SUMMARYNon-migratory, stream-resident populations of threespine stickleback, Gasterosteus aculeatus, have a lower maximum oxygen consumption (M O2,max ) than ancestral migratory marine populations. Here, we examined laboratory-bred stream-resident and marine crosses from two locations (West and Bonsall Creeks) to determine which steps in the oxygen transport and utilization cascade evolved in conjunction with, and thus have the potential to contribute to, these differences in M O2,max . We found that West Creek stream-resident fish have larger muscle fibres (not measured in Bonsall fish), Bonsall Creek stream-resident fish have smaller ventricles, and both stream-resident populations have evolved smaller pectoral adductor and abductor muscles. However, many steps of the oxygen cascade did not evolve in stream-resident populations (gill surface area, hematocrit, mean cellular hemoglobin content and the activities of mitochondrial enzymes per gram ventricle and pectoral muscle), arguing against symmorphosis. We also studied F1 hybrids to determine which traits in the oxygen cascade have a genetic architecture similar to that of M O2,max . In West Creek, M O2,max , abductor and adductor size all showed dominance of marine alleles, whereas in Bonsall Creek, M O2,max and ventricle mass showed dominance of stream-resident alleles. We also found genetically based differences among marine populations in hematocrit, ventricle mass, pectoral muscle mass and pectoral muscle pyruvate kinase activity. Overall, reductions in pectoral muscle mass evolved in conjunction with reductions in M O2,max in both stream-resident populations, but the specific steps in the oxygen cascade that have a genetic basis similar to that of M O2,max , and are thus predicted to have the largest impact on M O2,max , differ among populations. Supplementary material available online at
The Salmoniform whole-genome duplication is hypothesized to have facilitated the evolution of anadromy, but little is known about the contribution of paralogs from this event to the physiological performance traits required for anadromy, such as salinity tolerance. Here, we determined when two candidate, salinity-responsive paralogs of the Na(+) , K(+) ATPase α subunit (α1a and α1b) evolved and studied their evolutionary trajectories and tissue-specific expression patterns. We found that these paralogs arose during a small-scale duplication event prior to the Salmoniform, but after the teleost, whole-genome duplication. The 'freshwater paralog' (α1a) is primarily expressed in the gills of Salmoniformes and an unduplicated freshwater sister species (Esox lucius) and experienced positive selection in the freshwater ancestor of Salmoniformes and Esociformes. Contrary to our predictions, the 'saltwater paralog' (α1b), which is more widely expressed than α1a, did not experience positive selection during the evolution of anadromy in the Coregoninae and Salmonine. To determine whether parallel mutations in Na(+) , K(+) ATPase α1 may contribute to salinity tolerance in other fishes, we studied independently evolved salinity-responsive Na(+) , K(+) ATPase α1 paralogs in Anabas testudineus and Oreochromis mossambicus. We found that a quarter of the mutations occurring between salmonid α1a and α1b in functionally important sites also evolved in parallel in at least one of these species. Together, these data argue that paralogs contributing to salinity tolerance evolved prior to the Salmoniform whole-genome duplication and that strong selection and/or functional constraints have led to parallel evolution in salinity-responsive Na(+) , K(+) ATPase α1 paralogs in fishes.
While the prevalence of microscopic hematuria (MH) in the general population remains high, the malignancy detection rate is low. Extensive workup may create unnecessary radiation and instrumentation. Multiple guidelines and nomograms have been published to help reduce excessive evaluations. This study aimed to review our institution's malignancy detection rate, modelled using the revised 2021 AUA/SUFU Microhematuria guideline and Kaiser-Permanente Hematuria Risk Index (HRI).METHODS: Retrospective review of our microscopic hematuria database of 3,147 patients to analyze malignancy detection rate, stratified for the 2021 AUA guidelines and HRI risk categories. Patient inclusion was RBC ! 3/hpf with completed fields for demographics, risk factors, and full work-up. Categorization of malignancy was further broken down into grade, stage, and type. Solid parenchymal tumors (SPT) were included if pathologically confirmed or highly suspicious on imaging. Patients who presented with MH but had a history of gross hematuria were stratified per AUA or HRI grouping. Statistical analysis was performed by area under receiver operating characteristic curves.RESULTS: 2,001 patients met inclusion criteria. Overall, 36 neoplasms were diagnosed after workup: 21 non-muscle invasive bladder cancer, 1 muscle-invasive bladder cancer, 2 upper tract urothelial cancers, 12 SPTs. AUA risk categories had a total positive detection rate of 0% (Low), 0.67% (Medium), and 2.83% (High). HRI score (0-11) showed malignancy detection rates of 0.97% (Low; 0-4), 2.65% (Moderate; 5-8), and 9.68% (High;(9)(10)(11). Looking only at urothelial malignancies (excluding SPT), the breakdown (AUA versus HRI respectively) was Low: 0 vs 0.18%, Medium/Moderate: 0.13 vs 2.28%, and High: 2.10 vs 9.68%. The HRI classified 1,139 patients as low-risk with 2 urothelial malignancies detected (0.18%), while the AUA classification only included 163 patients (0%). The HRI score stratification urothelial detection area under the curve for ROC was 0.812 compared to 0.709 (AUA).CONCLUSIONS: After stratifying our database population based on the 2021 AUA/SUFU Guideline and Kaiser-Permanente HRI, we found that both groups had linear increases in malignancy detection rates with increasing risk group. However, the HRI, which uses a more granular categorization, would have avoided unnecessary workup in a significantly higher proportion of patients. Future directions should analyze further sub-stratification to create a more specific nomogram that avoids unnecessary workup.
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