Effects of Hormone Agonists on Sf9 Cells, Proliferation and Cell Cycle Arrest

Methoxyfenozide and methoprene are two insecticides that mimic the action of the main hormones involved in the control of insect growth and development, 20-hydroxyecdysone and juvenile hormone. We investigated their effect on the Spodoptera frugiperda Sf9 cell line. Methoxyfenozide was more toxic than methoprene in cell viability tests and more potent in the inhibition of cellular proliferation. Cell growth arrest occurred in the G2/M phase after a methoprene treatment and more modestly in G1 after methoxyfenozide treatment. Microarray experiments and real-time quantitative PCR to follow the expression of nuclear receptors ultraspiracle and ecdysone receptor were performed to understand the molecular action of these hormone agonists. Twenty-six genes were differentially expressed after methoxyfenozide treatment and 55 genes after methoprene treatment with no gene in common between the two treatments. Our results suggest two different signalling pathways in Sf9 cells

Enhanced Colorimetric Detection on Porous Microarrays Using in Situ Substrate Production

International audienceA new technique is reported for the enhanced colonmetric detection of multiplexed hybridization onto porous membrane-based microarrays. This approach combines the use of horseradish peroxidase (HRP) as a label together with a chromogen substrate and a local production of the hydrogen peroxide required for substrate oxidation. This in situ production of coreagent is obtained using glucose oxidase (GOx) directly immobilized within the microarray porous membrane mesh. The oxidation of glucose by the immobilized GOx produces hydrogen peroxide which itself enables the oxidation of TMB (3,3',5,5'-tetramethylbenzidine) by HRP and yields a blue precipitate on positive spots. Thanks to a coreagent overconcentration within the membrane, this design drastically surpasses the performances of the standard TMB/H(2)O(2) kit used for peroxidase label detection. The obtained target limit of detection is then 50 times lower (20 pM) than the one obtained with the standard kit approach, and the dynamic range expands at least one decade. Furthermore, the developed method was shown to compete well with the widely used alkaline phosphatase-BCIP (5-bromo4-chloro-3-indolyl phosphate)/NBT (nitro blue tetrazolium chloride) readout while minimizing background signal. The method was finally successfully applied to the multiplex detection of single nucleotide polymorphisms (SNPs) in complex PCR samples. The background lowering was impacted here positively on the SNPs' detection by increasing the complementary/noncomplementary signal ratio

Multipurpose high-throughput filtering microarrays (HiFi) for DNA and protein assays

International audienceWe are reporting here a low cost colorimetric device for high-throughput multiplexed blood group genotyping and allergy diagnosis, displayed as an automated 96-well microtiter plate format. A porous polymeric membrane sealed at the bottom of each well accounts for the sensor support. For each sensing unit, a 6 x 6 matrix of specific probes is spotted on the external surface of the membrane resulting in 5 mm(2) microarrays. Thanks to the membrane porosity, reagents dispensed into the well can be eliminated through vacuum soaking. This unusual design drastically reduces the assay background signal. The system was first validated on robust models composed of either two complementary oligonucleotide sequences or one allergen/specific rabbit IgG pair. The quality of both oligonucleotide and protein immobilisation on the membrane substrate was then demonstrated together with the capacity to use the arrayed biomolecules as probes for the quantitative detection of specific targets (respectively complementary oligonucleotide and specific antibody). On the basis of these good results, two multiplex assays were developed for crude biological samples testing, focussing on two human in vitro diagnosis applications: a hybridisation assay for multiplex blood group genotyping and a multiparametric immunoassay for allergy diagnosis. In both cases, the transfer to crude biological samples testing was successful i.e. high signal to noise ratio of the stained membranes, reproducibility and good correlation with results obtained using routine testing procedures. (C) 2010 Elsevier B.V. All rights reserved

Concerns, attitudes, beliefs and information seeking practices with respect to nutrition-related issues: a qualitative study in French pregnant women: A qualitative study in French pregnant women

International audienceBackground: From a life course perspective, pregnancy leads to a rise in nutrition awareness and an increase in information flow in favour of adopting healthier eating behaviours. This qualitative study was designed to better understand the determinants of eating behaviours in French pregnant women by focusing on their concerns, attitudes and beliefs and their nutrition-related information seeking practices. Methods: Seven focus groups were conducted, involving a total of 40 French pregnant women. An inductive thematic approach, adapted from the grounded theory, was adopted to analyse the data. Two major themes were identified: eating behaviour and nutrition-related information behaviour. Results: The eating behaviour theme was divided into four sub-themes using the attribution theory. Three external causes affected the eating behaviour of pregnant women (food restrictions, physiological changes and weight gain), and led to frustration and a perceived loss of control. By contrast the adoption of a healthier diet was perceived as internal by pregnant women, and resulted in self-fulfilment and empowerment regarding the health and the well-being of their baby and themselves, and their weight gain management. Greater attention was paid to nutrition-related information obtained from healthcare providers, the social environment and the mass media. Information was passively absorbed or actively sought by pregnant women, but most was perceived as contradictory, which led to confusion. Conclusion: Pregnancy is accompanied by a rise in nutrition awareness, substantiated by eating behaviour modifications due to external and internal causes. However, conflicts between and within information sources result in confusion that can limit the adoption of healthier eating behaviour