Identification of synergists that potentiate the action of polymyxin B against Burkholderia cenocepacia

Burkholderia cenocepacia and other members of the Burkholderia cepacia complex (Bcc) are highly multidrug-resistant bacteria that cause severe pulmonary infections in patients with cystic fibrosis. A screen of 2686 compounds derived from marine organisms identified molecules that could synergize with polymyxin B to inhibit growth of B. cenocepacia. At 1 μg/ml, five compounds synergized with polymyxin B and inhibited the growth of B. cenocepacia by more than 70% compared to growth in polymyxin B alone. Follow-up testing revealed that one compound from the screen, the aminocoumarin antibiotic novobiocin, synergized with polymyxin B and colistin against tobramycin-resistant clinical isolates of B. cenocepacia and Burkholderia multivorans. In parallel, we show that novobiocin sensitivity is common among Bcc species and these bacteria are even more susceptible to an alternative aminocoumarin, clorobiocin, which also had an additive effect with polymyxin B against B. cenocepacia. These studies support using aminocoumarin antibiotics to treat Bcc infections and show that synergizers can be found to increase the efficacy of antimicrobial peptides and polymyxins against Bcc bacteria.

Chemical compositions of four B-type supergiants in the SMC wing

High-resolution UCLES/AAT spectra of four B-type supergiants in the SMC South East Wing have been analysed using non-LTE model atmosphere techniques to determine their atmospheric parameters and chemical compositions. The principle aim of this analysis was to determine whether the very low metal abundances (-1.1 dex compared with Galactic value) previously found in the Magellanic Inter Cloud region (ICR) were also present in the SMC Wing. The chemical compositions of the four targets are similar to those found in other SMC objects and appear to be incompatible with those deduced previously for the ICR. Given the close proximity of the Wing to the ICR, this is difficult to understand and some possible explanations are briefly discussed.

The atmospheric parameters and chemical composition of early B-type giants in h and chi Persei

Atmospheric parameters and surface chemical compositions are presented for eight stars, classified as B1 or B2 but with a range of luminosity classes, in the northern double cluster h and chi Persei. Echelle spectroscopy (covering the wavelength region 3900 to 4700 Ä) and grating spectroscopy (of the Balmer, H? and Hß lines) were analysed using non-LTE synthetic spectra based on LTE line-blanketed atmosphere structures. High microturbulences are found in our sample, and this quantity must be included in the computation of the non-LTE level populations; its effect is generally to decrease the derived metal abundances by typically 0.1 dex but by up to 0.4 dex. Our absolute abundances are in reasonable agreement with those previously found for main sequence B-type stars, while we find some evidence for small abundance variations (particularly for nitrogen) within our sample. One star (BD+56 678) appears to be a spectrum variable and at two epochs shows a highly enriched nitrogen spectrum. Our atmospheric parameters imply that two stars have previously been mis-identified as main sequence objects and a distance modulus, at the higher end of the values previously deduced. The observational HR diagram is consistent with stellar evolutionary models that explicitly include the effects of rotation.

DPP IV resistance and insulin releasing activity of a novel di-substituted analogue of glucose-dependent insulinotropic polypeptide, (Ser(2)-Asp(13))GIP

Structure-function studies suggest that preservation of the N-terminus and secondary structure of glucose-dependent insulinotropic polypeptide (GIP) is important for biological activity. Therefore, a novel di-substituted analogue of GIP, (Ser(2)-Asp(13))GIP, containing a negatively charged Asp residue in place of an Ala in position 13, seas synthesised and evaluated for in vitro biological activity. Incubation with dipeptidyl peptidase IV (DPP IV) showed the half-lives of GIP and (Ser(2)-Asp(13))GIP to be 2.3 and >4 h, respectively. Insulin releasing studies in clonal pancreatic BRIN-BD11 cells demonstrated that (Ser(2)-Asp(13))GIP (10(-12) to 10(-7) mol/l) was significantly less potent (60-90%; P

Alpha-1-antitrypsin aerosolized augmentation abrogates neutrophil elastase-induced expression of cathepsin B and matrix metalloprotease 2 in vivo and in vitro

Background: Neutrophil elastase (NE) activity is increased in lung diseases such as a1-antitrypsin (A1AT) deficiency and pneumonia. It has recently been shown to induce expression of cathepsin B and matrix metalloprotease 2 (MMP-2) in vitro and in a mouse model. It is postulated that increased cathepsin B and MMP-2 in acute and chronic lung diseases result from high levels of extracellular NE and that expression of these proteases could be inhibited by A1AT augmentation therapy. <br/><br/>Methods: Cathepsin and MMP activities were assessed in bronchoalveolar lavage (BAL) fluid from patients with A1AT deficiency, pneumonia and control subjects. Macrophages were exposed to BAL fluid rich in free NE from patients with pneumonia following pretreatment with A1AT. MMP-2, cathepsin B, secretory leucoprotease inhibitor (SLPI) and lactoferrin levels were determined in BAL fluid from A1AT-deficient patients before and after aerosolisation of A1AT. <br/><br/>Results: BAL fluid from both patients with pneumonia and those with A1AT deficiency containing free NE had increased cathepsin B and MMP-2 activities compared with BAL fluid from healthy volunteers. The addition of A1AT to BAL fluid from patients with pneumonia greatly reduced NE-induced cathepsin B and MMP-2 expression in macrophages in vitro. A1AT augmentation therapy to A1AT-deficient individuals also reduced cathepsin B and MMP-2 activity in BAL fluid in vivo. Furthermore, A1AT-deficient patients had higher levels of SLPI and lactoferrin after A1AT augmentation therapy. <br/><br/>Conclusion: These findings suggest a novel role for A1AT inhibition of NE-induced upregulation of MMP and cathepsin expression both in vitro and in vivo.

Atmospheric parameters and rotational velocities for a sample of Galactic B-type supergiants

High-resolution optical spectra of 57 Galactic B-type supergiant stars have been analysed to determine their rotational and macroturbulent velocities. In addition, their atmospheric parameters (effective temperature, surface gravity and microturbulent velocity) and surface nitrogen abundances have been estimated using a non-local thermodynamic equilibrium grid of model atmospheres. Comparisons of the projected rotational velocities have been made with the predictions of stellar evolutionary models and in general good agreement was found. However, for a small number of targets, their observed rotational velocities were significantly larger than predicted, although their nitrogen abundances were consistent with the rest of the sample. We conclude that binarity may have played a role in generating their large rotational velocities. No correlation was found between nitrogen abundances and the current projected rotational velocities. However, a correlation was found with the inferred projected rotational velocities of the main-sequence precursors of our supergiant sample. This correlation is again in agreement with the predictions of single star evolutionary models that incorporate rotational mixing. The origin of the macroturbulence and microturbulent velocity fields is discussed and our results support previous theoretical studies that link the former to subphotospheric convection and the latter to non-radial gravity mode oscillations. In addition, we have attempted to identify differential rotation in our most rapidly rotating targets.

An ingress and a complete transit of HD80606 b

We have used four telescopes at different longitudes to obtain near-continuous light-curve coverage of the star HD80606 as it was transited by its ~4-MJup planet. The observations were performed during the predicted transit windows around 2008 October 25 and 2009 February 14. Our data set is unique in that it simultaneously constrains the duration of the transit and the planet's period. Our Markov Chain Monte Carlo analysis of the light curves, combined with constraints from radial-velocity data, yields system parameters consistent with previously reported values. We find a planet-to-star radius ratio marginally smaller than previously reported, corresponding to a planet radius of Rp = 0.921 +/- 0.036RJup.

A Ring Rearrangement Approach to the Synthesis of Benzo[b]quinolizine and Benzoindolizine Architectures

An efficient ring rearrangement metathesis (RRM) approach to the synthesis of benzo[b]quinolizine and benzoindolizine systems from N-propargyl-phenanthridine derivatives is reported. A Hovel use of flash vacuum pyrolysis (FVP) for the Boc-deprotection of acid-sensitive substrates is also disclosed.

Bacterial dioxygenase- and monooxygenase-catalysed sulfoxidation of benzo[b]thiophenes

Asymmetric heteroatom oxidation of benzo[b]thiophenes to yield the corresponding sulfoxides was catalysed by toluene dioxygenase (TDO), naphthalene dioxygenase (NDO) and styrene monooxygenase (SMO) enzymes present in P. putida mutant and E. coli recombinant whole cells. TDO-catalysed oxidation yielded the relatively unstable benzo[b] thiophene sulfoxide; its dimerization, followed by dehydrogenation, resulted in the isolation of stable tetracyclic sulfoxides as minor products with cis-dihydrodiols being the dominant metabolites. SMO mainly catalysed the formation of enantioenriched benzo[b] thiophene sulfoxide and 2-methyl benzo[b] thiophene sulfoxides which racemized at ambient temperature. The barriers to pyramidal sulfur inversion of 2- and 3-methyl benzo[b] thiophene sulfoxide metabolites, obtained using TDO and NDO as biocatalysts, were found to be ca.: 25-27 kcal mol(-1). The absolute configurations of the benzo[b] thiophene sulfoxides were determined by ECD spectroscopy, X-ray crystallography and stereochemical correlation. A site-directed mutant E. coli strain containing an engineered form of NDO, was found to change the regioselectivity toward preferential oxidation of the thiophene ring rather than the benzene ring.

Blue luminous stars in nearby galaxies: Quantitative spectral analysis of M33 B-type supergiant stars

We present the detailed spectral analysis of a sample of M33 B-type supergiant stars, aimed at the determination of their fundamental parameters and chemical composition. The analysis is based on a grid of non-LTE metal line-blanketed model atmospheres including the effects of stellar winds and spherical extension computed with the code FASTWIND. Surface abundance ratios of C, N, and O are used to discuss the chemical evolutionary status of each individual star. The comparison of observed stellar properties with theoretical predictions of massive star evolutionary models shows good agreement within the uncertainties of the analysis. The spatial distribution of the sample allows us to investigate the existence of radial abundance gradients in the disk of M33. The comparison of stellar and H II region O abundances ( based on direct determinations of the electron temperature of the nebulae) shows good agreement. Using a simple linear radial representation, the stellar oxygen abundances result in a gradient of -0.0145 +/- 0.005 dex arcmin(-1) (or -0.06 +/- 0.02 dex kpc(-1)) up to a distance equal to similar to 1.1 times the isophotal radius of the galaxy. A more complex representation cannot be completely discarded by our stellar sample. The stellar Mg and Si abundances follow the trend displayed by O abundances, although with shallower gradients. These differences in gradient slope cannot be explained at this point. The derived abundances of the three alpha-elements yield solar metallicity in the central regions of the disk of M33. A comparison with recent planetary nebula data from Magrini and coworkers indicates that the disk of M33 has not suffered from a significant O enrichment in the last 3 Gyr.

Space telescope imaging spectrograph ultraviolet spectroscopy of early B supergiants in M31

We analyze Space Telescope Imaging Spectrograph (STIS) spectra in the 1150-1700 Angstrom wavelength range obtained for six early B supergiants in the neighboring galaxy M31. Because of their likely high ( nearly solar) abundance, these stars were originally chosen to be directly comparable to their Galactic counterparts and represent a much needed addition to our current sample of B-type supergiants, in our efforts to study the dependence of the wind momentum-luminosity relationship on spectral type and metallicity. As a first step to determine wind momenta we fit the P Cygni profiles of the resonance lines of N V, Si IV, and C IV with standard methods and derive terminal velocities for all of the STIS targets. From these lines we also derive ionic stellar wind column densities. Our results are compared with those obtained previously in Galactic supergiants and confirm earlier claims of