhibited faster pupal development and smaller pupal sizes in the cooler environment relative to the Longview population, but showed an opposite trend in the warm environment. Rank order for College Station and Longview populations was reversed across years. Failure to take genetic variation into consideration when making such estimates can lead to unanticipated error and bias. These results indicate that genetics will have little impact on error when working with Texas genotypes of C. macellaria at ~30 °C and 70 % RH, but will have a more meaningful impact on error in postmortem interval estimates with this species in cooler, drier environments.
The assassination of President John Fitzgerald Kennedy (JFK) traumatized the nation. In this paper we show that evidence used to rule out a second assassin is fundamentally flawed. This paper discusses new compositional analyses of bullets reportedly to have been derived from the same batch as those used in the assassination. The new analyses show that the bullet fragments involved in the assassination are not nearly as rare as previously reported. In particular, the new test results are compared to key bullet composition testimony presented before the House Select Committee on Assassinations (HSCA). Matches of bullets within the same box of bullets are shown to be much more likely than indicated in the House Select Committee on Assassinations' testimony. Additionally, we show that one of the ten test bullets is considered a match to one or more assassination fragments. This finding means that the bullet fragments from the assassination that match could have come from three or more separate bullets. Finally, this paper presents a case for reanalyzing the assassination bullet fragments and conducting the necessary supporting scientific studies. These analyses will shed light on whether the five bullet fragments constitute three or more separate bullets. If the assassination fragments are derived from three or more separate bullets, then a second assassin is likely, as the additional bullet would not easily be attributable to the main suspect, Mr. Oswald, under widely accepted shooting scenarios [see Posner (1993), Case Closed, Bantam, New York].
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