Genetic Encoding of para-Pentafluorosulfanyl Phenylalanine: A Highly Hydrophobic and Strongly Electronegative Group for Stable Protein Interactions.

SF5Phe, para-pentafluorosulfanyl phenylalanine, is an unnatural amino acid with extreme physicochemical properties, which is stable in physiological conditions. Here we present newly developed aminoacyl-tRNA synthetases that enable genetic encoding of SF5Phe for site-specific incorporation into proteins in high yields. Owing to the SF5 moiety's dichotomy of strong polarity and high hydrophobicity, the unnatural amino acid forms specific and strong interactions in proteins. The potential of SF5Phe in protein research is illustrated by (i) increasing the binding affinity of a consensus pentapeptide motif toward the ß subunit of Escherichia coli DNA polymerase III holoenzyme by mutation of a phenylalanine to a SF5Phe residue, (ii) site-specifically adhering ß-cyclodextrin to the surface of ubiquitin, and (iii) selective detection of 19F-19F nuclear Overhauser effects in the Escherichia coli peptidyl-prolyl cis/trans-isomerase B following mutation of two phenylalanine residues in the core of the protein to SF5Phe. With increasing use of the SF5 moiety in pharmaceutical chemistry, this general method of functionalizing proteins with SF5 groups opens unique opportunities for structural biology and in vivo studies.

Growth and Carcass Characteristics of Beef-Cross-Dairy-Breed Heifers and Steers Born to Different Dam Breeds.

Approximately two thirds of the annual beef kill in New Zealand originates from the dairy industry. The recent increase in Jersey genetics in the dairy herd will inevitably result in an increase in Jersey genetics entering the beef herd from retention of dairy-origin calves for finishing. Limited literature is available on the effect of dam breed on the performance of beef-cross-dairy-breed progeny. The aim of this study was to investigate the effect of dam breed from dams with varying proportions of Friesian and Jersey genetics on growth traits and carcass characteristics of their 24-month-old beef-cross-dairy-breed heifer and steer progeny. Liveweights of 142 heifers and 203 steers from Friesian (F), Friesian-cross (FX), Friesian-Jersey (FJ) and Jersey-cross (JX) dams were recorded at birth, weaning, as yearlings and at slaughter. Carcass characteristics were also recorded. At each point measured, liveweight was greatest for calves born to F dams. Calves born to F dams took 93 days to reach a weaning weight of 100 kg, whereas those from FX, FJ and JX dams took 99, 101 and 102 days, respectively. Carcass weight was greatest for progeny of F dams (286 kg, compared with 279, 275 and 276 for progeny of FX, FJ and JX dams, respectively). The progeny of JX dams had yellower fat than all other dam breed groups and a greater incidence of excessively yellow fat (fat score ≥ 5).

Turnover of quorum sensing signal molecules modulates cross-kingdom signalling.

N-acylhomoserine lactone (AHL) quorum-sensing molecules modulate the swimming behaviour of zoospores of the macroalga Ulva to facilitate the location of bacterial biofilms. Here we show that the intertidal surfaces colonized by Ulva are dominated by Alphaproteobacteria, particularly the Rhodobacteraceae family, and the Bacteroidetes family Flavobacteriaceae, and that this diverse assemblage both produces and degrades AHLs. N-acylhomoserine lactones could also be extracted from the surfaces of pebbles recovered from intertidal rock-pools. Bacteria representative of this assemblage were isolated and tested for the production and degradation of AHLs, and for their ability to modulate zoospore settlement at different biofilm densities. Of particular interest was a Shewanella sp. This strain produced three major AHLs (OC4, OC10 and OC12) in the late exponential phase, but the longer-chain AHLs were rapidly degraded in the stationary phase. Degradation occurred via both lactonase and amidase activity. A close relationship was found between AHL synthesis and Ulva zoospore settlement. The Shewanella isolate also interfered with AHL production by a Sulfitobacter isolate and its ability to enhance zoospore settlement in a polymicrobial biofilm. This influence on the attachment of Ulva zoospores suggests that AHL-degrading strains can affect bacterial community behaviour by interfering with quorum sensing between neighbouring bacteria. More importantly, these interactions may exert wider ecological effects across different kingdoms.

'They've just got symptoms without science': Medical trainees' acquisition of negative attitudes towards patients with medically unexplained symptoms.

OBJECTIVE: Doctors find patients with medically unexplained symptoms (MUS) challenging to manage and some hold negative attitudes towards these patients. It is unknown when and how these views form. This study examines medical trainees' beliefs and influences about MUS. METHODS: Semi-structured interviews with 43 medical trainees. Using an iterative approach, initial emergent themes were explored in subsequent interviews. Data generation continued until thematic saturation was achieved. RESULTS: Participants had received no training in MUS but had developed views about causes and management. They struggled with the concept of 'diagnosis by exclusion'. Attitudes towards patients had developed through informal clinical observation and interactions with doctors. Many welcomed formal training but identified a need to integrate theoretical learning with clinical application. CONCLUSION: Despite limited teaching, medical trainees are aware of the challenges in diagnosing and managing patients with MUS, acquiring attitudes through a hidden curriculum. To be welcomed, training must be evidence-based, theoretically informed, but clinically applicable. PRACTICAL IMPLICATIONS: Current medical training fails to equip doctors to engage with MUS and potentially fosters the development of unhelpful views of these patients. Informed teaching on diagnosis and management of MUS is necessary at a trainee level to limit the development of negative attitudes.