Development of a new portable air sampler based on electrostatic precipitation.

Airborne particles are known to cause illness and to influence meteorological phenomena. It is therefore important to monitor their concentrations and to identify them. A challenge is to collect micro and nanoparticles, microorganisms as well as toxic molecules with a device as simple and small as possible to be used easily and everywhere. Electrostatic precipitation is an efficient method to collect all kinds of airborne particles. Furthermore, this method can be miniaturized. A portable, silent, and autonomous air sampler based on this technology is therefore being developed with the final objective to collect very efficiently airborne pathogens such as supermicron bacteria but also submicron viruses. Particles are collected on a dry surface so they may be concentrated afterwards in a small amount of liquid medium to be analyzed. It is shown that nearly 98 % of airborne particles from 10 nm to 3 µm are collected.

Improvement of yeast biochip sensitivity using multilayer inorganic sol-gel substrates.

Today, microarray fluorescence detection is still limited because a great proportion of hybrids remain undetectable. In this paper we describe sol-gel optical multilayers (stacks of low- and high-index layers) deposited on glass slides which increase the fluorescence of DNA microarrays and favour the detection of fluorescent targets. An alternative to the expensive and time-consuming physical vapour deposition technology is proposed. It is a low-cost sol-gel coating of glass slides, each layer being made by "dipping" (alternatively in SiO2 or TiO2 solutions), "draining and drying". After the selection of the best surface layer of the substrates, the multilayer mirrors modelled for one (Cy3) or two (Cy3 and Cy5) fluorophores are spotted with a series of Yeast probes and compared to similar microarrays on standard glass slides through hybridisation experiments. The fluorescence images of the mirrors show increased signals for all the probes. The enhancement factors determined for Cy3 and for Cy3/Cy5 mirrors (10-12 and 4-5, respectively) are consistent with the initial modelling. This allows the assessment of the basal expression levels of Yeast low-expressed genes. Moreover, these substrates show a noticeable increase in sensitivity for induction/repression ratio measurements in differential gene expression experiments. So, they could be considered as promising tools for the analysis of small biological samples.

DNA microarrays on silicon nanostructures: optimization of the multilayer stack for fluorescence detection.

To improve the sensitivity of fluorescence detection in DNA microarrays, the use of silicon nanostructures based on chemical vapor deposition (CVD) processes adopted for the growth of rough polycrystalline silicon was investigated. These substrates present advantages of two main properties which could lead to an enhancement of the fluorescence detection, i.e. (i) the increase of the available surface area in order to achieve a high loading capacity of biomolecules and (ii) the optimization of the stack of silicon nanostructures support. Indeed, the structures were elaborated on an initial thermal oxide layer and then covered with a silicon oxide layer, obtained by oxidation and allowing the functionalization for the subsequent grafting of DNA probes. Moreover, these oxide layers play a part in the fluorescence detection. The influence of the silicon oxide layer thickness above and below the silicon grains in close relation with the density of nanostructures on the emitted fluorescence was emphasized. This paper presents an experimental characterization of the fluorescence intensity and the optimization of the different layers that composed the substrate used for DNA microarrays. The performances of the microarrays were investigated by means of hybridization experiments using complementary fluorescent labeled-oligonucleotides targets. Our results indicate that an optimized substrate can be designed and that the use of oxidized silicon nanostructures for support of biochip could be a strategy for improving the sensitivity of fluorescence detection.

Measurement of surface concentration of fluorophores by fluorescence fluctuation spectroscopy.

Fluorescence fluctuation spectroscopy is applied to study molecules passing through a small observation volume, usually subjected to diffusive or convective motion in a liquid phase. We suggest that such a technique could be used to measure the areal absolute concentration of fluorophores deposited on a substrate or embedded in a thin film, with a resolution of a few micrometers. The principle is to translate the solid substrate in front of a confocal fluorescence microscope objective and to record the subsequent fluctuations of the fluorescence intensity. The validity of this concept is investigated on model substrates (fluorescent microspheres) and DNA biochips.

Discovery and optimization of 2-aryl oxazolo-pyrimidines as adenosine kinase inhibitors using liquid phase parallel synthesis.

Adenosine kinase inhibition is an attractive therapeutic approach for several conditions for example, neurodegeneration, seizures, ischemia, inflammation and pain. Several nucleosidic and non-nucleosidic inhibitors are available. Using a virtual screening approach, we have discovered that 2-aryl oxazolo-pyrimidines are adenosine kinase inhibitors. Subsequent high throughput derivatization enabled the optimization of this new inhibitor chemotype resulting in highly potent derivatives. A variety of analogues were produced by applying liquid phase parallel synthesis to vary the 7-amino residues as well as the 2-aryl moiety.

Normal and pathological changes in alpha rhythms.

Normal alpha rhythm begins to appear at three months. It was possible to observe successively, the rhythmicity, visual reactivity, occipital topography, and at the end of the first year, the first bursts of 7-8 Hz waves. Evolution of alpha was not linear with the age. Alpha frequency acceleration was important before the age of 11 years. In adults it decreased from 20 to 55, then there was a tendency to increase with aging. Amplitude was highest at 11-12 years, then its tendency was to decrease until 40, and to increase later. In pathology, for instance, alpha could be observed too early in newborn and infants with malformations, in children, in some psychiatric diseases, in adults and old people, alpha was replaced by theta waves in dementia: it seemed to be a very reliable sign, at the beginning of the disease.

[Automatic analysis of sleep-wake states during the first year of life]. / Apport de l'analyse automatique des états veille-sommeil au cours de la première année de la vie.

Automatic analysis on infant sleep tracing was first considered as an aid to the rapid construction of hypnograms. It has long been thought more complicated than in adults, because of the significant changes in electrogenesis during the first year of life, resulting in difficulty to adapt to the criteria of Rechtschaffen and Kales (1968), even if modified, or to those of Anders et al (1971). In fact, these studies have shown that automatic analysis of sleep EEG tracings permit an easier analysis of the changes in electrogenesis according to subject age, sleep stage and time of night. Moreover, automatic analysis of parameters which change with the sleep-wake stages shows the continuity of these changes, and therefore the somewhat arbitrary nature of conventional hypnogram classification. The mathematical treatment of this information facilitates their visualisation, and permits a better analysis of circadian and ultradian variations that could scarcely be formalized by classical hypnogram analysis. In the study of maturation occurring during the first months of life, automatic signal processing will require adaptation that will lead to new forms of reasoning.

Effects of stimulus intensity increase on short-latency somatosensory evoked potentials: application of polynomial curvature coefficients.

We assessed the influence of increasing stimulus intensity from motor threshold to pain threshold on short latency somatosensory evoked potentials recorded over the parietal and frontal scalp in 14 subjects during median nerve stimulation at the wrist. We used the curvature polynomial coefficient to evaluate alterations of the main components. The N20 and P27 curvature coefficients are not modified. The change of the N30 curvature coefficient is the result of shortened P45 latency. The increase of N60 curvature coefficient shows a great interindividual variability, probably due to a central amplification and synchronization or to involvement of nerve fibres with different excitability.

Sleep organization in children at risk for sudden infant death syndrome.

The organization of sleep was studied in four groups of 6-month-old babies: monitored near-miss babies (NM), monitored siblings (HR), nonmonitored siblings (LR), and siblings considered at low risk but monitored because of marked parental anxiety (PA). It was studied using the method of cumulated occurrences and orthogonal polynomial fitting introduced for the analysis of sleep by Gaillard and Martinoli in 1976. No monitoring effect was found. We also found no difference between the groups when the usual sleep scores were used. However, differences were found with the polynomial adjustment method: there was more quiet sleep stage 3 in NM than in any other group. There was less waking state and more paradoxical sleep at the end of the night in PA and NM than in HR or LR babies. This suggests that the differences may not be related so much to real risk as to parental comportment.

[Development of a criterion for the automatic detection of sleep spindles in the infant]. / Mise au point chez le nourrisson d'un critère de détection automatique des fuseaux de sommeil.

The authors present a model for automatic detection of spindles during the different sleep stages. Baseline spindle-free EEG was modelized on a 10 sec recording using an autoregressive model. Spindles were thereafter detected using the quadratic error from this baseline model on successive 1 sec periods. This automatic detection is in a 95.7% agreement with visual analysis.