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Sample records for dag fjeld iwan

  1. Simulation of dynamics of beam structures with bolted joints using adjusted Iwan beam elements

    NASA Astrophysics Data System (ADS)

    Song, Y.; Hartwigsen, C. J.; McFarland, D. M.; Vakakis, A. F.; Bergman, L. A.

    2004-05-01

    Mechanical joints often affect structural response, causing localized non-linear stiffness and damping changes. As many structures are assemblies, incorporating the effects of joints is necessary to produce predictive finite element models. In this paper, we present an adjusted Iwan beam element (AIBE) for dynamic response analysis of beam structures containing joints. The adjusted Iwan model consists of a combination of springs and frictional sliders that exhibits non-linear behavior due to the stick-slip characteristic of the latter. The beam element developed is two-dimensional and consists of two adjusted Iwan models and maintains the usual complement of degrees of freedom: transverse displacement and rotation at each of the two nodes. The resulting element includes six parameters, which must be determined. To circumvent the difficulty arising from the non-linear nature of the inverse problem, a multi-layer feed-forward neural network (MLFF) is employed to extract joint parameters from measured structural acceleration responses. A parameter identification procedure is implemented on a beam structure with a bolted joint. In this procedure, acceleration responses at one location on the beam structure due to one known impulsive forcing function are simulated for sets of combinations of varying joint parameters. A MLFF is developed and trained using the patterns of envelope data corresponding to these acceleration histories. The joint parameters are identified through the trained MLFF applied to the measured acceleration response. Then, using the identified joint parameters, acceleration responses of the jointed beam due to a different impulsive forcing function are predicted. The validity of the identified joint parameters is assessed by comparing simulated acceleration responses with experimental measurements. The capability of the AIBE to capture the effects of bolted joints on the dynamic responses of beam structures, and the efficacy of the MLFF parameter

  2. Application of viscous and Iwan modal damping models to experimental measurements from bolted structures

    DOE PAGESBeta

    Deaner, Brandon J.; Allen, Matthew S.; Starr, Michael James; Segalman, Daniel J.; Sumali, Hartono

    2015-01-20

    Measurements are presented from a two-beam structure with several bolted interfaces in order to characterize the nonlinear damping introduced by the joints. The measurements (all at force levels below macroslip) reveal that each underlying mode of the structure is well approximated by a single degree-of-freedom (SDOF) system with a nonlinear mechanical joint. At low enough force levels, the measurements show dissipation that scales as the second power of the applied force, agreeing with theory for a linear viscously damped system. This is attributed to linear viscous behavior of the material and/or damping provided by the support structure. At larger forcemore » levels, the damping is observed to behave nonlinearly, suggesting that damping from the mechanical joints is dominant. A model is presented that captures these effects, consisting of a spring and viscous damping element in parallel with a four-parameter Iwan model. As a result, the parameters of this model are identified for each mode of the structure and comparisons suggest that the model captures the stiffness and damping accurately over a range of forcing levels.« less

  3. Application of viscous and Iwan modal damping models to experimental measurements from bolted structures

    SciTech Connect

    Deaner, Brandon J.; Allen, Matthew S.; Starr, Michael James; Segalman, Daniel J.; Sumali, Hartono

    2015-01-20

    Measurements are presented from a two-beam structure with several bolted interfaces in order to characterize the nonlinear damping introduced by the joints. The measurements (all at force levels below macroslip) reveal that each underlying mode of the structure is well approximated by a single degree-of-freedom (SDOF) system with a nonlinear mechanical joint. At low enough force levels, the measurements show dissipation that scales as the second power of the applied force, agreeing with theory for a linear viscously damped system. This is attributed to linear viscous behavior of the material and/or damping provided by the support structure. At larger force levels, the damping is observed to behave nonlinearly, suggesting that damping from the mechanical joints is dominant. A model is presented that captures these effects, consisting of a spring and viscous damping element in parallel with a four-parameter Iwan model. As a result, the parameters of this model are identified for each mode of the structure and comparisons suggest that the model captures the stiffness and damping accurately over a range of forcing levels.

  4. FIRE near-infrared spectroscopic classifications of SN 2016dag

    NASA Astrophysics Data System (ADS)

    Morrell, N.; Phillips, M. M.; Contreras, C.; Hsia, E. Y.

    2016-07-01

    We report the spectroscopic classification of SN 2016dag, discovered by the Backyard Observatory Supernova Search (BOSS), using a near-infrared spectrum (range 800-2500 nm) obtained on Jul 14.95 UT with the FoldedPort Infrared Echellette (FIRE) spectrograph on the 6.5-m Magellan Baade Telescope at Las Campanas Observatory.

  5. Turkey's next big science project: DAG the 4 meter telescope

    NASA Astrophysics Data System (ADS)

    Keskin, O.; Yesilyaprak, C.; Yerli, S. K.; Zago, L.; Jolissaint, L.

    2014-07-01

    The DAG (Turkish for Eastern Anatolia Observatory) 4-m telescope project has been formally launched in 2012, being fully funded by the Government of Turkey. This new observatory is to be located on a 3170 m altitude ridge near the town of Erzurum in Eastern Anatolia. First light is scheduled for late 2017. The DAG team's baseline design of the telescope consists of a Ritchey-Chretien type with alt-az mount, a focal length of 56 m and a field of view up to 30 arcmin. Multiple instruments will be located at the Nasmyth foci. The optical specifications of the telescope are set by DAG team for diffraction limited performance with active and adaptive optics. Modern mirror control technologies will allow defining in a most cost effective way the figuring requirements of the optical surfaces: the low order figuring errors of the combined optical train constituted of M1-M2-M3 are defined in terms of Zernike coefficients and referred to the M1 surface area. The high order figuring errors are defined using the phase structure functions. Daytime chilling of the closed enclosure volume and natural ventilation through suitable openings during observations will be used to ensure optimal mirror and dome seeing. A design of a ground layer adaptive optics (GLAO) subsystem is developed concurrently with the telescope. In this paper, main design aspects, the optical design and expected performance analysis of the telescope will be presented.

  6. DAG-TM Concept Element 11 CNS Performance Assessment: ADS-B Performance in the TRACON

    NASA Technical Reports Server (NTRS)

    Raghavan, Rajesh S.

    2004-01-01

    Distributed Air/Ground (DAG) Traffic Management (TM) is an integrated operational concept in which flight deck crews, air traffic service providers and aeronautical operational control personnel use distributed decision-making to enable user preferences and increase system capacity, while meeting air traffic management (ATM) safety requirements. It is a possible operational mode under the Free Flight concept outlined by the RTCA Task Force 3. The goal of DAG-TM is to enhance user flexibility/efficiency and increase system capacity, without adversely affecting system safety or restricting user accessibility to the National Airspace System (NAS). DAG-TM will be accomplished with a human-centered operational paradigm enabled by procedural and technological innovations. These innovations include automation aids, information sharing and Communication, Navigation, and Surveillance (CNS) / ATM technologies. The DAG-TM concept is intended to eliminate static restrictions to the maximum extent possible. In this paradigm, users may plan and operate according to their preferences - as the rule rather than the exception - with deviations occurring only as necessary. The DAG-TM concept elements aim to mitigate the extent and impact of dynamic NAS constraints, while maximizing the flexibility of airspace operations

  7. Human CalDAG-GEFI gene (RASGRP2) mutation affects platelet function and causes severe bleeding

    PubMed Central

    Canault, Matthias; Ghalloussi, Dorsaf; Grosdidier, Charlotte; Guinier, Marie; Perret, Claire; Chelghoum, Nadjim; Germain, Marine; Raslova, Hana; Peiretti, Franck; Morange, Pierre E.; Saut, Noemie; Pillois, Xavier; Nurden, Alan T.; Cambien, François; Pierres, Anne; van den Berg, Timo K.; Kuijpers, Taco W.; Tregouet, David-Alexandre

    2014-01-01

    The nature of an inherited platelet disorder was investigated in three siblings affected by severe bleeding. Using whole-exome sequencing, we identified the culprit mutation (cG742T) in the RAS guanyl-releasing protein-2 (RASGRP2) gene coding for calcium- and DAG-regulated guanine exchange factor-1 (CalDAG-GEFI). Platelets from individuals carrying the mutation present a reduced ability to activate Rap1 and to perform proper αIIbβ3 integrin inside-out signaling. Expression of CalDAG-GEFI mutant in HEK293T cells abolished Rap1 activation upon stimulation. Nevertheless, the PKC- and ADP-dependent pathways allow residual platelet activation in the absence of functional CalDAG-GEFI. The mutation impairs the platelet’s ability to form thrombi under flow and spread normally as a consequence of reduced Rac1 GTP-binding. Functional deficiencies were confined to platelets and megakaryocytes with no leukocyte alteration. This contrasts with the phenotype seen in type III leukocyte adhesion deficiency caused by the absence of kindlin-3. Heterozygous did not suffer from bleeding and have normal platelet aggregation; however, their platelets mimicked homozygous ones by failing to undergo normal adhesion under flow and spreading. Rescue experiments on cultured patient megakaryocytes corrected the functional deficiency after transfection with wild-type RASGRP2. Remarkably, the presence of a single normal allele is sufficient to prevent bleeding, making CalDAG-GEFI a novel and potentially safe therapeutic target to prevent thrombosis. PMID:24958846

  8. A DAG Scheduling Scheme on Heterogeneous Computing Systems Using Tuple-Based Chemical Reaction Optimization

    PubMed Central

    Jiang, Yuyi; Shao, Zhiqing; Guo, Yi

    2014-01-01

    A complex computing problem can be solved efficiently on a system with multiple computing nodes by dividing its implementation code into several parallel processing modules or tasks that can be formulated as directed acyclic graph (DAG) problems. The DAG jobs may be mapped to and scheduled on the computing nodes to minimize the total execution time. Searching an optimal DAG scheduling solution is considered to be NP-complete. This paper proposed a tuple molecular structure-based chemical reaction optimization (TMSCRO) method for DAG scheduling on heterogeneous computing systems, based on a very recently proposed metaheuristic method, chemical reaction optimization (CRO). Comparing with other CRO-based algorithms for DAG scheduling, the design of tuple reaction molecular structure and four elementary reaction operators of TMSCRO is more reasonable. TMSCRO also applies the concept of constrained critical paths (CCPs), constrained-critical-path directed acyclic graph (CCPDAG) and super molecule for accelerating convergence. In this paper, we have also conducted simulation experiments to verify the effectiveness and efficiency of TMSCRO upon a large set of randomly generated graphs and the graphs for real world problems. PMID:25143977

  9. Quantification of TAG and DAG in lesquerella (Physaria fendleri) oil by HPLC and MS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Castor oil has many industrial uses because of its high content (90%) of the hydroxy fatty acid, ricinoleic acid (OH1218:19). Lesquerella oil containing lesquerolic acid (Ls, OH1420:111, 56.5%) is potentially useful in industry. Ten diacylglycerols (DAG) and 74 triacylglycerols (TAG) in the seed oil...

  10. DagStream: locality aware and failure resilient peer-to-peer streaming

    NASA Astrophysics Data System (ADS)

    Liang, Jin; Nahrstedt, Klara

    2006-01-01

    Live peer to peer (P2P) media streaming faces many challenges such as peer unreliability and bandwidth heterogeneity. To effectively address these challenges, general "mesh" based P2P streaming architectures have recently been adopted. Mesh-based systems allow peers to aggregate bandwidth from multiple neighbors, and dynamically adapt to changing network conditions and neighbor failures. However, a drawback of mesh-based overlays is that it is difficult to guarantee network connectivity in a distributed fashion, especially when network locality needs to be optimized. This paper introduces a new P2P streaming framework called DagStream, which (1) organizes peers into a directed acyclic graph (DAG) where each node maintains at least k parents, thus has provable network connectivity (and hence failure resilience), and (2) enables peers to quickly achieve locality awareness in a distributed fashion, thus ensures efficient network resource usage. Our experiment results in both simulation and wide area environment show that with our DagStream protocol, peers can quickly self-organize into a locality aware DAG. Further, by selecting additional parents as needed, peers can achieve good streaming quality commensurate with their downlink bandwidth.

  11. Tyrosinase kinetics in epidermal melanocytes: analysis of DAG-PKC-dependent signaling pathway

    NASA Astrophysics Data System (ADS)

    Stolnitz, Mikhail M.; Peshkova, Anna Y.

    2001-05-01

    Tyrosinase is the key enzyme of melanogenesis with unusual enzyme kinetics. Protein kinase C plays an important role in regulating of tyrosinase activity. In the paper the mathematical model of PKC-DAG-dependent signal transduction pathway for UV-radiation is presented.

  12. Design and synthesis of protein kinase C epsilon selective diacylglycerol lactones (DAG-lactones).

    PubMed

    Ann, Jihyae; Yoon, Suyoung; Baek, Jisoo; Kim, Da Hye; Lewin, Nancy E; Hill, Colin S; Blumberg, Peter M; Lee, Jeewoo

    2015-01-27

    DAG-lactones afford a synthetically accessible, high affinity platform for probing structure activity relationships at the C1 regulatory domain of protein kinase C (PKC). Given the central role of PKC isoforms in cellular signaling, along with their differential biological activities, a critical objective is the design of isoform selective ligands. Here, we report the synthesis of a series of DAG-lactones varying in their side chains, with a particular focus on linoleic acid derivatives. We evaluated their selectivity for PKC epsilon versus PKC alpha both under standard lipid conditions (100% phosphatidylserine, PS) as well as in the presence of a nuclear membrane mimetic lipid mixture (NML). We find that selectivity for PKC epsilon versus PKC alpha tended to be enhanced in the presence of the nuclear membrane mimetic lipid mixture and, for our lead compound, report a selectivity of 32-fold. PMID:25437619

  13. Defining order and timing of mutations during cancer progression: the TO-DAG probabilistic graphical model

    PubMed Central

    Lecca, Paola; Casiraghi, Nicola; Demichelis, Francesca

    2015-01-01

    Somatic mutations arise and accumulate both during tumor genesis and progression. However, the order in which mutations occur is an open question and the inference of the temporal ordering at the gene level could potentially impact on patient treatment. Thus, exploiting recent observations suggesting that the occurrence of mutations is a non-memoryless process, we developed a computational approach to infer timed oncogenetic directed acyclic graphs (TO-DAGs) from human tumor mutation data. Such graphs represent the path and the waiting times of alterations during tumor evolution. The probability of occurrence of each alteration in a path is the probability that the alteration occurs when all alterations prior to it have occurred. The waiting time between an alteration and the subsequent is modeled as a stochastic function of the conditional probability of the event given the occurrence of the previous one. TO-DAG performances have been evaluated both on synthetic data and on somatic non-silent mutations from prostate cancer and melanoma patients and then compared with those of current well-established approaches. TO-DAG shows high performance scores on synthetic data and recognizes mutations in gatekeeper tumor suppressor genes as trigger for several downstream mutational events in the human tumor data. PMID:26528329

  14. Des-acyl-ghrelin (DAG) normalizes hyperlactacidemia and improves survival in a lethal rat model of burn trauma.

    PubMed

    Sheriff, Sulaiman; Kadeer, Nijiati; Friend, Lou Ann; James, J Howard; Alexander, J Wesley; Balasubramaniam, Ambikaipakan

    2014-10-01

    Critical illness, including burn injury, results in elevated plasma lactate levels. Dysregulation of PI3K/Akt signaling has been shown to play a predominant role in the inactivation of skeletal muscle PDC and, hence, in hyperlactacidemia in rat models of sepsis and endotoxemia. This observation, and our previous finding that DAG can reverse burn-induced skeletal muscle proteolysis through the activation of PI3K/Akt pathway, led us to hypothesize that DAG may also attenuate hyperlactacidemia in burn injury. Our investigations revealed that burn injury significantly elevated both skeletal muscle lactate production and plasma lactate levels. Moreover, this was accompanied in skeletal muscle by a 5-7 fold increase in mRNA expression of pyruvate dehydrogenase kinases (PDK) 2 and 4, and a ∼30% reduction in PDC activity. DAG treatment of burn rats completely normalized not only the mRNA expression of the PDKs and PDC activity, but also hyperlactacidemia within 24h of burn injury. DAG also normalized epinephrine-induced lactate production by isolated skeletal muscles from normal rats. Moreover, DAG also improved survival in a lethal rat model of burn trauma. These findings with DAG may have clinical implications because chances of survival for critically ill patients are greatly improved if plasma lactate levels are normalized within 24h of injury. PMID:25063053

  15. Nemrud Dag

    NASA Astrophysics Data System (ADS)

    Belmonte, Juan Antonio; González-García, A. César

    The World Heritage Site of the hierothesion of Antiochos I, King of Commagene, at Mount Nemrud (Turkey) certainly constitutes one of the most fascinating historical enigmas in human culture. The monument includes the famous lion "horoscope" which has often been used in various attempts to interpret and date the ruins with controversial results. According to recent analyses, Antiochos' monument reflects the situation of the skies at exclusive moments of the year 49 BC, when the monument would have been started. This alternative explanation considers the lion slab together with the orientation of the eastern and western terraces of the hierothesion and the inscriptions on the monument.

  16. Biochemical Properties and Structure Analysis of a DAG-Like Lipase from Malassezia globosa

    PubMed Central

    Xu, Huan; Lan, Dongming; Yang, Bo; Wang, Yonghua

    2015-01-01

    Diacylglycerol (DAG)-like lipases are found to play an important role in the life sciences and industrial fields. A putative DAG-like lipase (MgMDL2) from Malassezia globosa was cloned and expressed in recombinant Pichia pastoris. The recombinant MgMDL2 was expressed as a glycosylated protein and purified into homogeneity by anion exchange chromatography. The activity of recombinant MgMDL2 was optimal at 15 °C and pH 6.0, and it keeps over 50% of relative activity at 5 °C, suggesting that MgMDL2 was a cold active lipase. MgMDL2 retained over 80% of initial activity after incubation at 30 and 40 °C for 2.5 h, but it was not stable at 50 °C. Incubation of methanol and ethanol at a concentration of 30% for 2 h did not affect the recombinant enzyme activity, while metal ions, including Ca2+, Mn2+ and Ni2+, sharply inhibited the MgMDL2 activity at 5 mM by 42%, 35% and 36%, respectively. MgMDL2 exhibited a preference for medium chain-length esters with highest activity toward p-nitrophenyl caprylate, while it was active on mono- and diacylglycerol but not on triacylglycerol, indicating that it was a typical DAG-like lipase. By homology modeling, Phe278 was predicted to be involved in the preference of MgMDL2 for monoacyl- and diacyl-glyceride substrates, but not triglycerides. PMID:25749469

  17. Diagnostic Analyzer for Gearboxes (DAG): User's Guide. Version 3.1 for Microsoft Windows 3.1

    NASA Technical Reports Server (NTRS)

    Jammu, Vinay B.; Kourosh, Danai

    1997-01-01

    This documentation describes the Diagnostic Analyzer for Gearboxes (DAG) software for performing fault diagnosis of gearboxes. First, the user would construct a graphical representation of the gearbox using the gear, bearing, shaft, and sensor tools contained in the DAG software. Next, a set of vibration features obtained by processing the vibration signals recorded from the gearbox using a signal analyzer is required. Given this information, the DAG software uses an unsupervised neural network referred to as the Fault Detection Network (FDN) to identify the occurrence of faults, and a pattern classifier called Single Category-Based Classifier (SCBC) for abnormality scaling of individual vibration features. The abnormality-scaled vibration features are then used as inputs to a Structure-Based Connectionist Network (SBCN) for identifying faults in gearbox subsystems and components. The weights of the SBCN represent its diagnostic knowledge and are derived from the structure of the gearbox graphically presented in DAG. The outputs of SBCN are fault possibility values between 0 and 1 for individual subsystems and components in the gearbox with a 1 representing a definite fault and a 0 representing normality. This manual describes the steps involved in creating the diagnostic gearbox model, along with the options and analysis tools of the DAG software.

  18. A novel DAG-dependent mechanism links PKCa and Cyclin B1 regulating cell cycle progression

    PubMed Central

    Poli, Alessandro; Ramazzotti, Giulia; Matteucci, Alessandro; Manzoli, Lucia; Lonetti, Annalisa; Suh, Pann-Ghill; McCubrey, James A.; Cocco, Lucio

    2014-01-01

    Through the years, different studies showed the involvement of Protein Kinase C (PKC) in cell cycle control, in particular during G1/S transition. Little is known about their role at G2/M checkpoint. In this study, using K562 human erythroleukemia cell line, we found a novel and specific mechanism through which the conventional isoform PKC⍺ positively affects Cyclin B1 modulating G2/M progression of cell cycle. Since the kinase activity of this PKC isoform was not necessary in this process, we demonstrated that PKC⍺, physically interacting with Cyclin B1, avoided its degradation and stimulated its nuclear import at mitosis. Moreover, the process resulted to be strictly connected with the increase in nuclear diacylglycerol levels (DAG) at G2/M checkpoint, due to the activity of nuclear Phospholipase C β1 (PLCβ1), the only PLC isoform mainly localized in the nucleus of K562 cells. Taken together, our findings indicated a novel DAG dependent mechanism able to regulate the G2/M progression of the cell cycle. PMID:25362646

  19. Fast synthesis of 1,3-DAG by Lecitase® Ultra-catalyzed esterification in solvent-free system

    PubMed Central

    Liu, Ning; Wang, Yong; Zhao, Qiangzhong; Zhang, Qingli; Zhao, Mouming

    2011-01-01

    Lecitase® Ultra, a phospholipase, was explored as an effective biocatalyst for direct esterification of glycerol with oleic acid to produce 1,3-DAG. Experiments were carried out in batch mode, and optimal reaction conditions were evaluated. In comparison with several organic solvent mediums, the solvent-free system was found to be more beneficial for this esterification reaction, which was further studied to investigate the reaction conditions including oleic acid/glycerol mole ratio, temperature, initial water content, enzyme load, and operating time. The results showed that Lecitase® Ultra catalyzed a fast synthesis of 1,3-DAG by direct esterification in a solvent-free medium. Under the optimal reaction conditions, a short reaction time 1.5 h was found to achieve the fatty acid esterification efficiency of 80.3 ± 1.2% and 1,3-DAG content of 54.8 ± 1.6 wt% (lipid layer of reaction mixture mass). The reusability of Lecitase® Ultra was evaluated via recycling the excess glycerol layer in the reaction system. DAG in the upper lipid layer of reaction mixture was purified by molecular distillation and the 1,3-DAG-enriched oil with a purity of about 75 wt% was obtained. Practical applications: The new Lecitase® Ultra catalyzed process for production of 1,3-DAG from glycerol and oleic acid described in this study provides several advantages over conventional methods including short reaction time, the absence of a solvents and a high product yield. PMID:21966255

  20. Fast synthesis of 1,3-DAG by Lecitase® Ultra-catalyzed esterification in solvent-free system.

    PubMed

    Liu, Ning; Wang, Yong; Zhao, Qiangzhong; Zhang, Qingli; Zhao, Mouming

    2011-08-01

    Lecitase® Ultra, a phospholipase, was explored as an effective biocatalyst for direct esterification of glycerol with oleic acid to produce 1,3-DAG. Experiments were carried out in batch mode, and optimal reaction conditions were evaluated. In comparison with several organic solvent mediums, the solvent-free system was found to be more beneficial for this esterification reaction, which was further studied to investigate the reaction conditions including oleic acid/glycerol mole ratio, temperature, initial water content, enzyme load, and operating time. The results showed that Lecitase® Ultra catalyzed a fast synthesis of 1,3-DAG by direct esterification in a solvent-free medium. Under the optimal reaction conditions, a short reaction time 1.5 h was found to achieve the fatty acid esterification efficiency of 80.3 ± 1.2% and 1,3-DAG content of 54.8 ± 1.6 wt% (lipid layer of reaction mixture mass). The reusability of Lecitase® Ultra was evaluated via recycling the excess glycerol layer in the reaction system. DAG in the upper lipid layer of reaction mixture was purified by molecular distillation and the 1,3-DAG-enriched oil with a purity of about 75 wt% was obtained.Practical applications: The new Lecitase® Ultra catalyzed process for production of 1,3-DAG from glycerol and oleic acid described in this study provides several advantages over conventional methods including short reaction time, the absence of a solvents and a high product yield. PMID:21966255

  1. On the Feasibility of Dynamically Scheduling DAG Applications on Shared Heterogeneous Systems

    NASA Astrophysics Data System (ADS)

    Nascimento, Aline P.; Sena, Alexandre; Boeres, Cristina; Rebello, Vinod E. F.

    Grid and Internet Computing have proved their worth executing large-scale bag-of-task class applications. Numerous middlewares have been developed to manage their execution in either dedicated environments or opportunistic and shared ad-hoc grids. While job dependencies are now being resolved by middleware capable of scheduling workflows, these environments have yet to be shown beneficial for message passing parallel applications. Obtaining high performance in these widely available environments without rewriting existing parallel applications is of up most importance to e-Science. The key to an efficient solution may be an alternative execution model and the efficient dynamic scheduling of application processes. This paper presents a hierarchical scheme for dynamically scheduling parallel DAG applications across a set of non-dedicated heterogeneous resources. In order to efficiently tackle process dependencies and adapt to varying system characteristics, dynamic schedulers are distributed within the application and operate in a collaborative and pro-active fashion to keep overheads low.

  2. Study of the recent geodynamic processes in the Kopet-Dag region

    NASA Astrophysics Data System (ADS)

    Izyumov, S. F.; Kuzmin, Yu. O.

    2014-11-01

    The time series of the uniquely long geodetic observations of recent geodynamic processes in the Kopet-Dag region are analyzed. The regional observations of contemporary vertical movements cover a 75-year period; the zonal and local systems of leveling measurements, which provide an increased degree of spatiotemporal detail (the distance between the benchmarks is less than 1 km and the measurements are repeated with a frequency of once per month to two times per annum), have been functioning for 50 years. It is shown that during the last 40-50 years, the regional stress field in the forefront of the Main Kopet-Dag thrust and collision zone of the Turanian and Iranian plates is quasi static. The annual average trend rate of strains estimated from a set of the time series of uniquely long geodetic observations is (3-5) × 10-8 yr-1, which is only one to two amplitudes of tidal deformations of the solid Earth. The local deformations in the fault zone reach the rates that are by 1.5-2 orders of magnitude higher than in the block part of the region. It is found that the segments of the Earth's surface within the axial part of the depression experience persistent uplifting, which indicates that they do not follow the scheme of inherited evolution characteristic of the fault-block structures of the region. It is demonstrated that these anomalous uplifts can be caused by the variations in the weak seismicity in the zone of the North Ashgabat Fault.

  3. Enzymatic selective synthesis of 1,3-DAG based on deep eutectic solvent acting as substrate and solvent.

    PubMed

    Zeng, Chao-Xi; Qi, Sui-Jian; Xin, Rui-Pu; Yang, Bo; Wang, Yong-Hua

    2015-11-01

    In this study, enzymatic selective esterification of oleic acid with glycerol based on deep eutectic solvent acting as substrate and solvent was studied. As choline chloride (ChCl) or betaine can effectively change the chemical reaction characteristics of glycerol when they are mixed with a certain molar ratio of glycerol, several factors crucial to the lipase catalytic esterification of glycerol with oleic acid was investigated. Results showed that, betaine had more moderate effects than ChCl on the lipase, and water content had an important influence of the esterification and the enzyme selectivity. Significant changes of the glyceride compositions and enzyme selectivity were found in ChCl adding system compared with pure glycerol system; optimum accumulation of DAG especially 1,3-DAG because of the eutectic effect of ChCl was found in this system. Furthermore, in a model 1,3-DAG esterification synthesis system catalyzed by Novozym 435, high content (42.9 mol%) of the 1,3-DAG could be obtained in ChCl adding system within 1 h. PMID:26210852

  4. Fostered to Internationalism and Peace: Biographical Notes on UN General Secretary Dag Hammarskjold. Peace Education Miniprints No. 97.

    ERIC Educational Resources Information Center

    Thelin, Bengt

    This paper presents some biographical glimpses of Dag Hammarskjold's childhood and youth. It is a period of the life of the late United Nations (UN) Secretary General (1905-1961) that has been dealt with very little by his biographers. Hammarskjold's private archives at the Royal Library in Stockholm are now available for research. These sources…

  5. Novel mutations in RASGRP2, which encodes CalDAG-GEFI, abrogate Rap1 activation, causing platelet dysfunction.

    PubMed

    Lozano, María Luisa; Cook, Aaron; Bastida, José María; Paul, David S; Iruin, Gemma; Cid, Ana Rosa; Adan-Pedroso, Rosa; Ramón González-Porras, José; Hernández-Rivas, Jesús María; Fletcher, Sarah J; Johnson, Ben; Morgan, Neil; Ferrer-Marin, Francisca; Vicente, Vicente; Sondek, John; Watson, Steve P; Bergmeier, Wolfgang; Rivera, José

    2016-09-01

    In addition to mutations in ITG2B or ITGB3 genes that cause defective αIIbβ3 expression and/or function in Glanzmann's thrombasthenia patients, platelet dysfunction can be a result of genetic variability in proteins that mediate inside-out activation of αIIbβ3 The RASGRP2 gene is strongly expressed in platelets and neutrophils, where its encoded protein CalDAG-GEFI facilitates the activation of Rap1 and subsequent activation of integrins. We used next-generation sequencing (NGS) and whole-exome sequencing (WES) to identify 2 novel function-disrupting mutations in RASGRP2 that account for bleeding diathesis and platelet dysfunction in 2 unrelated families. By using a panel of 71 genes, we identified a homozygous change (c.1142C>T) in exon 10 of RASGRP2 in a 9-year-old child of Chinese origin (family 1). This variant led to a p.Ser381Phe substitution in the CDC25 catalytic domain of CalDAG-GEFI. In 2 Spanish siblings from family 2, WES identified a nonsense homozygous variation (c.337C>T) (p.Arg113X) in exon 5 of RASGRP2 CalDAG-GEFI expression was markedly reduced in platelets from all patients, and by using a novel in vitro assay, we found that the nucleotide exchange activity was dramatically reduced in CalDAG-GEFI p.Ser381Phe. Platelets from homozygous patients exhibited agonist-specific defects in αIIbβ3 integrin activation and aggregation. In contrast, α- and δ-granule secretion, platelet spreading, and clot retraction were not markedly affected. Integrin activation in the patients' neutrophils was also impaired. These patients are the first cases of a CalDAG-GEFI deficiency due to homozygous RASGRP2 mutations that are linked to defects in both leukocyte and platelet integrin activation. PMID:27235135

  6. 77 FR 38391 - Mercedes-Benz USA, LLC, and Daimler AG (DAG), Receipt of Petition for Decision of Inconsequential...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-06-27

    ...Mercedes-Benz USA, LLC \\1\\ (MBUSA) and its parent company Daimler AG (DAG)(collectively referred to as ``MB'') have determined that certain model year 2011 and 2012 Mercedes-Benz S-Class (221 platform) passenger cars do not fully comply with paragraph S4.4 TPMS Malfunction of Federal Motor Vehicle Safety Standard (FMVSS) No. 138, Tire Pressure Monitoring Systems. MB has filed an appropriate......

  7. Martian Pyroxenes in the Shergottite Meteorites; Zagami, SAU005, DAG476 and EETA79001

    NASA Astrophysics Data System (ADS)

    Stephen, N.; Benedix, G. K.; Bland, P.; Hamilton, V. E.

    2010-12-01

    The geology and surface mineralogy of Mars is characterised using remote sensing techniques such as thermal emission spectroscopy (TES) from instruments on a number of spacecraft currently orbiting Mars or gathered from roving missions on the Martian surface. However, the study of Martian meteorites is also important in efforts to further understand the geological history of Mars or to interpret mission data as they are believed to be the only available samples that give us direct clues as to Martian igneous processes [1]. We have recently demonstrated that the spectra of Martian-specific minerals can be determined using micro-spectroscopy [2] and that these spectra can be reliably obtained from thin sections of Martian meteorites [3]. Accurate modal mineralogy of these meteorites is also important [4]. In this study we are using a variety of techniques to build upon previous studies of these particular samples in order to fully characterise the nature of the 2 common pyroxenes found in Martian Shergottites; pigeonite and augite [5], [6]. Previous studies have shown that the Shergottite meteorites are dominated by pyroxene (pigeonite and augite in varying quantities) [4], [5], commonly but not always olivine, plagioclase or maskelynite/glass and also hydrous minerals, which separate the Martian meteorites from other achondrites [7]. Our microprobe study of meteorites Zagami, EETA79001, SAU005 and DAG476 in thin-section at the Natural History Museum, London shows a chemical variability within both the pigeonite and augite composition across individual grains in all thin sections; variation within either Mg or Ca concentration varies from core to rim within the grains. This variation can also be seen in modal mineralogy maps using SEM-derived element maps and the Photoshop® technique previously described [4], and in new micro-spectroscopy data, particularly within the Zagami meteorite. New mineral spectra have been gathered from the Shergottite thin-sections by

  8. A new monoclonal antibody DAG-6F4 against human alpha-dystroglycan reveals reduced core protein in some, but not all, dystroglycanopathy patients.

    PubMed

    Humphrey, Emma L; Lacey, Erica; Le, Lam T; Feng, Lucy; Sciandra, Francesca; Morris, Charlotte R; Hewitt, Jane E; Holt, Ian; Brancaccio, Andrea; Barresi, Rita; Sewry, Caroline A; Brown, Susan C; Morris, Glenn E

    2015-01-01

    We generated a novel monoclonal antibody, DAG-6F4, against alpha-dystroglycan which immunolabels the sarcolemma in human muscle biopsies. Its seven amino-acid epitope, PNQRPEL, was identified using phage-displayed peptides and is located immediately after the highly-glycosylated mucin domain of alpha-dystroglycan. On Western blots of recombinant alpha-dystroglycan, epitope accessibility was reduced, but not entirely prevented, by glycosylation. DAG-6F4 immunolabelling was markedly reduced in muscle biopsies from Duchenne muscular dystrophy patients consistent with disruption of the dystroglycan complex. In a range of dystroglycanopathy patients with reduced/altered glycosylation, staining by DAG-6F4 was often less reduced than staining by IIH6 (antibody against the glycan epitope added by LARGE and commonly used to identify glycosylated alpha-dystroglycan). Whereas IIH6 was reduced in all patients, DAG-6F4 was hardly changed in a LARGE patient, less reduced than IIH6 in limb-girdle muscular dystrophy type 2I, but as reduced as IIH6 in some congenital muscular dystrophy patients. Although absence of the LARGE-dependent laminin-binding site appears not to affect alpha-dystroglycan stability at the sarcolemma, the results suggest that further reduction in aDG glycosylation may reduce its stability. These studies suggest that DAG-6F4 may be a useful addition to the antibody repertoire for evaluating the dystroglycan complex in neuromuscular disorders. PMID:25387694

  9. Inactivation of the Phloem-Specific Dof Zinc Finger Gene DAG1 Affects Response to Light and Integrity of the Testa of Arabidopsis Seeds1

    PubMed Central

    Papi, Maura; Sabatini, Sabrina; Altamura, Maria Maddalena; Hennig, Lars; Schäfer, Eberhard; Costantino, Paolo; Vittorioso, Paola

    2002-01-01

    We show here that seeds from the knockout mutant of the Arabidopsis DAG1 gene encoding a Dof zinc finger transcription factor have an altered response to red and far-red light. Mutant dag1 seeds are induced to germinate by much lower red light fluence rates, and germination reaches more quickly a point where it is independent of phytochrome signaling. Moreover, although microscopic analysis reveals no obvious structural alterations in the seed coat (testa) of dag1 seeds, staining assays with different dyes point to an abnormal fragility of the testa. By extensive in situ mRNA hybridization analysis we show here that the gene, which is not expressed in the embryo, is specifically expressed in the phloem of all organs of the mother plant. PMID:11842145

  10. Synthesis, biological, and biophysical studies of DAG-indololactones designed as selective activators of RasGRP.

    PubMed

    Garcia, Lia C; Donadío, Lucia Gandolfi; Mann, Ella; Kolusheva, Sofiya; Kedei, Noemi; Lewin, Nancy E; Hill, Colin S; Kelsey, Jessica S; Yang, Jing; Esch, Timothy E; Santos, Marina; Peach, Megan L; Kelley, James A; Blumberg, Peter M; Jelinek, Raz; Marquez, Victor E; Comin, Maria J

    2014-06-15

    The development of selective agents capable of discriminating between protein kinase C (PKC) isoforms and other diacylglycerol (DAG)-responsive C1 domain-containing proteins represents an important challenge. Recent studies have highlighted the role that Ras guanine nucleotide-releasing protein (RasGRP) isoforms play both in immune responses as well as in the development of prostate cancer and melanoma, suggesting that the discovery of selective ligands could have potential therapeutic value. Thus far, the N-methyl-substituted indololactone 1 is the agonist with the highest reported potency and selectivity for RasGRP relative to PKC. Here we present the synthesis, binding studies, cellular assays and biophysical analysis of interactions with model membranes of a family of regioisomers of 1 (compounds 2-5) that differ in the position of the linkage between the indole ring and the lactone moiety. These structural variations were studied to explore the interaction of the active complex (C1 domain-ligand) with cellular membranes, which is believed to be an important factor for selectivity in the activation of DAG-responsive C1 domain containing signaling proteins. All compounds were potent and selective activators of RasGRP when compared to PKCα with selectivities ranging from 6 to 65 fold. However, the parent compound 1 was appreciably more selective than any of the other isomers. In intact cells, modest differences in the patterns of translocation of the C1 domain targets were observed. Biophysical studies using giant vesicles as model membranes did show substantial differences in terms of molecular interactions impacting lipid organization, dynamics and membrane insertion. However, these differences did not yield correspondingly large changes in patterns of biological response, at least for the parameters examined. PMID:24794745

  11. Synthesis, Biological, and Biophysical Studies of DAG-indololactones Designed as Selective Activators of RasGRP

    PubMed Central

    Garcia, Lia C.; Donadío, Lucia Gandolfi; Mann, Ella; Kolusheva, Sofiya; Kedei, Noemi; Lewin, Nancy E.; Hill, Colin S.; Kelsey, Jessica S.; Yang, Jing; Esch, Timothy E.; Santos, Marina; Peach, Megan L.; Kelley, James A.; Blumberg, Peter M.; Jelinek, Raz; Marquez, Victor E.; Comin, Maria J.

    2014-01-01

    The development of selective agents capable of discriminating between protein kinase C (PKC) isoforms and other diacylglycerol (DAG)-responsive C1 domain-containing proteins represents an important challenge. Recent studies have highlighted the role that Ras guanine nucleotide-releasing protein (RasGRP) isoforms play both in immune responses as well as in the development of prostate cancer and melanoma, suggesting that the discovery of selective ligands could have potential therapeutic value. Thus far, the N-methyl-substituted indololactone 1 is the agonist with the highest reported potency and selectivity for RasGRP relative to PKC. Here we present the synthesis, binding studies, cellular assays and biophysical analysis of interactions with model membranes of a family of regioisomers of 1 (compounds 2 to 5) that differ in the position of the linkage between the indole ring and the lactone moiety. These structural variations were studied to explore the interaction of the active complex (C1 domain-ligand) with cellular membranes, which is believed to be an important factor for selectivity in the activation of DAG-responsive C1 domain containing signaling proteins. All compounds were potent and selective activators of RasGRP when compared to PKCα with selectivities ranging from 6 to 65 fold. However, the parent compound 1 was appreciably more selective than any of the other isomers. In intact cells, modest differences in the patterns of translocation of the C1 domain targets were observed. Biophysical studies using giant vesicles as model membranes did show substantial differences in terms of molecular interactions impacting lipid organization, dynamics and membrane insertion. However, these differences did not yield correspondingly large changes in patterns of biological response, at least for the parameters examined. PMID:24794745

  12. The Story of a Seminar in Applied Communication. The Dag Hammarskjold Seminar on "Communication--An Essential Component in Development Work" (Uppsala, Sweden, August 27-September 9, 1972).

    ERIC Educational Resources Information Center

    Fuglesang, Andreas, Ed.

    The objective of the two week long 1972 Dag Hammarskjold Seminar was to break away from traditional and theoretical concepts of information work in order to gain a practical insight into communication, especially as it applies to social and economic development. Fifty participants and lecturers from developed and developing countries in Europe and…

  13. Reduced expression of CDP-DAG synthase changes lipid composition and leads to male sterility in Drosophila

    PubMed Central

    Laurinyecz, Barbara; Péter, Mária; Vedelek, Viktor; Kovács, Attila L.; Juhász, Gábor; Maróy, Péter; Vígh, László; Balogh, Gábor; Sinka, Rita

    2016-01-01

    Drosophila spermatogenesis is an ideal system to study the effects of changes in lipid composition, because spermatid elongation and individualization requires extensive membrane biosynthesis and remodelling. The bulk of transcriptional activity is completed with the entry of cysts into meiotic division, which makes post-meiotic stages of spermatogenesis very sensitive to even a small reduction in gene products. In this study, we describe the effect of changes in lipid composition during spermatogenesis using a hypomorphic male sterile allele of the Drosophila CDP-DAG synthase (CdsA) gene. We find that the CdsA mutant shows defects in spermatid individualization and enlargement of mitochondria and the axonemal sheath of the spermatids. Furthermore, we could genetically rescue the male sterile phenotype by overexpressing Phosphatidylinositol synthase (dPIS) in a CdsA mutant background. The results of lipidomic and genetic analyses of the CdsA mutant highlight the importance of correct lipid composition during sperm development and show that phosphatidic acid levels are crucial in late stages of spermatogenesis. PMID:26791243

  14. Mineralogical and Geochemical Analysis of Howardite DaG 779: understanding geological evolution of asteroid (4) Vesta

    NASA Astrophysics Data System (ADS)

    Marcel Müller, Christian; Mengel, Kurt; Singh Thangjam, Guneshwar; Weckwerth, Gerd

    2016-04-01

    The HED meteorites, a clan of stony achondrites, are believed to originate from asteroid (4) Vesta (e.g. Mittlefehldt et al. (2015)). Recent evolution models (e.g. Toplis et al. (2013)) and observations from Dawn spacecraft data (e.g., Prettyman et al. (2013)) indicate that diogenites form the lower crust and uppermost mantle of (4) Vesta. Deep seated material excavated by large impacts such as the Rheasilvia- and Veneneiaforming event should be present in howardites. We analysed a slice of howardite DaG 779 which had been recovered from the Libyan Desert in 1999 and was briefly described by Grossmann (2000). The data presented here include electron microprobe, bulk-rock XRD and XRF as well as trace element analysis by ICP-MS and INA. The petrographic results confirm earlier observations that DaG 779 is polymict and mainly contains diogenite and eucrite clasts. Mass balance calculations using bulk-rock and microprobe major element data reveal a modal mineralogy of 77% orthopyroxene, 8% plagioclase, 7% clinopyroxene and 2% spinels, the rest being olivine, SiO2-phases, sulphides, and native Fe(Ni). When compared with the element compilation recently reported by Mittlefehldt (2015) the 39 trace element analysed here (including REE and PGE) confirm that this howardite is clearly dominated by diogenite. Beside the modal petrographic information, a number of more detailed observations obtained from microprobe investigations reveal fresh and recrystallized glasses, troilite-orthopyroxene symplectites from a mixed silicate-sulphide melt giving rise to graphic intergrowths as well as vermicular and reticular FeS in highly disrupted clasts. While the origin of the FeS in these clasts is not clear yet, its particular shape and distribution indicates that this mineral has been (partially) molten and recrystallized from a sulphide melt. The silicate minerals around these FeS occurrences are recrystallized but there is no indication for a partial silicate melt. Further

  15. Airborne Conflict Management within Confined Airspace in a Piloted Simulation of DAG-TM Autonomous Aircraft Operations

    NASA Technical Reports Server (NTRS)

    Barmore, Bryan; Johnson, Edward; Wing, David J.; Barhydt, Richard

    2003-01-01

    A human-in-the-loop experiment was performed at the NASA Langley Research Center to study the feasibility of Distributed Air/Ground Traffic Management (DAG-TM) autonomous aircraft operations in highly constrained airspace. The airspace was constrained by a pair of special use airspace (SUA) regions on either side of the pilot s planned route. The available airspace was further varied by changing the separation standard for lateral separation between 3 nm and 5 nm. The pilot had to maneuver through the corridor between the SUA s, avoid other traffic and meet flow management constraints. Traffic flow management (TFM) constraints were imposed as a required time of arrival and crossing altitude at an en route fix. This is a follow-up study to work presented at the 4th USA/Europe Air Traffic Management R&D Seminar in December 2001. Nearly all of the pilots were able to meet their TFM constraints while maintaining adequate separation from other traffic. In only 3 out of 59 runs were the pilots unable to meet their required time of arrival. Two loss of separation cases are studied and it is found that the pilots need conflict prevention information presented in a clearer manner. No degradation of performance or safety was seen between the wide and narrow corridors. Although this was not a thorough study of the consequences of reducing the en route lateral separation, nothing was found that would refute the feasibility of reducing the separation requirement from 5 nm to 3 nm. The creation of additional, second-generation conflicts is also investigated. Two resolution methods were offered to the pilots: strategic and tactical. The strategic method is a closed-loop alteration to the Flight Management System (FMS) active route that considers other traffic as well as TFM constraints. The tactical resolutions are short-term resolutions that leave avoiding other traffic conflicts and meeting the TFM constraints to the pilot. Those that made use of the strategic tools avoided

  16. Conformationally Constrained Analogues of Diacylglycerol (DAG). 25. Exploration of the sn-1 and sn-2 carbonyl functionality reveals the essential role of the sn-1 carbonyl at the lipid interface in the binding of DAG-lactones to protein kinase C

    PubMed Central

    Kang, Ji-Hye; Peach, Megan L.; Pu, Yongmei; Lewin, Nancy E.; Nicklaus, Marc C.; Blumberg, Peter M.; Marquez, Victor E.

    2008-01-01

    A group of DAG-lactones with altered functionality (C=O → CH2 or C=O → C=S) at the sn-1 and sn-2 carbonyl pharmacophores was synthesized and used as probes to dissect the individual role of each carbonyl in binding to protein kinase C (PKC). The results suggest that the hydrated sn-1 carbonyl is engaged in very strong hydrogen bonding interactions with the charged lipid headgroups and organized water molecules at the lipid interface. Conversely, the sn-2 carbonyl has a more modest contribution to the binding process as a result of its involvement with the receptor (C1 domain) via conventional hydrogen bonding to the protein. The parent DAG-lactones, E-6 and Z-7, were designed to bind exclusively in the sn-2 binding mode to insure the correct orientation and disposition of pharmacophores at the binding site. PMID:16134942

  17. A functional interaction between TRPC/NCKX induced by DAG plays a role in determining calcium influx independently from PKC activation.

    PubMed

    Pulcinelli, Fabio M; Trifirò, Elisabetta; Massimi, Isabella; Di Renzo, Livia

    2013-01-01

    Ca(2+)influx might occur through K(+)-dependent Na(+)/Ca(2+) exchanger operating in reverse mode (rNCKX). In a cellular model different from platelets, an interaction between canonical transient receptor potential cation (TRPC) channels and NCX has been found. The aim of this study was to verify whether the TRPC/NCKX interaction operates in human platelets. Our results showed that the diacylglycerol (DAG) analogue, 1-oleoyl-2-acetyl-sn-glycerol (OAG) induced rNCKX-mediated Ca(2+) influx through TRPC-mediated Na(+) influx. DAG-induced activation of TRPC/NCKX occurs independently of protein kinase C (PKC) activation, as PKC inhibitor did not modify OAG-mediated Ca(2+) influx. Moreover, as both rNCKX and TRPC inhibitors reduced OAG-induced platelet aggregation which, conversely, was increased by flufenamic acid, known to develop TRPC activity, it could be suggested that the TRPC/NCKX interaction has a role in OAG-dependent platelet aggregation. PMID:23249278

  18. Duplex-tetraplex equilibrium between a hairpin and two interacting hairpins of d(A-G)10 at neutral pH.

    PubMed Central

    Shiber, M C; Braswell, E H; Klump, H; Fresco, J R

    1996-01-01

    d(A-G)10 forms two helical structures at neutrality, at low ionic strength a single-hairpin duplex, and at higher ionic strength a double-hairpin tetraplex. An ionic strength-dependent equilibrium between these forms is indicated by native PAGE, which also reveals additional single-stranded species below 0.3 M Na+, probably corresponding to partially denatured states. The equilibrium also depends upon oligomer concentration: at very low concentrations, d(A-G)10 migrates faster than the random coil d(C-T)10, probably because it is a more compact single hairpin; at high concentrations, it co-migrates with the linear duplex d(A-G)10 x d(C-T)10, probably because it is a two-hairpin tetraplex. Molecular weights measured by equilibrium sedimentation in 0.1 M Na+, pH 7, reveal a mixture of monomer and dimer species at 1 degree C, but only a monomer at 40 degrees C; in 0.6 M Na+, pH 7, only a dimer species is observed at 4 degrees C. That the single- and double-stranded species are hairpin helices, is indicated by preferential S1 nuclease cleavage at the center of the oligomer(s), i.e., the loop of the hairpin(s). The UV melting transition below 0.3 M Na+ or K+, exhibits a dTm/dlog[Na+/K+] of 33 or 36 degrees C, respectively, consistent with conversion of a two-hairpin tetraplex to a single-hairpin duplex with extrahelical residues. When [Na+/K+] > or = 0.3 M, dTm/dlog [Na+/K+] is 19 or 17 degrees C, respectively, consistent with conversion of a two-hairpin tetraplex directly to single strands. A two-hairpin structure stabilized by G-tetrads is indicated by differential scanning calorimetry in 0.15 M Na+/5 mM Mg2+, with deltaH of formation per mole of the two-hairpin tetraplex of -116.9 kcal or -29.2 kcal/mol of G-tetrad. PMID:9016673

  19. DAG/PKCδ and IP3/Ca2+/CaMK IIβ Operate in Parallel to Each Other in PLCγ1-Driven Cell Proliferation and Migration of Human Gastric Adenocarcinoma Cells, through Akt/mTOR/S6 Pathway

    PubMed Central

    Dai, Lianzhi; Zhuang, Luhua; Zhang, Bingchang; Wang, Fen; Chen, Xiaolei; Xia, Chun; Zhang, Bing

    2015-01-01

    Phosphoinositide specific phospholipase Cγ (PLCγ) activates diacylglycerol (DAG)/protein kinase C (PKC) and inositol 1,4,5-trisphosphate (IP3)/Ca2+/calmodulin-dependent protein kinase II (CaMK II) axes to regulate import events in some cancer cells, including gastric adenocarcinoma cells. However, whether DAG/PKCδ and IP3/Ca2+/CaMK IIβ axes are simultaneously involved in PLCγ1-driven cell proliferation and migration of human gastric adenocarcinoma cells and the underlying mechanism are not elucidated. Here, we investigated the role of DAG/PKCδ or CaMK IIβ in PLCγ1-driven cell proliferation and migration of human gastric adenocarcinoma cells, using the BGC-823 cell line. The results indicated that the inhibition of PKCδ and CaMK IIβ could block cell proliferation and migration of BGC-823 cells as well as the effect of inhibiting PLCγ1, including the decrease of cell viability, the increase of apoptotic index, the down-regulation of matrix metalloproteinase (MMP) 9 expression level, and the decrease of cell migration rate. Both DAG/PKCδ and CaMK IIβ triggered protein kinase B (Akt)/mammalian target of rapamycin (mTOR)/S6 pathway to regulate protein synthesis. The data indicate that DAG/PKCδ and IP3/Ca2+/CaMK IIβ operate in parallel to each other in PLCγ1-driven cell proliferation and migration of human gastric adenocarcinoma cells through Akt/mTOR/S6 pathway, with important implication for validating PLCγ1 as a molecular biomarker in early gastric cancer diagnosis and disease surveillance. PMID:26633375

  20. InSAR analysis of the 2000 - 2002 earthquake sequence along the Sultan Dag front in the Isparta Angle (southern Turkey)

    NASA Astrophysics Data System (ADS)

    Manjunath, D. V.; Gomez, F. G.; Brooks, B. A.

    2009-12-01

    The Sultan Dag range of southern Turkey is a fault-block mountain range that has developed within or at the edge of the Isparta Angle - a region of the Anatolian plate situated near a tear in the subducting slab of the African plate. The mountain range is bounded on the northeast by the Sultandagi-Aksehir fault and the Aksehir-Afyon graben. A sequence of moderate earthquakes that occurred between 2000 and 2002 provides insight into the incremental growth of the mountain front through individual seismic events. The earthquake sequence consists of three moderate size events: a Mw 5.1 and a Mw 6.0 earthquake (December 15, 2000) and a Mw 6.5 earthquake (February 3, 2002). Surface deformations corresponding with the individual earthquakes were imaged using Interferometric Synthetic Aperture Radar (InSAR). The resulting displacement maps were used with elastic dislocation models to estimate faulting parameters associated with each earthquake (fault orientation and slip). These fault models were subsequently used to estimate static coulomb stress changes resulting from the earthquakes. The findings suggest that the two events of December 15, 2000, were not directly related to one another in terms of stress triggering. However, both events from 2000 appear to influence the extent of the 2002 earthquake. The larger event increased the stress on the segment ruptured in 2002, whereas the stress shadow from smaller event, which occurred on an antithetic fault, appears to have unloaded the stress at the termination of the 2002 rupture. Furthermore, the uplift and subsidence patterns associated with the earthquakes closely mimic overall geologic structure and general topography. Hence, these results permit exploring the extrapolation of coseismic displacements to understand the long-term growth of the mountain front.

  1. The role of PIP2 and the IP3/DAG pathway in intracellular calcium release and cell survival during nanosecond electric pulse exposures

    NASA Astrophysics Data System (ADS)

    Steelman, Zachary A.; Tolstykh, Gleb P.; Estlack, Larry E.; Roth, Caleb C.; Ibey, Bennett L.

    2015-03-01

    Phosphatidylinositol4,5-biphosphate (PIP2) is a membrane phospholipid of particular importance in cell-signaling pathways. Hydrolysis of PIP2 releases inositol-1,4,5-triphosphate (IP3) from the membrane, activating IP3 receptors on the smooth endoplasmic reticulum (ER) and facilitating a release of intracellular calcium stores and activation of protein kinase C (PKC). Recent studies suggest that nanosecond pulsed electric fields (nsPEF) cause depletion of PIP2 in the cellular membrane, activating the IP3 signaling pathway. However, the exact mechanism(s) causing this observed depletion of PIP2 are unknown. Complicating the matter, nsPEF create nanopores in the plasma membrane, allowing calcium to enter the cell and thus causing an increase in intracellular calcium. While elevated intracellular calcium can cause activation of phospholipase C (PLC) (a known catalyst of PIP2 hydrolysis), PIP2 depletion has been shown to occur in the absence of both extracellular and intracellular calcium. These observations have led to the hypothesis that the high electric field itself may be playing a direct role in the hydrolysis of PIP2 from the plasma membrane. To support this hypothesis, we used edelfosine to block PLC and prevent activation of the IP3/DAG pathway in Chinese Hamster Ovarian (CHO) cells prior to applying nsPEF. Fluorescence microscopy was used to monitor intracellular calcium bursts during nsPEF, while MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) survivability assays were utilized to determine whether edelfosine improved cell survival during nsPEF exposure. This work is critical to refine the role of PIP2 in the cellular response to nsPEF, and also to determine the fundamental biological effects of high electric field exposures.

  2. Evidence for the conformational rigidity of triplex d(C +T) 8-d(AG) 8·d(CT) 8 on silver electrode revealed by Fourier transform Raman scattering studies

    NASA Astrophysics Data System (ADS)

    Fang, Ye; Bai, Chunli; Wang, Ting; Zhong, Faping; Tang, Youqi; Lin, S. B.; Kan, Lou-sing

    1996-03-01

    Fourier transform surface enhanced Raman scattering spectroscopy (FT-SERS) has been first applied to characterize the triple stranded helix d(C +T) 8-d(AG) 8·d(CT) 8 (pH 4.5) (triplex) and its corresponding double-stranded helix d(AG) 8·d(CT) 8 (pH 7.0) (duplex) at an ex situ roughened silver electrode polarized at between 0.0 and -1.0 V vs. {Ag}/{AgCl}. The triplex adsorbed on the silver electrode yields five intense and one weak SERS bands located at 241, 839, 1189, 1293, 1643, and 1530 cm -1, respectively. These bands are not seen in the FT-SERS spectrum of the duplex, which showed a pattern similar to that of an oligo-DNA duplex observed by Koglin and Sequaries (Top. Curr. Chem., 134 (1989) 1). In the case of the triplex, the occurrence of the 839 cm -1 band with concomitant disappearance of the adsorbed adenine ring-breathing band at 734 cm -1 of the duplex indicates that the helical structure of the triplex near the surface of the silver electrode is well preserved and exhibits conformational rigidity. We observed a desorption process of the triplex from the silver electrode when the electrode approached -0.9 V (potential of zero charge of the triplex) vs. {Ag}/{AgCl}. This indicates that the highly negatively charged triplex shows a more favorable absorption on a highly positively charged silver surface. The structure of the triplex has also been studied by Fourier transform infrared (FTIR) and Raman spectroscopy. The vibrational spectra obtained clearly revealed that the conformation of the sugar moiety of the purine strand and one pyrimidine strand in the triplex is C 2'-endo/anti, whereas that of the other pyrimidine strand is C 3'-endo/anti.

  3. Transcription by eucaryotic and procaryotic RNA polymerases of DNA modified at a d(GG) or a d(AG) site by the antitumor drug cis-diamminedichloroplatinum (II)

    SciTech Connect

    Corda, Y.; Job, C.; Job, D. ); Anin, M-F.; Leng, M. )

    1991-01-01

    The authors have investigated whether DNA modifed at a d(GG) or a d(AG) site by the chemotherapeutic drug cis-diamminedichloroplatinum(II) (cis-DDP) can be used as template by wheat germ RNA polymerase II. The templates used in the present study were obtained by ligation of double-helical oligodeoxyribonucleotides, containing 18 pyrimidine bases and 2 central dG, or dA and dG, bases on one strand and 18 purine bases and 2 central dC, or dT and dC, bases on the complementary strand. Therefore, the cis-DDP adducts are only present on one strand of each of the two templates and are regularly spaced by 18 pyrimidine bases. These constructs allowed the authors to investigate the effect of cis-DDP on transcription of the platinated strand and of the complementary unplatinated sequence. Transcription experiments were carried out in the presence of dinucleotide primers and either a single triphosphate substrate (abortive elongation) or the full set of triphosphate substrates dictated by the template sequence (productive elongation). The results show taht the eucraryotic RNA polymerase can catalyze dinucleotide-primed reactions on platinated DNA. However, the eurcaryotic enzyme behaved very differently depending on which strand was transcribed. Thus, transcription elongation was completely blocked on the strand carrying the metal complex, whereas transcription elongation was not blocked on the complementary template strand. Similar results were obtained in transcription of the platinated templates by Escherichia coli RNA polymerase, suggesting that the cis-DDP-induced effect is associated with modifications of the basic catalytic properties of the transcriptases.

  4. Fast Reachability Test on DAGs for XML

    NASA Astrophysics Data System (ADS)

    Nakamura, Yusaku; Maita, Tetsuya; Sakamoto, Hiroshi

    We propose an efficient algorithm for deciding the reachability between any nodes on XML data represented by connected directed graphs. We develop a technique to reduce the size of the reference table for the reachability test. Using the small table and the standard range labeling method for rooted ordered trees, we show that our algorithm answers almost queries in a constant time preserving the space efficiency and a reasonable preprocessing time.

  5. Dynamic multi DAG scheduling algorithm for optical grid environment

    NASA Astrophysics Data System (ADS)

    Zhu, Liying; Sun, Zhenyu; Guo, Wei; Jin, Yaohui; Sun, Weiqiang; Hu, Weisheng

    2007-11-01

    Facing the evolvement of the Optical Grid technology, dynamic task scheduling can largely improve the efficiency of the Grid environment under the real circumstances. We propose a Serve On Time (SOT) algorithm - based on the idea of combining all the dynamic multi tasks so that all the tasks will obtain the rights to be served as soon as possible. We then introduce the basic First Come First Serve (FCFS) algorithm. A simulation will show the advantage of SOT.

  6. DAG Software Architectures for Multi-Scale Multi-Physics Problems at Petascale and Beyond

    NASA Astrophysics Data System (ADS)

    Berzins, Martin

    2015-03-01

    The challenge of computations at Petascale and beyond is to ensure how to make possible efficient calculations on possibly hundreds of thousands for cores or on large numbers of GPUs or Intel Xeon Phis. An important methodology for achieving this is at present thought to be that of asynchronous task-based parallelism. The success of this approach will be demonstrated using the Uintah software framework for the solution of coupled fluid-structure interaction problems with chemical reactions. The layered approach of this software makes it possible for the user to specify the physical problems without parallel code, for that specification to be translated into a parallel set of tasks. These tasks are executed using a runtime system that executes tasks asynchronously and sometimes out-of-order. The scalability and portability of this approach will be demonstrated using examples from large scale combustion problems, industrial detonations and multi-scale, multi-physics models. The challenges of scaling such calculations to the next generations of leadership class computers (with more than a hundred petaflops) will be discussed. Thanks to NSF, XSEDE, DOE NNSA, DOE NETL, DOE ALCC and DOE INCITE.

  7. New DAG and cAMP Sensors Optimized for Live-Cell Assays in Automated Laboratories

    PubMed Central

    Tewson, Paul H.; Martinka, Scott; Shaner, Nathan C.; Hughes, Thomas E.; Quinn, Anne Marie

    2015-01-01

    Protein-based, fluorescent biosensors power basic research on cell signaling in health and disease, but their use in automated laboratories is limited. We have now created two live-cell assays, one for diacyl glycerol and another for cAMP, that are robust (Z′ > 0.7) and easily deployed on standard fluorescence plate readers. We describe the development of these assays, focusing on the parameters that were critical for optimization, in the hopes that the lessons learned can be generalized to the development of new biosensor-based assays. PMID:26657040

  8. Identification of TAG and DAG and their FA constituents in Lesquerella (Physaria fendleri) oil by HPLC and MS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Castor oil has many industrial uses because of its high content (90 %) of the hydroxy fatty acid, ricinoleic acid (OH1218:19). Lesquerella oil containing lesquerolic acid (Ls, OH1420:111) is potentially useful in industry. Ten molecular species of diacylglycerols and 74 molecular species of triacylg...

  9. Vitellogenesis in the fruit fly, Drosophila melanogaster: antagonists demonstrate that the PLC, IP3/DAG, PK-C pathway is triggered by calmodulin.

    PubMed

    Brubaker-Purkey, Bethany J; Woodruff, Richard I

    2013-01-01

    In Drosophila melanogaster M. (Diptera: Drosophilidae), a phospholipase-C to proteininase-C signal cascade leads to the endocytic uptake of yolk precursor molecules. The data suggest that D. melanogaster has a phospholipase-C/proteinkinase-C signaling pathway similar to that previously shown to be required for vitellogenesis in the milkweed bug, Oncopeltus fasciatus Dallas (Hemiptera: Lygaeidae). Calmodulin, derived from epithelial cells and transported to the oocytes via gap junctions, may trigger this pathway. To investigate this, a series of known antagonists to various elements of the pathway were used. W-7 (which prevents calmodulin binding to phospholipase-C), U-73122 (which prevents activation of phospholipase-C), verapamil (which blocks Ca(2+) release by IP3), HAG (which blocks diacylglycerol), and staurosporine (which inactivates proteinkinase-C) were each shown to inhibit endocytosis, thereby blocking formation of nascent yolk spheres. PMID:24228869

  10. Vitellogenesis in the Fruit Fly, Drosophila melanogaster: Antagonists Demonstrate that the PLC, IP3/DAG, PK-C Pathway is Triggered by Calmodulin

    PubMed Central

    Brubaker-Purkey, Bethany J.; Woodruff, Richard I.

    2013-01-01

    In Drosophila melanogaster M. (Diptera: Drosophilidae), a phospholipase-C to proteininase-C signal cascade leads to the endocytic uptake of yolk precursor molecules. The data suggest that D. melanogaster has a phospholipase-C/proteinkinase-C signaling pathway similar to that previously shown to be required for vitellogenesis in the milkweed bug, Oncopeltus fasciatus Dallas (Hemiptera: Lygaeidae). Calmodulin, derived from epithelial cells and transported to the oocytes via gap junctions, may trigger this pathway. To investigate this, a series of known antagonists to various elements of the pathway were used. W-7 (which prevents calmodulin binding to phospholipase-C), U-73122 (which prevents activation of phospholipase-C), verapamil (which blocks Ca2+ release by IP3), HAG (which blocks diacylglycerol), and staurosporine (which inactivates proteinkinase-C) were each shown to inhibit endocytosis, thereby blocking formation of nascent yolk spheres. PMID:24228869

  11. 28 CFR 16.71 - Exemption of the Office of the Deputy Attorney General System-limited access.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... Attorney General System-limited access. 16.71 Section 16.71 Judicial Administration DEPARTMENT OF JUSTICE... Records System (JUSTICE/DAG-006). (2) Presidential Appointee Records System (JUSTICE/DAG-007). (3) Special Candidates for Presidential Appointments Records System (JUSTICE/DAG-008). (4) Miscellaneous...

  12. 28 CFR 16.71 - Exemption of the Office of the Deputy Attorney General System-limited access.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... Attorney General System-limited access. 16.71 Section 16.71 Judicial Administration DEPARTMENT OF JUSTICE... Records System (JUSTICE/DAG-006). (2) Presidential Appointee Records System (JUSTICE/DAG-007). (3) Special Candidates for Presidential Appointments Records System (JUSTICE/DAG-008). (4) Miscellaneous...

  13. 28 CFR 16.71 - Exemption of the Office of the Deputy Attorney General System-limited access.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... Attorney General System-limited access. 16.71 Section 16.71 Judicial Administration DEPARTMENT OF JUSTICE... Records System (JUSTICE/DAG-006). (2) Presidential Appointee Records System (JUSTICE/DAG-007). (3) Special Candidates for Presidential Appointments Records System (JUSTICE/DAG-008). (4) Miscellaneous...

  14. 28 CFR 16.71 - Exemption of the Office of the Deputy Attorney General System-limited access.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... Attorney General System-limited access. 16.71 Section 16.71 Judicial Administration DEPARTMENT OF JUSTICE... Records System (JUSTICE/DAG-006). (2) Presidential Appointee Records System (JUSTICE/DAG-007). (3) Special Candidates for Presidential Appointments Records System (JUSTICE/DAG-008). (4) Miscellaneous...

  15. 28 CFR 16.71 - Exemption of the Office of the Deputy Attorney General System-limited access.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... Attorney General System-limited access. 16.71 Section 16.71 Judicial Administration DEPARTMENT OF JUSTICE... Records System (JUSTICE/DAG-006). (2) Presidential Appointee Records System (JUSTICE/DAG-007). (3) Special Candidates for Presidential Appointments Records System (JUSTICE/DAG-008). (4) Miscellaneous...

  16. Acyl migration kinetics of vegetable oil 1,2-diacylglycerols

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The acyl migration kinetics of long-chain 1,2-diacylglycerol (1,2-DAG) to form 1,3-diacylglycerol (1,3-DAG) over the temperature range of 25 to 80 degrees Celsius were examined using proton NMR spectroscopy. The 1,2-DAG mole fraction of 0.32 at equilibrium was found to be insensitive to temperature...

  17. Total body water and fat-free mass: evaluation of equations based on bioelectrical impedance analysis in infants and young children in India.

    PubMed

    Sen, Bandana; Mahalanabis, Dilip; Kurpad, Anura V; Shaikh, Saijuddin; Bose, Kaushik

    2010-07-01

    The association of early postnatal growth with diseases in adults such as hypertension, type 2 diabetes and CHD has generated interest in studying postnatal growth. Bioelectrical impedance analysis (BIA) is a useful measure to estimate total body water (TBW) and fat-free mass (FFM). We evaluated three published equations (Fjeld et al. (Pediatr Res (1990) 27, 98-102), Bocage (MSc Thesis (1988) University of West Indies) and Kushner et al. (Am J Clin Nutr (1992) 56, 835-839) to measure TBW and derived FFM based on BIA, using 2H2O dilution as a reference method for suitability in infants in India. In a cross-sectional study in seventy-eight apparently healthy infants aged 6-24 months from the urban poor attending an immunisation clinic at a hospital in Kolkata, we measured their length to the nearest 0.1 cm, weight to the nearest 10 g, resistance at 50 kHz using BIA and TBW using 2H2O dilution. TBW was derived using three published BIA-based equations and compared with TBW using 2H2O dilution. Based on the BIA equations of Fjeld et al., Bocage and Kushner et al., the mean TBW values were 2.46 % (P < 0.001), 4.62 % (P < 0.001) and 9.50 % (P < 0.001) lower than the reference 2H2O method, respectively. All three published BIA-based equations consistently underestimated the TBW and FFM and appeared inadequate for studying infants in India. The equation described by Fjeld et al. gave the smallest deviation from the reference method and may be used for field studies. New equations based on population-specific data are desirable for a more precise measure of TBW. PMID:20188001

  18. Alteration of lipid membrane structure and dynamics by diacylglycerols with unsaturated chains.

    PubMed

    Alwarawrah, Mohammad; Hussain, Fazle; Huang, Juyang

    2016-02-01

    Diacylglycerols (DAGs) with unsaturated acyl chains play many important roles in biomembranes, such as a second messenger and activator for protein kinase C. In this study, three DAGs of distinctly different chain unsaturations (i.e. di16:0DAG (DPG), 16:0-18:1DAG (POG), and di18:1DAG (DOG)) are studied using atomistic MD simulation to compare their roles in the structure and dynamics of 16:0-18:1phosphatidylcholine (POPC) membranes. All three DAGs are able to produce the so-called 'condensing effect' in POPC membranes: decreasing area-per-lipid, and increasing acyl chain order and bilayer thickness. Our visual and quantitative analyses clearly show that DAG with unsaturated chains induce larger spacing between POPC headgroups, compared with DAG with saturated chains; this particular effect has long been hypothesized to be crucial for activating enzymes and receptors in cell membranes. DAGs with unsaturated chains are also located closer to the bilayer/aqueous interface than DPG and are more effective in slowing down lateral diffusion of molecules. We show that DAG molecules seek the "umbrella coverage" from neighboring phospholipid headgroups - similar to cholesterol. Unlike cholesterol, DAGs also hide their chains from water by laterally inserting their chains into the surrounding. Thus, acyl chains of DAG are more spread and disordered than those of PC due to the insertion. By calculating the potential of mean force (PMF) for POPC in POPC/DAG bilayers, we found that all three DAGs can significantly increase the free energy barrier for POPC to flip-flop, but only DAGs with unsaturated chains can additionally increase the free energy of POPC desorption. PMID:26607007

  19. Sphingomyelin regulates the transbilayer movement of diacylglycerol in the plasma membrane of Madin-Darby canine kidney cells.

    PubMed

    Ueda, Yoshibumi; Makino, Asami; Murase-Tamada, Kotono; Sakai, Shota; Inaba, Takehiko; Hullin-Matsuda, Françoise; Kobayashi, Toshihide

    2013-08-01

    Diacylglycerol (DAG) is a key component in lipid metabolism and signaling. Previous model membrane studies using DAG analogs suggest their rapid membrane transbilayer movement. However, little is known about the DAG distribution and dynamics in cell membranes. Using live-cell fluorescence microscopy, we monitored the transbilayer movement of DAG with the yellow fluorescent protein-tagged C1AB domain from protein kinase C-γ (EYFP-C1AB), which selectively binds DAG. When HeLa cells were treated with Bacillus cereus phospholipase C (Bc-PLC) to produce DAG on the outer leaflet of the plasma membrane, intracellularly expressed EYFP-C1AB probe accumulated at the plasma membrane, indicating the transbilayer movement of the outer leaflet DAG to the inner leaflet. This Bc-PLC-induced translocation of EYFP-C1AB probe to the plasma membrane was not observed in the sphingolipid-enriched plasma membrane of Madin-Darby canine kidney cells, but was recovered after cell treatment with sphingomyelinase or preincubation with an inhibitor of sphingolipid biosynthesis. The inhibitory effect of sphingomyelin (SM) on the transbilayer movement of DAG was reproduced in model membranes using a fluorescent short-chain DAG analog. These results demonstrate that the SM content on the outer leaflet regulates the transbilayer movement of DAG in the plasma membrane, thus providing new insights into the dynamics of DAG in cell pathophysiology. PMID:23682124

  20. Photoswitchable diacylglycerols enable optical control of protein kinase C.

    PubMed

    Frank, James Allen; Yushchenko, Dmytro A; Hodson, David J; Lipstein, Noa; Nagpal, Jatin; Rutter, Guy A; Rhee, Jeong-Seop; Gottschalk, Alexander; Brose, Nils; Schultz, Carsten; Trauner, Dirk

    2016-09-01

    Increased levels of the second messenger lipid diacylglycerol (DAG) induce downstream signaling events including the translocation of C1-domain-containing proteins toward the plasma membrane. Here, we introduce three light-sensitive DAGs, termed PhoDAGs, which feature a photoswitchable acyl chain. The PhoDAGs are inactive in the dark and promote the translocation of proteins that feature C1 domains toward the plasma membrane upon a flash of UV-A light. This effect is quickly reversed after the termination of photostimulation or by irradiation with blue light, permitting the generation of oscillation patterns. Both protein kinase C and Munc13 can thus be put under optical control. PhoDAGs control vesicle release in excitable cells, such as mouse pancreatic islets and hippocampal neurons, and modulate synaptic transmission in Caenorhabditis elegans. As such, the PhoDAGs afford an unprecedented degree of spatiotemporal control and are broadly applicable tools to study DAG signaling. PMID:27454932

  1. A cladistically based reinterpretation of the taxonomy of two Afrotropical tenebrionid genera Ectateus Koch, 1956 and Selinus Mulsant & Rey, 1853 (Coleoptera, Tenebrionidae, Platynotina)

    PubMed Central

    Kamiński, Marcin Jan

    2014-01-01

    Abstract On the basis of a newly performed cladistic analysis a new classification of the representatives of two Afrotropical tenebrionid genera, Ectateus Koch, 1956 and Selinus Mulsant & Rey, 1853 sensu Iwan 2002a, is provided. Eleoselinus is described as a new genus. The genus Monodius, previously synonymized with Selinus by Iwan (2002), is redescribed and considered as a separate genus. Following new combinations are proposed: Ectateus calcaripes (Gebien, 1904), Monodius laevistriatus (Fairmaire, 1897), Monodius lamottei (Gridelli, 1954), Monodius plicicollis (Fairmaire, 1897), Eleoselinus villiersi (Ardoin, 1965) and Eleoselinus ursynowiensis (Kamiński, 2011). Neotype for Ectateus calcaripes and lectotypes for E. crenatus (Fairmaire, 1897), E. ghesquierei Koch, 1956 and Monodius malaisei malaisei Koch, 1956 are designated to fix the taxonomic status of these taxa. The following synonymies are proposed: Selinus monardi Kaszab, 1951 and Ectateus latipennis Koch, 1956 with E. crenatus (Fairmaire, 1897). Identification keys are provided to all known species of Ectateus sensu novum, Eleoselinus, Monodius and Selinus sensu novum. PMID:25009425

  2. A cladistically based reinterpretation of the taxonomy of two Afrotropical tenebrionid genera Ectateus Koch, 1956 and Selinus Mulsant & Rey, 1853 (Coleoptera, Tenebrionidae, Platynotina).

    PubMed

    Kamiński, Marcin Jan

    2014-01-01

    On the basis of a newly performed cladistic analysis a new classification of the representatives of two Afrotropical tenebrionid genera, Ectateus Koch, 1956 and Selinus Mulsant & Rey, 1853 sensu Iwan 2002a, is provided. Eleoselinus is described as a new genus. The genus Monodius, previously synonymized with Selinus by Iwan (2002), is redescribed and considered as a separate genus. Following new combinations are proposed: Ectateus calcaripes (Gebien, 1904), Monodius laevistriatus (Fairmaire, 1897), Monodius lamottei (Gridelli, 1954), Monodius plicicollis (Fairmaire, 1897), Eleoselinus villiersi (Ardoin, 1965) and Eleoselinus ursynowiensis (Kamiński, 2011). Neotype for Ectateus calcaripes and lectotypes for E. crenatus (Fairmaire, 1897), E. ghesquierei Koch, 1956 and Monodius malaisei malaisei Koch, 1956 are designated to fix the taxonomic status of these taxa. The following synonymies are proposed: Selinus monardi Kaszab, 1951 and Ectateus latipennis Koch, 1956 with E. crenatus (Fairmaire, 1897). Identification keys are provided to all known species of Ectateus sensu novum, Eleoselinus, Monodius and Selinus sensu novum. PMID:25009425

  3. The Significance of Different Diacylgycerol Synthesis Pathways on Plant Oil Composition and Bioengineering

    PubMed Central

    Bates, Philip D.; Browse, John

    2012-01-01

    The unique properties of vegetable oils from different plants utilized for food, industrial feedstocks, and fuel is dependent on the fatty acid (FA) composition of triacylglycerol (TAG). Plants can use two main pathways to produce diacylglycerol (DAG), the immediate precursor molecule to TAG synthesis: (1) De novo DAG synthesis, and (2) conversion of the membrane lipid phosphatidylcholine (PC) to DAG. The FA esterified to PC are also the substrate for FA modification (e.g., desaturation, hydroxylation, etc.), such that the FA composition of PC-derived DAG can be substantially different than that of de novo DAG. Since DAG provides two of the three FA in TAG, the relative flux of TAG synthesis from de novo DAG or PC-derived DAG can greatly affect the final oil FA composition. Here we review how the fluxes through these two alternate pathways of DAG/TAG synthesis are determined and present evidence that suggests which pathway is utilized in different plants. Additionally, we present examples of how the endogenous DAG synthesis pathway in a transgenic host plant can produce bottlenecks for engineering of plant oil FA composition, and discuss alternative strategies to overcome these bottlenecks to produce crop plants with designer vegetable oil compositions. PMID:22783267

  4. Production of diacylglycerols from glycerol monooleate and ethyl oleate through free and immobilized lipase-catalyzed consecutive reactions.

    PubMed

    Jin, Juan; Li, Dan; Zhu, Xue Mei; Adhikari, Prakash; Lee, Ki-Teak; Lee, Jeung-Hee

    2011-02-28

    The ability of free and immobilized lipase on the production of diacylglycerols (DAG) by transesterification of glycerol monooleate (GMO) and ethyl oleate was investigated. Among three free lipases such as lipase G (Penicillium cyclopium), lipase AK (Pseudomonas fluorescens) and lipase PS (Pseudomonas cepacia), lipase PS exhibited the highest DAG productivity, and the DAG content gradually increased up to 24 hours reaction and then remained steady. The comparative result for DAG productivity between free lipase PS and immobilized lipases (lipase PS-D and Lipozyme RM IM) during nine times of 24 hours reaction indicated that total DAG production was higher in immobilized lipase PS-D (183.5mM) and Lipozyme RM IM (309.5mM) than free lipase PS (122.0mM) at the first reaction, and that the DAG production rate was reduced by consecutive reactions, in which more sn-1,3-DAG was synthesized than sn-1,2-DAG. During the consecutive reactions, the activity of lipase PS was relatively steady by showing similar DAG content, whereas DAG production of lipase PS-D and Lipozyme RM IM was gradually decreased to 69.9 and 167.1mM at 9th reaction, respectively, resulting in 62% and 46% reduced production when compared with 1st reaction. Interestingly, from 7th reaction lipase PS produced more DAG than immobilized lipase PS-D, and exhibited a stable activity for DAG production. Therefore, the present study suggested that DAG productivity between GMO and ethyl oleate was higher in immobilized lipases than free lipases, but the activity was reduced with repeated uses. PMID:20951847

  5. Identification of molecular species of simple lipids by normal phase liquid chromatography-positive electrospray tandem mass spectrometry, and application of developed methods in comprehensive analysis of low erucic acid rapeseed oil lipids

    NASA Astrophysics Data System (ADS)

    Kalo, P. J.; Ollilainen, V.; Rocha, J. M.; Malcata, F. X.

    2006-07-01

    Mono-, di- and triacylglycerol (MAG, DAG, TAG), sterol ester (SE), free sterol (S) and free fatty acid (FFA) standards were analyzed in the presence of ammonium ions and ammonia by flow injection MS2 and MS3, and by normal phase-liquid chromatography (NP-LC) MS2 positive electrospray ionization (ESI) mass spectrometry (MS). The MS data recorded for ammonium adducts ([M + NH4]+) of TAGs, DAGs, and MAGs were consistent with stepwise fragmentation mechanisms. In the first step, ammonium ion in [M + NH4]+ donates proton to acylglycerol and ammonia is released. In the second step, FFA is cleaved from protonated TAG, water from protonated 1,3-DAG and MAG, both FFA and water from protonated 1,2-DAG, hence leading to formation of [DAG]+ ion from TAG and 1,3-DAG, [DAG]+ and [MAG]+ ions from 1,2-DAG, and [MAG]+ ion from MAG. In the third step, [DAG]+ ion of TAG is fragmented to yield [Acyl]+, [Acyl + 74]+, [DAG - 74]+ ions, [DAG] ion of 1,3-DAG to [Acyl]+ ions, and [MAG]+ ion of MAG to protonated FAs, which are decomposed to water and [Acyl]+ ions in the fourth step. A stepwise mechanism for fragmentation of FFA was also evident from MS2 and MS3 data. Molecular species of low erucic acid rapeseed oil simple lipids were identified from characteristic ions produced in the NP-LC-ESI-MS2 of [M + NH4]+ ions. The percentage composition of the molecular species of each lipid class was calculated from integrated extracted ion chromatograms of [(M + NH4)]+ ions of SE, TAG, MAG, and FFA, of the sum of [(M + NH4)]+ and [(M + NH4) - NH3 - H2O]+ ions of both regioisomers of DAGs, and of sterol fragment ions of S.

  6. Spatiotemporal dynamics of lipid signaling: protein kinase C as a paradigm

    PubMed Central

    Gallegos, Lisa L.; Newton, Alexandra C.

    2009-01-01

    Summary The lipid second messenger diacylglycerol (DAG) acutely controls the rate, amplitude, duration, and location of protein kinase C (PKC) activity in the cell. There are three classes of PKC isozymes and, of these, the conventional and novel isozymes are acutely controlled by DAG. The kinetics of DAG production at various intracellular membranes, the intrinsic affinity of specific isoforms for DAG-containing membranes, the coordinated use of additional membrane-binding modules, the intramolecular regulation of DAG sensitivity, and the competition from other DAG-responsive proteins together result in a unique, context-dependent activation signature for each isoform. This review focuses on the spatiotemporal dynamics of PKC activation and how it is controlled by lipid second messengers. PMID:18720411

  7. Endothelin stimulates a sustained 1,2-diacylglycerol increase and protein kinase C activation in bovine aortic smooth muscle cells

    SciTech Connect

    Lee, T.S.; Chao, T.; Hu, K.Q.; King, G.L.

    1989-07-14

    Endothelin is a long-lasting potent vasoconstrictor peptide. We report here that in bovine aortic smooth muscle cells, endothelin biphasically increased total cellular diacylglycerol (DAG) content. When cellular DAG was labeled with (/sup 14/C) glycerol for 48h, endothelin stimulated (/sup 14/C)DAG formation in a biphasic pattern. Only one prolonged phase of DAG accumulation was observed when cells were labeled with (/sup 3/H)glycerol for 2 h. Endothelin induced an increase in the membranous protein kinase C (PKC) activities, which lasted for more than 20 min. These data suggest that (i) endothelin stimulates a sustained generation of DAG, (ii) this accumulation of DAG results in a sustained translocation of cytosolic PKC activities to the membrane.

  8. Regulation of the transbilayer movement of diacylglycerol in the plasma membrane.

    PubMed

    Ueda, Yoshibumi; Ishitsuka, Reiko; Hullin-Matsuda, Françoise; Kobayashi, Toshihide

    2014-12-01

    This mini-review presents recent advances in the regulation of the membrane transbilayer movement (or flip-flop) of diacylglycerol (DAG), a key intermediate in lipid metabolism and a second messenger in lipid-mediated signaling. Despite progresses in lipid biophysics and imaging, little is known about the DAG dynamics across the two leaflets of the plasma membrane in living cells. Previous model membrane studies with DAG analogs demonstrated their fast flip-flop suggesting that DAG is evenly distributed between the two leaflets of the plasma membrane. However, recent molecular dynamics simulations indicate that DAG transbilayer movement depends on the lipid environment surrounding the lipid, i.e. DAG flips more slowly across a more ordered "lipid raft-like" bilayer (enriched in sphingomyelin/cholesterol) than across a more fluid bilayer (composed of unsaturated glycerophospholipids). Furthermore using the yellow fluorescent protein-tagged C1AB domain from protein kinase C-γ (EYFP-C1AB) that selectively binds DAG, we recently proved that the sphingomyelin (SM) content in the plasma membrane outer leaflet regulates DAG transbilayer movement in Madin-Darby canine kidney cells treated with bacterial phosphatidylcholine-specific phospholipase C. The dose-dependent inhibition of DAG flip-flop by SM could be reproduced in model membranes using fluorescent short chain DAG analog. Regulation of DAG transbilayer movement by the outer leaflet SM content is expected to modify the downstream recruitment of C1-domain containing effectors, thus bringing new insights on the role of DAG dynamics in cell pathophysiology. PMID:25241257

  9. Characterization of diacylglycerol isomers in edible oils using gas chromatography-ion trap electron ionization mass spectrometry.

    PubMed

    Zhu, Hanjiang; Clegg, Michael S; Shoemaker, Charles F; Wang, Selina C

    2013-08-23

    Verifying the authenticity of edible oils is of international concern. A new quality control standard for olive oil has been proposed that relates the ratio of 1,2-diacylglycerol (DAG) to 1,3-DAG to sensory aspects of olive oil. DAGs and their isomers are difficult to quantitate and characterize by Flame Ionization Gas Chromatography (GC-FID) due to the lack of suitable standards. Mass detectors offer the advantage of providing structural detail to the eluding DAG(s), thus removing ambiguity to the identification of both resolved and unresolved DAGs in GC chromatograms. In this study, a GC Electron Ionization Mass Spectrometry (GC-EI-MS) method was developed to determine the fatty acid composition and molecular structure of trimethylsilyl (TMS) derivatized DAGs present in edible oils. Twenty-two species of DAG isomers were identified in refined coconut oil and unrefined olive oil utilizing signature fragment ions, [M-15](+), [M-89](+), [M-RCO2](+), [RCO2+58](+) and [M-RCO2CH2](+). The [M-RCO2CH2](+) ion is considered the key diagnostic ion to distinguish between DAG positional isomers. MS/MS spectra of [M-RCO2](+) and [M-15](+) ions obtained from commercial standards containing both 1,2- and 1,3-DAG isomers were used as a model system to confirm the identification of DAG isomers in natural products. Furthermore, a number of reaction mechanisms are proposed to explain the formation of the most abundant mass fragments of DAGs and their isomers. PMID:23880469

  10. Assay for CDP-Diacylglycerol Generation by CDS in Membrane Fractions.

    PubMed

    Waugh, Mark G

    2016-01-01

    CDP-DAG is a liponucleotide formed by the condensation of CTP with the phospholipid phosphatidic acid in a reaction catalyzed by CDP-DAG synthase (CDS). CDP-DAG is required for the synthesis of phosphatidylinositol; the parent molecule whence all seven phosphoinositides including the signaling molecules PI4P, PI(4,5)P2, and PI(3,4,5)P3 are derived. This protocol describes a highly sensitive radiometric assay to detect the generation of CDP-DAG on isolated biological membrane fractions. PMID:26552690

  11. Analysis of diacylglycerols by ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry: Double bond location and isomers separation.

    PubMed

    Deng, Pan; Zhong, Dafang; Wang, Xi; Dai, Yulu; Zhou, Lei; Leng, Ying; Chen, Xiaoyan

    2016-06-21

    Diacylglycerols (DAGs) are important lipid intermediates and have been implicated in human diseases. Isomerism complicates their mass spectrometric analysis; in particular, it is difficult to identify fatty acid substituents and locate the double bond positions in unsaturated DAGs. We have developed an analytical strategy using ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF MS) in conjunction with dimethyl disulfide (DMDS) derivatization and collision cross-section (CCS) measurement to characterize DAGs in biological samples. The method employs non-aqueous reversed-phase chromatographic separation and profile collision energy (CE) mode for MS(E) and MS/MS analyses. Three types of fragment ions were produced simultaneously. Hydrocarbon ions (m/z 50-200) obtained at high CE helped to distinguish unsaturated and saturated DAGs rapidly. Neutral loss ions and acylium ions (m/z 300-400) produced at low CE were used to identify fatty acid substituents. Informative methyl thioalkane fragment ions were used to locate the double bonds of unsaturated DAGs. Mono-methylthio derivatives were formed mainly by the reaction of DAGs with DMDS, where methyl thiol underwent addition to the first double bond farthest from the ester terminus of unsaturated fatty acid chains. The addition of CCS values maximized the separation of isomeric DAG species and improved the confidence of DAG identification. Fourteen DAGs were identified in mouse myotube cells based on accurate masses, characteristic fragment ions, DMDS derivatization, and CCS values. PMID:27188314

  12. Control of TRPC and store-operated channels by protein kinase C.

    PubMed

    Venkatachalam, Kartik; Zheng, Fei; Gill, Donald L

    2004-01-01

    TRPC channels are widely expressed among cells and are believed to play important roles in receptor-mediated Ca2+ signalling. We determined that the function of TRPC channels is highly regulated by protein kinase C (PKC). Application of diacylglycerol (DAG) or elevated endogenous DAG resulting from either DAG-lipase or DAG-kinase inhibition, completely prevented TRPC5 or TRPC4 activation in both HEK293 cells and DT40 cells. This inhibitory action of DAG on TRPC5 and TRPC4 channels was clearly mediated by PKC, in distinction to the stimulatory action of DAG on TRPC3 which was PKC-independent. PKC activation totally blocked TRPC3 channel-activated in response to OAG, and was restored by PKC-blockade. PKC-inhibition resulted in decreased TRPC3 channel deactivation. Store-operated Ca2+ entry in response to PLC-coupled receptor activation but not store-depletion per se, was substantially reduced by OAG or DAG-lipase inhibition in a PKC-dependent manner. The results reveal that each TRPC subtype is strongly inhibited by DAG-induced PKC activation reflecting a likely universal feedback control on TRPCs. The profound yet distinct control by PKC and DAG on the activation of TRPC channel subtypes may be the basis of a spectrum of regulatory phenotypes of expressed TRPC channels. PMID:15104182

  13. Identification of a Common R-Chondrite Impactor on the Ureilite Parent Body

    NASA Technical Reports Server (NTRS)

    Downes, H.; Mittlefehldt, D. W.

    2006-01-01

    Polymict ureilites are brecciated ultramafic meteorites that contain a variety of single mineral and lithic clasts. They represent the surface debris from a small, differentiated asteroid. We are continuing a detailed petrological study of several polymict ureilites including EET 87720, EET 83309 and FRO93008 (from Antarctica), North Haig, Nilpena (Australia), DaG 976, DaG 999, DaG 1000 and DaG 1023 (Libya). The latter four stones are probably paired. Clast sizes can be 10 mm in diameter, so a thin-section can consist of a single lithic clast.

  14. Des-Acyl Ghrelin Directly Targets the Arcuate Nucleus in a Ghrelin-Receptor Independent Manner and Impairs the Orexigenic Effect of Ghrelin.

    PubMed

    Fernandez, G; Cabral, A; Cornejo, M P; De Francesco, P N; Garcia-Romero, G; Reynaldo, M; Perello, M

    2016-02-01

    Ghrelin is a stomach-derived octanoylated peptide hormone that plays a variety of well-established biological roles acting via its specific receptor known as growth hormone secretagogue receptor (GHSR). In plasma, a des-octanoylated form of ghrelin, named des-acyl ghrelin (DAG), also exists. DAG is suggested to be a signalling molecule that has specific targets, including the brain, and regulates some physiological functions. However, no specific receptor for DAG has been reported until now, and, consequently, the potential role of DAG as a hormone has remained a matter of debate. In the present study, we show that DAG specifically binds to and acts on a subset of arcuate nucleus (ARC) cells in a GHSR-independent manner. ARC cells labelled by a DAG fluorescent tracer include the neuropeptide Y (NPY) and non-NPY neurones. Given the well-established role of the ARC in appetite regulation, we tested the effect of centrally administered DAG on food intake. We found that DAG failed to affect dark phase feeding, as well as food intake, after a starvation period; however, it impaired the orexigenic actions of peripherally administered ghrelin. Thus, we conclude that DAG directly targets ARC neurones and antagonises the orexigenic effects of peripherally administered ghrelin. PMID:26661382

  15. Identification and disruption of an Arabidopsis zinc finger gene controlling seed germination

    PubMed Central

    Papi, Maura; Sabatini, Sabrina; Bouchez, David; Camilleri, Christine; Costantino, Paolo; Vittorioso, Paola

    2000-01-01

    We describe here the Arabidopsis gene DAG1, encoding a zinc finger transcription factor of the Dof family, and show that it is involved in the control of seed germination. By a reverse genetics approach, we isolated an Arabidopsis mutant line with one T-DNA insertion in DAG1. Seeds from homozygous knockout dag1-1 plants do not develop dormancy and germinate also in the absence of light. Segregation analysis indicates that the effect of the mutation is maternal. Accordingly, in situ mRNA hybridizations reveal expression of DAG1 in the vascular tissue of the flower and maturing fruit but not in the seed. PMID:10640273

  16. Monoglucosyldiacylglycerol, a foreign lipid, can substitute for phosphatidylethanolamine in essential membrane-associated functions in Escherichia coli.

    PubMed

    Wikström, Malin; Xie, Jun; Bogdanov, Mikhail; Mileykovskaya, Eugenia; Heacock, Philip; Wieslander, Ake; Dowhan, William

    2004-03-12

    The mechanisms by which lipid bilayer properties govern or influence membrane protein functions are little understood, but a liquid-crystalline state and the presence of anionic and nonbilayer (NB)-prone lipids seem important. An Escherichia coli mutant lacking the major membrane lipid phosphatidylethanolamine (NB-prone) requires divalent cations for viability and cell integrity and is impaired in several membrane functions that are corrected by introduction of the "foreign" NB-prone neutral glycolipid alpha-monoglucosyldiacylglycerol (MGlcDAG) synthesized by the MGlcDAG synthase from Acholeplasma laidlawii. Dependence on Mg(2+) was reduced, and cellular yields and division malfunction were greatly improved. The increased passive membrane permeability of the mutant was not abolished, but protein-mediated osmotic stress adaptation to salts and sucrose was recovered by the presence of MGlcDAG. MGlcDAG also restored tryptophan prototrophy and active transport function of lactose permease, both critically dependent on phosphatidylethanolamine. Three mechanisms can explain the observed effects: NB-prone MGlcDAG improves the quenched lateral pressure profile across the bilayer; neutral MGlcDAG dilutes the high anionic lipid surface charge; MGlcDAG provides a neutral lipid that can hydrogen bond and/or partially ionize. The reduced dependence on Mg(2+) and lack of correction by high monovalent salts strongly support the essential nature of the NB properties of MGlcDAG. PMID:14688287

  17. Purification of 1,2-diacylglycerols from vegetable oils: comparison of molecular distillation and liquid CO2 extraction

    Technology Transfer Automated Retrieval System (TEKTRAN)

    High oleic sunflower oil and soybean oil were partially deacylated by enzyme-catalyzed propanolysis using Lipozyme TL IM to form 1,2-diacyl-sn-glycerols (1,2-DAG) in 40% yields. 1,2-DAG are of interest as potential regiospecific intermediates in the synthesis of structured lipids for cosmeceutical ...

  18. The Abundance and Isotopic Composition of Water in Howardite-Eucrite-Diogenite Meteorites

    NASA Astrophysics Data System (ADS)

    Barrett, T. J.; Tartèse, R.; Anand, M.; Franchi, I. A.; Grady, M. M.; Greenwood, R. C.; Charlier, B. L. A.

    2014-09-01

    Using SIMs techniques we measure OH abundances and D/H ratios in apatite grains from two Eucrites (DaG 945, DaG 844).The average δD values of these two samples are also similar to carbonaceous chondrites, the Earth and the Moon.

  19. Alpha Linolenic Acid-enriched Diacylglycerol Enhances Postprandial Fat Oxidation in Healthy Subjects: A Randomized Double-blind Controlled Trail.

    PubMed

    Ando, Yasutoshi; Saito, Shinichiro; Oishi, Sachiko; Yamanaka, Nami; Hibi, Masanobu; Osaki, Noriko; Katsuragi, Yoshihisa

    2016-08-01

    Alpha linolenic acid-enriched diacylglycerol (ALA-DAG) reduces visceral fat area and body fat in rodents and humans compared to conventional triacylglycerol (TAG). Although ALA-DAG increases dietary fat utilization as energy in rodents, its effects in humans are not known. The present study was a randomized, placebo-controlled, double-blind, crossover intervention trial performed to clarify the effect of ALA-DAG on postprandial energy metabolism in humans. Nineteen healthy subjects participated in this study, and postprandial energy metabolism was evaluated using indirect calorimetry followed by 14-d repeated pre-consumption of TAG (rapeseed oil) as a control or ALA-DAG. As a primary outcome, ALA-DAG induced significantly higher postprandial fat oxidation than TAG. As a secondary outcome, carbohydrate oxidation tended to be decreased. In addition, postprandial energy expenditure was significantly increased by ALA-DAG compared to TAG. These findings suggest that daily ALA-DAG consumption stimulates dietary fat utilization as energy after a meal, as well as greater diet induced thermogenesis in healthy humans. In conclusion, repeated consumption of ALA-DAG enhanced postprandial fat metabolism after a meal, which may partially explain its visceral fat area-reducing effect. PMID:27430386

  20. Dynamic analysis of marine risers with vortex excitation

    SciTech Connect

    Nordgren, R.P.

    1982-03-01

    The basic equations for nonplanar transverse vibrations of marine risers are derived from the theory of elastic rods. A numerical method is developed for solution of the equations by time integration. Spatial discretization is accomplished by a hybrid finite element method. Vortex excitation is modeled by the coupled wake oscillator proposed by Iwan and Blevins. The vortex oscillator equations are integrated numerically in time along with the riser equations. By way of example, several typical riser problems are analyzed including forced vibration and vortex-induced vibration.

  1. Viscosity and compressibility of diacylglycerol under high pressure

    NASA Astrophysics Data System (ADS)

    Malanowski, Aleksander; Rostocki, A. J.; Kiełczyński, P.; Szalewski, M.; Balcerzak, A.; Kościesza, R.; Tarakowski, R.; Ptasznik, S.; Siegoczyński, R. M.

    2013-03-01

    The influence of high pressure on viscosity and compressibility of diacylglycerol (DAG) oil has been presented in this paper. The investigated DAG oil was composed of 82% of DAGs and 18% TAGs (triacylglycerols). The dynamic viscosity of DAG was investigated as a function of the pressure up to 400 MPa. The viscosity was measured by means of the surface acoustic wave method, where the acoustic waveguides were used as sensing elements. As the pressure was rising, the larger ultrasonic wave attenuation was observed, whereas amplitude decreased with the liquid viscosity augmentation. Measured changes of physical properties were most significant in the pressure range near the phase transition. Deeper understanding of DAG viscosity and compressibility changes versus pressure could shed more light on thermodynamic properties of edible oils.

  2. Diacylglycerol Kinases in the Coordination of Synaptic Plasticity

    PubMed Central

    Lee, Dongwon; Kim, Eunjoon; Tanaka-Yamamoto, Keiko

    2016-01-01

    Synaptic plasticity is activity-dependent modification of the efficacy of synaptic transmission. Although, detailed mechanisms underlying synaptic plasticity are diverse and vary at different types of synapses, diacylglycerol (DAG)-associated signaling has been considered as an important regulator of many forms of synaptic plasticity, including long-term potentiation (LTP) and long-term depression (LTD). Recent evidences indicate that DAG kinases (DGKs), which phosphorylate DAG to phosphatidic acid to terminate DAG signaling, are important regulators of LTP and LTD, as supported by the results from mice lacking specific DGK isoforms. This review will summarize these studies and discuss how specific DGK isoforms distinctly regulate different forms of synaptic plasticity at pre- and postsynaptic sites. In addition, we propose a general role of DGKs as coordinators of synaptic plasticity that make local synaptic environments more permissive for synaptic plasticity by regulating DAG concentration and interacting with other synaptic proteins.

  3. Weight loss effect of dietary diacylglycerol in obese dogs.

    PubMed

    Umeda, T; Bauer, J E; Otsuji, K

    2006-06-01

    Obesity in dogs and cats have been increasingly recognized in recent years. Because obesity underlies various diseases, pet owners and veterinarians have an important responsibility to help animals lose weight and maintain their health. Diet therapy, however, is typically based on limited calorie intake and animals may suffer stress from hunger and this is also a concern to animal owners. For this reason, many clients drop out of weight control programmes. In the present study, we focused on dietary diacylglycerol (DAG) as a potentially effective ingredient for canine weight control without caloric restriction. We replaced a portion of the fat in dog food with either DAG or triacylglycerol (TAG), referred to as DAG or TAG diets here, and fed overweight beagle dogs (body condition score of 4 or higher) with either the DAG or TAG diet for a 6-week period. Results indicated that, even though the food composition other than fat type were identical, dogs fed the DAG diet showed a statistically significant reduction in body weight averaging a 2.3% reduction within 6 weeks while the TAG-fed dogs maintained their obese body weights. In addition, the DAG group also showed a reduction in body fat content, serum triglyceride and total cholesterol concentrations. These results suggest the possibility of developing a pet food using DAG to control weight and serum lipid levels without compromising caloric intake. PMID:16684141

  4. Effect of diacylglycerol supplementation on fasting serum triacylglycerol concentration: a meta-analysis.

    PubMed

    Wang, Wenliang; Xu, Tongcheng; Li, Xia; Zhu, Qingjun; Cheng, Anwei; Du, Fangling; Li, Duo

    2010-12-01

    Diacylglycerol (DAG) supplementation has been shown to be associated with the reduction of fasting serum triacylglycerol (TAG) concentration, although the extent of the association is uncertain. We quantitatively examined the effect of dietary DAG on fasting serum TAG concentration by conducting a meta-analysis of randomized controlled trials. Potential papers were searched from electronic databases of Medline, Embase and Cochrane Library. Information was extracted and the net change of fasting serum TAG concentration was used as the primary outcome to examine the effect of DAG in Review Manager 4.2. Six papers with seven independent studies (298 subjects) were included into the statistic pooling. Meta-analysis with random effect model showed that DAG did not reduce the fasting serum TAG concentration (WMD: -0.07 mmol/L; 95% CI: -0.21 to 0.08 mmol/L; P = 0.37). Sensitivity analysis indicated the robustness of overall results. Fail-safe number analysis indicated that 18 studies with positive effect were necessary to reverse the reported non-significant efficacy of DAG. Weight estimation analysis indicated that the effect of DAG was influenced to some extent by the initial fasting serum TAG concentration. In conclusion, DAG supplementation did not reduce the fasting serum TAG concentration significantly compared with TAG, but some effects were suggested in diabetic patients with hypertriglyceridemia. PMID:21104449

  5. Characterization and quantification of diacylglycerol species in biological extracts after one-step derivatization: a shotgun lipidomics approach.

    PubMed

    Wang, Miao; Hayakawa, Jun; Yang, Kui; Han, Xianlin

    2014-02-18

    Diacylglycerols (DAGs) are important intermediates of lipid metabolism and cellular signaling. It is well-known that the mass levels of DAG are altered under disease states. Therefore, quantitative analysis of DAGs in biological samples can provide critical information to uncover underlying mechanisms of various cellular functional disorders. Although great efforts on the analysis of individual DAG species have recently been made by utilizing mass spectrometry with or without derivatization, cost-effective and high throughput methodologies for identification and quantification of all DAG species including regioisomers, particularly in an approach of shotgun lipidomics, are still missing. Herein, we described a novel method for directly identifying and quantifying DAG species including regioisomers present in lipid extracts of biological samples after facile one-step derivatization with dimethylglycine based on the principles of multidimensional mass spectrometry-based shotgun lipidomics. The established method provided substantial sensitivity (low limit of quantification at amol/μL), high specificity, and broad linear dynamics range (2500-fold) without matrix effects. By exploiting this novel method, we revealed a 16-fold increase of total DAG mass in the livers of ob/ob mice compared to their wild type controls at 4 months of age (an insulin-resistant state) versus a 5-fold difference between 3 month old mice (with normal insulin). These results demonstrated the importance and power of the method for studying biochemical mechanisms underpinning disease states. PMID:24432906

  6. Diacylglycerol mediates regulation of TASK potassium channels by Gq-coupled receptors.

    PubMed

    Wilke, Bettina U; Lindner, Moritz; Greifenberg, Lea; Albus, Alexandra; Kronimus, Yannick; Bünemann, Moritz; Leitner, Michael G; Oliver, Dominik

    2014-01-01

    The two-pore domain potassium (K2P) channels TASK-1 (KCNK3) and TASK-3 (KCNK9) are important determinants of background K(+) conductance and membrane potential. TASK-1/3 activity is regulated by hormones and transmitters that act through G protein-coupled receptors (GPCR) signalling via G proteins of the Gαq/11 subclass. How the receptors inhibit channel activity has remained unclear. Here, we show that TASK-1 and -3 channels are gated by diacylglycerol (DAG). Receptor-initiated inhibition of TASK required the activity of phospholipase C, but neither depletion of the PLC substrate PI(4,5)P2 nor release of the downstream messengers IP3 and Ca(2+). Attenuation of cellular DAG transients by DAG kinase or lipase suppressed receptor-dependent inhibition, showing that the increase in cellular DAG-but not in downstream lipid metabolites-mediates channel inhibition. The findings identify DAG as the signal regulating TASK channels downstream of GPCRs and define a novel role for DAG that directly links cellular DAG dynamics to excitability. PMID:25420509

  7. P2Y₁ receptor-dependent diacylglycerol signaling microdomains in β cells promote insulin secretion.

    PubMed

    Wuttke, Anne; Idevall-Hagren, Olof; Tengholm, Anders

    2013-04-01

    Diacylglycerol (DAG) controls numerous cell functions by regulating the localization of C1-domain-containing proteins, including protein kinase C (PKC), but little is known about the spatiotemporal dynamics of the lipid. Here, we explored plasma membrane DAG dynamics in pancreatic β cells and determined whether DAG signaling is involved in secretagogue-induced pulsatile release of insulin. Single MIN6 cells, primary mouse β cells, and human β cells within intact islets were transfected with translocation biosensors for DAG, PKC activity, or insulin secretion and imaged with total internal reflection fluorescence microscopy. Muscarinic receptor stimulation triggered stable, homogenous DAG elevations, whereas glucose induced short-lived (7.1 ± 0.4 s) but high-amplitude elevations (up to 109 ± 10% fluorescence increase) in spatially confined membrane regions. The spiking was mimicked by membrane depolarization and suppressed after inhibition of exocytosis or of purinergic P2Y₁, but not P2X receptors, reflecting involvement of autocrine purinoceptor activation after exocytotic release of ATP. Each DAG spike caused local PKC activation with resulting dissociation of its substrate protein MARCKS from the plasma membrane. Inhibition of spiking reduced glucose-induced pulsatile insulin secretion. Thus, stimulus-specific DAG signaling patterns appear in the plasma membrane, including distinct microdomains, which have implications for the kinetic control of exocytosis and other membrane-associated processes. PMID:23299857

  8. Directed Acyclic Graphs for Oral Disease Research.

    PubMed

    Akinkugbe, A A; Sharma, S; Ohrbach, R; Slade, G D; Poole, C

    2016-07-01

    Directed acyclic graphs (DAGs) are nonparametric graphical tools used to depict causal relations in the epidemiologic assessment of exposure-outcome associations. Although their use in dental research was first advocated in 2002, DAGs have yet to be widely adopted in this field. DAGs help identify threats to causal inference such as confounders, bias due to subject selection, and inappropriate handling of missing data. DAGs can also inform the data analysis strategy based on relations among variables depicted on it. This article uses the example of a study of temporomandibular disorders (TMDs), investigating causal effects of facial injury on subsequent risk of TMD. We illustrate how DAGs can be used to identify 1) potential confounders, 2) mediators and the consequences of attempt to estimate direct causal effects, 3) colliders and the consequences of conditioning on colliders, and 4) variables that are simultaneously mediators and confounders and the consequences of adjustment for such variables. For example, one DAG shows that statistical adjustment for the pressure pain threshold would necessarily bias the causal relation between facial injury and TMD. Finally, we discuss the usefulness of DAGs during study design, subject selection, and choosing variables to be measured in a study. PMID:27000052

  9. Progesterone and the zona pellucida activate different transducing pathways in the sequence of events leading to diacylglycerol generation during mouse sperm acrosomal exocytosis.

    PubMed Central

    Murase, T; Roldan, E R

    1996-01-01

    We tested the involvement of protein tyrosine kinase and G-protein transducing pathways in the formation of diacylglycerol (DAG) during exocytosis in mouse spermatozoa. In capacitated spermatozoa, stimulation with solubilized zona pellucida (ZP) or progesterone led to the formation of DAG and to exocytosis of the acrosomal granule. Stimulation of DAG formation and exocytosis by ZP were inhibited in a concentration-dependent fashion by pre-exposure to tyrphostin A48, a protein tyrosine kinase inhibitor. These ZP-induced responses were also reduced in a concentration-dependent manner by prior incubation with pertussis toxin, a G-protein (Gi class) inhibitor. On the other hand, generation of DAG and exocytosis triggered by progesterone were inhibited if spermatozoa were preincubated with different concentrations of tyrphostin A48, but were not affected by pre-exposure to pertussis toxin. Progesterone acts on at least two novel surface receptors, one being a gamma-aminobutyric acid (GABA) type A (GABAA)-like receptor. Transducing mechanisms coupled to this receptor were tested directly by stimulating spermatozoa with GABA. Treatment of capacitated spermatozoa with GABA resulted in DAG formation and exocytosis. These responses were not seen when cells were preincubated with tyrphostin A48. Pertussis toxin, however, did not affect the generation of DAG and exocytosis triggered by GABA, in agreement with results obtained using progesterone. Taken together, these results indicate that DAG formation during acrosomal exocytosis is differentially regulated by transducing pathways activated by oocyte-associated agonists. PMID:9003394

  10. Characterization and Quantification of Diacylglycerol Species in Biological Extracts after One-step Derivatization: A Shotgun Lipidomics Approach

    PubMed Central

    Wang, Miao; Hayakawa, Jun; Yang, Kui; Han, Xianlin

    2014-01-01

    Diacylglycerols (DAGs) are important intermediates of lipid metabolism and cellular signaling. It is well known that the mass levels of DAG are altered under disease states. Therefore, quantitative analysis of DAGs in biological samples can provide critical information to uncover underlying mechanisms of various cellular functional disorders. Although great efforts on the analysis of individual DAG species have recently been made by utilizing mass spectrometry with or without derivatization, cost effective and high throughput methodology for identification and quantification of all DAG species including regioisomers, particularly in an approach of shotgun lipidomics, are still missing. Herein, we described a novel method for directly identifying and quantifying DAG species including regioisomers present in lipid extracts of biological samples after facile one-step derivatization with dimethylglycine based on the principles of multi-dimensional mass spectrometry-based shotgun lipidomics. The established method provided substantial sensitivity (low limit of quantification at amol/µl), high specificity, and broad linear dynamics range (2,500 folds) without matrix effects. By exploiting this novel method, we revealed a 16-fold increase of total DAG mass in the livers of ob/ob mice compared to their wild type controls at 4 months of age (an insulin-resistant state) vs. a 5-fold difference between 3-month old mice (with normal insulin). These results demonstrated the importance and power of the method for studying biochemical mechanisms underpinning disease states. PMID:24432906

  11. Relationships among certain joint constitutive models.

    SciTech Connect

    Segalman, Daniel Joseph; Starr, Michael James

    2004-09-01

    In a recent paper, Starr and Segalman demonstrated that any Masing model can be represented as a parallel-series Iwan model. A preponderance of the constitutive models that have been suggested for simulating mechanical joints are Masing models, and the purpose of this discussion is to demonstrate how the Iwan representation of those models can yield insight into their character. In particular, this approach can facilitate a critical comparison among numerous plausible constitutive models. It is explicitly shown that three-parameter models such as Smallwood's (Ramberg-Osgood) calculate parameters in such a manner that macro-slip is not an independent parameter, yet the model admits macro-slip. The introduction of a fourth parameter is therefore required. It is shown that when a macro-slip force is specified for the Smallwood model the result is a special case of the Segalman four-parameter model. Both of these models admit a slope discontinuity at the inception of macro-slip. A five-parameter model that has the beneficial features of Segalman's four-parameter model is proposed. This model manifests a force-displacement curve having a continuous first derivative.

  12. Diacylglycerol kinase ζ (DGKζ) is a critical regulator of bone homeostasis via modulation of c-Fos levels in osteoclasts†

    PubMed Central

    Zamani, Ali; Decker, Corinne; Cremasco, Viviana; Hughes, Lindsey; Novack, Deborah V.; Faccio, Roberta

    2015-01-01

    Increased diacylglycerol (DAG) levels are observed in numerous pathologies, including conditions associated with bone loss. However, the effects of DAG accumulation on the skeleton have never been directly examined. Because DAG is strictly controlled by tissue specific diacylglycerol kinases (DGKs), we sought to examine the biological consequences of DAG accumulation on bone homeostasis by genetic deletion of DGKζ, a highly expressed DGK isoform in osteoclasts (OCs). Strikingly, DGKζ−/− mice are osteoporotic due to a marked increase in OC numbers. In vitro, DGKζ−/− bone marrow macrophages (BMMs) form more numerous, larger and highly resorptive OCs. Surprisingly, while increased DAG levels do not alter RANK/RANKL osteoclastogenic pathway, DGKζ deficiency increases responsiveness to the proliferative and pro-survival cytokine M-CSF. We find that M-CSF is responsible for increased DGKζ−/− OC differentiation by promoting higher expression of the transcription factor c-Fos, and c-Fos knockdown in DGKζ−/− cultures dose-dependently reduces OC differentiation. Using a c-Fos luciferase reporter assay lacking the TRE responsive element, we also demonstrate that M-CSF induces optimal c-Fos expression through DAG production. Finally, to demonstrate the importance of the M-CSF/DGKζ/DAG axis on regulation of c-Fos during osteoclastogenesis, we turned to PLCγ2+/− BMMs, which have reduced DAG levels and form fewer OCs due to impaired expression of the master regulator of osteoclastogenesis NFATc1 and c-Fos. Strikingly, genetic deletion of DGKζ in PLCγ2+/− mice rescues OC formation and normalizes c-Fos levels without altering NFATc1 expression. To our knowledge, this is the first report implicating M-CSF/DGKζ/DAG axis as a critical regulator of bone homeostasis via its actions on OC differentiation and c-Fos expression. PMID:25891971

  13. Autocrine Signaling Underlies Fast Repetitive Plasma Membrane Translocation of Conventional and Novel Protein Kinase C Isoforms in β Cells*

    PubMed Central

    Wuttke, Anne; Yu, Qian; Tengholm, Anders

    2016-01-01

    PKC signaling has been implicated in the regulation of many cell functions, including metabolism, cell death, proliferation, and secretion. Activation of conventional and novel PKC isoforms is associated with their Ca2+- and/or diacylglycerol (DAG)-dependent translocation to the plasma membrane. In β cells, exocytosis of insulin granules evokes brief (<10 s) local DAG elevations (“spiking”) at the plasma membrane because of autocrine activation of P2Y1 purinoceptors by ATP co-released with insulin. Using total internal reflection microscopy, fluorescent protein-tagged PKCs, and signaling biosensors, we investigated whether DAG spiking causes membrane recruitment of PKCs and whether different classes of PKCs show characteristic responses. Glucose stimulation of MIN6 cells triggered DAG spiking with concomitant repetitive translocation of the novel isoforms PKCδ, PKCϵ, and PKCη. The conventional PKCα, PKCβI, and PKCβII isoforms showed a more complex pattern with both rapid and slow translocation. K+ depolarization-induced PKCϵ translocation entirely mirrored DAG spiking, whereas PKCβI translocation showed a sustained component, reflecting the subplasma membrane Ca2+ concentration ([Ca2+]pm), with additional effect during DAG spikes. Interference with DAG spiking by purinoceptor inhibition prevented intermittent translocation of PKCs and reduced insulin secretion but did not affect [Ca2+]pm elevation or sustained PKCβI translocation. The muscarinic agonist carbachol induced pronounced transient PKCβI translocation and sustained recruitment of PKCϵ. When rise of [Ca2+]pm was prevented, the carbachol-induced DAG and PKCϵ responses were somewhat reduced, but PKCβI translocation was completely abolished. We conclude that exocytosis-induced DAG spikes efficiently recruit both conventional and novel PKCs to the β cell plasma membrane. PKC signaling is thus implicated in autocrine regulation of β cell function. PMID:27226533

  14. Autocrine Signaling Underlies Fast Repetitive Plasma Membrane Translocation of Conventional and Novel Protein Kinase C Isoforms in β Cells.

    PubMed

    Wuttke, Anne; Yu, Qian; Tengholm, Anders

    2016-07-15

    PKC signaling has been implicated in the regulation of many cell functions, including metabolism, cell death, proliferation, and secretion. Activation of conventional and novel PKC isoforms is associated with their Ca(2+)- and/or diacylglycerol (DAG)-dependent translocation to the plasma membrane. In β cells, exocytosis of insulin granules evokes brief (<10 s) local DAG elevations ("spiking") at the plasma membrane because of autocrine activation of P2Y1 purinoceptors by ATP co-released with insulin. Using total internal reflection microscopy, fluorescent protein-tagged PKCs, and signaling biosensors, we investigated whether DAG spiking causes membrane recruitment of PKCs and whether different classes of PKCs show characteristic responses. Glucose stimulation of MIN6 cells triggered DAG spiking with concomitant repetitive translocation of the novel isoforms PKCδ, PKCϵ, and PKCη. The conventional PKCα, PKCβI, and PKCβII isoforms showed a more complex pattern with both rapid and slow translocation. K(+) depolarization-induced PKCϵ translocation entirely mirrored DAG spiking, whereas PKCβI translocation showed a sustained component, reflecting the subplasma membrane Ca(2+) concentration ([Ca(2+)]pm), with additional effect during DAG spikes. Interference with DAG spiking by purinoceptor inhibition prevented intermittent translocation of PKCs and reduced insulin secretion but did not affect [Ca(2+)]pm elevation or sustained PKCβI translocation. The muscarinic agonist carbachol induced pronounced transient PKCβI translocation and sustained recruitment of PKCϵ. When rise of [Ca(2+)]pm was prevented, the carbachol-induced DAG and PKCϵ responses were somewhat reduced, but PKCβI translocation was completely abolished. We conclude that exocytosis-induced DAG spikes efficiently recruit both conventional and novel PKCs to the β cell plasma membrane. PKC signaling is thus implicated in autocrine regulation of β cell function. PMID:27226533

  15. Multiple sources of 1,2-diacylglycerol in isolated rat pancreatic acini stimulated by cholecystokinin. Involvement of phosphatidylinositol bisphosphate and phosphatidylcholine hydrolysis

    SciTech Connect

    Matozaki, T.; Williams, J.A. )

    1989-09-05

    Changes in the cellular content of 1,2-diacylglycerol (DAG) in isolated rat pancreatic acini in response to agonist stimulation were studied using a sensitive mass assay. When acini were stimulated by 10 nM COOH-terminal cholecystokinin-octapeptide (CCK8), the increase in DAG was biphasic, consisting of an early peak at 5 s and a second, larger, gradual increase that was maximal by 15 min. The basal level of DAG in acini was 1.04 nmol/mg of protein, which was increased to 1.24 nmol/mg of protein at 5 s and 2.76 nmol/mg of protein at 30 min. In comparison, the increase in DAG stimulated by 30 pM CCK8, a submaximal concentration for amylase release, was monophasic, increasing without an early peak but sustained to 60 min. Other Ca2+-mobilizing secretagogues such as carbamylcholine and bombesin increased DAG in acini, whereas vasoactive intestinal peptide, which acts to increase cAMP, had no effect. Phorbol ester and Ca2+ ionophore also stimulated DAG production. Analysis of the mass level of inositol 1,4,5-trisphosphate (1,4,5-IP3) showed that the generation of 1,4,5-IP3 stimulated by 10 nM CCK8 peaked at 5 s, a finding consistent with the early peak of DAG. The basal level was 4.7 pmol/mg of protein, which was increased to 144.6 pmol/mg of protein at 5 s by 10 nM CCK8. The levels of 1,4,5-IP3 then returned toward basal in contrast to the gradual and sustained increase of DAG. The dose dependencies of 1,4,5-IP3 and DAG formation at 5 s with respect to CCK8 were almost identical. This suggests that phosphatidylinositol 4,5-bisphosphate hydrolysis is a major source of the early increase in DAG but not of the sustained increase in DAG. Therefore, a possible contribution of phosphatidylcholine hydrolysis to DAG formation was examined utilizing acini prelabeled with (3H)choline. CCK8 (1 nM) maximally increased (3H)choline metabolite release by 133% of control at 30 min.

  16. Application of solvent engineering to optimize lipase-catalyzed 1,3-diglyacylcerols by mixture response surface methodology.

    PubMed

    Liao, Hui-Fen; Tsai, Wei-Chuan; Chang, Shu-Wei; Shieh, Chwen-Jen

    2003-11-01

    1,3-Diacylglycerol (DAG) has been introduced in Japan as a cooking oil under the trade name of Econa to reduce body fat accumulation. Solvent engineering was applied to determine the optimum solvent mixtures for the lipase-catalyzed synthesis of 1,3-DAG by mixture response surface methodology. n-Hexane was required to maintain the lipase activity and the product selectivity could be adjusted by changing the hydrophobicity of reaction medium. The optimum yield (approximately 40%) of 1,3-DAG synthesis was obtained with n-hexane/octane (1:1, v/v). PMID:14677712

  17. Seven Modeling Perspectives on Teaching and Learning: Some Interrelations and Cognitive Effects

    ERIC Educational Resources Information Center

    Easley, J. A., Jr.

    1977-01-01

    The categories of models associated with the seven perspectives are designated as combinatorial models, sampling models, cybernetic models, game models, critical thinking models, ordinary language analysis models, and dynamic structural models. (DAG)

  18. Quantification of diacylglycerol by mass spectrometry.

    PubMed

    vom Dorp, Katharina; Dombrink, Isabel; Dörmann, Peter

    2013-01-01

    Diacylglycerol (DAG) is an important intermediate of lipid metabolism and a component of phospholipase C signal transduction. Quantification of DAG in plant membranes represents a challenging task because of its low abundance. DAG can be measured by direct infusion mass spectrometry (MS) on a quadrupole time-of-flight mass spectrometer after purification from the crude plant lipid extract via solid-phase extraction on silica columns. Different internal standards are employed to compensate for the dependence of the MS and MS/MS signals on the chain length and the presence of double bonds in the acyl moieties. Thus, using a combination of single MS and MS/MS experiments, quantitative results for the different molecular species of DAGs from Arabidopsis can be obtained. PMID:23681522

  19. Prolactin-stimulated ornithine decarboxylase induction in rat hepatocytes: Coupling to diacylglycerol generation and protein kinase C

    SciTech Connect

    Buckley, A.R.; Buckley, D.J. )

    1991-01-01

    The trophic effects of prolactin (PRL) in rat liver have been linked to activation of protein kinase C (PKC). Since alterations in PKC activity imply its activation by 1,2-diacylglycerol (DAG), we tested whether PRL treatment stimulated DAG generation coupled to induction of a growth response in primary hepatocytes. Addition of PRL to hepatocyte cultures significantly increased ({sup 3}H)-glycerol incorporation into DAG within 5 minutes which was followed by a loss of cytosolic PKC activity by 10 minutes. Prolactin also significantly enhanced radiolabel incorporation into triacylglycerol and phospholipids within 10 minutes and induced ODC activity at 6 hours. Therefore, prolactin-stimulated alterations in PKC activity are preceded by enhanced DAG generation. Moreover, these events appear to be coupled to PRL-stimulated entry of hepatocytes into cell cycle.

  20. Role of diacylglycerol kinase in cellular regulatory processes: a new regulator for cardiomyocyte hypertrophy.

    PubMed

    Takeishi, Yasuchika; Goto, Kaoru; Kubota, Isao

    2007-09-01

    Diacylglycerol (DAG) kinase (DGK) phosphorylates and converts DAG to phosphatidic acid. DGK regulates cellular DAG levels and attenuates DAG signaling. The 10 mammalian DGK isoforms have been identified to date. In cardiac myocytes, DGKalpha, epsilon, and zeta are expressed, and DGKzeta is the predominant isoform. DGKzeta inhibits protein kinase C (PKC) activation and subsequent hypertrophic programs in response to endothelin-1 (ET-1) in neonatal rat cardiomyocytes. DGKzeta blocks cardiac hypertrophy induced by G protein-coupled receptor agonists and pressure overload in vivo. DGKzeta attenuates ventricular remodeling and improves survival after myocardial infarction. These data provide a novel insight for subcellular mechanisms of cardiac hypertrophy and heart failure, and DGKzeta may be a new therapeutic target to prevent cardiac hypertrophy and progression to heart failure. PMID:17659347

  1. Planning for Future Outcomes: One District and Beyond

    ERIC Educational Resources Information Center

    Tuckman, B. W.; And Others

    1977-01-01

    The story of one school district that developed a strategy of comprehensive planning to maximize resources and to increase the involvement of teachers, administrators and parents in the process of goal-setting and assessment. (Author/DAG)

  2. Directory of the Florida Motion Picture and Television Industry.

    ERIC Educational Resources Information Center

    Florida State Dept. of Commerce, Tallahassee. Div. of Economic Development.

    Designed to assist the motion picture or television producer, this directory lists organizations (producers, distributors, talent agencies, laboratories, etc.) by geographical section in Florida. Each entry includes the company address, telephone, services available, a contact person, and credits. (DAG)

  3. A novel mechanism for acetylcholine to generate diacylglycerol in brain

    SciTech Connect

    Qian, Z.; Drewes, L.R. )

    1990-03-05

    The classical scheme involving inositol phospholipid breakdown by phospholipase C as the sole source of diacylglycerol (DAG) has recently been challenged by evidence that phosphatidylcholine (PC) is an alternative source. In synaptic membranes of canine cerebral cortex, cholinergic agonists caused rapid accumulation of ({sup 3}H)phosphatidic acid (PA) from ({sup 3}H)PC within 15 s, whereas (3H)DAG formation showed a transient lag period before becoming elevated and then exceeding the amount of ({sup 3}H)PA. Additional evidence shows that DAG is produced from PC by the action of phospholipase D to yield PA, which is further dephosphorylated to DAG by PA phosphatase. Our results indicate that this muscarinic acetylcholine receptor-regulated PC phospholipase D-PA phosphatase pathway may be a novel mechanism in cell signal transduction processes for activation of protein kinase C in brain.

  4. The MICA Project: A Sophisticated Operational CMI System

    ERIC Educational Resources Information Center

    Chapin, John D.; Patterson, Jerry L.

    1977-01-01

    Managed Instruction with Computer Assistance (MICA) used a teacher-designed math curriculum as a pilot project for a computer managed instruction (CMI) system designed to provide a means of implementing individualization. (DAG)

  5. Mechanism of protein kinase C activation by phosphatidylinositol 4,5-bisphosphate

    SciTech Connect

    Lee, Myungho; Bell, R.M. )

    1991-01-29

    The mechanism of protein kinase C (PKC) activation by phosphatidylinositol 4,5-bisphosphate (PIP{sub 2}), phosphatidylinositol 4-monophosphate (PIP), and phosphatidylinositol (PI) was investigated by using Triton X-100 mixed micellar methods. The activation of PKC by PIP{sub 2}, for which maximal activity was 60% of that elicited by sn-1,2-diacylglycerol (DAG), was similar to activation by DAG in several respects: (1) activation by PIP{sub 2} and DAG required phosphatidylserine (PS) as a phospholipid cofactor, (2) PIP{sub 2} and DAG reduced the concentration of Ca{sup 2+} and PS required for activation, (3) the concentration dependences of activation by PIP{sub 2} and DAG depended on the concentration of PS, and (4) PIP{sub 2} and DAG complemented one another to achieve maximal activation. On the other hand, PIP{sub 2} activation of the PKC differed from activation by DAG in several respects. With increasing concentrations of PIP{sub 2}, (1) the optimal concentration of PS required was constant at 12 mol%, (2) the maximal activity at 12 mol% PS increased, and (3) the cooperativity for PS decreased. PIP{sub 2} did not inhibit ({sup 3}H)phorbol 12,13-dibutyrate (PDBu) binding of PKC at saturating levels of PS; however, at subsaturating levels of PS, PIP{sub 2} enhanced ({sup 3}H)PDBu binding by acting as a phospholipid cofactor. PIP did not function as an activator but served as a phospholipid cofactor in the presence of PS. These data establish that PIP{sub 2}, PIP, and PI can function to spare, in part, the PS phospholipid cofactor requirement of PKC, and they demonstrate that PIP{sub 2} but not PIP and PI can function as a lipid activator of PKC by mechanisms distinct from those of DAG and phorbol esters.

  6. Berberine treatment attenuates the palmitate-mediated inhibition of glucose uptake and consumption through increased 1,2,3-triacyl-sn-glycerol synthesis and accumulation in H9c2 cardiomyocytes.

    PubMed

    Chang, Wenguang; Chen, Li; Hatch, Grant M

    2016-04-01

    Dysfunction of lipid metabolism and accumulation of 1,2-diacyl-sn-glycerol (DAG) may be a key factor in the development of insulin resistance in type 2 diabetes. Berberine (BBR) is an isoquinoline alkaloid extract that has shown promise as a hypoglycemic agent in the management of diabetes in animal and human studies. However, its mechanism of action is not well understood. To determine the effect of BBR on lipid synthesis and its relationship to insulin resistance in H9c2 cardiomyocytes, we measured neutral lipid and phospholipid synthesis and their relationship to glucose uptake. Compared with controls, BBR treatment stimulated 2-[1,2-(3)H(N)]deoxy-D-glucose uptake and consumption in palmitate-mediated insulin resistant H9c2 cells. The mechanism was though an increase in protein kinase B (AKT) activity and GLUT-4 glucose transporter expression. DAG accumulated in palmitate-mediated insulin resistant H9c2 cells and treatment with BBR reduced this DAG accumulation and increased accumulation of 1,2,3-triacyl-sn-glycerol (TAG) compared to controls. Treatment of palmitate-mediated insulin resistant H9c2 cells with BBR increased [1,3-(3)H]glycerol and [1-(14)C]glucose incorporation into TAG and reduced their incorporation into DAG compared to control. In addition, BBR treatment of these cells increased [1-(14)C]palmitic acid incorporation into TAG and decreased its incorporation into DAG compared to controls. BBR treatment did not alter phosphatidylcholine or phosphatidylethanolamine synthesis. The mechanism for the BBR-mediated decreased precursor incorporation into DAG and increased incorporation into TAG in palmitate-incubated cells was an increase in DAG acyltransferase-2 activity and its expression and a decrease in TAG hydrolysis. Thus, BBR treatment attenuates palmitate-induced reduction in glucose uptake and consumption, in part, through reduction in cellular DAG levels and accumulation of TAG in H9c2 cells. PMID:26774040

  7. The Effects of a High Fat Diet Containing Diacylglycerol on Bone in C57BL/6J Mice

    PubMed Central

    Choi, Han Seok; Park, Su Jin; Lee, Zang Hee

    2015-01-01

    Purpose In epidemiologic and animal studies, a high fat diet (HFD) has been shown to be associated with lower bone mineral density (BMD) and a higher risk of osteoporotic fractures. Meanwhile, consuming a HFD containing diacylglycerol (DAG) instead of triacylglycerol (TAG) is known to offer metabolically beneficial effects of reductions in body weight and abdominal fat. The purpose of this study was to investigate the effects of a HFD containing DAG (HFD-DAG) on bone in mice. Materials and Methods Four-week-old male C57BL/6J mice (n=39) were divided into three weight-matched groups based on diet type: a chow diet group, a HFD containing TAG (HFD-TAG) group, and a HFD-DAG group. After 20 weeks, body composition and bone microstructure were analyzed using dual energy X-ray absorptiometry and micro-computed tomography. Reverse transcription-polymerase chain reaction (PCR) and real-time PCR of bone marrow cells were performed to investigate the expressions of transcription factors for osteogenesis or adipogenesis. Results The HFD-DAG group exhibited lower body weight, higher BMD, and superior microstructural bone parameters, compared to the HFD-TAG group. The HFD-DAG group showed increased expression of Runx2 and decreased expression of PPARγ in bone marrow cells, compared to the HFD-TAG group. The HFD-DAG group also had lower levels of plasma glucose, insulin, total cholesterol, and triglyceride than the HFD-TAG group. Conclusion Compared to HFD-TAG, HFD-DAG showed beneficial effects on bone and bone metabolism in C57BL/6J mice. PMID:26069116

  8. Production of Diacylglycerol-enriched Oil by Glycerolysis of Soybean Oil using a Bubble Column Reactor in a Solvent-free System.

    PubMed

    Zhang, Ning; Yang, Xue; Fu, Junning; Chen, Qiong; Song, Ziliang; Wang, Yong

    2016-03-01

    In this study, diacylglycerol-enriched soybean oil (DESO) was synthesized through Lipozyme 435-catalyzed glycerolysis of soybean oil (SO) in a solvent-free system using a modified bubble column reactor. The effects of enzyme load, mole ratio of glycerol to soybean oil, reaction temperature, gas flow and reaction time on DAG production were investigated. The selected conditions were established as being enzyme load of 4 wt% (mass of substrates), glycerol/soybean oil mole ratio of 20:1, reaction temperature of 80°C, gas flow of 10.6 cm/min, and a reaction time of 2.5 h, obtaining the DAG content of 49.4±0.5 wt%. The reusability of Lipozyme 435 was evaluated by monitoring the contents of DAG, monoacylglycerol (MAG) and triacylglycerol (TAG) in 10 consecutive runs. After purified by one-step molecular distillation, the DAG content of 63.5±0.3 wt% was achieved in DESO. The mole ratio of 1, 3-DAG to 1, 2-DAG was 2:1 and the fatty acid composition had no significant difference from that of soybean oil. However, the thermal properties of DESO and SO had considerable differences. Polymorphic form of DESO were mainly the β form and minor amounts of the β' form. Granular aggregation and round-shaped crystals were detected in DESO. PMID:26876674

  9. Self-Assembly and Lipid Interactions of Diacylglycerol Lactone Derivatives Studied at the Air/Water Interface

    PubMed Central

    Philosof-Mazor, Liron; Volinsky, Roman; Comin, Maria J.; Lewin, Nancy E.; Kedei, Noemi; Blumberg, Peter M.; Marquez, Victor E.; Jelinek, Raz

    2009-01-01

    Synthetic diacylglycerol lactones (DAG-lactones) have been shown to be effective modulators of critical cellular signaling pathways. The biological activity of these amphiphilic molecules depends in part upon their lipid interactions within the cellular plasma membrane. This study explores the thermodynamic and structural features of DAG-lactone derivatives and their lipid interactions at the air/water interface. Surface-pressure/area isotherms and Brewster angle microscopy revealed the significance of specific side-groups attached to the terminus of a very rigid 4-(2-phenylethynyl) benzoyl chain of the DAG-lactones, which affected both the self-assembly of the molecules and their interactions with phospholipids. The experimental data highlight the formation of different phases within mixed DAG-lactone/phospholipid monolayers and underscore the relationship between the two components in binary mixtures of different mole ratios. Importantly, the results suggest that DAG-lactones are predominantly incorporated within fluid phospholipid phases rather than in the condensed phases that form, for example, by cholesterol. Moreover, the size and charge of the phospholipid headgroups do not seem to affect DAG-lactone interactions with lipids. PMID:18788772

  10. Determination of physicochemical properties of diacylglycerol oil at high pressure by means of ultrasonic methods.

    PubMed

    Kiełczyński, Piotr; Szalewski, Marek; Balcerzak, Andrzej; Wieja, Krzysztof; Malanowski, Aleksander; Kościesza, Rafał; Tarakowski, Rafał; Rostocki, Aleksander J; Siegoczyński, Ryszard M

    2014-12-01

    The purpose of the paper is to address, using ultrasonic methods, the impact of temperature and pressure on the physicochemical properties of liquids on the example of diacylglycerol (DAG) oil. The paper presents measurements of sound velocity, density and volume of DAG oil sample in the pressure range from atmospheric pressure up to 0.6GPa and at temperatures ranging from 20 to 50°C. Sound speed measurements were performed in an ultrasonic setup with a DAG oil sample located in the high-pressure chamber. An ultrasonic method that uses cross-correlation method to determine the time-of-flight of the ultrasonic pulses through the liquid was employed to measure the sound velocity in DAG oil. This method is fast and reliable tool for measuring sound velocity. The DAG oil density at high pressure was determined from the monitoring of sample volume change. The adiabatic compressibility and isothermal compressibility have been calculated on the basis of experimental data. Discontinuities in isotherms of the sound speed versus pressure point to the existence of phase transitions in DAG oil. The ultrasonic method presented in this study can be applied to investigate the physicochemical parameters of other liquids not only edible oils. PMID:25017363

  11. Positional-Species Composition of Diacylglycerol Acetates from Mature Euonymus Seeds.

    PubMed

    Sidorov, Roman A; Pchelkin, Vasily P; Zhukov, Anatoly V; Tsydendambaev, Vladimir D

    2016-06-01

    The positional-species composition (PSC) of 3-acetyl-1,2-diacyl-sn-glycerols (AcDAGs) from the seeds of mature fruits of 14 species of the genus Euonymus L. was established. The residues of six major fatty acids (FAs), palmitic (P), stearic (St), hexadecenoic (H), octadecenoic (O), linoleic (L), and linolenic (Ln), were present in the AcDAGs. Here, we demonstrated that the profile of PSC of AcDAGs could serve as chemotaxonomic factor to divide euonymus species studied here into groups which completely correlate with the present day systematic of the genus. In particular, the Euonymus section greatly exceeded other sections of the Euonymus subgenus as well as the Kalonymus one in the total levels of AcDAGs positional species having one and two O residues and was characterized by significantly lesser concentrations of species with one and two L residues. Moreover, in seed, AcDAGs of almost all Euonymus species EFL values were slightly higher than EFO ones, but all EFL and EFO values were higher than 1.0, and therefore, it can be concluded that both FAs mainly esterified sn-2-position of the glycerol moiety and saturated FAs residues were always virtually absent in the sn-2 position of Euonymus seed AcDAGs, as it is also the case in nearly all TAGs molecules of plant origin. PMID:27151557

  12. Cosmogenic Records in 18 Ordinary Chondrites from the Dar Al Gani Region, Libya. 2; Radionclides

    NASA Technical Reports Server (NTRS)

    Welten, K. C.; Nishiizumi, K.; Finkel, R. C.; Hillegonds, D. J.; Jull, A. J. T.; Schultz, L.

    2003-01-01

    In the past decade more than 1000 meteorites have been recovered from the Dar al Gani (DaG) plateau in the Libyan part of the Sahara. The geological setting, meteorite pairings and density are described. So far, only a few terrestrial ages are known for DaG meteorites, e.g. 60+/- 20 kyr for the DaG 476 shergottite shower and 80+/- 20 kyr for the lunar meteorite DaG 262. However, from other desert areas, such as Oman, it is known that achondrites may survive much longer than chondritic meteorites, so the ages of these two achondrites may not be representative of the majority of the DaG meteorite collection, of which more than 90% are ordinary chondrites. In this work we report concentrations of the cosmogenic radionuclides, 14C (half-life = 5,730 yr), 41Ca (1.04x10 superscript 5 yr), Cl-36 (3.01x10 superscript 5 yr), Al-26 (7.05x10 superscript 5 yr) and 10Be (1.5x10 superscript 6 yr) to determine the terrestrial ages of DaG meteorites and constrain their pre-atmospheric size and exposure history.

  13. [DIACYLGLYCEROL ACCUMULATION IMPAIRS SHORT-TERM ACTIVATION OF PHOSPHOLIPASE D BY THYROXINE IN THE LIVER CELLS].

    PubMed

    Hassouneh, Loay Kh M

    2015-01-01

    Thyroid hormones (TG) are known modulators of signal transduction. Phospholipase D (PLD) is one of the targets of TG in the stimulated cells. Response of cells to the short-term TG action significantly reduces at old age. Taking into account that diacylglycerol (DAG) accumulation induces the resistance of cells to some of regulatory factors in the target cells the aim of the present study was to determine if DAG content increase in hepatocytes impairs the L-thyroxine (L-T4) short-term action. The experiments were performed in either the [14C]palmitic acid- labeled hepatocytes or [14C]oleic acid-pre-labeled liver cells of 3- and 24-month-old rats. To study the short-term L-T4 action on cells the PLD activation was determined. The DAG production and content in hepatocytes significantly increased at old age and in the young cells pre-treated with palmitic acid. The reduction of DAG level in cells by means of DAG-kinase activator, alfa-tocoferol acetate, or long-term L-T4 treatment improved the short-term hormone action. The above data have indicated that DAG play important role in the L-T4 PLD regulation. The cross-talk between classic and non-genomic pathways of TG regulation of lipid metabolism has been determined. PMID:26387163

  14. Impact of diacylglycerol and monoacylglycerol on the physical and chemical properties of stripped soybean oil.

    PubMed

    Chen, Bingcan; McClements, David Julian; Decker, Eric Andrew

    2014-01-01

    In this study, we determined the effect of diacylglycerol (DAG) and monoacylglycerol (MAG) on the oxidative stability of stripped soybean oil (SSO) and on the antioxidative effectiveness of α-tocopherol in SSO. We also examined the influence of DAG and MAG on the physical properties of SSO. DAG (0-2.5wt%) had little effect on the chemical stability of SSO and on the antioxidative activity of 40μM α-tocopherol in SSO (55°C). MAG (0-2.5wt%) had no remarkable impact on the chemical stability of SSO. The addition of MAG (0.5wt%) suppressed the effectiveness of α-tocopherol (40μM) in SSO. The addition of DAG did not cause an appreciable change in the interfacial tension (IFT) of SSO, indicating that it was not strongly surface active. MAG significantly decreased the interfacial tension of SSO, due to its strong surface active properties. Wide angle X-ray scattering (WAXS) analysis showed that DAG did not alter the structured organisation of SSO, which remained in an amorphous form, whereas MAG led to strong scattering, indicating the formation of crystals. The physical properties of DAG and MAG in the SSO may be related to the chemical stability of SSO and the effectiveness of antioxidants incorporated. These results can be used to better understand the physicochemical mechanisms by which minor components impact oxidation of bulk oils. PMID:24001854

  15. Glycidol exposure evaluation of humans who have ingested diacylglycerol oil containing glycidol fatty acid esters using hemoglobin adducts.

    PubMed

    Honda, Hiroshi; Fujii, Kenkichi; Yamaguchi, Tohru; Ikeda, Naohiro; Nishiyama, Naohiro; Kasamatsu, Toshio

    2012-11-01

    Glycidol fatty acid esters (GEs) have been found as impurities in refined edible oils including diacylglycerol (DAG) oil, and concerns of possible exposure to glycidol (G), a known animal carcinogen, during digestion have been raised. We previously measured N-(2,3-dihydroxy-propyl)valine (diHOPrVal), a G hemoglobin adduct, for DAG oil exposed and non-exposed groups and showed there was no significant difference between them. In the present study, we conducted an additional analysis to verify the outcome of the previous report. The first experiment was designed as a matched case-control study to adjust variables with an increased sample size. The average levels of diHOPrVal were 6.9 pmol/g-globin (95%CI: 4.9-9.0) for 14 DAG oil exposed subjects and 7.3 pmol/g-globin (95%CI: 6.1-8.5) for 42 non-exposed volunteers, and no significant difference in levels was found between the two groups. In a second experiment, we compared the adduct levels of 12 DAG oil exposed subjects before and after discontinuing use of DAG oil, and found there was no significant change in diHOPrVal levels (from 7.1±1.1 to 7.5±1.4 pmol/g-globin). These results suggest that there was no increased exposure to G for humans who ingested DAG oil daily, although the evaluated population was limited. PMID:22889899

  16. Does Diacylglycerol Accumulation in Fatty Liver Disease Cause Hepatic Insulin Resistance?

    PubMed Central

    Finck, Brian N.; Hall, Angela M.

    2015-01-01

    Numerous studies conducted on obese humans and various rodent models of obesity have identified a correlation between hepatic lipid content and the development of insulin resistance in liver and other tissues. Despite a large body of the literature on this topic, the cause and effect relationship between hepatic steatosis and insulin resistance remains controversial. If, as many believe, lipid aggregation in liver drives insulin resistance and other metabolic abnormalities, there are significant unanswered questions as to which lipid mediators are causative in this cascade. Several published papers have now correlated levels of diacylglycerol (DAG), the penultimate intermediate in triglyceride synthesis, with development of insulin resistance and have postulated that this occurs via activation of protein kinase C signaling. Although many studies have confirmed this relationship, many others have reported a disconnect between DAG content and insulin resistance. It has been postulated that differences in methods for DAG measurement, DAG compartmentalization within the cell, or fatty acid composition of the DAG may explain these discrepancies. The purpose of this review is to compare and contrast some of the relevant findings in this area and to discuss a number of unanswered questions regarding the relationship between DAG and insulin resistance. PMID:26273583

  17. Tracking Diacylglycerol and Phosphatidic Acid Pools in Budding Yeast

    PubMed Central

    Ganesan, Suriakarthiga; Shabits, Brittney N.; Zaremberg, Vanina

    2015-01-01

    Phosphatidic acid (PA) and diacylglycerol (DAG) are key signaling molecules and important precursors for the biosynthesis of all glycerolipids found in eukaryotes. Research conducted in the model organism Saccharomyces cerevisiae has been at the forefront of the identification of the enzymes involved in the metabolism and transport of PA and DAG. Both these lipids can alter the local physical properties of membranes by introducing negative curvature, but the anionic nature of the phosphomonoester headgroup in PA sets it apart from DAG. As a result, the mechanisms underlying PA and DAG interaction with other lipids and proteins are notoriously different. This is apparent from the analysis of the protein domains responsible for recognition and binding to each of these lipids. We review the current evidence obtained using the PA-binding proteins and domains fused to fluorescent proteins for in vivo tracking of PA pools in yeast. In addition, we present original results for visualization of DAG pools in yeast using the C1 domain from mammalian PKCδ. An emerging first cellular map of the distribution of PA and DAG pools in actively growing yeast is discussed. PMID:27081314

  18. Regulation of canonical transient receptor potential (TRPC) channel function by diacylglycerol and protein kinase C.

    PubMed

    Venkatachalam, Kartik; Zheng, Fei; Gill, Donald L

    2003-08-01

    The mechanism of receptor-induced activation of the ubiquitously expressed family of mammalian canonical transient receptor potential (TRPC) channels has been the focus of intense study. Primarily responding to phospholipase C (PLC)-coupled receptors, the channels are reported to receive modulatory input from diacylglycerol, endoplasmic reticulum inositol 1,4,5-trisphosphate receptors and Ca2+ stores. Analysis of TRPC5 channels transfected within DT40 B cells and deletion mutants thereof revealed efficient activation in response to PLC-beta or PLC-gamma activation, which was independent of inositol 1,4,5-trisphoshate receptors or the content of stores. In both HEK293 cells and DT40 cells, TRPC5 and TRPC3 channel responses to PLC activation were highly analogous, but only TRPC3 and not TRPC5 channels responded to the addition of the permeant diacylglycerol (DAG) analogue, 1-oleoyl-2-acetyl-sn-glycerol (OAG). However, OAG application or elevated endogenous DAG, resulting from either DAG lipase or DAG kinase inhibition, completely prevented TRPC5 or TRPC4 activation. This inhibitory action of DAG on TRPC5 and TRPC4 channels was clearly mediated by protein kinase C (PKC), in distinction to the stimulatory action of DAG on TRPC3, which is established to be PKC-independent. PKC activation totally blocked TRPC3 channel activation in response to OAG, and the activation was restored by PKC-blockade. PKC inhibition resulted in decreased TRPC3 channel deactivation. Store-operated Ca2+ entry in response to PLC-coupled receptor activation was substantially reduced by OAG or DAG-lipase inhibition in a PKC-dependent manner. However, store-operated Ca2+ entry in response to the pump blocker, thapsigargin, was unaffected by PKC. The results reveal that each TRPC subtype is strongly inhibited by DAG-induced PKC activation, reflecting a likely universal feedback control on TRPCs, and that DAG-mediated PKC-independent activation of TRPC channels is highly subtype-specific. The

  19. Experimental study of non-linear effects in a typical shear lap joint configuration

    NASA Astrophysics Data System (ADS)

    Hartwigsen, C. J.; Song, Y.; McFarland, D. M.; Bergman, L. A.; Vakakis, A. F.

    2004-10-01

    Although mechanical joints are integral parts of most practical structures, their modelling and their effects on structural dynamics are not yet fully understood. This represents a serious impediment to accurate modelling of the dynamics and to the development of reduced-order, finite element models capable of describing the effects of mechanical joints on the dynamics. In this work we provide an experimental study to quantify the non-linear effects of a typical shear lap joint on the dynamics of two structures: a beam with a bolted joint in its center; and a frame with a bolted joint in one of its members. Both structures are subjected to a variety of dynamical tests to determine the non-linear effects of the joints. The tests reveal several important influences on the effective stiffness and damping of the lap joints. The possibility of using Iwan models to represent the experimentally observed joint effects is discussed.

  20. A two-sided interface model for dissipation in structural systems with frictional joints

    NASA Astrophysics Data System (ADS)

    Miller, Jason D.; Dane Quinn, D.

    2009-03-01

    Modeling mechanical joints in an accurate and computationally efficient manner is of great importance in the analysis of structural systems, which can be composed of a large number of connected components. This work presents an interface model that can be decomposed into a series-series Iwan model together with an elastic chain, subject to interfacial shear loads. A reduced-order formulation of the resulting model is developed that significantly reduces the computational requirements for the simulation of frictional damping. Results are presented as the interface is subject to harmonic loading of varying amplitude. The models presented are able to qualitatively reproduce experimentally observed dissipation scalings. Finally, the interface models are embedded within a larger structural system to illustrate there effectiveness in capturing the structural damping induced by mechanical joints.

  1. A two-state hysteresis model from high-dimensional friction

    PubMed Central

    Biswas, Saurabh; Chatterjee, Anindya

    2015-01-01

    In prior work (Biswas & Chatterjee 2014 Proc. R. Soc. A 470, 20130817 (doi:10.1098/rspa.2013.0817)), we developed a six-state hysteresis model from a high-dimensional frictional system. Here, we use a more intuitively appealing frictional system that resembles one studied earlier by Iwan. The basis functions now have simple analytical description. The number of states required decreases further, from six to the theoretical minimum of two. The number of fitted parameters is reduced by an order of magnitude, to just six. An explicit and faster numerical solution method is developed. Parameter fitting to match different specified hysteresis loops is demonstrated. In summary, a new two-state model of hysteresis is presented that is ready for practical implementation. Essential Matlab code is provided. PMID:26587279

  2. A two-state hysteresis model from high-dimensional friction.

    PubMed

    Biswas, Saurabh; Chatterjee, Anindya

    2015-07-01

    In prior work (Biswas & Chatterjee 2014 Proc. R. Soc. A 470, 20130817 (doi:10.1098/rspa.2013.0817)), we developed a six-state hysteresis model from a high-dimensional frictional system. Here, we use a more intuitively appealing frictional system that resembles one studied earlier by Iwan. The basis functions now have simple analytical description. The number of states required decreases further, from six to the theoretical minimum of two. The number of fitted parameters is reduced by an order of magnitude, to just six. An explicit and faster numerical solution method is developed. Parameter fitting to match different specified hysteresis loops is demonstrated. In summary, a new two-state model of hysteresis is presented that is ready for practical implementation. Essential Matlab code is provided. PMID:26587279

  3. Structural Elucidation of Diglycosyl Diacylglycerol and Monoglycosyl Diacylglycerol from Streptococcus pneumoniae by Multiple-Stage Linear Ion-Trap Mass Spectrometry with Electrospray Ionization

    PubMed Central

    Tatituri, Raju Venkata Veera; Brenner, Michael B.; Turk, John; Hsu, Fong-Fu

    2013-01-01

    The cell wall of the pathogenic bacterium Streptococcus pneumoniae (S. pneumoniae) contains glucopyranosyl diacylglycerol (GlcDAG) and galactoglucopyranosyldiacylglycerol (GalGlcDAG). The specific GlcDAG consisting of vaccenic acid substituent at sn-2 was recently identified as another glycolipid antigen family recognized by invariant natural killer T cells (iNKT cells). Here, we describe a linear ion-trap (LIT) multiple-stage (MSn) mass spectrometric approach towards structural analysis of GalGlcDAG and GlcDAG. Structural information derived from MSn (n = 2,3) on the [M + Li]+ adduct ions desorbed by electrospray ionization (ESI) affords identification of the fatty acid substituents, assignment of the fatty acyl groups on the glycerol backbone, as well as the location of double bond along the fatty acyl chain. The identification of the fatty acyl groups and determination of their regio-specificity were confirmed by MSn (n = 2,3) on the [M + NH4]+ ions. We establish the structures of GalGlcDAG and GlcDAG isolated from S. pneumoniae, in which the major species consists of a 16:1- or 18:1-fatty acid substituent mainly at sn-2, and the double bond of the fatty acid is located at ω-7 (n-7). More than one isomers were found for each mass in the family. This mass spectrometric approach provides a simple method to achieve structure identification of this important lipid family that would be very difficult to define using the traditional method. PMID:22282097

  4. Interfacial partitioning of a loop hinge residue contributes to diacylglycerol affinity of conserved region 1 domains.

    PubMed

    Stewart, Mikaela D; Cole, Taylor R; Igumenova, Tatyana I

    2014-10-01

    Conventional and novel isoenzymes of PKC are activated by the membrane-embedded second messenger diacylglycerol (DAG) through its interactions with the C1 regulatory domain. The affinity of C1 domains to DAG varies considerably among PKCs. To gain insight into the origin of differential DAG affinities, we conducted high-resolution NMR studies of C1B domain from PKCδ (C1Bδ) and its W252Y variant. The W252Y mutation was previously shown to render C1Bδ less responsive to DAG (Dries, D. R., Gallegos, L. L., and Newton, A. C. (2007) A single residue in the C1 domain sensitizes novel protein kinase C isoforms to cellular diacylglycerol production. J. Biol. Chem. 282, 826-830) and thereby emulate the behavior of C1B domains from conventional PKCs that have a conserved Tyr at the equivalent position. Our data revealed that W252Y mutation did not perturb the conformation of C1Bδ in solution but significantly reduced its propensity to partition into a membrane-mimicking environment in the absence of DAG. Using detergent micelles doped with a paramagnetic lipid, we determined that both the residue identity at position 252 and complexation with diacylglycerol influence the geometry of C1Bδ-micelle interactions. In addition, we identified the C-terminal helix α1 of C1Bδ as an interaction site with the head groups of phosphatidylserine, a known activator of PKCδ. Taken together, our studies (i) reveal the identities of C1Bδ residues involved in interactions with membrane-mimicking environment, DAG, and phosphatidylserine, as well as the affinities associated with each event and (ii) suggest that the initial ligand-independent membrane recruitment of C1B domains, which is greatly facilitated by the interfacial partitioning of Trp-252, is responsible, at least in part, for the differential DAG affinities. PMID:25124034

  5. Interfacial Partitioning of a Loop Hinge Residue Contributes to Diacylglycerol Affinity of Conserved Region 1 Domains*

    PubMed Central

    Stewart, Mikaela D.; Cole, Taylor R.; Igumenova, Tatyana I.

    2014-01-01

    Conventional and novel isoenzymes of PKC are activated by the membrane-embedded second messenger diacylglycerol (DAG) through its interactions with the C1 regulatory domain. The affinity of C1 domains to DAG varies considerably among PKCs. To gain insight into the origin of differential DAG affinities, we conducted high-resolution NMR studies of C1B domain from PKCδ (C1Bδ) and its W252Y variant. The W252Y mutation was previously shown to render C1Bδ less responsive to DAG (Dries, D. R., Gallegos, L. L., and Newton, A. C. (2007) A single residue in the C1 domain sensitizes novel protein kinase C isoforms to cellular diacylglycerol production. J. Biol. Chem. 282, 826–830) and thereby emulate the behavior of C1B domains from conventional PKCs that have a conserved Tyr at the equivalent position. Our data revealed that W252Y mutation did not perturb the conformation of C1Bδ in solution but significantly reduced its propensity to partition into a membrane-mimicking environment in the absence of DAG. Using detergent micelles doped with a paramagnetic lipid, we determined that both the residue identity at position 252 and complexation with diacylglycerol influence the geometry of C1Bδ-micelle interactions. In addition, we identified the C-terminal helix α1 of C1Bδ as an interaction site with the head groups of phosphatidylserine, a known activator of PKCδ. Taken together, our studies (i) reveal the identities of C1Bδ residues involved in interactions with membrane-mimicking environment, DAG, and phosphatidylserine, as well as the affinities associated with each event and (ii) suggest that the initial ligand-independent membrane recruitment of C1B domains, which is greatly facilitated by the interfacial partitioning of Trp-252, is responsible, at least in part, for the differential DAG affinities. PMID:25124034

  6. A study of the influence of charged residues on β-hairpin formation by nuclear magnetic resonance and molecular dynamics.

    PubMed

    Makowska, Joanna; Zmudzińska, Wioletta; Uber, Dorota; Chmurzyński, Lech

    2014-12-01

    Chain reversals are often nucleation sites in protein folding. The β-hairpins of FBP28 WW domain and IgG are stable and have been proved to initiate the folding and are, therefore, suitable for studying the influence of charged residues on β-hairpin conformation. In this paper, we carried out NMR examination of the conformations in solution of two fragments from the FPB28 protein (PDB code: 1E0L) (N-terminal part) namely KTADGKT-NH2 (1E0L 12-18, D7) and YKTADGKTY-NH2 (1E0L 11-19, D9), one from the B3 domain of the protein G (PDB code: 1IGD), namely DDATKT-NH2 (1IGD 51-56) (Dag1), and three variants of Dag1 peptide: DVATKT-NH2 (Dag2), OVATKT-NH2 (Dag3) and KVATKT-NH2 (Dag4), respectively, in which the original charged residue were replaced with non-polar residues or modified charged residues. It was found that both the D7 and D9 peptides form a large fraction bent conformations. However, no hydrophobic contacts between the terminal Tyr residues of D9 occur, which suggests that the presence of a pair of like-charged residues stabilizes chain reversal. Conversely, only the Dag1 and Dag2 peptides exhibit some chain reversal; replacing the second aspartic-acid residue with a valine and the first one with a basic residue results in a nearly extended conformation. These results suggest that basic residues farther away in sequence can result in stabilization of chain reversal owing to screening of the non-polar core. Conversely, smaller distance in sequence prohibits this screening, while the presence oppositely-charged residues can stabilize a turn because of salt-bridge formation. PMID:25316116

  7. Heterozygous deletion of a 2-Mb region including the dystroglycan gene in a patient with mild myopathy, facial hypotonia, oral-motor dyspraxia and white matter abnormalities.

    PubMed

    Frost, Amy R; Böhm, Sabrina V; Sewduth, Raj N; Josifova, Dragana; Ogilvie, Caroline Mackie; Izatt, Louise; Roberts, Roland G

    2010-07-01

    Dystroglycan is a protein which binds directly to two proteins defective in muscular dystrophies (dystrophin and laminin alpha2) and whose own aberrant post-translational modification is the common aetiological route of neuromuscular diseases associated with mutations in genes encoding at least six other proteins (POMT1, POMT2, POMGnT1, LARGE, FKTN and FKRP). It is surprising, therefore, that to our knowledge no mutations of the human dystroglycan gene itself have yet been reported. In this study, we describe a patient with a heterozygous de novo deletion of a approximately 2-Mb region of chromosome 3, which includes the dystroglycan gene (DAG1). The patient is a 16-year-old female with learning difficulties, white matter abnormalities, elevated serum creatine kinase, oral-motor dyspraxia and facial hypotonia but minimal clinically significant involvement of other muscles. As these symptoms are a subset of those observed in disorders of dystroglycan glycosylation (muscle-eye-brain disease and Warker-Warburg syndrome), we assess the likely contribution to her phenotype of her heterogosity for a null mutation of DAG1. We also show that the transcriptional compensation observed in the Dag1(+/-) mouse is not observed in the patient. Although we cannot show that haploinsufficiency of DAG1 is the sole cause of this patient's myopathy and white matter changes, this case serves to constrain our ideas of the severity of the phenotypic consequences of heterozygosity for null DAG1 mutations. PMID:20234391

  8. Heterozygous deletion of a 2-Mb region including the dystroglycan gene in a patient with mild myopathy, facial hypotonia, oral-motor dyspraxia and white matter abnormalities

    PubMed Central

    Frost, Amy R; Böhm, Sabrina V; Sewduth, Raj N; Josifova, Dragana; Ogilvie, Caroline Mackie; Izatt, Louise; Roberts, Roland G

    2010-01-01

    Dystroglycan is a protein which binds directly to two proteins defective in muscular dystrophies (dystrophin and laminin α2) and whose own aberrant post-translational modification is the common aetiological route of neuromuscular diseases associated with mutations in genes encoding at least six other proteins (POMT1, POMT2, POMGnT1, LARGE, FKTN and FKRP). It is surprising, therefore, that to our knowledge no mutations of the human dystroglycan gene itself have yet been reported. In this study, we describe a patient with a heterozygous de novo deletion of a ∼2-Mb region of chromosome 3, which includes the dystroglycan gene (DAG1). The patient is a 16-year-old female with learning difficulties, white matter abnormalities, elevated serum creatine kinase, oral-motor dyspraxia and facial hypotonia but minimal clinically significant involvement of other muscles. As these symptoms are a subset of those observed in disorders of dystroglycan glycosylation (muscle–eye–brain disease and Warker–Warburg syndrome), we assess the likely contribution to her phenotype of her heterogosity for a null mutation of DAG1. We also show that the transcriptional compensation observed in the Dag1+/− mouse is not observed in the patient. Although we cannot show that haploinsufficiency of DAG1 is the sole cause of this patient's myopathy and white matter changes, this case serves to constrain our ideas of the severity of the phenotypic consequences of heterozygosity for null DAG1 mutations. PMID:20234391

  9. Diacylglycerol kinase-ζ regulates mTORC1 and lipogenic metabolism in cancer cells through SREBP-1

    PubMed Central

    Torres-Ayuso, P; Tello-Lafoz, M; Mérida, I; Ávila-Flores, A

    2015-01-01

    Diacylglycerol kinases (DGKs) transform diacylglycerol (DAG) into phosphatidic acid (PA), balancing the levels of these key metabolic and signaling lipids. We previously showed that PA derived from the DGKζ isoform promotes mammalian target of rapamycin complex 1 (mTORC1) activation. This function might be crucial for the growth and survival of cancer cells, especially for those resistant to the allosteric mTOR inhibitor rapamycin. How this positive function of DGKζ coordinates with DAG metabolism and signaling is unknown. In this study, we used a rapamycin-resistant colon cancer cell line as a model to address the role of DGKζ in tumor cells. We found that DGKζ predominated over other PA sources such as DGKα or phospholipase D to activate mTORC1, and that its activity was a component of the rapamycin-induced feedback loops. We show that the DGKζ DAG-consuming function is central to cell homeostasis, as DAG negatively regulates levels of the lipogenic transcription factor SREBP-1. Our findings suggest a model in which simultaneous regulation of DAG and PA levels by DGKζ is integrated with mTOR function to maintain tumor cell homeostasis; we provide new evidence of the crosstalk between mTOR and lipid metabolism that will be advantageous in the design of drug therapies. PMID:26302180

  10. Differential effects of pertussis toxin on insulin-stimulated phosphatidylcholine hydrolysis and glycerolipid synthesis de novo. Studies in BC3H-1 myocytes and rat adipocytes

    SciTech Connect

    Hoffman, J.M.; Standaert, M.L.; Nair, G.P.; Farese, R.V. )

    1991-04-02

    Insulin-induced increases in diacylglycerol (DAG) have been suggested to result from stimulation of de novo phosphatidic acid (PA) synthesis and phosphatidylcholine (PC) hydrolysis. Presently, the authors found that insulin decreased PC levels of BC3H-1 myocytes and rat adipocytes by approximately 10-25% within 30 s. These decreases were rapidly reversed in both cell types, apparently because of increased PC synthesis de novo. In BC3H-1 myocytes, pertussis toxin inhibited PC resynthesis and insulin effects on the pathway of de novo PA-DAG-PC synthesis, as evidenced by changes in ({sup 3}H)glycerol incorporation, but did not inhibit insulin-stimulated PC hydrolysis. Pertussis toxin also blocked the later, but not the initial, increase in DAG production in the myocytes. Phorbol esters activated PC hydrolysis in both myocytes and adipocytes, but insulin-induced stimulation of PC hydrolysis was not dependent upon activation of PKC, since this hydrolysis was not inhibited by 500 {mu}M sangivamycin, an effective PKC inhibitor. The results indicate that insulin increases DAG by pertussis toxin sensitive and insensitive (PC hydrolysis) mechanisms, which are mechanistically separate, but functionally interdependent and integrated. PC hydrolysis may contribute importantly to initial increases in DAG, but later sustained increases are apparently largely dependent on insulin-induced stimulation of the pathway of de novo phospholipid synthesis.

  11. Non-specific phospholipase C5 and diacylglycerol promote lateral root development under mild salt stress in Arabidopsis.

    PubMed

    Peters, Carlotta; Kim, Sang-Chul; Devaiah, Shivakumar; Li, Maoyin; Wang, Xuemin

    2014-09-01

    Developing a robust root system is crucial to plant survival and competition for soil resources. Here we report that the non-specific phospholipase C5 (NPC5) and its derived lipid mediator diacylglycerol (DAG) mediate lateral root (LR) development during salt stress in Arabidopsis thaliana. T-DNA knockout mutant npc5-1 produced few to no LR under mild NaCl stress, whereas overexpression of NPC5 increased LR number. Roots of npc5-1 contained a lower level of DAG than wild type, whereas NPC5 overexpressor exhibited an increase in DAG level. Application of DAG, but not phosphatidic acid, fully restored LR growth of npc5-1 to that of wild type under NaCl stress. NPC5 expression was significantly induced in Arabidopsis seedlings treated with NaCl. Npc5-1 was less responsive to auxin-mediated root growth than the wild type. These results indicate that NPC5 mediates LR development in response to salt stress and suggest that DAG functions as a lipid mediator in the stress signalling. PMID:24689655

  12. Munc13-1-mediated vesicle priming contributes to secretory amyloid precursor protein processing.

    PubMed

    Rossner, Steffen; Fuchsbrunner, Katrin; Lange-Dohna, Christine; Hartlage-Rübsamen, Maike; Bigl, Volker; Betz, Andrea; Reim, Kerstin; Brose, Nils

    2004-07-01

    The amyloid precursor protein (APP) gives rise toc beta-amyloid peptides, which are the main constituents of senile plaques in brains of Alzheimer's disease patients. Non-amyloidogenic processing of the APP can be stimulated by phorbol esters (PEs) and by intracellular diacylglycerol (DAG) generation. This led to the hypothesis that classical and novel protein kinase Cs (PKCs), which are activated by DAG/PEs, regulate APP processing. However, in addition to PKCs, there are other DAG/PE receptors present in neurons that may participate in the modulation of APP processing. Munc13-1, a presynaptic protein with an essential role in synaptic vesicle priming, represents such an alternative target of the DAG second messenger pathway. Using Munc13-1 knock-out mice and knock-in mice expressing a Munc13-1(H567K) variant deficient in DAG/PE binding, we determined the relative contributions of PKCs and Munc13-1 to PE-stimulated secretory APP processing. We establish that, in addition to PKC, Munc13-1 significantly contributes to the regulation of secretory APP metabolism. PMID:15123597

  13. Diacylglycerol kinase ζ limits B cell antigen receptor-dependent activation of ERK signaling to inhibit early antibody responses.

    PubMed

    Wheeler, Matthew L; Dong, Matthew B; Brink, Robert; Zhong, Xiao-Ping; DeFranco, Anthony L

    2013-10-15

    Signaling downstream of the B cell antigen receptor (BCR) is tightly regulated to enable cells to gauge the strength and duration of antigen-receptor interactions and to respond appropriately. We investigated whether metabolism of the second messenger diacylglycerol (DAG) by members of the family of DAG kinases (DGKs) played a role in modulating the magnitude of signaling by DAG downstream of the BCR. In the absence of DGKζ, the threshold for BCR signaling, measured as activation of the Ras-extracellular signal-regulated kinase (ERK) pathway, was markedly reduced in mature follicular B cells, which resulted in enhanced responses to antigen in vitro and in vivo. Inhibition of DAG signaling by DGKζ limited the number of antibody-secreting cells that were generated early in response to T cell-independent type 2 antigens, as well as to T cell-dependent antigens. Furthermore, the effect of loss of DGKζ closely resembled the effect of increasing the affinity of the BCR for antigen during the T cell-dependent antibody response. These results suggest that the magnitude of DAG signaling is important for translating the affinity of the BCR for antigen into the amount of antibody produced during the early stages of an immune response. PMID:24129701

  14. [How independent pharmacological screenings in plants and humans led to the discovery of a new family of lipid metabolism inhibitors].

    PubMed

    Chevalier, Florian; Maréchal, Éric

    2015-03-01

    In eukaryotic cells, phosphatidic acid (PA) and diacylglycerol (DAG), are at the origin of all membrane glycerolipids. Their interconversion is achieved by dephosphorylation of PA and phosphorylation of DAG: they form therefore a metabolic hub. PA and DAG are also known to be versatile signaling molecules. Two independent pharmacological screenings conducted on plant and human targets, led to the discovery of a new family of compounds acting on enzymes binding to either PA or DAG, in biological contexts that seemed initially independent. On the one hand, in plants, monogalactosyldiacylglycerol synthases (MGDG synthases or MGD) are responsible for the synthesis of MGDG, which is the most profuse lipid of photosynthetic membranes, and thus essential for metabolism and development. MGD use DAG as substrate. On the other hand, in mammals, phospholipases D (PLD), that produce PA, are involved in a variety of signaling cascades that control a broad spectrum of cellular functions, and play a role in the development of cancers. The two independent pharmacological screenings described in this review aimed to identify inhibitory molecules of either MGD of the plant model Arabidopsis, or human PLD. In both cases, the obtained molecules are piperidinyl-benzimidazolone derivatives, thereby allowing to propose this family of molecules as a novel source of inspiration for the search of compounds interfering with glycerolipid metabolism, that could be useful for other biological and therapeutics contexts. PMID:25855286

  15. Assaying nonspecific phospholipase C activity.

    PubMed

    Pejchar, Přemysl; Scherer, Günther F E; Martinec, Jan

    2013-01-01

    Plant nonspecific phospholipase C (NPC) is a recently described enzyme which plays a role in membrane rearrangement during phosphate starvation. It is also involved in responses of plants to brassinolide, abscisic acid (ABA), elicitors, and salt. The NPC activity is decreased in cells treated with aluminum. In the case of salt stress, the molecular mechanism of NPC action is based on accumulation of diacylglycerol (DAG) by hydrolysis of phospholipids and conversion of DAG, the product of NPC activity, to phosphatidic acid (PA) that participates in ABA signaling pathways. Here we describe a step-by-step protocol, which can be used to determine in situ or in vitro NPC activity. Determination is based on quantification of fluorescently labeled DAG as a product of cleavage of the fluorescently labeled substrate lipid, phosphatidylcholine. High-performance thin-layer chromatography is used for separation of fluorescent DAG. The spot is visualized with a laser scanner and the relative amounts of fluorescent DAG are quantified using imaging software. PMID:23681535

  16. Distinctive roles of PLD signaling elicited by oxidative stress in synaptic endings from adult and aged rats.

    PubMed

    Mateos, Melina V; Giusto, Norma M; Salvador, Gabriela A

    2012-12-01

    The role of iron in oxidative injury in the nervous system has been extensively described. However, little is known about the role of lipid signal transduction in neurodegeneration processes triggered by iron overload. The purpose of this work was to characterize the regulation and the crosstalk between phosphatidylcholine (PC)-derived diacylglycerol (DAG) and cannonical signaling pathways during iron-induced oxidative stress in cerebral cortex synaptic endings (Syn) obtained from adult (4 months old) and aged (28 months old) rats. DAG production was increased in Syn exposed to iron. This rise in DAG formation was due to phospholipase D1 (PLD1) and PLD2 activations. In adult rats, PKD1, ERK1/2 and PKCα/βII activations were PLD1 and PLD2 dependent. In contrast, in senile rats, DAG formation catalyzed by PLDs did not participate in PKD1, ERK1/2 and PKCα/βII regulations, but it was dependent on ERK and PKC activities. Iron-induced oxidative stress promoted an increased localization of PLD1 in membrane rafts, whereas PLD2 was excluded from these domains and appeared to be involved in glutamate transporter function. Our results show a differential regulation and synaptic function of DAG generated by PLDs during iron-induced oxidative stress as a consequence of aging. PMID:23010583

  17. Physicochemical properties of peanut oil-based diacylglycerol and their derived oil-in-water emulsions stabilized by sodium caseinate.

    PubMed

    Long, Zhao; Zhao, Mouming; Liu, Ning; Liu, Daolin; Sun-Waterhouse, Dongxiao; Zhao, Qiangzhong

    2015-10-01

    High purity peanut oil-based diacylglycerol (PO-DAG) (94.95 wt%) was prepared via enzymatic glycerolysis from peanut oil (PO). The resulting dominance of DAGs was proven to greatly influence the properties of corresponding fresh or frozen-thawed emulsions. Stable fresh oil-in-water emulsions were produced using either PO-DAG or PO, with stability enhanced by increased concentrations of Na-CN. The lower equilibrium interfacial tension along with greater negative ζ-potential of PO revealed that Na-CN was preferentially adsorbed to the PO interface. Adding 0.05 mol/L NaCl to the PO emulsions minimized depletion flocculation caused by the unadsorbed Na-CN, but further NaCl addition increased oil droplet size and concomitant coalescence. For the PO-DAG emulsions, adding 0.2 mol/L NaCl did not significantly (p>0.05) affect their ζ-potential but adding 0.05 or 0.1 mol/L NaCl lowered ζ-potential, although NaCl at these concentrations increased oil droplet size and coalescence. Freezing-thawing process considerably weakened the stability of PO-DAG emulsions. PMID:25872432

  18. Fact Sheets of CTAS and NASA Decision-Support Tools and Concepts

    NASA Technical Reports Server (NTRS)

    Lee, Katharine

    2004-01-01

    Distributed Air/Ground (DAG) Traffic Management (TM) is an integrated operational concept in which flight deck crews, air traffic service providers and aeronautical operational control personnel use distributed decision-making to enable user preferences and increase system capacity, while meeting air traffic management (ATM) requirements. It is a possible operational mode under the Free Flight concept outlined by the RTCA Task Force 3. The goal of DAG-TM is to enhance user flexibility/efficiency and increase system capacity, without adversely affecting system safety or restricting user accessibility to the National Airspace System (NAS). DAG-TM will be accomplished with a human-centered operational paradigm enabled by procedural and technological innovations. These innovations include automation aids, information sharing and Communication, Navigation, and Surveillance (CNS) / ATM technologies. The DAG-TM concept is intended to eliminate static restrictions to the maximum extent possible. In this paradigm, users may plan and operate according to their preferences - as the rule rather than the exception - with deviations occumng eyond the year 2015. Out of a total of 15 concept elements, 4 have been selected for initial sutidies (see Key Elements in sidebar). DAG-TM research is being performed at Ames, Glenn, and Langley Research Centers.

  19. A Directed Acyclic Graph-Large Margin Distribution Machine Model for Music Symbol Classification

    PubMed Central

    Wen, Cuihong; Zhang, Jing; Rebelo, Ana; Cheng, Fanyong

    2016-01-01

    Optical Music Recognition (OMR) has received increasing attention in recent years. In this paper, we propose a classifier based on a new method named Directed Acyclic Graph-Large margin Distribution Machine (DAG-LDM). The DAG-LDM is an improvement of the Large margin Distribution Machine (LDM), which is a binary classifier that optimizes the margin distribution by maximizing the margin mean and minimizing the margin variance simultaneously. We modify the LDM to the DAG-LDM to solve the multi-class music symbol classification problem. Tests are conducted on more than 10000 music symbol images, obtained from handwritten and printed images of music scores. The proposed method provides superior classification capability and achieves much higher classification accuracy than the state-of-the-art algorithms such as Support Vector Machines (SVMs) and Neural Networks (NNs). PMID:26985826

  20. Multi-Resolution Dynamic Meshes with Arbitrary Deformations

    SciTech Connect

    Shamir, A.; Pascucci, V.; Bajaj, C.

    2000-07-10

    Multi-resolution techniques and models have been shown to be effective for the display and transmission of large static geometric object. Dynamic environments with internally deforming models and scientific simulations using dynamic meshes pose greater challenges in terms of time and space, and need the development of similar solutions. In this paper we introduce the T-DAG, an adaptive multi-resolution representation for dynamic meshes with arbitrary deformations including attribute, position, connectivity and topology changes. T-DAG stands for Time-dependent Directed Acyclic Graph which defines the structure supporting this representation. We also provide an incremental algorithm (in time) for constructing the T-DAG representation of a given input mesh. This enables the traversal and use of the multi-resolution dynamic model for partial playback while still constructing new time-steps.

  1. A Directed Acyclic Graph-Large Margin Distribution Machine Model for Music Symbol Classification.

    PubMed

    Wen, Cuihong; Zhang, Jing; Rebelo, Ana; Cheng, Fanyong

    2016-01-01

    Optical Music Recognition (OMR) has received increasing attention in recent years. In this paper, we propose a classifier based on a new method named Directed Acyclic Graph-Large margin Distribution Machine (DAG-LDM). The DAG-LDM is an improvement of the Large margin Distribution Machine (LDM), which is a binary classifier that optimizes the margin distribution by maximizing the margin mean and minimizing the margin variance simultaneously. We modify the LDM to the DAG-LDM to solve the multi-class music symbol classification problem. Tests are conducted on more than 10000 music symbol images, obtained from handwritten and printed images of music scores. The proposed method provides superior classification capability and achieves much higher classification accuracy than the state-of-the-art algorithms such as Support Vector Machines (SVMs) and Neural Networks (NNs). PMID:26985826

  2. Analysis of the Staphylococcus aureus DgkB Structure Reveals a Common Catalytic Mechanism for the Soluble Diacylglycerol Kinases

    SciTech Connect

    Miller, Darcie J.; Jerga, Agoston; Rock, Charles O.; White, Stephen W.

    2008-08-11

    Soluble diacylglycerol (DAG) kinases function as regulators of diacylglycerol metabolism in cell signaling and intermediary metabolism. We report the structure of a DAG kinase, DgkB from Staphylococcus aureus, both as the free enzyme and in complex with ADP. The molecule is a tight homodimer, and each monomer comprises two domains with the catalytic center located within the interdomain cleft. Two distinctive features of DkgB are a structural Mg{sup 2+} site and an associated Asp{center_dot}water{center_dot}Mg{sup 2+} network that extends toward the active site locale. Site-directed mutagenesis revealed that these features play important roles in the catalytic mechanism. The key active site residues and the components of the Asp{center_dot}water{center_dot}Mg{sup 2+} network are conserved in the catalytic cores of the mammalian signaling DAG kinases, indicating that these enzymes use the same mechanism and have similar structures as DgkB.

  3. The click-compatible sugar 6-deoxy-alkynyl glucose metabolically incorporates into Arabidopsis root hair tips and arrests their growth.

    PubMed

    McClosky, Daniel D; Wang, Bo; Chen, Gong; Anderson, Charles T

    2016-03-01

    Plant cell walls are dynamic structures whose polysaccharide components are rearranged and recycled during growth and morphogenesis. Covalent fluorescent tagging of these polysaccharides following a metabolic labeling approach can help elucidate these changes. Herein reported are the synthesis and seedling-incorporation of a plant polysaccharide chemical reporter, 6-deoxy-alkynyl glucose (6dAG), that is modeled on D-glucose. Whereas fucose-alkyne, a previously reported chemical reporter for pectin, incorporates diffusely throughout growing cell walls, 6dAG incorporated specifically into root hair tips. This incorporation occurs in a time- and concentration-dependent manner. 6dAG exposure both induces and colocalizes with callose deposition in this tissue, and arrests both root hair and root growth. These results show that plants can incorporate an additional alkynyl-modified sugar analog into their metabolism, and into a discrete subcellular location. PMID:26833385

  4. Protein kinase C activity in the spleen of trout (Salmo gairdneri) and the rectal gland of dogfish (Scyliorhinus canicula), and the effects of phosphatidylserine and diacylglycerol containing (n-3) polyunsaturated fatty acids.

    PubMed

    Bell, M V; Sargent, J R

    1987-01-01

    1. High speed supernatant fractions of trout spleen and dogfish rectal gland contained 22.5 and 7.2 nmol/min/g tissue of protein kinase C activity respectively. 2. The effect of Ca2+ concentration on the activities with phosphatidylserine (PtdSer) alone, diacylglycerol (DAG) alone and PtdSer and DAG together were determined. Both enzymes required Ca2+ but activity was independent of Ca2+ concentration within the physiological range of 0.1-10 microns. 3. The effect of PtdSer and DAG containing (n - 3) polyunsaturated fatty acids (PUFA) on the activity of protein kinase C from both tissues was examined. Both enzymes were active with all lipids tested and showed little or no discrimination between lipids differing in their contents of (n-3) or (n-6) polyunsaturated fatty acids. PMID:3665435

  5. Clostridium perfringens Alpha-Toxin Induces Gm1a Clustering and Trka Phosphorylation in the Host Cell Membrane.

    PubMed

    Takagishi, Teruhisa; Oda, Masataka; Kabura, Michiko; Kurosawa, Mie; Tominaga, Kaori; Urano, Shiori; Ueda, Yoshibumi; Kobayashi, Keiko; Kobayashi, Toshihide; Sakurai, Jun; Terao, Yutaka; Nagahama, Masahiro

    2015-01-01

    Clostridium perfringens alpha-toxin elicits various immune responses such as the release of cytokines, chemokines, and superoxide via the GM1a/TrkA complex. Alpha-toxin possesses phospholipase C (PLC) hydrolytic activity that contributes to signal transduction in the pathogenesis of gas gangrene. Little is known about the relationship between lipid metabolism and TrkA activation by alpha-toxin. Using live-cell fluorescence microscopy, we monitored transbilayer movement of diacylglycerol (DAG) with the yellow fluorescent protein-tagged C1AB domain of protein kinase C-γ (EYFP-C1AB). DAG accumulated at the marginal region of the plasma membrane in alpha toxin-treated A549 cells, which also exhibited GM1a clustering and TrkA phosphorylation. Annexin V binding assays showed that alpha-toxin induced the exposure of phosphatidylserine on the outer leaflet of the plasma membrane. However, H148G, a variant toxin which binds cell membrane and has no enzymatic activity, did not induce DAG translocation, GM1a clustering, or TrkA phosphorylation. Alpha-toxin also specifically activated endogenous phospholipase Cγ-1 (PLCγ-1), a TrkA adaptor protein, via phosphorylation. U73122, an endogenous PLC inhibitor, and siRNA for PLCγ-1 inhibited the formation of DAG and release of IL-8. GM1a accumulation and TrkA phosphorylation in A549 cells treated with alpha-toxin were also inhibited by U73122. These results suggest that the flip-flop motion of hydrophobic lipids such as DAG leads to the accumulation of GM1a and TrkA. We conclude that the formation of DAG by alpha-toxin itself (first step) and activation of endogenous PLCγ-1 (second step) leads to alterations in membrane dynamics, followed by strong phosphorylation of TrkA. PMID:25910247

  6. Association of cord blood des-acyl ghrelin with birth weight, and placental GHS-R1 receptor expression in SGA, AGA, and LGA newborns.

    PubMed

    González-Domínguez, Martha I; Lazo-de-la-Vega-Monroy, Maria-Luisa; Zaina, Silvio; Sabanero, Myrna; Daza-Benítez, Leonel; Malacara, Juan Manuel; Barbosa-Sabanero, Gloria

    2016-07-01

    Although ghrelin in cord blood has been associated to birth weight, its role in fetal and postnatal growth has not been elucidated. The aim of this study was to analyze total ghrelin, acyl ghrelin (AG), and des-acyl ghrelin (DAG) in cord blood of newborns with idiopathic birth weight alterations, and to evaluate protein expression of placental GHS-R1, in order to investigate their correlation with birth weight and placental weight. We performed a cross-sectional comparative study in umbilical cord blood and placentas from healthy mothers of SGA, AGA, and LGA (small, adequate and large for gestational age) term newborns (n = 20 per group). Cord blood total ghrelin, AG, and DAG were measured by ELISA, and placental GHS-R1 expression was evaluated by Western blot. Cord blood DAG was higher in SGA compared to AGA newborns (902.1 ± 109.1 and 597.4 ± 58.2 pg/ml, respectively, p = 0.01) while LGA and AGA showed similar values (627.2 ± 76.4 pg/ml for LGA, p = 0.80). DAG negatively correlated with birthweight (r = -0.31, p = 0.02) and placental weight (r = -0.33, p = 0.02). No differences in AG or total ghrelin were found. GHS-R1 protein in placenta was not differentially expressed among SGA, AGA, and LGA. Our results suggest a role of DAG in intrauterine growth. Further studies are needed in order to elucidate the mechanisms by which DAG participates in fetal growth. PMID:26754660

  7. Membrane anchoring of diacylglycerol-lactones substituted with rigid hydrophobic acyl domains correlates with biological activities

    PubMed Central

    Raifman, Or; Kolusheva, Sofiya; Comin, Maria J.; Kedei, Noemi; Lewin, Nancy E.; Blumberg, Peter M.; Marquez, Victor E.; Jelinek, Raz

    2009-01-01

    Summary Synthetic diacylglycerol lactones (DAG-lactones) are effective modulators of critical cellular signaling pathways, downstream of the lipophilic second messenger diacylglycerol, that activate a host of protein kinase C (PKC) isozymes as well as other non-kinase proteins that share with PKC similar C1 membrane-targeting domains. A fundamental determinant of the biological activity of these amphiphilic molecules is the nature of their interactions with cellular membranes. This study characterizes the membrane interactions and bilayer anchoring of a series of DAG-lactones in which the hydrophobic moiety is a “molecular rod”, namely a rigid 4-[2-(R-phenyl)ethynyl]benzoate moiety in the acyl position. Application of assays employing chromatic biomimetic vesicles and biophysical techniques reveals that the mode of membrane anchoring of the DAG-lactone derivatives was markedly affected by the presence of the hydrophobic diphenyl rod and by the size of the functional unit displayed at the terminus of the rod. Two primary mechanisms of interaction were observed: surface binding of the DAG-lactones at the lipid/water interface and deep insertion of the ligands into the alkyl core of the lipid bilayer. These membrane-insertion properties could explain the different patterns of PKC translocation from cytosol to membranes induced by the molecular-rod DAG-lactones. This investigation emphasizes that the side-residues of DAG-lactones, rather than simply conferring hydrophobicity, profoundly influence membrane interactions and in that fashion may further contribute to the diversity of biological actions of these synthetic biomimetic ligands. PMID:19961537

  8. Dystroglycan Binding to α-Neurexin Competes with Neurexophilin-1 and Neuroligin in the Brain*

    PubMed Central

    Reissner, Carsten; Stahn, Johanna; Breuer, Dorothee; Klose, Martin; Pohlentz, Gottfried; Mormann, Michael; Missler, Markus

    2014-01-01

    α-Neurexins (α-Nrxn) are mostly presynaptic cell surface molecules essential for neurotransmission that are linked to neuro-developmental disorders as autism or schizophrenia. Several interaction partners of α-Nrxn are identified that depend on alternative splicing, including neuroligins (Nlgn) and dystroglycan (αDAG). The trans-synaptic complex with Nlgn1 was extensively characterized and shown to partially mediate α-Nrxn function. However, the interactions of α-Nrxn with αDAG, neurexophilins (Nxph1) and Nlgn2, ligands that occur specifically at inhibitory synapses, are incompletely understood. Using site-directed mutagenesis, we demonstrate the exact binding epitopes of αDAG and Nxph1 on Nrxn1α and show that their binding is mutually exclusive. Identification of an unusual cysteine bridge pattern and complex type glycans in Nxph1 ensure binding to the second laminin/neurexin/sex hormone binding (LNS2) domain of Nrxn1α, but this association does not interfere with Nlgn binding at LNS6. αDAG, in contrast, interacts with both LNS2 and LNS6 domains without inserts in splice sites SS#2 or SS#4 mostly via LARGE (like-acetylglucosaminyltransferase)-dependent glycans attached to the mucin region. Unexpectedly, binding of αDAG at LNS2 prevents interaction of Nlgn at LNS6 with or without splice insert in SS#4, presumably by sterically hindering each other in the u-form conformation of α-Nrxn. Thus, expression of αDAG and Nxph1 together with alternative splicing in Nrxn1α may prevent or facilitate formation of distinct trans-synaptic Nrxn·Nlgn complexes, revealing an unanticipated way to contribute to the identity of synaptic subpopulations. PMID:25157101

  9. Prefission Constriction of Golgi Tubular Carriers Driven by Local Lipid Metabolism: A Theoretical Model

    PubMed Central

    Shemesh, Tom; Luini, Alberto; Malhotra, Vivek; Burger, Koert N. J.; Kozlov, Michael M.

    2003-01-01

    Membrane transport within mammalian cells is mediated by small vesicular as well as large pleiomorphic transport carriers (TCs). A major step in the formation of TCs is the creation and subsequent narrowing of a membrane neck connecting the emerging carrier with the initial membrane. In the case of small vesicular TCs, neck formation may be directly induced by the coat proteins that cover the emerging vesicle. However, the mechanism underlying the creation and narrowing of a membrane neck in the generation of large TCs remains unknown. We present a theoretical model for neck formation based on the elastic model of membranes. Our calculations suggest a lipid-driven mechanism with a central role for diacylglycerol (DAG). The model is applied to a well-characterized in vitro system that reconstitutes TC formation from the Golgi complex, namely the pearling and fission of Golgi tubules induced by CtBP/BARS, a protein that catalyzes the conversion of lysophosphatidic acid into phosphatidic acid. In view of the importance of a PA-DAG cycle in the formation of Golgi TCs, we assume that the newly formed phosphatidic acid undergoes rapid dephosphorylation into DAG. DAG possesses a unique molecular shape characterized by an extremely large negative spontaneous curvature, and it redistributes rapidly between the membrane monolayers and along the membrane surface. Coupling between local membrane curvature and local lipid composition results, by mutual enhancement, in constrictions of the tubule into membrane necks, and a related inhomogeneous lateral partitioning of DAG. Our theoretical model predicts the exact dimensions of the constrictions observed in the pearling Golgi tubules. Moreover, the model is able to explain membrane neck formation by physiologically relevant mole fractions of DAG. PMID:14645071

  10. Involvement of diacylglycerol produced by phospholipase D activation in Aβ-induced reduction of sAPPα secretion in SH-SY5Y neuroblastoma cells.

    PubMed

    Tanabe, Fuminori; Nakajima, Tomoko; Ito, Masahiko

    2014-04-18

    We previously reported that the thiol proteinase inhibitor, E-64-d, ameliorated amyloid β (Aβ)-induced reduction of soluble amyloid precursor protein α (sAPPα) secretion by reversing ceramide-induced protein kinase C down-regulation in SH-SY5Y neuroblastoma cells. In the present study, we showed that Aβ (1-42) peptide enhanced diacylglycerol (DAG) production by phospholipase D (PLD) activation in these cells. We subsequently examined whether PLD was involved in Aβ-induced reduction of sAPPα secretion and showed that 2 μM CAY10593, which selectively inhibits PLD2, ameliorated reduction of sAPPα secretion, whereas 50 nM CAY10593, which selectively inhibits PLD1, did not. Moreover, 50 µM propranolol, a phosphatidic acid phosphohydrolase inhibitor, also ameliorated Aβ-induced reduction of sAPPα secretion, suggesting that DAG may be responsible for Aβ-induced reduction of sAPPα. We subsequently examined whether DAG affects sAPPα secretion and showed that a DAG analog reduced sAPPα secretion in SH-SY5Y cells. In addition, DAG enhanced ceramide production by stimulating neutral sphingomyelinase (N-SMase) activity. We previously demonstrated that Aβ stimulates N-SMase activity in SH-SY5Y cells. Here, we showed that inhibition of PLD2 by 2 μM CAY10593 suppressed Aβ-induced N-SMase activation. Taken together, the results suggest that DAG produced through the PLD pathway is involved in Aβ-induced reduction of sAPPα secretion in SH-SY5Y cells. PMID:24650665

  11. The Updated Bottom Up Solution applied to mass spectrometry of soybean oil in a dietary supplement gelcap.

    PubMed

    Byrdwell, William Craig

    2015-07-01

    Among the goals of lipidomics applied to triacylglycerols (TAGs) is identification of molecular species, degree and location of unsaturation, and positions of fatty acyl chains (i.e., identification of regioisomers). Toward those ends, we define one, two, and three "Critical Ratios" for Types I, II, and III TAGs that provided different aspects of the desired information. Critical Ratio 1, [MH](+)/Σ[DAG](+), is correlated to the degree of unsaturation ([MH](+) is the protonated molecule and Σ[DAG](+) is the sum of diacylglycerol-like ions, [DAG](+)); Critical Ratio 2, [AA](+)/[AB](+) for Type II TAGs ("ABA/AAB/BAA") and [AC](+)/([AB](+)+[BC](+)) for Type III TAGs ("ABC/CBA/BAC/CAB/ACB/BCA"), is correlated to identification of regioisomers; and Critical Ratio 3, [BC](+)/[AB](+), provides information about those [DAG](+) from Type III TAGs. Furthermore, Critical Ratios are used in the Updated Bottom Up Solution (UBUS) to reproduce the mass spectra of TAGs by atmospheric pressure chemical ionization mass spectrometry applied to analysis of soybean oil in a dietary supplement gelcap. We present a new model for the [MH](+)/Σ[DAG](+) ratio, quantify regioisomers using the [AA](+)/[AB](+) ratio, and describe trends for [BC](+)/[AB](+) that have never been reported before. The UBUS is also applied to other classes of molecules, i.e., vitamin D and DAGs. The amount of vitamin D3 in the gelcap fell from 2011 ± 22 when received to 1689 ± 33 just prior to expiration. The Critical Ratios constitute a compact data set that can provide structural information and also act as a library of mass spectra. PMID:25855147

  12. A dual-purpose silver nanoparticles biosynthesized using aqueous leaf extract of Detarium microcarpum: An under-utilized species.

    PubMed

    Labulo, Ayomide H; Adesuji, Elijah T; Dedeke, Oyinade A; Bodede, Olusola S; Oseghale, Charles O; Moodley, Roshila; Nyamori, Vincent O; Dare, Enock O; Adegoke, Olajire A

    2016-11-01

    The need for green synthesis of emerging industrial materials has led to the biosynthesis of nanoparticles from plants to circumvent the adverse by-products of chemical synthesis. In this study, the leaf extract of Detarium mirocarpum Guill & Perr, a small tree belonging to the family Fabaceae (Legume), was used to synthesize silver nanoparticles (DAgNPs). DAgNPs were characterized using spectroscopic techniques (Ultraviolet-Visible spectroscopy and Fourier Transform Infrared spectroscopy) which showed hydroxyl and carbonyl functional groups to be responsible for their synthesis. DAgNPs were observed to be crystalline and spherical. The average size, determined by transmission electron microscopy (TEM) was 17.05nm. The antioxidant activity of DAgNPs ranked from moderate to good. The ability of DAgNPs to sense Hg(2+) and Fe(3+) ions in aqueous medium was also investigated. The quenching of the SPR peak at 430nm was used to monitor the toxic and heavy metal ions with linear ranges of 20-70µgmL(-1) and 10-40µgmL(-1) for Hg(2+) and Fe(3+), respectively. The limit of detection (LOD) and limit of quantification (LOQ) obtained for Hg(2+) was 2.05µgmL(-1) and 6.21µgmL(-1), respectively and for Fe(3+) was 5.01µgmL(-1) and 15.21µgmL(-1), respectively. The intra- and inter-day assessments of accuracy and repeatability gave relative errors less than 1% in all instances. DAgNPs can therefore provide a convenient method of sensing the toxic metals easily. PMID:27591670

  13. Altered Skeletal Muscle Lipase Expression and Activity Contribute to Insulin Resistance in Humans

    PubMed Central

    Badin, Pierre-Marie; Louche, Katie; Mairal, Aline; Liebisch, Gerhard; Schmitz, Gerd; Rustan, Arild C.; Smith, Steven R.; Langin, Dominique; Moro, Cedric

    2011-01-01

    OBJECTIVE Insulin resistance is associated with elevated content of skeletal muscle lipids, including triacylglycerols (TAGs) and diacylglycerols (DAGs). DAGs are by-products of lipolysis consecutive to TAG hydrolysis by adipose triglyceride lipase (ATGL) and are subsequently hydrolyzed by hormone-sensitive lipase (HSL). We hypothesized that an imbalance of ATGL relative to HSL (expression or activity) may contribute to DAG accumulation and insulin resistance. RESEARCH DESIGN AND METHODS We first measured lipase expression in vastus lateralis biopsies of young lean (n = 9), young obese (n = 9), and obese-matched type 2 diabetic (n = 8) subjects. We next investigated in vitro in human primary myotubes the impact of altered lipase expression/activity on lipid content and insulin signaling. RESULTS Muscle ATGL protein was negatively associated with whole-body insulin sensitivity in our population (r = −0.55, P = 0.005), whereas muscle HSL protein was reduced in obese subjects. We next showed that adenovirus-mediated ATGL overexpression in human primary myotubes induced DAG and ceramide accumulation. ATGL overexpression reduced insulin-stimulated glycogen synthesis (−30%, P < 0.05) and disrupted insulin signaling at Ser1101 of the insulin receptor substrate-1 and downstream Akt activation at Ser473. These defects were fully rescued by nonselective protein kinase C inhibition or concomitant HSL overexpression to restore a proper lipolytic balance. We show that selective HSL inhibition induces DAG accumulation and insulin resistance. CONCLUSIONS Altogether, the data indicate that altered ATGL and HSL expression in skeletal muscle could promote DAG accumulation and disrupt insulin signaling and action. Targeting skeletal muscle lipases may constitute an interesting strategy to improve insulin sensitivity in obesity and type 2 diabetes. PMID:21498783

  14. Progesterone induces meiotic division in the amphibian oocyte by releasing lipid second messengers from the plasma membrane.

    PubMed

    Morrill, G A; Kostellow, A B

    1999-01-01

    Meiosis in the amphibian oocyte is normally initiated by gonadotropins, which stimulate follicle cells to secret progesterone. The progesterone-induced G2/M transition in the amphibian oocyte was the first well-defined example of a steroid effect at the plasma membrane, since it could be shown that exogenous, but not injected, progesterone induced meiosis and that many of the progesterone-induced changes associated with meiosis occurred in enucleated oocytes. We find that [3H]progesterone binding to isolated plasma membranes of Rana pipiens oocytes is saturable, specific and temperature-dependent. Photoaffinity labeling with the synthetic progestin [3H]R5020 followed by gel electrophoresis demonstrated progestin binding to both 80 and 110 kDa proteins in the oocyte cytosol, whereas only the 110 kDa R5020 binding protein was present in the oocyte plasma membrane. We have shown that progesterone acts at Rana oocyte plasma membrane receptors within seconds to release a cascade of lipid messengers. Membrane-receptor binding causes the successive activation of: 1) N-methyltransferases, which convert phosphatidylethanolamine to phosphatidylcholine (PC); 2) an exchange reaction between PC and ceramide to form sphingomyelin (SM) and 1,2-diacylglycerol (DAG); 3) phospholipase D/phosphatidate phosphohydrolase, releasing a second DAG transient; and 4) phosphatidylinositol-specific phospholipase C, generating inositol trisphosphate and a third DAG transient. Within minutes, diglyceride kinase converts newly formed DAG species to phosphatidic acid, turning off the successive DAG signals. A transient fall (0-30 s) in intracellular ceramide is followed (within 1-2 min) by a sustained rise in intracellular ceramide lasting 3-4 h. This ceramide may be significant in later cyclin-dependent steps. We conclude that the initial action of progesterone at its plasma membrane receptor triggers a series of enzyme activations that modify the membrane and release multiple DAG species. PMID

  15. Clostridium perfringens Alpha-Toxin Induces Gm1a Clustering and Trka Phosphorylation in the Host Cell Membrane

    PubMed Central

    Takagishi, Teruhisa; Oda, Masataka; Kabura, Michiko; Kurosawa, Mie; Tominaga, Kaori; Urano, Shiori; Ueda, Yoshibumi; Kobayashi, Keiko; Kobayashi, Toshihide; Sakurai, Jun; Terao, Yutaka; Nagahama, Masahiro

    2015-01-01

    Clostridium perfringens alpha-toxin elicits various immune responses such as the release of cytokines, chemokines, and superoxide via the GM1a/TrkA complex. Alpha-toxin possesses phospholipase C (PLC) hydrolytic activity that contributes to signal transduction in the pathogenesis of gas gangrene. Little is known about the relationship between lipid metabolism and TrkA activation by alpha-toxin. Using live-cell fluorescence microscopy, we monitored transbilayer movement of diacylglycerol (DAG) with the yellow fluorescent protein-tagged C1AB domain of protein kinase C-γ (EYFP-C1AB). DAG accumulated at the marginal region of the plasma membrane in alpha toxin-treated A549 cells, which also exhibited GM1a clustering and TrkA phosphorylation. Annexin V binding assays showed that alpha-toxin induced the exposure of phosphatidylserine on the outer leaflet of the plasma membrane. However, H148G, a variant toxin which binds cell membrane and has no enzymatic activity, did not induce DAG translocation, GM1a clustering, or TrkA phosphorylation. Alpha-toxin also specifically activated endogenous phospholipase Cγ-1 (PLCγ-1), a TrkA adaptor protein, via phosphorylation. U73122, an endogenous PLC inhibitor, and siRNA for PLCγ-1 inhibited the formation of DAG and release of IL-8. GM1a accumulation and TrkA phosphorylation in A549 cells treated with alpha-toxin were also inhibited by U73122. These results suggest that the flip-flop motion of hydrophobic lipids such as DAG leads to the accumulation of GM1a and TrkA. We conclude that the formation of DAG by alpha-toxin itself (first step) and activation of endogenous PLCγ-1 (second step) leads to alterations in membrane dynamics, followed by strong phosphorylation of TrkA. PMID:25910247

  16. Simultaneous detection of Ca2+ and diacylglycerol signaling in living cells.

    PubMed

    Tewson, Paul; Westenberg, Mara; Zhao, Yongxin; Campbell, Robert E; Quinn, Anne Marie; Hughes, Thomas E

    2012-01-01

    Phospholipase C produces two second messengers--diacylglycerol (DAG), which remains in the membrane, and inositol triphosphate (IP(3)), which triggers the release of calcium ions (Ca(2+)) from intracellular stores. Genetically encoded sensors based on a single circularly permuted fluorescent protein (FP) are robust tools for studying intracellular Ca(2+) dynamics. We have developed a robust sensor for DAG based on a circularly permuted green FP that can be co-imaged with the red fluorescent Ca(2+) sensor R-GECO for simultaneous measurement of both second messengers. PMID:22912738

  17. Spectral and electrochemical detection of protonated triplex formation by a small-molecule anticancer agent

    NASA Astrophysics Data System (ADS)

    Feng, Lingyan; Li, Xi; Peng, Yinghua; Geng, Jie; Ren, Jinsong; Qu, Xiaogang

    2009-10-01

    Triplex helical formation has been the focus of considerable interest because of possible applications in developing new molecular biology tools as well as therapeutic agents and the possible relevance of H-DNA structures in biology system. We report here that a small-molecule anticancer agent, coralyne, has binding preference to the less stable protonated triplex d(C +-T) 6:d(A-G) 6·d(C-T) 6 over duplex d(A-G) 6·d(C-T) 6 and shows different spectral and electrochemical characteristics when binding to triplex and duplex DNA, indicating that electrochemical technique can detect the less stable protonated triplex formation.

  18. Lipidomic and transcriptomic analyses of Chlamydomonas reinhardtii under heat stress unveil a direct route for the conversion of membrane lipids into storage lipids.

    PubMed

    Légeret, B; Schulz-Raffelt, M; Nguyen, H M; Auroy, P; Beisson, F; Peltier, G; Blanc, G; Li-Beisson, Y

    2016-04-01

    Studying how photosynthetic cells modify membrane lipids in response to heat stress is important to understand how plants and microalgae adapt to daily fluctuations in temperature and to investigate new lipid pathways. Here, we investigate changes occurring in lipid molecular species and lipid metabolism genes during early response to heat stress in the model photosynthetic microorganism Chlamydomonas reinhardtii. Lipid molecular species analyses revealed that, after 60 min at 42 °C, a strong decrease in specific polyunsaturated membrane lipids was observed together with an increase in polyunsaturated triacylglycerols (TAGs) and diacylglycerols (DAGs). The fact that decrease in the major chloroplastic monogalactosyldiacylglycerol sn1-18:3/sn2-16:4 was mirrored by an accumulation of DAG sn1-18:3/sn2-16:4 and TAG sn1-18:3/sn2-16:4/sn3-18:3 indicated that newly accumulated TAGs were formed via direct conversion of monogalactosyldiacylglycerols to DAGs then TAGs. Lipidomic analyses showed that the third fatty acid of a TAG likely originated from a phosphatidylethanolamine or a diacylglyceryl-O-4'-(N,N,N,-trimethyl)-homoserine betaine lipid species. Candidate genes for this TAG synthesis pathway were provided through comparative transcriptomic analysis and included a phospholipase A2 homolog and the DAG acyltransferase DGTT1. This study gives insights into the molecular events underlying changes in membrane lipids during heat stress and reveals an alternative route for TAG synthesis. PMID:26477535

  19. 78 FR 56737 - Privacy Act of 1974; System of Records

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-09-13

    ... Force Evaluation and Reporting System,'' JUSTICE/DAG-003, last published in its entirety at 57 FR 8473... Forces (OCDETF), Department of Justice. ACTION: Notice of a modified system of records. SUMMARY: Pursuant... Task Forces Management Information System'' (OCDETF MIS), changing the system number from...

  20. Diacylglycerol kinases in membrane trafficking

    PubMed Central

    Xie, Shuwei; Naslavsky, Naava; Caplan, Steve

    2015-01-01

    Diacylglycerol kinases (DGKs) belong to a family of cytosolic kinases that regulate the phosphorylation of diacylglycerol (DAG), converting it into phosphatidic acid (PA). There are 10 known mammalian DGK isoforms, each with a different tissue distribution and substrate specificity. These differences allow regulation of cellular responses by fine-tuning the delicate balance of cellular DAG and PA. DGK isoforms are best characterized as mediators of signal transduction and immune function. However, since recent studies reveal that DAG and PA are also involved in the regulation of endocytic trafficking, it is therefore anticipated that DGKs also plays an important role in membrane trafficking. In this review, we summarize the literature discussing the role of DGK isoforms at different stages of endocytic trafficking, including endocytosis, exocytosis, endocytic recycling, and transport from/to the Golgi apparatus. Overall, these studies contribute to our understanding of the involvement of PA and DAG in endocytic trafficking, an area of research that is drawing increasing attention in recent years. PMID:27057419

  1. Operation and Maintenance of the 35mm Filmstrip Projector.

    ERIC Educational Resources Information Center

    Rampino, Tony

    A manual format is used to explain: (1) the parts of a filmstrip projector, (2) threading procedures, (3) minor maintenance and repair techniques, and (4) purchasing hints. One-half of the manual is devoted to illustrations of equipment and material. (DAG)

  2. Highly efficient solvent-free synthesis of 1,3-diacylglycerols by lipase immobilised on nano-sized magnetite particles.

    PubMed

    Meng, Xiao; Xu, Gang; Zhou, Qin-Li; Wu, Jian-Ping; Yang, Li-Rong

    2014-01-15

    Recently, 1,3-DAGs (1,3-diacylglycerols) have attracted considerable attention as healthy components of food, oil and pharmaceutical intermediates. Generally, 1,3-DAG is prepared by lipase-mediated catalysis in a solvent free system. However, the system's high reaction temperature (required to reach the reactants' melting point), high substrate concentration and high viscosity severely reduce the lipase's activity, selectivity and recycling efficiency. In this report, MjL (Mucor javanicus lipase) was found to have the best performance in the solvent-free synthesis of 1,3-DAGs of several common commercial lipases. By covalent binding to amino-group-activated NSM (nano-sized magnetite) particles and cross-linking to form an enzyme aggregate coat, MjL's specific activity increased 10-fold, and was able to be reused for 10 cycles with 90% residual activity at 55°C. 1,3-DAGs of lauric, myristic, palmitic, stearic, oleic and linoleic acid were prepared using the resulting immobilised enzyme, all with yields greater than 90%, and the reaction time was also greatly reduced. PMID:24054246

  3. Diacylglycerol levels modulate the cellular distribution of the nicotinic acetylcholine receptor.

    PubMed

    Kamerbeek, Constanza B; Mateos, Melina V; Vallés, Ana S; Pediconi, María F; Barrantes, Francisco J; Borroni, Virginia

    2016-05-01

    Diacylglycerol (DAG), a second messenger involved in different cell signaling cascades, activates protein kinase C (PKC) and D (PKD), among other kinases. The present work analyzes the effects resulting from the alteration of DAG levels on neuronal and muscle nicotinic acetylcholine receptor (AChR) distribution. We employ CHO-K1/A5 cells, expressing adult muscle-type AChR in a stable manner, and hippocampal neurons, which endogenously express various subtypes of neuronal AChR. CHO-K1/A5 cells treated with dioctanoylglycerol (DOG) for different periods showed augmented AChR cell surface levels at short incubation times (30min-4h) whereas at longer times (18h) the AChR was shifted to intracellular compartments. Similarly, in cultured hippocampal neurons surface AChR levels increased as a result of DOG incubation for 4h. Inhibition of endogenous DAG catabolism produced changes in AChR distribution similar to those induced by DOG treatment. Specific enzyme inhibitors and Western blot assays revealed that DAGs exert their effect on AChR distribution through the modulation of the activity of classical PKC (cPKC), novel PKC (nPKC) and PKD activity. PMID:26898898

  4. Identification of a soluble phosphatidate phosphohydrolase in the developing cotyledons of Morordica charantia

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phosphatidate phosphatase (3-sn-phosphatidate phosphohydrolase, EC 3.1.3.4), which is also known as PAP, catalyzes the dephosphorylation of phosphatidate (PtdOH) to form diacylglycerol (DAG) and inorganic phosphate. In eukaryotes, PAP driven reaction is the committed step in the synthesis of triacy...

  5. Book Review: Space Weather: Physics and Effects

    NASA Astrophysics Data System (ADS)

    Wilkinson, Phil

    2007-11-01

    At 438 pages, Space Weather: Physics and Effects, edited by Volker Bothmer and Ioannis A. Daglis, seems like a daunting read. But its thickness belies its conversational tone, and its content provides a different presentation of material aimed at drawing in a new audience while satisfying the present space weather audience's interest in their subject. I found reading this book a pleasure.

  6. Mass spectrometry of the lithium adducts of diacylglycerols containing hydroxy FA in castor oil and two normal FA

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Castor oil can be used in industry. The molecular species of triacylglycerols containing hydroxy fatty acids (FA) in castor oil have been identified. We report here the identification of twelve diacylglycerols (DAG) containing hydroxy FA in castor oil using positive ion electrospray ionization mass ...

  7. Quantification of the molecular species of diacylglycerols,triacylglycerols and tetraacylglycerols in lesquerella (Physaria fendleri) oil by HPLC and MS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Ten diacylglycerols (DAG), 74 triacylglycerols (TAG) and 13 tetraacylglycerols in the seed oil of Physaria fendleri were recently identified by HPLC and MS. These acylglycerols (AG) were quantified by HPLC with evaporative light scattering detector and electrospray ionization mass spectrometry of th...

  8. Synthesis of monoacylglycerol containing pinolenic acid via stepwise esterification using a cold active lipase.

    PubMed

    Pyo, Young-Gil; Hong, Seung In; Kim, Yangha; Kim, Byung Hee; Kim, In-Hwan

    2012-01-01

    High purity monoacylglycerol (MAG) containing pinolenic acid was synthesized via stepwise esterification of glycerol and fatty acids from pine nut oil using a cold active lipase from Penicillium camembertii as a biocatalyst. Effects of temperature, molar ratio, water content, enzyme loading, and vacuum on the synthesis of MAG by lipase-catalyzed esterification of glycerol and fatty acid from pine nut oil were investigated. Diacylglycerol (DAG) as well as MAG increased significantly when temperature was increased from 20 to 40 °C. At a molar ratio of 1:1, MAG content decreased because of the significant increase in DAG content. Water has a profound influence on both MAG and DAG content through the entire course of reaction. The reaction rate increased significantly as enzyme loading increased up to 600 units. Vacuum was an effective method to reduce DAG content. The optimum temperature, molar ratio, water content, enzyme loading, vacuum, and reaction time were 20 °C, 1:5 (fatty acid to glycerol), 2%, 600 units, 5 torr, and 24 h, respectively. MAG content further increased via lipase-catalyzed second step esterification at subzero temperature. P. camembertii lipase exhibited esterification activity up to -30 °C. PMID:22753389

  9. Unleashing mesenchymal chemotaxis.

    PubMed

    Aguilar-Cuenca, Rocío; Vicente-Manzanares, Miguel

    2014-12-22

    In this issue of Developmental Cell, Asokan and colleagues (2014) report that the phospholipase Cγ (PLCγ)-diacyl glycerol (DAG) protein kinase Cα (PKCα) signaling axis inhibits actomyosin bundling. This preferentially occurs at the leading edge of chemotactic mesenchymal cells via noncanonical phosphorylation of the regulatory light chain (RLC) of nonmuscle myosin II. PMID:25535912

  10. Algorithmic height compression of unordered trees.

    PubMed

    Ben-Naoum, Farah; Godin, Christophe

    2016-01-21

    By nature, tree structures frequently present similarities between their sub-parts. Making use of this redundancy, different types of tree compression techniques have been designed in the literature to reduce the complexity of tree structures. A popular and efficient way to compress a tree consists of merging its isomorphic subtrees, which produces a directed acyclic graph (DAG) equivalent to the original tree. An important property of this method is that the compressed structure (i.e. the DAG) has the same height as the original tree, thus limiting partially the possibility of compression. In this paper we address the problem of further compressing this DAG in height. The difficulty is that compression must be carried out on substructures that are not exactly isomorphic as they are strictly nested within each-other. We thus introduced a notion of quasi-isomorphism between subtrees that makes it possible to define similar patterns along any given path in a tree. We then proposed an algorithm to detect these patterns and to merge them, thus leading to compressed structures corresponding to DAGs augmented with return edges. In this way, redundant information is removed from the original tree in both width and height, thus achieving minimal structural compression. The complete compression algorithm is then illustrated on the compression of various plant-like structures. PMID:26551155

  11. Structure and mechanism of an intramembrane liponucleotide synthetase central for phospholipid biosynthesis

    PubMed Central

    Liu, Xiuying; Yin, Yan; Wu, Jinjun; Liu, Zhenfeng

    2014-01-01

    Phospholipids are elemental building-block molecules for biological membranes. Biosynthesis of phosphatidylinositol, phosphatidylglycerol and phosphatidylserine requires a central liponucleotide intermediate named cytidine-diphosphate diacylglycerol (CDP-DAG). The CDP-DAG synthetase (Cds) is an integral membrane enzyme catalysing the formation of CDP-DAG, an essential step for phosphoinositide recycling during signal transduction. Here we report the structure of the Cds from Thermotoga maritima (TmCdsA) at 3.4 Å resolution. TmCdsA forms a homodimer and each monomer contains nine transmembrane helices arranged into a novel fold with three domains. An unusual funnel-shaped cavity penetrates half way into the membrane, allowing the enzyme to simultaneously accept hydrophilic substrate (cytidine 5′-triphosphate (CTP)/deoxy-CTP) from cytoplasm and hydrophobic substrate (phosphatidic acid) from membrane. Located at the bottom of the cavity, a Mg2+-K+ hetero-di-metal centre coordinated by an Asp-Asp dyad serves as the cofactor of TmCdsA. The results suggest a two-metal-ion catalytic mechanism for the Cds-mediated synthesis of CDP-DAG at the membrane–cytoplasm interface. PMID:24968740

  12. Abrogating monoacylglycerol acyltransferase activity in liver improves glucose tolerance and hepatic insulin signaling in obese mice.

    PubMed

    Hall, Angela M; Soufi, Nisreen; Chambers, Kari T; Chen, Zhouji; Schweitzer, George G; McCommis, Kyle S; Erion, Derek M; Graham, Mark J; Su, Xiong; Finck, Brian N

    2014-07-01

    Monoacylglycerol acyltransferase (MGAT) enzymes convert monoacylglycerol to diacylglycerol (DAG), a lipid that has been linked to the development of hepatic insulin resistance through activation of protein kinase C (PKC). The expression of genes that encode MGAT enzymes is induced in the livers of insulin-resistant human subjects with nonalcoholic fatty liver disease, but whether MGAT activation is causal of hepatic steatosis or insulin resistance is unknown. We show that the expression of Mogat1, which encodes MGAT1, and MGAT activity are also increased in diet-induced obese (DIO) and ob/obmice. To probe the metabolic effects of MGAT1 in the livers of obese mice, we administered antisense oligonucleotides (ASOs) against Mogat1 to DIO and ob/ob mice for 3 weeks. Knockdown of Mogat1 in liver, which reduced hepatic MGAT activity, did not affect hepatic triacylglycerol content and unexpectedly increased total DAG content. Mogat1 inhibition also increased both membrane and cytosolic compartment DAG levels. However, Mogat1 ASO treatment significantly improved glucose tolerance and hepatic insulin signaling in obese mice. In summary, inactivation of hepatic MGAT activity, which is markedly increased in obese mice, improved glucose tolerance and hepatic insulin signaling independent of changes in body weight, intrahepatic DAG and TAG content, and PKC signaling. PMID:24595352

  13. Regulation of cellular diacylglycerol through lipid phosphate phosphatases is required for pathogenesis of the rice blast fungus, Magnaporthe oryzae.

    PubMed

    Sadat, Md Abu; Jeon, Junhyun; Mir, Albely Afifa; Choi, Jaeyoung; Choi, Jaehyuk; Lee, Yong-Hwan

    2014-01-01

    Considering implication of diacylglycerol in both metabolism and signaling pathways, maintaining proper levels of diacylglycerol (DAG) is critical to cellular homeostasis and development. Except the PIP2-PLC mediated pathway, metabolic pathways leading to generation of DAG converge on dephosphorylation of phosphatidic acid catalyzed by lipid phosphate phosphatases. Here we report the role of such enzymes in a model plant pathogenic fungus, Magnaporthe oryzae. We identified five genes encoding putative lipid phosphate phosphatases (MoLPP1 to MoLPP5). Targeted disruption of four genes (except MoLPP4) showed that MoLPP3 and MoLPP5 are required for normal progression of infection-specific development and proliferation within host plants, whereas MoLPP1 and MoLPP2 are indispensable for fungal pathogenicity. Reintroduction of MoLPP3 and MoLPP5 into individual deletion mutants restored all the defects. Furthermore, exogenous addition of saturated DAG not only restored defect in appressorium formation but also complemented reduced virulence in both mutants. Taken together, our data indicate differential roles of lipid phosphate phosphatase genes and requirement of proper regulation of cellular DAGs for fungal development and pathogenesis. PMID:24959955

  14. Exercise-induced cardiac performance in autoimmune (type 1) diabetes is associated with a decrease in myocardial diacylglycerol.

    PubMed

    Loganathan, Rajprasad; Novikova, Lesya; Boulatnikov, Igor G; Smirnova, Irina V

    2012-09-01

    One of the fundamental biochemical defects underlying the complications of diabetic cardiovascular system is elevation of diacylglycerol (DAG) and its effects on protein kinase C (PKC) signaling. It has been noted that exercise training attenuates poor cardiac performance in Type 1 diabetes. However, the role of PKC signaling in exercise-induced alleviation of cardiac abnormalities in diabetes is not clear. We investigated the possibility that exercise training modulates PKC-βII signaling to elicit its beneficial effects on the diabetic heart. bio-breeding diabetic resistant rats, a model reminiscent of Type 1 diabetes in humans, were randomly assigned to four groups: 1) nonexercised nondiabetic (NN); 2) nonexercised diabetic (ND); 3) exercised nondiabetic; and 4) exercised diabetic. Treadmill training was initiated upon the onset of diabetes. At the end of 8 wk, left ventricular (LV) hemodynamic assessment revealed compromised function in ND compared with the NN group. LV myocardial histology revealed increased collagen deposition in ND compared with the NN group, while electron microscopy showed a reduction in the viable mitochondrial fraction. Although the PKC-βII levels and activity were unchanged in the diabetic heart, the DAG levels were increased. With exercise training, the deterioration of LV structure and function in diabetes was attenuated. Notably, improved cardiac performance in training was associated with a decrease in myocardial DAG levels in diabetes. Exercise-induced benefits on cardiac performance in diabetes may be mediated by prevention of an increase in myocardial DAG levels. PMID:22797313

  15. Relative oxidative stability of diacylglycerol and triacylglycerol oils.

    PubMed

    Qi, Jin F; Wang, Xiang Y; Shin, Jung-Ah; Lee, Young-Hwa; Jang, Young-Seok; Lee, Jeung Hee; Hong, Soon-Taek; Lee, Ki-Teak

    2015-03-01

    To compare the oxidative stability between diacylglycerol (DAG) oil and conventional triacylglycerol (TAG) oil (that is, soybean oil), the prepared stripped diacylglycerol oil (SDO) and soybean oil (SSBO) were stored at 60 °C in the dark for 144 h. During storage peroxide values (POVs), contents of aldehydes, unsaturated fatty acids were measured to evaluate the oxidative stabilities of the 2 oils. The results showed the content of C18:2, C18:3, and total unsaturated fatty acid decreased faster in DAG oil than in soybean oil, whereas the decreased rate of C18:1 was similar in 2 oils. Also, both rate constants (K1 and K2) obtained from POV (K1 ) and total aldehydes (K2 ) indicated that DAG oil (K1 = 3.22 mmol/mol FA h(-1) , K2 = 0.023 h(-1)) was oxidized more rapidly than soybean oil (K1 = 2.56 mmol/mol FA h(-1) , K2 = 0.021 h(-1)), which was mainly due to the difference of acylglycerol composition of the 2 oils along with higher C18:3 (9.6%) in SDO than SSBO (5.7%). It is concluded that DAG was more easily oxidized than soybean oil at 60 °C in the dark for 144 h. PMID:25678328

  16. PKC-2 phosphorylation of UNC-18 Ser322 in AFD neurons regulates temperature dependency of locomotion.

    PubMed

    Edwards, Mark R; Johnson, James R; Rankin, Kimberley; Jenkins, Rosalind E; Maguire, Carl; Morgan, Alan; Burgoyne, Robert D; Barclay, Jeff W

    2012-05-16

    Diacylglycerol (DAG)/protein kinase C (PKC) signaling plays an integral role in the regulation of neuronal function. This is certainly true in Caenorhabditis elegans and in particular for thermosensory signaling and behavior. Downstream molecular targets for transduction of this signaling cascade remain, however, virtually uncharacterized. We investigated whether PKC phosphorylation of Munc18-1, an essential protein in vesicle trafficking and exocytosis, was the downstream effector for DAG regulation of thermosensory behavior. We demonstrate here that the C. elegans ortholog of Munc18-1, UNC-18, was phosphorylated in vitro at Ser322. Transgenic rescue of unc-18-null worms with Ser322 phosphomutants displayed altered thermosensitivity. C. elegans expresses three DAG-regulated PKCs, and blocking UNC-18 Ser322 phosphorylation was phenocopied only by deletion of calcium-activated PKC-2. Expression of nonphosphorylatable UNC-18 S322A, either pan-neuronally or specifically in AFD thermosensory neurons, converted wild-type worms to a pkc-2-null phenotype. These data demonstrate that an individual DAG-dependent thermosensory behavior of an organism is effected specifically by the downstream PKC-2 phosphorylation of UNC-18 on Ser322 in AFD neurons. PMID:22593072

  17. Dominant Achievement Goals across Tracks in High School

    ERIC Educational Resources Information Center

    Scheltinga, Peter A. M.; Kuyper, Hans; Timmermans, Anneke C.; van der Werf, Greetje P. C.

    2016-01-01

    The dominant achievement goals (DAGs) of 7,008 students in the third grade of Dutch secondary education (US grade 9) were investigated, based on Elliot & McGregors' 2 × 2 framework (2001), in relation to track-level and motivational variables. We found the mastery-approach goal and the performance-approach goal, generally considered adaptive,…

  18. Structural analysis of galactoarabinan from duckweed.

    PubMed

    Yu, Li; Yu, Changjiang; Zhu, Ming; Cao, Yingping; Yang, Haiyan; Zhang, Xu; Ma, Yubin; Zhou, Gongke

    2015-03-01

    A highly branched galactoarabinan named DAG1 (Mw∼4.0×10(4) Da) was purified from Lemna aequinoctialis 6000 via 70% (v/v) ethanol extraction, followed by size-exclusion chromatography on Bio-Gel P2 and Superdex 75. Methylation analysis showed that DAG1 consisted of t-Araf, (1→5)-Araf, (1→2,5)-Araf, (1→3)-Galp, and (1→3,6)-Galp in a relative proportion of approximately 6:4:3:3:3, suggesting an arabinogalactan/galactoarabinan polysacchairde. With the aid of arabinan degrading enzymes, the structure of DAG1 repeating unit was further characterized by ELISA with specific monoclonal antibodies and Yariv reagent assay. Analyses indicated that the proposed repeating unit of DAG1 had a backbone composed of seven α-(1→5)-L-arabinofuranose residues where branching occurred at O-2 with either terminal arabinoses or arabinogalactan side chain. The arabinogalactan side chain was composed of six β-(1→3)-D-galactopyranose residues, half of which were ramified at O-6 with terminal arabinoses and the last galactose was terminated with arabinose. PMID:25498703

  19. Measuring phosphatidic acid phosphatase (EC 3.1.3.4) activity using two phosphomolybdate-based colorimetric methods

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phosphatidate phosphatase (3-sn-phosphatidate phosphohydrolase, EC 3.1.3.4), which is also known as PAP, catalyzes the dephosphorylation of phosphatidate (PtdOH) to form diacylglycerol (DAG) and inorganic phosphate. In eukaryotes, PAP driven reaction is the committed step in the synthesis of triacyl...

  20. 78 FR 69753 - Exemption of Records Systems Under the Privacy Act

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-21

    ... notice of proposed rulemaking (NPRM) reflecting these amendments in the Federal Register at 78 FR 56852.../OCDETF-001 at 78 FR 56737, Sept. 13, 2013, and for JUSTICE/OCDETF-002 at 78 FR 56926, Sept. 16, 2013...,'' JUSTICE/DAG-003, and the ] ``Organized Crime Drug Enforcement Task Force Fusion Center and...

  1. Identification, synthesis, and bioactivity of a male-produced aggregation pheromone in assassin bug,Pristhesancus Plagipennis (Hemiptera: Reduviidae).

    PubMed

    James, D G; Moore, C J; Aldrich, J R

    1994-12-01

    Pristhesancus plagipennis, a large Australian assassin bug, possesses three pairs of dorsal abdominal glands (DAGs). In the male, the anterior and posterior glands are hypertrophied and secrete an attractant pheromone. Gas chromatography-mass spectrometry (GC-MS) analyses of male DAG extracts and airborne volatiles emitted from calling males showed the pheromone signature to be dominated by a novel component. Subsequent chemical manipulations, GC-MS, and chiral-column analyses established its identity as (Z)-3-hexenyl (R)-2-hydroxy-3-methylbutyrate. Minor components included 3-methylbutanol, 2-phenylethanol, (Z)-3-hexenol, decanal, (E)-2-hexenoic acid, and three minor hexenyl esters. Bioactivity studies using laboratory olfactometers and outdoor flight cages demonstrated attraction by femaleP. plagipennis to calling males, heptane extracts of male posterior DAGs and a synthetic formulation of the (Z)R enantiomer of the major ester, alone or in combination with other components of male anterior and posterior DAGs. Males were also attracted to the major ester. The racemate andS enantiomer of the ester were not attractive. Contamination of the (Z)R enantiomer with 30-60% of theE isomer also made the compound nonattractive. This is the first report of an aggregation pheromone in the Reduviidae. The prospects for pheromonal manipulation ofP. plagipennis populations to enhance the value of this predator in horticultural ecosystems, are discussed. PMID:24241992

  2. Production of soy polyol oils: Results of microbial screening and identification of positive cultures

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Triacylglycerols (TAG) containing hydroxy fatty acids have many industrial uses, such as the manufacture of aviation lubricants, plastics, paints, nylons and cosmetics, because of the hydroxyl groups on the fatty acid (FA) constituents. Diacylglycerols (DAG) containing hydroxy FA can also be used in...

  3. Optimum conditions for the production of soy polyol oils and diacylglycerol from soybean oil by Acinetobacter haemolyticus A01-35 NRRL B-59985

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Triacylglycerols (TAG) containing hydroxy fatty acids have many industrial uses such as the manufacture of aviation lubricant, plastic, paint, nylons, and cosmetics, because of the hydroxyl groups on the fatty acid (FA) constituents. Diacylglycerols (DAG) containing hydroxy FA can also be used in th...

  4. Purification, characterization, and bioinformatics studies of phosphatidic acid phosphohydrolase from Lagenaria siceraria

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phosphatidic acid phosphohydrolase (PAP), EC 3.1.3.4, is the penultimate step in the Kennedy pathway of triacyl glycerol (TAG) synthesis leading to the formation of diacyl glycerol (DAG), which is a key intermediate in TAG synthesis. We partially purified a soluble PAP from mid maturing seeds of bot...

  5. Activation of protein kinase C by the lipid moieties of lipopolysaccharide

    SciTech Connect

    Wightman, P.D.; Raetz, C.R.H.

    1986-03-01

    Protein kinase C (PKC) was partially purified from the RAW264.7 macrophage-like cell and characterized by its activation by phosphatidylserine (PS) in the presence of calcium and its insensitivity to cyclic nucleotides or calmodulin. This PKC can also be activated by the acidic lipid moieties of lipopolysaccharide (LPS). The LPS lipids activate PKC in the absence of PS and, like PS, synergize with diacylglycerol (DAG). Intact RAW264.7 cells were prelabelled with /sup 32/Pi and treated with the well characterized PKC ligands, phorbol myristate acetate (PMA) or DAG. The phosphoproteins thereby induced were separated in 2-D gels and visualized by autoradiography. These phosphoproteins were used as standards to identify the PKC-associated phosphoproteins induced in these cells using other stimulators. The authors demonstrate that the LPS lipids as well as LPS itself induce the formation of phosphoproteins common to those induced by PMA or DAG. PMA, DAG, the LPS lipids, and LPS itself activate the RAW264.7 cell and stimulate the release of prostaglandin D/sub 2/ at the same concentration that stimulate new protein phosphorylation. These results suggest that the activation of PKC is an early event in the activation of the RAW264.7 macrophage by LPS.

  6. Thermal profiles, crystallization behaviors and microstructure of diacylglycerol-enriched palm oil blends with diacylglycerol-enriched palm olein.

    PubMed

    Xu, Yayuan; Zhao, Xiaoqing; Wang, Qiang; Peng, Zhen; Dong, Cao

    2016-07-01

    To elucidate the possible interaction mechanisms between DAG-enriched oils, this study investigated how mixtures of DAG-enriched palm-based oils influenced the phase behavior, thermal properties, crystallization behaviors and the microstructure in binary fat blends. DAG-enriched palm oil (PO-DAGE) was blended with DAG-enriched palm olein (POL-DAGE) in various percentages (0%, 10%, 30%, 50%, 70%, 90%, 100%). Based on the observation of iso-solid diagram and phase diagram, the binary mixture of PO-DAGE/POL-DAGE showed a better compatibility in comparison with their corresponding original blends. DSC thermal profiles exhibited that the melting and crystallization properties of PO-DAGE/POL-DAGE were distinctively different from corresponding original blends. Crystallization kinetics revealed that PO-DAGE/POL-DAGE blends displayed a rather high crystallization rate and exhibited no spherulitic crystal growth. From the results of polarized light micrographs, PO-DAGE/POL-DAGE blends showed more dense structure with very small needle-like crystals than PO/POL. X-ray diffraction evaluation revealed when POL-DAGE was added in high contents to PO-DAGE, above 30%, β-polymorph dominated, and the mount of β' forms crystals was decreasing. PMID:26920306

  7. Phosphatidic Acid-Mediated Signaling Regulates Microneme Secretion in Toxoplasma.

    PubMed

    Bullen, Hayley E; Jia, Yonggen; Yamaryo-Botté, Yoshiki; Bisio, Hugo; Zhang, Ou; Jemelin, Natacha Klages; Marq, Jean-Baptiste; Carruthers, Vern; Botté, Cyrille Y; Soldati-Favre, Dominique

    2016-03-01

    The obligate intracellular lifestyle of apicomplexan parasites necessitates an invasive phase underpinned by timely and spatially controlled secretion of apical organelles termed micronemes. In Toxoplasma gondii, extracellular potassium levels and other stimuli trigger a signaling cascade culminating in phosphoinositide-phospholipase C (PLC) activation, which generates the second messengers diacylglycerol (DAG) and IP3 and ultimately results in microneme secretion. Here we show that a delicate balance between DAG and its downstream product, phosphatidic acid (PA), is essential for controlling microneme release. Governing this balance is the apicomplexan-specific DAG-kinase-1, which interconverts PA and DAG, and whose depletion impairs egress and causes parasite death. Additionally, we identify an acylated pleckstrin-homology (PH) domain-containing protein (APH) on the microneme surface that senses PA during microneme secretion and is necessary for microneme exocytosis. As APH is conserved in Apicomplexa, these findings highlight a potentially widely used mechanism in which key lipid mediators regulate microneme exocytosis. PMID:26962945

  8. Stage-Specific Fatty Acid Fluxes Play a Regulatory Role in Glycerolipid Metabolism during Seed Development in Jatropha curcas L.

    PubMed

    Chaitanya, Bharatula Sri Krishna; Kumar, Sumit; Kaki, Shiva Shanker; Balakrishna, Marrapu; Karuna, Mallampalli Sri Lakshmi; Prasad, Rachapudi Badari Narayana; Sastry, Pidaparty Seshadri; Reddy, Attipalli Ramachandra

    2015-12-23

    The present study describes the changes in lipid profile as well as fatty acid fluxes during seed development in Jatropha curcas L. Endosperm from 34, 37, and 40 days after anthesis (DAA), incubated with [(14)C]acetate, showed significant synthesis of phosphatidylcholine (PC) at seed maturation. The fatty acid methyl ester profile showed PC from 34 DAA was rich in palmitic acid (16:0), whereas PC from 37 and 40 DAA was rich in oleic acid (18:1n-9). Molecular species analysis of diacylglycerol (DAG) indicated DAG (16:0/18:2n-6) was in abundance at 34 DAA, whereas DAG (18:1n-9/18:2n-6) was significantly high at 40 DAA. Triacylglycerol (TAG) analysis revealed TAG (16:0/18:2n-6/16:0) was abundant at 34 DAA, whereas TAG (18:1n-9/18:2n-6/18:1n-9) formed the majority at 40 DAA. Expression of two types of diacylglycerol acyltransferases varied with seed maturation. These data demonstrate stage-specific distinct pools of PC and DAG synthesis during storage TAG accumulation in Jatropha seed. PMID:26628196

  9. Indexing hierarchical structures using graph spectra.

    PubMed

    Shokoufandeh, Ali; Macrini, Diego; Dickinson, Sven; Siddiqi, Kaleem; Zucker, Steven W

    2005-07-01

    Hierarchical image structures are abundant in computer vision and have been used to encode part structure, scale spaces, and a variety of multiresolution features. In this paper, we describe a framework for indexing such representations that embeds the topological structure of a directed acyclic graph (DAG) into a low-dimensional vector space. Based on a novel spectral characterization of a DAG, this topological signature allows us to efficiently retrieve a promising set of candidates from a database of models using a simple nearest-neighbor search. We establish the insensitivity of the signature to minor perturbation of graph structure due to noise, occlusion, or node split/merge. To accommodate large-scale occlusion, the DAG rooted at each nonleaf node of the query "votes" for model objects that share that "part," effectively accumulating local evidence in a model DAG's topological subspaces. We demonstrate the approach with a series of indexing experiments in the domain of view-based 3D object recognition using shock graphs. PMID:16013759

  10. Correlation of diacylglycerol level and protein kinase C activity in rat retina to retinal circulation.

    PubMed

    Shiba, T; Inoguchi, T; Sportsman, J R; Heath, W F; Bursell, S; King, G L

    1993-11-01

    The increases in diacylglycerol (DAG) level and protein kinase C (PKC) activity have been characterized biochemically and functionally in the retina and the brain of diabetic rats as well as in cultured vascular cells. PKC specific activities were increased in the membraneous fraction of retina from streptozotocin (STZ)-induced diabetic rats and the genetically determined diabetic BB rats, respectively, after 1 or 2 wk of diabetes, compared with control. The ratio of total PKC activities from membraneous and cytosol fractions was also increased in the retina of diabetic rats. With diabetes, all the isoenzymes and the total DAG level were increased in the rat retina, whereas no changes were found in the rat brain. Insulin treatment normalized plasma glucose levels and partially prevented the increases in the membraneous PKC activity and all the isoenzymes in the retina. In the retinal endothelial cells, the total DAG level and PKC specific activities are increased by 36 and 22%, respectively, in the membraneous pool when the glucose levels are changed from 5.5 to 22 mM. Activation of PKC activity and isoform beta II by the vitreal injection of phorbol dibutyrate mimicked the abnormal retinal blood circulation observed in diabetic rats (2.22 +/- 0.24 vs. 1.83 +/- 0.40 s). Thus diabetes and elevated glucose levels will increase DAG level and PKC activities and its isoenzyme specifically in vascular cells and may affect retinal hemodynamics. PMID:8238505

  11. New eucrite Dar al Gani 872: Petrography, chemical composition, and evolution

    NASA Astrophysics Data System (ADS)

    Patzer, A.; Hill, D. H.; Boynton, W. V.

    2003-05-01

    Dar al Gani 872 (DaG 872) is a new meteorite from Libya that we classified by means of Instrumental Neutron Activation Analysis (INAA), electron microprobe, and optical microscopy. According to our results, DaG 872 is a Mg-rich main group eucrite, i.e., a monomict noncumulate basaltic eucrite displaying a predominant coarse-grained relict subophitic and a fine-grained granulitic texture. The meteorite also shows pockets of late-stage mesostasis and is penetrated by several calcite veins due to terrestrial weathering. Finally, it exhibits shock phenomena of stage 1­2 including heavily fractured mineral components, undulose extinction of plagioclase, kinked lamellae, and mosaicism in pyroxenes corresponding to peak pressures of ~20 GPa. In view of petrographic criteria as well as compositional and exsolution characteristics of its pyroxenes, the sample represents a metamorphic type 5 eucrite. Assuming the metamorphic type to be a function of burial depth on the parent body and taking into account the relatively high shock stage, the excavation of DaG 872 was likely induced by a major impact event. Prior to this point, DaG 872 apparently underwent a 4-stage geological evolution that is reflected by intricate textural and mineralogical features.

  12. Studying Intrapersonal Communication through Memoirs and Journals.

    ERIC Educational Resources Information Center

    Jensen, Marvin D.

    One theoretical approach to self-awareness is the study of introspective writing. Among the characteristics of intrapersonal communication that can be explored through memoirs and journals are selective memory and chronology revision. From the reflections of writers like Lillian Hellman, Dag Hammarskjold, and May Sarton comes confirmation that…

  13. Proliferating or Differentiating Stimuli Act on Different Lipid-dependent Signaling Pathways in Nuclei of Human Leukemia Cells

    PubMed Central

    Neri, Luca M.; Bortul, Roberta; Borgatti, Paola; Tabellini, Giovanna; Baldini, Giovanna; Capitani, Silvano; Martelli, Alberto M.

    2002-01-01

    Previous results have shown that the human promyelocytic leukemia HL-60 cell line responds to either proliferating or differentiating stimuli. When these cells are induced to proliferate, protein kinase C (PKC)-βII migrates toward the nucleus, whereas when they are exposed to differentiating agents, there is a nuclear translocation of the α isoform of PKC. As a step toward the elucidation of the early intranuclear events that regulate the proliferation or the differentiation process, we show that in the HL-60 cells, a proliferating stimulus (i.e., insulin-like growth factor-I [IGF-I]) increased nuclear diacylglycerol (DAG) production derived from phosphatidylinositol (4,5) bisphosphate, as indicated by the inhibition exerted by 1-O-octadeyl-2-O-methyl-sn-glycero-3-phosphocholine and U-73122 (1-[6((17β-3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl]-1H-pyrrole-2,5-dione), which are pharmacological inhibitors of phosphoinositide-specific phospholipase C. In contrast, when HL-60 cells were induced to differentiate along the granulocytic lineage by dimethyl sulfoxide, we observed a rise in the nuclear DAG mass, which was sensitive to either neomycin or propranolol, two compounds with inhibitory effect on phospholipase D (PLD)-mediated DAG generation. In nuclei of dimethyl sulfoxide-treated HL-60 cells, we observed a rise in the amount of a 90-kDa PLD, distinct from PLD1 or PLD2. When a phosphatidylinositol (4,5) bisphosphate-derived DAG pool was generated in the nucleus, a selective translocation of PKC-βII occurred. On the other hand, nuclear DAG derived through PLD, recruited PKC-α to the nucleus. Both of these PKC isoforms were phosphorylated on serine residues. These results provide support for the proposal that in the HL-60 cell nucleus there are two independently regulated sources of DAG, both of which are capable of acting as the driving force that attracts to this organelle distinct, DAG-dependent PKC isozymes. Our results assume a particular

  14. Role of phospholipase D and diacylglycerol in activating constitutive TRPC-like cation channels in rabbit ear artery myocytes.

    PubMed

    Albert, A P; Piper, A S; Large, W A

    2005-08-01

    Previously we have described a constitutively active Ca2+-permeable non-selective cation channel in freshly dispersed rabbit ear artery myocytes that has similar properties to canonical transient receptor potential (TRPC) channel proteins. In the present study we have investigated the transduction pathways responsible for stimulating constitutive channel activity in these myocytes. Application of the pharmacological inhibitors of phosphatidylcholine-phospholipase D (PC-PLD), butan-1-ol and C2 ceramide, produced marked inhibition of constitutive channel activity in cell-attached patches and also butan-1-ol produced pronounced suppression of resting membrane conductance measured with whole-cell recording whereas the inactive isomer butan-2-ol had no effect on constitutive whole-cell or channel activity. In addition butan-1-ol had no effect on channel activity evoked by the diacylglycerol (DAG) analogue 1-oleoyl-2-acetyl-sn-glycerol (OAG). Inhibitors of PC-phospholipase C (PC-PLC) and phospholipase A2 (PLA2) had no effect on constitutive channel activity. Application of a purified PC-PLD enzyme and its metabolite phosphatidic acid to inside-out patches markedly increased channel activity. The phosphatidic acid phosphohydrolase (PAP) inhibitor dl-propranolol also inhibited constitutive and phosphatidic acid-induced increases in channel activity but had no effect on OAG-evoked responses. The DAG lipase and DAG kinase inhibitors, RHC80267 and R59949 respectively, which inhibit DAG metabolism, produced transient increases in channel activity which were mimicked by relatively high concentrations (40 microm) of OAG. The protein kinase C (PKC) inhibitor chelerythrine did not prevent channel activation by OAG but blocked the secondary inhibitory response of OAG. It is proposed that endogenous DAG is involved in the activation of channel activity and that its effects on channel activity are concentration-dependent with higher concentrations of DAG also inhibiting channel

  15. Stimulus-response coupling in insulin-secreting HIT cells. Effects of secretagogues on cytosolic Ca2+, diacylglycerol, and protein kinase C activity.

    PubMed

    Regazzi, R; Li, G D; Deshusses, J; Wollheim, C B

    1990-09-01

    The hamster islet B cell line HIT retains the ability to secret insulin in response to glucose and several receptor agonists. We used HIT cells to study the initial signaling events in glucose or receptor agonist-stimulated insulin secretion. Glucose stimulated insulin release from HIT cells in a dose-dependent manner with a half-maximal effect seen already at 1 mM. Insulin release was also stimulated by carbachol in a glucose-dependent manner. Glucose depolarized the HIT cell membrane potential as assessed with the fluorescent probe bisoxonol and raised intracellular Ca2+ as revealed by fura-2 measurements. Using a Mn2+ fura-2 quenching technique, we could show that the rise in intracellular Ca2+ was due to Ca2+ influx following opening of voltage-gated Ca2+ channels. Glucose is thought to increase the diacylglycerol (DAG) content of insulin-secreting cells. However, although HIT cells respond to glucose in terms of insulin secretion, membrane depolarization, and Ca2+ rise, the hexose was unable to increase the proportion of protein kinase C activity associated with membranes. In contrast, the membrane-associated protein kinase C activity increased in HIT cells exposed to the two receptor agonists carbachol and bombesin. Bombesin was shown to generate DAG with the expected fatty acid composition of activators of phospholipase C. Glucose, in contrast, only caused minor increases in DAG containing myristic and palmitic acid without affecting total DAG mass. The failure to detect stimulation of protein kinase C by glucose could be due to both the limited amount and to the different fatty acid composition of the metabolically generated DAG. The latter was in part supported by experiments performed on protein kinase C partially purified from HIT cells. Indeed, 1,2-dipalmitoylglycerol, presumed to be the main DAG species generated by glucose, was only one-third as active as 1,2-dioleoylglycerol and 1-stearoyl-2-arachidonylglycerol in stimulating the isolated enzyme at

  16. Neutron diffraction study of the monoclinic to tetragonal structural transition in LaNbO{sub 4} and its relation to proton mobility

    SciTech Connect

    Huse, M.; Skilbred, A.W.B.; Karlsson, M.; Eriksson, S.G.; Norby, T.; Haugsrud, R.; Knee, C.S.

    2012-03-15

    The fergusonite-scheelite structural transition of LaNbO{sub 4} was characterized by high-intensity variable temperature neutron powder diffraction in an effort to link structural changes to proton transport in the low temperature monoclinic and high temperature tetragonal polymorphs. The pronounced decrease in enthalpy of proton mobility with increasing temperature in monoclinic LaNbO{sub 4}, identified by Fjeld et al. , was found to coincide with the decrease in a key inter-tetrahedral oxygen separation. At temperatures above the transition, this oxygen to oxygen distance remains constant, behavior that is consistent with the invariant nature of the calculated enthalpy of mobility for the tetragonal phase. This oxygen separation is therefore proposed as the key structural parameter determining the success rate of proton transfer and ultimately the proton diffusion in LaNbO{sub 4}. Infrared measurements performed on acceptor-doped LaNbO{sub 4} show an absorption in the region 2500 to 3700 cm{sup -1} attributed to the O-H stretching motion influenced by hydrogen bonding. - Graphical Abstract: The structural phase transition from monoclinic fergusonite to tetragonal scheelite crystal structure clearly influences the proton conductivity of acceptor-doped LaNbO{sub 4} (see Figure). The present article attempts to explain why. Highlights: Black-Right-Pointing-Pointer Transition from monoclinic to tetragonal structure studied using neutron diffraction. Black-Right-Pointing-Pointer Change in proton mobility linked to contraction of a key oxygen separation. Black-Right-Pointing-Pointer Infra-red spectra reveal intermediate strength hydrogen bonding.

  17. Solvent-Free Lipase-Catalyzed Synthesis of Diacylgycerols as Low-Calorie Food Ingredients.

    PubMed

    Vázquez, Luis; González, Noemí; Reglero, Guillermo; Torres, Carlos

    2016-01-01

    Problems derived from obesity and overweight have recently promoted the development of fat substitutes and other low-calorie foods. On the one hand, fats with short- and medium-chain fatty acids are a source of quick energy, easily hydrolyzable and hardly stored as fat. Furthermore, 1,3-diacylglycerols are not hydrolyzed to 2-monoacylglycerols in the gastrointestinal tract, reducing the formation of chylomicron and lowers the serum level of triacylglycerols by decreasing its resynthesis in the enterocyte. In this work, these two effects were combined to synthesize short- and medium-chain 1,3-diacylglycerols, leading to a product with great potential as for their low-calorie properties. Lipase-catalyzed transesterification reactions were performed between short- and medium-chain fatty acid ethyl esters and glycerol. Different variables were investigated, such as the type of biocatalyst, the molar ratio FAEE:glycerol, the adsorption of glycerol on silica gel, or the addition of lecithin. Best reaction conditions were evaluated considering the percentage of 1,3-DAG produced and the reaction rate. Except Novozym 435 (Candida antarctica), other lipases required the adsorption of glycerol on silica gel to form acylglycerols. Lipases that gave the best results with adsorption were Novozym 435 and Lipozyme RM IM (Rhizomucor miehei) with 52 and 60.7% DAG at 32 h, respectively. Because of its specificity for sn-1 and sn-3 positions, lipases leading to a higher proportion of 1,3-DAG vs. 1,2-DAG were Lipozyme RM IM (39.8 and 20.9%, respectively) and Lipase PLG (Alcaligenes sp.) (35.9 and 19.3%, respectively). By adding 1% (w/w) of lecithin to the reaction with Novozym 435 and raw glycerol, the reaction rate was considerably increased from 41.7 to 52.8% DAG at 24 h. PMID:26904539

  18. The effect of CD137-CD137 ligand interaction on phospholipase C signaling pathway in human endothelial cells.

    PubMed

    Yan, Jinchuan; Wang, Cuiping; Wang, Zhongqun; Yuan, Wei

    2013-11-25

    We previously reported the emerging role of CD137-CD137L interaction in inflammation and atherosclerosis. The mechanism of CD137-CD137L interaction may be related to a variety of signaling pathways. The most important signaling pathway involves the activation of phospholipase C(PLC) which induces the diacylglycerol-protein kinase C(DAG-PKC) and the inositol trisphosphate-intracellular free calcium (IP3-[Ca(2+)]i) pathway. In the current study, we investigated whether CD137-CD137L interaction can stimulate the PLC signaling pathway in human umbilical vein endothelial cells (HUVEC). The diacylglycerol (DAG) and inositol trisphosphate (IP3) levels in HUVEC were measured by radioenzymatic assay. The activity of protein kinase (PKC) was detected by its ability to transfer phosphate from [γ-(32)P]ATP to lysine-rich histone. The [Ca(2+)]i concentrations were measured by flow cytometric analysis. The DAG level and PKC activity were increased in a concentration-dependent, biphasic manner in HUVEC induced by anti-CD137. PKC activity was mainly in the cytosol at rest, and then translocated to the membrane when stimulated by anti-CD137. Similarly, rapid IP3 formation induced by anti-CD137 coincided with the peak of the DAG level. Moreover, anti-CD137 induced peak [Ca(2+)]i responses including the rapid transient phase and the sustained phase. However, anti-CD137L suppressed the activation of the DAG-PKC and IP3-[Ca(2+)]i signaling pathway, which was stimulated by anti-CD137 in HUVEC. In conclusion, the data suggested that CD137-CD137L interaction induces robust activation of the PLC signaling pathway in HUVEC. PMID:24070733

  19. Subcellular localization of monoglucosyldiacylglycerol synthase in Synechocystis sp. PCC6803 and its unique regulation by lipid environment.

    PubMed

    Selão, Tiago Toscano; Zhang, Lifang; Ariöz, Candan; Wieslander, Åke; Norling, Birgitta

    2014-01-01

    Synthesis of monogalactosyldiacylglycerol (GalDAG) and digalactosyldiacylglycerol (GalGalDAG), the major membrane lipids in cyanobacteria, begins with production of the intermediate precursor monoglucosyldiacylglycerol (GlcDAG), by monoglucosyldiacylglycerol synthase (MGS). In Synechocystis sp. PCC6803 (Synechocystis) this activity is catalyzed by an integral membrane protein, Sll1377 or MgdA. In silico sequence analysis revealed that cyanobacterial homologues of MgdA are highly conserved and comprise a distinct group of lipid glycosyltransferases. Global regulation of lipid synthesis in Synechocystis and, more specifically, the influence of the lipid environment on MgdA activity have not yet been fully elucidated. Therefore, we purified membrane subfractions from this organism and assayed MGS activity in vitro, with and without different lipids and other potential effectors. Sulfoquinovosyldiacylglycerol (SQDAG) potently stimulates MgdA activity, in contrast to other enzymes of a similar nature, which are activated by phosphatidylglycerol instead. Moreover, the final products of galactolipid synthesis, GalDAG and GalGalDAG, inhibited this activity. Western blotting revealed the presence of MgdA both in plasma and thylakoid membranes, with a high specific level of the MgdA protein in the plasma membrane but highest MGS activity in the thylakoid membrane. This discrepancy in the subcellular localization of enzyme activity and protein may indicate the presence of either an unknown regulator and/or an as yet unidentified MGS-type enzyme. Furthermore, the stimulation of MgdA activity by SQDAG observed here provides a new insight into regulation of the biogenesis of both sulfolipids and galactolipids in cyanobacteria. PMID:24516600

  20. Building a Hierarchical Organization of Protein Complexes Out of Protein Association Data

    PubMed Central

    Stojmirović, Aleksandar; Yu, Yi-Kuo

    2014-01-01

    Organizing experimentally determined protein associations as a hierarchy can be a good approach to elucidating the content of protein complexes and the modularity of subcomplexes. Several challenges exist. First, intrinsically sticky proteins, such as chaperones, are often falsely assigned to many functionally unrelated complexes. Second, the reported collections of proteins may not be true “complexes” in the sense that they bind together and perform a joint cellular function. Third, due to imperfect sensitivity of protein detection methods, both false positive and false negative assignments of a protein to complexes may occur. We mitigate the first issue by down-weighting sticky proteins by their occurrence frequencies. We approach the other two problems by merging nearly identical complexes and by constructing a directed acyclic graph (DAG) based on the relationship of partial inclusion. The constructed DAG, within which smaller complexes form parts of the larger, can reveal how different complexes are joined. By merging almost identical complexes one can deemphasize the influence of false positives, while allowing false negatives to be rescued by other nearly identical association data. We investigate several protein weighting schemes and compare their corresponding DAGs using yeast and human complexes. We find that the scheme incorporating weights based on information flow in the network of direct protein–protein interactions produces biologically most meaningful DAGs. In either yeast or human, isolated nodes form a large proportion of the final hierarchy. While most connected components encompass very few nodes, the largest one for each species contains a sizable portion of all nodes. By considering examples of subgraphs composed of nodes containing a specified protein, we illustrate that the graphs' topological features can correctly suggest the biological roles of protein complexes. The input data, final results and the source code are available at ftp

  1. Diacylglycerol kinase α exacerbates cardiac injury after ischemia/reperfusion.

    PubMed

    Sasaki, Toshiki; Shishido, Tetsuro; Kadowaki, Shinpei; Kitahara, Tatsuro; Suzuki, Satoshi; Katoh, Shigehiko; Funayama, Akira; Netsu, Shunsuke; Watanabe, Tetsu; Goto, Kaoru; Takeishi, Yasuchika; Kubota, Isao

    2014-01-01

    Early coronary reperfusion of the ischemic myocardium is a desired therapeutic goal for the preservation of myocardial function. However, reperfusion itself causes additional myocardium injuries. Activation of the diacylglycerol-protein kinase C (DAG-PKC) cascade has been implicated in the cardioprotective effects occurring after ischemia/reperfusion (I/R). DAG kinase (DGK) controls cellular DAG levels by converting DAG to phosphatidic acid, and may act as an endogenous regulator of DAG-PKC signaling. In the present study, we examined the functional role of DGKα in cardiac injury after I/R in in vivo mouse hearts. We generated transgenic mice with cardiac-specific overexpression of DGKα (DGKα-TG). The left anterior descending coronary artery was transiently occluded for 20 min and reperfused for 24 h in DGKα-TG mice and wild-type littermate (WT) mice. The levels of phosphorylation activity of PKCε, extracellular-signal regulated kinase (ERK) 1/2, and p70 ribosomal S6 kinase (p70S6K) were increased after I/R in WT mouse hearts. However, in DGKα-TG mice, activation of PKCε, ERK1/2, and p70S6K was attenuated compared to WT mice. After 24 h, Evans blue/triphenyltetrazolium chloride double staining and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining showed that DGKα-TG mice had significantly larger myocardial infarctions and larger numbers of TUNEL-positive cardiomyocytes than WT mice. Echocardiography and cardiac catheterization revealed that left ventricular systolic function was more severely depressed in DGKα-TG mice than in WT mice after I/R. These findings suggest that DGKα exacerbates I/R injury by inhibiting the cardioprotective effects of PKCε, ERK1/2, and p70S6K activation. PMID:23719772

  2. The opposing effects of ghrelin on hypothalamic and systemic inflammatory processes are modulated by its acylation status and food intake in male rats.

    PubMed

    García-Cáceres, Cristina; Fuente-Martín, Esther; Díaz, Francisca; Granado, Miriam; Argente-Arizón, Pilar; Frago, Laura M; Freire-Regatillo, Alejandra; Barrios, Vicente; Argente, Jesús; Chowen, Julie A

    2014-08-01

    Ghrelin is an endogenous hormone that stimulates appetite and adipose tissue accrual. Both the acylated (AG) and non-acylated (DAG) isoforms of this hormone are also reported to exert anti-inflammatory and protective effects systemically and in the central nervous system. As inflammatory processes have been implicated in obesity-associated secondary complications, we hypothesized that this natural appetite stimulator may protect against negative consequences resulting from excessive food intake. Adult male Wistar rats were treated icv (5 μg/day) with AG, DAG, the ghrelin mimetic GH-releasing peptide (GHRP)-6, AG, and pair-fed with controls (AG-pf) or saline for 14 days. Regardless of food intake AG increased visceral adipose tissue (VAT) and decreased circulating cytokine levels. However, AG reduced cytokine production in VAT only in rats fed ad libitum. Hypothalamic cytokine production was increased in AG-treated rats fed ad libitum and by DAG, but intracellular inflammatory signaling pathways associated with insulin and leptin resistance were unaffected. Gliosis was not observed in response to any treatment as glial markers were either reduced or unaffected. AG, DAG, and GHRP-6 stimulated production of hypothalamic insulin like-growth factor I that is involved in cell protective mechanisms. In hypothalamic astrocyte cell cultures AG decreased tumor necrosis factorα and DAG decreased interleukin-1β mRNA levels, suggesting direct anti-inflammatory effects on astrocytes. Thus, whereas ghrelin stimulates food intake and weight gain, it may also induce mechanisms of cell protection that help to detour or delay systemic inflammatory responses and hypothalamic gliosis due to excess weight gain, as well as its associated pathologies. PMID:24848869

  3. Preferential elevation of protein kinase C isoform beta II and diacylglycerol levels in the aorta and heart of diabetic rats: differential reversibility to glycemic control by islet cell transplantation.

    PubMed Central

    Inoguchi, T; Battan, R; Handler, E; Sportsman, J R; Heath, W; King, G L

    1992-01-01

    In the present study, we have measured protein kinase C (PKC) specific activities and total diacylglycerol (DAG) level in the aorta and heart of rats, which showed that after 2 weeks of streptozotocin (STZ)-induced diabetes, membranous PKC specific activity and total DAG content were increased significantly by 88% and 40% in the aorta and by 21% and 72% in the heart, respectively. Hyperglycemia was identified as being a causal factor since elevated glucose levels increased DAG levels in cultured aortic endothelial and smooth muscle cells. Analysis by immunoblotting revealed that only alpha and beta II PKC isoenzymes are detected in these two tissues and vascular cells among those studied. In STZ-induced diabetic rats, beta II isoenzyme is preferentially increased in both aorta and heart, whereas PKC alpha did not change significantly. The increases in membranous PKC specific activity and DAG level are observed in both spontaneous diabetes-prone diabetic BB rats as well as in STZ-induced diabetic BB and Sprague-Dawley rats, which persisted for up to 5 weeks. After 2 weeks of diabetes without treatment, the normalization of blood glucose levels for up to 3 weeks with islet cell transplants in STZ-induced diabetic BB rats reversed the biochemical changes only in the heart, but not in the aorta. These results suggest that PKC activity and DAG level may be persistently activated in the macrovascular tissues from diabetic animals and indicate a possible role for these biochemical parameters in the development of diabetic chronic vascular complications. Images PMID:1438315

  4. Solvent-Free Lipase-Catalyzed Synthesis of Diacylgycerols as Low-Calorie Food Ingredients

    PubMed Central

    Vázquez, Luis; González, Noemí; Reglero, Guillermo; Torres, Carlos

    2016-01-01

    Problems derived from obesity and overweight have recently promoted the development of fat substitutes and other low-calorie foods. On the one hand, fats with short- and medium-chain fatty acids are a source of quick energy, easily hydrolyzable and hardly stored as fat. Furthermore, 1,3-diacylglycerols are not hydrolyzed to 2-monoacylglycerols in the gastrointestinal tract, reducing the formation of chylomicron and lowers the serum level of triacylglycerols by decreasing its resynthesis in the enterocyte. In this work, these two effects were combined to synthesize short- and medium-chain 1,3-diacylglycerols, leading to a product with great potential as for their low-calorie properties. Lipase-catalyzed transesterification reactions were performed between short- and medium-chain fatty acid ethyl esters and glycerol. Different variables were investigated, such as the type of biocatalyst, the molar ratio FAEE:glycerol, the adsorption of glycerol on silica gel, or the addition of lecithin. Best reaction conditions were evaluated considering the percentage of 1,3-DAG produced and the reaction rate. Except Novozym 435 (Candida antarctica), other lipases required the adsorption of glycerol on silica gel to form acylglycerols. Lipases that gave the best results with adsorption were Novozym 435 and Lipozyme RM IM (Rhizomucor miehei) with 52 and 60.7% DAG at 32 h, respectively. Because of its specificity for sn-1 and sn-3 positions, lipases leading to a higher proportion of 1,3-DAG vs. 1,2-DAG were Lipozyme RM IM (39.8 and 20.9%, respectively) and Lipase PLG (Alcaligenes sp.) (35.9 and 19.3%, respectively). By adding 1% (w/w) of lecithin to the reaction with Novozym 435 and raw glycerol, the reaction rate was considerably increased from 41.7 to 52.8% DAG at 24 h. PMID:26904539

  5. Influence of Gender, Obesity, and Muscle Lipase Activity on Intramyocellular Lipids in Sedentary Individuals

    PubMed Central

    Moro, Cedric; Galgani, Jose E.; Luu, LanChi; Pasarica, Magdalena; Mairal, Aline; Bajpeyi, Sudip; Schmitz, Gerd; Langin, Dominique; Liebisch, Gerhard; Smith, Steven R.

    2009-01-01

    Context: Obesity and type 2 diabetes are associated with elevated intramyocellular lipids (IMCLs) and insulin resistance. Objective: We tested the hypothesis that skeletal muscle lipases activity could influence IMCL content (including diacylglycerol and ceramides). Design and Patients: The present study included 48 subjects with a wide range of age (19–68 yr) and body mass index (20–45 kg/m2) who underwent skeletal muscle biopsy, dual-energy x-ray absorptiometry and a hyperinsulinemic euglycemic clamp. Main Outcome Measures: Insulin sensitivity by hyperinsulinemic clamp, and intramyocellular triacylglycerol (IMTG), diacylglycerol (DAG), and ceramides content, and triacylglycerol and diacylglycerol hydrolase activities were measured in biopsies of vastus lateralis. IMCL was measured by 1H-magnetic resonance spectroscopy in a subgroup of 25 subjects. Multivariate regression analyses were performed to identify the main predictors of IMCL. Results: Body fat was the main predictor of IMTG independently of the method and the type of muscle; IMTG concentration was higher in females vs. males and obese vs. nonobese subjects. Muscle DAG and ceramides concentrations were elevated in obese and type 2 diabetic subjects and were not related to body fat and fasting free fatty acids, whereas a direct association with the ratio of diacylglycerol hydrolase to triacylglycerol hydrolase activity (an index of incomplete triacylglycerol hydrolysis) was observed, which explained 54 and 38% of the variance in DAG and ceramides (P < 0.001), respectively. DAG content was the main determinant of insulin resistance. Conclusions: These data suggest that intramyocellular DAG is an independent predictor of insulin resistance in humans and that its levels correlate with lipolytic enzymes activity in skeletal muscle but not with markers of adiposity. PMID:19531593

  6. Preferential elevation of protein kinase C isoform beta II and diacylglycerol levels in the aorta and heart of diabetic rats: differential reversibility to glycemic control by islet cell transplantation.

    PubMed

    Inoguchi, T; Battan, R; Handler, E; Sportsman, J R; Heath, W; King, G L

    1992-11-15

    In the present study, we have measured protein kinase C (PKC) specific activities and total diacylglycerol (DAG) level in the aorta and heart of rats, which showed that after 2 weeks of streptozotocin (STZ)-induced diabetes, membranous PKC specific activity and total DAG content were increased significantly by 88% and 40% in the aorta and by 21% and 72% in the heart, respectively. Hyperglycemia was identified as being a causal factor since elevated glucose levels increased DAG levels in cultured aortic endothelial and smooth muscle cells. Analysis by immunoblotting revealed that only alpha and beta II PKC isoenzymes are detected in these two tissues and vascular cells among those studied. In STZ-induced diabetic rats, beta II isoenzyme is preferentially increased in both aorta and heart, whereas PKC alpha did not change significantly. The increases in membranous PKC specific activity and DAG level are observed in both spontaneous diabetes-prone diabetic BB rats as well as in STZ-induced diabetic BB and Sprague-Dawley rats, which persisted for up to 5 weeks. After 2 weeks of diabetes without treatment, the normalization of blood glucose levels for up to 3 weeks with islet cell transplants in STZ-induced diabetic BB rats reversed the biochemical changes only in the heart, but not in the aorta. These results suggest that PKC activity and DAG level may be persistently activated in the macrovascular tissues from diabetic animals and indicate a possible role for these biochemical parameters in the development of diabetic chronic vascular complications. PMID:1438315

  7. Caloric restriction and intermittent fasting alter hepatic lipid droplet proteome and diacylglycerol species and prevent diabetes in NZO mice.

    PubMed

    Baumeier, Christian; Kaiser, Daniel; Heeren, Jörg; Scheja, Ludger; John, Clara; Weise, Christoph; Eravci, Murat; Lagerpusch, Merit; Schulze, Gunnar; Joost, Hans-Georg; Schwenk, Robert Wolfgang; Schürmann, Annette

    2015-05-01

    Caloric restriction and intermittent fasting are known to improve glucose homeostasis and insulin resistance in several species including humans. The aim of this study was to unravel potential mechanisms by which these interventions improve insulin sensitivity and protect from type 2 diabetes. Diabetes-susceptible New Zealand Obese mice were either 10% calorie restricted (CR) or fasted every other day (IF), and compared to ad libitum (AL) fed control mice. AL mice showed a diabetes prevalence of 43%, whereas mice under CR and IF were completely protected against hyperglycemia. Proteomic analysis of hepatic lipid droplets revealed significantly higher levels of PSMD9 (co-activator Bridge-1), MIF (macrophage migration inhibitor factor), TCEB2 (transcription elongation factor B (SIII), polypeptide 2), ACY1 (aminoacylase 1) and FABP5 (fatty acid binding protein 5), and a marked reduction of GSTA3 (glutathione S-transferase alpha 3) in samples of CR and IF mice. In addition, accumulation of diacylglycerols (DAGs) was significantly reduced in livers of IF mice (P=0.045) while CR mice showed a similar tendency (P=0.062). In particular, 9 DAG species were significantly reduced in response to IF, of which DAG-40:4 and DAG-40:7 also showed significant effects after CR. This was associated with a decreased PKCε activation and might explain the improved insulin sensitivity. In conclusion, our data indicate that protection against diabetes upon caloric restriction and intermittent fasting associates with a modulation of lipid droplet protein composition and reduction of intracellular DAG species. PMID:25645620

  8. Modal analysis of jointed structures

    NASA Astrophysics Data System (ADS)

    Quinn, D. Dane

    2012-01-01

    Structural systems are often composed of multiple components joined together at localized interfaces. Compared to a corresponding monolithic system these interfaces are designed to have little influence on the load carrying capability of the system, and the resulting change in the overall system mass and stiffness is minimal. Hence, under nominal operating conditions the mode shapes and frequencies of the dominant structural modes are relatively insensitive to the presence of the interfaces. However, the energy dissipation in such systems is strongly dependent on the joints. The microslip that occurs at each interface couples together the structural modes of the system and introduces nonlinear damping into the system, effectively altering the observed damping of the structural modes, which can then significantly alter the amplitude of the response at the resonant modal frequencies. This work develops equations of motion for a jointed structure in terms of the structural modal coordinates and implements a reduced-order description of the microslip that occurs at the interface between components. The interface is incorporated into the modal description of the system through an existing decomposition of a series-series Iwan interface model and a continuum approximation for microslip of an elastic rod. The developed framework is illustrated on several examples, including a discrete three degree-of-freedom system as well as the longitudinal deformation of a continuum beam.

  9. Ontologies and tag-statistics

    NASA Astrophysics Data System (ADS)

    Tibély, Gergely; Pollner, Péter; Vicsek, Tamás; Palla, Gergely

    2012-05-01

    Due to the increasing popularity of collaborative tagging systems, the research on tagged networks, hypergraphs, ontologies, folksonomies and other related concepts is becoming an important interdisciplinary area with great potential and relevance for practical applications. In most collaborative tagging systems the tagging by the users is completely ‘flat’, while in some cases they are allowed to define a shallow hierarchy for their own tags. However, usually no overall hierarchical organization of the tags is given, and one of the interesting challenges of this area is to provide an algorithm generating the ontology of the tags from the available data. In contrast, there are also other types of tagged networks available for research, where the tags are already organized into a directed acyclic graph (DAG), encapsulating the ‘is a sub-category of’ type of hierarchy between each other. In this paper, we study how this DAG affects the statistical distribution of tags on the nodes marked by the tags in various real networks. The motivation for this research was the fact that understanding the tagging based on a known hierarchy can help in revealing the hidden hierarchy of tags in collaborative tagging systems. We analyse the relation between the tag-frequency and the position of the tag in the DAG in two large sub-networks of the English Wikipedia and a protein-protein interaction network. We also study the tag co-occurrence statistics by introducing a two-dimensional (2D) tag-distance distribution preserving both the difference in the levels and the absolute distance in the DAG for the co-occurring pairs of tags. Our most interesting finding is that the local relevance of tags in the DAG (i.e. their rank or significance as characterized by, e.g., the length of the branches starting from them) is much more important than their global distance from the root. Furthermore, we also introduce a simple tagging model based on random walks on the DAG, capable of

  10. Cosmogenic Records in 18 Ordinary Chondrites from the Dar Al Gani Region, Libya. 1; Noble Gases

    NASA Technical Reports Server (NTRS)

    Schultz, L.; Franke, L.; Welten, K. C.; Nishiizumi, K.; Jull, A. J. T.

    2003-01-01

    In the last decade thousands of meteorites have been recovered from hot deserts in the Sahara and Oman. One of the main meteorite concentration surfaces in the Sahara is the Dar al Gani plateau in Libya, which covers a total area of 8000 km2. More than 1000 meteorites have been reported from this area. The geological setting, meteorite pairings and the meteorite density of the Dar al Gani (DaG) field are described in more detail in [1]. In this work we report concentrations of the noble gas isotopes of He, Ne, Ar as well as 84Kr and 132Xe in 18 DaG meteorites. In a separate paper we will report the cosmogenic radionuclides [2]. We discuss the thermal history and cosmic-ray exposure (CRE) history of these meteorites, and evaluate the effects of the hot desert environment on the noble gas record.

  11. Mammalian phospholipase C.

    PubMed

    Kadamur, Ganesh; Ross, Elliott M

    2013-01-01

    Phospholipase C (PLC) converts phosphatidylinositol 4,5-bisphosphate (PIP(2)) to inositol 1,4,5-trisphosphate (IP(3)) and diacylglycerol (DAG). DAG and IP(3) each control diverse cellular processes and are also substrates for synthesis of other important signaling molecules. PLC is thus central to many important interlocking regulatory networks. Mammals express six families of PLCs, each with both unique and overlapping controls over expression and subcellular distribution. Each PLC also responds acutely to its own spectrum of activators that includes heterotrimeric G protein subunits, protein tyrosine kinases, small G proteins, Ca(2+), and phospholipids. Mammalian PLCs are autoinhibited by a region in the catalytic TIM barrel domain that is the target of much of their acute regulation. In combination, the PLCs act as a signaling nexus that integrates numerous signaling inputs, critically governs PIP(2) levels, and regulates production of important second messengers to determine cell behavior over the millisecond to hour timescale. PMID:23140367

  12. Insights on TRP Channels from In Vivo Studies in Drosophila

    PubMed Central

    Minke, Baruch; Parnas, Moshe

    2007-01-01

    Transient receptor potential (TRP) channels mediate responses in a large variety of signaling mechanisms. Most studies on mammalian TRP channels rely on heterologous expression, but their relevance to in vivo tissues is not entirely clear. In contrast, Drosophila TRP and TRP-like (TRPL) channels allow direct analyses of in vivo function. In Drosophila photoreceptors, activation of TRP and TRPL is mediated via the phosphoinositide cascade, with both Ca2+ and diacylglycerol (DAG) essential for generating the light response. In tissue culture cells, TRPL channels are constitutively active, and lipid second messengers greatly facilitate this activity. Inhibition of phospholipase C (PLC) completely blocks lipid activation of TRPL, suggesting that lipid activation is mediated via PLC. In vivo studies in mutant Drosophila also reveal an acute requirement for lipid-producing enzyme, which may regulate PLC activity. Thus, PLC and its downstream second messengers, Ca2+ and DAG, constitute critical mediators of TRP/TRPL gating in vivo. PMID:16460287

  13. Correlative and integrated light and electron microscopy of in-resin GFP fluorescence, used to localise diacylglycerol in mammalian cells.

    PubMed

    Peddie, Christopher J; Blight, Ken; Wilson, Emma; Melia, Charlotte; Marrison, Jo; Carzaniga, Raffaella; Domart, Marie-Charlotte; O'Toole, Peter; Larijani, Banafshe; Collinson, Lucy M

    2014-08-01

    Fluorescence microscopy of GFP-tagged proteins is a fundamental tool in cell biology, but without seeing the structure of the surrounding cellular space, functional information can be lost. Here we present a protocol that preserves GFP and mCherry fluorescence in mammalian cells embedded in resin with electron contrast to reveal cellular ultrastructure. Ultrathin in-resin fluorescence (IRF) sections were imaged simultaneously for fluorescence and electron signals in an integrated light and scanning electron microscope. We show, for the first time, that GFP is stable and active in resin sections in vacuo. We applied our protocol to study the subcellular localisation of diacylglycerol (DAG), a modulator of membrane morphology and membrane dynamics in nuclear envelope assembly. We show that DAG is localised to the nuclear envelope, nucleoplasmic reticulum and curved tips of the Golgi apparatus. With these developments, we demonstrate that integrated imaging is maturing into a powerful tool for accurate molecular localisation to structure. PMID:24637200

  14. IceCube simulation production and the transition to IceProd2

    NASA Astrophysics Data System (ADS)

    Schultz, David

    2016-04-01

    IceCube's simulation production relies largely on dynamic, heterogeneous resources spread around the world. Datasets consist of many thousands of job workflow subsets running in parallel as directed acyclic graphs (DAGs) and using varying resources. IceProd is a set of Python daemons which process job workflow and maintain configuration and status information on jobs before, during, and after processing. IceProd manages a complex workflow of DAGs to distribute jobs across all computing grids and optimize resource usage. IceProd2 is a new version of IceProd with substantial increases in security, reliability, scalability, and ease of use. It is undergoing testing and will be deployed this fall.

  15. Correlative and integrated light and electron microscopy of in-resin GFP fluorescence, used to localise diacylglycerol in mammalian cells

    PubMed Central

    Peddie, Christopher J.; Blight, Ken; Wilson, Emma; Melia, Charlotte; Marrison, Jo; Carzaniga, Raffaella; Domart, Marie-Charlotte; O׳Toole, Peter; Larijani, Banafshe; Collinson, Lucy M.

    2014-01-01

    Fluorescence microscopy of GFP-tagged proteins is a fundamental tool in cell biology, but without seeing the structure of the surrounding cellular space, functional information can be lost. Here we present a protocol that preserves GFP and mCherry fluorescence in mammalian cells embedded in resin with electron contrast to reveal cellular ultrastructure. Ultrathin in-resin fluorescence (IRF) sections were imaged simultaneously for fluorescence and electron signals in an integrated light and scanning electron microscope. We show, for the first time, that GFP is stable and active in resin sections in vacuo. We applied our protocol to study the subcellular localisation of diacylglycerol (DAG), a modulator of membrane morphology and membrane dynamics in nuclear envelope assembly. We show that DAG is localised to the nuclear envelope, nucleoplasmic reticulum and curved tips of the Golgi apparatus. With these developments, we demonstrate that integrated imaging is maturing into a powerful tool for accurate molecular localisation to structure. PMID:24637200

  16. Diacylglycerol Kinases: Regulated Controllers of T Cell Activation, Function, and Development

    PubMed Central

    Joshi, Rohan P.; Koretzky, Gary A.

    2013-01-01

    Diacylglycerol kinases (DGKs) are a diverse family of enzymes that catalyze the conversion of diacylglycerol (DAG), a crucial second messenger of receptor-mediated signaling, to phosphatidic acid (PA). Both DAG and PA are bioactive molecules that regulate a wide set of intracellular signaling proteins involved in innate and adaptive immunity. Clear evidence points to a critical role for DGKs in modulating T cell activation, function, and development. More recently, studies have elucidated factors that control DGK function, suggesting an added complexity to how DGKs act during signaling. This review summarizes the available knowledge of the function and regulation of DGK isoforms in signal transduction with a particular focus on T lymphocytes. PMID:23531532

  17. Advanced Air Transportation Technologies (AATT) Project: Distributed Air-Ground Traffic Management

    NASA Technical Reports Server (NTRS)

    Mogford, Richard; Green, Steve; Ballin, Mark

    2002-01-01

    This viewgraph presentation provides an overview of active Distributed Air Ground Traffic Management (DAG-TM) work and reported on its overall progress to date. It does not include details on the concept elements (CEs).The DAG-TM research project is defined as a concept development and definition project and no tools will be delivered. Of the 14 CEs, three are being explored actively: CE-5, CE-6, and CE-11. Overviews of CE-5 (Free Maneuvering for User-Preferred Separation Assurance and Local TFM Conformance), CE-6 (En Route and Transition Trajectory Negotiation for User-Preferred Separation and Local TFM Conformance) and CE-11 (Self-Spacing for Merging and In-Trail Separation) are presented.

  18. Aluminum ions alter the function of non-specific phospholipase C through the changes in plasma membrane physical properties.

    PubMed

    Pejchar, Přemysl; Martinec, Jan

    2015-01-01

    The first indication of the aluminum (Al) toxicity in plants growing in acidic soils is the cessation of root growth, but the detailed mechanism of Al effect is unknown. Here we examined the impact of Al stress on the activity of non-specific phospholipase C (NPC) in the connection with the processes related to the plasma membrane using fluorescently labeled phosphatidylcholine. We observed a rapid and significant decrease of labeled diacylglycerol (DAG), product of NPC activity, in Arabidopsis seedlings treated with AlCl₃. Interestingly, an application of the membrane fluidizer, benzyl alcohol, restored the level of DAG during Al treatment. Our observations suggest that the activity of NPC is affected by Al-induced changes in plasma membrane physical properties. PMID:26024014

  19. Aluminum ions alter the function of non-specific phospholipase C through the changes in plasma membrane physical properties

    PubMed Central

    Pejchar, Přemysl; Martinec, Jan

    2015-01-01

    The first indication of the aluminum (Al) toxicity in plants growing in acidic soils is the cessation of root growth, but the detailed mechanism of Al effect is unknown. Here we examined the impact of Al stress on the activity of non-specific phospholipase C (NPC) in the connection with the processes related to the plasma membrane using fluorescently labeled phosphatidylcholine. We observed a rapid and significant decrease of labeled diacylglycerol (DAG), product of NPC activity, in Arabidopsis seedlings treated with AlCl3. Interestingly, an application of the membrane fluidizer, benzyl alcohol, restored the level of DAG during Al treatment. Our observations suggest that the activity of NPC is affected by Al-induced changes in plasma membrane physical properties. PMID:26024014

  20. A chloroplast pathway for the de novo biosynthesis of triacylglycerol in Chlamydomonas reinhardtii

    SciTech Connect

    Fan, J.; Xu, C.; Andre, C.

    2011-06-23

    Neutral lipid metabolism has been extensively studied in yeast, plants and mammals. In contrast, little information is available regarding the biochemical pathway, enzymes and regulatory factors involved in the biosynthesis of triacylglycerol (TAG) in microalgae. In the conventional TAG biosynthetic pathway widely accepted for yeast, plants and mammals, TAG is assembled in the endoplasmic reticulum (ER) from its immediate precursor diacylglycerol (DAG) made by ER-specific acyltransferases, and is deposited exclusively in lipid droplets in the cytosol. Here, we demonstrated that the unicellular microalga Chlamydomonas reinhardtii employs a distinct pathway that uses DAG derived almost exclusively from the chloroplast to produce TAG. This unique TAG biosynthesis pathway is largely dependent on de novo fatty acid synthesis, and the TAG formed in this pathway is stored in lipid droplets in both the chloroplast and the cytosol. These findings have wide implications for understanding TAG biosynthesis and storage and other areas of lipid metabolism in microalgae and other organisms.

  1. Analyzing microarray data with transitive directed acyclic graphs.

    PubMed

    Phan, Vinhthuy; Olusegun George, E; Tran, Quynh T; Goodwin, Shirlean; Bodreddigari, Sridevi; Sutter, Thomas R

    2009-02-01

    Post hoc assignment of patterns determined by all pairwise comparisons in microarray experiments with multiple treatments has been proven to be useful in assessing treatment effects. We propose the usage of transitive directed acyclic graphs (tDAG) as the representation of these patterns and show that such representation can be useful in clustering treatment effects, annotating existing clustering methods, and analyzing sample sizes. Advantages of this approach include: (1) unique and descriptive meaning of each cluster in terms of how genes respond to all pairs of treatments; (2) insensitivity of the observed patterns to the number of genes analyzed; and (3) a combinatorial perspective to address the sample size problem by observing the rate of contractible tDAG as the number of replicates increases. The advantages and overall utility of the method in elaborating drug structure activity relationships are exemplified in a controlled study with real and simulated data. PMID:19226664

  2. ADS-B within a Multi-Aircraft Simulation for Distributed Air-Ground Traffic Management

    NASA Technical Reports Server (NTRS)

    Barhydt, Richard; Palmer, Michael T.; Chung, William W.; Loveness, Ghyrn W.

    2004-01-01

    Automatic Dependent Surveillance Broadcast (ADS-B) is an enabling technology for NASA s Distributed Air-Ground Traffic Management (DAG-TM) concept. DAG-TM has the goal of significantly increasing capacity within the National Airspace System, while maintaining or improving safety. Under DAG-TM, aircraft exchange state and intent information over ADS-B with other aircraft and ground stations. This information supports various surveillance functions including conflict detection and resolution, scheduling, and conformance monitoring. To conduct more rigorous concept feasibility studies, NASA Langley Research Center s PC-based Air Traffic Operations Simulation models a 1090 MHz ADS-B communication structure, based on industry standards for message content, range, and reception probability. The current ADS-B model reflects a mature operating environment and message interference effects are limited to Mode S transponder replies and ADS-B squitters. This model was recently evaluated in a Joint DAG-TM Air/Ground Coordination Experiment with NASA Ames Research Center. Message probability of reception vs. range was lower at higher traffic levels. The highest message collision probability occurred near the meter fix serving as the confluence for two arrival streams. Even the highest traffic level encountered in the experiment was significantly less than the industry standard "LA Basin 2020" scenario. Future studies will account for Mode A and C message interference (a major effect in several industry studies) and will include Mode A and C aircraft in the simulation, thereby increasing the total traffic level. These changes will support ongoing enhancements to separation assurance functions that focus on accommodating longer ADS-B information update intervals.

  3. Activation of protein kinase C by lysophosphatidic acid: dependence on composition of phospholipid vesicles.

    PubMed Central

    Sando, J J; Chertihin, O I

    1996-01-01

    Lysophosphatidic acid (LPA) has attracted recent attention as a major serum-derived regulator implicated in responses to vascular injury and inflammation, in tumour invasiveness and in neuronal signalling and remodelling. Although the possibility of a specific G-protein-coupled LPA receptor protein has been suggested, characterization of such a receptor is lacking. Since LPA can activate protein kinase C (PKC) pathways in many cells and PKC activators mimic many LPA effects, the possibility of more direct LPA effects on PKC was investigated. Phosphatidylcholine (PC)/phosphatidylserine (PS)/diacylglycerol (DAG) lipid vesicles of defined acyl chain composition were used to activate the enzyme. At total concentrations of saturated PC/PS + DAG vesicles (2-3 mM) that provided maximal PKC activation, 1-10 mol % [18:1]-LPA led to a further approx. 2-fold activation of PKC alpha. At lower lipid concentrations, a greater increase was observed with LPA concentrations up to 16-20 mol %. Higher concentrations of LPA were inhibitory. The LPA activation of PKC was dependent on the presence of DAG, PS and Ca2+. [18:1]-Lysophosphatidylcholine produced similar PKC activation in PC/PS/DAG vesicles. [14:0]-LPA was less effective, and longer-chain saturated lysolipids were ineffective. In unsaturated PC/PS vesicles, very little to no effect of LPA was discernable. These results suggest that physiologically or pathologically relevant concentrations of LPA can contribute to PKC activation depending on the composition of the lipid membrane. We hypothesize that LPA may affect the formation of lipid domains that are recognized by the enzyme. PMID:8713089

  4. TRP_2, a lipid/trafficking domain that mediates diacylglycerol-induced vesicle fusion.

    PubMed

    van Rossum, Damian B; Oberdick, Daniel; Rbaibi, Youssef; Bhardwaj, Gaurav; Barrow, Roxanne K; Nikolaidis, Nikolas; Snyder, Solomon H; Kiselyov, Kirill; Patterson, Randen L

    2008-12-01

    We recently modeled transient receptor potential (TRP) channels using the Gestalt Domain Detection Algorithm-Basic Local Alignment Tool (GDDA-BLAST), which derives structural, functional, and evolutionary information from primary amino acid sequences using phylogenetic profiles ( Ko, K. D., Hong, Y., Chang, G. S., Bhardwaj, G., van Rossum, D. B., and Patterson, R. L. (2008) Physics Arch. Quant. Methods arXiv: 0806.2394v1 ). Herein we test our functional predictions for the TRP_2 domain of TRPC3; a domain of unknown function that is conserved in all TRPC channels. Our functional models of this domain identify both lipid binding and trafficking activities. In this study, we reveal: (i) a novel structural determinant of ion channel sensitivity to lipids, (ii) a molecular mechanism for the difference between diacylglycerol (DAG)-sensitive and DAG-insensitive TRPC subfamilies, and (iii) evidence that TRPC3 can comprise part of the vesicle fusion machinery. Indeed, the TRPC3 TRP_2 domain mediates channel trafficking to the plasma membrane and binds to plasma membrane lipids. Further, mutations in TRP_2, which alter lipid binding, also disrupt the DAG-mediated fusion of TRPC3-containing vesicles with the plasma membrane without disrupting SNARE interactions. Importantly, these data agree with the known role of DAG in membrane destabilization, which facilitates SNARE-dependent synaptic vesicle fusion ( Villar, A. V., Goni, F. M., and Alonso, A. (2001) FEBS Lett. 494, 117-120 and Goni, F. M., and Alonso, A. (1999) Prog. Lipid Res. 38, 1-48 ). Taken together, functional models generated by GDDA-BLAST provide a computational platform for deriving domain functionality, which can have in vivo and mechanistic relevance. PMID:19043047

  5. The occurrence of domoic acid linked to a toxic diatom bloom in a new potential vector: the tunicate Pyura chilensis (piure).

    PubMed

    López-Rivera, Américo; Pinto, Maricela; Insinilla, Andrea; Suárez Isla, Benjamín; Uribe, Eduardo; Alvarez, Gonzalo; Lehane, Mary; Furey, Ambrose; James, Kevin J

    2009-11-01

    The tunicate Pyura chilensis (Molina, 1782); Phylum Chordata; Subphylum Urochordata; Class Ascidiacea, common local name "piure" or sea squirt; a filter-feeder (plankton and suspended particles) sessile species; may play an important role in monitoring domoic acid (DA) the principal toxic component of Amnesic Shellfish Poisoning (ASP). Significant DA concentrations have been determined in tunicate samples, collected during a recent ASP outbreak in Bahía Inglesa, an important scallop (Argopecten purpuratus) farming area. Several infaunal species were tested for the presence of DA, in addition to the usual scallop monitoring programme. DA was found at sub-toxic levels in filtering bivalves such as mussels (Mytilus chilensis), large mussels (Aulacomya ater) and clams (Protothaca thaca) (6.4, 5.4 and 4.7 microg DA/g tissue respectively). Of particular interest was the observation of significant accumulations of toxic Pseudo-nitzschia sp. diatoms in the internal siphon and atrium spaces of the tunicate. Toxin distribution within major tunicate organs was heterogeneous with 8.7-15.5 microg DA/g in edible tissues, 14.9-17.9 microg DA/g in the fecal material and 13.6-32.7 microg DA/g in the gut content. DA was determined by HPLC-UV and confirmed by diode-array detection and LC-MS/MS analysis. This is the first report of the presence of DA in a tunicate that is regularly consumed by coastal populations. These results confirm the need to include these organisms in sanitation programs for marine toxins. PMID:19505493

  6. Compositions and taxonomy of 15 unusual carbonaceous chondrites

    NASA Astrophysics Data System (ADS)

    Choe, Won Hie; Huber, Heinz; Rubin, Alan E.; Kallemeyn, Gregory W.; Wasson, John T.

    2010-04-01

    We used instrumental neutron activation analysis and petrography to determine bulk and phase compositions and textural characteristics of 15 carbonaceous chondrites of uncertain classification: Acfer 094 (type 3.0, ungrouped CM-related); Belgica-7904 (mildly metamorphosed, anomalous, CM-like chondrite, possibly a member of a new grouplet that includes Wisconsin Range (WIS) 91600, Dhofar 225, and Yamato-86720); Dar al Gani (DaG) 055 and its paired specimen DaG 056 (anomalous, reduced CV3-like); DaG 978 (type 3 ungrouped); Dominion Range 03238 (anomalous, magnetite-rich CO3.1); Elephant Moraine 90043 (anomalous, magnetite-bearing CO3); Graves Nunataks 98025 (type 2 or type 3 ungrouped); Grosvenor Mountains (GRO) 95566 (anomalous CM2 with a low degree of aqueous alteration); Hammadah al Hamra (HaH) 073 (type 4 ungrouped, possibly related to the Coolidge-Loongana [C-L] 001 grouplet); Lewis Cliff (LEW) 85311 (anomalous CM2 with a low degree of aqueous alteration); Northwest Africa 1152 (anomalous CV3); Pecora Escarpment (PCA) 91008 (anomalous, metamorphosed CM); Queen Alexandra Range 99038 (type 2 ungrouped); Sahara 00182 (type 3 ungrouped, possibly related to HaH 073 and/or to C-L 001); and WIS 91600 (mildly metamorphosed, anomalous, CM-like chondrite, possibly a member of a new grouplet that includes Belgica-7904, Dhofar 225, and Y-86720). Many of these meteorites show fractionated abundance patterns, especially among the volatile elements. Impact volatilization and dehydration as well as elemental transport caused by terrestrial weathering are probably responsible for most of these compositional anomalies. The metamorphosed CM chondrites comprise two distinct clusters on the basis of their Δ17O values: approximately -4‰ for PCA 91008, GRO 95566, DaG 978, and LEW 85311, and approximately 0‰ for Belgica-7904 and WIS 91600. These six meteorites must have been derived from different asteroidal regions.

  7. Signal-dependent Hydrolysis of Phosphatidylinositol 4,5-Bisphosphate without Activation of Phospholipase C

    PubMed Central

    Lev, Shaya; Katz, Ben; Tzarfaty, Vered; Minke, Baruch

    2012-01-01

    In Drosophila, a phospholipase C (PLC)-mediated signaling cascade, couples photo-excitation of rhodopsin to the opening of the transient receptor potential (TRP) and TRP-like (TRPL) channels. A lipid product of PLC, diacylglycerol (DAG), and its metabolites, polyunsaturated fatty acids (PUFAs) may function as second messengers of channel activation. However, how can one separate between the increase in putative second messengers, change in pH, and phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) depletion when exploring the TRPL gating mechanism? To answer this question we co-expressed the TRPL channels together with the muscarinic (M1) receptor, enabling the openings of TRPL channels via G-protein activation of PLC. To dissect PLC activation of TRPL into its molecular components, we used a powerful method that reduced plasma membrane-associated PI(4,5)P2 in HEK cells within seconds without activating PLC. Upon the addition of a dimerizing drug, PI(4,5)P2 was selectively hydrolyzed in the cell membrane without producing DAG, inositol trisphosphate, or calcium signals. We show that PI(4,5)P2 is not an inhibitor of TRPL channel activation. PI(4,5)P2 hydrolysis combined with either acidification or application of DAG analogs failed to activate the channels, whereas PUFA did activate the channels. Moreover, a reduction in PI(4,5)P2 levels or inhibition of DAG lipase during PLC activity suppressed the PLC-activated TRPL current. This suggests that PI(4,5)P2 is a crucial substrate for PLC-mediated activation of the channels, whereas PUFA may function as the channel activator. Together, this study defines a narrow range of possible mechanisms for TRPL gating. PMID:22065576

  8. Simultaneous detection of phosphatidylcholines and glycerolipids using matrix-enhanced surface-assisted laser desorption/ionization-mass spectrometry with sputter-deposited platinum film.

    PubMed

    Ozawa, Tomoyuki; Osaka, Issey; Ihozaki, Taisuke; Hamada, Satoshi; Kuroda, Yusuke; Murakami, Tatsuya; Miyazato, Akio; Kawasaki, Hideya; Arakawa, Ryuichi

    2015-11-01

    Matrix-assisted laser desorption/ionisation (MALDI) imaging mass spectrometry (IMS) allows for the simultaneous detection and imaging of several molecules in brain tissue. However, the detection of glycerolipids such as diacylglycerol (DAG) and triacylglycerol (TAG) in brain tissues is hindered in MALDI-IMS because of the ion suppression effect from excessive ion yields of phosphatidylcholine (PC). In this study, we describe an approach that employs a homogeneously deposited metal nanoparticle layer (or film) for the detection of glycerolipids in rat brain tissue sections using IMS. Surface-assisted laser desorption/ionisation IMS with sputter-deposited Pt film (Pt-SALDI-IMS) for lipid analysis was performed as a solvent-free and organic matrix-free method. Pt-SALDI produced a homogenous layer of nanoparticles over the surface of the rat brain tissue section. Highly selective detection of lipids was possible by MALDI-IMS and Pt-SALDI-IMS; MALDI-IMS detected the dominant ion peak of PC in the tissue section, and there were no ion peaks representing glycerolipids such as DAG and TAG. In contrast, Pt-SALDI-IMS allowed the detection of these glycerolipids, but not PC. Therefore, using a hybrid method combining MALDI and Pt-SALDI (i.e., matrix-enhanced [ME]-Pt-SALDI-IMS), we achieved the simultaneous detection of PC, PE and DAG in rat brain tissue sections, and the sensitivity for the detection of these molecules was better than that of MALDI-IMS or Pt-SALDI alone. The present simple ME-Pt-SALDI approach for the simultaneous detection of PC and DAG using two matrices (sputter-deposited Pt film and DHB matrix) would be useful in imaging analyses of biological tissue sections. PMID:26505771

  9. Data management and processing plan, Department of Applied Geodesy

    SciTech Connect

    Not Available

    1992-08-01

    This plan outlines Data Management and Data Processing requirements of the Department of Applied Geodesy (DAG) and presents the plan to meet these requirements (These requirements are derived from the functional needs of the Department to meet the SSCL alignment tolerances and schedules). In addition, this document presents a schedule for the implementation of this plan. This document is an integral part of the Alignment Plan of the SSCL.

  10. Assessment of intracellular Ca2+, cAMP and 1,2-diacylglycerol in cryopreserved buffalo (Bubalus bubalis) spermatozoa on supplementation of taurine and trehalose in the extender.

    PubMed

    Singh, V K; Atreja, S K; Kumar, R; Chhillar, S; Singh, A K

    2012-08-01

    In mammalian spermatozoa, intracellular calcium plays a major role in sperm functions like motility and capacitation. Cryopreservation-induced modifications to sperm membrane result in an influx of intracellular calcium affecting calcium-dependent intracellular signalling pathways. Intracellular calcium activates adenyl cyclase to produce cAMP that activates phospholipase A(2) (PLA(2) ) and phospholipase C (PLC) generating lysophosphatidyl choline, 1,2-diacylglycerol (DAG) and IP(3) , acting as intracellular secondary messengers required for sperm capacitation. Present study was designed to determine levels of intracellular calcium, cAMP and DAG in fresh and frozen-thawed buffalo spermatozoa cryopreserved in the presence and absence of taurine or trehalose. A total number of nine ejaculates from three randomly chosen buffalo bulls were cryopreserved in Tris-based egg yolk extender and thawed in warm water at 37°C. The cAMP was measured by enzyme immuno assay, and intracellular calcium was quantified using fluorescent dye FURA 2-AM. Total lipid was extracted from spermatozoa, and DAG was estimated using thin layer chromatography followed by spectrophotometric analysis. Intracellular calcium, cAMP and DAG levels in spermatozoa were significantly (p < 0.01) increased following cryopreservation as compared to fresh ejaculate. Addition of taurine or trehalose to the freezing medium significantly decreased (p < 0.01) the levels of intracellular calcium and cAMP in frozen-thawed spermatozoa. 1,2-diacylglycerol content was also decreased significantly (p < 0.01) in spermatozoa cryopreserved in presence of additives. Moreover, significant (p < 0.01) improvement in post-thaw motility, viability and membrane integrity of spermatozoa on addition of taurine or trehalose clearly indicated the reduced level of capacitation-like changes in buffalo spermatozoa. PMID:21988572

  11. Skeletal muscle fatty acid handling in insulin resistant men.

    PubMed

    van Hees, Anneke M J; Jans, Anneke; Hul, Gabby B; Roche, Helen M; Saris, Wim H M; Blaak, Ellen E

    2011-07-01

    Disturbances in skeletal muscle lipid metabolism may precede or contribute to the development of whole body insulin resistance. In this study, we examined fasting and postprandial skeletal muscle fatty acid (FA) handling in insulin resistant (IR) men. Thirty men with the metabolic syndrome (MetS) (National Cholesterol Education Program-Adult Treatment Panel III) were included in this sub-study to the LIPGENE study, and divided in two groups (IR and control) based on the median of insulin sensitivity (S(I) = 2.06 (mU/l(-1))·min(-1)·10(-4)). Fasting and postprandial skeletal muscle FA handling were examined by combining the forearm balance technique with stable isotopes of palmitate. [(2)H(2)]-palmitate was infused intravenously to label endogenous triacylglycerol (TAG) and free FAs (FFAs) in the circulation and [U-(13)C]-palmitate was incorporated in a high-fat mixed meal (2.6 MJ, 61 E% fat) to label chylomicron-TAG. Muscle biopsies were taken to determine muscle TAG, diacylglycerol (DAG), FFA, and phospholipid (PL) content, their fractional synthetic rates (FSRs) and degree of saturation, as well as messenger RNA (mRNA) expression of genes involved in lipid metabolism. In the first 2 h after meal consumption, forearm muscle [(2)H(2)]-labeled TAG extraction was higher in IR vs. control (P = 0.05). Fasting percentage saturation of muscle DAG was higher in IR vs. control (P = 0.016). No differences were observed for intramuscular TAG, DAG, FFA, and PL content, FSR, and muscle mRNA expression. In conclusion, increased muscle (hepatically derived) TAG extraction during postprandial conditions and increased saturation of intramuscular DAG are associated with insulin resistance, suggesting that disturbances in skeletal muscle FA handling could play a role in the development of whole body insulin resistance and type 2 diabetes. PMID:21331063

  12. TRP_2, a Lipid/Trafficking Domain That Mediates Diacylglycerol-induced Vesicle Fusion*S⃞

    PubMed Central

    van Rossum, Damian B.; Oberdick, Daniel; Rbaibi, Youssef; Bhardwaj, Gaurav; Barrow, Roxanne K.; Nikolaidis, Nikolas; Snyder, Solomon H.; Kiselyov, Kirill; Patterson, Randen L.

    2008-01-01

    We recently modeled transient receptor potential (TRP) channels using the Gestalt Domain Detection Algorithm-Basic Local Alignment Tool (GDDA-BLAST), which derives structural, functional, and evolutionary information from primary amino acid sequences using phylogenetic profiles (Ko, K. D., Hong, Y., Chang, G. S., Bhardwaj, G., van Rossum, D. B., and Patterson, R. L. (2008) Physics Arch. Quant. Methods arXiv:0806.2394v1). Herein we test our functional predictions for the TRP_2 domain of TRPC3; a domain of unknown function that is conserved in all TRPC channels. Our functional models of this domain identify both lipid binding and trafficking activities. In this study, we reveal: (i) a novel structural determinant of ion channel sensitivity to lipids, (ii) a molecular mechanism for the difference between diacylglycerol (DAG)-sensitive and DAG-insensitive TRPC subfamilies, and (iii) evidence that TRPC3 can comprise part of the vesicle fusion machinery. Indeed, the TRPC3 TRP_2 domain mediates channel trafficking to the plasma membrane and binds to plasma membrane lipids. Further, mutations in TRP_2, which alter lipid binding, also disrupt the DAG-mediated fusion of TRPC3-containing vesicles with the plasma membrane without disrupting SNARE interactions. Importantly, these data agree with the known role of DAG in membrane destabilization, which facilitates SNARE-dependent synaptic vesicle fusion (Villar, A. V., Goni, F. M., and Alonso, A. (2001) FEBS Lett. 494,117 -12011297746 and Goni, F. M., and Alonso, A. (1999) Prog. Lipid Res. 38,1 -4810396601). Taken together, functional models generated by GDDA-BLAST provide a computational platform for deriving domain functionality, which can have in vivo and mechanistic relevance. PMID:19043047

  13. Expression of Mouse MGAT in Arabidopsis Results in Increased Lipid Accumulation in Seeds

    PubMed Central

    El Tahchy, Anna; Petrie, James R.; Shrestha, Pushkar; Vanhercke, Thomas; Singh, Surinder P.

    2015-01-01

    Worldwide demand for vegetable oil is projected to double within the next 30 years due to increasing food, fuel, and industrial requirements. There is therefore great interest in metabolic engineering strategies that boost oil accumulation in plant tissues, however, efforts to date have only achieved levels of storage lipid accumulation in plant tissues far below the benchmark to meet demand. Monoacylglycerol acyltransferase (MGAT) is predominantly associated with lipid absorption and resynthesis in the animal intestine where it catalyzes monoacylglycerol (MAG) to form diacylglycerol (DAG), and then triacylglycerol (TAG). In contrast plant lipid biosynthesis routes do not include MGAT. Rather, DAG and TAG are either synthesized from glycerol-3-phosphate by a series of three subsequent acylation reactions, or originated from phospholipids via an acyl editing pathway. Mouse MGATs 1 and 2 have been shown to increase oil content transiently in Nicotiana benthamiana leaf tissue by 2.6 fold. Here we explore the feasibility of this approach to increase TAG in Arabidopsis thaliana seed. The stable MGAT2 expression resulted in a significant increase in seed oil content by 1.32 fold. We also report evidence of the MGAT2 activity based on in vitro assays. Up to 3.9 fold increase of radiolabeled DAG were produced in seed lysate which suggest that the transgenic MGAT activity can result in DAG re-synthesis by salvaging the MAG product of lipid breakdown. The expression of MGAT2 therefore creates an independent and complementary TAG biosynthesis route to the endogenous Kennedy pathway and other glycerolipid synthesis routes. PMID:26834753

  14. Three essays on price dynamics and causations among energy markets and macroeconomic information

    NASA Astrophysics Data System (ADS)

    Hong, Sung Wook

    This dissertation examines three important issues in energy markets: price dynamics, information flow, and structural change. We discuss each issue in detail, building empirical time series models, analyzing the results, and interpreting the findings. First, we examine the contemporaneous interdependencies and information flows among crude oil, natural gas, and electricity prices in the United States (US) through the multivariate generalized autoregressive conditional heteroscedasticity (MGARCH) model, Directed Acyclic Graph (DAG) for contemporaneous causal structures and Bernanke factorization for price dynamic processes. Test results show that the DAG from residuals of out-of-sample-forecast is consistent with the DAG from residuals of within-sample-fit. The result supports innovation accounting analysis based on DAGs using residuals of out-of-sample-forecast. Second, we look at the effects of the federal fund rate and/or WTI crude oil price shock on US macroeconomic and financial indicators by using a Factor Augmented Vector Autoregression (FAVAR) model and a graphical model without any deductive assumption. The results show that, in contemporaneous time, the federal fund rate shock is exogenous as the identifying assumption in the Vector Autoregression (VAR) framework of the monetary shock transmission mechanism, whereas the WTI crude oil price return is not exogenous. Third, we examine price dynamics and contemporaneous causality among the price returns of WTI crude oil, gasoline, corn, and the S&P 500. We look for structural break points and then build an econometric model to find the consistent sub-periods having stable parameters in a given VAR framework and to explain recent movements and interdependency among returns. We found strong evidence of two structural breaks and contemporaneous causal relationships among the residuals, but also significant differences between contemporaneous causal structures for each sub-period.

  15. Diacylglycerol-containing docosahexaenoic acid in acyl chain modulates airway smooth muscle tone.

    PubMed

    Hichami, Aziz; Morin, Caroline; Rousseau, Eric; Khan, Naim A

    2005-10-01

    We synthesized and assessed the role of a diacylglycerol (DAG)-containing docosahexaenoic acid (DHA), that is, 1-stearoyl-2-docosahexaenoyl-sn-glycerol (SDHG), in the contraction of guinea pig airway smooth muscle (ASM). We compared its action with 1-stearoyl-2-arachidonoyl-sn-glycerol (SAG) and 1,2-dioctanoyl-sn-glycerol (1,2-DiC8), a stable DAG analog. The three DAGs (SAG, SDHG, and 1,2-DiC8) induced reversible concentration-dependent contraction of ASM. SDHG induced higher guinea pig ASM contraction than did SAG and 1,2-DiC8. The effects of SDHG were blocked, to different extents, by nifedipine (L-type Ca2+ channel blocker). By employing GF-109203X (protein kinase C [PKC] inhibitor) and lanthanum (La3+), a nonselective cation channel blocker, we observed that SDHG evoked ASM contractile response via PKC-dependent and PKC-independent (but Ca2+-dependent) pathways. Interestingly, SAG exerted its action only by increasing [Ca2+]i and did not require PKC activation. To probe the implication of calcium mobilization, we employed thapsigargin (TG), which also induced ASM contraction in a calcium-dependent manner. SDHG and 1,2-DiC8, in a PKC-dependent manner, induced the phosphorylation of CPI-17 (myosin light chain phosphatase inhibitor of 17 kD). Furthermore, SAG and TG failed to phosphorylate CPI-17 in ASM cells. Our results suggest that different DAG species, produced during a dietary supplementation with fatty acids, could modulate the reactivity of airway smooth muscles in a PKC-dependent and -independent manner, and hence, may play a critical role in health and disease. PMID:15961724

  16. TRP channels and lipids: from Drosophila to mammalian physiology

    PubMed Central

    Hardie, Roger C

    2007-01-01

    The transient receptor potential (TRP) ion channel family was the last major ion channel family to be discovered. The prototypical member (dTRP) was identified by a forward genetic approach in Drosophila, where it represents the transduction channel in the photoreceptors, activated downstream of a Gq-coupled PLC. In the meantime 29 vertebrate TRP isoforms are recognized, distributed amongst seven subfamilies (TRPC, TRPV, TRPM, TRPML, TRPP, TRPA, TRPN). They subserve a wide range of functions throughout the body, most notably, though by no means exclusively, in sensory transduction and in vascular smooth muscle. However, their precise physiological roles and mechanism of activation and regulation are still only gradually being revealed. Most TRP channels are subject to multiple modes of regulation, but a common theme amongst the TRPC/V/M subfamilies is their regulation by lipid messengers. Genetic evidence supports an excitatory role of diacylglycerol (DAG) for the dTRP's, although curiously only DAG metabolites (PUFAs) have been found to activate the Drosophila channels. TRPC2,3,6 and 7 are widely accepted as DAG-activated channels, although TRPC3 can also be regulated via a store-operated mechanism. More recently PIP2 has been shown to be required for activity of TRPV5, TRPM4,5,7 and 8, whilst it may inhibit TRPV1 and the dTRPs. Although compelling evidence for a direct interaction of DAG with the TRPC channels is lacking, mutagenesis studies have identified putative PIP2-interacting domains in the C-termini of several TRPV and TRPM channels. PMID:16990401

  17. Signaling Responses to Pulsatile Gonadotropin-releasing Hormone in LβT2 Gonadotrope Cells*

    PubMed Central

    Tsutsumi, Rie; Mistry, Devendra; Webster, Nicholas J. G.

    2010-01-01

    The hypothalamic neuropeptide gonadotropin-releasing hormone (GnRH) is secreted in a pulsatile fashion by hypothalamic neurons, and alterations in pulse frequency and amplitude differentially regulate gonadotropin synthesis and release. In this study, we investigated the kinetics of Gs and Gq signaling in response to continuous or pulsatile GnRH using fluorescence resonance energy transfer reporters in live mouse LβT2 gonadotrope cells. cAMP and protein kinase A-dependent reporters showed a rapid but transient increase in fluorescence resonance energy transfer signal with increasing doses of constant GnRH, and in contrast diacylglycerol (DAG) and calcium reporters showed a rapid and sustained signal. Multiple pulses of GnRH caused multiple pulses of cAMP and protein kinase A activation without desensitization, but the DAG and calcium reporters were rapidly desensitized resulting in inhibition of calcium and DAG responses. At the transcriptional level, both a cAMP-dependent cAMP-response element reporter and a DAG/calcium-dependent AP-1 reporter showed a pulse frequency-dependent increase in luciferase activity. However, constant GnRH stimulation gave very little cAMP-response element activation but very strong AP-1 activation. Based on these data, we propose that both the GnRH-R-Gs and Gq pathways are responsive to pulses of GnRH, but only the Gq pathway is responsive to constant GnRH. Furthermore, the Gq pathway is subject to desensitization with multiple GnRH pulses, but the Gs pathway is not. PMID:20406815

  18. Rising Nationalism vs. The International Language of Advertising.

    ERIC Educational Resources Information Center

    Dunn, S. Watson

    One of the cherished myths of the 1960s and the 1970s has been that the countries of the world are at last giving up their long-standing nationalism and are approaching the one world that Wendell Wilkie, Dag Hammarskjold, and many others used to talk about. There is indeed much evidence to indicate that this may be true. On the other hand, Peter…

  19. Differential effects of saturated versus unsaturated dietary fatty acids on weight gain and myocellular lipid profiles in mice

    PubMed Central

    Timmers, S; de Vogel-van den Bosch, J; de Wit, N; Schaart, G; van Beurden, D; Hesselink, M; van der Meer, R; Schrauwen, P

    2011-01-01

    Objective: In conditions of continuous high-fat (HF) intake, the degree of saturation of the fatty acids (FAs) in the diet might have a crucial role in the onset of obesity and its metabolic complications. In particular, the FA composition of the diet might influence the storage form of lipids inside skeletal muscle. The aim of the present study was to examine whether the FA composition of HF diets differentially affects weight gain and accumulation of myocellular triacylglycerol (TAG) and diacylglycerol (DAG). Furthermore, we examined whether the FA composition of the diet was reflected in the composition of the myocellular lipid intermediates. Design: C57Bl6 mice were fed HF diets (45% energy) mainly containing palm oil (PO), cocoa butter (CB), olive oil (OO) or safflower oil (SO; n=6 per group) for 8 weeks. A low-fat diet (10% energy, PO) was used as control. Body weight was monitored weekly. At the end of the dietary intervention, myocellular TAG and DAG content and profiles were measured. Results: We here show that HF_CB prevented weight gain after 8 weeks of HF feeding. Furthermore, the HF diet rich in SO prevented the accumulation of both myocellular TAG and DAG. Interestingly, the FA composition of DAG and TAG in skeletal muscle was a reflection of the dietary FA composition. Conclusion: Already after a relatively short period, the dietary FA intake relates to the FA composition of the lipid metabolites in the muscle. A diet rich in polyunsaturated FAs seems to prevent myocellular lipid accumulation. PMID:23449423

  20. Differential signaling during macropinocytosis in response to M-CSF and PMA in macrophages

    PubMed Central

    Yoshida, Sei; Gaeta, Isabella; Pacitto, Regina; Krienke, Lydia; Alge, Olivia; Gregorka, Brian; Swanson, Joel A.

    2015-01-01

    The cellular movements that construct a macropinosome have a corresponding sequence of chemical transitions in the cup-shaped region of plasma membrane that becomes the macropinosome. To determine the relative positions of type I phosphatidylinositol 3-kinase (PI3K) and phospholipase C (PLC) in this pathway, we analyzed macropinocytosis in macrophages stimulated by the growth factor macrophage-colony-stimulating factor (M-CSF) and by the diacylglycerol (DAG) analog phorbol 12-myristate 13-acetate (PMA). In cells stimulated with M-CSF, microscopic imaging of fluorescent probes for intracellular lipids indicated that the PI3K product phosphatidylinositol (3,4,5)-trisphosphate (PIP3) appeared in cups just prior to DAG. We then tested the hypothesis that PMA and DAG function after PI3K and prior to Ras and protein kinase C (PKC) during macropinosome formation in macrophages. Although the PI3K target Akt was activated by M-CSF, the Akt inhibitor MK-2206 did not inhibit macropinocytosis. The phospholipase C (PLC) inhibitor U73122 blocked macropinocytosis by M-CSF but not PMA. Macropinocytosis in response to M-CSF and PMA was inhibited by the Ras inhibitor farnesyl thiosalicylate (FTS), by the PKC inhibitor Calphostin C and by the broad specificity inhibitor rottlerin. These studies support a model in which M-CSF stimulates PI3K in macropinocytic cups, and the resulting increase in PIP3 activates PLC, which in turn generates DAG necessary for activation of PKC, Ras and the late stages of macropinosome closure. PMID:25688212

  1. Screening of Transient Receptor Potential Canonical Channel Activators Identifies Novel Neurotrophic Piperazine Compounds.

    PubMed

    Sawamura, Seishiro; Hatano, Masahiko; Takada, Yoshinori; Hino, Kyosuke; Kawamura, Tetsuya; Tanikawa, Jun; Nakagawa, Hiroshi; Hase, Hideharu; Nakao, Akito; Hirano, Mitsuru; Rotrattanadumrong, Rachapun; Kiyonaka, Shigeki; Mori, Masayuki X; Nishida, Motohiro; Hu, Yaopeng; Inoue, Ryuji; Nagata, Ryu; Mori, Yasuo

    2016-03-01

    Transient receptor potential canonical (TRPC) proteins form Ca(2+)-permeable cation channels activated upon stimulation of metabotropic receptors coupled to phospholipase C. Among the TRPC subfamily, TRPC3 and TRPC6 channels activated directly by diacylglycerol (DAG) play important roles in brain-derived neurotrophic factor (BDNF) signaling, promoting neuronal development and survival. In various disease models, BDNF restores neurologic deficits, but its therapeutic potential is limited by its poor pharmacokinetic profile. Elucidation of a framework for designing small molecules, which elicit BDNF-like activity via TRPC3 and TRPC6, establishes a solid basis to overcome this limitation. We discovered, through library screening, a group of piperazine-derived compounds that activate DAG-activated TRPC3/TRPC6/TRPC7 channels. The compounds [4-(5-chloro-2-methylphenyl)piperazin-1-yl](3-fluorophenyl)methanone (PPZ1) and 2-[4-(2,3-dimethylphenyl)piperazin-1-yl]-N-(2-ethoxyphenyl)acetamide (PPZ2) activated, in a dose-dependent manner, recombinant TRPC3/TRPC6/TRPC7 channels, but not other TRPCs, in human embryonic kidney cells. PPZ2 activated native TRPC6-like channels in smooth muscle cells isolated from rabbit portal vein. Also, PPZ2 evoked cation currents and Ca(2+) influx in rat cultured central neurons. Strikingly, both compounds induced BDNF-like neurite growth and neuroprotection, which were abolished by a knockdown or inhibition of TRPC3/TRPC6/TRPC7 in cultured neurons. Inhibitors of Ca(2+) signaling pathways, except calcineurin, impaired neurite outgrowth promotion induced by PPZ compounds. PPZ2 increased activation of the Ca(2+)-dependent transcription factor, cAMP response element-binding protein. These findings suggest that Ca(2+) signaling mediated by activation of DAG-activated TRPC channels underlies neurotrophic effects of PPZ compounds. Thus, piperazine-derived activators of DAG-activated TRPC channels provide important insights for future development of a

  2. Lipase-Catalyzed Glycerolysis of Soybean and Canola Oils in a Free Organic Solvent System Assisted by Ultrasound.

    PubMed

    Remonatto, Daniela; Santin, Claudia M Trentin; Valério, Alexsandra; Lerin, Lindomar; Batistella, Luciane; Ninow, Jorge Luiz; de Oliveira, J Vladimir; de Oliveira, Débora

    2015-06-01

    This work shows new and promising experimental data of soybean oil and canola oil glycerolysis using Novozym 435 enzyme as catalyst in a solvent-free system using ultrasound bath for the emulsifier, monoglyceride (MAG), and diacylglycerol (DAG) production. The experiments were conducted in batch mode to study the influence of process variables as temperature (40 to 70 °C), immobilized enzyme content (2.5 to 10 wt%, relative to substrates), molar ratio glycerol/oil (0.8:1 to 3:1), agitation (0 to 1200 rpm) and ultrasound intensity (0 to 132 W cm(-2)). Highest yields of DAG+MAG (75 wt%) were obtained with molar ratio glycerol/canola oil 0.8:1, 70 °C, 900 rpm, 120 min of reaction time, 10 wt% of enzyme concentration, and 52.8 W cm(-2) of ultrasound intensity. When soybean oil was used, the best results in terms of DAG+MAGs (65 wt%) were using molar ratio of glycerol/soybean oil 0.8:1, 70 °C, 900 rpm, 90 min of reaction time, 10 wt% of enzyme content, and 40 % of ultrasound intensity (52.8 W cm(-2)). The results showed that the lipase-catalyzed glycerolysis in a solvent-free system with ultrasound bath can be a potential route for high content production of DAGs and MAGs. PMID:25875788

  3. Leptin, skeletal muscle lipids, and lipid-induced insulin resistance.

    PubMed

    Dube, John J; Bhatt, Bankim A; Dedousis, Nikolas; Bonen, Arend; O'Doherty, Robert M

    2007-08-01

    Leptin-induced increases in insulin sensitivity are well established and may be related to the effects of leptin on lipid metabolism. However, the effects of leptin on the levels of lipid metabolites implicated in pathogenesis of insulin resistance and the effects of leptin on lipid-induced insulin resistance are unknown. The current study addressed in rats the effects of hyperleptinemia (HL) on insulin action and markers of skeletal muscle (SkM) lipid metabolism in the absence or presence of acute hyperlipidemia induced by an infusion of a lipid emulsion. Compared with controls (CONT), HL increased insulin sensitivity, as assessed by hyperinsulinemic-euglycemic clamp ( approximately 15%), and increased SkM Akt ( approximately 30%) and glycogen synthase kinase 3 alpha ( approximately 52%) phosphorylation. These improvements in insulin action were associated with decreased SkM triglycerides (TG; approximately 61%), elevated ceramides ( approximately 50%), and similar diacylglycerol (DAG) levels in HL compared with CONT. Acute hyperlipidemia in CONT decreased insulin sensitivity ( approximately 25%) and increased SkM DAG ( approximately 33%) and ceramide ( approximately 60%) levels. However, hyperlipidemia did not induce insulin resistance or SkM DAG and ceramide accumulation in HL. SkM total fatty acid transporter CD36, plasma membrane fatty acid binding protein, acetyl Co-A carboxylase phosphorylation, and fatty acid oxidation were similar in HL compared with CONT. However, HL decreased SkM protein kinase C theta (PKC theta), a kinase implicated in mediating the detrimental effects of lipids on insulin action. We conclude that increases in insulin sensitivity induced by HL are associated with decreased levels of SkM TG and PKC theta and increased SkM insulin signaling, but not with decreases in other lipid metabolites implicated in altering SkM insulin sensitivity (DAG and ceramide). Furthermore, insulin resistance induced by an acute lipid infusion is prevented by

  4. Conflict Resolution Performance in an Experimental Study of En Route Free Maneuvering Operations

    NASA Technical Reports Server (NTRS)

    Doble, Nathan A.; Barhydt, Richard; Hitt, James M., II

    2005-01-01

    NASA has developed a far-term air traffic management concept, termed Distributed Air/Ground Traffic Management (DAG-TM). One component of DAG-TM, En Route Free Maneuvering, allows properly trained flight crews of equipped autonomous aircraft to assume responsibility for separation from other autonomous aircraft and from Instrument Flight Rules (IFR) aircraft. Ground-based air traffic controllers continue to separate IFR traffic and issue flow management constraints to all aircraft. To examine En Route Free Maneuvering operations, a joint human-in-the-loop experiment was conducted in summer 2004 at the NASA Ames and Langley Research Centers. Test subject pilots used desktop flight simulators to resolve traffic conflicts and adhere to air traffic flow constraints issued by subject controllers. The experimental airspace integrated both autonomous and IFR aircraft at varying traffic densities. This paper presents a subset of the En Route Free Maneuvering experimental results, focusing on airborne and ground-based conflict resolution, and the effects of increased traffic levels on the ability of pilots and air traffic controllers to perform this task. The results show that, in general, increases in autonomous traffic do not significantly impact conflict resolution performance. In addition, pilot acceptability of autonomous operations remains high throughout the range of traffic densities studied. Together with previously reported findings, these results continue to support the feasibility of the En Route Free Maneuvering component of DAG-TM.

  5. The Influence of Terrestrial Weathering on Implanted Solar Gases in Lunar Meteorites

    NASA Astrophysics Data System (ADS)

    Franchi, I. A.; Verchovsky, A. V.; Pillinger, C. T.

    2000-01-01

    The recent discovery of two large lunar meteorites in the Dar al Gani (DaG) region of the Libyan Sahara desert (DaG 262 and DaG 400) have sparked considerable and wide ranging interest in lunar material. The addition of lunar meteorites to the collections of extraterrestrial material have greatly complemented the suite of samples returned by the Apollo and Luna missions almost 30 years ago, extending the range of materials available for investigation. However, care may be required in interpreting variations seen in the lunar meteorite population and also any differences with the returned lunar samples. As has been noted previously in the comparison of ordinary chondrite populations from Antarctica, hot deserts and observed falls, variations between the populations may be attributed to their interaction with the different terrestrial environments they have been exposed to rather than indigenous, pre-atmospheric differences. In the case of the lunar samples this effect may be even more extreme as all the lunar meteorites are finds whereas the returned lunar samples were collected and subsequently stored in as near to ideal conditions as practically possible. Other factors related to the high levels of radiation damage suffered by materials in the lunar regolith may also make certain aspects of the lunar meteorites susceptible to terrestrial weathering. The particular interest behind this paper is how terrestrial weathering may have influenced the abundance and release characteristics of the implanted solar gases.

  6. Metabolic pathways for the degradation of phosphatidic acid in isolated nuclei from cerebellar cells.

    PubMed

    Gaveglio, Virginia L; Pasquaré, Susana J; Giusto, Norma M

    2011-03-15

    The aim of the present research was to analyse the pathways for phosphatidic acid metabolism in purified nuclei from cerebellar cells. Lipid phosphate phosphatase and diacylglyceride lipase activities were detected in nuclei from cerebellar cells. It was observed that DAGL activity makes up 50% of LPP activity and that PtdOH can also be metabolised to lysophosphatidic acid. With a nuclear protein content of approximately 40 μg, the production of diacylglycerol and monoacylglycerol was linear for 30 min and 5 min, respectively, whereas it increased with PtdOH concentrations of up to 250 μM. LysoPtdOH, sphingosine 1-phosphate and ceramide 1-phosphate, which are alternative substrates for LPP, significantly reduced DAG production from PA. DAG and MAG production increased in the presence of Triton X-100 (1 mM) whereas no modifications were observed in the presence of ionic detergent sodium deoxycholate. Ca²+ and Mg²+ stimulated MAG production without affecting DAG formation whereas fluoride and vanadate inhibited the generation of both products. Specific PtdOH-phospholipase A1 and PtdOH-phospholipase A2 were also detected in nuclei. Our findings constitute the first reported evidence of active PtdOH metabolism involving LPP, DAGL and PtdOH-selective PLA activities in purified nuclei prepared from cerebellar cells. PMID:21216221

  7. The role of phosphatidylcholine in fatty acid exchange and desaturation in Brassica napus L. leaves.

    PubMed Central

    Williams, J P; Imperial, V; Khan, M U; Hodson, J N

    2000-01-01

    The role of phosphatidylcholine (PC) in fatty acid exchange and desaturation was examined and compared with that of monogalactosyldiacylglycerol (MGDG) in Brassica napus leaves using (14)C-labelling in vivo. Data are presented which indicate that in the chloroplast newly formed saturated (palmitic acid, 16:0) and monounsaturated (oleic acid, 18:1) fatty acid is incorporated into MGDG and desaturated in situ. In the non-plastidic compartments, however, newly formed fatty acid is exchanged with polyunsaturated fatty acid in PC, the probable major site of subsequent desaturation. The unsaturated fatty acid is released to the acyl-CoA pool, which is then used to synthesize diacylglycerol (DAG) containing a high level of unsaturated fatty acid. This highly unsaturated DAG may be the source for the biosynthesis of other cellular glycerolipids. The generally accepted pathway in which PC is synthesized from molecular species of DAG containing 16:0 and 18:1 followed by desaturation of the 18:1 to linoleic (18:2) and linolenic (18:3) acids is questioned. PMID:10861220

  8. Pertussis toxin treatment attenuates some effects of insulin in BC3H-1 murine myocytes

    SciTech Connect

    Luttrell, L.M.; Hewlett, E.L.; Romero, G.; Rogol, A.D.

    1988-05-05

    The effects of pertussis toxin (PT) treatment on insulin-stimulated myristoyl-diacylglycerol (DAG) generation, hexose transport, and thymidine incorporation were studied in differentiated BC3H-1 mycocytes. Insulin treatment caused a biphasic increase in myristoyl-DAG production which was abolished in myocytes treated with PT. There was no effect of PT treatment on basal (nonstimulated) myristoyl-DAG production. Insulin-stimulated hydrolysis of a membrane phosphatidylinositol glycan was blocked by PT treatment. ADP-ribosylation of BC3H-1 plasma membranes with (/sup 32/P)NAD revealed a 40-kDa protein as the major PT substrate in vivo and in vitro. The time course and dose dependence of the effects of PT on diacylglycerol generation correlated with the in vivo ADP-ribosylation of the 40-kDa substrate. Pertussis toxin treatment resulted in a 71% attenuation of insulin-stimulated hexose uptake without effect on either basal or phorbol ester-stimulated uptake. The stimulatory effects of insulin and fetal calf serum on (/sup 3/H)thymidine incorporation into quiescent myocytes were attenuated by 61 and 59%, respectively, when PT was added coincidently with the growth factors. Nonstimulated and EGF-stimulated (/sup 3/H)thymidine incorporation was unaffected by PT treatment. These data suggest that a PT-sensitive G protein is involved in the cellular signaling mechanisms of insulin.

  9. Disruption of the Arabidopsis Defense Regulator Genes SAG101, EDS1, and PAD4 Confers Enhanced Freezing Tolerance.

    PubMed

    Chen, Qin-Fang; Xu, Le; Tan, Wei-Juan; Chen, Liang; Qi, Hua; Xie, Li-Juan; Chen, Mo-Xian; Liu, Bin-Yi; Yu, Lu-Jun; Yao, Nan; Zhang, Jian-Hua; Shu, Wensheng; Xiao, Shi

    2015-10-01

    In Arabidopsis, three lipase-like regulators, SAG101, EDS1, and PAD4, act downstream of resistance protein-associated defense signaling. Although the roles of SAG101, EDS1, and PAD4 in biotic stress have been extensively studied, little is known about their functions in plant responses to abiotic stresses. Here, we show that SAG101, EDS1, and PAD4 are involved in the regulation of freezing tolerance in Arabidopsis. With or without cold acclimation, the sag101, eds1, and pad4 single mutants, as well as their double mutants, exhibited similarly enhanced tolerance to freezing temperatures. Upon cold exposure, the sag101, eds1, and pad4 mutants showed increased transcript levels of C-REPEAT/DRE BINDING FACTORs and their regulons compared with the wild type. Moreover, freezing-induced cell death and accumulation of hydrogen peroxide were ameliorated in sag101, eds1, and pad4 mutants. The sag101, eds1, and pad4 mutants had much lower salicylic acid (SA) and diacylglycerol (DAG) contents than the wild type, and exogenous application of SA and DAG compromised the freezing tolerance of the mutants. Furthermore, SA suppressed the cold-induced expression of DGATs and DGKs in the wild-type leaves. These findings indicate that SAG101, EDS1, and PAD4 are involved in the freezing response in Arabidopsis, at least in part, by modulating the homeostasis of SA and DAG. PMID:26149542

  10. Lipid phosphate phosphatase 3 participates in transport carrier formation and protein trafficking in the early secretory pathway.

    PubMed

    Gutiérrez-Martínez, Enric; Fernández-Ulibarri, Inés; Lázaro-Diéguez, Francisco; Johannes, Ludger; Pyne, Susan; Sarri, Elisabet; Egea, Gustavo

    2013-06-15

    The inhibition of phosphatidic acid phosphatase (PAP) activity by propanolol indicates that diacylglycerol (DAG) is required for the formation of transport carriers at the Golgi and for retrograde trafficking to the ER. Here we report that the PAP2 family member lipid phosphate phosphatase 3 (LPP3, also known as PAP2b) localizes in compartments of the secretory pathway from ER export sites to the Golgi complex. The depletion of human LPP3: (i) reduces the number of tubules generated from the ER-Golgi intermediate compartment and the Golgi, with those formed from the Golgi being longer in LPP3-silenced cells than in control cells; (ii) impairs the Rab6-dependent retrograde transport of Shiga toxin subunit B from the Golgi to the ER, but not the anterograde transport of VSV-G or ssDsRed; and (iii) induces a high accumulation of Golgi-associated membrane buds. LPP3 depletion also reduces levels of de novo synthesized DAG and the Golgi-associated DAG contents. Remarkably, overexpression of a catalytically inactive form of LPP3 mimics the effects of LPP3 knockdown on Rab6-dependent retrograde transport. We conclude that LPP3 participates in the formation of retrograde transport carriers at the ER-Golgi interface, where it transitorily cycles, and during its route to the plasma membrane. PMID:23591818