A Sensitive Assay for Virus Discovery in Respiratory Clinical Samples

In 5–40% of respiratory infections in children, the diagnostics remain negative, suggesting that the patients might be infected with a yet unknown pathogen. Virus discovery cDNA-AFLP (VIDISCA) is a virus discovery method based on recognition of restriction enzyme cleavage sites, ligation of adaptors and subsequent amplification by PCR. However, direct discovery of unknown pathogens in nasopharyngeal swabs is difficult due to the high concentration of ribosomal RNA (rRNA) that acts as competitor. In the current study we optimized VIDISCA by adjusting the reverse transcription enzymes and decreasing rRNA amplification in the reverse transcription, using hexamer oligonucleotides that do not anneal to rRNA. Residual cDNA synthesis on rRNA templates was further reduced with oligonucleotides that anneal to rRNA but can not be extended due to 3′-dideoxy-C6-modification. With these modifications >90% reduction of rRNA amplification was established. Further improvement of the VIDISCA sensitivity was obtained by high throughput sequencing (VIDISCA-454). Eighteen nasopharyngeal swabs were analysed, all containing known respiratory viruses. We could identify the proper virus in the majority of samples tested (11/18). The median load in the VIDISCA-454 positive samples was 7.2 E5 viral genome copies/ml (ranging from 1.4 E3–7.7 E6). Our results show that optimization of VIDISCA and subsequent high-throughput-sequencing enhances sensitivity drastically and provides the opportunity to perform virus discovery directly in patient material

A Behavioral Change Perspective of Maroon Soil Fertility Management in Traditional Shifting Cultivation in Suriname

In Suriname, the Maroons have practiced shifting cultivation for generations, but now the increasing influence of modern society is causing a trend of decreasing fallow periods with potentially adverse effects for the vulnerable tropical soils. Adoption of appropriate soil fertility management (SFM) practices is currently slow. Combining methods from cultural ecology and environmental psychology, this study identifies two groups with divergent behavioral intentions which we term semi-permanent cultivators and shifting cultivators. Semi-permanent cultivators intend to practice more permanent agriculture and experiment individually with plot-level SFM. Shifting cultivators rely on traditional knowledge that is not adequate for their reduced fallow periods, but perceive constraints that prevent them practicing more permanent agriculture. Semi-permanent cultivators act as a strong reference group setting a subjective norm, yet feel no need to exchange knowledge with shifting cultivators who are in danger of feeling marginalized. Drawing on a political ecology perspective, we conclude that cultural ecological knowledge declined due to negative perceptions of external actors setting a strong subjective norm. Semi-permanent cultivators who wish to enter the market economy are most likely to adopt SFM. We conclude that any future SFM intervention must be based on an in-depth understanding of each group’s behavior, in order to avoid exacerbating processes of marginalization

Performance of the CMS Cathode Strip Chambers with Cosmic Rays

The Cathode Strip Chambers (CSCs) constitute the primary muon tracking device in the CMS endcaps. Their performance has been evaluated using data taken during a cosmic ray run in fall 2008. Measured noise levels are low, with the number of noisy channels well below 1%. Coordinate resolution was measured for all types of chambers, and fall in the range 47 microns to 243 microns. The efficiencies for local charged track triggers, for hit and for segments reconstruction were measured, and are above 99%. The timing resolution per layer is approximately 5 ns

Valorisation of sawdust through the combined microwave-assisted hydrothermal pre-treatment and fermentation using an oleaginous yeast

Oleaginous yeast, cultured on second-generation lignocellulosic resources, has the potential to be a key part of the future energy sector. However, the multiple unit operations necessary to produce concentrated hydrolysates, with a minimum of fermentation inhibitors, limit the applicability to date. In this study, a simple microwave-assisted hydrothermal pre-treatment step of oak or beech sawdust was deployed to produce an oligosaccharide-rich hydrolysate. This was then catabolised by the oleaginous yeast, Metschnikowia pulcherrima, avoiding the need for costly enzymatic or further chemical steps in the processing. Up to 85% of the sawdust’s hemicelluloses could be solubilised under these conditions, and 8 g/L DCW yeast with a 42% lipid content produced. While a number of studies have demonstrated that oleaginous yeasts possess high inhibitor tolerance, using this real lignocellulosic hydrolysate, we demonstrate that lipid production is actually very sensitive to inhibitor and carbon availability, and the optimal system is not the one that gives the highest hydrolysate or cell biomass. Indeed, the yeast was shown to detoxify the inhibitors in the process, but at high inhibitor loading, this leads to very poor lipid production, especially at high furfural levels. These findings clearly highlight the importance of considering multiple variables when real, complex lignocellulosic media are involved, tuning process conditions based on the desired fermentation outcomes

Aligning the CMS Muon Chambers with the Muon Alignment System during an Extended Cosmic Ray Run

Peer reviewe

Iron Behaving Badly: Inappropriate Iron Chelation as a Major Contributor to the Aetiology of Vascular and Other Progressive Inflammatory and Degenerative Diseases

The production of peroxide and superoxide is an inevitable consequence of aerobic metabolism, and while these particular "reactive oxygen species" (ROSs) can exhibit a number of biological effects, they are not of themselves excessively reactive and thus they are not especially damaging at physiological concentrations. However, their reactions with poorly liganded iron species can lead to the catalytic production of the very reactive and dangerous hydroxyl radical, which is exceptionally damaging, and a major cause of chronic inflammation. We review the considerable and wide-ranging evidence for the involvement of this combination of (su)peroxide and poorly liganded iron in a large number of physiological and indeed pathological processes and inflammatory disorders, especially those involving the progressive degradation of cellular and organismal performance. These diseases share a great many similarities and thus might be considered to have a common cause (i.e. iron-catalysed free radical and especially hydroxyl radical generation). The studies reviewed include those focused on a series of cardiovascular, metabolic and neurological diseases, where iron can be found at the sites of plaques and lesions, as well as studies showing the significance of iron to aging and longevity. The effective chelation of iron by natural or synthetic ligands is thus of major physiological (and potentially therapeutic) importance. As systems properties, we need to recognise that physiological observables have multiple molecular causes, and studying them in isolation leads to inconsistent patterns of apparent causality when it is the simultaneous combination of multiple factors that is responsible. This explains, for instance, the decidedly mixed effects of antioxidants that have been observed, etc...Comment: 159 pages, including 9 Figs and 2184 reference