The phylogeny and evolution of Figitidae (Hymenoptera: Cynipoidea)

A phylogeny of the Figitidae (Hymenoptera: Cynipoidea) is presented based on combined analysis of molecular (28S‐D2 and D3, COI and 18S‐E17‐35), morphological and life‐history data. Data are analyzed by parsimony and Bayesian inference methods. Taxon sampling was held at a premium, and the resulting matrix contained 168 terminal taxa representing eight of nine subfamilies (Pycnostigminae not included) and all major subgroups of each subfamily. Alignment of the 28S D2 + D3 gene fragment based on a structural model resulted in the most defendable and least conflicting alignment tested. Melanips, previously classified in Figitinae, was consistently found to be the sister group of the Aspicerinae; Euceroptres, historically classified in Thrasorinae, frequently rendered that subfamily paraphyletic in these analyses. The general evolutionary trend is for early figitids to be parasitoids of gall inducing insects, with later host shifts occurring to exposed hosts associated with aphids. © The Willi Hennig Society 2007.     

Identifying variables responsible for clustering in discriminant analysis of data from infrared microspectroscopy of a biological sample.

In the biomedical field, infrared (IR) spectroscopic studies can involve the processing of data derived from many samples, divided into classes such as category of tissue (e.g., normal or cancerous) or patient identity. We require reliable methods to identify the class-specific information on which of the wavenumbers, representing various molecular groups, are responsible for observed class groupings. Employing a prostate tissue sample divided into three regions (transition zone, peripheral zone, and adjacent adenocarcinoma), and interrogated using synchrotron Fourier-transform IR microspectroscopy, we compared two statistical methods: (a) a new "cluster vector" version of principal component analysis (PCA) in which the dimensions of the dataset are reduced, followed by linear discriminant analysis (LDA) to reveal clusters, through each of which a vector is constructed that identifies the contributory wavenumbers; and (b) stepwise LDA, which exploits the fact that spectral peaks which identify certain chemical bonds extend over several wavenumbers, and which following classification via either one or two wavenumbers, checks whether the resulting predictions are stable across a range of nearby wavenumbers. Stepwise LDA is the simpler of the two methods; the cluster vector approach can indicate which of the different classes of spectra exhibit the significant differences in signal seen at the "prominent" wavenumbers identified. In situations where IR spectra are found to separate into classes, the excellent agreement between the two quite different methods points to what will prove to be a new and reliable approach to establishing which molecular groups are responsible for such separation.     

Assessing spatiotemporal associations in the occurrence of badger–human conflict in England

Examples from a variety of taxa demonstrate that under certain circumstances, the exclusion or translocation of ‘problem’ animals is ineffective in resolving human–wildlife conflicts and may even elicit new problems elsewhere. Damage caused by badger setts (burrows) is an important source of human–wildlife conflict in the UK and is commonly managed by excluding badgers from all or part of problem setts. We used records of licences issued for the management of such problems and a novel statistical approach to assess spatiotemporal associations between problem cases in England from 2002 to 2005. We predicted that management at urban badgers' setts, and particularly exclusion of badgers from urban main setts, would give rise to subsequent problems at focal setts and in neighbouring areas. Frequencies of problems occurring at individual setts were similar in urban and rural areas. In areas neighbouring setts subjected to management action, the background frequency of problems was higher in urban than in rural areas, reflecting the occurrence of problems at a higher proportion of urban setts. The frequency of new cases arising at or in the vicinity of managed setts within a critical time period after management action was not significantly different from the background frequency of problems for any combination of land use, sett type and management approach. This finding suggests that the measures currently employed for managing problem setts do not importantly increase the likelihood of problems reoccurring in the same location or emerging nearby.     

In vitro and in vivo evaluation of 6-aminopyrazolyl-pyridine-3-carbonitriles as JAK2 kinase inhibitors

Synthesis and biological evaluation of a series of 6-aminopyrazolyl-pyridine-3-carbonitriles as JAK2 kinase inhibitors was reported. Biochemical screening, followed by profile optimization, resulted in JAK2 inhibitors exhibiting good kinase selectivity, pharmacokinetic properties, physical properties and pharmacodynamic effects.     

Metabolite cross-feeding enhances virulence in a model polymicrobial infection

Microbes within polymicrobial infections often display synergistic interactions resulting in enhanced pathogenesis; however, the molecular mechanisms governing these interactions are not well understood. Development of model systems that allow detailed mechanistic studies of polymicrobial synergy is a critical step towards a comprehensive understanding of these infections in vivo. In this study, we used a model polymicrobial infection including the opportunistic pathogen Aggregatibacter actinomycetemcomitans and the commensal Streptococcus gordonii to examine the importance of metabolite cross-feeding for establishing co-culture infections. Our results reveal that co-culture with S. gordonii enhances the pathogenesis of A. actinomycetemcomitans in a murine abscess model of infection. Interestingly, the ability of A. actinomycetemcomitans to utilize L-lactate as an energy source is essential for these co-culture benefits. Surprisingly, inactivation of L-lactate catabolism had no impact on mono-culture growth in vitro and in vivo suggesting that A. actinomycetemcomitans L-lactate catabolism is only critical for establishing co-culture infections. These results demonstrate that metabolite cross-feeding is critical for A. actinomycetemcomitans to persist in a polymicrobial infection with S. gordonii supporting the idea that the metabolic properties of commensal bacteria alter the course of pathogenesis in polymicrobial communities.     

Evaluation of large scale quantitative proteomic assay development using peptide affinity-based mass spectrometry

Stable isotope standards and capture by antipeptide antibodies (SISCAPA) couples affinity enrichment of peptides with stable isotope dilution and detection by multiple reaction monitoring mass spectrometry to provide quantitative measurement of peptides as surrogates for their respective proteins. In this report, we describe a feasibility study to determine the success rate for production of suitable antibodies for SISCAPA assays in order to inform strategies for large-scale assay development. A workflow was designed that included a multiplex immunization strategy in which up to five proteotypic peptides from a single protein target were used to immunize individual rabbits. A total of 403 proteotypic tryptic peptides representing 89 protein targets were used as immunogens. Antipeptide antibody titers were measured by ELISA and 220 antipeptide antibodies representing 89 proteins were chosen for affinity purification. These antibodies were characterized with respect to their performance in SISCAPA-multiple reaction monitoring assays using trypsin-digested human plasma matrix. More than half of the assays generated were capable of detecting the target peptide at concentrations of less than 0.5 fmol/μl in human plasma, corresponding to protein concentrations of less than 100 ng/ml. The strategy of multiplexing five peptide immunogens was successful in generating a working assay for 100% of the targeted proteins in this evaluation study. These results indicate it is feasible for a single laboratory to develop hundreds of assays per year and allow planning for cost-effective generation of SISCAPA assays.     

Synthesis and SAR studies of potent imidazopyridine anticoccidial agents

Diaryl imidazo[1,2-a]pyridine derivatives, such as 6a and 7i, have been synthesized and found to be potent inhibitors of parasite PKG activity. The most potent compounds are the 7-isopropylaminomethyl analog 6a and 2-isopropylamino analog 7i. These compounds are also fully active in in vivo assay as anticoccidial agents at 25 ppm in feed.