Trophic niche and detection of the invasive signal crayfish (Pacifastacus leniusculus) in Scotland

Aquatic invasive species are a major threat to native freshwater biodiversity. The North American signal crayfish Pacifastacus leniusculus was introduced to Great Britain during the 1970s and is now widely distributed throughout England, Wales and Scotland. First recorded in Scotland in 1995, P. leniusculus is now established at more than twenty sites. The only other introduced crayfish species present in Scotland is the white-clawed crayfish Austropotamobius pallipes. A. pallipes is restricted to only two locations in Scotland, Loch Croispol and Whitemoss Reservoir. P. leniusculus negatively impacts macrophytes, invertebrates and fish though ecological and physical processes. Additionally, P. leniusculus has displaced A. pallipes throughout much of its native range within Great Britain due to competition and disease. Consequently, the two A. pallipes populations in Scotland have a high conservation value. This PhD study aimed to improve understanding of P. leniusculus invasion success by examining trophic dynamics and to develop methodologies that could improve the detection and control of P. leniusculus populations in Scotland. Stable isotope analysis was used to determine the diet composition, trophic position and whether an ontogenetic dietary shift occurs in the Loch Ken population of P. leniusculus. Bayesian mixing models indicated that P. leniusculus in Loch Ken do exhibit an ontogenetic dietary shift. Additionally, individuals of all sizes occupied the trophic position of a predator in Loch Ken suggesting that invertebrates and fish constitute an important component of P. leniusculus diet. Stable isotope analysis was used once again to compare the isotopic niche width and diet composition of P. leniusculus populations from Loch Ken and A. pallipes populations from Loch Croispol and Whitemoss Reservoir. At the species level, A. pallipes exhibited a larger niche width than that of P. leniusculus. At the population level, the isotopic signatures of the A. pallipes populations were considerably different from each other suggesting an overestimation of A. pallipes’ niche width at species level. Results showed no dietary overlap between species and Bayesian mixing models suggested P. leniusculus and A. pallipes were consuming different resources, indicating there would be no direct competition for food resources if they were to co-occur. A plus-maze study was used to determine if P. leniusculus exhibited a preference for one of four food attractants (Oncorhynchus mykiss, P. leniusculus, beef or vegetation), which could be used to improve trapping efficiency. In the maze system, P. leniusculus exhibited no preference for any food attractant presented. This would suggest that either the maze was not a good model or food attractants would not improve trapping efficiency of P. leniusculus. Additionally, a comparative investigation into the use of gill nets as a method to control P. leniusculus was conducted. Results showed that the net type and the presence of fish entangled in the net influenced the number of P. leniusculus caught. Finally, environmental DNA (eDNA) was used and evaluated for detection of P. leniusculus. A robust quantitative Polymerase Chain Reaction (qPCR) assay and DNA extraction protocol were developed. Using the developed qPCR assay, P. leniusculus eDNA was detected in controlled aquaria conditions but not in environmental water samples collected from the field. Furthermore, the quantities of P. leniusculus eDNA declined in aquaria conditions while individuals were still present suggesting the mechanisms for eDNA release by P. leniusculus are complex. Stable isotope analysis indicates that P. leniusculus exhibit an ontogenetic dietary shift, and in each life stage, P. leniusculus function as an omnivore but occupy the trophic position of a predator. Niche width analysis revealed that the diet of P. leniusculus was less general than that observed in A. pallipes and thus diet of P. leniusculus may not be responsible for invasive success. Food attractants will not enhance trapping efficiency but nets may present a potential new method to control P. leniusculus. Similarly, eDNA presents a promising new method for rapid detection of P. leniusculus. It will not be possible to eradicate P. leniusculus in Scotland but the findings of this PhD may help prevent establishment of new populations. These results should be incorporated into future management strategies for P. leniusculus populations in Scotland and may have broader applications in Great Britain and Europe

Dioxin-Dependent and Dioxin-Independent Gene Batteries: Comparison of Liver and Kidney in AHR-Null Mice

The aryl hydrocarbon receptor (AHR) is a widely expressed ligand-dependent transcription factor that mediates cellular responses to dioxins and other planar aromatic hydrocarbons. Ahr-null mice are refractory to the toxic effects of dioxin exposure. Although some mechanistic aspects of AHR activity are well understood, the tissue specificity of AHR effects remains unclear, both during development and following administration of exogenous ligands. To address the latter issue, we defined and compared transcriptional responses to dioxin exposure in the liver and kidney of wild-type and Ahr-null adult C57BL/6J mice treated with either 2,3,7,8-tetrachlorodibenzo-p-dioxin or corn-oil vehicle. In both tissues, essentially all effects of dioxin on hepatic mRNA levels were mediated by the AHR. Although 297 genes were altered by dioxin exposure in the liver, only 17 were changed in the kidney, including a number of well-established AHR target genes. Ahr genotype had a large effect in both tissues, profoundly remodeling both the renal and hepatic transcriptomes. Surprisingly, a large number of genes were affected by Ahr genotype in both tissues, suggesting the presence of a basal AHR gene battery. Alterations of the renal transcriptome in Ahr-null animals were associated with perturbation of specific functional pathways and enrichment of specific DNA motifs. Our results demonstrate the importance of intertissue comparisons, highlight the basal role of the AHR in liver and kidney, and support a role in development or normal physiology

Searching for a signal: Environmental DNA (eDNA) for the detection of invasive signal crayfish, Pacifastacus leniusculus (Dana, 1852)

Environmental DNA (eDNA) is a rapid, non-invasive method for species detection and distribution using DNA deposited in the environment by target organisms. eDNA has become a recognised and powerful tool for detecting invasive species in a broad range of aquatic ecosystems. We examined the use of eDNA as a tool for detecting the invasive American signal crayfish Pacifastacus leniusculus in Scotland. Species-specific TaqMan probe and primers were designed for P. leniusculus and a robust quantitative PCR (qPCR) assay and DNA extraction protocol were developed. We investigated the detection capability for P. leniusculus from water samples in a controlled laboratory experiment and determined whether crayfish density (low = 1 crayfish 5.5 L-1 or high = 3 crayfish 5.5 L-1) or length of time in tanks (samples taken at 1, 3 and 7 days) influenced DNA detectability. Additionally, the persistence of DNA was investigated after P. leniusculus removal (samples taken at 1, 3 and 7 days post removal). P. leniusculus DNA was consistently detected during the entire 7-day period and higher density tanks yielded stronger positive results with lower Ct values. After removal of P. leniusculus, there was a rapid and continuous decrease in the detectability of DNA. P. leniusculus DNA could only be detected in high density tanks by the end of the 7-day period, while DNA was no longer detectable in low density tanks after 72 hours. Preliminary field experiments sampled water from three sites in winter and five sites in summer. P. leniusculus was known to be present at two of these sites. P. leniusculus was not detected at any site in winter. However, in summer, positive signals were observed at sites with known P. leniusculus, and at sites where P. leniusculus was believed to be present anecdotally, but not confirmed. All sites where crayfish were believed to be absent were negative for eDNA. Therefore, eDNA represents a promising technique to detect and monitor invasive P. leniusculus, although the presence of detectable amounts of eDNA may be season and location dependent, even where invasive crayfish are known to be present

A HIV-1 Tat mutant protein disrupts HIV-1 Rev function by targeting the DEAD-box RNA helicase DDX1

BACKGROUND: Previously we described a transdominant negative mutant of the HIV-1 Tat protein, termed Nullbasic, that downregulated the steady state levels of unspliced and singly spliced viral mRNA, an activity caused by inhibition of HIV-1 Rev activity. Nullbasic also altered the subcellular localizations of Rev and other cellular proteins, including CRM1, B23 and C23 in a Rev-dependent manner, suggesting that Nullbasic may disrupt Rev function and trafficking by intervening with an unidentified component of the Rev nucleocytoplasmic transport complex. RESULTS: To seek a possible mechanism that could explain how Nullbasic inhibits Rev activity, we used a proteomics approach to identify host cellular proteins that interact with Nullbasic. Forty-six Nullbasic-binding proteins were identified by mass spectrometry including the DEAD-box RNA helicase, DDX1. To determine the effect of DDX1 on Nullbasic-mediated Rev activity, we performed cell-based immunoprecipitation assays, Rev reporter assays and bio-layer interferometry (BLI) assays. Interaction between DDX1 and Nullbasic was observed by co-immunoprecipitation of Nullbasic with endogenous DDX1 from cell lysates. BLI assays showed a direct interaction between Nullbasic and DDX1. Nullbasic affected DDX1 subcellular distribution in a Rev-independent manner. Interestingly overexpression of DDX1 in cells not only restored Rev-dependent mRNA export and gene expression in a Rev reporter assay but also partly reversed Nullbasic-induced Rev subcellular mislocalization. Moreover, HIV-1 wild type Tat co-immunoprecipitated with DDX1 and overexpression of Tat could rescue the unspliced viral mRNA levels inhibited by Nullbasic in HIV-1 expressing cells. CONCLUSIONS: Nullbasic was used to further define the complex mechanisms involved in the Rev-dependent nuclear export of the 9 kb and 4 kb viral RNAs. All together, these data indicate that DDX1 can be sequestered by Nullbasic leading to destabilization of the Rev nucleocytoplasmic transport complex and decreased levels of Rev-dependent viral transcripts. The outcomes support a role for DDX1 in maintenance of a Rev nuclear complex that transports viral RRE-containing mRNA to the cytoplasm. To our knowledge Nullbasic is the first anti-HIV protein that specifically targets the cellular protein DDX1 to block Rev’s activity. Furthermore, our research raises the possibility that wild type Tat may play a previously unrecognized but very important role in Rev function. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12977-014-0121-9) contains supplementary material, which is available to authorized users

Reduction in BACE1 decreases body weight, protects against diet-induced obesity and enhances insulin sensitivity in mice

Insulin resistance and impaired glucose homoeostasis are important indicators of Type 2 diabetes and are early risk factors of AD (Alzheimer's disease). An essential feature of AD pathology is the presence of BACE1 (β-site amyloid precursor protein-cleaving enzyme 1), which regulates production of toxic amyloid peptides. However, whether BACE1 also plays a role in glucose homoeostasis is presently unknown. We have used transgenic mice to analyse the effects of loss of BACE1 on body weight, and lipid and glucose homoeostasis. BACE1−/− mice are lean, with decreased adiposity, higher energy expenditure, and improved glucose disposal and peripheral insulin sensitivity than wild-type littermates. BACE1−/− mice are also protected from diet-induced obesity. BACE1-deficient skeletal muscle and liver exhibit improved insulin sensitivity. In a skeletal muscle cell line, BACE1 inhibition increased glucose uptake and enhanced insulin sensitivity. The loss of BACE1 is associated with increased levels of UCP1 (uncoupling protein 1) in BAT (brown adipose tissue) and UCP2 and UCP3 mRNA in skeletal muscle, indicative of increased uncoupled respiration and metabolic inefficiency. Thus BACE1 levels may play a critical role in glucose and lipid homoeostasis in conditions of chronic nutrient excess. Therefore strategies that ameliorate BACE1 activity may be important novel approaches for the treatment of diabetes

Host pathogen interactions in relation to management of light leaf spot disease (caused by Pyrenopeziza brassicae) on Brassica species

Light leaf spot, caused by Pyrenopeziza brassicae, is currently the most damaging disease problem in oilseed rape in the UK. According to recent survey data, the severity of epidemics has increased progressively across the UK, with current yield losses of up to £160M per annum in England and more severe epidemics in Scotland. Light leaf spot is a polycyclic disease with primary inoculum consisting of air-borne ascospores produced on diseased debris from the previous cropping season. Splash-dispersed conidia produced on diseased leaves are the main component of the secondary inoculum. P. brassicae is also able to infect and cause considerable yield losses on vegetable brassicas, especially Brussels sprouts. There may be spread of light leaf spot among different brassica species. Since they have a wide host range, Pyrenopeziza brassicae populations are likely to have considerable genetic diversity and there is evidence suggesting population variations between different regions, which need further study. Available disease-management tools are not sufficient to provide adequate control of the disease. There is a need to identify new sources of resistance, which can be integrated with fungicide applications to achieve sustainable management of light leaf spot. Several major resistance genes and quantitative trait loci have been identified in previous studies, but rapid improvements in the understanding of molecular mechanisms underpinning B. napus – P. brassicae interactions can be expected through exploitation of novel genetic and genomic information for brassicas and extracellular fungal pathogens.Peer reviewe

Understanding the uncertainty in global forest carbon turnover

The length of time that carbon remains in forest biomass is one of the largest uncertainties in the global carbon cycle, with both recent historical baselines and future responses to environmental change poorly constrained by available observations. In the absence of large-scale observations, models used for global assessments tend to fall back on simplified assumptions of the turnover rates of biomass and soil carbon pools. In this study, the biomass carbon turnover times calculated by an ensemble of contemporary terrestrial biosphere models (TBMs) are analysed to assess their current capability to accurately estimate biomass carbon turnover times in forests and how these times are anticipated to change in the future. Modelled baseline 1985-2014 global average forest biomass turnover times vary from 12.2 to 23.5 years between TBMs. TBM differences in phenological processes, which control allocation to, and turnover rate of, leaves and fine roots, are as important as tree mortality with regard to explaining the variation in total turnover among TBMs. The different governing mechanisms exhibited by each TBM result in a wide range of plausible turnover time projections for the end of the century. Based on these simulations, it is not possible to draw robust conclusions regarding likely future changes in turnover time, and thus biomass change, for different regions. Both spatial and temporal uncertainty in turnover time are strongly linked to model assumptions concerning plant functional type distributions and their controls. Thirteen model-based hypotheses of controls on turnover time are identified, along with recommendations for pragmatic steps to test them using existing and novel observations. Efforts to resolve uncertainty in turnover time, and thus its impacts on the future evolution of biomass carbon stocks across the world\u27s forests, will need to address both mortality and establishment components of forest demography, as well as allocation of carbon to woody versus non-woody biomass growth

Screening for Syphilis Infection in Nonpregnant Adults and Adolescents: US Preventive Services Task Force Recommendation Statement

Clinical Review & Education US Preventive Services Task Force | RECOMMENDATION STATEMENT Screening for Syphilis Infection in Nonpregnant Adults and Adolescents US Preventive Services Task Force Recommendation Statement US Preventive Services Task Force (USPSTF) Editorial page 2281 IMPORTANCE In 2014, 19 999 cases of syphilis were reported in the United States. Left untreated, syphilis can progress to late-stage disease in about 15% of persons who are infected. Late-stage syphilis can lead to development of inflammatory lesions throughout the body, which can lead to cardiovascular or organ dysfunction. Syphilis infection also increases the risk for acquiring or transmitting HIV infection. OBJECTIVE To update the 2004 US Preventive Services Task Force (USPSTF) recommendation on screening for syphilis infection in nonpregnant adults. Screening for syphilis in pregnant women was updated in a separate recommendation statement in 2009 (A recommendation). EVIDENCE REVIEW The USPSTF reviewed the evidence on screening for syphilis infection in asymptomatic, nonpregnant adults and adolescents, including patients coinfected with other sexually transmitted infections (such as HIV). Author Audio Interview at jama.com Related article page 2328 and JAMA Patient Page page 2367 CME Quiz at jamanetworkcme.com and CME Questions page 2342 Related articles at jamadermatology.com, jamaneurology.com, jamapediatrics.com FINDINGS The USPSTF found convincing evidence that screening for syphilis infection in asymptomatic, nonpregnant persons at increased risk for infection provides substantial benefit. Accurate screening tests are available to identify syphilis infection in populations at increased risk. Effective treatment with antibiotics can prevent progression to late-stage disease, with small associated harms, providing an overall substantial health benefit. CONCLUSIONS AND RECOMMENDATION The USPSTF recommends screening for syphilis infection in persons who are at increased risk for infection. (A recommendation) Authors/Group Information: The USPSTF members are listed at the end of the article. JAMA. 2016;315(21):2321-2327. doi:10.1001/jama.2016.5824 Corresponding Author: Kirsten Bibbins-Domingo, PhD, MD, MAS ([email protected]). T he US Preventive Services Task Force (USPSTF) makes recommendations about the effectiveness of specific preventive care services for patients without obvious related signs or symptoms. It bases its recommendations on the evidence of both the benefits and harms of the service and an assessment of the bal- ance. The USPSTF does not consider the costs of providing a ser- vice in this assessment. The USPSTF recognizes that clinical decisions involve more con- siderations than evidence alone. Clinicians should understand the evidence but individualize decision making to the specific patient or situation. Similarly, the USPSTF notes that policy and coverage decisions involve considerations in addition to the evidence of clini- cal benefits and harms. Summary of Recommendation and Evidence The USPSTF recommends screening for syphilis infection in per- sons who are at increased risk for infection. (A recommendation) (Figure 1) jama.com See the Clinical Considerations section later in this article for in- formation on risk factors for infection. Rationale Importance The number of cases of primary and secondary syphilis have been in- creasing since 2000. In 2014, 19 999 cases (6.3 cases per 100 000 persons)ofprimaryandsecondarysyphiliswerereportedintheUnited States. 1 Left untreated, syphilis can progress to late-stage disease in approximately 15% of persons who are infected. 2 Consequences of late-stage syphilis include development of inflammatory lesions throughout the body (eg, aortitis, gummatous lesions, and osteitis), which can lead to cardiovascular or organ dysfunction. Syphilis in- fection of the central nervous system (neurosyphilis) can occur at any stage of disease and can result in blindness, paresis, tabes dor- salis, and dementia. Syphilis infection also increases the risk for ac- quiring or transmitting HIV infection. The USPSTF addresses screening for syphilis in pregnant women in a separate recommendation statement. 3 (Reprinted) JAMA June 7, 2016 Volume 315, Number 21 Copyright 2016 American Medical Association. All rights reserved. Downloaded From: http://jamanetwork.com/ by a University of California - Los Angeles User on 09/21/201