The white gene and locomotor recovery from anoxia in Drosophila melanogaster

Locomotor recovery from anoxia is complicated and little is known about the molecular and cellular mechanisms regulating anoxic recovery in Drosophila. For this thesis I established a protocol for large-scale analysis of locomotor activity in adult flies with exposure to a transient anoxia. Using this protocol I observed that wild-type Canton-S flies recovered faster and more consistently from anoxia than the white-eyed mutant w1118, which carries a null allele of w1118 in an isogenic genetic background. Both Canton-S and w1118 are commonly used controls in the Drosophila community. Genetic analysis including serial backcrossing, RNAi knockdown, w+ duplication to Y chromosome as well as gene mutation revealed a strong association between the white gene and the timing of locomotor recovery. I also found that the locomotor recovery phenotype is independent of white-associated eye pigmentation, that heterozygous w+ allele was haplo-insufficient to induce fast and consistent locomotor recovery from anoxia in female flies, and that mini-white is insufficient to promote fast and consistent locomotor recovery. Moreover, locomotor recovery was delayed in flies with RNAi knockdown of white in subsets of serotonin neurons in the central nervous system. I further demonstrated that mutations of phosphodiesterase genes (PDE) displayed wild-type-like fast and consistent locomotor recovery, and that locomotor recovery was light-sensitive in the night in w1118. The delayed locomotor recovery and the light sensitivity were eliminated in PDE mutants that were dual-specific or cyclic guanosine monophosphate (cGMP)-specific. Up-regulation of cGMP using multiple approaches including PDE mutation, sildenafil feeding or specific expression of an atypical soluble guanylyl cyclase (Gyc88E) was sufficient to suppress w-RNAi induced delay of locomotor recovery. Taken together, these data strongly support the hypothesis that White transports cGMP and promotes fast and consistent locomotor recovery from anoxia.

Examination of the Effect of Time Delay on Fragmentation

The main objective of blasting is to produce optimum fragmentation for downstream processing. Fragmentation is usually considered optimum when the average fragment size is minimum and the fragmentation distribution as uniform as possible. One of the parameters affecting blasting fragmentation is believed to be time delay between holes of the same row. Although one can find a significant number of studies in the literature, which examine the relationship between time delay and fragmentation, their results have been often controversial. The purpose of this work is to increase the level of understanding of how time delay between holes of the same row affects fragmentation. Two series of experiments were conducted for this purpose. The first series involved tests on small scale grout and granite blocks to determine the moment of burden detachment. The instrumentation used for these experiments consisted mainly of strain gauges and piezoelectric sensors. Some experiments were also recorded with a high speed camera. It was concluded that the time of detachment for this specific setup is between 300 and 600 μs. The second series of experiments involved blasting of a 2 meter high granite bench and its purpose was the determination of the hole-to-hole delay that provides optimum fragmentation. The fragmentation results were assessed with image analysis software. Moreover, vibration was measured close to the blast and the experiments were recorded with high speed cameras. The results suggest that fragmentation was optimum when delays between 4 and 6 ms were used for this specific setup. Also, it was found that the moment at which gases first appear to be venting from the face was consistently around 6 ms after detonation.