Production and Evaluation of Antibodies and Phage Display-Derived Peptide Ligands for Immunomagnetic Separation of Mycobacterium bovis

This study describes the development and optimization of an immunomagnetic separation (IMS) method to isolate Mycobacterium bovis cells from lymph node tissues. Gamma-irradiated whole M. bovis AF2122/97 cells and ethanol-extracted surface antigens of such cells were used to produce M. bovis-speci?c polyclonal and monoclonal antibodies in rabbits and mice. They were also used to generate M. bovis-speci?c peptide ligands by phage display biopanning. The various antibodies and peptide ligands obtained were used to coat MyOne tosyl-activated Dynabeads (Life Technologies), singly or in combination, and evaluated for IMS. Initially, M. bovis capture from Middlebrook 7H9 broth suspensions (concentration range, 10 to 10⁵ CFU/ml) was evaluated by IMS combined with an M. bovis-speci?c touchdown PCR. IMS-PCR results and, subsequently, IMS-culture results indicated that the beads with greatest immunocapture capability for M. bovis in broth were those coated simultaneously with a monoclonal antibody and a biotinylated 12-mer peptide. These dually coated beads exhibited minimal capture (mean of 0.36% recovery) of 12 other Mycobacterium spp. occasionally encountered in veterinary tuberculosis (TB) diagnostic laboratories. When the optimized IMS method was applied to various M. bovis-spiked lymph node matrices, it demonstrated excellent detection sensitivities (50% limits of detection of 3.16 and 57.7 CFU/ml of lymph node tissue homogenate for IMS-PCR and IMS-culture, respectively). The optimized IMS method therefore has the potential to improve isolation of M. bovis from lymph nodes and hence the diagnosis of bovine tuberculosis.

Laganside: Poetry from Northern Ireland Today

An analysis of Alan Gillis's poem 'Laganside' as exemple of a post-ceasefire Northern Irish poetic.

Magnetic switching of ferroelectric domains at room temperature in multiferroic PZTFT

Single-phase magnetoelectric multiferroics are ferroelectric materials that display some form of magnetism. In addition, magnetic and ferroelectric order parameters are not independent of one another. Thus, the application of either an electric or magnetic field simultaneously alters both the electrical dipole configuration and the magnetic state of the material. The technological possibilities that could arise from magnetoelectric multiferroics are considerable and a range of functional devices has already been envisioned. Realising these devices, however, requires coupling effects to be significant and to occur at room temperature. Although such characteristics can be created in piezoelectric-magnetostrictive composites, to date they have only been weakly evident in single-phase multiferroics. Here in a newly discovered room temperature multiferroic, we demonstrate significant room temperature coupling by monitoring changes in ferroelectric domain patterns induced by magnetic fields. An order of magnitude estimate of the effective coupling coefficient suggests a value of ~1 × 10-7 sm-1.

Topoisomerase IIα promotes activation of RNA polymerase I transcription by facilitating pre-initiation complex formation

Type II DNA topoisomerases catalyse DNA double-strand cleavage, passage and re-ligation to effect topological changes. There is considerable interest in elucidating topoisomerase II roles, particularly as these proteins are targets for anti-cancer drugs. Here we uncover a role for topoisomerase IIa in RNA polymerase I-directed ribosomal RNA gene transcription, which drives cell growth and proliferation and is upregulated in cancer cells. Our data suggest that topoisomerase IIa is a component of the initiation-competent RNA polymerase Iß complex and interacts directly with RNA polymerase I-associated transcription factor RRN3, which targets the polymerase to promoter-bound SL1 in pre-initiation complex formation. In cells, activation of rDNA transcription is reduced by inhibition or depletion of topoisomerase II, and this is accompanied by reduced transient double-strand DNA cleavage in the rDNA-promoter region and reduced pre-initiation complex formation. We propose that topoisomerase IIa functions in RNA polymerase I transcription to produce topological changes at the rDNA promoter that facilitate efficient de novo pre-initiation complex formation.

Evidence for a schizophrenia vulnerability locus on chromosome 8p in the Irish Study of High-Density Schizophrenia Families

This study was an attempt to replicate evidence for a vulnerability locus for schizophrenia and associated disorders in the 8p22-21 region reported by Pulver and colleagues.

CI Aql: a Type Ia supernova progenitor?

If recurrent novae are progenitors of Type Ia supernovae, their white dwarfs must have masses close to the Chandrasekhar limit. The most reliable means of determining white dwarf masses in recurrent novae is dynamically, via radial-velocity and rotational-broadening measurements of the companion star. Such measurements require the system to be both eclipsing and to show absorption features from the secondary star. Prior to the work reported here, the only dynamical mass estimate of a recurrent nova was for U Sco, which has a white dwarf mass of 1.55 +/- 0.24 Msolar (Thoroughgood et al. 2001). We present new time-resolved, intermediate-resolution spectroscopy of the eclipsing recurrent nova CI Aquilae (CI Aql) during quiescence. We find the mass of the white dwarf to be 1.00 +/- 0.14 Msolar and the mass of the secondary star to be 2.32 +/- 0.19 Msolar. We estimate the radius of the secondary to be 2.07 +/- 0.06 Rsolar, implying that it is a slightly-evolved early A-type star. The high mass ratio of q = 2.35 +/- 0.24 and the high secondary-star mass implies that the mass transfer occurs on a thermal timescale. We suggest that CI Aql is rapidly evolving into a supersoft X-ray source, and ultimately may explode as a Type Ia supernova within 10 Myr.

CHARACTERIZATION OF SURFACE-PLASMONS ON METAL-OXIDE-SEMICONDUCTOR STRUCTURES

Using the Otto (prism-air gap-sample) configuration p-polarized light of wavelength 632.8 nm has been coupled with greater than 80% efficiency to surface plasmons on the aluminium electrode of silicon-silicon dioxide-aluminium structures. The results show that if the average power per unit area dissipated on the metal film exceeds approximately 1 mW mm-2, then the coupling gap and thus the characteristics of the surface plasmon resonance are noticeably altered. In modelling the optical response of such systems the inclusion of both a non-uniform air coupling gap and a thin cermet layer at the aluminium surface may be necessary.

Coherence and discontinuity in the scaling of species' distribution patterns

The spatial distribution of a species can be characterized at many different spatial scales, from fine-scale measures of local population density to coarse-scale geographical-range structure. Previous studies have shown a degree of correlation in species' distribution patterns across narrow ranges of scales, making it possible to predict fine-scale properties from coarser-scale distributions. To test the limits of such extrapolation, we have compiled distributional information on 16 species of British plants, at scales ranging across six orders of magnitude in linear resolution (1 in to 100 km). As expected, the correlation between patterns at different spatial scales tends to degrade as the scales become more widely separated. There is, however, an abrupt breakdown in cross-scale correlations across intermediate (ca. 0.5 km) scales, suggesting that local and regional patterns are influenced by essentially non-overlapping sets of processes. The scaling discontinuity may also reflect characteristic scales of human land use in Britain, suggesting a novel method for analysing the 'footprint' of humanity on a landscape.

Sclera as a Surrogate Marker for Determining AGE-Modifications in Bruch's Membrane Using a Raman Spectroscopy-Based Index of Aging

PURPOSE. Raman spectroscopy is an effective probe of advanced glycation end products (AGEs) in Bruch's membrane. However, because it is the outermost layer of the retina, this extracellular matrix is difficult to analyze in vivo with current technology. The sclera shares many compositional characteristics with Bruch's membrane, but it is much easier to access for in vivo Raman analysis. This study investigated whether sclera could act as a surrogate tissue for Raman-based investigation of pathogenic AGEs in Bruch's membrane.

METHODS. Human sclera and Bruch's membrane were dissected from postmortem eyes (n = 67) across a wide age range (33-92 years) and were probed by Raman spectroscopy. The biochemical composition, AGEs, and their age-related trends were determined from data reduction of the Raman spectra and compared for the two tissues.

RESULTS. Raman microscopy demonstrated that Bruch's membrane and sclera are composed of a similar range of biomolecules but with distinct relative quantities, such as in the heme/collagen and the elastin/collagen ratios. Both tissues accumulated AGEs, and these correlated with chronological age (R(2) = 0.824 and R(2) = 0.717 for sclera and Bruch's membrane, respectively). The sclera accumulated AGE adducts at a lower rate than Bruch's membrane, and the models of overall age-related changes exhibited a lower rate (one-fourth that of Bruch's membrane) but a significant increase with age (P <0.05).

CONCLUSIONS. The results suggest that the sclera is a viable surrogate marker for estimating AGE accumulation in Bruch's membrane and for reliably predicting chronological age. These findings also suggest that sclera could be a useful target tissue for future patient-based, Raman spectroscopy studies. (Invest Ophthalmol Vis Sci 2011;52:1593-1598) DOI:10.1167/iovs.10-6554

Calibrated integrated backscatter and myocardial fibrosis in patients undergoing cardiac surgery.

Objective - The reported association between calibrated integrated backscatter (cIB) and myocardial fibrosis is based on study of patients with dilated or hypertrophic cardiomyopathy and extensive (mean 15–34%) fibrosis. Its association with lesser degrees of fibrosis is unknown. We examined the relationship between cIB and myocardial fibrosis in patients with coronary artery disease.

Methods - Myocardial histology was examined in left ventricular epicardial biopsies from 40 patients (29 men and 11 women) undergoing coronary artery bypass graft surgery, who had preoperative echocardiography with cIB measurement.

Results - Total fibrosis (picrosirius red staining) varied from 0.7% to 4%, and in contrast to previous reports, cIB showed weak inverse associations with total fibrosis (r=−0.32, p=0.047) and interstitial fibrosis (r=−0.34, p=0.03). However, cIB was not significantly associated with other histological parameters, including immunostaining for collagens I and III, the advanced glycation end product (AGE) Nε-(carboxymethyl)lysine (CML) and the receptor for AGEs (RAGE). When biomarkers were examined, cIB was weakly associated with log plasma levels of amino-terminal pro-B-type natriuretic peptide (r=0.34, p=0.03), creatinine (r=0.33, p=0.04) and glomerular filtration rate (r=−0.33, p=0.04), and was more strongly associated with log plasma levels of soluble vascular endothelial growth factor receptor-1 (sVEGFR-1) (r=0.44, p=0.01) and soluble RAGE (r=0.53, p=0.002).

Conclusions - Higher cIB was not a marker of increased myocardial fibrosis in patients with coronary artery disease, but was associated with higher plasma levels of sVEGFR-1 and soluble RAGE. The role of cIB as a non-invasive index of fibrosis in clinical studies of patients without extensive fibrosis is, therefore, questionable.

O uso das comunidades virtuais no ensino secundário: o caso do "Palco Virtual" ESEN-Net

A evolução do fluxo de informação e os potenciais da comunicação entre pessoas têm-se revelado deveras importante para o sucesso de diferentes organizações. A escola, enquanto organização social, apresenta uma complexidade natural própria a que se juntam todas as valências de ordem educativa, curricular e pedagógica. Tanto no âmbito educativo como organizacional, as Tecnologias de Informação e Comunicação (TIC) têm vindo a assumir um papel cada vez mais influente e imprescindível. Baseado numa metodologia de trabalho participativo e colaborativo, a construção de “palcos virtuais” resulta de uma análise de requisitos funcionais. Deste modo elabora-se um diagnóstico das necessidades de informação pela replicação das práticas diárias que podem ser quer suportadas, quer melhoradas por esta nova solução, fazendo-se um aproveitamento dos recursos físicos e humanos existentes nas escolas. A partir da iniciativa individual de um professor do grupo de informática da ESEN (Escola Secundária de Emídio Navarro – Viseu), teve início em 1999 o projecto ESEN-Net, como uma proposta de metodologia genérica para a construção de soluções baseadas em intranets para a gestão pedagógica de escolas do ensino secundário. O objectivo central deste trabalho é estudar uma comunidade que desenvolve um projecto de integração das TIC numa escola secundária, no sentido de proporcionar elementos que possam servir como contributos para melhorar a sua organização e, eventualmente, como referência para o desenvolvimento de projectos idênticos noutras escolas. A recolha de dados demonstra que a integração da TIC e o palco virtual ESEN-net veio transformar as práticas diárias na comunidade educativa através da utilização das redes telemáticas como instrumentos de ensino-aprendizagem. As TIC e o palco virtual ESEN-net vieram trazer benefícios à comunidade escolar, facilitando assim a criação e partilha de informação e contribuindo para a criação e desenvolvimento de uma comunidade virtual, onde a construção do saber pode ser feita de uma forma activa e partilhada.