Production of the Carboxylate Reductase from Nocardia otitidiscaviarum in a Soluble,Active Form for in vitro Applications

Accessing aldehydes from carboxylate moieties is often a challenging task. In this regard, carboxylate reductases (CARs) are promising catalysts provided by nature that are able to accomplish this task in just one step, avoiding over-reduction to the alcohol product. However, the heterologous expression of CARs can be quite difficult due to the excessive formation of insoluble protein, thus hindering further characterization and application of the enzyme. Here, the heterologous production of the carboxylate reductase from Nocardia otitidiscaviarum (NoCAR) was optimized by a combination of i) optimized cultivation conditions, ii) post-translational modification with a phosphopantetheinyl transferase and iii) selection of an appropriate expression strain. Especially, the selection of Escherichia coli tuner cells as host had a strong effect on the final 110-fold increase in the specific activity of NoCAR. This highly active NoCAR was used to reduce sodium benzoate to benzaldehyde, and it was successfully assembled with an in vitro regeneration of ATP and NADPH, being capable of reducing about 30 mM sodium benzoate with high selectivity in only 2 h of reaction.     

Real-time organic aerosol chemical speciation in the indoor environment using extractive electrospray ionization mass spectrometry

Understanding the sources and composition of organic aerosol (OA) in indoor environments requires rapid measurements, since many emissions and processes have short timescales. However, real-time molecular-level OA measurements have not been reported indoors. Here, we present quantitative measurements, at a time resolution of five seconds, of molecular ions corresponding to diverse aerosol-phase species, by applying extractive electrospray ionization mass spectrometry (EESI-MS) to indoor air analysis for the first time, as part of the highly instrumented HOMEChem field study. We demonstrate how the complex spectra of EESI-MS are screened in order to extract chemical information and investigate the possibility of interference from gas-phase semivolatile species. During experiments that simulated the Thanksgiving US holiday meal preparation, EESI-MS quantified multiple species, including fatty acids, carbohydrates, siloxanes, and phthalates. Intercomparisons with Aerosol Mass Spectrometer (AMS) and Scanning Mobility Particle Sizer suggest that EESI-MS quantified a large fraction of OA. Comparisons with FIGAERO-CIMS shows similar signal levels and good correlation, with a range of 100 for the relative sensitivities. Comparisons with SV-TAG for phthalates and with SV-TAG and AMS for total siloxanes also show strong correlation. EESI-MS observations can be used with gas-phase measurements to identify co-emitted gas- and aerosol-phase species, and this is demonstrated using complementary gas-phase PTR-MS observations.     

Historical and contemporary movement and survival rates of walleye (Sander vitreus) in Lake Winnipeg,Canada

Understanding patterns of fish movement in large lake ecosystems is essential for determining appropriate management actions as differences in movement behaviour can influence life history traits such as growth and survival. Lake Winnipeg in Manitoba, Canada supports the 2nd largest walleye (Sander vitreus) commercial fishery in North America. We used mark-recapture models to determine movement and estimate survival of walleye between basins of Lake Winnipeg in historical and contemporary contexts, comparing a tag-recovery study completed historically during 1974–1977 with a contemporary (2017–2019) acoustic telemetry study. Mark-recapture models revealed comparably low but detectable annual transitions between basins from historical (0.3–1.2%) and contemporary datasets (7–8.5%). Historically, fish > 300 mm more frequently moved in a south to north direction. Contemporary estimates suggest similar length-based directionality in that fish > 350 mm were always more likely to move in a south-north direction. Contemporary annual survival derived from mark-recapture models ranged between 27 and 45% and 64.3% when derived from catch curve analysis, while independently derived annual historical survival estimates ranged between 50 and 69% and 45.5% from catch curve analysis. Using the contemporary dataset, we also observed seasonal variation in movement and survival between basins, with the greatest movement across the lake occurring during the fall. Our results demonstrate a persisting pattern of low but measurable movement, suggesting between basin movement is not unusual for Lake Winnipeg. Further, low walleye survival rates reported here for the two time periods studied, support recent management actions to reduce fishing pressure across the lake.     

Hierarchical Coherent Phonons in a Superatomic Semiconductor

The coupling of phonons to electrons and other phonons plays a defining role in material properties, such as charge and energy transport, light emission, and superconductivity. In atomic solids, phonons are delocalized over the 3D lattice, in contrast to molecular solids where localized vibrations dominate. Here, a hierarchical semiconductor that expands the phonon space by combining localized 0D modes with delocalized 2D and 3D modes is described. This material consists of superatomic building blocks (Re₆Se₈) covalently linked into 2D sheets that are stacked into a layered van der Waals lattice. Using transient reflectance spectroscopy, three types of coherent phonons are identified: localized 0D breathing modes of isolated superatom, 2D synchronized twisting of superatoms in layers, and 3D acoustic interlayer deformation. These phonons are coupled to the electronic degrees of freedom to varying extents. The presence of local phonon modes in an extended crystal opens the door to controlling material properties from hierarchical phonon engineering.     

Flow and spawning habitat relationships for Dolly Varden: Understanding habitat–population dynamics in the Canadian Western Arctic

Northern form Dolly Varden (Salvelinus malma malma) have been designated as a species of Special Concern in Canada due to declines in population abundance and potential threats. Concern over detrimental effects of low flows on population abundance prompted research on how variability in discharge regimes influence habitat availability. Habitat suitability indices for prespawning and spawning adult anadromous Dolly Varden from two streams were integrated into a two‐dimensional hydrodynamic habitat model to assess the effect of flow variability on usable habitat. Regional hydrographs were used to identify an ecologically relevant range of flows that provided optimal spawning habitat for these populations and examine the relationship between abundance and discharge. Adults spawned in the tail end of pools at moderate water depths and water velocities, and used pebble‐ to cobble‐sized substrate for building redds; whereas, prespawning adults occupied deeper pools with moderate velocities and used cobble for cover. Model outputs showed that spawning habitat availability was optimized at flow rates between 1.6 and 3.0 m³/s and between 1.0 and 6.0 m³/s in Fish Hole Creek (FHC) and Little Fish Creek, respectively. A positive relationship between flows during the fall spawning period and abundance of the FHC population suggests that higher flows coinciding with optimal habitat availability may have contributed to positive recruitment. To strengthen and refine this habitat–population relationship for Dolly Varden in this area requires investigation of a broader suite of variables associated with environmental regimes and physical habitat in reaches used for spawning.     

A practical assessment of risk-averse approaches in production lot-sizing problems

ABSTRACT

This paper presents an empirical assessment of four state-of-the-art risk-averse approaches to deal with the capacitated lot-sizing problem under stochastic demand. We analyse two mean-risk models based on the semideviation and on the conditional value-at-risk risk measures, and alternate first and second-order stochastic dominance approaches. The extensive computational experiments based on different instances characteristics and on a case-study suggest that CVaR exhibits a good trade-off between risk and performance, followed by the semideviation and first-order stochastic dominance approach. For all approaches, enforcing risk-aversion helps to reduce the cost-standard deviation substantially, which is usually accomplished via increasing production rates. Overall, we can say that very risk-averse decision-makers would be willing to pay an increased price to have a much less risky solution given by CVaR. In less risk-averse settings, though, semideviation and first-order stochastic dominance can be appealing alternatives to provide significantly more stable production planning costs with a marginal increase of the expected costs.     

Preliminary Evidence on the Diagnostic and Molecular Role of Circulating Soluble EGFR in Non-Small Cell Lung Cancer

Assessment of biological diagnostic factors providing clinically-relevant information to guide physician decision-making are still needed for diseases with poor outcomes, such as non-small cell lung cancer (NSCLC). Epidermal growth factor receptor (EGFR) is a promising molecule in the clinical management of NSCLC. While the EGFR transmembrane form has been extensively investigated in large clinical trials, the soluble, circulating EGFR isoform (sEGFR), which may have a potential clinical use, has rarely been considered. This study investigates the use of sEGFR as a potential diagnostic biomarker for NSCLC and also characterizes the biological function of sEGFR to clarify the molecular mechanisms involved in the course of action of this protein. Plasma sEGFR levels from a heterogeneous cohort of 37 non-advanced NSCLC patients and 54 healthy subjects were analyzed by using an enzyme-linked immunosorbent assay. The biological function of sEGFR was analyzed in vitro using NSCLC cell lines, investigating effects on cell proliferation and migration. We found that plasma sEGFR was significantly decreased in the NSCLC patient group as compared to the control group (median value: 48.6 vs. 55.6 ng/mL respectively; p = 0.0002). Moreover, we demonstrated that sEGFR inhibits growth and migration of NSCLC cells in vitro through molecular mechanisms that included perturbation of EGF/EGFR cell signaling and holoreceptor internalization. These data show that sEGFR is a potential circulating biomarker with a physiological protective role, providing a first approach to the functional role of the soluble isoform of EGFR. However, the impact of these data on daily clinical practice needs to be further investigated in larger prospective studies.     

Pseudouridine synthase 7 impacts Candida albicans rRNA processing and morphological plasticity

RNA can be modified in over 100 distinct ways, and these modifications are critical for function. Pseudouridine synthases catalyse pseudouridylation, one of the most prevalent RNA modifications. Pseudouridine synthase 7 modifies a variety of substrates in Saccharomyces cerevisiae including tRNA, rRNA, snRNA, and mRNA, but the substrates for other budding yeast Pus7 homologues are not known. We used CRISPR-mediated genome editing to disrupt Candida albicans PUS7 and find absence leads to defects in rRNA processing and a decrease in cell surface hydrophobicity. Furthermore, C. albicans Pus7 absence causes temperature sensitivity, defects in filamentation, altered sensitivity to antifungal drugs, and decreased virulence in a wax moth model. In addition, we find C. albicans Pus7 modifies tRNA residues, but does not modify a number of other S. cerevisiae Pus7 substrates. Our data suggests C. albicans Pus7 is important for fungal vigour and may play distinct biological roles than those ascribed to S. cerevisiae Pus7.     

Sustainable Production of Surfactants from Renewable Resources(Online First, Recommended Article)

Surfactants are important chemical products, serving as emulsifiers and interfacial modifiers in the household detergents, personal care products, paints and coatings, foods, cosmetics, and pharmaceuticals industries. This review focuses upon recent advances in research and development to improve the ecological sustainability of surfactants throughout their life cycle, including derivation from renewable resources, production using green manufacturing principles, and improved biocompatibility and biodegradability during their consumer use and disposal stages. Biobased surfactants, derived from vegetable oils, polysaccharides, proteins, phospholipids, and other renewable resources, currently comprise approximately 24% of the surfactant market, and this percentage is expected to increase, especially in Asia. The use of renewables is attractive to consumers because of reduced production of CO2, a greenhouse gas associated with climate change. Enzymes can greatly increase process sustainability, through reduced use of organic solvent, water, and energy, and reduced formation of by-products and waste products. Among the enzymes being investigated for surfactant synthesis, lipases are the most robust, due to their relatively high biocatalytic activity, operational stability and their ability to form or cleave ester, amide, and thioester bonds. For enzymes to be robust catalysts of surfactants, further research and development is needed to improve catalytic productivity, stability and reduce their purchase cost.     

Cyp1 Inhibition Prevents Doxorubicin-Induced Cardiomyopathy in a Zebrafish Heart-Failure Model

Doxorubicin is a highly effective chemotherapy agent used to treat many common malignancies. However, its use is limited by cardiotoxicity, and cumulative doses exponentially increase the risk of heart failure. To identify novel heart failure treatment targets, a zebrafish model of doxorubicin-induced cardiomyopathy was previously established for small-molecule screening. Using this model, several small molecules that prevent doxorubicin-induced cardiotoxicity both in zebrafish and in mouse models have previously been identified. In this study, exploration of doxorubicin cardiotoxicity is expanded by screening 2271 small molecules from a proprietary, target-annotated tool compound collection. It is found that 120 small molecules can prevent doxorubicin-induced cardiotoxicity, including 7 highly effective compounds. Of these, all seven exhibited inhibitory activity towards cytochrome P450 family 1 (CYP1). These results are consistent with previous findings, in which visnagin, a CYP1 inhibitor, also prevents doxorubicin-induced cardiotoxicity. Importantly, genetic mutation of cyp1a protected zebrafish against doxorubicin-induced cardiotoxicity phenotypes. Together, these results provide strong evidence that CYP1 is an important contributor to doxorubicin-induced cardiotoxicity and highlight the CYP1 pathway as a candidate therapeutic target for clinical cardioprotection.