The Repertoire and Dynamics of Evolutionary Adaptations to Controlled Nutrient-Limited Environments in Yeast

The experimental evolution of laboratory populations of microbes provides an opportunity to observe the evolutionary dynamics of adaptation in real time. Until very recently, however, such studies have been limited by our inability to systematically find mutations in evolved organisms. We overcome this limitation by using a variety of DNA microarray-based techniques to characterize genetic changes—including point mutations, structural changes, and insertion variation—that resulted from the experimental adaptation of 24 haploid and diploid cultures of Saccharomyces cerevisiae to growth in either glucose, sulfate, or phosphate-limited chemostats for ∼200 generations. We identified frequent genomic amplifications and rearrangements as well as novel retrotransposition events associated with adaptation. Global nucleotide variation detection in ten clonal isolates identified 32 point mutations. On the basis of mutation frequencies, we infer that these mutations and the subsequent dynamics of adaptation are determined by the batch phase of growth prior to initiation of the continuous phase in the chemostat. We relate these genotypic changes to phenotypic outcomes, namely global patterns of gene expression, and to increases in fitness by 5–50%. We found that the spectrum of available mutations in glucose- or phosphate-limited environments combined with the batch phase population dynamics early in our experiments allowed several distinct genotypic and phenotypic evolutionary pathways in response to these nutrient limitations. By contrast, sulfate-limited populations were much more constrained in both genotypic and phenotypic outcomes. Thus, the reproducibility of evolution varies with specific selective pressures, reflecting the constraints inherent in the system-level organization of metabolic processes in the cell. We were able to relate some of the observed adaptive mutations (e.g., transporter gene amplifications) to known features of the relevant metabolic pathways, but many of the mutations pointed to genes not previously associated with the relevant physiology. Thus, in addition to answering basic mechanistic questions about evolutionary mechanisms, our work suggests that experimental evolution can also shed light on the function and regulation of individual metabolic pathways

ECG-triggered non-contrast-enhanced MR angiography (TRANCE) versus digital subtraction angiography (DSA) in patients with peripheral arterial occlusive disease of the lower extremities

OBJECTIVE: To prospectively determine the diagnostic value of electrocardiography-triggered non-contrast-enhanced magnetic resonance angiography (TRANCE) of the lower extremities including the feet versus DSA. METHODS: All 43 patients with symptomatic peripheral arterial occlusive disease (PAOD) underwent TRANCE before DSA. Quality of MRA vessel depiction was rated by two independent radiologists on a 3-point scale. Arterial segments were graded for stenoses using a 4-point scale (grade 1: no stenosis; grade 2: moderate stenosis; grade 3: severe stenosis; grade 4: occlusion). Findings were compared with those of DSA. RESULTS: In the 731 vessel segments analysed, intra-arterial DSA revealed 283 stenoses: 33.6% moderate, 16.6% severe and 49.8% occlusions. TRANCE yielded a mean sensitivity, specificity, positive and negative predictive value and diagnostic accuracy to detect severe stenoses or occlusions of 95.6%, 97.4%, 87.2%, 99.2%, 97.1% for the thigh segments and 95.2%, 87.5%, 83.2%, 96.6%, 90.5% for the calf segments. Excellent overall image quality was observed for TRANCE in 91.4% versus 95.7% (DSA) for the thigh and in 60.7% versus 91.0% for the calves, while diagnostic quality of the pedal arteries was rated as insufficient. CONCLUSION: TRANCE achieves high diagnostic accuracy in the thigh and calf regions, whereas the pedal arteries showed limited quality