Housekeeping in Tephritid insects: the best gene choice for expression analyses in the medfly and the olive fly.

Real-time quantitative-PCR has been a priceless tool for gene expression analyses. The reaction, however, needs proper normalization with the use of housekeeping genes (HKGs), whose expression remains stable throughout the experimental conditions. Often, the combination of several genes is required for accurate normalization. Most importantly, there are no universal HKGs which can be used since their expression varies among different organisms, tissues or experimental conditions. In the present study, nine common HKGs (RPL19, tbp, ubx, GAPDH, α-TUB, ß-TUB, 14-3-3zeta, RPE and actin3) are evaluated in thirteen different body parts, developmental stages and reproductive and olfactory tissues of two insects of agricultural importance, the medfly and the olive fly. Three software programs based on different algorithms were used (geNorm, NormFinder and BestKeeper) and gave different ranking of HKG stabilities. This confirms once again that the stability of common HKGs should not be taken for granted and demonstrates the caution that is needed in the choice of the appropriate HKGs. Finally, by estimating the average of a standard score of the stability values resulted by the three programs we were able to provide a useful consensus key for the choice of the best HKG combination in various tissues of the two insects.

A structured autopsy-based audit of 370 firearm fatalities: Contribution to inform policy decisions and the probability of the injured arriving alive at a hospital and receiving definitive care.

The objectives of this autopsy-based audit of firearm-related fatalities were to acquire data to inform policy decisions and to assess the probability of the injured arriving alive at a hospital and receiving definitive care. EVALUATED VARIABLES: Demographics; co-morbidities; location and intention of the injury; toxicology; types of firearms; Abbreviated Injury Scale; Injury Severity Score (ISS); transfer means and time; and location of death. RESULTS: Of a total of 370 fatalities, 85.7% were male. The median age was 38 (9-95) years. Suicides (47%) and assaults (45.1%) were the most common underlying intentions. The most seriously injured regions were the head (44.5%), thorax (25.7%), abdomen (10.7%), and spine (5.7%). Of the 370 total subjects, 4.9% had an ISS<16 and 59.5% had an ISS≤74; both groups were classified as potentially preventable deaths. The majority (84%) died at the scene, and only 9.8% left the emergency department alive for further treatment. Multivariate analyses documented that postmortem ISS is an independent factor that predicts the probability of the injured reaching a hospital alive and receiving definitive care. Individuals injured in greater Athens and those most seriously injured in the face, abdomen or spine had significantly greater chances of reaching a hospital alive and receiving definitive care, whereas those injured by a shotgun and the positive toxicology group were significantly less likely to. In conclusion, this study provides data to inform policy decisions, calls for a surveillance network and establishes a baseline for estimating the probability regarding the location of firearm-related deaths.