Production of a broad specificity antibody for the development and validation of an optical SPR screening method for free and intracellular microcystins and nodularin in cyanobacteria cultures

A highly sensitive broad specificity monoclonal antibody was produced and characterised for microcystin detection through the development of a rapid surface plasmon resonance (SPR) optical biosensor based immunoassay. The antibody displayed the following cross-reactivity: MC-LR 100%; MC-RR 108%; MC-YR 68%; MC-LA 69%; MC-LW 71%; MC-LF 68%; and Nodularin 94%. Microcystin-LR was covalently attached to a CM5 chip and with the monoclonal antibody was employed in a competitive 4min injection assay to detect total microcystins in water samples below the WHO recommended limit (1µg/L). A 'total microcystin' level was determined by measuring free and intracellular concentrations in cyanobacterial culture samples as this toxin is an endotoxin. Glass bead beating was used to lyse the cells as a rapid extraction procedure. This method was validated according to European Commission Decision 96/23/EC criteria. The method was proven to measure intracellular microcystin levels, the main source of the toxin, which often goes undetected by other analytical procedures and is advantageous in that it can be used for the monitoring of blooms to provide an early warning of toxicity. It was shown to be repeatable and reproducible, with recoveries from spiked samples ranging from 74 to 123%, and had % CVs below 10% for intra-assay analysis and 15% for inter-assay analysis. The detection capability of the assay was calculated as 0.5ng/mL for extracellular toxins and 0.05ng/mL for intracellular microcystins. A comparison of the SPR method with LC-MS/MS was achieved by testing six Microcystis aeruginosa cultures and this study yielded a correlation R(2) value of 0.9989.

Scale of production and choice of technique in the engineering industries in developing countries

This article examines the relationship between scale of production, optimal choice of technique and costs for three engineering industries: nuts and bolts, iron founding and machine tools. In all three industries costs of production fell as the scale of output increased. This was associated with switches of technique and the spread of fixed costs over a larger number of units. The capital-output ratio fell and labour productivity increased with increases in scale while, in most cases, the capital-labour ratio increased. The implications of these findings are briefly discussed.

Coal combustion residues valorisation:Research and development on compressed brick production

This paper presents the results of an experimental investigation on compressive strength of unfired compressed brick obtained with coal combustion residues (CCRs) produced by the Niger Coal Society. Preliminary physical and optical (XRD and SEM) characterisation of coal slag, including lixiviation tests, have been carried out. Cement powder, lateritic clayey soil and sand have been chosen as stabilizing agents for bricks. 12 dosages have been tested and about 300 bricks have been produced with a hand-operated press. Results show uniaxial compressive strengths (UCSs) ranging from 4 MPa to 27 MPa for the highest cement stabilisation ratio. UCS higher than 7.5 MPa have been observed for stabilisation with 20% of laterite +10% cement after 45 days of curing. Obtained bricks showed good mechanical resistance and low weight. No health threat has been detected for the obtained samples. Study developments are oriented towards the analysis of Pozzolanic properties of CCRs, properties of hydrated lime stabilisation, thermal properties and durability assessment.© 2012 Elsevier Ltd. All rights reserved.

Development in vitro of the neuromusculature of two strigeid trematodes, Apatemon cobitidis proterorhini and Cotylurus erraticus

Confocal microscopy interfaced with cytochemical procedures has been used to monitor development of the major muscle systems and associated serotoninergic (5-HT, 5-hydroxytryptamine) and peptidergic (FaRP, FMRFamide-related peptide) innervation of the strigeid trematodes, Apatemon cobitidis proterorhini and Cotylurus erraticus during cultivation in vitro. Sexually undifferentiated metacercariae were successfully grown to ovigerous adults using tissue culture medium NCTC 135, chicken serum and egg albumen. Eggs were produced after 5 days in culture but had abnormal shells and failed to embryonate. 5-HT and FaRP (the flatworm FaRP, GYIRFamide) were localised immunocytochemically in both central and peripheral nervous systems of developing worms. During cultivation, the central serotoninergic and FaRPergic neuronal pathways of the forebody became more extensive, but retained the same basic orthogonal arrangement as found in the excysted metacercaria. Longitudinal extensor and flexor muscles of the hindbody provide support for the developing reproductive complex. The male reproductive tracts were established in advance (day 3) of those of the female system (day 4); completion of the latter was marked by the appearance of the ootype/egg chamber. The inner longitudinal muscle fibres of the female tract appeared prior to the outer and more densely arranged circular muscles. Circular fibres dominate the muscle complement of both alimentary and reproductive tracts. 5-HT- and GYIRFamide-immunoreactivities were demonstrable in the central nervous system (CNS) and subtegumental parasympathetic nervous system (PNS) throughout the culture period, but innervation of the developing reproductive structures was reactive just for 5-HT. Only at the onset of egg production was FaRP-IR observed in the reproductive system and was expressed only in the innervation of the ootype, a finding consistent with the view that FaRPs may regulate egg assembly in platyhelminths.

First development and characterisation of polyclonal and monoclonal antibodies to the emerging fresh water toxin cylindrospermopsin

As increasing incidences in the occurrence of cylindrospermopsin (CYN) appear, in addition to further research on its toxicological nature, improved rapid methods to detect this toxin are required. Antibody based assays are renowned for their ability to provide rapid, portable, simple to use tests. As yet however there are no publications outlining how an antibody to CYN can be produced. A range of chemical approaches was investigated to synthesise CYN immunogens for antibody production but failed to generate a response. Finally, a modified Mannich reaction for immunogen synthesis was employed to couple the toxin to two carrier proteins. Both protein conjugates were successfully used to raise both polyclonal and monoclonal antibodies of high sensitivity to CYN. These antibodies were characterised employing competitive indirect ELISA and an optical biosensor assay. By ELISA the sensitivity achieved ranged from 27 to 131. pg/mL and by SPR 4.4 to 11.1. ng/mL thus demonstrating that the selection of immunoassay platform is important for the detection level required by the end user for their application. Low cross-reactivity to the much less toxic metabolite deoxyCYN was observed. This is the first reported production of antibodies to this toxin. © 2013 Elsevier B.V.

Suppression of the Insulin Receptors in Adult Schistosoma japonicum Impacts on Parasite Growth and Development: Further Evidence of Vaccine Potential

To further investigate the importance of insulin signaling in the growth, development, sexual maturation and egg production of adult schistosomes, we have focused attention on the insulin receptors (SjIRs) of Schistosoma japonicum, which we have previously cloned and partially characterised. We now show, by Biolayer Interferometry, that human insulin can bind the L1 subdomain (insulin binding domain) of recombinant (r)SjIR1 and rSjIR2 (designated SjLD1 and SjLD2) produced using the Drosophila S2 protein expression system. We have then used RNA interference (RNAi) to knock down the expression of the SjIRs in adult S. japonicum in vitro and show that, in addition to their reduced transcription, the transcript levels of other important downstream genes within the insulin pathway, associated with glucose metabolism and schistosome fecundity, were also impacted substantially. Further, a significant decrease in glucose uptake was observed in the SjIR-knockdown worms compared with luciferase controls. In vaccine/challenge experiments, we found that rSjLD1 and rSjLD2 depressed female growth, intestinal granuloma density and faecal egg production in S. japonicum in mice presented with a low dose challenge infection. These data re-emphasize the potential of the SjIRs as veterinary transmission blocking vaccine candidates against zoonotic schistosomiasis japonica in China and the Philippines.

The Politics of Actually Existing Unsustainability:Human Flourishing in a Climate-Changed, Carbon-Constrained World

Going against both the naive techno-optimist of ‘greening business as usual’ and a resurgent ‘catastrophism’ within green thinking and politics, The Politics of Actually Existing Unsustainability offers an analysis of the causes of unsustainability and diminished human flourishing. The books locates the causes of unsustainability in dominant capitalist modes of production, debt-based consumer culture, the imperative for orthodox economic growth and the dominant ideology of neoclassical economics. It suggests that valuable insights into the causes of and alternatives to unsustainability can be found in a critical embracing of human vulnerability and dependency as both constitutive and ineliminable aspects of what it means to be human. The book defends resilience, the ability to ‘cope with’ rather than somehow ‘solve’ vulnerability. The book offers a trenchant critique of the dominant neoclassical economic ‘groupthink’, viewing it not as some value-neutral form of ‘expert knowledge’, but as a thoroughly ideological ‘common sense’. Outlining a green political economic alternative replacing economic growth with economic security, it argues economic growth has done its work in the minority, affluent world, which should now focus on improving human flourishing, lowering socio-economic equality and fostering solidarity as part of a new re-orientation of public policy. Complementing this, a, ‘green republicanism’ is developed as an innovative and original contribution to contemporary debates on a ‘post-growth’ economy and society. The Politics of Actually Existing Unsustainability draws widely from a range of disciplines and thinkers, from cultural critic Susan Sontag to the critical theory of the Frankfurt School, contemporary debates in green political thinking, and the latest thinking in heterodox and green economics, to produce a highly relevant, timely, and provocatively original statement on the human predicament in the twenty-first century.

Development and Validation of a Procedure for Numerical Vibration Analysis of an Oscillating Wave Surge Converter

During extreme sea states so called impact events can be observed on the wave energy converter Oyster. In small scale experimental tests these impact events cause high frequency signals in the measured load which decrease confidence in the data obtained. These loads depend on the structural dynamics of the model. Amplification of the loads can occur and is transferred through the structure from the point of impact to the load cell located in the foundation. Since the determination of design data and load cases for Wave Energy Converters originate from scale experiments, this lack of confidence has a direct effect on the development.

Numerical vibration analysis is a valuable tool in the research of the structural load response of Oyster to impact events, but must take into account the effect of the surrounding water. This can be done efficiently by adding an added mass distribution, computed with a linearised potential boundary element method. This paper presents the development and validation of a numerical procedure, which couples the OpenSource boundary element code NEMOH with the Finite Element Analysis tool CodeAster. Numerical results of the natural frequencies and mode shapes of the structure under the influence of added mass due to specific structural modes are compared with experimental results.