The influence of sugar-protein complexes on the thermostability of C-reactive protein (CRP).

The purpose of the present study was to evaluate de influence of protein-sugar complexation on the stability and functionality of C-reactive protein, after exposure to constant high temperatures, in order to develop highly stable positive controls for in-vitro diagnostic tests. C-reactive protein is a plasmatic protein used as a biomarker for the diagnosis of a series of health problems such as ulcerative colitis, cardiovascular diseases, metabolic syndrome, due to its essential role in the evolution of chronic inflammation. The sugar-protein interaction was investigated using steady state and time resolved fluorescence. The results revealed that there are more than two classes of tryptophan, with different degree of accessibility for the quencher molecule. Our study also revealed that sugar-protein complexes have superior thermostability, especially after gamma irradiation at 2 kGy, the protein being stable and functional even after 22 days exposure to 40 °C.

Melittin Induces Local Order Changes in Artificial and Biological Membranes as Revealed by Spectral Analysis of Laurdan Fluorescence.

Antimicrobial peptides (AMPs) are a class of molecules widely used in applications on eukaryotic and prokaryotic cells. Independent of the peptide target, all of them need to first pass or interact with the plasma membrane of the cells. In order to have a better image of the peptide action mechanism with respect to the particular features of the membrane it is necessary to better understand the changes induced by AMPs in the membranes. Laurdan, a lipid membrane probe sensitive to polarity changes in the environment, is used in this study for assessing changes induced by melittin, a well-known peptide, both in model and natural lipid membranes. More importantly, we showed that generalized polarization (GP) values are not always efficient or sufficient to properly characterize the changes in the membrane. We proved that a better method to investigate these changes is to use the previously described log-normal deconvolution allowing us to infer other parameters: the difference between the relative areas of elementary peak (ΔSr), and the ratio of elementary peaks areas (Rs). Melittin induced a slight decrease in local membrane fluidity in homogeneous lipid membranes. The addition of cholesterol stabilizes the membrane more in the presence of melittin. An opposite response was observed in the case of heterogeneous lipid membranes in cells, the local order of lipids being diminished. RS proved to be the most sensitive parameter characterizing the local membrane order, allowing us to distinguish among the responses to melittin of both classes of membrane we investigated (liposomes and cellular membranes). Molecular simulation of the melittin pore in homogeneous lipid bilayer suggests that lipids are more closely packed in the proximity of the melittin pore (a smaller area per lipid), supporting the experimental observation.

Ferrihydrite nanoparticles interaction with model lipid membranes.

In recent years was observed an increased interest towards the use of metal nanoparticles for various biomedical applications, such as therapeutics, delivery systems or imaging. As biological membranes are the first structures with which the nanoparticles interact, it is necessary to understand better the mechanisms governing these interactions. In the present paper we aim to characterize the effect of three different ferrihydrite nanoparticles (simple or doped with cooper or cobalt) on the fluidity of model lipid membranes. First we evaluated the physicochemical properties of the nanoparticles: size and composition. Secondly, their effect on lipid membranes was also evaluated using Laurdan, TMA-DPH and DPH fluorescence. Our results can help better understand the mechanisms involved in nanoparticles and membrane interactions.

Atomic force microscopy study of morphological modifications induced by different decontamination treatments on Escherichia coli.

In this paper we used atomic force microscopy (AFM) to investigate the surface morphology of Escherichia coli, after being subjected to decontamination treatments, at sub-MICs levels (minimal inhibitory concentrations), with different disinfectants used in hospitals, pharmaceutical, food industry and even in our home, as an essential means to prevent the spreading of microorganisms. This article focuses on different morphological modifications adopted by E. coli cells as responses to the different modes of action of these substances. For high-resolution AFM images bacterial cells were immobilized on mica (Muscovite) disks. Each kind of treatment induces its distinct morphological changes, due to different mechanisms of action.

Atmospheric stability effects on potential radiological releases at a nuclear research facility in Romania: Characterising the atmospheric mixing state.

A radon-based nocturnal stability classification scheme is developed for a flat inland site near Bucharest, Romania, characterised by significant local surface roughness heterogeneity, and compared with traditional meteorologically-based techniques. Eight months of hourly meteorological and atmospheric radon observations from a 60 m tower at the IFIN-HH nuclear research facility are analysed. Heterogeneous surface roughness conditions in the 1 km radius exclusion zone around the site hinder accurate characterisation of nocturnal atmospheric mixing conditions using conventional meteorological techniques, so a radon-based scheme is trialled. When the nocturnal boundary layer is very stable, the Pasquill-Gifford "radiation" scheme overestimates the atmosphere's capacity to dilute pollutants with near-surface sources (such as tritiated water vapour) by 20% compared to the radon-based scheme. Under these conditions, near-surface wind speeds drop well below 1 m s(-1) and nocturnal mixing depths vary from ∼ 25 m to less than 10 m above ground level (a.g.l.). Combining nocturnal radon with daytime ceilometer data, we were able to reconstruct the full diurnal cycle of mixing depths. Average daytime mixing depths at this flat inland site range from 1200 to 1800 m a.g.l. in summer, and 500-900 m a.g.l. in winter. Using tower observations to constrain the nocturnal radon-derived effective mixing depth, we were able to estimate the seasonal range in the Bucharest regional radon flux as: 12 mBq m(-2) s(-1) in winter to 14 mBq m(-2) s(-1) in summer.

Fluorescence spectra decomposition by asymmetric functions: Laurdan spectrum revisited.

Due to their asymmetric nature, complex fluorescence spectra of molecules can be analyzed much better by log-normal distributions than by Gaussian ones. So far, the log-normal function has been used for deconvolution of emission spectra of different fluorescent molecules, such as Tryptophan and Prodan, but to our knowledge it is far less used for Laurdan (2-dimethylamino-6-lauroylnaphthalene). In this article, we present the decomposition of Laurdan emission spectra in large unilamellar vesicles using a procedure that relies on the log-normal asymmetric function. The procedure was calibrated using Laurdan spectra in homogeneous solutions of various solvents. Comparing our results with the ones obtained from a Gaussian fit, we show that (i) the position of the elementary peaks (~440 and 490 nm) is preserved in a large range of temperatures that include the main phase transition of lipid bilayer and (ii) the bilayer hydration, as reported by Laurdan, increases approximately 8 times from the gel phase to the liquid crystalline one, a result that fits with other reports, providing a more realistic description. In addition, we propose a new parameter to globally evaluate Laurdan emission spectra with the prospect of acquiring a larger range of values than the classical "generalized polarization".