Enabling chiral separations in discovery chemistry with open‐access chiral supercritical fluid chromatography

Work from this paper details a novel walk‐up open‐access (OA) approach to enable chiral analytical method development and preparative separation of enantiomers in early discovery chemistry using supercritical fluid chromatography (SFC). We have demonstrated the success of this OA approach using immobilized chiral stationary phases (CSPs). After screening a diverse set of racemic drug candidates, we have concluded that a simplified OA chiral SFC platform can successfully purify approximately 60% of the analysed racemates. This streamlined OA workflow enables medicinal chemists with limited expertise in chiral method development to successfully and rapidly purify enantiomers for their projects using Waters UPC² and Prep100‐SFC instrumentation.     

Characteristics of hollows and hollow‐bearing trees in semi‐arid river red gum woodland and potential limitations for hollow‐dependent wildlife

Up to 37 species of the birds and microbats inhabiting inland Australia are dependent on tree cavities for breeding or roosting. The river red gum (Eucalyptus camaldulensis), a well‐known hollow‐bearing tree species, occurs in linear semi‐arid woodland along thousands of kilometres of ephemeral river channels and is the only tree species that provides widespread, aggregated hollow resources across a landscape otherwise dominated by shrublands. Here we assess the type and quantity of hollows available along ephemeral rivers of the MacDonnell Ranges bioregion in central Australia and determine which characteristics of river red gums best predict the abundance and characteristics of different tree hollows, as first steps towards assessing the current availability of hollows in the region. Approximately a third of all river red gums sampled were hollow‐bearing, but individual trees with abundant hollows were rare. Further, 36% of hollows had an entrance ≤ 5 cm, and 37% had entrances which were 6–10 cm in diameter, whereas only 13% of hollows had an entrance diameter > 20 cm suitable for larger hollow‐using species. Large and high hollows only occurred on trees that did not display post‐disturbance resprouting. Trees with multiple and diverse hollows were rare and tended to be in advanced stages of senescence and had larger stems (82.3 ± 3.33 cm) and were taller (14.4 ± 0.53 m) compared to non‐hollow‐bearing trees (23.44 ± 1.68 cm, 8.0 ± 0.34 m). Further research is required to establish whether the current abundance of hollows and diversity of hollow types are limiting to cavity‐dependent wildlife, and to identify any threats to availability of hollows.     

Quantifying habitat impacts of natural gas infrastructure to facilitate biodiversity offsetting

Habitat degradation through anthropogenic development is a key driver of biodiversity loss. One way to compensate losses is “biodiversity offsetting” (wherein biodiversity impacted is “replaced” through restoration elsewhere). A challenge in implementing offsets, which has received scant attention in the literature, is the accurate determination of residual biodiversity losses. We explore this challenge for offsetting gas extraction in the Ustyurt Plateau, Uzbekistan. Our goal was to determine the landscape extent of habitat impacts, particularly how the footprint of “linear” infrastructure (i.e. roads, pipelines), often disregarded in compensation calculations, compares with “hub” infrastructure (i.e. extraction facilities). We measured vegetation cover and plant species richness using the line‐intercept method, along transects running from infrastructure/control sites outward for 500 m, accounting for wind direction to identify dust deposition impacts. Findings from 24 transects were extrapolated to the broader plateau by mapping total landscape infrastructure network using GPS data and satellite imagery. Vegetation cover and species richness were significantly lower at development sites than controls. These differences disappeared within 25 m of the edge of the area physically occupied by infrastructure. The current habitat footprint of gas infrastructure is 220 ± 19 km² across the Ustyurt (total ~ 100,000 km²), 37 ± 6% of which is linear infrastructure. Vegetation impacts diminish rapidly with increasing distance from infrastructure, and localized dust deposition does not conspicuously extend the disturbance footprint. Habitat losses from gas extraction infrastructure cover 0.2% of the study area, but this reflects directly eliminated vegetation only. Impacts upon fauna pose a more difficult determination, as these require accounting for behavioral and demographic responses to disturbance by elusive mammals, including threatened species. This study demonstrates that impacts of linear infrastructure in regions such as the Ustyurt should be accounted for not just with respect to development sites but also associated transportation and delivery routes.     

Backbone cyclic insulin

Backbone cyclic insulin was designed and prepared by reverse proteolysis in partial organic solvent of a single‐chain precursor expressed in yeast. The precursor contains two loops to bridge the two chains of native insulin. The cyclisation method uses Achromobacter lyticus protease and should be generally applicable to proteins with C‐terminal lysine and proximal N‐terminal. The presence of the ring‐closing bond and the native insulin disulfide patterns were documented by LC–MS peptide maps. The cyclic insulin was shown to be inert towards degradation by CPY, but was somewhat labile towards chymotrypsin. Intravenous administration of the cyclic insulin to Wistar rats showed the compounds to be equipotent to HI despite much lower insulin receptor affinity. Copyright © 2010 European Peptide Society and John Wiley & Sons, Ltd.     

An easier method to identify the individual genomic composition of allopolyploid complexes

A new method for the fast identification of the genomic composition of the cyprinid Squalius alburnoides is presented. The method is based on a length polymorphism detected in the β‐actin gene, which serves as the basis for the development of a semi‐quantitative PCR.     

Effect of cropping systems on cereal stemborers in the cool‐wet and semi‐arid ecozones of the Amhara region of Ethiopia

1 Field experiments were conducted on maize and sorghum at three locations in the Amhara state of Ethiopia to determine the effects of mixed cropping on stemborer infestation, borer natural enemies and grain yields. In the cool‐wet ecozone of western Amhara, sole maize was compared with maize intercropped with faba bean, mustard, potatoes and cowpea. In the semi‐arid ecozone of eastern Amhara, the trial was conducted on both maize and sorghum with the companion crops haricot bean, sesame, cowpea and sweet potatoes. 2 The results showed that the predominant borer species in western and eastern Amhara were, respectively, Busseola fusca and Chilo partellus. In Addis Zemen, western Amhara, maize intercropped with mustard and potatoes had significantly lower pest numbers and percent tunnelling than other intercrops and the maize monocrop during the vegetative stage. In eastern Amhara, the cropping system did not significantly affect pest densities but damage to stem, ear or heads tended to be greatest when cereals were intercropped with sweet potatoes. 3 Parasitism of C. partellus by the braconid Cotesia flavipes was greater on maize than sorghum, and on maize it was greater with sweet potatoes than in other intercrops or sole maize. Cocoon mass number per plant did not vary significantly between treatments. 4 There were significant differences between treatments in yields of both sorghum and maize (per plant and per unit area) with the lowest yields observed when they were intercropped with a tuber crop. 5 The results suggest that simultaneous planting of the crop species selected has little advantage over monocropped maize.     

Semi‐continuous in situ magnetic separation for enhanced extracellular protease production—modeling and experimental validation

In modern biotechnology proteases play a major role as detergent ingredients. Especially the production of extracellular protease by Bacillus species facilitates downstream processing because the protease can be directly harvested from the biosuspension. In situ magnetic separation (ISMS) constitutes an excellent adsorptive method for efficient extracellular protease removal during cultivation. In this work, the impact of semi‐continuous ISMS on the overall protease yield has been investigated. Results reveal significant removal of the protease from Bacillus licheniformis cultivations. Bacitracin‐functionalized magnetic particles were successfully applied, regenerated and reused up to 30 times. Immediate reproduction of the protease after ISMS proved the biocompatibility of this integrated approach. Six subsequent ISMS steps significantly increased the overall protease yield up to 98% because proteolytic degradation and potential inhibition of the protease in the medium could be minimized. Furthermore, integration of semi‐continuous ISMS increased the overall process efficiency due to reduction of the medium consumption. Process simulation revealed a deeper insight into protease production, and was used to optimize ISMS steps to obtain the maximum overall protease yield. Biotechnol. Bioeng. 2013; 110: 2161–2172. © 2013 Wiley Periodicals, Inc.     

Methodological bias in the seed bank flora holds significant implications for understanding seed bank community functions

• Persistent seed banks are a key plant regeneration strategy, buffering environmental variation to allow population and species persistence. Understanding seed bank functioning within herb layer dynamics is therefore important. However, rather than assessing emergence from the seed bank in herb layer gaps, most studies evaluate the seed bank functioning via a greenhouse census. We hypothesise that greenhouse data may not reflect seed bank‐driven emergence in disturbance gaps due to methodological differences. Failure in detecting (specialist) species may then introduce methodological bias into the ecological interpretation of seed bank functions using greenhouse data.
• The persistent seed bank was surveyed in 40 semi‐natural grassland plots across a fragmented landscape, quantifying seedling emergence in both the greenhouse and in disturbance gaps. Given the suspected interpretational bias, we tested whether each census uncovers similar seed bank responses to fragmentation.
• Seed bank characteristics were similar between censuses. Census type affected seed bank composition, with >25% of species retrieved better by either census type, dependent on functional traits including seed longevity, production and size. Habitat specialists emerged more in disturbance gaps than in the greenhouse, while the opposite was true for ruderal species. Both censuses uncovered fragmentation‐induced seed bank patterns.
• Low surface area sampling, larger depth of sampling and germination conditions cause underrepresentation of the habitat‐specialised part of the persistent seed bank flora during greenhouse censuses. Methodological bias introduced in the recorded seed bank data may consequently have significant implications for the ecological interpretation of seed bank community functions based on greenhouse data.