Synthesis, Kinetic Evaluation and Utilization of a Biotinylated Dipeptide Proline Diphenyl Phosphonate for the Disclosure of Dipeptidyl Peptidase IV-like Serine Proteases

In this study, we report on the synthesis, kinetic characterisation, and application of a novel biotinylated and active site-directed inactivator of dipeptidyl peptidase IV (DPP-IV). Thus, the dipeptide-derived proline diphenyl phosphonate NH(2)-Glu(biotinyl-PEG)-Pro(P)(OPh)(2) has been prepared by a combination of classical solution- and solid-phase methodologies and has been shown to be an irreversible inhibitor of porcine DPP-IV, exhibiting an over all second-order rate constant (k(i)/K(i)) for inhibition of 1.57 x 10(3) M(-1) min(-1). This value compares favourably with previously reported rates of inactivation of DPP-IV by dipeptides containing a P(1) proline diphenyl phosphonate grouping [B. Boduszek, J. Oleksyszyn, C.M. Kam, J. Selzler, R.E. Smith, J.C. Powers, Dipeptide phophonates as inhibitors of dipeptidyl peptidase IV, J. Med. Chem. 37 (1994) 3969-3976; B.F. Gilmore, J.F. Lynas, C.J. Scott, C. McGoohan, L. Martin, B. Walker, Dipeptide proline diphenyl phosphonates are potent, irreversible inhibitors of seprase (FAPalpha), Biochem, Biophys. Res. Commun. 346 (2006) 436-446.], thus demonstrating that the incorporation of the side-chain modified (N-biotinyl-3-(2-(2-(3-aminopropyloxy)-ethoxy)-ethoxy)-propyl) glutamic acid residue at the P(2) position is compatible with inhibitor efficacy. The utilisation of this probe for the detection of both purified dipeptidyl peptidase IV and the disclosure of a dipeptidyl peptidase IV-like activity from a clinical isolate of Porphyromonas gingivalis, using established electrophoretic and Western blotting techniques previously developed by our group, is also demonstrated.

The WASP Project and the SuperWASP Cameras

The SuperWASP cameras are wide-field imaging systems at the Observatorio del Roque de los Muchachos on the island of La Palma in the Canary Islands, and at the Sutherland Station of the South African Astronomical Observatory. Each instrument has a field of view of some 482 deg2 with an angular scale of 13.7" pixel-1, and is capable of delivering photometry with accuracy better than 1% for objects having V~7.0-11.5. Lower quality data for objects brighter than V~15.0 are stored in the project archive. The systems, while designed to monitor fields with high cadence, are capable of surveying the entire visible sky every 40 minutes. Depending on the observational strategy, the data rate can be up to 100 Gbytes per night. We have produced a robust, largely automatic reduction pipeline and advanced archive, which are used to serve the data products to the consortium members. The main science aim of these systems is to search for bright transiting exoplanet systems suitable for spectroscopic follow-up observations. The first 6 month season of SuperWASP-North observations produced light curves of ~6.7 million objects with 12.9 billion data points.

The SuperWASP wide-field exoplanetary transit survey: candidates from fields 23 h <RA <03 h

Photometric transit surveys promise to complement the currently known sample of extra-solar planets (ESPs) by providing additional information on the planets and especially their radii. Here, we present ESP candidates from one such survey called, the Wide Angle Search for Planets (WASP) obtained with the SuperWASP wide-field imaging system. Observations were taken with SuperWASP North located in La Palma during the 2004 April to October observing season. The data cover fields between 23 and 03 h in RA at declinations above +12. This amounts to over ~400000 stars with V magnitudes 8-13.5. For the stars brighter than 12.5, we achieve better than 1 per cent photometric precision. Here, we present 41 sources with low-amplitude variability between ~1 and 10 mmag, from which we select 12 with periods between 1.2 and 4.4 d as the most promising ESP candidates. We discuss the properties of these ESP candidates, the expected fraction of transits recovered for our sample and implications for the frequency and detection of hot-Jupiters.

Status of SuperWASP I (La Palma)

SuperWASP is an ultra-wide field (over 300 sq. degrees) photometric survey project designed to monitor stars between 7 - 15 mag to high precision and with high cadence over long (greater than or equal to2 months) timescales. The primary science goal of this project is the detection of exoplanetary transits, as well as NEOs and optical transients. The resulting photometric catalogue will be made public via a web-based interface. The SuperWASP instrument consists of an array of cameras each with a 7.8degrees x 7.8degrees field of view, guided by a robotic fork mount and sited in a fibreglass enclosure at the Observatorio de Roque de los Muchachos (ORM), La Palma, Canary Islands. In this progress report, we describe the specifications of the instrument, its semi-automated operation and pipeline data reduction.

The WASP project in the era of robotic telescope networks

We present the current status of the WASP project, a pair of wide angle photometric telescopes, individually called SuperWASP. SuperWASP-I is located in La Palma, and SuperWASP-II at Sutherland in South Africa. SW-I began operations in April 2004. SW-II is expected to be operational in early 2006. Each SuperWASP instrument consists of up to 8 individual cameras using ultra-wide field lenses backed by high-quality passively cooled CCDs. Each camera covers 7.8 x 7.8 sq degrees of sky, for nearly 500 sq degrees of total sky coverage. One of the current aims of the WASP project is the search for extra-solar planet transits with a focus on brighter stars in the magnitude range similar to 8 to 13. Additionally, WASP will search for, optical transients, track Near-Earth Objects, and study many types of variable stars and extragalactic objects. The collaboration has developed a custom-built reduction pipeline that achieves better than I percent photometric precision. We discuss future goals, which include: nightly on-mountain reductions that could be used to automatically drive alerts via a small robotic telescope network, and possible roles of the WASP telescopes as providers in such a network. Additional technical details of the telescopes, data reduction, and consortium members and institutions can be found on the web site at: http://www.superwasp.org/. (c) 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A fast hybrid algorithm for exoplanetary transit searches

We present a fast and efficient hybrid algorithm for selecting exoplanetary candidates from wide-field transit surveys. Our method is based on the widely used SysRem and Box Least-Squares (BLS) algorithms. Patterns of systematic error that are common to all stars on the frame are mapped and eliminated using the SysRem algorithm. The remaining systematic errors caused by spatially localized flat-fielding and other errors are quantified using a boxcar-smoothing method. We show that the dimensions of the search-parameter space can be reduced greatly by carrying out an initial BLS search on a coarse grid of reduced dimensions, followed by Newton-Raphson refinement of the transit parameters in the vicinity of the most significant solutions. We illustrate the method's operation by applying it to data from one field of the SuperWASP survey, comprising 2300 observations of 7840 stars brighter than V = 13.0. We identify 11 likely transit candidates. We reject stars that exhibit significant ellipsoidal variations caused indicative of a stellar-mass companion. We use colours and proper motions from the Two Micron All Sky Survey and USNO-B1.0 surveys to estimate the stellar parameters and the companion radius. We find that two stars showing unambiguous transit signals pass all these tests, and so qualify for detailed high-resolution spectroscopic follow-up.

The iron abundance of the Magellanic Bridge

High-resolution Hubble Space Telescope ultraviolet spectra for five B-type stars in the Magellanic Bridge and in the Large (LMC) and Small (SMC) Magellanic Clouds have been analysed to estimate their iron abundances. Those for the Clouds are lower than estimates obtained from late-type stars or the optical lines in B-type stars by approximately 0.5 dex. This may be due to systematic errors possibly arising from non-local thermodynamic equilibrium (non-LTE) effects or from errors in the atomic data, as similar low Fe abundances have previously been reported from the analysis of the ultraviolet spectra of Galactic early-type stars. The iron abundance estimates for all three Bridge targets appear to be significantly lower than those found for the SMC and LMC by approximately -0.5 and -0.8 dex, respectively, and these differential results should not be affected by any systematic errors present in the absolute abundance estimates. These differential iron abundance estimates are consistent with the underabundances for C, N, O, Mg and Si of approximately -1.1 dex relative to our Galaxy previously found in our Bridge targets. The implications of these very low metal abundances for the Magellanic Bridge are discussed in terms of metal deficient material being stripped from the SMC.

BACE1 Polymorphisms Do Not Influence Platelet Membrane β-secretase Activity or Genetic Susceptibility for Alzheimer’s Disease in the Northern Irish Population

ß-Site amyloid precursor protein (APP)-cleaving enzyme 1 (BACE1) is a biological and positional candidate gene for Alzheimer’s disease (AD). BACE1 is a protease that catalyses APP cleavage at the ß-secretase site. We evaluated all common and putatively functional polymorphisms in the genomic region encompassing BACE1 for an association with AD, and for functional effects on platelet ß-secretase activity. Tag SNPs (n = 10) derived from phase II of the International HapMap Project, and a nonsynonymous variant, were successfully genotyped in 901 Caucasian individuals from Northern Ireland using Sequenom iPLEX and TaqMan technologies. APOE genotyping was performed by PCR-RFLP. Platelet membrane ß-secretase activity was assayed in a subset of individuals (n = 311). Hardy–Weinberg equilibrium was observed for all variants. Evidence for an association with AD was observed with multi-marker haplotype analyses (P = 0.01), and with rs676134 when stratified for APOE genotype (P = 0.02), however adjusting for multiple testing negated the evidence for association of this variant with AD. ?2 analysis of genotype and allele frequencies in cases versus controls for individual SNPs revealed no evidence for association (5% level). No genetic factors were observed that significantly influenced platelet membrane ß-secretase activity. We have selected an appropriate subset of variants suitable for comprehensive genetic investigation of the BACE1 gene. Our results suggest that common BACE1 polymorphisms and putatively functional variants have no significant influence on genetic susceptibility to AD, or platelet ß-secretase activity, in this Caucasian Northern Irish population.

Testing the possible negative association of type 1 diabetes and atopic disease by analysis of the interleukin 4 receptor gene

Variations in the interleukin 4 receptor A (IL4RA) gene have been reported to be associated with atopy, asthma, and allergy, which may occur less frequently in subjects with type 1 diabetes (T1D). Since atopy shows a humoral immune reactivity pattern, and T1D results from a cellular (T lymphocyte) response, we hypothesised that alleles predisposing to atopy could be protective for T1D and transmitted less often than the expected 50% from heterozygous parents to offspring with T1D. We genotyped seven exonic single nucleotide polymorphisms (SNPs) and the -3223 C>T SNP in the putative promoter region of IL4RA in up to 3475 T1D families, including 1244 Finnish T1D families. Only the -3223 C>T SNP showed evidence of negative association (P=0.014). There was some evidence for an interaction between -3233 C>T and the T1D locus IDDM2 in the insulin gene region (P=0.001 in the combined and P=0.02 in the Finnish data set). We, therefore, cannot rule out a genetic effect of IL4RA in T1D, but it is not a major one.