The toxbox: specific DNA sequence requirements for activation of Vibrio cholerae virulence genes by ToxT

The Gram-negative, curved rod Vibrio cholerae causes the severe diarrhoeal disease cholera. The two major virulence factors produced by V. cholerae during infection are the cholera toxin (CT) and the toxin-coregulated pilus (TCP). Transcription of the genes encoding both CT and the components of the TCP is directly activated by ToxT, a transcription factor in the AraC/XylS family. ToxT binds upstream of the ctxAB genes, encoding CT, and upstream of tcpA , the first gene in a large operon encoding the components of the TCP. The DNA sequences upstream of ctxAB and tcpA that contain ToxT binding sites do not have any significant similarity other than being AT-rich. Extensive site-directed mutagenesis was performed on the region upstream of tcpA previously shown to be protected by ToxT, and we identified specific base pairs important for activation of tcpA transcription by ToxT. This genetic approach was complemented by copper-phenanthroline footprinting experiments that showed protection by ToxT of the base pairs identified as most important for transcription activation in the mutagenesis experiments. Based on this new information and on previous work, we propose the presence of a ToxT-binding motif – the ‘toxbox’– in promoters regulated by ToxT. At tcpA , two toxbox elements are present in a direct repeat configuration and both are required for activation of transcription by ToxT. The identity of only a few of the base pairs within the toxbox is important for activation by ToxT, and we term these the core toxbox elements. Lastly, we examined ToxT binding to a mutant having 5 bp inserted between the two toxboxes at tcpA and found that occupancy of both binding sites is retained regardless of the positions of the binding sites relative to each other on the face of the DNA. This suggests that ToxT binds independently as a monomer to each toxbox in the tcpA direct repeat, in accordance with what we observed previously with the inverted repeat ToxT sites between acfA and acfD .Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/75755/1/j.1365-2958.2006.05053.x.pd

Clinical evaluation of two dark blood methods of late gadolinium quantification of ischemic scar

BACKGROUND: Late gadolinium enhancement (LGE) imaging was validated for diagnosis and quantification of myocardial infarction (MI). Despite good contrast between scar and normal myocardium, contrast between blood pool and myocardial scar can be limited. Dark blood LGE sequences attempt to overcome this issue. PURPOSE: To evaluate T1 rho (T1 ρ)-prepared dark blood sequence and compare to blood nulled (BN) phase sensitive inversion recovery (PSIR) and standard myocardium nulled (MN) PSIR for detection and quantification of scar. STUDY TYPE: Prospective. POPULATION: Thirty patients with prior MI. FIELD STRENGTH/SEQUENCE: Patients underwent identical 1.5 T MRI protocols. Following routine LGE imaging, a slice with scar, remote myocardium, and blood pool was selected. PSIR LGE was repeated with inversion time set to MN, to BN, and T1 ρ FIDDLE (flow-independent dark-blood delayed enhancement) in random order. ASSESSMENT: Three observers. Qualitative assessment of confidence scores in scar detection and degree of transmurality. Quantitative assessment of myocardial scar mass (grams), and contrast-to-noise ratio (CNR) measurements between scar, blood pool, and myocardium. STATISTICAL TESTS: Repeated-measures analysis of variance (ANOVA) with Bonferroni correction, coefficient of variation, and the Cohen κ statistic. RESULTS: CNRscar-blood was significantly increased for both BN (27.1 ± 10.4) and T1 ρ (30.2 ± 15.1) compared with MN (15.3 ± 8.4 P < 0.001 for both sequences). There was no significant difference in CNRscar-myo between BN (55.9 ± 17.3) and MN (51.1 ± 17.8 P = 0.512); both had significantly higher CNRscar-myo compared with the T1 ρ (42.6 ± 16.9 P = 0.007 and P = 0.014, respectively). No significant difference in scar size between LGE methods: MN (2.28 ± 1.58 g) BN (2.16 ± 1.57 g) and T1 ρ (2.29 ± 2.5 g). Confidence scores were significantly higher for BN (3.87 ± 0.346) compared with MN (3.1 ± 0.76 P < 0.001) and T1 ρ (3.20 ± 0.71 P < 0.001). DATA CONCLUSION: PSIR with inversion time (TI) set for blood nulling and the T1 ρ LGE sequence demonstrated significantly higher scar to blood CNR compared with routine MN. PSIR with TI set for blood nulling demonstrated significantly higher reader confidence scores compared with routine MN and T1 ρ LGE, suggesting routine adoption of a BN PSIR approach might be appropriate for LGE imaging. LEVEL OF EVIDENCE: 2 Technical Efficacy: Stage 2 J. Magn. Reson. Imaging 2018

Impacts of climate change on national biodiversity population trends

Climate change has had well-documented impacts on the distribution and phenology of species across many taxa, but impacts on species’ abundance, which relates closely to extinction risk and ecosystem function, have not been assessed across taxa. In the most comprehensive multi-taxa comparison to date, we modelled variation in national population indices of 501 mammal, bird, aphid, butterfly and moth species as a function of annual variation in weather variables, which through time allowed us to identify a component of species’ population growth that can be associated with post-1970s climate trends. We found evidence that these climate trends have significantly affected population trends of 15.8% of species, including eight with extreme (> 30% decline per decade) negative trends consistent with detrimental impacts of climate change. The modelled effect of climate change could explain 48% of the significant across-species population decline in moths and 63% of the population increase in winged aphids. The other taxa did not have significant across-species population trends or consistent climate change responses. Population declines in species of conservation concern were linked to both climatic and non-climatic factors respectively accounting for 42 and 58% of the decline. Evident differential impacts of climate change between trophic levels may signal the potential for future ecosystem disruption. Climate change has therefore already driven large-scale population changes of some species, had significant impacts on the overall abundance of some key invertebrate groups and may already have altered biological communities and ecosystems in Great Britain

A new approach to modelling the relationship between annual population abundance indices and weather data

Weather has often been associated with fluctuations in population sizes of species; however, it can be difficult to estimate the effects satisfactorily because population size is naturally measured by annual abundance indices whilst weather varies on much shorter timescales. We describe a novel method for estimating the effects of a temporal sequence of a weather variable (such as mean temperatures from successive months) on annual species abundance indices. The model we use has a separate regression coefficient for each covariate in the temporal sequence, and over-fitting is avoided by constraining the regression coefficients to lie on a curve defined by a small number of parameters. The constrained curve is the product of a periodic function, reflecting assumptions that associations with weather will vary smoothly throughout the year and tend to be repetitive across years, and an exponentially decaying term, reflecting an assumption that the weather from the most recent year will tend to have the greatest effect on the current population and that the effect of weather in previous years tends to diminish as the time lag increases. We have used this approach to model 501 species abundance indices from Great Britain and present detailed results for two contrasting species alongside an overall impression of the results across all species. We believe this approach provides an important advance to the challenge of robustly modelling relationships between weather and species population size

Professionalism, Golf Coaching and a Master of Science Degree: A commentary

As a point of reference I congratulate Simon Jenkins on tackling the issue of professionalism in coaching. As he points out coaching is not a profession, but this does not mean that coaching would not benefit from going through a professionalization process. As things stand I find that the stimulus article unpacks some critically important issues of professionalism, broadly within the context of golf coaching. However, I am not sure enough is made of understanding what professional (golf) coaching actually is nor how the development of a professional golf coach can be facilitated by a Master of Science Degree (M.Sc.). I will focus my commentary on these two issues

Solar flare prediction using advanced feature extraction, machine learning and feature selection

YesNovel machine-learning and feature-selection algorithms have been developed to study: (i) the flare prediction capability of magnetic feature (MF) properties generated by the recently developed Solar Monitor Active Region Tracker (SMART); (ii) SMART's MF properties that are most significantly related to flare occurrence. Spatio-temporal association algorithms are developed to associate MFs with flares from April 1996 to December 2010 in order to differentiate flaring and non-flaring MFs and enable the application of machine learning and feature selection algorithms. A machine-learning algorithm is applied to the associated datasets to determine the flare prediction capability of all 21 SMART MF properties. The prediction performance is assessed using standard forecast verification measures and compared with the prediction measures of one of the industry's standard technologies for flare prediction that is also based on machine learning - Automated Solar Activity Prediction (ASAP). The comparison shows that the combination of SMART MFs with machine learning has the potential to achieve more accurate flare prediction than ASAP. Feature selection algorithms are then applied to determine the MF properties that are most related to flare occurrence. It is found that a reduced set of 6 MF properties can achieve a similar degree of prediction accuracy as the full set of 21 SMART MF properties

Scale-free static and dynamical correlations in melts of monodisperse and Flory-distributed homopolymers: A review of recent bond-fluctuation model studies

It has been assumed until very recently that all long-range correlations are screened in three-dimensional melts of linear homopolymers on distances beyond the correlation length $\xi$ characterizing the decay of the density fluctuations. Summarizing simulation results obtained by means of a variant of the bond-fluctuation model with finite monomer excluded volume interactions and topology violating local and global Monte Carlo moves, we show that due to an interplay of the chain connectivity and the incompressibility constraint, both static and dynamical correlations arise on distances $r \gg \xi$. These correlations are scale-free and, surprisingly, do not depend explicitly on the compressibility of the solution. Both monodisperse and (essentially) Flory-distributed equilibrium polymers are considered.Comment: 60 pages, 49 figure