Surface plasmon resonance imaging (SPRi) as an alternative technique for rapid and quantitative screening of small molecules, useful in drug discovery

Surface plasmon resonance imaging (SPRi) is a label free technology for biomolecular interaction, which gives access to binding kinetic parameters from real time acquisition. It offers the possibility to test in a single run a large number of interactions, allowing rapid identification of the most suitable compounds toward a given biological entity. Until now, this technique has proven to be relevant for interaction between relatively large molecules (protein, antibodies, DNA) but has not been challenged yet for the screening of small molecules that can be of interested in the field of drug discovery. As a proof a principle, we have used SPRi to screen for interaction of several small molecules, referred to as G4-ligands on G-quadruplex DNA. This technology allowed to easy discrimination of the binding properties of four G4-ligands on quadruplex DNA models.     

Identification and Molecular Cloning of Putative Odorant-Binding Proteins from the American Palm Weevil, Rhynchophorus palmarum L.

We have identified and cloned the cDNAs encoding two odorant-binding proteins (OBPs) from the American palm weevil (APW) Rhynchophorus palmarum (Coleoptera, Curculionidae). Degenerate primers were designed from the N-terminal sequences and were used in polymerase chain reaction (PCR) in order to obtain full-length sequences in both males and females. In both sexes, two different cDNAs were obtained, encoding 123 and 115 amino acid-deduced sequences. Each sequence showed few amino acid differences between the sexes. The proteins were named RpalOBP2 and RpalOBP4 for male, RpalOBP2' and RpalOBP4' for female, with the types 2 and 4 presenting only 34% identities. These proteins shared high identity with previously described coleopteran OBPs. In native gels, RpalOBP2 clearly separated into two bands and RpalOBP4 into three bands, suggesting the presence of several conformational isomers. Thus, OBP diversity in this species may rely on both the presence of OBPs from different classes and the occurrence of isoforms for each OBP.     

Development of a questionnaire to assay recalled liking for salt,sweet and fat

Liking for a sweet, salty or fatty diet may induce overconsumption of simple carbohydrates, sodium or lipids. Measuring overall liking of the corresponding sensory sensations contributes to understanding the determinants of dietary behaviours. However, no standardized validated questionnaire assaying these sensations is currently available. In the present study, we developed a web-based questionnaire, “PrefQuest”, which measures recalled liking for the following four sensations: salt, fat and salt, sweet, fat and sweet. PrefQuest included four types of items: (1) liking for sweet, fatty-sweet and fatty-salty foods, (2) preferences in the level of seasoning by adding salt, sweeteners, or fat, (3) preferences for types of dishes in a restaurant menu and (4) overall questions about sweet-, salt- and fat-related behaviours. A development study (n = 198) demonstrated that PrefQuest was feasible, well-perceived, only lasted about 20 min and that items were repeatable (overall mean intra-class correlation coefficient: 0.77, SD 0.08). Construct validity was then evaluated on a larger population sample (n = 47 803). The underlying structure within each of the four sensations was determined by exploratory factor analysis and then internally validated by confirmatory factor analysis. After a selection of the most relevant items, the salt, fat-and-salt, sweet, and fat-and-sweet scales exhibited a theoretically meaningful factor structure, unidimensional for the salt scale and with interrelated sub-dimensions for the sweet, fat-and-salt, and fat-and-sweet scales. Based on the fat-and-sweet and fat-and-salt scales, a fat model was also unveiled. For each factor, internal consistency as well as convergent and divergent validities were demonstrated. Overall, PrefQuest is an internally valid and original tool that can be used to assay recalled liking for sweet, salt, and fat considered altogether or separately as fat and salt or fat and sweet, and can be applied to large population surveys.     

Road crash fatality rates in France: A comparison of road user types,taking account of travel practices

Background

Travel practices are changing: bicycle and motorized two-wheeler (MTW) use are rising in some of France’s large cities. These are cheaper modes of transport and therefore attractive at a time of economic crisis, but they also allow their users to avoid traffic congestion. At the same time, active transport modes such as walking and cycling are encouraged because they are beneficial to health and reduce pollution. It is therefore important to find out more about the road crash risks of the different modes of transport. To do this, we need to take account of the number of individuals who use each, and, even better, their travel levels.

Method

We estimated the exposure-based fatality rates for road traffic crashes in France, on the basis of the ratio between the number of fatalities and exposure to road accident risk. Fatality data were obtained from the French national police database of road traffic casualties in the period 2007–2008. Exposure data was estimated from the latest national household travel survey (ENTD) which was conducted from April 2007 to April 2008. Three quantities of travel were computed for each mode of transport: (1) the number of trips, (2) the distance traveled and (3) the time spent traveling. Annual fatality rates were assessed by road user type, age and sex.

Results

The overall annual fatality rates were 6.3 per 100 million trips, 5.8 per billion kilometers traveled and 0.20 per million hours spent traveling. The fatality rates differed according to road user type, age and sex. The risk of being killed was 20 to 32 times higher for motorized two-wheeler users than for car occupants. For cyclists, the risk of being killed, both on the basis of time spent traveling and the number of trips was about 1.5 times higher than for car occupants. Risk for pedestrians compared to car occupants was similar according to time spent traveling, lower according to the number of trips and higher according to the distance traveled. People from the 17–20 and 21–29 age groups and those aged 70 and over had the highest rates. Males had higher rates than females, by a factor of between 2 and 3.

Conclusion

When exposure is taken into account, the risks for motorized two-wheeler users are extremely high compared to other types of road user. This disparity can be explained by the combination of speed and a lack of protection (except for helmets). The differential is so great that prevention measures could probably not eliminate it. The question that arises is as follows: with regard to public health, should not the use of MTW, or at least of motorcycles, be deterred? The difference between the fatality risk of cyclists and of car occupants is much smaller (1.5 times higher); besides, there is much room for improvements in cyclist safety, for instance by increasing the use of helmets and conspicuity equipment. Traffic calming could also benefit cyclists, pedestrians and perhaps moped users.     

Bioorthogonal Chemical Reporters for Monitoring Unsaturated Fatty‐Acylated Proteins

Dietary unsaturated fatty acids, such as oleic acid, have been shown to be covalently incorporated into a small subset of proteins, but the generality and diversity of this protein modification has not been studied. We synthesized unsaturated fatty‐acid chemical reporters and determined their protein targets in mammalian cells. The reporters can induce the formation of lipid droplets and be incorporated site‐specifically onto known fatty‐acylated proteins and label many proteins in mammalian cells. Quantitative proteomics analysis revealed that unsaturated fatty acids modify similar protein targets to saturated fatty acids, including several immunity‐associated proteins. This demonstrates that unsaturated fatty acids can directly modify many proteins to exert their unique and often beneficial physiological effects in vivo.