Listeria pathogenesis and molecular virulence determinants

The gram-positive bacterium Listeria monocytogenes is the causative agent of listeriosis, a highly fatal opportunistic foodborne infection. Pregnant women, neonates, the elderly, and debilitated or immunocompromised patients in general are predominantly affected, although the disease can also develop in normal individuals. Clinical manifestations of invasive listeriosis are usually severe and include abortion, sepsis, and meningoencephalitis. Listeriosis can also manifest as a febrile gastroenteritis syndrome. In addition to humans, L. monocytogenes affects many vertebrate species, including birds. Listeria ivanovii, a second pathogenic species of the genus, is specific for ruminants. Our current view of the pathophysiology of listeriosis derives largely from studies with the mouse infection model. Pathogenic listeriae enter the host primarily through the intestine. The liver is thought to be their first target organ after intestinal translocation. In the liver, listeriae actively multiply until the infection is controlled by a cell-mediated immune response. This initial, subclinical step of listeriosis is thought to be common due to the frequent presence of pathogenic L. monocytogenes in food. In normal indivuals, the continual exposure to listerial antigens probably contributes to the maintenance of anti-Listeria memory T cells. However, in debilitated and immunocompromised patients, the unrestricted proliferation of listeriae in the liver may result in prolonged low-level bacteremia, leading to invasion of the preferred secondary target organs (the brain and the gravid uterus) and to overt clinical disease. L. monocytogenes and L. ivanovii are facultative intracellular parasites able to survive in macrophages and to invade a variety of normally nonphagocytic cells, such as epithelial cells, hepatocytes, and endothelial cells. In all these cell types, pathogenic listeriae go through an intracellular life cycle involving early escape from the phagocytic vacuole, rapid intracytoplasmic multiplication, bacterially induced actin-based motility, and direct spread to neighboring cells, in which they reinitiate the cycle. In this way, listeriae disseminate in host tissues sheltered from the humoral arm of the immune system. Over the last 15 years, a number of virulence factors involved in key steps of this intracellular life cycle have been identified. This review describes in detail the molecular determinants of Listeria virulence and their mechanism of action and summarizes the current knowledge on the pathophysiology of listeriosis and the cell biology and host cell responses to Listeria infection. This article provides an updated perspective of the development of our understanding of Listeria pathogenesis from the first molecular genetic analyses of virulence mechanisms reported in 1985 until the start of the genomic era of Listeria research

Functional poly(urethane-imide)s containing Lewis bases for SO2 detection by Love surface acoustic wave gas micro-sensors

Functional polymers containing Lewis bases are known as promising materials for the elaboration of highly sensitive micro-sensors for acid gas detection. In this work, poly(urethane-imide)s (PUIs) with Lewis bases were synthesized in two steps only from various functional diols (N-methyldiethanolamine, N-tert-butyldiethanolamine, N-phenyldiethanolamine and 1,4-diethanolpiperazine) containing one or two tertiary amine groups with different chemical structures. The synthesis led to high yields, high molecular weights and a good control of the PUI amine content. These good film-forming polymers were then used as SO2 sensitive coatings on Love surface acoustic wave (L-SAW) micro-sensors. The physical and mechanical properties required for an optimal micro-sensor design were determined by High Resolution Brillouin Spectroscopy of PUI thin layers. Three-layer micro-sensors were developed by respecting the conditions for the L-SAW generation. The three-layer structure included Quartz ST-90 as the piezoelectric substrate, a ZnO guiding layer and a PUI sensitive layer. These micro-sensors were weakly sensitive to temperature and thus compatible with the targeted application. The experimental results for SO 2 detection showed that all the PUI coatings greatly improved the sensitivity compared to a micro-sensor without polymer coating. The basicity of the amine groups was not determining for the micro-sensor sensitivity to the SO2 acid gas and the key factor was their steric hindrance. Finally, the micro-sensor sensitivity increased with the accessibility of the amine groups in the order Piperazine-diol < tBu-DEA ≅ Ph-DEA ≪ MDEA. © 2013 Elsevier B.V. All rights reserved.Peer Reviewe

New film-forming poly(urethane-amide-imide) block copolymers: Influence of soft block on membrane properties for the purification of a fuel octane enhancer by pervaporation

A new family of eight poly(urethane-amide-imide) (PUAI) block copolymers with the same hard block and different soft blocks were synthesized in two steps from a dianhydride monomer containing amide functions (4,4′-methylene- bis(trimellitic anhydride-N-phenylamide)) and α,ω-dihydroxy telechelic oligomers which varied in both chemical structure (polyethers: PEG, PTMG, PPG; polyester: PCL) and molar weight (MW ≅ 600 or 1000 g/mol). The PUAI were obtained in high yields (ranging from 81 to 98 wt%) and with reduced viscosities which varied from 0.36 to 0.84 dL/g (for C = 1 mg/mL in DMF at 25°C). Their characterization by FTIR and 1H NMR fully confirmed their chemical structure. Their solubility was typically limited to a few wt/vol% even in strong apolar diprotic solvents like DMF and NMP. This particular feature showed the very strong physical cross-linking of their very stiff hard block and enabled to cast membranes capable of withstanding exposure to many common organic solvents. Systematic permeability experiments showed that the PUAI membranes could be used to separate the azeotropic mixture EtOH (20 wt%)/ETBE very easily, with interesting prospects for the purification of ETBE (a fuel octane enhancer used instead of lead derivatives in the European Community). An analysis in terms of structure-property relationships pointed out that the soft block molar weight and polarity were two key parameters for the optimization of selective permeability. The best compromise was obtained with the soft block PEG1000. The corresponding polymer led to performances so far outstanding for polyamideimides with a very high flux of more than 1.1 kg/h m2 for a normalized thickness of 5 μm at 50°C and a selectivity α = 22.7 in the high range for this kind of separation. © 2004 Elsevier Ltd. All rights reserved

Morphology and dynamical behaviour of grafted copolymers

International audienceThe interactions between the main chain and lateral chains in grafted polymers play an important role on their dynamics. This is a complex issue, since several length scales are to be taken into account, from the local intramacromolecular interactions between linked moieties up to intermolecular interactions in phase separated systems. Such phenomena are investigated on the basis of new copolymers obtained by controlled radical polymerization. The effect of two parameters are studied separately, namely (i) the number of grafted chains on the main chain (cellulose acetate) and (ii) the length of grafted moieties (poly(methyl(diethylene glycol)methacrylate)). The dynamics of molecular motions was assessed by Low Frequency Mechanical Spectroscopy and Broad Band Dielectric Spectroscopy. The complex shear and dielectric moduli were measured on thin films (60m thickness) as a function of frequency and temperature. The sensitivity of these relaxation spectroscopy techniques, as compared to calorimetry measurements, allowed to study the effects of (i) the chemical structure of the copolymers and (ii) the microstructure at the nanoscale, as investigated by Small Angle X-ray Scattering. It is shownthat the main parameter is the soft grafted moities content: as expected the molecular mobility increases as the content of soft grafted chains increases. In addition, it is shown that this molecular mobility is drastically depressed by thermal treatment

Genetic analysis of Leishmania parasites in Ecuador : are Leishmania (Viannia) panamensis and Leishmania (V.) guyanensis distinct taxa ?

In the course of an epidemiologic survey in Ecuador, the following collection of #Leishmania stocks was isolated : 28 from patients with clinical signs of leishmaniasis, 2 from sloths, 1 from a dog, and 4 from sand flies. For genetic characterization of these stocks, multilocus enzyme electrophoresis (MLEE) and random amplified polymorphic DNA (RAPD) were used. Twenty six of the 35 stocks were identified as either #Leishmania (V.) panamensis or #L. (V.) guyanensis, 2 stocks were identified as #L. (V.) braziliensis, the 2 stocks from sloths showed specific genotypes, and 5 stocks were characterized as hybrids between #L. (V.) braziliensis and #L. (V.) guyanensis. These data show that genetic diversity of #Leishmania in Ecuador is high and that #L. (V.) panamensis/#guyanensis is the dominant group in this country. The genetic analysis questioned the distinctness between the two species #L. (V.) panamensis and #L. (V.) guyanensis, since MLEE and RAPD data did not indicate that #L. (V.) panamensis and #L. (V.) guyanensis correspond to distinct monophyletic lines. Population genetic analysis performed on the #L. (V.) panamensis/#guyanensis$ group favors the hypothesis of a basically clonal population structure. (Résumé d'auteur

Cellulose acetate graft copolymers with nano-structured architectures: Synthesis and characterization

International audienceCellulose acetate is a very good film-forming polymer with major applications in cigarette filters, photographic films, cosmetics and pharmaceutics formulations and membrane separation processes. Nevertheless, its rigidity and relative hydrophobic character can be limiting drawbacks for some applications. In this work, new cellulose acetate materials with highly flexible and hydrophilic grafts were obtained with different hydrophilic/hydrophobic balances. Cellulose acetate was grafted with methyl diethylene glycol methacrylate (MDEGMA) from brominated macroinitiators by atom transfer radical polymerization (ATRP) in two steps. The first step consisted of introducing ATRP initiator groups on cellulose acetate by reacting hydroxyl side groups with 2-bromoisobutyryl bromide. A preliminary study was then carried out to determine the experimental conditions for the controlled ATRP of MDEGMA homopolymerization in a solvent (cyclopentanone) compatible with cellulose acetate grafting. In these conditions, the MDEGMA homopolymerization followed Hanns Fischer's kinetics model accounting for the radical persistent effect. The ATRP grafting was then investigated for two cellulose acetate macroinitiators differing in the number of their ATRP initiator groups. Two families of graft copolymers with nano-structured architectures were obtained. The first family corresponded to copolymers with a high number of short grafts. The copolymers of the second family had almost the same graft weight fractions but a small number of long grafts. The morphology of the graft copolymers was then investigated by synchrotron X-ray scattering. The most informative results showed that the phase segregation depended upon the number and length of the poly(MDEGMA) grafts. The copolymer with 44 wt.% of long grafts showed a segregated morphology of nano-domains with sharp interfaces and a radius of gyration of 11.5 nm (from Guinier's law). These cellulose acetate copolymers eventually led to strong films with potential applications in membrane separations