A yeast surface display platform for plant hormone receptors: Toward directed evolution of new biosensors

Small-molecule biosensors have major applications in biotechnology and medicine but remain difficult to engineer. Plant hormone receptors represent an attractive platform for engineering such biosensors because their chemically induced dimerization architectures naturally decouple small-molecule sensing and sensor actuation. Rapid biosensor engineering will require quantitative high-throughput screening methods. Here we develop a yeast surface display (YSD) platform for the PYR1/HAB1 abscisic acid sensor of Arabidopsis thaliana. We extensively optimized PYR1 surface display, HAB1 purification, and binding reaction conditions. Our system reproduces previous results with wild-type and engineered receptors, and a mathematical analysis of the PYR1/HAB1 system allows us to infer all binding constants. Critically, we find that a previously engineered PYR1 receptor with altered ligand specificity binds HAB1 with identical affinity, suggesting that substantial reengineering of the PYR1 binding pocket does not compromise sensor actuation. This YSD platform for A. thaliana PYR1/HAB1 will facilitate future biosensor engineering efforts.     

Fox Hunting in Wild Apples: Searching for Novel Genes in Malus Sieversii

Malus sieversii is considered the progenitor of modern apple (Malus pumila) cultivars and to represent a valuable source of genetic diversity. Despite the importance of M. sieversii as a source of disease resistance, stress tolerance, and novel fruit traits, little is known about gene function and diversity in M. sieversii. Notably, a publicly annotated genome sequence for this species is not available. In the current study, the FOX (Full-length cDNA OvereXpressing) gene hunting system was used to construct a library of transgenic lines of Arabidopsis in which each transgenic line overexpresses a full-length gene obtained from a cDNA library of the PI619283 accession of M. sieversii. The cDNA library was constructed from mRNA obtained from bark tissues collected in late fall–early winter, a time at which many abiotic stress-adaptative genes are expressed. Over 4000 apple FOX Arabidopsis lines have been established from the pool of transgenic seeds and cDNA inserts corresponding to various Gene Ontology (GO) categories have been identified. A total of 160 inserts appear to be novel, with no or limited homology to M. pumila, Arabidopsis, or poplar. Over 1300 lines have also been screened for freezing resistance. The constructed library of transgenic lines provides a valuable genetic resource for exploring gene function and diversity in Malus sieversii. Notably, no such library of t-DNA lines currently exists for any Malus species.     

Cone Photoreceptor Loss in Light-Damaged Albino Rats

We investigated the etiology of decreased cone-driven vision in a light damage (LD) model of retinal degeneration. To induce slow, moderate degeneration, albino rats underwent low-intensity light exposure for 10 days. Electroretinography was utilized to assess physiologic function of the rod- and cone-driven retinal function in LD and control rats. Immunohistochemistry targeting cone arrestin allowed for quantification of cone density and for comparison of the decline in function. Photoreceptor loss was quantified by outer nuclear layer thickness decreases, as observed by optical coherence tomography and histology. The LD rats showed decreased rod- and cone-driven function with partial recovery 30 days after cessation of light exposure. In addition, LD rats showed decreased cone photoreceptor densities in the central retinal region compared to control rats. Our results demonstrate that the loss of cone-driven visual function induced by light damage is at least partially due to the death of cone photoreceptors.     

A compact photomicroreactor design for kinetic studies of gas‐liquid photocatalytic transformations

A compact photomicroreactor assembly consisting of a capillary microreactor and small‐scale light emitting diodes was developed for the study of reaction kinetics in the gas‐liquid photocatalytic oxidation of thiophenol to phenyl disulfide within Taylor flow. The importance of photons was convincingly shown by a suction phenomenon due to the fast consumption of oxygen. Mass transfer limitations were evaluated and an operational zone without mass transfer effects was chosen to study reaction kinetics. Effects of photocatalyst loading and light sources on the reaction performance were investigated. Reaction kinetic analysis was performed to obtain reaction orders with respect to both thiophenol and oxygen based on heterogeneous and homogeneous experimental results, respectively. The Hatta number further indicated elimination of mass transfer limitations. Reaction rate constants at different photocatalyst loadings and different photon flux were calculated. Furthermore, the advantages of this photomicroreactor assembly for studying gas‐liquid photocatalytic reaction kinetics were demonstrated as compared with batch reactors. © 2015 American Institute of Chemical Engineers AIChE J, 61: 2215–2227, 2015     

The utility of energy storage to improve the economics of wind–diesel power plants in Canada

Wind energy systems have been considered for Canada's remote communities in order to reduce their costs and dependence on diesel fuel to generate electricity. Given the high capital costs, low-penetration wind–diesel systems have been typically found not to be economic. High-penetration wind–diesel systems have the benefit of increased economies of scale, and displacing significant amounts of diesel fuel, but have the disadvantage of not being able to capture all of the electricity that is generated when the wind turbines operate at rated capacity.Two representative models of typical remote Canadian communities were created using HOMER, an NREL micro-power simulator to model how a generic energy storage system could help improve the economics of a high-penetration wind–diesel system. Key variables that affect the optimum system are average annual wind speed, cost of diesel fuel, installed cost of storage and a storage systems overall efficiency. At an avoided cost of diesel fuel of 0.30 $Cdn/kWh and current installed costs, wind generators are suitable in remote Canadian communities only when an average annual wind speed of at least 6.0 m/s is present. Wind energy storage systems become viable to consider when average annual wind speeds approach 7.0 m/s, if the installed cost of the storage system is less than 1000 $Cdn/kW and it is capable of achieving at least a 75% overall energy conversion efficiency. In such cases, energy storage system can enable an additional 50% of electricity from wind turbines to be delivered.     

Catalytic olefin polymerisation at short times: Studies using specially adapted reactors

Despite the fact that the very early stages (several tens of seconds) of catalysed olefin polymerisation processes appear negligibly short with respect to the residence time of most industrial reactors, they are critical in terms of catalyst activation, obtaining good particle morphology, and avoiding irreparable problems caused by particle overheating. The different types of reactors that have been used over the course of the past few years are discussed in this feature article. It is shown that despite the difficulties encountered in finding the perfect experimental tool for this purpose, different configurations of stopped flow reactors can be used successfully to explore different aspects of what happens to the catalyst (supported and molecular) during these critical moments of polymerisation. © 2012 Canadian Society for Chemical Engineering     

The effect of beaver ponds on the nonpoint source water quality of a stream in Southwestern Wyoming

Currant Creek, a second order stream in southwestern Wyoming, has three large complexes of beaver ponds midway along its 32 km length. To determine whether these ponds improve the quality of water flowing through them, during spring and summer of 1984 and 1985 water samples were taken upstream from, within, and downstream from the pond complexes. During periods of high flow (i.e. spring runoff), concentrations of suspended solids (SS), total phosphorus (TP), sodium hydroxide-extractable phosphorus (NaOH-P, an index of biologically available P) and total Kjeldahl nitrogen (TKN) were reduced in water flowing through the beaver ponds. During low flow, beaver ponds had less effect on these parameters. Concentrations of nitrate nitrogen (NO₃-N) were reduced during both high and low flows, while concentrations of ortho-phosphate (ortho-P) did not appear to be affected by beaver ponds. Ammonia nitrogen almost always was at the limit of detection. Regression of NaOH-P versus [SS plus ortho-P] suggested that the primary source of NaOH-P was SS. In general, SS explained a large portion of the variation in TP, TKN, and NaOH-P, and often ortho-P was significantly correlated to TP. The increase in the concentration of most parameters below the area with dam complexes appears to reflect input from bank and channel erosion, and export of SS, TP, TKN and NO₃-N from beaver dam complexes was calculated to be less than that from stream sections above or below the dams. Thus the location of dams should be considered before using them to try and improve water quality. The apparent importance of bank and channel erosion as the primary source of nutrients to Currant Creek contrasts with many watersheds in agricultural areas.     

Methods for determining operation life and reactivity depletion for space reactors with fast energy spectra

Space reactors with fast neutron energy spectrums are preferred for their compactness and high fission power density, but require a high fissile inventory. The operation life estimates of these reactors are important to mission planning. This paper examines a number of fuel depletion and neutronics code packages for determining the operation lives of two space reactors with hard fast neutron energy spectra. These are: the lithium-cooled, Sectored, Compact Reactor (SCoRe-S₁₁), and the submersion subcritical safe space (S^4) reactor, cooled with a He–Xe binary gas mixture (40 g/mol). This work investigated the code packages of Monteburns 2.0, MCNPX 2.6C and TRITON and validated their prediction with fuel depletion data for a PWR fuel bundle, with satisfactory results. The operation life predictions of the two space reactors using these code packages are compared with those calculated using a simplified method that couples MCNP5 to a burnup analysis model using the Simulink^® platform. This method considers only the 10 most probable low-Z and high-Z elements of the fission yield peaks plus ¹⁴⁹Sm, and neglects the depletion of fission products due to capture and radioactive decay. The simplified method requires significantly shorter running time and its predictions of the operation lives for the two space reactors are within 0.29–12.5% of those obtained using Monteburns 2.0 and MCNPX 2.6C code packages. This method, however, is not recommended for operation life predictions for space or commercial reactors with thermal neutron spectrums.     

Reactivity control options of space nuclear reactors

This paper compares two ex-core control options of the gas-cooled Submersion Subcritical Safe Space (S^4) reactor with a fast neutrons energy spectrum: (a) rotating BeO drums with 120° thin segments of enriched B₄C in the BeO radial reflector; and (b) sliding segments in the BeO radial reflector. Investigated are the effects on the beginning-of-life (BOL) excess reactivity, reactivity depletion rate and operation life, and the spatial neutron flux distributions and fission power profiles in the core. Also investigated is the effect of reducing the thickness of the enriched B₄C segments in the control drums on the BOL excess reactivity, when one or two of the 6 drums are stuck in the shutdown position. Reducing the thickness of the B₄C segments from 0.5 mm to 0.238 mm, with one drums stuck in the shutdown position, increases BOL cold and hot-clean excess reactivity from +$1.71 and +$0.47 to +$2.38 and +$0.89, respectively. These reactivity values are almost identical to those of the reactor with one of the six reflector segments stuck open in the shutdown position. Results also showed that the control options made little difference in the reactor performance. The power peaking in the reactor core with sliding reflector segments is slightly lower and the spatial power profiles are relatively flatter. The operation life of the reactor with a sliding reflector segments control, when operating at a nominal thermal power of 471 kW, is only 22 full power days longer than with rotating drums control.