SILAC-based proteomic quantification of chemoattractant-induced cytoskeleton dynamics on a second to minute timescale

Cytoskeletal dynamics during cell behaviours ranging from endocytosis and exocytosis to cell division and movement is controlled by a complex network of signalling pathways, the full details of which are as yet unresolved. Here we show that SILAC-based proteomic methods can be used to characterize the rapid chemoattractant-induced dynamic changes in the actin–myosin cytoskeleton and regulatory elements on a proteome-wide scale with a second to minute timescale resolution. This approach provides novel insights in the ensemble kinetics of key cytoskeletal constituents and association of known and novel identified binding proteins. We validate the proteomic data by detailed microscopy-based analysis of in vivo translocation dynamics for key signalling factors. This rapid large-scale proteomic approach may be applied to other situations where highly dynamic changes in complex cellular compartments are expected to play a key role

Knowledge in process? Exploring barriers between epidemiological research and local health policy development

The Redes de Trueque (RT) thrived during the economic crisis of 2001 – 2002 in Argentina and still stand out as one of the largest Complementary Currency System in the world. These local exchange networks reach a large scale during times of severe economic distress, but as large non-state initiatives, they pose a governance problem. Four types of governance systems were structured within the Argentine RT, of varying degrees of sustainability: a) loosely regulated market systems, b) hierarchies, c) associational regional networks, and d) local communities. Based on a four dimensional analytical framework, this paper discusses the rules of governance and sustainability of the governance systems in the RT. It found that some became more sustainable than others in terms of achieving combinations of scale and organisational modes

Stable isotope dilution assay for the accurate determination of mycotoxins in maize by UHPLC-MS/MS

A fast, easy-to-handle and cost-effective analytical method for 11 mycotoxins currently regulated in maize and other cereal-based food products in Europe was developed and validated for maize. The method is based on two extraction steps using different acidified acetonitrile–water mixtures. Separation is achieved using ultrahigh-performance liquid chromatography (UHPLC) by a linear water–methanol gradient. After electrospray ionisation, tandem mass spectrometric detection is performed in dynamic multiple reaction monitoring mode. Since accurate mass spectrometric quantification is hampered by matrix effects, uniformly [13C]-labelled mycotoxins for each of the 11 compounds were added to the sample extracts prior to UHPLC-MS/MS analysis. Method performance parameters were obtained by spiking blank maize samples with mycotoxins before as well as after extraction on six levels in triplicates. The twofold extraction led to total recoveries of the extraction steps between 97% and 111% for all target analytes, including fumonisins. The [13C]-labelled internal standards efficiently compensated all matrix effects in electrospray ionisation, leading to apparent recoveries between 88% and 105% with reasonable additional costs. The relative standard deviations of the whole method were between 4% and 11% for all analytes. The trueness of the method was verified by the measurement of several maize test materials with well-characterized concentrations. In conclusion, the developed method is capable of determining all regulated mycotoxins in maize and presuming similar matrix effects and extraction recovery also in other cereal-based foods

Leukodystrophies: a proposed classification system based on pathological changes and pathogenetic mechanisms

Leukodystrophies are genetically determined disorders characterized by the selective involvement of the central nervous system white matter. Onset may be at any age, from prenatal life to senescence. Many leukodystrophies are degenerative in nature, but some only impair white matter function. The clinical course is mostly progressive, but may also be static or even improving with time. Progressive leukodystrophies are often fatal, and no curative treatment is known. The last decade has witnessed a tremendous increase in the number of defined leukodystrophies also owing to a diagnostic approach combining magnetic resonance imaging pattern recognition and next generation sequencing. Knowledge on white matter physiology and pathology has also dramatically built up. This led to the recognition that only few leukodystrophies are due to mutations in myelin- or oligodendrocyte-specific genes, and many are rather caused by defects in other white matter structural components, including astrocytes, microglia, axons and blood vessels. We here propose a novel classification of leukodystrophies that takes into account the primary involvement of any white matter component. Categories in this classification are the myelin disorders due to a primary defect in oligodendrocytes or myelin (hypomyelinating and demyelinating leukodystrophies, leukodystrophies with myelin vacuolization); astrocytopathies; leuko-axonopathies; microgliopathies; and leuko-vasculopathies. Following this classification, we illustrate the neuropathology and disease mechanisms of some leukodystrophies taken as example for each category. Some leukodystrophies fall into more than one category. Given the complex molecular and cellular interplay underlying white matter pathology, recognition of the cellular pathology behind a disease becomes crucial in addressing possible treatment strategies

Correlation of rheology and light scattering in isotactic polypropylene during early stages of crystallization

ABSTRACT: Structure development during crystallization of isotactic polypropylene (iPP) at low undercooling is studied by small-angle light scattering (SALS) and light transmission. The structure development is related to network formation (gelation) as previously measured on the same sample in dynamic mechanical experiments. Close to the rheologically measured gel point, small-angle light scattering (SALS) patterns in both Hv and Vv modes show circular symmetry; density fluctuations (maximum in Vv invariant) as determined by SALS go through a maximum. Orientation fluctuations develop much more slowly and appear at much later stages of the crystallization process. Growing clusters are characterized by very low anisotropy and low internal crystallinity. The characteristic length scale associated with the cluster at early stages is about 1 ím. At later times, evolution of 2-fold and 4-fold symmetry patterns suggests the development of the anisotropic superstructures. The growth of crystalline clusters leads to a strong increase in turbidity as monitored by light transmission in the Vv mode