ENANTIOSELECTIVE BACTERIAL BIOTRANSFORMATION ROUTES TO CIS-DIOL METABOLITES OF MONOSUBSTITUTED BENZENES, NAPHTHALENE AND BENZOCYCLOALKENES OF EITHER ABSOLUTE-CONFIGURATION

Enzyme-catalysed kinetic resolution and asymmetric dihydroxylation routes to enantiopure cis-diol metabolites of arenes and benzocycloalkenes of either absolute configuration have been developed using appropriate strains of the bacterium Pseudomonas putida.

Phosphoramidates of 2'-beta-D-arabinouridine (AraU) as phosphate prodrugs; design, synthesis, in vitro activity and metabolism

2'-Beta-D-arabinouridine (AraU), the uridine analogue of the anticancer agent AraC, was synthesized and evaluated for antiviral activity and cytotoxicity. In addition, a series of AraU monophosphate prodrugs in the form of triester phosphoramidates (ProTides) were also synthesized and tested against a range of viruses, leukaemia and solid tumour cell lines. Unfortunately, neither the parent compound (AraU) nor any of its ProTides showed antiviral activity, nor potent inhibitory activity against any of the cancer cell lines. Therefore, the metabolism of AraU phosphoramidates to release AraU monophosphate was investigated. The results showed carboxypeptidase Y, hog liver esterase and crude CEM tumor cell extracts to hydrolyse the ester motif of phosphoramidates with subsequent loss of the aryl group, while molecular modelling studies suggested that the AraU l-alanine aminoacyl phosphate derivative might not be a good substrate for the phosphoramidase enzyme Hint-1. These findings are in agreement with the observed disappearance of intact prodrug and concomitant appearance of the corresponding phosphoramidate intermediate derivative in CEM cell extracts without measurable formation of araU monophosphate. These findings may explain the poor antiviral/cytostatic potential of the prodrugs.

Structural and kinetic characterization of the LPS biosynthetic enzyme D-alpha,beta-D-heptose-1,7-bisphosphate phosphatase (GmhB) from Escherichia coli

Lipopolysaccharide is a major component of the outer membrane of gram-negative bacteria and provides a permeability barrier to many commonly used antibiotics. ADP-heptose residues are an integral part of the LPS inner core, and mutants deficient in heptose biosynthesis demonstrate increased membrane permeability. The heptose biosynthesis pathway involves phosphorylation and dephosphorylation steps not found in other pathways for the synthesis of nucleotide sugar precursors. Consequently, the heptose biosynthetic pathway has been marked as a novel target for antibiotic adjuvants, which are compounds that facilitate and potentiate antibiotic activity. D-alpha,beta-D-heptose-1,7-bisphosphate phosphatase (GmhB) catalyzes the third essential step of LPS heptose biosynthesis. This study describes the first crystal structure of GmhB and enzymatic analysis of the protein. Structure-guided mutations followed by steady state kinetic analysis, together with established precedent for HAD phosphatases, suggest that GmhB functions through a phosphoaspartate intermediate. This study provides insight into the structure-function relationship of GmhB, a new target for combatting gram-negative bacterial infection.

Skin Dendritic Cell Targeting via Microneedle Arrays Laden with Antigen-Encapsulated Poly-D, L-lactide-co-Glycolide Nanoparticles Induces Efficient Antitumor and Antiviral Immune Responses

The efficacious delivery of antigens to antigen-presenting cells (APCs), in particular, to dendritic cells (DCs), and their subsequent activation remains a significant challenge in the development of effective vaccines. This study highlights the potential of dissolving microneedle (MN) arrays laden with nanoencapsulated antigen to increase vaccine immunogenicity by targeting antigen specifically to contiguous DC networks within the skin. Following in situ uptake, skin-resident DCs were able to deliver antigen-encapsulated poly-d,l-lactide-co-glycolide (PGLA) nanoparticles to cutaneous draining lymph nodes where they subsequently induced significant expansion of antigen-specific T cells. Moreover, we show that antigen-encapsulated nanoparticle vaccination via microneedles generated robust antigen-specific cellular immune responses in mice. This approach provided complete protection in vivo against both the development of antigen-expressing B16 melanoma tumors and a murine model of para-influenza, through the activation of antigen-specific cytotoxic CD8(+) T cells that resulted in efficient clearance of tumors and virus, respectively. In addition, we show promising findings that nanoencapsulation facilitates antigen retention into skin layers and provides antigen stability in microneedles. Therefore, the use of biodegradable polymeric nanoparticles for selective targeting of antigen to skin DC subsets through dissolvable MNs provides a promising technology for improved vaccination efficacy, compliance, and coverage.

Rayleigh-Taylor Instability of an Ultrathin Foil Accelerated by the Radiation Pressure of an Intense Laser

We report experimental evidence for a Rayleigh-Taylor-like instability driven by radiation pressure of an ultraintense (1021W/cm2) laser pulse. The instability is witnessed by the highly modulated profile of the accelerated proton beam produced when the laser irradiates a 5 nm diamondlike carbon (90% C, 10% H) target. Clear anticorrelation between bubblelike modulations of the proton beam and transmitted laser profile further demonstrate the role of the radiation pressure in modulating the foil. Measurements of the modulation wavelength, and of the acceleration from Doppler-broadening of back-reflected light, agree quantitatively with particle-in-cell simulations performed for our experimental parameters and which confirm the existence of this instability. © 2012 American Physical Society.

Sher 25: pulsating but apparently alone

The blue supergiant Sher 25 is surrounded by an asymmetric, hourglass-shaped circumstellar nebula, which shows similarities to the triple-ring structure seen around SN 1987A. From optical spectroscopy over six consecutive nights, we detect periodic radial velocity variations in the stellar spectrum of Sher 25 with a peak-to-peak amplitude of ~ 12 km s^-1 on a time-scale of about 6 d, confirming the tentative detection of similar variations by Hendry et al. From consideration of the amplitude and time-scale of the signal, coupled with observed line profile variations, we propose that the physical origin of these variations is related to pulsations in the stellar atmosphere, rejecting the previous hypothesis of a massive, short-period binary companion. The radial velocities of two other blue supergiants with similar bipolar nebulae, SBW1 and HD 168625, were also monitored over the course of six nights, but these did not display any significant radial velocity variations. 

Thiol isomerases negatively regulate the cellular shedding activity of ADAM17

ADAM17 (where ADAM is 'a disintegrin and metalloproteinase') can rapidly modulate cell-surface signalling events by the proteolytic release of soluble forms of proligands for cellular receptors. Many regulatory pathways affect the ADAM17 sheddase activity, but the mechanisms for the activation are still not clear. We have utilized a cell-based ADAM17 assay to show that thiol isomerases, specifically PDI (protein disulfide isomerase), could be responsible for maintaining ADAM17 in an inactive form. Down-regulation of thiol isomerases, by changes in the redox environment (for instance as elicited by phorbol ester modulation of mitochondrial reactive oxygen species) markedly enhanced ADAM17 activation. On the basis of ELISA binding studies with novel fragment antibodies against ADAM17 we propose that isomerization of the disulfide bonds in ADAM17, and the subsequent conformational changes, form the basis for the modulation of ADAM17 activity. The shuffling of disulfide bond patterns in ADAMs has been suggested by a number of recent adamalysin crystal structures, with distinct disulfide bond patterns altering the relative orientations of the domains. Such a mechanism is rapid and reversible, and the role of thiol isomerases should be investigated further as a potential factor in the redox regulation of ADAM17.

Biomineralization of Weathered Rock Rinds:Examples from the Lower Afroalpine Zone on Mount Kenya

Rock rinds have been used for half a century to date glacial deposits and recently inroads have been developed to use nuclides to provide absolute ages of weathering rinds in pebble clasts. Although maximum and minimum rind thicknesses have helped to elucidate time since deposition and allowed stratigraphic division of deposits at glacial rank, little has been done to investigate the wealth of mineral degradation, growth of alteration products and biomineralization that occur in these weathered crusts. In some cases the mass of microbe-mineral intergrowth is nearly present on a 50%/50% basis, with the biotic mass intergrown with mineral matter to such an extent that it probably controls pH and redox phenomena that act as accelerators in the weathering process. Assuming weathering time spans of 2 × 10⁶ years or more for a complete cycle, eventual clast decomposition is the end product. Here we present evidence of microbe-clast intergrowth from selected sites of Pleistocene age (~70 ka to 2.0 Ma) in the lower Afroalpine of Mt. Kenya and hypothesize about its role in rock decomposition and fossilization of biotic end-members. © 2013 Copyright Taylor and Francis Group, LLC.

Principles of fatigue in residency education: a qualitative study

Background: Proposals to implement fatigue-management strategies in residency education assume that medicine shares the view
of other risk-adverse industries that fatigue is hazardous. This view is an essential underpinning of fatigue-management strategies
that other industries have embedded as part of their workplace occupational health and safety programs. We sought to explore how
residents understand fatigue in the context of their training environment.

Methods: We interviewed 21 residents in 7 surgical and nonsurgical programs at Western University in 2014. All participants met the
inclusion criteria of routinely working 24-hour call shifts while enrolled in their training program. Data collection and analysis occurred iteratively in keeping with constructivist grounded theory methodology and informed theoretical sampling to sufficiency.

Results: Four predominant principles of fatigue captured how the social learning environment shaped residents’ perceptions of
fatigue. These included the conceptualization of fatigue as (a) inescapable and therefore accepted, (b) manageable through experience, (c) necessary for future practice and (d) surmountable when required.

Interpretation: This study elaborates our understanding of how principles of fatigue are constructed and reinforced by the training
environment. Whereas fatigue is seen as a collective hazard in other industries, our data showed that, in residency training, fatigue
may be seen as a personal challenge. Consequently, fatigue-management strategies that conceptualize fatigue as an occupational
threat may have a limited impact on resident behaviour and patient safety.

Address delivered by Chief Justice Eugene B. Gary, L.L.D. at the dedication of the Court House, McCormick County, September 27th, 1923

This document is an address by Chief Justice Eugene B. Gary that discusses the civilization of America in two contexts. The first context is the principles of civilization applicable to their democratic form of government. The second context is the dangers that threaten to destroy the very foundation of their government.