Analyse par HPLC et CG/SM des constituants du carbone organique dissous (COD), du COD biodégradable (CODB) et des composés organohalogénés (TOX) d'un perméat de nanofiltration

Pour limiter la formation de composés organohalogénés des eaux traitées et la reviviscence bactérienne des réseaux, il est important d'éliminer la majeure partie du carbone organique dissous (COD) et du carbone organique dissous biodégradable (CODB) contenus dans les eaux naturelles. Des travaux récents nous ont permis de montrer que la nanofiltration est une technologie de choix pour répondre à ces contraintes.L'objectif de cet article est de présenter à partir de travaux de laboratoire un inventaire détaillé du carbone organique résiduel d'un perméat prélevé le 21/04/93 sur le prototype industriel de nanofiltration de Méry/Oise en banlieue parisienne. Pour atteindre cet objectif il a été nécessaire de mettre en œuvre des 'techniques analytiques impliquant l'utilisation de la chromatographie liquide haute performance (CLHP) et de la chromatographie en phase gazeuse (CG) soit équipée d'un détecteur à ionisation de flamme (FID) ou d'un détecteur à capture d'électrons (ECD), soit couplée à la spectrométrie de masse (SM).Les résultats obtenus ont montré que le COD du perméat étudié est constitué d'environ 60% d'acides aminés libres et combinés, de 7% d'aldéhydes et de 10 à 20% de composés divers identifiables en CG/SM. Ces derniers composés comprennent majoritairement des acides gras aliphatiques et des acides aromatiques de faibles masses. La concentration de chacun de ces composés a été estimée à 0,3 µg l-¹ C. On peut raisonnablement penser, d'après la bibliographie que les hydrates de carbone (non analysés dans cette étude) représenteraient une part importante de COD du perméat. En outre, cette étude a montré que la part prise par les acides aminés totaux dans le CODB du perméat est importante.Seul le tiers des potentiels de formation d'organohalogénés totaux (PFTOX) a été identifié comme étant des trihaloméhanes (THM) et des acides haloascétiques. Toutefois, étant donné que les acides aminés totaux représentent à eux seuls la quasi totalité de la demande en chlore du perméat, les autres sous-produits de chloration non identifiés seraient probablement des nitriles chlorés, des chloramines et des chloroaldéhydes qui sont parmi les principaux intermédiaires réactionnels de la coloration des acides aminés.Removal of dissolved organic carbon (DOC) and biodegradable dissolved organic carbon (BDOC) is one of the most important means to prevent disinfection by-products (DBPs) formation during water treatment and bacterial regrowth in distribution systems. In previous investigations, the authors have shown that nanofiltration, over nine months of operation at industrial scale in Paris suburbs, was an effective technology to meet the new guidelines concerning chlorine DBPs.This paper reports laboratory investigations aimed to identify and quantify the main organic components included in the low DOC, BDOC and TOXFP (Total - Organohalides Formation Potential) residuals of a nanofiltration permeate sampled on April 21, 1993.Details on DOC, BDOC, organohalides, amino acids and aldehydes analysis procedures were described elsewhere (AGBEKODO et al., 1994). Chlorination was undertaken in potential conditions k: 2.5 mg Cl2/mg DOC, pH=7.5 (phosphate buffer), 72 hours contact time, in dark at 20°C. Haloacetic acids determination consisted (after chlorination in potential conditions) in liquid-liquid extraction, methylation with diazomethane and gas chromatography analysis. Extraction procedure based on XAD8/XAD4 adsorption prior to gas chromatography/Mass Spectrometry (GC/MS) analysis (Fig. 3), allowed a 19000 fold concentration of the permeate. To prevent possible contamination of the permeate, the extraction system consisted of four glass columns and teflon materials. The flow through the columns was performed under high purity nitrogen gas pressure.Analysis involving high performance liquid chromatography (HPLC) and gas chromatography (GC) in combination with Mass Spectrometry (MS), showed that the studied permeate DOC (- 0.15 mg l-¹ c) consisted of amino acids at an average of 60% of DOC, aldehydes (7%) and 10 to 20% of several other compounds (analyzed in GC/MS) including primarily fatty and aromatic acids of low molecular weight (Table 4a and Table 4b). The maximum concentration of each compound (identified hy GC/MS) has been roughly assessed to 0.3 µg l-¹ C. According to literature, sugars represent probably an important portion of the remaining DOC of the permeate. Moreover, the authors have shown that amino acids represented a large portion of permeate BDOC.Only 34% of the total organohalide potentials were identified as trihalomethanes and haloacetic acids. However, since amino acids represent almost the entire chlorine demand of the permeate, the non- identified chlorination DBPs are likely chlorinated nitriles, chloramines and chloraldehydes which are known as the main reactionnal intermediates of aminoacid chlorination

A tyrosine-rich amelogenin peptide promotes neovasculogenesis in vitro and ex vivo

The formation of new blood vessels has been shown to be fundamental in the repair of many damaged tissues, and we have recently shown that the adult human periodontal ligament contains multipotent stem/progenitor cells that are capable of undergoing vasculogenic and angiogenic differentiation in vitro and ex vivo. Enamel matrix protein (EMP) is a heterogeneous mixture of mainly amelogenin-derived proteins produced during tooth development and has been reported to be sometimes effective in stimulating these processes, including in clinical regeneration of the periodontal ligament. However, the identity of the specific bioactive component of EMP remains unclear. In the present study we show that, while the high-molecular-weight Fraction A of enamel matrix derivative (a heat-treated form of EMP) is unable to stimulate the vasculogenic differentiation of human periodontal ligament cells (HPC) in vitro, the low-molecular-weight Fraction C significantly up-regulates the expression of the endothelial markers VEGFR2, Tie-1, Tie-2, VE-cadherin and vWF and markedly increases the internalization of low-density lipoprotein. Furthermore, we also demonstrate, for the first time, that the synthetic homolog of the 45-amino acid tyrosine-rich amelogenin peptide (TRAP) present in Fraction C is likely to be responsible for its vasculogenesis-inducing activity. Moreover, the chemically synthesized TRAP peptide is also shown here to be capable of up-regulating the angiogenic differentiation of the HPC, based on its marked stimulation of in vitro cell migration and tubule formation and of blood vessel formation assay in a chick embryo chorioallantoic membrane model ex vivo. This novel peptide, and modified derivatives, might thereby represent a new class of regenerative drug that has the ability to elicit new blood vessel formation and promote wound healing in vivo. © 2013 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved

Inactivation of aPKCλ Reveals a Context Dependent Allocation of Cell Lineages in Preimplantation Mouse Embryos

BACKGROUND:During mammalian preimplantation development, lineage divergence seems to be controlled by the interplay between asymmetric cell division (once cells are polarized) and positional information. In the mouse embryo, two distinct cell populations are first observed at the 16-cell stage and can be distinguished by both their position (outside or inside) and their phenotype (polarized or non-polarized). Many efforts have been made during the last decade to characterize the molecular mechanisms driving lineage divergence. METHODOLOGY/PRINCIPAL FINDINGS:In order to evaluate the importance of cell polarity in the determination of cell fate we have disturbed the activity of the apical complex aPKC/PAR6 using siRNA to down-regulate aPKClambda expression. Here we show that depletion of aPKClambda results in an absence of tight junctions and in severe polarity defects at the 16-cell stage. Importantly, we found that, in absence of aPKClambda, cell fate depends on the cellular context: depletion of aPKClambda in all cells results in a strong reduction of inner cells at the 16-cell stage, while inhibition of aPKClambda in only half of the embryo biases the progeny of aPKClambda defective blastomeres towards the inner cell mass. Finally, our study points to a role of cell shape in controlling cell position and thus lineage allocation. CONCLUSION:Our data show that aPKClambda is dispensable for the establishment of polarity at the 8-cell stage but is essential for the stabilization of cell polarity at the 16-cell stage and for cell positioning. Moreover, this study reveals that in addition to positional information and asymmetric cell divisions, cell shape plays an important role for the control of lineage divergence during mouse preimplantation development. Cell shape is able to influence both the type of division (symmetric or asymmetric) and the position of the blastomeres within the embryo

Kinetic oxygen measurements by CVC96 in L-929 cell cultures

Generally animal and human cells use oxygen during their whole life. Consequently the oxygen use is a simple indicator to test the vitality of cells. When the vitality decreases by the delivery of toxic substances the decrease can be observed directly by the oxygen-use of the cells. To get fast information of the vitality of cells we have measured the O(2)-tension by testing a new model of a bioreactor, the Cell Vitality Checker 96 (CVC96), in practical application. With this CVC96, soon a simple test will exist for the measurement of the oxygen use. In this respect the question had to be answered whether the use in the laboratory is easy and whether oxygen as a parameter in the vitality test can also be applied in future for problems in the field of material testing

Interaction of enamel matrix proteins with human periodontal ligament cells

Dorothy Hodgkin Postgraduate Award for research studies (jointly funded by the Engineering and Physical Sciences Research Council, UK, and by Institut Straumann) and the Research Discretionary Funds of the Periodontology Unit, UCL Eastman Dental Institute. Financial support was also provided by the NIHR Comprehensive Biomedical Research Centre and by the WCU Program of the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (No. R31-10069)

Regulation of signal duration and the statistical dynamics of kinase activation by scaffold proteins

Scaffolding proteins that direct the assembly of multiple kinases into a spatially localized signaling complex are often essential for the maintenance of an appropriate biological response. Although scaffolds are widely believed to have dramatic effects on the dynamics of signal propagation, the mechanisms that underlie these consequences are not well understood. Here, Monte Carlo simulations of a model kinase cascade are used to investigate how the temporal characteristics of signaling cascades can be influenced by the presence of scaffold proteins. Specifically, we examine the effects of spatially localizing kinase components on a scaffold on signaling dynamics. The simulations indicate that a major effect that scaffolds exert on the dynamics of cell signaling is to control how the activation of protein kinases is distributed over time. Scaffolds can influence the timing of kinase activation by allowing for kinases to become activated over a broad range of times, thus allowing for signaling at both early and late times. Scaffold concentrations that result in optimal signal amplitude also result in the broadest distributions of times over which kinases are activated. These calculations provide insights into one mechanism that describes how the duration of a signal can potentially be regulated in a scaffold mediated protein kinase cascade. Our results illustrate another complexity in the broad array of control properties that emerge from the physical effects of spatially localizing components of kinase cascades on scaffold proteins.Comment: 12 pages, 6 figure

How does neopatrimonialism affect the African state? The case of tax collection in Zambia

Following the neopatrimonialism paradigm, it can be hypothesised that in African states informal politics of the rulers infringe on the collection of taxes and in turn reduce state revenue. This article tests this proposition for the case of Zambia. Neopatrimonial continuity in the country is evidenced by three factors : the concentration of political power, the award of personal favours, and the misuse of state resources. Despite this continuity, the revenue performance increased considerably with the creation of the semi-autonomous Zambia Revenue Authority. Donor pressure has been the most important intervening variable accounting for this improvement. Yet, strengthening the collection of central state revenue has been consistent with a neopatrimonial rationale, and may even have fed neopatrimonialism overall, by providing increased resources for particularistic expenditure