Physical and immunoaffinity extraction of paralytic shellfish poisoning toxins from cultures of the dinoflagellate Alexandrium tamarense

In the present study the extraction of paralytic shellfish poisoning (PSP) toxins from a toxic strain of the marine dinoflagellate Alexandrium tamarense CCMP-1493 using various mechanical and/or physical procedures was investigated. PBS buffer was investigated as the extraction solvent in order for these procedures to be used directly with immuno-magnetic Ferrospheres-N. The extraction was performed following the determination of when toxin content by the algae was at its highest during batch culture. The methods used for cell lysis and toxin extraction included freeze-thawing, freeze-boiling, steel ball bearing beating, glass bead beating, and sonication. The steel ball bearing beating was determined to release a similar amount of toxin when compared to a modified standard extraction method which was reported to release 100% of toxins from the algal cells and was therefore used in the next phase of the study. This next phase was to determine the feasibility of utilising an antibody coupled to novel magnetic microspheres (Ferrospheres-N) as a simple, rapid immune-capture procedure for PSP toxins extracted from the algae. The effects of increasing mass of Ferrospheres-N on the immuno-capture of the PSP toxins from the toxic algal strain extracts were investigated. Toxin recovery was found to increase when an increasing mass of Ferrospheres-N was used until 96.2% (+/- 1.3 SD) of the toxin extracted from the cells was captured and eluted. Toxin recovery was determined by comparison to an appropriate PSP toxin standard curve following analysis by the AOAC HPLC method. (C) 2011 Elsevier B.V. All rights reserved.

Isolation in cell cultures and initial characterisation of two novel bocavirus species from swine in Northern Ireland

We report the isolation in cell cultures of two novel bocavirus species in pigs from farms in Northern Ireland with clinical postweaning multisystemic wasting syndrome (PMWS). We have designated the isolates as porcine bocavirus-3 (PBoV3) and porcine bocavirus-4 (PBoV4). To date 5082 and 4125 bps of PBoV3 and PBoV4 have been sequenced, respectively. PBoV3 and PBoV4 show nucleotide homology to other known bocaviruses in swine and other organisms. Open reading frame (ORF) analysis has shown that these viruses have a third small ORF, equivalent to the NP1 ORF that distinguishes the bocaviruses from other parvoviruses.

“A Labour Leader in a Frothy Scoundrel”: Farce and Social Justice in the Popular Dramaturgy of George Shiels

In the financially precarious period which followed the partition of Ireland (1922) the Northern Irish playwright George Shiels kept The Abbey Theatre, Dublin, open for business with a series of ‘box-office’ successes. Literary Dublin was not so appreciative of his work as the Abbey audiences dubbing his popular dramaturgy mere ‘kitchen comedy’. However, recent analysts of Irish theatre are beginning to recognise that Shiels used popular theatre methods to illuminate and interrogate instances of social injustice both north and south of the Irish border. In doing so, such commentators have set up a hierarchy between the playwright’s early ‘inferior’ comedies and his later ‘superior’ works of Irish Realism. This article rejects this binary by suggesting that in this early work Shiels’s intent is equally socially critical and that in the plays Paul Twyning, Professor Tim and The Retrievers he is actively engaging with the farcical tradition in order to expose the marginalisation of the landless classes in Ireland in the post-colonial jurisdictions.

Popular Religion in Sixteenth-Century England

An overview of religious life for the majority of the population in England during the century of the Reformation.

Plasmid DNA analysis of Staphylococcus epidermidis isolated from blood and colonization cultures in very low birth weight neonates

We prospectively studied the course of colonization and sepsis with Staphylococcus epidermidis among 29 very low birth weight neonates undergoing prolonged umbilical catheterization. S. epidermidis bacteremia occurred in 7 patients. In 6 bacteremia was preceded by positive colonization cultures. Isolates obtained from nares, base of umbilicus, umbilical catheter entry sites, catheter tips and blood were examined for plasmid DNA profiles. In 4 patients the plasmid profiles of the catheter entry site isolates were identical with those of the blood isolates. In the other 3 bacteremic patients plasmid profiles of the catheter entry site and blood isolates were different. No correlation was observed in the plasmid DNA patterns of isolates obtained from catheter tip cultures as compared to the corresponding blood cultures. The blood isolates from bacteremic patients had different plasmid profiles.

Development of primary human nasal epithelial cell cultures for the study of cystic fibrosis pathophysiology

Cultured primary epithelial cells are used to examine inflammation in cystic fibrosis (CF). We describe a new human model system using cultured nasal brushings. Nasal brushings were obtained from 16 F508del homozygous patients and 11 healthy controls. Cells were resuspended in airway epithelial growth medium and seeded onto collagen-coated flasks and membranes for use in patch-clamp, ion transport, and mediator release assays. Viable cultures were obtained with a 75% success rate from subjects with CF and 100% from control subjects. Amiloride-sensitive epithelial Na channel current of similar size was present in both cell types while forskolin-activated CF transmembrane conductance regulator current was lacking in CF cells. In Ussing chambers, cells from CF patients responded to UTP but not to forskolin. Spontaneous and cytomix-stimulated IL-8 release was similar (stimulated 29,448 ± 9,025 pg/ml; control 16,336 ± 3,308 pg/ml CF; means ± SE). Thus nasal epithelial cells from patients with CF can be grown from nasal brushings and used in electrophysiological and mediator release studies in CF research.

The role of formal structure in liking for popular music

The effect of restructuring the form of three unfamiliar pop/rock songs was investigated in two experiments. In the first experiment, listeners' judgements of the likely location of sections of novel popular songs were explored by requiring participants to place the eight sections (Intro - Verse 1 - Chorus 1 - Verse 2 - Chorus 2 - Bridge (solo) - Chorus 3 - Extro) of the songs into the locations they thought them most likely to occur within the song. Results revealed that participants were able to place the sections in approximately the right location with some accuracy, though they were unable to differentiate between choruses. In Experiment 2, three versions of each of the songs were presented in three different structures: intact (original form), medium restructured (the sections in a moderately changed order), and highly restructured (more severe restructuring). The results show that listeners' judgments of predictability and liking were largely uninfluenced by the restructuring of the songs, in line with findings for classical music. Moment-by-moment liking judgements of the songs demonstrated a change in liking judgements with repeated exposure, though the trend was downwards with repeated exposure rather than upwards. Detailed analysis of moment-by-moment judgements at the ends and beginnings of sections showed that listeners were able to respond quickly to intact songs, but not to restructured songs. The results suggest that concatenism prevails in listening to popular song at the expense of paying attention to larger structural features. © 2012 by the regents of the university of california all rights reserved.