Interannual variation in precipitation and other planting conditions impacts seedling establishment in sown plant communities

Ecological restoration can reverse biodiversity loss worldwide, yet restoration goals and outcomes vary widely, which limits this potential. Divergent restoration outcomes may stem from variation in conditions at the outset of restoration, but empirical evidence is lacking and typically confounded with site differences. Additionally, precipitation is usually cited as the source of this variation, although a wide range of conditions can vary annually. We tested for effects of planting year on seedling establishment by installing identical restorations in three different years. Within those years, we manipulated rainfall with rain‐out shelters to disentangle the effects of precipitation from other annually variable conditions. Additionally, we tested whether increasing seed mix richness buffers against adverse planting conditions. For the first growing season after planting, we followed emergence and survival of sown prairie species and nonsown weed species to determine how planting year conditions influence an establishing plant community, if at all. We found that seedling establishment differed across planting years and precipitation treatments, and that varying emergence patterns by species led to differences in the composition of the first‐year community. We also found significant variation in sown species establishment across years when precipitation was held constant, illustrating the previously overlooked role of nonprecipitation drivers on planting year effects. Higher seed mix richness did not consistently improve establishment of sown species under different planting conditions. This research provides important experimental evidence for effects of interannual variation in planting conditions on first‐year establishment. Future work will examine how these initial changes affect longer‐term assembly dynamics.     

Genetics in non-genetic model systems

The past few decades have seen the field of genetic engineering evolve at a rapid pace, with neuroscientists now equipped with a wide range of tools for the manipulation of an animal's genome in order to study brain function. However, the number of species to which these technologies have been applied, namely the fruit fly, C. elegans, zebrafish and mouse, remains relatively few. This review will discuss the variety of approaches to genetic modification that have been developed in such traditional 'genetic systems', and highlight the progress that has been made to translate these technologies to alternative species such as rats, monkeys and birds, where certain neurobiological questions may be better studied.     

Quantifying the mechanical properties of human skin to optimise future microneedle device design

Microneedle devices are a promising minimally invasive means of delivering drugs/vaccines across or into the skin. However, there is currently a diversity of microneedle designs and application methods that have, primarily, been intuitively developed by the research community. To enable the rational design of optimised microneedle devices, a greater understanding of human skin biomechanics under small deformations is required. This study aims to develop a representative stratified model of human skin, informed by in vivo data. A multilayer finite element model incorporating the epidermis, dermis and hypodermis was established. This was correlated with a series of in-vivo indentation measurements, and the Ogden material coefficients were optimised using a material parameter extraction algorithm. The finite element simulation was subsequently used to model microneedle application to human skin before penetration and was validated by comparing these predictions with the in-vivo measurements. Our model has provided an excellent tool to predict micron-scale human skin deformation in vivo and is currently being used to inform optimised microneedle designs.     

Dynamics of carbon monoxide binding to cystathionine beta-synthase

Cystathionine beta-synthase (CBS) condenses homocysteine, a toxic metabolite, with serine in a pyridoxal phosphate-dependent reaction. It also contains a heme cofactor to which carbon monoxide (CO) or nitric oxide can bind, resulting in enzyme inhibition. To understand the mechanism of this regulation, we have investigated the equilibria and kinetics of CO binding to the highly active catalytic core of CBS, which is dimeric. CBS exhibits strong anticooperativity in CO binding with successive association constants of 0.24 and 0.02 microm(-1). Stopped flow measurements reveal slow CO association (0.0166 s(-1)) limited by dissociation of the endogenous ligand, Cys-52. Rebinding of CO and of Cys-52 following CO photodissociation were independently monitored via time-resolved resonance Raman spectroscopy. The Cys-52 rebinding rate, 4000 s(-1), is essentially unchanged between pH 7.6 and 10.5, indicating that the pK(a) of Cys-52 is shifted below pH 7.6. This effect is attributed to the nearby Arg-266 residue, which is proposed to form a salt bridge with the dissociated Cys-52, thereby inhibiting its protonation and slowing rebinding to the Fe. This salt bridge suggests a pathway for enzyme inactivation upon CO binding, because Arg-266 is located on a helix that connects the heme and pyridoxal phosphate cofactor domains.     

Fatty Acid- and Retinoid-binding Proteins Have Distinct Binding Pockets for the Two Types of Cargo

Parasitic nematodes cause serious diseases in humans, animals, and plants. They have limited lipid metabolism and are reliant on lipid-binding proteins to acquire these metabolites from their hosts. Several structurally novel families of lipid-binding proteins in nematodes have been described, including the fatty acid- and retinoid-binding protein family (FAR). In Caenorhabditis elegans, used as a model for studying parasitic nematodes, eight C. elegans FAR proteins have been described. The crystal structure of C. elegans FAR-7 is the first structure of a FAR protein, and it exhibits a novel fold. It differs radically from the mammalian fatty acid-binding proteins and has two ligand binding pockets joined by a surface groove. The first can accommodate the aliphatic chain of fatty acids, whereas the second can accommodate the bulkier retinoids. In addition to demonstrating lipid binding by fluorescence spectroscopy, we present evidence that retinol binding is positively regulated by casein kinase II phosphorylation at a conserved site near the bottom of the second pocket. far-7::GFP (green fluorescent protein) expression shows that it is localized in the head hypodermal syncytia and the excretory cell but that this localization changes under starvation conditions. In conclusion, our study provides the basic structural and functional information for investigation of inhibitors of lipid binding by FAR proteins.     

Developing a pre-entry weed risk assessment system for use in Japan

We evaluated the applicability of the Australian Weed Risk Assessment (AWRA) system in Japan. Native weeds (n = 117) and introduced plants (n = 142), whose weed status was classified by 20 plant experts, were assessed using a slightly modified version of the AWRA system designed to fit Japanese conditions. A receiver operating characteristic (ROC) curve for the system, when classifying two-thirds of the 259 taxa as weeds or non-weeds, was plotted and the area under the ROC curve was calculated. The area was 0.88 and significantly greater than 0.5. Thus, the validity of the system to classify plants was proven. The best cut-off level for the WRA score using Youden’s index was 10. When taxa whose AWRA scores were greater than 10 were regarded as weeds, the sensitivity and specificity were 0.88 and 0.78, respectively. These values were verified with the remaining one-third of the taxa. From these findings, the modified AWRA system was considered to be effective for use in Japan. However, further studies are required to set the best cut-off level in terms of maximising the benefits gained from using the system. A second screening test associated with the cut-off level also needs to be developed.