Development of soluble ester-linked aldehyde polymers for proteomics

High molecular weight hyperbranched polyglycerol (HPG) was selected for development as a soluble polymer support for the targeted selection and release of primary-amine containing peptides from a complex mixture. HPG has been functionalized with ester-linked aldehyde groups that can bind primary-amine containing peptides via a reductive alkylation reaction. Once bound, the high molecular weight of the polymer facilitates separation from a complex peptide mixture by employing either a 30 kDa molecular weight cutoff membrane or precipitation in acetonitrile. Following the removal of unbound peptides and reagents, subsequent hydrolysis of the ester linker releases the bound peptide into solution for analysis by mass spectrometry. Released peptides retain the linker moiety and are therefore characteristically mass-shifted. Four water-soluble cleavable aldehyde polymers (CAP1, CAP2, CAP3, and CAP4) ranging in types of linker groups, length of the linker groups, have been prepared and characterized, each demonstrating the ability to selectively enrich and sequence primary-amine peptides from a complex human proteome containing blocked (dimethylated amine) and unblocked (primary amine) peptides. The polymers have very low nonspecific peptide-binding properties while possessing significantly more reactive groups per milligram of the support than commercially available resins. The polymers exhibit a range of reactivities and binding capacities that depend on the type of linker group between the aldehyde group and the polymer. Using various linker structures, we also probed the mechanism of the observed dehydration of hydrolyzed peptides during matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis.     

Establishment of a real-time PCR method for quantification of geosmin-producing Streptomyces spp. in recirculating aquaculture systems

Geosmin and 2-methylisoborneol (MIB) have been associated with off-flavour problems in fish and seafood products, generating a strong negative impact for aquaculture industries. Although most of the producers of geosmin and MIB have been identified as Streptomyces species or cyanobacteria, Streptomyces spp. are thought to be responsible for the synthesis of these compounds in indoor recirculating aquaculture systems (RAS). The detection of genes involved in the synthesis of geosmin and MIB can be a relevant indicator of the beginning of off-flavour events in RAS. Here, we report a real-time polymerase chain reaction (qPCR) protocol targeting geoA sequences that encode a germacradienol synthase involved in geosmin synthesis. New geoA-related sequences were retrieved from eleven geosmin-producing Actinomycete strains, among them two Streptomyces strains isolated from two RAS. Combined with geoA-related sequences available in gene databases, we designed primers and standards suitable for qPCR assays targeting mainly Streptomyces geoA. Using our qPCR protocol, we succeeded in measuring the level of geoA copies in sand filter and biofilters in two RAS. This study is the first to apply qPCR assays to detect and quantify the geosmin synthesis gene (geoA) in RAS. Quantification of geoA in RAS could permit the monitoring of the level of geosmin producers prior to the occurrence of geosmin production. This information will be most valuable for fish producers to manage further development of off-flavour events.     

Influence of temperature on the compaction of an organic powder and the mechanical strength of tablets

The purpose of this work consists in following the dependence of physical properties on the temperature during the compaction of an organic component. A special thermo-regulated die has been developed to realize uniaxial compression at different constant temperatures. This study has shown that a temperature change modifies the microstructures and the mechanical behaviour of the tablets. The measurement of the tablet porosity during the compression cycle allows us to conclude that temperature influences mainly the phenomena occurring during the isobaric stage of the compression cycle and not the ones during the pressure increase. On the other hand, during the pressure increase, the acoustical activity of the powder is reduced when temperature increases. The tensile strength of tablets realized at different temperatures was also studied and shows a maximum around 60 °C that can be explained by the SEM analysis of the microstructure of the tablets.     

Parents' ethnic-racial socialization practices: A review of research and directions for future study.

Recently, there has been an emergence of literature on the mechanisms through which parents transmit information, values, and perspectives about ethnicity and race to their children, commonly referred to as racial or ethnic socialization. This literature has sought to document the nature of such socialization, its antecedents in parents' and children's characteristics and experiences, and its consequences for children's well-being and development. In this article, the authors integrate and synthesize what is known about racial and ethnic socialization on the basis of current empirical research, examining studies concerning its nature and frequency; its child, parent, and ecological predictors; and its consequences for children's development, including ethnic identity, self-esteem, coping with discrimination, academic achievement, and psychosocial well-being. The authors also discuss conceptual and methodological limitations of the literature and suggest directions for future research. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Increase of aminotransferases

Elevated aminotransferases activities are frequent in medical practice. In acute elevations, the mains causes are generally easily found (viral, drug-induced, toxic, ischemic). In moderate or prolonged elevations, the most frequent causes are steatosis (alcoholic, diabetes, obesity) and chronic hepatitis (viral B, D, C, drug-induced and auto-immune diseases.     

Comparison of the measurements of total IGE using the CAP-RIA technique and the UNICAP-FEIA technique

The contribution of a new method of measurement of total IgE by the Fluoro-Enzymo-Immuno-assay (FEIA) with the assistance of Pharmacia UNICAP automation is very reliable since, when compared with the measurement by the CAP SYSTEM RIA on 144 patients who were clinically allergic (84 with a raised and 60 with a normal IgE titre) the result of a comparative study between the two techniques is statistically highly significant. The two methods were also compared by age groups and these results were also statistically highly significant.     

Photo isomerization of cis-cyclooctene to trans-cyclooctene: Integration of a micro-flow reactor and separation by specific adsorption

Liquid-phase adsorption has hardly been established in micro-flow, although this constitutes an industrially vital method for product separation. A micro-flow UV-photo isomerization process converts cis-cyclooctene partly into trans-cyclooctene, leaving an isomeric mixture. Trans-cyclooctene adsorption and thus separation was achieved in a fixed-bed micro-flow reactor, packed with AgNO₃/SiO₂ powder, while the cis-isomer stays in the flow. The closed-loop recycling-flow has been presented as systemic approach to enrich the trans-cyclooctene from its cis-isomer. In-flow adsorption in recycling-mode has hardly been reported so that a full theoretical study has been conducted. This insight is used to evaluate three process design options to reach an optimum yield of trans-cyclooctene. These differ firstly in the variation of the individual residence times in the reactor and separator, the additional process option of refreshing the adsorption column under use, and the periodicity of the recycle flow.     

Pulsatile Flow in Microfluidic Systems

This review describes the current knowledge and applications of pulsatile flow in microfluidic systems. Elements of fluid dynamics at low Reynolds number are first described in the context of pulsatile flow. Then the practical applications in microfluidic processes are presented: the methods to generate a pulsatile flow, the generation of emulsion droplets through harmonic flow rate perturbation, the applications in mixing and particle separation, and the benefits of pulsatile flow for clog mitigation. The second part of the review is devoted to pulsatile flow in biological applications. Pulsatile flows can be used for mimicking physiological systems, to alter or enhance cell cultures, and for bioassay automation. Pulsatile flows offer unique advantages over a steady flow, especially in microfluidic systems, but also require some new physical insights and more rigorous investigation to fully benefit future applications.