Transcription factor expression levels and environmental signals constrain transcription factor innovation

Evolutionary innovation of transcription factors frequently drives phenotypic diversification and adaptation to environmental change. Transcription factors can gain or lose connections to target genes, resulting in novel regulatory responses and phenotypes. However the frequency of functional adaptation varies between different regulators, even when they are closely related. To identify factors influencing propensity for innovation, we utilise a Pseudomonas fluorescens SBW25 strain rendered incapable of flagellar mediated motility in soft-agar plates via deletion of the flagellar master regulator (fleQ). This bacterium can evolve to rescue flagellar motility via gene regulatory network rewiring of an alternative transcription factor to rescue activity of FleQ. Previously, we have identified two members (out of 22) of the RpoN-dependent enhancer binding protein (RpoN-EBP) family of transcription factors (NtrC and PFLU1132) that are capable of innovating in this way. These two transcription factors rescue motility repeatably and reliably in a strict hierarchy – with NtrC the only route in a ∆fleQ background, and PFLU1132 the only route in a ∆fleQ∆ntrC background. However, why other members in the same transcription factor family have not been observed to rescue flagellar activity is unclear. Previous work shows that protein homology cannot explain this pattern within the protein family (RpoN-EBPs), and mutations in strains that rescued motility suggested high levels of transcription factor expression and activation drive innovation. We predict that mutations that increase expression of the transcription factor are vital to unlock evolutionary potential for innovation. Here, we construct titratable expression mutant lines for 11 of the RpoN-EBPs in P. fluorescens. We show that in five additional RpoN-EBPs (FleR, HbcR, GcsR, DctD, AauR and PFLU2209), high expression levels result in different mutations conferring motility rescue, suggesting alternative rewiring pathways. Our results indicate that expression levels (and not protein homology) of RpoN-EBPs are a key constraining factor in determining evolutionary potential for innovation. This suggests that transcription factors that can achieve high expression through few mutational changes, or transcription factors that are active in the selective environment, are more likely to innovate and contribute to adaptive gene regulatory network evolution

Reproductive Failure in UK Harbour Porpoises Phocoena phocoena : Legacy of Pollutant Exposure?

This research was supported by a Marie Curie International Outgoing Fellowship within the Seventh European Community Framework Programme (Project Cetacean-stressors, PIOF-GA-2010-276145 to PDJ and SM). Additional funding was provided through the Agreement on the Conservation of Small Cetaceans of the Baltic, North East Atlantic, Irish and North Seas (ASCOBANS) (Grants SSFA/2008 and SSFA / ASCOBANS / 2010 / 5 to SM). Analysis of Scottish reproductive and teeth samples was funded by the EC-funded BIOCET project (BIOaccumulation of persistent organic pollutants in small CETaceans in European waters: transport pathways and impact on reproduction, grant EVK3-2000-00027 to GJP), and Marine Scotland (GJP). Samples examined in this research were collected under the collaborative Cetacean Strandings Investigation Programme (http://ukstrandings.org/), which is funded by the Department for Environment, Food and Rural Affairs (Defra) and the UK’s Devolved Administrations in Scotland and Wales (http://sciencesearch.defra.gov.uk/Defaul​t.aspx?Menu=Menu&Module=More&Location=No​ne&Completed=0&ProjectID=15331) (grants to PDJ, RD). UK Defra also funded the chemical analysis under a service-level agreement with the Centre for Environment, Fisheries and Aquaculture Science (grants to RJL, JB). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Peer reviewedPublisher PD

Updated Distributional Records of Selected Kentucky Fishes

Distribution records for 15 fish species of conservation interest are included for Kentucky. Notropis dorsalis, previously undocumented in Kentucky, is reported for the first time (Mayfield Creek drainage). Also reported are state rediscoveries of three species thought to be extirpated from Kentucky: Ichthyomyzon gagei (Tennessee River drainage), Hemitremia flammea (Lower Cumberland and Green river drainages), and Hybopsis amnis (upper Cumberland River drainage). Unreported drainage records comprise nine species: Ichthyomyzon castaneus (Salt River drainage and Upper Green River drainage); Ichthyomyzon unicuspis (Salt River drainage and upper Big Sandy River drainage); Campostoma pullum (Tennessee River drainage); Umbra limi (Obion Creek drainage); Lepomis marginatus (Blood River system); Ammocrypta pellucida (Tygart’s Creek drainage); Nothonotus camurus (Kinniconick Creek drainage); Percina sciera (Salt River drainage); and Percina squamata (Laurel River system). Finally, we report range extensions of Notropis maculatus (Mayfield Creek drainage and Clarks River system) and Typhlichthys subterraneus (Cumberland River drainage). All but four species (I. unicuspis, C. pullum, N. camurus, and P. sciera) have a state conservation status designation because they are rare or have limited (e.g., peripheral) distributions in Kentucky

Reduced primary patency rate in diabetic patients after percutaneous intervention results from more frequent presentation with limb-threatening ischemia

ObjectiveAlthough patients with diabetes are at increased risk of amputation from peripheral vascular disease, excellent limb-salvage rates have been achieved with aggressive surgical revascularization. It is less clear whether patients with diabetes will fare as well as nondiabetics after undergoing percutaneous lower extremity revascularization, a modality which is becoming increasingly utilized for this disease process. This study aimed to assess differential outcomes in between diabetics and nondiabetics in lower extremity percutaneous interventions.MethodsWe retrospectively studied 291 patients with respect to patient variables, complications, and outcomes for percutaneous interventions performed for peripheral occlusive disease between 2002 and 2005. Tibial vessel run-off was assessed by angiography. Patency (assessed arterial duplex) was expressed by Kaplan-Meier method and log-rank analysis. Mean follow-up was 11.6 months (range 1 to 56 months).ResultsA total of 385 interventions for peripheral occlusive disease with claudication (52.2%), rest pain (16.4%), or tissue loss (31.4%) were analyzed, including 336 primary interventions and 49 reinterventions (mean patient age 73.9 years, 50.8% male). Comorbidities included diabetes mellitus (57.2%), chronic renal insufficiency (18.4%), hemodialysis (3.8%), hypertension (81.9%), hypercholesterolemia (57%), coronary artery disease (58%), tobacco use (63.2%). Diabetics were significantly more likely to be female (55.3% vs 40.8%), and suffer from CRI (23.5% vs 12.0%), a history of myocardial infarction (36.5% vs 18.0%), and <three-vessel tibial outflow (83.5% vs 71.8%), compared with nondiabetics, although all other comorbidities and lesion characteristics were equivalent between these groups. Overall primary patency (± SE) at 6, 12, and 18 months was 85 ± 2%, 63 ± 3% and 56 ± 4%, respectively. Patients with diabetes suffered reduced primary patency at 1 year compared with nondiabetics. For nondiabetics, primary patency was 88 ± 2%, 71 ± 4%, and 58 ± 4% at 6, 12, and 18 months, while for diabetics it was 82 ± 2%, 53 ± 4%, and 49 ± 4%, respectively (P = .05). Overall secondary patency at 6, 12, and 18 months was 88 ± 2%, 76 ± 3%, and 69 ± 3%, and did not vary by diabetes status. One-year limb salvage rate was 88.3% for patients with limb-threatening ischemia, which was also similar between diabetics and nondiabetics. While univariate analysis revealed that female gender, <three-vessel tibial outflow, and a history of tobacco use were all predictive of reduced primary patency (P < .05), none of these factors significantly impacted secondary patency or limb-salvage rate. Furthermore, only limb-threatening ischemia remained a significant predictor of outcome on multivariate analysis, suggesting that the poorer primary patency in diabetics is related primarily to their propensity to present with limb-threatening disease compared with nondiabetics.ConclusionPatients with diabetes demonstrate reduced primary patency rates after percutaneous treatment of lower extremity occlusive disease, most likely due to their advanced stage of disease at presentation. However, despite a higher reintervention rate, diabetics and others with risk factors predictive of reduced primary patency can attain equivalent short-term secondary patency and limb-salvage rates. Therefore, these patient characteristics should not be considered contraindications to endovascular therapy

Mental Health Symptom Severity in Cannabis-Using and Non-Using Veterans with probable PTSD

BACKGROUND: Posttraumatic Stress Disorder (PTSD) is a disabling illness suffered by many veterans returning from war. Some veterans believe that cannabis may be therapeutic for PTSD. The purpose of this study was to better understand the association between cannabis use and PTSD symptoms. METHODS: The study was a matched case-control cross-sectional evaluation of the psychiatric and sociocultural associations of cannabis use in veterans with probable PTSD. Patient self-report measures were examined comparing cannabis users (cases) to non-users (controls) who were case-matched on age and gender. RESULTS: Results indicated that there were no significant differences between cases and controls in mean PTSD Checklist-Civilian version (PCL-C) scores (59.2 and 59.1, respectively). There was also no association between PTSD scores and frequency of cannabis use. It was also observed that cases were more likely to be non-Caucasian, financially challenged, and unmarried. LIMITATIONS: The sample is a convenience sample of veterans being referred for a clinical assessment and, therefore, sampling biases may limit the generalizability of the results to other populations including veterans not seeking health care in the Veterans Affairs (VA) system. CONCLUSIONS: The results do not support the theory that cannabis use would be associated with less severe PTSD symptoms. Results do suggest important sociocultural differences in cannabis users compared to controls

Temporal and Spatial Blood Feeding Patterns of Urban Mosquitoes In the San Juan Metropolitan Area, Puerto Rico

Urban ecosystems are a patchwork of habitats that host a broad diversity of animal species. Insects comprise a large portion of urban biodiversity which includes many pest species, including those that transmit pathogens. Mosquitoes (Diptera: Culicidae) inhabit urban environments and rely on sympatric vertebrate species to complete their life cycles, and in this process transmit pathogens to animals and humans. Given that mosquitoes feed upon vertebrates, they can also act as efficient samplers that facilitate detection of vertebrate species that utilize urban ecosystems. In this study, we analyzed DNA extracted from mosquito blood meals collected temporally in multiple neighborhoods of the San Juan Metropolitan Area, Puerto Rico to evaluate the presence of vertebrate fauna. DNA was collected from 604 individual mosquitoes that represented two common urban species, Culex quinquefasciatus (n = 586) and Aedes aegypti (n = 18). Culex quinquefasciatus fed on 17 avian taxa (81.2% of blood meals), seven mammalian taxa (17.9%), and one reptilian taxon (0.85%). Domestic chickens dominated these blood meals both temporally and spatially, and no statistically significant shift from birds to mammals was detected. Aedes aegypti blood meals were from a less diverse group, with two avian taxa (11.1%) and three mammalian taxa (88.9%) identified. The blood meals we identified provided a snapshot of the vertebrate community in the San Juan Metropolitan Area and have potential implications for vector-borne pathogen transmission

Comparing the Vaginal Wall Sling with Autologous Rectus Fascia and Polypropylene Sling on Outcome and Patient Satisfaction

Since 2011 FDA safety update on transvaginal synthetic mesh for treatment of stress urinary incontinence (SUI) and pelvic organ prolapse (POP), providers are encouraged to counsel patients on risks related to synthetic mesh slings, such as dyspareunia, erosion, and extrusion. As an alternative to synthetic slings patients often choose autologous slings. We aim to compare outcome and patient satisfaction in patients, who received an autologous vaginal wall sling (VWS), rectus fascia sling (RFS), or synthetic suburethral sling (SSS) for treatment of SUI

Estimating the prevalence of problem drug use from drug-related mortality data.

BACKGROUND AND AIMS: Indirect estimation methods are required for estimating the size of populations where only a proportion of individuals are observed directly, such as problem drug users (PDUs). Capture-recapture and multiplier methods are widely used, but have been criticized as subject to bias. We propose a new approach to estimating prevalence of PDU from numbers of fatal drug-related poisonings (fDRPs) using linked databases, addressing the key limitations of simplistic 'mortality multipliers'. METHODS: Our approach requires linkage of data on a large cohort of known PDUs to mortality registers and summary information concerning additional fDRPs observed outside this cohort. We model fDRP rates among the cohort and assume that rates in unobserved PDUs are equal to rates in the cohort during periods out of treatment. Prevalence is estimated in a Bayesian statistical framework, in which we simultaneously fit regression models to fDRP rates and prevalence, allowing both to vary by demographic factors and the former also by treatment status. RESULTS: We report a case study analysis, estimating the prevalence of opioid dependence in England in 2008/09, by gender, age group and geographical region. Overall prevalence was estimated as 0.82% (95% credible interval = 0.74-0.94%) of 15-64-year-olds, which is similar to a published estimate based on capture-recapture analysis. CONCLUSIONS: Our modelling approach estimates prevalence from drug-related mortality data, while addressing the main limitations of simplistic multipliers. This offers an alternative approach for the common situation where available data sources do not meet the strong assumptions required for valid capture-recapture estimation. In a case study analysis, prevalence estimates based on our approach were surprisingly similar to existing capture-recapture estimates but, we argue, are based on a much more objective and justifiable modelling approach

Mechanized Metatheory for the Masses: The \u3cb\u3ePOPLMARK\u3c/b\u3e Challenge

How close are we to a world where every paper on programming languages is accompanied by an electronic appendix with machinechecked proofs? We propose an initial set of benchmarks for measuring progress in this area. Based on the metatheory of System F, a typed lambda-calculus with second-order polymorphism, subtyping, and records, these benchmarks embody many aspects of programming languages that are challenging to formalize: variable binding at both the term and type levels, syntactic forms with variable numbers of components (including binders), and proofs demanding complex induction principles. We hope that these benchmarks will help clarify the current state of the art, provide a basis for comparing competing technologies, and motivate further research