Effects of the vasoactive intestinal peptide (VIP) and related peptides on glioblastoma cell growth in vitro

The growth rate of numerous cancer cell lines is regulated in part by actions of neuropeptides of the vasoactive intestinal peptide (VIP) family, which also includes pituitary adenylate cyclase-activating peptide (PACAP), glucagon, and peptide histidine/isoleucine (PHI). The aim of this work was to investigate the effect of these peptides on the growth of the rat glioblastoma cell line C6 in vitro. We also sought to determine which binding sites were correlated with the effects observed. Proliferation studies performed by means of a CyQuant trade mark assay showed that VIP and PACAP strongly stimulated C6 cell proliferation at most of the concentrations tested, whereas PHI increased cell proliferation only when associated with VIP. Two growth hormone-releasing factor (GRF) derivatives and the VIP antagonist hybrid peptide neurotensin-VIP were able to inhibit VIP-induced cell growth stimulation, even at very low concentrations. Binding experiments carried out on intact cultured C6 cells, using 125I-labeled VIP and PACAP as tracers, revealed that the effects of the peptides on cell growth were correlated with the expression on C6 cells of polyvalent high-affinity VIP-PACAP binding sites and of a second subtype corresponding to very high-affinity VIP-selective binding species. The latter subtype, which interacted poorly with PACAP with a 10,000-fold lower affinity than VIP, might mediate the antagonist effects of neurotensin- VIP and of both GRF derivatives on VIP-induced cell growth stimulation

Brain delivery of vasoactive intestinal peptide (VIP) following nasal administration to rats

The aim of this work was to study in rats the nasal route for the brain delivery of the vasoactive intestinal peptide (VIP) neuropeptide. After evaluating VIP stability in solutions obtained from nasal washes, the effect of formulation parameters (pH 4-9, 0-1% (w/v) lauroylcarnitine (LC), hypo- or isoosmolality) on the brain uptake of intranasally administered VIP (10(-8)M)/125I-VIP (300,000 cpm/ml) was studied, using an in situ perfusion technique. Brain radioactivity distribution was assessed by quantitative autoradiographic analysis. Results were compared to intravenously administered VIP. With a hypotonic formulation at pH 4 containing 0.1% LC and 1% bovine serum albumin, VIP stability was satisfactory and loss by adsorption was minimal. Using this formulation, around 0.11% of initial radioactivity was found in the brain after 30 min perfusion and was located in the olfactory bulbs, the midbrain and the cerebellum. HPLC analysis of brain and blood extracts demonstrated the presence of intact VIP in brain and its complete degradation in the blood compartment. By intravenous administration, no intact VIP was found either in brain or in blood. In conclusion, intact VIP could be delivered successfully to the brain using the intranasal route for administration

Anticancer drug delivery with transferrin targeted polymeric chitosan vesicles

The study reports the initial biological evaluation of targeted polymeric glycol chitosan vesicles as carrier systems for doxorubicin (Dox). Transferrin (Tf) was covalently bound to the Dox-loaded palmitoylated glycol chitosan (GCP) vesicles using dimethylsuberimidate (DMSI). For comparison, glucose targeted niosomes were prepared using N-palmitoyl glucosamine. Biological properties were studied using confocal microscopy, flow cytometry, and cytotoxicity assays as well as a mouse xenograft model. Tf vesicles were taken up rapidly with a plateau after 1-2 h and Dox reached the nucleus after 60-90 min. Uptake was not increased with the use of glucose ligands, but higher uptake and increased cytotoxicity were observed for Tf targeted as compared to GCP Dox alone. In the drug-resistant A2780AD cells and in A431 cells, the relative increase in activity was significantly higher for the Tf-GCP vesicles than would have been expected from the uptake studies. All vesicle formulations had a superior in vivo safety profile compared to the free drug. The in vitro advantage of targeted Tf vesicles did not translate into a therapeutic advantage in vivo. All vesicles reduced tumor size on day 2 but were overall less active than the free drug

Comparison between binary and decimal floating-point numbers

International audienceWe introduce an algorithm to compare a binary floating-point (FP) number and a decimal FP number, assuming the "binary encoding" of the decimal formats is used, and with a special emphasis on the basic interchange formats specified by the IEEE 754-2008 standard for FP arithmetic. It is a two-step algorithm: a first pass, based on the exponents only, quickly eliminates most cases, then, when the first pass does not suffice, a more accurate second pass is performed. We provide an implementation of several variants of our algorithm, and compare them

Dynamic Consolidation of Highly Available Web Applications

Datacenters provide an economical and practical solution for hosting large scale n-tier Web applications. When scalability and high availability are required, each tier can be implemented as multiple replicas, which can absorb extra load and avoid a single point of failure. Realizing these benefits in practice, however, requires that replicas be assigned to datacenter nodes according to certain placement constraints. To provide the required quality of service to all of the hosted applications, the datacenter must consider of all of their specific constraints. When the constraints are not satisfied, the datacenter must quickly adjust the mappings of applications to nodes, taking all of the applications' constraints into account. This paper presents Plasma, an approach for hosting highly available Web applications, based on dynamic consolidation of virtual machines and placement constraint descriptions. The placement constraint descriptions allow the data- center administrator to describe the datacenter infrastructure and each appli- cation administrator to describe his requirements on the VM placement. Based on the descriptions, Plasma continuously optimizes the placement of the VMs in order to provide the required quality of service. Experiments on simulated configurations show that the Plasma reconfiguration algorithm is able to man- age a datacenter with up to 2000 nodes running 4000 VMs with 800 placement constraints. Real experiments on a small cluster of 8 working nodes running 3 instances of the RUBiS benchmarks with a total of 21 VMs show that con- tinuous consolidation is able to reach 85% of the load of a 21 working nodes cluster.Externaliser l'hébergement d'une application Web n-tiers virtualisée dans un centre de données est une solution économiquement viable. Lorsque l'administrateur de l'application considère les problèmes de haute disponibilité tels que le passage à l'échelle et de tolérance aux pannes, chaque machine virtuelle (VM) embarquant un tiers est répliquée plusieurs fois pour absorber la charge et éviter les points de défaillance. Dans la pratique, ces VM doivent être placées selon des contraintes de placement précises. Pour fournir une qualité de service à toutes les applications hébergées, l'administrateur du centre de données doit considérer toutes leurs contraintes. Lorsque des contraintes de placement ne sont plus satisfaites, les VM alors doivent être ré-agencées au plus vite pour retrouver un placement viable. Ce travail est complexe dans un environnement consolidé où chaque nœud peut héberger plusieurs VM. Cet article présente Plasma, un système autonome pour héberger les VM des applications Web haute-disponibilité dans un centre de données utilisant la consolidation dynamique. Par l'intermédiaire de scripts de configuration, les administrateurs des applications décrivent les contraintes de placement de leur VM tandis que l'administrateur système décrit l'infrastructure du centre de données. Grâce à ces descriptions, Plasma optimise en continu le placement des VM pour fournir la qualité de service attendue. Une évaluation avec des données simulées montre que l'algorithme de reconfiguration de Plasma permet de superviser 2000 nœuds hébergeant 4000 VM selon 800 contraintes de placement. Une évaluation sur une grappe de 8 nœuds exécutant 3 instances de l'application RUBiS sur 21 VM montre que la consolidation fournit par Plasma atteint 85% des performances d'une grappe de 21 nœuds

A short upstream promoter region mediates transcriptional regulation of the mouse doublecortin gene in differentiating neurons.

peer reviewedABSTRACT: BACKGROUND: Doublecortin (Dcx), a MAP (Microtubule-Associated Protein), is transiently expressed in migrating and differentiating neurons and thereby characterizes neuronal precursors and neurogenesis in developing and adult neurogenesis. In addition, reduced Dcx expression during development has been related to appearance of brain pathologies. Here, we attempt to unveil the molecular mechanisms controlling Dcx gene expression by studying its transcriptional regulation during neuronal differentiation. RESULTS: To determine and analyze important regulatory sequences of the Dcx promoter, we studied a putative regulatory region upstream from the mouse Dcx coding region (pdcx2kb) and several deletions thereof. These different fragments were used in vitro and in vivo to drive reporter gene expression. We demonstrated, using transient expression experiments, that pdcx2kb is sufficient to control specific reporter gene expression in cerebellar cells and in the developing (E14.5) brain. We determined the temporal profile of Dcx promoter activity during neuronal differentiation of mouse embryonic stem cells (mESC) and found that transcriptional activation of the Dcx gene varies along with neuronal differentiation of mESC. Deletion experiments and sequence comparison of Dcx promoters across rodents, human and chicken revealed the importance of a highly conserved sequence in the proximal region of the promoter required for specific and strong expression in neuronal precursors and young neuronal cells. Further analyses revealed the presence in this short sequence of several conserved, putative transcription factor binding sites: LEF/TCF (Lymphoid Enhancer Factor/T-Cell Factor) which are effectors of the canonical Wnt pathway; HNF6/OC2 (Hepatocyte Nuclear Factor-6/Oncecut-2) members of the ONECUT family; and NF-Y/CAAT (Nuclear Factor-Y). CONCLUSIONS: Studies of Dcx gene regulatory sequences using native, deleted and mutated constructs suggest that fragments located upstream of the Dcx coding sequence are sufficient to induce specific Dcx expression in vitro: in heterogeneous differentiated neurons from mESC, in primary mouse cerebellar neurons (PND3) and in organotypic slices cultures. Furthermore, a region in the 3'-end region of the Dcx promoter is highly conserved across several species and exerts positive control on Dcx transcriptional activation. Together, these results indicate that the proximal 3'-end region of the mouse Dcx regulatory sequence is essential for Dcx gene expression during differentiation of neuronal precursors

Entropy: a Consolidation Manager for Clusters

Clusters provide powerful computing environments, but in practice much of this power goes to waste, due to the static allocation of tasks to nodes, regardless of their changing computational requirements. Consolidation is an approach that migrates tasks within a cluster as their computational requirements change, both to reduce the number of nodes that need to be active and to eliminate temporary overload situations. Previous consolidation strategies have relied on task placement heuristics that use only local optimization and typically do not take migration overhead into account. However, heuristics based on only local optimization may miss the globally optimal solution, resulting in unnecessary resource usage, and the overhead for migration may nullify the benefits of consolidation. In this paper, we propose the Entropy resource manager for homogeneous clusters, which performs consolidation based on constraint programming and takes migration overhead into account. The use of constraint programming allows Entropy to find mappings of tasks to nodes that are better than those found by heuristics based on local optimizations, and that are frequently globally optimal in the number of nodes. Because migration overhead is taken into account, Entropy chooses migrations that can be implemented efficiently, incurring a low performance overhead

Activités Antiproliférative et Antiradicalaire d’extraits Aqueux de Quatre Plantes Médicinales Congolaises

Le but de ce travail est d’évaluer les propriétés antiproliférative et antiradicalaire des extraits aqueux de quatre plantes médicinales congolaises dont Morinda lucida, Klainedoxa gabonensis, Tephrosia vogelii et Nauclea latifolia. L’activité antiproliférative de l’extrait aqueux de chacune des quatre espèces a été évaluée in vitro sur une lignée cellulaire cancéreuse (U87-MG) et une lignée cellulaire normale (Hek-293) en utilisant le test MTT. L’activité antiradicalaire a été évaluée en mesurant la capacité de piégeage du radical DPPH. Des analyses phytochimiques des extraits ont été réalisées par chromatographie sur couche mince et par HPLC-PDA. Les extraits aqueux de Klainedoxa gabonenis et de Tephrosia vogelii ont montré une activité antiproliférative contre les cellules cancéreuses U87-MG avec des valeurs de CI50 inférieures à 90 μg/ml. L’extrait aqueux de Klainedoxa gabonenis a montré également une activité antiradicalaire remarquable (CI50 = 4 ± 0,73 μg/ml) .En plus, le traitement des cellules cancéreuses U87-MG à la fois par l’extrait aqueux de Klainedoxa gabonensis (100 μg/ml) et par un inhibiteur de la protéine MEK (1μM) provoque une suppression totale de la prolifération des cellules U87-MG (glioblastome). L’analyse en HPLC–PDA de l’extrait aqueux Klainedoxa gabonenis a montré la présence des composés de type acide gallique (41, 9 %) et quercétine (2,17 %). Notre étude a permis d’identifier deux plantes médicinales aux propriétés antiprolifératives parmi les quatre plantes médicinales congolaises évaluées dont une possédant à la fois les propriétés antiproliférative et antiradicalaire. The purpose of this work is to evaluate antiprolifertive and DPPH radical scavenging activities of aqueous extracts from Morinda lucida Smith, Klainedoxa gabonensis Pierre ex Engl, Tephrosia vogelii Hook f and Nauclea latifolia Sm. The antiproliferative activity of the aqueous extract of each of the four species was evaluated in vitro on a cancer cell line (U87-MG) and a normal cell line (Hek-293) using the MTT assay. The antiradicalar activity was evaluated by measuring the scavenger capacity of the DPPH radical. Phytochemical analyzes of the extract were performed by thin layer chromatography and HPLC-PDA. The aqueous extracts of Klainedoxa gabonenis and Tephrosia vogelii showed antiproliferative activity against U87-MG cancer cells with IC50 values below 90 μg / ml. The aqueous extract of Klainedoxa gabonenis also showed remarkable antiradical activity (IC50 = 4± 0,73 μg/ml). In addition, the treatment of U87-MG cancer cells by both the aqueous extract of Klainedoxa gabonensis (100 μg / ml) and by an MEK protein inhibitor (1 μM) causes a total suppression of U87-cell proliferation. MG (glioblastoma). HPLC-PDA analysis of the aqueous extract Klainedoxa gabonenis showed the presence of gallic acid compounds (41.9%) and Quercetin (2.17%).: Our study identified two medicinal plants with antiproliferative properties among the four Congolese herbal medicines evaluated, one with both anti-proliferative and antiradical properties

Effect of sonication conditions: solvent, time, temperature and reactor type on the preparation of micron sized vermiculite particles

International audienceThe effects of temperature, time, solvent and sonication conditions under air and Argon are described for the preparation of micron and sub-micron sized vermiculite particles in a double-jacketed Rosett-type or cylindrical reactor. The resulting materials were characterized via X-ray powder diffraction (XRD), Field Emission Scanning Electron Microscopy (FE-SEM), Fourier Transform Infrared (FTIR) Spectroscopy, BET surface area analysis, chemical analysis (elemental analysis), thermogravimetry analysis (TGA) and Laser Granulometry. The sonicated vermiculites displayed modified particle morphologies and reduced sizes (observed by scanning electron microscopy and laser granulometry). Under the conditions used in this work, sub-micron sized particles were obtained after 5 h of sonication, whereas longer times promoted aggregation again. Laser granulometry data revealed also that the smallest particles were obtained at high temperature while it is generally accepted that the mechanical effects of ultrasound are optimum at low temperatures according to physical/chemical properties of the used solvent. X-ray diffraction results indicated a reduction of the crystallite size along the basal direction [001]; but structural changes were not observed. Sonication at different conditions also led to surface modifications of the vermiculite particles brought out by BET surface measurements and Infrared Spectroscopy. The results indicated clearly that the efficiency of ultrasound irradiation was significantly affected by different parameters such as temperature, solvent, type of gas and reactor type