Cephalosporin translocation across enterobacterial OmpF and OmpC channels, a filter across the outer membrane.

Gram-negative porins are the main entry for small hydrophilic molecules. We studied translocation of structurally related cephalosporins, ceftazidime (CAZ), cefotaxime (CTX) and cefepime (FEP). CAZ is highly active on E. coli producing OmpF (Outer membrane protein F) but less efficient on cells expressing OmpC (Outer membrane protein C), whereas FEP and CTX kill bacteria regardless of the porin expressed. This matches with the different capacity of CAZ and FEP to accumulate into bacterial cells as quantified by LC-MS/MS (Liquid Chromatography Tandem Mass Spectrometry). Furthermore, porin reconstitution into planar lipid bilayer and zero current assays suggest permeation of ≈1,000 molecules of CAZ per sec and per channel through OmpF versus ≈500 through OmpC. Here, the instant killing is directly correlated to internal drug concentration. We propose that the net negative charge of CAZ represents a key advantage for permeation through OmpF porins that are less cation-selective than OmpC. These data could explain the decreased susceptibility to some cephalosporins of enterobacteria that exclusively express OmpC porins.

An ultrasensitive LC-MS/MS method with liquid phase extraction to determine paclitaxel in both cell culture medium and lysate promising quantification of drug nanocarriers release in vitro.

The quantification of paclitaxel, a chemotherapy drug used to treat different types of cancers, has been performed from complete cell culture medium and cell lysate samples using a simple liquid-liquid extraction procedure in conjunction with liquid chromatography tandem mass spectrometry (LC-MS/MS). A simple sample preparation using methanol and acetic acid as a weaker acid was applied to avoid paclitaxel destruction and to achieve recovery exceeding 80 % from both matrices spiked with paclitaxel and docetaxel used as internal standard. This rapid, simple, selective and sensitive method enabled the quantification of paclitaxel within the linear range of 1-250nM in culture medium and 5-250nM in cell lysate. The lower limit of quantification was achieved in cell culture medium and cell lysates at 0.2 and 1pmol, respectively. This method was successfully applied to human non-small cell lung carcinoma cells (A549 cells) in order to quantify the amount of paclitaxel in both cell culture medium and lysate after incubation with 5, 50 and 100nM of paclitaxel. This ultra-sensitive method promises the quantification of ultra-low concentrations of paclitaxel released from any nanocarriers, allowing the determination of the kinetic profile of drug release, which is an essential parameter to validate the use of nanocarriers for drug delivery in cancer therapy.

Tubulin isoforms identified in the brain by MALDI in-source decay.

Identification of biomarkers is a major issue for enhancement of chemotherapies. The molecular characterization of tissues necessitates the identification of thousands of biomolecules each participating in physiopathological processes. MALDI in-source decay (ISD) fragmentation has already been proven to be effective for protein characterization. However, the difficulty to identify proteins from complex mixtures such as tissue sections can limit the applications of this technique. In this study, we evidenced that tubulin has an unusual fragmentation pathway in the MALDI source. This striking property allowed the detecting of several mouse brain tubulin isotypes simultaneously by simply using laser fragmentation. Tubulin isoforms are consistent markers of a bad prognosis of solid tumors and could be the target of targeted chemotherapies. Such a direct molecular printout of tubulin in tissues is a milestone that should be useful either at preclinical or clinical stage.

PCBs and DDTs in Stenella coeruleoalba dolphins from the French Mediterranean coastal environment (2007-2009): current state of contamination.

Organochlorinated compounds including PolyChloroBiphenyles, Dichloro-DiphenylTrichloroethan and metabolites are determinated in Stenella coeruleoalba (n = 37) stranded on the french Mediterranean coasts from 2007 till 2009. Studies are carried out on lung, muscle, kidney, liver, and blubber. The sought-after compounds are all detected to variable levels in each tissue and organ. In general, total PCBs are the most abundant, followed by total DDTs. The concentration (in ng g(-1) of lipid weight) in blubber of S. coeruleoalba, varied from 2,052 to 158,992 for PCBs and from 1,120 to 45,779 for DDTs. The ratios DDE/tDDTs are higher than 80% in almost all samples. The overall results of this work, compared to previous studies concerning the Mediterranean Sea, seems to confirm the tendency to a decrease of the contamination by organics compounds for the cetaceans in the Western Mediterranean Sea.

Accumulation and distribution of organochlorines (PCBs and DDTs) in various organs of Stenella coeruleoalba and a Tursiops truncatus from Mediterranean littoral environment (France).

The objective of the present study is to determine the levels of contamination by PCBs, DDT and its metabolites in dolphins failed on the coasts of the Mediterranean sea. Samples are represented by six Stenella coeruleoalba and a Tursiops truncatus collected in 2000 and 2003. The studies are achieved on the blubber, the heart, the liver, the kidney, the muscle and the lung. The concentrations of PCBs and DDT are very high in all tissues and organs analyzed. For the PCBs, the concentrations vary between 43,838 and 110,343 microg/kg lipid basis in the blubber, 601 and 39444 microg/kg dried weight in the liver, 1375 and 34512 microg/kg dried weight in the muscle, 3151 and 17082 microg/kg dried weight in the heart, 674 and 12365 microg/kg dried weight in the kidney and finally between 648 and 4118 microg/kg dried weight in the lung. These values are comparable to those previously obtained in our laboratory and by other authors during the years 1990 on the Mediterranean environment. Significant differences in concentrations are noted in tissues and organs, neither according to the age, nor according to the gender. In all the analyzed samples, the contents in PCBs are higher than those of DDT. The average ratios of pp'-DDE/SigmaDDT are close to 0.6 which shows the metabolization of these compounds along the years. The examination of the profiles of congeners shows that the hexachlorinated molecules are dominating in all tissues and organs which supposes the different animals were especially exposed to Pyralen-type compounds of transformer (Dp6).

Effects of 12 years' operation of a sewage treatment plant on trace metal occurrence within a Mediterranean commercial sponge (Spongia officinalis, Demospongiae).

The present field study uses Spongia officinalis for assessing trace metals occurrence in time and space within Mediterranean rocky communities. Nine sites were selected in the Marseille area for studying spatial trends in 12 metal concentrations. Long term changes in 8 metal concentrations were assessed at sites that had been sampled before and 12 years after the opening of a treatment plant. Spongia officinalis highly concentrated all the trace metal surveyed excepted Hg and Cd. The overall contamination level registered provided a classification of the study sites which is congruent with that given by other studies on pollutant accumulation in neighbouring sandy-bottoms or benthic assemblages. Among the metals studied, Fe, Pb, Cr are those that best highlighted a pollution gradient. In the present study, only Cd concentration did not vary in space. Except for Ni, all pollutant concentrations clearly decreased between 1984 and 1999. This very impressive decrease in heavy metal concentrations within the Marseille area represents an indisputable evidence of the improvement of the seawater quality resulting from 12 years' operation of the Marseille sewage plant. Moreover, the significant decrease also recorded in the reference population at Port-Cros might reflect an overall improvement in the seawater quality of the NW Mediterranean.