Rapid selective sweep of pre-existing polymorphisms and slow fixation of new mutations in experimental evolution of Desulfovibrio vulgaris

To investigate the genetic basis of microbial evolutionary adaptation to salt (NaCl) stress, populations of Desulfovibrio vulgaris Hildenborough (DvH), a sulfate-reducing bacterium important for the biogeochemical cycling of sulfur, carbon and nitrogen, and potentially the bioremediation of toxic heavy metals and radionuclides, were propagated under salt stress or non-stress conditions for 1200 generations. Whole-genome sequencing revealed 11 mutations in salt stress-evolved clone ES9-11 and 14 mutations in non-stress-evolved clone EC3-10. Whole-population sequencing data suggested the rapid selective sweep of the pre-existing polymorphisms under salt stress within the first 100 generations and the slow fixation of new mutations. Population genotyping data demonstrated that the rapid selective sweep of pre-existing polymorphisms was common in salt stress-evolved populations. In contrast, the selection of pre-existing polymorphisms was largely random in EC populations. Consistently, at 100 generations, stress-evolved population ES9 showed improved salt tolerance, namely increased growth rate (2.0-fold), higher biomass yield (1.8-fold) and shorter lag phase (0.7-fold) under higher salinity conditions. The beneficial nature of several mutations was confirmed by site-directed mutagenesis. All four tested mutations contributed to the shortened lag phases under higher salinity condition. In particular, compared with the salt tolerance improvement in ES9-11, a mutation in a histidine kinase protein gene lytS contributed 27% of the growth rate increase and 23% of the biomass yield increase while a mutation in hypothetical gene DVU2472 contributed 24% of the biomass yield increase. Our results suggested that a few beneficial mutations could lead to dramatic improvements in salt tolerance.     

Discovery of Power-Law Growth in the Self-Renewal of Heterogeneous Glioma Stem Cell Populations

Background

Accumulating evidence indicates that cancer stem cells (CSCs) drive tumorigenesis. This suggests that CSCs should make ideal therapeutic targets. However, because CSC populations in tumors appear heterogeneous, it remains unclear how CSCs might be effectively targeted. To investigate the mechanisms by which CSC populations maintain heterogeneity during self-renewal, we established a glioma sphere (GS) forming model, to generate a population in which glioma stem cells (GSCs) become enriched. We hypothesized, based on the clonal evolution concept, that with each passage in culture, heterogeneous clonal sublines of GSs are generated that progressively show increased proliferative ability.

Methodology/Principal Findings

To test this hypothesis, we determined whether, with each passage, glioma neurosphere culture generated from four different glioma cell lines become progressively proliferative (i.e., enriched in large spheres). Rather than monitoring self-renewal, we measured heterogeneity based on neurosphere clone sizes (#cells/clone). Log-log plots of distributions of clone sizes yielded a good fit (r>0.90) to a straight line (log(% total clones) = k*log(#cells/clone)) indicating that the system follows a power-law (y = x^k) with a specific degree exponent (k = −1.42). Repeated passaging of the total GS population showed that the same power-law was maintained over six passages (CV = −1.01 to −1.17). Surprisingly, passage of either isolated small or large subclones generated fully heterogeneous populations that retained the original power-law-dependent heterogeneity. The anti-GSC agent Temozolomide, which is well known as a standard therapy for glioblastoma multiforme (GBM), suppressed the self-renewal of clones, but it never disrupted the power-law behavior of a GS population.

Conclusions/Significance

Although the data above did not support the stated hypothesis, they did strongly suggest a novel mechanism that underlies CSC heterogeneity. They indicate that power-law growth governs the self-renewal of heterogeneous glioma stem cell populations. That the data always fit a power-law suggests that: (i) clone sizes follow continuous, non-random, and scale-free hierarchy; (ii) precise biologic rules that reflect self-organizing emergent behaviors govern the generation of neurospheres. That the power-law behavior and the original GS heterogeneity are maintained over multiple passages indicates that these rules are invariant. These self-organizing mechanisms very likely underlie tumor heterogeneity during tumor growth. Discovery of this power-law behavior provides a mechanism that could be targeted in the development of new, more effective, anti-cancer agents.     

Patterns of cyto-nuclear linkage disequilibrium in Silene latifolia: genomic heterogeneity and temporal stability

Non-random association of alleles in the nucleus and cytoplasmic organelles, or cyto-nuclear linkage disequilibrium (LD), is both an important component of a number of evolutionary processes and a statistical indicator of others. The evolutionary significance of cyto-nuclear LD will depend on both its magnitude and how stable those associations are through time. Here, we use a longitudinal population genetic data set to explore the magnitude and temporal dynamics of cyto-nuclear disequilibria through time. We genotyped 135 and 170 individuals from 16 and 17 patches of the plant species Silene latifolia in Southwestern VA, sampled in 1993 and 2008, respectively. Individuals were genotyped at 14 highly polymorphic microsatellite markers and a single-nucleotide polymorphism (SNP) in the mitochondrial gene, atp1. Normalized LD (D′) between nuclear and cytoplasmic loci varied considerably depending on which nuclear locus was considered (ranging from 0.005–0.632). Four of the 14 cyto-nuclear associations showed a statistically significant shift over approximately seven generations. However, the overall magnitude of this disequilibrium was largely stable over time. The observed origin and stability of cyto-nuclear LD is most likely caused by the slow admixture between anciently diverged lineages within the species'' newly invaded range, and the local spatial structure and metapopulation dynamics that are known to structure genetic variation in this system.     

The guanine nucleotide exchange factor (GEF) Ect2 is an oncogene in human cancer

Ect2 is an oncogene in multiple human cancers. Ect2 is aberrantly overexpressed in multiple human tumor types, often as a result of targeted amplification of the ECT2 gene as part of the 3q26 amplicon. Ect2 is important for proliferation, migration and invasion of various types of cancer cells in vitro, and for NSCLC tumorigenicity in vivo. The role of Ect2 in cellular transformation is distinct from its physiologic role in cytokinesis, and many tumor cells appear to have evolved Ect2-independent cytokinesis mechanisms. In NSCLC cells, the ability of Ect2 to support transformation is linked to its mislocalization to the cytoplasm and activation of a Rac1-Pak-Mek1,2-Erk1,2 signaling axis that is regulated through its binding to the oncogenic PKCι/Par6α complex (Fig. 4). Therefore, Ect2 and PKCι are genetically linked due to their frequent co-amplification as part of the 3q26 amplicon, and functionally and biochemically linked through formation of an oncogenic PKCι-Par6-Ect2 complex that drives transformation. Further experiments will be required to determine if Ect2 and PKCι are similarly linked in other tumors, particularly those harboring 3q26 amplification. In addition, further work is needed to elucidate the molecular mechanisms that regulate the formation, dynamics and activity of the oncogenic PKCι-Par6α-Ect2 complex. These studies hold the promise of identifying novel therapeutic approaches to cancer treatment based on inhibiting Ect2 function in cancer cells.     

Winter Distribution,Movements, and Annual Survival of Radiomarked Vancouver Canada Geese in Southeast Alaska

ABSTRACT Management of Pacific Flyway Canada geese (Branta canadensis) requires information on winter distribution of different populations. Recoveries of tarsus bands from Vancouver Canada geese (B. canadensis fulva) marked in southeast Alaska, USA, ≥4 decades ago suggested that ≥83% of the population was non-migratory and that annual adult survival was high (Ŝ = 0.836). However, recovery distribution of tarsus bands was potentially biased due to geographic differences in harvest intensity in the Pacific Flyway. Also, winter distribution of Vancouver Canada geese could have shifted since the 1960s, as has occurred for some other populations of Canada geese. Because winter distribution and annual survival of this population had not recently been evaluated, we surgically implanted very high frequency radiotransmitters in 166 adult female Canada geese in southeast Alaska. We captured Vancouver Canada geese during molt at 2 sites where adults with goslings were present (breeding areas) and 2 sites where we observed nonbreeding birds only. During winter radiotracking flights in southeast Alaska, we detected 98% of 85 females marked at breeding areas and 83% of 70 females marked at nonbreeding sites, excluding 11 females that died prior to the onset of winter radiotracking. We detected no radiomarked females in coastal British Columbia, or western Washington and Oregon, USA. Most (70%) females moved ≤30 km between November and March. Our model-averaged estimate of annual survival (Ŝ = 0.844, SE = 0.050) was similar to the estimate of annual survival of geese marked from 1956 to 1960. Likely <2% of Vancouver Canada geese that nest in southeast Alaska migrate to winter areas in Oregon or Washington where they could intermix with Canada geese from other populations in the Pacific Flyway. Because annual survival of adult Vancouver Canada geese was high and showed evidence of long-term consistency, managers should examine how reproductive success and recruitment may affect the population.     

Inter‐regional comparison of land‐use effects on stream metabolism

1. Rates of whole‐system metabolism (production and respiration) are fundamental indicators of ecosystem structure and function. Although first‐order, proximal controls are well understood, assessments of the interactions between proximal controls and distal controls, such as land use and geographic region, are lacking. Thus, the influence of land use on stream metabolism across geographic regions is unknown. Further, there is limited understanding of how land use may alter variability in ecosystem metabolism across regions. 2. Stream metabolism was measured in nine streams in each of eight regions (n = 72) across the United States and Puerto Rico. In each region, three streams were selected from a range of three land uses: agriculturally influenced, urban‐influenced, and reference streams. Stream metabolism was estimated from diel changes in dissolved oxygen concentrations in each stream reach with correction for reaeration and groundwater input. 3. Gross primary production (GPP) was highest in regions with little riparian vegetation (sagebrush steppe in Wyoming, desert shrub in Arizona/New Mexico) and lowest in forested regions (North Carolina, Oregon). In contrast, ecosystem respiration (ER) varied both within and among regions. Reference streams had significantly lower rates of GPP than urban or agriculturally influenced streams. 4. GPP was positively correlated with photosynthetically active radiation and autotrophic biomass. Multiple regression models compared using Akaike’s information criterion (AIC) indicated GPP increased with water column ammonium and the fraction of the catchment in urban and reference land‐use categories. Multiple regression models also identified velocity, temperature, nitrate, ammonium, dissolved organic carbon, GPP, coarse benthic organic matter, fine benthic organic matter and the fraction of all land‐use categories in the catchment as regulators of ER. 5. Structural equation modelling indicated significant distal as well as proximal control pathways including a direct effect of land‐use on GPP as well as SRP, DIN, and PAR effects on GPP; GPP effects on autotrophic biomass, organic matter, and ER; and organic matter effects on ER. 6. Overall, consideration of the data separated by land‐use categories showed reduced inter‐regional variability in rates of metabolism, indicating that the influence of agricultural and urban land use can obscure regional differences in stream metabolism.     

Mapping of citrullinated fibrinogen B-cell epitopes in rheumatoid arthritis by imaging surface plasmon resonance

Introduction  

Rheumatoid arthritis (RA) frequently involves the loss of tolerance to citrullinated antigens, which may play a role in pathogenicity. Citrullinated fibrinogen is commonly found in inflamed synovial tissue and is a frequent target of autoantibodies in RA patients. To obtain insight into the B-cell response to citrullinated fibrinogen in RA, its autoepitopes were systematically mapped using a new methodology.     

MAXIMUM SWIM SPEEDS OF CAPTIVE AND FREE-RANGING DELPHINIDS: CRITICAL ANALYSIS OF EXTRAORDINARY PERFORMANCE

Research into dolphin swimming historically was guided by false assumptions pertaining to maximum speed. Accurate measurements on swimming speed and duration of effort of free-ranging dolphins are rare. To examine the variance of maximum swimming speeds, nearly 2,000 speed measurements were obtained for both captive and free-ranging dolphins, including Tursiops truncatus, Pseudorca crassidens, Delphinus capensis , and Delphinus delpbis . Measurements were made from videotapes of dolphins trained to swim fast around a large pool or jumping to a maximum height, videotapes of captured wild dolphins immediately after release, and sequential aerial photographs of a school of free-ranging dolphins startled by a passing airplane. Maximum horizontal speeds for trained animals were 8.2 m/sec for T. truncatus , 8.0 m/sec for D. delphis , and 8.0 m/sec for P. crassidens . Maximum speeds for T. truncatus swimming upwards, prior to vertical leaps ranged from 8.2 to 11.2 m/sec. Wild T. truncatus demonstrated a maximum speed of 5.7 m/sec. Maximum swimming speed of free-ranging D. capensis responding to multiple passes by a low flying airplane was 6.7 m/sec. There was no evidence that the freeranging dolphins have superior swimming capabilities to captive animals. The results of this study imply that realistic maximum swimming speeds for dolphins are lower than previous reports which were based on sparse data and imprecise measurement techniques.