Synthesis of medium-chain-length polyhydroxyalkanoates in arabidopsis thaliana using intermediates of peroxisomal fatty acid beta-oxidation.

Polyhydroxyalkanoate (PHA) is a family of polymers composed primarily of R-3-hydroxyalkanoic acids. These polymers have properties of biodegradable thermoplastics and elastomers. Medium-chain-length PHAs (MCL-PHAs) are synthesized in bacteria by using intermediates of the beta-oxidation of alkanoic acids. To assess the feasibility of producing MCL-PHAs in plants, Arabidopsis thaliana was transformed with the PhaC1 synthase from Pseudomonas aeruginosa modified for peroxisome targeting by addition of the carboxyl 34 amino acids from the Brassica napus isocitrate lyase. Immunocytochemistry demonstrated that the modified PHA synthase was appropriately targeted to leaf-type peroxisomes in light-grown plants and glyoxysomes in dark-grown plants. Plants expressing the PHA synthase accumulated electron-lucent inclusions in the glyoxysomes and leaf-type peroxisomes, as well as in the vacuole. These inclusions were similar to bacterial PHA inclusions. Analysis of plant extracts by GC and mass spectrometry demonstrated the presence of MCL-PHA in transgenic plants to approximately 4 mg per g of dry weight. The plant PHA contained saturated and unsaturated 3-hydroxyalkanoic acids ranging from six to 16 carbons with 41% of the monomers being 3-hydroxyoctanoic acid and 3-hydroxyoctenoic acid. These results indicate that the beta-oxidation of plant fatty acids can generate a broad range of R-3-hydroxyacyl-CoA intermediates that can be used to synthesize MCL-PHAs.

Postmortem computed tomography angiography, contrast medium administration and toxicological analyses in urine.

Postmortem angiography methods that use water soluble or lipid soluble liquid contrast compounds may potentially modify the composition of fluid-based biological samples and thus influence toxicological findings. In this study, we investigated whether toxicological investigations performed in urine collected prior to and post angiography using Angiofil? mixed with paraffin oil are characterized by different qualitative or quantitative results. In addition, we studied whether diluting samples with 1% and 3% contrast medium solution may modify molecule concentration. A postmortem angiography group consisting of 50 cases and a postmortem group without angiography consisting of 50 cases were formed. In the first group, toxicological investigations were performed in urine samples collected prior to and post angiography as well as in undiluted and diluted samples. In the second group, analyses were performed in undiluted and diluted urine, bile, gastric content, cerebrospinal and pericardial fluids collected during autopsy. The preliminary results indicate that differences may be observed between urine samples collected prior to and post angiography in the number of identified molecules in relation to specific cases. Analyses performed in diluted samples failed to reveal differences that might potentially alter the interpretation of toxicological results in all analyzed specimens for nearly all molecules, except for tetrahydrocannabinol and its metabolites. Though these findings suggest that toxicology might be effectively performed, in very special cases and for a large number of molecules, in biological samples collected after angiography, it remains recommendable to collect biological fluids for toxicology prior to contrast medium injection.

Simultaneous cell morphometry and refractive index measurement with dual-wavelength digital holographic microscopy and dye-enhanced dispersion of perfusion medium.

Digital holographic microscopy (DHM) allows optical-path-difference (OPD) measurements with nanometric accuracy. OPD induced by transparent cells depends on both the refractive index (RI) of cells and their morphology. This Letter presents a dual-wavelength DHM that allows us to separately measure both the RI and the cellular thickness by exploiting an enhanced dispersion of the perfusion medium achieved by the utilization of an extracellular dye. The two wavelengths are chosen in the vicinity of the absorption peak of the dye, where the absorption is accompanied by a significant variation of the RI as a function of the wavelength.

Television before TV : a transnational history of an experimental medium on display, 1928 - 1939

Dès le milieu des années 1920 et le développement des premiers prototypes fonctionnels, la télévision se fait objet d'exposition. La nouvelle technologie est montrée à des foires industrielles et dans des grands magasins, à des expositions universelles et nationales : exposée, la télévision devient un mass media avant même qu'elle ne diffuse des émissions régulières. En étudiant les présentations publiques de la télévision en Allemagne, en Grande-Bretagne et aux Etats-Unis entre 1928 - date de l'ouverture des premières expositions annuelles dans les trois pays - et 1939 - moment de l'interruption de celles-ci suite à l'éclatement de la Deuxième Guerre mondiale - Television before TV montre que les lieux où le médium rencontre son premier public constituent les mêmes espaces qui, rétrospectivement, permettent de comprendre son avènement. Cette recherche propose ainsi de nouvelles pistes pour l'histoire des médias en interrogeant la définition de la télévision en tant que médium du privé. Elle rappelle l'importance historique de l'espace public pour la « vision à distance » et, dépassant la perspective nationale, reconstitue les différences et similarités techniques, médiatiques et institutionnelles de la télévision allemande, américaine et britannique dans l'entre-deux-guerres. Développée dans un contexte qui est simultanément nationaliste et nourri d'échanges et de compétitions internationales, la télévision sert de comparatif dans les rivalités entre pays, mais stimule également la circulation de savoir-faire et de personnes. Sa forte valeur symbolique comme emblème du progrès scientifique et, dans le cas allemand, comme preuve de la modernité du régime national-socialiste se traduit dans les salles d'expositions par des scénographies souvent innovatrices, dont les photographies et descriptions se propagent au-delà de la presse locale. Au centre d'un réseau discursif et représentationnel, les expositions jouent alors un rôle essentiel pour la construction sociale, politique et culturelle de la télévision. Remplaçant l'analyse de textes audiovisuels par l'étude de la machine exposée, ma recherche reformule la question bazinienne de qu'est-ce que la télévision ? pour demander où le médium se situe-t-il ? Cette prémisse méthodologique permet de contourner une évaluation qualitative de la technologie qui déplorerait ses multiples imperfections, et éclaire d'un nouveau jour des dispositifs télévisuels qui, même sans programme, intègrent un univers voué à la culture de consommation et du loisir.

Efficacité a court et moyen termes des traitements d'urgence-crise pour couples et families: résultats et perspectives [Short and medium-term effectiveness of the emergency-crisis treatments for couples and families: results and perspectives].

From 2007 to 2010, the emergency-crisis unit of the Couple and Family Consultation Unit -UCCF (West Psychiatric Service, Prangins Psychiatric Hospital, Psychiatric Department of CHUV) has carried out a research about the relevance and usefulness of emergency-crisis, systemic-oriented treatments, for deeply distressed couples and families. Besides epidemiologic data, we present results demonstrating the efficiency of those treatments, both at short-term and at a one year's range. The global impact of such treatments in terms of public health, but also economical issues, make us believe that they should be fully included in the new trend of psychiatric ambulatory care, into the social net.

Increased flow of fatty acids toward beta-oxidation in developing seeds of Arabidopsis deficient in diacylglycerol acyltransferase activity or synthesizing medium-chain-length fatty acids.

Synthesis of polyhydroxyalkanoates (PHAs) from intermediates of fatty acid beta-oxidation was used as a tool to study fatty acid degradation in developing seeds of Arabidopsis. Transgenic plants expressing a peroxisomal PHA synthase under the control of a napin promoter accumulated PHA in developing seeds to a final level of 0. 06 mg g(-1) dry weight. In plants co-expressing a plastidial acyl-acyl carrier protein thioesterase from Cuphea lanceolata and a peroxisomal PHA synthase, approximately 18-fold more PHA accumulated in developing seeds. The proportion of 3-hydroxydecanoic acid monomer in the PHA was strongly increased, indicating a large flow of capric acid toward beta-oxidation. Furthermore, expression of the peroxisomal PHA synthase in an Arabidopsis mutant deficient in the enzyme diacylglycerol acyltransferase resulted in a 10-fold increase in PHA accumulation in developing seeds. These data indicate that plants can respond to the inadequate incorporation of fatty acids into triacylglycerides by recycling the fatty acids via beta-oxidation and that a considerable flow toward beta-oxidation can occur even in a plant tissue primarily devoted to the accumulation of storage lipids.

The Peroxisomal Enzyme L-PBE Is Required to Prevent the Dietary Toxicity of Medium-Chain Fatty Acids.

Specific metabolic pathways are activated by different nutrients to adapt the organism to available resources. Although essential, these mechanisms are incompletely defined. Here, we report that medium-chain fatty acids contained in coconut oil, a major source of dietary fat, induce the liver ω-oxidation genes Cyp4a10 and Cyp4a14 to increase the production of dicarboxylic fatty acids. Furthermore, these activate all ω- and β-oxidation pathways through peroxisome proliferator activated receptor (PPAR) α and PPARγ, an activation loop normally kept under control by dicarboxylic fatty acid degradation by the peroxisomal enzyme L-PBE. Indeed, L-pbe(-/-) mice fed coconut oil overaccumulate dicarboxylic fatty acids, which activate all fatty acid oxidation pathways and lead to liver inflammation, fibrosis, and death. Thus, the correct homeostasis of dicarboxylic fatty acids is a means to regulate the efficient utilization of ingested medium-chain fatty acids, and its deregulation exemplifies the intricate relationship between impaired metabolism and inflammation.

Debris flow hazard modelling on medium scale: Valtellina di Tirano, Italy

Debris flow hazard modelling at medium (regional) scale has been subject of various studies in recent years. In this study, hazard zonation was carried out, incorporating information about debris flow initiation probability (spatial and temporal), and the delimitation of the potential runout areas. Debris flow hazard zonation was carried out in the area of the Consortium of Mountain Municipalities of Valtellina di Tirano (Central Alps, Italy). The complexity of the phenomenon, the scale of the study, the variability of local conditioning factors, and the lacking data limited the use of process-based models for the runout zone delimitation. Firstly, a map of hazard initiation probabilities was prepared for the study area, based on the available susceptibility zoning information, and the analysis of two sets of aerial photographs for the temporal probability estimation. Afterwards, the hazard initiation map was used as one of the inputs for an empirical GIS-based model (Flow-R), developed at the University of Lausanne (Switzerland). An estimation of the debris flow magnitude was neglected as the main aim of the analysis was to prepare a debris flow hazard map at medium scale. A digital elevation model, with a 10 m resolution, was used together with landuse, geology and debris flow hazard initiation maps as inputs of the Flow-R model to restrict potential areas within each hazard initiation probability class to locations where debris flows are most likely to initiate. Afterwards, runout areas were calculated using multiple flow direction and energy based algorithms. Maximum probable runout zones were calibrated using documented past events and aerial photographs. Finally, two debris flow hazard maps were prepared. The first simply delimits five hazard zones, while the second incorporates the information about debris flow spreading direction probabilities, showing areas more likely to be affected by future debris flows. Limitations of the modelling arise mainly from the models applied and analysis scale, which are neglecting local controlling factors of debris flow hazard. The presented approach of debris flow hazard analysis, associating automatic detection of the source areas and a simple assessment of the debris flow spreading, provided results for consequent hazard and risk studies. However, for the validation and transferability of the parameters and results to other study areas, more testing is needed.

Contrast ultrasound: a simple-to-use phase-shifting medium offers saline infusion sonography-like images.

OBJECTIVE: To test the ability of a novel phase-shifting medium (PSM) to provide sustained distension of the uterine cavity and produce saline infusion sonography (SIS)-like images in a simplified contrast ultrasound procedure. DESIGN: Prospective pilot feasibility trial of a new diagnostic procedure, contrast ultrasound. SETTING: Clinical reproductive endocrine and infertility unit of regional teaching hospital. PATIENT(S): Twenty-six asymptomatic infertile women (group I) and 27 women presenting with dysfunctional uterine bleeding (DUB) who were scheduled for exploratory surgery (group II). INTERVENTION(S): All women who were temporarily on oral contraceptive first had a regular pelvic ultrasound followed by the intrauterine instillation of up to 3 mL PSM, using a regular insemination catheter, after which all instruments were removed and a regular ultrasound was performed again. RESULT(S): In all 53 women, intrauterine instillation of 1-3 mL PSM resulted in a 3-7 mm uterine distension, sufficient to produce SIS-like images of the uterine cavity that lasted 7-10 min. Contrast ultrasound revealed an endometrial polyp in 3 asymptomatic women of group I. In group II. 12 of 14 women (86%) whose vaginal ultrasound were positive or dubious had positive findings with contrast ultrasound; 9 of 12 patients whose vaginal ultrasounds were negative also had positive contrast ultrasound findings. All the positive and negative findings of contrast ultrasound made in group II were confirmed anatomically (sensitivity and specificity of 100%), whereas the correlation for standard vaginal ultrasound was markedly lower at 57.1% and 85.7%, respectively. Most patients (46 of 53) reported no discomfort during or after the procedure, and 7 women described the procedure as mildly uncomfortable. CONCLUSION(S): Contrast ultrasound, a novel simple diagnostic procedure conducted after intrauterine instillation of 1-3 mL PSM using a simple plastic catheter, delivered SIS-quality images in asymptomatic (group I) and symptomatic (group II) patients while retaining the simplicity of standard ultrasound. We therefore foresee broad application of contrast ultrasound for sensitive and specific assessment for uterine pathologies in the physician's office.

A low number of Swiss companies uses nanoparticles

Nanoparticles <100 nanometres are being introduced into industrial processes, but they are suspected to cause similar negative health effects to ambient particles. Poor knowledge about the scale of introduction has not allowed global risk analysis until now. In 2006 a targeted telephone survey among Swiss companies (1) showed the usage of nanoparticles in a few selected companies but did not provide data to extrapolate to the full Swiss workforce. The purpose of the study presented here was to provide a quantitative estimate of the potential occupational exposure to nanoparticles in Swiss industry. Method: A layered representative questionnaire survey among 1626 Swiss companies of the production sector was conducted in 2007. The survey was a written questionnaire, collecting data about the used nanoparticles, the number of potentially exposed persons in the companies and their protection strategy. Results: The response rate of the study was 58.3%. The number of companies estimated to be using nanoparticles in Switzerland was 586 (95% Confidence Interval 145 to 1027). It is estimated that 1309 workers (95% CI 1073 to 1545) do their job in the same room as a nanoparticle application. Personal protection was shown to be the predominant protection means. Such information is valuable for risk evaluation. The low number of companies dealing with nanoparticles in Switzerland suggests that policy makers as well as health, safety and environmental officers within companies can focus their efforts on a relatively small number of companies or workers. The collected data about types of particles and applications may be used for research on prevention strategies and adapted protection means. However, to reflect the most recent trends, the information presented here has to be continuously updated, and a large-scale inventory of the usage should be considered.

Medium-sized neurofilament protein related to maturation of a subset of cortical neurons.

Neurofilaments are typical structures of the neuronal cytoskeleton and participate in the formation and stabilization of the axonal and dendritic architecture. In this study, we have characterized a murine monoclonal antibody, FNP7, that is directed against the medium-sized neurofilament subunit NF-M. This antibody identifies a subset of neurons in the cerebral cortex of various species including human and in organotypic cultures of rat cortex. In the neocortex of all species examined, the antibody labels pyramidal cells in layers III, V, and VI, with a distinctive laminar distribution between architectonic boundaries. In comparison with other antibodies directed against NF-M, the FNP7 antibody identifies on blots two forms of NF-M that appear relatively late during development, at the time when dynamic growth of processes changes to the stabilization of the formed processes. Dephosphorylation with alkaline phosphatase unmasks the site, making it detectable for the FNP7 antibody. The late appearance suggests that the site is present during early development in phosphorylated form and with increasing maturation becomes dephosphorylated, mainly in dendrites. This event may relate to changes in cytoskeleton stability in a late phase of dendritic maturation. Furthermore, mainly corticofugal projections and only few callosal axons are stained, suggesting a differential phosphorylation in a subset of axons. The antibody provides a useful marker to study subsets of pyramidal cells in vivo, in vitro, and under experimental conditions.

Development of a well-defined medium for the in vitro maturation of immature bovine cumulus-oocyte complexes.

PURPOSE: Our purpose was to develop a well-defined medium for the in vitro maturation (IVM) of immature bovine cumulus-oocyte complexes (COC). METHODS: The COC were cultured in the presence of three protein supplementations: fetal bovine serum (FBS), bovine serum albumin, and Synthetic Serum Substitute. The embryos obtained after in vitro fertilization of IVM oocytes were cocultured with Vero cells and their development to the morula and blastocyst stages was studied. RESULTS: When FBS was absent from the IVM medium, a significantly lower fertilization rate was observed, followed by a decrease in the percentage of embryos reaching the blastocyst stage. When FBS was replaced by a defined protein supplementation, the best results were obtained with Synthetic Serum Substitute. CONCLUSIONS: Adequate protein supplementation of the IVM medium optimizes the fertilization rate and the development of bovine IVM oocytes. The implication of these results in the human field is discussed.

Increasing the carbon flux toward synthesis of short-chain-length--medium-chain-length polyhydroxyalkanoate in the peroxisome of Saccharomyces cerevisiae through modification of the beta-oxidation cycle.

Short-chain-length-medium-chain-length polyhydroxyalkanoates were synthesized in Saccharomyces cerevisiae from intermediates of the beta-oxidation cycle by expressing the polyhydroxyalkanoate synthases from Aeromonas caviae and Ralstonia eutropha in the peroxisomes. The quantity of polymer produced was increased by using a mutant of the beta-oxidation-associated multifunctional enzyme with low dehydrogenase activity toward R-3-hydroxybutyryl coenzyme A.