Murine Model of Hindlimb Ischemia

In the United States, peripheral arterial disease (PAD) affects about 10 million individuals, and is also prevalent worldwide. Medical therapies for symptomatic relief are limited. Surgical or endovascular interventions are useful for some individuals, but long-term results are often disappointing. As a result, there is a need for developing new therapies to treat PAD. The murine hindlimb ischemia preparation is a model of PAD, and is useful for testing new therapies. When compared to other models of tissue ischemia such as coronary or cerebral artery ligation, femoral artery ligation provides for a simpler model of ischemic tissue. Other advantages of this model are the ease of access to the femoral artery and low mortality rate

Translocation as a Population Restoration Technique for Northern Bobwhites: A Review and Synthesis

Northern bobwhite (Colinus virginianus) abundance has declined precipitously for decades across much of the species range, to the point of widespread local, regional, and statewide extirpation. Because of successful translocations of other gallinaceous birds, bobwhite enthusiasts increasingly call for use of the approach. Consequently, the National Bobwhite Technical Committee (NBTC), on behalf of state agencies, requested a review and recommendation by the NBTC Science Subcommittee. Thus, our paper is co-authored by invited experts and includes reviews of peer-reviewed publications, manuscripts in these proceedings, state agency reports, experience by co-authors, and a survey of perspectives on translocations by state wildlife agency members of the NBTC. We discuss the state of science on key aspects of bobwhite conservation, offer best management practices (BMPs) for using translocation as a potential bobwhite restoration technique, and suggest ways to reduce uncertainty about implementation. We note that although conservationists operate on a relatively solid foundation of improving bobwhite abundance via increased quantity, connectivity, and quality of habitat, population restoration success to- date is relatively rare and unpredictable. Similarly, some past translocations have been unreliable with an abundance of failures and inadequate experimental designs. We conclude that because of major uncertainties regarding habitat, population phenomena (e.g., Allee effect) and restoration techniques, outcomes of translocations remain unpredictable; thus, future efforts must be a part of sound and rigorous peer-reviewed research. To improve scientific efforts, we recommend the following BMPs for future translocations: (1) target bobwhite abundance should be \u3e800 post-translocation which will likely necessitate ≥600 ha of suitable and accessible habitat while a larger (e.g., \u3e800 ha) area will be needed in areas with lower carrying capacity and when sites are highly fragmented or isolated, (2) personnel should identify and avoid stressors to bobwhites in all phases of the translocation process (i.e., capture, holding, transportation, and release), (3) source populations should be disease free and from similar environments and latitude; preferably from the nearest suitable source, (4) conspecifics should be present on recipient sites (5) birds should be released just before the breeding season (i.e., March or April), and (6) the translocation should incorporate robust short- and long-term bird (i.e., abundance and/or density) and habitat monitoring efforts (i.e., the Coordinated Implementation Program (CIP) of the National Bobwhite Conservation Initiative (NBCI)). In conclusion, we note that translocation of bobwhites is not a panacea for broad scale restoration of bobwhites; however, the technique should remain at the forefront of bobwhite science, taking into account knowledge of the species’ life history and ecology, so that a practical and reliable solution can be developed. We recognize this paper is just the beginning of vigorous debate, testing of concepts, and on-the ground implementation of successful bobwhite conservation

The complete genome sequence of the phytopathogenic fungus Sclerotinia sclerotiorum reveals insights into the genome architecture of broad host range pathogens

Sclerotinia sclerotiorum is a phytopathogenic fungus with over 400 hosts including numerous economically important cultivated species. This contrasts many economically destructive pathogens that only exhibit a single or very few hosts. Many plant pathogens exhibit a “two-speed” genome. So described because their genomes contain alternating gene rich, repeat sparse and gene poor, repeat-rich regions. In fungi, the repeat-rich regions may be subjected to a process termed repeat-induced point mutation (RIP). Both repeat activity and RIP are thought to play a significant role in evolution of secreted virulence proteins, termed effectors. We present a complete genome sequence of S. sclerotiorum generated using Single Molecule Real-Time Sequencing technology with highly accurate annotations produced using an extensive RNA sequencing data set. We identified 70 effector candidates and have highlighted their in planta expression profiles. Furthermore, we characterized the genome architecture of S. sclerotiorum in comparison to plant pathogens that exhibit “two-speed” genomes. We show that there is a significant association between positions of secreted proteins and regions with a high RIP index in S. sclerotiorum but we did not detect a correlation between secreted protein proportion and GC content. Neither did we detect a negative correlation between CDS content and secreted protein proportion across the S. sclerotiorum genome. We conclude that S. sclerotiorum exhibits subtle signatures of enhanced mutation of secreted proteins in specific genomic compartments as a result of transposition and RIP activity. However, these signatures are not observable at the whole-genome scale

Awake brain tumor resection during pregnancy: Decision making and technical nuances

The co-occurrence of primary brain tumor and pregnancy poses unique challenges to the treating physician. If a rapidly growing lesion causes life-threatening mass effect, craniotomy for tumor debulking becomes urgent. The choice between awake craniotomy versus general anesthesia becomes complicated if the tumor is encroaching on eloquent brain because considerations pertinent to both patient safety and oncological outcome, in addition to fetal wellbeing, are involved. A 31-year-old female at 30 weeks gestation with twins presented to our hospital seeking awake craniotomy to resect a 7 × 6 × 5 cm left frontoparietal brain tumor with 7 mm left-to-right subfalcine herniation on imaging that led to word finding difficulty, dysfluency, right upper extremity paralysis, and right lower extremity weakness. She had twice undergone tumor debulking under general anesthesia during the same pregnancy at an outside hospital at 16 weeks and 28 weeks gestation. There were considerations both for and against awake brain tumor resection over surgery under general anesthesia. The decision-making process and the technical nuances related to awake brain tumor resection in this neurologically impaired patient are discussed. Awake craniotomy benefits the patient who harbors a tumor that encroaches on the eloquent brain by allowing a greater extent of resection while preserving the language and sensorimotor function. It can be successfully done in pregnant patients who are neurologically impaired. The patient should be motivated and well informed of the details of the process. A multidisciplinary and collaborative effort is also crucial

A REGIONAL EXPERIMENT TO EVALUATE EFFECTS OF FIRE AND FIRE SURROGATE TREATMENTS IN THE SAGEBRUSH BIOME

SageSTEP is a comprehensive regional experiment that provides critical information to managers faced with a sagebrush steppe ecosystem that is increasingly at risk from wildfire, invasive plants, and climate change. The experiment provides managers with information that can be used to restore ecological communities across the 100+ million acres of the sagebrush biome. It is designed to match the temporal and spatial scales at which managers operate, is intended to reduce management risk and uncertainty of catastrophic wildfire to the greatest degree possible, and provides managers with information that allows them to better understand tradeoffs inherent in the choice of management alternatives. The project has several features that make it ideal for testing hypotheses from state‐andtransition theory, and for discovering information that can be directly applied in a management context ‐‐ it is long‐term, experimental, multisite, multivariate, and treatments are applied across condition gradients, allowing for potential identification of biotic thresholds. The project is designed to distinguish communities that have conditions that will allow them to recover on their own following fuel or restoration treatments, versus communities that have crossed biotic thresholds, and will therefore require more expensive active restoration. SageSTEP is designed as a long‐term study, such that measurements are planned for at least 10 years after treatment implementation, or through the 2018 field season. This final report therefore describes the short‐term effects of treatments, 2‐4 years after treatment implementation., or through the 2010 field season. The Joint Fire Science Program generously funded SageSTEP for its first six years, and this funding was crucial for building an infrastructure that has now set the stage for an unprecedented long‐term study that will provide badly needed information on sagebrush steppe restoration and fuel treatment effectiveness. The infrastructure we’ve built consists of the following eight features: 1. A network of 18 sites distributed across the Great Basin, Snake River Basin, and Columbia Basin, 11 sites in a replicated woodland experiment, and 7 sites in a replicated sage‐cheat experiment (Figure 1). Each site is equivalent to a statistical block consisting of an unmanipulated control, and a set of fire and fire surrogate treatments. 2. A network of weather and soil moisture stations distributed along with the sites, that provides information on inter‐annual and geographic variation in moisture and temperature, and that is being used to interpret patterns of ecological response. 3. A small by efficient staff, consisting of scientists and technicians, responsible for continued monitoring of ecological variables through time, and maintenance of the projects’ infrastructure. 4. A funding stream from several agency sources, with current resources adequate to run the project for at least three more years, and with agreements in place to fund the project through fiscal year 2015. 5. A web of partnerships among managers, scientists, students, stakeholders, and policymakers that has worked together to design the study, implement the treatments, and learn about how sagebrush steppe system respond to alternative restoration treatments. 6. A highly effective and influential outreach program, anchored by a popular website, designed to interpret and deliver scientific information collected by SageSTEP scientists, and to distribute other relevant information originating from outside the project. 7. An on‐line database, called the SageSTEP Data Store, that offers fully proofed and validated data to analysts working within SageSTEP, and which will eventually provide the same information to other interested users. 8. The Great Basin NEON Site, NSF’s atmospheric sampling station that will soon be built at the SageSTEP Onaqui site. This link with NSF provides SageSTEP with leverage for established additional vegetation and soil monitoring facilities at Onaqui. Over the past three years, since post‐treatment data collection commenced, SageSTEP has produced a considerable amount of information, most of it now published in a total of 32 scientific papers. Key outreach products include: ● Active web site (sagestep.org), anchoring a comprehensive outreach program ● User\u27s Guides for Western Juniper & Pinyon‐Juniper woodlands ● Two Fuel Guides, one each for pre‐treatment and post‐treatment conditions ● 15 quarterly newsletters ● Six manager workshops ● 11 tours or field trips ● Three national conference symposia, consisting of 24 papers (2 symposia planned) ● 57 contributed papers at conferences ● Seven Master’s Theses and two Ph.D. Dissertations ● 15 papers published in proceedings or reports ● Ten papers published in peer‐reviewed journals (17 papers currently in review

Valuing breastfeeding: a qualitative study of women's experiences of a financial incentive scheme for breastfeeding

BACKGROUND: A cluster randomised controlled trial of a financial incentive for breastfeeding conducted in areas with low breastfeeding rates in the UK reported a statistically significant increase in breastfeeding at 6-8 weeks. In this paper we report an analysis of interviews with women eligible for the scheme, exploring their experiences and perceptions of the scheme and its impact on breastfeeding to support the interpretation of the results of the trial. METHODS: Semi-structured interviews were carried out with 35 women eligible for the scheme during the feasibility and trial stages. All interviews were recorded and verbatim transcripts analysed using a Framework Analysis approach. RESULTS: Women reported that their decisions about infant feeding were influenced by the behaviours and beliefs of their family and friends, socio-cultural norms and by health and practical considerations. They were generally positive about the scheme, and felt valued for the effort involved in breastfeeding. The vouchers were frequently described as a reward, a bonus and something to look forward to, and helping women keep going with their breastfeeding. They were often perceived as compensation for the difficulties women encountered during breastfeeding. The scheme was not thought to make a difference to mothers who were strongly against breastfeeding. However, women did believe the scheme would help normalise breastfeeding, influence those who were undecided and help women to keep going with breastfeeding and reach key milestones e.g. 6 weeks or 3 months. CONCLUSIONS: The scheme was acceptable to women, who perceived it as rewarding and valuing them for breastfeeding. Women reported that the scheme could raise awareness of breastfeeding and encourage its normalisation. This provides a possible mechanism of action to explain the results of the trial. TRIAL REGISTRATION: The trial is registered with the ISRCTN registry, number 44898617 , https://www.isrctn.com

A population of gamma-ray emitting globular clusters seen with the Fermi Large Area Telescope

Globular clusters with their large populations of millisecond pulsars (MSPs) are believed to be potential emitters of high-energy gamma-ray emission. Our goal is to constrain the millisecond pulsar populations in globular clusters from analysis of gamma-ray observations. We use 546 days of continuous sky-survey observations obtained with the Large Area Telescope aboard the Fermi Gamma-ray Space Telescope to study the gamma-ray emission towards 13 globular clusters. Steady point-like high-energy gamma-ray emission has been significantly detected towards 8 globular clusters. Five of them (47 Tucanae, Omega Cen, NGC 6388, Terzan 5, and M 28) show hard spectral power indices $(0.7 < \Gamma <1.4)$ and clear evidence for an exponential cut-off in the range 1.0-2.6 GeV, which is the characteristic signature of magnetospheric emission from MSPs. Three of them (M 62, NGC 6440 and NGC 6652) also show hard spectral indices $(1.0 < \Gamma < 1.7)$, however the presence of an exponential cut-off can not be unambiguously established. Three of them (Omega Cen, NGC 6388, NGC 6652) have no known radio or X-ray MSPs yet still exhibit MSP spectral properties. From the observed gamma-ray luminosities, we estimate the total number of MSPs that is expected to be present in these globular clusters. We show that our estimates of the MSP population correlate with the stellar encounter rate and we estimate 2600-4700 MSPs in Galactic globular clusters, commensurate with previous estimates. The observation of high-energy gamma-ray emission from a globular cluster thus provides a reliable independent method to assess their millisecond pulsar populations that can be used to make constraints on the original neutron star X-ray binary population, essential for understanding the importance of binary systems in slowing the inevitable core collapse of globular clusters.Comment: Accepted for publication in A&A. Corresponding authors: J. Kn\"odlseder, N. Webb, B. Pancraz

Fermi Large Area Telescope Constraints on the Gamma-ray Opacity of the Universe

The Extragalactic Background Light (EBL) includes photons with wavelengths from ultraviolet to infrared, which are effective at attenuating gamma rays with energy above ~10 GeV during propagation from sources at cosmological distances. This results in a redshift- and energy-dependent attenuation of the gamma-ray flux of extragalactic sources such as blazars and Gamma-Ray Bursts (GRBs). The Large Area Telescope onboard Fermi detects a sample of gamma-ray blazars with redshift up to z~3, and GRBs with redshift up to z~4.3. Using photons above 10 GeV collected by Fermi over more than one year of observations for these sources, we investigate the effect of gamma-ray flux attenuation by the EBL. We place upper limits on the gamma-ray opacity of the Universe at various energies and redshifts, and compare this with predictions from well-known EBL models. We find that an EBL intensity in the optical-ultraviolet wavelengths as great as predicted by the "baseline" model of Stecker et al. (2006) can be ruled out with high confidence.Comment: 42 pages, 12 figures, accepted version (24 Aug.2010) for publication in ApJ; Contact authors: A. Bouvier, A. Chen, S. Raino, S. Razzaque, A. Reimer, L.C. Reye

Murine and Bovine γδ T Cells Enhance Innate Immunity against Brucella abortus Infections

γδ T cells have been postulated to act as a first line of defense against infectious agents, particularly intracellular pathogens, representing an important link between the innate and adaptive immune responses. Human γδ T cells expand in the blood of brucellosis patients and are active against Brucella in vitro. However, the role of γδ T cells in vivo during experimental brucellosis has not been studied. Here we report TCRδ−/− mice are more susceptible to B. abortus infection than C57BL/6 mice at one week post-infection as measured by splenic colonization and splenomegaly. An increase in TCRγδ cells was observed in the spleens of B. abortus-infected C57BL/6 mice, which peaked at two weeks post-infection and occurred concomitantly with diminished brucellae. γδ T cells were the major source of IL-17 following infection and also produced IFN-γ. Depletion of γδ T cells from C57BL/6, IL-17Rα−/−, and GMCSF−/− mice enhanced susceptibility to B. abortus infection although this susceptibility was unaltered in the mutant mice; however, when γδ T cells were depleted from IFN-γ−/− mice, enhanced susceptibility was observed. Neutralization of γδ T cells in the absence of TNF-α did not further impair immunity. In the absence of TNF-α or γδ T cells, B. abortus-infected mice showed enhanced IFN-γ, suggesting that they augmented production to compensate for the loss of γδ T cells and/or TNF-α. While the protective role of γδ T cells was TNF-α-dependent, γδ T cells were not the major source of TNF-α and activation of γδ T cells following B. abortus infection was TNF-α-independent. Additionally, bovine TCRγδ cells were found to respond rapidly to B. abortus infection upon co-culture with autologous macrophages and could impair the intramacrophage replication of B. abortus via IFN-γ. Collectively, these results demonstrate γδ T cells are important for early protection to B. abortus infections

Evaluation of Microorganisms Cultured from Injured and Repressed Tissue Regeneration Sites in Endangered Giant Aquatic Ozark Hellbender Salamanders

Investigation into the causes underlying the rapid, global amphibian decline provides critical insight into the effects of changing ecosystems. Hypothesized and confirmed links between amphibian declines, disease, and environmental changes are increasingly represented in published literature. However, there are few long-term amphibian studies that include data on population size, abnormality/injury rates, disease, and habitat variables to adequately assess changes through time. We cultured and identified microorganisms isolated from abnormal/injured and repressed tissue regeneration sites of the endangered Ozark Hellbender, Cryptobranchus alleganiensis bishopi, to discover potential causative agents responsible for their significant decline in health and population. This organism and our study site were chosen because the population and habitat of C. a. bishopi have been intensively studied from 1969–2009, and the abnormality/injury rate and apparent lack of regeneration were established. Although many bacterial and fungal isolates recovered were common environmental organisms, several opportunistic pathogens were identified in association with only the injured tissues of C.a. bishopi. Bacterial isolates included Aeromonas hydrophila, a known amphibian pathogen, Granulicetella adiacens, Gordonai terrae, Stenotrophomonas maltophilia, Aerococcus viridans, Streptococcus pneumoniae and a variety of Pseudomonads, including Pseudomonas aeruginosa, P. stutzeri, and P. alcaligenes. Fungal isolates included species in the genera Penicillium, Acremonium, Cladosporium, Curvularia, Fusarium, Streptomycetes, and the Class Hyphomycetes. Many of the opportunistic pathogens identified are known to form biofilms. Lack of isolation of the same organism from all wounds suggests that the etiological agent responsible for the damage to C. a. bishopi may not be a single organism. To our knowledge, this is the first study to profile the external microbial consortia cultured from a Cryptobranchid salamander. The incidence of abnormalities/injury and retarded regeneration in C. a. bishopi may have many contributing factors including disease and habitat degradation. Results from this study may provide insight into other amphibian population declines