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Showing how biophysics and biomedical engineering have advanced modern medicine, this text provides a multidisciplinary understanding of biological phenomena and the instrumentation for monitoring these phenomena. It also explores the application of physics and engineering methods to medicine and biology. The book includes a range of numerical problems with worked solutions and offers MATLAB code for advanced mathematical analysis of physiological and clinical monitoring systems. A solutions manual is available with qualifying course adoption.

QUESTION: Is adjusting the choke a normal maintenance step when a mechanic does a tune-up? I had a tune-up done recently and it seems that the choke is no better than when I brought the car in. You would think that the mechanic could have done the choke; he...

QUESTION: The other day on the freeway one of those gravel trucks lost a rock from the back of the truck which hit the windshield of my car leaving a small hole. The truck's insurance company said that they would be willing to repair the windshield for me, but...

Oil Change: Changing your engine oil is simple and since this is probably the most frequent regular maintenance item, the \"do it yourselfer\" can save a lot of money.

QUESTION: The other day on the freeway one of those gravel trucks lost a rock from the back of the truck which hit the windshield of my car leaving a small hole. The truck's insurance company said that they would be willing to repair the windshield for me, but...

Crystallization of RNA molecules can pose a challenge, and the U1A RNA binding protein has been used to facilitate the process. Now a different portable RNA sequence and a synthetic Fab are presented as tools for RNA structural determination. In addition to functioning as a crystallization chaperone, the Fab also serves as the search model to provide phase information. RNA crystallization and phasing represent major bottlenecks in RNA structure determination. Seeking to exploit antibody fragments as RNA crystallization chaperones, we have used an arginine-enriched synthetic Fab library displayed on phage to obtain Fabs against the class I ligase ribozyme. We solved the structure of a Fab–ligase complex at 3.1-Å resolution using molecular replacement with Fab coordinates, confirming the ribozyme architecture and revealing the chaperone's role in RNA recognition and crystal contacts. The epitope resides in the GAAACAC sequence that caps the P5 helix, and this sequence retains high-affinity Fab binding within the context of other structured RNAs. This portable epitope provides a new RNA crystallization chaperone system that easily can be screened in parallel to the U1A RNA-binding protein, with the advantages of a smaller loop and Fabs′ high molecular weight, large surface area and phasing power.

Despite its relative infancy, ecological science plays a pre‐eminent role in current environmental decision‐making globally and has, over recent decades, permeated a broad range of academic disciplines. Developments in two areas of philosophical thought in particular, environmental aesthetics and the aesthetics of science, beg an exploration of their intersection with respect to the role of aesthetics in ecological science. Here, we provide a contemporary synthesis of both environmental aesthetics and aesthetics of science to explore aesthetic dimensions of contemporary ecological science, highlighting three main areas of convergence: (1) the influence of aesthetic experiences and judgements of nature by ecologists on ecological science and our contemporary understanding of nature; (2) the development and role of ecological ‘taste’ among ecologists; and (3) moral, cultural and political implications of the ecological imagination as underpinned by current ecological science. We identify a risk for feedback mechanisms to perpetuate a relatively homogeneous ecological aesthetic as a result of reciprocal influences between ecological science and society which may further promote inadvertent policy advocacy and stifle scientific innovation. We suggest ecological science would benefit from increased aesthetic literacy and reflection by broadening the ecological imagination and intentionally facilitating more diverse and equitable science to inform policy outcomes. Our argument should be of interest to philosophers of science, ecologists and those that draw on their outputs. Read the free Plain Language Summary for this article on the Journal blog. Read the free Plain Language Summary for this article on the Journal blog.

The ability to genetically manipulate microorganisms has been essential for understanding their biology and metabolism. Targeted genome editing relies on highly efficient homologous recombination, and while this is readily observed in the yeast Saccharomyces cerevisiae, most non-conventional yeast species do not display this trait and remain recalcitrant to targeted editing methods. CRISPR-based editing can bypass the requirement for high levels of native homologous recombination, enabling targeted modification to be more broadly implemented. While genetic transformation has been reported previously in Brettanomyces bruxellensis, a yeast with broad biotechnological potential and responsible for significant economic losses during the production of fermented beverages, targeted editing approaches have not been reported. Here, we describe the use of an expression-free CRISPR-Cas9 system, in combination with gene transformation cassettes tailored for B. bruxellensis, to provide the means for targeted gene deletion in this species. Deletion efficiency was shown to be dependent on homologous flanking DNA length, with higher targeting efficiencies observed with cassettes containing longer flanking regions. In a diploid strain, it was not possible to delete multiple alleles in one step, with heterozygous deletants only obtained when using DNA cassettes with long flanking regions. However, stepwise transformations (using two different marker genes) were successfully used to delete both wild-type alleles. Thus, the approach reported here will be crucial to understand the complex physiology of B. bruxellensis.Key points• The use of CRISPR-Cas9 enables targeted gene deletion in Brettanomyces bruxellensis.• Homozygous diploid deletions are possible with step-wise transformations.• Deletion of SSU1 confirmed the role of this gene in sulphite tolerance.

ABSTRACT Brettanomyces bruxellensis is considered one of the most problematic microbes associated with wine production. Sulfur dioxide is commonly used to inhibit the growth of B. bruxellensis and limit the potential wine spoilage. Brettanomyces bruxellensis wine isolates can grow at higher concentrations of this preservative than isolates from other sources. Thus, it has been suggested that the use of sulfite may have selected for B. bruxellensis strains better adapted to survive in the winemaking environment. We utilized laboratory adaptive evolution to determine the potential for this to occur. Three B. bruxellensis strains, representative of known genetic variation within the species, were subjected to increasing sublethal sulfur dioxide concentrations. Individual clones isolated from evolved populations displayed enhanced sulfite tolerance, ranging from 1.6 to 2.5 times higher than the corresponding parental strains. Whole-genome sequencing of sulfite-tolerant clones derived from two of the parental strains revealed structural variations affecting 270 genes. The region containing the sulfite efflux pump encoding gene, SSU1, showed clear copy number variants in all sequenced clones. Regardless of parental strain genetic background, SSU1 copy number changes were reproducibly associated with one SSU1 haplotype. This work clearly demonstrates adaptive evolution of B. bruxellensis when exposed to sublethal sulfites and suggests that, similar to Saccharomyces cerevisiae wine yeast, the mechanism responsible involves the gene SSU1. The wine spoilage yeast Brettanomyces bruxellensis is usually inhibited by sulfur dioxide during winemaking; however, this yeast has the potential to develop increased tolerance to this preservative.

In most yeast-driven biotechnological applications, biomass is separated from the aqueous phase after fermentation or production has finished. During winemaking, yeasts are removed after fermentation by racking, filtration, or centrifugation, which add costs to the overall process and may reduce product yield. Theoretically, clarification and filtration can be aided through use of yeast strains that form flocs due to cell-cell binding, a process known as flocculation. However, because early flocculation can cause stuck/sluggish fermentations, this phenotype is not common amongst commercially available wine yeasts. In this study we sought to identify wine strains that exhibit late-fermentation flocculant behaviour using two complementary approaches; a high-throughput sedimentation rate assay of individual strains and a competitive sedimentation assay using a barcoded yeast collection. Amongst 103 wine strains, several exhibited strong sedimentation at the end of the wine fermentation process under various environmental conditions. Two of these strains, AWRI1688 and AWRI1759, were further characterised during red winemaking trials. Shiraz wines produced with both strains displayed improved filtration-related properties. AWRI1759 produced wines with greater filterability, whereas AWRI1688 enabled the recovery of larger wine volumes after racking. Thus, this study demonstrates the effective use of sedimentation screening assays to identify wine yeasts with practical winemaking applications.