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Sample records for active zone molecule

  1. Dynamics of Activated Molecules

    Mullin, Amy S. [Univ. of Maryland, College Park, MD (United States)

    2016-11-16

    Experimental studies have been performed to investigate the collisional energy transfer processes of gas-phase molecules that contain large amounts of internal energy. Such molecules are prototypes for molecules under high temperature conditions relevant in combustion and information about their energy transfer mechanisms is needed for a detailed understanding and modeling of the chemistry. We use high resolution transient IR absorption spectroscopy to measure the full, nascent product distributions for collisions of small bath molecules that relax highly vibrationally excited pyrazine molecules with E=38000 cm-1 of vibrational energy. To perform these studies, we developed new instrumentation based on modern IR light sources to expand our experimental capabilities to investigate new molecules as collision partners. This final report describes our research in four areas: the characterization of a new transient absorption spectrometer and the results of state-resolved collision studies of pyrazine(E) with HCl, methane and ammonia. Through this research we have gained fundamental new insights into the microscopic details of relatively large complex molecules at high energy as they undergo quenching collisions and redistribute their energy.

  2. Presynaptic active zone density during development and synaptic plasticity.

    Gwenaëlle L Clarke

    2012-02-01

    Full Text Available Neural circuits transmit information through synapses, and the efficiency of synaptic transmission is closely related to the density of presynaptic active zones, where synaptic vesicles are released. The goal of this review is to highlight recent insights into the molecular mechanisms that control the number of active zones per presynaptic terminal (active zone density during developmental and stimulus-dependent changes in synaptic efficacy. At the neuromuscular junctions (NMJs, the active zone density is preserved across species, remains constant during development, and is the same between synapses with different activities. However, the NMJ active zones are not always stable, as exemplified by the change in active zone density during acute experimental manipulation or as a result of aging. Therefore, a mechanism must exist to maintain its density. In the central nervous system (CNS, active zones have restricted maximal size, exist in multiple numbers in larger presynaptic terminals, and maintain a constant density during development. These findings suggest that active zone density in the CNS is also controlled. However, in contrast to the NMJ, active zone density in the CNS can also be increased, as observed in hippocampal synapses in response to synaptic plasticity. Although the numbers of known active zone proteins and protein interactions have increased, less is known about the mechanism that controls the number or spacing of active zones. The following molecules are known to control active zone density and will be discussed herein: extracellular matrix laminins and voltage-dependent calcium channels, amyloid precursor proteins, the small GTPase Rab3, an endocytosis mechanism including synaptojanin, cytoskeleton protein spectrins and β-adducin, and a presynaptic web including spectrins. The molecular mechanisms that organize the active zone density are just beginning to be elucidated.

  3. Synaptic vesicle proteins and active zone plasticity

    Robert J Kittel

    2016-04-01

    Full Text Available Neurotransmitter is released from synaptic vesicles at the highly specialized presynaptic active zone. The complex molecular architecture of active zones mediates the speed, precision and plasticity of synaptic transmission. Importantly, structural and functional properties of active zones vary significantly, even for a given connection. Thus, there appear to be distinct active zone states, which fundamentally influence neuronal communication by controlling the positioning and release of synaptic vesicles. Vice versa, recent evidence has revealed that synaptic vesicle components also modulate organizational states of the active zone.The protein-rich cytomatrix at the active zone (CAZ provides a structural platform for molecular interactions guiding vesicle exocytosis. Studies in Drosophila have now demonstrated that the vesicle proteins Synaptotagmin-1 (Syt1 and Rab3 also regulate glutamate release by shaping differentiation of the CAZ ultrastructure. We review these unexpected findings and discuss mechanistic interpretations of the reciprocal relationship between synaptic vesicles and active zone states, which has heretofore received little attention.

  4. Optically active quantum-dot molecules.

    Shlykov, Alexander I; Baimuratov, Anvar S; Baranov, Alexander V; Fedorov, Anatoly V; Rukhlenko, Ivan D

    2017-02-20

    Chiral molecules made of coupled achiral semiconductor nanocrystals, also known as quantum dots, show great promise for photonic applications owing to their prospective uses as configurable building blocks for optically active structures, materials, and devices. Here we present a simple model of optically active quantum-dot molecules, in which each of the quantum dots is assigned a dipole moment associated with the fundamental interband transition between the size-quantized states of its confined charge carriers. This model is used to analytically calculate the rotatory strengths of optical transitions occurring upon the excitation of chiral dimers, trimers, and tetramers of general configurations. The rotatory strengths of such quantum-dot molecules are found to exceed the typical rotatory strengths of chiral molecules by five to six orders of magnitude. We also study how the optical activity of quantum-dot molecules shows up in their circular dichroism spectra when the energy gap between the molecular states is much smaller than the states' lifetime, and maximize the strengths of the circular dichroism peaks by optimizing orientations of the quantum dots in the molecules. Our analytical results provide clear design guidelines for quantum-dot molecules and can prove useful in engineering optically active quantum-dot supercrystals and photonic devices.

  5. Raman Optical Activity Spectra for Large Molecules through Molecules-in-Molecules Fragment-Based Approach.

    Jovan Jose, K V; Raghavachari, Krishnan

    2016-02-09

    We present an efficient method for the calculation of the Raman optical activity (ROA) spectra for large molecules through the molecules-in-molecules (MIM) fragment-based method. The relevant higher energy derivatives from smaller fragments are used to build the property tensors of the parent molecule to enable the extension of the MIM method for evaluating ROA spectra (MIM-ROA). Two factors were found to be particularly important in yielding accurate results. First, the link-atom tensor components are projected back onto the corresponding host and supporting atoms through the Jacobian projection method, yielding a mathematically rigorous method. Second, the long-range interactions between fragments are taken into account by using a less computationally expensive lower level of theory. The performance of the MIM-ROA model is calibrated on the enantiomeric pairs of 10 carbohydrate benchmark molecules, with strong intramolecular interactions. The vibrational frequencies and ROA intensities are accurately reproduced relative to the full, unfragmented, results for these systems. In addition, the MIM-ROA method is employed to predict the ROA spectra of d-maltose, α-D-cyclodextrin, and cryptophane-A, yielding spectra in excellent agreement with experiment. The accuracy and performance of the benchmark systems validate the MIM-ROA model for exploring ROA spectra of large molecules.

  6. The Proteome of the Murine Presynaptic Active Zone

    Melanie Laßek

    2014-04-01

    Full Text Available The proteome of the presynaptic active zone controls neurotransmitter release and the short- and long-term structural and functional dynamics of the nerve terminal. The proteinaceous inventory of the presynaptic active zone has recently been reported. This review will evaluate the subcellular fractionation protocols and the proteomic approaches employed. A breakthrough for the identification of the proteome of the presynaptic active zone was the successful employment of antibodies directed against a cytosolic epitope of membrane integral synaptic vesicle proteins for the immunopurification of synaptic vesicles docked to the presynaptic plasma membrane. Combining immunopurification and subsequent analytical mass spectrometry, hundreds of proteins, including synaptic vesicle proteins, components of the presynaptic fusion and retrieval machinery, proteins involved in intracellular and extracellular signaling and a large variety of adhesion molecules, were identified. Numerous proteins regulating the rearrangement of the cytoskeleton are indicative of the functional and structural dynamics of the presynapse. This review will critically discuss both the experimental approaches and prominent protein candidates identified. Many proteins have not previously been assigned to the presynaptic release sites and may be directly involved in the short- and long-term structural modulation of the presynaptic compartment. The identification of proteinaceous constituents of the presynaptic active zone provides the basis for further analyzing the interaction of presynaptic proteins with their targets and opens novel insights into the functional role of these proteins in neuronal communication.

  7. Synaptic Vesicle Proteins and Active Zone Plasticity.

    Kittel, Robert J; Heckmann, Manfred

    2016-01-01

    Neurotransmitter is released from synaptic vesicles at the highly specialized presynaptic active zone (AZ). The complex molecular architecture of AZs mediates the speed, precision and plasticity of synaptic transmission. Importantly, structural and functional properties of AZs vary significantly, even for a given connection. Thus, there appear to be distinct AZ states, which fundamentally influence neuronal communication by controlling the positioning and release of synaptic vesicles. Vice versa, recent evidence has revealed that synaptic vesicle components also modulate organizational states of the AZ. The protein-rich cytomatrix at the active zone (CAZ) provides a structural platform for molecular interactions guiding vesicle exocytosis. Studies in Drosophila have now demonstrated that the vesicle proteins Synaptotagmin-1 (Syt1) and Rab3 also regulate glutamate release by shaping differentiation of the CAZ ultrastructure. We review these unexpected findings and discuss mechanistic interpretations of the reciprocal relationship between synaptic vesicles and AZ states, which has heretofore received little attention.

  8. Photophysics of thermally activated delayed fluorescence molecules

    Dias, Fernando B.; Penfold, Thomas J.; Monkman, Andrew P.

    2017-03-01

    Thermally activated delayed fluorescence (TADF) has recently emerged as one of the most attractive methods for harvesting triplet states in metal-free organic materials for application in organic light emitting diodes (OLEDs). A large number of TADF molecules have been reported in the literature with the purpose of enhancing the efficiency of OLEDs by converting non-emissive triplet states into emissive singlet states. TADF emitters are able to harvest both singlets and triplet states through fluorescence (prompt and delayed), the latter due to the thermally activated reverse intersystem crossing mechanism that allows up-conversion of low energy triplet states to the emissive singlet level. This allows otherwise pure fluorescent OLEDs to overcome their intrinsic limit of 25% internal quantum efficiency (IQE), which is imposed by the 1:3 singlet–triplet ratio arising from the recombination of charges (electrons and holes). TADF based OLEDS with IQEs close to 100% are now routinely fabricated in the green spectral region. There is also significant progress for blue emitters. However, red emitters still show relatively low efficiencies. Despite the significant progress that has been made in recent years, still significant challenges persist to achieve full understanding of the TADF mechanism and improve the stability of these materials. These questions need to be solved in order to fully implement TADF in OLEDs and expand their application to other areas. To date, TADF has been exploited mainly in the field of OLEDs, but applications in other areas, such as sensing and fluorescence microscopies, are envisaged. In this review, the photophysics of TADF molecules is discussed, summarising current methods to characterise these materials and the current understanding of the TADF mechanism in various molecular systems.

  9. Active zone stability: insights from fly neuromuscular junction

    Xiaolin Tian

    2015-01-01

    Full Text Available The presynaptic active zone is a dynamic structure that orchestrates regulated release of neurotransmitters. Developmental and aging processes, and changes in neuronal network activity can all modulate the number, size and composition of active zone and thereby synaptic efficacy. However, very little is known about the mechanism that controls the structural stability of active zone. By studying a model synapse, the Drosophila neuromuscular junction, our recent work shed light on how two scaffolding proteins at the active zone regulate active zone stability by promoting a localized dephosphorylation event at the nerve terminal. Here we discuss the major insights from our findings and their implications for future research.

  10. Novel approaches for single molecule activation and detection

    Benfenati, Fabio; Torre, Vincent

    2014-01-01

    How can we obtain tools able to process and exchange information at the molecular scale In order to do this, it is necessary to activate and detect single molecules under controlled conditions. This book focuses on the generation of biologically-inspired molecular devices. These devices are based on the developments of new photonic tools able to activate and stimulate single molecule machines. Additionally, new light sensitive molecules can be selectively activated by photonic tools. These technological innovations will provide a way to control activation of single light-sensitive molecules, a

  11. Active zone stability:insights from fly neuromuscular junction

    Xiaolin Tian; Chunlai Wu

    2015-01-01

    The presynaptic active zone is a dynamic structure that orchestrates regulated release of neurotrans-mitters. Developmental and aging processes, and changes in neuronal network activity can all modulate the number, size and composition of active zone and thereby synaptic efifcacy. However, very little is known about the mechanism that controls the structural stability of active zone. By study-ing a model synapse, theDrosophila neuromuscular junction, our recent work shed light on how two scaffolding proteins at the active zone regulate active zone stability by promoting a localized dephos-phorylation event at the nerve terminal. Here we discuss the major insights from our ifndings and their implications for future research.

  12. Exploration of the spontaneous fluctuating activity of single enzyme molecules

    Schwabe, A.; Maarleveld, T.R.; Bruggeman, F.J.

    2013-01-01

    Single enzyme molecules display inevitable, stochastic fluctuations in their catalytic activity. In metabolism, for instance, the stochastic activity of individual enzymes is averaged out due to their high copy numbers per single cell. However, many processes inside cells rely on single enzyme activ

  13. Molecules and mechanisms that regulate multipolar migration in the intermediate zone.

    Cooper, Jonathan A

    2014-01-01

    Most neurons migrate with an elongated, "bipolar" morphology, extending a long leading process that explores the environment. However, when immature projection neurons enter the intermediate zone (IZ) of the neocortex they become "multipolar". Multipolar cells extend and retract cytoplasmic processes in different directions and move erratically-sideways, up and down. Multipolar cells extend axons while they are in the lower half of the IZ. Remarkably, the cells then resume radial migration: they reorient their centrosome and Golgi apparatus towards the pia, transform back to bipolar morphology, and commence locomotion along radial glia (RG) fibers. This reorientation implies the existence of directional signals in the IZ that are ignored during the multipolar stage but sensed after axonogenesis. In vivo genetic manipulation has implicated a variety of candidate directional signals, cell surface receptors, and signaling pathways, that may be involved in polarizing multipolar cells and stabilizing a pia-directed leading process for radial migration. Other signals are implicated in starting multipolar migration and triggering axon outgrowth. Here we review the molecules and mechanisms that regulate multipolar migration, and also discuss how multipolar migration affects the orderly arrangement of neurons in layers and columns in the developing neocortex.

  14. Ultrasensitive in situ visualization of active glucocerebrosidase molecules

    Witte, Martin D.; Kallemeijn, Wouter W.; Aten, Jan; Li, Kah-Yee; Strijland, Anneke; Donker-Koopman, Wilma E.; Nieuwendijk, Adrianus M.C.H. van den; Bleijlevens, Boris; Kramer, Gertjan; Florea, Bogdan I.; Hooibrink, Berend; Hollak, Carla E.M.; Ottenhoff, Roelof; Boot, Rolf G.; Marel, Gijsbert A. van der; Overkleeft, Herman S.; Aerts, Johannes M.F.G.

    2010-01-01

    Deficiency of glucocerebrosidase (GBA) underlies Gaucher disease, a common lysosomal storage disorder. Carriership for Gaucher disease has recently been identified as major risk for parkinsonism. Presently, no method exists to visualize active GBA molecules in situ. We here report the design, synthe

  15. Functionalization of hydroxyl terminated polybutadiene with biologically active fluorescent molecule

    R Murali Sankar; Subhadeep Saha; K Seeni Meera; Tushar Jana

    2009-10-01

    A biologically active molecule, 2-chloro-4,6-bis(dimethylamino)-1,3,5-triazine (CBDT), has been covalently attached at the terminal carbon atoms of the hydroxyl terminated polybutadiene (HTPB) backbone. The modification of HTPB backbone by CBDT molecule does not affect the unique physico-chemical properties such as fluidity, hydroxyl value and microstructure of the parent HTPB. The formation of hydrogen bonding between the terminal hydroxyl groups and the nitrogen atoms of triazine moiety is the driving force for the terminal attachment chemistry. The functionalized HTPB (HTPB–CBDT) shows a strong fluorescence emission at 385 nm.

  16. Decoupling Activation of Heme Biosynthesis from Anaerobic Toxicity in a Molecule Active in Staphylococcus aureus

    Dutter, Brendan F.; Mike, Laura A.; Reid, Paul R.; Chong, Katherine M.; Ramos-Hunter, Susan J.; Skaar, Eric P.; Sulikowski, Gary A.

    2016-01-01

    Small molecules active in the pathogenic bacterium Staphylococcus aureus are valuable tools for the study of its basic biology and pathogenesis, and many molecules may provide leads for novel therapeutics. We have previously reported a small molecule, 1, which activates endogenous heme biosynthesis in S. aureus, leading to an accumulation of intracellular heme. In addition to this novel activity, 1 also exhibits toxicity towards S. aureus growing under fermentative conditions. To determine if...

  17. Role of Adhesion Molecules in Eosinophil Activation: A Comparative Study on the Effect of Adhesion Molecules on Eosinophil Survival

    Kazutoshi Yamaguchi

    2004-01-01

    Conclusions: The regulation of adhesion molecules, by not only preventing eosinophil adhesion but also eosinophil activation, may be a potential target in the treatment of allergic inflammatory disorders.

  18. Structure and Structure-activity Relationship of Functional Organic Molecules

    2011-01-01

    @@ Research theme The group is made up of junior scientists from the State Key Laboratory of Elemento-organic Chemistry, Nankai University.The scientists focus their studis on the structure and structure-activity relationship of functional organic molecules not only because it has been the basis of their research, but also because the functional study of organic compounds is now a major scientific issue for organic chemists around the world.

  19. Polarized Raman optical activity of menthol and related molecules

    Barron, L. D.; Hecht, L.; Blyth, S. M.

    1989-01-01

    Polarized and depolarized Raman optical activity spectra of menthol, menthyl chloride, neomenthol and neothiomenthol from 800 to 1500 cm -1 are reported. Despite axial symmetry in all the bonds, the presence of the heteroatoms O or S seems to induce large deviations from the expected ratio of 2:1 between the polarized and depolarized Raman optical activity intensities, but Cl does not. These deviations might originate in large electric quadrupole contributions induced by excited state interactions involving O or S Rydberg p orbitals and valence orbitals on other parts of the molecule. Such interactions appear to undermine the bond polarizability theory of Raman intensities.

  20. Nano-sensing of the orientation of fluorescing molecules with active coated nano-particles

    Arslanagic, Samel; Ziolkowski, Richard W.

    2015-01-01

    The potential of using active coated nano-particles to determine the orientation of fluorescing molecules is reported. By treating each fluorescing molecule as an electric Hertzian dipole, single and multiple fluorescing molecules emitting coherently and incoherently in various orientations...

  1. Decoupling Activation of Heme Biosynthesis from Anaerobic Toxicity in a Molecule Active in Staphylococcus aureus.

    Dutter, Brendan F; Mike, Laura A; Reid, Paul R; Chong, Katherine M; Ramos-Hunter, Susan J; Skaar, Eric P; Sulikowski, Gary A

    2016-05-20

    Small molecules active in the pathogenic bacterium Staphylococcus aureus are valuable tools for the study of its basic biology and pathogenesis, and many molecules may provide leads for novel therapeutics. We have previously reported a small molecule, 1, which activates endogenous heme biosynthesis in S. aureus, leading to an accumulation of intracellular heme. In addition to this novel activity, 1 also exhibits toxicity towards S. aureus growing under fermentative conditions. To determine if these activities are linked and establish what features of the molecule are required for activity, we synthesized a library of analogs around the structure of 1 and screened them for activation of heme biosynthesis and anaerobic toxicity to investigate structure-activity relationships. The results of this analysis suggest that these activities are not linked. Furthermore, we have identified the structural features that promote each activity and have established two classes of molecules: activators of heme biosynthesis and inhibitors of anaerobic growth. These molecules will serve as useful probes for their respective activities without concern for the off target effects of the parent compound.

  2. Recent activity of Chihe segment of Tanlu fault zone

    姚大全; 刘加灿

    2004-01-01

    By means of differentiation of remote sensing image, field seismo-geological survey, analysis on drilling exploration materials, sampling and dating of rock samples, combined with seismicity and microscopic tectonic analysis, this paper studies the recent activity of Chihe segment of the Tanlu fault zone. The result indicates that the Chihe fault segment undergoes the deformation alternately in the mode of stick slip and creep during Late Quaternary, and its recent activity is mainly creep.

  3. Cannabinoids modulate Olig2 and polysialylated neural cell adhesion molecule expression in the subventricular zone of post-natal rats through cannabinoid receptor 1 and cannabinoid receptor 2.

    Arévalo-Martín, Angel; García-Ovejero, Daniel; Rubio-Araiz, Ana; Gómez, Oscar; Molina-Holgado, Francisco; Molina-Holgado, Eduardo

    2007-09-01

    The subventricular zone (SVZ) is a source of post-natal glial precursors that can migrate to the overlying white matter, where they may differentiate into oligodendrocytes. We showed that, in the post-natal SVZ ependymocytes, radial glia and astrocyte-like cells express cannabinoid receptor 1 (CB1), whereas cannabinoid receptor 2 (CB2) is found in cells expressing the polysialylated neural cell adhesion molecule. To study CB1 and CB2 function, post-natal rats were exposed to selective CB1 or CB2 agonists (arachidonyl-2-chloroethylamide and JWH-056, respectively) for 15 days. Accordingly, we found that CB1 activation increases the number of Olig2-positive cells in the dorsolateral SVZ, whereas CB2 activation increases polysialylated neural cell adhesion molecule expression in this region. As intense myelination occurs during the first weeks of post-natal development, we examined how modulating these factors affected the expression of myelin basic protein. Pharmacological administration of agonists and antagonists of CB1 and CB2 showed that the activation of both receptors is needed to augment the expression of myelin basic protein in the subcortical white matter.

  4. EVALUATION OF BIOLOGICALLY ACTIVE MOLECULES ISOLATED FROM OBLIGATE MARINE FUNGI

    A. KRISHNA SATYA

    2013-01-01

    Full Text Available The marine environment is a tremendous source of natural products. Marine microorganisms have become an important source of pharmacologically active metabolites Fungi are well known for their vast diversity of secondary metabolites that include many life-saving drugs and highly toxic mycotoxins. In general, fungal cultures producing such metabolites are immune to their toxic effects. However, some are known to produce self-toxic compounds that can pose production optimization challenges if the metabolites are needed in large amounts for chemical modification. Objective: The main objective of the present study was the isolation of new and preferably biologically active secondary metabolites from marine microorganisms, especially marine-derived fungi. Method: Marine fungi had isolated from marine soil by serial dilution method from Rose Bengal medium. Single colony was isolated by microscopic and macroscopic observation. Secondary metabolites are produced by marine fungi. Biological evaluation was performed by microbial studies. TLC is performed to identify the number of sub compounds in the crude extract. Further species level identification and structure elucidation of the compound are to be done. Results: The isolated marine fungi Aspergillus sp, showed maximum activity against the Candida rugosa with a zone diameter of 16mm at a concentration of 200μg and for bacterial strains it showed maximum activity against the E.coli with a diameter of 24mm at a concentration of 200μg. From the thin layer chromatography it has nearly 2-3 compounds to be purified. Conclusion: The selected organism which produces the compounds contains the biological activities which include anti-bacterial and anti fungal activities.

  5. Searching for line active molecules on biphasic lipid monolayers.

    Bischof, Andrea Alejandra; Mangiarotti, Agustín; Wilke, Natalia

    2015-03-21

    In membranes with phase coexistence, line tension appears as an important parameter for the determination of the amount of domains, as well as their size and their shape, thus defining the membrane texture. Different molecules have been proposed as "linactants" (i.e. molecules that reduce the line tension, thereby modulating the membrane texture). In this work, we explore the efficiency of different molecules as linactants in monolayers with two coexisting phases of different thicknesses. We tested the linactant ability of a molecule with chains of different saturation degrees, another molecule with different chain lengths and a bulky molecule. In this way, we show in the same system the effect of molecules with chains of different rigidities, with an intrinsic thickness mismatch and with a bulky moiety, thereby analyzing different hypotheses of how a molecule may change the line tension in a monolayer system. Both lipids with different hydrocarbon chains did not act as linactants, while only one of the bulky molecules tested decreased the line tension in the monolayer studied. We conclude that there are no universal rules for the structure of a molecule that enable us to predict that it will behave as a linactant and thus, designing linactants appears to be a difficult task and a challenge for future studies. Furthermore, in regard to the membrane texture, there was no direct influence of the line tension in the distribution of domain sizes.

  6. Bilirubin: an endogenous molecule with antiviral activity in vitro.

    Rosaria eSantangelo

    2012-03-01

    Full Text Available Bilirubin-IX-alpha (BR is the final product of heme metabolism through the heme oxygenase/biliverdin reductase (HO/BVR system. Previous papers reported on the microbicidal effects of the HO by-products biliverdin-IX-alpha, carbon monoxide and iron, through either direct or indirect mechanisms. In this paper the evidence of a virucidal effect of BR against human herpes simplex virus type 1 (HSV-1 and the enterovirus EV71 was provided. Bilirubin-IX-alpha, at concentrations 1-10 µM, close to those found in blood and tissues, significantly reduced HSV-1 and EV71 replication in Hep-2 and Vero cell lines, respectively. Bilirubin-IX-alpha inhibited viral infection of Hep-2 and Vero cells when given 2 hours before, concomitantly and 2 hours after viral infection. Furthermore, BR retained its antiviral activity even complexed with a saturating concentration of human serum-albumin. Moreover, 10 µM BR increased the formation of nitric oxide and the phosphorylation of JNK in Vero and Hep-2 cell lines, respectively, thus implying a role of these two pathways in the mechanism of antiviral activity of the bile pigment. In conclusion, these results support the antiviral effect of BR against HSV-1 and enterovirus in vitro, and put the basis for further basic and clinical studies to understand the real role of BR as an endogenous antiviral molecule.

  7. Signaling lymphocyte activating molecule (SLAM) expression in subacute sclerosing panencephalitis.

    Piskin, A Kevser; Akpinar, Pinar; Muftuoglu, Sevda; Anlar, Banu

    2007-08-01

    Signaling lymphocyte activating molecule (SLAM) is a receptor for measles virus which also has immunomodulatory activity. We analyzed SLAM expression in mononuclear cells (MNC) of patients with SSPE (n=7) and control subjects (n=7) from the same population. Native 10% PAGE analysis in cell and brain tissue extracts followed by Western blotting using monoclonal anti-human SLAM showed four types of bands. Differences in the type and amount of SLAM expression were observed between SSPE and control cases. Lymphocytes of SSPE patients showed two types of SLAM bands in comparison to only one in control lymphocytes. Stimulation of cells with lipopolysaccharide (80 u/ml) and concanavalin A (1 microg/ml) in vitro led to the appearance of a second isoform in both groups. Brain homogenates of SSPE patients (n=2) displayed all four types of SLAM isoforms at significantly higher levels than those of control brains (n=2). Our results show native PAGE enables the detection of all SLAM isotypes. The expression of SLAM is increased in lymphocytes, monocytes, and brain tissues of SSPE patients.

  8. The possibility of a fuzzy zone of semiotic activity.

    Morioka, Masayoshi

    2007-12-01

    In this commentary I tried to further develop the idea of Madureira (Integr Psych Behav Sci, 42(2), 2007), who challenges to clarify the complex sexuality problem of homophobia from the viewpoint of the cultural semiotic activity. Two remarking points were proposed in this commentary article. First, I took notice of the boundary phenomenon constructed between the homophobia and the other. It has a cultural meaning. Concerning with this process, I introduced and examined the concept of tonus that is sensed a subtle changing process of tension in self-other relationship. The second, I discussed about the fuzzy zone of semiotic activity. If one can feel in oneself fuzzy awareness into the source of discomfort affect, it is able to be a creative moment in the tension of fuzzy field (A and non-A) where generates dialogical activity in both vertical and horizontal. Through this discussion, I proposed some remarks for the dissolution on the culturally constructed prejudice of sexuality.

  9. Syntenin-1 and ezrin proteins link activated leukocyte cell adhesion molecule to the actin cytoskeleton

    Tudor, C.; Riet, J. te; Eich, C.; Harkes, R.; Smisdom, N.; Bouhuijzen-Wenger, J.; Ameloot, M.; Holt, M.; Kanger, J.S.; Figdor, C.G.; Cambi, A.; Subramaniam, V.

    2014-01-01

    Activated leukocyte cell adhesion molecule (ALCAM) is a type I transmembrane protein member of the immunoglobulin superfamily of cell adhesion molecules. Involved in important pathophysiological processes such as the immune response, cancer metastasis, and neuronal development, ALCAM undergoes both

  10. Migration and Extension of Solar Active Longitudinal Zones

    Gyenge, N; Ludmány, A

    2014-01-01

    Solar active longitudes show a characteristic migration pattern in the Carrington coordinate system when they can be identified at all. By following this migration, the longitudinal activity distribution around the center of the band can be determined. The halfwidth of the distribution is found to be varying in Cycles 21 - 23, and in some time intervals it was as narrow as 20 - 30 degrees. It was more extended around maximum but it was also narrow when the activity jumped to the opposite longitude. Flux emergence exhibited a quasi-periodic variation within the active zone with a period of about 1.3 years. The path of the active longitude migration does not support the view that it might be associated with the 11-year solar cycle. These results were obtained for a limited time interval of a few solar cycles and, bearing in mind uncertainties of the migration path definition, are only indicative. For the major fraction of dataset no systematic active longitudes were found. Sporadic migration of active longitude...

  11. Migration and Extension of Solar Active Longitudinal Zones

    Gyenge, N.; Baranyi, T.; Ludmány, A.

    2014-02-01

    Solar active longitudes show a characteristic migration pattern in the Carrington coordinate system if they can be identified at all. By following this migration, the longitudinal activity distribution around the center of the band can be determined. The half-width of the distribution is found to be varying in Cycles 21 - 23, and in some time intervals it was as narrow as 20 - 30 degrees. It was more extended around a maximum but it was also narrow when the activity jumped to the opposite longitude. Flux emergence exhibited a quasi-periodic variation within the active zone with a period of about 1.3 years. The path of the active-longitude migration does not support the view that it might be associated with the 11-year solar cycle. These results were obtained for a limited time interval of a few solar cycles and, bearing in mind uncertainties of the migration-path definition, are only indicative. For the major fraction of the dataset no systematic active longitudes were found. Sporadic migration of active longitudes was identified only for Cycles 21 - 22 in the northern hemisphere and Cycle 23 in the southern hemisphere.

  12. 78 FR 28801 - Foreign-Trade Zone 22-Chicago, Illinois; Authorization of Production Activity Panasonic...

    2013-05-16

    ... Foreign-Trade Zones Board Foreign-Trade Zone 22--Chicago, Illinois; Authorization of Production Activity Panasonic Corporation of North America (Kitting of Consumer Electronics) Aurora, Illinois On January 11... production activity to the Foreign-Trade Zones (FTZ) Board on behalf of Panasonic Corporation of...

  13. 78 FR 48647 - Foreign-Trade Zone 225-Springfield, Missouri; Authorization of Production Activity; General...

    2013-08-09

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF COMMERCE Foreign-Trade Zones Board Foreign-Trade Zone 225--Springfield, Missouri; Authorization of Production Activity; General Dynamics Ordnance and Tactical Systems Munitions Services (Demilitarization of...

  14. Super-resolution microscopy of the synaptic active zone.

    Ehmann, Nadine; Sauer, Markus; Kittel, Robert J

    2015-01-01

    Brain function relies on accurate information transfer at chemical synapses. At the presynaptic active zone (AZ) a variety of specialized proteins are assembled to complex architectures, which set the basis for speed, precision and plasticity of synaptic transmission. Calcium channels are pivotal for the initiation of excitation-secretion coupling and, correspondingly, capture a central position at the AZ. Combining quantitative functional studies with modeling approaches has provided predictions of channel properties, numbers and even positions on the nanometer scale. However, elucidating the nanoscopic organization of the surrounding protein network requires direct ultrastructural access. Without this information, knowledge of molecular synaptic structure-function relationships remains incomplete. Recently, super-resolution microscopy (SRM) techniques have begun to enter the neurosciences. These approaches combine high spatial resolution with the molecular specificity of fluorescence microscopy. Here, we discuss how SRM can be used to obtain information on the organization of AZ proteins.

  15. Super-resolution microscopy of the synaptic active zone

    Nadine eEhmann

    2015-01-01

    Full Text Available Brain function relies on accurate information transfer at chemical synapses. At the presynaptic active zone (AZ a variety of specialised proteins are assembled to complex architectures, which set the basis for speed, precision and plasticity of synaptic transmission.Calcium (Ca2+ channels are pivotal for the initiation of excitation-secretion coupling and, correspondingly, capture a central position at the AZ. Combining quantitative functional studies with modelling approaches has provided predictions of channel properties, numbers and even positions on the nanometre scale. However, elucidating the nanoscopic organisation of the surrounding protein network requires direct ultrastructural access. Without this information, knowledge of molecular synaptic structure-function relationships remains incomplete. Recently, super-resolution microscopy techniques have begun to enter the neurosciences. These approaches combine high spatial resolution with the molecular specificity of fluorescence microscopy. Here, we discuss how super-resolution microscopy can be used to obtain information on the organisation of AZ proteins.

  16. Quantitative super-resolution imaging of Bruchpilot distinguishes active zone states.

    Ehmann, Nadine; van de Linde, Sebastian; Alon, Amit; Ljaschenko, Dmitrij; Keung, Xi Zhen; Holm, Thorge; Rings, Annika; DiAntonio, Aaron; Hallermann, Stefan; Ashery, Uri; Heckmann, Manfred; Sauer, Markus; Kittel, Robert J

    2014-08-18

    The precise molecular architecture of synaptic active zones (AZs) gives rise to different structural and functional AZ states that fundamentally shape chemical neurotransmission. However, elucidating the nanoscopic protein arrangement at AZs is impeded by the diffraction-limited resolution of conventional light microscopy. Here we introduce new approaches to quantify endogenous protein organization at single-molecule resolution in situ with super-resolution imaging by direct stochastic optical reconstruction microscopy (dSTORM). Focusing on the Drosophila neuromuscular junction (NMJ), we find that the AZ cytomatrix (CAZ) is composed of units containing ~137 Bruchpilot (Brp) proteins, three quarters of which are organized into about 15 heptameric clusters. We test for a quantitative relationship between CAZ ultrastructure and neurotransmitter release properties by engaging Drosophila mutants and electrophysiology. Our results indicate that the precise nanoscopic organization of Brp distinguishes different physiological AZ states and link functional diversification to a heretofore unrecognized neuronal gradient of the CAZ ultrastructure.

  17. 78 FR 33808 - Foreign-Trade Zone 50-Long Beach, California; Authorization of Production Activity; Panasonic...

    2013-06-05

    ... No: 2013-13314] DEPARTMENT OF COMMERCE Foreign-Trade Zones Board [B-13-2013] Foreign-Trade Zone 50... (Kitting of Consumer Electronics); Anaheim, California On January 29, 2013, the Board of Harbor... activity to the Foreign-Trade Zones (FTZ) Board on behalf of Panasonic Corporation of North America,...

  18. Analysis of natural neutron flux in a seismically active zone

    V. F. Ostapenko

    2003-01-01

    Full Text Available In a seismically active zone in the near Almaty area (Kazakhstan since 1996 observations of variations of a natural neutron flux have been conducted. Sometimes the neutron flux rises sharply within the one-hour interval in comparison with the background. It occurs on the eve of activation of seismic processes. Increase of the neutron flux level had taken place from 1 h to 10 days prior to earthquakes. It is also indicated a tendency of growth of the anomaly level in accordance with the growth of energetic class of the subsequent earthquake. A character of connection between the neutron flux and earthquakes is still not clear. It is proposed that the neutron flux anomalies caused by variations of cosmic radiation intensity under action of fluxes of solar material, which is burst into interplanetary space (solar wind during solar flares. Energy of the solar wind transferred to Earth puts into action a trigger mechanism of the process of initiation of earthquakes at those places where conditions have already been prepared for them. The neutron flux anomalies can be used as substantial additional information for classical geophysical methods of short-term earthquake prediction.

  19. Structure-Activity Relation of AMOR Sugar Molecule That Activates Pollen-Tubes for Ovular Guidance.

    Jiao, Jiao; Mizukami, Akane G; Sankaranarayanan, Subramanian; Yamguchi, Junichiro; Itami, Kenichiro; Higashiyawma, Tetsuya

    2017-01-01

    Successful fertilization in flowering plants depends on the precise directional growth control of pollen tube through the female pistil tissue toward the female gametophyte contained in the ovule for delivery of nonmotile sperm cells. Cys-rich peptides LUREs secreted from the synergid cells on either side of the egg cell act as ovular attractants of pollen tubes. Competency control by the pistil is crucial for the response of pollen tubes to these ovular attractants. We recently reported that ovular 4-O-methyl-glucuronosyl arabinogalactan (AMOR) induces competency of the pollen tube to respond to ovular attractant LURE peptides in Torenia fournieri. The beta isomer of the terminal disaccharide 4-O-methyl-glucuronosyl galactose was essential and sufficient for the competency induction. However, critical and noncritical structures in the disaccharide have not been dissected deeply. Herein, we report the synthesis of new AMOR analogs and the structure-activity relationships for AMOR activity in the presence of these synthesized analogs. Removal of 4-O-methyl group or -COOH from the glucuronosyl residue of the disaccharide dramatically reduces AMOR activity. The pyranose backbone of the second sugar of disaccharide is essential for the activity but not hydroxy groups. The role of beta isomer of the disaccharide 4-Me-GlcA-β(1,6)-Gal is very specific for competency control, as there was no difference in effect among the sugar analogs tested for pollen germination. This study represents the first structure-activity relationship study, to our knowledge, of a sugar molecule involved in plant reproduction, which opens a way for modification of the molecule without loss of activity.

  20. Antimalarial Activity of Small-Molecule Benzothiazole Hydrazones

    Sarkar, Souvik; Siddiqui, Asim A.; Saha, Shubhra J.; De, Rudranil; Mazumder, Somnath; Banerjee, Chinmoy; Iqbal, Mohd S.; Nag, Shiladitya; Adhikari, Susanta

    2016-01-01

    We synthesized a new series of conjugated hydrazones that were found to be active against malaria parasite in vitro, as well as in vivo in a murine model. These hydrazones concentration-dependently chelated free iron and offered antimalarial activity. Upon screening of the synthesized hydrazones, compound 5f was found to be the most active iron chelator, as well as antiplasmodial. Compound 5f also interacted with free heme (KD [equilibrium dissociation constant] = 1.17 ± 0.8 μM), an iron-containing tetrapyrrole released after hemoglobin digestion by the parasite, and inhibited heme polymerization by parasite lysate. Structure-activity relationship studies indicated that a nitrogen- and sulfur-substituted five-membered aromatic ring present within the benzothiazole hydrazones might be responsible for their antimalarial activity. The dose-dependent antimalarial and heme polymerization inhibitory activities of the lead compound 5f were further validated by following [3H]hypoxanthine incorporation and hemozoin formation in parasite, respectively. It is worth mentioning that compound 5f exhibited antiplasmodial activity in vitro against a chloroquine/pyrimethamine-resistant strain of Plasmodium falciparum (K1). We also evaluated in vivo antimalarial activity of compound 5f in a murine model where a lethal multiple-drug-resistant strain of Plasmodium yoelii was used to infect Swiss albino mice. Compound 5f significantly suppressed the growth of parasite, and the infected mice experienced longer life spans upon treatment with this compound. During in vitro and in vivo toxicity assays, compound 5f showed minimal alteration in biochemical and hematological parameters compared to control. In conclusion, we identified a new class of hydrazone with therapeutic potential against malaria. PMID:27139466

  1. Exploring cellular memory molecules marking competent and active transcriptions

    Liu De-Pei

    2007-05-01

    Full Text Available Abstract Background Development in higher eukaryotes involves programmed gene expression. Cell type-specific gene expression is established during this process and is inherited in succeeding cell cycles. Higher eukaryotes have evolved elegant mechanisms by which committed gene-expression states are transmitted through numerous cell divisions. Previous studies have shown that both DNase I-sensitive sites and the basal transcription factor TFIID remain on silenced mitotic chromosomes, suggesting that certain trans-factors might act as bookmarks, maintaining the information and transmitting it to the next generation. Results We used the mouse globin gene clusters as a model system to examine the retention of active information on M-phase chromosomes and its contribution to the persistence of transcriptional competence of these gene clusters in murine erythroleukemia cells. In cells arrested in mitosis, the erythroid-specific activator NF-E2p45 remained associated with its binding sites on the globin gene loci, while the other major erythroid factor, GATA-1, was removed from chromosome. Moreover, despite mitotic chromatin condensation, the distant regulatory regions and promoters of transcriptionally competent globin gene loci are marked by a preserved histone code consisting in active histone modifications such as H3 acetylation, H3-K4 dimethylation and K79 dimethylation. Further analysis showed that other active genes are also locally marked by the preserved active histone code throughout mitotic inactivation of transcription. Conclusion Our results imply that certain kinds of specific protein factors and active histone modifications function as cellular memory markers for both competent and active genes during mitosis, and serve as a reactivated core for the resumption of transcription when the cells exit mitosis.

  2. Activated leukocyte cell adhesion molecule and prognosis in acute ischemic stroke

    Smedbakken, Linda; Jensen, Jesper K; Hallén, Jonas

    2011-01-01

    Biomarkers predicting mortality and functional outcome in stroke may be clinically helpful in identification of patients likely to benefit from intervention. Activated leukocyte cell adhesion molecule (ALCAM) is upregulated during neuroinflammation; we investigated whether ALCAM concentrations...

  3. Activation of MyD88 Signaling upon Staphylococcal Enterotoxin Binding to MHC Class II Molecules

    2011-01-20

    Activation of MyD88 Signaling upon Staphylococcal Enterotoxin Binding to MHC Class II Molecules Teri L. Kissner, Gordon Ruthel, Shahabuddin Alam...mediated signaling, which activates pro-inflammatory cytokine responses. Recently we reported that staphylococcal enterotoxins (SEA or SEB), which...upon Staphylococcal Enterotoxin Binding to MHC Class II Molecules. PLoS ONE 6(1): e15985. doi:10.1371/journal.pone.0015985 Editor: Jacques Zimmer

  4. Engineering of radiation of optically active molecules with chiral nano-meta-particles

    Klimov, Vasily V; Ducloy, Martial

    2011-01-01

    The radiation of optically active (chiral) molecule placed near chiral nanoparticle is investigated. The optimal conditions for engineering of radiation of optically active (chiral) molecules with the help of chiral nanoparticles are derived. It is shown that for this purpose, the substance of the chiral particle must have both \\epsilon_and \\mu_negative (double negative material (DNG)) or negative \\mu_and positive \\epsilon_(\\mu_negative material (MNG)). Our results pave the way to an effective engineering of radiation of "left" and "right" molecules and to creating pure optical devices for separation of drugs enantiomers.

  5. Engineering of radiation of optically active molecules with chiral nano-meta-particles

    Klimov, V. V.; Guzatov, D. V.; Ducloy, M.

    2012-02-01

    The radiation of an optically active (chiral) molecule placed near a chiral nanosphere is investigated. The optimal conditions for engineering of radiation of optically active (chiral) molecules with the help of chiral nanoparticles are derived. It is shown that for this purpose, the substance of the chiral particle must have both ɛ and μ negative (double negative material (DNG)) or negative μ and positive ɛ (μ negative material (MNG)). Our results pave the way to an effective engineering of radiation of "left" and "right" molecules and to creating pure optical devices for separation of drugs enantiomers.

  6. Release probability of hippocampal glutamatergic terminals scales with the size of the active zone.

    Holderith, Noemi; Lorincz, Andrea; Katona, Gergely; Rózsa, Balázs; Kulik, Akos; Watanabe, Masahiko; Nusser, Zoltan

    2012-06-10

    Cortical synapses have structural, molecular and functional heterogeneity; our knowledge regarding the relationship between their ultrastructural and functional parameters is still fragmented. Here we asked how the neurotransmitter release probability and presynaptic [Ca(2+)] transients relate to the ultrastructure of rat hippocampal glutamatergic axon terminals. Two-photon Ca(2+) imaging-derived optical quantal analysis and correlated electron microscopic reconstructions revealed a tight correlation between the release probability and the active-zone area. Peak amplitude of [Ca(2+)] transients in single boutons also positively correlated with the active-zone area. Freeze-fracture immunogold labeling revealed that the voltage-gated calcium channel subunit Cav2.1 and the presynaptic protein Rim1/2 are confined to the active zone and their numbers scale linearly with the active-zone area. Gold particles labeling Cav2.1 were nonrandomly distributed in the active zones. Our results demonstrate that the numbers of several active-zone proteins, including presynaptic calcium channels, as well as the number of docked vesicles and the release probability, scale linearly with the active-zone area.

  7. 78 FR 22843 - Foreign-Trade Zone 33-Pittsburgh, Pennsylvania, Authorization of Export Production Activity...

    2013-04-17

    ... Foreign-Trade Zones Board Foreign-Trade Zone 33--Pittsburgh, Pennsylvania, Authorization of Export... December 4, 2012, Tsudis Chocolate Company, submitted a notification of proposed export production activity... liquid chocolate admitted to FTZ 33 must be re-exported. Dated: April 11, 2013. Andrew...

  8. 77 FR 55455 - Foreign-Trade Zone 235-Lakewood, NJ, Authorization of Production Activity, Cosmetic Essence...

    2012-09-10

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF COMMERCE Foreign-Trade Zones Board Foreign-Trade Zone 235--Lakewood, NJ, Authorization of Production Activity, Cosmetic Essence Innovations, LLC, (Fragrance Bottling), Holmdel, NJ Cosmetic Essence Innovations, LLC...

  9. 78 FR 5773 - Foreign-Trade Zone 22-Chicago, IL, Notification of Proposed Production Activity, Panasonic...

    2013-01-28

    ... Foreign-Trade Zones Board Foreign-Trade Zone 22--Chicago, IL, Notification of Proposed Production Activity, Panasonic Corporation of North America, (Kitting of Consumer Electronics), Aurora, IL The Illinois... consumer electronics parts into retail packages. Pursuant to 15 CFR 400.14(b)4 of the regulations,...

  10. 78 FR 9667 - Foreign-Trade Zone 50-Long Beach, California; Notification of Proposed Production Activity...

    2013-02-11

    ... Foreign-Trade Zones Board Foreign-Trade Zone 50--Long Beach, California; Notification of Proposed Production Activity; Panasonic Corporation of North America (Kitting of Consumer Electronics); Anaheim, CA... FTZ 50. The facility is used for the kitting of consumer electronics parts into retail...

  11. 77 FR 70139 - Foreign-Trade Zone 8-Toledo, OH; Authorization of Production Activity; Whirlpool Corporation...

    2012-11-23

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF COMMERCE Foreign-Trade Zones Board Foreign-Trade Zone 8--Toledo, OH; Authorization of Production Activity; Whirlpool Corporation (Washing Machines); Clyde and Green Springs, OH On July 20, 2012 the...

  12. 77 FR 46024 - Foreign-Trade Zone 8-Toledo, OH; Notification of Proposed Production Activity, Whirlpool...

    2012-08-02

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF COMMERCE Foreign-Trade Zones Board Foreign-Trade Zone 8--Toledo, OH; Notification of Proposed Production Activity, Whirlpool Corporation (Washing Machines), Clyde and Green Springs, OH The Toledo-Lucas County Port...

  13. Ozone: A Multifaceted Molecule with Unexpected Therapeutic Activity.

    Zanardi, I; Borrelli, E; Valacchi, G; Travagli, V; Bocci, V

    2016-01-01

    A comprehensive outline for understanding and recommending the therapeutic use of ozone in combination with established therapy in diseases characterized by a chronic oxidative stress is currently available. The view of the absolute ozone toxicity is incorrect, because it has been based either on lung or on studies performed in artificial environments that do not correspond to the real antioxidant capacity of body compartments. In fact, ozone exerts either a potent toxic activity or it can stimulate biological responses of vital importance, analogously to gases with prospective therapeutic value such as NO, CO, H2S, H2, as well as O2 itself. Such a crucial difference has increasingly become evident during the last decade. The purpose of this review is to explain the aspects still poorly understood, highlighting the divergent activity of ozone on the various biological districts. It will be clarified that such a dual effect does not depend only upon the final gas concentration, but also on the particular biological system where ozone acts. The real significance of ozone as adjuvant therapeutic treatment concerns severe chronic pathologies among which are cardiovascular diseases, chronic obstructive pulmonary diseases, multiple sclerosis, and the dry form of age-related macular degeneration. It is time for a full insertion of ozone therapy within pharmaceutical sciences, responding to all the requirements of quality, efficacy and safety, rather than as either an alternative or an esoteric approach.

  14. Root Zone Microbial Populations, Urease Activities, and Purification Efficiency for a Constructed Wetland

    LIANG Wei; WU Zhen-Bin; ZHAN Fa-Cui; DENG Jia-Qi

    2004-01-01

    In order to investigate the effects of microorganisms and their urease activities in macrophytic root zones on pollutant removal, four small-scale plots (SSPs) of vertical/reverse-vertical flow wetlands were set up to determine: a) the relationship between the abundance of microorganisms in the root zones and water purification efficiency; and b) the relationship between urease activities in the root zones and pollutant removal in a constructed wetland system. Total numbers of the microbial population (bacteria, fungi, and actinomyces) along with urease activities in the macrophytic root zones were determined. In addition, the relationships between microbial populations and urease activities as well as the wastewater purification efficiencies of total phosphorus (TP), total Kjeldahl nitrogen (TKN), biochemical oxygen demand in 5 days (BOD5), and chemical oxygen demand (COD) were also analyzed. The results showed that there was a highly significant positive correlation (r = 0.9772, P < 0.01) between the number of bacteria in the root zones and BOD5 removal efficiency and a significant negative correlation (r = -0.9092, P < 0.05) between the number of fungi and the removal efficiency of TKN. Meanwhile, there was a significant positive correlation (r -- 0.8830, P < 0.05) between urease activities in the root zones and the removal efficiency of TKN. Thus, during wastewater treatment in a constructed wetland system,microorganism and urease activities in the root zones were very important factors.

  15. Associations between active living-oriented zoning and no adult leisure-time physical activity in the U.S.

    Leider, Julien; Chriqui, Jamie F; Thrun, Emily

    2017-02-01

    Nearly one-third of adults report no leisure-time physical activity (LTPA). Governmental and authoritative bodies recognize the role that community design through zoning code changes can play in enabling LTPA. This study examined the association between zoning and no adult LTPA in the U.S. This study was conducted between 2012 and 2016, with analyses occurring in 2015-2016. Zoning codes effective as of 2010 were compiled for jurisdictions located in the 495 most populous U.S. counties and were evaluated for pedestrian-oriented code reform zoning, 11 active living-oriented provisions (e.g., sidewalks, bike-pedestrian connectivity, mixed use, bike lanes) and a summated zoning scale (max=12). Individual-level LTPA data were obtained from the 2012 CDC Behavioral Risk Factor Surveillance System (BRFSS). County-aggregated, population-weighted zoning variables were constructed for linking to BRFSS. Log-log multivariate regressions (N=147,517 adults), controlling for individual and county characteristics and with robust standard errors clustered on county, were conducted to examine associations between zoning and no LTPA. Relative risks (RR) compared predicted lack of LTPA at 0% and 100% county-level population exposure to each zoning predictor. Zoning code reforms were associated with a 13% lower probability of no LTPA (RR: 0.87, 95% CI: 0.82-0.92). Except for crosswalks, all zoning provisions were associated with an 11-16% lower probability of no LTPA. Having all 12 zoning provisions was associated with a 22% lower probability of no LTPA (RR: 0.78, 95% CI: 0.72-0.83). The results suggest that active living-oriented zoning is a policy lever available to communities seeking to reduce rates of no LTPA.

  16. Exploring the Linkage between Activity-Friendly Zoning, Inactivity, and Cancer Incidence in the United States.

    Nicholson, Lisa M; Leider, Julien; Chriqui, Jamie F

    2017-03-07

    Background: Physical activity (PA) protects against cancer and enhances cancer survivorship. Given high inactivity rates nationwide, population-level physical activity facilitators are needed. Several authoritative bodies have recognized that zoning and planning helps create activity-friendly environments. This study examined the association between activity-friendly zoning, inactivity, and cancer in 478 of the most populous U.S. counties.Methods: County geocodes linked county-level data: cancer incidence and smoking (State Cancer Profiles), inactivity (Behavioral Risk Factor Surveillance System), 11 zoning measures (compiled by the study team), and covariates (from the American Community Survey and NAVTEQ). For each zoning measure, single mediation regression models and Sobel tests examined whether activity-friendly zoning was associated with reduced cancer incidence, and whether inactivity mediated those associations. All models were clustered on state with robust SEs and significance at the P Zoning for crosswalks, bike-pedestrian connectivity, and bike-pedestrian trails/paths were associated with reduced cancer incidence (β between -0.71 and -1.27, P zoning. Except for crosswalks, each association was mediated by inactivity. However, county smoking attenuated these results, with only crosswalks remaining significant. Results were similar for males (with zoning for bike-pedestrian connectivity, street connectivity, and bike-pedestrian trails/paths), but not females, alone.Conclusions: Zoning can help to create activity-friendly environments that support decreased inactivity, and possibly reduced cancer incidence.Impact: Given low physical activity levels nationwide, cross-sectoral collaborations with urban planning can inform cancer prevention and public health efforts to decrease inactivity and cancer. Cancer Epidemiol Biomarkers Prev; 1-9. ©2017 AACR.See all the articles in this CEBP Focus section, "Geospatial Approaches to Cancer Control and Population

  17. Investigating organic molecules responsible of auxin-like activity of humic acid fraction extracted from vermicompost

    Scaglia, Barbara, E-mail: barbara.scaglia@unimi.it [Gruppo Ricicla Labs – DiSAA, Università degli Studi di Milano, Via Celoria 2 (Italy); Nunes, Ramom Rachide; Rezende, Maria Olímpia Oliveira [Laboratório de Química Ambiental, Universidade de São Paulo, Instituto de Química de São Carlos, Avenida Trabalhador São Carlense, 400, São Carlos (Brazil); Tambone, Fulvia [Gruppo Ricicla Labs – DiSAA, Università degli Studi di Milano, Via Celoria 2 (Italy); Adani, Fabrizio, E-mail: fabrizio.adani@unimi.it [Gruppo Ricicla Labs – DiSAA, Università degli Studi di Milano, Via Celoria 2 (Italy)

    2016-08-15

    This work studied the auxin-like activity of humic acids (HA) obtained from vermicomposts produced using leather wastes plus cattle dung at different maturation stages (fresh, stable and mature). Bioassays were performed by testing HA concentrations in the range of 100–6000 mg carbon L{sup −1}. {sup 13}C CPMAS-NMR and GC–MS instrumental methods were used to assess the effect of biological processes and starting organic mixtures on HA composition. Not all HAs showed IAA-like activity and in general, IAA-like activity increased with the length of the vermicomposting process. The presence of leather wastes was not necessary to produce the auxin-like activity of HA, since HA extracted from a mix of cattle manure and sawdust, where no leather waste was added, showed IAA-like activity as well. CPMAS {sup 13}CNMR revealed that HAs were similar independently of the mix used and that the humification process involved the increasing concentration of pre-existing alkali soluble fractions in the biomass. GC/MS allowed the identification of the molecules involved in IAA-like effects: carboxylic acids and amino acids. The concentration of active molecules, rather than their simple presence in HA, determined the bio-stimulating effect, and a good linear regression between auxin-like activity and active stimulating molecules concentration was found (R{sup 2} = − 0.85; p < 0.01, n = 6). - Highlights: • Vermicomposting converts waste into organic fertilizer. • Vermicomposts can have biostimulating effect for the presence of hormone-like molecules. • Auxine-like activity was associated to the vermicompost humic acid fraction (HA). • HA carboxylic acids and amino acids, were reported to act as auxin-like molecules. • A linear regression was found between molecules and auxin-like activity.

  18. [Differences of activations in visual and associative zones during figurative and verbal activity].

    Nagornova, Zh V; Shemiakina, N V

    2014-04-01

    The study considers correlates of figurative and verbal tasks performance during attention paid to visual stimuli. There are 34 subjects (20 female, mean age 21, 2.5 [SD]) took parts in the study. During subjects performance of the task, there was carried out EEG registration from 19 sites according to 10-20%. Performance of the figurative creative task in comparison with control non-creative task of the same modality was accompanied by activation of occipital and parietal zones of the cerebral cortex (decrease of EEG spectral power in alpha 1 (7.5-9.5 Hz) and alpha2 (10-12.5 Hz) frequency bands was observed) whereas performance of a verbal creative task in the similar test-control comparison was accompanied by decrease of activation in occipital zones (revealed through increase of EEG spectral in alphal and alpha2 frequency bands). As visual stimuli were shown during the whole time of the creative and control tasks fulfilment was made an assumption observed distinction can be connected with redistribution of attention focus at various types of creative activity (figurative or verbal).

  19. Expression of adhesion and activation molecules on lymphocytes during open-heart surgery with cardiopulmonary bypass

    Toft, P; Tønnesen, Else Kirstine; Zülow, I

    1997-01-01

    Open-heart surgery with cardiopulmonary bypass (CPB) and abdominal surgery are associated with lymphocytopenia. We measured a panel of adhesion and activation molecules on lymphocytes to clarify possible association of CPB with increased expression of these molecules. Eight patients undergoing open......-heart surgery and eight with abdominal surgery were studied. The adhesion molecules CD11a/CD18 (LFA-1_, CD11c/CD18 and CD44 and the activation molecules CD25, CD69, CD71 and MHCII were measured, using monoclonal antibodies and flow cytometry. Lymphocytopenia was observed during CPB and for some hours after both...... open-heart and abdominal surgery. The proportion of CD11a/CD18-positive lymphocytes rose from 67.6 +/- 8% to 86.4 +/- 3% after aortic declamping (p

  20. Presynaptic spinophilin tunes neurexin signalling to control active zone architecture and function

    Muhammad, Karzan; Reddy-Alla, Suneel; Driller, Jan H; Schreiner, Dietmar; Rey, Ulises; Böhme, Mathias A.; Hollmann, Christina; Ramesh, Niraja; Depner, Harald; Lützkendorf, Janine; Matkovic, Tanja; Götz, Torsten; Bergeron, Dominique D.; Schmoranzer, Jan; Goettfert, Fabian; Holt, Mathew; Wahl, Markus C.; Hell, Stefan W.; Scheiffele, Peter; Walter, Alexander M.; Loll, Bernhard; Sigrist, Stephan J.

    2015-01-01

    Assembly and maturation of synapses at the Drosophila neuromuscular junction (NMJ) depend on trans-synaptic neurexin/neuroligin signalling, which is promoted by the scaffolding protein Syd-1 binding to neurexin. Here we report that the scaffold protein spinophilin binds to the C-terminal portion of neurexin and is needed to limit neurexin/neuroligin signalling by acting antagonistic to Syd-1. Loss of presynaptic spinophilin results in the formation of excess, but atypically small active zones. Neuroligin-1/neurexin-1/Syd-1 levels are increased at spinophilin mutant NMJs, and removal of single copies of the neurexin-1, Syd-1 or neuroligin-1 genes suppresses the spinophilin-active zone phenotype. Evoked transmission is strongly reduced at spinophilin terminals, owing to a severely reduced release probability at individual active zones. We conclude that presynaptic spinophilin fine-tunes neurexin/neuroligin signalling to control active zone number and functionality, thereby optimizing them for action potential-induced exocytosis. PMID:26471740

  1. Auroral Electrojet Index Designed to Provide a Global Measure, Hourly Intervals, of Auroral Zone Magnetic Activity

    National Oceanic and Atmospheric Administration, Department of Commerce — The Auroral Electrojet (AE) index is designed to provide a global quantitative measure of auroral zone magnetic activity produced by enhanced ionospheric currents...

  2. Potato signal molecules that activate pectate lyase synthesis in Pectobacterium atrosepticum SCRI1043.

    Tarasova, Nadezhda; Gorshkov, Vladimir; Petrova, Olga; Gogolev, Yuri

    2013-07-01

    A new type of plant-derived signal molecules that activate extracellular pectate lyase activity in phytopathogenic bacterium Pectobacterium atrosepticum SCRI1043 was revealed. These compounds were characterized and partially purified by means of several approaches including RT-PCR analysis, luminescence bioassay and HPLC fractionation. They were smaller than 1 kDa, thermoresistant, nonproteinaceous, hydrophilic, and slightly negatively charged molecules. Using gene expression analysis and bacterial biosensor assay the mode of activity of revealed compounds was studied. The possibility of their action through quorum sensing- and KdgR-mediated pathways was analyzed.

  3. Active fault survey on the Tanlu fault zone in Laizhou Bay

    WANG Zhi-cai; YANG Xi-ha; LI Chang-chuan; DENG Qi-dong; DU Xian-song; CHAO Hong-tai; WU Zi-quan; XIAO Lan-xi; SUN Zhao-ming; MIN Wei; LING Hong

    2006-01-01

    Shallow-depth acoustic reflection profiling survey has been conducted on the Tanlu fault zone in Laizhou Bay. It is found that the Tanlu fault zone is obviously active during the late Quaternary and it is still the dominating structure in this region. The Tanlu fault zone consists of two branches. The KL3 fault of the western branch is composed of several high angle normal faults which had been active during the period from the latest Pleistocene to early Holocene, dissected by a series of northeast or approximate east-west trending fault which leaped sediment of the late Pleistocene. The Longkou fault of the eastern branch consists of two right-laterally stepped segments. Late Quaternary offsets and growth strata developed along the Tanlu fault zone verify that the fault zone retained active in the latest Pleistocene to the early Holocene. The Anqiu-Juxian fault that passes through the middle of Shandong and corresponds to the Longkou fault is composed of a series of right-laterally stepped segments. The active faults along the eastern branch of the Tanlu fault zone from the Laizhou bay to the north of Anqiu make up a dextral simple shear deformation zone which is characterized by right-lateral strike-slip movement with dip-slip component during the late Quaternary.

  4. Group Problem Solving as a Zone of Proximal Development activity

    Brewe, Eric

    2006-12-01

    Vygotsky described learning as a process, intertwined with development, which is strongly influenced by social interactions with others that are at differing developmental stages.i These interactions create a Zone of Proximal Development for each member of the interaction. Vygotsky’s notion of social constructivism is not only a theory of learning, but also of development. While teaching introductory physics in an interactive format, I have found manifestations of Vygotsky’s theory in my classroom. The source of evidence is a paired problem solution. A standard mechanics problem was solved by students in two classes as a homework assignment. Students handed in the homework and then solved the same problem in small groups. The solutions to both the group and individual problem were assessed by multiple reviewers. In many cases the group score was the same as the highest individual score in the group, but in some cases, the group score was higher than any individual score. For this poster, I will analyze the individual and group scores and focus on three groups solutions and video that provide evidence of learning through membership in a Zone of Proximal Development. Endnotes i L. Vygotsky -Mind and society: The development of higher mental processes. Cambridge, MA: Harvard University Press. (1978).

  5. Activation of complement by an IgG molecule without a genetic hinge.

    Brekke, O H; Michaelsen, T E; Sandin, R; Sandlie, I

    1993-06-17

    The hinge region links the two Fab arms to the Fc portion of the IgG molecule. It mediates flexibility to the molecule and serves as a connecting structure between the two heavy chains. In addition it provides space between the Fab and Fc parts. All three properties have been proposed to be important for the ability of IgG to initiate complement activation leading to complement-mediated cell lysis (CML). Here we report the construction of a hinge-deleted mouse-human chimaeric IgG3 molecule with specificity for the hapten NIP (3-iodo-4-hydroxy-5-nitrophenacetyl), HM-1. HM-1 lacks the genetic hinge, but has an introduced cysteine between Ala 231 (EU numbering) and Pro 232 in the lower hinge encoded by the CH2 exon. The introduced cysteine forms a disulphide bond between the two heavy chains of the molecule. In CML, HM-1 shows a greater activity than IgG3 wild type. This is the first time an IgG molecule without a genetic hinge has been found to be active in CML. We conclude that the hinge functioning as a spacer is not a prerequisite for complement activation. Rather, its major role seems to be to connect the heavy chains to each other in the amino-terminal part of CH2. Because HM-1 is expected to have low Fab-Fc flexibility, this molecular feature is probably of no importance for complement activation.

  6. Line active molecules promote inhomogeneous structures in membranes: theory, simulations and experiments.

    Palmieri, Benoit; Yamamoto, Tetsuya; Brewster, Robert C; Safran, Samuel A

    2014-06-01

    We review recent theoretical efforts that predict how line-active molecules can promote lateral heterogeneities (or domains) in model membranes. This fundamental understanding may be relevant to membrane composition in living cells, where it is thought that small domains, called lipid rafts, are necessary for the cells to be functional. The theoretical work reviewed here ranges in scale from coarse grained continuum models to nearly atomistic models. The effect of line active molecules on domain sizes and shapes in the phase separated regime or on fluctuation length scales and lifetimes in the single phase, mixed regime, of the membrane is discussed. Recent experimental studies on model membranes that include line active molecules are also presented together with some comparisons with the theoretical predictions.

  7. Photo-activation of Single Molecule Magnet Behavior in a Manganese-based Complex

    Fetoh, Ahmed; Cosquer, Goulven; Morimoto, Masakazu; Irie, Masahiro; El-Gammal, Ola; El-Reash, Gaber Abu; Breedlove, Brian K.; Yamashita, Masahiro

    2016-03-01

    A major roadblock to fully realizing molecular electronic devices is the ability to control the properties of each molecule in the device. Herein we report the control of the magnetic properties of single-molecule magnets (SMMs), which can be used in memory devices, by using a photo-isomerizable diarthylenthene ligand. Photo-isomerization of the diarylethene ligand bridging two manganese salen complexes with visible light caused a significant change in the SMM behavior due to opening of the six-membered ring of diarylethene ligand, accompanied by reorganization of the entire molecule. The ring-opening activated the frequency-dependent magnetization of the complex. Our results are a major step towards the realization of molecular memory devices composed of SMMs because the SMM behaviour can be turned on and off simply by irradiating the molecule.

  8. Implication of Crystal Water Molecules in Inhibitor Binding at ALR2 Active Site

    Hymavati

    2012-01-01

    Full Text Available Water molecules play a crucial role in mediating the interaction between a ligand and a macromolecule. The solvent environment around such biomolecule controls their structure and plays important role in protein-ligand interactions. An understanding of the nature and role of these water molecules in the active site of a protein could greatly increase the efficiency of rational drug design approaches. We have performed the comparative crystal structure analysis of aldose reductase to understand the role of crystal water in protein-ligand interaction. Molecular dynamics simulation has shown the versatile nature of water molecules in bridge H bonding during interaction. Occupancy and life time of water molecules depend on the type of cocrystallized ligand present in the structure. The information may be useful in rational approach to customize the ligand, and thereby longer occupancy and life time for bridge H-bonding.

  9. In Vitro Selection for Small-Molecule-Triggered Strand Displacement and Riboswitch Activity.

    Martini, Laura; Meyer, Adam J; Ellefson, Jared W; Milligan, John N; Forlin, Michele; Ellington, Andrew D; Mansy, Sheref S

    2015-10-16

    An in vitro selection method for ligand-responsive RNA sensors was developed that exploited strand displacement reactions. The RNA library was based on the thiamine pyrophosphate (TPP) riboswitch, and RNA sequences capable of hybridizing to a target duplex DNA in a TPP regulated manner were identified. After three rounds of selection, RNA molecules that mediated a strand exchange reaction upon TPP binding were enriched. The enriched sequences also showed riboswitch activity. Our results demonstrated that small-molecule-responsive nucleic acid sensors can be selected to control the activity of target nucleic acid circuitry.

  10. Is the effect of surface modifying molecules on antibacterial activity universal for a given material?

    Hsu, Alexander; Liu, Fangzhou; Leung, Yu Hang; Ma, Angel P. Y.; Djurišić, Aleksandra B.; Leung, Frederick C. C.; Chan, Wai Kin; Lee, Hung Kay

    2014-08-01

    Antibacterial activity of nanomaterials is strongly dependent on their properties, and their stability and toxicity can be varied using surface coatings. We investigated the effect of different surface modifying molecules on the antibacterial properties of two ZnO nanoparticle samples. We found that the starting surface properties of the nanoparticles have significant effects on the attachment of the surface modifying molecules and consequent antibacterial activity. Two out of five investigated surface modifying molecules not only had a significant difference in the magnitude of their effect on different nanoparticles, but also resulted in the opposite effects on two ZnO nanoparticle samples (an enhancement of antibacterial activity for one and a reduction of antibacterial activity for the other ZnO sample). This indicates that no general rule on the effect of a specific molecule on the toxicity of a metal oxide nanoparticle can be derived without knowing the nanoparticle properties, due to the fact that surface modifier attachment onto the surface is affected by the initial surface properties.Antibacterial activity of nanomaterials is strongly dependent on their properties, and their stability and toxicity can be varied using surface coatings. We investigated the effect of different surface modifying molecules on the antibacterial properties of two ZnO nanoparticle samples. We found that the starting surface properties of the nanoparticles have significant effects on the attachment of the surface modifying molecules and consequent antibacterial activity. Two out of five investigated surface modifying molecules not only had a significant difference in the magnitude of their effect on different nanoparticles, but also resulted in the opposite effects on two ZnO nanoparticle samples (an enhancement of antibacterial activity for one and a reduction of antibacterial activity for the other ZnO sample). This indicates that no general rule on the effect of a specific

  11. 34 CFR 299.3 - What priority may the Secretary establish for activities in an Empowerment Zone or Enterprise...

    2010-07-01

    ... activities in an Empowerment Zone or Enterprise Community? 299.3 Section 299.3 Education Regulations of the... activities in an Empowerment Zone or Enterprise Community? For any ESEA discretionary grant program, the... significant portion of the program funds to address substantial problems in an Empowerment Zone, including...

  12. Brazing zone structure at active brazing of alumina ceramics

    2005-01-01

    @@ Nowadays one of the most effective methods of joining of oxide ceramics with other elements of construction is active brazing based on using of active metals (Ti, Zr), which increase reactivity of brazing alloy relative to ceramic element of a joining.

  13. Brazing zone structure at active brazing of alumina ceramics

    Demchuk; V.; A.; Kalinichenko; B.; B.

    2005-01-01

    Nowadays one of the most effective methods of joining of oxide ceramics with other elements of construction is active brazing based on using of active metals (Ti, Zr), which increase reactivity of brazing alloy relative to ceramic element of a joining.……

  14. Approach for targeting Ras with small molecules that activate SOS-mediated nucleotide exchange.

    Burns, Michael C; Sun, Qi; Daniels, R Nathan; Camper, DeMarco; Kennedy, J Phillip; Phan, Jason; Olejniczak, Edward T; Lee, Taekyu; Waterson, Alex G; Rossanese, Olivia W; Fesik, Stephen W

    2014-03-01

    Aberrant activation of the small GTPase Ras by oncogenic mutation or constitutively active upstream receptor tyrosine kinases results in the deregulation of cellular signals governing growth and survival in ∼30% of all human cancers. However, the discovery of potent inhibitors of Ras has been difficult to achieve. Here, we report the identification of small molecules that bind to a unique pocket on the Ras:Son of Sevenless (SOS):Ras complex, increase the rate of SOS-catalyzed nucleotide exchange in vitro, and modulate Ras signaling pathways in cells. X-ray crystallography of Ras:SOS:Ras in complex with these molecules reveals that the compounds bind in a hydrophobic pocket in the CDC25 domain of SOS adjacent to the Switch II region of Ras. The structure-activity relationships exhibited by these compounds can be rationalized on the basis of multiple X-ray cocrystal structures. Mutational analyses confirmed the functional relevance of this binding site and showed it to be essential for compound activity. These molecules increase Ras-GTP levels and disrupt MAPK and PI3K signaling in cells at low micromolar concentrations. These small molecules represent tools to study the acute activation of Ras and highlight a pocket on SOS that may be exploited to modulate Ras signaling.

  15. Dynamics of water molecules in the active-site cavity of human cytochromes P450

    Rydberg, Patrik; Rod, Thomas Holm; Olsen, Lars;

    2007-01-01

    have quite big cavities, with 41 water molecules on average in 2C8 and 54-58 in 2C9 and 3A4, giving a water volume of 1500-2100 A3. The two crystal structures of 2C9 differ quite appreciably, whereas those of 3A4 are quite similar. The active-site cavity is connected to the surroundings by three to six......We have studied the dynamics of water molecules in six crystal structures of four human cytochromes P450, 2A6, 2C8, 2C9, and 3A4, with molecular dynamics simulations. In the crystal structures, only a few water molecules are seen and the reported sizes of the active-site cavity vary a lot...

  16. Novel Molecular Non-Volatile Memory: Application of Redox-Active Molecules

    Hao Zhu

    2015-12-01

    Full Text Available This review briefly describes the development of molecular electronics in the application of non-volatile memory. Molecules, especially redox-active molecules, have become interesting due to their intrinsic redox behavior, which provides an excellent basis for low-power, high-density and high-reliability non-volatile memory applications. Recently, solid-state non-volatile memory devices based on redox-active molecules have been reported, exhibiting fast speed, low operation voltage, excellent endurance and multi-bit storage, outperforming the conventional floating-gate flash memory. Such high performance molecular memory will lead to promising on-chip memory and future portable/wearable electronics applications.

  17. Investigating organic molecules responsible of auxin-like activity of humic acid fraction extracted from vermicompost.

    Scaglia, Barbara; Nunes, Ramom Rachide; Rezende, Maria Olímpia Oliveira; Tambone, Fulvia; Adani, Fabrizio

    2016-08-15

    This work studied the auxin-like activity of humic acids (HA) obtained from vermicomposts produced using leather wastes plus cattle dung at different maturation stages (fresh, stable and mature). Bioassays were performed by testing HA concentrations in the range of 100-6000mgcarbonL(-1). (13)C CPMAS-NMR and GC-MS instrumental methods were used to assess the effect of biological processes and starting organic mixtures on HA composition. Not all HAs showed IAA-like activity and in general, IAA-like activity increased with the length of the vermicomposting process. The presence of leather wastes was not necessary to produce the auxin-like activity of HA, since HA extracted from a mix of cattle manure and sawdust, where no leather waste was added, showed IAA-like activity as well. CPMAS (13)CNMR revealed that HAs were similar independently of the mix used and that the humification process involved the increasing concentration of pre-existing alkali soluble fractions in the biomass. GC/MS allowed the identification of the molecules involved in IAA-like effects: carboxylic acids and amino acids. The concentration of active molecules, rather than their simple presence in HA, determined the bio-stimulating effect, and a good linear regression between auxin-like activity and active stimulating molecules concentration was found (R(2)=-0.85; p<0.01, n=6).

  18. Role of signaling lymphocytic activation molecule in T helper cell responses

    Jan E. de Vries

    1998-01-01

    Full Text Available Signaling lymphocytic activation molecule (SLAM; CDw150 is a 70 kDa glycoprotein. Signaling lymphocytic activation molecule is constitutively expressed on memory T cells, CD56+ T cells, a subset of T cell receptor γδ+ cells, immature thymocytes and, at low levels, on a proportion of peripheral blood B cells. Signaling lymphocytic activation molecule is rapidly upregulated on all T and B cells after activation. Engagement of SLAM by F(ab’2 fragments of an anti-SLAM monoclonal antibody (mAb A12 enhances antigen-specific T cell proliferation. In addition, mAb A12 was directly mitogenic for T cell clones and activated T cells. T cell proliferation induced by mAb A12 is independent of interleukin (IL-2, IL-4, IL-12 and IL-15, but is cyclosporin A sensitive. Ligation of SLAM during antigen-specific T cell proliferation resulted in upregulation of interferon (IFN-γ production, even by allergen-specific T helper cell (Th 2 clones, whereas the levels of IL-4 and IL-5 production were only marginally affected. The mAb A12 was unable to induce IL-4 and IL-5 production by Th1 clones. Co-stimulation of skin-derived Der P1-specific Th2 cells from patients with atopic dermatitis via SLAM resulted in the generation of a population of IFN-γ-producing cells, thereby reverting their phenotype to a Th0 pattern. Signaling lymphocytic activation molecule is a high-affinity self ligand mediating homophilic cell interaction. In addition, soluble SLAM enhances both T and B cell proliferation. Collectively, these data indicate that SLAM molecules act both as receptors and ligands that are not only involved in T cell expansion but also drive the expanding T cells during immune responses into the Th0/Th1 pathway. This suggests that signaling through SLAM plays a role in directing Th0/Th1 development.

  19. Single-active-electron potentials for molecules in intense laser fields

    Abu-Samha, Mahmoud; Madsen, Lars Bojer

    2010-01-01

    Single-active-electron potentials are computed for selected molecules, and molecular wave functions with the correct asymptotic behavior are produced. Asymptotic expansion coefficients are extracted from the wave functions and used to compute alignment-dependent ionization yields from molecular...

  20. Bio-active molecules modified surfaces enhanced mesenchymal stem cell adhesion and proliferation.

    Mobasseri, Rezvan; Tian, Lingling; Soleimani, Masoud; Ramakrishna, Seeram; Naderi-Manesh, Hossein

    2017-01-29

    Surface modification of the substrate as a component of in vitro cell culture and tissue engineering, using bio-active molecules including extracellular matrix (ECM) proteins or peptides derived ECM proteins can modulate the surface properties and thereby induce the desired signaling pathways in cells. The aim of this study was to evaluate the behavior of human bone marrow mesenchymal stem cells (hBM-MSCs) on glass substrates modified with fibronectin (Fn), collagen (Coll), RGD peptides (RGD) and designed peptide (R-pept) as bio-active molecules. The glass coverslips were coated with fibronectin, collagen, RGD peptide and R-peptide. Bone marrow mesenchymal stem cells were cultured on different substrates and the adhesion behavior in early incubation times was investigated using scanning electron microscopy (SEM) and confocal microscopy. The MTT assay was performed to evaluate the effect of different bio-active molecules on MSCs proliferation rate during 24 and 72 h. Formation of filopodia and focal adhesion (FA) complexes, two steps of cell adhesion process, were observed in MSCs cultured on bio-active molecules modified coverslips, specifically in Fn coated and R-pept coated groups. SEM image showed well adhesion pattern for MSCs cultured on Fn and R-pept after 2 h incubation, while the shape of cells cultured on Coll and RGD substrates indicated that they might experience stress condition in early hours of culture. Investigation of adhesion behavior, as well as proliferation pattern, suggests R-peptide as a promising bio-active molecule to be used for surface modification of substrate in supporting and inducing cell adhesion and proliferation.

  1. Spin state transition in the active center of the hemoglobin molecule: DFT + DMFT study

    Novoselov, D.; Korotin, Dm. M.; Anisimov, V. I.

    2016-05-01

    An ab initio study of electronic and spin configurations of the iron ion in the active center of the human hemoglobin molecule is presented. With a combination of the Density Functional Theory (DFT) method and the Dynamical Mean Field Theory (DMFT) approach, the spin state transition description in the iron ion during the oxidation process is significantly improved in comparison with previous attempts. It was found that the origin of the iron ion local moment behavior both for the high-spin and for the low-spin states in the hemoglobin molecule is caused by the presence of a mixture of several atomic states with comparable statistical probability.

  2. Heavy metals contamination characteristics in soil of different mining activity zones

    LIAO Guo-li; LIAO Da-xue; LI Quan-ming

    2008-01-01

    Depending upon the polluted features of various mining activities in a typical nonferrous metal mine, the contaminated soil area was divided into four zones which were polluted by tailings, mine drainage, dust deposition in wind and spreading minerals during vehicle transportation, respectively. In each zone, soil samples were collected. Total 28 soil samples were dug and analyzed by ICP-AES and other relevant methods. The results indicate that the average contents of Zn, Pb, Cd, Cu and As in soils are 508.6, 384.8, 7.53, 356 and 44.6 mg/kg, respectively. But the contents of heavy metals in different zone have distinct differences. The proportion of oxidizing association with organic substance is small. Difference of the association of heavy metals is small in different polluted zones.

  3. The Ms7.0 Lushan earthquake and the activity of the Longmenshan fault zone

    Meng Xiangang

    2013-08-01

    Full Text Available The Ms7.0 Lushan earthquake is directly related to the activity of Longmenshan fault zone. In this article, deformation monitoring data in Longmenshan and its surrounding areas were analyzed and the result shows that the activity trend of Longmenshan fault zone depends on the relative motion between Bayan Har Block and Sichuan Basin, and the main power of the movement comes from the Tibetan Plateau and the upper Yangtze craton massif of push. In recent years, the Longmenshan and its surrounding areas is one of the main seismogenic area in mainland China. In this paper, combination with seismogenic area of geological structure and crustal deformation observation data analysis results, the relationship between the earthquake and Longmenshan fault zone activity was discussed, and the key monitoring areas in the next five years were proposed.

  4. Single molecule investigation of the onset and minimum size of the calcium-mediated junction zone in alginate.

    Bowman, Kate A; Aarstad, Olav Andreas; Nakamura, Marcela; Stokke, Bjørn Torger; Skjåk-Bræk, Gudmund; Round, Andrew N

    2016-09-05

    One of the principal roles of alginate, both natively and in commercial applications, is gelation via Ca(2+)-mediated crosslinks between blocks of guluronic acid. In this work, single molecule measurements were carried out between well-characterised series of nearly monodisperse guluronic acid blocks ('oligoGs') using dynamic force spectroscopy. The measurements provide evidence that for interaction times on the order of tens of milliseconds the maximum crosslink strength is achieved by pairs of oligoGs long enough to allow the coordination of 4Ca(2+) ions, with both shorter and longer oligomers forming weaker links. Extending the interaction time from tens to hundreds of milliseconds allows longer oligoGs to achieve much stronger crosslinks but does not change the strength of individual links between shorter oligoGs. These results are considered in light of extant models for the onset of cooperative crosslinking in polyelectrolytes and an anisotropic distribution of oligoGs on interacting surfaces and provide a timescale for the formation and relaxation of alginate gels at the single crosslink level.

  5. Alternative interpretation for the active zones of Cuba

    Rodríguez, Mario Octavio Cotilla

    2014-11-01

    An alternative explanation to the seismoactivity of Cuban faults is presented. The model is a consequence of the interaction between Caribbean and North American plates. It is made with 12 geodynamic cells form by a set of 13 active faults and their 14 areas of intersection. These cells are recognized morpho-structural blocks. The area between Eastern Matanzas and Western Cauto-Nipe is excluded because of the low level of seismic information. Cuba has two types of seismogenetic structures: faults and intersection of faults.

  6. Postsynaptic actin regulates active zone spacing and glutamate receptor apposition at the Drosophila neuromuscular junction.

    Blunk, Aline D; Akbergenova, Yulia; Cho, Richard W; Lee, Jihye; Walldorf, Uwe; Xu, Ke; Zhong, Guisheng; Zhuang, Xiaowei; Littleton, J Troy

    2014-07-01

    Synaptic communication requires precise alignment of presynaptic active zones with postsynaptic receptors to enable rapid and efficient neurotransmitter release. How transsynaptic signaling between connected partners organizes this synaptic apparatus is poorly understood. To further define the mechanisms that mediate synapse assembly, we carried out a chemical mutagenesis screen in Drosophila to identify mutants defective in the alignment of active zones with postsynaptic glutamate receptor fields at the larval neuromuscular junction. From this screen we identified a mutation in Actin 57B that disrupted synaptic morphology and presynaptic active zone organization. Actin 57B, one of six actin genes in Drosophila, is expressed within the postsynaptic bodywall musculature. The isolated allele, act(E84K), harbors a point mutation in a highly conserved glutamate residue in subdomain 1 that binds members of the Calponin Homology protein family, including spectrin. Homozygous act(E84K) mutants show impaired alignment and spacing of presynaptic active zones, as well as defects in apposition of active zones to postsynaptic glutamate receptor fields. act(E84K) mutants have disrupted postsynaptic actin networks surrounding presynaptic boutons, with the formation of aberrant actin swirls previously observed following disruption of postsynaptic spectrin. Consistent with a disruption of the postsynaptic actin cytoskeleton, spectrin, adducin and the PSD-95 homolog Discs-Large are all mislocalized in act(E84K) mutants. Genetic interactions between act(E84K) and neurexin mutants suggest that the postsynaptic actin cytoskeleton may function together with the Neurexin-Neuroligin transsynaptic signaling complex to mediate normal synapse development and presynaptic active zone organization.

  7. CHEMICAL ACTIVATION OF MOLECULES BY METALS: EXPERIMENTAL STUDIES OF ELECTRON DISTRIBUTIONS AND BONDING

    LICHTENBERGER, DENNIS L.

    2002-03-26

    This research program is directed at obtaining detailed experimental information on the electronic interactions between metals and organic molecules. These interactions provide low energy pathways for many important chemical and catalytic processes. A major feature of the program is the continued development and application of our special high-resolution valence photoelectron spectroscopy (UPS), and high-precision X-ray core photoelectron spectroscopy (XPS) instrumentation for study of organometallic molecules in the gas phase. The study involves a systematic approach towards understanding the interactions and activation of bound carbonyls, C-H bonds, methylenes, vinylidenes, acetylides, alkenes, alkynes, carbenes, carbynes, alkylidenes, alkylidynes, and others with various monometal, dimetal, and cluster metal species. Supporting ligands include -aryls, alkoxides, oxides, and phosphines. We are expanding our studies of both early and late transition metal species and electron-rich and electron-poor environments in order to more completely understand the electronic factors that serve to stabilize particular organic fragments and intermediates on metals. Additional new directions for this program are being taken in ultra-high vacuum surface UPS, XPS, scanning tunneling microscopy (STM) and atomic force microscopy (AFM) experiments on both physisorbed and chemisorbed organometallic thin films. The combination of these methods provides additional electronic structure information on surface-molecule and molecule-molecule interactions. A very important general result emerging from this program is the identification of a close relationship between the ionization energies of the species and the thermodynamics of the chemical and catalytic reactions of these systems.

  8. Determination of absolute configuration of chiral molecules using vibrational optical activity: a review.

    He, Yanan; Wang, Bo; Dukor, Rina K; Nafie, Laurence A

    2011-07-01

    Determination of the absolute handedness, known as absolute configuration (AC), of chiral molecules is an important step in any field related to chirality, especially in the pharmaceutical industry. Vibrational optical activity (VOA) has become a powerful tool for the determination of the AC of chiral molecules in the solution state after nearly forty years of evolution. VOA offers a novel alternative, or supplement, to X-ray crystallography, permitting AC determinations on neat liquid, oil, and solution samples without the need to grow single crystals of the pure chiral sample molecules as required for X-ray analysis. By comparing the sign and intensity of the measured VOA spectrum with the corresponding ab initio density functional theory (DFT) calculated VOA spectrum of a chosen configuration, one can unambiguously assign the AC of a chiral molecule. Comparing measured VOA spectra with calculated VOA spectra of all the conformers can also provide solution-state conformational populations. VOA consists of infrared vibrational circular dichroism (VCD) and vibrational Raman optical activity (ROA). Currently, VCD is used routinely by researchers in a variety of backgrounds, including molecular chirality, asymmetric synthesis, chiral catalysis, drug screening, pharmacology, and natural products. Although the application of ROA in AC determination lags behind that of VCD, with the recent implementation of ROA subroutines in commercial quantum chemistry software, ROA will in the future complement VCD for AC determination. In this review, the basic principles of the application of VCD to the determination of absolute configuration in chiral molecules are described. The steps required for VCD spectral measurement and calculation are outlined, followed by brief descriptions of recently published papers reporting the determination of AC in small organic, pharmaceutical, and natural product molecules.

  9. Aqueous phase adsorption of different sized molecules on activated carbon fibers: Effect of textural properties.

    Prajapati, Yogendra N; Bhaduri, Bhaskar; Joshi, Harish C; Srivastava, Anurag; Verma, Nishith

    2016-07-01

    The effect that the textural properties of rayon-based activated carbon fibers (ACFs), such as the BET surface area and pore size distribution (PSD), have on the adsorption of differently sized molecules, namely, brilliant yellow (BY), methyl orange (MO) and phenol (PH), was investigated in the aqueous phase. ACF samples with different BET areas and PSDs were produced by steam-activating carbonized fibers for different activation times (0.25, 0.5, and 1 h). The samples activated for 0.25 h were predominantly microporous, whereas those activated for relatively longer times contained hierarchical micro-mesopores. The adsorption capacities of the ACFs for the adsorbate increased with increasing BET surface area and pore volume, and ranged from 51 to 1306 mg/g depending on the textural properties of the ACFs and adsorbate size. The adsorption capacities of the hierarchical ACF samples followed the order BY > MO > PH. Interestingly, the number of molecules adsorbed by the ACFs followed the reverse order: PH > MO > BY. This anomaly was attributed to the increasing molecular weight of the PH, MO and BY molecules. The equilibrium adsorption data were described using the Langmuir isotherm. This study shows that suitable textural modifications to ACFs are required for the efficient aqueous phase removal of an adsorbate.

  10. [Characteristics of soil organic carbon and enzyme activities in soil aggregates under different vegetation zones on the Loess Plateau].

    Li, Xin; Ma, Rui-ping; An, Shao-shan; Zeng, Quan-chao; Li, Ya-yun

    2015-08-01

    In order to explore the distribution characteristics of organic carbon of different forms and the active enzymes in soil aggregates with different particle sizes, soil samples were chosen from forest zone, forest-grass zone and grass zone in the Yanhe watershed of Loess Plateau to study the content of organic carbon, easily oxidized carbon, and humus carbon, and the activities of cellulase, β-D-glucosidase, sucrose, urease and peroxidase, as well as the relations between the soil aggregates carbon and its components with the active soil enzymes were also analyzed. It was showed that the content of organic carbon and its components were in order of forest zone > grass zone > forest-grass zone, and the contents of three forms of organic carbon were the highest in the diameter group of 0.25-2 mm. The content of organic carbon and its components, as well as the activities of soil enzymes were higher in the soil layer of 0-10 cm than those in the 10-20 cm soil layer of different vegetation zones. The activities of cellulase, β-D-glucosidase, sucrose and urease were in order of forest zone > grass zone > forest-grass zone. The peroxidase activity was in order of forest zone > forest-grass zone > grass zone. The activities of various soil enzymes increased with the decreasing soil particle diameter in the three vegetation zones. The activities of cellulose, peroxidase, sucrose and urease had significant positive correlations with the contents of various forms of organic carbon in the soil aggregates.

  11. Discovery of Small Molecules for Fluorescent Detection of Complement Activation Product C3d.

    Gorham, Ronald D; Nuñez, Vicente; Lin, Jung-Hsin; Rooijakkers, Suzan H M; Vullev, Valentine I; Morikis, Dimitrios

    2015-12-24

    Complement activation plays a major role in many acute and chronic inflammatory conditions. C3d, a terminal product of complement activation, remains covalently attached to cells and is an excellent biomarker of complement-mediated inflammation. We employed a virtual high-throughput screening protocol to identify molecules with predicted binding to complement C3d and with intrinsic fluorescence properties to enable detection. Pharmacophore models were developed based on known C3d-ligand interactions and information from computational analysis of structural and molecular dynamics data. Iterative pharmacophore-based virtual screening was performed to identify druglike molecules with physicochemical similarity to the natural C3d ligand CR2. Hits from the pharmacophore screens were docked to C3d and ranked based on predicted binding free energies. Top-ranked molecules were selected for experimental validation of binding affinity to C3d, using microscale thermophoresis, and for their suitability to become molecular imaging agents, using fluorescence spectroscopy. This work serves as a foundation for identifying additional fluorescent molecules with high-affinity for C3d that will subsequently be explored as noninvasive in vivo diagnostics of complement-mediated inflammation, for spatiotemporal monitoring of disease progression, and for targeting therapeutics to sites of inflammation.

  12. Matrine inhibits the expression of adhesion molecules in activated vascular smooth muscle cells.

    Liu, Jun; Zhang, Lihua; Ren, Yingang; Gao, Yanli; Kang, Li; Lu, Shaoping

    2016-03-01

    Atherosclerosis is a chronic inflammatory disease associated with increased expression of adhesion molecules in vascular smooth muscle cells (VSMCs). Matrine is a main active ingredient of Sophora flavescens roots, which are used to treat inflammatory diseases. However, the effects of matrine on the expression of adhesion molecules in VSMCs have largely remained elusive. Therefore, the present study investigated the effects of matrine on the expression of adhesion molecules in tumor necrosis factor (TNF)‑α‑stimulated human aortic smooth muscle cells (HASMCs). The results showed that matrine inhibited the expression of vascular cell adhesion molecule‑1 (VCAM‑1) and intercellular adhesion molecule‑1 (ICAM‑1) in TNF‑α‑stimulated HASMCs. Matrine markedly inhibited the TNF‑α‑induced expression of nuclear factor (NF)‑κB p65 and prevented the TNF‑α‑caused degradation of inhibitor of NF‑κB; it also inhibited TNF‑α‑induced activation of mitogen‑activated protein kinases (MAPKs). Furthermore, matrine inhibited the production of intracellular reactive oxygen species (ROS) in TNF‑α‑stimulated HASMCs. In conclusion, the results of the present study demonstrated that matrine inhibited the expression of VCAM‑1 and ICAM‑1 in TNF‑α‑stimulated HASMCs via the suppression of ROS production as well as NF‑κB and MAPK pathway activation. Therefore, matrine may have a potential therapeutic use for preventing the advancement of atherosclerotic lesions.

  13. Structure based discovery of small molecules to regulate the activity of human insulin degrading enzyme.

    Bilal Çakir

    Full Text Available BACKGROUND: Insulin-degrading enzyme (IDE is an allosteric Zn(+2 metalloprotease involved in the degradation of many peptides including amyloid-β, and insulin that play key roles in Alzheimer's disease (AD and type 2 diabetes mellitus (T2DM, respectively. Therefore, the use of therapeutic agents that regulate the activity of IDE would be a viable approach towards generating pharmaceutical treatments for these diseases. Crystal structure of IDE revealed that N-terminal has an exosite which is ∼30 Å away from the catalytic region and serves as a regulation site by orientation of the substrates of IDE to the catalytic site. It is possible to find small molecules that bind to the exosite of IDE and enhance its proteolytic activity towards different substrates. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we applied structure based drug design method combined with experimental methods to discover four novel molecules that enhance the activity of human IDE. The novel compounds, designated as D3, D4, D6, and D10 enhanced IDE mediated proteolysis of substrate V, insulin and amyloid-β, while enhanced degradation profiles were obtained towards substrate V and insulin in the presence of D10 only. CONCLUSION/SIGNIFICANCE: This paper describes the first examples of a computer-aided discovery of IDE regulators, showing that in vitro and in vivo activation of this important enzyme with small molecules is possible.

  14. Tsunamigenic potential of Mediterranean fault systems and active subduction zones

    Petricca, Patrizio; Babeyko, Andrey

    2016-04-01

    Since the North East Atlantic and Mediterranean Tsunami Warning System (NEAMTWS) is under development by the European scientific community, it becomes necessary to define guidelines for the characterization of the numerous parameters must be taken into account in a fair assessment of the risk. Definition of possible tectonic sources and evaluation of their potential is one of the principal issues. In this study we systematically evaluate tsunamigenic potential of up-to-now known real fault systems and active subduction interfaces in the NEAMTWS region. The task is accomplished by means of numerical modeling of tsunami generation and propagation. We have simulated all possible uniform-slip ruptures populating fault and subduction interfaces with magnitudes ranging from 6.5 up to expected Mmax. A total of 15810 individual ruptures were processed. For each rupture, a tsunami propagation scenario was computed in linear shallow-water approximation on 1-arc minute bathymetric grid (Gebco_08) implying normal reflection boundary conditions. Maximum wave heights at coastal positions (totally - 23236 points of interest) were recorded for four hours of simulation and then classified according to currently adopted warning level thresholds. The resulting dataset allowed us to classify the sources in terms of their tsunamigenic potential as well as to estimate their minimum tsunamigenic magnitude. Our analysis shows that almost every source in the Mediterranean Sea is capable to produce local tsunami at the advisory level (i.e., wave height > 20 cm) starting from magnitude values of Mw=6.6. In respect to the watch level (wave height > 50 cm), the picture is less homogeneous: crustal sources in south-west Mediterranean as well as East-Hellenic arc need larger magnitudes (around Mw=7.0) to trigger watch levels even at the nearby coasts. In the context of the regional warning (i.e., source-to-coast distance > 100 km) faults also behave more heterogeneously in respect to the minimum

  15. A New Perspective on the Radio Active Zone at The Galactic Center - Feedback from Nuclear Activities

    Zhao, Jun-Hui; Goss, W M

    2013-01-01

    Based on our deep image of Sgr A using broadband data observed with the Jansky VLA at 6 cm, we present a new perspective of the radio bright zone at the Galactic center. We further show the radio detection of the X-ray Cannonball, a candidate neutron star associated with the Galactic center SNR Sgr A East. The radio image is compared with the Chandra X-ray image to show the detailed structure of the radio counterparts of the bipolar X-ray lobes. The bipolar lobes are likely produced by the winds from the activities within Sgr A West, which could be collimated by the inertia of gas in the CND, or by the momentum driving of Sgr A*; and the poloidal magnetic fields likely play an important role in the collimation. The less-collimated SE lobe, in comparison to the NW one, is perhaps due to the fact that the Sgr A East SN might have locally reconfigured the magnetic field toward negative galactic latitudes. In agreement with the X-ray observations, the time-scale of ~ $1\\times10^4$ yr estimated for the outermost r...

  16. Application of terahertz spectroscopy for characterization of biologically active organic molecules in natural environment

    Karaliūnas, Mindaugas; Jakštas, Vytautas; Nasser, Kinan E.; Venckevičius, Rimvydas; Urbanowicz, Andrzej; Kašalynas, Irmantas; Valušis, Gintaras

    2016-09-01

    In this work, a comparative research of biologically active organic molecules in its natural environment using the terahertz (THz) time domain spectroscopy (TDS) and Fourier transform spectroscopy (FTS) systems is carried out. Absorption coefficient and refractive index of Nicotiana tabacum L. leaves containing nicotine, Cannabis sativa L. leaves containing tetrahydrocannabinol, and Humulu lupulus L. leaves containing α-acids, active organic molecules that obtain in natural environment, were measured in broad frequency range from 0.1 to 13 THz at room temperature. In the spectra of absorption coefficient the features were found to be unique for N. tabacum, C. sativa and H. lupulus. Moreover, those features can be exploited for identification of C. sativa sex and N. tabacum origin. The refractive index can be also used to characterize different species.

  17. Water molecule network and active site flexibility of apo protein tyrosine phosphatase 1B

    Pedersen, A.K.; Peters, Günther H.J.; Møller, K.B.;

    2004-01-01

    Protein tyrosine phosphatase 1B (PTP1B) plays a key role as a negative regulator of insulin and leptin signalling and is therefore considered to be an important molecular target for the treatment of type 2 diabetes and obesity. Detailed structural information about the structure of PTP1B, including...... the conformation and flexibility of active-site residues as well as the water-molecule network, is a key issue in understanding ligand binding and enzyme kinetics and in structure-based drug design. A 1.95 Angstrom apo PTP1B structure has been obtained, showing four highly coordinated water molecules in the active...... of PTP1B and form a novel basis for structure-based inhibitor design....

  18. Nanoscale charge transport in cytochrome c3/DNA network: Comparative studies between redox-active molecules

    Yamaguchi, Harumasa; Che, Dock-Chil; Hirano, Yoshiaki; Suzuki, Masayuki; Higuchi, Yoshiki; Matsumoto, Takuya

    2015-09-01

    The redox-active molecule of a cytochrome c3/DNA network exhibits nonlinear current-voltage (I-V) characteristics with a threshold bias voltage at low temperature and zero-bias conductance at room temperature. I-V curves for the cytochrome c3/DNA network are well matched with the Coulomb blockade network model. Comparative studies of the Mn12 cluster, cytochrome c, and cytochrome c3, which have a wide variety of redox potentials, indicate no difference in charge transport, which suggests that the conduction mechanism is not directly related to the redox states. The charge transport mechanism has been discussed in terms of the newly-formed electronic energy states near the Fermi level, induced by the ionic interaction between redox-active molecules with the DNA network.

  19. Anticancer molecule AS1411 exhibits low nanomolar antiviral activity against HIV-1.

    Métifiot, Mathieu; Amrane, Samir; Mergny, Jean-Louis; Andreola, Marie-Line

    2015-11-01

    During clinical trials, a number of fully characterized molecules are dropped along the way because they do not provide enough benefit for the patient. Some of them show limited side effects and might be of great use for other applications. AS1411 is a nucleolin-targeting aptamer that underwent phase II clinical trials as anticancer agent. Here, we show that AS1411 exhibits extremely potent antiviral activity and is therefore an attractive new lead as anti-HIV agent.

  20. Structure-property relationship of quinuclidinium surfactants--Towards multifunctional biologically active molecules.

    Skočibušić, Mirjana; Odžak, Renata; Štefanić, Zoran; Križić, Ivana; Krišto, Lucija; Jović, Ozren; Hrenar, Tomica; Primožič, Ines; Jurašin, Darija

    2016-04-01

    Motivated by diverse biological and pharmacological activity of quinuclidine and oxime compounds we have synthesized and characterized novel class of surfactants, 3-hydroxyimino quinuclidinium bromides with different alkyl chains lengths (CnQNOH; n=12, 14 and 16). The incorporation of non conventional hydroxyimino quinuclidinium headgroup and variation in alkyl chain length affects hydrophilic-hydrophobic balance of surfactant molecule and thereby physicochemical properties important for its application. Therefore, newly synthesized surfactants were characterized by the combination of different experimental techniques: X-ray analysis, potentiometry, electrical conductivity, surface tension and dynamic light scattering measurements, as well as antimicrobial susceptibility tests. Comprehensive investigation of CnQNOH surfactants enabled insight into structure-property relationship i.e., way in which the arrangement of surfactant molecules in the crystal phase correlates with their solution behavior and biologically activity. The synthesized CnQNOH surfactants exhibited high adsorption efficiency and relatively low critical micelle concentrations. In addition, all investigated compounds showed very potent and promising activity against Gram-positive and clinically relevant Gram-negative bacterial strains compared to conventional antimicrobial agents: tetracycline and gentamicin. The overall results indicate that bicyclic headgroup with oxime moiety, which affects both hydrophilicity and hydrophobicity of CnQNOH molecule in addition to enabling hydrogen bonding, has dominant effect on crystal packing and physicochemical properties. The unique structural features of cationic surfactants with hydroxyimino quinuclidine headgroup along with diverse biological activity have made them promising structures in novel drug discovery. Obtained fundamental understanding how combination of different functionalities in a single surfactant molecule affects its physicochemical

  1. Identification of intrinsic catalytic activity for electrochemical reduction of water molecules to generate hydrogen

    Shinagawa, Tatsuya

    2015-01-01

    Insufficient hydronium ion activities at near-neutral pH and under unbuffered conditions induce diffusion-limited currents for hydrogen evolution, followed by a reaction with water molecules to generate hydrogen at elevated potentials. The observed constant current behaviors at near neutral pH reflect the intrinsic electrocatalytic reactivity of the metal electrodes for water reduction. This journal is © the Owner Societies.

  2. Microbial respiration and extracellular enzyme activity in sediments from the Gulf of Mexico hypoxic zone

    This study explores the relationship between sediment chemistry (TC, TN, TP) and microbial respiration (DHA) and extracellular enzyme activity (EEA) across the Gulf of Mexico (GOM) hypoxic zone. TC, TN, and TP were all positively correlated with each other (r=0.19-0.68). DHA was ...

  3. The Legality of Foreign Military Activities in the Exclusive Economic Zone under UNCLOS

    Geng, Jing

    2012-01-01

    During negotiations for the United Nations Convention on the Law of the Sea (UNCLOS), military activities in another state's Exclusive Economic Zone (EEZ) were a point of contention. Currently, the issue remains controversial in state practice. UNCLOS attempts to balance the differing interests of c

  4. Small molecule inhibitors block Gas6-inducible TAM activation and tumorigenicity

    Kimani, Stanley G.; Kumar, Sushil; Bansal, Nitu; Singh, Kamalendra; Kholodovych, Vladyslav; Comollo, Thomas; Peng, Youyi; Kotenko, Sergei V.; Sarafianos, Stefan G.; Bertino, Joseph R.; Welsh, William J.; Birge, Raymond B.

    2017-01-01

    TAM receptors (Tyro-3, Axl, and Mertk) are a family of three homologous type I receptor tyrosine kinases that are implicated in several human malignancies. Overexpression of TAMs and their major ligand Growth arrest-specific factor 6 (Gas6) is associated with more aggressive staging of cancers, poorer predicted patient survival, acquired drug resistance and metastasis. Here we describe small molecule inhibitors (RU-301 and RU-302) that target the extracellular domain of Axl at the interface of the Ig-1 ectodomain of Axl and the Lg-1 of Gas6. These inhibitors effectively block Gas6-inducible Axl receptor activation with low micromolar IC50s in cell-based reporter assays, inhibit Gas6-inducible motility in Axl-expressing cell lines, and suppress H1299 lung cancer tumor growth in a mouse xenograft NOD-SCIDγ model. Furthermore, using homology models and biochemical verifications, we show that RU301 and 302 also inhibit Gas6 inducible activation of Mertk and Tyro3 suggesting they can act as pan-TAM inhibitors that block the interface between the TAM Ig1 ectodomain and the Gas6 Lg domain. Together, these observations establish that small molecules that bind to the interface between TAM Ig1 domain and Gas6 Lg1 domain can inhibit TAM activation, and support the further development of small molecule Gas6-TAM interaction inhibitors as a novel class of cancer therapeutics. PMID:28272423

  5. Early-Late Heterobimetallic Complexes Linked by Phosphinoamide Ligands. Tuning Redox Potentials and Small Molecule Activation

    Thomas, Christine M. [Brandeis Univ., Waltham, MA (United States)

    2015-08-01

    Recent attention in the chemical community has been focused on the energy efficient and environmentally benign conversion of abundant small molecules (CO2, H2O, etc.) to useful liquid fuels. This project addresses these goals by examining fundamental aspects of catalyst design to ultimately access small molecule activation processes under mild conditions. Specifically, Thomas and coworkers have targetted heterobimetallic complexes that feature metal centers with vastly different electronic properties, dictated both by their respective positions on the periodic table and their coordination environment. Unlike homobimetallic complexes featuring identical or similar metals, the bonds between metals in early/late heterobimetallics are more polarized, with the more electron-rich late metal center donating electron density to the more electron-deficient early metal center. While metal-metal bonds pose an interesting strategy for storing redox equivalents and stabilizing reactive metal fragments, the polar character of metal-metal bonds in heterobimetallic complexes renders these molecules ideally poised to react with small molecule substrates via cleavage of energy-rich single and double bonds. In addition, metal-metal interactions have been shown to dramatically affect redox potentials and promote multielectron redox activity, suggesting that metal-metal interactions may provide a mechanism to tune redox potentials and access substrate reduction/activation at mild overpotentials. This research project has provided a better fundamental understanding of how interactions between transition metals can be used as a strategy to promote and/or control chemical transformations related to the clean production of fuels. While this project focused on the study of homogeneous systems, it is anticipated that the broad conclusions drawn from these investigations will be applicable to heterogeneous catalysis as well, particularly on heterogeneous processes that occur at interfaces in

  6. Green Oxygenation Degradation of Rhodamine B by Using Activated Molecule Oxygen

    Ke Jian DENG; Fei HUANG; Duo Yuan WANG; Zheng He PENG; Yun Hong ZHOU

    2004-01-01

    Iron(Ⅱ) tetra-(1,4-dithin)-porphyrazine, (FePz(dtn)4) is able to activate molecule oxygen for oxygenation degradation of rhodamine B (RhB) in an extensive pH region without light excitation. Experiments indicate that the RhB can be degraded nearly 52% in alkaline aqueous solution, bubbling with dioxygen for seven hours in the presence of FePz(dtn)4 and the hydrogen peroxides as an actve intermediate were determined by DPD method. The catalyst is recyclable and the catalyst activity was maintained after l0 recycles.

  7. Single-Molecule Imaging Reveals the Activation Dynamics of Intracellular Protein Smad3 on Cell Membrane

    Li, Nan; Yang, Yong; He, Kangmin; Zhang, Fayun; Zhao, Libo; Zhou, Wei; Yuan, Jinghe; Liang, Wei; Fang, Xiaohong

    2016-09-01

    Smad3 is an intracellular protein that plays a key role in propagating transforming growth factor β (TGF-β) signals from cell membrane to nucleus. However whether the transient process of Smad3 activation occurs on cell membrane and how it is regulated remains elusive. Using advanced live-cell single-molecule fluorescence microscopy to image and track fluorescent protein-labeled Smad3, we observed and quantified, for the first time, the dynamics of individual Smad3 molecules docking to and activation on the cell membrane. It was found that Smad3 docked to cell membrane in both unstimulated and stimulated cells, but with different diffusion rates and dissociation kinetics. The change in its membrane docking dynamics can be used to study the activation of Smad3. Our results reveal that Smad3 binds with type I TGF-β receptor (TRI) even in unstimulated cells. Its activation is regulated by TRI phosphorylation but independent of receptor endocytosis. This study offers new information on TGF-β/Smad signaling, as well as a new approach to investigate the activation of intracellular signaling proteins for a better understanding of their functions in signal transduction.

  8. Ethosomes for the delivery of anti-HSV-1 molecules: preparation, characterization and in vitro activity.

    Cortesi, R; Ravani, L; Zaid, A N; Menegatti, E; Romagnoli, R; Drechsler, M; Esposito, E

    2010-10-01

    This paper describes the production, characterization and in vitro activity of ethosomes containing two molecules with antiviral activity, such as acyclovir (ACY) and N1-beta-D-ribofuranosyl-pyrazole [3,4d]pyridazin-7(6p-chlorine-phenyl)-one nucleoside (N1CP). Ethosomes were prepared and morphologically characterized by Cryo-TEM. The encapsulation efficiency was 92.3 +/- 2.5% for ACY and 94.2 +/- 2.8% for N1CP. The release of the drug from vesicles, determined by a Franz cell method, indicated that both drugs were released in a controlled manner. In order to possibly guarantee the stability during long-term storage ethosome suspensions was freeze-dried. It was found that the freeze-dried ethosomes' cakes were compact, glassy characterized by low density and quick re-hydration. However, the storage time slightly influences the percentage of drug encapsulation within ethosomes showing a drug leakage after re-hydration around 10%. The antiviral activity against HSV-1 of both drugs was tested by plaque reduction assay in monolayer cultures of Vero cells. Data showed that ethosomes allowed a reduction of the ED50 of N1CP evidencing an increase of its antiviral activity. However, ACY remains more active than N1CP. No differences are appreciable between drug-containing ethosomes before and after freeze-drying. Taken together these results, ethosomal formulation could be possibly proposed as mean for topical administration of anti-herpetic molecules.

  9. Small molecules activating TrkB receptor for treating a variety of CNS disorders.

    Zeng, Yan; Wang, Xiaonan; Wang, Qiang; Liu, Shumin; Hu, Xiamin; McClintock, Shawn M

    2013-11-01

    The brain-derived neurotrophic factor (BDNF) and its high affinity receptor tropomyosin-receptor-kinase B (TrkB) play a critical role in neuronal differentiation and survival, synapse plasticity, and memory. Indeed, both have been implicated in the pathophysiology of numerous diseases. Although the remarkable therapeutic potential of BDNF has generated much research over the past decade, the poor pharmacokinetics and adverse side effect profile have limited its clinical usefulness of BDNF. Small compounds that mimic BDNF's neurotrophic signaling and overcome the pharmacokinetic and side effect barriers may have greater therapeutic potential. The purpose of this review is to provide a survey of the various strategies taken towards the development of small molecule mimetics for BDNF and the selective TrkB agonist. A particular focus was placed on TrkB agonist 7, 8-dihydroxyflavone, which modulates multiple functions and has demonstrated remarkable therapeutic efficacy in a variety of central nervous system disease models. Two other small molecules included in this review are adenosine A2A receptor agonists that indirectly activate TrkB, and TrkB binding domains of BDNF, loop II-LM22A compounds that directly activate TrkB. These alternative molecules have shown promise in preclinical studies and may be included in prospective clinical investigations.

  10. Theoretical investigation of the degradation mechanisms in host and guest molecules used in OLED active layers

    Winget, Paul

    2014-10-08

    A feature of OLEDs that has to date received little attention is the prediction of the stability of the molecules involved in the electrical and optical processes. Here, we present computational results intended to aid in the development of stable systems. We identify degradation pathways and define new strategies to guide the synthesis of stable materials for OLED applications for both phosphorescent emitters and organic host materials. The chemical reactivity of these molecules in the active layers of the devices is further complicated by the fact that, during operation, they can be either oxidized or reduced (as they localize a hole or an electron) in addition to forming both singlet and triplet excitons. © (2014) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.

  11. Bruchpilot and Synaptotagmin collaborate to drive rapid glutamate release and active zone differentiation

    Mila M Paul

    2015-02-01

    Full Text Available The active zone (AZ protein Bruchpilot (Brp is essential for rapid glutamate release at Drosophila melanogaster neuromuscular junctions (NMJs. Quantal time course and measurements of action potential-waveform suggest that presynaptic fusion mechanisms are altered in brp null mutants (brp69. This could account for their increased evoked excitatory postsynaptic current (EPSC delay and rise time (by about one millisecond. To test the mechanism of release protraction at brp69 AZs, we performed knock-down of Synaptotagmin-1 (Syt via RNAi (sytKD in wildtype (wt, brp69 and rab3 null mutants (rab3rup, where Brp is concentrated at a small number of AZs. At wt and rab3rup synapses, sytKD lowered EPSC amplitude while increasing rise time and delay, consistent with the role of Syt as a release sensor. In contrast, sytKD did not alter EPSC amplitude at brp69 synapses, but shortened delay and rise time. In fact, following sytKD, these kinetic properties were strikingly similar in wt and brp69, which supports the notion that Syt protracts release at brp69 synapses. To gain insight into this surprising role of Syt at brp69 AZs, we analyzed the structural and functional differentiation of synaptic boutons at the NMJ. At ’tonic’ type Ib motor neurons, distal boutons contain more AZs, more Brp proteins per AZ and show elevated and accelerated glutamate release compared to proximal boutons. The functional differentiation between proximal and distal boutons is Brp-dependent and reduced after sytKD. Notably, sytKD boutons are smaller, contain fewer Brp positive AZs and these are of similar number in proximal and distal boutons. In addition, super-resolution imaging via dSTORM revealed that sytKD increases the number and alters the spatial distribution of Brp molecules at AZs, while the gradient of Brp proteins per AZ is diminished. In summary, these data demonstrate that normal structural and functional differentiation of Drosophila AZs requires concerted action

  12. Stellar Activity Masquerading as Planets in the Habitable Zone of the M dwarf Gliese 581

    Robertson, Paul; Endl, Michael; Roy, Arpita

    2014-01-01

    The M dwarf Gliese 581 is believed to host four planets, including one (GJ 581d) near the habitable zone that could possibly support liquid water on its surface if it is a rocky planet. The detection of another habitable-zone planet--GJ 581g--is disputed, as its significance depends on the eccentricity assumed for d. Analyzing stellar activity using the H-alpha line, we measure a stellar rotation period of 130+/-2 days and a correlation for H-alpha modulation with radial velocity. Correcting for activity greatly diminishes the signal of GJ 581d (to 1.5 sigma), while significantly boosting the signals of the other known super-Earth planets. GJ 581d does not exist, but is an artifact of stellar activity which, when incompletely corrected, causes the false detection of planet g.

  13. Functional inactivation of a fraction of excitatory synapses in mice deficient for the active zone protein bassoon

    Altrock, Wilko D; tom Dieck, Susanne; Sokolov, Maxim;

    2003-01-01

    Mutant mice lacking the central region of the presynaptic active zone protein Bassoon were generated to establish the role of this protein in the assembly and function of active zones as sites of synaptic vesicle docking and fusion. Our data show that the loss of Bassoon causes a reduction in nor...

  14. Acoustic input and efferent activity regulate the expression of molecules involved in cochlear micromechanics

    Verónica eLamas

    2015-01-01

    Full Text Available Electromotile activity in auditory outer hair cells (OHCs is essential for sound amplification. It relies on the highly specialized membrane motor protein prestin, and its interactions with the cytoskeleton. It is believed that the expression of prestin and related molecules involved in OHC electromotility may be dynamically regulated by signals from the acoustic environment. However little is known about the nature of such signals and how they affect the expression of molecules involved in electromotility in OHCs. We show evidence that prestin oligomerization is regulated, both at short and relatively long term, by acoustic input and descending efferent activity originating in the cortex, likely acting in concert. Unilateral removal of the middle ear ossicular chain reduces levels of trimeric prestin, particularly in the cochlea from the side of the lesion, whereas monomeric and dimeric forms are maintained or even increased in particular in the contralateral side, as shown in Western blots. Unilateral removal of the auditory cortex (AC, which likely causes an imbalance in descending efferent activity on the cochlea, also reduces levels of trimeric and tetrameric forms of prestin in the side ipsilateral to the lesion, whereas in the contralateral side prestin remains unaffected, or even increased in the case of trimeric and tetrameric forms. As far as efferent inputs are concerned, unilateral ablation of the AC up-regulates the expression of α10 nAChR transcripts in the cochlea. Limited, unbalanced efferent activity after unilateral AC removal, also affects prestin and β-actin mRNA levels. These findings support that the concerted action of acoustic and efferent inputs to the cochlea is needed to regulate the expression of major molecules involved in OHC electromotility, both at the transcriptional and posttranscriptional levels.

  15. Small-molecule activation of SERCA2a SUMOylation for the treatment of heart failure

    Kho, Changwon; Lee, Ahyoung; Jeong, Dongtak; Oh, Jae Gyun; Gorski, Przemek A.; Fish, Kenneth; Sanchez, Roberto; DeVita, Robert J.; Christensen, Geir; Dahl, Russell; Hajjar, Roger J.

    2015-01-01

    Decreased activity and expression of the cardiac sarcoplasmic reticulum calcium ATPase (SERCA2a), a critical pump regulating calcium cycling in cardiomyocyte, are hallmarks of heart failure. We have previously described a role for the small ubiquitin-like modifier type 1 (SUMO-1) as a regulator of SERCA2a and have shown that gene transfer of SUMO-1 in rodents and large animal models of heart failure restores cardiac function. Here, we identify and characterize a small molecule, N106, which increases SUMOylation of SERCA2a. This compound directly activates the SUMO-activating enzyme, E1 ligase, and triggers intrinsic SUMOylation of SERCA2a. We identify a pocket on SUMO E1 likely to be responsible for N106's effect. N106 treatment increases contractile properties of cultured rat cardiomyocytes and significantly improves ventricular function in mice with heart failure. This first-in-class small-molecule activator targeting SERCA2a SUMOylation may serve as a potential therapeutic strategy for treatment of heart failure. PMID:26068603

  16. Formation of neutral disk-like zone around the active hot stars in symbiotic binaries

    Carikova, Zuzana

    2011-01-01

    In this contribution we present the ionization structure in the enhanced wind from the hot star in symbiotic binaries during active phases. Rotation of the hot star leads to the compression of the outflowing material towards its equatorial plane. As a result a neutral disk-like zone around the active hot star near the orbital plane is created. We modelled the compression of the wind using the wind compression model. Further, we calculated the neutral disk-like zone in the enhanced wind from the hot star using the equation of the photoionization equilibrium. The presence of such neutral disk-like zones was also suggested on the basis of the modelling the spectral energy distribution of symbiotic binaries. We confront the calculated ionization structures in the enhanced wind from the hot star with the observations. We found that the calculated column density of the neutral hydrogen atoms in the neutral disk-like zone and the emission measure of the ionized part of the wind from the hot star are in a good agreem...

  17. Influence of the water molecules near surface of viral protein on virus activation process

    O, Shepelenko S; S, Salnikov A; V, Rak S; P, Goncharova E; B, Ryzhikov A, E-mail: shep@vector.nsc.r, E-mail: shep@ngs.r [Federal State Research Institution State Research Center of Virology and Biotechnology VECTOR of the Federal Service for Surveillance in Consumer Rights Protection and Human Well-being (FSRI SRC VB VECTOR) Koltsovo, Novosibirsk Region (Russian Federation)

    2009-06-01

    The infection of a cell with influenza virus comprises the stages of receptor binding to the cell membrane, endocytosis of virus particle, and fusion of the virus envelope and cell endosome membrane, which is determined by the conformational changes in hemagglutinin, a virus envelope protein, caused by pH decrease within the endosome. The pH value that induces conformation rearrangements of hemagglutinin molecule considerably varies for different influenza virus strains, first and foremost, due to the differences in amino acid structure of the corresponding proteins. The main goal of this study was to construct a model making it possible to assess the critical pH value characterizing the fusogenic activity of influenza virus hemagglutinin from the data on hemagglutinin structure and experimental verification of this model. Under this model, we assume that when the electrostatic force between interacting hemagglutinin molecules in the virus envelop exceeds a certain value, the hemagglutinin HA1 subunits are arranged so that they form a cavity sufficient for penetration of water molecules. This event leads to an irreversible hydration of the inner fragments of hemagglutinin molecule in a trimer and to the completion of conformational changes. The geometry of electrostatic field in hemagglutinin trimer was calculated taking into account the polarization effects near the interface of two dielectrics, aqueous medium and protein macromolecule. The critical pH values for the conformational changes in hemagglutinin were measured by the erythrocyte hemolysis induced by influenza virus particles when decreasing pH. The critical pH value conditionally separating the pH range into the regions with and without the conformational changes was calculated for several influenza virus H1N1 and H3N2 strains based on the data on the amino acid structure of the corresponding hemagglutinin molecules. Comparison of the theoretical and experimental values of critical pH values for

  18. Force-activated reactivity switch in a bimolecular chemical reaction at the single molecule level

    Szoszkiewicz, Robert; Garcia-Manyes, Sergi; Liang, Jian; Kuo, Tzu-Ling; Fernandez, Julio M.

    2010-03-01

    Mechanical force can deform the reacting molecules along a well-defined direction of the reaction coordinate. However, the effect of mechanical force on the free-energy surface that governs a chemical reaction is still largely unknown. The combination of protein engineering with single-molecule AFM force-clamp spectroscopy allows us to study the influence of mechanical force on the rate at which a protein disulfide bond is reduced by some reducing agents in a bimolecular substitution reaction (so-called SN2). We found that cleavage of a protein disulfide bond by hydroxide anions exhibits an abrupt reactivity ``switch'' at 500 pN, after which the accelerating effect of force on the rate of an SN2 chemical reaction greatly diminishes. We propose that an abrupt force-induced conformational change of the protein disulfide bond shifts its ground state, drastically changing its reactivity in SN2 chemical reactions. Our experiments directly demonstrate the action of a force-activated switch in the chemical reactivity of a single molecule. References: Sergi Garcia-Manyes, Jian Liang, Robert Szoszkiewicz, Tzu-Ling Kuo and Julio M. Fernandez, Nature Chemistry, 1, 236-242, 2009.

  19. High Throughput Extraction of Plant, Marine and Fungal Specimens for Preservation of Biologically Active Molecules

    Thomas G. McCloud

    2010-06-01

    Full Text Available The Developmental Therapeutics Program (DTP of the U.S. National Cancer Institute (NCI, at its NCI-Frederick facility, has built perhaps the largest and most diverse natural products screening library in the world for drug discovery. Composed of plant, marine organism and microbial extracts, it currently contains in excess of 230,000 unique materials. From the inception of this program to identify new anticancer chemotherapeutics from natural products sources in 1987, two extracts have been sequentially prepared from each specimen: one produced by organic solvent extraction, which yields a complex material that contains non- to moderately polar small molecules, and a water-soluble extract, a milieu largely unexplored for useful drugs in earlier years, which contains polar small to medium-sized molecules. Plant specimens and microbial ferments are extracted by modified traditional methods, while the method developed to produce extracts from marine organisms is unique and very different from that used by marine natural products chemists previously, but again yields both an organic solvent soluble and a water soluble material for inclusion into the screening library. Details of high throughput extract production for preservation of biologically active molecules are presented.

  20. 78 FR 16465 - Foreign-Trade Zone 7-Mayaguez, Puerto Rico, Authorization of Production Activity, Pepsi Cola...

    2013-03-15

    ... Activity, Pepsi Cola Puerto Rico Distributing, LLC (Soft Drink and Fruit Drink Beverages), Toa Baja, Puerto... notification of proposed production activity to the Foreign-Trade Zones (FTZ) Board on behalf of Pepsi...

  1. In vitro and in vivo activity of a novel antifungal small molecule against Candida infections.

    Sarah Sze Wah Wong

    Full Text Available Candida is the most common fungal pathogen of humans worldwide and has become a major clinical problem because of the growing number of immunocompromised patients, who are susceptible to infection. Moreover, the number of available antifungals is limited, and antifungal-resistant Candida strains are emerging. New and effective antifungals are therefore urgently needed. Here, we discovered a small molecule with activity against Candida spp. both in vitro and in vivo. We screened a library of 50,240 small molecules for inhibitors of yeast-to-hypha transition, a major virulence attribute of Candida albicans. This screening identified 20 active compounds. Further examination of the in vitro antifungal and anti-biofilm properties of these compounds, using a range of Candida spp., led to the discovery of SM21, a highly potent antifungal molecule (minimum inhibitory concentration (MIC 0.2-1.6 µg/ml. In vitro, SM21 was toxic to fungi but not to various human cell lines or bacterial species and was active against Candida isolates that are resistant to existing antifungal agents. Moreover, SM21 was relatively more effective against biofilms of Candida spp. than the current antifungal agents. In vivo, SM21 prevented the death of mice in a systemic candidiasis model and was also more effective than the common antifungal nystatin at reducing the extent of tongue lesions in a mouse model of oral candidiasis. Propidium iodide uptake assay showed that SM21 affected the integrity of the cell membrane. Taken together, our results indicate that SM21 has the potential to be developed as a novel antifungal agent for clinical use.

  2. Small-molecule activators of insulin-degrading enzyme discovered through high-throughput compound screening.

    Christelle Cabrol

    Full Text Available BACKGROUND: Hypocatabolism of the amyloid beta-protein (Abeta by insulin-degrading enzyme (IDE is implicated in the pathogenesis of Alzheimer disease (AD, making pharmacological activation of IDE an attractive therapeutic strategy. However, it has not been established whether the proteolytic activity of IDE can be enhanced by drug-like compounds. METHODOLOGY/PRINCIPAL FINDINGS: Based on the finding that ATP and other nucleotide polyphosphates modulate IDE activity at physiological concentrations, we conducted parallel high-throughput screening campaigns in the absence or presence of ATP and identified two compounds--designated Ia1 and Ia2--that significantly stimulate IDE proteolytic activity. Both compounds were found to interfere with the crosslinking of a photoaffinity ATP analogue to IDE, suggesting that they interact with a bona fide ATP-binding domain within IDE. Unexpectedly, we observed highly synergistic activation effects when the activity of Ia1 or Ia2 was tested in the presence of ATP, a finding that has implications for the mechanisms underlying ATP-mediated activation of IDE. Notably, Ia1 and Ia2 activated the degradation of Abeta by approximately 700% and approximately 400%, respectively, albeit only when Abeta was presented in a mixture also containing shorter substrates. CONCLUSIONS/SIGNIFICANCE: This study describes the first examples of synthetic small-molecule activators of IDE, showing that pharmacological activation of this important protease with drug-like compounds is achievable. These novel activators help to establish the putative ATP-binding domain as a key modulator of IDE proteolytic activity and offer new insights into the modulatory action of ATP. Several larger lessons abstracted from this screen will help inform the design of future screening campaigns and facilitate the eventual development of IDE activators with therapeutic utility.

  3. Complement activation and kidney injury molecule-1-associated proximal tubule injury in severe preeclampsia.

    Burwick, Richard M; Easter, Sarah Rae; Dawood, Hassan Y; Yamamoto, Hidemi S; Fichorova, Raina N; Feinberg, Bruce B

    2014-10-01

    Kidney injury with proteinuria is a characteristic feature of preeclampsia, yet the nature of injury in specific regions of the nephron is incompletely understood. Our study aimed to use existing urinary biomarkers to describe the pattern of kidney injury and proteinuria in pregnancies affected by severe preeclampsia. We performed a case-control study of pregnant women from Brigham and Women's Hospital from 2012 to 2013. We matched cases of severe preeclampsia (n=25) 1:1 by parity and gestational age to 2 control groups with and without chronic hypertension. Urinary levels of kidney injury molecule-1 and complement components (C3a, C5a, and C5b-9) were measured by enzyme-linked immunosorbent assay, and other markers (albumin, β2 microglobulin, cystatin C, epithelial growth factor, neutrophil gelatinase-associated lipocalin, osteopontin, and uromodulin) were measured simultaneously with a multiplex electrochemiluminescence assay. Median values between groups were compared with the Wilcoxon signed-rank test and correlations with Spearman correlation coefficient. Analysis of urinary markers revealed higher excretion of albumin and kidney injury molecule-1 and lower excretion of neutrophil gelatinase-associated lipocalin and epithelial growth factor in severe preeclampsia compared with chronic hypertension and healthy controls. Among subjects with severe preeclampsia, urinary excretion of complement activation products correlated most closely with kidney injury molecule-1, a specific marker of proximal tubule injury (C5a: r=0.60; P=0.001; and C5b-9: r=0.75; Pkidney injury in severe preeclampsia that is characterized by glomerular impairment and complement-mediated inflammation and injury, possibly localized to the proximal tubule in association with kidney injury molecule-1.

  4. Identification of a small molecule activator of novel PKCs for promoting glucose-dependent insulin secretion

    Shuai Han; Heling Pan; Jianhua Zhang; Li Tan; Dawei Ma; Junying Yuan; Jia-Rui Wu

    2011-01-01

    Using an image-based screen for small molecules that can affect Golgi morphology, we identify a small molecule,Sioc145, which can enlarge the Golgi compartments and promote protein secretion. More importantly, Siocl45 potentiates insulin secretion in a glucose-dependent manner. We show that Sioc145 selectively activates novel protein kinase Cs (nPKCs; δ and ε) but not conventional PKCs;cPKCs; a, βI and βll) in INS-1E insulinoma cells. In contrast, PMA, a non-selective activator of cPKCs and nPKCs, promotes insulin secretion independent of glucose concentrations. Furthermore, we demonstrate that Sioc145 and PMA show differential abilities in depolarizing the cell membrane, and suggest that Sioc145 promotes insulin secretion in the amplifying pathway downstream of K ATp channels. In pancreatic islets, the treatment with Sioc145 enhances the second phase of insulin secretion. Increased insulin granules close to the plasma membrane are observed after Sioc145 treatment. Finally, the administration of Sioc145 to diabetic GK rats increases their serum insulin levels and improves glucose tolerance. Collectively, our studies identify Sioc145 as a novel glucose-dependent insulinotropic compound via selectively activating nPKCs.

  5. Aquasomes: A Self Assembling Nanobiopharmaceutical Carrier System for Bio-Active Molecules: A Review

    Patel JK

    2012-02-01

    Full Text Available Aquasomes are the self-assembling nanobiopharmaceutical carrier system, contains nanocrystallinecalcium phosphate or ceramic diamond, is covered by a glassy polyhydroxyl oligomeric film.Aquasomes are spherical (5–925nm particles used for drug and antigen delivery. Aquasomes are calledas “bodies of water" their water like properties protect and preserve fragile biological molecules. Its highdegree of surface exposure is used in targeting of bio-active molecules to specific sites. Three types ofcore materials are mainly used for producing aquasomes: Tin oxide, Nanocrystalline carbon ceramicsand Brushite. Calcium phosphate is the core of interest, due to its natural presence in the body. Thebrushite is unstable and converts to hydroxyapatite upon prolong storage and seems a better core for thepreparation of aquasomes. It is widely used for the preparation of implants. Aquasomes exploited as aRBC substitutes, vaccines for delivery of viral antigen and as targeted system for intracellular genetherapy. Enzyme activity and sensitivity towards molecular conformation made aquasome as a novelcarrier for enzymes like DNAses and pigment/dyes. This report reviews the principles of self assembly,the challenges of maintaining both the conformational integrity and biochemical activity of immobilizedsurface pairs.

  6. An active film-coating approach to enhance chemical stability of a potent drug molecule.

    Desai, Divyakant; Rao, Venkatramana; Guo, Hang; Li, Danping; Stein, Daniel; Hu, Frank Y; Kiesnowski, Chris

    2012-01-01

    Peliglitazar, a PPAR α/γ agonist, was found to undergo acid as well as base catalyzed degradation. The acid catalyzed degradation led to the formation of benzylic alcohol and glycine carbamate and the base catalyzed degradation led to formation of p-hydroxyanisole and an amine degradant. In capsule formulations, the capsules with the lowest drug-loading exhibited maximum instability even at 25 °C/60% RH storage condition. Incorporation of pH-modifiers to maintain 'micro-environmental pH' acidic did not prevent the formation of the base-catalyzed degradants. Traditional dry granulated tablet formulation which is qualitatively similar to the capsule formulations showed the presence of acid-catalyzed degradants even without the presence of an acidifying agent. On the other hand, traditional wet granulated tablet formulation showed mainly base-catalyzed degradants. Stability problems of the tablet formulation were aggravated because the potent molecule required low tablet strengths which resulted in low drug to excipient ratio. To stabilize the molecule, an active film-coating approach was explored. In this approach, the drug was sprayed with the coating material onto non-active containing tablet cores. This approach of trapping the drug particles into the coating material provided tablets with satisfactory chemical stability. The stability enhancement observed in the active coating approach is attributed to the higher drug to excipient ratio in the film coat of non-reactive coating material compared to that in the traditional dry or wet granulated formulations.

  7. Probe molecule studies: Active species in alcohol synthesis. Final report, July 1993--July 1994

    Blackmond, D.G.; Wender, I.; Oukaci, R.; Wang, Jian

    1994-07-01

    The objectives of this project are to investigate the role(s) of cobalt and copper in constructing the active sites for the formation of higher alcohols from CO/H{sub 2} over the Co-Cu based catalysts by using different reduction treatments and applying selected characterization tools such as TPR, TPD, XRD and XPS as well as to generate mechanistic information on the reaction pathway(s) and key intermediate(s) of higher alcohol synthesis from CO/H{sub 2} over Co-Cu/ZnO catalysts by the approach of in-situ addition of a probe molecule (nitromethane).

  8. Structure Based Discovery of Small Molecules to Regulate the Activity of Human Insulin Degrading Enzyme

    Bilal Çakir; Onur Dağliyan; Ezgi Dağyildiz; İbrahim Bariş; Ibrahim Halil Kavakli; Seda Kizilel; Metin Türkay

    2012-01-01

    Structure Based Discovery of Small Molecules to Regulate the Activity of Human Insulin Degrading Enzyme Bilal C¸ akir1, Onur Dag˘ liyan1, Ezgi Dag˘ yildiz1, I˙brahim Baris¸1, Ibrahim Halil Kavakli1,2*, Seda Kizilel1*, Metin Tu¨ rkay3* 1 Department of Chemical and Biological Engineering, Koc¸ University, Sariyer, Istanbul, Turkey, 2 Department of Molecular Biology and Genetics, Koc¸ University, Sariyer, Istanbul, Turkey, 3 Department of Industrial Engineering, Koc¸ University...

  9. Earthquake swarm activity in the Oaxaca segment of Middle American Subduction Zone

    Brudzinski, M. R.; Cabral, E.; Arciniega-Ceballos, A.

    2013-05-01

    An outstanding question in geophysics is the degree to which the newly discovered family of slow fault slip behaviors is related to more traditional earthquakes, especially since theoretical predictions indicate slip in the deeper transitional zone promotes failure in the shallower seismogenic zone. The Oaxacan segment of the Middle American Subduction zone is a natural region to pursue detailed studies of the spectrum of fault slip due to the unusually shallow subduction angle and short trench-to-coast distances that bring broad portions of the seismogenic and transitional zones of the plate interface inland. A deployment of broadband seismometers in this region has improved the network coverage to ~70 km station spacing since 2006, providing new opportunities to investigate smaller seismic phenomena. While characterization of tectonic tremor has been a prominent focus of this deployment, the improved network has also revealed productive earthquake swarms, whose sustained periods of similar magnitude earthquakes are also thought to be driven by slow slip. We identify a particularly productive earthquake swarm in July 2006 (~600 similar earthquakes detected), which occurred during a week-long episode of tectonic tremor and geodetically detected slow slip. Using a multi-station "template matching" waveform cross correlation technique, we have been able to detect and locate swarm earthquakes several orders of magnitude smaller than that of traditional processing, particularly during periods of increased background activity, because the detector is finely tuned to events with similar hypocentral location and focal mechanism. When we scan for repeats of the event families detected in the July 2006 sequence throughout the 6+ years since, we find these families were also activated during several other slow slip episodes, which indicates a link between slow slip in the transition zone and earthquakes at the downdip end of the seismogenic portion of the megathrust.

  10. Small molecule activators of SIRT1 replicate signaling pathways triggered by calorie restriction in vivo

    Lavu Siva

    2009-03-01

    Full Text Available Abstract Background Calorie restriction (CR produces a number of health benefits and ameliorates diseases of aging such as type 2 diabetes. The components of the pathways downstream of CR may provide intervention points for developing therapeutics for treating diseases of aging. The NAD+-dependent protein deacetylase SIRT1 has been implicated as one of the key downstream regulators of CR in yeast, rodents, and humans. Small molecule activators of SIRT1 have been identified that exhibit efficacy in animal models of diseases typically associated with aging including type 2 diabetes. To identify molecular processes induced in the liver of mice treated with two structurally distinct SIRT1 activators, SIRT501 (formulated resveratrol and SRT1720, for three days, we utilized a systems biology approach and applied Causal Network Modeling (CNM on gene expression data to elucidate downstream effects of SIRT1 activation. Results Here we demonstrate that SIRT1 activators recapitulate many of the molecular events downstream of CR in vivo, such as enhancing mitochondrial biogenesis, improving metabolic signaling pathways, and blunting pro-inflammatory pathways in mice fed a high fat, high calorie diet. Conclusion CNM of gene expression data from mice treated with SRT501 or SRT1720 in combination with supporting in vitro and in vivo data demonstrates that SRT501 and SRT1720 produce a signaling profile that mirrors CR, improves glucose and insulin homeostasis, and acts via SIRT1 activation in vivo. Taken together these results are encouraging regarding the use of small molecule activators of SIRT1 for therapeutic intervention into type 2 diabetes, a strategy which is currently being investigated in multiple clinical trials.

  11. ABOUT THE WAVE MECHANISM OF ACTIVATION OF FAULTS IN SEISMIC ZONES OF THE LITHOSPHERE IN MONGOLIA

    M. G. Mel’nikov

    2015-09-01

    Full Text Available The study is focused on earthquake migrations along active faults in seismic zones of Mongolia. The earthquake migrations are interpreted as a result of the influence of deformational waves. Vector velocities and other parameters of the deformational waves are studied. Based on data from largescale maps, local faults are compared, and differences and similarities of parameters of waves related to faults of different ranks are described.

  12. A Model of Ischemia-Induced Neuroblast Activation in the Adult Subventricular Zone

    Vergni, Davide; Castiglione, Filippo; Briani, Maya; Middei, Silvia; Alberdi, Elena; Reymann, Klaus G.; Natalini, Roberto; Volonté, Cinzia; Matute, Carlos; Cavaliere, Fabio

    2009-01-01

    We have developed a rat brain organotypic culture model, in which tissue slices contain cortex-subventricular zone-striatum regions, to model neuroblast activity in response to in vitro ischemia. Neuroblast activation has been described in terms of two main parameters, proliferation and migration from the subventricular zone into the injured cortex. We observed distinct phases of neuroblast activation as is known to occur after in vivo ischemia. Thus, immediately after oxygen/glucose deprivation (6–24 hours), neuroblasts reduce their proliferative and migratory activity, whereas, at longer time points after the insult (2 to 5 days), they start to proliferate and migrate into the damaged cortex. Antagonism of ionotropic receptors for extracellular ATP during and after the insult unmasks an early activation of neuroblasts in the subventricular zone, which responded with a rapid and intense migration of neuroblasts into the damaged cortex (within 24 hours). The process is further enhanced by elevating the production of the chemoattractant SDf-1α and may also be boosted by blocking the activation of microglia. This organotypic model which we have developed is an excellent in vitro system to study neurogenesis after ischemia and other neurodegenerative diseases. Its application has revealed a SOS response to oxygen/glucose deprivation, which is inhibited by unfavorable conditions due to the ischemic environment. Finally, experimental quantifications have allowed us to elaborate a mathematical model to describe neuroblast activation and to develop a computer simulation which should have promising applications for the screening of drug candidates for novel therapies of ischemia-related pathologies. PMID:19390597

  13. A model of ischemia-induced neuroblast activation in the adult subventricular zone.

    Davide Vergni

    Full Text Available We have developed a rat brain organotypic culture model, in which tissue slices contain cortex-subventricular zone-striatum regions, to model neuroblast activity in response to in vitro ischemia. Neuroblast activation has been described in terms of two main parameters, proliferation and migration from the subventricular zone into the injured cortex. We observed distinct phases of neuroblast activation as is known to occur after in vivo ischemia. Thus, immediately after oxygen/glucose deprivation (6-24 hours, neuroblasts reduce their proliferative and migratory activity, whereas, at longer time points after the insult (2 to 5 days, they start to proliferate and migrate into the damaged cortex. Antagonism of ionotropic receptors for extracellular ATP during and after the insult unmasks an early activation of neuroblasts in the subventricular zone, which responded with a rapid and intense migration of neuroblasts into the damaged cortex (within 24 hours. The process is further enhanced by elevating the production of the chemoattractant SDf-1alpha and may also be boosted by blocking the activation of microglia. This organotypic model which we have developed is an excellent in vitro system to study neurogenesis after ischemia and other neurodegenerative diseases. Its application has revealed a SOS response to oxygen/glucose deprivation, which is inhibited by unfavorable conditions due to the ischemic environment. Finally, experimental quantifications have allowed us to elaborate a mathematical model to describe neuroblast activation and to develop a computer simulation which should have promising applications for the screening of drug candidates for novel therapies of ischemia-related pathologies.

  14. Small-molecule CFTR activators increase tear secretion and prevent experimental dry eye disease.

    Flores, Alyssa M; Casey, Scott D; Felix, Christian M; Phuan, Puay W; Verkman, A S; Levin, Marc H

    2016-05-01

    Dry eye disorders, including Sjögren's syndrome, constitute a common problem in the aging population, with limited effective therapeutic options available. The cAMP-activated Cl(-) channel cystic fibrosis transmembrane conductance regulator (CFTR) is a major prosecretory channel at the ocular surface. We investigated whether compounds that target CFTR can correct the abnormal tear film in dry eye. Small-molecule activators of human wild-type CFTR identified by high-throughput screening were evaluated in cell culture and in vivo assays, to select compounds that stimulate Cl(-)-driven fluid secretion across the ocular surface in mice. An aminophenyl-1,3,5-triazine, CFTRact-K089, fully activated CFTR in cell cultures with EC50 ∼250 nM and produced an ∼8.5 mV hyperpolarization in ocular surface potential difference. When delivered topically, CFTRact-K089 doubled basal tear volume for 4 h and had no effect in CF mice. CFTRact-K089 showed sustained tear film bioavailability without detectable systemic absorption. In a mouse model of aqueous-deficient dry eye produced by lacrimal ablation, topical administration of 0.1 nmol CFTRact-K089 3 times daily restored tear volume to basal levels, preventing corneal epithelial disruption when initiated at the time of surgery and reversing it when started after development of dry eye. Our results support the potential utility of CFTR-targeted activators as a novel prosecretory treatment for dry eye.-Flores, A. M., Casey, S. D., Felix, C. M., Phuan, P. W., Verkman, A. S., Levin, M. H. Small-molecule CFTR activators increase tear secretion and prevent experimental dry eye disease.

  15. Influence of platelet activating factor on expression of adhesion molecules in experimental pancreatitis

    Hua Zhao; Ji-Wei Chen; Ya-Kui Zhou; Xue-Feng Zhou; Pei-Yun Li

    2003-01-01

    AIM: To determine whether Platelet activating factor (PAF)has a regulation role in the expression of adhesion moleculesand accumulation of neutrophils in a murine model of acutepancreatitis.METHODS: One hundred twenty-eight Kunming mice weredivided into four groups. Group 1 received 0.1 mi saline s.c.every hour for three hours (sham). Group 2 received cerulein(50 μg/kg dose s.c.) every hour for three hours. Group 3received AP and additional challenge of PAF (50 rg/kg inabsolute ethanol) (AP/PAF). Group 4 received AP, plustherapeutic treatment with GAB (25 mg dose i.p.) immediatelyafter the first challenge of cerulein (AP/GAB). Animals weresacrificed at 12 h after the first challenge of saline or cerulein.Adhesion molecules of pancreas were semi-quantified bySP methods. Standard assays were performed for serumamylase and myeloperoxidase activity (MPO) of pancreas.Histology of pancreas was scored in a blind manner. Watercontent of pancreas was also measured at the same time.RESULTS: Control pancreata showed negligible adhesionmolecule expression and neutrophil accumulation. Therewere evident adhesion molecules expression and neutrophilaccumulation in AP and AP/PAF compared with sham (P<0.05).AP/GAB had a lower level of adhesion molecules, neutrophils,and water content versus AP and AP/PAF (P<0.05). Histologyshowed a trend toward improvement in AP/GAB, but didnot reach statistical significance.CONCLUSION: PAF can induce the expression of adhesionmolecules that mediate neutrophil accumulation. The PAFantagonist reduces the expression of adhesion moleculesand the severity of inflammation when given immediatelyafter the induction of mild AP in mice. These results suggestthat PAF antagonism may be useful in the treatment of mildpancreatitis after its clinical onset.

  16. Conserved Active Site Residues Limit Inhibition of a Copper-Containing Nitrite By Small Molecules

    Tocheva, E.I.; Eltis, L.D.; Murphy, M.E.P.

    2009-05-26

    The interaction of copper-containing dissimilatory nitrite reductase from Alcaligenes faecalis S-6 ( AfNiR) with each of five small molecules was studied using crystallography and steady-state kinetics. Structural studies revealed that each small molecule interacted with the oxidized catalytic type 2 copper of AfNiR. Three small molecules (formate, acetate and nitrate) mimic the substrate by having at least two oxygen atoms for bidentate coordination to the type 2 copper atom. These three anions bound to the copper ion in the same asymmetric, bidentate manner as nitrite. Consistent with their weak inhibition of the enzyme ( K i >50 mM), the Cu-O distances in these AfNiR-inhibitor complexes were approximately 0.15 A longer than that observed in the AfNiR-nitrite complex. The binding mode of each inhibitor is determined in part by steric interactions with the side chain of active site residue Ile257. Moreover, the side chain of Asp98, a conserved residue that hydrogen bonds to type 2 copper-bound nitrite and nitric oxide, was either disordered or pointed away from the inhibitors. Acetate and formate inhibited AfNiR in a mixed fashion, consistent with the occurrence of second acetate binding site in the AfNiR-acetate complex that occludes access to the type 2 copper. A fourth small molecule, nitrous oxide, bound to the oxidized metal in a side-on fashion reminiscent of nitric oxide to the reduced copper. Nevertheless, nitrous oxide bound at a farther distance from the metal. The fifth small molecule, azide, inhibited the reduction of nitrite by AfNiR most strongly ( K ic = 2.0 +/- 0.1 mM). This ligand bound to the type 2 copper center end-on with a Cu-N c distance of approximately 2 A, and was the only inhibitor to form a hydrogen bond with Asp98. Overall, the data substantiate the roles of Asp98 and Ile257 in discriminating substrate from other small anions.

  17. Incorporation and characterization of biological molecules in droplet-interface bilayer networks for novel active systems

    Sarles, Stephen A.; Ghanbari Bavarsad, Pegah; Leo, Donald J.

    2009-03-01

    Biological molecules including phospholipids and proteins offer scientists and engineers a diverse selection of materials to develop new types of active materials and smart systems based on ion conduction. The inherent energy-coupling abilities of these components create novel kinds of transduction elements. Networks formed from droplet-interface bilayers (DIB) are a promising construct for creating cell mimics that allow for the assembly and study of these active biological molecules. The current-voltage relationship of symmetric, "lipid-in" dropletinterface bilayers are characterized using electrical impedance spectroscopy (EIS) and cyclic voltammetry (CV). "Lipid-in" diphytanoyl phosphatidylcholine (DPhPC) droplet-interface bilayers have specific resistances of nearly 10MΩ•cm2 and rupture at applied potentials greater than 300mV, indicating the "lipid-in" approach produces higher quality interfacial membranes than created using the original "lipid-out" method. The incorporation of phospholipids into the droplet interior allows for faster monolayer formation but does not inhibit the selfinsertion of transmembrane proteins into bilayer interfaces that separate adjacent droplets. Alamethicin proteins inserted into single and multi-DIB networks produce a voltage-dependent membrane conductance and current measurements on bilayers containing this type of protein exhibit a reversible, 3-4 order-of-magnitude conductance increase upon application of voltage.

  18. Expression of activated molecules on CD5(+)B lymphocytes in autoimmune hemolytic anemia.

    Zhu, Hongli; Xu, Wenyan; Liu, Hong; Wang, Huaquan; Fu, Rong; Wu, Yuhong; Qu, Wen; Wang, Guojin; Guan, Jing; Song, Jia; Xing, Limin; Shao, Zonghong

    2016-05-01

    To investigate the expression of activation molecules on CD5(+)B lymphocytes in peripheral blood of autoimmune hemolytic anemia (AIHA)/Evans patients. The expression of CD80, CD86, and CD69 on CD5(+)B lymphocytes was detected using flow cytometry in 30 AIHA/Evans patients, 18 normal controls (NC) and nine chronic lymphocytic leukemia (CLL) patients. CD80 on CD5(+)B lymphocytes in untreated patients was higher than that in remission patients (P 0.05), but lower than those of CD5(-)B lymphocytes in remission patients and NC (P 0.05). CD80 and CD86 on CD5(+)B lymphocytes was negatively correlated with hemoglobin (HB), C3, C4 (P < 0.05) and positively correlated with reticulocyte (Ret) (P < 0.05). CD69 on CD5(+) and CD5(-)B lymphocytes of CLL was higher than those of AIHA/Evans patients and NC (P < 0.05). The active molecules on CD5(+)B lymphocytes in peripheral blood of AIHA/Evans patients differ from those on CD5(-) and clonal CD5(+)B lymphocytes.

  19. Micro 3D ERT tomography for data assimilation modelling of active root zone

    Vanella, Daniela; Busato, Laura; Boaga, Jacopo; Cassiani, Giorgio; Binley, Andrew; Putti, Mario; Consoli, Simona

    2016-04-01

    Within the soil-plant-atmosphere system, root activity plays a fundamental role, as it connects different domains and allows a large part of the water and nutrient exchanges necessary for plant sustenance. The understanding of these processes is not only useful from an environmental point of view, making a fundamental contribution to the understanding of the critical zone dynamics, but also plays a pivotal role in precision agriculture, where the optimisation of water resources exploitation is mandatory and often carried out through deficit irrigation techniques. In this work, we present the results of non-invasive monitoring of the active root zone of two orange trees (Citrus sinensis, cv Tarocco Ippolito) located in an orange orchard in eastern Sicily (Italy) and drip irrigated with two different techniques: partial root drying and 100% crop evapotranspiration. The main goal of the monitoring activity is to assess possible differences between the developed root systems and the root water uptake between the two irrigation strategies. The monitoring is conducted using 3D micro-electrical resistivity tomography (ERT) based on an apparatus composed of a number of micro-boreholes (about 1.2 m deep) housing 12 electrodes each, plus a number of surface electrodes. Time-lapse measurements conducted both with long-term periodicity and short-term repetition before and after irrigation clearly highlight the presence and distribution of root water uptake zone both at shallow and larger depth, likely to correspond to zones utilized during the irrigation period (shallow) and during the time when the crop is not irrigated (deep). Subsidiary information is available in terms of precipitation, sap flow measurements and micrometeorological evapotranspiration estimates. This data ensemble lends itself to the assimilation into a variably saturated flow model, where both soil hydraulic parameters and root distribution shall be identified. Preliminary results in this directions show

  20. p53 Small Molecule Inhibitor Enhances Temozolomide Cytotoxic Activity against Intracranial Glioblastoma Xenografts

    Dinca, Eduard B.; Lu, Kan V.; Sarkaria, Jann N.; Pieper, Russell O.; Prados, Michael D.; Haas-Kogan, Daphne A.; VandenBerg, Scott R.; Berger, Mitchel S.; James, C. David

    2010-01-01

    In this study we investigated corresponding precursor and active forms of a p53 small molecule inhibitor for effect on temozolomide (TMZ) anti-tumor activity against glioblastoma (GBM), using both in vitro and in vivo experimental approaches. Results from in vitro cell viability analysis showed that the cytotoxic activity of TMZ was substantially increased when GBMs with wild-type p53 were co-treated with the active form of p53 inhibitor, and this heightened cytotoxic response was accompanied by increased PARP cleavage as well as elevated cellular phospho-H2AX. Analysis of the same series of GBMs, as intracranial xenografts in athymic mice, and administering corresponding p53 inhibitor precursor, that is converted to the active compound in vivo, yielded results consistent with the in vitro analyses: i.e., TMZ + p53 inhibitor precursor co-treatment, of three distinct wild-type p53 GBM xenografts, resulted in significant enhancement of TMZ anti-tumor effect relative to treatment with TMZ alone, as indicated by serial bioluminescence monitoring as well as survival analysis (p < 0.001 for co-treatment survival benefit in each case). Mice receiving intracranial injection with p53 null GBM showed similar survival benefit from TMZ treatment regardless of the presence or absence of p53 inhibitor precursor. In total, our results indicate that the p53 active and precursor inhibitor pair enhance TMZ cytotoxicity in vitro and in vivo, respectively, and do so in a p53-dependent manner. PMID:19074867

  1. Small-Molecule Inhibition and Activation-Loop Trans-Phosphorylation of the IGF1 Receptor

    Wu,J.; Li, W.; Craddock, B.; Foreman, K.; Mulvihill, M.; Ji, Q.; Miller, W.; Hubbard, S.

    2008-01-01

    The insulin-like growth factor-1 receptor (IGF1R) is a receptor tyrosine kinase (RTK) that has a critical role in mitogenic signalling during embryogenesis and an antiapoptotic role in the survival and progression of many human tumours. Here, we present the crystal structure of the tyrosine kinase domain of IGF1R (IGF1RK), in its unphosphorylated state, in complex with a novel compound, cis-3-[3-(4-methyl-piperazin-l-yl)-cyclobutyl]-1-(2-phenyl-quinolin-7-yl)-imidazo[1, 5-a]pyrazin-8-ylamine (PQIP), which we show is a potent inhibitor of both the unphosphorylated (basal) and phosphorylated (activated) states of the kinase. PQIP interacts with residues in the ATP-binding pocket and in the activation loop, which confers specificity for IGF1RK and the highly related insulin receptor (IR) kinase. In this crystal structure, the IGF1RK active site is occupied by Tyr1135 from the activation loop of an symmetry (two-fold)-related molecule. This dimeric arrangement affords, for the first time, a visualization of the initial trans-phosphorylation event in the activation loop of an RTK, and provides a molecular rationale for a naturally occurring mutation in the activation loop of the IR that causes type II diabetes mellitus.

  2. Discovery of novel small molecule activators of β-catenin signaling.

    Folkert Verkaar

    Full Text Available Wnt/β-catenin signaling plays a major role in embryonic development and adult stem cell maintenance. Reduced activation of the Wnt/β-catenin pathway underlies neurodegenerative disorders and aberrations in bone formation. Screening of a small molecule compound library with a β-galactosidase fragment complementation assay measuring β-catenin nuclear entry revealed bona fide activators of β-catenin signaling. The compounds stabilized cytoplasmic β-catenin and activated β-catenin-dependent reporter gene activity. Although the mechanism through which the compounds activate β-catenin signaling has yet to be determined, several key regulators of Wnt/β-catenin signaling, including glycogen synthase kinase 3 and Frizzled receptors, were excluded as the molecular target. The compounds displayed remarkable selectivity, as they only induced β-catenin signaling in a human osteosarcoma U2OS cell line and not in a variety of other cell lines examined. Our data indicate that differences in cellular Wnt/β-catenin signaling machinery can be exploited to identify cell type-specific activators of Wnt/β-catenin signaling.

  3. Novel small-molecule AMPK activator orally exerts beneficial effects on diabetic db/db mice

    Li, Yuan-Yuan; Yu, Li-Fang; Zhang, Li-Na; Qiu, Bei-Ying; Su, Ming-Bo; Wu, Fang; Chen, Da-Kai; Pang, Tao; Gu, Min; Zhang, Wei; Ma, Wei-Ping; Jiang, Hao-Wen; Li, Jing-Ya, E-mail: jyli@mail.shcnc.ac.cn; Nan, Fa-Jun, E-mail: fjnan@mail.shcnc.ac.cn; Li, Jia, E-mail: jli@mail.shcnc.ac.cn

    2013-12-01

    AMP-activated protein kinase (AMPK), which is a pivotal guardian of whole-body energy metabolism, has become an attractive therapeutic target for metabolic syndrome. Previously, using a homogeneous scintillation proximity assay, we identified the small-molecule AMPK activator C24 from an optimization based on the original allosteric activator PT1. In this paper, the AMPK activation mechanism of C24 and its potential beneficial effects on glucose and lipid metabolism on db/db mice were investigated. C24 allosterically stimulated inactive AMPK α subunit truncations and activated AMPK heterotrimers by antagonizing autoinhibition. In primary hepatocytes, C24 increased the phosphorylation of AMPK downstream target acetyl-CoA carboxylase dose-dependently without changing intracellular AMP/ATP ratio, indicating its allosteric activation in cells. Through activating AMPK, C24 decreased glucose output by down-regulating mRNA levels of phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase) in primary hepatocytes. C24 also decreased the triglyceride and cholesterol contents in HepG2 cells. Due to its improved bioavailability, chronic oral treatment with multiple doses of C24 significantly reduced blood glucose and lipid levels in plasma, and improved the glucose tolerance of diabetic db/db mice. The hepatic transcriptional levels of PEPCK and G6Pase were reduced. These results demonstrate that this orally effective activator of AMPK represents a novel approach to the treatment of metabolic syndrome. - Highlights: • C24 activates AMPK through antagonizing autoinhibition within α subunit. • C24 activates AMPK in hepatocytes and decreases glucose output via AMPK. • C24 exerts beneficial effects on diabetic db/db mice. • C24 represents a novel therapeutic for treatment of metabolic syndrome.

  4. VAMP4 Is an Essential Cargo Molecule for Activity-Dependent Bulk Endocytosis.

    Nicholson-Fish, Jessica C; Kokotos, Alexandros C; Gillingwater, Thomas H; Smillie, Karen J; Cousin, Michael A

    2015-12-02

    The accurate formation of synaptic vesicles (SVs) and incorporation of their protein cargo during endocytosis is critical for the maintenance of neurotransmission. During intense neuronal activity, a transient and acute accumulation of SV cargo occurs at the plasma membrane. Activity-dependent bulk endocytosis (ADBE) is the dominant SV endocytosis mode under these conditions; however, it is currently unknown how ADBE mediates cargo retrieval. We examined the retrieval of different SV cargo molecules during intense stimulation using a series of genetically encoded pH-sensitive reporters in neuronal cultures. The retrieval of only one reporter, VAMP4-pHluorin, was perturbed by inhibiting ADBE. This selective recovery was confirmed by the enrichment of endogenous VAMP4 in purified bulk endosomes formed by ADBE. VAMP4 was also essential for ADBE, with a cytoplasmic di-leucine motif being critical for this role. Therefore, VAMP4 is the first identified ADBE cargo and is essential for this endocytosis mode to proceed.

  5. Small Molecule-Induced Allosteric Activation of the Vibrio Cholerae RTX Cysteine Protease Domain

    Lupardus, P.J.; Shen, A.; Bogyo, M.; Garcia, K.C.

    2009-05-19

    Vibrio cholerae RTX (repeats in toxin) is an actin-disrupting toxin that is autoprocessed by an internal cysteine protease domain (CPD). The RTX CPD is efficiently activated by the eukaryote-specific small molecule inositol hexakisphosphate (InsP{sub 6}), and we present the 2.1 angstrom structure of the RTX CPD in complex with InsP{sub 6}. InsP{sub 6} binds to a conserved basic cleft that is distant from the protease active site. Biochemical and kinetic analyses of CPD mutants indicate that InsP{sub 6} binding induces an allosteric switch that leads to the autoprocessing and intracellular release of toxin-effector domains.

  6. Periodic activity of secretory glands of stomach in ulcer erosion of gastro-duodenal zone

    A. I. Rudenko

    2005-12-01

    Full Text Available It was fixed, that development of atophanum-carbacholimun ulcer of the gastroduodenal zone invoked various changes of secretory activity of the stomach. The changes directly depend on a progress of pathological process. As this takes place the reaction of stomach secretory glands varies under the stimulation with histamine: the decrease of stomach secretory glands’ work capacity till 10th day and its increase after 10–15th day were observed. Disorders of the glands’ ultradian rhythms at initial stages of modeling of gastrointestinal nervous regulation disturbances testify to dependence of periodic activity of gastrointestinal tract on resistance of regulatory mechanisms correlation.

  7. The energy balance and pressure in the solar transition zone for network and active region features

    Nicolas, K. R.; Bartoe, J.-D. F.; Brueckner, G. E.; Vanhoosier, M. E.

    1979-01-01

    The electron pressure and energy balance in the solar transition zone are determined for about 125 network and active region features on the basis of high spectral and spatial resolution extreme ultraviolet spectra. Si III line intensity ratios obtained from the Naval Research Laboratory high-resolution telescope and spectrograph during a rocket flight are used as diagnostics of electron density and pressure for solar features near 3.5 x 10 to the 4th K. Observed ratios are compared with the calculated dependence of the 1301 A/1312 A and 1301 A/1296 A line intensity ratios on electron density, temperature and pressure. Electron densities ranging from 2 x 10 to the 10th/cu cm to 10 to the 12th/cu cm and active region pressures from 3 x 10 to the 15th to 10 to the 16th/cu cm K are obtained. Energy balance calculations reveal the balance of the divergence of the conductive flux and turbulent energy dissipation by radiative energy losses in a plane-parallel homogeneous transition zone (fill factor of 1), and an energy source requirement for a cylindrical zone geometry (fill factor less than 0.04).

  8. 78 FR 7394 - Foreign-Trade Zone 41-Milwaukee, WI; Notification of Proposed Production Activity; CNH America...

    2013-02-01

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF COMMERCE Foreign-Trade Zones Board Foreign-Trade Zone 41--Milwaukee, WI; Notification of Proposed Production Activity; CNH America, LLC, Subzone 41I (Tractors and Tractor/Combine Components), Racine, WI The Port...

  9. 78 FR 54234 - Foreign-Trade Zone 26-Atlanta, Georgia, Authorization of Production Activity PBR, Inc. d/b/a...

    2013-09-03

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF COMMERCE Foreign-Trade Zones Board Foreign-Trade Zone 26--Atlanta, Georgia, Authorization of Production Activity PBR, Inc. d/b/a SKAPS Industries (Polypropylene Geotextiles), Athens, Georgia On April 8,...

  10. 77 FR 46377 - Foreign-Trade Zone 37-Orange County, NY, Notification of Proposed Production Activity, Takasago...

    2012-08-03

    ..., mixtures of odiferous substances and essential oils (duty rate ranges from duty free to 7%). Public comment... Foreign-Trade Zones Board Foreign-Trade Zone 37--Orange County, NY, Notification of Proposed Production... (Takasago) submitted a notification of proposed production activity for their facility located in...

  11. Stellar activity mimics a habitable-zone planet around Kapteyn's star

    Robertson, Paul; Mahadevan, Suvrath

    2015-01-01

    Kapteyn's star is an old M subdwarf believed to be a member of the Galactic halo population of stars. A recent study has claimed the existence of two super-Earth planets around the star based on radial velocity (RV) observations. The innermost of these candidate planets--Kapteyn b (P = 48 days)--resides within the circumstellar habitable zone. Given recent progress in understanding the impact of stellar activity in detecting planetary signals, we have analyzed the observed HARPS data for signatures of stellar activity. We find that while Kapteyn's star is photometrically very stable, a suite of spectral activity indices reveals a large-amplitude rotation signal, and we determine the stellar rotation period to be 143 days. The spectral activity tracers are strongly correlated with the purported RV signal of "planet b," and the 48-day period is an integer fraction (1/3) of the stellar rotation period. We conclude that Kapteyn b is not a planet in the Habitable Zone, but an artifact of stellar activity.

  12. More Active Living-oriented County and Municipal Zoning is Associated with Increased Adult Leisure Time Physical Activity-United States, 2011.

    Chriqui, Jamie F; Nicholson, Lisa M; Thrun, Emily; Leider, Julien; Slater, Sandy J

    2016-01-01

    Although zoning is recognized for its role in facilitating healthy communities, no study has examined whether active living-oriented zoning codes are associated with adult leisure time physical activity (PA). This study sought to fill this gap and hypothesized that adult leisure time PA would be greater in communities with more progressive zoning code reforms and more active living-oriented zoning. Zoning codes for 1,617 county and municipal jurisdictions located in 30 states (covering ~40% of the U.S. population) were evaluated for code reform zoning and 11 active living markers. County-aggregated zoning measures were created for linking with five adult PA behaviors obtained from the 2011 Behavioral Risk Factor Surveillance System controlling for individual and county sociodemographics. Zoning elements most associated with adult PA included requirements for mixed use, active and passive recreation, bike parking/street furniture, and bike-pedestrian trails/paths. This study provides new insights as to the role that zoning can play in facilitating adult PA.

  13. Peptidomimetic Small Molecules Disrupt Type IV Secretion System Activity in Diverse Bacterial Pathogens

    Carrie L. Shaffer

    2016-04-01

    Full Text Available Bacteria utilize complex type IV secretion systems (T4SSs to translocate diverse effector proteins or DNA into target cells. Despite the importance of T4SSs in bacterial pathogenesis, the mechanism by which these translocation machineries deliver cargo across the bacterial envelope remains poorly understood, and very few studies have investigated the use of synthetic molecules to disrupt T4SS-mediated transport. Here, we describe two synthetic small molecules (C10 and KSK85 that disrupt T4SS-dependent processes in multiple bacterial pathogens. Helicobacter pylori exploits a pilus appendage associated with the cag T4SS to inject an oncogenic effector protein (CagA and peptidoglycan into gastric epithelial cells. In H. pylori, KSK85 impedes biogenesis of the pilus appendage associated with the cag T4SS, while C10 disrupts cag T4SS activity without perturbing pilus assembly. In addition to the effects in H. pylori, we demonstrate that these compounds disrupt interbacterial DNA transfer by conjugative T4SSs in Escherichia coli and impede vir T4SS-mediated DNA delivery by Agrobacterium tumefaciens in a plant model of infection. Of note, C10 effectively disarmed dissemination of a derepressed IncF plasmid into a recipient bacterial population, thus demonstrating the potential of these compounds in mitigating the spread of antibiotic resistance determinants driven by conjugation. To our knowledge, this study is the first report of synthetic small molecules that impair delivery of both effector protein and DNA cargos by diverse T4SSs.

  14. Mytilus galloprovincialis myticin C: a chemotactic molecule with antiviral activity and immunoregulatory properties.

    Balseiro, Pablo; Falcó, Alberto; Romero, Alejandro; Dios, Sonia; Martínez-López, Alicia; Figueras, Antonio; Estepa, Amparo; Novoa, Beatriz

    2011-01-01

    Previous research has shown that an antimicrobial peptide (AMP) of the myticin class C (Myt C) is the most abundantly expressed gene in cDNA and suppressive subtractive hybridization (SSH) libraries after immune stimulation of mussel Mytilus galloprovincialis. However, to date, the expression pattern, the antimicrobial activities and the immunomodulatory properties of the Myt C peptide have not been determined. In contrast, it is known that Myt C mRNA presents an unusual and high level of polymorphism of unidentified biological significance. Therefore, to provide a better understanding of the features of this interesting molecule, we have investigated its function using four different cloned and expressed variants of Myt C cDNA and polyclonal anti-Myt C sera. The in vivo results suggest that this AMP, mainly present in hemocytes, could be acting as an immune system modulator molecule because its overexpression was able to alter the expression of mussel immune-related genes (as the antimicrobial peptides Myticin B and Mytilin B, the C1q domain-containing protein MgC1q, and lysozyme). Moreover, the in vitro results indicate that Myt C peptides have antimicrobial and chemotactic properties. Their recombinant expression in a fish cell line conferred protection against two different fish viruses (enveloped and non-enveloped). Cell extracts from Myt C expressing fish cells were also able to attract hemocytes. All together, these results suggest that Myt C should be considered not only as an AMP but also as the first chemokine/cytokine-like molecule identified in bivalves and one of the few examples in all of the invertebrates.

  15. Mytilus galloprovincialis myticin C: a chemotactic molecule with antiviral activity and immunoregulatory properties.

    Pablo Balseiro

    Full Text Available Previous research has shown that an antimicrobial peptide (AMP of the myticin class C (Myt C is the most abundantly expressed gene in cDNA and suppressive subtractive hybridization (SSH libraries after immune stimulation of mussel Mytilus galloprovincialis. However, to date, the expression pattern, the antimicrobial activities and the immunomodulatory properties of the Myt C peptide have not been determined. In contrast, it is known that Myt C mRNA presents an unusual and high level of polymorphism of unidentified biological significance. Therefore, to provide a better understanding of the features of this interesting molecule, we have investigated its function using four different cloned and expressed variants of Myt C cDNA and polyclonal anti-Myt C sera. The in vivo results suggest that this AMP, mainly present in hemocytes, could be acting as an immune system modulator molecule because its overexpression was able to alter the expression of mussel immune-related genes (as the antimicrobial peptides Myticin B and Mytilin B, the C1q domain-containing protein MgC1q, and lysozyme. Moreover, the in vitro results indicate that Myt C peptides have antimicrobial and chemotactic properties. Their recombinant expression in a fish cell line conferred protection against two different fish viruses (enveloped and non-enveloped. Cell extracts from Myt C expressing fish cells were also able to attract hemocytes. All together, these results suggest that Myt C should be considered not only as an AMP but also as the first chemokine/cytokine-like molecule identified in bivalves and one of the few examples in all of the invertebrates.

  16. Screening of pharmacologically active small molecule compounds identifies antifungal agents against Candida biofilms

    Takao eWatamoto

    2015-12-01

    Full Text Available Candida species have emerged as important and common opportunistic human pathogens, particularly in immunocompromised individuals. The current antifungal therapies either have toxic side effects or are insufficiently effect. The aim of this study is develop new small-molecule antifungal compounds by library screening methods using C. albicans, and to evaluate their antifungal effects on Candida biofilms and cytotoxic effects on human cells. Wild-type C. albicans strain SC5314 was used in library screening. To identify antifungal compounds, we screened a small-molecule library of 1,280 pharmacologically active compounds (LOPAC1280TM using an antifungal susceptibility test (AST. To investigate the antifungal effects of the hit compounds, ASTs were conducted using Candida strains in various growth modes, including biofilms. We tested the cytotoxicity of the hit compounds using human gingival fibroblast (hGF cells to evaluate their clinical safety. Only 35 compounds were identified by screening, which inhibited the metabolic activity of C. albicans by >50%. Of these, 26 compounds had fungistatic effects and 9 compounds had fungicidal effects on C. albicans. Five compounds, BAY11-7082, BAY11-7085, sanguinarine chloride hydrate, ellipticine and CV-3988, had strong fungicidal effects and could inhibit the metabolic activity of Candida biofilms. However, BAY11-7082, BAY11-7085, sanguinarine chloride hydrate and ellipticine were cytotoxic to hGF cells at low concentrations. CV-3988 showed no cytotoxicity at a fungicidal concentration.Four of the compounds identified, BAY11-7082, BAY11-7085, sanguinarine chloride hydrate and ellipticine, had toxic effects on Candida strains and hGF cells. In contrast, CV-3988 had fungicidal effects on Candida strains, but low cytotoxic effects on hGF cells. Therefore, this screening reveals agent, CV-3988 that was previously unknown to be antifungal agent, which could be a novel therapies for superficial mucosal

  17. Dendrimers and Polyamino-Phenolic Ligands: Activity of New Molecules Against Legionella pneumophila Biofilms

    Andreozzi, Elisa; Barbieri, Federica; Ottaviani, Maria F.; Giorgi, Luca; Bruscolini, Francesca; Manti, Anita; Battistelli, Michela; Sabatini, Luigia; Pianetti, Anna

    2016-01-01

    Legionnaires’ disease is a potentially fatal pneumonia caused by Legionella pneumophila, an aquatic bacterium often found within the biofilm niche. In man-made water systems microbial biofilms increase the resistance of legionella to disinfection, posing a significant threat to public health. Disinfection methods currently used in water systems have been shown to be ineffective against legionella over the long-term, allowing recolonization by the biofilm-protected microorganisms. In this study, the anti-biofilm activity of previously fabricated polyamino-phenolic ligands and polyamidoamine dendrimers was investigated against legionella mono-species and multi-species biofilms formed by L. pneumophila in association with other bacteria that can be found in tap water (Aeromonas hydrophila, Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae). Bacterial ability to form biofilms was verified using a crystal violet colorimetric assay and testing cell viability by real-time quantitative PCR and Plate Count assay. The concentration of the chemicals tested as anti-biofilm agents was chosen based on cytotoxicity assays: the highest non-cytotoxic chemical concentration was used for biofilm inhibition assays, with dendrimer concentration 10-fold higher than polyamino-phenolic ligands. While Macrophen and Double Macrophen were the most active substances among polyamino-phenolic ligands, dendrimers were overall twofold more effective than all other compounds with a reduction up to 85 and 73% of legionella and multi-species biofilms, respectively. Chemical interaction with matrix molecules is hypothesized, based on SEM images and considering the low or absent anti-microbial activity on planktonic bacteria showed by flow cytometry. These data suggest that the studied compounds, especially dendrimers, could be considered as novel molecules in the design of research projects aimed at the development of efficacious anti-biofilm disinfection treatments of water systems

  18. Dendrimers and polyamino-phenolic ligands: activity of new molecules against Legionella pneumophila biofilms.

    Elisa eAndreozzi

    2016-03-01

    Full Text Available Legionnaires’ disease is a potentially fatal pneumonia caused by Legionella pneumophila, an aquatic bacterium often found within the biofilm niche. In man-made water systems microbial biofilms increase the resistance of legionella to disinfection, posing a significant threat to public health. Disinfection methods currently used in water systems have been shown to be ineffective against legionella over the long-term, allowing recolonization by the biofilm-protected microorganisms. In this study, the anti-biofilm activity of previously fabricated polyamino-phenolic ligands and polyamidoamine dendrimers was investigated against legionella mono-species and multi-species biofilms formed by L. pneumophila in association with other bacteria that can be found in tap water (Aeromonas hydrophila, Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae. Bacterial ability to form biofilms was verified using a crystal violet colorimetric assay and testing cell viability by real-time quantitative PCR and Plate Count assay. The concentration of the chemicals tested as anti-biofilm agents was chosen based on cytotoxicity assays: the highest non-cytotoxic chemical concentration was used for biofilm inhibition assays, with dendrimer concentration ten-fold higher than polyamino-phenolic ligands. While Macrophen and Double Macrophen were the most active substances among polyamino-phenolic ligands, dendrimers were overall two-fold more effective than all other compounds with a reduction up to 85% and 73% of legionella and multi-species biofilms, respectively. Chemical interaction with matrix molecules is hypothesized, based on SEM images and considering the low or absent anti-microbial activity on planktonic bacteria showed by flow cytometry. These data suggest that the studied compounds, especially dendrimers, could be considered as novel molecules in the design of research projects aimed at the development of efficacious anti-biofilm disinfection

  19. Landform development in a zone of active Gedi Fault, Eastern Kachchh rift basin, India

    Kothyari, Girish Ch.; Rastogi, B. K.; Morthekai, P.; Dumka, Rakesh K.

    2016-02-01

    An earthquake of 2006 Mw 5.7 occurred along east-west trending Gedi Fault (GF) to the north of the Kachchh rift basin in western India which had the epicenter in the Wagad upland, which is approximately 60 km northeast of the 2001 Mw 7.7 earthquake site (or epicenter). Development of an active fault scarp, shifting of a river channel, offsetting of streams and uplift of the ground indicate that the terrain is undergoing active deformation. Based on detailed field investigations, three major faults that control uplifts have been identified in the GF zone. These uplifts were developed in a step-over zone of the GF, and formed due to compressive force generated by left-lateral motion within the segmented blocks. In the present research, a terrace sequence along the north flowing Karaswali river in a tectonically active GF zone has been investigated. Reconstructions based on geomorphology and terrace stratigraphy supported by optical chronology suggest that the fluvial aggradation in the Wagad area was initiated during the strengthening (at ~ 8 ka) and declining (~ 4 ka) of the Indian Summer Monsoon (ISM). The presence of younger valley fill sediments which are dated ~ 1 ka is ascribed to a short lived phase of renewed strengthening of ISM before present day aridity. Based on terrace morphology two major phases of enhanced uplift have been estimated. The older uplift event dated to 8 ka is represented by the Tertiary bedrock surfaces which accommodated the onset of valley-fill aggradation. The younger event of enhanced uplift dated to 4 ka was responsible for the incision of the older valley fill sediments and the Tertiary bedrock. These ages suggest that the average rate of uplift ranges from 0.3 to 1.1 mm/yr during the last 9 ka implying active nature of the area.

  20. A Pipeline for Screening Small Molecules with Growth Inhibitory Activity against Burkholderia cenocepacia.

    Carrie Selin

    Full Text Available Infections with the bacteria Burkholderia cepacia complex (Bcc are very difficult to eradicate in cystic fibrosis patients due the intrinsic resistance of Bcc to most available antibiotics and the emergence of multiple antibiotic resistant strains during antibiotic treatment. In this work, we used a whole-cell based assay to screen a diverse collection of small molecules for growth inhibitors of a relevant strain of Bcc, B. cenocepacia K56-2. The primary screen used bacterial growth in 96-well plate format and identified 206 primary actives among 30,259 compounds. From 100 compounds with no previous record of antibacterial activity secondary screening and data mining selected a total of Bce bioactives that were further analyzed. An experimental pipeline, evaluating in vitro antibacterial and antibiofilm activity, toxicity and in vivo antibacterial activity using C. elegans was used for prioritizing compounds with better chances to be further investigated as potential Bcc antibacterial drugs. This high throughput screen, along with the in vitro and in vivo analysis highlights the utility of this experimental method to quickly identify bioactives as a starting point of antibacterial drug discovery.

  1. Dissecting allosteric effects of activator-coactivator complexes using a covalent small molecule ligand.

    Wang, Ningkun; Lodge, Jean M; Fierke, Carol A; Mapp, Anna K

    2014-08-19

    Allosteric binding events play a critical role in the formation and stability of transcriptional activator-coactivator complexes, perhaps in part due to the often intrinsically disordered nature of one or more of the constituent partners. The kinase-inducible domain interacting (KIX) domain of the master coactivator CREB binding protein/p300 is a conformationally dynamic domain that complexes with transcriptional activators at two discrete binding sites in allosteric communication. The complexation of KIX with the transcriptional activation domain of mixed-lineage leukemia protein leads to an enhancement of binding by the activation domain of CREB (phosphorylated kinase-inducible domain of CREB) to the second site. A transient kinetic analysis of the ternary complex formation aided by small molecule ligands that induce positive or negative cooperative binding reveals that positive cooperativity is largely governed by stabilization of the bound complex as indicated by a decrease in koff. Thus, this suggests the increased binding affinity for the second ligand is not due to an allosteric creation of a more favorable binding interface by the first ligand. This is consistent with data from us and from others indicating that the on rates of conformationally dynamic proteins approach the limits of diffusion. In contrast, negative cooperativity is manifested by alterations in both kon and koff, suggesting stabilization of the binary complex.

  2. Topical Anti-inflammatory Activity of New Hybrid Molecules of Terpenes and Synthetic Drugs.

    Theoduloz, Cristina; Delporte, Carla; Valenzuela-Barra, Gabriela; Silva, Ximena; Cádiz, Solange; Bustamante, Fernanda; Pertino, Mariano Walter; Schmeda-Hirschmann, Guillermo

    2015-06-18

    The aim of the study was to assess changes in the activity of anti-inflammatory terpenes from Chilean medicinal plants after the formation of derivatives incorporating synthetic anti-inflammatory agents. Ten new hybrid molecules were synthesized combining terpenes (ferruginol (1), imbricatolic acid (2) and oleanolic acid (3)) with ibuprofen (4) or naproxen (5). The topical anti-inflammatory activity of the compounds was assessed in mice by the arachidonic acid (AA) and 12-O-tetradecanoyl phorbol 13-acetate (TPA) induced ear edema assays. Basal cytotoxicity was determined towards human lung fibroblasts, gastric epithelial cells and hepatocytes. At 1.4 µmol/mouse, a strong anti-inflammatory effect in the TPA assay was observed for oleanoyl ibuprofenate 12 (79.9%) and oleanoyl ibuprofenate methyl ester 15 (80.0%). In the AA assay, the best activity was observed for 12 at 3.2 µmol/mouse, with 56.8% reduction of inflammation, in the same range as nimesulide (48.9%). All the terpenyl-synthetic anti-inflammatory hybrids showed better effects in the TPA assay, with best activity for 6, 12 and 15. The cytotoxicity of the compounds 8 and 10 with a free COOH, was higher than that of 2. The derivatives from 3 were less toxic than the triterpene. Several of the new compounds presented better anti-inflammatory effect and lower cytotoxicity than the parent terpenes.

  3. A Preliminary Study on the New Activity Features of the Lajishan Mountain Fault Zone in Qinghai Province

    Yuan Daoyang; Zhang Peizhen; Lei Zhongsheng; Liu Baichi; Liu Xiaolong

    2005-01-01

    The Lajishan Mountain fault zone consists of two NE-protruding arcuate faults, i.e. the northern and southern margin fault of Lajishan Mountain with the fault length of 230km and 220km respectively. The fault zone is located in the large-scale compressional structure zone and tectonic gradient zone in-between the NNW-trending right-lateral strike-slip ReshuiRiyueshan fault zone and the NWW-trending left-lateral strike-slip northern margin of west Qinling Ranges fault zone is also an important boundary fault zone, separating the XiningMinhe basin and the Xunhua-Hualong basin at the southern and northern sides of the Lajishan Mountain respectively. Geologic geomorphic evidences of new activity revealed by field investigations indicate that the latest movement of the Lajishan fault zone was in late Epipleistocene (only a few segments were active in early Holocene ) and is mainly of compressive thrusting with slightly left-lateral strike-slip component. The above movement has possibly resulted in the occurrence of about 20 moderate earthquakes of magnitude around 5.0.The Lajishan region can therefore be regarded as a seismotectonic window to reflect tectonic movement and earthquake activity.

  4. DEFORMATION WAVES AS A TRIGGER MECHANISM OF SEISMIC ACTIVITY IN SEISMIC ZONES OF THE CONTINENTAL LITHOSPHERE

    S. I. Sherman

    2015-09-01

    Full Text Available Deformation waves as a trigger mechanism of seismic activity and migration of earthquake foci have been under discussion by researchers in seismology and geodynamics for over 50 years. Four sections of this article present available principal data on impacts of wave processes on seismicity and new data. The first section reviews analytical and experimental studies aimed at identification of relationships between wave processes in the lithosphere and seismic activity manifested as space-and-time migration of individual earthquake foci or clusters of earthquakes. It is concluded that with a systematic approach, instead of using a variety of terms to denote waves that trigger seismic process in the lithosphere, it is reasonable to apply the concise definition of ‘deformation waves’, which is most often used in fact.The second section contains a description of deformation waves considered as the trigger mechanism of seismic activity. It is concluded that a variety of methods are applied to identify deformation waves, and such methods are based on various research methods and concepts that naturally differ in sensitivity concerning detection of waves and/or impact of the waves on seismic process. Epicenters of strong earthquakes are grouped into specific linear or arc-shaped systems, which common criterion is the same time interval of the occurrence of events under analysis. On site the systems compose zones with similar time sequences, which correspond to the physical notion of moving waves (Fig. 9. Periods of manifestation of such waves are estimated as millions of years, and a direct consideration of the presence of waves and wave parameters is highly challenging. In the current state-of-the-art, geodynamics and seismology cannot provide any other solution yet.The third section presents a solution considering record of deformation waves in the lithosphere. With account of the fact that all the earthquakes with М≥3.0 are associated with

  5. Active Crustal Faults in the Forearc Region, Guerrero Sector of the Mexican Subduction Zone

    Gaidzik, Krzysztof; Ramírez-Herrera, Maria Teresa; Kostoglodov, Vladimir

    2016-10-01

    This work explores the characteristics and the seismogenic potential of crustal faults on the overriding plate in an area of high seismic hazard associated with the occurrence of subduction earthquakes and shallow earthquakes of the overriding plate. We present the results of geomorphic, structural, and fault kinematic analyses conducted on the convergent margin between the Cocos plate and the forearc region of the overriding North American plate, within the Guerrero sector of the Mexican subduction zone. We aim to determine the active tectonic processes in the forearc region of the subduction zone, using the river network pattern, topography, and structural data. We suggest that in the studied forearc region, both strike-slip and normal crustal faults sub-parallel to the subduction zone show evidence of activity. The left-lateral offsets of the main stream courses of the largest river basins, GPS measurements, and obliquity of plate convergence along the Cocos subduction zone in the Guerrero sector suggest the activity of sub-latitudinal left-lateral strike-slip faults. Notably, the regional left-lateral strike-slip fault that offsets the Papagayo River near the town of La Venta named "La Venta Fault" shows evidence of recent activity, corroborated also by GPS measurements (4-5 mm/year of sinistral motion). Assuming that during a probable earthquake the whole mapped length of this fault would rupture, it would produce an event of maximum moment magnitude Mw = 7.7. Even though only a few focal mechanism solutions indicate a stress regime relevant for reactivation of these strike-slip structures, we hypothesize that these faults are active and suggest two probable explanations: (1) these faults are characterized by long recurrence period, i.e., beyond the instrumental record, or (2) they experience slow slip events and/or associated fault creep. The analysis of focal mechanism solutions of small magnitude earthquakes in the upper plate, for the period between 1995

  6. Tetrandrine identified in a small molecule screen to activate mesenchymal stem cells for enhanced immunomodulation.

    Yang, Zijiang; Concannon, John; Ng, Kelvin S; Seyb, Kathleen; Mortensen, Luke J; Ranganath, Sudhir; Gu, Fangqi; Levy, Oren; Tong, Zhixiang; Martyn, Keir; Zhao, Weian; Lin, Charles P; Glicksman, Marcie A; Karp, Jeffrey M

    2016-07-26

    Pre-treatment or priming of mesenchymal stem cells (MSC) prior to transplantation can significantly augment the immunosuppressive effect of MSC-based therapies. In this study, we screened a library of 1402 FDA-approved bioactive compounds to prime MSC. We identified tetrandrine as a potential hit that activates the secretion of prostaglandin E2 (PGE2), a potent immunosuppressive agent, by MSC. Tetrandrine increased MSC PGE2 secretion through the NF-κB/COX-2 signaling pathway. When co-cultured with mouse macrophages (RAW264.7), tetrandrine-primed MSC attenuated the level of TNF-α secreted by RAW264.7. Furthermore, systemic transplantation of primed MSC into a mouse ear skin inflammation model significantly reduced the level of TNF-α in the inflamed ear, compared to unprimed cells. Screening of small molecules to pre-condition cells prior to transplantation represents a promising strategy to boost the therapeutic potential of cell therapy.

  7. Osteogenic Activity of Locally Applied Small Molecule Drugs in a Rat Femur Defect Model

    Jessica A. Cottrell

    2010-01-01

    Full Text Available The long-term success of arthroplastic joints is dependent on the stabilization of the implant within the skeletal site. Movement of the arthroplastic implant within the bone can stimulate osteolysis, and therefore methods which promote rigid fixation or bone growth are expected to enhance implant stability and the long-term success of joint arthroplasty. In the present study, we used a simple bilateral bone defect model to analyze the osteogenic activity of three small-molecule drug implants via microcomputerized tomography (micro-CT and histomorphometry. In this study, we show that local delivery of alendronate, but not lovastatin or omeprazole, led to significant new bone formation at the defect site. Since alendronate impedes osteoclast-development, it is theorized that alendronate treatment results in a net increase in bone formation by preventing osteoclast mediated remodeling of the newly formed bone and upregulating osteoblasts.

  8. Changes in lymphocyte subpopulations and adhesion/activation molecules following endotoxemia and major surgery

    Toft, P; Hokland, Marianne; Hansen, Tom Giedsing

    1995-01-01

    Major surgery as well as endotoxin-induced sepsis is accompanied by lymphocytopenia in peripheral blood. The purpose of this study was to investigate the redistribution of lymphocyte subpopulations and adhesion/activation molecules on lymphocytes. Twenty-four rats were included in the investigation....... Eight rats received an intraperitoneal injection of E. coli endotoxin (2 mg kg-1), eight rats had a sham operation performed while eight rats received isotonic saline and served as a control group. Blood samples were obtained by making an incision in the tail before and 2 and 5 h after surgery...... or administration of endotoxin or saline. After isolation of lymphocytes by gradient centrifugation, flow-cytometric immunophenotyping was performed using CD2, CD3, CD4, CD8, CD11/CD18, CD20, CD44 and MHC II monoclonal antibodies. Endotoxemia and surgery were both accompanied by increased serum cortisol...

  9. Benzimidazole-1,2,3-triazole hybrid molecules: synthesis and evaluation for antibacterial/antifungal activity.

    Ouahrouch, Abdelaaziz; Ighachane, Hana; Taourirte, Moha; Engels, Joachim W; Sedra, My Hassan; Lazrek, Hassan B

    2014-10-01

    A novel series of hybrid molecules 4a-i and 5a-i were prepared by condensation of 4-(trimethylsilylethynyl)benzaldehyde 1 with substituted o-phenylenediamines. These in turn were reacted with 2-(azidomethoxy)ethyl acetate in a Cu alkyne-azide cycloaddition (CuAAC) to generate the 1,2,3-triazole pharmacophore under microwave assistance. The newly synthesized compounds were examined for their in vitro antimicrobial activities against Gram-positive and Gram-negative bacteria and the phytopathogenic fungi Verticillium dahliae and Fusarium oxysporum f. sp. albedinis. 2-((4-(4-(5-Trifluoromethyl benzimidazol-2-yl)phenyl)-1,2,3-triazol-1-yl)methoxy)ethanol 5e showed a moderate inhibition of 30% in the Foa sporulation test.

  10. Microencapsulation effectiveness of small active molecules in biopolymer by ultrasonic atomization technique.

    Cascone, Sara; Lamberti, Gaetano; Titomanlio, Giuseppe; Barba, Anna Angela; d'Amore, Matteo

    2012-12-01

    A method to produce biopolymeric (alginate) microparticles by ultrasonic assisted atomization, previously developed, has been applied to the production of microparticles loaded with a small active molecule (theophylline). Fine loaded alginate droplets have been cross-linked with divalent ions to produce microparticles. Once produced, the particles have been separated by centrifugation or filtration and then they have been dried. Drug release has been evaluated by dissolution tests, dissolving the dried particles in acidic solution at pH 1 for a given time and then at pH 7 to simulate the stomach and intestinal environment, respectively. The encapsulation efficiency and the drug loading have been investigated and the operating conditions have been changed to clarify the role of the transport phenomena on the overall process. To increase the drug loading, shorter separation time and better network's structure were identified as the key operating parameters to allow the process to gain interest from a practical point of view.

  11. Auroral Electrojet Index Designed to Provide a Global Measure, l-minute Intervals, of Auroral Zone Magnetic Activity

    National Oceanic and Atmospheric Administration, Department of Commerce — The Auroral Electrojet index (AE) is designed to provide a global quantitative measure of auroral zone magnetic activity produced by enhanced ionospheric currents...

  12. Auroral Electrojet Indices Designed to Provide a Global Measure, 2.5-Minute Intervals, of Auroral Zone Magnetic Activity

    National Oceanic and Atmospheric Administration, Department of Commerce — The Auroral Electrojet index (AE) is designed to provide a global quantitative measure of auroral zone magnetic activity produced by enhanced ionospheric currents...

  13. Small molecule inhibition of 6-phosphofructo-2-kinase suppresses t cell activation

    Telang Sucheta

    2012-05-01

    Full Text Available Abstract Background T cell activation is associated with a rapid increase in intracellular fructose-2,6-bisphosphate (F2,6BP, an allosteric activator of the glycolytic enzyme, 6-phosphofructo-1-kinase. The steady state concentration of F2,6BP in T cells is dependent on the expression of the bifunctional 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatases (PFKFB1-4 and the fructose-2,6-bisphosphatase, TIGAR. Of the PFKFB family of enzymes, PFKFB3 has the highest kinase:bisphosphatase ratio and has been demonstrated to be required for T cell proliferation. A small molecule antagonist of PFKFB3, 3-(3-pyridinyl-1-(4-pyridinyl-2-propen-1-one (3PO, recently has been shown to reduce F2,6BP synthesis, glucose uptake and proliferation in transformed cells. We hypothesized that the induction of PFKFB3 expression may be required for the stimulation of glycolysis in T cells and that exposure to the PFKFB3 antagonist, 3PO, would suppress T cell activation. Methods We examined PFKFB1-4 and TIGAR expression and F2,6BP concentration in purified CD3+ T cells stimulated with microbead-conjugated agonist antibodies specific for CD3 and the co-stimulatory receptor, CD28. We then determined the effect of 3PO on anti-CD3/anti-CD28-induced T cell activation, F2,6BP synthesis, 2-[1-14C]-deoxy-d-glucose uptake, lactate secretion, TNF-α secretion and proliferation. Finally, we examined the effect of 3PO administration on the development of delayed type hypersensitivity to methylated BSA and on imiquimod-induced psoriasis in mice. Results We found that purified human CD3+ T cells express PFKFB2, PFKFB3, PFKFB4 and TIGAR, and that anti-CD3/anti-CD28 conjugated microbeads stimulated a >20-fold increase in F2,6BP with a coincident increase in protein expression of the PFKFB3 family member and a decrease in TIGAR protein expression. We then found that exposure to the PFKFB3 small molecule antagonist, 3PO (1–10 μM, markedly attenuated the stimulation of F2,6BP

  14. Crystallographic structure of a small molecule SIRT1 activator-enzyme complex

    Dai, Han; Case, April W.; Riera, Thomas V.; Considine, Thomas; Lee, Jessica E.; Hamuro, Yoshitomo; Zhao, Huizhen; Jiang, Yong; Sweitzer, Sharon M.; Pietrak, Beth; Schwartz, Benjamin; Blum, Charles A.; Disch, Jeremy S.; Caldwell, Richard; Szczepankiewicz, Bruce; Oalmann, Christopher; Yee Ng, Pui; White, Brian H.; Casaubon, Rebecca; Narayan, Radha; Koppetsch, Karsten; Bourbonais, Francis; Wu, Bo; Wang, Junfeng; Qian, Dongming; Jiang, Fan; Mao, Cheney; Wang, Minghui; Hu, Erding; Wu, Joe C.; Perni, Robert B.; Vlasuk, George P.; Ellis, James L.

    2015-07-01

    SIRT1, the founding member of the mammalian family of seven NAD+-dependent sirtuins, is composed of 747 amino acids forming a catalytic domain and extended N- and C-terminal regions. We report the design and characterization of an engineered human SIRT1 construct (mini-hSIRT1) containing the minimal structural elements required for lysine deacetylation and catalytic activation by small molecule sirtuin-activating compounds (STACs). Using this construct, we solved the crystal structure of a mini-hSIRT1-STAC complex, which revealed the STAC-binding site within the N-terminal domain of hSIRT1. Together with hydrogen-deuterium exchange mass spectrometry (HDX-MS) and site-directed mutagenesis using full-length hSIRT1, these data establish a specific STAC-binding site and identify key intermolecular interactions with hSIRT1. The determination of the interface governing the binding of STACs with human SIRT1 facilitates greater understanding of STAC activation of this enzyme, which holds significant promise as a therapeutic target for multiple human diseases.

  15. High pressure chemistry of red phosphorus by photo-activated simple molecules

    Ceppatelli, M.; Fanetti, S.; Bini, R.; Caporali, M.; Peruzzini, M.

    2014-05-01

    High pressure (HP) is very effective in reducing intermolecular distances and inducing unexpected chemical reactions. In addition the photo-activation of the reactants in HP conditions can lead to very efficient and selective processes. The chemistry of phosphorus is currently based on the white molecular form. The red polymeric allotrope, despite more stable and much less toxic, has not attracted much attention so far. However, switching from the white to the red form would benefit any industrial procedure, especially from an environmental point of view. On the other side, water and ethanol are renewable, environmental friendly and largely available molecules, usable as reactants and photo-activators in HP conditions. Here we report a study on the HP photo-induced reactivity of red phosphorus with water and ethanol, showing the possibility of very efficient and selective processes, leading to molecular hydrogen and valuable phosphorus compounds. The reactions have been studied by means of FTIR and Raman spectroscopy and pressure has been generated using membrane Diamond (DAC) and Sapphire (SAC) anvil cells. HP reactivity has been activated by the two-photon absorption of near-UV wavelengths and occurred in total absence of solvents, catalysts and radical initiators, at room T and mild pressure conditions (0.2-1.5 GPa).

  16. Studies of Active Ingredients in Cough Syrup by Capillary Zone Electrophoresis with Amperometric Detection

    ZHOU Tian-shu; WANG Ai-fang; WU Fang; SHI Guo-yue; FANG Yu-zhi

    2003-01-01

    The present paper covers a simple, reliable and reproducible method, based on capillary zone electrophoresis(CZE) with amperometric detection(AD), for the separation and the determination of ephedrine hydrochloride, promethazine hydrochloride and codeine phosphate. Under the optimal conditions, the three analytes were base-line separated completely within 16 min. Good linear relationships between the peak heights and the concentrations of the three analytes were obtained with the correlation coefficients better than 0.9993. The method was directly applied to the determination of the active ingredients in pharmaceutical preparations and the assay results were satisfactory.

  17. 77 FR 63320 - Agency Information Collection Activities: Petroleum Refineries in Foreign Trade Sub-zones

    2012-10-16

    ... in Foreign Trade Sub-zones AGENCY: U.S. Customs and Border Protection, Department of Homeland... Sub-zones. This is a proposed extension of an information collection that was previously approved. CBP... Trade Sub-zones. OMB Number: 1651-0063. Form Number: None. Abstract: The Foreign Trade Zones Act, 19...

  18. Impacts of stellar evolution and dynamics on the habitable zone: The role of rotation and magnetic activity

    Florian, Gallet; Louis, Amard; Sacha, Brun; Ana, Palacios; Stephane, Mathis

    2016-01-01

    In this article, we aim to provide the community with the dependence of the habitable zone upon the stellar mass, metallicity, rotation, and for various prescriptions of the limits of the habitable zone. We use the STAREVOL code to study the evolution of the habitable zone and of the continuously habitable zone limits. Mass and metallicity are the stellar parameters that have the most dramatic effects on the habitable zone limits. Conversely, for a given stellar mass and metallicity, stellar rotation has only a marginal effect on these limits and does not modify the width of the habitable zone. The evolution of the habitable zone limits is also correlated to the evolution of the stellar activity (through the Rossby number) that depends on the stellar mass considered. While the magnetic activity has negligible consequence in the case of more massive stars, these effects may have a strong impact on the habitability of a planet around M dwarf stars. Thus, stellar activity cannot be neglected and may have strong ...

  19. Small molecule regulation of self-association and catalytic activity in a supramolecular coordination complex.

    McGuirk, C Michael; Stern, Charlotte L; Mirkin, Chad A

    2014-03-26

    Herein, we report the synthesis and characterization of the first weak-link approach (WLA) supramolecular construct that employs the small molecule regulation of intermolecular hydrogen bonding interactions for the in situ control of catalytic activity. A biaryl urea group, prone to self-aggregation, was functionalized with a phosphinoalkyl thioether (P,S) hemilabile moiety and incorporated into a homoligated Pt(II) tweezer WLA complex. This urea-containing construct, which has been characterized by a single crystal X-ray diffraction study, can be switched in situ from a rigid fully closed state to a flexible semiopen state via Cl(-) induced changes in the coordination mode at the Pt(II) structural node. FT-IR and (1)H NMR spectroscopy studies were used to demonstrate that while extensive urea self-association persists in the flexible semiopen complex, these interactions are deterred in the rigid, fully closed complex because of geometric and steric restraints. Consequently, the urea moieties in the fully closed complex are able to catalyze a Diels-Alder reaction between cyclopentadiene and methyl vinyl ketone to generate 2-acetyl-5-norbornene. The free urea ligand and the semiopen complex show no such activity. The successful incorporation and regulation of a hydrogen bond donating catalyst in a WLA construct open the doors to a vast and rapidly growing catalogue of allosteric catalysts for applications in the detection and amplification of organic analytes.

  20. Discovery of Novel Small Molecules that Activate Satellite Cell Proliferation and Enhance Repair of Damaged Muscle.

    Billin, Andrew N; Bantscheff, Marcus; Drewes, Gerard; Ghidelli-Disse, Sonja; Holt, Jason A; Kramer, Henning F; McDougal, Alan J; Smalley, Terry L; Wells, Carrow I; Zuercher, William J; Henke, Brad R

    2016-02-19

    Skeletal muscle progenitor stem cells (referred to as satellite cells) represent the primary pool of stem cells in adult skeletal muscle responsible for the generation of new skeletal muscle in response to injury. Satellite cells derived from aged muscle display a significant reduction in regenerative capacity to form functional muscle. This decrease in functional recovery has been attributed to a decrease in proliferative capacity of satellite cells. Hence, agents that enhance the proliferative abilities of satellite cells may hold promise as therapies for a variety of pathological settings, including repair of injured muscle and age- or disease-associated muscle wasting. Through phenotypic screening of isolated murine satellite cells, we identified a series of 2,4-diaminopyrimidines (e.g., 2) that increased satellite cell proliferation. Importantly, compound 2 was effective in accelerating repair of damaged skeletal muscle in an in vivo mouse model of skeletal muscle injury. While these compounds were originally prepared as c-Jun N-terminal kinase 1 (JNK-1) inhibitors, structure-activity analyses indicated JNK-1 inhibition does not correlate with satellite cell activity. Screening against a broad panel of kinases did not result in identification of an obvious molecular target, so we conducted cell-based proteomics experiments in an attempt to identify the molecular target(s) responsible for the potentiation of the satellite cell proliferation. These data provide the foundation for future efforts to design improved small molecules as potential therapeutics for muscle repair and regeneration.

  1. Antibacterial activity and characteristics of modified ferrite powder coated with a gemini pyridinium salt molecule.

    Shirai, Akihiro; Maeda, Takuya; Ohkita, Motoaki; Nagamune, Hideaki; Kourai, Hiroki

    2007-09-01

    This report describes the synthesis of an antibacterial material consisting of a gemini quaternary ammonium salt (gemini-QUAT) immobilized on ferrite powder, and its antibacterial activity. A gemini-QUAT containing two pyridinium residues per molecule, 4,4'-[1,3-(2,2-dihydroxylmethyl-1,3-dithiapropane)]bis (1-octylpyridinium bromide), was immobilized on ferrite powder by a reaction between the hydroxyl group of the QUAT and trimethoxysilane. Immobilization of the gemini-QUAT on ferrite (F-gemini-QUAT) was confirmed when the dye, bromophenol blue, was released from F-gemini-QUAT-dye after contact between ferrite and the dye. Elemental analysis of the QUAT-ferrite determined the molar amount of QUAT on the ferrite. The antibacterial effect of the ferrite was investigated using a batch treatment system, and this effect was compared with that of another QUAT-ferrite (F-mono-QUAT) binding a mono-QUAT, which possesses one pyridinium residue, prepared by the same immobilization method as F-gemini-QUAT. Results indicated the F-gemini QUAT possessed a higher bactericidal potency and broader antibacterial spectrum compared to F-mono-QUAT. In addition, this study suggested that gemini-QUATs possessed high bactericidal potency without being influenced by immobilization to materials, and the antibacterial activity and characteristics of F-gemini-QUAT could be attributed to the unique structure of the immobilized gemini-QUAT.

  2. Fasciola hepatica Kunitz type molecule decreases dendritic cell activation and their ability to induce inflammatory responses.

    Cristian R Falcón

    Full Text Available The complete repertoire of proteins with immunomodulatory activity in Fasciola hepatica (Fh has not yet been fully described. Here, we demonstrated that Fh total extract (TE reduced LPS-induced DC maturation, and the DC ability to induce allogeneic responses. After TE fractionating, a fraction lower than 10 kDa (F<10 kDa was able to maintain the TE properties to modulate the DC pro- and anti-inflammatory cytokine production induced by LPS. In addition, TE or F<10 kDa treatment decreased the ability of immature DC to stimulate the allogeneic responses and induced a novo allogeneic CD4+CD25+Foxp3+ T cells. In contrast, treatment of DC with T/L or F<10 kDa plus LPS (F<10/L induced a regulatory IL-27 dependent mechanism that diminished the proliferative and Th1 and Th17 allogeneic responses. Finally, we showed that a Kunitz type molecule (Fh-KTM, present in F<10 kDa, was responsible for suppressing pro-inflammatory cytokine production in LPS-activated DC, by printing tolerogenic features on DC that impaired their ability to induce inflammatory responses. These results suggest a modulatory role for this protein, which may be involved in the immune evasion mechanisms of the parasite.

  3. Study on Integrated Recurrence Behaviors of Strong Earthquakes Along Entire Active Fault Zones in the Sichuan-Yunnan Region, China

    Yi Guixi; Wen Xueze; Xu Xiwei

    2003-01-01

    Based on historical earthquake data, we use statistical methods to study integrated recurrencebehaviors of strong earthquakes along 7 selected active fault zones in the Sichuan-Yunnanregion. The results show that recurrences of strong earthquakes in the 7 fault zones displaynear-random, random and clustering behaviors. The recurrence processes are never quasi-periodic, and are neither strength-time nor time-strength dependent. The more independentsegments for strong earthquake rupturing a fault zone has, the more complicated thecorresponding recurrence process is. And relatively active periods and quiescent periods forearthquake activity occur alternatively. Within the active periods, the distribution ofrecurrence time intervals between earthquakes has relatively large discretion, and can bemodelled well by a Weibull distribution. The time distribution of the quiescent periods hasrelatively small discretion, and can be approximately described by some distributions as thenormal. Both the durations of the active periods and the numbers of strong earthquakes withinthe active periods vary obviously cycle by cycle, leading to the relatively active periods havingnever repeated quasi-periodically. Therefore, the prohabilistic assessment for middle- and long-term seismic hazard for entireties of active fault zones based on data of historical strongearthquakes on the fault zones still faces difficulty.

  4. Characteristics of Relocated Quiet Zones Using Virtual Microphone Algorithm in an Active Headrest System

    Seokhoon Ryu

    2016-01-01

    Full Text Available This study displays theoretical and experimental investigation on the characteristics of the relocated zone of quiet by a virtual microphone (VM based filtered-x LMS (FxLMS algorithm which can be embedded in a real-time digital controller for an active headrest system. The attenuation changes at the relocated zones of quiet by the variation of the distance between the ear and the error microphone are mainly examined. An active headrest system was implemented for the control experiment at a chair and consists of two (left and right secondary loudspeakers, two error microphones, two observer microphones at ear positions in a HATS, and other electronics including a dSPACE 1401 controller. The VM based FxLMS algorithm achieved an attenuation of about 22 dB in the control experiment against a narrowband primary noise by the variation of the distance between the ear and the error microphone. The important factors for the algorithm are discussed as well.

  5. Earthworm activities in cassava and egusi melon fields in the transitional zone of Benin: linking farmers' perceptions with field studies

    Saïdou, A.; Kossou, D.; Brussaard, L.; Richards, P.; Kuyper, T.W.

    2008-01-01

    Farmers' perceptions of earthworm activities were studied in the transitional zone of Benin and linked to scientific explanations of earthworm casting activities. Earthworm activity was assessed in farmers' fields with three different cassava cultivars and in a field experiment with three different

  6. L1 cell adhesion molecule induces melanoma cell motility by activation of mitogen-activated protein kinase pathways.

    Yi, Young-Su; Baek, Kwang-Soo; Cho, Jae Youl

    2014-06-01

    L1 cell adhesion molecule (L1CAM) is highly expressed in various types of cancer cells and has been implicated in the control of cell proliferation and motility. Recently, L1CAM was reported to induce the motility of melanoma cells, but the mechanism of this induction remains poorly understood. In this study, we investigated the molecular mechanisms by which L1CAM induces the motility of melanoma cells. Unlike other types of cancer cells, B16F10 melanoma cells highly expressed L1CAM at both the RNA and protein levels, and the expression of L1CAM induced AP-1 activity. In accordance to AP-1 activation, MAPK signaling pathways were activated by L1CAM. Inhibition of L1CAM expression by L1CAM-specific siRNA suppressed the activation of MAPKs such as ERK and p38. However, no significant change was observed in JNK activation. As expected, upstream MAP2K, MKK3/6, MAP3K, and TAK1 were also deactivated by the inhibition of L1CAM expression. L1CAM induced the motility of B16F10 cells. Inhibition of L1CAM expression suppressed migration and invasion of B16F10 cells, but no suppressive effect was observed on their proliferation and anti-apoptotic resistance. Treatment of B16F10 cells with U0126, an ERK inhibitor, or SB203580, a p38 inhibitor, suppressed the migration and invasion abilities of B16F10 cells. Taken together, our results suggest that L1CAM induces the motility of B16F10 melanoma cells via the activation of MAPK pathways. This finding provides a more detailed molecular mechanism of L1CAM-mediated induction of melanoma cell motility.

  7. Antimicrobial activity of innate immune molecules against Streptococcus pneumoniae, Moraxella catarrhalis and nontypeable Haemophilus influenzae

    Teufert Karen

    2004-05-01

    Full Text Available Abstract Background Despite its direct connection to the nasopharynx which harbors otitis media pathogens as part of its normal flora, the middle ear cavity is kept free of these bacteria by as yet unknown mechanisms. Respiratory mucosal epithelia, including those of the middle ear and eustachian tube, secrete antimicrobial effectors including lysozyme, lactoferrin and β defensins-1 and -2. To elucidate the role of these innate immune molecules in the normal defense and maintenance of sterility of respiratory mucosa such as that of the middle ear, we assessed their effect on the respiratory pathogens nontypeable Haemophilus influenzae (NTHi 12, Moraxella catarrhalis 035E, and Streptococcus pneumoniae 3, and 6B. Methods Two assay methods, the radial assay and the liquid broth assay, were employed for testing the antimicrobial activity of the molecules. This was done in order to minimize the possibility that the observed effects were artifacts of any single assay system employed. Also, transmission electron microscopy (TEM was employed to evaluate the effect of antimicrobial innate immune molecules on OM pathogens. For the statistical analysis of the data, Student's t-test was performed. Results Results of the radial diffusion assay showed that β defensin-2 was active against all four OM pathogens tested, while treatment with β defensin-1 appeared to only affect M. catarrhalis. The radial assay results also showed that lysozyme was quite effective against S. pneumoniae 3 and 6B and was partially bacteriostatic/bactericidal against M. catarrhalis. Lysozyme however, appeared not to affect the growth of NTHi. Thus, lysozyme seems to have a more pronounced impact on the growth of the Gram-positive S. pneumoniae as compared to that of Gram-negative pathogens. Lactoferrin on the other hand, enhanced the growth of the bacteria tested. The results of the radial assays were confirmed using liquid broth assays for antimicrobial activity, and showed that

  8. How Large Scale Flows in the Solar Convection Zone may Influence Solar Activity

    Hathaway, D. H.

    2004-01-01

    Large scale flows within the solar convection zone are the primary drivers of the Sun s magnetic activity cycle. Differential rotation can amplify the magnetic field and convert poloidal fields into toroidal fields. Poleward meridional flow near the surface can carry magnetic flux that reverses the magnetic poles and can convert toroidal fields into poloidal fields. The deeper, equatorward meridional flow can carry magnetic flux toward the equator where it can reconnect with oppositely directed fields in the other hemisphere. These axisymmetric flows are themselves driven by large scale convective motions. The effects of the Sun s rotation on convection produce velocity correlations that can maintain the differential rotation and meridional circulation. These convective motions can influence solar activity themselves by shaping the large-scale magnetic field pattern. While considerable theoretical advances have been made toward understanding these large scale flows, outstanding problems in matching theory to observations still remain.

  9. Direct Measurements of Local Coupling between Myosin Molecules Are Consistent with a Model of Muscle Activation.

    Sam Walcott

    2015-11-01

    Full Text Available Muscle contracts due to ATP-dependent interactions of myosin motors with thin filaments composed of the proteins actin, troponin, and tropomyosin. Contraction is initiated when calcium binds to troponin, which changes conformation and displaces tropomyosin, a filamentous protein that wraps around the actin filament, thereby exposing myosin binding sites on actin. Myosin motors interact with each other indirectly via tropomyosin, since myosin binding to actin locally displaces tropomyosin and thereby facilitates binding of nearby myosin. Defining and modeling this local coupling between myosin motors is an open problem in muscle modeling and, more broadly, a requirement to understanding the connection between muscle contraction at the molecular and macro scale. It is challenging to directly observe this coupling, and such measurements have only recently been made. Analysis of these data suggests that two myosin heads are required to activate the thin filament. This result contrasts with a theoretical model, which reproduces several indirect measurements of coupling between myosin, that assumes a single myosin head can activate the thin filament. To understand this apparent discrepancy, we incorporated the model into stochastic simulations of the experiments, which generated simulated data that were then analyzed identically to the experimental measurements. By varying a single parameter, good agreement between simulation and experiment was established. The conclusion that two myosin molecules are required to activate the thin filament arises from an assumption, made during data analysis, that the intensity of the fluorescent tags attached to myosin varies depending on experimental condition. We provide an alternative explanation that reconciles theory and experiment without assuming that the intensity of the fluorescent tags varies.

  10. Antiviral activity of a small molecule deubiquitinase inhibitor occurs via induction of the unfolded protein response.

    Jeffrey W Perry

    Full Text Available Ubiquitin (Ub is a vital regulatory component in various cellular processes, including cellular responses to viral infection. As obligate intracellular pathogens, viruses have the capacity to manipulate the ubiquitin (Ub cycle to their advantage by encoding Ub-modifying proteins including deubiquitinases (DUBs. However, how cellular DUBs modulate specific viral infections, such as norovirus, is poorly understood. To examine the role of DUBs during norovirus infection, we used WP1130, a small molecule inhibitor of a subset of cellular DUBs. Replication of murine norovirus in murine macrophages and the human norovirus Norwalk virus in a replicon system were significantly inhibited by WP1130. Chemical proteomics identified the cellular DUB USP14 as a target of WP1130 in murine macrophages, and pharmacologic inhibition or siRNA-mediated knockdown of USP14 inhibited murine norovirus infection. USP14 is a proteasome-associated DUB that also binds to inositol-requiring enzyme 1 (IRE1, a critical mediator of the unfolded protein response (UPR. WP1130 treatment of murine macrophages did not alter proteasome activity but activated the X-box binding protein-1 (XBP-1 through an IRE1-dependent mechanism. In addition, WP1130 treatment or induction of the UPR also reduced infection of other RNA viruses including encephalomyocarditis virus, Sindbis virus, and La Crosse virus but not vesicular stomatitis virus. Pharmacologic inhibition of the IRE1 endonuclease activity partially rescued the antiviral effect of WP1130. Taken together, our studies support a model whereby induction of the UPR through cellular DUB inhibition blocks specific viral infections, and suggest that cellular DUBs and the UPR represent novel targets for future development of broad spectrum antiviral therapies.

  11. Active faults in the deformation zone off Noto Peninsula, Japan, revealed by high- resolution seismic profiles

    Inoue, T.; Okamura, Y.; Murakami, F.; Kimura, H.; Ikehara, K.

    2008-12-01

    Recently, a lot of earthquakes occur in Japan. The deformation zone which many faults and folds have concentrated exists on the Japan Sea side of Japan. The 2007 Noto Hanto Earthquake (MJMA 6.9) and 2007 Chuetsu-oki Earthquake (MJMA 6.8) were caused by activity of parts of faults in this deformation zone. The Noto Hanto Earthquake occurred on 25 March, 2007 under the northwestern coast of Noto Peninsula, Ishikawa Prefecture, Japan. This earthquake is located in Quaternary deformation zone that is continued from northern margin of Noto Peninsula to southeast direction (Okamura, 2007a). National Institute of Advanced Industrial Science and Technology (AIST) carried out high-resolution seismic survey using Boomer and 12 channels short streamer cable in the northern part off Noto Peninsula, in order to clarify distribution and activities of active faults in the deformation zone. A twelve channels short streamer cable with 2.5 meter channel spacing developed by AIST and private corporation is designed to get high resolution seismic profiles in shallow sea area. The multi-channel system is possible to equip on a small fishing boat, because the data acquisition system is based on PC and the length of the cable is short and easy to handle. Moreover, because the channel spacing is short, this cable is very effective for a high- resolution seismic profiling survey in the shallow sea, and seismic data obtained by multi-channel cable can be improved by velocity analysis and CDP stack. In the northern part off Noto Peninsula, seismic profiles depicting geologic structure up to 100 meters deep under sea floor were obtained. The most remarkable reflection surface recognized in the seismic profiles is erosion surface at the Last Glacial Maximum (LGM). In the western part, sediments about 30 meters (40 msec) thick cover the erosional surface that is distributed under the shelf shallower than 100m in depth and the sediments thin toward offshore and east. Flexures like deformation in

  12. TLR-induced activation of Btk- Role for endosomal MHC class Ⅱ molecules revealed

    Joan Ni Gabhann; Caroline A Jefferies

    2011-01-01

    @@ MHC molecules have been shown to play key roles in the immune system including regulating T-cell repertoire development through the process of positive and negative selection.MHC molecules also function to bridge the innate and adaptive immune system through the presentation of processed antigenic peptides to T-cells.

  13. NMDA modulates oligodendrocyte differentiation of subventricular zone cells through PKC activation

    Fabio eCavaliere

    2013-12-01

    Full Text Available Multipotent cells from the juvenile subventricular zone (SVZ possess the ability to differentiate into new neural cells. Depending on local signals, SVZ can generate new neurons, astrocytes or oligodendrocytes. We previously demonstrated that activation of NMDA receptors in SVZ progenitors increases the rate of oligodendrocyte differentiation. Here we investigated the mechanisms involved in NMDA receptor-dependent differentiation. Using functional studies performed with the reporter gene luciferase we found that activation of NMDA receptor stimulates PKC. In turn, stimulation of PKC precedes the activation of NADPH oxidase (NOX as demonstrated by translocation of the p67phox subunit to the cellular membrane. We propose that NOX2 is involved in the transduction of the signal from NMDA receptors through PKC activation as the inhibitor gp91 reduced their pro-differentiation effect. In addition, our data and that from other groups suggest that signaling through the NMDA receptor/PKC/NOX2 cascade generates ROS that activate the PI3/mTOR pathway and finally leads to the generation of new oligodendrocytes.

  14. Hedgehog Controls Quiescence and Activation of Neural Stem Cells in the Adult Ventricular-Subventricular Zone

    Mathieu Daynac

    2016-10-01

    Full Text Available Identifying the mechanisms controlling quiescence and activation of neural stem cells (NSCs is crucial for understanding brain repair. Here, we demonstrate that Hedgehog (Hh signaling actively regulates different pools of quiescent and proliferative NSCs in the adult ventricular-subventricular zone (V-SVZ, one of the main brain neurogenic niches. Specific deletion of the Hh receptor Patched in NSCs during adulthood upregulated Hh signaling in quiescent NSCs, progressively leading to a large accumulation of these cells in the V-SVZ. The pool of non-neurogenic astrocytes was not modified, whereas the activated NSC pool increased after a short period, before progressively becoming exhausted. We also showed that Sonic Hedgehog regulates proliferation of activated NSCs in vivo and shortens both their G1 and S-G2/M phases in culture. These data demonstrate that Hh orchestrates the balance between quiescent and activated NSCs, with important implications for understanding adult neurogenesis under normal homeostatic conditions or during injury.

  15. Location of high seismic activity zones and seismic hazard assessment in Zabrze Bielszowice coal mine using passive tomography

    LURKA A

    2008-01-01

    In the paper results of passive tomography calculations have been presented to assess rockburst hazard and locate high seismic activity zones in the vicinity of longwall 306 in Zabrze Bielszowice coal mine. The area of study was 1000 m in X direction by 900 m in Y direction. The zones of high values of P-wave propagation velocity have been found to correlate with the distribution of large seismic tremors.

  16. The new generation drug candidate molecules: Spectral, electrochemical, DNA-binding and anticancer activity properties

    Gölcü, Ayşegül; Muslu, Harun; Kılıçaslan, Derya; Çeşme, Mustafa; Eren, Özge; Ataş, Fatma; Demirtaş, İbrahim

    2016-09-01

    The new generation drug candidate molecules [Cu(5-Fu)2Cl2H2O] (NGDCM1) and [Zn(5-Fu)2(CH3COO)2] (NGDCM2) were obtained from the reaction of copper(II) and zinc(II) salts with the anticancer drug 5-fluoracil (5-Fu). These compounds have been characterized by spectroscopic and analytical techniques. Thermal behavior of the compounds were also investigated. The electrochemical properties of the compounds have been investigated by cyclic voltammetry (CV) using glassy carbon electrode. The biological activity of the NGDCM1 and NGDCM2 has been evaluated by examining their ability to bind to fish sperm double strand DNA (FSdsDNA) with UV spectroscopy. UV studies of the interaction of the 5-Fu and metal derivatives with FSdsDNA have shown that these compounds can bind to FSdsDNA. The binding constants of the compounds with FSdsDNA have also been calculated. Thermal decomposition of the compounds lead to the formation of CuO and ZnO as final products. The effect of proliferation 5-Fu, NGDCM1 and NGDCM2 were examined on the HeLa cells using real-time cell analyzer with three different concentrations.

  17. Small-molecule inhibition of MLL activity by disruption of its interaction with WDR5.

    Senisterra, Guillermo; Wu, Hong; Allali-Hassani, Abdellah; Wasney, Gregory A; Barsyte-Lovejoy, Dalia; Dombrovski, Ludmila; Dong, Aiping; Nguyen, Kong T; Smil, David; Bolshan, Yuri; Hajian, Taraneh; He, Hao; Seitova, Alma; Chau, Irene; Li, Fengling; Poda, Gennadiy; Couture, Jean-François; Brown, Peter J; Al-Awar, Rima; Schapira, Matthieu; Arrowsmith, Cheryl H; Vedadi, Masoud

    2013-01-01

    WDR5 (WD40 repeat protein 5) is an essential component of the human trithorax-like family of SET1 [Su(var)3-9 enhancer-of-zeste trithorax 1] methyltransferase complexes that carry out trimethylation of histone 3 Lys4 (H3K4me3), play key roles in development and are abnormally expressed in many cancers. In the present study, we show that the interaction between WDR5 and peptides from the catalytic domain of MLL (mixed-lineage leukaemia protein) (KMT2) can be antagonized with a small molecule. Structural and biophysical analysis show that this antagonist binds in the WDR5 peptide-binding pocket with a Kd of 450 nM and inhibits the catalytic activity of the MLL core complex in vitro. The degree of inhibition was enhanced at lower protein concentrations consistent with a role for WDR5 in directly stabilizing the MLL multiprotein complex. Our data demonstrate inhibition of an important protein-protein interaction and form the basis for further development of inhibitors of WDR5-dependent enzymes implicated in MLL-rearranged leukaemias or other cancers.

  18. NK cells, displaying early activation, cytotoxicity and adhesion molecules, are associated with mild dengue disease.

    Azeredo, E L; De Oliveira-Pinto, L M; Zagne, S M; Cerqueira, D I S; Nogueira, R M R; Kubelka, C F

    2006-02-01

    During the innate immune response against infections, Natural Killer (NK) cells are as important effector cells as are Cytotoxic T lymphocytes (CTL) generated after antigenic stimulation in the adaptative response. NK cells increase in numbers, after viral infection or vaccination. We investigated the NK cell and CD8 T lymphocyte status in 55 dengue infected patients. The NK (CD56+CD3-) and CD56+ T cell (CD56+CD3+) rates rise during the acute phase of disease. The majority of NK cells from dengue patients display early markers for activation (CD69, HLA-DR, and CD38) and cell adhesion molecules (CD44, CD11a) during the acute phase of disease. The intracellular cytotoxic granule, TIA-1, is also up-regulated early in NK cells. Most of these markers appear also on CD8+ T lymphocytes but during the late acute phase. Circulating IL-15 is elevated in a significant number of patients during early acute infection and its values were statistically correlated with NK frequencies and cytotoxic markers on NKs. We have therefore shown that dengue virus infection is very likely stimulating a cytotoxic response that may be efficient in controlling the virus in synergism with CD8+ T lymphocytes. Interestingly, the heightened CD56+CD3-, CD56+CD3+, CD56+TIA-1+ and CD56+CD11a+ cell rates are associated with mild dengue clinical manifestations and might indicate a good prognosis of the disease.

  19. NK cells, displaying early activation, cytotoxicity and adhesion molecules, are associated with mild dengue disease

    Azeredo, E L; De Oliveira-Pinto, L M; Zagne, S M; Cerqueira, D I S; Nogueira, R M R; Kubelka, C F

    2006-01-01

    During the innate immune response against infections, Natural Killer (NK) cells are as important effector cells as are Cytotoxic T lymphocytes (CTL) generated after antigenic stimulation in the adaptative response. NK cells increase in numbers, after viral infection or vaccination. We investigated the NK cell and CD8 T lymphocyte status in 55 dengue infected patients. The NK (CD56+CD3−) and CD56+ T cell (CD56+CD3+) rates rise during the acute phase of disease. The majority of NK cells from dengue patients display early markers for activation (CD69, HLA-DR, and CD38) and cell adhesion molecules (CD44, CD11a) during the acute phase of disease. The intracellular cytotoxic granule, TIA-1, is also up-regulated early in NK cells. Most of these markers appear also on CD8+ T lymphocytes but during the late acute phase. Circulating IL-15 is elevated in a significant number of patients during early acute infection and its values were statistically correlated with NK frequencies and cytotoxic markers on NKs. We have therefore shown that dengue virus infection is very likely stimulating a cytotoxic response that may be efficient in controlling the virus in synergism with CD8+ T lymphocytes. Interestingly, the heightened CD56+CD3−, CD56+CD3+, CD56+TIA-1+ and CD56+CD11a+ cell rates are associated with mild dengue clinical manifestations and might indicate a good prognosis of the disease. PMID:16412060

  20. Kinetics of T cell-activation molecules in response to Mycobacterium tuberculosis antigens

    Antas Paulo RZ

    2002-01-01

    Full Text Available The phenotypic features acquired subsequent to antigen-specific stimulation in vitro were evaluated by means of the kinetic expressions of CD69 and CD25 activation molecules on T lymphocytes and assayed by flow cytometry in response to PPD, Ag85B, and ferritin in PPD-positive healthy control individuals. In response to PHA, CD69 staining on both CD4+ and CD8+ T cells became initially marked after 4 h, peaked at 24 h, and quickly decreased after 120 h. For CD25, a latter expression was detected around 8 h, having increased after 96 h. As expected, the response rate to the mycobacterial antigens was much lower than that to the mitogen. Positive staining was high after 96 h for CD25 and after 24 h for CD69. CD69 expression was significantly enhanced (p < 0.05 on CD8+ as compared to CD4+ T cells. High levels were also found between 96-120 h. Regarding Ag85B, CD25+ cells were mostly CD4+ instead of CD8+ T cells. Moreover, in response to ferritin, a lower CD25 expression was noted. The present data will allow further characterization of the immune response to new mycobacterial-specific antigens and their evaluation for possible inclusion in developing new diagnostic techniques for tuberculosis as well in a new vaccine to prevent the disease.

  1. Tailoring activated carbon by surface chemical modification with O, S, and N containing molecules

    Rachel RibeiroVieira Azzi Rios

    2003-06-01

    Full Text Available In this work the surface of activated carbon was chemically modified in order to introduce O, S and N containing groups. The activated carbon surface was selectively oxidized with concentrated HNO3 under controlled conditions. Characterization by thermogravimetric analyses, infrared spectroscopy and NaOH titration suggested the formation of mainly -COOH and small amounts of -OH groups, with concentration of approximately 4.10(21 groups/g of carbon. These -COOH functionalized carbons showed high adsorption capacity for metal cations in aqueous solution in the following order: Pb+2>Cu+2>Ni+2 >Cd+2~Co+2>Ca+2 , suggesting a cation exchange mechanism via a surface complex [COO-M+2]. These -COOHsurf groups can be reacted with SOCl2 to produce a surface acylchloride group, -COCl. This surface -COCl group proved to be a very reactive and versatile intermediate for the grafting of different S and N containing molecules onto the carbon surface, such as 1,2-ethaneditiol (EDT-, HSCH2CH2SH 1,7-dimercapto-4-thioheptane (DMTH-HSCH2CH2CH2SCH2CH 2CH2SH or 1,2-ethylenediamine (EDA- NH2CH2CH2NH2 and triethyltetraamine, TEA (H2NCH2CH2NHCH2CH 2NHCH2CH2 NH2. The characterization of these materials was carried out by TG, IR and TPDMS (Temperature Programmed Decomposition Mass Spectrometry experiments suggesting the formation of thioesther and amide surface groups, i.e. -COSR and -CONHR, with yields of approximately 50 and 75% for the reaction with DME and EDA, respectively. Preliminary adsorption experiments showed that these materials can efficiently remove metals such as Pb+2, Cu+2 and Ni+2 from aqueous medium.

  2. 77 FR 47429 - Agency Information Collection Activities; Petroleum Refineries in Foreign Trade Sub-zones

    2012-08-08

    ... in Foreign Trade Sub-zones AGENCY: U.S. Customs and Border Protection (CBP), Department of Homeland... concerning the Petroleum Refineries in Foreign Trade Sub-zones. This request for comment is being made... Refineries in Foreign Trade Sub-zones. OMB Number: 1651-0063. Form Number: None. Abstract: The Foreign...

  3. Pedestrian-oriented zoning is associated with reduced income and poverty disparities in adult active travel to work, United States.

    Chriqui, Jamie F; Leider, Julien; Thrun, Emily; Nicholson, Lisa M; Slater, Sandy J

    2017-02-01

    Active travel to work can provide additional minutes of daily physical activity. While the literature points to the relationship between zoning, equity and socioeconomic status, and physical activity, no study has quantitatively explored these connections. This study examined whether zoning may help to moderate any income and poverty inequities in active travel and taking public transit to work. Research was conducted between May 2012 and June 2015. Zoning data were compiled for 3914 jurisdictions covering 45.45% of the U.S. population located in 471 of the most populous U.S. counties and 2 consolidated cities located in 48 states and the District of Columbia. (Sensitivity analyses also captured unincorporated areas which, with the municipalities, collectively covered ~72% of the U.S.

  4. Complement activating soluble pattern recognition molecules with collagen-like regions, mannan-binding lectin, ficolins and associated proteins

    Thiel, Steffen

    2007-01-01

    Mannan-binding lectin (MBL), L-ficolin, M-ficolin and H-ficolin are all complement activating soluble pattern recognition molecules with recognition domains linked to collagen-like regions. All four may form complexes with four structurally related proteins, the three MBL-associated serine...... proteases (MASPs), MASP-1, MASP-2 and MASP-3, and a smaller MBL-associated protein (MAp19). The four recognition molecules recognize patterns of carbohydrate or acetyl-group containing ligands. After binding to the relevant targets all four are able to activate the complement system. We thus have a system...... where four different and/or overlapping patterns of microbial origin or patterns of altered-self may be recognized, but in all cases the signalling molecules, the MASPs, are shared. MASP-1 and MASP-3 are formed from one gene, MASP1/3, by alternative splicing generating two different mRNAs from a single...

  5. EEG reactions of the human brain in the gradient magnetic field zone of the active geological fault (pilot study)

    Pobachenko, S. V.; Shitov, A. V.; Grigorjev, P. E.; Sokolov, M. V.; Zubrilkin, A. I.; Vypiraylo, D. N.; Solovjev, A. V.

    2016-12-01

    This paper presents the results of experimental studies of the dynamics of the functional state of a person within the zone of an active geological fault characterized by abnormal spatial distribution of the magnetic- field vector values. It is shown that these geophysical modifications have a pronounced effect on the fluctuations of the electrical activity of the human brain. When the person gets into a zone with abnormal levels of gradient magnetic field in the absence of any subjective sensations, a nonspecific orientation activation reaction is observed, which is characterized by a significant increase in the levels of peak performance in key functional EEG frequency bands.

  6. SAD-B Phosphorylation of CAST Controls Active Zone Vesicle Recycling for Synaptic Depression

    Sumiko Mochida

    2016-09-01

    Full Text Available Short-term synaptic depression (STD is a common form of activity-dependent plasticity observed widely in the nervous system. Few molecular pathways that control STD have been described, but the active zone (AZ release apparatus provides a possible link between neuronal activity and plasticity. Here, we show that an AZ cytomatrix protein CAST and an AZ-associated protein kinase SAD-B coordinately regulate STD by controlling reloading of the AZ with release-ready synaptic vesicles. SAD-B phosphorylates the N-terminal serine (S45 of CAST, and S45 phosphorylation increases with higher firing rate. A phosphomimetic CAST (S45D mimics CAST deletion, which enhances STD by delaying reloading of the readily releasable pool (RRP, resulting in a pool size decrease. A phosphonegative CAST (S45A inhibits STD and accelerates RRP reloading. Our results suggest that the CAST/SAD-B reaction serves as a brake on synaptic transmission by temporal calibration of activity and synaptic depression via RRP size regulation.

  7. Diabatic heating profiles over the continental convergence zone during the monsoon active spells

    Chattopadhyay, Rajib; Sur, Sharmila; Joseph, Susmitha; Sahai, A. K.

    2013-07-01

    The present paper aims to bring out the robust common aspects of spatio-temporal evolution of diabatic heating during the monsoon intraseasonal active phases over the continental tropical convergence zone (CTCZ). The robustness of spatio-temporal features is determined by comparing the two state-of-the art reanalyses: NCEP Climate Forecast System reanalysis and Modern ERA Retrospective Analysis. The inter-comparison is based on a study period of 26 years (1984-2009). The study confirms the development of deep heating over the CTCZ region during the active phase and is consistent between the two datasets. However, the detailed temporal evolution of the vertical structure (e.g., vertical tilts) of heating differs at times. The most important common feature from both the datasets is the significant vertical redistribution of heating with the development of shallow (low level) heating and circulation over the CTCZ region 3-7 days after the peak active phase. The shallow circulation is found to be associated with increased vertical shear and relative vorticity over certain regions in the subcontinent. This increased vertical shear and relative vorticity in the lower levels could be crucial in the sustenance of rainfall after the peak active phase. Model experiments with linear dynamics affirm the role of shallow convection in increasing the lower level circulation as observed.

  8. Hydrothermal quartz formation during fluctuations of brittle shear-zone activity and fluid flow: grain growth and deformation structures of the Pfahl shear zone (Germany)

    Yilmaz, T.; Prosser, G.; Liotta, D.; Kruhl, J. H.

    2012-12-01

    The Bavarian Pfahl shear zone is a WNW-ESE trending dextral strike-slip shear zone at the SW margin of the Bohemian Massif (Central Europe). It was discontinuously active during decreasing PT-conditions, i.e. from ductile to brittle, from the late-Carboniferous to the late-Cretaceous - Paleocene times. Triassic hydrothermal activity produced a 150 km long and 30-100 m wide quartz dyke along the main fault, surrounded by sheared basement rocks. Within a zone of >10 m metasomatism transformed the wall rocks to mostly kaolinite, chlorite and phyllosilicates. The quartz dyke exhibits a layered to lenticular and partly symmetric structure with different types of quartz masses, transected by a complex quartz vein network. This already indicates pulses of fluid flux and fragmentation during the lifetime of the shear zone. Analyses by optical microscopy, cathodoluminescence (CL) and SEM-EDX reveal at least four subsequent stages of quartz crystallization and fragmentation. (i) The oldest generation of quartz is represented by a homogeneous dark grey to reddish quartz mass made up by ~10-20 μm-sized crystals. It contains mm- to cm-sized angular wall-rock fragments, completely altered to kaolinite, indicating intense wall-rock alteration prior to the earliest event of silica precipitation. This rules out the possibility that the quartz mass developed from silicification of the wall rocks. This first type of quartz occurs as cm- to dm-large angular fragments in (ii) a light grey to pink quartz mass formed by ~10-50 μm-sized crystals. The different colours result from variable types and amounts of inclusions. Quartz of both generations shows random crystallographic orientations and complex inclusion structures. It probably developed during two fragmentation events and possibly from a silica gel precursor that crystallized after precipitation. (iii) The third quartz generation formed as a set of mm- to dm-wide veins roughly parallel to the trend of the Pfahl zone

  9. APP Is a Context-Sensitive Regulator of the Hippocampal Presynaptic Active Zone.

    Melanie Laßek

    2016-04-01

    Full Text Available The hallmarks of Alzheimer's disease (AD are characterized by cognitive decline and behavioral changes. The most prominent brain region affected by the progression of AD is the hippocampal formation. The pathogenesis involves a successive loss of hippocampal neurons accompanied by a decline in learning and memory consolidation mainly attributed to an accumulation of senile plaques. The amyloid precursor protein (APP has been identified as precursor of Aβ-peptides, the main constituents of senile plaques. Until now, little is known about the physiological function of APP within the central nervous system. The allocation of APP to the proteome of the highly dynamic presynaptic active zone (PAZ highlights APP as a yet unknown player in neuronal communication and signaling. In this study, we analyze the impact of APP deletion on the hippocampal PAZ proteome. The native hippocampal PAZ derived from APP mouse mutants (APP-KOs and NexCreAPP/APLP2-cDKOs was isolated by subcellular fractionation and immunopurification. Subsequently, an isobaric labeling was performed using TMT6 for protein identification and quantification by high-resolution mass spectrometry. We combine bioinformatics tools and biochemical approaches to address the proteomics dataset and to understand the role of individual proteins. The impact of APP deletion on the hippocampal PAZ proteome was visualized by creating protein-protein interaction (PPI networks that incorporated APP into the synaptic vesicle cycle, cytoskeletal organization, and calcium-homeostasis. The combination of subcellular fractionation, immunopurification, proteomic analysis, and bioinformatics allowed us to identify APP as structural and functional regulator in a context-sensitive manner within the hippocampal active zone network.

  10. APP Is a Context-Sensitive Regulator of the Hippocampal Presynaptic Active Zone.

    Laßek, Melanie; Weingarten, Jens; Wegner, Martin; Mueller, Benjamin F; Rohmer, Marion; Baeumlisberger, Dominic; Arrey, Tabiwang N; Hick, Meike; Ackermann, Jörg; Acker-Palmer, Amparo; Koch, Ina; Müller, Ulrike; Karas, Michael; Volknandt, Walter

    2016-04-01

    The hallmarks of Alzheimer's disease (AD) are characterized by cognitive decline and behavioral changes. The most prominent brain region affected by the progression of AD is the hippocampal formation. The pathogenesis involves a successive loss of hippocampal neurons accompanied by a decline in learning and memory consolidation mainly attributed to an accumulation of senile plaques. The amyloid precursor protein (APP) has been identified as precursor of Aβ-peptides, the main constituents of senile plaques. Until now, little is known about the physiological function of APP within the central nervous system. The allocation of APP to the proteome of the highly dynamic presynaptic active zone (PAZ) highlights APP as a yet unknown player in neuronal communication and signaling. In this study, we analyze the impact of APP deletion on the hippocampal PAZ proteome. The native hippocampal PAZ derived from APP mouse mutants (APP-KOs and NexCreAPP/APLP2-cDKOs) was isolated by subcellular fractionation and immunopurification. Subsequently, an isobaric labeling was performed using TMT6 for protein identification and quantification by high-resolution mass spectrometry. We combine bioinformatics tools and biochemical approaches to address the proteomics dataset and to understand the role of individual proteins. The impact of APP deletion on the hippocampal PAZ proteome was visualized by creating protein-protein interaction (PPI) networks that incorporated APP into the synaptic vesicle cycle, cytoskeletal organization, and calcium-homeostasis. The combination of subcellular fractionation, immunopurification, proteomic analysis, and bioinformatics allowed us to identify APP as structural and functional regulator in a context-sensitive manner within the hippocampal active zone network.

  11. Zeolite H-BEA catalysed multicomponent reaction: One-pot synthesis of amidoalkyl naphthols - Biologically active drug-like molecules

    Sunil R Mistry; Rikesh S Joshi; Kalpana C Maheria

    2011-07-01

    Zeolite has been used as an efficient and a novel heterogeneous catalyst for one-pot synthesis of biologically active drug-like molecules, amidoalkyl naphthols. This green route involves multicomponent reaction of 2-naphthol, aromatic aldehydes and amide in the presence of a catalytic amount of zeolite H-Beta (H-BEA) under solvent reflux as well as solvent-free conditions.

  12. Single molecule activity measurements of cytochrome P450 oxidoreductase reveal the existence of two discrete functional states

    Laursen, Tomas; Singha, Aparajita; Rantzau, Nicolai;

    2014-01-01

    Electron transfer between membrane spanning oxidoreductase enzymes controls vital metabolic processes. Here we studied for the first time with single molecule resolution the function of P450 oxidoreductase (POR), the canonical membrane spanning activator of all microsomal cytochrome P450 enzymes....

  13. Transmembrane α-Helix 2 and 7 Are Important for Small Molecule-Mediated Activation of the GLP-1 Receptor

    Underwood, Christina Rye; Møller Knudsen, Sanne; Schjellerup Wulff, Birgitte;

    2011-01-01

    , the structurally related small molecule, compound 3, stimulated cAMP production from GLP-1R, but not from the homologous glucagon receptor (GluR). The receptor selectivity encouraged a chimeric receptor approach to identify domains important for compound 3-mediated activation of GLP-1R. A subsegment of the GLP-1R...

  14. HLA-DR molecules enhance signal transduction through the CD3/Ti complex in activated T cells

    Odum, Niels; Martin, P J; Schieven, G L;

    1991-01-01

    Crosslinking HLA-DR molecules by monoclonal antibodies (mAb) induces protein tyrosine phosphorylation and results in a secondary elevation of free cytoplasmic Ca2+ concentration ([Ca2+]i) in activated human T cells. Here we have studied the effect of DR on CD3-induced signal transduction...

  15. Significant foreshock activities of M>7.5 earthquakes in the Kuril subduction zone

    Harada, T.; Yokoi, S.; Satake, K.

    2014-12-01

    In the Kuril subduction zone, some M>7.5 earthquakes are accompanied by significant foreshock activities, providing a good opportunity to understand the characteristics of foreshocks for large interplate events such as occur along the Japan Trench and Nankai Trough etc. Some preliminary results from our examination of the foreshock sequences are as follows. Relocated foreshocks tend to migrate with time toward the trench axis. Foreshock distributions of the interplate earthquakes do not overlap with the large coseismic slips (asperities) of the mainshocks. Foreshocks of the 2007 northern Kuril outer-rise event, however, were distributed on the entire rupture area. Foreshock sequences seem to be limited in the regions where the background seismicity rates are relatively high. The foreshock activities were found in the examination of the space-time pattern of M>7 events along the northern Japan to Kuril trench since 1913 (e.g. Harada, Satake, and Ishibashi, 2011:AGU, 2012:AOGS). The large earthquakes preceded by active foreshock sequences are: the 2006 (M8.3), 2007 (M8.1) offshore Simushir earthquakes, the 1963 (M8.5), 1991 (M7.6), 1995 (M7.9) offshore Urup events, the 1978 (M7.8) offshore Iturup events, the 1969 (M8.2) offshore Shikotan event. In contrast, M>7.5 interplate earthquakes offshore Hokkaido (1952 (M8.1), 1973 (M7.8), 2003 (M8.1)) and intraslab earthquakes (1958 (M8.3), 1978 (M7.8), 1993 (M7.6), 1994 (M8.3)) had few or no foreshocks. In the examination of the active foreshocks, we relocated foreshocks by the Modified JHD method (Hurukawa, 1995), compared relocated foreshock areas with mainshock coseismic slip distributions estimated by the teleseismic body-wave inversion (Kikuchi and Kanamori, 2003), and examined the relation between active foreshock sequences and regional background seismicity. This study was supported by the MEXT's "New disaster mitigation research project on Mega thrust earthquakes around Nankai/Ryukyu subduction zones".

  16. The River Network, Active Tectonics and the Mexican Subduction Zone, Southwest Mexico

    Gaidzik, K.; Ramirez-Herrera, M. T.; Kostoglodov, V.; Basili, R.

    2014-12-01

    Rivers, their profiles and network reflect the integration of multiple processes and forces that are part of the fundamental controls on the relief structure of mountain belts. The motivation of this study is to understand active tectonic processes in the forearc region of subduction zones, by distinguishing evidence of active deformation using the river network and topography. To this end, morphotectonic and structural studies have been conducted on fifteen drainage basins on the mountain front, parallel to the Mexican subduction zone, where the Cocos plate underthrusts the North American plate. The southwest - northeast Cocos plate subduction stress regime initiated ca. 20 MA. NE-SW to NNE-SSW normal faults as well as sub-latitudinal to NW-SE strike-slip faults (both dextral and sinistral) constitute the majority of mesofaults recorded in the field within the studied drainage basins. Occasionally dextral N-S strike-slip faults also occur. The stress tensor reconstruction suggests two main evolution stages of these faults: 1) the older is dominated by a NW-SE to WNW-ESE extensional regime and 2) the younger is a transcurrent regime, with NNE-SSW σ1 axis. The drainage pattern is strongly controlled by tectonic features, whereas lithology is only a subordinate factor, with only one exception (Petatlán river). Generally, major rivers flow from north to south mainly through NE-SW and NNE-SSW normal faults, and/or sub-longitudinal dextral (also locally sinistral) strike-slip faults. In the central and eastern part of the studied area, rivers also follow NW-SE structures, which are generally normal or sinistral strike-slip faults (rarely reverse). In most cases, local deflections of the river main courses are related to sub-latitudinal strike-slip faults, both dextral and sinistral. Within the current stress field related to the active Cocos subduction, both normal and strike-slip fault sets could be reactivated. Our analysis suggests that strike-slip faults, mainly

  17. Machine learning models identify molecules active against the Ebola virus in vitro [version 2; referees: 2 approved

    Sean Ekins

    2016-01-01

    Full Text Available The search for small molecule inhibitors of Ebola virus (EBOV has led to several high throughput screens over the past 3 years. These have identified a range of FDA-approved active pharmaceutical ingredients (APIs with anti-EBOV activity in vitro and several of which are also active in a mouse infection model. There are millions of additional commercially-available molecules that could be screened for potential activities as anti-EBOV compounds. One way to prioritize compounds for testing is to generate computational models based on the high throughput screening data and then virtually screen compound libraries. In the current study, we have generated Bayesian machine learning models with viral pseudotype entry assay and the EBOV replication assay data. We have validated the models internally and externally. We have also used these models to computationally score the MicroSource library of drugs to select those likely to be potential inhibitors. Three of the highest scoring molecules that were not in the model training sets, quinacrine, pyronaridine and tilorone, were tested in vitro and had EC50 values of 350, 420 and 230 nM, respectively. Pyronaridine is a component of a combination therapy for malaria that was recently approved by the European Medicines Agency, which may make it more readily accessible for clinical testing. Like other known antimalarial drugs active against EBOV, it shares the 4-aminoquinoline scaffold. Tilorone, is an investigational antiviral agent that has shown a broad array of biological activities including cell growth inhibition in cancer cells, antifibrotic properties, α7 nicotinic receptor agonist activity, radioprotective activity and activation of hypoxia inducible factor-1. Quinacrine is an antimalarial but also has use as an anthelmintic. Our results suggest data sets with less than 1,000 molecules can produce validated machine learning models that can in turn be utilized to identify novel EBOV inhibitors in

  18. Machine learning models identify molecules active against the Ebola virus in vitro [version 1; referees: 2 approved

    Sean Ekins

    2015-10-01

    Full Text Available The search for small molecule inhibitors of Ebola virus (EBOV has led to several high throughput screens over the past 3 years. These have identified a range of FDA-approved active pharmaceutical ingredients (APIs with anti-EBOV activity in vitro and several of which are also active in a mouse infection model. There are millions of additional commercially-available molecules that could be screened for potential activities as anti-EBOV compounds. One way to prioritize compounds for testing is to generate computational models based on the high throughput screening data and then virtually screen compound libraries. In the current study, we have generated Bayesian machine learning models with viral pseudotype entry assay and the EBOV replication assay data. We have validated the models internally and externally. We have also used these models to computationally score the MicroSource library of drugs to select those likely to be potential inhibitors. Three of the highest scoring molecules that were not in the model training sets, quinacrine, pyronaridine and tilorone, were tested in vitro and had EC50 values of 350, 420 and 230 nM, respectively. Pyronaridine is a component of a combination therapy for malaria that was recently approved by the European Medicines Agency, which may make it more readily accessible for clinical testing. Like other known antimalarial drugs active against EBOV, it shares the 4-aminoquinoline scaffold. Tilorone, is an investigational antiviral agent that has shown a broad array of biological activities including cell growth inhibition in cancer cells, antifibrotic properties, α7 nicotinic receptor agonist activity, radioprotective activity and activation of hypoxia inducible factor-1. Quinacrine is an antimalarial but also has use as an anthelmintic. Our results suggest data sets with less than 1,000 molecules can produce validated machine learning models that can in turn be utilized to identify novel EBOV inhibitors in

  19. Impacts of stellar evolution and dynamics on the habitable zone: The role of rotation and magnetic activity

    Gallet, F.; Charbonnel, C.; Amard, L.; Brun, S.; Palacios, A.; Mathis, S.

    2017-01-01

    Context. With the ever growing number of detected and confirmed exoplanets, the probability of finding a planet that looks like the Earth increases continuously. While it is clear that the presence of a planet in the habitable zone does not imply the planet is habitable, a systematic study of the evolution of the habitable zone is required to account for its dependence on stellar parameters. Aims: In this article, we aim to provide the community with the dependence of the habitable zone upon the stellar mass, metallicity, rotation, and for various prescriptions of the limits of the habitable zone. Methods: We use stellar evolution models computed with the code STAREVOL, which includes the most current physical mechanisms of internal transport of angular momentum and external wind braking, to study the evolution of the habitable zone and the continuously habitable zone limits. Results: The stellar parameters mass and metallicity affect the habitable zone limits most dramatically. Conversely, for a given stellar mass and metallicity, stellar rotation has only a marginal effect on these limits and does not modify the width of the habitable zone. Moreover, and as expected in the main-sequence phase and for a given stellar mass and metallicity, the habitable zone limits remain almost constant, and this confirms the usual assumptions of a relative constancy of these limits during that phase. The evolution of the habitable zone limits is also correlated to the evolution of the stellar activity (through the Rossby number), which depends on the stellar mass considered. While the magnetic activity has negligible consequence in the case of more massive stars, these effects may have a strong impact on the habitability of a planet around M-dwarf stars. Thus, stellar activity cannot be neglected and may have a strong impact on the development of life during the early stage of the continuously habitable zone phase of low-mass stars. Using observed trends of stellar magnetic field

  20. Calculations of Induced Activity in the ATLAS Experiment for Nuclear Waste Zoning.

    Morev, M N

    2007-01-01

    Extensive calculations were performed with the general activation formula using the fluxes of high-energy hadrons and low-energy neutrons previously obtained from simulations with the GCALOR code of the ATLAS detector. Three sets of proton cross-sections were used for hadrons energy above 20 MeV: (a) one set calculated with the YIELDX code (i.e., the Silberberg-Tsao formula of partial proton spallation cross-sections), (b) one set calculated with the Rudstam formula, and (c) the ‘best-estimate' dataset which was a compilation of the available experimental and calculated data. In the energy region below 20 MeV, neutron activation cross-sections were taken from evaluated nuclear data files. The activity of each nuclide for a predefined operation scenario (i.e., number and duration of irradiation and shutdown cycles) was normalized to reference values taken from the European or Swiss legislations, to obtain an aggregate estimate of the radiological hazard comparable with a nuclear waste zoning definition cr...

  1. Mechanisms of short-term plasticity at neuromuscular active zones of Drosophila

    Hallermann, Stefan; Heckmann, Manfred; Kittel, Robert J.

    2010-01-01

    During short bursts of neuronal activity, changes in the efficacy of neurotransmitter release are governed primarily by two counteracting processes: (1) Ca2+-dependent elevations of vesicle release probability and (2) depletion of synaptic vesicles. The dynamic interplay of both processes contributes to the expression of activity-dependent synaptic plasticity. Here, we exploited various facets of short-term plasticity at the Drosophila neuromuscular junction to dissect these two processes. This enabled us to rigorously analyze different models of synaptic vesicle pools in terms of their size and mobilization properties. Independent of the specific model, we estimate ∼300 readily releasable vesicles with an average release probability of ∼50% in 1 mM extracellular calcium (∼5% in 0.4 mM extracellular calcium) under resting conditions. The models also helped interpreting the altered short-term plasticity of the previously reported mutant of the active zone component Bruchpilot (BRP). Finally, our results were independently confirmed through fluctuation analysis. Our data reveal that the altered short-term plasticity observed in BRP mutants cannot be accounted for by delocalized Ca2+ channels alone and thus suggest an additional role of BRP in short-term plasticity. PMID:20811513

  2. Underground Corrosion of Activated Metals in an Arid Vadose Zone Environment

    Adler Flitton, Mariana Kay; Mizia, Ronald Eugene; Bishop, Carolyn Wagoner

    2002-04-01

    The subsurface radioactive disposal site located at the Idaho National Engineering and Environmental Laboratory contains neutron-activated metals from nonfuel nuclear-reactor- core components. A long-term corrosion test is being conducted to obtain site-specific corrosion rates to support efforts to more accurately estimate the transfer of activated elements in an arid vadose zone environment. The tests use nonradioactive metal coupons representing the prominent neutron-activated material buried at the disposal location, namely, Type 304L stainless steel, Type 315L stainless steel, nickel-chromium alloy (UNS NO7718), beryllium, aluminum 6061-T6, and a zirconium alloy, (UNS R60804). In addition, carbon steel (the material presently used in the cask disposal liners and other disposal containers) and a duplex stainless steel (UNS S32550) (the proposed material for the high- integrity disposal containers) are also included in the test program. This paper briefly describes the test program and presents the early corrosion rate results after 1 year and 3 years of underground exposure.

  3. Underground Corrosion of Activated Metals in an Arid Vadose Zone Environment

    Adler Flitton, M.K; Mizia, R.E.; Bishop, C.W.

    2001-10-24

    The subsurface radioactive disposal site located at the Idaho National Engineering and Environmental Laboratory contains neutron-activated metals from nonfuel nuclear-reactor- core components. A long-term corrosion test is being conducted to obtain site-specific corrosion rates to support efforts to more accurately estimate the transfer of activated elements in an arid vadose zone environment. The tests use nonradioactive metal coupons representing the prominent neutron-activated material buried at the disposal location, namely, Type 304L stainless steel, Type 315L stainless steel, nickel-chromium alloy (UNS NO7718), beryllium, aluminum 6061-T6, and a zirconium alloy, (UNS R60804). In addition, carbon steel (the material presently used in the cask disposal liners and other disposal containers) and a duplex stainless steel (UNS S32550) (the proposed material for the high- integrity disposal containers) are also included in the test program. This paper briefly describes the test program and presents the early corrosion rate results after 1 year and 3 years of underground exposure.

  4. Interplay of solvent additive concentration and active layer thickness on the performance of small molecule solar cells.

    Love, John A; Collins, Samuel D; Nagao, Ikuhiro; Mukherjee, Subhrangsu; Ade, Harald; Bazan, Guillermo C; Nguyen, Thuc-Quyen

    2014-11-19

    A relationship between solvent additive concentration and active layer thickness in small-molecule solar cells is investigated. Specifically, the additive concentration must scale with the amount of semiconductor material and not as absolute concentration in solution. Devices with a wide range of active layers with thickness up to 200 nm can readily achieve efficiencies close to 6% when the right concentration of additive is used.

  5. Phosphate Activation via Reduced Oxidation State Phosphorus (P. Mild Routes to Condensed-P Energy Currency Molecules

    Claire R. Cousins

    2013-07-01

    Full Text Available The emergence of mechanisms for phosphorylating organic and inorganic molecules is a key step en route to the earliest living systems. At the heart of all contemporary biochemical systems reside reactive phosphorus (P molecules (such as adenosine triphosphate, ATP as energy currency molecules to drive endergonic metabolic processes and it has been proposed that a predecessor of such molecules could have been pyrophosphate [P2O74−; PPi(V]. Arguably the most geologically plausible route to PPi(V is dehydration of orthophosphate, Pi(V, normally a highly endergonic process in the absence of mechanisms for activating Pi(V. One possible solution to this problem recognizes the presence of reactive-P containing mineral phases, such as schreibersite [(Fe,Ni3P] within meteorites whose abundance on the early Earth would likely have been significant during a putative Hadean-Archean heavy bombardment. Here, we propose that the reduced oxidation state P-oxyacid, H-phosphite [HPO32−; Pi(III] could have activated Pi(V towards condensation via the intermediacy of the condensed oxyacid pyrophosphite [H2P2O52−; PPi(III]. We provide geologically plausible provenance for PPi(III along with evidence of its ability to activate Pi(V towards PPi(V formation under mild conditions (80 °C in water.

  6. Phosphate Activation via Reduced Oxidation State Phosphorus (P). Mild Routes to Condensed-P Energy Currency Molecules

    Kee, Terence P.; Bryant, David E.; Herschy, Barry; Marriott, Katie E. R.; Cosgrove, Nichola E.; Pasek, Matthew A.; Atlas, Zachary D.; Cousins, Claire R.

    2013-01-01

    The emergence of mechanisms for phosphorylating organic and inorganic molecules is a key step en route to the earliest living systems. At the heart of all contemporary biochemical systems reside reactive phosphorus (P) molecules (such as adenosine triphosphate, ATP) as energy currency molecules to drive endergonic metabolic processes and it has been proposed that a predecessor of such molecules could have been pyrophosphate [P2O74−; PPi(V)]. Arguably the most geologically plausible route to PPi(V) is dehydration of orthophosphate, Pi(V), normally a highly endergonic process in the absence of mechanisms for activating Pi(V). One possible solution to this problem recognizes the presence of reactive-P containing mineral phases, such as schreibersite [(Fe,Ni)3P] within meteorites whose abundance on the early Earth would likely have been significant during a putative Hadean-Archean heavy bombardment. Here, we propose that the reduced oxidation state P-oxyacid, H-phosphite [HPO32−; Pi(III)] could have activated Pi(V) towards condensation via the intermediacy of the condensed oxyacid pyrophosphite [H2P2O52−; PPi(III)]. We provide geologically plausible provenance for PPi(III) along with evidence of its ability to activate Pi(V) towards PPi(V) formation under mild conditions (80 °C) in water. PMID:25369812

  7. Phosphodiesterase7 Inhibition Activates Adult Neurogenesis in Hippocampus and Subventricular Zone In Vitro and In Vivo.

    Morales-Garcia, Jose A; Echeverry-Alzate, Victor; Alonso-Gil, Sandra; Sanz-SanCristobal, Marina; Lopez-Moreno, Jose A; Gil, Carmen; Martinez, Ana; Santos, Angel; Perez-Castillo, Ana

    2017-02-01

    The phosphodiesterase 7 (PDE7) enzyme is one of the enzymes responsible for controlling intracellular levels of cyclic adenosine 3',5'-monophosphate in the immune and central nervous system. We have previously shown that inhibitors of this enzyme are potent neuroprotective and anti-inflammatory agents. In addition, we also demonstrated that PDE7 inhibition induces endogenous neuroregenerative processes toward a dopaminergic phenotype. Here, we show that PDE7 inhibition controls stem cell expansion in the subgranular zone of the dentate gyrus of the hippocampus (SGZ) and the subventricular zone (SVZ) in the adult rat brain. Neurospheres cultures obtained from SGZ and SVZ of adult rats treated with PDE7 inhibitors presented an increased proliferation and neuronal differentiation compared to control cultures. PDE7 inhibitors treatment of neurospheres cultures also resulted in an increase of the levels of phosphorylated cAMP response element binding protein, suggesting that their effects were indeed mediated through the activation of the cAMP/PKA signaling pathway. In addition, adult rats orally treated with S14, a specific inhibitor of PDE7, presented elevated numbers of proliferating progenitor cells, and migrating precursors in the SGZ and the SVZ. Moreover, long-term treatment with this PDE7 inhibitor shows a significant increase in newly generated neurons in the olfactory bulb and the hippocampus. Also a better performance in memory tests was observed in S14 treated rats, suggesting a functional relevance for the S14-induced increase in SGZ neurogenesis. Taken together, our results indicate for the first time that inhibition of PDE7 directly regulates proliferation, migration and differentiation of neural stem cells, improving spatial learning and memory tasks. Stem Cells 2017;35:458-472.

  8. Conformational, spectroscopic and nonlinear optical properties of biologically active N,N-dimethyltryptamine molecule: A theoretical study

    Öner, Nazmiye; Tamer, Ömer; Avcı, Davut; Atalay, Yusuf

    2014-12-01

    The effective psychoactive properties of N,N-dimethyltryptamine (DMT) known as the near-death molecule have encouraged the imagination of many research disciplines for several decades. Although there is no theoretical study, a number of paper composed by experimental techniques have been reported for DMT molecule. In this study, the molecular modeling of DMT was carried out using B3LYP and HSEh1PBE levels of density functional theory (DFT). Our calculations showed that the energy gap between HOMO and LUMO is low, demonstrating that DMT is a biologically active molecule. Large hyperconjugation interaction energies imply that molecular charge transfer occurs in DMT. Moreover, NLO analysis indicates that DMT can be used an effective NLO material.

  9. [Correlation analysis between meteorological factors, biomass, and active components of Salvia miltiorrhiza in different climatic zones].

    Zhang, Chen-lu; Liang, Zong-suo; Guo, Hong-bo; Liu, Jing-ling; Liu, Yan; Liu, Feng-hua; Wei, Lang-zhu

    2015-02-01

    In this study, the growth and accumulation of active components of Salvia miltiorrhiza in twenty two experimental sites which crossing through three typical climate zones. The S. miltiorrhiza seedlings with the same genotype were planted in each site in spring, which were cultivated in fields with uniform management during their growing seasons till to harvest. The diterpene ketones (dihydrotanshinone, cryptotanshinone, tanshinone I and tanshinone II(A)) in S. miltiorrhiza root samples were determined by using high-performance liquid chromatography (HPLC) method. The biomass of root (root length, number of root branches, root width and dry weight) was also measured. The results showed that tanshinone II(A) in all samples of each site were higher than the standards required by China Pharmacopoeia. It has been found there is a relationship between root shape and climate change. The correlation analysis between active components and meteorological factors showed that the accumulation of tanshinones were effected by such meteorological factors as average relative humidity from April to October > average vapor pressure from April to October > average temperature difference day and night from April to October > annual average temperature and so on. The correlation analysis between root biomass and meteorological factors exhibited that root shape and accumulation of dry matter were affected by those factors, such as average annual aboveground (0-20 cm) temperature from April to October > annual average temperature > average vapor pressure from April to October > annual active accumulated temperature > annual average temperature > average vapor pressure from April to October. The accumulation of tanshinones and biomass was increased with the decrease of latitude. At the same time, the dry matter and diameter of root decreased if altitude rises. In addition, S. miltiorrhiza required sunlight is not sophisticated, when compared with humid and temperature. To sum up, S

  10. Biochemical and toxicological evaluation of nano-heparins in cell functional properties, proteasome activation and expression of key matrix molecules.

    Piperigkou, Zoi; Karamanou, Konstantina; Afratis, Nikolaos A; Bouris, Panagiotis; Gialeli, Chrysostomi; Belmiro, Celso L R; Pavão, Mauro S G; Vynios, Dimitrios H; Tsatsakis, Aristidis M

    2016-01-05

    The glycosaminoglycan heparin and its derivatives act strongly on blood coagulation, controlling the activity of serine protease inhibitors in plasma. Nonetheless, there is accumulating evidence highlighting different anticancer activities of these molecules in numerous types of cancer. Nano-heparins may have great biological significance since they can inhibit cell proliferation and invasion as well as inhibiting proteasome activation. Moreover, they can cause alterations in the expression of major modulators of the tumor microenvironment, regulating cancer cell behavior. In the present study, we evaluated the effects of two nano-heparin formulations: one isolated from porcine intestine and the other from the sea squirt Styela plicata, on a breast cancer cell model. We determined whether these nano-heparins are able to affect cell proliferation, apoptosis and invasion, as well as proteasome activity and the expression of extracellular matrix molecules. Specifically, we observed that nano-Styela compared to nano-Mammalian analogue has higher inhibitory role on cell proliferation, invasion and proteasome activity. Moreover, nano-Styela regulates cell apoptosis, expression of inflammatory molecules, such as IL-6 and IL-8 and reduces the expression levels of extracellular matrix macromolecules, such as the proteolytic enzymes MT1-MMP, uPA and the cell surface proteoglycans syndecan-1 and -2, but not on syndecan-4. The observations reported in the present article indicate that nano-heparins and especially ascidian heparin are effective agents for heparin-induced effects in critical cancer cell functions, providing an important possibility in pharmacological targeting.

  11. Combined single channel and single molecule detection identifies subunit composition of STIM1-activated transient receptor potential canonical (TRPC) channels.

    Asanov, Alexander; Sampieri, Alicia; Moreno, Claudia; Pacheco, Jonathan; Salgado, Alfonso; Sherry, Ryan; Vaca, Luis

    2015-01-01

    Depletion of intracellular calcium ion stores initiates a rapid cascade of events culminating with the activation of the so-called Store-Operated Channels (SOC) at the plasma membrane. Calcium influx via SOC is essential in the initiation of calcium-dependent intracellular signaling and for the refilling of internal calcium stores, ensuring the regeneration of the signaling cascade. In spite of the significance of this evolutionary conserved mechanism, the molecular identity of SOC has been the center of a heated controversy spanning over the last 20 years. Initial studies positioned some members of the transient receptor potential canonical (TRPC) channel superfamily of channels (with the more robust evidence pointing to TRPC1) as a putative SOC. Recent evidence indicates that Stromal Interacting Molecule 1 (STIM1) activates some members from the TRPC family of channels. However, the exact subunit composition of TRPC channels remains undetermined to this date. To identify the subunit composition of STIM1-activated TRPC channels, we developed novel method, which combines single channel electrophysiological measurements based on the patch clamp technique with single molecule fluorescence imaging. We termed this method Single ion Channel Single Molecule Detection technique (SC-SMD). Using SC-SMD method, we have obtained direct evidence of the subunit composition of TRPC channels activated by STIM1. Furthermore, our electrophysiological-imaging SC-SMD method provides evidence at the molecular level of the mechanism by which STIM1 and calmodulin antagonize to modulate TRPC channel activity.

  12. Interaction of an immunodominant epitope with Ia molecules in T-cell activation

    Adorini, L; Sette, A; Buus, S;

    1988-01-01

    The amino acid sequence corresponding to residues 107-116 of hen egg-white lysozyme (HEL) has been identified as containing an immunodominant T-cell epitope recognized in association with the I-Ed molecule. The immunodominance of this epitope in HEL-primed H-2d mice was demonstrated by analysis o......-120)-peptide was found to be immunogenic in H-2d mice. Thus, a single semiconservative substitution drastically reduces binding capacity and abolishes immunogenicity, suggesting that a strict correlation exists between binding of a peptide to Ia molecules and its immunogenicity....

  13. Ground-state kinetics of bistable redox-active donor-acceptor mechanically interlocked molecules.

    Fahrenbach, Albert C; Bruns, Carson J; Li, Hao; Trabolsi, Ali; Coskun, Ali; Stoddart, J Fraser

    2014-02-18

    The ability to design and confer control over the kinetics of theprocesses involved in the mechanisms of artificial molecular machines is at the heart of the challenge to create ones that can carry out useful work on their environment, just as Nature is wont to do. As one of the more promising forerunners of prototypical artificial molecular machines, chemists have developed bistable redox-active donor-acceptor mechanically interlocked molecules (MIMs) over the past couple of decades. These bistable MIMs generally come in the form of [2]rotaxanes, molecular compounds that constitute a ring mechanically interlocked around a dumbbell-shaped component, or [2]catenanes, which are composed of two mechanically interlocked rings. As a result of their interlocked nature, bistable MIMs possess the inherent propensity to express controllable intramolecular, large-amplitude, and reversible motions in response to redox stimuli. In this Account, we rationalize the kinetic behavior in the ground state for a large assortment of these types of bistable MIMs, including both rotaxanes and catenanes. These structures have proven useful in a variety of applications ranging from drug delivery to molecular electronic devices. These bistable donor-acceptor MIMs can switch between two different isomeric states. The favored isomer, known as the ground-state co-conformation (GSCC) is in equilibrium with the less favored metastable state co-conformation (MSCC). The forward (kf) and backward (kb) rate constants associated with this ground-state equilibrium are intimately connected to each other through the ground-state distribution constant, KGS. Knowing the rate constants that govern the kinetics and bring about the equilibration between the MSCC and GSCC, allows researchers to understand the operation of these bistable MIMs in a device setting and apply them toward the construction of artificial molecular machines. The three biggest influences on the ground-state rate constants arise from

  14. M-ficolin, an innate immune defence molecule, binds patterns of acetyl groups and activates complement

    Frederiksen, Pernille Dorthea; Thiel, Steffen; Larsen, Claus Bindslev;

    2005-01-01

    Ficolins play a role in the innate immune defence as pathogen-associated molecular pattern recognition molecules. Three ficolins are found in humans: H-ficolin, L-ficolin and M-ficolin. L-ficolin and H-ficolin circulate in blood in complexes with mannan-binding lectin-associated serine proteases...

  15. Lymphocyte activation by purified HLA-DR molecules fused into autochthonous "stimulating cells".

    Diu, A; Abikar, K; Rode, H N; Gordon, J

    1985-08-01

    Affinity-purified Ia molecules derived from the Daudi cell line were reconstituted into vesicles with Sendai virus envelope glycoproteins. These vesicles inserted into human peripheral leukocytes could induce stimulation of autologous lymphocytes, as measured by thymidine uptake, 6 days later. It is suggested that this method could provide a means to study allostimulation at the molecular level.

  16. Investigation of Chemical Reactivity, Mass Recovery and Biological Activity During Thermal Treatment of DNAPL Source Zones

    2009-10-01

    shallow, relatively homogeneous, unconfined aquifer ...................................................................................2 2-1...Remediation of DNAPL Source Zones (Dr. Rick Johnson, PI) and ER-1423 Large-Scale Physical Models of Thermal Remediation of DNAPL Source Zones in Aquifers ...Standards and Technology NOM natural organic matter OD outside diameter OW a mixed, methanogenic , PCE-to-ethene-dechlorinating PAH

  17. An Active Area Model of Rapid Infiltration Response at Substantial Depth in the Unsaturated Zone

    Mitchell, L.; Nimmo, J. R.

    2011-12-01

    In a porous medium subject to preferential flow, response to surface water infiltration can occur rapidly even at substantial depth in the unsaturated zone. In a ponding experiment at the Idaho National Laboratory (INL) the profile of undisturbed natural soil, seasonally dry at the start, was observed to approach field saturation throughout a 2 meter depth within 6 hours (Nimmo and Perkins, 2007). Traditional use of Richards' equation would require an unrealistically large unsaturated hydraulic conductivity of 40 m/day to capture the observed non-classic wetting behavior. Here we present a model for rapid flow using an active area concept similar to the active fracture model (Liu and others, 1998, WRR 34:2633-2646). The active area concept is incorporated within the preferential flow domain (which allows rapid downward movement) of a dual-domain model that also contains a diffuse-flow domain in which flow can be described by Richards' equation. Development of the active area model is motivated by observation of rapid wetting at substantial depth, as well as a phenomenon in which deep flow is observed before shallow flow. In this model water movement in the preferential domain can be physically conceptualized as laminar flow in free-surface films of constant average thickness. For a given medium, the preferential domain is characterized by an effective areal density (area per unit bulk volume) that describes the free-surface film capacity of the domain as a function of depth. The active area is defined as a portion of the effective areal density that dictates the depth and temporal distribution of domain-exchange and new infiltration within the preferential domain. With the addition of the active area concept, the model is capable of simulating non-diffusive vertical transport patterns. Advantages of the model include simulating rapid response for a variety of infiltration types, including ponding and rain events, as well as modeling relatively rapid aquifer

  18. Guidance on a better integration of aquaculture, fisheries, and other activities in the coastal zone: from tools to practical examples

    Stelzenmüller, V.; Schulze, T.; Gimpel, A.; Bartelings, H.; Bello, E.; Bergh, O.; Bolman, B.; Caetano, M.; Davaasuren, N.; Fabi, G.; Ferreira, J.G.; Gault, J.; Gramolini, R.; Grati, F.; Hamon, K.G.; Jak, R.G.; Kopke, K.; Laurans, M.; Mäkinen, T.; O’Donnell, V.; O’Hagan, A.M.; O’Mahony, C.; Oostenbrugge, van H.; Ramos, J.; Saurel, C.; Sell, A.L.; Silvo, K.; Sinschek, K.; Soma, K.; Stenberg, C.; Taylor, N.; Vale, C.; Vasquez, F.; Verner-Jeffreys, D.W.

    2013-01-01

    This guidance document provides a comprehensive assessment of the conflicts and synergies between fisheries, aquaculture and other activities in the coastal zone in six COEXIST case study areas. It forms deliverable D5.2 of the COEXIST project and synthesises deliverable D5.1, which provides a more

  19. The Status of Secondary School Science Laboratory Activities for Quality Education in Case of Wolaita Zone, Southern Ethiopia

    Zengele, Ashebir Gogile; Alemayehu, Bereket

    2016-01-01

    A high quality science education in primary and secondary schools contributes to developing scientific literacy and would be expected to predispose students to study the enabling sciences at university. The major purpose of this study was to assess the practice and problems in science laboratory activities in the secondary school of Wolaita Zone,…

  20. Synaptophysin 1 Clears Synaptobrevin 2 from the Presynaptic Active Zone to Prevent Short-Term Depression

    Rajit Rajappa

    2016-02-01

    Full Text Available Release site clearance is an important process during synaptic vesicle (SV recycling. However, little is known about its molecular mechanism. Here we identify self-assembly of exocytosed Synaptobrevin 2 (Syb2 and Synaptophysin 1 (Syp1 by homo- and hetero-oligomerization into clusters as key mechanisms mediating release site clearance for preventing cis-SNARE complex formation at the active zone (AZ. In hippocampal neurons from Syp1 knockout mice, neurons expressing a monomeric Syb2 mutant, or after acute block of the ATPase N-ethylmaleimide-sensitive factor (NSF, responsible for cis-SNARE complex disassembly, we found strong frequency-dependent short-term depression (STD, whereas retrieval of Syb2 by compensatory endocytosis was only affected weakly. Defects in Syb2 endocytosis were stimulus- and frequency-dependent, indicating that Syp1 is not essential for Syb2 retrieval, but for its efficient clearance upstream of endocytosis. Our findings identify an SV protein as a release site clearance factor.

  1. Determination of Four Active Ingredients in Vc Yinqiao Tablets by Capillary Zone Electrophoresis with Amperometric Detection

    L(U),Jin; WANG,Qing-Jiang; CHENG,Xi; LIU,Hai-Yan; HE,Pin-Gang; FANG,Yu-Zhi

    2006-01-01

    A simple, reliable and reproducible method, based on capillary zone electrophoresis with amperometric detection (CZE-AD), has been developed for simultaneous determination of four active ingredients in Vc Yinqiao tablets including paracetamol, vitamin C, caffeic acid and chlorogenic acid. A carbon-disk electrode was used as working electrode and 0.95 V (versus SCE) was selected as detection potential. The optimal conditions of CZE experiment were 30 mmol·L-1 borate solution (pH 9.5) as running buffer, 14 kV as separation voltage and 8 s (14 kV) as electro-kinetic sampling time. Under the selected optimum conditions, paracetamol, vitamin C, caffeic acid and chlorogenic acid could be perfectly separated within 22 min, and their detection limits (S/N=3) ranged from 5 × 10-7 to 1×10-6 mol·L-1. This proposed method demonstrated good reproducibility with relative standard deviations of less than 3% for both migration time and peak current (n=7). The utility of this method was demonstrated by monitoring a kind of compound medicine named Vc Yinqiao tablets and the assay results were satisfactory.

  2. APP-A Novel Player within the Presynaptic Active Zone Proteome.

    Weingarten, Jens; Weingarten, Melanie; Wegner, Martin; Volknandt, Walter

    2017-01-01

    The amyloid precursor protein (APP) was discovered in the 1980s as the precursor protein of the amyloid A4 peptide. The amyloid A4 peptide, also known as A-beta (Aβ), is the main constituent of senile plaques implicated in Alzheimer's disease (AD). In association with the amyloid deposits, increasing impairments in learning and memory as well as the degeneration of neurons especially in the hippocampus formation are hallmarks of the pathogenesis of AD. Within the last decades much effort has been expended into understanding the pathogenesis of AD. However, little is known about the physiological role of APP within the central nervous system (CNS). Allocating APP to the proteome of the highly dynamic presynaptic active zone (PAZ) identified APP as a novel player within this neuronal communication and signaling network. The analysis of the hippocampal PAZ proteome derived from APP-mutant mice demonstrates that APP is tightly embedded in the underlying protein network. Strikingly, APP deletion accounts for major dysregulation within the PAZ proteome network. Ca(2+)-homeostasis, neurotransmitter release and mitochondrial function are affected and resemble the outcome during the pathogenesis of AD. The observed changes in protein abundance that occur in the absence of APP as well as in AD suggest that APP is a structural and functional regulator within the hippocampal PAZ proteome. Within this review article, we intend to introduce APP as an important player within the hippocampal PAZ proteome and to outline the impact of APP deletion on individual PAZ proteome subcommunities.

  3. Trio, a Rho Family GEF, Interacts with the Presynaptic Active Zone Proteins Piccolo and Bassoon

    Terry-Lorenzo, Ryan T.; Torres, Viviana I.; Wagh, Dhananjay; Galaz, Jose; Swanson, Selene K.; Florens, Laurence; Washburn, Michael P.; Waites, Clarissa L.; Gundelfinger, Eckart D.; Reimer, Richard J.; Garner, Craig C.

    2016-01-01

    Synaptic vesicles (SVs) fuse with the plasma membrane at a precise location called the presynaptic active zone (AZ). This fusion is coordinated by proteins embedded within a cytoskeletal matrix assembled at the AZ (CAZ). In the present study, we have identified a novel binding partner for the CAZ proteins Piccolo and Bassoon. This interacting protein, Trio, is a member of the Dbl family of guanine nucleotide exchange factors (GEFs) known to regulate the dynamic assembly of actin and growth factor dependent axon guidance and synaptic growth. Trio was found to interact with the C-terminal PBH 9/10 domains of Piccolo and Bassoon via its own N-terminal Spectrin repeats, a domain that is also critical for its localization to the CAZ. Moreover, our data suggest that regions within the C-terminus of Trio negatively regulate its interactions with Piccolo/Bassoon. These findings provide a mechanism for the presynaptic targeting of Trio and support a model in which Piccolo and Bassoon play a role in regulating neurotransmission through interactions with proteins, including Trio, that modulate the dynamic assembly of F-actin during cycles of synaptic vesicle exo- and endocytosis. PMID:27907191

  4. Active zone scaffolds differentially accumulate Unc13 isoforms to tune Ca(2+) channel-vesicle coupling.

    Böhme, Mathias A; Beis, Christina; Reddy-Alla, Suneel; Reynolds, Eric; Mampell, Malou M; Grasskamp, Andreas T; Lützkendorf, Janine; Bergeron, Dominique Dufour; Driller, Jan H; Babikir, Husam; Göttfert, Fabian; Robinson, Iain M; O'Kane, Cahir J; Hell, Stefan W; Wahl, Markus C; Stelzl, Ulrich; Loll, Bernhard; Walter, Alexander M; Sigrist, Stephan J

    2016-10-01

    Brain function relies on fast and precisely timed synaptic vesicle (SV) release at active zones (AZs). Efficacy of SV release depends on distance from SV to Ca(2+) channel, but molecular mechanisms controlling this are unknown. Here we found that distances can be defined by targeting two unc-13 (Unc13) isoforms to presynaptic AZ subdomains. Super-resolution and intravital imaging of developing Drosophila melanogaster glutamatergic synapses revealed that the Unc13B isoform was recruited to nascent AZs by the scaffolding proteins Syd-1 and Liprin-α, and Unc13A was positioned by Bruchpilot and Rim-binding protein complexes at maturing AZs. Unc13B localized 120 nm away from Ca(2+) channels, whereas Unc13A localized only 70 nm away and was responsible for docking SVs at this distance. Unc13A(null) mutants suffered from inefficient, delayed and EGTA-supersensitive release. Mathematical modeling suggested that synapses normally operate via two independent release pathways differentially positioned by either isoform. We identified isoform-specific Unc13-AZ scaffold interactions regulating SV-Ca(2+)-channel topology whose developmental tightening optimizes synaptic transmission.

  5. Negative regulation of active zone assembly by a newly identified SR protein kinase.

    Ervin L Johnson

    2009-09-01

    Full Text Available Presynaptic, electron-dense, cytoplasmic protrusions such as the T-bar (Drosophila or ribbon (vertebrates are believed to facilitate vesicle movement to the active zone (AZ of synapses throughout the nervous system. The molecular composition of these structures including the T-bar and ribbon are largely unknown, as are the mechanisms that specify their synapse-specific assembly and distribution. In a large-scale, forward genetic screen, we have identified a mutation termed air traffic controller (atc that causes T-bar-like protein aggregates to form abnormally in motoneuron axons. This mutation disrupts a gene that encodes for a serine-arginine protein kinase (SRPK79D. This mutant phenotype is specific to SRPK79D and is not secondary to impaired kinesin-dependent axonal transport. The srpk79D gene is neuronally expressed, and transgenic rescue experiments are consistent with SRPK79D kinase activity being necessary in neurons. The SRPK79D protein colocalizes with the T-bar-associated protein Bruchpilot (Brp in both the axon and synapse. We propose that SRPK79D is a novel T-bar-associated protein kinase that represses T-bar assembly in peripheral axons, and that SRPK79D-dependent repression must be relieved to facilitate site-specific AZ assembly. Consistent with this model, overexpression of SRPK79D disrupts AZ-specific Brp organization and significantly impairs presynaptic neurotransmitter release. These data identify a novel AZ-associated protein kinase and reveal a new mechanism of negative regulation involved in AZ assembly. This mechanism could contribute to the speed and specificity with which AZs are assembled throughout the nervous system.

  6. Activation of Small Molecules by DyI_2 and Dy

    2007-01-01

    1 Results The reactivities of dysprosium diiodide and metallic dysprosium toward small molecules are discussed.For instance,DyI2-induced silyl radical reactions are described.The combination of dysprosium diiodide and dichloromethane can serve as an effective methylene transfer reagent for cyclopropanation of unfunctionalized alkenes beyond that possible with other metal-dichloromethane systems.Furthermore,we report that the combination of chlorosilane and metallic Dy can also serve as an effective prom...

  7. Catalytic Activation of Small Molecules. Development and Characterisation of Ruthenium Complexes for Application in Catalysis

    Choi, Jong-Hoo

    2016-01-01

    In this work, the synthesis, characterisation and catalytic application of ruthenium pincer complexes is presented. In this context, new synthetic strategies are discussed to obtain novel ruthenium pincer dihydrogen complexes. Furthermore, the reactivity of the complexes towards small molecules (e.g. alcohols, boranes, ammonia, amines, nitriles and hydrogen) was observed, delivering fundamental insights into catalytic applications. With the reactivity testing, new borylated B-H-σ-complexes we...

  8. Novel High-Activity Organic Piezoelectric Materials - From Single-Molecule Response to Energy Harvesting Films

    2015-08-24

    1998; xx-xx-1998. 2. REPORT TYPE. State the type of report, such as final, technical, interim, memorandum, master’s thesis , progress, quarterly... Chemistry University of Pittsburgh 219 Parkman Avenue Pittsburgh, PA 15260 E-mail: geoffh@pitt.edu Phone: 412-648-0492 Submitted to Program... inorganic materials. Instead, we designed molecular springs based on helical conjugated molecules with predicted piezo-response and generated

  9. Structures of Clostridium Botulinum Neurotoxin Serotype A Light Chain Complexed with Small-Molecule Inhibitors Highlight Active-Site Flexibility

    Silvaggi,N.; Boldt, G.; Hixon, M.; Kennedy, J.; Tzipori, S.; Janda, K.; Allen, K.

    2007-01-01

    The potential for the use of Clostridial neurotoxins as bioweapons makes the development of small-molecule inhibitors of these deadly toxins a top priority. Recently, screening of a random hydroxamate library identified a small-molecule inhibitor of C. botulinum Neurotoxin Serotype A Light Chain (BoNT/A-LC), 4-chlorocinnamic hydroxamate, a derivative of which has been shown to have in vivo efficacy in mice and no toxicity. We describe the X-ray crystal structures of BoNT/A-LC in complexes with two potent small-molecule inhibitors. The structures of the enzyme with 4-chlorocinnamic hydroxamate or 2,4-dichlorocinnamic hydroxamate bound are compared to the structure of the enzyme complexed with L-arginine hydroxamate, an inhibitor with modest affinity. Taken together, this suite of structures provides surprising insights into the BoNT/A-LC active site, including unexpected conformational flexibility at the S1' site that changes the electrostatic environment of the binding pocket. Information gained from these structures will inform the design and optimization of more effective small-molecule inhibitors of BoNT/A-LC.

  10. Activated peripheral lymphocytes with increased expression of cell adhesion molecules and cytotoxic markers are associated with dengue fever disease

    Elzinandes L Azeredo

    2006-06-01

    Full Text Available The immune mechanisms involved in dengue fever and dengue hemorrhagic/dengue shock syndrome are not well understood. The ex vivo activation status of immune cells during the dengue disease in patients was examined. CD4and CD8 T cells were reduced during the acute phase. Interestingly, CD8 T cells co-expressing activation marker HLA-DR, Q, P, and cytolytic granule protein-Tia-1 were significantly higher in dengue patients than in controls. Detection of adhesion molecules indicated that in dengue patients the majority of T cells (CD4 and CD8 express the activation/memory phenotype, characterized as CD44HIGH and lack the expression of the naïve cell marker, CD62L LOW. Also, the levels of T cells co-expressing ICAM-1 (CD54, VLA-4, and LFA-1 (CD11a were significantly increased. CD8 T lymphocytes expressed predominantly low levels of anti-apoptotic molecule Bcl-2 in the acute phase, possibly leading to the exhibition of a phenotype of activated/effector cells. Circulating levels of IL-18, TGF-b1 and sICAM-1 were significantly elevated in dengue patients. Early activation events occur during acute dengue infection which might contribute to viral clearance. Differences in expression of adhesion molecules among CD4 and CD8 T cells might underlie the selective extravasation of these subsets from blood circulation into lymphoid organs and/or tissues. In addition, activated CD8 T cells would be more susceptible to apoptosis as shown by the alteration in Bcl-2 expression. Cytokines such as IL-18, TGF-b1, and sICAM-1 may be contributing by either stimulating or suppressing the adaptative immune response, during dengue infection, thereby perhaps establishing a relationship with disease severity.

  11. Activated peripheral lymphocytes with increased expression of cell adhesion molecules and cytotoxic markers are associated with dengue fever disease.

    Azeredo, Elzinandes L; Zagne, Sonia M O; Alvarenga, Allan R; Nogueira, Rita M R; Kubelka, Claire F; de Oliveira-Pinto, Luzia M

    2006-06-01

    The immune mechanisms involved in dengue fever and dengue hemorrhagic/dengue shock syndrome are not well understood. The ex vivo activation status of immune cells during the dengue disease in patients was examined. CD4 and CD8 T cells were reduced during the acute phase. Interestingly, CD8 T cells co-expressing activation marker HLA-DR, Q, P, and cytolytic granule protein-Tia-1 were significantly higher in dengue patients than in controls. Detection of adhesion molecules indicated that in dengue patients the majority of T cells (CD4 and CD8) express the activation/memory phenotype, characterized as CD44HIGH and lack the expression of the naïve cell marker, CD62L LOW. Also, the levels of T cells co-expressing ICAM-1 (CD54), VLA-4, and LFA-1 (CD11a) were significantly increased. CD8 T lymphocytes expressed predominantly low levels of anti-apoptotic molecule Bcl-2 in the acute phase, possibly leading to the exhibition of a phenotype of activated/effector cells. Circulating levels of IL-18, TGF-b1 and sICAM-1 were significantly elevated in dengue patients. Early activation events occur during acute dengue infection which might contribute to viral clearance. Differences in expression of adhesion molecules among CD4 and CD8 T cells might underlie the selective extravasation of these subsets from blood circulation into lymphoid organs and/or tissues. In addition, activated CD8 T cells would be more susceptible to apoptosis as shown by the alteration in Bcl-2 expression. Cytokines such as IL-18, TGF-b1, and sICAM-1 may be contributing by either stimulating or suppressing the adaptative immune response, during dengue infection, thereby perhaps establishing a relationship with disease severity.

  12. 78 FR 48413 - Foreign-Trade Zone 75-Phoenix, Arizona, Authorization of Production Activity, Orbital Sciences...

    2013-08-08

    ..., Orbital Sciences Corporation, (Satellites and Spacecraft Launch Vehicles); Gilbert, Arizona On April 2... to the Foreign-Trade Zones (FTZ) Board on behalf of Orbital Sciences Corporation, within Site 10, in... Secretary. BILLING CODE 3510-DS-P...

  13. Auroral Electrojet (AE, AL, AO, AU) - A Global Measure of Auroral Zone Magnetic Activity

    National Oceanic and Atmospheric Administration, Department of Commerce — The AE index is derived from geomagnetic variations in the horizontal component observed at selected (10-13) observatories along the auroral zone in the northern...

  14. Near infrared emission from molecule-like silver clusters confined in zeolite A assisted by thermal activation

    Lin, Hui, E-mail: linh8112@163.com; Imakita, Kenji; Rong Gui, Sa Chu; Fujii, Minoru, E-mail: fujii@eedept.kobe-u.ac.jp [Department of Electrical and Electronic Engineering, Graduate School of Engineering, Kobe University, Rokkodai, Nada, Kobe 657-8501 (Japan)

    2014-07-07

    Strong and broad near infrared (NIR) emission peaked at ~855 nm upon optimal excitation at 342 nm has been observed from molecule-like silver clusters (MLSCs) confined in zeolite A assisted by thermal activation. To the best of our knowledge, this is the first observation of NIR emission peaked at longer than 800 nm from MLSCs confined in solid matrices. The decay time of the NIR emission is over 10 μs, which indicates that it is a spin-forbidden transition. The ~855 nm NIR emission shows strong dependence on the silver loading concentration and the thermal activation temperature.

  15. Exoplanet detection. Stellar activity masquerading as planets in the habitable zone of the M dwarf Gliese 581.

    Robertson, Paul; Mahadevan, Suvrath; Endl, Michael; Roy, Arpita

    2014-07-25

    The M dwarf star Gliese 581 is believed to host four planets, including one (GJ 581d) near the habitable zone that could possibly support liquid water on its surface if it is a rocky planet. The detection of another habitable-zone planet--GJ 581g--is disputed, as its significance depends on the eccentricity assumed for d. Analyzing stellar activity using the Hα line, we measure a stellar rotation period of 130 ± 2 days and a correlation for Hα modulation with radial velocity. Correcting for activity greatly diminishes the signal of GJ 581d (to 1.5 standard deviations) while significantly boosting the signals of the other known super-Earth planets. GJ 581d does not exist, but is an artifact of stellar activity which, when incompletely corrected, causes the false detection of planet g.

  16. Discovery of a small-molecule binder of the oncoprotein gankyrin that modulates gankyrin activity in the cell

    Chattopadhyay, Anasuya; O'Connor, Cornelius J.; Zhang, Fengzhi; Galvagnion, Celine; Galloway, Warren R. J. D.; Tan, Yaw Sing; Stokes, Jamie E.; Rahman, Taufiq; Verma, Chandra; Spring, David R.; Itzhaki, Laura S.

    2016-04-01

    Gankyrin is an ankyrin-repeat oncoprotein whose overexpression has been implicated in the development of many cancer types. Elevated gankyrin levels are linked to aberrant cellular events including enhanced degradation of tumour suppressor protein p53, and inhibition of gankyrin activity has therefore been identified as an attractive anticancer strategy. Gankyrin interacts with several partner proteins, and a number of these protein-protein interactions (PPIs) are of relevance to cancer. Thus, molecules that bind the PPI interface of gankyrin and interrupt these interactions are of considerable interest. Herein, we report the discovery of a small molecule termed cjoc42 that is capable of binding to gankyrin. Cell-based experiments demonstrate that cjoc42 can inhibit gankyrin activity in a dose-dependent manner: cjoc42 prevents the decrease in p53 protein levels normally associated with high amounts of gankyrin, and it restores p53-dependent transcription and sensitivity to DNA damage. The results represent the first evidence that gankyrin is a “druggable” target with small molecules.

  17. Recent advances in small organic molecules as DNA intercalating agents: synthesis, activity, and modeling.

    Rescifina, Antonio; Zagni, Chiara; Varrica, Maria Giulia; Pistarà, Venerando; Corsaro, Antonino

    2014-03-03

    The interaction of small molecules with DNA plays an essential role in many biological processes. As DNA is often the target for majority of anticancer and antibiotic drugs, study about the interaction of drug and DNA has a key role in pharmacology. Moreover, understanding the interactions of small molecules with DNA is of prime significance in the rational design of more powerful and selective anticancer agents. Two of the most important and promising targets in cancer chemotherapy include DNA alkylating agents and DNA intercalators. For these last the DNA recognition is a critical step in their anti-tumor action and the intercalation is not only one kind of the interactions in DNA recognition but also a pivotal step of several clinically used anti-tumor drugs such as anthracyclines, acridines and anthraquinones. To push clinical cancer therapy, the discovery of new DNA intercalators has been considered a practical approach and a number of intercalators have been recently reported. The intercalative binding properties of such molecules can also be harnessed as diagnostic probes for DNA structure in addition to DNA-directed therapeutics. Moreover, the problem of intercalation site formation in the undistorted B-DNA of different length and sequence is matter of tremendous importance in molecular modeling studies and, nowadays, three models of DNA intercalation targets have been proposed that account for the binding features of intercalators. Finally, despite DNA being an important target for several drugs, most of the docking programs are validated only for proteins and their ligands. Therefore, a default protocol to identify DNA binding modes which uses a modified canonical DNA as receptor is needed.

  18. Communities on the move: Pedestrian-oriented zoning as a facilitator of adult active travel to work in the United States

    Jamie Friedman Chriqui

    2016-04-01

    Full Text Available Background: Communities across the United States have been reforming their zoning codes to create pedestrian-friendly neighborhoods with increased street connectivity, mixed-use and higher density, open space, transportation infrastructure, and a traditional neighborhood structure. Zoning code reforms include new urbanist zoning such as the SmartCode, form-based codes, transects, transportation and pedestrian-oriented developments, and traditional neighborhood developments.Purpose: To examine the relationship of zoning code reforms and more active living-oriented zoning provisions with adult active travel to work via walking, biking, or by using public transit.Methods: Zoning codes effective as of 2010 were compiled for 3,914 municipal-level jurisdictions located in 471 counties and 2 consolidated cities in 48 states and the District of Columbia, and that collectively covered 72.9% of the U.S. population. Zoning codes were evaluated for the presence of code reform zoning and nine pedestrian-oriented zoning provisions (1=yes: sidewalks, crosswalks, bike-pedestrian connectivity, street connectivity, bike lanes, bike parking, bike-pedestrian trails/paths, mixed use development, and other walkability/pedestrian-orientation. A zoning scale reflected the number of provisions addressed (out of 10. Five continuous outcome measures were constructed using 2010-2014 American Community Survey municipal-level 5-year estimates to assess the percentage of workers: walking, biking, walking or biking, or taking public transit to work OR engaged in any active travel to work. Regression models controlled for municipal-level socioeconomic characteristics and a GIS-constructed walkability scale and were clustered on county with robust standard errors. Results: Adjusted models indicated that several pedestrian-oriented zoning provisions were statistically associated (p<.05 or lower with increased rates of walking, biking, or engaging in any active travel (walking

  19. A new multiplexing single molecule technique for measuring restriction enzyme activity

    Harbottle, Allison; Cavanaugh, Jillian; Gordon, Wendy; Loparo, Joseph; Price, Allen

    2012-02-01

    We present a new multiplexing single molecule method for observing the cleavage of DNAs by restriction enzymes. DNAs are attached to a surface at one end using a biotin-streptavidin link and to a micro bead at the other end via a digoxigenin-antidigoxigenin link. The DNAs are stretched by applying a flow. After introduction of the restriction enzyme, the exact time of cleavage of individual DNAs is recorded with video microscopy. We can image hundreds to thousands of DNAs in a single experiment. We are using our technique to search for the signature of facilitated diffusion in the measured rate dependence on ionic strength.

  20. Circadian expression of the presynaptic active zone protein Bruchpilot in the lamina of Drosophila melanogaster.

    Górska-Andrzejak, Jolanta; Makuch, Renata; Stefan, Joanna; Görlich, Alicja; Semik, Danuta; Pyza, Elzbieta

    2013-01-01

    In the fly's visual system, the morphology of cells and the number of synapses change during the day. In the present study we show that in the first optic neuropil (lamina) of Drosophila melanogaster, a presynaptic active zone protein Bruchpilot (BRP) exhibits a circadian rhythm in abundance. In day/night (or light/dark, LD) conditions the level of BRP increases two times, in the morning and in the evening. The same pattern of changes in the BRP level was detected in whole brain homogenates, thus indicating that the majority of synapses in the brain peaks twice during the day. However, these two peaks in BRP abundance, measured as the fluorescence intensity of immunolabeling, seem to be regulated differently. The peak in the morning is predominantly regulated by light and involves the transduction pathway in the retina photoreceptors. This peak is present neither in wild-type Canton-S flies in constant darkness (DD), nor in norpA(7) phototransduction mutant in LD. However, it also depends on the clock gene per, because it is abolished in the per(0) arrhythmic mutant. In turn, the peak of BRP in the evening is endogenously regulated by an input from the pacemaker located in the brain. This peak is present in Canton-S flies in DD, as well as in the norpA(7) mutant in LD, but is absent in per(01), tim,(01) and cry(01) mutants in LD. In addition both peaks seem to depend on clock gene-expressing photoreceptors and glial cells of the visual system.

  1. Aftershocks illuminate the 2011 Mineral, Virginia, earthquake causative fault zone and nearby active faults

    Horton, Jr., J. Wright; Shah, Anjana K.; McNamara, Daniel E.; Snyder, Stephen L.; Carter, Aina M

    2015-01-01

    Deployment of temporary seismic stations after the 2011 Mineral, Virginia (USA), earthquake produced a well-recorded aftershock sequence. The majority of aftershocks are in a tabular cluster that delineates the previously unknown Quail fault zone. Quail fault zone aftershocks range from ~3 to 8 km in depth and are in a 1-km-thick zone striking ~036° and dipping ~50°SE, consistent with a 028°, 50°SE main-shock nodal plane having mostly reverse slip. This cluster extends ~10 km along strike. The Quail fault zone projects to the surface in gneiss of the Ordovician Chopawamsic Formation just southeast of the Ordovician–Silurian Ellisville Granodiorite pluton tail. The following three clusters of shallow (illuminate other faults. (1) An elongate cluster of early aftershocks, ~10 km east of the Quail fault zone, extends 8 km from Fredericks Hall, strikes ~035°–039°, and appears to be roughly vertical. The Fredericks Hall fault may be a strand or splay of the older Lakeside fault zone, which to the south spans a width of several kilometers. (2) A cluster of later aftershocks ~3 km northeast of Cuckoo delineates a fault near the eastern contact of the Ordovician Quantico Formation. (3) An elongate cluster of late aftershocks ~1 km northwest of the Quail fault zone aftershock cluster delineates the northwest fault (described herein), which is temporally distinct, dips more steeply, and has a more northeastward strike. Some aftershock-illuminated faults coincide with preexisting units or structures evident from radiometric anomalies, suggesting tectonic inheritance or reactivation.

  2. Gold nano-island arrays on silicon as SERS active substrate for organic molecule detection

    Ignat, Teodora, E-mail: teodora.ignat@imt.ro [National Institute for Research and Development in Microtechnologies, Laboratory of Nanobiotechnology, 126A, Erou Iancu Nicolae Street, 077190 (Romania); Husanu, Marius-Adrian, E-mail: adrianhusanu@gmail.com [National Institute of Materials Physics, Atomistilor Str. 105bis, PO Box MG 7, Magurele, Bucharest 077125 (Romania); Munoz, Roberto, E-mail: rmunoz@icmm.csic.es [Inasmet Fdn, Dept. Biomat and Nanotechnol, San Sebastian (Spain); Kusko, Mihaela, E-mail: mihaela.kusko@imt.ro [National Institute for Research and Development in Microtechnologies, Laboratory of Nanobiotechnology, 126A, Erou Iancu Nicolae Street, 077190 (Romania); Danila, Mihai, E-mail: mihai.danila@imt.ro [National Institute for Research and Development in Microtechnologies, Laboratory of Nanobiotechnology, 126A, Erou Iancu Nicolae Street, 077190 (Romania); Teodorescu, Cristian Mihail, E-mail: teodorescu@infim.ro [National Institute of Materials Physics, Atomistilor Str. 105bis, PO Box MG 7, Magurele, Bucharest 077125 (Romania)

    2014-01-01

    Gold islands forming highly controlled arrays have been fabricated by two potential step electrochemical deposition method using nanopatterned Si surface templates. In the present work, the Raman scattering studies realized using 11-mercaptoundecanoic probe molecule showed that such structures exhibit an enhanced Raman signal compared with nanostructured physical deposited thin gold film on flat silicon substrate and can be valued as surface-enhanced Raman scattering substrates. Besides the more appropriate management of nano-island arrays distribution, the high ratio of their Raman signals can be explain by the epitaxial-like growth mechanism of the metallic nano-islands, clearly showed by X-ray diffraction studies. Furthermore, the substrates enabled reproducibility and stability detection due to the chemically assembling of organothiol molecules, the X-ray photoelectron spectroscopy studies confirming formation of the thiolate species which corresponds to Au-S bonds, and also, the unwanted ‘hot-spots’ are missing, which make them suitable for high sensitivity biosensing applications. - Highlights: • Gold nano-islands are electrochemical deposited on nanopatterned silicon. • The X-ray diffraction studies revealed the epitaxial-like growth mechanism. • Enhanced Raman signal of Au nano-islands was observed compared with Au nano-film.

  3. Neural progenitor cells isolated from the subventricular zone present hemichannel activity and form functional gap junctions with glial cells

    Rocío eTalaverón

    2015-10-01

    Full Text Available The postnatal subventricular zone lining the walls of the lateral ventricles contains neural progenitor cells (NPCs that generate new olfactory bulb interneurons. Communication via gap junctions between cells in the subventricular zone is involved in NPC proliferation and in neuroblast migration towards the olfactory bulb. Subventricular zone NPCs can be expanded in vitro in the form of neurospheres that can be used for transplantation purposes after brain injury. We have previously reported that neurosphere-derived NPCs form heterocellular gap junctions with host glial cells when they are implanted after mechanical injury. To analyze functionality of NPC-glial cell gap junctions we performed dye coupling experiments in co-cultures of subventricular zone NPCs with astrocytes or microglia. Neurosphere-derived cells expressed mRNA for at least the hemichannel/gap junction channel proteins connexin 26 (Cx26, Cx43, Cx45 and pannexin 1. Dye coupling experiments revealed that gap junctional communication occurred among neurosphere cells (incidence of coupling: 100%. Moreover, hemichannel activity was also detected in neurosphere cells as evaluated in time-lapse measurements of ethidium bromide uptake. Heterocellular coupling between NPCs and glial cells was evidenced in co-cultures of neurospheres with astrocytes (incidence of coupling: 91.0 ± 4.7% or with microglia (incidence of coupling: 71.9 ± 6.7%. Dye coupling in neurospheres and in co-cultures was inhibited by octanol, a gap junction blocker. Altogether, these results suggest the existence of functional hemichannels and gap junction channels in postnatal subventricular zone neurospheres. In addition, they demonstrate that subventricular zone-derived NPCs can establish functional gap junctions with astrocytes or microglia. Therefore, cell-cell communication via gap junctions and hemichannels with host glial cells might subserve a role in the functional integration of NPCs after implantation in

  4. [Impacts of root-zone hypoxia stress on muskmelon growth, its root respiratory metabolism, and antioxidative enzyme activities].

    Liu, Yi-Ling; Li, Tian-Lai; Sun, Zhou-Ping; Chen, Ya-Dong

    2010-06-01

    By using aeroponics culture system, this paper studied the impacts of root-zone hypoxia (10% O2 and 5% O2) stress on the plant growth, root respiratory metabolism, and antioxidative enzyme activities of muskmelon at its fruit development stage. Root-zone hypoxia stress inhibited the plant growth of muskmelon, resulting in the decrease of plant height, root length, and fresh and dry biomass. Comparing with the control (21% O2), hypoxia stress reduced the root respiration rate and malate dehydrogenase (MDH) activity significantly, and the impact of 5% O2 stress was more serious than that of 10% O2 stress. Under hypoxic conditions, the lactate dehydrogenase (LDH), alcohol dehydrogenase (ADH), pyruvate decarboxylase (PDC), superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activities and the malondialdehyde (MDA) content were significantly higher than the control. The increment of antioxidative enzyme activities under 10% O2 stress was significantly higher than that under 5% O2 stress, while the MDA content was higher under 5% O2 stress than under 10% O2 stress, suggesting that when the root-zone oxygen concentration was below 10%, the aerobic respiration of muskmelon at its fruit development stage was obviously inhibited while the anaerobic respiration was accelerated, and the root antioxidative enzymes induced defense reaction. With the increasing duration of hypoxic stress, the lipid peroxidation would be aggravated, resulting in the damages on muskmelon roots, inhibition of plant growth, and decrease of fruit yield and quality.

  5. Activation of CO{sub 2} and related small molecules by neopentyl-derivatized uranium complexes

    Schmidt, Anna-Corina

    2015-06-18

    This work reports the newly synthesized neopentyl derivatized tris(aryloxide) U{sup III} complex [(({sup nP,Me}ArO){sub 3}tacn)U{sup III}] (1) and its reactivity with small molecules like nitrous oxide (N{sub 2}O), carbon dioxide (CO{sub 2}), and sulfur dioxide (SO{sub 2}). Additionally, a deeper insight into covalency of U-R bonds with R = O, N and the participation of the f-orbitals to bonding are discussed. For this purpose, a large number of characterization methods were used, such as X-ray diffraction analysis, U{sup V}/vis/NIR, IR vibrational, Raman, X-ray absorption, EPR, and {sup 1}H, {sup 15}N, {sup 13}C and {sup 19}F NMR spectroscopy, cyclic voltammetry, SQUID magnetization measurements and DFT calculations. Moreover, all compounds were checked for purity by elemental analysis.

  6. The range of excursion of flexor tendons in Zone V: a comparison of active vs passive flexion mobilisation regimes.

    Panchal, J

    1997-10-01

    A number of early postoperative mobilisation regimes have been developed in an attempt to increase tendon excursion and gliding and thereby reduce formation of adhesions following repair of flexor tendons. Early active flexion mobilisation regimes are becoming more popular, and have replaced early passive flexion regimes in many centres. The aim of the present study was: (a) to determine the range of excursion of flexor tendons in Zone V, and (b) to compare the excursion ranges between active (Belfast) and passive (modified Duran) flexion mobilisation regimes postoperatively. This was done (a) in two cadavers, and (b) in two patients intraoperatively, and postoperatively at 10 days, 3 weeks and 6 weeks. With passive flexion, the mean tendon excursion in Zone V in cadavers was 1 mm for flexor digitorum superficialis (FDS), flexor digitorum profundus (FDP) and flexor pollicis longus (FPL) tendons respectively. With simulated active flexion, the mean tendon excursion was 14 mm, 10 mm and 11 mm respectively. The mean tendon excursion in clinical cases intraoperatively following passive flexion was 2 mm for FDS, FDP and FPL respectively; following simulated active flexion it was 10 mm, 11 mm and 11 mm for FDS, FDP and FPL respectively. On the tenth day following repair, the mean excursions of FDS, FDP and FPL were 1 mm, 4 mm and 4 mm on passive flexion as compared to 3 mm, 10 mm and 12 mm on active flexion respectively. Three weeks postoperatively, the mean excursions of FDS, FDP and FPL tendons were 1 mm, 2 mm and 1 mm on passive flexion as compared to 5 mm, 15 mm on active flexion respectively. Six weeks postoperatively, the mean excursions of FDS, FDP and FPL tendons were 9 mm, 7 mm and 4 mm on passive flexion as compared to 12 mm, 33 mm and 20 mm on active flexion respectively. These results demonstrate an increased excursion of repaired flexor tendons in Zone V following an active flexion mobilisation regime as compared to a passive flexion mobilisation regime.

  7. The active translation of MHCII mRNA during dendritic cells maturation supplies new molecules to the cell surface pool.

    Malanga, Donatella; Barba, Pasquale; Harris, Paul E; Maffei, Antonella; Del Pozzo, Giovanna

    2007-04-01

    The transition of human dendritic cells (DCs) from the immature to the mature phenotype is characterized by an increased density of MHC class II (MHCII) molecules on the plasma membrane, a key requirement of their competence as professional antigen presenting cells (APCs). MHCII molecules on the cell surface derive from newly synthesized as well as from preexisting proteins. So far, all the studies done on DCs during maturation, to establish the relative contribution of newly synthesized MHCII molecules to the cell surface pool did not produced a clear, unified scenario. We report that, in human DCs stimulated ex vivo with LPS, the changes in the RNA accumulation specific for at least two MHCII genes (HLA-DRA and HLA-DQA1) due to transcriptional upregulation, is associated with the active translation at high rate of these transcripts. Our finding reveals that, across the 24h of the maturation process in human DCs, newly synthesized MHCII proteins are supplied to the APCs cell surface pool.

  8. Sequence analysis of morbillivirus CD150 receptor-Signaling Lymphocyte Activation Molecule (SLAM) of different animal species.

    Sarkar, J; Balamurugan, V; Sen, A; Saravanan, P; Sahay, B; Rajak, K K; Rasool, T J; Bhanuprakash, V; Singh, R K

    2009-12-01

    Signaling Lymphocyte Activation Molecule-SLAM (CD150) molecule has been reported as a putative receptor for most morbilliviruses for their respective host species. In this study, we determined the complete nucleotide sequence of the gene coding for the morbillivirus receptor-SLAM from the four species, namely, goat (Capra hircus), sheep (Ovis aries), Indian cattle (Bos indicus), and buffalo (Bubalus bubalis). The nucleotide (nt) open reading frame sequence of SLAM gene in all the four species studied was 1017 nucleotides in length encoding a polypeptide of 339 amino acids (aa), similar to Bos taurus, but different from canine, human, marmoset, and mouse SLAM, which were 1029, 1008, 1011, and 1032 nts, respectively, in length, and coding for 343, 336, 337, and 344 aa, respectively. Sequence analysis revealed 96.3-98.5% and 92.9-96.8% identities among the four species at the nt and aa level, respectively. Sequence diversity at aa level between various species revealed that the critical functional region of SLAM protein among different species is relatively conserved, thereby facilitating this molecule to act as a receptor for morbillivirus. Phylogenetic relationship based on the aa sequences of SLAM protein revealed that caprine, ovine, cattle, and buffalo fall under a defined cluster but caprine SLAM is more closely related to ovine, followed by bovine.

  9. Xalpha-DVM investigation of double water molecule interactions with active sites of alpha- and beta-subunits of hemoglobin

    Yuryeva, Elmira I.

    In this work, the results of Xalpha-discrete variation method calculations of the electronic structure and interatomic parameters of chemical bonding between iron (II) and oxygen molecule with and without extra electrons and protons in active site (AS) of alpha- and beta-subunits of oxyhemoglobin are presented. The Skulachev model of O2 molecule existing in respiration medium in the 2H2O form was used. The introduction of extra electrons does not change considerably the interaction of the iron atom with the O2 oxygen molecule, but strengthens the repulsion in the Fe bond N bonds. In this case, the estimated effective charge of the iron atom is +1.8/1.5e for AS of alpha-/beta-subunits of oxyhemoglobin, and the magnetic moment of iron atoms becomes zero. The deoxygenation effect of the AS of the alpha- and beta-subunits of oxyhemoglobin is due to the ability of extra protons to break down covalent attraction between the iron atom and the nearest oxygen atom and also to weakening of the repulsive component of the covalent Fe bond N interactions.

  10. SC1, an immunoglobulin-superfamily cell adhesion molecule, is involved in the brain metastatic activity of lung cancer cells

    KUBOTA, YUKA; KIRIMURA, NAOKI; SHIBA, HATSUKI; ADACHI, KAZUHIDE; TSUKAMOTO, YASUHIRO

    2015-01-01

    SC1 is a cell adhesion molecule that belongs to the immunoglobulin superfamily; this molecule was initially purified from the chick embryonic nervous system and was reported to exhibit homophilic adhesion activity. SC1 is transiently expressed in various organs during development and has been identified in numerous neoplastic tissues, including lung cancer and colorectal carcinomas. The present study focused on the encephalic metastasis of lung cancer cells with respect to the potential function of SC1, as this molecule is known to be consistently expressed in the central nervous system as well as lung cancers. SC1 complementary DNA was introduced into A549 cells, a human lung cancer-derived cell line. The stable overexpression of the SC1 protein in A549 cells was demonstrated to enhance the self-aggregation of the cells. In addition, the SC1 transfectants enhanced the metastatic and invasive potential to the encephalic parenchyma following implantation into nude mice. In conclusion, the results of the present study demonstrated that cell adhesion due interactions between SC1 on brain tissue and SC1 on lung cancer cells was involved in the malignant aspects of lung cancer, including invasion and metastasis to the brain. PMID:26622821

  11. Raman and surface enhanced Raman spectroscopic studies of specific, small molecule activator of histone acetyltransferase p300

    Kundu, Partha P.; Pavan Kumar, G. V.; Mantelingu, Kempegowda; Kundu, Tapas K.; Narayana, Chandrabhas

    2011-07-01

    We report for the first time, the Raman and surface enhanced Raman scattering (SERS) studies of N-(4-chloro-3-trifluoromethyl-phenyl)-2-ethoxy-benzamide (CTB). This molecule is specific activator of human histone acetyltransferase (HAT), p300, and serves as lead molecule to design anti-neoplastic therapeutics. A detailed Raman and SERS band assignments have been performed for CTB, which are compared with the density functional theory calculations. The observed red shift of N sbnd H stretching frequency from the computed wavenumber indicates the weakening of N sbnd H bond resulting from proton transfer to the neighboring oxygen atom. We observe Ag sbnd N vibrational mode at 234 cm -1 in SERS of CTB. This indicates there is a metal-molecule bond leading to chemical enhancement in SERS. We also observe, enhancement in the modes pertaining to substituted benzene rings and methyl groups. Based on SERS analysis we propose the adsorption sites and the orientation of CTB on silver surface.

  12. Conformational transition of FGFR kinase activation revealed by site-specific unnatural amino acid reporter and single molecule FRET

    Perdios, Louis; Lowe, Alan R.; Saladino, Giorgio; Bunney, Tom D.; Thiyagarajan, Nethaji; Alexandrov, Yuriy; Dunsby, Christopher; French, Paul M. W.; Chin, Jason W.; Gervasio, Francesco Luigi; Tate, Edward W.; Katan, Matilda

    2017-01-01

    Protein kinases share significant structural similarity; however, structural features alone are insufficient to explain their diverse functions. Thus, bridging the gap between static structure and function requires a more detailed understanding of their dynamic properties. For example, kinase activation may occur via a switch-like mechanism or by shifting a dynamic equilibrium between inactive and active states. Here, we utilize a combination of FRET and molecular dynamics (MD) simulations to probe the activation mechanism of the kinase domain of Fibroblast Growth Factor Receptor (FGFR). Using genetically-encoded, site-specific incorporation of unnatural amino acids in regions essential for activation, followed by specific labeling with fluorescent moieties, we generated a novel class of FRET-based reporter to monitor conformational differences corresponding to states sampled by non phosphorylated/inactive and phosphorylated/active forms of the kinase. Single molecule FRET analysis in vitro, combined with MD simulations, shows that for FGFR kinase, there are populations of inactive and active states separated by a high free energy barrier resulting in switch-like activation. Compared to recent studies, these findings support diversity in features of kinases that impact on their activation mechanisms. The properties of these FRET-based constructs will also allow further studies of kinase dynamics as well as applications in vivo.

  13. Conformational transition of FGFR kinase activation revealed by site-specific unnatural amino acid reporter and single molecule FRET

    Perdios, Louis; Lowe, Alan R.; Saladino, Giorgio; Bunney, Tom D.; Thiyagarajan, Nethaji; Alexandrov, Yuriy; Dunsby, Christopher; French, Paul M. W.; Chin, Jason W.; Gervasio, Francesco Luigi; Tate, Edward W.; Katan, Matilda

    2017-01-01

    Protein kinases share significant structural similarity; however, structural features alone are insufficient to explain their diverse functions. Thus, bridging the gap between static structure and function requires a more detailed understanding of their dynamic properties. For example, kinase activation may occur via a switch-like mechanism or by shifting a dynamic equilibrium between inactive and active states. Here, we utilize a combination of FRET and molecular dynamics (MD) simulations to probe the activation mechanism of the kinase domain of Fibroblast Growth Factor Receptor (FGFR). Using genetically-encoded, site-specific incorporation of unnatural amino acids in regions essential for activation, followed by specific labeling with fluorescent moieties, we generated a novel class of FRET-based reporter to monitor conformational differences corresponding to states sampled by non phosphorylated/inactive and phosphorylated/active forms of the kinase. Single molecule FRET analysis in vitro, combined with MD simulations, shows that for FGFR kinase, there are populations of inactive and active states separated by a high free energy barrier resulting in switch-like activation. Compared to recent studies, these findings support diversity in features of kinases that impact on their activation mechanisms. The properties of these FRET-based constructs will also allow further studies of kinase dynamics as well as applications in vivo. PMID:28045057

  14. Effect of methylprednisolone on the oxidative burst activity, adhesion molecules and clinical outcome following open heart surgery

    Toft, P; Christiansen, K; Tønnesen, Else Kirstine

    1997-01-01

    Following cardiac surgery with cardiopulmonary bypass (CPB), activated granulocytes may be involved with ischaemia/ reperfusion injury. The purpose of this study was to investigate whether steroids could reduce the oxidative burst activity of granulocytes, the expression of adhesion molecules...... on granulocytes and improve clinical outcome. Sixteen patients undergoing open heart surgery participated in the study. Eight were randomized to receive methylprednisolone (30 mg/kg intravenously) at the start of anaesthesia while eight patients served as a control group. The oxidative burst was measured flow...... not improve the weaning from the ventilator or reduce the stay in the intensive-care unit. In conclusion, treatment with steroids prevented hyperthermia following open heart surgery with CPB and reduced capillary leak during ECC. Methylprednisolone, however, did not reduce the oxidative burst activity...

  15. Effect of methylprednisolone on the oxidative burst activity, adhesion molecules and clinical outcome following open heart surgery

    Toft, P; Christiansen, K; Tønnesen, Else Kirstine;

    1997-01-01

    on granulocytes and improve clinical outcome. Sixteen patients undergoing open heart surgery participated in the study. Eight were randomized to receive methylprednisolone (30 mg/kg intravenously) at the start of anaesthesia while eight patients served as a control group. The oxidative burst was measured flow...... not improve the weaning from the ventilator or reduce the stay in the intensive-care unit. In conclusion, treatment with steroids prevented hyperthermia following open heart surgery with CPB and reduced capillary leak during ECC. Methylprednisolone, however, did not reduce the oxidative burst activity......Following cardiac surgery with cardiopulmonary bypass (CPB), activated granulocytes may be involved with ischaemia/ reperfusion injury. The purpose of this study was to investigate whether steroids could reduce the oxidative burst activity of granulocytes, the expression of adhesion molecules...

  16. An Intramolecular Silylene Borane Capable of Facile Activation of Small Molecules, Including Metal-Free Dehydrogenation of Water.

    Mo, Zhenbo; Szilvási, Tibor; Zhou, Yu-Peng; Yao, Shenglai; Driess, Matthias

    2017-02-27

    The first single-component N-heterocyclic silylene borane 1 (LSi-R-BMes2 ; L=PhC(N(t) Bu)2 ; R=1,12-xanthendiyl spacer; Mes=2,4,6-Me3 C6 H2 ), acting as a frustrated Lewis pair (FLP) in small-molecule activation, can be synthesized in 65 % yields. Its HOMO is largely localized at the silicon(II) atom and the LUMO has mainly boron 2p character. In small-molecule activation 1 allows access to the intramolecular silanone-borane 3 featuring a Si=O→B interaction through reaction with O2 , N2 O, or CO2 , and formation of silanethione borane 4 from reaction with S8 . The Si(II) center in 1 undergoes immediate hydrogenation if exposed to H2 at 1 atm pressure in benzene, affording the silane borane 5-H2 , L(H2 )Si-R-BMes2 . Remarkably, no H2 activation occurs if the single silylene LSiPh and Mes3 B intermolecularly separated are exposed to dihydrogen. Unexpectedly, the pre-organized Si-B separation in 1 enables a metal-free dehydrogenation of H2 O to give the silanone-borane 3 as reactive intermediate.

  17. PdM (M = Pt, Au) bimetallic alloy nanowires with enhanced electrocatalytic activity for electro-oxidation of small molecules

    Zhu, Chengzhou; Guo, Shaojun; Dong, Shaojun [State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin, 130022 (China)

    2012-05-02

    A facile and general method has been developed to synthesize well-defined PdPt and PdAu alloy nanowires, which exhibit significantly enhanced activity towards small molecules, such as ethanol, methanol, and glucose electro-oxidation in an alkaline medium. Considering the important role of one-dimensional alloy nanowires in electrocatalytic systems, the present Pd-based alloy nanostructures could offer a promising new class of advanced electrocatalysts for direct alcohol fuel cells and electrochemical sensors. (Copyright copyright 2012 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  18. The Salton Seismic Imaging Project (SSIP): Active Rift Processes in the Brawley Seismic Zone

    Han, L.; Hole, J. A.; Stock, J. M.; Fuis, G. S.; Rymer, M. J.; Driscoll, N. W.; Kent, G.; Harding, A. J.; Gonzalez-Fernandez, A.; Lazaro-Mancilla, O.

    2011-12-01

    The Salton Seismic Imaging Project (SSIP), funded by NSF and USGS, acquired seismic data in and across the Salton Trough in southern California and northern Mexico in March 2011. The project addresses both rifting processes at the northern end of the Gulf of California extensional province and earthquake hazards at the southern end of the San Andreas Fault system. Seven lines of onshore refraction and low-fold reflection data were acquired in the Coachella, Imperial, and Mexicali Valleys, two lines and a grid of airgun and OBS data were acquired in the Salton Sea, and onshore-offshore data were recorded. Almost 2800 land seismometers and 50 OBS's were used in almost 5000 deployments at almost 4300 sites, in spacing as dense as 100 m. These instruments received seismic signals from 126 explosive shots up to 1400 kg and over 2300 airgun shots. In the central Salton Trough, North American lithosphere appears to have been rifted completely apart. Based primarily on a 1979 seismic refraction project, the 20-22 km thick crust is apparently composed entirely of new crust added by magmatism from below and sedimentation from above. Active rifting of this new crust is manifested by shallow (geothermal energy production. This presentation is focused on an onshore-offshore line of densely sampled refraction and low-fold reflection data that crosses the Brawley Seismic Zone and Salton Buttes in the direction of plate motion. At the time of abstract submission, data analysis was very preliminary, consisting of first-arrival tomography of the onshore half of the line for upper crustal seismic velocity. Crystalline basement (>5 km/s), comprised of late-Pliocene to Quaternary sediment metamorphosed by the high heat flow, occurs at ~2 km depth beneath the Salton Buttes and geothermal field and ~4 km depth south of the BSZ. Preliminary result suggests that the velocity of basement is lower in the BSZ than to the south, which may result from fracturing. Basement velocity appears to be

  19. Activation of toll-like receptors and dendritic cells by a broad range of bacterial molecules

    Boele, L.C.L.; Bajramovic, J.J.; Vries, A.M.M.B.C. de; Voskamp-Visser, I.A.I.; Kaman, W.E.; Kleij, D. van der

    2009-01-01

    Activation of pattern recognition receptors such as Toll-like receptors (TLRs) by pathogens leads to activation and maturation of dendritic cells (DC), which orchestrate the development of the adaptive immune response. To create an overview of the effects of a broad range of pathogenic bacteria, the

  20. High quality, small molecule-activity datasets for kinase research [version 3; referees: 2 approved

    Rajan Sharma

    2016-10-01

    Full Text Available Kinases regulate cell growth, movement, and death. Deregulated kinase activity is a frequent cause of disease. The therapeutic potential of kinase inhibitors has led to large amounts of published structure activity relationship (SAR data. Bioactivity databases such as the Kinase Knowledgebase (KKB, WOMBAT, GOSTAR, and ChEMBL provide researchers with quantitative data characterizing the activity of compounds across many biological assays. The KKB, for example, contains over 1.8M kinase structure-activity data points reported in peer-reviewed journals and patents. In the spirit of fostering methods development and validation worldwide, we have extracted and have made available from the KKB 258K structure activity data points and 76K associated unique chemical structures across eight kinase targets. These data are freely available for download within this data note.

  1. Synthesis and anti-tumor activity evaluation of gallic acid-mangiferin hybrid molecule.

    Hu, Xiang-yu; Deng, Jia-gang; Wang, Lin; Yuan, Ye-fei

    2013-12-01

    To improve the anti-tumor effects of gallic acid and mangiferin, a gallic acid-mangiferin hybrid molecule (GAMA) was synthesized from gallic acid with mangiferin in the presence of ionic liquid ChC1(choline chloride)·2SnC12. Chemical and spectroscopic methods, such as (1)H and (13)C NMR spectroscopy, and HR-ESIMS were used for the structure identification of GA-MA. Using the cell counting kit-8 (CCK-8) assay, the in vitro anti-tumor effects were compared between GA-MA, gallic acid and mangiferin on human hepatoma HepG2, human nasopharyngeal carcinoma CNE, human lung cancer NCI-H460, human ovarian cancer SK-OV-3, and human cervical cancer Hela cells. The results showed that the half inhibitory concentration (IC50) of GA-MA on HepG2, CNE, NCI-H460, SK-OV-3, and Hela cells was significantly lower than that of gallic acid or mangiferin. This showed that GA-MA has a better in vitro anti-tumor effect than gallic acid and mangi-ferin.

  2. Identification of novel orosensory active molecules in cured vanilla Beans (Vanilla planifolia).

    Schwarz, Bernd; Hofmann, Thomas

    2009-05-13

    Sequential application of solvent extraction, gel permeation chromatography, and HPLC in combination with taste dilution analyses, followed by LC-MS and 1D/2D NMR experiments, led to the discovery of seven velvety mouth-coating molecules in cured beans of Vanilla planifolia . Among these, 5-(4-hydroxybenzyl)vanillin, 4-(4-hydroxybenzyl)-2-methoxyphenol, 4-hydroxy-3-(4-hydroxy-3-methoxybenzyl)-5-methoxybenzaldehyde, (1-O-vanilloyl)-(6-O-feruloyl)-beta-d-glucopyranoside, americanin A, and 4',6'-dihydroxy-3',5-dimethoxy-[1,1'-biphenyl]-3-carboxaldehyde were previously not reported in vanilla beans. Sensory studies revealed human recognition thresholds for the velvety mouth-coating sensation between 1.0 and 5.0 mumol/kg (water). Interestingly, the biphenyl derivatives were found to enhance the perception of creaminess and fatty body of sweetened skim milk, among which 4',6'-dihydroxy-3',5-dimethoxy-[1,1'-biphenyl]-3-carboxaldehyde showed the lowest threshold level of 5 mumol/kg. Quantitative analysis of these compounds in cured vanilla beans from different origins as well as in noncured beans revealed that, with the exception of americanin A, all of the other taste compounds are not present in the green vanilla beans and are formed during the bean curing process.

  3. Single Molecule Instrument for Surface Enhanced Raman Optical Activity of Biomolecules Project

    National Aeronautics and Space Administration — Stereochemistry is an essential element of our organic life. Only certain enantiomers are useful as drugs for the human body. Raman Optical Activity (ROA) and...

  4. Single Molecule Instrument for Surface Enhanced Raman Optical Activity of Biomolecules Project

    National Aeronautics and Space Administration — Stereochemistry is an essential element of our organic life. Only certain enantiomers are useful as drugs for the human body. Raman optical activity (ROA) provides...

  5. Evaluation of approximate measurements of activation-free-energy spectra of shear transformation zones in metallic glasses

    Ju, JongDoo [Department of Nuclear Engineering and Radiological Sciences, The University of Michigan, Ann Arbor, MI (United States); Atzmon, Michael, E-mail: atzmon@umich.edu [Department of Nuclear Engineering and Radiological Sciences, The University of Michigan, Ann Arbor, MI (United States); Department of Materials Science and Engineering, The University of Michigan, Ann Arbor, MI (United States)

    2015-09-15

    Highlights: • We simulate measured activation energy spectra for anelastic relaxation in metallic glasses. • Simulated spectra exhibit the same shape as earlier data obtained by temperature stepping. • The drop previously observed at high activation free energy is an artifact of the measurements. - Abstract: For many years, the only experimental activation free energy spectrum for shear transformations in metallic glasses had been obtained by quenching from high temperature during creep, followed by temperature stepping (Argon and Kuo, 1980). We show that the approximation associated with attributing a single activation energy to each temperature leads to an artificial drop in the spectrum at high activation energies. The detailed spectra of potential shear transformation zones we have recently obtained, which exhibit an atomically quantized hierarchy and are monotonic, lead to the same spectrum shape as obtained by Argon and Kuo when the approximation inherent to the temperature stepping method is applied.

  6. Isolation and molecular characterization of biofouling bacteria and profiling of quorum sensing signal molecules from membrane bioreactor activated sludge.

    Lade, Harshad; Paul, Diby; Kweon, Ji Hyang

    2014-02-04

    The formation of biofilm in a membrane bioreactor depends on the production of various signaling molecules like N-acyl homoserine lactones (AHLs). In the present study, a total of 200 bacterial strains were isolated from membrane bioreactor activated sludge and screened for AHLs production using two biosensor systems, Chromobacterium violaceum CV026 and Agrobacterium tumefaciens A136. A correlation between AHLs production and biofilm formation has been made among screened AHLs producing strains. The 16S rRNA gene sequence analysis revealed the dominance of Aeromonas and Enterobacter sp. in AHLs production; however few a species of Serratia, Leclercia, Pseudomonas, Klebsiella, Raoultella and Citrobacter were also identified. The chromatographic characterization of sludge extract showed the presence of a broad range of quorum sensing signal molecules. Further identification of sludge AHLs by thin layer chromatography bioassay and high performance liquid chromatography confirms the presence of C4-HSL, C6-HSL, C8-HSL, 3-oxo-C8-HSL, C10-HSL, C12-HSL, 3-oxo-C12-HSL and C14-HSL. The occurrence of AHLs in sludge extract and dominance of Aeromonas and Enterobacter sp. in activated sludge suggests the key role of these bacterial strains in AHLs production and thereby membrane fouling.

  7. Isolation and Molecular Characterization of Biofouling Bacteria and Profiling of Quorum Sensing Signal Molecules from Membrane Bioreactor Activated Sludge

    Harshad Lade

    2014-02-01

    Full Text Available The formation of biofilm in a membrane bioreactor depends on the production of various signaling molecules like N-acyl homoserine lactones (AHLs. In the present study, a total of 200 bacterial strains were isolated from membrane bioreactor activated sludge and screened for AHLs production using two biosensor systems, Chromobacterium violaceum CV026 and Agrobacterium tumefaciens A136. A correlation between AHLs production and biofilm formation has been made among screened AHLs producing strains. The 16S rRNA gene sequence analysis revealed the dominance of Aeromonas and Enterobacter sp. in AHLs production; however few a species of Serratia, Leclercia, Pseudomonas, Klebsiella, Raoultella and Citrobacter were also identified. The chromatographic characterization of sludge extract showed the presence of a broad range of quorum sensing signal molecules. Further identification of sludge AHLs by thin layer chromatography bioassay and high performance liquid chromatography confirms the presence of C4-HSL, C6-HSL, C8-HSL, 3-oxo-C8-HSL, C10-HSL, C12-HSL, 3-oxo-C12-HSL and C14-HSL. The occurrence of AHLs in sludge extract and dominance of Aeromonas and Enterobacter sp. in activated sludge suggests the key role of these bacterial strains in AHLs production and thereby membrane fouling.

  8. Identification of the phospholipid lysobisphosphatidic acid in the protozoan Entamoeba histolytica: An active molecule in endocytosis

    Silvia Castellanos-Castro

    2016-03-01

    Full Text Available Phospholipids are essential for vesicle fusion and fission and both are fundamental events for Entamoeba histolytica phagocytosis. Our aim was to identify the lysobisphosphatidic acid (LBPA in trophozoites and investigate its cellular fate during endocytosis. LBPA was detected by TLC in a 0.5 Rf spot of total lipids, which co-migrated with the LBPA standard. The 6C4 antibody, against LBPA recognized phospholipids extracted from this spot. Reverse phase LC-ESI-MS and MS/MS mass spectrometry revealed six LBPA species of m/z 772.58–802.68. LBPA was associated to pinosomes and phagosomes. Intriguingly, during pinocytosis, whole cell fluorescence quantification showed that LBPA dropped 84% after 15 min incubation with FITC-Dextran, and after 60 min, it increased at levels close to steady state conditions. Similarly, during erythrophagocytosis, after 15 min, LBPA also dropped in 36% and increased after 60 and 90 min. EhRab7A protein appeared in some vesicles with LBPA in steady state conditions, but after phagocytosis co-localization of both molecules increased and in late phases of erythrophagocytosis they were found in huge phagosomes or multivesicular bodies with many intraluminal vacuoles, and surrounding ingested erythrocytes and phagosomes. The 6C4 and anti-EhADH (EhADH is an ALIX family protein antibodies and Lysotracker merged in about 50% of the vesicles in steady state conditions and throughout phagocytosis. LBPA and EhADH were also inside huge phagosomes. These results demonstrated that E. histolytica LBPA is associated to pinosomes and phagosomes during endocytosis and suggested differences of LBPA requirements during pinocytosis and phagocytosis.

  9. Phytohormones as Important Biologically Active Molecules – Their Simple Simultaneous Detection

    Ladislav Havel

    2009-05-01

    Full Text Available Phytohormones, their functions, synthesis and effects, are of great interest. To study them in plant tissues accurate and sensitive methods are required. In the present study we aimed at optimizing experimental conditions to separate and determine not only plant hormones but also their metabolites, by liquid chromatography coupled with a UV-VIS detector. The mixture we analyzed was composed of benzyladenine, kinetin, trans-zeatin, cis-zeatin, dihydrozeatin, meta-topolin, ortho-topolin, α-naphthalene acetic acid, indole-3-acetic acid, trans-zeatin-7-glucoside, trans-zeatin-O-glucoside, trans-zeatin-9-riboside, meta-topolin-9-riboside and ortho-topolin-9-riboside. We measured the calibration dependences and estimated limits of detection and quantification under the optimal chromatographic conditions (column: Polaris C18; mobile phase: gradient starting at 2:98 (methanol:0.001% TFA and was increasing to 55:45 during twenty minutes, and then decreasing for 10 min to 35:65, flow rate: 200 µL·min-1, temperature: 50 °C, wavelength: 210 nm. The detection limits for the target molecules were estimated as tens of ng per mL. We also studied the effect of flax extracts on the phytohormones’ signals. Recovery of aliphatic and aromatic cytokinins, metabolites of cytokinins and auxinswere within the range from 87 to 105 %. The experimental conditions were tested on a mass selective detector. In addition we analysed a commercial product used for stimulation of roots formation in cuttings of poorly rooting plants. The determined content of α-naphthalene acetic acid was in good agreement with that declared by the manufacturer.

  10. Small Molecule APY606 Displays Extensive Antitumor Activity in Pancreatic Cancer via Impairing Ras-MAPK Signaling

    Guo, Na; Liu, Zuojia; Zhao, Wenjing; Wang, Erkang; Wang, Jin

    2016-01-01

    Pancreatic cancer has been found with abnormal expression or mutation in Ras proteins. Oncogenic Ras activation exploits their extensive signaling reach to affect multiple cellular processes, in which the mitogen-activated protein kinase (MAPK) signaling exerts important roles in tumorigenesis. Therapies targeted Ras are thus of major benefit for pancreatic cancer. Although small molecule APY606 has been successfully picked out by virtual drug screening based on Ras target receptor, its in-depth mechanism remains to be elucidated. We herein assessed the antitumor activity of APY606 against human pancreatic cancer Capan-1 and SW1990 cell lines and explored the effect of Ras-MAPK and apoptosis-related signaling pathway on the activity of APY606. APY606 treatment resulted in a dose- and time-dependent inhibition of cancer cell viability. Additionally, APY606 exhibited strong antitumor activity, as evidenced not only by reduction in tumor cell invasion, migration and mitochondrial membrane potential but also by alteration in several apoptotic indexes. Furthermore, APY606 treatment directly inhibited Ras-GTP and the downstream activation of MAPK, which resulted in the down-regulation of anti-apoptotic protein Bcl-2, leading to the up-regulation of mitochondrial apoptosis pathway-related proteins (Bax, cytosolic Cytochrome c and Caspase 3) and of cyclin-dependent kinase 2 and Cyclin A, E. These data suggest that impairing Ras-MAPK signaling is a novel mechanism of action for APY606 during therapeutic intervention in pancreatic cancer. PMID:27223122

  11. Modulation of Pantothenate Kinase 3 Activity by Small Molecules that Interact with the Substrate/Allosteric Regulatory Domain

    Leonardi, Roberta; Zhang, Yong-Mei; Yun, Mi-Kyung; Zhou, Ruobing; Zeng, Fu-Yue; Lin, Wenwei; Cui, Jimmy; Chen, Taosheng; Rock, Charles O.; White, Stephen W.; Jackowski, Suzanne (SJCH)

    2010-09-27

    Pantothenate kinase (PanK) catalyzes the rate-controlling step in coenzyme A (CoA) biosynthesis. PanK3 is stringently regulated by acetyl-CoA and uses an ordered kinetic mechanism with ATP as the leading substrate. Biochemical analysis of site-directed mutants indicates that pantothenate binds in a tunnel adjacent to the active site that is occupied by the pantothenate moiety of the acetyl-CoA regulator in the PanK3 acetyl-CoA binary complex. A high-throughput screen for PanK3 inhibitors and activators was applied to a bioactive compound library. Thiazolidinediones, sulfonylureas and steroids were inhibitors, and fatty acyl-amides and tamoxifen were activators. The PanK3 activators and inhibitors either stimulated or repressed CoA biosynthesis in HepG2/C3A cells. The flexible allosteric acetyl-CoA regulatory domain of PanK3 also binds the substrates, pantothenate and pantetheine, and small molecule inhibitors and activators to modulate PanK3 activity.

  12. Active source monitoring at the Wenchuan fault zone: coseismic velocity change associated with aftershock event and its implication

    Yang, Wei; Ge, Hongkui; Wang, Baoshan; Hu, Jiupeng; Yuan, Songyong; Qiao, Sen

    2014-12-01

    With the improvement of seismic observation system, more and more observations indicate that earthquakes may cause seismic velocity change. However, the amplitude and spatial distribution of the velocity variation remains a controversial issue. Recent active source monitoring carried out adjacent to Wenchuan Fault Scientific Drilling (WFSD) revealed unambiguous coseismic velocity change associated with a local M s5.5 earthquake. Here, we carry out forward modeling using two-dimensional spectral element method to further investigate the amplitude and spatial distribution of observed velocity change. The model is well constrained by results from seismic reflection and WFSD coring. Our model strongly suggests that the observed coseismic velocity change is localized within the fault zone with width of ~120 m rather than dynamic strong ground shaking. And a velocity decrease of ~2.0 % within the fault zone is required to fit the observed travel time delay distribution, which coincides with rock mechanical experiment and theoretical modeling.

  13. Holocene activities of the Taigu fault zone,Shanxi Province, and their relations with the 1303 Hongdong M=8 earthquake

    谢新生; 江娃利; 王焕贞; 冯西英

    2004-01-01

    The Taigu fault zone is one of the major 12 active boundary faults of the Shanxi fault-depression system, locatedon the eastern boundary of the Jinzhong basin. As the latest investigation indicated, the fault zone had dislocatedgully terrace of the f1rst order, forming fault-scarp in front of the loess mesa. It has been discovered in many placesin ground surface and trenches that Holocene deposits were dislocated. The latest activity was the 1303 Hongdongearthquake M=8, the fault appeared as right-lateral strike-slip with normal faulting. During that earthquake, theTaigu fault together with the Mianshan western-side fault on the Lingshi upheaval and the Huoshan pediment faulton the eastern boundary of the Linfen basin was being active, forming a surface rupture belt of 160 km in length.Moreover, the Taigu fault were active in the mid-stage of Holocene and near 7 700 aB.P. From these we learnt that,in Shanxi fault-depression system, the run-through activity of two boundary faults of depression-basins mightgenerate great earthquake with M=8.

  14. Current parallel chemistry principles and practice: application to the discovery of biologically active molecules.

    Edwards, Paul J

    2009-11-01

    This article describes the use of parallel chemistry techniques for drug discovery, based on publications from January 2006 to December 2008. Chemical libraries that yielded active compounds across a range of biological targets are presented, together with synthetic details when appropriate. Background information for the biological targets involved and any SAR that could be discerned within members of a library series also is discussed. New technological developments, as applied to library design and synthesis and, more generally, in the discovery of biologically active entities, are highlighted. In addition, the likely future directions for parallel chemistry in its ability to impact upon drug discovery are also presented.

  15. Cellular Activity of New Small Molecule Protein Arginine Deiminase 3 (PAD3) Inhibitors.

    Jamali, Haya; Khan, Hasan A; Tjin, Caroline C; Ellman, Jonathan A

    2016-09-08

    The protein arginine deiminases (PADs) catalyze the post-translational deimination of arginine side chains. Multiple PAD isozymes have been characterized, and abnormal PAD activity has been associated with several human disease states. PAD3 has been characterized as a modulator of cell growth via apoptosis inducing factor and has been implicated in the neurodegenerative response to spinal cord injury. Here, we describe the design, synthesis, and evaluation of conformationally constrained versions of the potent and selective PAD3 inhibitor 2. The cell activity of representative inhibitors in this series was also demonstrated for the first time by rescue of thapsigargin-induced cell death in PAD3-expressing HEK293T cells.

  16. TOPONYMS AS REFLECTION OF THE PECULIARITIES OF CRAFT ACTIVITY (ON THE MATERIAL OF SMOLENSK-VITEBSK BORDER-ZONE

    Evseeva Olga Sergeevna

    2014-09-01

    Full Text Available The geographical names are important units, representing the connection with the cultural and historical traditions and national-specific context of a specific time period; due to the inherent cumulative functions they fix and retain the essential features of a variety of information: linguistic, cultural, social, historical, psychological, ethnographic. The article deals with the toponyms, spread on the territory of the Smolensk-Vitebsk borderzone and recorded in modern lexicographical sources (toponymic and dialect dictionaries. The card index of geographical names collected by the author includes 1196 units, 604 of which are registered in the Smolensk border-zone and 592 in the Vitebsk border-zone. The conducted analysis allowed us to identify nine groups of the studied units, depending on the semantics of motivating word. Special attention is paid to the toponyms formed from appellatives associated with industrial activity, which was widely spread on the territory of Smolensk region in different historical periods. These units reflect the linguistic, historical, cultural and economic specifics of Smolensk-Vitebsk border-zone. The toponyms which present the development of mining, brick production, the manufacture of household utensils (glass and clay are the most frequent. The author reveals that the dialectal units and borrowed words confirming the close language contact among the frontier inhabitants, as well as Slavic vocabulary, were used as appellatives that served as the basis for toponyms. The results of the linguistic and culturological analysis allowed to conclude that the fragment of toponymic system, based on the craft activity of Smolensk-Vitebsk border-zone residents, testified to the community and the interaction between cultures and languages of closely related peoples – Russians and Belarusians.

  17. Dynamic fracturing by successive coseismic loadings leads to pulverization in active fault zones

    Aben, F. M.; Doan, M.-L.; Mitchell, T. M.; Toussaint, R.; Reuschlé, T.; Fondriest, M.; Gratier, J.-P.; Renard, F.

    2016-04-01

    Previous studies show that pulverized rocks observed along large faults can be created by single high-strain rate loadings in the laboratory, provided that the strain rate is higher than a certain pulverization threshold. Such loadings are analogous to large seismic events. In reality, pulverized rocks have been subject to numerous seismic events rather than one single event. Therefore, the effect of successive "milder" high-strain rate loadings on the pulverization threshold is investigated by applying loading conditions below the initial pulverization threshold. Single and successive loading experiments were performed on quartz-monzonite using a Split Hopkinson Pressure Bar apparatus. Damage-dependent petrophysical properties and elastic moduli were monitored by applying incremental strains. Furthermore, it is shown that the pulverization threshold can be reduced by successive "milder" dynamic loadings from strain rates of ~180 s-1 to ~90 s-1. To do so, it is imperative that the rock experiences dynamic fracturing during the successive loadings prior to pulverization. Combined with loading conditions during an earthquake rupture event, the following generalized fault damage zone structure perpendicular to the fault will develop: furthest from the fault plane, there is a stationary outer boundary that bounds a zone of dynamically fractured rocks. Closer to the fault, a pulverization boundary delimits a band of pulverized rock. Consecutive seismic events will cause progressive broadening of the band of pulverized rocks, eventually creating a wider damage zone observed in mature faults.

  18. Screening a Commercial Library of Pharmacologically Active Small Molecules against Staphylococcus aureus Biofilms.

    Torres, Nelson S; Abercrombie, Johnathan J; Srinivasan, Anand; Lopez-Ribot, Jose L; Ramasubramanian, Anand K; Leung, Kai P

    2016-10-01

    It is now well established that bacterial infections are often associated with biofilm phenotypes that demonstrate increased resistance to common antimicrobials. Further, due to the collective attrition of new antibiotic development programs by the pharmaceutical industries, drug repurposing is an attractive alternative. In this work, we screened 1,280 existing commercially available drugs in the Prestwick Chemical Library, some with previously unknown antimicrobial activity, against Staphylococcus aureus, one of the commonly encountered causative pathogens of burn and wound infections. From the primary screen of the entire Prestwick Chemical Library at a fixed concentration of 10 μM, 104 drugs were found to be effective against planktonic S. aureus strains, and not surprisingly, these were mostly antimicrobials and antiseptics. The activity of 18 selected repurposing candidates, that is, drugs that show antimicrobial activity that are not already considered antimicrobials, observed in the primary screen was confirmed in dose-response experiments. Finally, a subset of nine of these drug candidates was tested against preformed biofilms of S. aureus We found that three of these drugs, niclosamide, carmofur, and auranofin, possessed antimicrobial activity against preformed biofilms, making them attractive candidates for repurposing as novel antibiofilm therapies.

  19. Preconditioning of Carbon Monoxide Releasing Molecule-derived CO Attenuates LPS-induced Activation of HUVEC

    Bingwei Sun, Xiangqian Zou, Yueling Chen, Ping Zhang, Gengsheng Shi

    2008-01-01

    Full Text Available Objective: To investigate the effects and potential mechanisms of preconditioning of tricarbonyldichlororuthenium (III dimer (CORM-2-liberated CO on LPS-induced activation of endothelial cells (HUVEC. Methods: HUVEC were pretreated with CORM-2 at the concentration of 50 or 100μM for 2 hrs, washed and stimulated with LPS (10μg/ml for additional 4 hrs. Activation (oxidative stress of HUVEC was assessed by measuring intracellular oxidation of DHR 123 or nitration of DAF-FM, specific H2O2 and NO fluorochromes, respectively. The expression of HO-1, iNOS (Western blot and ICAM-1 (cell ELISA proteins and activation of inflammation-relevant transcription factor, NF-κB (EMSA were assessed. In addition, PMN adhesion to HUVEC was also assessed. Results: The obtained data indicate that pretreatment of HUVEC with CORM-2 results in: 1 decrease of LPS-induced production of ROS and NO; 2 up-regulation of HO-1 but decrease in iNOS at the protein levels; 3 inhibition of LPS-induced activation of NF-κB; and 4 downregulation of expression of ICAM-1, and this was accompanied by a decrease of PMN adhesion to LPS-stimulated HUVEC. Conclusions: Preconditioning of CO liberated by CORM-2 elicited its anti-inflammatory effects by interfering with the induction of intracellular oxidative stress. In addition, it also supports the notion that CO is a potent inhibitor of iNOS and NF-κB.

  20. Interleukin 3 stimulates proliferation and triggers endothelial-leukocyte adhesion molecule 1 gene activation of human endothelial cells.

    Brizzi, M F; Garbarino, G; Rossi, P R; Pagliardi, G L; Arduino, C; Avanzi, G C; Pegoraro, L

    1993-06-01

    Proliferation and functional activation of endothelial cells within a tissue site of inflammation are regulated by humoral factors released by cells, such as T lymphocytes and monocytes, infiltrating the perivascular space. In the present study we investigated the effects of interleukin 3 (IL-3), an activated T lymphocyte-derived cytokine, on cultured human umbilical vein endothelial cells (HUVEC). Proliferative activity, evaluated both by estimation of the fraction of cells in the S phase and by direct cell count demonstrated that IL-3, at the dose of 25 ng/ml, enhances more than threefold both DNA synthesis and cell proliferation above baseline control conditions. Binding studies with radioiodinated ligand demonstrated that HUVEC constitutively express a smaller number of IL-3 binding sites (approximately 99 binding sites per cell, with an apparent Kd of 149 pM). Accordingly, molecular analysis showed the presence of transcripts for both alpha and beta subunits of the IL-3 receptor. Functional activation of endothelial cells was evaluated by the expression of the endothelial-leukocyte adhesion molecule 1 (ELAM-1) transcript and by leukocyte adhesion. The ELAM-1 gene transcript was clearly detectable 4 h after IL-3 addition and started to decrease after 12 h. Moreover, IL-3-induced ELAM-1 transcription was followed by enhanced adhesion of neutrophils and CD4+ T cells to HUVEC. The findings that IL-3 can stimulate both proliferation and functional activation of endothelial cells suggest that this cytokine can be involved in sustaining the process of chronic inflammation.

  1. IgE and Drug Allergy: Antibody Recognition of ‘Small’ Molecules of Widely Varying Structures and Activities

    Brian A. Baldo

    2014-01-01

    Full Text Available The variety of chemically diverse pharmacologically-active compounds administered to patients is large and seemingly forever growing, and, with every new drug released and administered, there is always the potential of an allergic reaction. The most commonly occurring allergic responses to drugs are the type I, or immediate hypersensitivity reactions mediated by IgE antibodies. These reactions may affect a single organ, such as the nasopharynx (allergic rhinitis, eyes (conjunctivitis, mucosa of mouth/throat/tongue (angioedema, bronchopulmonary tissue (asthma, gastrointestinal tract (gastroenteritis and skin (urticaria, eczema, or multiple organs (anaphylaxis, causing symptoms ranging from minor itching and inflammation to death. It seems that almost every drug is capable of causing an immediate reaction and it is unusual to find a drug that has not provoked an anaphylactic response in at least one patient. These facts alone indicate the extraordinary breadth of recognition of IgE antibodies for drugs ranging from relatively simple structures, for example, aspirin, to complex molecules, such as the macrolide antibiotics composed of a large macrocyclic ring with attached deoxy sugars. This wide recognition profile is borne out at the molecular level by results of quantitative immunochemical studies where hapten inhibition investigations have identified structural determinants complementary to IgE antibodies in the sera of allergic subjects. Allergenic determinants have been identified on a variety of drugs including neuromuscular blockers, penicillins, cephalosporins, opioids, thiopentone, sulfonamides, trimethoprim, quinolones, chlorhexidine and the non-steroidal anti-inflammatory drug aspirin. It is already clear that IgE can distinguish fine structural differences on a wide variety of molecules, determinants may be at least as small as an amino group or encompass the whole molecule, and individual drugs may demonstrate allergenic heterogeneity.

  2. Diversity and dynamics of active microbial eukaryotes in the anoxic zone of a freshwater meromictic lake (Pavin, France

    CECILE eLEPERE

    2016-02-01

    Full Text Available Microbial eukaryotes play a crucial role in ecosystem functioning and oxygen is considered to be one of the strongest barriers against their local dispersal. However, diversity of microbial eukaryotes in freshwater habitats with oxygen gradients has previously received very little attention. We applied high-throughput sequencing (V4 region of the 18S rRNA gene in conjunction with quantitative PCR (DNA and RNA and fluorescent in situ hybridization analyses, to provide an unique spatio-temporal analysis of microbial eukaryotes diversity and potential activity in a meromictic freshwater lake (lake Pavin. This study revealed a high genetic diversity of unicellular eukaryotes in the permanent anoxic zone of lake Pavin and allowed the discrimination of active vs. inactive components. 42 % of the OTUs (Operational taxonomic Units are exclusively present in the monimolimnion, where Alveolata (Ciliophora and Dinophyceae and Fungi (Dikarya and Chytrids are the most active phyla and are probably represented by species capable of anaerobic metabolism. Pigmented eukaryotes (Haptophyceae and Chlorophyceae are also present and active in this zone, which opens up questions regarding their metabolism.

  3. Monolayer Formation Characteristics of Novel Organic Molecules with Nonlinear Optically Active Moieties

    1989-05-31

    reviews and special issues5 .6 . We recently reported7 the design and synthesis of a class of organic compounds ( Schiff base ) possessing a large second...RESULTS AND DISCUSSION The molecular structures of the candidate materials are shown in Fig. lb. The molecular structures of the NLO active Schiff base compounds...numerals, l-VII!. Step I in Fig. la shows the synthetic route to the Schiff base compounds MI-M6. Steps 2 and 3 define the approach to the synthesis of

  4. Genome-wide signatures of transcription factor activity: connecting transcription factors, disease, and small molecules.

    Jing Chen

    Full Text Available Identifying transcription factors (TF involved in producing a genome-wide transcriptional profile is an essential step in building mechanistic model that can explain observed gene expression data. We developed a statistical framework for constructing genome-wide signatures of TF activity, and for using such signatures in the analysis of gene expression data produced by complex transcriptional regulatory programs. Our framework integrates ChIP-seq data and appropriately matched gene expression profiles to identify True REGulatory (TREG TF-gene interactions. It provides genome-wide quantification of the likelihood of regulatory TF-gene interaction that can be used to either identify regulated genes, or as genome-wide signature of TF activity. To effectively use ChIP-seq data, we introduce a novel statistical model that integrates information from all binding "peaks" within 2 Mb window around a gene's transcription start site (TSS, and provides gene-level binding scores and probabilities of regulatory interaction. In the second step we integrate these binding scores and regulatory probabilities with gene expression data to assess the likelihood of True REGulatory (TREG TF-gene interactions. We demonstrate the advantages of TREG framework in identifying genes regulated by two TFs with widely different distribution of functional binding events (ERα and E2f1. We also show that TREG signatures of TF activity vastly improve our ability to detect involvement of ERα in producing complex diseases-related transcriptional profiles. Through a large study of disease-related transcriptional signatures and transcriptional signatures of drug activity, we demonstrate that increase in statistical power associated with the use of TREG signatures makes the crucial difference in identifying key targets for treatment, and drugs to use for treatment. All methods are implemented in an open-source R package treg. The package also contains all data used in the analysis

  5. Modelling active sites for the Beckmann rearrangement reaction in boron-containing zeolites and their interaction with probe molecules.

    Lezcano-González, Inés; Vidal-Moya, Alejandro; Boronat, Mercedes; Blasco, Teresa; Corma, Avelino

    2010-06-28

    Theoretical calculations and in situ solid state NMR spectroscopy have been combined to get insight on the nature of the active sites for the Beckmann rearrangement reaction in borosilicate zeolites. The interaction of a B site in zeolite Beta with a series of probe molecules (ammonia, pyridine, acetone and water) has been modelled and the (15)N and (11)B NMR isotropic chemical shift of the resulting complexes calculated and compared with experimental in situ NMR results. This approach has allowed validation of the methodology to model the adsorption on a zeolite boron site of molecules of varying basicity which are either protonated or non-protonated. The limitation is that theoretical calculations overestimate the effect of molecular adsorption through hydrogen bonds on the calculated isotropic (11)B NMR chemical shift.Theoretical and experimental results on the adsorption of acetophenone and cyclohexanone oximes on zeolite B-Beta indicate that Brønsted acid sites protonate the oximes, changing the boron coordination from trigonal to tetrahedral. Comparison of theoretical and experimental (15)N NMR chemical shifts of the adsorbed amides (acetanilide and epsilon-caprolactam) indicates that they are non-protonated, and the (11)B NMR spectra show that, as expected, boron remains in trigonal coordination with an isotropic delta(11)B(exp) which differs from the calculated value delta(11)B(calc).

  6. Enzyme kinetics and distinct modulation of the protein kinase N family of kinases by lipid activators and small molecule inhibitors

    Falk, Matthew D.; Liu, Wei; Bolaños, Ben; Unsal-Kacmaz, Keziban; Klippel, Anke; Grant, Stephan; Brooun, Alexei; Timofeevski, Sergei

    2014-01-01

    The PKN (protein kinase N) family of Ser/Thr protein kinases regulates a diverse set of cellular functions, such as cell migration and cytoskeletal organization. Inhibition of tumour PKN activity has been explored as an oncology therapeutic approach, with a PKN3-targeted RNAi (RNA interference)-derived therapeutic agent in Phase I clinical trials. To better understand this important family of kinases, we performed detailed enzymatic characterization, determining the kinetic mechanism and lipid sensitivity of each PKN isoform using full-length enzymes and synthetic peptide substrate. Steady-state kinetic analysis revealed that PKN1–3 follows a sequential ordered Bi–Bi kinetic mechanism, where peptide substrate binding is preceded by ATP binding. This kinetic mechanism was confirmed by additional kinetic studies for product inhibition and affinity of small molecule inhibitors. The known lipid effector, arachidonic acid, increased the catalytic efficiency of each isoform, mainly through an increase in kcat for PKN1 and PKN2, and a decrease in peptide KM for PKN3. In addition, a number of PKN inhibitors with various degrees of isoform selectivity, including potent (Ki<10 nM) and selective PKN3 inhibitors, were identified by testing commercial libraries of small molecule kinase inhibitors. This study provides a kinetic framework and useful chemical probes for understanding PKN biology and the discovery of isoform-selective PKN-targeted inhibitors. PMID:27919031

  7. Uncovering Single-Molecule Photophysical Heterogeneity of Bright, Thermally Activated Delayed Fluorescence Emitters Dispersed in Glassy Hosts.

    Noriega, Rodrigo; Barnard, Edward S; Ursprung, Benedikt; Cotts, Benjamin L; Penwell, Samuel B; Schuck, P James; Ginsberg, Naomi S

    2016-10-04

    Recently developed all-organic emitters used in display applications achieve high brightness by harvesting triplet populations via thermally activated delayed fluorescence. The photophysical properties of these emitters therefore involve new inherent complexities and are strongly affected by interactions with their host material in the solid state. Ensemble measurements occlude the molecular details of how host-guest interactions determine fundamental properties such as the essential balance of singlet oscillator strength and triplet harvesting. Therefore, using time-resolved fluorescence spectroscopy, we interrogate these emitters at the single-molecule level and compare their properties in two distinct glassy polymer hosts. We find that nonbonding interactions with aromatic moieties in the host appear to mediate the molecular configurations of the emitters, but also promote nonradiative quenching pathways. We also find substantial heterogeneity in the time-resolved photoluminescence of these emitters, which is dominated by static disorder in the polymer. Finally, since singlet-triplet cycling underpins the mechanism for increased brightness, we present the first room-temperature measurement of singlet-triplet equilibration dynamics in this family of emitters. Our observations present a molecular-scale interrogation of host-guest interactions in a disordered film, with implications for highly efficient organic light-emitting devices. Combining a single-molecule experimental technique with an emitter that is sensitive to triplet dynamics, yet read out via fluorescence, should also provide a complementary approach to performing fundamental studies of glassy materials over a large dynamic range of time scales.

  8. Human L-ficolin, a recognition molecule of the lectin activation pathway of complement, activates complement by binding to pneumolysin, the major toxin of Streptococcus pneumoniae.

    Ali, Youssif M; Kenawy, Hany I; Muhammad, Adnan; Sim, Robert B; Andrew, Peter W; Schwaeble, Wilhelm J

    2013-01-01

    The complement system is an essential component of the immune response, providing a critical line of defense against different pathogens including S. pneumoniae. Complement is activated via three distinct pathways: the classical (CP), the alternative (AP) and the lectin pathway (LP). The role of Pneumolysin (PLY), a bacterial toxin released by S. pneumoniae, in triggering complement activation has been studied in vitro. Our results demonstrate that in both human and mouse sera complement was activated via the CP, initiated by direct binding of even non-specific IgM and IgG3 to PLY. Absence of CP activity in C1q(-/-) mouse serum completely abolished any C3 deposition. However, C1q depleted human serum strongly opsonized PLY through abundant deposition of C3 activation products, indicating that the LP may have a vital role in activating the human complement system on PLY. We identified that human L-ficolin is the critical LP recognition molecule that drives LP activation on PLY, while all of the murine LP recognition components fail to bind and activate complement on PLY. This work elucidates the detailed interactions between PLY and complement and shows for the first time a specific role of the LP in PLY-mediated complement activation in human serum.

  9. Single-molecule catalysis mapping quantifies site-specific activity and uncovers radial activity gradient on single 2D nanocrystals.

    Andoy, Nesha May; Zhou, Xiaochun; Choudhary, Eric; Shen, Hao; Liu, Guokun; Chen, Peng

    2013-02-06

    Shape-controlled metal nanocrystals are a new generation of nanoscale catalysts. Depending on their shapes, these nanocrystals exhibit various surface facets, and the assignments of their surface facets have routinely been used to rationalize or predict their catalytic activity in a variety of chemical transformations. Recently we discovered that for 1-dimensional (1D) nanocrystals (Au nanorods), the catalytic activity is not constant along the same side facets of single nanorods but rather differs significantly and further shows a gradient along its length, which we attributed to an underlying gradient of surface defect density resulting from their linear decay in growth rate during synthesis (Nat. Nanotechnol.2012, 7, 237-241). Here we report that this behavior also extends to 2D nanocrystals, even for a different catalytic reaction. By using super-resolution fluorescence microscopy to map out the locations of catalytic events within individual triangular and hexagonal Au nanoplates in correlation with scanning electron microscopy, we find that the catalytic activity within the flat {111} surface facet of a Au nanoplate exhibits a 2D radial gradient from the center toward the edges. We propose that this activity gradient results from a growth-dependent surface defect distribution. We also quantify the site-specific activity at different regions within a nanoplate: The corner regions have the highest activity, followed by the edge regions and then the flat surface facets. These discoveries highlight the spatial complexity of catalytic activity at the nanoscale as well as the interplay amid nanocrystal growth, morphology, and surface defects in determining nanocatalyst properties.

  10. Kidney Injury Molecule-1 Protects against Gα12 Activation and Tissue Damage in Renal Ischemia-Reperfusion Injury

    Ismail, Ola Z.; Zhang, Xizhong; Wei, Junjun; Haig, Aaron; Denker, Bradley M.; Suri, Rita S.; Sener, Alp; Gunaratnam, Lakshman

    2016-01-01

    Ischemic acute kidney injury is a serious untreatable condition. Activation of the G protein α12 (Gα12) subunit by reactive oxygen species is a major cause of tissue damage during renal ischemia-reperfusion injury. Kidney injury molecule-1 (KIM-1) is a transmembrane glycoprotein that is highly up-regulated during acute kidney injury, but the physiologic significance of this up-regulation is unclear. Here, we report for the first time that Kim-1 inhibits Gα12 activation and protects mice against renal ischemia-reperfusion injury. We reveal that Kim-1 physically interacts with and inhibits cellular Gα12 activation after inflammatory stimuli, including reactive oxygen species, by blocking GTP binding to Gα12. Compared with Kim-1+/+ mice, Kim-1−/− mice exhibited greater Gα12 and downstream Src activation both in primary tubular epithelial cells after in vitro stimulation with H2O2 and in whole kidneys after unilateral renal artery clamping. Finally, we show that Kim-1–deficient mice had more severe kidney dysfunction and tissue damage after bilateral renal artery clamping, compared with wild-type mice. Our results suggest that KIM-1 is an endogenous protective mechanism against renal ischemia-reperfusion injury through inhibition of Gα12. PMID:25759266

  11. Time-dependent generalized-active-space configuration-interaction approach to photoionization dynamics of atoms and molecules

    Bauch, Sebastian; Madsen, Lars Bojer

    2014-01-01

    We present a wave-function based method to solve the time-dependent many-electron Schr\\"odinger equation (TDSE) with special emphasis on strong-field ionization phenomena. The theory builds on the configuration-interaction (CI) approach supplemented by the generalized-active-space (GAS) concept from quantum chemistry. The latter allows for a controllable reduction in the number of configurations in the CI expansion by imposing restrictions on the active orbital space. The method is similar to the recently formulated time-dependent restricted-active-space (TD-RAS) CI method [D. Hochstuhl, and M. Bonitz, Phys. Rev. A 86, 053424 (2012)]. We present details of our implementation and address convergence properties with respect to the active spaces and the associated account of electron correlation in both ground state and excitation scenarios. We apply the TD-GASCI theory to strong-field ionization of polar diatomic molecules and illustrate how the method allows us to uncover a strong correlation-induced shift of ...

  12. Specific small-molecule activator of Aurora kinase A induces autophosphorylation in a cell-free system.

    Kishore, A Hari; Vedamurthy, B M; Mantelingu, K; Agrawal, Shipra; Reddy, B A Ashok; Roy, Siddhartha; Rangappa, K S; Kundu, Tapas K

    2008-02-28

    Aurora kinases are essential for chromosomal segregation and cell division and thereby important for maintaining the proper genomic integrity. There are three classes of aurora kinases in humans: A, B, and C. Aurora kinase A is frequently overexpressed in various cancers. The link of the overexpression and tumorigenesis is yet to be understood. By employing virtual screening, we have found that anacardic acid, a pentadecane aliphatic chain containing hydroxylcarboxylic acid, from cashew nut shell liquid could be docked in Aurora kinases A and B. Remarkably, we found that anacardic acid could potently activate the Aurora kinase A mediated phosphorylation of histone H3, but at a similar concentration the activity of aurora kinase B remained unaffected in vitro. Mechanistically, anacardic acid induces the structural changes and also the autophosphorylation of the aurora kinase A to enhance the enzyme activity. This data thus indicate anacardic acid as the first small-molecule activator of Aurora kinase, which could be highly useful for probing the function of hyperactive (overexpressed) Aurora kinase A.

  13. The genomes of the non-clearing-zone-forming and natural-rubber- degrading species Gordonia polyisoprenivorans and Gordonia westfalica harbor genes expressing Lcp activity in Streptomyces strains.

    Bröker, Daniel; Dietz, David; Arenskötter, Matthias; Steinbüchel, Alexander

    2008-04-01

    The latex-clearing protein (Lcp(K30)) from the rubber-degrading bacterium Streptomyces sp. strain K30 is involved in the cleavage of poly(cis-1,4-isoprene), yielding isoprenoid aldehydes and ketones. Lcp homologues have so far been detected in all investigated clearing-zone-forming rubber-degrading bacteria. Internal degenerated oligonucleotides derived from lcp genes of Streptomyces sp. strain K30 (lcp(K30)), Streptomyces coelicolor strain A3(2), and Nocardia farcinica strains IFM10152 and E1 were applied in PCR to investigate whether lcp homologues occur also in the non-clearing-zone-forming rubber-utilizing bacteria Gordonia polyisoprenivorans strains VH2 and Y2K, Gordonia alkanivorans strain 44187, and Gordonia westfalica strain Kb1, which grow adhesively on rubber. The 1,230- and 1,224-bp lcp-homologous genes from G. polyisoprenivorans strain VH2 (lcp(VH2)) and G. westfalica strain Kb1 (lcp(Kb1)) were obtained after screening genomic libraries by degenerated PCR amplification, and their translational products exhibited 50 and 52% amino acid identity, respectively, to Lcp(K30). Recombinant lcp(VH2) and lcp(Kb1) harboring cells of the non-rubber-degrading Streptomyces lividans strain TK23 were able to form clearing zones and aldehydes on latex overlay-agar plates, thus indicating that lcp(VH2) and lcp(Kb1) encode functionally active proteins. Analysis by gel permeation chromatography demonstrated lower polymer concentrations and molecular weights of the remaining polyisoprenoid molecules after incubation with these recombinant S. lividans strains. Reverse transcription-PCR analysis demonstrated that lcp(VH2) was transcribed in cells of G. polyisoprenivorans strain VH2 cultivated in the presence of poly(cis-1,4-isoprene) but not in the presence of sodium acetate. Anti-Lcp(K30) immunoglobulin Gs, which were raised in this study, were rather specific for Lcp(K30) and did not cross-react with Lcp(VH2) and Lcp(Kb1). A lcp(VH2) disruption mutant was still able to grow

  14. High activity in catalytic cracking of large molecules over micro-mesoporous silicoaluminophosphate with controlled morphology

    2010-01-01

    A novel micro-mesoporous silicoaluminophosphate(MUS-5) with controlled morphology has been first synthesized in a two-step route.The physical properties of the silicoaluminophosphate were characterized using XRD,SEM,TEM,nitrogen adsorption-desorption and NH3-TPD techniques.When the pH value of the solution system was varied in the range from 2.0 to 5.0,three different morphologies of silicoaluminophosphate including chain-like,flower-like and barrel-like morphology were obtained.Catalytic tests showed that the silicoaluminophosphate exhibited higher catalytic activity compared with the conventional microporous SAPO-5 under the same conditions for catalytic cracking of 1,3,5-triisopropylbenzene heavy aromatics.The remarkable catalytic reactivity was mainly attributed to the presence of the hierarchical porosity in the silicoaluminophosphate catalyst.

  15. A structure-activity relationship study of small-molecule inhibitors of GLI1-mediated transcription.

    Actis, Marcelo; Connelly, Michele C; Mayasundari, Anand; Punchihewa, Chandanamali; Fujii, Naoaki

    2011-01-01

    We have previously reported ketoprofen amide compounds as inhibitors of GLI1-mediated transcription, an essential down-stream element of the Hedgehog (Hh) pathway. These compounds inhibited Gli-luciferase reporter in C3H10T1/2 cells that were exogenously transfected with GLI1 and in Rh30 cells that endogenously overexpress GLI1. Here we have designed new derivatives of these compounds aiming to explore the structure-activation relationship (SAR). By replacing the ketone carbonyl group of the ketoprofen moiety with an ether, amide, sulfonamide, or sulfone, we found several new compounds that are equipotent to the ketoprofen amide compounds. Among them, sulfone 30 inhibited Gli-luciferase reporter in C3H10T1/2 cells that were exogenously transfected with GLI1 and in Rh30 cells that endogenously overexpress GLI1.

  16. Probing binding and cellular activity of pyrrolidinone and piperidinone small molecules targeting the urokinase receptor.

    Mani, Timmy; Liu, Degang; Zhou, Donghui; Li, Liwei; Knabe, William Eric; Wang, Fang; Oh, Kyungsoo; Meroueh, Samy O

    2013-12-01

    The urokinase receptor (uPAR) is a cell-surface protein that is part of an intricate web of transient and tight protein interactions that promote cancer cell invasion and metastasis. Here, we evaluate the binding and biological activity of a new class of pyrrolidinone and piperidinone compounds, along with derivatives of previously-identified pyrazole and propylamine compounds. Competition assays revealed that the compounds displace a fluorescently labeled peptide (AE147-FAM) with inhibition constant (Ki ) values ranging from 6 to 63 μM. Structure-based computational pharmacophore analysis followed by extensive explicit-solvent molecular dynamics (MD) simulations and free energy calculations suggested the pyrazole-based and piperidinone-based compounds adopt different binding modes, despite their similar two-dimensional structures. In cells, pyrazole-based compounds showed significant inhibition of breast adenocarcinoma (MDA-MB-231) and pancreatic ductal adenocarcinoma (PDAC) cell proliferation, but piperidinone-containing compounds exhibited no cytotoxicity even at concentrations of 100 μM. One pyrazole-based compound impaired MDA-MB-231 invasion, adhesion, and migration in a concentration-dependent manner, while the piperidinone inhibited only invasion. The pyrazole derivative inhibited matrix metalloprotease-9 (gelatinase) activity in a concentration-dependent manner, while the piperidinone showed no effect suggesting different mechanisms for inhibition of cell invasion. Signaling studies further highlighted these differences, showing that pyrazole compounds completely inhibited ERK phosphorylation and impaired HIF1α and NF-κB signaling, while pyrrolidinones and piperidinones had no effect. Annexin V staining suggested that the effect of the pyrazole-based compound on proliferation was due to cell killing through an apoptotic mechanism. The compounds identified represent valuable leads in the design of further derivatives with higher affinities and

  17. Late Quaternary Activity and Seismogenic Potential of the Gonave Microplate: Plantain Garden Strike-Slip Fault Zone of Eastern Jamaica

    Mann, P.; Prentice, C.; King, W.; Demets, C.; Wiggins-Grandison, M.; Benford, B.

    2008-12-01

    At the longitude of Jamaica, Caribbean (Carib)-North America (Noam) plate motion of 19 ± 2 mm/a is carried by two parallel, left-lateral strike-slip faults, the Oriente fault zone, immediately south of Cuba, and the Enriquillo-Plantain Garden fault zone (EPGFZ), which lies 100-150 km further south. It has been postulated that the lithosphere between these faults constitutes an independent Gonave microplate that has formed in response to the ongoing collision between the leading edge of Carib in Hispaniola and the Bahama carbonate platform. GPS measurements in Jamaica and Hispanola is supportive of the microplate hypothesis and indicates that roughly half of Carib-Noam plate motion (8-14 mm/a) is carried by the EPGFZ of southern Hispaniola and eastern Jamaica. This study applies geomorphic and paleoseismic methods as a direct test of the activity and amount of microplate motion carried on the Plantain Garden fault segment of eastern Hispaniola and how this motion is distributed across a large restraining bend that has formed the island of Jamaica since the late Miocene. The EPFZ curves gently to the northeast and forming a steep mountain front to the Blue Mountains restraining bend with elevations up to 2200 m. Geomorphic fault-related features along the mountain front fault zone include left-laterally deflected rivers and streams, but no small scale features indicative of Holocene activity. River and stream deflections range from 0.1 to 0.5 km. We identified and trenched the most active trace of the mountain front fault at the Morant River where the fault is characterized by a 1.5-m-wide sub-vertical fault zone juxtaposing sheared alluvium and fault Cretaceous basement rocks This section is overlain by a 6-m-thick fluvial terrace. Trenching in the unfaulted terrace immediately overlying the fault trace revealed radiocarbon and OSL ages ranging from 20 to 21 ka that are consistent with a prominent unfaulted alluvial fan along the projection of this fault 1.5 km to

  18. High-resolution water column survey to identify active sublacustrine hydrothermal discharge zones within Lake Rotomahana, North Island, New Zealand

    Walker, Sharon L.; de Ronde, Cornel E. J.; Fornari, Daniel; Tivey, Maurice A.; Stucker, Valerie K.

    2016-03-01

    Autonomous underwater vehicles were used to conduct a high-resolution water column survey of Lake Rotomahana using temperature, pH, turbidity, and oxidation-reduction potential (ORP) to identify active hydrothermal discharge zones within the lake. Five areas with active sublacustrine venting were identified: (1) the area of the historic Pink Terraces; (2) adjacent to the western shoreline subaerial "Steaming Cliffs," boiling springs and geyser; (3) along the northern shoreline to the east of the Pink Terrace site; (4) the newly discovered Patiti hydrothermal system along the south margin of the 1886 Tarawera eruption rift zone; and (5) a location in the east basin (northeast of Patiti Island). The Pink Terrace hydrothermal system was active prior to the 1886 eruption of Mount Tarawera, but venting along the western shoreline, in the east basin, and the Patiti hydrothermal system appear to have been initiated in the aftermath of the eruption, similar to Waimangu Valley to the southwest. Different combinations of turbidity, pH anomalies (both positive and negative), and ORP responses suggest vent fluid compositions vary over short distances within the lake. The seasonal period of stratification limits vertical transport of heat to the surface layer and the hypolimnion temperature of Lake Rotomahana consequently increases with an average warming rate of ~ 0.010 °C/day due to both convective hydrothermal discharge and conductive geothermal heating. A sudden temperature increase occurred during our 2011 survey and was likely the response to an earthquake swarm just 11 days prior.

  19. Neurite outgrowth induced by a synthetic peptide ligand of neural cell adhesion molecule requires fibroblast growth factor receptor activation

    Rønn, L C; Doherty, P; Holm, A;

    2000-01-01

    The neural cell adhesion molecule NCAM is involved in axonal outgrowth and target recognition in the developing nervous system. In vitro, NCAM-NCAM binding has been shown to induce neurite outgrowth, presumably through an activation of fibroblast growth factor receptors (FGFRs). We have recently...... identified a neuritogenic ligand, termed the C3 peptide, of the first immunoglobulin (lg) module of NCAM using a combinatorial library of synthetic peptides. Here we investigate whether stimulation of neurite outgrowth by this synthetic ligand of NCAM involves FGFRs. In primary cultures of cerebellar neurons...... from wild-type mice, the C3 peptide stimulated neurite outgrowth. This response was virtually absent in cultures of cerebellar neurons from transgenic mice expressing a dominant-negative form of the FGFR1. Likewise, in PC12E2 cells transiently expressing a dominant-negative form of the mouse FGFR1...

  20. A dinuclear cobalt complex featuring unprecedented anodic and cathodic redox switches for single-molecule magnet activity.

    Fortier, Skye; Le Roy, Jennifer J; Chen, Chun-Hsing; Vieru, Veacheslav; Murugesu, Muralee; Chibotaru, Liviu F; Mindiola, Daniel J; Caulton, Kenneth G

    2013-10-02

    One-electron oxidation or reduction of the paramagnetic dinuclear Co(II) complex dmp2Nin{Co[N(SiMe3)2]}2 (1; dmp2Nin(2-) = bis(2,6-dimethylphenyl)nindigo), by fully reversible chemical or electrochemical methods, generates the radical salts [1(OEt2)](+) and [1](-), respectively. Full structural and magnetic analyses reveal the locus of the redox changes to be nindigo-based, thus giving rise to ligand-centered radicals sandwiched between two paramagnetic and low-coordinate Co(II) centers. The presence of these sandwiched radicals mediates magnetic coupling between the high-spin (S = 3/2) cobalt ions, which gives rise to single-molecule magnet (SMM) activity in both the oxidized ([1(OEt2)](+)) and reduced ([1](-)) states. This feature represents the first example of a SMM exhibiting fully reversible, dual "ON/OFF" switchability in both the cathodic and anodic states.

  1. A target-based high throughput screen yields Trypanosoma brucei hexokinase small molecule inhibitors with antiparasitic activity.

    Elizabeth R Sharlow

    Full Text Available BACKGROUND: The parasitic protozoan Trypanosoma brucei utilizes glycolysis exclusively for ATP production during infection of the mammalian host. The first step in this metabolic pathway is mediated by hexokinase (TbHK, an enzyme essential to the parasite that transfers the gamma-phospho of ATP to a hexose. Here we describe the identification and confirmation of novel small molecule inhibitors of bacterially expressed TbHK1, one of two TbHKs expressed by T. brucei, using a high throughput screening assay. METHODOLOGY/PRINCIPAL FINDINGS: Exploiting optimized high throughput screening assay procedures, we interrogated 220,233 unique compounds and identified 239 active compounds from which ten small molecules were further characterized. Computation chemical cluster analyses indicated that six compounds were structurally related while the remaining four compounds were classified as unrelated or singletons. All ten compounds were approximately 20-17,000-fold more potent than lonidamine, a previously identified TbHK1 inhibitor. Seven compounds inhibited T. brucei blood stage form parasite growth (0.03activity. None of the compounds displayed structural similarity to known hexokinase inhibitors or human African trypanosomiasis therapeutics. CONCLUSIONS/SIGNIFICANCE: The novel chemotypes identified here could represent leads for future therapeutic development against the African trypanosome.

  2. Molecule nanoweaver

    Gerald, II; Rex E.; Klingler, Robert J.; Rathke, Jerome W.; Diaz, Rocio; Vukovic, Lela

    2009-03-10

    A method, apparatus, and system for constructing uniform macroscopic films with tailored geometric assemblies of molecules on the nanometer scale. The method, apparatus, and system include providing starting molecules of selected character, applying one or more force fields to the molecules to cause them to order and condense with NMR spectra and images being used to monitor progress in creating the desired geometrical assembly and functionality of molecules that comprise the films.

  3. Photogrammetry surveys and mosaic: a useful tool to monitor active zones. Applications to the Indonesian Lusi eruption site.

    Romeo, Giovanni; Di Stefano, Giuseppe; Mazzini, Adriano; Iarocci, Alessandro; Caramelli, Antonio

    2016-04-01

    Unmanned and remotely operated aircraft showed to be an efficient and cost effective way to explore remote or extreme environments. Comparative photogrammetry studies are an efficient way to study and monitor he evolution of geologically active areas and ongoing events and are able to highlight details that are typically lost during traditional field campaigns. The Lusi mud eruption in eastern Java (Indonesia) represents one of the most spectacular geological phenomena that is ongoing since May 2006. In the framework of the Lusi Lab project (ERC grant n° 308126) we designed and constructed a multipurpose drone to survey the eruption site. Among the numerous other payloads, the Lusi drone is equipped with Olympus EPM-2 and Go-Pro Hero3 cameras that allow the operator to collect video stills, high quality pictures and to complete photogrammetry surveys. Targeted areas have been selected for detailed studies in the 7 km2 region inside the embankment that was prevent the mud burial of the settlements in the Sidoarjo Regency. The region is characterized by the presence of the Watukosek fault zone. This strike slip system originates from the Arjuno-Welirang volcanic complex and extends to the north east of the Java Island intersecting the Lusi crater. Therefore of particular interest are the faulted surveyed areas present around the Lusi crater inside the embankment. Results reveal a surprising accuracy for the collected mosaic. Multiple surveys are able to reveal the changes and the evolution of the fault through time and to indicate more active zones. In particular this type of survey can highlight the weakness zones and is thus useful to prevent potential geohazards in the area. The poster shows the aerial survey results, including a 3d-printed slice of LuSi, obtained combining 2500 16 Mp photographs. A 3d zoomed detail is also shown, evidencing the resolution that this technique can offer.

  4. Ground motion parameters of Shillong plateau: One of the most seismically active zones of northeastern India

    Saurabh Baruah; Santanu Baruah; Naba Kumar Gogoi; Olga Erteleva; Felix Aptikaev; J.R.Kayal

    2009-01-01

    Strong ground motion parameters for Shillong plateau of northeastern India are examined. Empirical relations are obtained for main parameters of ground motions as a function of earthquake magnitude, fault type, source depth, velocity characterization of medium and distance. Correlation between ground motion parameters and characteristics of seismogenic zones are established. A new attenuation relation for peak ground acceleration is developed, which predicts higher expected PGA in the region. Parameters of strong motions, particularly the predominant periods and duration of vibrations, depend on the morphology of the studied area. The study measures low estimates of logarithmic width in Shillong plateau. The attenuation relation estimated for pulse width critically indicates increased pulse width dependence on the logarithmic distance which accounts for geometrical spreading and anelastic attenuation.

  5. Mapping bathymetry in an active surf zone with the WorldView2 multispectral satellite

    Trimble, S. M.; Houser, C.; Brander, R.; Chirico, P.

    2015-12-01

    Rip currents are strong, narrow seaward flows of water that originate in the surf zones of many global beaches. They are related to hundreds of international drownings each year, but exact numbers are difficult to calculate due to logistical difficulties in obtaining accurate incident reports. Annual average rip current fatalities are estimated to be ~100, 53 and 21 in the United States (US), Costa Rica, and Australia respectively. Current warning systems (e.g. National Weather Service) do not account for fine resolution nearshore bathymetry because it is difficult to capture. The method shown here could provide frequent, high resolution maps of nearshore bathymetry at a scale required for improved rip prediction and warning. This study demonstrates a method for mapping bathymetry in the surf zone (20m deep and less), specifically within rip channels, because rips form at topographically low spots in the bathymetry as a result of feedback amongst waves, substrate, and antecedent bathymetry. The methods employ the Digital Globe WorldView2 (WV2) multispectral satellite and field measurements of depth to generate maps of the changing bathymetry at two embayed, rip-prone beaches: Playa Cocles, Puerto Viejo de Talamanca, Costa Rica, and Bondi Beach, Sydney, Australia. WV2 has a 1.1 day pass-over rate with 1.84m ground pixel resolution of 8 bands, including 'yellow' (585-625 nm) and 'coastal blue' (400-450 nm). The data is used to classify bottom type and to map depth to the return in multiple bands. The methodology is tested at each site for algorithm consistency between dates, and again for applicability between sites.

  6. The Activity of Major Faults and the Hydrothermal Alteration Zone at Tianchi Volcano of Changbaishan

    Liu Mingjun; Gu Menglin; Sun Zhenguo; Wei Haiquan; Jin Bolu

    2004-01-01

    It is found by field investigation that the near horizontal top surface of the brown or brick-red hydrothermal alteration zone varies obviously in elevation at different sections of the same layer on the caldera's inner wall of Tianchi, with that at the north section near the Tianwen Peak about 110 m higher than that at the south near the Jiangjun Peak in Korea. The top surface of the hydrothermal alteration zone can be taken as key horizon to tectonic movement. The difference indicates that the total uplift height of the NW wall of the Liudaogou-TianchiJingfengshan fault, the principal fault trending NE at Tianchi, is bigger than that of the SE wall ever since the occurrence of hydrothermal alteration. This also explains why the topography in the northwest side of Tianchi is steeper and with more developed river system than in the southeast. The uplifting of the northeastern wall is bigger than that of the southwest along the principal NW-trend fault, namely, the Baishanzhen-Tianchi-Jince fault. It is observed from characters of hydrothermal alteration and the palaeoresiduum, that the recent vertical movement rate along the principal NE-trend fault is larger than that of the principal NW-trend fault. The two faults intersect at Tianchi, dividing the volcano into 4 blocks, with the uplift magnitudes decreasing successively in the order of the north, the west, the east and the south block. The biggest uplift of the north block corresponds well to the shallow magma batch in the north of Tianchi observed by DSS and telluric electromagnetic sounding, and etc.and they may be related with the causes.

  7. Presynaptic calcium channels and α3-integrins are complexed with synaptic cleft laminins, cytoskeletal elements and active zone components.

    Carlson, Steven S; Valdez, Gregorio; Sanes, Joshua R

    2010-11-01

    At chemical synapses, synaptic cleft components interact with elements of the nerve terminal membrane to promote differentiation and regulate function. Laminins containing the β2 subunit are key cleft components, and they act in part by binding the pore-forming subunit of a pre-synaptic voltage-gated calcium channel (Ca(v)α) (Nishimune et al. 2004). In this study, we identify Ca(v)α-associated intracellular proteins that may couple channel-anchoring to assembly or stabilization of neurotransmitter release sites called active zones. Using Ca(v)α-antibodies, we isolated a protein complex from Torpedo electric organ synapses, which resemble neuromuscular junctions but are easier to isolate in bulk. We identified 10 components of the complex: six cytoskeletal proteins (α2/β2 spectrins, plectin 1, AHNAK/desmoyokin, dystrophin, and myosin 1), two active zone components (bassoon and piccolo), synaptic laminin, and a calcium channel β subunit. Immunocytochemistry confirmed these proteins in electric organ synapses, and PCR analysis revealed their expression by developing mammalian motor neurons. Finally, we show that synaptic laminins also interact with pre-synaptic integrins containing the α3 subunit. Together with our previous finding that a distinct synaptic laminin interacts with SV2 on nerve terminals (Son et al. 2000), our results identify three paths by which synaptic cleft laminins can send developmentally important signals to nerve terminals.

  8. Identifying active interplate and intraplate fault zones in the western Caribbean plate from seismic reflection data and the significance of the Pedro Bank fault zone in the tectonic history of the Nicaraguan Rise

    Ott, B.; Mann, P.

    2015-12-01

    The offshore Nicaraguan Rise in the western Caribbean Sea is an approximately 500,000 km2 area of Precambrian to Late Cretaceous tectonic terranes that have been assembled during the Late Cretaceous formation of the Caribbean plate and include: 1) the Chortis block, a continental fragment; 2) the Great Arc of the Caribbean, a deformed Cretaceous arc, and 3) the Caribbean large igneous province formed in late Cretaceous time. Middle Eocene to Recent eastward motion of the Caribbean plate has been largely controlled by strike-slip faulting along the northern Caribbean plate boundary zone that bounds the northern margin of the Nicaraguan Rise. These faults reactivate older rift structures near the island of Jamaica and form the transtensional basins of the Honduran Borderlands near Honduras. Recent GPS studies suggest that small amount of intraplate motion within the current margin of error of GPS measurements (1-3 mm/yr) may occur within the center of the western Caribbean plate at the Pedro Bank fault zone and Hess Escarpment. This study uses a database of over 54,000 km of modern and vintage 2D seismic data, combined with earthquake data and results from previous GPS studies to define the active areas of inter- and intraplate fault zones in the western Caribbean. Intraplate deformation occurs along the 700-km-long Pedro Bank fault zone that traverses the center of the Nicaraguan Rise and reactivates the paleo suture zone between the Great Arc of the Caribbean and the Caribbean large igneous province. The Pedro Bank fault zone also drives active extension at the 200-km-long San Andres rift along the southwest margin of the Nicaraguan Rise. Influence of the Cocos Ridge indentor may be contributing to reactivation of faulting along the southwesternmost, active segment of the Hess Escarpment.

  9. Isolation of a small molecule with anti-MRSA activity from a mangrove symbiont Streptomyces sp. PVRK-1 and its biomedical studies inZebrafish embryos

    Rajaretinam Rajesh Kannan; Appadurai Muthamil Iniyan; Vincent Samuel Gnana Prakash

    2011-01-01

    Objective: The aim of the present study was to isolate the anti-MRSA (Methicillin ResistantStaphylococcus aureus ) molecule from the Mangrove symbiont Streptomyces and its biomedical studies in Zebrafish embryos. Methods: MRSA was isolated from the pus samples of Colachal hospitals and confirmed by amplification of mecA gene. Anti-MRSA molecule producing strain was identified by 16s rRNA gene sequencing. Anti-MRSA compound production was optimized by Solid State Fermentation (SSF) and the purification of the active molecule was carried out by TLC and RP-HPLC. The inhibitory concentration and LC50 were calculated using Statistical software SPSS. The Biomedical studies including the cardiac assay and organ toxicity assessment were carried out in Zebrafish. Results: The bioactive anti-MRSA small molecule A2 was purified by TLC with Rf value of 0.37 with 1.389 retention time at RP-HPLC. The Inhibitory Concentration of the purified molecule A2 was 30 μg/mL but, the inhibitory concentration of the MRSA in the infected embryo was 32-34 μg/mL for TLC purified molecule A2 with LC50 mean value was 61.504 μg/mL. Zebrafish toxicity was assessed in 48-60 μg/mL by observing the physiological deformities and the heart beat rates (HBR) of embryos for anti MRSA molecule showed the mean of 41.33-41.67 HBR/15 seconds for 40 μg/mL and control was 42.33-42.67 for 15 seconds which significantly showed that the anti-MRSA molecule A2 did not affected the HBR. Conclusions:Anti-MRSA molecule from Streptomyces sp PVRK-1 was isolated and biomedical studies in Zebrafish model assessed that the molecule was non toxic at the minimal inhibitory concentration of MRSA.

  10. Direct measurement and characterization of active photosynthesis zones inside biofuel producing and wastewater remediating microalgal biofilms

    Bernstein, Hans C.; Kesaano, Maureen; Moll, Karen; Smith, Terence; Gerlach, Robin; Carlson, Ross; Miller, Charles D.; Peyton, Brent; Cooksey, Keith; Gardner, Robert D.; Sims, Ronald C.

    2014-03-01

    Abstract: Microalgal biofilm based technologies are of keen interest due to their high biomass concentrations and ability to utilize renewable resources, such as light and CO2. While photoautotrophic biofilms have long been used for wastewater remediation applications, biofuel production represents a relatively new and under-represented focus area. However, the direct measurement and characterization of fundamental parameters required for physiological analyses are challenging due to biofilm heterogeneity. This study evaluated oxygenic photosynthesis and biofuel precursor molecule production using a novel rotating algal biofilm reactor (RABR) operated at field- and laboratory-scales for wastewater remediation and biofuel production, respectively. Clear differences in oxygenic-photosynthesis, respiration and biofuel-precursor capacities were observed between the two systems and different conditions based on light and nitrogen availability. Nitrogen depletion was not found to have the same effect on lipid accumulation compared to prior planktonic studies. Physiological characterizations of these microalgal biofilms identify potential areas for future process optimization.

  11. Soluble Collectin-12 (CL-12) Is a Pattern Recognition Molecule Initiating Complement Activation via the Alternative Pathway.

    Ma, Ying Jie; Hein, Estrid; Munthe-Fog, Lea; Skjoedt, Mikkel-Ole; Bayarri-Olmos, Rafael; Romani, Luigina; Garred, Peter

    2015-10-01

    Soluble defense collagens including the collectins play important roles in innate immunity. Recently, a new member of the collectin family named collectin-12 (CL-12 or CL-P1) has been identified. CL-12 is highly expressed in umbilical cord vascular endothelial cells as a transmembrane receptor and may recognize certain bacteria and fungi, leading to opsonophagocytosis. However, based on its structural and functional similarities with soluble collectins, we hypothesized the existence of a fluid-phase analog of CL-12 released from cells, which may function as a soluble pattern-recognition molecule. Using recombinant CL-12 full length or CL-12 extracellular domain, we determined the occurrence of soluble CL-12 shed from in vitro cultured cells. Western blot showed that soluble recombinant CL-12 migrated with a band corresponding to ∼ 120 kDa under reducing conditions, whereas under nonreducing conditions it presented multimeric assembly forms. Immunoprecipitation and Western blot analysis of human umbilical cord plasma enabled identification of a natural soluble form of CL-12 having an electrophoretic mobility pattern close to that of shed soluble recombinant CL-12. Soluble CL-12 could recognize Aspergillus fumigatus partially through the carbohydrate-recognition domain in a Ca(2+)-independent manner. This led to activation of the alternative pathway of complement exclusively via association with properdin on A. fumigatus as validated by detection of C3b deposition and formation of the terminal complement complex. These results demonstrate the existence of CL-12 in a soluble form and indicate a novel mechanism by which the alternative pathway of complement may be triggered directly by a soluble pattern-recognition molecule.

  12. The Restraining Stepovers And Releasing Bends Along The Active East Anatolian Fault Zone, Turkey: Celikhan Region As A Case Study

    Isik, V.; Seyitoglu, G.; Herece, E.; Saber, R.; Caglayan, A.

    2013-12-01

    The Arabia-Eurasia convergence involves intracontinental shortening in SW Turkey. Two active fault zones in Turkey, the North Anatolian Fault Zone (NAFZ) and the East Anatolian Fault Zone (EAFZ), divide the area into the Anatolian micro-plate accommodating SSW-directed movement. The EAFZ is a crustal-scale strike-slip fault, represents a sinistral NE-striking zone, characterized by numerous, complex faults and segmented surface ruptures. The Celikhan region, selected for this study, is located approximately 30 kilometers north of the city of Adiyaman. Although two segments representing the latest seismic activity of the EAFZ have been mapped, named Siro and Erkenek segments, the region contains many segments that become more complex fault pattern. The Celikhan region form part of geology of SE Turkey and is hosted by metamorphic rocks (Poturge metamorphites and Malatya metamorphites), ophiolitic and volcanic units (Kocali complex and Maden complex), limestone with fossils (Midyat formation) and Plio-Quaternary deposits. These rocks are transected by series of steeply dipping to subvertical the EAFZ preserving good indications to sinistral displacement with or/and without normal and reverse component associated with fault rock development. In the northern portion of the study area, the EAFZ is defined by relatively straight, polished recrystallized limestone of the Malatya metamorphites slip surface, which strike N50°-55°E and dip 80°-88° NW. Well-develeoped slickenlines with rakes between 10°-15°, corrugations and some brittle kinematic indicators marks on the slip-surface, indicating strike-slip displacement with minor dip-slip component. This fault create a restraining stepover with the fault near to Celikhan and Akdag can be interpreted as push up structure. At the southwest of Celikhan, the EAFZ create a releasing bend and several strike-slip fault strands with normal component striking N40°-60°E. These subsidiary faults containing centimeter

  13. Photophysical Study of Polymer-Based Solar Cells with an Organo-Boron Molecule in the Active Layer

    Sergio Romero-Servin

    2015-07-01

    Full Text Available Our group previously reported the synthesis of four polythiophene derivatives (P1–P4 used for solar cells. The cells were prepared under room conditions by spin coating, leading to low efficiencies. However, after the addition of 6-nitro-3-(E-3-(4-dimethylaminophenylallylidene-2,3-dihydrobenzo[d]-[1,3,2] oxazaborole (M1 to their active layers, the efficiencies of the cells showed approximately a two-fold improvement. In this paper, we study this enhancement mechanism by performing ultrafast transient absorption (TA experiments on the active layer of the different cells. Our samples consisted of thin films of a mixture of PC61BM with the polythiophenes derivatives P1–P4. We prepared two versions of each sample, one including the molecule M1 and another without it. The TA data suggests that the efficiency improvement after addition of M1 is due not only to an extended absorption spectrum towards the infrared region causing a larger population of excitons but also to the possible creation of additional channels for transport of excitons and/or electrons to the PC61BM interface.

  14. Altered glycosylation of complexed native IgG molecules is associated with disease activity of systemic lupus erythematosus.

    Sjöwall, C; Zapf, J; von Löhneysen, S; Magorivska, I; Biermann, M; Janko, C; Winkler, S; Bilyy, R; Schett, G; Herrmann, M; Muñoz, L E

    2015-05-01

    highest activity of systemic lupus erythematosus. Our results show that native circulating IgG complexes from active systemic lupus erythematosus patients expose fucosyl residues and their glycan core is accessible to soluble lectins. Two putative mechanisms may contribute to the increased exposure of these glycans: (1) the canonical N-glycosylation site of the IgG-CH2 domain; (2) an IgG binding non-IgG molecule, like complement or C-reactive protein. In both cases the complexed IgG may be alternatively targeted to lectin receptors of effector cells, e.g. dendritic cells.

  15. 78 FR 22512 - Foreign-Trade Zone (FTZ) 225-Springfield, Missouri; Notification of Proposed Production Activity...

    2013-04-16

    ... Production Activity; General Dynamics Ordnance and Tactical Systems Munitions Services (Demilitarization of... under Section 400.38 of the Board's regulations. The facility is used for the demilitarization of... could allow GDOTS to conduct demilitarization activity on imported articles at the facility....

  16. Optogenetic stimulation of glutamatergic neuronal activity in the striatum enhances neurogenesis in the subventricular zone of normal and stroke mice.

    Song, Mingke; Yu, Shan Ping; Mohamad, Osama; Cao, Wenyuan; Wei, Zheng Zachory; Gu, Xiaohuan; Jiang, Michael Qize; Wei, Ling

    2017-02-01

    Neurogenesis in the subventricular zone (SVZ) of the adult brain may contribute to tissue repair after brain injuries. Whether SVZ neurogenesis can be upregulated by specific neuronal activity in vivo and promote functional recovery after stroke is largely unknown. Using the spatial and cell type specific optogenetic technique combined with multiple approaches of in vitro, ex vivo and in vivo examinations, we tested the hypothesis that glutamatergic activation in the striatum could upregulate SVZ neurogenesis in the normal and ischemic brain. In transgenic mice expressing the light-gated channelrhodopsin-2 (ChR2) channel in glutamatergic neurons, optogenetic stimulation of the glutamatergic activity in the striatum triggered glutamate release into SVZ region, evoked membrane currents, Ca(2+) influx and increased proliferation of SVZ neuroblasts, mediated by AMPA receptor activation. In ChR2 transgenic mice subjected to focal ischemic stroke, optogenetic stimuli to the striatum started 5days after stroke for 8days not only promoted cell proliferation but also the migration of SVZ neuroblasts into the peri-infarct cortex with increased neuronal differentiation and improved long-term functional recovery. These data provide the first morphological and functional evidence showing a unique striatum-SVZ neuronal regulation via a semi-phasic synaptic mechanism that can boost neurogenic cascades and stroke recovery. The benefits from stimulating endogenous glutamatergic activity suggest a novel regenerative strategy after ischemic stroke and other brain injuries.

  17. Indomethacin treatment reduces microglia activation and increases numbers of neuroblasts in the subventricular zone and ischaemic striatum after focal ischaemia

    ROSANA S LOPES; MARCELO M CARDOSO; ARTHUR O SAMPAIO; MARIO SANTOS BARBOSA Jr; CELICE C SOUZA; MICHELLE C DA SILVA; ELANE MAGNO N FERREIRA; MARCO AURELIOM FREIRE; RAFAEL RODRIGUES LIMA; WALACE GOMES-LEAL

    2016-09-01

    Neuroblasts from the subventricular zone (SVZ) migrate to striatum following stroke, but most of them die inthe ischaemic milieu and this can be related to exacerbated microglial activation. Here, we explored theeffects of the non-steroidal anti-inflammatory indomethacin on microglial activation, neuronal preservation andneuroblast migration following experimental striatal stroke in adult rats. Animals were submitted toendothelin-1 (ET-1)-induced focal striatal ischaemia and were treated with indomethacin or sterile saline(i.p.) for 7 days, being perfused after 8 or 14 days. Immunohistochemistry was performed to assess neuronalloss (anti-NeuN), microglial activation (anti-Iba1, ED1) and migrating neuroblasts (anti-DCX) by countingNeuN, ED1 and DCX-positive cells in the ischaemic striatum or SVZ. Indomethacin treatment reducedmicroglia activation and the number of ED1^{+} cells in both 8 and 14 days post injury as compared withcontrols. There was an increase in the number of DCX^{+} cells in both SVZ and striatum at the same survivaltimes. Moreover, there was a decrease in the number of NeuN^{+} cells in indomethacin-treated animals ascompared with the control group at 8 days but not after 14 days post injury. Our results suggest thatindomethacin treatment modulates microglia activation, contributing to increased neuroblast proliferation inthe SVZ and migration to the ischaemic striatum following stroke.

  18. Breast cancer cell migration is regulated through junctional adhesion molecule-A-mediated activation of Rap1 GTPase.

    McSherry, Elaine A

    2011-03-23

    ABSTRACT: INTRODUCTION: The adhesion protein junctional adhesion molecule-A (JAM-A) regulates epithelial cell morphology and migration, and its over-expression has recently been linked with increased risk of metastasis in breast cancer patients. As cell migration is an early requirement for tumor metastasis, we sought to identify the JAM-A signalling events regulating migration in breast cancer cells. METHODS: MCF7 breast cancer cells (which express high endogenous levels of JAM-A) and primary cultures from breast cancer patients were used for this study. JAM-A was knocked down in MCF7 cells using siRNA to determine the consequences for cell adhesion, cell migration and the protein expression of various integrin subunits. As we had previously demonstrated a link between the expression of JAM-A and β1-integrin, we examined activation of the β1-integrin regulator Rap1 GTPase in response to JAM-A knockdown or functional antagonism. To test whether JAM-A, Rap1 and β1-integrin lie in a linear pathway, we tested functional inhibitors of all three proteins separately or together in migration assays. Finally we performed immunoprecipitations in MCF7 cells and primary breast cells to determine the binding partners connecting JAM-A to Rap1 activation. RESULTS: JAM-A knockdown in MCF7 breast cancer cells reduced adhesion to, and migration through, the β1-integrin substrate fibronectin. This was accompanied by reduced protein expression of β1-integrin and its binding partners αV- and α5-integrin. Rap1 activity was reduced in response to JAM-A knockdown or inhibition, and pharmacological inhibition of Rap1 reduced MCF7 cell migration. No additive anti-migratory effect was observed in response to simultaneous inhibition of JAM-A, Rap1 and β1-integrin, suggesting that they lie in a linear migratory pathway. Finally, in an attempt to elucidate the binding partners putatively linking JAM-A to Rap1 activation, we have demonstrated the formation of a complex between JAM-A, AF

  19. Breast cancer cell migration is regulated through junctional adhesion molecule-A-mediated activation of Rap1 GTPase

    McSherry, Elaine A

    2011-03-23

    Abstract Introduction The adhesion protein junctional adhesion molecule-A (JAM-A) regulates epithelial cell morphology and migration, and its over-expression has recently been linked with increased risk of metastasis in breast cancer patients. As cell migration is an early requirement for tumor metastasis, we sought to identify the JAM-A signalling events regulating migration in breast cancer cells. Methods MCF7 breast cancer cells (which express high endogenous levels of JAM-A) and primary cultures from breast cancer patients were used for this study. JAM-A was knocked down in MCF7 cells using siRNA to determine the consequences for cell adhesion, cell migration and the protein expression of various integrin subunits. As we had previously demonstrated a link between the expression of JAM-A and β1-integrin, we examined activation of the β1-integrin regulator Rap1 GTPase in response to JAM-A knockdown or functional antagonism. To test whether JAM-A, Rap1 and β1-integrin lie in a linear pathway, we tested functional inhibitors of all three proteins separately or together in migration assays. Finally we performed immunoprecipitations in MCF7 cells and primary breast cells to determine the binding partners connecting JAM-A to Rap1 activation. Results JAM-A knockdown in MCF7 breast cancer cells reduced adhesion to, and migration through, the β1-integrin substrate fibronectin. This was accompanied by reduced protein expression of β1-integrin and its binding partners αV- and α5-integrin. Rap1 activity was reduced in response to JAM-A knockdown or inhibition, and pharmacological inhibition of Rap1 reduced MCF7 cell migration. No additive anti-migratory effect was observed in response to simultaneous inhibition of JAM-A, Rap1 and β1-integrin, suggesting that they lie in a linear migratory pathway. Finally, in an attempt to elucidate the binding partners putatively linking JAM-A to Rap1 activation, we have demonstrated the formation of a complex between JAM-A, AF-6

  20. Breast cancer cell migration is regulated through junctional adhesion molecule-A-mediated activation of Rap1 GTPase.

    McSherry, Elaine A

    2012-02-01

    INTRODUCTION: The adhesion protein junctional adhesion molecule-A (JAM-A) regulates epithelial cell morphology and migration, and its over-expression has recently been linked with increased risk of metastasis in breast cancer patients. As cell migration is an early requirement for tumor metastasis, we sought to identify the JAM-A signalling events regulating migration in breast cancer cells. METHODS: MCF7 breast cancer cells (which express high endogenous levels of JAM-A) and primary cultures from breast cancer patients were used for this study. JAM-A was knocked down in MCF7 cells using siRNA to determine the consequences for cell adhesion, cell migration and the protein expression of various integrin subunits. As we had previously demonstrated a link between the expression of JAM-A and beta1-integrin, we examined activation of the beta1-integrin regulator Rap1 GTPase in response to JAM-A knockdown or functional antagonism. To test whether JAM-A, Rap1 and beta1-integrin lie in a linear pathway, we tested functional inhibitors of all three proteins separately or together in migration assays. Finally we performed immunoprecipitations in MCF7 cells and primary breast cells to determine the binding partners connecting JAM-A to Rap1 activation. RESULTS: JAM-A knockdown in MCF7 breast cancer cells reduced adhesion to, and migration through, the beta1-integrin substrate fibronectin. This was accompanied by reduced protein expression of beta1-integrin and its binding partners alphaV- and alpha5-integrin. Rap1 activity was reduced in response to JAM-A knockdown or inhibition, and pharmacological inhibition of Rap1 reduced MCF7 cell migration. No additive anti-migratory effect was observed in response to simultaneous inhibition of JAM-A, Rap1 and beta1-integrin, suggesting that they lie in a linear migratory pathway. Finally, in an attempt to elucidate the binding partners putatively linking JAM-A to Rap1 activation, we have demonstrated the formation of a complex between

  1. Activation of Relaxin Family Receptor 1 from different mammalian species by relaxin peptide and small molecule agonist ML290

    Zaohua eHuang

    2015-08-01

    Full Text Available Relaxin peptide (RLN, which signals through the relaxin family peptide 1 (RXFP1 GPCR receptor, has shown therapeutic effects in an acute heart failure clinical trial. We have identified a small molecule agonist of human RXFP1, ML290; however, it does not activate the mouse receptor. To find a suitable animal model for ML290 testing and to gain mechanistic insights into the interaction of various ligands with RXFP1, we have cloned rhesus macaque, pig, rabbit, and guinea pig RXFP1s and analyzed their activation by RLN and ML290. HEK293T cells expressing macaque or pig RXFP1 responded to relaxin and ML290 treatment as measured by an increase of cAMP production. Guinea pig RXFP1 responded to relaxin but had very low response to ML290 treatment only at highest concentrations used. The rabbit RXFP1 amino acid sequence was the most divergent, with a number of unique substitutions within the ectodomain and the 7-transmembrane domain (7TM. Two splice variants of rabbit RXFP1 derived through alternative splicing of the forth exon were identified. In contrast to the other species, rabbit RXFP1s were activated by ML290, but not with human, pig, mouse, or rabbit relaxins. Using FLAG-tagged constructs, we have shown that both rabbit RXFP1 variants are expressed on the cell surface. No binding of human Eu-labeled relaxin to rabbit RXFP1 was detected, suggesting that in this species RXFP1 might be non-functional. We used chimeric rabbit-human and guinea pig-human constructs to identify regions important for RLN or ML290 receptor activation. Chimeras with the human ectodomain and rabbit 7TM domain were activated by RLN, whereas substitution of part of the guinea pig 7TM domain with the human sequence only partially restored ML290 activation, confirming the allosteric mode of action for the two ligands. Our data demonstrate that macaque and pig models can be used for ML290 testing.

  2. Active crustal deformation of the El Salvador Fault Zone (ESFZ) using GPS data: Implications in seismic hazard assessment

    Staller, Alejandra; Benito, Belen; Jesús Martínez-Díaz, José; Hernández, Douglas; Hernández-Rey, Román; Alonso-Henar, Jorge

    2014-05-01

    El Salvador, Central America, is part of the Chortis block in the northwestern boundary of the Caribbean plate. This block is interacting with a diffuse triple junction point with the Cocos and North American plates. Among the structures that cut the Miocene to Pleistocene volcanic deposits stands out the El Salvador Fault Zone (ESFZ): It is oriented in N90º-100ºE direction, and it is composed of several structural segments that deform Quaternary deposits with right-lateral and oblique slip motions. The ESFZ is seismically active and capable of producing earthquakes such as the February 13, 2001 with Mw 6.6 (Martínez-Díaz et al., 2004), that seriously affected the population, leaving many casualties. This structure plays an important role in the tectonics of the Chortis block, since its motion is directly related to the drift of the Caribbean plate to the east and not with the partitioning of the deformation of the Cocos subduction (here not coupled) (Álvarez-Gómez et al., 2008). Together with the volcanic arc of El Salvador, this zone constitutes a weakness area that allows the motion of forearc block toward the NW. The geometry and the degree of activity of the ESFZ are not studied enough. However their knowledge is essential to understand the seismic hazard associated to this important seismogenic structure. For this reason, since 2007 a GPS dense network was established along the ESFZ (ZFESNet) in order to obtain GPS velocity measurements which are later used to explain the nature of strain accumulation on major faults along the ESFZ. The current work aims at understanding active crustal deformation of the ESFZ through kinematic model. The results provide significant information to be included in a new estimation of seismic hazard taking into account the major structures in ESFZ.

  3. Pseudomonas aeruginosa Biofilm Response and Resistance to Cold Atmospheric Pressure Plasma Is Linked to the Redox-Active Molecule Phenazine.

    Mai-Prochnow, Anne; Bradbury, Mark; Ostrikov, Kostya; Murphy, Anthony B

    2015-01-01

    Pseudomonas aeruginosa is an important opportunistic pathogen displaying high antibiotic resistance. Its resistance is in part due to its outstanding ability to form biofilms on a range of biotic and abiotic surfaces leading to difficult-to-treat, often long-term infections. Cold atmospheric plasma (CAP) is a new, promising antibacterial treatment to combat antibiotic-resistant bacteria. Plasma is ionized gas that has antibacterial properties through the generation of a mix of reactive oxygen and nitrogen species (RONS), excited molecules, charged particles and UV photons. Our results show the efficient removal of P. aeruginosa biofilms using a plasma jet (kINPen med), with no viable cells detected after 5 min treatment and no attached biofilm cells visible with confocal microscopy after 10 min plasma treatment. Because of its multi-factorial action, it is widely presumed that the development of bacterial resistance to plasma is unlikely. However, our results indicate that a short plasma treatment (3 min) may lead to the emergence of a small number of surviving cells exhibiting enhanced resistance to subsequent plasma exposure. Interestingly, these cells also exhibited a higher degree of resistance to hydrogen peroxide. Whole genome comparison between surviving cells and control cells revealed 10 distinct polymorphic regions, including four belonging to the redox active, antibiotic pigment phenazine. Subsequently, the interaction between phenazine production and CAP resistance was demonstrated in biofilms of transposon mutants disrupted in different phenazine pathway genes which exhibited significantly altered sensitivity to CAP.

  4. Resonant activation in polymer translocation: new insights into the escape dynamics of molecules driven by an oscillating field.

    Pizzolato, N; Fiasconaro, A; Adorno, D Persano; Spagnolo, B

    2010-08-04

    The translocation of molecules across cellular membranes or through synthetic nanopores is strongly affected by thermal fluctuations. In this work we study how the dynamics of a polymer in a noisy environment changes when the translocation process is driven by an oscillating electric field. An improved version of the Rouse model for a flexible polymer has been adopted to mimic the molecular dynamics, by taking into account the harmonic interactions between adjacent monomers and the excluded-volume effect by introducing a Lennard-Jones potential between all beads. A bending recoil torque has also been included in our model. The polymer dynamics is simulated in a two-dimensional domain by numerically solving the Langevin equations of motion. Thermal fluctuations are taken into account by introducing a Gaussian uncorrelated noise. The mean first translocation time of the polymer centre of inertia shows a minimum as a function of the frequency of the oscillating forcing field. This finding represents the first evidence of the resonant activation behaviour in the dynamics of polymer translocation.

  5. Polyether ionophores: broad-spectrum and promising biologically active molecules for the control of drug-resistant bacteria and parasites

    Kevin, Dion A; Meujo, Damaris AF; Hamann, Mark T

    2016-01-01

    Background As multidrug-resistant (MDR) pathogens continue to emerge, there is a substantial amount of pressure to identify new drug candidates. Carboxyl polyethers, also referred to as polyether antibiotics, are a unique class of compounds with outstanding potency against a variety of critical infectious disease targets including protozoa, bacteria and viruses. The characteristics of these molecules that are of key interest are their selectivity and high potency against several MDR etiological agents. Objective Although many studies have been published about carboxyl polyether antibiotics, there are no recent reviews of this class of drugs. The purpose of this review is to provide the reader with an overview of the spectrum of activity of polyether antibiotics, their mechanism of action, toxicity and potential as drug candidates to combat drug-resistant infectious diseases. Conclusion Polyether ionophores show a high degree of promise for the potential control of drug-resistant bacterial and parasitic infections. Despite the long history of use of this class of drugs, very limited medicinal chemistry and drug optimization studies have been reported, thus leaving the door open to these opportunities in the future. Scifinder and PubMed were the main search engines used to locate articles relevant to the topic presented in the present review. Keywords used in our search were specific names of each of the 88 compounds presented in the review as well as more general terms such as polyethers, ionophores, carboxylic polyethers and polyether antibiotics. PMID:23480512

  6. 活化白细胞黏附分子与肿瘤%Activated leukocyte cell adhesion molecule and minors

    柯少迎; 庄建良; 潘群雄

    2009-01-01

    Activated leukocyte cell adhesion molecule (ALCAM) is a member of immunoglobulin gene superfamily. It is manily expressed in leucocytes and thymic epithelial cells. ALCAM mediates both heterophilic (ALCAM-CD6) and homophilic(ALCAM-ALCAM) cell-cell interactions. Studies show that ALCAM is highly expressed on multiple tumor cells and plays a role in tumorigenesis and tumor progression. ALCAM expression is closely correlated with several human tumors, including breast cancer, esophageal carcinoma, colon carcinoma, o-varian caneer,hepatocellular carcinoma and malignant melanoma.%活化自细胞黏附分子(ALCAM)是免疫球蛋白基因超家族成员,主要表达在白细胞和胸腺上皮细胞.ALCAM在细胞间异嗜性黏附和同嗜性黏附中起重要作用.研究表明,ALCAM在多种肿瘤细胞表面高表达,参与肿瘤的发生发展,在人类肿瘤,ALCAM的表达与乳腺癌、食管癌、结肠癌、卵巢癌、肝癌及恶性黑色素瘤等密切相关.

  7. Pseudomonas aeruginosa Biofilm Response and Resistance to Cold Atmospheric Pressure Plasma Is Linked to the Redox-Active Molecule Phenazine.

    Anne Mai-Prochnow

    Full Text Available Pseudomonas aeruginosa is an important opportunistic pathogen displaying high antibiotic resistance. Its resistance is in part due to its outstanding ability to form biofilms on a range of biotic and abiotic surfaces leading to difficult-to-treat, often long-term infections. Cold atmospheric plasma (CAP is a new, promising antibacterial treatment to combat antibiotic-resistant bacteria. Plasma is ionized gas that has antibacterial properties through the generation of a mix of reactive oxygen and nitrogen species (RONS, excited molecules, charged particles and UV photons. Our results show the efficient removal of P. aeruginosa biofilms using a plasma jet (kINPen med, with no viable cells detected after 5 min treatment and no attached biofilm cells visible with confocal microscopy after 10 min plasma treatment. Because of its multi-factorial action, it is widely presumed that the development of bacterial resistance to plasma is unlikely. However, our results indicate that a short plasma treatment (3 min may lead to the emergence of a small number of surviving cells exhibiting enhanced resistance to subsequent plasma exposure. Interestingly, these cells also exhibited a higher degree of resistance to hydrogen peroxide. Whole genome comparison between surviving cells and control cells revealed 10 distinct polymorphic regions, including four belonging to the redox active, antibiotic pigment phenazine. Subsequently, the interaction between phenazine production and CAP resistance was demonstrated in biofilms of transposon mutants disrupted in different phenazine pathway genes which exhibited significantly altered sensitivity to CAP.

  8. Strontium-90 activity concentration in soil samples from the exclusion zone of the Fukushima daiichi nuclear power plant

    Sahoo, Sarata Kumar; Kavasi, Norbert; Sorimachi, Atsuyuki; Arae, Hideki; Tokonami, Shinji; Mietelski, Jerzy Wojciech; Łokas, Edyta; Yoshida, Satoshi

    2016-04-01

    The radioactive fission product 90Sr has a long biological half-life (˜18 y) in the human body. Due to its chemical similarity to calcium it accumulates in bones and irradiates the bone marrow, causing its high radio-toxicity. Assessing 90Sr is therefore extremely important in case of a nuclear disaster. In this work 16 soil samples were collected from the exclusion zone (Fukushima Daiichi nuclear power plant, to measure 90Sr activity concentration using liquid scintillation counting. 137Cs activity concentration was also measured with gamma-spectroscopy in order to investigate correlation with 90Sr. The 90Sr activity concentrations ranged from 3.0 ± 0.3 to 23.3 ± 1.5 Bq kg-1 while the 137Cs from 0.7 ± 0.1 to 110.8 ± 0.3 kBq kg-1. The fact that radioactive contamination originated from the Fukushima nuclear accident was obvious due to the presence of 134Cs. However, 90Sr contamination was not confirmed in all samples although detectable amounts of 90Sr can be expected in Japanese soils, as a background, stemming from global fallout due to the atmospheric nuclear weapon tests. Correlation analysis between 90Sr and 137Cs activity concentrations provides a potentially powerful tool to discriminate background 90Sr level from its Fukushima contribution.

  9. Exploring the Cross-sectional Association between Transit-Oriented Development Zoning and Active Travel and Transit Usage in the United States, 2010-2014

    Emily eThrun

    2016-06-01

    Full Text Available Background: In response to traditional zoning codes that contribute to car-dependent, sprawling, and disconnected neighborhoods, communities are reforming their land use laws to create pedestrian-friendly areas that promote physical activity. One such reform is the adoption of transit-oriented developments or districts (TODs. TODs are higher-density, compact, mixed use areas located around transit stops that are designed to encourage walking.Purpose: To identify the characteristics of communities that have adopted TODs in their land use laws and examine if communities that have included TODs in their zoning codes are more likely to have adults that commute by any form of active transportation (i.e., walking, biking, or public transportation or by using public transportation specifically.Methods: Zoning codes effective as of 2010 were obtained for a purposeful sample of the largest 3,914 municipal jurisdictions located in 473 of the most populous US counties and consolidated cities within 48 states and the District of Columbia. They were evaluated to determine whether they included TOD districts or regulations using a coding tool developed by the study team. Descriptive statistics together with t-tests and Pearson’s chi-squared independence test were used to compare characteristics of jurisdictions with and without TOD zoning. Multivariate linear regressions were used to compute the adjusted association between TOD zoning and taking public or active transportation to work.Results: Jurisdictions with TOD zoning were located more in the South and West than non-TOD jurisdictions and were more populous, higher income, more racially diverse, and younger. Jurisdictions with TOD zoning had significantly higher percentages of occupied housing with no vehicle than those without TOD zoning. TOD zoning was associated with significantly higher rates of public transportation to work (β=2.10, 95% CI=0.88, 3.32 and active transportation to work (β=2.48, 95

  10. Characterization Activities to Determine the Extent of DNAPL in the Vadose Zone at the A-014 Outfall of A/M Area

    Jackson, D.G.

    2000-09-05

    The purpose of this investigation was to perform characterization activities necessary to confirm the presence and extent of DNAPL in the shallow vadose zone near the headwaters of the A-014 Outfall. Following the characterization, additional soil vapor extraction wells and vadose monitoring probes were installed to promote and monitor remediation activities in regions of identified DNAPL.

  11. Activated endothelial interleukin-1beta, -6, and -8 concentrations and intercellular adhesion molecule-1 expression are attenuated by lidocaine.

    Lan, Wei

    2012-02-03

    Endothelial cells play a key role in ischemia reperfusion injury. We investigated the effects of lidocaine on activated human umbilical vein endothelial cell (HUVEC) interleukin (IL)-1beta, IL-6, and IL-8 concentrations and intercellular adhesion molecule-1 (ICAM-1) expression. HUVECs were pretreated with different concentrations of lidocaine (0 to 0.5 mg\\/mL) for 60 min, thereafter tumor necrosis factor-alpha was added at a concentration of 2.5 ng\\/mL and the cells incubated for 4 h. Supernatants were harvested, and cytokine concentrations were analyzed by enzyme-linked immunosorbent assay. Endothelial ICAM-1 expression was analyzed by using flow cytometry. Differences were assessed using analysis of variance and post hoc unpaired Student\\'s t-test where appropriate. Lidocaine (0.5 mg\\/mL) decreased IL-1beta (1.89 +\\/- 0.11 versus 4.16 +\\/- 1.27 pg\\/mL; P = 0.009), IL-6 (65.5 +\\/- 5.14 versus 162 +\\/- 11.5 pg\\/mL; P < 0.001), and IL-8 (3869 +\\/- 785 versus 14,961 +\\/- 406 pg\\/mL; P < 0.001) concentrations compared with the control. IL-1beta, IL-6, and IL-8 concentrations in HUVECs treated with clinically relevant plasma concentrations of lidocaine (0.005 mg\\/mL) were similar to control. ICAM-1 expression on lidocaine-treated (0.05 mg\\/mL) HUVECs was less than on controls (198 +\\/- 52.7 versus 298 +\\/- 50.3; Mean Channel Fluorescence; P < 0.001). Activated endothelial IL-1beta, IL-6, and IL-8 concentrations and ICAM-1 expression are attenuated only by lidocaine at concentrations larger than clinically relevant concentrations.

  12. 78 FR 49254 - Foreign-Trade Zone 158-Vicksburg/Jackson, Mississippi; Authorization of Production Activity...

    2013-08-13

    ... Production Activity; Extension of Production Authority; Bauhaus USA, Inc. (Upholstered Furniture); Saltillo... Bauhaus USA, Inc., in Saltillo, Mississippi. The notification was processed in accordance with the... foreign status (19 CFR 146.42) is limited to 3.5 million square yards. 2. Bauhaus USA, Inc., must...

  13. 78 FR 24158 - Foreign-Trade Zone (FTZ) 75-Phoenix, Arizona; Notification of Proposed Production Activity...

    2013-04-24

    ... Production Activity; Orbital Sciences Corporation (Satellites and Spacecraft Launch Vehicles); Gilbert... on behalf of Orbital Sciences Corporation (OSC), located in Gilbert, Arizona. The notification... (202) 482-1378. Dated: April 17, 2013. Andrew McGilvray, Executive Secretary. BILLING CODE P...

  14. 78 FR 72861 - Foreign-Trade Zone (FTZ) 20-Suffolk, Virginia, Notification of Proposed Production Activity...

    2013-12-04

    ... portable light towers and diesel-powered generator sets for residential, commercial, and industrial applications. Pursuant to 15 CFR 400.14(b), FTZ activity would be limited to the specific foreign-status... assemblies; guide pulleys; winches; axles; tire-rim assemblies; clips; pins; brackets; bolts; junction...

  15. Discovery and Characterization of a Cell-Permeable, Small-Molecule c-Abl Kinase Activator that Binds to the Myristoyl Binding Site

    Yang, Jingsong; Campobasso, Nino; Biju, Mangatt P.; Fisher, Kelly; Pan, Xiao-Qing; Cottom, Josh; Galbraith, Sarah; Ho, Thau; Zhang, Hong; Hong, Xuan; Ward, Paris; Hofmann, Glenn; Siegfried, Brett; Zappacosta, Francesca; Washio, Yoshiaki; Cao, Ping; Qu, Junya; Bertrand, Sophie; Wang, Da-Yuan; Head, Martha S.; Li, Hu; Moores, Sheri; Lai, Zhihong; Johanson, Kyung; Burton, George; Erickson-Miller, Connie; Simpson, Graham; Tummino, Peter; Copeland, Robert A.; Oliff, Allen (GSKPA)

    2014-10-02

    c-Abl kinase activity is regulated by a unique mechanism involving the formation of an autoinhibited conformation in which the N-terminal myristoyl group binds intramolecularly to the myristoyl binding site on the kinase domain and induces the bending of the {alpha}I helix that creates a docking surface for the SH2 domain. Here, we report a small-molecule c-Abl activator, DPH, that displays potent enzymatic and cellular activity in stimulating c-Abl activation. Structural analyses indicate that DPH binds to the myristoyl binding site and prevents the formation of the bent conformation of the {alpha}I helix through steric hindrance, a mode of action distinct from the previously identified allosteric c-Abl inhibitor, GNF-2, that also binds to the myristoyl binding site. DPH represents the first cell-permeable, small-molecule tool compound for c-Abl activation.

  16. Electron correlation effects in enhanced-ionization of molecules:A time-dependent generalized-active-space configuration-interaction study

    Chattopadhyay, S; Madsen, L B

    2015-01-01

    We numerically study models of $\\mathrm{H}_2$ and $\\mathrm{LiH}$ molecules, aligned collinearly with the linear polarization of the external field, to elucidate the possible role of correlation in the enhanced-ionization phenomena. Correlation is considered at different levels of approximation with the time-dependent generalized-active-space configuration-interaction method. The results of our studies show that enhanced ionization occurs in multielectron molecules, that correlation is important and they also demonstrate significant deviations between the results of the single-active-electron approximation and more accurate configuration-interaction methods. With the inclusion of correlation we show strong carrier-envelope-phase effects in the enhanced ionization of the asymmetric heteronuclear $\\mathrm{LiH}$-like molecule. The correlated calculation shows an intriguing feature of cross-over in enhanced ionization with two carrier-envelope-phases at critical inter-nuclear separation.

  17. Examining the antimicrobial activity and toxicity to animal cells of different types of CO-releasing molecules.

    Nobre, Lígia S; Jeremias, Hélia; Romão, Carlos C; Saraiva, Lígia M

    2016-01-28

    Transition metal carbonyl complexes used as CO-releasing molecules (CORMs) for biological and therapeutic applications may exhibit interesting antimicrobial activity. However, understanding the chemical traits and mechanisms of action that rule this activity is required to establish a rationale for the development of CORMs into useful antibiotics. In this work the bactericidal activity, the toxicity to eukaryotic cells, and the ability of CORMs to deliver CO to bacterial and eukaryotic cells were analysed for a set of seven CORMs that differ in the transition metal, ancillary ligands and the CO release profile. Most of these CORMs exhibited bactericidal properties that decrease in the following order: CORM-2 > CORM-3 > ALF062 > ALF850 > ALF186 > ALF153 > [Fe(SBPy3)(CO)](BF4)2. A similar yet not entirely coincident decreasing order was found for their induction of intracellular reactive oxygen species (ROS) in E. coli. In contrast, studies in model animal cells showed that for any given CORM, the level of intracellular ROS generated was negligible when compared with that measured inside bacteria. Importantly, these CORMs were in general not toxic to eukaryotic cells, namely murine macrophages, kidney LLC-PK1 epithelial cells, and liver cell line HepG2. CORM-2 and CORM-3 delivered CO to the intracellular space of both E. coli and the two types of tested eukaryotic cells, yet toxicity was only elicited in the case of E. coli. CO delivered by ALF186 into the intercellular space did not enter E. coli cells and the compound was not toxic to either bacteria or to eukaryotic cells. The Fe(ii) carbonyl complex [Fe(SBPy3)(CO)](2+) had the reverse, undesirable toxicity profile, being unexpectedly toxic to eukaryotic cells and non-toxic to E. coli. ALF153, the most stable complex in the whole set, was essentially devoid of toxicity or ROS induction ability in all cells. These results suggest that CORMs have a relevant therapeutic potential as antimicrobial drugs since (i) they

  18. A NEW PERSPECTIVE OF THE RADIO BRIGHT ZONE AT THE GALACTIC CENTER: FEEDBACK FROM NUCLEAR ACTIVITIES

    Zhao, Jun-Hui [Harvard-Smithsonian Center for Astrophysics, 60 Garden Street, Cambridge, MA 02138 (United States); Morris, Mark R. [Department of Physics and Astronomy, University of California Los Angeles, Los Angeles, CA 90095 (United States); Goss, W. M., E-mail: jzhao@cfa.harvard.edu [NRAO, P.O. Box O, Socorro, NM 87801 (United States)

    2016-02-01

    New observations of Sgr A have been carried out with the Jansky VLA in the B and C arrays using the broadband (2 GHz) continuum mode at 5.5 GHz. The field of view covers the central 13′ (30 pc) region of the radio-bright zone at the Galactic center. Using the multi-scale and multi-frequency-synthesis (MS-MFS) algorithms in CASA, we have imaged Sgr A with a resolution of 1″, achieving an rms noise of 8 μJy beam{sup −1}, and a dynamic range of 100,000:1. Both previously known and newly identified radio features in this region are revealed, including numerous filamentary sources. The radio continuum image is compared with Chandra X-ray images, with a CN emission-line image obtained with the Submillimeter Array and with detailed Paschen-α images obtained with Hubble Space Telescope/NICMOS. We discuss several prominent features in the radio image. The “Sgr A west Wings” extend 2′ (5 pc) from the NW and SE tips of the Sgr A west H ii region (the “Mini-spiral”) to positions located 2.9 and 2.4 arcmin to the northwest and southeast of Sgr A*, respectively. The NW wing, along with several other prominent features, including the previously identified “NW Streamers,” form an elongated radio lobe (NW lobe), oriented nearly perpendicular to the Galactic plane. This radio lobe, with a size of 6.′3 × 3.′2 (14.4 pc × 7.3 pc), has a known X-ray counterpart. In the outer region of the NW lobe, a row of three thermally emitting rings is observed. A field containing numerous amorphous radio blobs extends for a distance of ∼2 arcmin beyond the tip of the SE wing; these newly recognized features coincide with the SE X-ray lobe. Most of the amorphous radio blobs in the NW and SE lobes have Paschen-α counterparts. We propose that they have been produced by shock interaction of ambient gas concentrations with a collimated nuclear wind or an outflow that originated from within the circumnuclear disk (CND). We also discuss the possibility that the ionized

  19. A New Perspective of the Radio Bright Zone at The Galactic Center: Feedback from Nuclear Activities

    Zhao, Jun-Hui; Morris, Mark R.; Goss, W. M.

    2016-02-01

    New observations of Sgr A have been carried out with the Jansky VLA in the B and C arrays using the broadband (2 GHz) continuum mode at 5.5 GHz. The field of view covers the central 13‧ (30 pc) region of the radio-bright zone at the Galactic center. Using the multi-scale and multi-frequency-synthesis (MS-MFS) algorithms in CASA, we have imaged Sgr A with a resolution of 1″, achieving an rms noise of 8 μJy beam-1, and a dynamic range of 100,000:1. Both previously known and newly identified radio features in this region are revealed, including numerous filamentary sources. The radio continuum image is compared with Chandra X-ray images, with a CN emission-line image obtained with the Submillimeter Array and with detailed Paschen-α images obtained with Hubble Space Telescope/NICMOS. We discuss several prominent features in the radio image. The “Sgr A west Wings” extend 2‧ (5 pc) from the NW and SE tips of the Sgr A west H ii region (the “Mini-spiral”) to positions located 2.9 and 2.4 arcmin to the northwest and southeast of Sgr A*, respectively. The NW wing, along with several other prominent features, including the previously identified “NW Streamers,” form an elongated radio lobe (NW lobe), oriented nearly perpendicular to the Galactic plane. This radio lobe, with a size of 6.‧3 × 3.‧2 (14.4 pc × 7.3 pc), has a known X-ray counterpart. In the outer region of the NW lobe, a row of three thermally emitting rings is observed. A field containing numerous amorphous radio blobs extends for a distance of ˜2 arcmin beyond the tip of the SE wing; these newly recognized features coincide with the SE X-ray lobe. Most of the amorphous radio blobs in the NW and SE lobes have Paschen-α counterparts. We propose that they have been produced by shock interaction of ambient gas concentrations with a collimated nuclear wind or an outflow that originated from within the circumnuclear disk (CND). We also discuss the possibility that the ionized wind or

  20. Military Activities in the Exclusive Economic Zone: Preventing Uncertainty and Defusing Conflict

    2002-03-01

    freedom of navigation." Id. (citing Edmund V. Carreno, America Latina y los Problems Contemporaneos del Derecho del Mar 27-28 (Santiago, 1973)). But see... fiscal , customs, immigration, and health laws.薯 As discussed above, however, despite this diplomatic and operational activity, while some States have...the coastal State may exercise the control necessary to.. prevent infringement of its customs, fiscal , inmnigration or sanitary laws and regulations

  1. Preliminary Assessment of the Nuclide Migration from the Activation Zone Around the Proposed Spallation Neutron Source Facility

    Dole, L.R.

    1998-09-01

    The purpose of this study is to investigate the potential impacts of migrating radionuclides from the activation zone around the proposed Spallation Neutron Source (SNS). Using conservatively high estimates of the potential inventory of radioactive activation products that could form in the proposed compacted-soil shield berm around an SNS facility on the Oak Ridge Reservation (ORR), a conservative, simplified transport model was used to estimate the potential worst-case concentrations of the 12 long-lived isotopes in the groundwater under a site with the hydrologic characteristics of the ORR. Of the 12, only 3 isotopes showed any potential to exceed the U.S. Nuclear Regulatory Commission (NRC) 10 Code of Federal Regulations (CFR) Part 20 Drinking Water Limits (DWLs). These isotopes were 14C, 22Na, and 54Mn. The latter two activation products have very short half-lives of 2.6 years and 0.854 year, respectively. Therefore, these will decay before reaching an off-site receptor, and they cannot pose off-site hazards. However, for this extremely conservative model, which overestimates the mobility of the contaminant, 14C, which has a 5,730-year half-life, was shown to represent a potential concern in the context of this study's conservative assumptions. This study examines alternative modifications to the SNS shield berm and makes recommendations.

  2. Multilayer stress from gravity and its tectonic implications in urban active fault zone: A case study in Shenzhen, South China

    Xu, Chuang; Wang, Hai-hong; Luo, Zhi-cai; Ning, Jin-sheng; Liu, Hua-liang

    2015-03-01

    It is significant to identify urban active faults for human life and social sustainable development. The ordinary methods to detect active faults, such as geological survey, artificial seismic exploration, and electromagnetic exploration, are not convenient to be carried out in urban area with dense buildings. It is also difficult to supply information about vertical extension of the deeper faults by these methods. Gravity, reflecting the mass distribution of the Earth's interior, provides an alternative way to detect faults, which is more efficient and convenient for urban active fault detection than the aforementioned techniques. Based on the multi-scale decomposition of gravity anomalies, a novel method to invert multilayer horizontal tectonic stresses is proposed. The inverted multilayer stress fields are further used to infer the distribution and stability of the main faults. In order to validate our method, the multilayer stress fields in the Shenzhen fault zone are calculated as a case study. The calculated stress fields show that their distribution is controlled significantly by the strike of the main faults and can be used to derive depths of the faults. The main faults in Shenzhen may range from 4 km to 20 km in the depth. Each layer of the crust is nearly equipressure since the horizontal tectonic stress has small amplitude. It indicates that the main faults in Shenzhen are relatively stable and have no serious impact on planning and construction of the city.

  3. A Novel Small Molecule, LLL12, Inhibits STAT3 Phosphorylation and Activities and Exhibits Potent Growth-Suppressive Activity in Human Cancer Cells

    Li Lin

    2010-01-01

    Full Text Available Constitutive activation of signal transducer and activator of transcription 3 (STAT3 signaling is frequently detected in cancer, promoting its emergence as a promising target for cancer treatment. Inhibiting constitutive STAT3 signaling represents a potential therapeutic approach. We used structure-based design to develop a nonpeptide, cell-permeable, small molecule, termed as LLL12, which targets STAT3. LLL12 was found to inhibit STAT3 phosphorylation (tyrosine 705 and induce apoptosis as indicated by the increases of cleaved caspase-3 and poly (ADP-ribose polymerase in various breast, pancreatic, and glioblastoma cancer cell lines expressing elevated levels of STAT3 phosphorylation. LLL12 could also inhibit STAT3 phosphorylation induced by interleukin-6 in MDA-MB-453 breast cancer cells. The inhibition of STAT3 by LLL12 was confirmed by the inhibition of STAT3 DNA binding activity and STAT3-dependent transcriptional luciferase activity. Downstream targets of STAT3, cyclin D1, Bcl-2, and survivin were also downregulated by LLL12 at both protein and messenger RNA levels. LLL12 is a potent inhibitor of cell viability, with half-maximal inhibitory concentrations values ranging between 0.16 and 3.09 µM, which are lower than the reported JAK2 inhibitor WP1066 and STAT3 inhibitor S3I-201 in six cancer cell lines expressing elevated levels of STAT3 phosphorylation. In addition, LLL12 inhibits colony formation and cell migration and works synergistically with doxorubicin and gemcitabine. Furthermore, LLL12 demonstrated a potent inhibitory activity on breast and glioblastoma tumor growth in a mouse xenograft model. Our results indicate that LLL12 may be a potential therapeutic agent for human cancer cells expressing constitutive STAT3 signaling.

  4. Immunohistochemical expression of the oncogenic molecules active Stat3 and survivin in benign and malignant salivary gland tumors

    Nikitakis, Nikolaos G.; Scheper, Mark A.; Papanicolaou, Vasileios S.; Sklavounou, Alexandra; Sauk, John J.

    2009-01-01

    Objective Signal transducer and activator of transcription 3 (Stat3) and survivin have been shown to exert oncogenic effects in various human neoplasms. The purpose of this study was to evaluate the expression of the tyrosine phosphorylated (active) Stat3 and survivin in various benign and malignant salivary gland tumors (SGTs). Study design Eighty-six SGTs (65 malignant and 21 benign tumors of various histopathologic subtypes) were immunohistochemically stained with anti-survivin or anti-phosphorylated tyrosine-705 (p-tyr) Stat3 antibodies. Immunohistochemical reactivity was graded in a semi-quantitative manner; a combined score of immunohistochemical positivity (0–6) was calculated for each tumor by adding the individual scores for percentage of tumor cells (0–3) and intensity of staining (0–3). Results Survivin was immunohistochemically detected in all studied benign and malignant SGTs; p-tyr Stat3 was also detected in the majority (91%) of SGTs. The average combined scores for survivin and p-tyr Stat3 immunohistochemical expression in the studied malignant SGTs was 4.40 and 3.35, respectively; the corresponding combined scores for survivin and p-tyr Stat3 in the studied benign SGTs were 4.37 and 3.22, respectively. No statistically significant differences (p>0.05) in p-tyr Stat3 or survivin expression were detected between the benign and malignant groups, or among the various examined histopathological subtypes of SGTs. In contrast, normal salivary gland elements in the vicinity of the studied tumors revealed only weak and focal survivin or p-tyr Stat3 immunoreactivity, mainly localized to ductal and mucous cells. Conclusions Our data indicate an almost universal expression of activated Stat3 and survivin in benign and malignant SGTs. Considering the well-established proliferative and anti-apoptotic properties of these molecules and their functional interrelationship, selective targeting techniques against Stat3 and/or survivin may represent promising

  5. Shh and ZRS enhancer colocalisation is specific to the zone of polarising activity.

    Williamson, Iain; Lettice, Laura A; Hill, Robert E; Bickmore, Wendy A

    2016-08-15

    Limb-specific Shh expression is regulated by the (∼1 Mb distant) ZRS enhancer. In the mouse, limb bud-restricted spatiotemporal Shh expression occurs from ∼E10 to E11.5 at the distal posterior margin and is essential for correct autopod formation. Here, we have analysed the higher-order chromatin conformation of Shh in expressing and non-expressing tissues, both by fluorescence in situ hybridisation (FISH) and by chromosome conformation capture (5C). Conventional and super-resolution light microscopy identified significantly elevated frequencies of Shh/ZRS colocalisation only in the Shh-expressing regions of the limb bud, in a conformation consistent with enhancer-promoter loop formation. However, in all tissues and at all developmental stages analysed, Shh-ZRS spatial distances were still consistently shorter than those to a neural enhancer located between Shh and ZRS in the genome. 5C identified a topologically associating domain (TAD) over the Shh/ZRS genomic region and enriched interactions between Shh and ZRS throughout E11.5 embryos. Shh/ZRS colocalisation, therefore, correlates with the spatiotemporal domain of limb bud-specific Shh expression, but close Shh and ZRS proximity in the nucleus occurs regardless of whether the gene or enhancer is active. We suggest that this constrained chromatin configuration optimises the opportunity for the active enhancer to locate and instigate the expression of Shh.

  6. Active flexural-slip faulting: A study from the Pamir-Tian Shan convergent zone, NW China

    Li, Tao; Chen, Jie; Thompson, Jessica A.; Burbank, Douglas W.; Yang, Xiaodong

    2015-06-01

    The flexural-slip fault (FSF), a type of secondary fault generated by bed-parallel slip, occurs commonly and plays an important role in accommodating fold growth. Although the kinematics and mechanics of FSFs are well studied, relatively few field observations or geometric models explore its geomorphic expression. In the Pamir-Tian Shan convergent zone, NW China, suites of well-preserved FSF scarps displace fluvial terraces in the Mingyaole and Wulagen folds. Integrating interpretations of Google Earth images, detailed geologic and geomorphic mapping, and differential GPS measurements of terrace surfaces, we summarize geomorphic features that typify these faults and create kinematic models of active flexural-slip faulting. Our study indicates the following: (i) FSF scarps commonly occur near synclinal hinges, irrespective of whether (a) the dip direction of beds on either side of the hinge is unidirectional or in opposite directions, (b) the hinge is migrating or fixed, or (c) the hinge shape is narrow and angular or wide and curved. (ii) Active FSFs are likely to produce higher scarps on steeper beds, whereas lower or no topographic scarps typify gentler beds. (iii) Tilt angles of the terrace surface displaced above FSFs progressively decrease farther away from the hinge, with abrupt changes in slope coinciding with FSF scarps; the changes in tilt angle and scarp height have a predictable geometric relationship. (iv) Active FSFs can accommodate a significant fraction of total slip and play a significant role in folding deformation. (v) Active FSFs may be used to assess seismic hazards associated with active folds and associated blind thrusts.

  7. Cytometric analysis of surface molecules of leucocytes and phagocytic activity of granulocytes and monocytes/macrophages in cows with pyometra.

    Brodzki, P; Kostro, K; Brodzki, A; Niemczuk, K; Lisiecka, U

    2014-10-01

    Pyometra is a serious problem in dairy cow herds, causing large economic losses due to infertility. The development of pyometra depends mainly on the immunological status of the cow. The aim of the study was a comparative evaluation of selected indicators involving non-specific and specific immunity in cows with pyometra and in cows without inflammation of the uterus. The study was performed in 20 cows, which were divided into two groups: pyometra group and healthy group, each comprising 10 cows, based on the results of cytological and ultrasonographic tests. A flow cytometric analysis was performed for the surface molecules CD4, CD8, CD14, CD21, CD25 and CD4(+) CD25(+) on leucocytes, and the phagocytic activity was determined from granulocytes and monocytes/macrophages in the peripheral blood and uterine washings, respectively. It was demonstrated that the percentage of phagocytic granulocytes and monocytes/macrophages in both the peripheral blood and uterine washings was significantly lower in cows with pyometra compared with the healthy group (p < 0.001). Significantly (p ≤ 0.001) lower percentage of CD4(+) , CD14(+) , CD25(+) and CD4(+) CD25(+) phenotype leucocytes was also observed in the peripheral blood of cows from the pyometra group, along with a significantly higher (p < 0.001) percentage of CD8(+) and CD21(+) lymphocytes as compared to the healthy group. The results of work indicate that disfunction of cell immunity coexisting with pyometra may be caused by a bacterial infection and the presence of blocking agents (IL-10), released by the increasing number of CD8(+) lymphocytes what leads to the advanced inflammation of uterus.

  8. Joint molecule resolution requires the redundant activities of MUS-81 and XPF-1 during Caenorhabditis elegans meiosis.

    Nigel J O'Neil

    Full Text Available The generation and resolution of joint molecule recombination intermediates is required to ensure bipolar chromosome segregation during meiosis. During wild type meiosis in Caenorhabditis elegans, SPO-11-generated double stranded breaks are resolved to generate a single crossover per bivalent and the remaining recombination intermediates are resolved as noncrossovers. We discovered that early recombination intermediates are limited by the C. elegans BLM ortholog, HIM-6, and in the absence of HIM-6 by the structure specific endonuclease MUS-81. In the absence of both MUS-81 and HIM-6, recombination intermediates persist, leading to chromosome breakage at diakinesis and inviable embryos. MUS-81 has an additional role in resolving late recombination intermediates in C. elegans. mus-81 mutants exhibited reduced crossover recombination frequencies suggesting that MUS-81 is required to generate a subset of meiotic crossovers. Similarly, the Mus81-related endonuclease XPF-1 is also required for a subset of meiotic crossovers. Although C. elegans gen-1 mutants have no detectable meiotic defect either alone or in combination with him-6, mus-81 or xpf-1 mutations, mus-81;xpf-1 double mutants are synthetic lethal. While mus-81;xpf-1 double mutants are proficient for the processing of early recombination intermediates, they exhibit defects in the post-pachytene chromosome reorganization and the asymmetric disassembly of the synaptonemal complex, presumably triggered by crossovers or crossover precursors. Consistent with a defect in resolving late recombination intermediates, mus-81; xpf-1 diakinetic bivalents are aberrant with fine DNA bridges visible between two distinct DAPI staining bodies. We were able to suppress the aberrant bivalent phenotype by microinjection of activated human GEN1 protein, which can cleave Holliday junctions, suggesting that the DNA bridges in mus-81; xpf-1 diakinetic oocytes are unresolved Holliday junctions. We propose that the

  9. Recent host range expansion of canine distemper virus and variation in its receptor, the signaling lymphocyte activation molecule, in carnivores.

    Ohishi, Kazue; Suzuki, Rintaro; Maeda, Taro; Tsuda, Miwako; Abe, Erika; Yoshida, Takao; Endo, Yasuyuki; Okamura, Maki; Nagamine, Takashi; Yamamoto, Hanae; Ueda, Miya; Maruyama, Tadashi

    2014-07-01

    The signaling lymphocyte activation molecule (SLAM) is a receptor for morbilliviruses. To understand the recent host range expansion of canine distemper virus (CDV) in carnivores, we determined the nucleotide sequences of SLAMs of various carnivores and generated three-dimensional homology SLAM models. Thirty-four amino acid residues were found for the candidates binding to CDV on the interface of the carnivore SLAMs. SLAM of the domestic dog (Canis lupus familiaris) were similar to those of other members of the suborder Caniformia, indicating that the animals in this group have similar sensitivity to dog CDV. However, they were different at nine positions from those of felids. Among the nine residues, four of domestic cat (Felis catus) SLAM (72, 76, 82, and 129) and three of lion (Panthera leo persica) SLAM (72, 82, and 129) were associated with charge alterations, suggesting that the felid interfaces have lower affinities to dog CDV. Only the residue at 76 was different between domestic cat and lion SLAM interfaces. The domestic cat SLAM had threonine at 76, whereas the lion SLAM had arginine, a positively charged residue like that of the dog SLAM. The cat SLAM with threonine is likely to have lower affinity to CDV-H and to confer higher resistance against dog CDV. Thus, the four residues (72, 76, 82, and 129) on carnivore SLAMs are important for the determination of affinity and sensitivity with CDV. Additionally, the CDV-H protein of felid strains had a substitution of histidine for tyrosine at 549 of dog CDV-H and may have higher affinity to lion SLAM. Three-dimensional model construction is a new risk assessment method of morbillivirus infectivity. Because the method is applicable to animals that have no information about virus infection, it is especially useful for morbillivirus risk assessment and wildlife conservation.

  10. Could both vitamin D and geomagnetic activity impact serum levels of soluble cell adhesion molecules in young men?

    Bleizgys, Andrius; Šapoka, Virginijus

    2016-07-01

    Vitamin D might have a role in diminishing endothelial dysfunction (ED). The initial aim was to test the hypothesis of reciprocity between levels of 25-hydroxyvitamin D (25(OH)D) and levels of soluble endothelial cell adhesion molecules (CAMs) that could serve as biomarkers of ED. Randomly selected men of age 20-39 were examined at February or March (cold season) and reexamined at August or September (warm season). Some lifestyle and anthropometrical data were recorded. Laboratory measurements, including those for serum levels of soluble CAMs—sICAM-1, sVCAM-1, sE-selectin and sP-selectin—were also performed. As some of the results were rather unexpected, indices of geomagnetic activity (GMA), obtained from the online database, were included in further analysis as a confounder. In 2012-2013, 130 men were examined in cold season, and 125 of them were reexamined in warm season. 25(OH)D levels were found to be significantly negatively associated with sVCAM-1 levels ( β = -0.15, p = 0.043 in warm season; β = -0.19, p = 0.007 for changes). Levels of sVCAM-1 and sICAM-1 from the same seasons were notably different between years and have changed in an opposite manner. Soluble P-selectin levels were higher at warm season in both years. GMA was positively associated with sVCAM-1 ( β = 0.17, p = 0.039 in cold season; β = 0.22, p = 0.002 for changes) and negatively with sICAM-1 ( β = -0.30. p Vitamin D might play a role in diminishing sVCAM-1 levels. Levels of sVCAM-1 and sICAM-1 were associated with the GMA; this implies a need for further research.

  11. Increase in electron transfer activity in photosystem Ⅱ of spinach thylakoids caused by conversion of phosphatidyl-glycerol to phosphatidic acid molecules

    WU Feng; YANG Zhenle; LI Liangbi; KUANG Tingyun

    2003-01-01

    The techniques of oxygen electrode polarography, sodium dodecyl sulfate-polyacryamide gel electrophoresis (SDS-PAGE) and thin layer chromatography (TLC) were employed to investigate the effect of phospholipase D treatment on physiological function of spinach thylakoids. It was shown that the phospholipase D treatment on thylakoid resulted in the degradation of phosphatidylglycerol (PG) and occurrence of phosphatidic acid (PA). The changes of PG to PA molecules caused an increase in oxygen evolution in photosystem Ⅱ (PSⅡ), which was accompanied by an uncoupling effect on thylakoid membrane. It was revealed that the head-groups of PG molecules play an important role in the maintenance of the appropriate physiological activity of thylakoid membrane.

  12. Effect of alteration of translation error rate on enzyme microheterogeneity as assessed by variation in single molecule electrophoretic mobility and catalytic activity.

    Nichols, Ellert R; Shadabi, Elnaz; Craig, Douglas B

    2009-06-01

    The role of translation error for Escherichia coli individual beta-galactosidase molecule catalytic and electrophoretic heterogeneity was investigated using CE-LIF. An E. coli rpsL mutant with a hyperaccurate translation phenotype produced enzyme molecules that exhibited significantly less catalytic heterogeneity but no reduction of electrophoretic heterogeneity. Enzyme expressed with streptomycin-induced translation error had increased thermolability, lower activity, and no significant change to catalytic or electrophoretic heterogeneity. Modeling of the electrophoretic behaviour of beta-galactosidase suggested that variation of the hydrodynamic radius may be the most significant contributor to electrophoretic heterogeneity.

  13. In vitro antibacterial activity of ceftobiprole against clinical isolates from French teaching hospitals: proposition of zone diameter breakpoints.

    Lascols, C; Legrand, P; Mérens, A; Leclercq, R; Muller-Serieys, C; Drugeon, H B; Kitzis, M D; Reverdy, M E; Roussel-Delvallez, M; Moubareck, C; Brémont, S; Miara, A; Gjoklaj, M; Soussy, C-J

    2011-03-01

    The aims of this study were to determine the in vitro activity profile of ceftobiprole, a pyrrolidinone cephalosporin, against a large number of bacterial pathogens and to propose zone diameter breakpoints for clinical categorisation according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) minimum inhibitory concentration (MIC) breakpoints. MICs of ceftobiprole were determined by broth microdilution against 1548 clinical isolates collected in eight French hospitals. Disk diffusion testing was performed using 30 μg disks according to the method of the Comité de l'Antibiogramme de la Société Française de Microbiologie (CA-SFM). The in vitro activity of ceftobiprole, expressed by MIC(50/90) (MICs for 50% and 90% of the organisms, respectively) (mg/L), was as follows: meticillin-susceptible Staphylococcus aureus, 0.25/0.5; meticillin-resistant S. aureus (MRSA), 1/2; meticillin-susceptible coagulase-negative staphylococci (CoNS), 0.12/0.5; meticillin-resistant CoNS, 1/2; penicillin-susceptible Streptococcus pneumoniae, ≤ 0.008/0.03; penicillin-resistant S. pneumoniae, 0.12/0.5; viridans group streptococci, 0.03/0.12; β-haemolytic streptococci, ≤ 0.008/0.016; Enterococcus faecalis, 0.25/1; Enterococcus faecium, 64/128; Enterobacteriaceae, 0.06/32; Pseudomonas aeruginosa, 4/16; Acinetobacter baumannii, 0.5/64; Haemophilus influenzae, 0.03/0.12; and Moraxella catarrhalis, 0.25/0.5. According to the regression curve, zone diameter breakpoints could be 28, 26, 24 and 22 mm for MICs of 0.5, 1, 2 and 4 mg/L respectively. In conclusion, this study confirms the potent in vitro activity of ceftobiprole against many Gram-positive bacteria, including MRSA but not E. faecium, whilst maintaining a Gram-negative spectrum similar to the advanced-generation cephalosporins such as cefepime. Thus ceftobiprole appears to be well suited for the empirical treatment of a variety of healthcare-associated infections.

  14. Enumerating molecules.

    Visco, Donald Patrick, Jr. (, . Tennessee Technological University, Cookeville, TN); Faulon, Jean-Loup Michel; Roe, Diana C.

    2004-04-01

    This report is a comprehensive review of the field of molecular enumeration from early isomer counting theories to evolutionary algorithms that design molecules in silico. The core of the review is a detail account on how molecules are counted, enumerated, and sampled. The practical applications of molecular enumeration are also reviewed for chemical information, structure elucidation, molecular design, and combinatorial library design purposes. This review is to appear as a chapter in Reviews in Computational Chemistry volume 21 edited by Kenny B. Lipkowitz.

  15. Activation of rape (Brassica napus L. embryo during seed germination. V. The first zones of ultrastructural changes and their expansion

    Mieczysław Karaś

    2014-01-01

    Full Text Available In the germinating rape embryo the columella and basal part of hypocotyl undergo earliest activation. Its first ultrastructural symptom is the appearance of numerous ER vesicles after 3-6 h of seed swelling. Their number is the highest in the external layers of the columella and decreases in basipetal direction. Dermatogen cells in the basal zone of the hypocotyl contain the greatest amount of ER structures, whereas decreasing amounts are found in both directions along the embryo axis and centripetally. Further changes in the ER spread in a similar order. The vesicles merge and form a tubular and plate-like ER. Then, they disappear and are replaced by tubular and vesicular forms. The changes in the ER are gradually followed by ultrastructural symptoms of activation of mitochondria, plastids and dictyosomes. The highest number of ER structures and other organelles accumulate in root cells shortly before piercing of the seed coat. After germination their amount decreases and remains almost stable.

  16. The influence of galvanic currents and voltage on the proliferation activity of lymphocytes and expression of cell surface molecules.

    Podzimek, S; Hána, K; Miksovský, M; Pousek, L; Matucha, P; Meloun, M; Procházková, J

    2008-01-01

    Release of metal ions from dental metal fillings supported by galvanism can cause local or general pathological problems in sensitive and genetically susceptible individuals. We aimed to investigate in vitro lymphocyte responses and expression of surface molecules influenced by galvanic currents and voltage. Human peripheral blood lymphocytes were influenced by galvanic currents and voltages and lymphocyte proliferation was measured. Control samples were not exposed to the influence of galvanism. We also studied the expression of surface molecules by the FACS analysis. A 15-h and shorter influence of almost all tested currents and voltages caused a significant decrease in lymphocyte proliferation and the 15-h influence of 20 microA currents significantly increased expression of surface molecules CD 19, 11a/18, 19/69 and 19/95. An influence of 10 and 3 microA currents led to a significant decrease in the expression of surface molecules CD 3, 11a/18, 3/69 and 3/95 and to a significant increase in CD 19 expression. An 80 mV voltage influence led to a significant decrease in the expression of surface molecules CD 3, 11a/18, 3/69, 3/95, 19/69 and 19/95, and 200 and 300 mV voltages significantly decreased the expression of surface molecules CD 3, 19, 11a/18, 3/95 and 19/95 and significantly increased CD 19/69 expression. A long-lasting influence of galvanism can, in sensitive and genetically susceptible individuals, influence lymphocyte proliferation and surface molecule expression. The threshold for pathological values of 5 microA for galvanic currents and 100 mV for galvanic voltage was confirmed.

  17. Activation of type 1 cannabinoid receptor (CB1R promotes neurogenesis in murine subventricular zone cell cultures.

    Sara Xapelli

    Full Text Available The endocannabinoid system has been implicated in the modulation of adult neurogenesis. Here, we describe the effect of type 1 cannabinoid receptor (CB1R activation on self-renewal, proliferation and neuronal differentiation in mouse neonatal subventricular zone (SVZ stem/progenitor cell cultures. Expression of CB1R was detected in SVZ-derived immature cells (Nestin-positive, neurons and astrocytes. Stimulation of the CB1R by (R-(+-Methanandamide (R-m-AEA increased self-renewal of SVZ cells, as assessed by counting the number of secondary neurospheres and the number of Sox2+/+ cell pairs, an effect blocked by Notch pathway inhibition. Moreover, R-m-AEA treatment for 48 h, increased proliferation as assessed by BrdU incorporation assay, an effect mediated by activation of MAPK-ERK and AKT pathways. Surprisingly, stimulation of CB1R by R-m-AEA also promoted neuronal differentiation (without affecting glial differentiation, at 7 days, as shown by counting the number of NeuN-positive neurons in the cultures. Moreover, by monitoring intracellular calcium concentrations ([Ca(2+]i in single cells following KCl and histamine stimuli, a method that allows the functional evaluation of neuronal differentiation, we observed an increase in neuronal-like cells. This proneurogenic effect was blocked when SVZ cells were co-incubated with R-m-AEA and the CB1R antagonist AM 251, for 7 days, thus indicating that this effect involves CB1R activation. In accordance with an effect on neuronal differentiation and maturation, R-m-AEA also increased neurite growth, as evaluated by quantifying and measuring the number of MAP2-positive processes. Taken together, these results demonstrate that CB1R activation induces proliferation, self-renewal and neuronal differentiation from mouse neonatal SVZ cell cultures.

  18. Carbon Dioxide Activation by Scandium Atoms and Scandium Monoxide Molecules: Formation and Spectroscopic Characterization of ScCO3 and OCScCO3 in Solid Neon.

    Zhang, Qingnan; Qu, Hui; Chen, Mohua; Zhou, Mingfei

    2016-01-28

    The reactions of carbon dioxide with scandium monoxide molecules and scandium atoms are investigated using matrix isolation infrared spectroscopy in solid neon. The species formed are identified by the effects of isotopic substitution on their infrared spectra as well as density functional calculations. The results show that the ground state ScO molecule reacts with carbon dioxide to form the carbonate complex ScCO3 spontaneously on annealing. The ground state Sc atom reacts with two carbon dioxide molecules to give the carbonate carbonyl complex OCScCO3 via the previously reported OScCO insertion intermediate on annealing. The observation of these spontaneous reactions is consistent with theoretical predictions that both the Sc + 2CO2 → OCScCO3 and ScO + CO2 → ScCO3 reactions are thermodynamically exothermic and are kinetically facile, requiring little or no activation energy.

  19. Pollution from organic contaminants in Greek marine areas, receiving anthropogenic pressures from intense activities in the coastal zone

    Hatzianestis, Ioannis

    2014-05-01

    Polycyclic aromatic hydrocarbons (PAHs) are widespread pollutants in marine sediments, receiving the pressures from various anthropogenic activities in the coastal zone. Due to their mutagenic and carcinogenic behaviour, PAHs are classified as priority contaminants to be monitored in environmental quality control schemes. The purpose of this study was to determine the levels of PAHs in coastal areas of Greece directly influenced from the operation of major industrial units in the coastal zone, investigate their sources and evaluate their potential toxicity by comparison against effect - based sediment quality guidelines. Thirty two surface sediment samples were collected from three areas of the Hellenic coastline: a) Antikyra bay in Korinthiakos gulf, influenced from the operation of an alumina and aluminium production plant b) Larymna bay in Noth Evoikos gulf, influenced from the operation of a nickel production plant and c) Aliveri bay in South Evoikos Gulf, influenced from a cement production plant. In all the areas studied, aquaculture and fishing activities have been also developed in the coastal zone. PAH concentrations were determined by GC-MS, after soxhlet extraction and fractionation by silica column chromatography. PAH sources and origin were investigated by applying several isomeric ratio diagnostic criteria. The mean quotient Effect- Range Median (m-ERM) was used to evaluate the potential of adverse effects posed to benthic organisms. Three m-ERM-q values were used to differentiate the probability of observing toxicity and classify sites into four categories: sediments with m-ERM1.5 have the highest probability (76%) of toxicity. Extremely high PAH concentrations more than 100,000 ng/g were found in the close vicinity of the alumina production plant in Antikyra bay. High levels of PAHs up to 22,000 ng/g were also found in Aliveri bay, whereas lowest values, but still indicating significant pollution, were measured close to the nickel production plant

  20. Ventilation rates and activity levels of juvenile jumbo squid under metabolic suppression in the oxygen minimum zone.

    Trübenbach, Katja; Pegado, Maria R; Seibel, Brad A; Rosa, Rui

    2013-02-01

    The Humboldt (jumbo) squid, Dosidicus gigas, is a part-time resident of the permanent oxygen minimum zone (OMZ) in the Eastern Tropical Pacific and, thereby, it encounters oxygen levels below its critical oxygen partial pressure. To better understand the ventilatory mechanisms that accompany the process of metabolic suppression in these top oceanic predators, we exposed juvenile D. gigas to the oxygen levels found in the OMZ (1% O(2), 1 kPa, 10 °C) and measured metabolic rate, activity cycling patterns, swimming mode, escape jet (burst) frequency, mantle contraction frequency and strength, stroke volume and oxygen extraction efficiency. In normoxia, metabolic rate varied between 14 and 29 μmol O(2) g(-1) wet mass h(-1), depending on the level of activity. The mantle contraction frequency and strength were linearly correlated and increased significantly with activity level. Additionally, an increase in stroke volume and ventilatory volume per minute was observed, followed by a mantle hyperinflation process during high activity periods. Squid metabolic rate dropped more than 75% during exposure to hypoxia. Maximum metabolic rate was not achieved under such conditions and the metabolic scope was significantly decreased. Hypoxia changed the relationship between mantle contraction strength and frequency from linear to polynomial with increasing activity, indicating that, under hypoxic conditions, the jumbo squid primarily increases the strength of mantle contraction and does not regulate its frequency. Under hypoxia, jumbo squid also showed a larger inflation period (reduced contraction frequency) and decreased relaxed mantle diameter (shortened diffusion pathway), which optimize oxygen extraction efficiency (up to 82%/34%, without/with consideration of 60% potential skin respiration). Additionally, they breathe 'deeply', with more powerful contractions and enhanced stroke volume. This deep-breathing behavior allows them to display a stable ventilatory volume per

  1. A strategy for the incorporation of water molecules present in a ligand binding site into a three-dimensional quantitative structure--activity relationship analysis.

    Pastor, M; Cruciani, G; Watson, K A

    1997-12-05

    Water present in a ligand binding site of a protein has been recognized to play a major role in ligand-protein interactions. To date, rational drug design techniques do not usually incorporate the effect of these water molecules into the design strategy. This work represents a new strategy for including water molecules into a three-dimensional quantitative structure-activity relationship analysis using a set of glucose analogue inhibitors of glycogen phosphorylase (GP). In this series, the structures of the ligand-enzyme complexes have been solved by X-ray crystallography, and the positions of the ligands and the water molecules at the ligand binding site are known. For the structure-activity analysis, some water molecules adjacent to the ligands were included into an assembly which encompasses both the inhibitor and the water involved in the ligand-enzyme interaction. The mobility of some water molecules at the ligand binding site of GP gives rise to differences in the ligand-water assembly which have been accounted for using a simulation study involving force-field energy calculations. The assembly of ligand plus water was used in a GRID/GOLPE analysis, and the models obtained compare favorably with equivalent models when water was excluded. Both models were analyzed in detail and compared with the crystallographic structures of the ligand-enzyme complexes in order to evaluate their ability to reproduce the experimental observations. The results demonstrate that incorporation of water molecules into the analysis improves the predictive ability of the models and makes them easier to interpret. The information obtained from interpretation of the models is in good agreement with the conclusions derived from the structural analysis of the complexes and offers valuable insights into new characteristics of the ligands which may be exploited for the design of more potent inhibitors.

  2. Molecules in galaxies

    Omont, Alain

    2007-01-01

    The main achievements, current developments and prospects of molecular studies in external galaxies are reviewed. They are put in the context of the results of several decades of studies of molecules in local interstellar medium, their chemistry and their importance for star formation. CO observations have revealed the gross structure of molecular gas in galaxies. Together with other molecules, they are among the best tracers of star formation at galactic scales. Our knowledge about molecular abundances in various local galactic environments is progressing. They trace physical conditions and metallicity, and they are closely related to dust processes and large aromatic molecules. Major recent developments include mega-masers, and molecules in Active Galactic Nuclei; millimetre emission of molecules at very high redshift; and infrared H2 emission as tracer of warm molecular gas, shocks and photodissociation regions. The advent of sensitive giant interferometers from the centimetre to sub-millimetre range, espe...

  3. Domain movements of the enhancer-dependent sigma factor drive DNA delivery into the RNA polymerase active site: insights from single molecule studies.

    Sharma, Amit; Leach, Robert N; Gell, Christopher; Zhang, Nan; Burrows, Patricia C; Shepherd, Dale A; Wigneshweraraj, Sivaramesh; Smith, David Alastair; Zhang, Xiaodong; Buck, Martin; Stockley, Peter G; Tuma, Roman

    2014-04-01

    Recognition of bacterial promoters is regulated by two distinct classes of sequence-specific sigma factors, σ(70) or σ(54), that differ both in their primary sequence and in the requirement of the latter for activation via enhancer-bound upstream activators. The σ(54) version controls gene expression in response to stress, often mediating pathogenicity. Its activator proteins are members of the AAA+ superfamily and use adenosine triphosphate (ATP) hydrolysis to remodel initially auto-inhibited holoenzyme promoter complexes. We have mapped this remodeling using single-molecule fluorescence spectroscopy. Initial remodeling is nucleotide-independent and driven by binding both ssDNA during promoter melting and activator. However, DNA loading into the RNA polymerase active site depends on co-operative ATP hydrolysis by the activator. Although the coupled promoter recognition and melting steps may be conserved between σ(70) and σ(54), the domain movements of the latter have evolved to require an activator ATPase.

  4. Introducing Bond-Line Organic Structures in High School Biology: An Activity that Incorporates Pleasant-Smelling Molecules

    Rios, Andro C.; French, Gerald

    2011-01-01

    Chemical education occurs in settings other than just the chemistry classroom. High school biology courses are frequently where students are introduced to organic molecules and their importance to cellular chemistry. However, structural representations are often intimidating because students have not been introduced to the language. As part of a…

  5. A high affinity RIM-binding protein/Aplip1 interaction prevents the formation of ectopic axonal active zones

    Siebert, Matthias; Böhme, Mathias A; Driller, Jan H; Babikir, Husam; Mampell, Malou M; Rey, Ulises; Ramesh, Niraja; Matkovic, Tanja; Holton, Nicole; Reddy-Alla, Suneel; Göttfert, Fabian; Kamin, Dirk; Quentin, Christine; Klinedinst, Susan; Andlauer, Till FM; Hell, Stefan W; Collins, Catherine A; Wahl, Markus C; Loll, Bernhard; Sigrist, Stephan J

    2015-01-01

    Synaptic vesicles (SVs) fuse at active zones (AZs) covered by a protein scaffold, at Drosophila synapses comprised of ELKS family member Bruchpilot (BRP) and RIM-binding protein (RBP). We here demonstrate axonal co-transport of BRP and RBP using intravital live imaging, with both proteins co-accumulating in axonal aggregates of several transport mutants. RBP, via its C-terminal Src-homology 3 (SH3) domains, binds Aplip1/JIP1, a transport adaptor involved in kinesin-dependent SV transport. We show in atomic detail that RBP C-terminal SH3 domains bind a proline-rich (PxxP) motif of Aplip1/JIP1 with submicromolar affinity. Pointmutating this PxxP motif provoked formation of ectopic AZ-like structures at axonal membranes. Direct interactions between AZ proteins and transport adaptors seem to provide complex avidity and shield synaptic interaction surfaces of pre-assembled scaffold protein transport complexes, thus, favouring physiological synaptic AZ assembly over premature assembly at axonal membranes. DOI: http://dx.doi.org/10.7554/eLife.06935.001 PMID:26274777

  6. Compound 13, an α1-selective small molecule activator of AMPK, inhibits Helicobacter pylori-induced oxidative stresses and gastric epithelial cell apoptosis

    Zhao, Hangyong; Zhu, Huanghuang; Lin, Zhou; Lin, Gang; Lv, Guoqiang, E-mail: lvguoqiangwuxivip@163.com

    2015-08-07

    Half of the world's population experiences Helicobacter pylori (H. pylori) infection, which is a main cause of gastritis, duodenal and gastric ulcer, and gastric cancers. In the current study, we investigated the potential role of compound 13 (C13), a novel α1-selective small molecule activator of AMP-activated protein kinase (AMPK), against H. pylori-induced cytotoxicity in cultured gastric epithelial cells (GECs). We found that C13 induced significant AMPK activation, evidenced by phosphorylation of AMPKα1 and ACC (acetyl-CoA carboxylase), in both primary and transformed GECs. Treatment of C13 inhibited H. pylori-induced GEC apoptosis. AMPK activation was required for C13-mediated GEC protection. Inhibition of AMPK kinase activity by the AMPK inhibitor Compound C, or silencing AMPKα1 expression by targeted-shRNAs, alleviated C13-induced GEC protective activities against H. pylori. Significantly, C13 inhibited H. pylori-induced reactive oxygen species (ROS) production in GECs. C13 induced AMPK-dependent expression of anti-oxidant gene heme oxygenase (HO-1) in GECs. Zinc protoporphyrin (ZnPP) and tin protoporphyrin (SnPP), two HO-1 inhibitors, not only suppressed C13-mediated ROS scavenging activity, but also alleviated its activity in GECs against H. pylori. Together, these results indicate that C13 inhibits H. pylori-induced ROS production and GEC apoptosis through activating AMPK–HO–1 signaling. - Highlights: • We synthesized compound 13 (C13), a α1-selective small molecule AMPK activator. • C13-induced AMPK activation requires α1 subunit in gastric epithelial cells (GECs). • C13 enhances Helicobacter pylori-induced pro-survival AMPK activation to inhibit GEC apoptosis. • C13 inhibits H. pylori-induced reactive oxygen species (ROS) production in GECs. • AMPK-heme oxygenase (HO-1) activation is required for C13-mediated anti-oxidant activity.

  7. Specificity and mechanism of action of alpha-helical membrane-active peptides interacting with model and biological membranes by single-molecule force spectroscopy.

    Sun, Shiyu; Zhao, Guangxu; Huang, Yibing; Cai, Mingjun; Shan, Yuping; Wang, Hongda; Chen, Yuxin

    2016-07-01

    In this study, to systematically investigate the targeting specificity of membrane-active peptides on different types of cell membranes, we evaluated the effects of peptides on different large unilamellar vesicles mimicking prokaryotic, normal eukaryotic, and cancer cell membranes by single-molecule force spectroscopy and spectrum technology. We revealed that cationic membrane-active peptides can exclusively target negatively charged prokaryotic and cancer cell model membranes rather than normal eukaryotic cell model membranes. Using Acholeplasma laidlawii, 3T3-L1, and HeLa cells to represent prokaryotic cells, normal eukaryotic cells, and cancer cells in atomic force microscopy experiments, respectively, we further studied that the single-molecule targeting interaction between peptides and biological membranes. Antimicrobial and anticancer activities of peptides exhibited strong correlations with the interaction probability determined by single-molecule force spectroscopy, which illustrates strong correlations of peptide biological activities and peptide hydrophobicity and charge. Peptide specificity significantly depends on the lipid compositions of different cell membranes, which validates the de novo design of peptide therapeutics against bacteria and cancers.

  8. Crustal Structure Across the Okavango Rift Zone, Botswana: Initial Results From the PRIDE-SEISORZ Active-Source Seismic Profile

    Canales, J. P.; Moffat, L.; Lizarralde, D.; Laletsang, K.; Harder, S. H.; Kaip, G.; Modisi, M.

    2015-12-01

    The PRIDE project aims to understand the processes of continental rift initiation and evolution by analyzing along-axis trends in the southern portion of the East Africa Rift System, from Botswana through Zambia and Malawi. The SEISORZ active-source seismic component of PRIDE focused on the Okavango Rift Zone (ORZ) in northwestern Botswana, with the main goal of imaging the crustal structure across the ORZ. This will allow us to estimate total crustal extension, determine the pattern and amount of thinning, assess the possible presence of melt within the rift zone, and assess the contrasts in crustal blocks across the rift, which closely follows the trend of a fold belt. In November 2014 we conducted a crustal-scale, 450-km-long seismic refraction/wide-angle reflection profile consisting of 19 sources (shots in 30-m-deep boreholes) spaced ~25 km apart from each other, and 900 receivers (IRIS/PASSCAL "Texan" dataloggers and 4.5Hz geophones) with ~500 m spacing. From NW to SE, the profile crosses several tectonic domains: the Congo craton, the Damara metamorphic belt and the Ghanzi-Chobe fold belt where the axis of the ORZ is located, and continues into the Kalahari craton. The record sections display clear crustal refraction (Pg) and wide-angle Moho reflection (PmP) phases for all 17 of the good-quality shots, and a mantle refraction arrival (Pn), with the Pg-PmP-Pn triplication appearing at 175 km offset. There are distinct changes in the traveltime and amplitude of these phases along the transect, and on either side of the axis, that seem to correlate with sharp transitions across tectonic terrains. Initial modeling suggests: (1) the presence of a sedimentary half-graben structure at the rift axis beneath the Okavango delta, bounded to the SE by the Kunyere-Thamalakane fault system; (2) faster crustal Vp in the domains to the NW of the ORZ; and (3) thicker crust (45-50 km) at both ends of the profile within the Congo and Kalahari craton domains than at the ORZ and

  9. Soil Moisture Active Passive (SMAP) Mission Level 4 Surface and Root Zone Soil Moisture (L4_SM) Product Specification Document

    Reichle, Rolf H.; Ardizzone, Joseph V.; Kim, Gi-Kong; Lucchesi, Robert A.; Smith, Edmond B.; Weiss, Barry H.

    2015-01-01

    This is the Product Specification Document (PSD) for Level 4 Surface and Root Zone Soil Moisture (L4_SM) data for the Science Data System (SDS) of the Soil Moisture Active Passive (SMAP) project. The L4_SM data product provides estimates of land surface conditions based on the assimilation of SMAP observations into a customized version of the NASA Goddard Earth Observing System, Version 5 (GEOS-5) land data assimilation system (LDAS). This document applies to any standard L4_SM data product generated by the SMAP Project. The Soil Moisture Active Passive (SMAP) mission will enhance the accuracy and the resolution of space-based measurements of terrestrial soil moisture and freeze-thaw state. SMAP data products will have a noteworthy impact on multiple relevant and current Earth Science endeavors. These include: Understanding of the processes that link the terrestrial water, the energy and the carbon cycles, Estimations of global water and energy fluxes over the land surfaces, Quantification of the net carbon flux in boreal landscapes Forecast skill of both weather and climate, Predictions and monitoring of natural disasters including floods, landslides and droughts, and Predictions of agricultural productivity. To provide these data, the SMAP mission will deploy a satellite observatory in a near polar, sun synchronous orbit. The observatory will house an L-band radiometer that operates at 1.40 GHz and an L-band radar that operates at 1.26 GHz. The instruments will share a rotating reflector antenna with a 6 meter aperture that scans over a 1000 km swath.

  10. Activity-dependent endogenous taurine release facilitates excitatory neurotransmission in the neocortical marginal zone of neonatal rats

    Taizhe eQian

    2014-02-01

    Full Text Available In the developing cerebral cortex, the marginal zone (MZ, consisting of early-generated neurons such as Cajal-Retzius cells, plays an important role in cell migration and lamination. There is accumulating evidence of widespread excitatory neurotransmission mediated by γ-aminobutyric acid (GABA in the MZ. Cajal-Retzius cells express not only GABAA receptors but also α2/β subunits of glycine receptors, and exhibit glycine receptor-mediated depolarization due to high [Cl−]i. However, the physiological roles of glycine receptors and their endogenous agonists during neurotransmission in the MZ are yet to be elucidated. To address this question, we performed optical imaging from the MZ using the voltage-sensitive dye JPW1114 on tangential neocortical slices of neonatal rats. A single electrical stimulus evoked an action-potential-dependent optical signal that spread radially over the MZ. The amplitude of the signal was not affected by glutamate receptor blockers, but was suppressed by either GABAA or glycine receptor antagonists. Combined application of both antagonists nearly abolished the signal. Inhibition of Na+, K+-2Cl− cotransporter by 20 µM bumetanide reduced the signal, indicating that this transporter contributes to excitation. Analysis of the interstitial fluid obtained by microdialysis from tangential neocortical slices with high-performance liquid chromatography revealed that GABA and taurine, but not glycine or glutamate, were released in the MZ in response to the electrical stimulation. The ambient release of taurine was reduced by the addition of a voltage-sensitive Na+ channel blocker. Immunohistochemistry and immunoelectron microscopy indicated that taurine was stored both in Cajal-Retzius and non-Cajal-Retzius cells in the MZ, but was not localized in presynaptic structures. Our results suggest that activity-dependent non-synaptic release of endogenous taurine facilitates excitatory neurotransmission through activation of

  11. Antimicrobial activities of endophytic fungi obtained from the arid zone invasive plant Opuntia dillenii and the isolation of equisetin, from endophytic Fusarium sp.

    Ratnaweera, Pamoda B.; de Silva, E. Dilip; Williams, David E.; Andersen, Raymond J

    2015-01-01

    Background Opuntia dillenii is an invasive plant well established in the harsh South-Eastern arid zone of Sri Lanka. Evidence suggests it is likely that the endophytic fungal populations of O. dillenii assist the host in overcoming biotic and abiotic stress by producing biologically active metabolites. With this in mind there is potential to discover novel natural products with useful biological activities from this hitherto poorly investigated source. Consequently, an investigation of the an...

  12. Immunological network activation by low-dose rate irradiation. Analysis of cell populations and cell surface molecules in whole body irradiated mice

    Ina, Yasuhiro; Sakai, Kazuo [Central Research Inst. of Electric Power Industry, Low Dose Radiation Research Center, Komae, Tokyo (Japan)

    2003-07-01

    The effects of low-dose rate whole body irradiation on biodefense and immunological systems were investigated using female C57BL/6 (B6) mice. These B6 mice were exposed continuously to {gamma}-rays from a {sup 137}Cs source in the long-term low-dose rate irradiation facility at CRIEPI for 0 - 12 weeks at a dose rate of 0.95 mGy/hr. In the bone marrow, thymus, spleen, lymph nodes, and peripheral blood of the irradiated mice, changes in cell populations and cell surface molecules were examined. The cell surface functional molecules (CD3, CD4, CD8, CD19, CD45R/B220, ICAM-1, Fas, NK-1.1, CXCR4, and CCR5), and activation molecules (THAM, CD28, CD40, CD44H, CD70, B7-1, B7-2, OX-40 antigen, CTLA-4, CD30 ligand, and CD40 ligand) were analyzed by flow cytometry. The percentage of CD4{sup +} T cells and cell surface CD8 molecule expressions on the CD8{sup +} T cells increased significantly to 120-130% after 3 weeks of the irradiation, compared to non-irradiated control mice. On the other hand, the percentage of CD45R/B220{sup +} CD40{sup +} B cells, which is one of the immunological markers of inflammation, infection, tumor, and autoimmune disease, decreased significantly to 80-90% between the 3rd to 5th week of irradiation. There was no significant difference in other cell population rates and cell surface molecule expression. Furthermore, abnormal T cells bearing mutated T cell receptors induced by high-dose rate irradiation were not observed throughout this study. These results suggest that low-dose rate irradiation activates the immunological status of the whole body. (author)

  13. Final Report - Montana State University - Microbial Activity and Precipitation at Solution-Solution Mixing Zones in Porous Media

    Gerlach, Robin [Montana State University

    2014-10-31

    Background. The use of biological and chemical processes that degrade or immobilize contaminants in subsurface environments is a cornerstone of remediation technology. The enhancement of biological and chemical processes in situ, involves the transport, displacement, distribution and mixing of one or more reactive agents. Biological and chemical reactions all require diffusive transport of solutes to reaction sites at the molecular scale and accordingly, the success of processes at the meter-scale and larger is dictated by the success of phenomena that occur at the micron-scale. However, current understanding of scaling effects on the mixing and delivery of nutrients in biogeochemically dynamic porous media systems is limited, despite the limitations this imposes on the efficiency and effectiveness of the remediation challenges at hand. Objectives. We therefore proposed to experimentally characterize and computationally describe the growth, evolution, and distribution of microbial activity and mineral formation as well as changes in transport processes in porous media that receive two or more reactive amendments. The model system chosen for this project was based on a method for immobilizing 90Sr, which involves stimulating microbial urea hydrolysis with ensuing mineral precipitation (CaCO3), and co-precipitation of Sr. Studies at different laboratory scales were used to visualize and quantitatively describe the spatial relationships between amendment transport and consumption that stimulate the production of biomass and mineral phases that subsequently modify the permeability and heterogeneity of porous media. Biomass growth, activity, and mass deposition in mixing zones was investigated using two-dimensional micro-model flow cells as well as flow cells that could be analyzed using synchrotron-based x-ray tomography. Larger-scale flow-cell experiments were conducted where the spatial distribution of media properties, flow, segregation of biological activity and

  14. Monitoring Cellular Interactions during T Cell Activation at the Single Molecule Level Using Semiconductor Quantum-Dots

    2005-05-10

    shift in the absorption and emission spectra for one of the tested shells, suggesting that exciton-molecular orbital (X-MO) coupling might take place...different bioconjugated qdots were compared. Testing the lipid raft hypothesis by single molecule imaging of targeted peptide-coated qdots: GPI- (lipid...The Rank Prize in opto-electronics The 2001 Michael and Kate Barany Biophysical Society Award Fellow of the Optical Society of America (1999).

  15. Shallow structure and its formation process of an active flexure in the forearc basin of the central Nankai subduction zone

    Ashi, J.; Ikehara, K.; Omura, A.; Ojima, T.; Murayama, M.

    2013-12-01

    ENE-WSW trending active faults, named Enshu fault system, are developed in the forearc basins of the eastern and central Nankai subduction zone. Three parallel faults developed in the Enshu forearc basin of the eastern Nankai have right lateral slip on the basis of dextral displacement of the canyon axis. Moreover, bathymetry data and side-scan sonar imageries indicate relative uplift of the northern region and the multichannel seismic (MCS) reflection profiles show northward dipping fault planes. In the central Nankai subuduction zone, an ENE-WSW trending step is distributed at the northern part of the Kumano forearc basin and is regarded as the western extension of the Enshu fault system. Although MCS records show deformations including an anticlinal fold beneath the bathymetric step, they have less resolution to identify deformation of basin sequence just below the seafloor. In contrast, deformation seems to reach to the seafloor on a profile by SBP mounted on a mother ship. Investigation of shallow deformation structures is significant for understanding of recent tectonic activity. We carried out deep towed SBP survey by ROV NSS (Navigable Sampling System) during Hakuho-maru KH-11-9 cruise. High resolution mapping of shallow structures was successfully conducted by a chirp SBP system of EdgeTech DW-106. ROV NSS also has capability to take a long core with a pinpoint accuracy around complex topographic region. The Kumano forearc basin is topographically divided into the northern part at a water depth of 2038 m and the other major region at a depth of 2042 m by the ENE-WSW linear step. Three deep towed SBP lines intersected this topographical step and revealed the following structures. This step is composed of 100 m wide gentle slope with an inclination of about 8 degrees. An anticlinal axis is located beneath the upper edge of this slope. Sedimentary layers continue at this slope region without any abut/termination and rapidly increase their thickness toward the

  16. Age, tectonic evolution and origin of the Aswa Shear Zone in Uganda: Activation of an oblique ramp during convergence in the East African Orogen

    Saalmann, K.; Mänttäri, I.; Nyakecho, C.; Isabirye, E.

    2016-05-01

    The Aswa Shear Zone (ASZ) is a major NW-SE trending structure of over 1000 km length in East Africa. In Uganda, the ASZ is a steeply NE-dipping, up to 11 km wide mylonitic shear zone that shows multiple stage brittle reactivation. On outcrop-scale, the fabric in the ASZ is characterized by a well-developed NW-SE striking and subvertical or steeply NE or SW dipping mylonitic foliation and a subhorizontal to moderately NW- or SE-plunging stretching lineation. Sinistral kinematics and fabric are very consistent along strike. The strain is heterogeneously distributed and partitioned into lens-shaped lower strain zones dominated by folding and characterized by pure shear, which are surrounded by high strain zones, some of them thick ultramylonites, with intense simple shear combined with flattening and strong transposition of pre-existing fabrics. Ductile shearing occurred during bulk E-W shortening, commenced at amphibolite facies conditions and continued with similar kinematics at greenschist and even lower grade conditions. A number of (sub-)parallel shear zones occur to the NE and SW of the main zone at a distance of up to 20-45 km. They show similar fabrics and kinematics and are thus related to activity along ASZ reflecting strain partitioning into simple shear and pure shear domains on a regional scale. Samples of mylonitic gneisses from the shear zone have been analyzed with U-Pb LA-MC-ICPMS and show Neoarchaean crystallisation ages between 2.66 and 2.61 Ga. Timing of ductile sinistral shearing is poorly constrained by lower intercept ages of 686 ± 62 and 640 ± 44 Ma. The fabric and structural relationship of the ca. 660 Ma Adjumani Granite exposed in the northern segment of ASZ suggest that the age of shear activity can be further limited to ca. 685 and 655 Ma. The Aswa Shear Zone is interpreted as an intra-cratonic, crustal-scale structure close to the northeastern margin of the Congo Craton, possibly inherited from previous continental extension. Early Aswa

  17. Neural progenitor cells isolated from the subventricular zone present hemichannel activity and form functional gap junctions with glial cells

    Talaverón, Rocío; Fernández, Paola; Escamilla, Rosalba; Pastor, Angel M.; Matarredona, Esperanza R.; Sáez, Juan C.

    2015-01-01

    The postnatal subventricular zone (SVZ) lining the walls of the lateral ventricles contains neural progenitor cells (NPCs) that generate new olfactory bulb interneurons. Communication via gap junctions between cells in the SVZ is involved in NPC proliferation and in neuroblast migration towards the olfactory bulb. SVZ NPCs can be expanded in vitro in the form of neurospheres that can be used for transplantation purposes after brain injury. We have previously reported that neurosphere-derived NPCs form heterocellular gap junctions with host glial cells when they are implanted after mechanical injury. To analyze functionality of NPC-glial cell gap junctions we performed dye coupling experiments in co-cultures of SVZ NPCs with astrocytes or microglia. Neurosphere-derived cells expressed mRNA for at least the hemichannel/gap junction channel proteins connexin 26 (Cx26), Cx43, Cx45 and pannexin 1 (Panx1). Dye coupling experiments revealed that gap junctional communication occurred among neurosphere cells (incidence of coupling: 100%). Moreover, hemichannel activity was also detected in neurosphere cells as evaluated in time-lapse measurements of ethidium bromide uptake. Heterocellular coupling between NPCs and glial cells was evidenced in co-cultures of neurospheres with astrocytes (incidence of coupling: 91.0 ± 4.7%) or with microglia (incidence of coupling: 71.9 ± 6.7%). Dye coupling in neurospheres and in co-cultures was inhibited by octanol, a gap junction blocker. Altogether, these results suggest the existence of functional hemichannels and gap junction channels in postnatal SVZ neurospheres. In addition, they demonstrate that SVZ-derived NPCs can establish functional gap junctions with astrocytes or microglia. Therefore, cell-cell communication via gap junctions and hemichannels with host glial cells might subserve a role in the functional integration of NPCs after implantation in the damaged brain. PMID:26528139

  18. Neural progenitor cells isolated from the subventricular zone present hemichannel activity and form functional gap junctions with glial cells.

    Talaverón, Rocío; Fernández, Paola; Escamilla, Rosalba; Pastor, Angel M; Matarredona, Esperanza R; Sáez, Juan C

    2015-01-01

    The postnatal subventricular zone (SVZ) lining the walls of the lateral ventricles contains neural progenitor cells (NPCs) that generate new olfactory bulb interneurons. Communication via gap junctions between cells in the SVZ is involved in NPC proliferation and in neuroblast migration towards the olfactory bulb. SVZ NPCs can be expanded in vitro in the form of neurospheres that can be used for transplantation purposes after brain injury. We have previously reported that neurosphere-derived NPCs form heterocellular gap junctions with host glial cells when they are implanted after mechanical injury. To analyze functionality of NPC-glial cell gap junctions we performed dye coupling experiments in co-cultures of SVZ NPCs with astrocytes or microglia. Neurosphere-derived cells expressed mRNA for at least the hemichannel/gap junction channel proteins connexin 26 (Cx26), Cx43, Cx45 and pannexin 1 (Panx1). Dye coupling experiments revealed that gap junctional communication occurred among neurosphere cells (incidence of coupling: 100%). Moreover, hemichannel activity was also detected in neurosphere cells as evaluated in time-lapse measurements of ethidium bromide uptake. Heterocellular coupling between NPCs and glial cells was evidenced in co-cultures of neurospheres with astrocytes (incidence of coupling: 91.0 ± 4.7%) or with microglia (incidence of coupling: 71.9 ± 6.7%). Dye coupling in neurospheres and in co-cultures was inhibited by octanol, a gap junction blocker. Altogether, these results suggest the existence of functional hemichannels and gap junction channels in postnatal SVZ neurospheres. In addition, they demonstrate that SVZ-derived NPCs can establish functional gap junctions with astrocytes or microglia. Therefore, cell-cell communication via gap junctions and hemichannels with host glial cells might subserve a role in the functional integration of NPCs after implantation in the damaged brain.

  19. Abnormal Activation of RhoA/ROCK-I Signaling in Junctional Zone Smooth Muscle Cells of Patients With Adenomyosis.

    Wang, S; Duan, H; Zhang, Y; Sun, F Q

    2016-03-01

    Adenomyosis (ADS) is a common estrogen-dependent gynecological disease with unknown etiology. The RhoA/Rho-kinase (ROCK) signaling pathway is involved in various cellular functions, including migration, proliferation, and smooth muscle contraction. Here we examined the potential role of this pathway in junctional zone (JZ) contraction in women with and without ADS. We demonstrated that in the normal JZ, RhoA and ROCK-I messenger RNA (mRNA) and protein expression was significantly higher in the proliferative phase of the menstrual cycle than in the secretory phase. Expression of RhoA and ROCK-I in the JZ from women with ADS was significantly higher than in the control women and showed no significant differences across the menstrual cycle. Treatment of JZ smooth muscle cells (JZSMCs) with estrogen at 0, 1, 10, or 100 nmol/L for 24 hours resulted in increased expression of RhoA, ROCK-I, and myosin light-chain (MLC) phosphorylation (p-MLC) in a dose-dependent manner. In parallel to its effects on p-MLC, estrogen-mediated, dose-dependent contraction responses in JZSMCs. Estrogen-mediated contraction in the ADS group was significantly higher than in the controls and also showed no significant differences across the menstrual cycle. These effects were suppressed in the presence of ICI 182780 or Y27632, supporting an estrogen receptor-dependent and RhoA activation-dependent mechanism. Our results indicate that the level of RhoA and ROCK-I increases in patients with ADS and the cyclic change is lost. Estrogen may affect uterine JZ contraction of ADS by enhancing RhoA/ ROCK-I signaling.

  20. Holocene activity and paleoseismicity of the Selaha Fault, southeastern segment of the strike-slip Xianshuihe Fault Zone, Tibetan Plateau

    Yan, Bing; Lin, Aiming

    2017-01-01

    In this study we examine the Holocene activity, including slip rate and paleoseismicity, of the Selaha Fault, a branch of the left-lateral strike-slip Xianshuihe Fault Zone located along the southeastern segment of the Ganzhi-Yushu-Xianshuihe Fault System (GYXFS) of the Tibetan Plateau. Interpretation of high-resolution images and field investigations reveal that the Selaha Fault is characterized by left-lateral strike-slip faulting with an average horizontal slip-rate of 9.0 mm/year during the Holocene. Trench excavations and 14C dating results show that at least three morphogenic earthquakes occurred during the past millennium; the most recent event occurred in the past 450 years and corresponds to the 1786 M 7.75 earthquake. The penultimate seismic event (E2) occurred in the period between 560 and 820 year BP (i.e., 1166-1428 CE) and is probably associated with the 1327 M 7.5 earthquake. The antepenultimate event (E3) is inferred to have occurred in the period between 820 ± 30 and 950 ± 30 year BP. Our results confirm that the Selaha Fault, as a portion of the GYXFS, plays an important role as a tectonic boundary in releasing the strain energy accumulated during the northeastward motion of the Tibetan Plateau in response to the ongoing northward penetration of the Indian Plate into the Eurasian Plate. The strain energy is released in the form of repeated large earthquakes that are recorded by strike-slip displacements of stream channels and alluvial fans.

  1. Single molecules and nanotechnology

    Vogel, Horst

    2007-01-01

    This book focuses on recent advances in the rapidly evolving field of single molecule research. These advances are of importance for the investigation of biopolymers and cellular biochemical reactions, and are essential to the development of quantitative biology. Written by leading experts in the field, the articles cover a broad range of topics, including: quantum photonics of organic dyes and inorganic nanoparticles their use in detecting properties of single molecules the monitoring of single molecule (enzymatic) reactions single protein (un)folding in nanometer-sized confined volumes the dynamics of molecular interactions in biological cells The book is written for advanced students and scientists who wish to survey the concepts, techniques and results of single molecule research and assess them for their own scientific activities.

  2. Upper plate deformation and seismic barrier in front of Nazca subduction zone: The Chololo Fault System and active tectonics along the Coastal Cordillera, southern Peru

    Audin, Laurence; Lacan, Pierre; Tavera, Hernando; Bondoux, Francis

    2008-11-01

    The South America plate boundary is one of the most active subduction zone. The recent Mw = 8.4 Arequipa 2001 earthquake ruptured the subduction plane toward the south over 400 km and stopped abruptly on the Ilo Peninsula. In this exact region, the subduction seismic crisis induced the reactivation of continental fault systems in the coastal area. We studied the main reactivated fault system that trends perpendicular to the trench by detailed mapping of fault related-geomorphic features. Also, at a longer time scale, a recurrent Quaternary transtensive tectonic activity of the CFS is expressed by offset river gullies and alluvial fans. The presence of such extensional fault systems trending orthogonal to the trench along the Coastal Cordillera in southern Peru is interpreted to reflect a strong coupling between the two plates. In this particular case, stress transfer to the upper plate, at least along the coastal fringe, appears to have induced crustal seismic events that were initiated mainly during and after the 2001 earthquake. The seafloor roughness of the subducting plate is usually thought to be a cause of segmentation along subduction zones. However, after comparing and discussing the role of inherited structures within the upper plate to the subduction zone segmentation in southern Peru, we suggest that the continental structure itself may exert some feedback control on the segmentation of the subduction zone and thus participate to define the rupture pattern of major subduction earthquakes along the southern Peru continental margin.

  3. Fluid flow and water-rock interaction across the active Nankai Trough subduction zone forearc revealed by boron isotope geochemistry

    Hüpers, Andre; Kasemann, Simone A.; Kopf, Achim J.; Meixner, Anette; Toki, Tomohiro; Shinjo, Ryuichi; Wheat, C. Geoffrey; You, Chen-Feng

    2016-11-01

    Compositional changes, dehydration reactions and fluid flow in subducted sediments influence seismogenesis and arc magmatism in subduction zones. To identify fluid flow and water-rock interaction processes in the western Nankai Trough subduction zone (SW Japan) we analyzed boron concentration and boron isotope composition (δ11B) of pore fluids sampled across the subduction zone forearc from depths of up to ∼922 m below seafloor during four Integrated Ocean Drilling Program (IODP) Expeditions. The major structural regimes that were sampled by coring include: (1) sedimentary inputs, (2) the frontal thrust zone, (3) the megasplay fault zone, and (4) the forearc basin. From mass balance consideration we find that consumption of boron (B) by ash alteration and desorption of B from the solid phase, mediated by organic matter degradation, produces a net decrease in B concentrations with depth down to ∼120 μM and variable δ11B values in the range of ∼+20‰ and +49‰. Interstitial water in sediments on the incoming oceanic plate are influenced by more efficient mobilization of exchangeable B from the solid phase due to higher temperatures and alteration of the oceanic crust that acts as a sink for 10B. At the tip of the megasplay fault zone, elevated B concentration and B isotopic composition suggest that underthrust coarse-grained slope sediments provide a pathway for fluids out of the upper (balance considerations suggest a shallower fluid source depth compared to pore fluids sampled previously near the décollement zone along the central portion of the Nankai margin.

  4. Organic molecules as tools to control the growth, surface structure, and redox activity of colloidal quantum dots.

    Weiss, Emily A

    2013-11-19

    In order to achieve efficient and reliable technology that can harness solar energy, the behavior of electrons and energy at interfaces between different types or phases of materials must be understood. Conversion of light to chemical or electrical potential in condensed phase systems requires gradients in free energy that allow the movement of energy or charge carriers and facilitate redox reactions and dissociation of photoexcited states (excitons) into free charge carriers. Such free energy gradients are present at interfaces between solid and liquid phases or between inorganic and organic materials. Nanostructured materials have a higher density of these interfaces than bulk materials. Nanostructured materials, however, have a structural and chemical complexity that does not exist in bulk materials, which presents a difficult challenge: to lower or eliminate energy barriers to electron and energy flux that inevitably result from forcing different materials to meet in a spatial region of atomic dimensions. Chemical functionalization of nanostructured materials is perhaps the most versatile and powerful strategy for controlling the potential energy landscape of their interfaces and for minimizing losses in energy conversion efficiency due to interfacial structural and electronic defects. Colloidal quantum dots are semiconductor nanocrystals synthesized with wet-chemical methods and coated in organic molecules. Chemists can use these model systems to study the effects of chemical functionalization of nanoscale organic/inorganic interfaces on the optical and electronic properties of a nanostructured material, and the behavior of electrons and energy at interfaces. The optical and electronic properties of colloidal quantum dots have an intense sensitivity to their surface chemistry, and their organic adlayers make them dispersible in solvent. This allows researchers to use high signal-to-noise solution-phase spectroscopy to study processes at interfaces. In this

  5. Cardiotrophin-1 induces intercellular adhesion molecule-1 expression by nuclear factor κB activation in human umbilical vein endothelial cells

    2008-01-01

    Background In addition to elevated concentrations of cytokines, patients with congestive heart failure (CHF) show endothelial dysfunction and increased plasma concentrations of adhesion molecules like intercellular adhesion molecule-1 (ICAM-1). Furthermore, the concentration of cardiotrophin-1 (CT-1) - a cytokine of the interleukin-6 superfamily - is increased in CHF. We tested the hypothesis whether CT-1 is able to induce ICAM-1 in human umbilical vein endothelial cells (HUVEC). Furthermore we examined the signalling mechanisms of CT-1 mediated ICAM-1 expression. Methods Confluent layers of HUVEC were incubated with increasing concentrations of CT-1 (5 to 100 ng/ml) for different periods. ICAM-1 mRNA was determined by real-time polymerase chain reaction (PCR) and ICAM-1 surface expression by fluorescence-activated cell sorter (FACS) analysis and soluble ICAM-1 (slCAM-1) in the culture supematant by enzyme linked immunosorbent assay (ELISA). To clarify the signalling pathway of CT-1 induced ICAM-1 expression we used various inhibitors of possible signal transducing molecules, electromobility shift assay (EMSA) and Western blot analysis. Results CT-1 induced ICAM-1 mRNA (1.8i-0.8 fold increase compared to unstimulated cells after 6 hours) and protein (1.4~-0.2 fold increase compared to unstimulated cells after 48 hours) in HUVEC in a time- and concentration-dependent manner. EMSA experiments show that CT-1 causes nuclear factor (NF) KB activation. Because parthenolide could inhibit CT-1 induced ICAM-1 expression NFKB activation is required in this pathway. CT-1 did not activate extraceUular signal regulated kinases (ERK), c-Jun N-terminal kinase (JNK) and p38. Conclusion CT-1 is able to induce ICAM-1 in endothelial cells by NFKB activation. These results may explain in part elevated ICAM-1 concentrations in patients with CHF and endothelial dysfunction.

  6. Structure–activity relationships of a small-molecule inhibitor of the PDZ domain of PICK1

    Bach, Anders; Stuhr-Hansen, Nicolai; Thorsen, Thor S.

    2010-01-01

    Recently, we described the first small-molecule inhibitor, (E)-ethyl 2-cyano-3-(3,4-dichlorophenyl)acryloylcarbamate (1), of the PDZ domain of protein interacting with Ca-kinase 1 (PICK1), a potential drug target against brain ischemia, pain and cocaine addiction. Herein, we explore structure......, docking studies were used to correlate biological affinities with structural considerations for ligand–protein interactions. The most potent analogue obtained in this study showed an improvement in affinity compared to 1 and is currently a lead in further studies of PICK1 inhibition....

  7. Coordination geometries of Zn(II) and Cd(II) in phosphotriesterase: Influence of water molecules in the active site

    Krauss, M; Olsen, Lars; Antony, J;

    2002-01-01

    to the Od1 of the carboxylate of the first-shell aspartate designated M 1, but the energy difference between Cd1Zn2 and the lowest energy Zn1Cd2 structure is only about 2 kcal/mol and decreasing with the addition of water molecules. The Zn1Cd2 arrangement is found experimentally....... performed to analyze the experimentally determined chemical shifts at 212 and 116 ppm, respectively, for the CdCd enzyme. The calculated isotropic shieldings correlate with the coordination number of the metal ions, indicating that the CdCd enzyme has one more ligand at the high shift site than at the low...

  8. Presence and patterns of alkaline phosphatase activity and phosphorus cycling in natural riparian zones under changing nutrient conditions

    Peifang Wang

    2014-08-01

    Full Text Available Phosphorus (P is an important limiting nutrient in aquatic ecosystems and knowledge of P cycling is fundamental for reducing harmful algae blooms and other negative effects in water. Despite their importance, the characteristics of P cycling under changing nutrient conditions in shallow lakes were poorly investigated. In this study, in situ incubation experiments were conducted in a natural riparian zone in the main diversion channel used for water transfer into Lake Taihu (Wangyu River. Variations in microbial biomass, dissolved P fractions (organic and inorganic, and alkaline phosphatase activity (bulk APA and specific APA were determined after incubation with and without the addition of P and nitrogen (N (4 total water treatments: +P, +N, +NP, and control. Experiments were conducted during two seasons (late spring and early fall to account for natural differences in nutrient levels that may occur in situ. Our results demonstrated that low levels of DRP may not necessarily indicate P limitation. Phytoplankton exhibited “serial N limitation with P stress” in May, such that chlorophyll a (Chl a increased significantly with N addition, while the limiting nutrient shifted to P in October and phytoplankton biomass increased with P addition. Phytoplankton contributed greatly to APA production and was significantly influenced by P bioavailability, yet high levels of bulk APA were also not necessarily indicative of P limitation. In contrast to phytoplankton, bacteria were less P stressed. As a consequence of enhanced utilization of dissolved reactive P (DRP and dissolved organic P (DOP, +N treatment elevated APA significantly. By contrast, APA could be repressed to low values and phytoplankton converted a large portion of DRP to DOP with P addition. But this was not consistent with bacteria APA (bact-APA in the absence or presence of abundant phytoplankton biomass. The correlation between bulk APA and DRP was good at separate sites and discrepant

  9. Activated regulatory T cell regulates neural stem cell proliferation in the subventricular zone of normal and ischemic mouse brain through interleukin 10

    Jixian eWang

    2015-09-01

    Full Text Available Recent studies have demonstrated that the depletion of Regulatory T cells (Tregs inhibits neural progenitor cell migration after brain ischemia. However, whether Tregs affect neural stem/progenitor cell proliferation is unclear. We explored the effect of Tregs on neurogenesis in the subventricular zone after ischemia. Tregs were isolated and activated in vitro. Adult male C57BL/6 mice underwent 60 minutes transient middle cerebral artery occlusion (tMCAO. Then Tregs (1x105 were injected into the left lateral ventricle of normal and ischemic mouse brain. Neurogenesis was determined by immunostaining. The mechanism was examined by inhibiting interleukin 10 (IL-10 and transforming growth factor (TGF- signaling. We found that the number of BrdU+ cells in the subventricular zone was significantly increased in the activated Tregs-treated mice. Double immunostaining showed that these BrdU+ cells expressed Mash1. Blocking IL-10 reduced the number of Mash1+/BrdU+ cells, but increased the amount of GFAP+/BrdU+ cells. Here we conclude that activated Tregs enhanced neural stem cell proliferation in the subventricular zone of normal and ischemic mice; blockage of IL-10 abolished Tregs-mediated neural stem cell proliferation in vivo and in vitro. Our results suggest that activated Tregs promoted neural stem cell proliferation via IL-10, which provides a new therapeutic approach for ischemic stroke.

  10. Synthesis of Zn-MOF incorporating titanium-hydrides as active sites binding H{sub 2} molecules

    Kim, Jongsik, E-mail: jkim40@nd.edu [Department of Chemical and Biomolecular Engineering, University of Notre Dame, 182, Fitzpatrick Hall, Notre Dame, IN 46556 (United States); Ok Kim, Dong; Wook Kim, Dong; Sagong, Kil [Hanwha Chemical Research & Development Center, 6, Shinseong-dong, Yuseong-gu, Daejeon 305-804 (Korea, Republic of)

    2015-10-15

    This paper describes the synthetic effort for a Zn-MOF imparting Ti-H as a preferential binding site potentially capturing H{sub 2} molecules via Kubas-type interaction. The formation mechanism of Ti-H innate to the final material was potentially demonstrated to follow a radical dissociation rather than a β-hydrogen elimination and a C-H reductive elimination. - Graphical abstract: This study details the synthesis and the formation mechanism of Zn-MOF adsorbent site-isolating TiH{sub 3} that can potentially capture H{sub 2} molecules via Kubas-binding mechanism. - Highlights: • OH-functionalized Zn-MOF was employed as a reactive template to site-isolate TiH{sub 3}. • This MOF was post-synthetically modified using a tetracyclohexyl titanium (IV). • This intermediate was hydrogenolyzed to change ligand from cyclohexyl to hydride. • Formation mechanism of TiH{sub 3} was investigated via two control GC–MS experiments. • Final Zn-MOF potentially site-isolating TiH{sub 3} species was used as a H{sub 2} adsorbent.

  11. The 5'-flanking region of the RP58 coding sequence shows prominent promoter activity in multipolar cells in the subventricular zone during corticogenesis.

    Ohtaka-Maruyama, C; Hirai, S; Miwa, A; Takahashi, A; Okado, H

    2012-01-10

    Pyramidal neurons of the neocortex are produced from progenitor cells located in the neocortical ventricular zone (VZ) and subventricular zone (SVZ) during embryogenesis. RP58 is a transcriptional repressor that is strongly expressed in the developing brain and plays an essential role in corticogenesis. The expression of RP58 is strictly regulated in a time-dependent and spatially restricted manner. It is maximally expressed in E15-16 embryonic cerebral cortex, localized specifically to the cortical plate and SVZ of the neocortex, hippocampus, and parts of amygdala during brain development, and found in glutamatergic but not GABAergic neurons. Identification of the promoter activity underlying specific expression patterns provides important clues to their mechanisms of action. Here, we show that the RP58 gene promoter is activated prominently in multipolar migrating cells, the first in vivo analysis of RP58 promoter activity in the brain. The 5.3 kb 5'-flanking genomic DNA of the RP58 coding region demonstrates promoter activity in neurons both in vitro and in vivo. This promoter is highly responsive to the transcription factor neurogenin2 (Ngn2), which is a direct upstream activator of RP58 expression. Using in utero electroporation, we demonstrate that RP58 gene promoter activity is first detected in a subpopulation of pin-like VZ cells, then prominently activated in migrating multipolar cells in the multipolar cell accumulation zone (MAZ) located just above the VZ. In dissociated primary cultured cortical neurons, RP58 promoter activity mimics in vivo expression patterns from a molecular standpoint that RP58 is expressed in a fraction of Sox2-positive progenitor cells, Ngn2-positive neuronal committed cells, and Tuj1-positive young neurons, but not in Dlx2-positive GABAergic neurons. Finally, we show that Cre recombinase expression under the control of the RP58 gene promoter is a feasible tool for conditional gene switching in post-mitotic multipolar migrating

  12. Cathelicidin-like helminth defence molecules (HDMs: absence of cytotoxic, anti-microbial and anti-protozoan activities imply a specific adaptation to immune modulation.

    Karine Thivierge

    Full Text Available Host defence peptides (HDPs are expressed throughout the animal and plant kingdoms. They have multifunctional roles in the defence against infectious agents of mammals, possessing both bactericidal and immune-modulatory activities. We have identified a novel family of molecules secreted by helminth parasites (helminth defence molecules; HDMs that exhibit similar structural and biochemical characteristics to the HDPs. Here, we have analyzed the functional activities of four HDMs derived from Schistosoma mansoni and Fasciola hepatica and compared them to human, mouse, bovine and sheep HDPs. Unlike the mammalian HDPs the helminth-derived HDMs show no antimicrobial activity and are non-cytotoxic to mammalian cells (macrophages and red blood cells. However, both the mammalian- and helminth-derived peptides suppress the activation of macrophages by microbial stimuli and alter the response of B cells to cytokine stimulation. Therefore, we hypothesise that HDMs represent a novel family of HDPs that evolved to regulate the immune responses of their mammalian hosts by retaining potent immune modulatory properties without causing deleterious cytotoxic effects.

  13. Upper plate deformation and seismic barrier in front of Nazca subduction zone : the Chololo Fault System and active tectonics along the Coastal Cordillera, southern Peru

    Audin, Laurence; Lacan, P.; Tavera, H.; Bondoux, Francis

    2008-01-01

    The South America plate boundary is one of the most active subduction zone. The recent Mw=8.4 Arequipa 2001 earthquake ruptured the subduction plane toward the south over 400 km and stopped abruptly on the Ilo Peninsula. In this exact region, the subduction seismic crisis induced the reactivation of continental fault systems in the coastal area. We studied the main reactivated fault system that trends perpendicular to the trench by detailed mapping of fault related-geomorphic features. Also, ...

  14. Glucuronidation of odorant molecules in the rat olfactory system: activity, expression and age-linked modifications of UDP-glucuronosyltransferase isoforms, UGT1A6 and UGT2A1, and relation to mitral cell activity.

    Leclerc, Séverine; Heydel, Jean-Marie; Amossé, Valérie; Gradinaru, Daniela; Cattarelli, Martine; Artur, Yves; Goudonnet, Hervé; Magdalou, Jacques; Netter, Patrick; Pelczar, Hélène; Minn, Alain

    2002-11-15

    The aim of the present study was to examine the glucuronidation of a series of odorant molecules by homogenates prepared either with rat olfactory mucosa, olfactory bulb or brain. Most of the odorant molecules tested were efficiently conjugated by olfactory mucosa, whereas olfactory bulb and brain homogenates displayed lower activities and glucuronidated only a few molecules. Important age-related changes in glucuronidation efficiency were observed in olfactory mucosa and bulb. Therefore, we studied changes in expression of two UDP-glucuronosyltransferase isoforms, UGT1A6 and UGT2A1, in 1-day, 1- and 2-week-, 3-, 12- and 24-month-old rats. UGT1A6 was expressed at the same transcriptional level in the olfactory mucosa, bulb and brain, throughout the life period studied. UGT2A1 mRNA was expressed in both olfactory mucosa and olfactory bulb, in accordance with previous results [Mol. Brain Res. 90 (2001) 83], but UGT2A1 transcriptional level was 400-4000 times higher than that of UGT1A6. Moreover, age-dependent variations in UGT2A1 mRNA expression were observed. As it has been suggested that drug metabolizing enzymes could participate in olfactory function, mitral cell electrical activity was recorded during exposure to different odorant molecules in young, adult and old animals. Age-related changes in the amplitude of response after stimulation with several odorant molecules were observed, and the highest responses were obtained with molecules that were not efficiently glucuronidated by olfactory mucosa. In conclusion, the present work presents new evidence of the involvement of UGT activity in some steps of the olfactory process.

  15. [Autoradiographic investigations on postnatal proliferative activity of the telencephalic and diencephalic matrix-zones in the axolotl (Ambystoma mexicanum), with special references to the olfactory organ (author's transl)].

    Richter, W; Kranz, D

    1981-01-01

    The localization and proliferative activity of the matrix-zones has been investigated in the telencephalon and in the diencephalon of 21 axolotls (Ambystoma mexicanum) by means of autoradiographs, after injection of tritiated thymidine at different stages of the postnatal life. There are no previous detailed autoradiographical reports on postnatal brain development in the axolotl. Matrix-zones (i.e. ventricular and subventricular zone) exist in the dorsal part and in the ventral part of the telencephalon, we have found these also in the diencephalon in the wall of the preoptic recessus and ventrally of the habenula. The quantitative part of this study indicates high values of the labeling-index in the early postnatal stages. Then, the labeling-index decreases, but also in 3 years old specimens labeled cells were observed in the matrix-zones of the telencephalon; therefore a few of proliferative capacity remains in the central nervous system of adult axolotls. Labeled cells were also found in the olfactory organ of early postnatal and adult axolotls; these are neuroblasts which have relevance for the regeneration of the forebrain.

  16. De-repression of the RAC activator ELMO1 in cancer stem cells drives progression of TGFβ-deficient squamous cell carcinoma from transition zones

    McCauley, Heather A; Chevrier, Véronique; Birnbaum, Daniel; Guasch, Géraldine

    2017-01-01

    Squamous cell carcinomas occurring at transition zones are highly malignant tumors with poor prognosis. The identity of the cell population and the signaling pathways involved in the progression of transition zone squamous cell carcinoma are poorly understood, hence representing limited options for targeted therapies. Here, we identify a highly tumorigenic cancer stem cell population in a mouse model of transitional epithelial carcinoma and uncover a novel mechanism by which loss of TGFβ receptor II (Tgfbr2) mediates invasion and metastasis through de-repression of ELMO1, a RAC-activating guanine exchange factor, specifically in cancer stem cells of transition zone tumors. We identify ELMO1 as a novel target of TGFβ signaling and show that restoration of Tgfbr2 results in a complete block of ELMO1 in vivo. Knocking down Elmo1 impairs metastasis of carcinoma cells to the lung, thereby providing insights into the mechanisms of progression of Tgfbr2-deficient invasive transition zone squamous cell carcinoma. DOI: http://dx.doi.org/10.7554/eLife.22914.001 PMID:28219480

  17. Radiant zone heated particulate filter

    Gonze, Eugene V [Pinckney, MI

    2011-12-27

    A system includes a particulate matter (PM) filter including an upstream end for receiving exhaust gas and a downstream end. A radiant zoned heater includes N zones, where N is an integer greater than one, wherein each of the N zones includes M sub-zones, where M is an integer greater than or equal to one. A control module selectively activates at least a selected one of the N zones to initiate regeneration in downstream portions of the PM filter from the one of the N zones, restricts exhaust gas flow in a portion of the PM filter that corresponds to the selected one of the N zones, and deactivates non-selected ones of the N zones.

  18. Benzimidazole derivative small-molecule 991 enhances AMPK activity and glucose uptake induced by AICAR or contraction in skeletal muscle

    Bultot, Laurent; Jensen, Thomas Elbenhardt; Lai, Yu-Chiang

    2016-01-01

    be beneficial for both purposes. Here, we investigated if a recently described potent AMPK activator called 991, in combination with the commonly used activator 5-aminoimidazole-4-carboxamide riboside or contraction, further enhances AMPK activity and glucose transport in mouse skeletal muscle ex vivo. Given...... that the γ3-subunit is exclusively expressed in skeletal muscle and has been implicated in contraction-induced glucose transport, we measured the activity of AMPKγ3 as well as ubiquitously expressed γ1-containing complexes. We initially validated the specificity of the antibodies for the assessment...... profoundly enhanced AMPKγ1/γ3 complex activation and glucose transport compared with any of the single treatments. The study demonstrates the utility of a dual activator approach to achieve a greater activation of AMPK and downstream physiological responses in various cell types, including skeletal muscle....

  19. Catalytic enantioselective construction of quaternary stereocenters: assembly of key building blocks for the synthesis of biologically active molecules.

    Liu, Yiyang; Han, Seo-Jung; Liu, Wen-Bo; Stoltz, Brian M

    2015-03-17

    The ever-present demand for drugs with better efficacy and fewer side effects continually motivates scientists to explore the vast chemical space. Traditionally, medicinal chemists have focused much attention on achiral or so-called "flat" molecules. More recently, attention has shifted toward molecules with stereogenic centers since their three-dimensional structures represent a much larger fraction of the chemical space and have a number of superior properties compared with flat aromatic compounds. Quaternary stereocenters, in particular, add greatly to the three-dimensionality and novelty of the molecule. Nevertheless, synthetic challenges in building quaternary stereocenters have largely prevented their implementation in drug discovery. The lack of effective and broadly general methods for enantioselective formation of quaternary stereocenters in simple molecular scaffolds has prompted us to investigate new chemistry and develop innovative tools and solutions. In this Account, we describe three approaches to constructing quaternary stereocenters: nucleophilic substitution of 3-halooxindoles, conjugate addition of boronic acids to cyclic enones, and allylic alkylation of enolates. In the first approach, malonic ester nucleophiles attack electrophilic 3-halooxindoles, mediated by a copper(II)-bisoxazoline catalyst. A variety of oxindoles containing a benzylic quaternary stereocenter can be accessed through this method. However, it is only applicable to the specialized 3,3-disubstituted oxindole system. To access benzylic quaternary stereocenters in a more general context, we turned our attention to the enantioselective conjugate addition of carbon nucleophiles to α,β-unsaturated carbonyl acceptors. We discovered that in the presence of catalytic palladium-pyridinooxazoline complex, arylboronic acids add smoothly to β-substituted cyclic enones to furnish ketones with a β-benzylic quaternary stereocenter in high yields and enantioselectivities. The reaction is

  20. Soluble Collectin-12 (CL-12) Is a Pattern Recognition Molecule Initiating Complement Activation via the Alternative Pathway

    Ma, Ying Jie; Hein, Estrid; Munthe-Fog, Lea

    2015-01-01

    and may recognize certain bacteria and fungi, leading to opsonophagocytosis. However, based on its structural and functional similarities with soluble collectins, we hypothesized the existence of a fluid-phase analog of CL-12 released from cells, which may function as a soluble pattern-recognition...... of the terminal complement complex. These results demonstrate the existence of CL-12 in a soluble form and indicate a novel mechanism by which the alternative pathway of complement may be triggered directly by a soluble pattern-recognition molecule....... nonreducing conditions it presented multimeric assembly forms. Immunoprecipitation and Western blot analysis of human umbilical cord plasma enabled identification of a natural soluble form of CL-12 having an electrophoretic mobility pattern close to that of shed soluble recombinant CL-12. Soluble CL-12 could...

  1. Identification of a novel Arabidopsis thaliana nitric oxide-binding molecule with guanylate cyclase activity in vitro

    Mulaudzi, Takalani

    2011-09-01

    While there is evidence of nitric oxide (NO)-dependent signalling via the second messenger cyclic guanosine 3′,5′-monophosphate (cGMP) in plants, guanylate cyclases (GCs), enzymes that catalyse the formation of cGMP from guanosine 5′-triphosphate (GTP) have until recently remained elusive and none of the candidates identified to-date are NO-dependent. Using both a GC and heme-binding domain specific (H-NOX) search motif, we have identified an Arabidopsis flavin monooxygenase (At1g62580) and shown electrochemically that it binds NO, has a higher affinity for NO than for O 2 and that this molecule can generate cGMP from GTP in vitro in an NO-dependent manner. © 2011 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

  2. Optimized geometry, vibration (IR and Raman spectra and nonlinear optical activity of p-nitroanilinium perchlorate molecule: A theoretical study

    Tamer Ömer

    2016-03-01

    Full Text Available The molecular modeling of p-nitroanilinium perchlorate molecule was carried out by using B3LYP and HSEH1PBE levels of density functional theory (DFT. The IR and Raman spectra were simulated and the assignments of vibrational modes were performed on the basis of relative contribution of various internal co-ordinates. NBO analysis was performed to demonstrate charge transfer, conjugative interactions and the formation of intramolecular hydrogen bonding interactions within PNAPC. Obtained large dipole moment values showed that PNAPC is a highly polarizable complex, and the charge transfer occurs within PNAPC. Hydrogen bonding and charge transfer interactions were also displayed by small HOMO-LUMO gap and molecular electrostatic potential (MEP surface. The strong evidences that the material can be used as an efficient nonlinear optical (NLO material of PNAPC were demonstrated by considerable polarizability and hyperpolarizability values obtained at DFT levels.

  3. New design strategy for the two-photon active material based on push-pull substituted bisanthene molecule.

    Chattopadhyaya, Mausumi; Alam, Md Mehboob; Chakrabarti, Swapan

    2011-03-31

    In the present work, we have critically examined the origin of strong two-photon transition probability of a donor-acceptor substituted bisanthene molecule that imitates a small piece of edge passivated (4, 4) graphene nanoribbon. In our calculations, we have considered -OMe, and -NH(2) as donors and -NO(2) as an acceptor. The one- and two-photon absorption parameters are evaluated using state-of-the-art linear and quadratic response theory, respectively, and all these calculations are carried out within the framework of time dependent density functional theory. To give a proper judgment on our findings, we have used the long-range corrected CAMB3LYP functional for all of the time dependent calculations. The present investigation reveals that the bisanthene molecule with three pairs of donor/acceptor moiety has a lower two-photon transition probability than that of a suitably designed bisanthene with only a single pair of donor/acceptor moiety. This in silico observation is consistent for all of the donor/acceptor moieties chosen in the present work. A comprehensive analysis at the two state model level of theory clearly offers us a verdict that by placing the donor/acceptor moiety in a suitable position of bisanthene, we can create a significant asymmetry in the electron density in the first excited state, which eventually leads to a significant difference in the ground and excited state dipole moment and is attributed to the higher two-photon transition probability of a particular bisanthene with a single pair of donor/acceptor moiety than bisanthene with three pairs of donor/acceptor.

  4. The adaptor molecule signaling lymphocytic activation molecule (SLAM)-associated protein (SAP) is essential in mechanisms involving the Fyn tyrosine kinase for induction and progression of collagen-induced arthritis.

    Zhong, Ming-Chao; Veillette, André

    2013-11-01

    Signaling lymphocytic activation molecule-associated protein (SAP) is an Src homology 2 domain-only adaptor involved in multiple immune cell functions. It has also been linked to immunodeficiencies and autoimmune diseases, such as systemic lupus erythematosus. Here, we examined the role and mechanism of action of SAP in autoimmunity using a mouse model of autoimmune arthritis, collagen-induced arthritis (CIA). We found that SAP was essential for development of CIA in response to collagen immunization. It was also required for production of collagen-specific antibodies, which play a key role in disease pathogenesis. These effects required SAP expression in T cells, not in B cells. In mice immunized with a high dose of collagen, the activity of SAP was nearly independent of its ability to bind the protein tyrosine kinase Fyn and correlated with the capacity of SAP to promote full differentiation of follicular T helper (TFH) cells. However, with a lower dose of collagen, the role of SAP was more dependent on Fyn binding, suggesting that additional mechanisms other than TFH cell differentiation were involved. Further studies suggested that this might be due to a role of the SAP-Fyn interaction in natural killer T cell development through the ability of SAP-Fyn to promote Vav-1 activation. We also found that removal of SAP expression during progression of CIA attenuated disease severity. However, it had no effect on disease when CIA was clinically established. Together, these results indicate that SAP plays an essential role in CIA because of Fyn-independent and Fyn-dependent effects on TFH cells and, possibly, other T cell types.

  5. The Mediation of Learning in the Zone of Proximal Development through a Co-Constructed Writing Activity

    Thompson, Ian

    2013-01-01

    This article develops a theoretical understanding of the processes involved in the co-construction of a written text by a teacher and student from a Vygotskian perspective. Drawing on cultural-historical and sociocultural theories of writing and Vygotsky's concept of the Zone of Proximal Development (ZPD), this case study of a student and teacher…

  6. 77 FR 36997 - Foreign-Trade Zone 61-San Juan, PR; Notification of Proposed Production Activity; Pfizer...

    2012-06-20

    ... pharmaceutical products in bulk mixture or dosage form under FTZ procedures at the Guayama site. Production under... (Subzone 61A); (Ibuprofen Pharmaceutical Products); Guayama, PR The Puerto Rico Trade and Export Company... various pharmaceutical products under zone procedures (Board Order 617, 12/11/ 1992, 57 FR 61046,...

  7. SLITHER: a web server for generating contiguous conformations of substrate molecules entering into deep active sites of proteins or migrating through channels in membrane transporters.

    Lee, Po-Hsien; Kuo, Kuei-Ling; Chu, Pei-Ying; Liu, Eric M; Lin, Jung-Hsin

    2009-07-01

    Many proteins use a long channel to guide the substrate or ligand molecules into the well-defined active sites for catalytic reactions or for switching molecular states. In addition, substrates of membrane transporters can migrate to another side of cellular compartment by means of certain selective mechanisms. SLITHER (http://bioinfo.mc.ntu.edu.tw/slither/or http://slither.rcas.sinica.edu.tw/) is a web server that can generate contiguous conformations of a molecule along a curved tunnel inside a protein, and the binding free energy profile along the predicted channel pathway. SLITHER adopts an iterative docking scheme, which combines with a puddle-skimming procedure, i.e. repeatedly elevating the potential energies of the identified global minima, thereby determines the contiguous binding modes of substrates inside the protein. In contrast to some programs that are widely used to determine the geometric dimensions in the ion channels, SLITHER can be applied to predict whether a substrate molecule can crawl through an inner channel or a half-channel of proteins across surmountable energy barriers. Besides, SLITHER also provides the list of the pore-facing residues, which can be directly compared with many genetic diseases. Finally, the adjacent binding poses determined by SLITHER can also be used for fragment-based drug design.

  8. The fibroblast growth factor receptor (FGFR) agonist FGF1 and the neural cell adhesion molecule-derived peptide FGL activate FGFR substrate 2alpha differently

    Chen, Yongshuo; Li, Shizhong; Berezin, Vladimir

    2010-01-01

    Activation of fibroblast growth factor (FGF) receptors (FGFRs) both by FGFs and by the neural cell adhesion molecule (NCAM) is crucial in the development and function of the nervous system. We found that FGFR substrate 2alpha (FRS2alpha), Src homologous and collagen A (ShcA), and phospholipase......-Cgamma (PLCgamma) were all required for neurite outgrowth from cerebellar granule neurons (CGNs) induced by FGF1 and FGL (an NCAM-derived peptide agonist of FGFR1). Like FGF1, FGL induced tyrosine phosphorylation of FGFR1, FRS2alpha, ShcA, and PLCgamma in a time- and dose-dependent manner. However, the activation...... of FRS2alpha by FGL was significantly lower than the activation by FGF1, indicating a differential signaling profile induced by NCAM compared with the cognate growth factor....

  9. Role of an expansin-like molecule in Dictyostelium morphogenesis and regulation of its gene expression by the signal transducer and activator of transcription protein Dd-STATa.

    Ogasawara, Shun; Shimada, Nao; Kawata, Takefumi

    2009-02-01

    Expansins are proteins involved in plant morphogenesis, exerting their effects on cellulose to extend cell walls. Dictyostelium is an organism that possesses expansin-like molecules, but their functions are not known. In this study, we analyzed the expL7 (expansin-like 7) gene, which has been identified as a putative target of Dd-STATa, a Dictyostelium homolog of the metazoan signal transducer and activator of transcription (STAT) proteins. Promoter fragments of the expL7 were fused to a lacZ reporter and the expression patterns determined. As expected from the behavior of the endogenous expL7 gene, the expL7/lacZ fusion gene was downregulated in Dd-STATa null slugs. In the parental strain, the expL7 promoter was activated in the anterior tip region. Mutational analysis of the promoter identified a sequence that was necessary for expression in tip cells. In addition, an activator sequence for pstAB cells was identified. These sequences act in combination with the repressor region to prevent ectopic expL7 expression in the prespore and prestalk regions of the slug and culminant. Although the expL7 null mutant showed no phenotypic change, the expL7 overexpressor showed aberrant stalk formation. These results indicate that the expansin-like molecule is important for morphogenesis in Dictyostelium.

  10. Nanodevices for generating power from molecules and batteryless sensing

    Wang, Yinmin; Wang, Xianying; Hamza, Alex V.

    2017-01-03

    A nanoconverter or nanosensor is disclosed capable of directly generating electricity through physisorption interactions with molecules that are dipole containing organic species in a molecule interaction zone. High surface-to-volume ratio semiconductor nanowires or nanotubes (such as ZnO, silicon, carbon, etc.) are grown either aligned or randomly-aligned on a substrate. Epoxy or other nonconductive polymers are used to seal portions of the nanowires or nanotubes to create molecule noninteraction zones. By correlating certain molecule species to voltages generated, a nanosensor may quickly identify which species is detected. Nanoconverters in a series parallel arrangement may be constructed in planar, stacked, or rolled arrays to supply power to nano- and micro-devices without use of external batteries. In some cases breath, from human or other life forms, contain sufficient molecules to power a nanoconverter. A membrane permeable to certain molecules around the molecule interaction zone increases specific molecule nanosensor selectivity response.

  11. Nanodevices for generating power from molecules and batteryless sensing

    Wang, Yinmin; Wang, Xianying; Hamza, Alex V.

    2014-07-15

    A nanoconverter or nanosensor is disclosed capable of directly generating electricity through physisorption interactions with molecules that are dipole containing organic species in a molecule interaction zone. High surface-to-volume ratio semiconductor nanowires or nanotubes (such as ZnO, silicon, carbon, etc.) are grown either aligned or randomly-aligned on a substrate. Epoxy or other nonconductive polymers are used to seal portions of the nanowires or nanotubes to create molecule noninteraction zones. By correlating certain molecule species to voltages generated, a nanosensor may quickly identify which species is detected. Nanoconverters in a series parallel arrangement may be constructed in planar, stacked, or rolled arrays to supply power to nano- and micro-devices without use of external batteries. In some cases breath, from human or other life forms, contain sufficient molecules to power a nanoconverter. A membrane permeable to certain molecules around the molecule interaction zone increases specific molecule nanosensor selectivity response.

  12. Hadron Molecules

    Gutsche, Thomas; Faessler, Amand; Lee, Ian Woo; Lyubovitskij, Valery E

    2010-01-01

    We discuss a possible interpretation of the open charm mesons $D_{s0}^*(2317)$, $D_{s1}(2460)$ and the hidden charm mesons X(3872), Y(3940) and Y(4140) as hadron molecules. Using a phenomenological Lagrangian approach we review the strong and radiative decays of the $D_{s0}^* (2317)$ and $D_{s1}(2460)$ states. The X(3872) is assumed to consist dominantly of molecular hadronic components with an additional small admixture of a charmonium configuration. Determing the radiative ($\\gamma J/\\psi$ and $\\gamma \\psi(2s)$) and strong ($J/\\psi 2\\pi $ and $ J/\\psi 3\\pi$) decay modes we show that present experimental observation is consistent with the molecular structure assumption of the X(3872). Finally we give evidence for molecular interpretations of the Y(3940) and Y(4140) related to the observed strong decay modes $J/\\psi + \\omega$ or $J/\\psi + \\phi$, respectively.

  13. The PH Domain of PDK1 Exhibits a Novel, Phospho-Regulated Monomer-Dimer Equilibrium With Important Implications for Kinase Domain Activation: Single Molecule and Ensemble Studies†

    Ziemba, Brian P.; Pilling, Carissa; Calleja, Véronique; Larijani, Banafshé; Falke, Joseph J.

    2013-01-01

    Phosphoinositide-Dependent Kinase-1 (PDK1) is an essential master kinase recruited to the plasma membrane by the binding of its C-terminal PH domain to the signaling lipid phosphatidylinositol-3,4-5-trisphosphate (PIP3). Membrane binding leads to PDK1 phospho-activation, but despite the central role of PDK1 in signaling and cancer biology this activation mechanism remains poorly understood. PDK1 has been shown to exist as a dimer in cells, and one crystal structure of its isolated PH domain exhibits a putative dimer interface. It has been proposed that phosphorylation of PH domain residue T513 (or the phospho-mimetic T513E mutation) may regulate a novel PH domain dimer-monomer equilibrium, thereby converting an inactive PDK1 dimer to an active monomer. However, the oligomeric state(s) of the PH domain on the membrane have not yet been determined, nor whether a negative charge at position 513 is sufficient to regulate its oligomeric state. The present study investigates the binding of purified WT and T513E PDK1 PH domains to lipid bilayers containing the PIP3 target lipid, using both single molecule and ensemble measurements. Single molecule analysis of the brightness of fluorescent PH domain shows that the PIP3-bound WT PH domain on membranes is predominantly dimeric, while the PIP3-bound T513E PH domain is monomeric, demonstrating that negative charge at the T513 position is sufficient to dissociate the PH domain dimer and is thus likely to play a central role in PDK1 monomerization and activation. Single molecule analysis of 2-D diffusion of PH domain-PIP3 complexes reveals that the dimeric WT PH domain diffuses at the same rate a single lipid molecule, indicating that only one of its two PIP3 binding sites is occupied and there is little protein penetration into the bilayer as observed for other PH domains. The 2-D diffusion of T513E PH domain is slower, suggesting the negative charge disrupts local structure in a way that enables greater protein insertion into

  14. Neohesperidin dihydrochalcone: presentation of a small molecule activator of mammalian alpha-amylase as an allosteric effector.

    Kashani-Amin, Elaheh; Larijani, Bagher; Ebrahim-Habibi, Azadeh

    2013-03-18

    Flavonoids and their precursor trans-chalcone have been reported as inhibitors of mammalian alpha-amylase. With regard to this background, neohesperidin dihydrochalcone (NHDC) effect was investigated toward porcine pancreatic alpha-amylase (PPA), and found to be an activator of the enzyme. The maximal activation (up to threefold) was found to occur at 4.8mM of NHDC, which could be considered to have a high activation profile, with regard to the alpha and beta parameters (alpha<1activator of the enzyme and based on the results obtained from modeling tools, it is suggested to interact with PPA at a hydrophilic site located at the N-terminal, far from the active site of the enzyme.

  15. Atoms, molecules & elements

    Graybill, George

    2007-01-01

    Young scientists will be thrilled to explore the invisible world of atoms, molecules and elements. Our resource provides ready-to-use information and activities for remedial students using simplified language and vocabulary. Students will label each part of the atom, learn what compounds are, and explore the patterns in the periodic table of elements to find calcium (Ca), chlorine (Cl), and helium (He) through hands-on activities.

  16. Gated container molecules

    LIU Fang; WANG Hao; HOUK K. N.

    2011-01-01

    Donald J.Cram,the great UCLA chemist,received the Nobel Prize for his discoveries about host-guest complexes [1].Both theoretical and experimental studies have been conducted about the nature and strength of interactions between the host and guest molecules.The concepts of constrictive binding (the activation energy of the binding process) and intrinsic binding (the free energy difference between the complex and the free host and guest molecules) were introduced to characterize different binding properties (Figure 1)[2].

  17. Sequential activation of p75 and TrkB is involved in dendritic development of subventricular zone-derived neuronal progenitors in vitro.

    Gascon, E; Vutskits, L; Zhang, H; Barral-Moran, M J; Kiss, P J; Mas, C; Kiss, J Z

    2005-01-01

    Dendritic arbor development of subventricular zone-derived interneurons is a critical step in their integration into functional circuits of the postnatal olfactory bulb. However, the mechanism and molecular control of this process remain unknown. In this study, we have developed a culture model where dendritic development of purified subventricular zone cells proceeds under serum-free conditions in the absence of added growth factors and non-neural cells. We demonstrate that the large majority of these cells in culture express GABA and elaborate dendritic arbors with spine-like protrusions but they do not possess axons. These neurons expressed receptors for neurotrophins including p75, TrkB and TrkC but not TrkA. Application of exogenous neurotrophins, including brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT3) and nerve growth factor (NGF), to cultures stimulated dendritic growth and led to more complex dendritic arbors during the initial 3 days in culture. Our results suggest that these effects are independent of Trk receptors and mediated by the p75/ceramide signaling pathway. We also show that brain-derived neurotrophic factor is the only neurotrophin that is able to influence late-phase dendritic development via TrkB receptor activation. These results suggest that dendritic arbor development of subventricular zone-derived cells may be regulated by neurotrophins through the activation of p75 and the TrkB receptor signaling pathways in a sequentially defined temporal pattern.

  18. Proteolysis of neuronal cell adhesion molecule by the tissue plasminogen activator-plasmin system after kainate injection in the mouse hippocampus.

    Endo, A; Nagai, N; Urano, T; Takada, Y; Hashimoto, K; Takada, A

    1999-01-01

    Tissue plasminogen activator (tPA) is a serine protease that converts inactive plasminogen to the active protease plasmin and mediates extracellular metabolism. tPA is transcriptionally induced in the mouse hippocampus by pharmacological or electrical stimulation of neuronal activity and mediates excitotoxin-induced neuronal degeneration. Therefore, we hypothesized that tPA would be induced in the hippocampus after kainic acid (KA) injection into the lateral cerebral ventricle (LCV) and that the activated tPA-plasmin system would degrade the neuronal cell adhesion molecule (NCAM), which is a component of the extracellular matrix. In order to investigate this possibility, we first examined whether NCAM is a substrate for the tPA plasmin system by incubating mouse brain homogenates with tPA and plasminogen at 37 degrees C. Next, we examined the degradation of NCAM and the changes of tPA activity in the mouse hippocampus with immunohistochemical procedures and histological zymography after KA injection into both LCVs. As a result, we observed neuronal atrophy and a decrease of NCAM immunoreactivity along with an increase of tPA activity in the CA3 area of the hippocampus. These results suggest that activation of the tPA plasmin system after KA injection into the LCVs results in the degradation of NCAM in the CA3 area.

  19. Hybrid Molecules Containing a 7-Chloro-4-aminoquinoline Nucleus and a Substituted 2-Pyrazoline with Antiproliferative and Antifungal Activity

    Alba Montoya

    2016-07-01

    Full Text Available Twenty-four new hybrid analogues (15–38 containing 7-chloro-4-aminoquinoline and 2-pyrazoline N-heterocyclic fragments were synthesized. Twelve of the new compounds were evaluated against 58 human cancer cell lines by the U.S. National Cancer Institute (NCI. Compounds 25, 30, 31, 36, and 37 showed significant cytostatic activity, with the most outstanding GI50 values ranging from 0.05 to 0.95 µM. The hybrid compounds (15–38 were also evaluated for antifungal activity against Candida albicans and Cryptococcus neoformans. From the obtained results some structure–activity relationships were outlined.

  20. Hybrid Molecules Containing a 7-Chloro-4-aminoquinoline Nucleus and a Substituted 2-Pyrazoline with Antiproliferative and Antifungal Activity.

    Montoya, Alba; Quiroga, Jairo; Abonia, Rodrigo; Derita, Marcos; Sortino, Maximiliano; Ornelas, Alfredo; Zacchino, Susana; Insuasty, Braulio

    2016-07-27

    Twenty-four new hybrid analogues (15-38) containing 7-chloro-4-aminoquinoline and 2-pyrazoline N-heterocyclic fragments were synthesized. Twelve of the new compounds were evaluated against 58 human cancer cell lines by the U.S. National Cancer Institute (NCI). Compounds 25, 30, 31, 36, and 37 showed significant cytostatic activity, with the most outstanding GI50 values ranging from 0.05 to 0.95 µM. The hybrid compounds (15-38) were also evaluated for antifungal activity against Candida albicans and Cryptococcus neoformans. From the obtained results some structure-activity relationships were outlined.

  1. Plant-derived flavone Apigenin: The small-molecule with promising activity against therapeutically resistant prostate cancer.

    Ganai, Shabir Ahmad

    2017-01-01

    Prostate cancer is the second leading cause of cancer related deaths in men in the United States. Mounting evidences suggest that in the pathophysiology of prostate cancer epigenetic modifications play a considerable role. Histone deacetylases (HDACs) have strong crosstalk with prostate cancer progression as they regulate various genes meant for tumour suppression. HDACs are emerging as striking molecular targets for anticancer drugs and therapy as their aberrant expression has been implicated in several cancers. Histone deacetylase inhibitors (HDACi), the small molecules interfering HDACs are the propitious chemotherapeutic agents as they tune the altered acetylation homeostasis for attenuating disease signalling. More than 20 HDACi have entered into the clinical trials and 4 have crossed the journey by gaining FDA approval for treating distinct haematological malignancies including multiple myeloma. Despite the therapeutic benefits, the synthetic HDACi cause detrimental side effects like atrial fibrillation, raising concerns regarding their applicability. Taking these facts into consideration the current article focused on plant-derived HDAC inhibitor Apigenin and its marvelous role in prostate cancer therapy. Moreover, the article sheds light on Apigenin induced apoptosis in various prostate cancer models. The defined inhibitor provokes apoptotic signaling in these models by multiple mechanisms like restraining HDACs, declining the levels of antiapoptotic proteins. Importantly, Apigenin hampers NF-κB signalling and down-modulates its regulated gene products for bringing therapeutic effect. Furthermore, Apigenin shows synergistic effect in combinatorial therapy and induces apoptosis even in prostate cancer models resistant to conventional therapeutic regimens.

  2. Current strategies for targeted delivery of bio-active drug molecules in the treatment of brain tumor.

    Garg, Tarun; Bhandari, Saurav; Rath, Goutam; Goyal, Amit K

    2015-12-01

    Brain tumor is one of the most challenging diseases to treat. The major obstacle in the specific drug delivery to brain is blood-brain barrier (BBB). Mostly available anti-cancer drugs are large hydrophobic molecules which have limited permeability via BBB. Therefore, it is clear that the protective barriers confining the passage of the foreign particles into the brain are the main impediment for the brain drug delivery. Hence, the major challenge in drug development and delivery for the neurological diseases is to design non-invasive nanocarrier systems that can assist controlled and targeted drug delivery to the specific regions of the brain. In this review article, our major focus to treat brain tumor by study numerous strategies includes intracerebral implants, BBB disruption, intraventricular infusion, convection-enhanced delivery, intra-arterial drug delivery, intrathecal drug delivery, injection, catheters, pumps, microdialysis, RNA interference, antisense therapy, gene therapy, monoclonal/cationic antibodies conjugate, endogenous transporters, lipophilic analogues, prodrugs, efflux transporters, direct conjugation of antitumor drugs, direct targeting of liposomes, nanoparticles, solid-lipid nanoparticles, polymeric micelles, dendrimers and albumin-based drug carriers.

  3. Synthesis and characterization of perm-selective SERS-active silica-coated gold nanospheres for the direct detection of small molecules

    Pierre-Bolivar, Marie Carmelle Serviane

    Noble metal nanomaterials have numerous uses in plasmonic and surface enhanced Raman scattering (SERS) detection applications; however, upon the addition of analytes, nanomaterials often undergo uncontrolled aggregation which leads to inconsistent signal intensities. To overcome this limitation, the effect of gold nanosphere concentration, column purification, and surface chemistry functionalization using internally etched silica stabilization methods was investigated on SERS assays for small molecule detection. Nanostructure composition, size, shape, stability, surface chemistry, optical properties, and SERS-activity were monitored using localized surface plasmon resonance (LSPR or extinction) spectroscopy, transmission electron microscopy (TEM), and Raman spectroscopy. First, the behavior of citrate-stabilized gold nanospheres was monitored as a function of molecular surface coverage. Both extinction and SERS spectral intensities increased linearly below monolayer functionalization. Above this value, however, uncontrolled nanoparticle aggregation occurred and large but irreproducible SERS signal intensities were monitored. Next, gold nanoparticles were encapsulated with varying silica shell thicknesses and purified using traditional centrifugation steps and/or column chromatography. Relative to the traditionally purified (i.e. centrifuged) samples, the SERS responses from small molecules using the column purified nanoparticle samples followed a well-known SERS distance-dependence model. Thus, surface chemistry cannot form more than a 2 nm thick layer on gold nanospheres if SERS applications were targeted. To overcome these challenges, gold nanospheres encapsulated with a thick silica shell were made SERS-active by etching the internal silica layer near the metal surface. During the synthesis of these internally etched silica-coated gold nanospheres, the LSPR wavelength shift, a parameter related to the effective local refractive index near the gold core, was

  4. Structural and Functional Characterization of a Single-chain Peptide-MHC Molecule that Modulates both Naive and Activated CD8plus T Cells

    D Samanta; G Mukherjee; U Ramagopal; R Chaparro; S Nathenson; T DiLorenzo; S Almo

    2011-12-31

    Peptide-MHC (pMHC) multimers, in addition to being tools for tracking and quantifying antigen-specific T cells, can mediate downstream signaling after T-cell receptor engagement. In the absence of costimulation, this can lead to anergy or apoptosis of cognate T cells, a property that could be exploited in the setting of autoimmune disease. Most studies with class I pMHC multimers used noncovalently linked peptides, which can allow unwanted CD8{sup +} T-cell activation as a result of peptide transfer to cellular MHC molecules. To circumvent this problem, and given the role of self-reactive CD8{sup +} T cells in the development of type 1 diabetes, we designed a single-chain pMHC complex (scK{sup d}.IGRP) by using the class I MHC molecule H-2K{sup d} and a covalently linked peptide derived from islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP{sub 206-214}), a well established autoantigen in NOD mice. X-ray diffraction studies revealed that the peptide is presented in the groove of the MHC molecule in canonical fashion, and it was also demonstrated that scK{sup d}.IGRP tetramers bound specifically to cognate CD8{sup +} T cells. Tetramer binding induced death of naive T cells and in vitro- and in vivo-differentiated cytotoxic T lymphocytes, and tetramer-treated cytotoxic T lymphocytes showed a diminished IFN-{gamma} response to antigen stimulation. Tetramer accessibility to disease-relevant T cells in vivo was also demonstrated. Our study suggests the potential of single-chain pMHC tetramers as possible therapeutic agents in autoimmune disease. Their ability to affect the fate of naive and activated CD8{sup +} T cells makes them a potential intervention strategy in early and late stages of disease.

  5. HIV-1 infection ex vivo accelerates measles virus infection by upregulating signaling lymphocytic activation molecule (SLAM) in CD4+ T cells.

    Mitsuki, Yu-ya; Terahara, Kazutaka; Shibusawa, Kentaro; Yamamoto, Takuya; Tsuchiya, Takatsugu; Mizukoshi, Fuminori; Ishige, Masayuki; Okada, Seiji; Kobayashi, Kazuo; Morikawa, Yuko; Nakayama, Tetsuo; Takeda, Makoto; Yanagi, Yusuke; Tsunetsugu-Yokota, Yasuko

    2012-07-01

    Measles virus (MV) infection in children harboring human immunodeficiency virus type 1 (HIV-1) is often fatal, even in the presence of neutralizing antibodies; however, the underlying mechanisms are unclear. Therefore, the aim of the present study was to examine the interaction between HIV-1 and wild-type MV (MVwt) or an MV vaccine strain (MVvac) during dual infection. The results showed that the frequencies of MVwt- and MVvac-infected CD4(+) T cells within the resting peripheral blood mononuclear cells (PBMCs) were increased 3- to 4-fold after HIV-1 infection, and this was associated with a marked upregulation of signaling lymphocytic activation molecule (SLAM) expression on CD4(+) T cells but not on CD8(+) T cells. SLAM upregulation was induced by infection with a replication-competent HIV-1 isolate comprising both the X4 and R5 types and to a lesser extent by a pseudotyped HIV-1 infection. Notably, SLAM upregulation was observed in HIV-infected as well as -uninfected CD4(+) T cells and was abrogated by the removal of HLA-DR(+) cells from the PBMC culture. Furthermore, SLAM upregulation did not occur in uninfected PBMCs cultured together with HIV-infected PBMCs in compartments separated by a permeable membrane, indicating that no soluble factors were involved. Rather, CD4(+) T cell activation mediated through direct contact with dendritic cells via leukocyte function-associated molecule 1 (LFA-1)/intercellular adhesion molecule 1 (ICAM-1) and LFA-3/CD2 was critical. Thus, HIV-1 infection induces a high level of SLAM expression on CD4(+) T cells, which may enhance their susceptibility to MV and exacerbate measles in coinfected individuals.

  6. Activating the Zone of Proximal Development of Japanese Language Learners: Language-Exchange Partnerships (LEPs) at an Australian University

    Hiromi Nishioka

    2014-01-01

    This study examines how social interactions in language-exchange partnership sessions can create opportunities for Japanese language learners to use and learn Japanese. The participants in this study were two pairs, consisting of a Japanese native speaker and an Australian studying Japanese at an Australian university. This study, employing Vygotsky’s Zone of Proximal Development and assistance to novices by experts, examined what factors enable participants in language-exchange partnerships ...

  7. The Role of Sulfhydryl Reactivity of Small Molecules for the Activation of the KEAP1/NRF2 Pathway and the Heat Shock Response

    Albena T. Dinkova-Kostova

    2012-01-01

    Full Text Available The KEAP1/NRF2 pathway and the heat shock response are two essential cytoprotective mechanisms that allow adaptation and survival under conditions of oxidative, electrophilic, and thermal stress by regulating the expression of elaborate networks of genes with versatile protective functions. The two pathways are independently regulated by the transcription factor nuclear factor-erythroid 2 p45-related factor 2 (NRF2 and heat shock factor 1 (HSF1, respectively. The activity of these transcriptional master regulators increases during conditions of stress and also upon encounter of small molecules (inducers, both naturally occurring as well as synthetically produced. Inducers have a common chemical property: the ability to react with sulfhydryl groups. The protein targets of such sulfhydryl-reactive compounds are equipped with highly reactive cysteine residues, which serve as sensors for inducers. The initial cysteine-sensed signal is further relayed to affect the expression of large networks of genes, which in turn can ultimately influence complex cell fate decisions such as life and death. The paper summarizes the multiple lines of experimental evidence demonstrating that the reactivity with sulfhydryl groups is a major determinant of the mechanism of action of small molecule dual activators of the KEAP1/NRF2 pathway and the heat shock response.

  8. Pollution Status of Trace Metals in Groundwater Due to Industrail Activities in and Around Dhaka Export Processing Zone, Bangladesh

    GOLAM AHMED

    2012-06-01

    Full Text Available Effluents from multiindustrail activities influence inland water system directly, which subsiquently affect groundwater quality and human health. Some previous reports indicated that inadequate treatment process of discharged effluent of Dhaka Export Processing Zone (DEPZ increased the concentrations of pollutants in surface water system and deteriorated total fishing and agricultural system around DEPZ and its connected area. Therefore, the present study was conducted to investigate wether the concentration of selective metals viz. Li, V, Cr, Co, Ni, Cu, Zn, Ga, As, Se, Rb, Sr, Ag, Cd, Cs, Ba, Pb and U in two types of groundwater sources were either with in the permissible guidlines or influenced by DEPZ multi industrail on their levels of contamination. The concentrations of metals were determined using inductively Couples Plasma Mass Spectrometry (ICP-MS. The mean concentrations of the elements in both types of groundwater were in the levels of their permissible guidlines, except for Ni (12.91 µg/L, Ga (0.48µg/L, Sr (90.26 µg/L and Cs (0.07µg//Lin groundwater inside DEPZ, which were 1.30, 5.00, 1.50 and 1.40 times higher than the maximum permissible limit (MPL of 10 µg/L, 0.09 µg/L, 60 µg/L, and 0.05µg/L, respectively. The mean concentrations of Li (6.85 µg/L, Zn(268 µg/L, Ga (0.12 µg/L, Sr (131 µg/L and Cs (0.07 µg/L were 3.43, 1.34, 1.33, 2.18, 1.40 times higher then the MPL of 2 µg/L, 200 µg/L, 0.09 µg/L, 60 µg/L and 0.05 µg/L, respectively, in groundwater around DEPZ. Comparatively Zn and Sr possessed higher concentrations, and Cs and U possessed lower concentration in both types of groundwater sources. The elements were distributed in homogeneous and hetrogeneous manner among the source points for deep-tubewell (DTWS and shallow tubewell (STWs, respectively. The significant positive correlations were found between the elements viz., Co-V (0.85, Ni-Sr ((0.70, Co-Cd (0.86, As-Se (0.99, Cs-Zn (0.95, Li-U (0.,71, Zn-U (0

  9. The Autophagy Molecule Beclin 1 Maintains Persistent Activity of NF-κB and Stat3 in HTLV-1-transformed T Lymphocytes

    Chen, Li; Liu, Dan; Zhang, Yang; Zhang, Huan; Cheng, Hua

    2015-01-01

    The retroviral oncoprotein Tax from human T cell leukemia virus type 1 (HTLV-1) induces persistent activation of IκB kinase (IKK)/NF-κB signaling, an essential step for initiating HTLV-1 oncogenesis. The regulation of the IKK/NF-κB signaling in HTLV-1-transformed T cells remains incompletely understood. In the present study, we showed that the autophagy molecule Beclin1 not only executed a cytoprotective function through induction of autophagy but also played a pivotal role in maintaining Tax-induced activation of two key survival factors, NF-κB and Stat3. Silencing Beclin1 in HTLV-1-transformed T cells resulted in diminished activities of NF-κB and Stat3 as well as impaired growth. In Beclin1-depleted cells, Tax failed to activate NF-κB and Stat3 at its full capacity. In addition, we showed that Beclin1 interacted with the catalytic subunits of IKK. Further, we observed that selective inhibition of IKK repressed the activities of both NF-κB and Stat3 in the context of HTLV-1-transformation of T cells. Our data, therefore, unveiled a key role of Beclin1 in maintaining persistent activities of both NF-κB and Stat3 in the pathogenesis of HTLV-1-mediated oncogenesis. PMID:26319552

  10. Novel metal chelating molecules with anticancer activity. Striking effect of the imidazole substitution of the histidine-pyridine-histidine system.

    Ali, Taha F S; Iwamaru, Kana; Ciftci, Halil Ibrahim; Koga, Ryoko; Matsumoto, Masahiro; Oba, Yasunori; Kurosaki, Hiromasa; Fujita, Mikako; Okamoto, Yoshinari; Umezawa, Kazuo; Nakao, Mitsuyoshi; Hide, Takuichiro; Makino, Keishi; Kuratsu, Jun-ichi; Abdel-Aziz, Mohamed; Abuo-Rahma, Gamal El-Din A A; Beshr, Eman A M; Otsuka, Masami

    2015-09-01

    Previously we have reported a metal chelating histidine-pyridine-histidine system possessing a trityl group on the histidine imidazole, namely HPH-2Trt, which induces apoptosis in human pancreatic adenocarcinoma AsPC-1 cells. Herein the influence of the imidazole substitution of HPH-2Trt was examined. Five related compounds, HPH-1Trt, HPH-2Bzl, HPH-1Bzl, HPH-2Me, and HPH-1Me were newly synthesized and screened for their activity against AsPC-1 and brain tumor cells U87 and U251. HPH-1Trt and HPH-2Trt were highly active among the tested HPH compounds. In vitro DNA cleavage assay showed both HPH-1Trt and HPH-2Trt completely disintegrate pUC19 DNA. The introduction of trityl group decisively potentiated the activity.

  11. Small molecules CK-666 and CK-869 inhibit actin-related protein 2/3 complex by blocking an activating conformational change.

    Hetrick, Byron; Han, Min Suk; Helgeson, Luke A; Nolen, Brad J

    2013-05-23

    Actin-related protein 2/3 (Arp2/3) complex is a seven-subunit assembly that nucleates branched actin filaments. Small molecule inhibitors CK-666 and CK-869 bind to Arp2/3 complex and inhibit nucleation, but their modes of action are unknown. Here, we use biochemical and structural methods to determine the mechanism of each inhibitor. Our data indicate that CK-666 stabilizes the inactive state of the complex, blocking movement of the Arp2 and Arp3 subunits into the activated filament-like (short pitch) conformation, while CK-869 binds to a serendipitous pocket on Arp3 and allosterically destabilizes the short pitch Arp3-Arp2 interface. These results provide key insights into the relationship between conformation and activity in Arp2/3 complex and will be critical for interpreting the influence of the inhibitors on actin filament networks in vivo.

  12. Enzymatic activities and arbuscular mycorrhizal colonization of Plantago lanceolata and Plantago major in a soil root zone under heavy metal stress.

    Gucwa-Przepióra, Ewa; Nadgórska-Socha, Aleksandra; Fojcik, Barbara; Chmura, Damian

    2016-03-01

    The objectives of the present field study were to examine the soil enzyme activities in the soil root zones of Plantago lanceolata and Plantago major in different heavy metal contaminated stands. Moreover, the investigations concerned the intensity of root endophytic colonization and metal bioaccumulation in roots and shoots. The investigated Plantago species exhibited an excluder strategy, accumulating higher metal content in the roots than in the shoots. The heavy metal accumulation levels found in the two plantain species in this study were comparable to other plants suggested as phytostabilizers; therefore, the selected Plantago species may be applied in the phytostabilization of heavy metal contaminated areas. The lower level of soil enzymes (dehydrogenase, urease, acid, and alkaline phosphatase) as well as the higher bioavailability of metals in the root zone soil of the two plantain species were found in an area affected by smelting activity, where organic matter content in the soil was also the smallest. Mycorrhizal colonization on both species in the contaminated area was similar to colonization in non-contaminated stands. However, the lowest arbuscule occurrence and an absence of dark septate endophytes were found in the area affected by the smelting activity. It corresponded with the lowest plant cover observed in this stand. The assessment of enzyme activity, mycorrhizal colonization, and the chemical and physical properties of soils proved to be sensitive to differences between sites and between Plantago species.

  13. Magnetically separable nanocomposites with photocatalytic activity under visible light for the selective transformation of biomass-derived platform molecules

    Novel magnetically separable TiO2-guanidine-(Ni,Co)Fe2O4 nanomaterials were prepared and characterised by a series of techniques including XRD, SEM, TEM, N2 physisorption as well as XPS and subsequently tested for their photocatalytic activities in the selective transformation of...

  14. Teaching with the Case Study Method to Promote Active Learning in a Small Molecule Crystallography Course for Chemistry Students

    Campbell, Michael G.; Powers, Tamara M.; Zheng, Shao-Liang

    2016-01-01

    Implementing the case study method in a practical X-ray crystallography course designed for graduate or upper-level undergraduate chemistry students is described. Compared with a traditional lecture format, assigning small groups of students to examine literature case studies encourages more active engagement with the course material and…

  15. Same molecule but different expression: aging and sepsis trigger NLRP3 inflammasome activation, a target of melatonin.

    Volt, Huayqui; García, José A; Doerrier, Carolina; Díaz-Casado, María E; Guerra-Librero, Ana; López, Luis C; Escames, Germaine; Tresguerres, Jesús A; Acuña-Castroviejo, Darío

    2016-03-01

    The connection between the innate immune system, clock genes, and mitochondrial bioenergetics was analyzed during aging and sepsis in mouse heart. Our results suggest that the sole NF-κB activation does not explain the inflammatory process underlying aging; the former also triggers the NLRP3 inflammasome that enhances caspase-1-dependent maturation of IL-1β. In this way, aged mice enter into a vicious cycle as IL-1β further activates the NF-κB/NLRP3 inflammasome link. The origin of NF-κB activation was related to the age-dependent Bmal1/Clock/RORα/Rev-Erbα loop disruption, which lowers NAD(+) levels, reducing the SIRT1 deacetylase ability to inactivate NF-κB. Consequently, NF-κB binding to DNA increases, raising the formation of proinflammatory mediators and inducing mitochondrial impairment. The cycle is then closed with the subsequent NLRP3 inflammasome activation. This paired contribution of the innate immune pathways serves as a catalyst to magnify the response to sepsis in aged compared with young mice. Melatonin administration blunted the septic response, reducing inflammation and oxidative stress, and enhancing mitochondrial function at the levels of nonseptic aged mice, but it did not counteract the age-related inflammation. Together, our results suggest that, although with different strengths, chronoinflammaging constitutes the biochemical substrate of aging and sepsis, and identifies the NLRP3 inflammasome as a new molecular target for melatonin, providing a rationale for its use in NLRP3-dependent diseases.

  16. Hematopoietic lineage cell specific protein 1 (HS1) is a functionally important signaling molecule in platelet activation.

    Kahner, Bryan N; Dorsam, Robert T; Mada, Sripal R; Kim, Soochong; Stalker, Timothy J; Brass, Lawrence F; Daniel, James L; Kitamura, Daisuke; Kunapuli, Satya P

    2007-10-01

    Collagen activates platelets through an intracellular signaling cascade downstream of glycoprotein VI (GPVI). We have investigated the contribution of hematopoietic lineage cell-specific protein 1 (HS1) downstream of GPVI in platelet activation. Stimulation of GPVI leads to tyrosine phosphorylation of HS1, which is blocked by Src-family kinase inhibitors. Coimmunoprecipitation experiments revealed that HS1 associates with Syk and phosphatidylinositol 3-kinases. HS1-null mice displayed increased bleeding times and increased time to occlusion in the FeCl(3) in vivo thrombosis model compared with their wild-type littermates. In addition, aggregation and secretion responses were diminished in HS1-null mouse platelets after stimulation of GPVI and protease-activated receptor 4 (PAR-4) agonists compared with wild-type littermate mouse platelets. Finally, Akt phosphorylation was diminished after GPVI or PAR-4 stimulation in platelets from HS1-null mice compared with their wild-type littermates. These results demonstrate that phosphorylation of the HS1 protein occurs downstream of GPVI stimulation and that HS1 plays a significant functional role in platelet activation downstream of GPVI and PARs.

  17. The active sequence in the acth molecule responsible for inhibition of the extinction of conditioned avoidance behaviour in rats

    Greven, H.M.; Wied, D. de

    1967-01-01

    The effect of structural analogues of the N-terminal decapeptide of ACTH on inhibition of extinction of a conditioned avoidance response in rats has been studied. Studies involving the relation between chain length and behavioural activity revealed that the sequence 4–10 is the shortest peptide whic

  18. Implementation of a high-throughput screen for identifying small molecules to activate the Keap1-Nrf2-ARE pathway.

    Kai Connie Wu

    Full Text Available Nuclear factor erythroid 2-related factor 2 (Nrf2 is a transcription factor that induces a battery of cytoprotective genes involved in antioxidant defense through binding to Antioxidant Response Elements (ARE located in the promoter regions of these genes. To identify Nrf2 activators for the treatment of oxidative/electrophilic stress-induced diseases, the present study developed a high-throughput assay to evaluate Nrf2 activation using AREc32 cells that contain a luciferase gene under the control of ARE promoters. Of the 47,000 compounds screened, 238 (top 0.5% hits of the chemicals increased the luminescent signal more than 14.4-fold and were re-tested at eleven concentrations in a range of 0.01-30 µM. Of these 238 compounds, 231 (96% increased the luminescence signal in a concentration-dependent manner. Chemical structure relationship analysis of these 231 compounds indicated enrichment of four chemical scaffolds (diaryl amides and diaryl ureas, oxazoles and thiazoles, pyranones and thiapyranones, and pyridinones and pyridazinones. In addition, 30 of these 231 compounds were highly effective and/or potent in activating Nrf2, with a greater than 80-fold increase in luminescence, or an EC50 lower than 1.6 µM. These top 30 compounds were also screened in Hepa1c1c7 cells for an increase in Nqo1 mRNA, the prototypical Nrf2-target gene. Of these 30 compounds, 17 increased Nqo1 mRNA in a concentration-dependent manner. In conclusion, the present study documents the development, implementation, and validation of a high-throughput screen to identify activators of the Keap1-Nrf2-ARE pathway. Results from this screening identified Nrf2 activators, and provide novel insights into chemical scaffolds that might prevent oxidative/electrophilic stress-induced toxicity and carcinogenesis.

  19. The northwest trending north Boquerón Bay-Punta Montalva Fault Zone; A through going active fault system in southwestern Puerto Rico

    Roig‐Silva, Coral Marie; Asencio, Eugenio; Joyce, James

    2013-01-01

    The North Boquerón Bay–Punta Montalva fault zone has been mapped crossing the Lajas Valley in southwest Puerto Rico. Identification of the fault was based upon detailed analysis of geophysical data, satellite images, and field mapping. The fault zone consists of a series of Cretaceous bedrock faults that reactivated and deformed Miocene limestone and Quaternary alluvial fan sediments. The fault zone is seismically active (local magnitude greater than 5.0) with numerous locally felt earthquakes. Focal mechanism solutions suggest strain partitioning with predominantly east–west left-lateral displacements with small normal faults striking mostly toward the northeast. Northeast-trending fractures and normal faults can be found in intermittent streams that cut through the Quaternary alluvial fan deposits along the southern margin of the Lajas Valley, an east–west-trending 30-km-long fault-controlled depression. Areas of preferred erosion within the alluvial fan trend toward the west-northwest parallel to the onland projection of the North Boquerón Bay fault. The North Boquerón Bay fault aligns with the Punta Montalva fault southeast of the Lajas Valley. Both faults show strong southward tilting of Miocene strata. On the western end, the Northern Boquerón Bay fault is covered with flat-lying Holocene sediments, whereas at the southern end the Punta Montalva fault shows left-lateral displacement of stream drainage on the order of a few hundred meters.

  20. Molecule modification and antioxidant activity of EGC%EGC分子修饰及其抗氧化活性

    刘金鹏; 蔡烈伟; 杜琪珍

    2012-01-01

    亲水性的表没食子儿茶素(ECC)是绿茶茶多酚的组成成分之一,采用氧杂Pictet-Spengler反应,通过嫁接酮类物质对表没食子儿茶素(EGC)进行了分子修饰,并对反应产物进行了抗氧化活性的研究。反应产物为脂溶性成分,且保持了较高的抗氧化活性。%Water-soluble EGC is one of the composition of green tea polyphenols. In this paper, Oxa- Pictet-Spengler reaction is use to modify EGC by grafting ketones, and studying the antioxidant activity of products. Products is liposoluble components, retaining fine antioxidant activity.

  1. Enzyme kinetics and distinct modulation of the protein kinase N family of kinases by lipid activators and small molecule inhibitors

    Falk, Matthew D.; Liu, Wei; Bolaños, Ben; Unsal-Kacmaz, Keziban; Klippel, Anke; Grant, Stephan; Brooun, Alexei; Timofeevski, Sergei

    2014-01-01

    The PKN (protein kinase N) family of Ser/Thr protein kinases regulates a diverse set of cellular functions, such as cell migration and cytoskeletal organization. Inhibition of tumour PKN activity has been explored as an oncology therapeutic approach, with a PKN3-targeted RNAi (RNA interference)-derived therapeutic agent in Phase I clinical trials. To better understand this important family of kinases, we performed detailed enzymatic characterization, determining the kinetic mechanism and lipi...

  2. New Perspectives on Specific Immune-Depletion Technique Using Monoclonal Antibodies against Small Active Molecules in Herbs

    2014-01-01

    One of the main focuses in Chinese Medicine research is the identification of efficacious components in Chinese herbal medicine (CHM). Studies in such area are difficult due to the complexity and the synergistic characteristics of CHM. Current methods to track and separate active components are not adequate to meet the needs of revealing effects and identify substances and pharmacological mechanisms, which directly restrict the modernization and globalization of CHM. In this paper, a new meth...

  3. Small molecule binding sites on the Ras:SOS complex can be exploited for inhibition of Ras activation.

    Winter, Jon J G; Anderson, Malcolm; Blades, Kevin; Brassington, Claire; Breeze, Alexander L; Chresta, Christine; Embrey, Kevin; Fairley, Gary; Faulder, Paul; Finlay, M Raymond V; Kettle, Jason G; Nowak, Thorsten; Overman, Ross; Patel, S Joe; Perkins, Paula; Spadola, Loredana; Tart, Jonathan; Tucker, Julie A; Wrigley, Gail

    2015-03-12

    Constitutively active mutant KRas displays a reduced rate of GTP hydrolysis via both intrinsic and GTPase-activating protein-catalyzed mechanisms, resulting in the perpetual activation of Ras pathways. We describe a fragment screening campaign using X-ray crystallography that led to the discovery of three fragment binding sites on the Ras:SOS complex. The identification of tool compounds binding at each of these sites allowed exploration of two new approaches to Ras pathway inhibition by stabilizing or covalently modifying the Ras:SOS complex to prevent the reloading of Ras with GTP. Initially, we identified ligands that bound reversibly to the Ras:SOS complex in two distinct sites, but these compounds were not sufficiently potent inhibitors to validate our stabilization hypothesis. We conclude by demonstrating that covalent modification of Cys118 on Ras leads to a novel mechanism of inhibition of the SOS-mediated interaction between Ras and Raf and is effective at inhibiting the exchange of labeled GDP in both mutant (G12C and G12V) and wild type Ras.

  4. Vasorelaxant activity of extracts obtained from Apium graveolens:Possible source for vasorelaxant molecules isolation with potential antihypertensive effect

    Vergara-Galicia Jorge; Jimenez-Ramirez Luis ngel; Tun-Suarez Adrin; Aguirre-Crespo Francisco; Salazar-Gmez Anuar; Estrada-Soto Samuel; Sierra-Ovando ngel; Hernandez-Nuez Emmanuel

    2013-01-01

    Objective:To investigate the vasorelaxant effect of organic extracts from Apium graveolens (A. graveolens) which is a part of a group of plants subjected to pharmacological and phytochemical study with the purpose of offering it as an ideal source for obtaining lead compounds for designing new therapeutic agents with potential vasorelaxant and antihypertensive effects. Methods:An ex vivo method was employed to assess the vasorelaxant activity. This consisted of using rat aortic rings with and without endothelium precontracted with norepinephrine. Results:All extracts caused concentration-dependent relaxation in precontracted aortic rings with and without endothelium;the most active extracts were Dichloromethane and Ethyl Acetate extracts from A. graveolens. These results suggested that secondary metabolites responsible for the vasorelaxant activity belong to a group of compounds of medium polarity. Also, our evidence showed that effect induced by dichloromethane and ethyl acetate extracts from A. graveolens is mediated probably by calcium antagonism. Conclusions: A. graveolens represents an ideal source for obtaining lead compounds for designing new therapeutic agents with potential vasorelaxant and antihypertensive effects.

  5. Synthesis and pharmacological evaluation of polyfunctional benzimidazole-NSAID chimeric molecules combining anti-inflammatory, immunomodulatory and antioxidant activities.

    Bansal, Yogita; Silakari, Om

    2014-11-01

    Polyfunctional compounds comprise a novel class of therapeutic agents for treatment of multifactorial diseases. The present study reports a series of benzimidazole-non-steroidal anti-inflammatory drugs (NSAIDs) conjugates (1-10) as novel polyfunctional compounds synthesized in the presence of orthophosphoric acid. The compounds were evaluated for anti-inflammatory (carageenan-induced paw edema model), immunomodulatory (direct haemagglutination test and carbon clearance index models), antioxidant (in vitro and in vivo) and for ulcerogenic effects. Each of the compound has retained the anti-inflammatory activity of the corresponding parent NSAID while exhibiting significantly reduced gastric ulcers. Additionally, the compounds are found to possess potent immunostimulatory and antioxidant activities. The compound 8 was maximally potent (antibody titre value 358.4 ± 140.21, carbon clearance index 0.053 ± 0.002 and antioxidant EC50 value 0.03 ± 0.006). These compounds, exhibiting such multiple pharmacological activities, can be taken as lead for the development of potent drugs for the treatment of chronic multifactorial diseases involving inflammation, immune system modulation and oxidative stress such as cancers. The Lipinski's parameters suggested the compounds to be bear drug like properties.

  6. Scorpionate-type coordination in MFU-4l metal-organic frameworks: small-molecule binding and activation upon the thermally activated formation of open metal sites.

    Denysenko, Dmytro; Grzywa, Maciej; Jelic, Jelena; Reuter, Karsten; Volkmer, Dirk

    2014-06-01

    Postsynthetic metal and ligand exchange is a versatile approach towards functionalized MFU-4l frameworks. Upon thermal treatment of MFU-4l formates, coordinatively strongly unsaturated metal centers, such as zinc(II) hydride or copper(I) species, are generated selectively. Cu(I)-MFU-4l prepared in this way was stable under ambient conditions and showed fully reversible chemisorption of small molecules, such as O2, N2, and H2, with corresponding isosteric heats of adsorption of 53, 42, and 32 kJ mol(-1), respectively, as determined by gas-sorption measurements and confirmed by DFT calculations. Moreover, Cu(I)-MFU-4l formed stable complexes with C2H4 and CO. These complexes were characterized by FTIR spectroscopy. The demonstrated hydride transfer to electrophiles and strong binding of small gas molecules suggests these novel, yet robust, metal-organic frameworks with open metal sites as promising catalytic materials comprising earth-abundant metal elements.

  7. Pharmacological activation of CB2 receptors counteracts the deleterious effect of ethanol on cell proliferation in the main neurogenic zones of the adult rat brain

    Patricia eRivera

    2015-09-01

    Full Text Available Chronic alcohol exposure reduces endocannabinoid activity and disrupts adult neurogenesis in rodents, which results in structural and functional alterations. Cannabinoid receptor agonists promote adult neural progenitor cell (NPC proliferation. We evaluated the protective effects of the selective CB1 receptor agonist ACEA, the selective CB2 receptor agonist JWH133 and the fatty-acid amide-hydrolase (FAAH inhibitor URB597, which enhances endocannabinoid receptor activity, on NPC proliferation in rats with forced consumption of ethanol (10% or sucrose liquid diets for two weeks. We performed immunohistochemical and stereological analyses of cells expressing the mitotic phosphorylation of histone-3 (phospho-H3+ and the replicating cell DNA marker 5-bromo-2’-deoxyuridine (BrdU+ in the main neurogenic zones of adult brain: subgranular zone of dentate gyrus (SGZ, subventricular zone of lateral ventricles (SVZ and hypothalamus. Animals were allowed ad libitum ethanol intake (7.3±1.1 g/kg/day after a controlled isocaloric pair-feeding period of sucrose and alcoholic diets. Alcohol intake reduced the number of BrdU+ cells in SGZ, SVZ and hypothalamus. The treatments (URB597, ACEA, JWH133 exerted a differential increase in alcohol consumption over time, but JWH133 specifically counteracted the deleterious effect of ethanol on NPC proliferation in the SVZ and SGZ, and ACEA reversed this effect in the SGZ only. JWH133 also induced an increased number of BrdU+ cells expressing neuron-specific β3-tubulin in the SVZ and SGZ. These results indicated that the specific activation of CB2 receptors rescued alcohol-induced impaired NPC proliferation, which is a potential clinical interest for the risk of neural damage in alcohol dependence.

  8. Pharmacological activation of CB2 receptors counteracts the deleterious effect of ethanol on cell proliferation in the main neurogenic zones of the adult rat brain

    Rivera, Patricia; Blanco, Eduardo; Bindila, Laura; Alen, Francisco; Vargas, Antonio; Rubio, Leticia; Pavón, Francisco J.; Serrano, Antonia; Lutz, Beat; Rodríguez de Fonseca, Fernando; Suárez, Juan

    2015-01-01

    Chronic alcohol exposure reduces endocannabinoid activity and disrupts adult neurogenesis in rodents, which results in structural and functional alterations. Cannabinoid receptor agonists promote adult neural progenitor cell (NPC) proliferation. We evaluated the protective effects of the selective CB1 receptor agonist ACEA, the selective CB2 receptor agonist JWH133 and the fatty-acid amide-hydrolase (FAAH) inhibitor URB597, which enhances endocannabinoid receptor activity, on NPC proliferation in rats with forced consumption of ethanol (10%) or sucrose liquid diets for 2 weeks. We performed immunohistochemical and stereological analyses of cells expressing the mitotic phosphorylation of histone-3 (phospho-H3+) and the replicating cell DNA marker 5-bromo-2'-deoxyuridine (BrdU+) in the main neurogenic zones of adult brain: subgranular zone of dentate gyrus (SGZ), subventricular zone of lateral ventricles (SVZ) and hypothalamus. Animals were allowed ad libitum ethanol intake (7.3 ± 1.1 g/kg/day) after a controlled isocaloric pair-feeding period of sucrose and alcoholic diets. Alcohol intake reduced the number of BrdU+ cells in SGZ, SVZ, and hypothalamus. The treatments (URB597, ACEA, JWH133) exerted a differential increase in alcohol consumption over time, but JWH133 specifically counteracted the deleterious effect of ethanol on NPC proliferation in the SVZ and SGZ, and ACEA reversed this effect in the SGZ only. JWH133 also induced an increased number of BrdU+ cells expressing neuron-specific β3-tubulin in the SVZ and SGZ. These results indicated that the specific activation of CB2 receptors rescued alcohol-induced impaired NPC proliferation, which is a potential clinical interest for the risk of neural damage in alcohol dependence. PMID:26483633

  9. The adjuvant activity of alphavirus replicons is enhanced by incorporating the microbial molecule flagellin into the replicon.

    Maria L Knudsen

    Full Text Available Ligands of pattern recognition receptors (PRRs including Toll-like receptors (TLRs stimulate innate and adaptive immune responses and are considered as potent adjuvants. Combinations of ligands might act in synergy to induce stronger and broader immune responses compared to stand-alone ligands. Alphaviruses stimulate endosomal TLRs 3, 7 and 8 as well as the cytoplasmic PRR MDA-5, resulting in induction of a strong type I interferon (IFN response. Bacterial flagellin stimulates TLR5 and when delivered intracellularly the cytosolic PRR NLRC4, leading to secretion of proinflammatory cytokines. Both alphaviruses and flagellin have independently been shown to act as adjuvants for antigen-specific antibody responses. Here, we hypothesized that alphavirus and flagellin would act in synergy when combined. We therefore cloned the Salmonella Typhimurium flagellin (FliC gene into an alphavirus replicon and assessed its adjuvant activity on the antibody response against co-administered antigen. In mice immunized with recombinant alphavirus, antibody responses were greatly enhanced compared to soluble FliC or control alphavirus. Both IgG1 and IgG2a/c responses were increased, indicating an enhancement of both Th1 and Th2 type responses. The adjuvant activity of FliC-expressing alphavirus was diminished but not abolished in the absence of TLR5 or type I IFN signaling, suggesting the contribution of several signaling pathways and some synergistic and redundant activity of its components. Thus, we have created a recombinant adjuvant that stimulates multiple signaling pathways of innate immunity resulting in a strong and broad antibody response.

  10. Borreliacidal activity of Borrelia metal transporter A (BmtA binding small molecules by manganese transport inhibition

    Wagh D

    2015-02-01

    Full Text Available Dhananjay Wagh,* Venkata Raveendra Pothineni,* Mohammed Inayathullah, Song Liu, Kwang-Min Kim, Jayakumar Rajadas Biomaterials and Advanced Drug Delivery Laboratory, Stanford Cardiovascular Pharmacology Division, Cardiovascular Institute, Stanford University School of Medicine, Palo Alto, CA, USA *These authors contributed equally to this work  Abstract: Borrelia burgdorferi, the causative agent of Lyme disease, utilizes manganese (Mn for its various metabolic needs. We hypothesized that blocking Mn transporter could be a possible approach to inhibit metabolic activity of this pathogen and eliminate the infection. We used a combination of in silico protein structure prediction together with molecular docking to target the Borrelia metal transporter A (BmtA, a single known Mn transporter in Borrelia and screened libraries of FDA approved compounds that could potentially bind to the predicted BmtA structure with high affinity. Tricyclic antihistamines such as loratadine, desloratadine, and 3-hydroxydesloratadine as well as yohimbine and tadalafil demonstrated a tight binding to the in silico folded BmtA transporter. We, then, tested borreliacidal activity and dose response of the shortlisted compounds from this screen using a series of in vitro assays. Amongst the probed compounds, desloratadine exhibited potent borreliacidal activity in vitro at and above 78 µg/mL (250 µM. Borrelia treated with lethal doses of desloratadine exhibited a significant loss of intracellular Mn specifically and a severe structural damage to the bacterial cell wall. Our results support the possibility of developing a novel, targeted therapy to treat Lyme disease by targeting specific metabolic needs of Borrelia.  Keywords: Lyme disease, BmtA, Borrelia burgdorferi, desloratadine, Bac Titer-Glo assay

  11. Teleconnection between the South Atlantic convergence zone and the southern Indian Ocean: Implications for tropical cyclone activity

    DeBlander, Evan; Shaman, Jeffrey

    2017-01-01

    A link between anomalous austral summer convection over the South Atlantic convergence zone (SACZ) and atmospheric conditions over the South Indian Ocean is investigated. Interannual north-south shifts in the SACZ produce a dipole of anomalous convection and precipitation over South America. The South Atlantic convergence zone index (SACZI) capturing this variability is presented and associated with a midlatitude Rossby wave train that propagates from South America eastward and south of Africa before curving north into the tropical Indian Ocean. This wave train is reproduced using Rossby wave ray tracing and simulations with a linearized barotropic vorticity equation model forced with divergence and convergence consistent with the observed dipole of anomalous convection. The wave train acts to excite anomalies in wind shear and relative humidity over the tropical south Indian Ocean, which in turn impact tropical cyclone (TC) genesis and distribution over this region. We find that changes in the wind shear and relative humidity associated with the SACZI effect changes in TC genesis, which result in a change in observed TC days over portions of the South Indian Ocean.

  12. Linking mantle dynamics, plate tectonics and surface processes in the active plate boundary zones of eastern New Guinea (Invited)

    Baldwin, S.; Moucha, R.; Fitzgerald, P. G.; Hoke, G. D.; Bermudez, M. A.; Webb, L. E.; Braun, J.; Rowley, D. B.; Insel, N.; Abers, G. A.; Wallace, L. M.; Vervoort, J. D.

    2013-12-01

    Eastern New Guinea lies within the rapidly obliquely converging Australian (AUS)- Pacific (PAC) plate boundary zone and is characterized by transient plate boundaries, rapidly rotating microplates and a globally significant geoid high. As the AUS plate moved northward in the Cenozoic, its leading edge has been a zone of subduction and arc accretion. The variety of tectonic settings in this region permits assessment of the complex interplay among mantle dynamics, plate tectonics, and surface processes. Importantly, the timescale of tectonic events (e.g., subduction, (U)HP exhumation, seafloor spreading) are within the valid bounds of mantle convection models. A record of changes in bathymetry and topography are preserved in high standing mountain belts, exhumed extensional gneiss domes and core complexes, uplifted coral terraces, and marine sedimentary basins. Global seismic tomography models indicate accumulation of subducted slabs beneath eastern New Guinea at the bottom of the upper mantle (i.e., 250-300 km). Preliminary global-scale backward advected mantle convection models, driven by density inferred from joint seismic-geodynamic tomography models, exhibit large-scale flow associated with these subducted slab remnants and predict the timing and magnitude (up to 1500 m) of dynamic topography change (both subsidence and uplift) since the Oligocene. In this talk we will explore the effects of large-scale background mantle flow and plate tectonics on the evolution of topography and bathymetry in eastern New Guinea, and discuss possible mechanisms to explain basin subsidence and surface uplift in the region.

  13. The Influence of Vegetation Canopy Structure on Active Layer Thaw Within the Sub-Arctic Discontinuous Permafrost Zone

    Chasmer, L.; Quinton, W.; Hopkinson, C.; Petrone, R.; Whittington, P.

    2009-05-01

    Much of the sub-arctic discontinuous permafrost zone is dominated by a range in peatland ecosystems, each with their own characteristic soil frost dynamics. Soil thaw within the discontinuous permafrost zones of the Canadian sub-arctic is driven by the surface energy balance. The following study examines the influence of canopy structure on frost table (FT) depth and rates of thaw by: 1. relating measurements of FT depth to canopy structure using airborne scanning light detection and ranging (lidar) and hemispherical photographs taken below vegetated canopies; and 2. quantifying the spatial influences of canopy structural characteristics on the radiation balance (direct and diffuse incident radiation) within raised peat plateaus, connected bogs, fens, and isolated bogs. The results of this study indicate that peat plateaus, being characterised by greater vegetation fractional cover, typically have shallower FT depths (r2 = 0.5, p = 0.03) than locations with lower biomass. Further, average ground surface elevation and canopy height are related to rates of FT thaw (r2 = 0.73, p indicates that rates of thaw at the edges of peat plateaus and areas surrounding isolated bogs will be exacerbated by increased incident radiation and less shadowing by the canopy, leading to the conversion of peat plateaus to fens or bogs. This hypothesis is tested by comparing the change in peat plateau area coverage in 2000 and 2008 using classified IKONOS imagery (2000) and airborne lidar (2008).

  14. TGF-β: An Important Mediator of Allergic Disease and a Molecule with Dual Activity in Cancer Development

    Belen Tirado-Rodriguez

    2014-01-01

    Full Text Available The transforming growth factor-β (TGF-β superfamily is a family of structurally related proteins that includes TGF-β, activins/inhibins, and bone morphogenic proteins (BMPs. Members of the TGF-β superfamily regulate cellular functions such as proliferation, apoptosis, differentiation, and migration and thus play key roles in organismal development. TGF-β is involved in several human diseases, including autoimmune disorders and vascular diseases. Activation of the TGF-β receptor induces phosphorylation of serine/threonine residues and triggers phosphorylation of intracellular effectors (Smads. Once activated, Smad proteins translocate to the nucleus and induce transcription of their target genes, regulating various processes and cellular functions. Recently, there has been an attempt to correlate the effect of TGF-β with various pathological entities such as allergic diseases and cancer, yielding a new area of research known as “allergooncology," which investigates the mechanisms by which allergic diseases may influence the progression of certain cancers. This knowledge could generate new therapeutic strategies aimed at correcting the pathologies in which TGF-β is involved. Here, we review recent studies that suggest an important role for TGF-β in both allergic disease and cancer progression.

  15. Activation of VCAM-1 and Its Associated Molecule CD44 Leads to Increased Malignant Potential of Breast Cancer Cells

    Pei-Chen Wang

    2014-02-01

    Full Text Available VCAM-1 (CD106, a transmembrane glycoprotein, was first reported to play an important role in leukocyte adhesion, leukocyte transendothelial migration and cell activation by binding to integrin VLA-1 (α4β1. In the present study, we observed that VCAM-1 expression can be induced in many breast cancer epithelial cells by cytokine stimulation in vitro and its up-regulation directly correlated with advanced clinical breast cancer stage. We found that VCAM-1 over-expression in the NMuMG breast epithelial cells controls the epithelial and mesenchymal transition (EMT program to increase cell motility rates and promote chemoresistance to doxorubicin and cisplatin in vitro. Conversely, in the established MDAMB231 metastatic breast cancer cell line, we confirmed that knockdown of endogenous VCAM-1 expression reduced cell proliferation and inhibited TGFβ1 or IL-6 mediated cell migration, and increased chemosensitivity. Furthermore, we demonstrated that knockdown of endogenous VCAM-1 expression in MDAMB231 cells reduced tumor formation in a SCID xenograft mouse model. Signaling studies showed that VCAM-1 physically associates with CD44 and enhances CD44 and ABCG2 expression. Our findings uncover the possible mechanism of VCAM-1 activation facilitating breast cancer progression, and suggest that targeting VCAM-1 is an attractive strategy for therapeutic intervention.

  16. IGF-I regulates redox status in breast cancer cells by activating the amino acid transport molecule xC-.

    Yang, Yuzhe; Yee, Douglas

    2014-04-15

    Insulin-like growth factors (IGF) stimulate cell growth in part by increasing amino acid uptake. xCT (SLC7A11) encodes the functional subunit of the cell surface transport system xC(-), which mediates cystine uptake, a pivotal step in glutathione synthesis and cellular redox control. In this study, we show that IGF-I regulates cystine uptake and cellular redox status by activating the expression and function of xCT in estrogen receptor-positive (ER(+)) breast cancer cells by a mechanism that relies on the IGF receptor substrate-1 (IRS-1). Breast cancer cell proliferation mediated by IGF-I was suppressed by attenuating xCT expression or blocking xCT activity with the pharmacologic inhibitor sulfasalazine (SASP). Notably, SASP sensitized breast cancer cells to inhibitors of the type I IGF receptor (IGF-IR) in a manner reversed by the reactive oxygen species (ROS) scavenger N-acetyl-L-cysteine. Thus, IGF-I promoted the proliferation of ER(+) breast cancer cells by regulating xC(-) transporter function to protect cancer cells from ROS in an IRS-1-dependent manner. Our findings suggest that inhibiting xC(-) transporter function may synergize with modalities that target the IGF-IR to heighten their therapeutic effects.

  17. Establishing the soundness of administrative spatial units for operationalising the active living potential of residential environments: an exemplar for designing optimal zones

    Riva Mylène

    2008-07-01

    Full Text Available Abstract Background In health and place research, definitions of areas, area characteristics, and health outcomes should ideally be coherent with one another. Yet current approaches for delimiting areas mostly rely on spatial units "of convenience" such as census tracts. These areas may be homogeneous along socioeconomic conditions but heterogeneous along other environmental characteristics. This heterogeneity can lead to biased measurement of environment characteristics and misestimation of area effects on health. The objective of this study was to assess the soundness of census tracts as units of analysis for measuring the active living potential of environments, hypothesised to be associated with walking. Results Starting with data at the smallest census area level available, zones homogeneous along three indicators of active living potential, i.e. population density, land use mix, and accessibility to services were designed. Delimitation of zones ensued from statistical clustering of the smallest areas into seven clusters or "types of environment". Mapping of clusters into a GIS led to the delineation of 898 zones characterised by one of seven types of environment, corresponding to different levels of active living potential. Homogeneity of census tracts along indicators of active living potential varied. A greater proportion (83% of variation in accessibility to services was attributable to differences between census tracts suggesting within-tract homogeneity along this variable. However, census tracts were heterogeneous with respect to population density and land use mix where a greater proportion of the variation was attributable to within-tract differences. About 55% of tracts were characterised by a combination of three or more "types of environment" suggesting substantial within-tract heterogeneity in the active living potential of environments. Conclusion Soundness of census tracts for measuring active living potential may be limited

  18. Disruption of Nrf2 Enhances Upregulation of Nuclear Factor-κB Activity, Proinflammatory Cytokines, and Intercellular Adhesion Molecule-1 in the Brain after Traumatic Brain Injury

    Wei Jin

    2008-01-01

    Full Text Available Inflammatory response plays an important role in the pathogenesis of secondary brain injury after traumatic brain injury (TBI. Nuclear factor erythroid 2-related factor 2 (Nrf2 is a key transcription factor that plays a crucial role in cytoprotection against inflammation. The present study investigated the role of Nrf2 in the cerebral upregulation of NF-κB activity, proinflammatory cytokine, and ICAM-1 after TBI. Wild-type Nrf2 (+/+ and Nrf2 (−/−-deficient mice were subjected to a moderately severe weight-drop impact head injury. Electrophoretic mobility shift assays (EMSAs were performed to analyze the activation of nuclear factor kappa B (NF-κB. Enzyme-linked immunosorbent assays were performed to quantify the production of tumor necrosis factor-α (TNF-α, interleukin-1β (IL-1β, and interleukin-6 (IL-6. Immunohistochemistry staining experiments were performed to detect the expression of intercellular adhesion molecule-1 (ICAM-1. Nrf2 (−/− mice were shown to have more NF-κB activation, inflammatory cytokines TNF-α, IL-1β and IL-6 production, and ICAM-1 expression in brain after TBI compared with their wild-type Nrf2 (+/+ counterparts. The results suggest that Nrf2 plays an important protective role in limiting the cerebral upregulation of NF-κB activity, proinflammatory cytokine, and ICAM-1 after TBI.

  19. Common catabolic enzyme patterns in a microplankton community of the Humboldt Current System off northern and central-south Chile: Malate dehydrogenase activity as an index of water-column metabolism in an oxygen minimum zone

    González, R. R.; Quiñones, R. A.

    2009-07-01

    An extensive subsurface oxygen minimum zone off northern and central-south Chile, associated with the Peru-Chile undercurrent, has important effects on the metabolism of the organisms inhabiting therein. Planktonic species deal with the hypoxic and anoxic environments by relying on biochemical as well as physiological processes related to their anaerobic metabolisms. Here we characterize, for the first time, the potential enzymatic activities involved in the aerobic and anaerobic energy production pathways of microplanktonic organisms (catabolic pathways in the oxygen minimum zone. Malate dehydrogenase had the highest oxidizing activity of nicotinamide adenine dinucleotide (reduced form) in the batch of catabolic enzymatic activities assayed, including potential pyruvate oxidoreductases activity, the electron transport system, and dissimilatory nitrate reductase. Malate dehydrogenase correlated significantly with almost all the enzymes analyzed within and above the oxygen minimum zone, and also with the oxygen concentration and microplankton biomass in the water column of the Humboldt Current System, especially in the oxygen minimum zone off Iquique. These results suggest a possible specific pattern for the catabolic activity of the microplanktonic realm associated with the oxygen minimum zone spread along the Humboldt Current System off Chile. We hypothesize that malate dehydrogenase activity could be an appropriate indicator of microplankton catabolism in the oxygen minimum zone and adjacent areas.

  20. Fully relativistic complete active space self-consistent field for large molecules: Quasi-second-order minimax optimization

    Bates, Jefferson E.; Shiozaki, Toru [Department of Chemistry, Northwestern University, 2145 Sheridan Rd., Evanston, Illinois 60208 (United States)

    2015-01-28

    We develop an efficient algorithm for four-component complete active space self-consistent field (CASSCF) methods on the basis of the Dirac equation that takes into account spin–orbit and other relativistic effects self-consistently. Orbitals are optimized using a trust-region quasi-Newton method with Hessian updates so that energies are minimized with respect to rotations among electronic orbitals and maximized with respect to rotations between electronic and positronic orbitals. Utilizing density fitting and parallel computation, we demonstrate that Dirac–Coulomb CASSCF calculations can be routinely performed on systems with 100 atoms and a few heavy-elements. The convergence behavior and wall times for octachloridodirhenate(III) and a tungsten methylidene complex are presented. In addition, the excitation energies of octachloridodirhenate(III) are reported using a state-averaged variant.

  1. Fully relativistic complete active space self-consistent field for large molecules: Quasi-second-order minimax optimization

    Bates, Jefferson E.; Shiozaki, Toru

    2015-01-01

    We develop an efficient algorithm for four-component complete active space self-consistent field (CASSCF) methods on the basis of the Dirac equation that takes into account spin-orbit and other relativistic effects self-consistently. Orbitals are optimized using a trust-region quasi-Newton method with Hessian updates so that energies are minimized with respect to rotations among electronic orbitals and maximized with respect to rotations between electronic and positronic orbitals. Utilizing density fitting and parallel computation, we demonstrate that Dirac-Coulomb CASSCF calculations can be routinely performed on systems with 100 atoms and a few heavy-elements. The convergence behavior and wall times for octachloridodirhenate(III) and a tungsten methylidene complex are presented. In addition, the excitation energies of octachloridodirhenate(III) are reported using a state-averaged variant.

  2. Satellites of Xe transitions induced by infrared active vibrational modes of CF4 and C2F6 molecules.

    Alekseev, Vadim A; Schwentner, Nikolaus

    2011-07-28

    Absorption and luminescence excitation spectra of Xe/CF(4) mixtures were studied in the vacuum UV region at high resolution using tunable synchrotron radiation. Pressure-broadened resonance bands and bands associated with dipole-forbidden states of the Xe atom due to collision-induced breakdown of the optical selection rules are reported. The spectra display in addition numerous satellite bands corresponding to transitions to vibrationally excited states of a Xe-CF(4) collisional complex. These satellites are located at energies of Xe atom transition increased by one quantum energy in the IR active v(3) vibrational mode of CF(4) (v(3) = 1281 cm(-1)). Satellites of both resonance and dipole-forbidden transitions were observed. Satellites of low lying resonance states are spectrally broad bands closely resembling in shape their parent pressure-broadened resonance bands. In contrast, satellites of dipole-forbidden states and of high lying resonance states are spectrally narrow bands (FWHM ∼10 cm(-1)). The satellites of dipole-forbidden states are orders of magnitude stronger than transitions to their parent states due to collision-induced breakdown of the optical selection rules. These satellites are attributed to a coupling of dipole-forbidden and resonance states induced by the electric field of the transient CF(4) (v(3) = 0 ↔ v(3) = 1) dipole. Similar satellites are present in spectra of Xe/C(2)F(6) mixtures where these bands are induced by the IR active v(10) mode of C(2)F(6). Transitions to vibrationally excited states of Xe-CF(4)(C(2)F(6)) collision pairs were also observed in two-photon LIF spectra.

  3. Proteins Are the Body's Worker Molecules

    ... PDF Chapter 1: Proteins are the Body's Worker Molecules You've probably heard that proteins are important ... are much more than that. Proteins are worker molecules that are necessary for virtually every activity in ...

  4. Quantum Chemical Study of the Solvent Effect on the Anticancer Active Molecule of Iproplatin: Structural, Electronic, and Spectroscopic Properties (IR, 1H NMR, UV)

    Sadeghi, N.; Ghiasi, R.; Fazaeli, R.; Jamehbozorgi, S.

    2017-01-01

    The structural, electronic, and spectroscopic properties of the anticancer active molecule of iproplatin were investigated in the gas and liquid phases. Based on the polarizable continuum model (PCM), the solvent effect on the structural parameters, frontier orbitals, and spectroscopic parameters of the complex was investigated. The results indicate that the polarity of solvents plays a significant role in the structure and pro perties of the complex. 1H and 13C NMR chemical shifts were calculated using the Gauge-invariant atomic orbital (GIAO) method. Pt-Cl and Pt-OH bonds were investigated through a vibrational analysis. Moreover, time dependent density functional theory (TD-DFT) was used to calculate the energy, oscillatory strength, and wavelength absorption maximum (λmax) of electronic transitions and its nature within the complex.

  5. A Selective Assay to Detect Chitin and Biologically Active Nano-Machineries for Chitin-Biosynthesis with Their Intrinsic Chitin-Synthase Molecules

    Hildgund Schrempf

    2010-09-01

    Full Text Available A new assay system for chitin has been developed. It comprises the chitin-binding protein ChbB in fusion with a His-tag as well as with a Strep-tag, the latter of which was chemically coupled to horseradish peroxidase. With the resulting complex, minimal quantities of chitin are photometrically detectable. In addition, the assay allows rapid scoring of the activity of chitin-synthases. As a result, a refined procedure for the rapid purification of yeast chitosomes (nano-machineries for chitin biosynthesis has been established. Immuno-electronmicroscopical studies of purified chitosomes, gained from a yeast strain carrying a chitin-synthase gene fused to that for GFP (green-fluorescence protein, has led to the in situ localization of chitin-synthase-GFP molecules within chitosomes.

  6. Identification of amino acid residues involved in the interaction between measles virus Haemagglutin (MVH) and its human cell receptor (signaling lymphocyte activation molecule, SLAM).

    Xu, Qin; Zhang, Peng; Hu, Chunling; Liu, Xin; Qi, Yipeng; Liu, Yingle

    2006-07-31

    Signaling lymphocyte activation molecule (SLAM; also known as CD150) is a newly identified cellular receptor for measles virus (MV). The interaction between MV Haemagglutin (MVH) and SLAM is an initial step for MV entry. We have identified several novel SLAM binding sites at residues S429, T436 and H437 of MVH protein and MVH mutants in these residues dramatically decrease the ability to interaction with the cell surface SLAM and fail to coprecipitation with SLAM in vivo as well as malfunction in syncytium formation. At the same time, K58, S59 and H61 of SLAM was also identified to be critical for MVH and SLAM binding. Further, these residues may be useful targets for the development of measles therapy.

  7. Activation energy of fractional vortices and spectroscopy of a vortex molecule in long Josephson junction; Aktivierungsenergie fraktionaler Flusswirbel und Spektroskopie an Vortex-Molekuelen in langen Josephsonkontakten

    Buckenmaier, Kai

    2010-06-09

    This thesis is divided into two parts, the measurement of the activation energy of a fractional vortex and the spectroscopy of a vortex-molecule. Fractional vortices can be studied in long 0-{kappa} Josephson junctions, where a jump of the Josephson phase is created artificially with a pair of tiny current injectors. To compensate for this phase discontinuity, a {rho} vortex is formed. Here, {rho} describes the vortex's so called topological charge. The {rho} vortices are pinned at the discontinuity and they carry the fraction ({rho}/2).{phi}{sub 0} of magnetic flux, with the magnetic flux quantum {phi}{sub 0} 2.07.10{sup -15}. Two stable vortex configurations are possible, a direct Vortex and a complementary one. {rho} depends on the injector current. When the bias current of the junction exceeds a characteristic threshold, which dependents on {rho}, the Lorentz force is bigger than the pinning force of the vortex and a fluxon is pulled away. In this case a complementary ({rho}-2{pi}) vortex is left behind. This switching of the {rho} vortex and the resulting emission of a fluxon can be described as a Kramers like escape of a particle out of a tilted washboard potential. The washboard potential is tilted to the point where the barrier is small enough, so that the particle can escape via thermal or quantum fluctuations. In the case of thermal fluctuations the barrier height is called activation energy. The activation energy can be determined by measuring the junction's switching current statistics. In this thesis, the activation energy, necessary for the vortex escape, was measured as a function of {rho} and a homogenous external magnetic field perpendicular to the junction. The main focus was the investigation of 0-{pi} junctions. The temperature dependence of the activation energy was investigated, too. It turns out, that the transition-state-theory is convenient to describe the switching probability of the standard Nb-AlO{sub x}-Nb junctions at 4.2 K

  8. Identification and Small Molecule Inhibition of an Activating Transcription Factor 4 (ATF4)-dependent Pathway to Age-related Skeletal Muscle Weakness and Atrophy*

    Ebert, Scott M.; Dyle, Michael C.; Bullard, Steven A.; Dierdorff, Jason M.; Murry, Daryl J.; Fox, Daniel K.; Bongers, Kale S.; Lira, Vitor A.; Meyerholz, David K.; Talley, John J.; Adams, Christopher M.

    2015-01-01

    Aging reduces skeletal muscle mass and strength, but the underlying molecular mechanisms remain elusive. Here, we used mouse models to investigate molecular mechanisms of age-related skeletal muscle weakness and atrophy as well as new potential interventions for these conditions. We identified two small molecules that significantly reduce age-related deficits in skeletal muscle strength, quality, and mass: ursolic acid (a pentacyclic triterpenoid found in apples) and tomatidine (a steroidal alkaloid derived from green tomatoes). Because small molecule inhibitors can sometimes provide mechanistic insight into disease processes, we used ursolic acid and tomatidine to investigate the pathogenesis of age-related muscle weakness and atrophy. We found that ursolic acid and tomatidine generate hundreds of small positive and negative changes in mRNA levels in aged skeletal muscle, and the mRNA expression signatures of the two compounds are remarkably similar. Interestingly, a subset of the mRNAs repressed by ursolic acid and tomatidine in aged muscle are positively regulated by activating transcription factor 4 (ATF4). Based on this finding, we investigated ATF4 as a potential mediator of age-related muscle weakness and atrophy. We found that a targeted reduction in skeletal muscle ATF4 expression reduces age-related deficits in skeletal muscle strength, quality, and mass, similar to ursolic acid and tomatidine. These results elucidate ATF4 as a critical mediator of age-related muscle weakness and atrophy. In addition, these results identify ursolic acid and tomatidine as potential agents and/or lead compounds for reducing ATF4 activity, weakness, and atrophy in aged skeletal muscle. PMID:26338703

  9. Exosomes secreted by human placenta carry functional Fas ligand and TRAIL molecules and convey apoptosis in activated immune cells, suggesting exosome-mediated immune privilege of the fetus.

    Stenqvist, Ann-Christin; Nagaeva, Olga; Baranov, Vladimir; Mincheva-Nilsson, Lucia

    2013-12-01

    Apoptosis is crucially important in mediating immune privilege of the fetus during pregnancy. We investigated the expression and in vitro apoptotic activity of two physiologically relevant death messengers, the TNF family members Fas ligand (FasL) and TRAIL in human early and term placentas. Both molecules were intracellularly expressed, confined to the late endosomal compartment of the syncytiotrophoblast, and tightly associated to the generation and secretion of placental exosomes. Using immunoelectron microscopy, we show that FasL and TRAIL are expressed on the limiting membrane of multivesicular bodies where, by membrane invagination, intraluminal microvesicles carrying membranal bioactive FasL and TRAIL are formed and released in the extracellular space as exosomes. Analyzing exosomes secreted from placental explant cultures, to our knowledge, we demonstrate for the first time that FasL and TRAIL are clustered on the exosomal membrane as oligomerized aggregates ready to form death-inducing signaling complex. Consistently, placental FasL- and TRAIL-carrying exosomes triggered apoptosis in Jurkat T cells and activated PBMC in a dose-dependent manner. Limiting the expression of functional FasL and TRAIL to exosomes comprise a dual benefit: 1) storage of exosomal FasL and TRAIL in multivesicular bodies is protected from proteolytic cleavage and 2) upon secretion, delivery of preformed membranal death molecules by exosomes rapidly triggers apoptosis. Our results suggest that bioactive FasL- and TRAIL-carrying exosomes, able to convey apoptosis, are secreted by the placenta and tie up the immunomodulatory and protective role of human placenta to its exosome-secreting ability.

  10. Dinamic changes of pro-inflammatory cytokines, adhesion molecules and lymphocytes activation markers as early indicators of diseases severity in patients with Dengue

    Silvana Vielma

    2014-09-01

    Full Text Available Several immunopathogenic mechanisms have been proposed to explain the massive increase of vascular permeability observed in the severe forms of infection by Dengue Virus (DENV. Our aim was to determine the kinetic changes of inflammatory mediators (IL-8, TNF- α, soluble early lymphocyte activation markers (sIL-2R, sTNF-Rp75 and soluble fractions of cell adhesion molecules (sICAM-1 and sVCAM-1 as indicators for early recognition of disease severity in patients with laboratory-confirmed dengue. Twenty patients classified as Dengue±Warning Signs (D±WS and thirty patients with Severe Dengue (SD were included in the study. Serums of apparently healthy individuals were included as controls. Compared with normal subjects, D±WS cases did not show significant differences in the levels of IL-8 or TNF-α during the acute nor in the critical stages of the disease; however, in D±WS cases levels of sICAM-1 and sVCAM-1 were higher than controls during both phases; in contrast, significant increase of sTNF-p75 and sIL2R levels were observed during the critical phase of the disease. Compared with both dengue patients and controls, patients with SD showed significant rise in the levels of IL-8 and TNF-α during the critical phase of the disease and a significant increase in adhesion molecules were detected in both phases, but the highest levels of sVCAM-1 and sIL-2R were observed only during the acute stage of the disease. In conclusion, sIL-2R and sVCAM-1, as early markers of lymphocyte and endothelial activation, would serves as indicators of severity during the acute phase of dengue infection.

  11. Hyperthermostable binding molecules on phage: Assay components for point-of-care diagnostics for active tuberculosis infection.

    Zhao, Ning; Spencer, John; Schmitt, Margaret A; Fisk, John D

    2017-03-15

    Tuberculosis is the leading cause of death from infectious disease worldwide. The low sensitivity, extended processing time, and high expense of current diagnostics are major challenges to the detection and treatment of tuberculosis. Mycobacterium tuberculosis ornithine transcarbamylase (Mtb OTC, Rv1656) has been identified in the urine of patients with active TB infection and is a promising target for point-of-care diagnostics. Specific binding proteins with low nanomolar affinities for Mtb OTC were selected from a phage display library built upon a hyperthermostable Sso7d scaffold. Phage particles displaying Sso7d variants were utilized to generate a sandwich ELISA-based assay for Mtb OTC. The assay response is linear between 2 ng/mL and 125 ng/mL recombinant Mtb OTC and has a limit of detection of 400 pg/mL recombinant Mtb OTC. The assay employing a phage-based detection reagent is comparable to commercially-available antibody-based biosensors. Importantly, the assay maintains functionality at both neutral and basic pH in presence of salt and urea over the range of concentrations typical for human urine. Phage-based diagnostic systems may feature improved physical stability and cost of production relative to traditional antibody-based reagents, without sacrificing specificity and sensitivity.

  12. An Antifungal Combination Matrix Identifies a Rich Pool of Adjuvant Molecules that Enhance Drug Activity against Diverse Fungal Pathogens

    Nicole Robbins

    2015-11-01

    Full Text Available There is an urgent need to identify new treatments for fungal infections. By combining sub-lethal concentrations of the known antifungals fluconazole, caspofungin, amphotericin B, terbinafine, benomyl, and cyprodinil with ∼3,600 compounds in diverse fungal species, we generated a deep reservoir of chemical-chemical interactions termed the Antifungal Combinations Matrix (ACM. Follow-up susceptibility testing against a fluconazole-resistant isolate of C. albicans unveiled ACM combinations capable of potentiating fluconazole in this clinical strain. We used chemical genetics to elucidate the mode of action of the antimycobacterial drug clofazimine, a compound with unreported antifungal activity that synergized with several antifungals. Clofazimine induces a cell membrane stress for which the Pkc1 signaling pathway is required for tolerance. Additional tests against additional fungal pathogens, including Aspergillus fumigatus, highlighted that clofazimine exhibits efficacy as a combination agent against multiple fungi. Thus, the ACM is a rich reservoir of chemical combinations with therapeutic potential against diverse fungal pathogens.

  13. Effect of androgen withdrawal on activation of ERKs and expression of cell cycle regulation molecules in human prostate carcinoma cells

    YE Ding-wei; LI Hui; TSENG Jane; CHAUVIN Priscilla; QIAN Song-xi; ZHENG Jia-fu; SUN Ying-hao; MA Yong-jiang

    2002-01-01

    Objective: To explore the possible mechanisms of growth regression of human androgen dependentprostate carcinoma cells caused by androgen withdrawal. Methods: After 24 h of treatment with 1×10-9mol/L dihydrotestosterone (DHT), the expression of phosphorylated ERK proteins and cell cycle regulationmolecules including CDK2, CDK4, CDK6 and P27kip1 in human androgen dependent prostate carcinoma cellline LNCaP was measured by Western blot analysis 0 h, 8 h and 24 h of after androgen withdrawal. Humanandrogen independent prostate carcinoma cell line PC-3 was also examined as control. Results: Down-regula-tion of phosphorylated ERK, CDK2, CDK4 and CDK6 and up-regulation of P27kip1 were found initially inLNCaP cell line 8 h after androgen withdrawal. The levels of phosphorylated ERK and CDKs decreased con-tinuously and reached the lowest after 24 h, while continuous elevation of P27kip1 was detected thereafter to 24h. No expression change of phosphorylated ERK, CDKs and P27kip1 were detected in PC-3 cell line. Conclu-sion: The androgen withdrawal can cause ERKs activation decrease and cell cycle regulation moleculeschanges, which may be one of the mechanisms for inhibited growth of androgen dependent prostate carcinomaafter androgen ablation by either operative or medicine methods.

  14. The role of sexual vs. asexual recruitment of Artemisia wudanica in transition zone habitats between inter-dune lowlands and active dunes in Inner Mongolia, China

    Wang, Yongcui; Alberto, Busso Carlos; Jiang, Deming; Ala, Musa; Li, Xuehua; Zhou, Quanlai; Lin, Jixiang; Ren, Guohua; Jia, Lian

    2016-04-01

    Artemisia wudanica is an endemic, perennial, pioneering psammophyte species in the sand dune ecosystems of western Horqin Sand Land in northern China. However, no studies have addressed how sexual and asexual reproduction modes of A. wudanica perform at the transitional zones between active dune inter-dune lowlands and active dunes. In early spring, quadrats were randomly set up in the study area to monitor surviving seedling and/or ramet density and frequency coming from sexual/asexual reproduction of A. wudanica. Iron sticks were inserted near each quadrat to determine wind erosion intensity (WE). Additionally, soil samples were collected nearby each quadrat to test for soil moisture (SM), organic matter (OM) and pH. Surviving seedlings of A. wudanica showed an inverse response in comparison with ramets to SM, OM and WE. Soil moisture showed the most positive effect, and WE the negative effect, on surviving, sexual reproduction seedlings. Contrarily, WE had the most positive effect, and SM the negative effect, on asexual reproduction ramets. This suggests that increases in SM and decreases in WE should benefit recruitment of A. wudanica seedlings. On the contrary, ramets coming from asexual reproduction showed a different response to environmental factors in transition zone habitats. While SM was not a key constraint for the survival of seedlings, they showed a better, positive response to wind erosion environments. Overall, various study environmental parameters could be improved to foster A. wudanica invasion and settlement in the plant community through different reproductive modes, thereby promoting vegetation restoration and rehabilitation.

  15. Geological and hydrogeological conditions of the Aigion seismic active fault zone (Deep Geodynamic Laboratory Corinth) based on borehole data and hydraulic tests

    Rettenmaier, D.; Giurgea, V.; Pizzino, L.; Unkel, I.; Hoetzl, H.; Foerster, A.; Quattrocchi, F.; Nikas, K.

    2003-04-01

    The Gulf of Corinth and the northern part of the Peloponnesus/Greece, an area of asymmetric graben structure, step faults and tilted blocks, is one of the most active seismic zones in the world. Six major faults are known to be most responsible for the historic and present seismic activities in the area of Aigion. Our study focuses preliminarily on the area around the Aigion fault, whose trace runs E-W through the harbour of Aigion. Investigations of the stratigraphic sequence, tectonic structure and hydrogeologic conditions of the southern Corinth graben shoulder and first drilling activities there, have started in summer 2001. From July until September 2002 the International Continental Deep Drilling Project (ICDP) and the EU Project DGLab-Gulf of Corinth drilled the AIG10 borehole in the harbour of Aigion to a total depth of 1001 m. Our investigations in this ICDP/EU framework are aimed at studying the thermal-hydraulic conditions on the southern graben shoulder. Here we report the first results on sampling and hydraulic testing. The deep AIG10 borehole has successfully cored in approx. 760 m depth the fault plane, which separates fractured radiolarite in the hanging wall from highly fractured and karstified platy, micritic limestone (Olonos-Pindos Unit) in the footwall. A complete lithologic section is now available through the monitoring of cuttings and cores, which built a major cornerstone for defining an integrated regional tectonic and geologic model. Several pumping tests and hydrochemical investigations made in the region of Aigion and especially in the AIG10 borehole deliver together with geophysical borehole logging the database for a thermo-hydraulic heat flow model. The pumping test AIG10C in the conglomerates of the graben sediments show a hydraulic conductivity of about 2 x 10E-5 m/s - 3 x 10E-4 m/s at a depth of approximately 211 m. The result was a residual drawdown, which indicates a closed hydraulic system between the semi-permeable Aigion

  16. Biology-oriented synthesis of a natural-product inspired oxepane collection yields a small-molecule activator of the Wnt-pathway.

    Basu, Sudipta; Ellinger, Bernhard; Rizzo, Stefano; Deraeve, Céline; Schürmann, Markus; Preut, Hans; Arndt, Hans-Dieter; Waldmann, Herbert

    2011-04-26

    In Biology Oriented Synthesis the scaffolds of biologically relevant compound classes inspire the synthesis of focused compound collections enriched in bioactivity. This criterion is met by the structurally complex scaffolds of natural products (NPs) selected in evolution. The synthesis of NP-inspired compound collections approaching the complexity of NPs calls for the development of efficient synthetic methods. We have developed a one pot 4-7 step synthesis of mono-, bi-, and tricyclic oxepanes that resemble the core scaffolds of numerous NPs with diverse bioactivities. This sequence entails a ring-closing ene-yne metathesis reaction as key step and makes productive use of polymer-immobilized scavenger reagents. Biological profiling of a corresponding focused compound collection in a reporter gene assay monitoring for Wnt-signaling modulation revealed active Wntepanes. This unique class of small-molecule activators of the Wnt pathway modulates the van-Gogh-like receptor proteins (Vangl), which were previously identified in noncanonical Wnt signaling, and acts in synergy with the canonical activator protein (Wnt-3a).

  17. Modulation of glucokinase by glucose, small-molecule activator and glucokinase regulatory protein: steady-state kinetic and cell-based analysis.

    Bourbonais, Francis J; Chen, Jing; Huang, Cong; Zhang, Yanwei; Pfefferkorn, Jeffrey A; Landro, James A

    2012-02-01

    GK (glucokinase) is an enzyme central to glucose metabolism that displays positive co-operativity to substrate glucose. Small-molecule GKAs (GK activators) modulate GK catalytic activity and glucose affinity and are currently being pursued as a treatment for Type 2 diabetes. GK progress curves monitoring product formation are linear up to 1 mM glucose, but biphasic at 5 mM, with the transition from the lower initial velocity to the higher steady-state velocity being described by the rate constant kact. In the presence of a liver-specific GKA (compound A), progress curves at 1 mM glucose are similar to those at 5 mM, reflecting activation of GK by compound A. We show that GKRP (GK regulatory protein) is a slow tight-binding inhibitor of GK. Analysis of progress curves indicate that this inhibition is time dependent, with apparent initial and final Ki values being 113 and 12.8 nM respectively. When GK is pre-incubated with glucose and compound A, the inhibition observed by GKRP is time dependent, but independent of GKRP concentration, reflecting the GKA-controlled transition between closed and open GK conformations. These data are supported by cell-based imaging data from primary rat hepatocytes. This work characterizes the modulation of GK by a novel GKA that may enable the design of new and improved GKAs.

  18. Lipopolysaccharide Binding Protein, Soluble-Intercellular Adhesion Molecule-1, Procalcitonin, and Protein C Activity and Clinical Outcome in Systemic Inflammatory Response Syndrome (SIRS or Sepsis Patients

    Dewi Muliaty

    2009-04-01

    Full Text Available BACKGROUND: Biochemical markers may be used in diagnosis, prognostic and monitoring treatment and therapy for sepsis patients. In this study we used Lipopolysacharide Binding Protein (LBP, serum-Intercellular Adhesion Molecule-1 (ICAM-1, Procalcitonin (PCT and protein C activity. LBP is related to lipopolysachharide or gram-negative bacterial endotoxin which bound to LBP and induced inflammatory response. ICAM-1 is associated with endothelial dysfunction in response to systemic inflammatory and septic condition. PCT increased in bacterial infection and in severe systemic inflammatory. Role of Protein C is protecting the intravascular system to systemic inflammation, sepsis and the concomitant intravascular coagulopathy. The aim of this study was to examine the associations between levels of serum LBP, sICAM-1, PCT, and protein C activity with the clinical outcome of SIRS or sepsis patients. METHODS: We included 19 post surgery patients with SIRS criteria from intensive care unit (ICU and evaluated the level of LBP serum with Chemiliuminescent Enzyme Immunoassay (Diagnostic Product Co., ICAM-1 with ELISA (R&D System, PCT with immunochromatography (BRAHMS, protein C activity with chromogenic method (Dade Behring. We performed the samples serially at the first admission of patients and after 72 hours. Data were analysed by non-parametric with Wilcoxon test and Mann-Whitney test. Correlation study between biomarkers calculated by Kendall’s tau and Spearman’s rho. RESULTS: Of 19 patients, 9 (47,4% died and 10 (52,6% surviving. The level of LBP serum decreased after 72 hours in surviving-sepsis patients, and increased in nonsurviving sepsis patients with significant different levels at 72 hours examination (p0.05. In all patients were found high level of PCT serum since the first admission examination, decreasing levels were occurred significantly in surviving patients after 72 hours (p0.05 both in surviving and non-surviving patients. CONCLUSIONS

  19. Efficiency of Pumping of the Active Medium of Metal Vapor Lasers: Gas-Discharge Tubes with Electrodes in the Hot Zone of the Discharge Channel