Using the CRISPR/Cas9 system to understand neuropeptide biology and regulation

Funding was provided by a Wellcome Trust ISSF starting grant (105625/Z/14/Z), Medical Research Scotland (PhD-719-2013), GW Pharmaceuticals (PhD-719-2013 - S.5242.001) and the BBSRC (BB/J012343/1).Peer reviewedPublisher PD

Learning to inhabit the chair: knowledge transfer in contemporary Australian director training

How are theatre directors trained today? Although there is a body of work around what directors do, little sustained critical attention has been paid to the nature of teaching and learning within director training. Training a director has been called “teaching the unteachable” (Fliotsos, 2004) and leading practitioners assert “no amount of learning will make you a director” (Hall, 2000) — yet training courses do exist, and something, indeed many things, are taught in them. It is a kind of teaching and learning, though, which does not fit dominant models of pedagogy: “practical knowledge(s) […] have traditionally been hard to capture in meaningful ways and have, as a result, often been represented as a weaker alternative to theory” (Prior, 2012:xxiii). Using a theoretical framework drawn from the sociology of education, this thesis analyses how knowledge is transmitted and legitimated in creative arts training. This methodology seeks to understand the ‘on the ground’ realities of training, bringing into simultaneous view the official curriculum, institutional aspirations and the messy business of training. Using ethnographic fieldwork to apply this framework to the Directing program at the National Institute of Dramatic Art (NIDA) in Sydney, Australia, this thesis concentrates on the dual questions of what is taught and how it is taught. Research has been conducted from within the Institute, paying close, sustained attention to what goes on in the room during training. This work aims to contribute to debates around the accreditation of creative arts training, and the survival of conservatoire-style training. In a regulatory environment where institutions around the world now have to legitimate practical knowledges in systems designed to accredit primarily theoretical coursework, the framework proposed by this research allows trainers to account for the full spectrum of their work

Agaricomycetes of Ontario Tallgrass Prairies

This study provides the first characterization of the Agaricomycetes of Ontario tallgrass prairies, assesses the influence of various environmental factors, and compares results of aboveground mushroom surveys with belowground high-throughput DNA sequencing. Overall, the Mycenaceae, Ceratobasidiaceae and Polyporaceae were the most abundant, and the Clavariaceae, Entolomataceae and Sebacinaceae the richest in species. Position along a transect (geographic region) was the primary factor differentiating Agaricomycete composition of sites whereas tillage history and soil organic carbon content were secondary. The Hygrophoraceae and Clavariaceae were associated with pristine sites, and Minimedusa spp. associated with tillage. The belowground method captured most of the minor clades found aboveground and several more unique ones. The aboveground method retrieved 74 species and the belowground method 256 OTUs, with only eight shared between them

Actual and Reference Evaporative Losses and Surface Coefficients of a Maize Field during Nongrowing (Dormant) Periods

Effective water resources planning, allocation, management, and use in agroecosystems require accurate quantification of actual evapotranspiration (ETc) during growing and nongrowing (dormant) periods. Prediction of ETc for a variety of vegetation surfaces during the growing season has been researched extensively, but relatively little information exists on evaporative losses during nongrowing periods for different surfaces. The objectives of this research were to evaluate ETc in relation to available energy, precipitation, and grass and alfalfa-reference ET (ETo and ETr) for a maize (Zea mays L.) field and to analyze the dynamics of surface coefficients (Kc) during the nongrowing period (October 15–April 30). The evaporative losses were measured using a Bowen ratio energy balance system (BREBS) on an hourly basis and averaged over 24 h for three consecutive nongrowing periods: 2004–2005 (Season I), 2005–2006 (Season II), and 2006–2007 (Season III). BREBS-measured ETc was approximately 50% of available energy (Rn – G; Rn is net radiation and G is soil heat flux density) during normal and wet seasons (Seasons I and III) and 41% of available energy during a dry season (Season II). Cumulative ETc ranged from 133 mm in Season II to 167 mm in Season III and exceeded precipitation by 21% during the dry season. The ratio of ETc to precipitation was 0.85 in Season I, 1.21 in Season II, and 0.41 in Season III. ETc was approximately 50% of ETo and 36% of ETr in both Seasons I and III, whereas in Season II, ETc was 32% of ETo and 23% of ETr. Overall, measured ETc during the dormant season was generally most strongly correlated with radiation terms, particularly Rn, albedo, incoming shortwave radiation, and outgoing longwave radiation. Average surface coefficients over the three seasons were 0.44 and 0.33 for grass and alfalfa-reference surfaces, respectively. Using geometric mean Kc values to calculate ETc using a Kc ETref approach over the entire nongrowing season yielded adequate predictions with overall root mean square deviations of 0.64 and 0.67 mm day–1 for ETo and ETr, respectively. Estimates of ETc using a dual crop coefficient approach were good on a seasonal basis, but performed less well on a daily basis. Regression equations that were developed (accounting for serial autocorrelation in the ETc and ETref time series) yielded good estimates of ETc. Considering nongrowing period evaporative losses in water budget calculations would enable water regulatory agencies to better account for water use in hydrologic balance calculations over the entire year rather than only for the growing season and to better assess the progression and availability of water resources for the next growing season

Actual and Reference Evaporative Losses and Surface Coefficients of a Maize Field during Nongrowing (Dormant) Periods

Effective water resources planning, allocation, management, and use in agroecosystems require accurate quantification of actual evapotranspiration (ETc) during growing and nongrowing (dormant) periods. Prediction of ETc for a variety of vegetation surfaces during the growing season has been researched extensively, but relatively little information exists on evaporative losses during nongrowing periods for different surfaces. The objectives of this research were to evaluate ETc in relation to available energy, precipitation, and grass and alfalfa-reference ET (ETo and ETr) for a maize (Zea mays L.) field and to analyze the dynamics of surface coefficients (Kc) during the nongrowing period (October 15–April 30). The evaporative losses were measured using a Bowen ratio energy balance system (BREBS) on an hourly basis and averaged over 24 h for three consecutive nongrowing periods: 2004–2005 (Season I), 2005–2006 (Season II), and 2006–2007 (Season III). BREBS-measured ETc was approximately 50% of available energy (Rn – G; Rn is net radiation and G is soil heat flux density) during normal and wet seasons (Seasons I and III) and 41% of available energy during a dry season (Season II). Cumulative ETc ranged from 133 mm in Season II to 167 mm in Season III and exceeded precipitation by 21% during the dry season. The ratio of ETc to precipitation was 0.85 in Season I, 1.21 in Season II, and 0.41 in Season III. ETc was approximately 50% of ETo and 36% of ETr in both Seasons I and III, whereas in Season II, ETc was 32% of ETo and 23% of ETr. Overall, measured ETc during the dormant season was generally most strongly correlated with radiation terms, particularly Rn, albedo, incoming shortwave radiation, and outgoing longwave radiation. Average surface coefficients over the three seasons were 0.44 and 0.33 for grass and alfalfa-reference surfaces, respectively. Using geometric mean Kc values to calculate ETc using a Kc ETref approach over the entire nongrowing season yielded adequate predictions with overall root mean square deviations of 0.64 and 0.67 mm day–1 for ETo and ETr, respectively. Estimates of ETc using a dual crop coefficient approach were good on a seasonal basis, but performed less well on a daily basis. Regression equations that were developed (accounting for serial autocorrelation in the ETc and ETref time series) yielded good estimates of ETc. Considering nongrowing period evaporative losses in water budget calculations would enable water regulatory agencies to better account for water use in hydrologic balance calculations over the entire year rather than only for the growing season and to better assess the progression and availability of water resources for the next growing season

Reductions of lattice mKdV to qq-PVI\mathrm{P}_{VI}

This Letter presents a reduction of the lattice modified Korteweg-de-Vries equation that gives rise to a $q$-analogue of the sixth Painlev\'e equation. This new approach allows us to give the first ultradiscrete Lax representation of an ultradiscrete analogue of the sixth Painlev\'e equation.Comment: 4 page

Reaper is regulated by IAP-mediated ubiquitination

In most cases, apoptotic cell death culminates in the activation of the caspase family of cysteine proteases, leading to the orderly dismantling and elimination of the cell. The IAPs (inhibitors of apoptosis) comprise a family of proteins that oppose caspases and thus act to raise the apoptotic threshold. Disruption of IAP-mediated caspase inhibition has been shown to be an important activity for pro-apoptotic proteins in Drosophila (Reaper, HID, and Grim) and in mammalian cells (Smac/DIABLO and Omi/HtrA2). In addition, in the case of the fly, these proteins are able to stimulate the ubiquitination and degradation of IAPs by a mechanism involving the ubiquitin ligase activity of the IAP itself. In this report, we show that the Drosophila RHG proteins (Reaper, HID, and Grim) are themselves substrates for IAP-mediated ubiquitination. This ubiquitination of Reaper requires IAP ubiquitin-ligase activity and a stable interaction between Reaper and the IAP. Additionally, degradation of Reaper can be blocked by mutating its potential ubiquitination sites. Most importantly, we also show that regulation of Reaper by ubiquitination is a significant factor in determining its biological activity. These data demonstrate a novel function for IAPs and suggest that IAPs and Reaper-like proteins mutually control each other's abundance

Litigating BP\u27s Contribution Claims in Publicly Subsidized Courts: Should Contracting Parties Pay Their Own Way?

In this Article, we focus on an important problem involving mass-accident cases that was highlighted by the Deepwater Horizon litigation: overuse of courts to enforce contribution claims. These claims seek to shift incurred or expected liability and damages between the business and governmental entities that participated in the activity that gave rise to the mass-accident risk. Participants in such ventures generally have the option to determine by contract beforehand whether to subject themselves to contribution claims and, if so, whether such claims will be resolved by a publicly funded court or by a privately funded process, such as arbitration. Because the parties prosecuting and defending against contribution claims can consume judicial resources largely free of charge, it is likely they will choose to litigate in court to a greater extent than is socially desirable. We consider whether courts can effectively realign the parties\u27 incentives by charging them for the cost of using the judicial process. Taking account of the public good of judicial precedent- making, we advance a user-fee design that allows courts to waive the fee in whole or in part for contribution claims that present substantial questions of law. Analysis of the proposal\u27s application is extended generally to commercial contract disputes. Our central conclusion is that an appropriately designed user fee can effectively abate the problem of overuse without adversely affecting the functioning of the civil liability system