Expanding the Genetic Code of and to Incorporate Non-canonical Amino Acids for Production of Modified Lantibiotics

The incorporation of non-canonical amino acids (ncAAs) into ribosomally synthesized and post-translationally modified peptides, e.g., nisin from the Gram-positive bacterium Lactococcus lactis, bears great potential to expand the chemical space of various antimicrobials. The ncAA Nε-Boc-L-lysine (BocK) was chosen for incorporation into nisin using the archaeal pyrrolysyl-tRNA synthetase–tRNAPyl pair to establish orthogonal translation in L. lactis for read-through of in-frame amber stop codons. In parallel, recombinant nisin production and orthogonal translation were combined in Escherichia coli cells. Both organisms synthesized bioactive nisin(BocK) variants. Screening of a nisin amber codon library revealed suitable sites for ncAA incorporation and two variants displayed high antimicrobial activity. Orthogonal translation in E. coli and L. lactis presents a promising tool to create new-to-nature nisin derivatives

A novel class of heat and secretion stress-responsive genes is controlled by the autoregulated CssRS two-component system of Bacillus subtilis

Bacteria need dedicated systems that allow appropriate adaptation to the perpetual changes in their environments. In Bacillus subtilis, two HtrA-like proteases, HtrA and HtrB, play critical roles in the cellular response to secretion and heat stresses. Transcription of these genes is induced by the high-level production of a secreted protein or by a temperature upshift. The CssR-CssS two-component regulatory system plays an essential role in this transcriptional activation. Transcription of the cssRS operon is autoregulated and can be induced by secretion stress, by the absence of either HtrA or HtrB, and by heat stress in a HtrA null mutant strain. Two start sites are used for cssRS transcription, only one of which is responsive to heat and secretion stress. The divergently transcribed htrB and cssRS genes share a regulatory region through which their secretion and heat stress-induced expression is linked. This study shows that CssRS-regulated genes represent a novel class of heat-inducible genes, which is referred to as class V and currently includes two genes: htrA and htrB

Ergodicity of certain cocycles over certain interval exchanges

We show that for odd-valued piecewise-constant skew products over a certain two parameter family of interval exchanges, the skew product is ergodic for a full-measure choice of parameters

Neural processing of criticism and positive comments from relatives in individuals with schizotypal personality traits

Objectives. High negative expressed emotion by family members towards schizophrenia patients increases the risk of subsequent relapse. The study aimed to determine whether individuals with high schizotypy (HS) and low schizotypy (LS) would differ in activation of brain areas involved in cognitive control when listening to relative criticism

Centuries of intense surface melt on Larsen C Ice Shelf

Following a southward progression of ice-shelf disintegration along the Antarctic Peninsula, Larsen C Ice Shelf is the focus of ongoing investigation regarding its future stability. The ice shelf is known to be experience surface melt, and commonly features surface meltwater ponds. Here, we use a flowline model and a firn density model to date and interpret observations of melt-affected ice layers found within five 90?m boreholes distributed across the ice shelf. We find that units of ice within the boreholes, which have densities exceeding those expected under normal compaction metamorphism, correspond to two climatic warm periods within the last 300 years on the Antarctic Peninsula. The more recent warm period, from the 1960s onwards, has generated distinct sections of dense ice in two boreholes in Cabinet Inlet, close to the Antarctic Peninsula mountains ? a region currently affected by f?hn winds. Previous work has classified these layers as refrozen pond ice, requiring large quantities of mobile liquid water to form. Our flowline model shows that, whilst preconditioning of the ice began in the late 1960s, it was probably not until the early 1990s that twentieth-century ponding began. The earlier warm period occurred during the 18th century and resulted in two additional sections of anomalously dense ice deep within the boreholes. The first, in one of the Cabinet Inlet boreholes, consists of ice characteristic of refrozen ponds and must have formed in an area currently featuring ponding. The second, in a mid-shelf borehole, formed at the same time in an area which now experiences significant annual melt. Further south on the shelf, the boreholes sample ice that is of an equivalent age but which does not exhibit the same degree of melt influence. This west?east and north?south gradient in past melt distribution resembles current spatial patterns of surface melt intensity. Using flowlines to trace the advection and submergence of continental ice identified in boreholes, we demonstrate that, even by the time the ice reaches the calving front, only the upper 40 to 50?% of the shelf is composed of meteoric ice accumulated on the shelf. This vertical composition implies that basal crevasses must be confined within continental and/or basally accreted ice, and therefore will be unaffected by current climate-induced firn compactionauthorsversio

Ice and firn heterogeneity within Larsen C Ice Shelf from borehole optical televiewing

Research was funded by the UK Natural Environmental Research Council grants NE/L006707/1 and NE/L005409/1 and a HEFCW/Aberystwyth University Capital Equipment Grant to B.H. Data will be available via the project website (www.projectmidas.org) and the UK Polar Data Centre (https://www.bas.ac.uk/data/uk-pdc/) from mid-2017.We use borehole optical televiewing (OPTV) to explore the internal structure of Larsen C Ice Shelf (LCIS). We report a suite of five ~90 m long OPTV logs, recording a light-emitting diode-illuminated, geometrically correct image of the borehole wall, from the northern and central sectors of LCIS collected during austral spring 2014 and 2015. We use a thresholding-based technique to estimate the refrozen ice content of the ice column and exploit a recently calibrated density-luminosity relationship to reveal its structure. All sites are dense and strongly influenced by surface melt, with frequent refrozen ice layers and mean densities, between the depths of 1.87 and 90 m, ranging from 862 to 894 kg m−3. We define four distinct units that comprise LCIS and relate these to ice provenance, dynamic history, and past melt events. These units are in situ meteoric ice with infiltration ice (U1), meteoric ice which has undergone enhanced densification (U2), thick refrozen ice (U3), and advected continental ice (U4). We show that the OPTV-derived pattern of firn air content is consistent with previous estimates, but that a significant proportion of firn air is contained within U4, which we interpret to have been deposited inland of the grounding line. The structure of LCIS is strongly influenced by the E-W gradient in föhn-driven melting, with sites close to the Antarctic Peninsula being predominantly composed of refrozen ice. Melting is also substantial toward the ice shelf center with >40% of the overall imaged ice column being composed of refrozen ice.Publisher PDFPeer reviewe

Flow-line model code for accumulation of ice along velocity-based trajectories

The flow-line model was designed to enable estimation of the age and surface origin for various ice bodies identified within hot-water drilled boreholes on Larsen C Ice Shelf. Surface fluxes are accumulated, converted to thicknesses, and advected down flow from a fixed number of selected points. The model requires input datasets of surface mass balance, surface velocity, vertical strain rates, ice-shelf thickness, and a vertical density profile. This model is part of a larger project. Input datasets such as density profiles and trajectory vectors are available separately. Resolution is dependent on the input datasets. Funding was provided by the NERC grant NE/L005409/1

Massive subsurface ice formed by refreezing of ice-shelf melt ponds

Surface melt ponds form intermittently on several Antarctic ice shelves. Although implicated in ice-shelf break up, the consequences of such ponding for ice formation and ice-shelf structure have not been evaluated. Here we report the discovery of a massive subsurface ice layer, at least 16 km across, several kilometres long and tens of metres deep, located in an area of intense melting and intermittent ponding on Larsen C Ice Shelf, Antarctica. We combine borehole optical televiewer logging and radar measurements with remote sensing and firn modelling to investigate the layer, found to be ∼10 C warmer and ∼170 kg m¯³ denser than anticipated in the absence of ponding and hitherto used in models of ice-shelf fracture and flow. Surface ponding and ice layers such as the one we report are likely to form on a wider range of Antarctic ice shelves in response to climatic warming in forthcoming decades

Changes in cerebral oxygenation and cerebral blood flow during hemodialysis - A simultaneous near-infrared spectroscopy and positron emission tomography study

Near-infrared spectroscopy (NIRS) is used to monitor cerebral tissue oxygenation (rSO2) depending on cerebral blood flow (CBF), cerebral blood volume and blood oxygen content. We explored whether NIRS might be a more easy applicable proxy to [15O]H2O positron emission tomography (PET) for detecting CBF changes during hemodialysis. Furthermore, we compared potential determinants of rSO2 and CBF. In 12 patients aged ≥ 65 years, NIRS and PET were performed simultaneously: before (T1), early after start (T2), and at the end of hemodialysis (T3). Between T1 and T3, the relative change in frontal rSO2 (ΔrSO2) was -8 ± 9% ( P = 0.001) and -5 ± 11% ( P = 0.08), whereas the relative change in frontal gray matter CBF (ΔCBF) was -11 ± 18% ( P = 0.009) and -12 ± 16% ( P = 0.007) for the left and right hemisphere, respectively. ΔrSO2 and ΔCBF were weakly correlated for the left (ρ 0.31, P = 0.4), and moderately correlated for the right (ρ 0.69, P = 0.03) hemisphere. The Bland-Altman plot suggested underestimation of ΔCBF by NIRS. Divergent associations of pH, pCO2 and arterial oxygen content with rSO2 were found compared to corresponding associations with CBF. In conclusion, NIRS could be a proxy to PET to detect intradialytic CBF changes, although NIRS and PET capture different physiological parameters of the brain