Genetic dissection of intraspecific variation in a male-specific sexual trait in Drosophila melanogaster.

An open question in evolutionary biology is the relationship between standing variation for a trait and the variation that leads to interspecific divergence. By identifying loci underlying phenotypic variation in intra- and interspecific crosses we can determine the extent to which polymorphism and divergence are controlled by the same genomic regions. Sexual traits provide abundant examples of morphological and behavioral diversity within and among species, and here we leverage variation in the Drosophila sex comb to address this question. The sex comb is an array of modified bristles or 'teeth' present on the male forelegs of several Drosophilid species. Males use the comb to grasp females during copulation, and ablation experiments have shown that males lacking comb teeth typically fail to mate. We measured tooth number in >700 genotypes derived from a multiparental advanced-intercross population, mapping three moderate-effect loci contributing to trait heritability. Two quantitative trait loci (QTLs) coincide with previously identified intra- and interspecific sex comb QTL, but such overlap can be explained by chance alone, in part because of the broad swathes of the genome implicated by earlier, low-resolution QTL scans. Our mapped QTL regions encompass 70-124 genes, but do not include those genes known to be involved in developmental specification of the comb. Nonetheless, we identified plausible candidates within all QTL intervals, and used RNA interference to validate effects at four loci. Notably, TweedleS expression knockdown substantially reduces tooth number. The genes we highlight are strong candidates to harbor segregating, functional variants contributing to sex comb tooth number.

Dynamics of the "popcorn" Wolbachia infection in outbred Aedes aegypti informs prospects for mosquito vector control.

Forty percent of the world's population is at risk of contracting dengue virus, which produces dengue fever with a potentially fatal hemorrhagic form. The wMelPop Wolbachia infection of Drosophila melanogaster reduces life span and interferes with viral transmission when introduced into the mosquito Aedes aegypti, the primary vector of dengue virus. Wolbachia has been proposed as an agent for preventing transmission of dengue virus. Population invasion by Wolbachia depends on levels of cytoplasmic incompatibility, fitness effects, and maternal transmission. Here we characterized these traits in an outbred genetic background of a potential target population of Ae. aegypti using two crossing schemes. Cytoplasmic incompatibility was strong in this background, and the maternal transmission rate of Wolbachia was high. The infection substantially reduced longevity of infected adult females, regardless of whether adults came from larvae cultured under high or low levels of nutrition or density. The infection reduced the viability of diapausing and nondiapausing eggs. Viability was particularly low when eggs were laid by older females and when diapausing eggs had been stored for a few weeks. The infection affected mosquito larval development time and adult body size under different larval nutrition levels and densities. The results were used to assess the potential for wMelPop-CLA to invade natural populations of Ae. aegypti and to develop recommendations for the maintenance of fitness in infected mosquitoes that need to compete against field insects.

Effect of mild hypothermia during cardiopulmonary bypass on erythrocytic hemoglobin oxygen delivery.

OBJECTIVES: To observe and define the degree of change in hemoglobin oxygen affinity induced by hypothermic extracorporeal circulation (ECC). DESIGN: A prospective, nonrandomized, observational study. SETTING: A single university medical center. PARTICIPANTS: Seventeen patients presenting for elective cardiac surgery. INTERVENTIONS: Systemic hypothermia during ECC. MEASUREMENTS AND MAIN RESULTS: During and after ECC, simultaneous arterial and mixed-venous whole-blood samples were obtained and immediately analyzed for gas tensions and hemoglobin saturation. Samples were obtained during the following times on ECC: initially after cardiopulmonary bypass onset during normothermia (37 degrees C), after cooling to 32 degrees C, and after rewarming to 37 degrees C. A fourth sample was obtained 10 to 20 minutes after discontinuation of cardiopulmonary bypass. Extracorporeal pump flow and thermodilution-determined cardiac output were also recorded for calculation of oxygen delivery and consumption. Mixed-venous results were used to calculate in vivo the blood gas tension at which hemoglobin was 50% saturated with oxygen (P50). There were no differences in P50 for the 17 patients by analysis of variance (ANOVA) for repeated measures with paired t-test with Bonferroni correction. Furthermore, no change in P50 was observed during the course of cooling and rewarming in any individual patient's samples. Oxygen delivery decreased after hypothermia and rewarming from mild hypothermia; oxygen consumption was decreased after rewarming and markedly increased after discontinuation from ECC. CONCLUSION: Mild hypothermia to 32 degrees C during ECC does not result in in vivo alterations in oxyhemoglobin dissociation and thus does not induce changes in oxygen delivery to peripheral tissues. However, oxygen usage appears to be markedly increased after cardiopulmonary bypass.

Microbiological monitoring of bone grafts: two years' experience at a tissue bank.

In the first two years of operation of a tissue bank, bone was processed on 63 occasions from 22 cadaveric donors and on 37 occasions from 185 living donors. A standardized protocol for microbiological sampling, culturing and interpretation of the results was developed. Semi-quantitative culture of washings of bone was performed on receipt by the tissue bank, and broth enrichment cultures of bone samples were performed at the end of processing, and again after irradiation. One bone donation was rejected because of heavy contamination with Klebsiella sp. on receipt, and contamination of six donations with Burkholderia cepacia was shown to have come from a water deionizer. Contamination of bone on receipt by the tissue bank decreased during the study period, probably related to increasing experience of staff harvesting bone. Microbiological surveillance of bone grafts protect recipients from infection, and is useful as a quality control of the process of bone banking.