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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

    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.

  3. 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

  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

    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

  20. 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

  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. 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…

  4. 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

  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. 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.

  1. 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.

  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. 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

  12. 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...

  13. 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...

  14. 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...

  15. 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...

  16. 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...

  17. 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...

  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

    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

  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 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

  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. 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.

  7. 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

  8. 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

  9. 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

  10. 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

  11. 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

  12. 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

  13. 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

  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 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.

  16. 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.

  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. 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

  19. 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

  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. 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

  4. 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

  5. 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

  6. 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

  7. 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

  8. 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

  9. 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

  10. 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

  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. 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

  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. 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

  15. 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

  16. 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.

  17. 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

  18. 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.

  19. 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

  20. 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)

  1. 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)

  2. 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.

  3. 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)

  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. 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

  6. 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

  7. 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.

  8. [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

  9. 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

  10. 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

  11. 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

  12. 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

  13. 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

  14. 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

  15. 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

  16. 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

  17. 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.

  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. 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

  20. 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.

  1. 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.

  2. 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

  3. 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

  4. 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

  5. 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

  6. 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

  7. 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

  8. [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

  9. 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

  10. 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

  11. 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

  12. 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

  13. 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.

  14. 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

  15. 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

  16. 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.

  17. 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

  18. 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

  19. 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

  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. 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

  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. 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

  6. 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

  7. 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

  8. 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

  9. 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

  10. 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

  11. 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

  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. 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.

  17. 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

  18. 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.

  19. 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

  20. 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

  1. 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

  2. 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

  3. 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

  4. 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

  5. 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,…

  6. 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

  7. 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...

  8. 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...

  9. 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

  10. 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...

  11. 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

  12. 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)

  13. 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

  14. 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

  15. 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...

  16. 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.

  17. 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 ...

  18. 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...

  19. 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

  20. 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

  1. 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

  2. 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.

  3. 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

  4. 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

  5. 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...

  6. 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...

  7. 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

  8. 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

  9. 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

  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. 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. 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...

  13. 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…

  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. 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

  16. 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

  17. 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

  18. 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

  19. 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

  20. 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

  1. 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

  2. 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

  3. 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

  4. 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

  5. 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

  6. 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

  7. 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

  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. 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

  11. 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

  12. 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.

  13. 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

  14. 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

  15. 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.

  16. 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

  17. 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.

  18. 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

  19. 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

  20. 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.

  1. 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

  2. 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

  3. 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.

  4. 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

  5. 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

  6. 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

  7. 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

  8. 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.

  9. 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

  10. 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

  11. 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

  12. 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

  13. 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

  14. 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.

  15. 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

  16. 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

  17. 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

  18. 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

  19. 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

  20. 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.

  1. 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…

  2. 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

  3. 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

  4. 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.

  5. 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

  6. 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

  7. Choline-releasing glycerophosphodiesterase EDI3 links the tumor metabolome to signaling network activities.

    PubMed

    Marchan, Rosemarie; Lesjak, Michaela S; Stewart, Joanna D; Winter, Roland; Seeliger, Janine; Hengstler, Jan G

    2012-12-15

    Recently, EDI3 was identified as a key factor for choline metabolism that controls tumor cell migration and is associated with metastasis in endometrial carcinomas. EDI3 cleaves glycerophosphocholine (GPC) to form choline and glycerol-3-phosphate (G3P). Choline is then further metabolized to phosphatidylcholine (PtdC), the major lipid in membranes and a key player in membrane-mediated cell signaling. The second product, G3P, is a precursor molecule for several lipids with central roles in signaling, for example lysophosphatidic acid (LPA), phosphatidic acid (PA) and diacylglycerol (DAG). LPA activates intracellular signaling pathways by binding to specific LPA receptors, including membrane-bound G protein-coupled receptors and the intracellular nuclear receptor, PPARγ. Conversely, PA and DAG mediate signaling by acting as lipid anchors that bind and activate several signaling proteins. For example, binding of GTPases and PKC to PA and DAG, respectively, increases the activation of signaling networks, mediating processes such as migration, adhesion, proliferation or anti-apoptosis-all relevant for tumor development. We present a concept by which EDI3 either directly generates signaling molecules or provides "membrane anchors" for downstream signaling factors. As a result, EDI3 links choline metabolism to signaling activities resulting in a more malignant phenotype. PMID:23114620

  8. Bayesian networks as a tool for epidemiological systems analysis

    NASA Astrophysics Data System (ADS)

    Lewis, F. I.

    2012-11-01

    Bayesian network analysis is a form of probabilistic modeling which derives from empirical data a directed acyclic graph (DAG) describing the dependency structure between random variables. Bayesian networks are increasingly finding application in areas such as computational and systems biology, and more recently in epidemiological analyses. The key distinction between standard empirical modeling approaches, such as generalised linear modeling, and Bayesian network analyses is that the latter attempts not only to identify statistically associated variables, but to additionally, and empirically, separate these into those directly and indirectly dependent with one or more outcome variables. Such discrimination is vastly more ambitious but has the potential to reveal far more about key features of complex disease systems. Applying Bayesian network modeling to biological and medical data has considerable computational demands, combined with the need to ensure robust model selection given the vast model space of possible DAGs. These challenges require the use of approximation techniques, such as the Laplace approximation, Markov chain Monte Carlo simulation and parametric bootstrapping, along with computational parallelization. A case study in structure discovery - identification of an optimal DAG for given data - is presented which uses additive Bayesian networks to explore veterinary disease data of industrial and medical relevance.

  9. Age-associated changes of insulin action on the hydrolysis of diacylglycerol generated from phosphatidic acid.

    PubMed

    Pasquaré, S J; Salvador, G A; Giusto, N M

    2006-07-01

    Age-related changes in insulin action on diacylglycerol (DAG) degradation was studied in rat cerebral cortex synaptosomes. The generation of monoacylglycerol (MAG) and water soluble products (WSP, glycerol plus glycerol-3-phosphate) from DAG was studied in cerebral cortex (CC) synaptosomes from adult (4-month-old) and aged (28-month-old) rats. Additionally, the effect of porcine insulin and tyrosine phosphorylation was evaluated in the same group of animals. In this study we demonstrate that the age-related increase in WSP generation was accompanied by unmodified MAG levels. In the presence of diacylglycerol lipase (DAG lipase) inhibitor, RHC-80267, a lower inhibitory effect on MAG production was observed in CC synaptosomes from aged rats with respect to that in adult membranes. Under these experimental conditions, WSP formation was only diminished in aged membranes. Insulin stimulated MAG and WSP formation at long incubation times (30 min) in adult animals, while it had an inhibitory effect in aged animals. Insulin plus vanadate (as tyrosine-phosphatase inhibitor) inhibited MAG production at short incubation times whereas the same effect was observed in aged animals at long times of incubation. WSP formation was stimulated by insulin plus vanadate both in adult and aged animals at 30 min of incubation. Our results show that insulin differentially modulates MAG and WSP production from exogenous PA in CC synaptosomes from aged rats compared with adult rats. PMID:16651016

  10. Graft-union development: a delicate process that involves cell–cell communication between scion and stock for local auxin accumulation

    PubMed Central

    Yin, Hao; Yan, Bo; Sun, Jing; Jia, Pengfei; Zhang, Zijuan; Yan, Xiaosa; Chai, Juan; Ren, Zhizhong; Zheng, Guochang; Liu, Heng

    2012-01-01

    Grafting is an ancient cloning method that has been used widely for thousands of years in agricultural practices. Graft-union development is also an intricate process that involves substantial changes such as organ regeneration and genetic material exchange. However, the molecular mechanisms for graft-union development are still largely unknown. Here, a micrografting method that has been used widely in Arabidopsis was improved to adapt it a smooth procedure to facilitate sample analysis and to allow it to easily be applied to various dicotyledonous plants. The developmental stage of the graft union was characterized based on this method. Histological analysis suggested that the transport activities of vasculature were recovered at 3 days after grafting (dag) and that auxin modulated the vascular reconnection at 2 dag. Microarray data revealed a signal-exchange process between cells of the scion and stock at 1 dag, which re-established the communication network in the graft union. This process was concomitant with the clearing of cell debris, and both processes were initiated by a wound-induced programme. The results demonstrate the feasibility and potential power of investigating various plant developmental processes by this method, and represent a primary and significant step in interpretation of the molecular mechanisms underlying graft-union development. PMID:22511803

  11. NASA Langley and NLR Research of Distributed Air/Ground Traffic Management

    NASA Technical Reports Server (NTRS)

    Ballin, Mark G.; Hoekstra, Jacco M.; Wing, David J.; Lohr, Gary W.

    2002-01-01

    Distributed Air/Ground Traffic Management (DAG-TM) is a concept of future air traffic operations that proposes to distribute information, decision-making authority, and responsibility among flight crews, the air traffic service provider, and aeronautical operational control organizations. This paper provides an overview and status of DAG-TM research at NASA Langley Research Center and the National Aerospace Laboratory of The Netherlands. Specific objectives of the research are to evaluate the technical and operational feasibility of the autonomous airborne component of DAG-TM, which is founded on the operational paradigm of free flight. The paper includes an overview of research approaches, the airborne technologies under development, and a summary of experimental investigations and findings to date. Although research is not yet complete, these findings indicate that free flight is feasible and will significantly enhance system capacity and safety. While free flight cannot alone resolve the complex issues faced by those modernizing the global airspace, it should be considered an essential part of a comprehensive air traffic management modernization activity.

  12. RASA3 is a critical inhibitor of RAP1-dependent platelet activation

    PubMed Central

    Stefanini, Lucia; Paul, David S.; Robledo, Raymond F.; Chan, E. Ricky; Getz, Todd M.; Campbell, Robert A.; Kechele, Daniel O.; Casari, Caterina; Piatt, Raymond; Caron, Kathleen M.; Mackman, Nigel; Weyrich, Andrew S.; Parrott, Matthew C.; Boulaftali, Yacine; Adams, Mark D.; Peters, Luanne L.; Bergmeier, Wolfgang

    2015-01-01

    The small GTPase RAP1 is critical for platelet activation and thrombus formation. RAP1 activity in platelets is controlled by the GEF CalDAG-GEFI and an unknown regulator that operates downstream of the adenosine diphosphate (ADP) receptor, P2Y12, a target of antithrombotic therapy. Here, we provide evidence that the GAP, RASA3, inhibits platelet activation and provides a link between P2Y12 and activation of the RAP1 signaling pathway. In mice, reduced expression of RASA3 led to premature platelet activation and markedly reduced the life span of circulating platelets. The increased platelet turnover and the resulting thrombocytopenia were reversed by concomitant deletion of the gene encoding CalDAG-GEFI. Rasa3 mutant platelets were hyperresponsive to agonist stimulation, both in vitro and in vivo. Moreover, activation of Rasa3 mutant platelets occurred independently of ADP feedback signaling and was insensitive to inhibitors of P2Y12 or PI3 kinase. Together, our results indicate that RASA3 ensures that circulating platelets remain quiescent by restraining CalDAG-GEFI/RAP1 signaling and suggest that P2Y12 signaling is required to inhibit RASA3 and enable sustained RAP1-dependent platelet activation and thrombus formation at sites of vascular injury. These findings provide insight into the antithrombotic effect of P2Y12 inhibitors and may lead to improved diagnosis and treatment of platelet-related disorders. PMID:25705885

  13. 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.

  14. 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.

  15. 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

  16. Simultaneous production of fatty acid methyl esters and diglycerides by four recombinant Candida rugosa lipase's isozymes.

    PubMed

    Chang, Shu-Wei; Huang, Myron; Hsieh, Yu-Hsun; Luo, Ying-Ting; Wu, Tsung-Ta; Tsai, Chia-Wen; Chen, Chin-Shuh; Shaw, Jei-Fu

    2014-07-15

    In this study, the catalytic efficiency of four recombinant CRL (Candida rugosa lipase) isozymes (LIP1-LIP4) towards the production of fatty acid methyl ester (FAME) was compared and evaluated as an alternative green method for industrial applications. The results indicated that the recombinant C. rugosa LIP1 enzyme exhibited the highest catalytic efficiency for FAME production compared to the recombinant C. rugosa LIP2-LIP4 enzymes. The optimal conditions were as follows: pH 7.0, methanol/soybean oil molar ratio: 3/1, enzyme amount: 2U (1.6 μL), reaction temperature: 20°C, 22 h of reaction time, and 3 times of methanol addition (1 mol/6h), and resulted in 61.5 ± 1.5 wt.% of FAME conversion. The reaction product contained also 10 wt.% of DAG with a ratio of 1,3-DAG to 1,2-DAG of approximately 4:6, and can be potentially used in industrial applications as a food emulsifier. PMID:24594166

  17. Causal Analysis After Haavelmo

    PubMed Central

    Heckman, James; Pinto, Rodrigo

    2014-01-01

    Haavelmo's seminal 1943 and 1944 papers are the first rigorous treatment of causality. In them, he distinguished the definition of causal parameters from their identification. He showed that causal parameters are defined using hypothetical models that assign variation to some of the inputs determining outcomes while holding all other inputs fixed. He thus formalized and made operational Marshall's (1890) ceteris paribus analysis. We embed Haavelmo's framework into the recursive framework of Directed Acyclic Graphs (DAGs) used in one influential recent approach to causality (Pearl, 2000) and in the related literature on Bayesian nets (Lauritzen, 1996). We compare the simplicity of an analysis of causality based on Haavelmo's methodology with the complex and nonintuitive approach used in the causal literature of DAGs—the “do-calculus” of Pearl (2009). We discuss the severe limitations of DAGs and in particular of the do-calculus of Pearl in securing identification of economic models. We extend our framework to consider models for simultaneous causality, a central contribution of Haavelmo. In general cases, DAGs cannot be used to analyze models for simultaneous causality, but Haavelmo's approach naturally generalizes to cover them. PMID:25729123

  18. Lipid Droplets Form from Distinct Regions of the Cell in the Fission Yeast Schizosaccharomyces pombe.

    PubMed

    Meyers, Alex; Del Rio, Zuania P; Beaver, Rachael A; Morris, Ryan M; Weiskittel, Taylor M; Alshibli, Amany K; Mannik, Jaana; Morrell-Falvey, Jennifer; Dalhaimer, Paul

    2016-06-01

    Eukaryotic cells store cholesterol/sterol esters (SEs) and triacylglycerols (TAGs) in lipid droplets, which form from the contiguous endoplasmic reticulum (ER) network. However, it is not known if droplets preferentially form from certain regions of the ER over others. Here, we used fission yeast Schizosaccharomyces pombe cells where the nuclear and cortical/peripheral ER domains are distinguishable by light microscopy to show that SE-enriched lipid droplets form away from the nucleus at the cell tips, whereas TAG-enriched lipid droplets form around the nucleus. Sterols localize to the regions of the cells where droplets enriched in SEs are observed. TAG droplet formation around the nucleus appears to be a strong function of diacylglycerol (DAG) homeostasis with Cpt1p, which coverts DAG into phosphatidylcholine and phosphatidylethanolamine localized exclusively to the nuclear ER. Also, Dgk1p, which converts DAG into phosphatidic acid localized strongly to the nuclear ER over the cortical/peripheral ER. We also show that TAG more readily translocates from the ER to lipid droplets than do SEs. The results augment the standard lipid droplet formation model, which has SEs and TAGs flowing into the same nascent lipid droplet regardless of its biogenesis point in the cell. PMID:26990381

  19. Effective diagnosis of genetic disease by computational phenotype analysis of the disease-associated genome

    PubMed Central

    Zemojtel, Tomasz; Köhler, Sebastian; Mackenroth, Luisa; Jäger, Marten; Hecht, Jochen; Krawitz, Peter; Graul-Neumann, Luitgard; Doelken, Sandra; Ehmke, Nadja; Spielmann, Malte; Øien, Nancy Christine; Schweiger, Michal R.; Krüger, Ulrike; Frommer, Götz; Fischer, Björn; Kornak, Uwe; Flöttmann, Ricarda; Ardeshirdavani, Amin; Moreau, Yves; Lewis, Suzanna E.; Haendel, Melissa; Smedley, Damian; Horn, Denise; Mundlos, Stefan; Robinson, Peter N.

    2015-01-01

    Less than half of patients with suspected genetic disease receive a molecular diagnosis. We have therefore integrated next-generation sequencing (NGS), bioinformatics, and clinical data into an effective diagnostic workflow. We used variants in the 2741 established Mendelian disease genes [the disease-associated genome (DAG)] to develop a targeted enrichment DAG panel (7.1 Mb), which achieves a coverage of 20-fold or better for 98% of bases. Furthermore, we established a computational method [Phenotypic Interpretation of eXomes (PhenIX)] that evaluated and ranked variants based on pathogenicity and semantic similarity of patients’ phenotype described by Human Phenotype Ontology (HPO) terms to those of 3991 Mendelian diseases. In computer simulations, ranking genes based on the variant score put the true gene in first place less than 5% of the time; PhenIX placed the correct gene in first place more than 86% of the time. In a retrospective test of PhenIX on 52 patients with previously identified mutations and known diagnoses, the correct gene achieved a mean rank of 2.1. In a prospective study on 40 individuals without a diagnosis, PhenIX analysis enabled a diagnosis in 11 cases (28%, at a mean rank of 2.4). Thus, the NGS of the DAG followed by phenotype-driven bioinformatic analysis allows quick and effective differential diagnostics in medical genetics. PMID:25186178

  20. Lipidomic analysis of Arabidopsis seed genetically engineered to contain DHA

    PubMed Central

    Zhou, Xue-Rong; Callahan, Damien L.; Shrestha, Pushkar; Liu, Qing; Petrie, James R.; Singh, Surinder P.

    2014-01-01

    Metabolic engineering of omega-3 long-chain (≥C20) polyunsaturated fatty acids (ω3 LC-PUFA) in oilseeds has been one of the key targets in recent years. By expressing a transgenic pathway for enhancing the synthesis of the ω3 LC-PUFA docosahexaenoic acid (DHA) from endogenous α-linolenic acid (ALA), we obtained the production of fish oil-like proportions of DHA in Arabidopsis seed oil. Liquid chromatography-mass spectrometry (LC-MS) was used to characterize the triacylglycerol (TAG), diacylglycerol (DAG) and phospholipid (PL) lipid classes in the transgenic and wild type Arabidopsis seeds at both developing and mature stages. The analysis identified the appearance of several abundant DHA-containing phosphatidylcholine (PC), DAG and TAG molecular species in mature seeds. The relative abundances of PL, DAG, and TAG species showed a preferred combination of LC-PUFA with ALA in the transgenic seeds, where LC-PUFA were esterified in positions usually occupied by 20:1ω9. Trace amounts of di-DHA PC and tri-DHA TAG were identified and confirmed by high resolution MS/MS. Studying the lipidome in transgenic seeds provided insights into where DHA accumulated and combined with other fatty acids of neutral and phospholipids from the developing and mature seeds. PMID:25225497

  1. Mandatory leaf node prediction in hierarchical multilabel classification.

    PubMed

    Bi, Wei; Kwok, James T

    2014-12-01

    In hierarchical classification, the output labels reside on a tree- or directed acyclic graph (DAG)-structured hierarchy. On testing, the prediction paths of a given test example may be required to end at leaf nodes of the label hierarchy. This is called mandatory leaf node prediction (MLNP) and is particularly useful, when the leaf nodes have much stronger semantic meaning than the internal nodes. However, while there have been a lot of MLNP methods in hierarchical multiclass classification, performing MLNP in hierarchical multilabel classification is difficult. In this paper, we propose novel MLNP algorithms that consider the global label hierarchy structure. We show that the joint posterior probability over all the node labels can be efficiently maximized by dynamic programming for label trees, or greedy algorithm for label DAGs. In addition, both algorithms can be further extended for the minimization of the expected symmetric loss. Experiments are performed on real-world MLNP data sets with label trees and label DAGs. The proposed method consistently outperforms other hierarchical and flat multilabel classification methods. PMID:25420248

  2. Adhalin, the 50 kD dystrophin associated protein, is not the locus for severe childhood autosomal recessive dystrophy (SCARMD)

    SciTech Connect

    McNally, E.M.; Selig, S.; Kunkel, L.M.

    1994-09-01

    Mutations in the carboxyl-terminus in dystrophin are normally sufficient to produce severely dystrophic muscle. This portion of dystrophin binds a complex of dystrophin-associated glycoproteins (DAGs). The genes encoding these DAGs are candidate genes for causing neuromuscular disease. Immunoreactivity for adhalin, the 50 kD DAG, is absent in muscle biopsies from patients with SCARMD, a form of dystrophy clinically similar Duchenne muscular dystrophy. Prior linkage analysis in SCARMD families revealed that the disease gene segregates with markers on chromosome 13. To determine the molecular role that adhalin may play in SCARMD, human cDNA and genomic sequences were isolated. Primers were designed based on predicted areas of conservation in rabbit adhalin and used in RT-PCR with human skeletal and cardiac muscle. RT-PCR products were confirmed by sequence as human adhalin and then used as probes for screening human cDNA and genomic libraries. Human and rabbit adhalin are 90% identical, and among the cDNAs, a novel splice form of adhalin was seen which may encode part of the 35 kD component of the dystrophin-glycoprotein complex. To our surprise, only human/rodent hybrids containing human chromosome 17 amplified adhalin sequences in a PCR analysis. FISH analysis with three overlapping genomic sequences confirmed the chromosome 17 location and further delineated the map position to 17q21. Therefore, adhalin is excluded as the gene causing SCARMD.

  3. 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

  4. Upregulation of myocellular DGAT1 augments triglyceride synthesis in skeletal muscle and protects against fat-induced insulin resistance

    PubMed Central

    Liu, Li; Zhang, Yiying; Chen, Nancy; Shi, Xiaojing; Tsang, Bonny; Yu, Yi-Hao

    2007-01-01

    Increased fat deposition in skeletal muscle is associated with insulin resistance. However, exercise increases both intramyocellular fat stores and insulin sensitivity, a phenomenon referred to as “the athlete’s paradox”. In this study, we provide evidence that augmenting triglyceride synthesis in skeletal muscle is intrinsically connected with increased insulin sensitivity. Exercise increased diacylglycerol (DAG) acyltransferase (DGAT) activity in skeletal muscle. Channeling fatty acid substrates into TG resulted in decreased DAG and ceramide levels. Transgenic overexpression of DGAT1 in mouse skeletal muscle replicated these findings and protected mice against high-fat diet–induced insulin resistance. Moreover, in isolated muscle, DGAT1 deficiency exacerbated insulin resistance caused by fatty acids, whereas DGAT1 overexpression mitigated the detrimental effect of fatty acids. The heightened insulin sensitivity in the transgenic mice was associated with attenuated fat-induced activation of DAG-responsive PKCs and the stress mediator JNK1. Consistent with these changes, serine phosphorylation of insulin receptor substrate 1 was reduced, and Akt activation and glucose 4 membrane translocation were increased. In conclusion, upregulation of DGAT1 in skeletal muscle is sufficient to recreate the athlete’s paradox and illustrates a mechanism of exercise-induced enhancement of muscle insulin sensitivity. Thus, increasing muscle DGAT activity may offer a new approach to prevent and treat insulin resistance and type 2 diabetes mellitus. PMID:17510710

  5. Classical transient receptor potential channel 6 (TRPC6) is essential for hypoxic pulmonary vasoconstriction and alveolar gas exchange

    PubMed Central

    Weissmann, Norbert; Dietrich, Alexander; Fuchs, Beate; Kalwa, Hermann; Ay, Mahmut; Dumitrascu, Rio; Olschewski, Andrea; Storch, Ursula; Mederos y Schnitzler, Michael; Ghofrani, Hossein Ardeschir; Schermuly, Ralph Theo; Pinkenburg, Olaf; Seeger, Werner; Grimminger, Friedrich; Gudermann, Thomas

    2006-01-01

    Regional alveolar hypoxia causes local vasoconstriction in the lung, shifting blood flow from hypoxic to normoxic areas, thereby maintaining gas exchange. This mechanism is known as hypoxic pulmonary vasoconstriction (HPV). Disturbances in HPV can cause life-threatening hypoxemia whereas chronic hypoxia triggers lung vascular remodeling and pulmonary hypertension. The signaling cascade of this vitally important mechanism is still unresolved. Using transient receptor potential channel 6 (TRPC6)-deficient mice, we show that this channel is a key regulator of acute HPV as this regulatory mechanism was absent in TRPC6−/− mice whereas the pulmonary vasoconstrictor response to the thromboxane mimetic U46619 was unchanged. Accordingly, induction of regional hypoventilation resulted in severe arterial hypoxemia in TRPC6−/− but not in WT mice. This effect was mirrored by a lack of hypoxia-induced cation influx and currents in smooth-muscle cells from precapillary pulmonary arteries (PASMC) of TRPC6−/− mice. In both WT and TRPC6−/− PASMC hypoxia caused diacylglycerol (DAG) accumulation. DAG seems to exert its action via TRPC6, as DAG kinase inhibition provoked a cation influx only in WT but not in TRPC6−/− PASMC. Notably, chronic hypoxia-induced pulmonary hypertension was independent of TRPC6 activity. We conclude that TRPC6 plays a unique and indispensable role in acute hypoxic pulmonary vasoconstriction. Manipulation of TRPC6 function may thus offer a therapeutic strategy for the control of pulmonary hemodynamics and gas exchange. PMID:17142322

  6. Involvement of second messengers in regulation of the low-density lipoprotein receptor gene.

    PubMed Central

    Auwerx, J H; Chait, A; Wolfbauer, G; Deeb, S S

    1989-01-01

    Transcription of the low-density lipoprotein receptor (LDL-R) gene in the human monocytic leukemic cell line THP-1 and in the human hepatocarcinoma cell line Hep-G2 is regulated by second messengers of the diacylglycerol-protein kinase C (DAG-PKC), inositol 1,4,5-triphosphate-Ca2+, and cyclic AMP pathways. Exogenous phospholipase C (which releases DAG and inositol 1,4,5-triphosphate), PKC activators (phorbol esters and DAG), Ca2+ ionophores, and a cyclic AMP analog all transiently induced accumulation of LDL-R mRNA. The effects of these three signal-transducing pathways were to a large extent additive. Furthermore, PKC stimulation effected an increase in LDL binding, which suggested that the increase in LDL-R mRNA resulted in an increase in functional cell surface receptor activity. These results suggest that uptake of cholesterol by these cells is under control of both intracellular cholesterol levels and external signals. Images PMID:2548077

  7. Use of re-esterified oils, differing in their degree of saturation and molecular structure, in broiler chicken diets.

    PubMed

    Vilarrasa, E; Codony, R; Esteve-Garcia, E; Barroeta, A C

    2015-07-01

    The aim of the present study was to investigate the potential use of re-esterified oils, differing in their degree of saturation and molecular structure, in comparison with their corresponding acid and native oils in broiler chicken diets. For this purpose, 144 one-d-old female broiler chickens were randomly distributed in 48 cages. Birds were fed a basal diet supplemented with 6% of native palm oil ( PN: ), acid palm oil ( PA: ), re-esterified palm oil low in mono- ( MAG: ) and diacylglycerols ( DAG: ) ( PEL: ), re-esterified palm oil high in MAG and DAG ( PEH: ), native soybean oil ( SN: ), acid soybean oil ( SA: ), re-esterified soybean oil low in MAG and DAG ( SEL: ), or re-esterified soybean oil high in MAG and DAG ( SEH: ), which resulted in a 2 × 4 factorial arrangement. Digestibility balances showed that the degree of saturation of fat generally exerted a greater impact than did the fat molecular structure. The dietary utilization of S sources was higher than that of P sources. However, the increased sn-2 saturated fatty acid ( SFA: ) content of EL oils in the starter period and the increased MAG and DAG content of EH oils in the grower-finisher period yielded favorable effects on the SFA apparent absorption, especially in those birds fed re-esterified palm oils. The excreta acylglycerol and free fatty acid composition was mainly composed of free fatty acids, and their amount almost paralleled the results observed for SFA apparent absorption. For growth performance, birds fed S exhibited better feed conversion ratios and lower abdominal fat-pad weights than did those fed P. The fatty acid composition of abdominal adipose tissue was also mainly affected by the degree of saturation of dietary fat sources. We concluded that re-esterified oils, mainly from P sources, can be used in broiler chicken diets as alternative fat sources since they show similar or even higher total fatty acid apparent absorption than do their corresponding native and acid oils, with

  8. Phospholipase C beta3 is a key component in the Gbetagamma/PKCeta/PKD-mediated regulation of trans-Golgi network to plasma membrane transport.

    PubMed

    Díaz Añel, Alberto M

    2007-08-15

    The requirement of DAG (diacylglycerol) to recruit PKD (protein kinase D) to the TGN (trans-Golgi network) for the targeting of transport carriers to the cell surface, has led us to a search for new components involved in this regulatory pathway. Previous findings reveal that the heterotrimeric Gbetagamma (GTP-binding protein betagamma subunits) act as PKD activators, leading to fission of transport vesicles at the TGN. We have recently shown that PKCeta (protein kinase Ceta) functions as an intermediate member in the vesicle generating pathway. DAG is capable of activating this kinase at the TGN, and at the same time is able to recruit PKD to this organelle in order to interact with PKCeta, allowing phosphorylation of PKD's activation loop. The most qualified candidates for the production of DAG at the TGN are PI-PLCs (phosphatidylinositol-specific phospholipases C), since some members of this family can be directly activated by Gbetagamma, utilizing PtdIns(4,5)P2 as a substrate, to produce the second messengers DAG and InsP3. In the present study we show that betagamma-dependent Golgi fragmentation, PKD1 activation and TGN to plasma membrane transport were affected by a specific PI-PLC inhibitor, U73122 [1-(6-{[17-3-methoxyestra-1,3,5(10)-trien-17-yl]amino}hexyl)-1H-pyrrole-2,5-dione]. In addition, a recently described PI-PLC activator, m-3M3FBS [2,4,6-trimethyl-N-(m-3-trifluoromethylphenyl)benzenesulfonamide], induced vesiculation of the Golgi apparatus as well as PKD1 phosphorylation at its activation loop. Finally, using siRNA (small interfering RNA) to block several PI-PLCs, we were able to identify PLCbeta3 as the sole member of this family involved in the regulation of the formation of transport carriers at the TGN. In conclusion, we demonstrate that fission of transport carriers at the TGN is dependent on PI-PLCs, specifically PLCbeta3, which is necessary to activate PKCeta and PKD in that Golgi compartment, via DAG production. PMID:17492941

  9. Cell signaling and estrogens in female rat osteoblasts: a possible involvement of unconventional nonnuclear receptors.

    PubMed

    Lieberherr, M; Grosse, B; Kachkache, M; Balsan, S

    1993-11-01

    Estrogen deficiency is associated with bone loss, and estrogen replacement is an effective treatment of this osteoporotic process. This study examines the early (5-120 s) effects of 17 beta-estradiol on the intracellular calcium and phospholipid metabolism in confluent female rat osteoblasts. The cytosolic free Ca2+ concentration ([Ca2+]i) was determined using fura-2/AM as Ca2+ probe. Cells were labeled with myo-[2-3H]inositol or [14C]arachidonic acid for inositol or lipid determination. Inositol 1,4,5-trisphosphate (IP3) and diacylglycerol (DAG) production were determined by either mass measurement or anion-exchange chromatography or by thin-layer chromatography, respectively. 17 beta-Estradiol (1 pM to 1 nM) increased [Ca2+]i in a biphasic manner within 10 s via Ca2+ influx from the extracellular milieu, as shown by the effects of the calcium chelator EGTA and the Ca2+ channel blockers nifedipine and verapamil, and via Ca2+ mobilization from the endoplasmic reticulum (ER), as shown by the effects of thapsigargin. 17 beta-Estradiol (1 pM to 1 nM) induced a biphasic and concomitant increase in IP3 and DAG formation. Estradiol immobilized on bovine serum albumin (BSA) [E-(O-carboxymethyl)oxime BSA] and its derivative (O-carboxymethyl)oxime rapidly increased ([Ca2+]i, IP3, and DAG and were full agonists, although they were less potent than the free estradiol. They had the same action time course and acted via Ca2+ influx and Ca2+ mobilization from ER. Tamoxifen, a potent inhibitor of genomic steroid responses, did not block the rapid increase in Ca2+, IP3, and DAG induced by estradiol. Finally, inhibitor of phospholipase C (neomycin) and pertussis toxin abolished the effects of 17 beta-estradiol on IP3 and DAG formation. These results suggest that female rat osteoblasts bear non-genomic unconventional cell surface receptors for estradiol, belonging to the class of the membrane receptors coupled to a phospholipase C via a pertussis toxin-sensitive G protein. PMID

  10. Carl Ludwig: the man, his time, his influence.

    PubMed

    Zimmer, H G

    1996-01-01

    Carl Ludwig (1816-1895) was the driving force in the foundation and development of scientifically based and experimentally oriented physiology against natural philosophy and vitalism that prevailed during the first quarter of the 19th century in Germany. He was the representative of a small group of young, highly talented and dynamic physiologists aiming at implementing the laws of physics and chemistry as the only active forces in physiologic processes. These "organic physicists" included Emil du Bois-Reymond (1818-1896), Ernst Brücke (1819-1892), and Hermann Helmholz (1821-1894). Carl Ludwig wrote the program of this group in the form of a textbook of physiology that was considered revolutionary, provocative and premature. His academic life, his inventions and discoveries, his scientific achievements, his influence and his personality are reviewed. Since every person can be viewed only in the context of his time, the political background, the economic and social situation, the conditions for science and research as well as the cultural climate that were characteristic for the decisive years of Carl Ludwig are described to some extent. It is shown that Carl Ludwig and his contemporary organic physicists lived and grew into a science- and research-oriented period which had been prepared and paved by men like Johannes Evangelista Purkinje (1787-1869), Ernst Heinrich Weber (1795-1878), Alfred Wilhelm Volkmann (1800-1877), Johannes Müller (1801-1858) and Gustav Theodor Fechner (1801-1887). They benefited from this enormous scientific development and contributed to it to a large and significant extent so that it ultimately turned out to be the most productive and influential period in the history of German physiology. Some of the numerous scholars who had studied with Carl Ludwig carried his approach to physiology into the 20th century: Adolf Fick (1829-1901), Otto Frank (1865-1944), Iwan Petrowitsch Pawlow (1849-1936) and Henry Pickering Browditch (1840-1911). PMID

  11. Extensive impact melting on the H-chondrite parent asteroid during the cataclysmic bombardment of the early solar system: Evidence from the achondritic meteorite Dar al Gani 896

    NASA Astrophysics Data System (ADS)

    Folco, Luigi; Bland, Philip A.; D'Orazio, Massimo; Franchi, Ian A.; Kelley, Simon P.; Rocchi, Sergio

    2004-05-01

    DaG 896 is an olivine-rich microporphyritic rock of komatiitic composition. Both the olivine composition (Fa 17.5±2.1, [Mn/Mg] = 0.0061) and the bulk oxygen isotopic composition (δ 17O = +2.55, δ 18O = +3.50) indicate that DaG 896 is a sample of the H-chondrite parent body. The bulk chemistry shows an H-chondritic distribution of lithophile elements, whereas chalcophile and siderophile elements are strongly depleted, indicating formation through whole-rock melting (or nearly so) of H-chondrite material, nearly complete loss of the metal plus sulfide component, and crystallization without significant igneous fractionation. Superheated, severely shocked chondritic relics (˜10 vol%), typically in the form of corroded lithic fragments <100 μm in size intimately distributed within the igneous lithology, indicate that melting was triggered by a highly energetic impact, which possibly induced shock pressures of ˜80-100 GPa. The relatively young 3.704 ± 0.035 Ga 40Ar- 39Ar crystallization age is consistent with the impact melting origin, as magmatism in the asteroid belt was active only in the first hundred million years of solar system history. Based on textural data and thermodynamic crystallization modelling, we infer that DaG 896 crystallized from a liquidus temperature of ˜1630°C under relatively slow cooling rates (˜10°C h -1) to ˜1300°C, before quenching. The two-stage cooling history indicates that a reasonable formation environment might be a dike intruding cooler basement below a crater floor. Metal-silicate fractionation may have been accomplished, at least at the centimeter-scale of the studied meteorite sample, through differential acceleration of immiscible liquids of different density during the intense flow regimes associated with the excavation and modification stages of the cratering mechanism. Alternatively, DaG 896 may represent a surface sample of a differentiated melt body at the floor of an impact crater, as gravitational settling

  12. Inhibitory effects of herbal constituents on P-glycoprotein in vitro and in vivo: Herb–drug interactions mediated via P-gp

    SciTech Connect

    Li, Xue Hu, Jinping Wang, Baolian Sheng, Li Liu, Zhihao Yang, Shuang Li, Yan

    2014-03-01

    Modulation of drug transporters via herbal medicines which have been widely used in combination with conventional prescription drugs may result in herb–drug interactions in clinical practice. The present study was designed to investigate the inhibitory effects of 50 major herbal constituents on P-glycoprotein (P-gp) in vitro and in vivo as well as related inhibitory mechanisms. Among these herbal medicines, four constituents, including emodin, 18β-glycyrrhetic acid (18β-GA), dehydroandrographolide (DAG), and 20(S)-ginsenoside F{sub 1} [20(S)-GF{sub 1}] exhibited significant inhibition (> 50%) on P-gp in MDR1-MDCKII and Caco-2 cells. Emodin was the strongest inhibitor of P-gp (IC{sub 50} = 9.42 μM), followed by 18β-GA (IC{sub 50} = 21.78 μM), 20(S)-GF{sub 1} (IC{sub 50} = 76.08 μM) and DAG (IC{sub 50} = 77.80 μM). P-gp ATPase activity, which was used to evaluate the affinity of substrates to P-gp, was stimulated by emodin and DAG with K{sub m} and V{sub max} values of 48.61, 29.09 μM and 71.29, 38.45 nmol/min/mg protein, respectively. However, 18β-GA and 20(S)-GF{sub 1} exhibited significant inhibition on both basal and verapamil-stimulated P-gp ATPase activities at high concentration. Molecular docking analysis (CDOCKER) further elucidated the mechanism for structure–inhibition relationships of herbal constituents with P-gp. When digoxin was co-administered to male SD rats with emodin or 18β-GA, the AUC{sub 0−t} and Cmax of digoxin were increased by approximately 51% and 58%, respectively. Furthermore, 18β-GA, DAG, 20(S)-GF{sub 1} and Rh{sub 1} at 10 μM significantly inhibited CYP3A4/5 activity, while emodin activated the metabolism of midazolam in human liver microsomes. In conclusion, four herbal constituents demonstrated inhibition of P-gp to specific extents in vitro and in vivo. Taken together, our findings provided the basis for the reliable assessment of the potential risks of herb–drug interactions in humans. - Highlights: • Emodin, 18

  13. Characterization of the Rac-GAP (Rac-GTPase-activating protein) activity of beta2-chimaerin, a 'non-protein kinase C' phorbol ester receptor.

    PubMed Central

    Caloca, Maria Jose; Wang, HongBin; Kazanietz, Marcelo G

    2003-01-01

    The regulation and function of beta2-chimaerin, a novel receptor for the phorbol ester tumour promoters and the second messenger DAG (diacylglycerol), is largely unknown. As with PKC (protein kinase C) isoenzymes, phorbol esters bind to beta2-chimaerin with high affinity and promote its subcellular distribution. beta2-Chimaerin has GAP (GTPase-activating protein) activity for the small GTP-binding protein Rac1, but for not Cdc42 or RhoA. We show that acidic phospholipids enhanced its catalytic activity markedly in vitro, but the phorbol ester PMA had no effect. beta2-Chimaerin and other chimaerin isoforms decreased cellular levels of Rac-GTP markedly in COS-1 cells and impaired GTP loading on to Rac upon EGF (epidermal growth factor) receptor stimulation. Deletional and mutagenesis analysis determined that the beta2-chimaerin GAP domain is essential for this effect. Interestingly, PMA has a dual effect on Rac-GTP levels in COS-1 cells. PMA increased Rac-GTP levels in the absence of a PKC inhibitor, whereas under conditions in which PKC activity is inhibited, PMA markedly decreased Rac-GTP levels and potentiated the effect of beta2-chimaerin. Chimaerin isoforms co-localize at the plasma membrane with active Rac, and these results were substantiated by co-immunoprecipitation assays. In summary, the novel phorbol ester receptor beta2-chimaerin regulates the activity of the Rac GTPase through its GAP domain, leading to Rac inactivation. These results strongly emphasize the high complexity of DAG signalling due to the activation of PKC-independent pathways, and cast doubts regarding the selectivity of phorbol esters and DAG analogues as selective PKC activators. PMID:12877655

  14. Diacylglycerol kinase θ: regulation and stability.

    PubMed

    Tu-Sekine, Becky; Goldschmidt, Hana; Petro, Elizabeth; Raben, Daniel M

    2013-01-01

    Given the well-established roles of diacylglycerol (DAG) and phosphatidic acid (PtdOH) in a variety of signaling cascades, it is not surprising that there is an increasing interest in understanding their physiological roles and mechanisms that regulate their cellular levels. One class of enzymes capable of coordinately regulating the levels of these two lipids is the diacylglycerol kinases (DGKs). These enzymes catalyze the transfer of the γ-phosphate of ATP to the hydroxyl group of DAG, which generates PtdOH while reducing DAG. As these enzymes reciprocally modulate the relative levels of these two signaling lipids, it is essential to understand the regulation and roles of these enzymes in various tissues. One system where these enzymes play important roles is the nervous system. Of the ten mammalian DGKs, eight of them are readily detected in the mammalian central nervous system (CNS): DGK-α, DGK-β, DGK-γ, DGK-η, DGK-ζ, DGK-ι, DGK-ε, and DGK-θ. Despite the increasing interest in DGKs, little is known about their regulation. We have focused some attention on understanding the enzymology and regulation of one of these DGK isoforms, DGK-θ. We recently showed that DGK-θ is regulated by an accessory protein containing polybasic regions. We now report that this accessory protein is required for the previously reported broadening of the pH profile observed in cell lysates in response to phosphatidylserine (PtdSer). Our data further reveal DGK-θ is regulated by magnesium and zinc, and sensitive to the known DGK inhibitor R599022. These data outline new parameters involved in regulating DGK-θ. PMID:23266086

  15. Phospholipase C and D regulation of Src, calcium release and membrane fusion during Xenopus laevis development.

    PubMed

    Stith, Bradley J

    2015-05-15

    This review emphasizes how lipids regulate membrane fusion and the proteins involved in three developmental stages: oocyte maturation to the fertilizable egg, fertilization and during first cleavage. Decades of work show that phosphatidic acid (PA) releases intracellular calcium, and recent work shows that the lipid can activate Src tyrosine kinase or phospholipase C during Xenopus fertilization. Numerous reports are summarized to show three levels of increase in lipid second messengers inositol 1,4,5-trisphosphate and sn 1,2-diacylglycerol (DAG) during the three different developmental stages. In addition, possible roles for PA, ceramide, lysophosphatidylcholine, plasmalogens, phosphatidylinositol 4-phosphate, phosphatidylinositol 5-phosphate, phosphatidylinositol 4,5-bisphosphate, membrane microdomains (rafts) and phosphatidylinositol 3,4,5-trisphosphate in regulation of membrane fusion (acrosome reaction, sperm-egg fusion, cortical granule exocytosis), inositol 1,4,5-trisphosphate receptors, and calcium release are discussed. The role of six lipases involved in generating putative lipid second messengers during fertilization is also discussed: phospholipase D, autotaxin, lipin1, sphingomyelinase, phospholipase C, and phospholipase A2. More specifically, proteins involved in developmental events and their regulation through lipid binding to SH3, SH4, PH, PX, or C2 protein domains is emphasized. New models are presented for PA activation of Src (through SH3, SH4 and a unique domain), that this may be why the SH2 domain of PLCγ is not required for Xenopus fertilization, PA activation of phospholipase C, a role for PA during the calcium wave after fertilization, and that calcium/calmodulin may be responsible for the loss of Src from rafts after fertilization. Also discussed is that the large DAG increase during fertilization derives from phospholipase D production of PA and lipin dephosphorylation to DAG. PMID:25748412

  16. Calcium influx, but not intracellular calcium release, supports PACAP-mediated ERK activation in HEK PAC1 receptor cells.

    PubMed

    May, Victor; Clason, Todd A; Buttolph, Thomas R; Girard, Beatrice M; Parsons, Rodney L

    2014-11-01

    In HEK cells expressing GFP-tagged PAC1Hop1 receptors, PACAP augments ERK phosphorylation through two parallel pathways: one through PACAP/PAC1 receptor internalization/endosome MEK/ERK signaling and the other through PLC/DAG/PKC activation. We examined whether elevation of intracellular calcium ([Ca(2+)]i) was required for either of the PACAP/PAC1 receptor-mediated ERK activation mechanisms. The PACAP (25 nM)-induced elevation of [Ca(2+)]i was greater with cells maintained in Ca(2+)-containing than in Ca(2+)-deficient solution, suggesting that both calcium release from internal stores and calcium influx contributed to the rise in [Ca(2+)]i. A thapsigargin-induced increase in [Ca(2+)]i also was greater with calcium in the external solution. OAG, the cell permeable analogue of DAG, increased [Ca(2+)]i, but only in Ca(2+)-containing solution. Decreasing external calcium or depleting internal calcium stores did not block PACAP-induced PAC1 receptor internalization. Omission of calcium from the external solution, but not thapsigargin pretreatment, significantly blunted PACAP-stimulated ERK phosphorylation. The PKC inhibitor BimI decreased PACAP-mediated ERK activation in both Ca(2+)-containing or Ca(2+)-deficient solutions. In contrast, following Pitstop 2 pretreatment to block endocytic mechanisms, PACAP activated ERK only when calcium was present in the external solution. We conclude that the endosome signaling pathway is largely calcium-independent whereas calcium influx appears necessary for the PLC/DAG/PKC component of PACAP-induced ERK activation. PMID:24723666

  17. Phospholipase C and D regulation of Src, calcium release and membrane fusion during Xenopus laevis development

    PubMed Central

    Stith, Bradley J.

    2015-01-01

    This review emphasizes how lipids regulate membrane fusion and the proteins involved in three developmental stages: oocyte maturation to the fertilizable egg, fertilization and during first cleavage. Decades of work show that phosphatidic acid (PA) releases intracellular calcium, and recent work shows that the lipid can activate Src tyrosine kinase or phospholipase C during Xenopus fertilization. Numerous reports are summarized to show three levels of increase in lipid second messengers inositol 1,4,5-trisphosphate and sn 1,2-diacylglycerol (DAG) during the three different developmental stages. In addition, possible roles for PA, ceramide, lysophosphatidylcholine, plasmalogens, phosphatidylinositol 4-phosphate, phosphatidylinositol 5-phosphate, phosphatidylinositol 4,5-bisphosphate, membrane microdomains (rafts) and phosphatidylinositol 3,4,5-trisphosphate in regulation of membrane fusion (acrosome reaction, sperm-egg fusion, cortical granule exocytosis), inositol 1,4,5-trisphosphate receptors, and calcium release are discussed. The role of six lipases involved in generating putative lipid second messengers during fertilization is also discussed: phospholipase D, autotaxin, lipin1, sphingomyelinase, phospholipase C, and phospholipase A2. More specifically, proteins involved in developmental events and their regulation through lipid binding to SH3, SH4, PH, PX, or C2 protein domains is emphasized. New models are presented for PA activation of Src (through SH3, SH4 and a unique domain), that this may be why the SH2 domain of PLCγ is not required for Xenopus fertilization, PA activation of phospholipase C, a role for PA during the calcium wave after fertilization, and that calcium/calmodulin may be responsible for the loss of Src from rafts after fertilization. Also discussed is that the large DAG increase during fertilization derives from phospholipase D production of PA and lipin dephosphorylation to DAG. PMID:25748412

  18. Rapid phosphatidic acid accumulation in response to low temperature stress in Arabidopsis is generated through diacylglycerol kinase.

    PubMed

    Arisz, Steven A; van Wijk, Ringo; Roels, Wendy; Zhu, Jian-Kang; Haring, Michel A; Munnik, Teun

    2013-01-01

    Phosphatidic acid (PtdOH) is emerging as an important signaling lipid in abiotic stress responses in plants. The effect of cold stress was monitored using (32)P-labeled seedlings and leaf discs of Arabidopsis thaliana. Low, non-freezing temperatures were found to trigger a very rapid (32)P-PtdOH increase, peaking within 2 and 5 min, respectively. In principle, PtdOH can be generated through three different pathways, i.e., (1) via de novo phospholipid biosynthesis (through acylation of lyso-PtdOH), (2) via phospholipase D hydrolysis of structural phospholipids, or (3) via phosphorylation of diacylglycerol (DAG) by DAG kinase (DGK). Using a differential (32)P-labeling protocol and a PLD-transphosphatidylation assay, evidence is provided that the rapid (32)P-PtdOH response was primarily generated through DGK. A simultaneous decrease in the levels of (32)P-PtdInsP, correlating in time, temperature dependency, and magnitude with the increase in (32)P-PtdOH, suggested that a PtdInsP-hydrolyzing PLC generated the DAG in this reaction. Testing T-DNA insertion lines available for the seven DGK genes, revealed no clear changes in (32)P-PtdOH responses, suggesting functional redundancy. Similarly, known cold-stress mutants were analyzed to investigate whether the PtdOH response acted downstream of the respective gene products. The hos1, los1, and fry1 mutants were found to exhibit normal PtdOH responses. Slight changes were found for ice1, snow1, and the overexpression line Super-ICE1, however, this was not cold-specific and likely due to pleiotropic effects. A tentative model illustrating direct cold effects on phospholipid metabolism is presented. PMID:23346092

  19. Membrane metabolism mediated by Sec14 family members influences Arf GTPase activating protein activity for transport from the trans-Golgi.

    PubMed

    Wong, Tania A; Fairn, Gregory D; Poon, Pak P; Shmulevitz, Maya; McMaster, Christopher R; Singer, Richard A; Johnston, Gerald C

    2005-09-01

    The budding yeast Saccharomyces cerevisiae contains a family of Arf (ADP-ribosylation factor) GTPase activating protein (GAP) proteins with the Gcs1 + Age2 ArfGAP pair providing essential overlapping function for the movement of transport vesicles from the trans-Golgi network. We have generated a temperature-sensitive but stable version of the Gcs1 protein that is impaired only for trans-Golgi transport and find that deleterious effects of this enfeebled Gcs1-4 mutant protein are relieved by increased gene dosage of the gcs1-4 mutant gene itself or by the SFH2 gene (also called CSR1), encoding a phosphatidylinositol transfer protein (PITP). This effect was not seen for the SEC14 gene, encoding the founding member of the yeast PITP protein family, even though the Gcs1 and Age2 ArfGAPs are known to be downstream effectors of Sec14-mediated activity for trans-Golgi transport. Sfh2-mediated suppression of inadequate Gcs1-4 function depended on phospholipase D, whereas inadequate Gcs1-4 activity was relieved by increasing levels of diacylglycerol (DAG). Recombinant Gcs1 protein was found to bind certain phospholipids but not DAG. Our findings favor a model of Gcs1 localization through binding to specific phospholipids and activation of ArfGAP activity by DAG-mediated membrane curvature as the transport vesicle is formed. Thus, ArfGAPs are subject to both temporal and spatial regulation that is facilitated by Sfh2-mediated modulation of the lipid environment. PMID:16126894

  20. Crystallization kinetics of olivine-phyric shergottites

    NASA Astrophysics Data System (ADS)

    Ennis, Megan E.; McSween, Harry Y.

    2014-08-01

    Crystal size distribution (CSD) and spatial distribution pattern (SDP) analyses are applied to the early crystallizing phases, olivine and pyroxene, in olivine-phyric shergottites (Elephant moraine [EET] 79001A, Dar al Gani [DaG] 476, and dhofar [Dho] 019) from each sampling locality inferred from Mars ejection ages. Trace element zonation patterns (P and Cr) in olivine are also used to characterize the crystallization history of these Martian basalts. Previously reported 2-D CSDs for these meteorites are re-evaluated using a newer stereographically corrected methodology. Kinks in the olivine CSD plots suggest several populations that crystallized under different conditions. CSDs for pyroxene in DaG 476 and EET 79001A reveal single populations that grew under steady-state conditions; pyroxenes in Dho 019 were too intergrown for CSD analysis. Magma chamber residence times of several days for small grains to several months for olivine megacrysts are calculated using the CSD slopes and growth rates inferred from previous experimental data. Phosphorus imaging in olivines in DaG 476 and Dho 019 indicate rapid growth of skeletal, sector-zoned, or patchy cores, probably in response to delayed nucleation, followed by slow growth, and finally rapid dendritic growth with back-filling to form oscillatory zoning in rims. SPD analyses indicate that olivine and pyroxene crystals grew or accumulated in clusters rather than as randomly distributed grains. These data reveal complex solidification histories for Martian basalts, and are generally consistent with the formation at depth of olivine megacryst cores, which were entrained in ascending magmas that crystallized pyroxenes, small olivines, and oscillatory rims on megacrysts.

  1. Tracking synthesis and turnover of triacylglycerol in leaves

    PubMed Central

    Tjellström, Henrik; Strawsine, Merissa; Ohlrogge, John B.

    2015-01-01

    Triacylglycerol (TAG), typically represents <1% of leaf glycerolipids but can accumulate under stress and other conditions or if leaves are supplied with fatty acids, or in plants transformed with regulators or enzymes of lipid metabolism. To better understand the metabolism of TAG in leaves, pulse–chase radiolabelling experiments were designed to probe its synthesis and turnover. When Arabidopsis leaves were incubated with [14C]lauric acid (12:0), a major initial product was [14C]TAG. Thus, despite low steady-state levels, leaves possess substantial TAG biosynthetic capacity. The contributions of diacylglycerol acyltransferase1 and phospholipid:diacylglycerol acyltransferase1 to leaf TAG synthesis were examined by labelling of dgat1 and pdat1 mutants. The dgat1 mutant displayed a major (76%) reduction in [14C]TAG accumulation whereas pdat1 TAG labelling was only slightly reduced. Thus, DGAT1 has a principal role in TAG biosynthesis in young leaves. During a 4h chase period, radioactivity in TAG declined 70%, whereas the turnover of [14C]acyl chains of phosphatidylcholine (PC) and other polar lipids was much lower. Sixty percent of [14C]12:0 was directly incorporated into glycerolipids without modification, whereas 40% was elongated and desaturated to 16:0 and 18:1 by plastids. The unmodified [14C]12:0 and the plastid products of [14C]12:0 metabolism entered different pathways. Although plastid-modified 14C-labelled products accumulated in monogalactosyldiacylglycerol, PC, phosphatidylethanolamine, and diacylglcerol (DAG), there was almost no accumulation of [14C]16:0 and [14C]18:1 in TAG. Because DAG and acyl-CoA are direct precursors of TAG, the differential labelling of polar glycerolipids and TAG by [14C]12:0 and its plastid-modified products provides evidence for multiple subcellular pools of both acyl-CoA and DAG. PMID:25609824

  2. Salicylic acid modulates levels of phosphoinositide dependent-phospholipase C substrates and products to remodel the Arabidopsis suspension cell transcriptome

    PubMed Central

    Ruelland, Eric; Pokotylo, Igor; Djafi, Nabila; Cantrel, Catherine; Repellin, Anne; Zachowski, Alain

    2014-01-01

    Basal phosphoinositide-dependent phospholipase C (PI-PLC) activity controls gene expression in Arabidopsis suspension cells and seedlings. PI-PLC catalyzes the production of phosphorylated inositol and diacylglycerol (DAG) from phosphoinositides. It is not known how PI-PLC regulates the transcriptome although the action of DAG-kinase (DGK) on DAG immediately downstream from PI-PLC is responsible for some of the regulation. We previously established a list of genes whose expression is affected in the presence of PI-PLC inhibitors. Here this list of genes was used as a signature in similarity searches of curated plant hormone response transcriptome data. The strongest correlations obtained with the inhibited PI-PLC signature were with salicylic acid (SA) treatments. We confirm here that in Arabidopsis suspension cells SA treatment leads to an increase in phosphoinositides, then demonstrate that SA leads to a significant 20% decrease in phosphatidic acid, indicative of a decrease in PI-PLC products. Previous sets of microarray data were re-assessed. The SA response of one set of genes was dependent on phosphoinositides. Alterations in the levels of a second set of genes, mostly SA-repressed genes, could be related to decreases in PI-PLC products that occur in response to SA action. Together, the two groups of genes comprise at least 40% of all SA-responsive genes. Overall these two groups of genes are distinct in the functional categories of the proteins they encode, their promoter cis-elements and their regulation by DGK or phospholipase D. SA-regulated genes dependent on phosphoinositides are typical SA response genes while those with an SA response that is possibly dependent on PI-PLC products are less SA-specific. We propose a model in which SA inhibits PI-PLC activity and alters levels of PI-PLC products and substrates, thereby regulating gene expression divergently. PMID:25426125

  3. Muscarinic receptor activation of phosphatidylcholine hydrolysis. Relationship to phosphoinositide hydrolysis and diacylglycerol metabolism

    SciTech Connect

    Martinson, E.A.; Goldstein, D.; Brown, J.H. )

    1989-09-05

    We examined the relationship between phosphatidylcholine (PC) hydrolysis, phosphoinositide hydrolysis, and diacylglycerol (DAG) formation in response to muscarinic acetylcholine receptor (mAChR) stimulation in 1321N1 astrocytoma cells. Carbachol increases the release of (3H)choline and (3H)phosphorylcholine ((3H)Pchol) from cells containing (3H)choline-labeled PC. The production of Pchol is rapid and transient, while choline production continues for at least 30 min. mAChR-stimulated release of Pchol is reduced in cells that have been depleted of intracellular Ca2+ stores by ionomycin pretreatment, whereas choline release is unaffected by this pretreatment. Phorbol 12-myristate 13-acetate (PMA) increases the release of choline, but not Pchol, from 1321N1 cells, and down-regulation of protein kinase C blocks the ability of carbachol to stimulate choline production. Taken together, these results suggest that Ca2+ mobilization is involved in mAChR-mediated hydrolysis of PC by a phospholipase C, whereas protein kinase C activation is required for mAChR-stimulated hydrolysis of PC by a phospholipase D. Both carbachol and PMA rapidly increase the formation of (3H)phosphatidic acid ((3H)PA) in cells containing (3H)myristate-labeled PC. (3H)Diacylglycerol ((3H)DAG) levels increase more slowly, suggesting that the predominant pathway for PC hydrolysis is via phospholipase D. When cells are labeled with (3H)myristate and (14C)arachidonate such that there is a much greater 3H/14C ratio in PC compared with the phosphoinositides, the 3H/14C ratio in DAG and PA increases with PMA treatment but decreases in response to carbachol.

  4. PKCε plays a causal role in acute ethanol-induced steatosis

    PubMed Central

    Kaiser, J. Phillip; Beier, Juliane I.; Zhang, Jun; Hoetker, J. David; von Montfort, Claudia; Guo, Luping; Zheng, Yuting; Monia, Brett P.; Bhatnagar, Aruni; Arteel, Gavin E.

    2009-01-01

    Steatosis is a critical stage in the pathology of alcoholic liver disease (ALD), and preventing steatosis could protect against later stages of ALD. PKCε has been shown to contribute to hepatic steatosis in experimental non-alcoholic fatty liver disease (NAFLD); however, the role of PKCε in ethanol-induced steatosis has not been determined. The purpose of this study was to therefore test the hypothesis that PKCε contributes to ethanol-induced steatosis. Accordingly, the effect of acute ethanol on indices of hepatic steatosis and insulin signaling were determined in PKCε knockout mice and in wild-type mice that received an antisense oligonucleotide (ASO) to knockdown PKCε expression. Acute ethanol (6 g/kg i.g.) caused a robust increase in hepatic non-esterified free fatty acids (NEFA), which peaked 1 h after ethanol exposure. This increase in NEFA was followed by elevated diacylglycerols (DAG), as well as by the concomitant activation of PKCε. Acute ethanol also changed the expression of insulin-responsive genes (i.e. increased G6Pase, downregulated GK), in a pattern indicative of impaired insulin signaling. Acute ethanol exposure subsequently caused a robust increase in hepatic triglycerides. The accumulation of triglycerides caused by ethanol was blunted in ASO-treated or in PKCε−/− mice. Taken together, these data suggest that the increase in NEFA caused by hepatic ethanol metabolism leads to an increase in DAG production via the triacylglycerol pathway. DAG then subsequently activates PKCε, which then exacerbates hepatic lipid accumulation by inducing insulin resistance. These data also suggest that PKCε plays a causal role in at least the early phases of ethanol-induced liver injury. PMID:19022218

  5. Semi-automated ontology generation within OBO-Edit

    PubMed Central

    Wächter, Thomas; Schroeder, Michael

    2010-01-01

    Motivation: Ontologies and taxonomies have proven highly beneficial for biocuration. The Open Biomedical Ontology (OBO) Foundry alone lists over 90 ontologies mainly built with OBO-Edit. Creating and maintaining such ontologies is a labour-intensive, difficult, manual process. Automating parts of it is of great importance for the further development of ontologies and for biocuration. Results: We have developed the Dresden Ontology Generator for Directed Acyclic Graphs (DOG4DAG), a system which supports the creation and extension of OBO ontologies by semi-automatically generating terms, definitions and parent–child relations from text in PubMed, the web and PDF repositories. DOG4DAG is seamlessly integrated into OBO-Edit. It generates terms by identifying statistically significant noun phrases in text. For definitions and parent–child relations it employs pattern-based web searches. We systematically evaluate each generation step using manually validated benchmarks. The term generation leads to high-quality terms also found in manually created ontologies. Up to 78% of definitions are valid and up to 54% of child–ancestor relations can be retrieved. There is no other validated system that achieves comparable results. By combining the prediction of high-quality terms, definitions and parent–child relations with the ontology editor OBO-Edit we contribute a thoroughly validated tool for all OBO ontology engineers. Availability: DOG4DAG is available within OBO-Edit 2.1 at http://www.oboedit.org Contact: thomas.waechter@biotec.tu-dresden.de; Supplementary Information: Supplementary data are available at Bioinformatics online. PMID:20529942

  6. Lipase-catalyzed methanolysis of triricinolein in organic solvent to produce 1,2(2,3)-diricinolein.

    PubMed

    Turner, Charlotta; He, Xiaohua; Nguyen, Tasha; Lin, Jiann-Tsyh; Wong, Rosalind Y; Lundin, Robert E; Harden, Leslie; McKeon, Thomas

    2003-11-01

    The objective of this study was to find the optimal parameters for lipase-catalyzed methanolysis of triricinolein to produce 1,2(2,3)-diricinolein. Four different immobilized lipases were tested, Candida antarctica type B (CALB), Rhizomucor miehei (RML), Pseudomonas cepacia (PCL), and Penicillium roquefortii (PRL). n-Hexane and diisopropyl ether (DIPE) were examined as reaction media at three different water activities (a(w)), 0.11, 0.53, and 0.97. The consumption of triricinolein and the formation of 1,2(2,3)-diricinolein, methyl ricinoleate, and ricinoleic acid were followed for up to 48 h. PRL gave the highest yield of 1,2(2,3)-diricinolein. Moreover, this lipase showed the highest specificity for the studied reaction, i.e., high selectivity for the reaction with triricinolein but low for 1,2(2,3)-diricinolein. Recoveries of 93 and 88% DAG were obtained using PRL in DIPE at a(w) of 0.11 and 0.53, respectively. Further, NMR studies showed that a higher purity of the 1,2(2,3)-isomer vs. the 1,3-isomer was achieved at higher a(w) (88% at a(w) = 0.53), compared to lower a(w) (71% at a(w) = 0.11). The DAG obtained was acylated by the DAG acyltransferase from Arabidopsis thaliana. Therefore, this enzymatic product is a useful enzyme substrate for lipid biosynthesis. Accordingly, the use of PRL in DIPE at a(w) 0.53 is considered optimal for the synthesis of 1,2(2,3)-diricinolein from triricinolein. PMID:14733366

  7. PC-PLC/sphingomyelin synthase activity plays a central role in the development of myogenic tone in murine resistance arteries

    PubMed Central

    Zacharia, Joseph; Fairfax, Seth; Wier, Withrow Gil

    2015-01-01

    Myogenic tone is an intrinsic property of the vasculature that contributes to blood pressure control and tissue perfusion. Earlier investigations assigned a key role in myogenic tone to phospholipase C (PLC) and its products, inositol 1,4,5-trisphosphate (IP3) and diacylglycerol (DAG). Here, we used the PLC inhibitor, U-73122, and two other, specific inhibitors of PLC subtypes (PI-PLC and PC-PLC) to delineate the role of PLC in myogenic tone of pressurized murine mesenteric arteries. U-73122 inhibited depolarization-induced contractions (high external K+ concentration), thus confirming reports of nonspecific actions of U-73122 and its limited utility for studies of myogenic tone. Edelfosine, a specific inhibitor of PI-PLC, did not affect depolarization-induced contractions but modulated myogenic tone. Because PI-PLC produces IP3, we investigated the effect of blocking IP3 receptor-mediated Ca2+ release on myogenic tone. Incubation of arteries with xestospongin C did not affect tone, consistent with the virtual absence of Ca2+ waves in arteries with myogenic tone. D-609, an inhibitor of PC-PLC and sphingomyelin synthase, strongly inhibited myogenic tone and had no effect on depolarization-induced contraction. D-609 appeared to act by lowering cytoplasmic Ca2+ concentration to levels below those that activate contraction. Importantly, incubation of pressurized arteries with a membrane-permeable analog of DAG induced vasoconstriction. The results therefore mandate a reexamination of the signaling pathways activated by the Bayliss mechanism. Our results suggest that PI-PLC and IP3 are not required in maintaining myogenic tone, but DAG, produced by PC-PLC and/or SM synthase, is likely through multiple mechanisms to increase Ca2+ entry and promote vasoconstriction. PMID:25888510

  8. Troglitazone stimulates {beta}-arrestin-dependent cardiomyocyte contractility via the angiotensin II type 1{sub A} receptor

    SciTech Connect

    Tilley, Douglas G.; Nguyen, Anny D.; Rockman, Howard A.; Department of Cell Biology, Duke University Medical Center; Department of Molecular Genetics and Microbiology, Duke University Medical Center

    2010-06-11

    Peroxisome proliferator-activated receptor {gamma} (PPAR{gamma}) agonists are commonly used to treat cardiovascular diseases, and are reported to have several effects on cardiovascular function that may be due to PPAR{gamma}-independent signaling events. Select angiotensin receptor blockers (ARBs) interact with and modulate PPAR{gamma} activity, thus we hypothesized that a PPAR{gamma} agonist may exert physiologic effects via the angiotensin II type 1{sub A} receptor (AT1{sub A}R). In AT1{sub A}R-overexpressing HEK 293 cells, both angiotensin II (Ang II) and the PPAR{gamma} agonist troglitazone (Trog) enhanced AT1{sub A}R internalization and recruitment of endogenous {beta}-arrestin1/2 ({beta}arr1/2) to the AT1{sub A}R. A fluorescence assay to measure diacylglycerol (DAG) accumulation showed that although Ang II induced AT1{sub A}R-G{sub q} protein-mediated DAG accumulation, Trog had no impact on DAG generation. Trog-mediated recruitment of {beta}arr1/2 was selective to AT1{sub A}R as the response was prevented by an ARB- and Trog-mediated {beta}arr1/2 recruitment to {beta}1-adrenergic receptor ({beta}1AR) was not observed. In isolated mouse cardiomyocytes, Trog increased both % and rate of cell shortening to a similar extent as Ang II, effects which were blocked with an ARB. Additionally, these effects were found to be {beta}arr2-dependent, as cardiomyocytes isolated from {beta}arr2-KO mice showed blunted contractile responses to Trog. These findings show for the first time that the PPAR{gamma} agonist Trog acts at the AT1{sub A}R to simultaneously block G{sub q} protein activation and induce the recruitment of {beta}arr1/2, which leads to an increase in cardiomyocyte contractility.

  9. Platelet activation by bacterial phospholipase C involves phosphoinositide turnover and phosphorylation of 47,000 dalton but not 20,000 dalton protein

    SciTech Connect

    Huzoor-Akbar; Anwer, K.

    1986-05-01

    This study was conducted to examine the role of phosphoinositides (PIns) and phosphorylation of 47,000 dalton (P47) and 20,000 dalton (P20) proteins in platelet activation by bacterial phospholipase C (PLC). PLC induced serotonin secretion (SS) and platelet aggregation (PA) in a concentration dependent manner. PLC (0.02 U/ml) caused phosphorylation of P47 in a time dependent manner (27% at 0.5 min to 378% at 7 min). PLC did not induce more than 15% phosphorylation of P20 by 7 min. Aspirin (500 ..mu..M) blocked phosphorylation of P20 but did not inhibit SS, PA or phosphorylation of P47. PLC (0.04 U/ml) decreased radioactivity (cpm) in /sup 32/P labeled phosphatidylinositol (PI), PI-4,5-bis-PO4 (PIP2) and PI-4-PO4 (PIP) by 20%, 12% and 7.5% respectively at 15 sec. The level of PI but not that of PIP2 returned to base line in 3 min. PIP level increased above control values within one min. PLC increased phosphatidic acid level (75% at 0.5 min. to 1545% at 3 min). In other experiments PLC produced diacylglycerol (DAG) in a time and concentration dependent manner. However, no DAG was detectable in the first 60 sec. These data suggest that: (a) PIns turnover and phosphorylation of P47 but not that of P20 is involved in platelet activation by PLC; and (b) DAG production from outer membrane phospholipids is not a prerequisite for platelet activation by PLC.

  10. Rapid phosphatidic acid accumulation in response to low temperature stress in Arabidopsis is generated through diacylglycerol kinase

    PubMed Central

    Arisz, Steven A.; van Wijk, Ringo; Roels, Wendy; Zhu, Jian-Kang; Haring, Michel A.; Munnik, Teun

    2013-01-01

    Phosphatidic acid (PtdOH) is emerging as an important signaling lipid in abiotic stress responses in plants. The effect of cold stress was monitored using 32P-labeled seedlings and leaf discs of Arabidopsis thaliana. Low, non-freezing temperatures were found to trigger a very rapid 32P-PtdOH increase, peaking within 2 and 5 min, respectively. In principle, PtdOH can be generated through three different pathways, i.e., (1) via de novo phospholipid biosynthesis (through acylation of lyso-PtdOH), (2) via phospholipase D hydrolysis of structural phospholipids, or (3) via phosphorylation of diacylglycerol (DAG) by DAG kinase (DGK). Using a differential 32P-labeling protocol and a PLD-transphosphatidylation assay, evidence is provided that the rapid 32P-PtdOH response was primarily generated through DGK. A simultaneous decrease in the levels of 32P-PtdInsP, correlating in time, temperature dependency, and magnitude with the increase in 32P-PtdOH, suggested that a PtdInsP-hydrolyzing PLC generated the DAG in this reaction. Testing T-DNA insertion lines available for the seven DGK genes, revealed no clear changes in 32P-PtdOH responses, suggesting functional redundancy. Similarly, known cold-stress mutants were analyzed to investigate whether the PtdOH response acted downstream of the respective gene products. The hos1, los1, and fry1 mutants were found to exhibit normal PtdOH responses. Slight changes were found for ice1, snow1, and the overexpression line Super-ICE1, however, this was not cold-specific and likely due to pleiotropic effects. A tentative model illustrating direct cold effects on phospholipid metabolism is presented. PMID:23346092

  11. A mutation in TRPC6 channels abolishes their activation by hypoosmotic stretch but does not affect activation by diacylglycerol or G protein signaling cascades.

    PubMed

    Wilson, Cory; Dryer, Stuart E

    2014-05-01

    Canonical transient receptor potential-6 (TRPC6) channels have been implicated in the pathogenesis of kidney disease and in the regulation of vascular smooth muscle tone, podocyte function, and a variety of processes in other cell types. The question of whether their gating is intrinsically mechanosensitive has been controversial. In this study we have examined activation of two alleles of TRPC6 transiently expressed in CHO-K1 cells: the wild-type human TRPC6 channel, and TRPC6-N143S, an allele originally identified in a family with autosomal dominant familial focal and segmental glomerulosclerosis (FSGS). We observed that both channel variants carried robust cationic currents that could be evoked by application of membrane-permeable analogs of diacylglycerol (DAG) or by the P2Y receptor agonist ATP. The amplitudes and characteristics of currents evoked by the DAG analog or ATP were indistinguishable in cells expressing the two TRPC6 alleles. By contrast, hypoosmotic stretch evoked robust currents in wild-type TRPC6 channels but had no discernible effect on currents in cells expressing TRPC6-N143S, indicating that the mutant form lacks mechanosensitivity. Coexpression of TRPC6-N143S with wild-type TRPC6 or TRPC3 channels did not alter stretch-evoked responses compared with when TRPC3 channels were expressed by themselves, indicating that TRPC6-N143S does not function as a dominant-negative. These data indicate that mechanical activation and activation evoked by DAG or ATP occur through fundamentally distinct biophysical mechanisms, and they provide support for the hypothesis that protein complexes containing wild-type TRPC6 subunits can be intrinsically mechanosensitive. PMID:24598806

  12. High-fat diet-mediated lipotoxicity and insulin resistance is related to impaired lipase expression in mouse skeletal muscle.

    PubMed

    Badin, Pierre-Marie; Vila, Isabelle K; Louche, Katie; Mairal, Aline; Marques, Marie-Adeline; Bourlier, Virginie; Tavernier, Geneviève; Langin, Dominique; Moro, Cedric

    2013-04-01

    Elevated expression/activity of adipose triglyceride lipase (ATGL) and/or reduced activity of hormone-sensitive lipase (HSL) in skeletal muscle are causally linked to insulin resistance in vitro. We investigated here the effect of high-fat feeding on skeletal muscle lipolytic proteins, lipotoxicity, and insulin signaling in vivo. Five-week-old C3H mice were fed normal chow diet (NCD) or 45% kcal high-fat diet (HFD) for 4 weeks. Wild-type and HSL knockout mice fed NCD were also studied. Whole-body and muscle insulin sensitivity, as well as lipolytic protein expression, lipid levels, and insulin signaling in skeletal muscle, were measured. HFD induced whole-body insulin resistance and glucose intolerance and reduced skeletal muscle glucose uptake compared with NCD. HFD increased skeletal muscle total diacylglycerol (DAG) content, protein kinase Cθ and protein kinase Cε membrane translocation, and impaired insulin signaling as reflected by a robust increase of basal Ser1101 insulin receptor substrate 1 phosphorylation (2.8-fold, P < .05) and a decrease of insulin-stimulated v-Akt murine thymoma viral oncogene homolog Ser473 (-37%, P < .05) and AS160 Thr642 (-47%, P <.01) phosphorylation. We next showed that HFD strongly reduced HSL phosphorylation at Ser660. HFD significantly up-regulated the muscle protein content of the ATGL coactivator comparative gene identification 58 and triacylglycerol hydrolase activity, despite a lower ATGL protein content. We further show a defective skeletal muscle insulin signaling and DAG accumulation in HSL knockout compared with wild-type mice. Together, these data suggest a pathophysiological link between altered skeletal muscle lipase expression and DAG-mediated insulin resistance in mice. PMID:23471217

  13. Altered Skeletal Muscle Fatty Acid Handling in Subjects with Impaired Glucose Tolerance as Compared to Impaired Fasting Glucose.

    PubMed

    Goossens, Gijs H; Moors, Chantalle C M; Jocken, Johan W E; van der Zijl, Nynke J; Jans, Anneke; Konings, Ellen; Diamant, Michaela; Blaak, Ellen E

    2016-03-01

    Altered skeletal muscle fatty acid (FA) metabolism contributes to insulin resistance. Here, we compared skeletal muscle FA handling between subjects with impaired fasting glucose (IFG; n = 12 (7 males)) and impaired glucose tolerance (IGT; n = 14 (7 males)) by measuring arterio-venous concentration differences across forearm muscle. [²H₂]-palmitate was infused intravenously, labeling circulating endogenous triacylglycerol (TAG) and free fatty acids (FFA), whereas [U-(13)C]-palmitate was incorporated in a high-fat mixed-meal, labeling chylomicron-TAG. Skeletal muscle biopsies were taken to determine muscle TAG, diacylglycerol (DAG), FFA, and phospholipid content, their fractional synthetic rate (FSR) and degree of saturation, and gene expression. Insulin sensitivity was assessed using a hyperinsulinemic-euglycemic clamp. Net skeletal muscle glucose uptake was lower (p = 0.018) and peripheral insulin sensitivity tended to be reduced (p = 0.064) in IGT as compared to IFG subjects. Furthermore, IGT showed higher skeletal muscle extraction of VLDL-TAG (p = 0.043), higher muscle TAG content (p = 0.025), higher saturation of FFA (p = 0.004), lower saturation of TAG (p = 0.017) and a tendency towards a lower TAG FSR (p = 0.073) and a lower saturation of DAG (p = 0.059) versus IFG individuals. Muscle oxidative gene expression was lower in IGT subjects. In conclusion, increased liver-derived TAG extraction and reduced lipid turnover of saturated FA, rather than DAG content, in skeletal muscle accompany the more pronounced insulin resistance in IGT versus IFG subjects. PMID:26985905

  14. Metabolic labelling of membrane microdomains/rafts in Jurkat cells indicates the presence of glycerophospholipids implicated in signal transduction by the CD3 T-cell receptor.

    PubMed Central

    Rouquette-Jazdanian, Alexandre K; Pelassy, Claudette; Breittmayer, Jean-Philippe; Cousin, Jean-Louis; Aussel, Claude

    2002-01-01

    Cell membranes contain sphingolipids and cholesterol, which cluster together in distinct domains called rafts. The outer-membrane leaflet of these peculiar membrane domains contains glycosylphosphatidylinositol-anchored proteins, while the inner leaflet contains proteins implicated in signalling, such as the acylated protein kinase p56(lck) and the palmitoylated adaptator LAT (linker for activation of T-cells). We present here an approach to study the lipid composition of rafts and its change upon T-cell activation. Our method is based on metabolic labelling of Jurkat T-cells with different precursors of glycerophospholipid synthesis, including glycerol and fatty acids with different lengths and degrees of saturation as well as phospholipid polar head groups. The results obtained indicate that lipid rafts isolated by the use of sucrose density-gradient centrifugation after Triton X-100 extraction in the cold, besides sphingolipids and cholesterol, contain unambiguously all classes of glycerophospholipids: phosphatidylserine, phosphatidylinositol, phosphatidylethanolamine and phosphatidylcholine. Fatty acid labelling shows that lipid rafts are labelled preferentially with saturated fatty acids while the rest of the plasma membrane incorporates mostly long-chained polyunsaturated fatty acids. To see whether the raft composition as measured by metabolic labelling of phospholipids is involved in T-cell activation, we investigated the production of sn-1,2-diacylglycerol (DAG) in CD3-activated cells. DAG production occurs within rafts, confirming previous demonstration of protein kinase C translocation into membrane microdomains. Our data demonstrate that raft disorganization by methyl-beta-cyclodextrin impairs both CD3-induced DAG production and changes in cytosolic Ca(2+) concentration. These lines of evidence support the conclusion that the major events in T-cell activation occur within or due to lipid rafts. PMID:11964165

  15. Altered lipid composition in Streptococcus pneumoniae cpoA mutants

    PubMed Central

    2014-01-01

    Background Penicillin-resistance in Streptococcus pneumoniae is mainly due to alterations in genes encoding the target enzymes for beta-lactams, the penicillin-binding proteins (PBPs). However, non-PBP genes are altered in beta-lactam-resistant laboratory mutants and confer decreased susceptibility to beta-lactam antibiotics. Two piperacillin resistant laboratory mutants of Streptococcus pneumoniae R6 contain mutations in the putative glycosyltransferase gene cpoA. The CpoA gene is part of an operon including another putative glycosyltransferase gene spr0982, both of which being homologous to glycolipid synthases present in other Gram-positive bacteria. Results We now show that the cpoA mutants as well as a cpoA deletion mutant are defective in the synthesis of galactosyl-glucosyl-diacylglycerol (GalGlcDAG) in vivo consistent with the in vitro function of CpoA as α-GalGlcDAG synthase as shown previously. In addition, the proportion of phosphatidylglycerol increased relative to cardiolipin in cpoA mutants. Moreover, cpoA mutants are more susceptible to acidic stress, have an increased requirement for Mg2+ at low pH, reveal a higher resistance to lysis inducing conditions and are hypersensitive to bacitracin. Conclusions The data show that deficiency of the major glycolipid GalGlcDAG causes a pleitotropic phenotype of cpoA mutant cells consistent with severe membrane alterations. We suggest that the cpoA mutations selected with piperacillin are directed against the lytic response induced by the beta-lactam antibiotic. PMID:24443834

  16. Identification of Two Legionella pneumophila Effectors that Manipulate Host Phospholipids Biosynthesis

    PubMed Central

    Viner, Ram; Chetrit, David; Ehrlich, Marcelo; Segal, Gil

    2012-01-01

    The intracellular pathogen Legionella pneumophila translocates a large number of effector proteins into host cells via the Icm/Dot type-IVB secretion system. Some of these effectors were shown to cause lethal effect on yeast growth. Here we characterized one such effector (LecE) and identified yeast suppressors that reduced its lethal effect. The LecE lethal effect was found to be suppressed by the over expression of the yeast protein Dgk1 a diacylglycerol (DAG) kinase enzyme and by a deletion of the gene encoding for Pah1 a phosphatidic acid (PA) phosphatase that counteracts the activity of Dgk1. Genetic analysis using yeast deletion mutants, strains expressing relevant yeast genes and point mutations constructed in the Dgk1 and Pah1 conserved domains indicated that LecE functions similarly to the Nem1-Spo7 phosphatase complex that activates Pah1 in yeast. In addition, by using relevant yeast genetic backgrounds we examined several L. pneumophila effectors expected to be involved in phospholipids biosynthesis and identified an effector (LpdA) that contains a phospholipase-D (PLD) domain which caused lethal effect only in a dgk1 deletion mutant of yeast. Additionally, LpdA was found to enhance the lethal effect of LecE in yeast cells, a phenomenon which was found to be dependent on its PLD activity. Furthermore, to determine whether LecE and LpdA affect the levels or distribution of DAG and PA in-vivo in mammalian cells, we utilized fluorescent DAG and PA biosensors and validated the notion that LecE and LpdA affect the in-vivo levels and distribution of DAG and PA, respectively. Finally, we examined the intracellular localization of both LecE and LpdA in human macrophages during L. pneumophila infection and found that both effectors are localized to the bacterial phagosome. Our results suggest that L. pneumophila utilize at least two effectors to manipulate important steps in phospholipids biosynthesis. PMID:23133385

  17. Some Lithuanian ethnobotanical taxa: a linguistic view on Thorn Apple and related plants

    PubMed Central

    Šeškauskaitė, Daiva; Gliwa, Bernd

    2006-01-01

    Background The perception and use of plants correspond with common plant names. The study of plant names may give insight into historical and recent use of plants. Methods Plant names in dictionaries and folklore have been evaluated. A etymological analysis of the names is provided. Onomasiological and semasiological aspects have been considered. Therefore, species named with names related to each other have been selected. Results Plant names containing the stem dag- or deg- may belong to either of two categories: incenses or thorny plants. Plants named in durn- have been in use as psychopharmaca. The name rymo points not to Rome but to the use of plants as anodyne or psychopharmaca. PMID:16512899

  18. Water-extractable and exchangeable phosphate in Martian and carbonaceous chondrite meteorites and in planetary soil analogs

    NASA Astrophysics Data System (ADS)

    Mautner, Michael N.; Sinaj, Sokrat

    2002-09-01

    Aqueous extraction contributes to the formation and weathering of planetary materials and renders electrolytes such as phosphate available for biology. In this context, the solubility of phosphate is measured in planetary materials, represented by the Mars meteorites Nakhla, Dar al Gani 476 (DaG 476), Elephant Morraine 79001 (EETA 79001), and terrestrial analogs, and in the Murchison CM2 and Allende CV3 carbonaceous chondrites. The Mars meteorites contain high levels of phosphate that is readily extracted by water, up to 15 mg kg -1 in Nakhla and DaG 476 and 38 mg kg -1 in EETA 79001, while the terrestrial analogs and the carbonaceous chondrites contain 0.5 to 6 mg kg -1. Correspondingly, high phosphate concentrations of 4 to >28 mg L -1 are obtained in extracts of the Mars meteorites at high solid/solution ratios, exceeding the concentrations of 0.4 to 2.0 mg L -1 in the extracts of the terrestrial analogs. A wide range of planetary conditions, including N 2 and CO 2 atmospheres, solid/solution ratios of 0.01 to 1.0 kg L -1, extraction times of 1 to 21 d, and temperatures of 20 to 121°C affect the amounts of extractable phosphate by factors of only 2 to 5 in most materials. Phosphate-fixing capacity and exchangeable phosphate are assessed by the isotopic exchange kinetics (IEK) method, which quantifies the amount of P isotopically exchangeable within 1 min (E 1min) and between 1 min and 3 months (E 1min-3m) and the amount of P that cannot be exchanged within 3 months (E >3m). The IEK results show that the DaG 476 Mars meteorite and terrestrial analogs have low P-fixing capacities, while the carbonaceous chondrites have high P-fixing capacities. Aqueous processing under early planetary CO 2 atmospheres has large effects on the available phosphate. For example, the fraction of total P that is exchangeable in 3 months increases from 1.6 to 11%, 13 to 51.6%, and 43.9 to 90.4% in the DaG 476 Mars meteorite, Allende, and Murchison, respectively. The results show that

  19. Complete genome sequence of yam chlorotic necrotic mosaic virus from Dioscorea parviflora.

    PubMed

    Zhang, Pengyuan; Peng, Jiejun; Guo, Huachun; Chen, Jianping; Chen, Suiyun; Wang, Jianguang

    2016-06-01

    The complete genome sequence of yam chlorotic necrotic mosaic virus (YCNMV) was determined. It is a monopartite ssRNA 8208 nucleotides in length (excluding the poly(A) tail) and encoding a polyprotein of 2622 amino acids. Sequence analysis showed that the P1 region and some conserved motifs, such as the typical potyvirus aphid-transmission motifs DAG, PTK and KITC, are absent. Phylogenetic analysis based on the complete polyprotein sequences of YCNMV and selected members of the family Potyviridae clearly showed that this virus should be assigned to the genus Macluravirus and suggest that YCNMV is a new member of the genus Macluravirus. PMID:26973231

  20. Cutinase and hydrophobin interplay

    PubMed Central

    Gurr, Sarah J

    2008-01-01

    Surface-penetrating phytopathogenic fungi frequently form appressoria. These are specialised infection structures pivotal to fungal ingress into the host. Recently, we demonstrated that one member of a family of cutinases in Magnaporthe grisea is involved in surface sensing, mediating appressorium differentiation and penetration peg formation and hence facilitates host penetration. Cutinase2 serves as an upstream activator of cAMP/PKA and DAG/PKC signalling cascades and is essential for full virulence. Here, we speculate on the role of rice blast hydrophobins as surface interactors facilitating fungal cutinase activity. PMID:19704644

  1. An extended anyon Fock space and noncommutative Meixner-type orthogonal polynomials in infinite dimensions

    NASA Astrophysics Data System (ADS)

    Bożejko, M.; Lytvynov, E. W.; Rodionova, I. V.

    2015-10-01

    Let ν be a finite measure on R whose Laplace transform is analytic in a neighbourhood of zero. An anyon Lévy white noise on ( R^d,dx) is a certain family of noncommuting operators <ω,\\varphi> on the anyon Fock space over L^2( R^d× R,dx\\otimesν), where \\varphi=\\varphi(x) runs over a space of test functions on R^d, while ω=ω(x) is interpreted as an operator-valued distribution on R^d. Let L^2(τ) be the noncommutative L^2-space generated by the algebra of polynomials in the variables <ω,\\varphi>, where τ is the vacuum expectation state. Noncommutative orthogonal polynomials in L^2(τ) of the form < P_n(ω),f(n)> are constructed, where f(n) is a test function on ( R^d)^n, and are then used to derive a unitary isomorphism U between L^2(τ) and an extended anyon Fock space \\mathbf F(L^2( R^d,dx)) over L^2( R^d,dx). The usual anyon Fock space \\mathscr F(L^2( R^d,dx)) over L^2( R^d,dx) is a subspace of \\mathbf F(L^2( R^d,dx)). Furthermore, the equality \\mathbf F(L^2( R^d,dx))=\\mathscr F(L^2( R^d,dx)) holds if and only if the measure ν is concentrated at a single point, that is, in the Gaussian or Poisson case. With use of the unitary isomorphism U, the operators <ω,\\varphi> are realized as a Jacobi (that is, tridiagonal) field in \\mathbf F(L^2( R^d,dx)). A Meixner-type class of anyon Lévy white noise is derived for which the corresponding Jacobi field in \\mathbf F(L^2( R^d,dx)) has a relatively simple structure. Each anyon Lévy white noise of Meixner type is characterized by two parameters, λ\\in R and η≥slant0. In conclusion, the representation ω(x)=\\partial_x^\\dag +λ \\partial_x^\\dag\\partialx +η\\partial_x^\\dag\\partial_x\\partial_x+\\partial_x is obtained, where \\partial_x and \\partial_x^\\dag are the annihilation and creation operators at the point x. Bibliography: 57 titles.

  2. Genetic mapping of the mouse neuromuscular mutation kyphoscoliosis

    SciTech Connect

    Skynner, M.J.; Coulton, G.R.; Mason, R.M.

    1995-01-01

    The ky mouse mutant, kyphoscoliosis, exhibits a degenerative muscle disease resulting in chronic deformation of the spinal column. Using an interspecific backcross segregating the ky mutation, we have mapped the ky locus to a small region of mouse chromosome 9. ky is nonrecombinant with the microsatellites D9Mit24 and D9Mit169 and lies in a conserved linkage group that encompasses human chromosome 3. s-Laminin (LAMS) and the gene for dystrophin-associated glycoprotein 1 (DAG1), which map to human chromosome 3, are both recombinant with ky, ruling them out as candidates. 24 refs., 2 figs., 1 tab.

  3. Pilot Preference, Compliance, and Performance With an Airborne Conflict Management Toolset

    NASA Technical Reports Server (NTRS)

    Doble, Nathan A.; Barhydt, Richard; Krishnamurthy, Karthik

    2005-01-01

    A human-in-the-loop experiment was conducted at the NASA Ames and Langley Research Centers, investigating the En Route Free Maneuvering component of a future air traffic management concept termed Distributed Air/Ground Traffic Management (DAG-TM). NASA Langley test subject pilots used the Autonomous Operations Planner (AOP) airborne toolset to detect and resolve traffic conflicts, interacting with subject pilots and air traffic controllers at NASA Ames. Experimental results are presented, focusing on conflict resolution maneuver choices, AOP resolution guidance acceptability, and performance metrics. Based on these results, suggestions are made to further improve the AOP interface and functionality.

  4. A processor-time-minimal systolic array for cubical mesh algorithms

    SciTech Connect

    Cappello, P. . Dept. of Computer Science)

    1992-01-01

    Using a directed acyclic graph (dag) model of algorithms, the paper focuses on time-minimal multiprocessor schedules that use as few processors as possible. Such a processor-time-minimal scheduling of an algorithm's dag first is illustrated using a triangular shaped 2-D directed mesh (representing, for example, an algorithm for solving a triangular system of linear equations). Then, algorithms represented by an n {times} n {times} n directed mesh are investigated. This cubical directed mesh is fundamental; it represents the standard algorithm for computing matrix product as well as many other algorithms. Completion of the cubical mesh requires 3n - 2 steps. It is shown that the number of processing elements needed to achieve this time bound is at least (3n{sup 2/4}). A systolic array for the cubical directed mesh is then presented. It completes the mesh using the minimum number of steps and exactly (3n{sup 2/4}) processing elements: it is processor-time-minimal. The systolic array's topology is that of a hexagonally shaped, cylindrically- connected, 2-D directed mesh.

  5. Lipid-Induced Insulin Resistance in Skeletal Muscle: The Chase for the Culprit Goes from Total Intramuscular Fat to Lipid Intermediates, and Finally to Species of Lipid Intermediates

    PubMed Central

    Kitessa, Soressa M.; Abeywardena, Mahinda Y.

    2016-01-01

    The skeletal muscle is the largest organ in the body. It plays a particularly pivotal role in glucose homeostasis, as it can account for up to 40% of the body and for up to 80%–90% of insulin-stimulated glucose disposal. Hence, insulin resistance (IR) in skeletal muscle has been a focus of much research and review. The fact that skeletal muscle IR precedes β-cell dysfunction makes it an ideal target for countering the diabetes epidemic. It is generally accepted that the accumulation of lipids in the skeletal muscle, due to dietary lipid oversupply, is closely linked with IR. Our understanding of this link between intramyocellular lipids (IMCL) and glycemic control has changed over the years. Initially, skeletal muscle IR was related to total IMCL. The inconsistencies in this explanation led to the discovery that particular lipid intermediates are more important than total IMCL. The two most commonly cited lipid intermediates for causing skeletal muscle IR are ceramides and diacylglycerol (DAG) in IMCL. Still, not all cases of IR and dysfunction in glycemic control have shown an increase in either or both of these lipids. In this review, we will summarise the latest research results that, using the lipidomics approach, have elucidated DAG and ceramide species that are involved in skeletal muscle IR in animal models and human subjects. PMID:27483311

  6. Boron abundances and isotopic ratios of olivine grains on Itokawa returned by the Hayabusa spacecraft

    NASA Astrophysics Data System (ADS)

    Fujiya, Wataru; Hoppe, Peter; Ott, Ulrich

    2016-07-01

    We report the B abundances and isotopic ratios of two olivine grains from the S-type asteroid Itokawa sampled by the Hayabusa spacecraft. Olivine grains from the Dar al Gani (DaG) 989 LL6 chondrite were used as a reference. Since we analyzed polished thin sections in both cases, we expect the contribution from the solar wind B (rich in 10B) to be minimal because the solar wind was implanted only within very thin layers of the grain surface. The Itokawa and DaG 989 olivine grains have homogeneous B abundances (~400 ppb) and 11B/10B ratios compatible with the terrestrial standard and bulk chondrites. The observed homogeneous B abundances and isotopic ratios of the Itokawa olivine grains are likely the result of thermal metamorphism which occurred in the parent asteroid of Itokawa, which had a similar composition as LL chondrites. The chondritic B isotopic ratios of the Itokawa samples suggest that they contain little cosmogenic B (from cosmic-ray spallation reactions) rich in 10B. This observation is consistent with the short cosmic-ray exposure ages of Itokawa samples inferred from the small concentrations of cosmogenic 21Ne. If other Itokawa samples have little cosmogenic B as well, the enrichment in 10B found previously on the surface of another Itokawa particle (as opposed to the bulk grain study here) may be attributed to implanted solar wind B.

  7. Activation of TRPC6 channels is essential for lung ischaemia–reperfusion induced oedema in mice

    PubMed Central

    Weissmann, Norbert; Sydykov, Akylbek; Kalwa, Hermann; Storch, Ursula; Fuchs, Beate; Schnitzler, Michael Mederos y; Brandes, Ralf P.; Grimminger, Friedrich; Meissner, Marcel; Freichel, Marc; Offermanns, Stefan; Veit, Florian; Pak, Oleg; Krause, Karl-Heinz; Schermuly, Ralph T.; Brewer, Alison C; Schmidt, Harald H.H.W.; Seeger, Werner; Shah, Ajay M.; Gudermann, Thomas; Ghofrani, Hossein A.; Dietrich, Alexander

    2012-01-01

    Lung ischaemia–reperfusion-induced oedema (LIRE) is a life-threatening condition that causes pulmonary oedema induced by endothelial dysfunction. Here we show that lungs from mice lacking nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (Nox2y/−) or the classical transient receptor potential channel 6 (TRPC6−/−) are protected from LIR-induced oedema (LIRE). Generation of chimeric mice by bone marrow cell transplantation and endothelial-specific Nox2 deletion showed that endothelial Nox2, but not leukocytic Nox2 or TRPC6, are responsible for LIRE. Lung endothelial cells from Nox2- or TRPC6-deficient mice showed attenuated ischaemia-induced Ca2+ influx, cellular shape changes and impaired barrier function. Production of reactive oxygen species was completely abolished in Nox2y/− cells. A novel mechanistic model comprising endothelial Nox2-derived production of superoxide, activation of phospholipase C-γ, inhibition of diacylglycerol (DAG) kinase, DAG-mediated activation of TRPC6 and ensuing LIRE is supported by pharmacological and molecular evidence. This mechanism highlights novel pharmacological targets for the treatment of LIRE. PMID:22337127

  8. tRNA-dependent alanylation of diacylglycerol and phosphatidylglycerol in Corynebacterium glutamicum.

    PubMed

    Smith, Angela M; Harrison, Jesse S; Grube, Christopher D; Sheppe, Austin E F; Sahara, Nahoko; Ishii, Ryohei; Nureki, Osamu; Roy, Hervé

    2015-11-01

    Aminoacyl-phosphatidylglycerol synthases (aaPGSs) are membrane proteins that utilize aminoacylated tRNAs to modify membrane lipids with amino acids. Aminoacylation of membrane lipids alters the biochemical properties of the cytoplasmic membrane and enables bacteria to adapt to changes in environmental conditions. aaPGSs utilize alanine, lysine and arginine as modifying amino acids, and the primary lipid recipients have heretofore been defined as phosphatidylglycerol (PG) and cardiolipin. Here we identify a new pathway for lipid aminoacylation, conserved in many Actinobacteria, which results in formation of Ala-PG and a novel alanylated lipid, Alanyl-diacylglycerol (Ala-DAG). Ala-DAG formation in Corynebacterium glutamicum is dependent on the activity of an aaPGS homolog, whereas formation of Ala-PG requires the same enzyme acting in concert with a putative esterase encoded upstream. The presence of alanylated lipids is sufficient to enhance the bacterial fitness of C. glutamicum cultured in the presence of certain antimicrobial agents, and elucidation of this system expands the known repertoire of membrane lipids acting as substrates for amino acid modification in bacterial cells. PMID:26235234

  9. Evaluation of treadmill exercise effect on muscular lipid profiles of diabetic fatty rats by nanoflow liquid chromatography-tandem mass spectrometry.

    PubMed

    Lee, Jong Cheol; Kim, Il Yong; Son, Yeri; Byeon, Seul Kee; Yoon, Dong Hyun; Son, Jun Seok; Song, Han Sol; Song, Wook; Seong, Je Kyung; Moon, Myeong Hee

    2016-01-01

    We compare comprehensive quantitative profiling of lipids at the molecular level from skeletal muscle tissues (gastrocnemius and soleus) of Zucker diabetic fatty rats and Zucker lean control rats during treadmill exercise by nanoflow liquid chromatography-tandem mass spectrometry. Because type II diabetes is caused by decreased insulin sensitivity due to excess lipids accumulated in skeletal muscle tissue, lipidomic analysis of muscle tissues under treadmill exercise can help unveil the mechanism of lipid-associated insulin resistance. In total, 314 lipid species, including phospholipids, sphingolipids, ceramides, diacylglycerols (DAGs), and triacylglycerols (TAGs), were analyzed to examine diabetes-related lipid species and responses to treadmill exercise. Most lysophospholipid levels increased with diabetes. While DAG levels (10 from the gastrocnemius and 13 from the soleus) were >3-fold higher in diabetic rats, levels of most of these decreased after exercise in soleus but not in gastrocnemius. Levels of 5 highly abundant TAGs (52:1 and 54:3 in the gastrocnemius and 48:2, 50:2, and 52:4 in the soleus) displaying 2-fold increases in diabetic rats decreased after exercise in the soleus but not in the gastrocnemius in most cases. Thus, aerobic exercise has a stronger influence on lipid levels in the soleus than in the gastrocnemius in type 2 diabetic rats. PMID:27388225

  10. Rb-Sr and Sm-Nd Isotopic Studies of Martian Depleted Shergottes SaU 094/005

    NASA Technical Reports Server (NTRS)

    Shih, C.-Y.; Nyquist, L. E.; Reese, Y.

    2007-01-01

    Sayh al Uhaymir (SaU) 094 and SaU 005 are olivine-phyric shergottites from the Oman desert and are considered as pairs. [e.g., 1]. They are very similar to the Libyan desert shergottite Dar al Gani (DaG) 476 in petrology, chemistry and ejection age [2-6]. This group of shergottites, also recognized as depleted shergottites [e.g. 7] has been strongly shocked and contains very low abundances of light rare earth elements (REE). In addition, terrestrial contaminants are commonly present in meteorites found in desert environments. Age-dating these samples is very challenging, but lower calcite contents in the SaU meteorites suggest that they have been subjected to less severe desert weathering than their DaG counterparts [3-4]. In this report, we present Rb-Sr and Sm-Nd isotopic results for SaU 094 and SaU 005, discuss the correlation of their ages with those of other similar shergottites, and discuss their petrogenesis.

  11. Boron abundances and isotopic ratios of olivine grains on Itokawa returned by the Hayabusa spacecraft

    NASA Astrophysics Data System (ADS)

    Fujiya, Wataru; Hoppe, Peter; Ott, Ulrich

    2016-09-01

    We report the B abundances and isotopic ratios of two olivine grains from the S-type asteroid Itokawa sampled by the Hayabusa spacecraft. Olivine grains from the Dar al Gani (DaG) 989 LL6 chondrite were used as a reference. Since we analyzed polished thin sections in both cases, we expect the contribution from the solar wind B (rich in 10B) to be minimal because the solar wind was implanted only within very thin layers of the grain surface. The Itokawa and DaG 989 olivine grains have homogeneous B abundances (~400 ppb) and 11B/10B ratios compatible with the terrestrial standard and bulk chondrites. The observed homogeneous B abundances and isotopic ratios of the Itokawa olivine grains are likely the result of thermal metamorphism which occurred in the parent asteroid of Itokawa, which had a similar composition as LL chondrites. The chondritic B isotopic ratios of the Itokawa samples suggest that they contain little cosmogenic B (from cosmic-ray spallation reactions) rich in 10B. This observation is consistent with the short cosmic-ray exposure ages of Itokawa samples inferred from the small concentrations of cosmogenic 21Ne. If other Itokawa samples have little cosmogenic B as well, the enrichment in 10B found previously on the surface of another Itokawa particle (as opposed to the bulk grain study here) may be attributed to implanted solar wind B.

  12. Evaluation of treadmill exercise effect on muscular lipid profiles of diabetic fatty rats by nanoflow liquid chromatography–tandem mass spectrometry

    PubMed Central

    Lee, Jong Cheol; Kim, Il Yong; Son, Yeri; Byeon, Seul Kee; Yoon, Dong Hyun; Son, Jun Seok; Song, Han Sol; Song, Wook; Seong, Je Kyung; Moon, Myeong Hee

    2016-01-01

    We compare comprehensive quantitative profiling of lipids at the molecular level from skeletal muscle tissues (gastrocnemius and soleus) of Zucker diabetic fatty rats and Zucker lean control rats during treadmill exercise by nanoflow liquid chromatography–tandem mass spectrometry. Because type II diabetes is caused by decreased insulin sensitivity due to excess lipids accumulated in skeletal muscle tissue, lipidomic analysis of muscle tissues under treadmill exercise can help unveil the mechanism of lipid-associated insulin resistance. In total, 314 lipid species, including phospholipids, sphingolipids, ceramides, diacylglycerols (DAGs), and triacylglycerols (TAGs), were analyzed to examine diabetes-related lipid species and responses to treadmill exercise. Most lysophospholipid levels increased with diabetes. While DAG levels (10 from the gastrocnemius and 13 from the soleus) were >3-fold higher in diabetic rats, levels of most of these decreased after exercise in soleus but not in gastrocnemius. Levels of 5 highly abundant TAGs (52:1 and 54:3 in the gastrocnemius and 48:2, 50:2, and 52:4 in the soleus) displaying 2-fold increases in diabetic rats decreased after exercise in the soleus but not in the gastrocnemius in most cases. Thus, aerobic exercise has a stronger influence on lipid levels in the soleus than in the gastrocnemius in type 2 diabetic rats. PMID:27388225

  13. Differential changes in phospholipase D and phosphatidate phosphohydrolase activities in ischemia-reperfusion of rat heart.

    PubMed

    Asemu, Girma; Dent, Melissa R; Singal, Tushi; Dhalla, Naranjan S; Tappia, Paramjit S

    2005-04-01

    Phospholipase D (PLD2) produces phosphatidic acid (PA), which is converted to 1,2 diacylglycerol (DAG) by phosphatidate phosphohydrolase (PAP2). Since PA and DAG regulate Ca(2+) movements, we examined PLD2 and PAP2 in the sarcolemma (SL) and sarcoplasmic reticular (SR) membranes from hearts subjected to ischemia and reperfusion (I-R). Although SL and SR PLD2 activities were unaltered after 30 min ischemia, 5 min reperfusion resulted in a 36% increase in SL PLD2 activity, whereas 30 min reperfusion resulted in a 30% decrease in SL PLD2 activity, as compared to the control value. SR PLD2 activity was decreased (39%) after 5 min reperfusion, but returned to control levels after 30 min reperfusion. Ischemia for 60 min resulted in depressed SL and SR PLD2 activities, characterized with reduced V(max) and increased K(m) values, which were not reversed during reperfusion. Although the SL PAP2 activity was decreased (31%) during ischemia and at 30 min reperfusion (28%), the SR PAP2 activity was unchanged after 30 min ischemia, but was decreased after 5 min reperfusion (25%) and almost completely recovered after 30 min reperfusion. A 60 min period of ischemia followed by reperfusion caused an irreversible depression of SL and SR PAP2 activities. Our results indicate that I-R induced cardiac dysfunction is associated with subcellular changes in PLD2 and PAP2 activities. PMID:15752718

  14. Genetic Engineering of Dystroglycan in Animal Models of Muscular Dystrophy

    PubMed Central

    Sciandra, Francesca; Bigotti, Maria Giulia; Giardina, Bruno; Bozzi, Manuela; Brancaccio, Andrea

    2015-01-01

    In skeletal muscle, dystroglycan (DG) is the central component of the dystrophin-glycoprotein complex (DGC), a multimeric protein complex that ensures a strong mechanical link between the extracellular matrix and the cytoskeleton. Several muscular dystrophies arise from mutations hitting most of the components of the DGC. Mutations within the DG gene (DAG1) have been recently associated with two forms of muscular dystrophy, one displaying a milder and one a more severe phenotype. This review focuses specifically on the animal (murine and others) model systems that have been developed with the aim of directly engineering DAG1 in order to study the DG function in skeletal muscle as well as in other tissues. In the last years, conditional animal models overcoming the embryonic lethality of the DG knock-out in mouse have been generated and helped clarifying the crucial role of DG in skeletal muscle, while an increasing number of studies on knock-in mice are aimed at understanding the contribution of single amino acids to the stability of DG and to the possible development of muscular dystrophy. PMID:26380289

  15. The prokaryotic pathway of lipid synthesis in oat leaf plastids

    SciTech Connect

    Gillanders, B.; Mackender, R. )

    1990-05-01

    Evidence for the operation of the prokaryotic pathway of acyl lipid synthesis is an 18:3 plant was sought by incubating plastids from 6 sequential segments from oat seedling leaves as described previously except that {sup 14}C-G3P was used. Proplastids were the most biosynthetically active plastids (x2-3) and DAG and PA were the most heavily labelled acyl lipids (>75% of label). Analysis of MGDG and DGDG molecular species (MS) following pre-incubation with {sup 14}C-G3P (40 mins) and subsequently with UDP{sup 3}H-gal (30 mins), showed that only MS 16:0/18:0 and MS 16:0/18:1 were significantly labelled with {sup 14}C whereas all 5 MS were labelled with {sup 3}H-gal. However most of the {sup 3}H label was in the same MS as the {sup 14}C except when 18:2/18:2 DAG or PC was added with the {sup 3}H-gal, in which case considerably additional {sup 3}H-gal (up to 100%) appeared in the 18:2/18:2 MS of both these lipids.

  16. Transient Receptor Potential Canonical 7 (TRPC7): A Diacylglycerol-Activated Non-Selective Cation Channel

    PubMed Central

    Zhang, Xuexin

    2016-01-01

    Transient receptor potential canonical 7 (TRPC7) channel is the seventh member of the mammalian TRPC channel family. TRPC7 mRNA, protein and channel activity have been detected in many tissues and organs from mouse, rat and human. TRPC7 has high sequence homology with TRPC3 and TRPC6 and all three channels are activated by membrane receptors that couple to isoforms of phospholipase C (PLC) and mediate non-selective cation currents. TRPC7, along with TRPC3 and TRPC6 can be activated by direct exogenous application of diacylglycerol (DAG) analogs and by pharmacological maneuvers that increase endogenous DAG in cells. TRPC7 shows distinct properties of activation, such as constitutive activity, susceptibility to negative regulation by extracellular Ca2+ and by protein kinase C. TRPC7 can form heteromultimers with TRPC3 and TRPC6. Although TRPC7 remains one of the least studied TRPC channel, its role in various cell types and physiological and pathophysiological conditions is begining to emerge. PMID:24756707

  17. Lipid profiles of oil from trout (Oncorhynchus mykiss) heads, spines and viscera: trout by-products as a possible source of omega-3 lipids?

    PubMed

    Fiori, L; Solana, M; Tosi, P; Manfrini, M; Strim, C; Guella, G

    2012-09-15

    Lipid profiles of fish oil extracted from trout heads, spines and viscera using supercritical carbon dioxide and Randall extraction with hexane were measured. The amount of unsaturated fatty acids (as a percentage of total fatty acids) was within the range of 72.6-75.3% in all the substrates. A significant presence of the most important omega-3 fatty acids was detected. Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) content in oil from spines, heads and viscera resulted to be 8.7% and 7.3%, 7.9% and 6.3%, and 6.4% and 6.0%, respectively. A low (≈3%), but worth noting, presence of lipids with omega-1 polyunsaturated fatty chains was observed in all the oils. Finally, significant differences were noticed in the relative amounts of triacylglycerides (TAG), diacylglycerides (DAG) and free fatty acids (FFA). Whereas oil from heads and spines was essentially composed of TAG (≈98%), in viscera oil the molar distribution ratio became TAG:DAG:FFA=87:8:5. PMID:23107732

  18. Activation of store-operated channels by noradrenaline via protein kinase C in rabbit portal vein myocytes.

    PubMed

    Albert, A P; Large, W A

    2002-10-01

    In the present study we have investigated the role of diacylglycerol (DAG) and protein kinase C (PKC) in mediating activation of Ca(2+)-permeable store-operated channels (SOCs) by noradrenaline in rabbit portal vein smooth muscle cells. With cell-attached recording, bath application of noradrenaline, 1-oleoyl-acetyl-sn-glycerol (OAG) and phorbol 12,13-dibutyrate (PDBu) evoked single channel currents. The biophysical properties of these channel currents were similar to those of the channel currents activated by depletion of internal Ca(2+) stores with cyclopiazonic acid (CPA). The activation of SOCs in cell-attached recording by noradrenaline, OAG, PDBu, CPA and the acetoxymethyl ester form of BAPTA (BAPTA-AM) was markedly inhibited by the PKC inhibitors chelerythrine and RO-31-8220. In isolated outside-out patches CPA did not evoke SOCs but noradrenaline stimulated SOC activity, which was reduced by about 90 % by PKC inhibitors. The addition of the serine/threonine phosphatase inhibitors calyculin A and microcystin also stimulated SOCs in isolated outside-out patches. It is concluded that in rabbit portal vein myocytes, noradrenaline activates SOCs via DAG and PKC, possibly by a store-independent mechanism. In addition in this cell type it appears that PKC and phosphorylation may play an important role in stimulating SOC activity in response to depletion of internal Ca(2+) stores by CPA and BAPTA-AM. PMID:12356885

  19. Trans-activation of TRPV1 by D1R in mouse dorsal root ganglion neurons.

    PubMed

    Lee, Dong Woo; Cho, Pyung Sun; Lee, Han Kyu; Lee, Sang Hoon; Jung, Sung Jun; Oh, Seog Bae

    2015-10-01

    TRPV1, a ligand-gated ion channel expressed in nociceptive sensory neurons is modulated by a variety of intracellular signaling pathways. Dopamine is a neurotransmitter that plays important roles in motor control, cognition, and pain modulation in the CNS, and acts via a variety of dopamine receptors (D1R-D5R), a class of GPCRs. Although nociceptive sensory neurons express D1-like receptors, very little is known about the effect of dopamine on TRPV1 in the peripheral nervous system. Therefore, in this study, we examined the effects of D1R activation on TRPV1 in mouse DRG neurons using Ca(2+) imaging and immunohistochemical analysis. The D1R agonist SKF-38393 induced reproducible Ca(2+) responses via Ca(2+) influx through TRPV1 rather than Ca(2+) mobilization from intracellular Ca(2+) stores. Immunohistochemical analysis revealed co-expression of D1R and TRPV1 in mouse DRG neurons. The PLC-specific inhibitor blocked the SKF-38393-induced Ca(2+) response, whereas the PKC, DAG lipase, AC, and PKA inhibitors had no effect on the SKF-38393-induced Ca(2+) response. Taken together, our results suggest that the SKF-38393-induced Ca(2+) response results from the direct activation of TRPV1 by a PLC/DAG-mediated membrane-delimited pathway. These results provide evidence that the trans-activation of TRPV1 following D1R activation may contribute to the modulation of pain signaling in nociceptive sensory neurons. PMID:26319554

  20. Potential role of salicylic acid in modulating diacylglycerol homeostasis in response to freezing temperatures in Arabidopsis

    PubMed Central

    Tan, Wei-Juan; Xiao, Shi; Chen, Qin-Fang

    2015-01-01

    In our recent article in Molecular Plant, we reported that 3 lipase-like defense regulators SENESCENCE-ASSOCIATED GENE101 (SAG101), ENHANCED DISEASE SUSCEPTIBILITY1 (EDS1) and PHYTOALEXIN DEFICIENT4 (PAD4) are involved in the regulation of freezing tolerance in Arabidopsis. The transcripts of SAG101, EDS1 and PAD4 were inducible by cold stress and their knockout or knockdown mutants exhibited enhanced chilling and freezing tolerance in comparison to the wild type. The freezing tolerance phenotype showed in the sag101, eds1 and pad4 mutants was correlated with the transcriptional upregulation of C-REPEAT/DRE BINDING FACTORs (CBFs) and their regulons as well as increased levels of proline. Upon cold exposure, the sag101, eds1 and pad4 mutants showed ameliorated cell death and accumulation of hydrogen peroxide, which were highly induced by freezing stress in the wild-type leaves. Moreover, the contents of salicylic acid (SA) and diacylglycerol (DAG) were significantly decreased in the sag101, eds1 and pad4 mutants compared to the wild type. Taken together, our results suggest that the SAG101, EDS1 and PAD4 are negative regulators in the freezing response and function, at least in part, by modulating the homeostasis of SA and DAG in Arabidopsis. PMID:26340231

  1. Other factors to consider in the formation of chloropropandiol fatty esters in oil processes.

    PubMed

    Ramli, Muhamad Roddy; Siew, Wai Lin; Ibrahim, Nuzul Amri; Kuntom, Ainie; Abd Razak, Raznim Arni

    2015-01-01

    This paper examines the processing steps of extracting palm oil from fresh fruit bunches in a way that may impact on the formation of chloropropandiol fatty esters (3-MCPD esters), particularly during refining. Diacylglycerols (DAGs) do not appear to be a critical factor when crude palm oils are extracted from various qualities of fruit bunches. Highly hydrolysed oils, in spite of the high free fatty acid (FFA) contents, did not show exceptionally high DAGs, and the oils did not display a higher formation of 3-MCPD esters upon heat treatment. However, acidity measured in terms of pH appears to have a strong impact on 3-MCPD ester formation in the crude oil when heated at high temperatures. The differences in the extraction process of crude palm oil from current commercial processes and that from a modified experimental process showed clearly the effect of acidity of the oil on the formation of 3-MCPD esters. This paper concludes that the washing or dilution step in palm oil mills removes the acidity of the vegetative materials and that a well-optimised dilution/washing step in the extraction process will play an important role in reducing formation of 3-MCPD esters in crude palm oil upon further heat processing. PMID:25798697

  2. Changes in metabolic pathways of Desulfovibrio alaskensis G20 cells induced by molybdate excess.

    PubMed

    Nair, Rashmi R; Silveira, Célia M; Diniz, Mário S; Almeida, Maria G; Moura, Jose J G; Rivas, Maria G

    2015-03-01

    The activity of sulfate-reducing bacteria (SRB) intensifies the problems associated to corrosion of metals and the solution entails significant economic costs. Although molybdate can be used to control the negative effects of these organisms, the mechanisms triggered in the cells exposed to Mo-excess are poorly understood. In this work, the effects of molybdate ions on the growth and morphology of the SRB Desulfovibrio alaskensis G20 (DaG20) were investigated. In addition, the cellular localization, ion uptake and regulation of protein expression were studied. We found that molybdate concentrations ranging between 50 and 150 µM produce a twofold increase in the doubling time with this effect being more significant at 200 µM molybdate (five times increase in the doubling time). It was also observed that 500 µM molybdate completely inhibits the cellular growth. On the context of protein regulation, we found that several enzymes involved in energy metabolism, cellular division and metal uptake processes were particularly influenced under the conditions tested. An overall description of some of the mechanisms involved in the DaG20 adaptation to molybdate-stress conditions is discussed. PMID:25488518

  3. Formation of Glycidyl Fatty Acid Esters Both in Real Edible Oils during Laboratory-Scale Refining and in Chemical Model during High Temperature Exposure.

    PubMed

    Cheng, Weiwei; Liu, Guoqin; Liu, Xinqi

    2016-07-27

    In the present study, the formation mechanisms of glycidyl fatty acid esters (GEs) were investigated both in real edible oils (soybean oil, camellia oil, and palm oil) during laboratory-scale preparation and refining and in chemical model (1,2-dipalmitin (DPG) and 1-monopalmitin (MPG)) during high temperature exposure (160-260 °C under nitrogen). The formation process of GEs in the chemical model was monitored using attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy. The results showed that the roasting and pressing process could produce certain amounts of GEs that were much lower than that produced in the deodorization process. GE contents in edible oils increased continuously and significantly with increasing deodorization time below 200 °C. However, when the temperature exceeded 200 °C, GE contents sharply increased in 1-2 h followed by a gradual decrease, which could verify a simultaneous formation and degradation of GEs at high temperature. In addition, it was also found that the presence of acylglycerol (DAGs and MAGs) could significantly increase the formation yield of GEs both in real edible oils and in chemical model. Compared with DAGs, moreover, MAGs displayed a higher formation capacity but substantially lower contribution to GE formation due to their low contents in edible oils. In situ ATR-FTIR spectroscopic evidence showed that cyclic acyloxonium ion intermediate was formed during GE formation derived from DPG and MPG in chemical model heated at 200 °C. PMID:27319409

  4. Muscle ERRγ mitigates Duchenne muscular dystrophy via metabolic and angiogenic reprogramming.

    PubMed

    Matsakas, Antonios; Yadav, Vikas; Lorca, Sabina; Narkar, Vihang

    2013-10-01

    Treatment of Duchenne muscular dystrophy (DMD) by replacing mutant dystrophin or restoring dystrophin-associated glycoprotein complex (DAG) has been clinically challenging. Instead, identifying and targeting muscle pathways deregulated in DMD will provide new therapeutic avenues. We report that the expression of nuclear receptor estrogen-related receptor-γ (ERRγ), and its metabolic and angiogenic targets are down-regulated (50-85%) in skeletal muscles of mdx mice (DMD model) vs. wild-type mice. Corelatively, oxidative myofibers, muscle vasculature, and exercise tolerance (33%) are decreased in mdx vs. wild-type mice. Overexpressing ERRγ selectively in the dystrophic muscles of the mdx mice restored metabolic and angiogenic gene expression compared with control mdx mice. Further, ERRγ enhanced muscle oxidative myofibers, vasculature, and blood flow (by 33-66%) and improved exercise tolerance (by 75%) in the dystrophic mice. Restoring muscle ERRγ pathway ameliorated muscle damage and also prevented DMD hallmarks of postexercise muscle damage, hypoxia, and fatigue in mdx mice. Notably, ERRγ did not restore sarcolemmal DAG complex, which is thus dispensable for antidystrophic effects of ERRγ. In summary, ERRγ-dependent metabolic and angiogenic gene program is defective in DMD, and we demonstrate that its restoration is a potential strategy for treating muscular dystrophy. PMID:23781095

  5. Recombination-aware alignment of diploid individuals

    PubMed Central

    2014-01-01

    Background Traditionally biological similarity search has been studied under the abstraction of a single string to represent each genome. The more realistic representation of diploid genomes, with two strings defining the genome, has so far been largely omitted in this context. With the development of sequencing techniques and better phasing routines through haplotype assembly algorithms, we are not far from the situation when individual diploid genomes could be represented in their full complexity with a pair-wise alignment defining the genome. Results We propose a generalization of global alignment that is designed to measure similarity between phased predictions of individual diploid genomes. This generalization takes into account that individual diploid genomes evolve through a mutation and recombination process, and that predictions may be erroneous in both dimensions. Even though our model is generic, we focus on the case where one wants to measure only the similarity of genome content allowing free recombination. This results into efficient algorithms for direct application in (i) evaluation of variation calling predictions and (ii) progressive multiple alignments based on labeled directed acyclic graphs (DAGs) to represent profiles. The latter may be of more general interest, in connection to covering alignment of DAGs. Extensions of our model and algorithms can be foreseen to have applications in evaluating phasing algorithms, as well as more fundamental role in phasing child genome based on parent genomes. PMID:25572943

  6. Roles of phosphoinositide-specific phospholipase Cγ1 in brain development.

    PubMed

    Kang, Du-Seock; Yang, Yong Ryoul; Lee, Cheol; Kim, SaetByeol; Ryu, Sung Ho; Suh, Pann-Ghill

    2016-01-01

    Over the past decade, converging evidence suggests that PLCγ1 signaling has key roles in controlling neural development steps. PLCγ1 functions as a signal transducer that converts an extracellular stimulus into intracellular signals by generating second messengers such as DAG and IP3. DAG functions as an activator of either PKC or transient receptor potential cation channels (TRPCs), while IP3 induces the calcium release from intracellular calcium stores. These second messengers regulate the morphological change of neuron, such as neurite outgrowth, migration, axon pathfinding, and synapse formation. These morphological changes depend on finely tuned calcium signaling following receptor tyrosine kinase-mediated PLCγ1 signaling. Thus, deregulation of PLCγ1 signaling causes various abnormalities of neuronal development and it may be associated with diverse neurological disorders. Herein, we discuss the current understanding of the PLCγ1 signaling pathway in neural development and provide recent advances of how PLCγ1 signaling is involved in the formation of neuronal processes for functionally faithful brain development. PMID:26588873

  7. Model Averaging for Improving Inference from Causal Diagrams

    PubMed Central

    Hamra, Ghassan B.; Kaufman, Jay S.; Vahratian, Anjel

    2015-01-01

    Model selection is an integral, yet contentious, component of epidemiologic research. Unfortunately, there remains no consensus on how to identify a single, best model among multiple candidate models. Researchers may be prone to selecting the model that best supports their a priori, preferred result; a phenomenon referred to as “wish bias”. Directed acyclic graphs (DAGs), based on background causal and substantive knowledge, are a useful tool for specifying a subset of adjustment variables to obtain a causal effect estimate. In many cases, however, a DAG will support multiple, sufficient or minimally-sufficient adjustment sets. Even though all of these may theoretically produce unbiased effect estimates they may, in practice, yield somewhat distinct values, and the need to select between these models once again makes the research enterprise vulnerable to wish bias. In this work, we suggest combining adjustment sets with model averaging techniques to obtain causal estimates based on multiple, theoretically-unbiased models. We use three techniques for averaging the results among multiple candidate models: information criteria weighting, inverse variance weighting, and bootstrapping. We illustrate these approaches with an example from the Pregnancy, Infection, and Nutrition (PIN) study. We show that each averaging technique returns similar, model averaged causal estimates. An a priori strategy of model averaging provides a means of integrating uncertainty in selection among candidate, causal models, while also avoiding the temptation to report the most attractive estimate from a suite of equally valid alternatives. PMID:26270672

  8. Dynamic Uncertain Causality Graph for Knowledge Representation and Probabilistic Reasoning: Directed Cyclic Graph and Joint Probability Distribution.

    PubMed

    Zhang, Qin

    2015-07-01

    Probabilistic graphical models (PGMs) such as Bayesian network (BN) have been widely applied in uncertain causality representation and probabilistic reasoning. Dynamic uncertain causality graph (DUCG) is a newly presented model of PGMs, which can be applied to fault diagnosis of large and complex industrial systems, disease diagnosis, and so on. The basic methodology of DUCG has been previously presented, in which only the directed acyclic graph (DAG) was addressed. However, the mathematical meaning of DUCG was not discussed. In this paper, the DUCG with directed cyclic graphs (DCGs) is addressed. In contrast, BN does not allow DCGs, as otherwise the conditional independence will not be satisfied. The inference algorithm for the DUCG with DCGs is presented, which not only extends the capabilities of DUCG from DAGs to DCGs but also enables users to decompose a large and complex DUCG into a set of small, simple sub-DUCGs, so that a large and complex knowledge base can be easily constructed, understood, and maintained. The basic mathematical definition of a complete DUCG with or without DCGs is proved to be a joint probability distribution (JPD) over a set of random variables. The incomplete DUCG as a part of a complete DUCG may represent a part of JPD. Examples are provided to illustrate the methodology. PMID:25781960

  9. The LHCb DIRAC-based production and data management operations systems

    NASA Astrophysics Data System (ADS)

    Stagni, F.; Charpentier, P.; LHCb Collaboration

    2012-06-01

    The LHCb computing model was designed in order to support the LHCb physics program, taking into account LHCb specificities (event sizes, processing times etc...). Within this model several key activities are defined, the most important of which are real data processing (reconstruction, stripping and streaming, group and user analysis), Monte-Carlo simulation and data replication. In this contribution we detail how these activities are managed by the LHCbDIRAC Data Transformation System. The LHCbDIRAC Data Transformation System leverages the workload and data management capabilities provided by DIRAC, a generic community grid solution, to support data-driven workflows (or DAGs). The ability to combine workload and data tasks within a single DAG allows to create highly sophisticated workflows with the individual steps linked by the availability of data. This approach also provides the advantage of a single point at which all activities can be monitored and controlled. While several interfaces are currently supported (including python API and CLI), we will present the ability to create LHCb workflows through a secure web interface, control their state in addition to creating and submitting jobs. To highlight the versatility of the system we present in more detail experience with real data of the 2010 and 2011 LHC run.

  10. Natural isoflavones regulate the quadruplex-duplex competition in human telomeric DNA.

    PubMed

    Zhang, Jin-li; Fu, Yan; Zheng, Lin; Li, Wei; Li, Hao; Sun, Qian; Xiao, Ying; Geng, Feng

    2009-05-01

    Effects of natural isoflavones on the structural competition of human telomeric G-quadruplex d[AG(3)(T(2)AG(3))(3)] and its related Watson-Crick duplex d[AG(3)(T(2)AG(3))(3)-(C(3)TA(2))(3)C(3)T] are investigated by using circular dichroism (CD), ESI-MS, fluorescence quenching measurement, CD stopped-flow kinetic experiment, UV spectroscopy and molecular modeling methods. It is intriguing to find out that isoflavones can stabilize the G-quadruplex structure but destabilize its corresponding Watson-Crick duplex and this discriminated interaction is intensified by molecular crowding environments. Kinetic experiments indicate that the dissociation rate of quadruplex (k(obs290 nm)) is decreased by 40.3% at the daidzin/DNA molar ratio of 1.0 in K(+), whereas in Na(+) the observed rate constant is reduced by about 12.0%. Furthermore, glycosidic daidzin significantly induces a structural transition of the polymorphic G-quadruplex into the antiparallel conformation in K(+). This is the first report on the recognition of isoflavones with conformational polymorphism of G-quadruplex, which suggests that natural isoflavone constituents potentially exhibit distinct regulation on the structural competition of quadruplex versus duplex in human telomeric DNA. PMID:19261597

  11. Acid sphingomyelinase activity is regulated by membrane lipids and facilitates cholesterol transfer by NPC2[S

    PubMed Central

    Oninla, Vincent O.; Breiden, Bernadette; Babalola, Jonathan O.; Sandhoff, Konrad

    2014-01-01

    During endocytosis, membrane components move to intraluminal vesicles of the endolysosomal compartment for digestion. At the late endosomes, cholesterol is sorted out mainly by two sterol-binding proteins, Niemann-Pick protein type C (NPC)1 and NPC2. To study the NPC2-mediated intervesicular cholesterol transfer, we developed a liposomal assay system. (Abdul-Hammed, M., B. Breiden, M. A. Adebayo, J. O. Babalola, G. Schwarzmann, and K. Sandhoff. 2010. Role of endosomal membrane lipids and NPC2 in cholesterol transfer and membrane fusion. J. Lipid Res. 51: 1747–1760.) Anionic lipids stimulate cholesterol transfer between liposomes while SM inhibits it, even in the presence of anionic bis(monoacylglycero)phosphate (BMP). Preincubation of vesicles containing SM with acid sphingomyelinase (ASM) (SM phosphodiesterase, EC 3.1.4.12) results in hydrolysis of SM to ceramide (Cer), which enhances cholesterol transfer. Besides SM, ASM also cleaves liposomal phosphatidylcholine. Anionic phospholipids derived from the plasma membrane (phosphatidylglycerol and phosphatidic acid) stimulate SM and phosphatidylcholine hydrolysis by ASM more effectively than BMP, which is generated during endocytosis. ASM-mediated hydrolysis of liposomal SM was also stimulated by incorporation of diacylglycerol (DAG), Cer, and free fatty acids into the liposomal membranes. Conversely, phosphatidylcholine hydrolysis was inhibited by incorporation of cholesterol, Cer, DAG, monoacylglycerol, and fatty acids. Our data suggest that SM degradation by ASM is required for physiological secretion of cholesterol from the late endosomal compartment, and is a key regulator of endolysosomal lipid digestion. PMID:25339683

  12. Biosynthesis of lipid a precursors: cloning, expression and partial purification of E. coli UDP-GlcNAc acyl transferase; identification of UDP-3-O-acyl-GlcNAc as its enzymatic product

    SciTech Connect

    Anderson, M.S.; Crowell, D.N.; Schlafmann, S.E.; Raetz, C.R.H.

    1986-05-01

    The initial steps in the biosynthesis of E. coli lipid A disaccharides have been shown to proceed by two acylations of UDP-GlcNAc to form UDP-2,3-diacyl-GlcN (UDP-DAG). In vitro synthesis of UDP-DAG appears to involve a previously undescribed, monoacylated form of UDP-GlcNAc. The authors now show (1) definitive evidence that this metabolite has the structure UDP-3-O-acyl-GlcNAc, and (2) that a gene encoding or controlling this acylating enzyme is located on a 2.4 Kb fragment mapping between dnaE and cds on the E. coli chromosome. Insertion of this fragment into an expression vector directs overproduction of an enzyme catalyzing monoacylation of UDP-GlcNAc by acyl-acyl carrier protein. The activity was partially purified from an overproducing strain by chromatography of the soluble fraction of a cell extract on DEAE cellulose in the presence of 1% octyl glucoside. Peak fractions were 2200x purified relative to wild type with an 80% yield. The product generated by this enzyme was isolated from a preparative scale incubation and structurally identified by /sup 1/H-NMR.

  13. Summary of the activities of the subgroup on data acquisition and processing

    SciTech Connect

    Connolly, P.L.; Doughty, D.C.; Elias, J.E.

    1981-01-01

    A data acquisition and handling subgroup consisting of approximately 20 members met during the 1981 ISABELLE summer study. Discussions were led by members of the BNL ISABELLE Data Acquisition Group (DAG) with lively participation from outside users. Particularly large contributions were made by representatives of BNL experiments 734, 735, and the MPS, as well as the Fermilab Colliding Detector Facility and the SLAC LASS Facility. In contrast to the 1978 study, the subgroup did not divide its activities into investigations of various individual detectors, but instead attempted to review the current state-of-the-art in the data acquisition, trigger processing, and data handling fields. A series of meetings first reviewed individual pieces of the problem, including status of the Fastbus Project, the Nevis trigger processor, the SLAC 168/E and 3081/E emulators, and efforts within DAG. Additional meetings dealt with the question involving specifying and building complete data acquisition systems. For any given problem, a series of possible solutions was proposed by the members of the subgroup. In general, any given solution had both advantages and disadvantages, and there was never any consensus on which approach was best. However, there was agreement that certain problems could only be handled by systems of a given power or greater. what will be given here is a review of various solutions with associated powers, costs, advantages, and disadvantages.

  14. Plasma diacylglycerol composition is a biomarker of metabolic syndrome onset in rhesus monkeys.

    PubMed

    Polewski, Michael A; Burhans, Maggie S; Zhao, Minghui; Colman, Ricki J; Shanmuganayagam, Dhanansayan; Lindstrom, Mary J; Ntambi, James M; Anderson, Rozalyn M

    2015-08-01

    Metabolic syndrome is linked with obesity and is often first identified clinically by elevated BMI and elevated levels of fasting blood glucose that are generally secondary to insulin resistance. Using the highly translatable rhesus monkey (Macaca mulatta) model, we asked if metabolic syndrome risk could be identified earlier. The study involved 16 overweight but healthy, euglycemic monkeys, one-half of which spontaneously developed metabolic syndrome over the course of 2 years while the other half remained healthy. We conducted a series of biometric and plasma measures focusing on adiposity, lipid metabolism, and adipose tissue-derived hormones, which led to a diagnosis of metabolic syndrome in the insulin-resistant animals. Plasma fatty acid composition was determined by gas chromatography for cholesteryl ester, FFA, diacylglycerol (DAG), phospholipid, and triacylglycerol lipid classes; plasma lipoprotein profiles were generated by NMR; and circulating levels of adipose-derived signaling peptides were determined by ELISA. We identified biomarker models including a DAG model, two lipoprotein models, and a multiterm model that includes the adipose-derived peptide adiponectin. Correlations among circulating lipids and lipoproteins revealed shifts in lipid metabolism during disease development. We propose that lipid profiling may be valuable for early metabolic syndrome detection in a clinical setting. PMID:26063458

  15. RasGRP3 regulates the migration of glioma cells via interaction with Arp3

    PubMed Central

    Lee, Hae Kyung; Finniss, Susan; Cazacu, Simona; Xiang, Cunli; Poisson, Laila M.; Blumberg, Peter M.; Brodie, Chaya

    2015-01-01

    Glioblastoma (GBM), the most aggressive primary brain tumors, are highly infiltrative. Although GBM express high Ras activity and Ras proteins have been implicated in gliomagenesis, Ras-activating mutations are not frequent in these tumors. RasGRP3, an important signaling protein responsive to diacylglycerol (DAG), increases Ras activation. Here, we examined the expression and functions of RasGRP3 in GBM and glioma cells. RasGRP3 expression was upregulated in GBM specimens and glioma stem cells compared with normal brains and neural stem cells, respectively. RasGRP3 activated Ras and Rap1 in glioma cells and increased cell migration and invasion partially via Ras activation. Using pull-down assay and mass spectroscopy we identified the actin-related protein, Arp3, as a novel interacting protein of RasGRP3. The interaction of RasGRP3 and Arp3 was validated by immunofluorescence staining and co-immunoprecipitation, and PMA, which activates RasGRP3 and induces its translocation to the peri-nuclear region, increased the association of Arp3 and RasGRP3. Arp3 was upregulated in GBM, regulated cell spreading and migration and its silencing partially decreased these effects of RasGRP3 in glioma cells. In summary, RasGRP3 acts as an important integrating signaling protein of the DAG and Ras signaling pathways and actin polymerization and represents an important therapeutic target in GBM. PMID:25682201

  16. Fenofibrate insulates diacylglycerol in lipid droplet/ER and preserves insulin signaling transduction in the liver of high fat fed mice.

    PubMed

    Chan, Stanley M H; Zeng, Xiao-Yi; Sun, Ruo-Qiong; Jo, Eunjung; Zhou, Xiu; Wang, Hao; Li, Songpei; Xu, Aimin; Watt, Matthew J; Ye, Ji-Ming

    2015-07-01

    Hepatic steatosis is often associated with insulin resistance as a hallmark of the metabolic syndrome in the liver. The present study investigated the effects of PPARα activation induced by fenofibrate (FB) on the relationship of insulin resistance and hepatic steatosis in mice fed a high-fat (HF) diet, which increases lipid influx into the liver. Mice were fed HF diet to induce insulin resistance and hepatic steatosis with or without FB. FB activated PPARα and ameliorated HF diet-induced glucose intolerance and hepatic insulin resistance without altering either hepatic steatosis or inflammation signaling (JNK or IKK). Interestingly, FB treatment simultaneously increased fatty acid (FA) synthesis (50%) and oxidation (66%, both p<0.01) into intermediate lipid metabolites, suggesting a FA oxidation-synthesis cycling in operation. Associated with these effects, diacylglycerols (DAGs) were sequestered within the lipid droplet/ER compartment, thus reducing their deposition in the cellular membrane, which is known to impair insulin signal transduction. These findings suggest that the reduction in membrane DAGs (rather than total hepatic steatosis) may be critical for the protection by fenofibrate-induced PPARα activation against hepatic insulin resistance induced by dietary fat. PMID:25906681

  17. Various Acylglycerols from Common Oils Exert Different Antitumor Activities on Colorectal Cancer Cells.

    PubMed

    Ramos-Bueno, Rebeca P; González-Fernández, María J; Guil-Guerrero, José L

    2016-01-01

    Colorectal cancer is one of the leading causes of death in Western countries; therefore, the implementation of healthy dietary habits in order to prevent its occurrence is a desirable action. We show here that both free fatty acids (FFAs) and some acylglycerols induce antitumoral actions in the colorectal cancer cell line HT-29. We tested several C18 polyunsaturated fatty acid-enriched oils (e.g., sunflower and Echium) as well as other oils, such as arachidonic acid-enriched (Arasco®) and docosahexaenoic acid-enriched (Marinol® and cod liver oil), in addition to coconut and olive oils. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test indicated inhibitory effects on HT-29 cells viability for FFAs, and monoacylglycerol and diacylglycerol (DAG) species, while the lactate dehydrogenase test proved that FFAs were the more effective species to induce membrane injury. Conversely, all species did not exhibit actions on CCD-18 normal human colon cells viability. Furthermore, transmission electron microscopy showed the presence of necrosis and apoptosis, while the monoacylglycerol lipase (MAGL) inhibition test demonstrated high activity for 2-monoacylglycerols derived from Arasco and sunflower oils. However, different monoacylglycerols and DAGs have also the potential for MAGL inhibition. Therefore, checking for activity on colon cancer cells of specifically designed acylglycerol-derivative species would be a suitable way to design functional foods destined to avoid colorectal cancer initiation. PMID:27007804

  18. PKC alpha mediates maternal touch regulation of growth-related gene expression in infant rats.

    PubMed

    Schanberg, Saul M; Ingledue, Vickie F; Lee, Joanna Y; Hannun, Yusuf A; Bartolome, Jorge V

    2003-06-01

    During short-term periods of separation of rat pups from their mothers, the loss of certain sensory signals suppresses the increase in ornithine decarboxylase (ODC) gene expression induced by the growth-promoting hormones prolactin (PRL) and growth hormone (GH). Here, we identify a molecular mechanism through which maternal separation (MS) curtails ODC expression. Our results demonstrate that the absence of specific tactile stimuli provided by the mother limits PRL-evoked stimulation of ODC biosynthesis by interfering with sn-1,2-diacylglycerol's (DAG) ability to activate protein kinase Calpha (PKCalpha) and consequently c-myc mRNA and max mRNA expression. The proteins encoded by these proto-oncogenes function as direct transactivators of the ODC gene. As ODC activity is obligatory for normal cell replication and differentiation, PKCalpha activation by DAG represents an important control point at which 'nurturing touch' regulates growth and development of the neonate. Such a mechanism can explain the maladaptive consequences of disrupting mother-infant tactile interactions as occurs in isolated premature babies. Also, it could provide a basis for developing therapeutic interventions to maximize growth potential in children failing-to-thrive despite normal maternal care. PMID:12700701

  19. A soluble diacylglycerol acyltransferase is involved in triacylglycerol biosynthesis in the oleaginous yeast Rhodotorula glutinis.

    PubMed

    Rani, Sapa Hima; Saha, Saikat; Rajasekharan, Ram

    2013-01-01

    The biosynthesis of triacylglycerol (TAG) occurs in the microsomal membranes of eukaryotes. Here, we report the identification and functional characterization of diacylglycerol acyltransferase (DGAT), a member of the 10 S cytosolic TAG biosynthetic complex (TBC) in Rhodotorula glutinis. Both a full-length and an N-terminally truncated cDNA clone of a single gene were isolated from R. glutinis. The DGAT activity of the protein encoded by RgDGAT was confirmed in vivo by the heterologous expression of cDNA in a Saccharomyces cerevisiae quadruple mutant (H1246) that is defective in TAG synthesis. RgDGAT overexpression in yeast was found to be capable of acylating diacylglycerol (DAG) in an acyl-CoA-dependent manner. Quadruple mutant yeast cells exhibit growth defects in the presence of oleic acid, but wild-type yeast cells do not. In an in vivo fatty acid supplementation experiment, RgDGAT expression rescued quadruple mutant growth in an oleate-containing medium. We describe a soluble acyl-CoA-dependent DAG acyltransferase from R. glutinis that belongs to the DGAT3 class of enzymes. The study highlights the importance of an alternative TAG biosynthetic pathway in oleaginous yeasts. PMID:23103975

  20. Muscular dystrophy associated with alpha-dystroglycan deficiency in Sphynx and Devon Rex cats.

    PubMed

    Martin, Paul T; Shelton, G Diane; Dickinson, Peter J; Sturges, Beverly K; Xu, Rui; LeCouteur, Richard A; Guo, Ling T; Grahn, Robert A; Lo, Harriet P; North, Kathryn N; Malik, Richard; Engvall, Eva; Lyons, Leslie A

    2008-12-01

    Recent studies have identified a number of forms of muscular dystrophy, termed dystroglycanopathies, which are associated with loss of natively glycosylated alpha-dystroglycan. Here we identify a new animal model for this class of disorders in Sphynx and Devon Rex cats. Affected cats displayed a slowly progressive myopathy with clinical and histologic hallmarks of muscular dystrophy including skeletal muscle weakness with no involvement of peripheral nerves or CNS. Skeletal muscles had myopathic features and reduced expression of alpha-dystroglycan, while beta-dystroglycan, sarcoglycans, and dystrophin were expressed at normal levels. In the Sphynx cat, analysis of laminin and lectin binding capacity demonstrated no loss in overall glycosylation or ligand binding for the alpha-dystroglycan protein, only a loss of protein expression. A reduction in laminin-alpha2 expression in the basal lamina surrounding skeletal myofibers was also observed. Sequence analysis of translated regions of the feline dystroglycan gene (DAG1) in affected cats did not identify a causative mutation, and levels of DAG1 mRNA determined by real-time QRT-PCR did not differ significantly from normal controls. Reduction in the levels of glycosylated alpha-dystroglycan by immunoblot was also identified in an affected Devon Rex cat. These data suggest that muscular dystrophy in Sphynx and Devon Rex cats results from a deficiency in alpha-dystroglycan protein expression, and as such may represent a new type of dystroglycanopathy where expression, but not glycosylation, is affected. PMID:18990577

  1. Inositolphosphoglycans and diacyglycerol are possible mediators in the glycogenic effect of GLP-1(7-36)amide in BC3H-1 myocytes.

    PubMed

    Galera, C; Clemente, F; Alcantara, A; Trapote, M A; Perea, A; Lopez-Delgado, M I; Villanueva-Penacarrillo, M L; Valverde, I

    1996-03-01

    A potent glycogenic effect of GLP-1(7-36)amide has been found in rat hepatocytes and skeletal muscle, and specific receptors for this peptide, which do not seem to be associated with the adenylate cyclase-cAMP system, have been detected in these tissue membranes. On the other hand, inositolphosphoglycan molecules (IPGs) have been implicated as second messengers of the action of insulin. In this work, we have found, in differentiated BC3H-1 myocytes, specific binding of [125I]GLP-1(7-36)amide, and a stimulatory effect of the peptide on glycogen synthesis, confirming the findings in rat skeletal muscle. Also, GLP-1(7-36)amide modulates the cell content of radiolabelled glycosylphosphatidylinositols (GPIs) and increases the production of diacylglycerol (DAG), in the same manner as insulin acts, indicating hydrolysis of GPIs and an immediate and short-lived generation of IPGs. Thus, IPGs and DAG could be mediators in the glycogenic action of GLP-1(7-36)amide in skeletal muscle. PMID:8907253

  2. Myxococcus CsgA, Drosophila Sniffer, and human HSD10 are cardiolipin phospholipases

    PubMed Central

    Boynton, Tye O'Hara; Shimkets, Lawrence Joseph

    2015-01-01

    Myxococcus xanthus development requires CsgA, a member of the short-chain alcohol dehydrogenase (SCAD) family of proteins. We show that CsgA and SocA, a protein that can replace CsgA function in vivo, oxidize the 2′-OH glycerol moiety on cardiolipin and phosphatidylglycerol to produce diacylglycerol (DAG), dihydroxyacetone, and orthophosphate. A lipid extract enriched in DAGs from wild-type cells initiates development and lipid body production in a csgA mutant to bypass the mutational block. This novel phospholipase C-like reaction is widespread. SCADs that prevent neurodegenerative disorders, such as Drosophila Sniffer and human HSD10, oxidize cardiolipin with similar kinetic parameters. HSD10 exhibits a strong preference for cardiolipin with oxidized fatty acids. This activity is inhibited in the presence of the amyloid β peptide. Three HSD10 variants associated with neurodegenerative disorders are inactive with cardiolipin. We suggest that HSD10 protects humans from reactive oxygen species by removing damaged cardiolipin before it induces apoptosis. PMID:26338420

  3. Recessive mutations in DGKE cause atypical hemolytic-uremic syndrome

    PubMed Central

    Lemaire, Mathieu; Frémeaux-Bacchi, Véronique; Schaefer, Franz; Choi, Murim; Tang, Wai Ho; Le Quintrec, Moglie; Fakhouri, Fadi; Taque, Sophie; Nobili, François; Martinez, Frank; Ji, Weizhen; Overton, John D.; Mane, Shrikant M.; Nürnberg, Gudrun; Altmüller, Janine; Thiele, Holger; Morin, Denis; Deschenes, Georges; Baudouin, Véronique; Llanas, Brigitte; Collard, Laure; Majid, Mohammed A.; Simkova, Eva; Nürnberg, Peter; Rioux-Leclerc, Nathalie; Moeckel, Gilbert W.; Gubler, Marie Claire; Hwa, John; Loirat, Chantal; Lifton, Richard P.

    2013-01-01

    Pathologic thrombosis is a major cause of mortality. Hemolytic-uremic syndrome (HUS) features episodes of small vessel thrombosis resulting in microangiopathic hemolytic anemia, thrombocytopenia and renal failure1. Atypical HUS (aHUS) can result from genetic or autoimmune factors2 that lead to pathologic complement cascade activation3. By exome sequencing we identify recessive mutations in DGKE (diacylglycerol kinase epsilon) that co-segregate with aHUS in 9 unrelated kindreds, defining a distinctive Mendelian disease. Affected patients present with aHUS before age 1, have persistent hypertension, hematuria and proteinuria (sometimes nephrotic range), and develop chronic kidney disease with age. DGKE is found in endothelium, platelets, and podocytes. Arachidonic acid-containing diacylglycerols (DAG) activate protein kinase C, which promotes thrombosis. DGKE normally inactivates DAG signaling. We infer that loss of DGKE function results in a pro-thrombotic state. These findings identify a new mechanism of pathologic thrombosis and kidney failure and have immediate implications for treatment of aHUS patients. PMID:23542698

  4. Differential Distortion of Purine Substrates by Human and Plasmodium falciparum Hypoxanthine-Guanine Phosphoribosyltransferase to Catalyse the Formation of Mononucleotides.

    PubMed

    Karnawat, Vishakha; Gogia, Spriha; Balaram, Hemalatha; Puranik, Mrinalini

    2015-07-20

    Plasmodium falciparum (Pf) hypoxanthine-guanine phosphoribosyltransferase (HGPRT) is a potential therapeutic target. Compared to structurally homologous human enzymes, it has expanded substrate specificity. In this study, 9-deazapurines are used as in situ probes of the active sites of human and Pf HGPRTs. Through the use of these probes it is found that non-covalent interactions stabilise the pre-transition state of the HGPRT-catalysed reaction. Vibrational spectra reveal that the bound substrates are extensively distorted, the carbonyl bond of nucleobase moiety is weakened and the substrate is destabilised along the reaction coordinate. Raman shifts of the human and Pf enzymes are used to quantify the differing degrees of hydrogen bonding in the homologues. A decreased Raman cross-section in enzyme-bound 9-deazaguanine (9DAG) shows that the phenylalanine residue (Phe186 in human and Phe197 in Pf) of HGPRT stacks with the nucleobase. Differential loss of the Raman cross-section suggests that the active site is more compact in human HGPRT as compared to the Pf enzyme, and is more so in the phosphoribosyl pyrophosphate (PRPP) complex 9DAG-PRPP-HGPRT than in 9-deazahypoxanthine (9DAH)-PRPP-HGPRT. PMID:25944719

  5. Lipid-Induced Insulin Resistance in Skeletal Muscle: The Chase for the Culprit Goes from Total Intramuscular Fat to Lipid Intermediates, and Finally to Species of Lipid Intermediates.

    PubMed

    Kitessa, Soressa M; Abeywardena, Mahinda Y

    2016-01-01

    The skeletal muscle is the largest organ in the body. It plays a particularly pivotal role in glucose homeostasis, as it can account for up to 40% of the body and for up to 80%-90% of insulin-stimulated glucose disposal. Hence, insulin resistance (IR) in skeletal muscle has been a focus of much research and review. The fact that skeletal muscle IR precedes β-cell dysfunction makes it an ideal target for countering the diabetes epidemic. It is generally accepted that the accumulation of lipids in the skeletal muscle, due to dietary lipid oversupply, is closely linked with IR. Our understanding of this link between intramyocellular lipids (IMCL) and glycemic control has changed over the years. Initially, skeletal muscle IR was related to total IMCL. The inconsistencies in this explanation led to the discovery that particular lipid intermediates are more important than total IMCL. The two most commonly cited lipid intermediates for causing skeletal muscle IR are ceramides and diacylglycerol (DAG) in IMCL. Still, not all cases of IR and dysfunction in glycemic control have shown an increase in either or both of these lipids. In this review, we will summarise the latest research results that, using the lipidomics approach, have elucidated DAG and ceramide species that are involved in skeletal muscle IR in animal models and human subjects. PMID:27483311

  6. Properties and oxidative stabilities of enzymatically interesterified chicken fat and sunflower oil blend.

    PubMed

    Kostecka, Magdalena; Kowalska, Dorota; Kozłowska, Mariola; Kowalski, Boleslaw

    2013-01-01

    Chicken fat and sunflower oil 2:3 m/m blend was enzymatically interesterified at 60°C with and without microwaves assistance. As the catalyst a commercial preparation of the immobilized lipase from Rhizomucor miehei (Lipozyme RM IM) containing 2% of water was used, and the catalyst load was 8% in each case. The starting mixture and the interesterified products were separated by column chromatography into pure triacylglycerols fraction (TAG) and a non-triacylglycerol fraction, which contained free fatty acids (FFA), mono- and diacylglycerols (MAG and DAG). The oxidative stabilities (OS) of fats studied and TAG derived from them were assessed by Rancimat at 100°C and by Pressure Differential Scanning Calorimetry (PDSC) under oxygen at 110-140°C. Interesterification reduced the OS of chicken fat and sunflower oil blend. The main factors influenced on the OS of fats studied were concentrations of tocopherols and presence of FFA, MAG- and DAG. The structures of TAGs were of minor importance. From the resulting PDSC exotherms their times to reach the onset (τon) and peak maximum (τmax) were measured and used for calculations of parameters of the Arrhenius type kinetics for thermaloxidative decomposition of fats studied. PMID:24200936

  7. Handling Trajectory Uncertainties for Airborne Conflict Management

    NASA Technical Reports Server (NTRS)

    Barhydt, Richard; Doble, Nathan A.; Karr, David; Palmer, Michael T.

    2005-01-01

    Airborne conflict management is an enabling capability for NASA's Distributed Air-Ground Traffic Management (DAG-TM) concept. DAGTM has the goal of significantly increasing capacity within the National Airspace System, while maintaining or improving safety. Under DAG-TM, autonomous aircraft maintain separation from each other and from managed aircraft unequipped for autonomous flight. NASA Langley Research Center has developed the Autonomous Operations Planner (AOP), an onboard decision support system that provides airborne conflict management (ACM) and strategic flight planning support for autonomous aircraft pilots. The AOP performs conflict detection, prevention, and resolution from nearby traffic aircraft and area hazards. Traffic trajectory information is assumed to be provided by Automatic Dependent Surveillance Broadcast (ADS-B). Reliable trajectory prediction is a key capability for providing effective ACM functions. Trajectory uncertainties due to environmental effects, differences in aircraft systems and performance, and unknown intent information lead to prediction errors that can adversely affect AOP performance. To accommodate these uncertainties, the AOP has been enhanced to create cross-track, vertical, and along-track buffers along the predicted trajectories of both ownship and traffic aircraft. These buffers will be structured based on prediction errors noted from previous simulations such as a recent Joint Experiment between NASA Ames and Langley Research Centers and from other outside studies. Currently defined ADS-B parameters related to navigation capability, trajectory type, and path conformance will be used to support the algorithms that generate the buffers.

  8. Age-related changes in retinoic, docosahexaenoic and arachidonic acid modulation in nuclear lipid metabolism.

    PubMed

    Gaveglio, Virginia L; Pascual, Ana C; Giusto, Norma M; Pasquaré, Susana J

    2016-08-15

    The aim of this work was to study how age-related changes could modify several enzymatic activities that regulate lipid mediator levels in nuclei from rat cerebellum and how these changes are modulated by all-trans retinoic acid (RA), docosahexaenoic acid (DHA) and arachidonic acid (AA). The higher phosphatidate phosphohydrolase activity and lower diacylglycerol lipase (DAGL) activity observed in aged animals compared with adults could augment diacylglycerol (DAG) availability in the former. Additionally, monoacylglycerol (MAG) availability could be high due to an increase in lysophosphatidate phosphohydrolase (LPAPase) activity and a decrease in monocylglycerol lipase activity. Interestingly, RA, DHA and AA were observed to modulate these enzymatic activities and this modulation was found to change in aged rats. In adult nuclei, whereas RA led to high DAG and MAG production through inhibition of their hydrolytic enzymes, DHA and AA promoted high MAG production by LPAPase and DAGL stimulation. In contrast, in aged nuclei RA caused high MAG generation whereas DHA and AA diminished it through LPAPase activity modulation. These results demonstrate that aging promotes a different nuclear lipid metabolism as well as a different type of non-genomic regulation by RA, DHA and AA, which could be involved in nuclear signaling events. PMID:27355428

  9. Plant phospholipases D and C and their diverse functions in stress responses.

    PubMed

    Hong, Yueyun; Zhao, Jian; Guo, Liang; Kim, Sang-Chul; Deng, Xianjun; Wang, Geliang; Zhang, Gaoyang; Li, Maoyin; Wang, Xuemin

    2016-04-01

    Phospholipases D (PLD) and C (PLC) hydrolyze the phosphodiesteric linkages of the head group of membrane phospholipids. PLDs and PLCs in plants occur in different forms: the calcium-dependent phospholipid binding domain-containing PLDs (C2-PLDs), the plekstrin homology and phox homology domain-containing PLDs (PX/PH-PLDs), phosphoinositide-specific PLC (PI-PLC), and non-specific PLC (NPC). They differ in structures, substrate selectivities, cofactor requirements, and/or reaction conditions. These enzymes and their reaction products, such as phosphatidic acid (PA), diacylglycerol (DAG), and inositol polyphosphates, play important, multifaceted roles in plant response to abiotic and biotic stresses. Here, we review biochemical properties, cellular effects, and physiological functions of PLDs and PLCs, particularly in the context of their roles in stress response along with advances made on the role of PA and DAG in cell signaling in plants. The mechanism of actions, including those common and distinguishable among different PLDs and PLCs, will also be discussed. PMID:26783886

  10. Antidiabetic Effect and Mode of Action of Cytopiloyne

    PubMed Central

    Chang, Cicero Lee-Tian; Liu, Hsien-Yueh; Kuo, Tien-Fen; Hsu, Yi-Jou; Shen, Ming-Yi; Pan, Chien-Yuan

    2013-01-01

    Cytopiloyne was identified as a novel polyacetylenic compound. However, its antidiabetic properties are poorly understood. The aim of the present study was to investigate the anti-diabetic effect and mode of action of cytopiloyne on type 2 diabetes (T2D). We first evaluated the therapeutic effect of cytopiloyne on T2D in db/db mice. We found that one dose of cytopiloyne reduced postprandial glucose levels while increasing blood insulin levels. Accordingly, long-term treatment with cytopiloyne reduced postprandial blood glucose levels, increased blood insulin, improved glucose tolerance, suppressed the level of glycosylated hemoglobin A1c (HbA1c), and protected pancreatic islets in db/db mice. Next, we studied the anti-diabetic mechanism of action of cytopiloyne. We showed that cytopiloyne failed to decrease blood glucose in streptozocin- (STZ-)treated mice whose β cells were already destroyed. Additionally, cytopiloyne dose dependently increased insulin secretion and expression in β cells. The increase of insulin secretion/expression of cytopiloyne was regulated by protein kinase Cα (PKCα) and its activators, calcium, and diacylglycerol (DAG). Overall, our data suggest that cytopiloyne treats T2D via regulation of insulin production involving the calcium/DAG/PKCα cascade in β cells. These data thus identify the molecular mechanism of action of cytopiloyne and prove its therapeutic potential in T2D. PMID:23573144

  11. SU-E-J-142: Performance Study of Automatic Image-Segmentation Algorithms in Motion Tracking Via MR-IGRT

    SciTech Connect

    Feng, Y; Olsen, J.; Parikh, P.; Noel, C; Wooten, H; Du, D; Mutic, S; Hu, Y; Kawrakow, I; Dempsey, J

    2014-06-01

    , different filtering methods and their influences on the segmentation results. Parag Parikh receives research grant from ViewRay. Sasa Mutic has consulting and research agreements with ViewRay. Yanle Hu receives travel reimbursement from ViewRay. Iwan Kawrakow and James Dempsey are ViewRay employees.

  12. ⁵³Mn-⁵³Cr and ²⁶Al-²⁶Mg ages of a feldspathic lithology in polymict ureilites

    SciTech Connect

    Goodrich, Cyrena Anne; Hutcheon, Ian D.; Kita, Noriko T.; Huss, Gary R.; Cohen, Barbara Anne; Keil, Klaus

    2010-07-01

    We report 53Mn–53Cr and 26Al–26Mg isotopic data, obtained by in-situ SIMS analysis, for feldspathic clasts in polymict ureilites DaG 319 and DaG 165. The analyzed clasts belong to the “albitic lithology,” the most abundant population of indigenous feldspathic materials in polymict ureilites, and are highly fractionated igneous assemblages of albitic plagioclase, Fe-rich pyroxenes, phosphates, ilmenite, silica, and Fe(Mn, K, P, Ti)-enriched glass. Glass in DaG 165 clast 19 has extremely high and variable 55Mn/52Cr ratios (500–58,000) and shows correlated 53Cr excesses up to ~ 1500‰, clearly indicating the presence of live 53Mn at the time of formation. The slope of the well-correlated isochron defined by glass and pyroxenes from this clast corresponds to (53Mn/55Mn) = (2.84 ± 0.10) × 10-6 (2σ). Data for less 55Mn/52Cr-enriched glasses from DaG 319 clast B1, as well as phosphates from several other clasts, are consistent with this isochron. The 53Mn/55Mn ratio obtained from the isochron implies that these clasts are 0.70 ± 0.18 Ma younger than the D'Orbigny angrite, corresponding to the absolute age of 4563.72 ± 0.22 Ma. Plagioclase in DaG 319 clast B1 has a fairly constant 27Al/24Mg ratio of ~ 900 and shows resolvable 26Mg excesses of ~ 2‰. The slope of the isochron defined by pyroxene and plagioclase in this clast is (3.0 ± 1.1) × 10-7 (2σ), corresponding to a time difference of 5.4 (-0.3/+0.5) Ma after CAI (assuming the canonical initial 26Al/27Al ratio of 5 × 10-5) and an age 0.5 (-0.3/+0.5) Ma younger than D'Orbigny. Its absolute age (relative to D'Orbigny) is 4563.9 (+ 0.4/-0.5) Ma, in agreement with the 53Mn–53Cr age from clast 19. These data provide the first high

  13. RasGRP1 overexpression in T-ALL increases basal nucleotide exchange on Ras rendering the Ras/PI3K/Akt pathway responsive to protumorigenic cytokines.

    PubMed

    Ksionda, O; Melton, A A; Bache, J; Tenhagen, M; Bakker, J; Harvey, R; Winter, S S; Rubio, I; Roose, J P

    2016-07-14

    Ras GTPases are activated by RasGEFs and inactivated by RasGAPs, which stimulate the hydrolysis of RasGTP to inactive RasGDP. GTPase-impairing somatic mutations in RAS genes, such as KRAS(G12D), are among the most common oncogenic events in metastatic cancer. A different type of cancer Ras signal, driven by overexpression of the RasGEF RasGRP1 (Ras guanine nucleotide-releasing protein 1), was recently implicated in pediatric T-cell acute lymphoblastic leukemia (T-ALL) patients and murine models, in which RasGRP1 T-ALLs expand in response to treatment with interleukins (ILs) 2, 7 and 9. Here, we demonstrate that IL-2/7/9 stimulation activates Erk and Akt pathways downstream of Ras in RasGRP1 T-ALL but not in normal thymocytes. In normal lymphocytes, RasGRP1 is recruited to the membrane by diacylglycerol (DAG) in a phospholipase C-γ (PLCγ)-dependent manner. Surprisingly, we find that leukemic RasGRP1-triggered Ras-Akt signals do not depend on acute activation of PLCγ to generate DAG but rely on baseline DAG levels instead. In agreement, using three distinct assays that measure different aspects of the RasGTP/GDP cycle, we established that overexpression of RasGRP1 in T-ALLs results in a constitutively high GTP-loading rate of Ras, which is constantly counterbalanced by hydrolysis of RasGTP. KRAS(G12D) T-ALLs do not show constitutive GTP loading of Ras. Thus, we reveal an entirely novel type of leukemogenic Ras signals that is based on a RasGRP1-driven increased in flux through the RasGTP/GDP cycle, which is mechanistically very different from KRAS(G12D) signals. Our studies highlight the dynamic balance between RasGEF and RasGAP in these T-ALLs and put forth a new model in which IL-2/7/9 decrease RasGAP activity. PMID:26549032

  14. Platelet-derived growth factor stimulates protein kinase D through the activation of phospholipase Cgamma and protein kinase C.

    PubMed

    Van Lint, J; Ni, Y; Valius, M; Merlevede, W; Vandenheede, J R

    1998-03-20

    Platelet-derived growth factor (PDGF) stimulates protein kinase D (PKD) in a time- and dose-dependent manner. We have used a series of PDGF receptor mutants that display a selective impairment of the binding of SH2-containing proteins (GTPase-activating protein, SHP-2, phospholipase Cgamma (PLCgamma), or phosphatidylinositol 3'-kinase (PI3K)) to show that Tyr-1021, the PLCgamma-binding site, is essential for PKD stimulation by PDGF in A431 cells. We next investigated whether any one of these four binding sites could mediate PKD activation in the absence of the other three sites. F5, a receptor mutant that lacks all four binding sites for GTPase-activating protein, PLCgamma, PI3K, and SHP-2, fails to activate PKD. A panel of single add-back mutants was used to investigate if any one of these four sites could restore signaling to PKD. Of the four sites, only the PLCgamma+ single add-back receptor restored PDGF-mediated activation of PKD, and only this add-back receptor produced diacylglycerol (DAG) in a PDGF-dependent manner. 1,2-Dioctanoyl-sn-glycerol, a membrane-permeant DAG analog, was found to be sufficient for activation of PKD. Taken together, these data indicate that PLCgamma activation is not only necessary, but also sufficient to mediate PDGF-induced PKD activation. Although the presence of a pleckstrin homology domain makes PKD a potential PI3K target, PKD was not stimulated by selective PI3K activation, and wortmannin, an inhibitor of PI3K, did not inhibit PDGF signaling to PKD. The activation of PKD by DAG or by the wild-type and PLCgamma+ add-back PDGF receptors was inhibited by GF109203X, suggesting a role for protein kinase C in the stimulation of PKD by PDGF. PDGF induced a time-dependent phosphorylation of PKD that closely correlated with activation. The PDGF-induced activation and phosphorylation of PKD were reversed by in vitro incubation of PKD with protein phosphatase 1 or 2A, indicating that PDGF signaling to PKD involves the Ser

  15. Spatial and temporal variability of biomarkers and microbial diversity reveal metabolic and community flexibility in Streamer Biofilm Communities in the Lower Geyser Basin, Yellowstone National Park.

    PubMed

    Schubotz, F; Meyer-Dombard, D R; Bradley, A S; Fredricks, H F; Hinrichs, K-U; Shock, E L; Summons, R E

    2013-11-01

    Detailed analysis of 16S rRNA and intact polar lipids (IPLs) from streamer biofilm communities (SBCs), collected from geochemically similar hot springs in the Lower Geyser Basin, Yellowstone National Park, shows good agreement and affirm that IPLs can be used as reliable markers for the microbial constituents of SBCs. Uncultured Crenarchaea are prominent in SBS, and their IPLs contain both glycosidic and mixed glyco-phospho head groups with tetraether cores, having 0-4 rings. Archaeal IPL contributions increase with increasing temperature and comprise up to one-fourth of the total IPL inventory at >84 °C. At elevated temperatures, bacterial IPLs contain abundant glycosidic glycerol diether lipids. Diether and diacylglycerol (DAG) lipids with aminopentanetetrol and phosphatidylinositol head groups were identified as lipids diagnostic of Aquificales, while DAG glycolipids and glyco-phospholipids containing N-acetylgycosamine as head group were assigned to members of the Thermales. With decreasing temperature and concomitant changes in water chemistry, IPLs typical of phototrophic bacteria, such as mono-, diglycosyl, and sulfoquinovosyl DAG, which are specific for cyanobacteria, increase in abundance, consistent with genomic data from the same samples. Compound-specific stable carbon isotope analysis of IPL breakdown products reveals a large isotopic diversity among SBCs in different hot springs. At two of the hot springs, 'Bison Pool' and Flat Cone, lipids derived from Aquificales are enriched in (13) C relative to biomass and approach values close to dissolved inorganic carbon (DIC) (approximately 0‰), consistent with fractionation during autotrophic carbon fixation via the reversed tricarboxylic acid pathway. At a third site, Octopus Spring, the same Aquificales-diagnostic lipids are 10‰ depleted relative to biomass and resemble stable carbon isotope values of dissolved organic carbon (DOC), indicative of heterotrophy. Other bacterial and archaeal lipids show

  16. Reduced Skeletal Muscle Oxidative Capacity and Elevated Ceramide but not Diacylglycerol Content in Severe Obesity

    PubMed Central

    Coen, P.M.; Hames, K.C.; Leachman, E.M.; DeLany, J.P.; Ritov, V.B.; Menshikova, E.V.; Dubé, J.J.; Stefanovic-Racic, M.; Toledo, F.G.S.; Goodpaster, B.H.

    2014-01-01

    Objective The link between a reduced capacity for skeletal muscle mitochondrial fatty acid oxidation (FAO) and lipotoxicity in human insulin resistance has been the subject of intense debate. The objective of this study was to investigate whether reduced FAO is associated with elevated acyl CoA, ceramide, and diacylglycerol (DAG) in severely obese insulin resistant subjects. Design and Methods Muscle biopsies were conducted in lean (L, 22.6 ± 0.5 kg/m2, n = 8), Class I (CI, 32.1 ± 0.4 kg/m2, n = 7) and Class II&III obese (CII&III, 45.6 ± 1.1 kg/m2, n = 15) women for acyl CoA, sphingolipid and DAG profiling. Intramyocellular triglyceride (IMTG) content was determined by histology. FAO was assessed by incubating muscle homogenates with [1–14C]palmitate and measuring 14CO2 production. Cardiolipin content was quantified as an index of mitochondrial content. Lipid metabolism proteins, DGAT1, PLIN5, and PNPLA2 were quantified in biopsy samples by western blot. Results CII&III were more insulin resistant (HOMA-IR: 4.5 ± 0.5 vs. 1.1 ± 0.1, P < 0.001), and had lower FAO (~58%, P = 0.007) and cardiolipin content (~31%, P = 0.013) compared to L. IMTG was elevated in CI (P = 0.04) and CII&III (P = 0.04) compared to L. Sphingolipid content was higher in CII&III compared to L (13.6 ± 1.1 vs. 10.3 ± 0.5 pmol/mg, P = 0.031) whereas DAG content was not different among groups. DGAT1 was elevated in CII&III, and PLIN5 was elevated in CI compared to L. Conclusions Severe obesity is associated with reduced muscle oxidative capacity and occurs concomitantly with elevated IMTG, ceramide and insulin resistance. PMID:23512750

  17. Role of transient receptor potential canonical 6 (TRPC6) in non-transferrin-bound iron uptake in neuronal phenotype PC12 cells.

    PubMed Central

    Mwanjewe, James; Grover, Ashok K

    2004-01-01

    Cells take up transferrin-bound iron or NTBI (non-transferrin-bound iron). After treatment with NGF (nerve growth factor), PC12 cells exhibited a neuronal phenotype and an increase in the NTBI uptake (55Fe2+ or 55Fe3+). We loaded the cells with the dye calcein, whose fluorescence increases in the presence of Ca2+ but is quenched with Fe2+ or Fe3+. When examined using calcein fluorescence or radioactive iron, DAG (diacylglycerol)-stimulated NTBI entry was more in NGF-treated PC12 cells compared with untreated cells. All experiments were performed at 1.5 mM extracellular Ca2+. Nramp2 (natural-resistance-associated macrophage protein 2) mRNA expression did not change after the NGF treatment. Expression of the bivalent cation entry protein TRPC6 (transient receptor potential canonical 6) was detected only in the NGF-treated cells. To verify that increased NTBI uptake depended on TRPC6, we examined whether transfecting HEK-293 (human embryonic kidney 293) cells with TRPC6 also increased the NTBI (55Fe) uptake. We also cotransfected HEK-293 cells with two plasmids, one expressing TRPC6 and the other expressing the fluorescent protein DsRED2 to identify the transfected cells. Challenging the calcein-loaded HEK-293 cells (which intrinsically express the a1-adrenergic receptors) with phenylephrine or a cell-permeant DAG increased the fluorescence signal more rapidly in transfected cells compared with untransfected cells. However, when iron (Fe2+ and Fe3+) was added before adding phenylephrine or DAG, the fluorescence intensity decreased more rapidly in transfected cells compared with untransfected cells, thereby indicating a greater stimulation of the NTBI uptake in cells expressing TRPC6. We postulate that the increase in the NTBI entry into neuronal PC12 cells is through TRPC6, a pathway that is unique since it is receptor-stimulated. Since neuronal cells express TRPC6, this pathway may have a role in neurotoxicity. PMID:14640978

  18. Identification and physiological characterization of phosphatidic acid phosphatase enzymes involved in triacylglycerol biosynthesis in Streptomyces coelicolor

    PubMed Central

    2013-01-01

    Background Phosphatidic acid phosphatase (PAP, EC 3.1.3.4) catalyzes the dephosphorylation of phosphatidate yielding diacylglycerol (DAG), the lipid precursor for triacylglycerol (TAG) biosynthesis. Despite the importance of PAP activity in TAG producing bacteria, studies to establish its role in lipid metabolism have been so far restricted only to eukaryotes. Considering the increasing interest of bacterial TAG as a potential source of raw material for biofuel production, we have focused our studies on the identification and physiological characterization of the putative PAP present in the TAG producing bacterium Streptomyces coelicolor. Results We have identified two S. coelicolor genes, named lppα (SCO1102) and lppβ (SCO1753), encoding for functional PAP proteins. Both enzymes mediate, at least in part, the formation of DAG for neutral lipid biosynthesis. Heterologous expression of lppα and lppβ genes in E. coli resulted in enhanced PAP activity in the membrane fractions of the recombinant strains and concomitantly in higher levels of DAG. In addition, the expression of these genes in yeast complemented the temperature-sensitive growth phenotype of the PAP deficient strain GHY58 (dpp1lpp1pah1). In S. coelicolor, disruption of either lppα or lppβ had no effect on TAG accumulation; however, the simultaneous mutation of both genes provoked a drastic reduction in de novo TAG biosynthesis as well as in total TAG content. Consistently, overexpression of Lppα and Lppβ in the wild type strain of S. coelicolor led to a significant increase in TAG production. Conclusions The present study describes the identification of PAP enzymes in bacteria and provides further insights on the genetic basis for prokaryotic oiliness. Furthermore, this finding completes the whole set of enzymes required for de novo TAG biosynthesis pathway in S. coelicolor. Remarkably, the overexpression of these PAPs in Streptomyces bacteria contributes to a higher productivity of this single

  19. Protein Kinase D Enzymes as Regulators of EMT and Cancer Cell Invasion.

    PubMed

    Durand, Nisha; Borges, Sahra; Storz, Peter

    2016-01-01

    The Protein Kinase D (PKD) isoforms PKD1, PKD2, and PKD3 are effectors of the novel Protein Kinase Cs (nPKCs) and diacylglycerol (DAG). PKDs impact diverse biological processes like protein transport, cell migration, proliferation, epithelial to mesenchymal transition (EMT) and apoptosis. PKDs however, have distinct effects on these functions. While PKD1 blocks EMT and cell migration, PKD2 and PKD3 tend to drive both processes. Given the importance of EMT and cell migration to the initiation and progression of various malignancies, abnormal expression of PKDs has been reported in multiple types of cancers, including breast, pancreatic and prostate cancer. In this review, we discuss how EMT and cell migration are regulated by PKD isoforms and the significance of this regulation in the context of cancer development. PMID:26848698

  20. A direct method for regiospecific analysis of TAG using alpha-MAG.

    PubMed

    Turon, F; Bachain, P; Caro, Y; Pina, M; Graille, J

    2002-08-01

    An analytical procedure was developed for regiodistribution analysis of TAG using alpha-MAG prepared by an ethyl magnesium bromide deacylation. In the present communication, the deacylation procedure is shown to lead to representative alpha-MAG, allowing the composition of the native TAG in the alpha-position to be determined directly. The composition in the beta-position can then be estimated from the composition of the alpha-MAG and TAG according to the formula 3 x TAG - 2 x alpha-MAG. The estimates are superior to those obtained using the alpha,beta-DAG and Brockerhoff calculations as they come closer to the theoretical value and have smaller SD. The present procedure, first demonstrated on a synthetic TAG, was then successfully applied to the analysis of borage oil, milkfat, and tuna oil. PMID:12371754

  1. Rapid and profound rewiring of brain lipid signaling networks by acute diacylglycerol lipase inhibition.

    PubMed

    Ogasawara, Daisuke; Deng, Hui; Viader, Andreu; Baggelaar, Marc P; Breman, Arjen; den Dulk, Hans; van den Nieuwendijk, Adriann M C H; Soethoudt, Marjolein; van der Wel, Tom; Zhou, Juan; Overkleeft, Herman S; Sanchez-Alavez, Manuel; Mo, Simone; Nguyen, William; Conti, Bruno; Liu, Xiaojie; Chen, Yao; Liu, Qing-Song; Cravatt, Benjamin F; van der Stelt, Mario

    2016-01-01

    Diacylglycerol lipases (DAGLα and DAGLβ) convert diacylglycerol to the endocannabinoid 2-arachidonoylglycerol. Our understanding of DAGL function has been hindered by a lack of chemical probes that can perturb these enzymes in vivo. Here, we report a set of centrally active DAGL inhibitors and a structurally related control probe and their use, in combination with chemical proteomics and lipidomics, to determine the impact of acute DAGL blockade on brain lipid networks in mice. Within 2 h, DAGL inhibition produced a striking reorganization of bioactive lipids, including elevations in DAGs and reductions in endocannabinoids and eicosanoids. We also found that DAGLα is a short half-life protein, and the inactivation of DAGLs disrupts cannabinoid receptor-dependent synaptic plasticity and impairs neuroinflammatory responses, including lipopolysaccharide-induced anapyrexia. These findings illuminate the highly interconnected and dynamic nature of lipid signaling pathways in the brain and the central role that DAGL enzymes play in regulating this network. PMID:26668358

  2. FIFE-Jobsub: a grid submission system for intensity frontier experiments at Fermilab

    SciTech Connect

    Box, Dennis

    2014-01-01

    The Fermilab Intensity Frontier Experiments use an integrated submission system known as FIFE-jobsub, part of the FIFE (Fabric for Frontier Experiments) initiative, to submit batch jobs to the Open Science Grid. FIFE-jobsub eases the burden on experimenters by integrating data transfer and site selection details in an easy to use and well-documented format. FIFE-jobsub automates tedious details of maintaining grid proxies for the lifetime of the grid job. Data transfer is handled using the Intensity Frontier Data Handling Client (IFDHC) [1] tool suite, which facilitates selecting the appropriate data transfer method from many possibilities while protecting shared resources from overload. Chaining of job dependencies into Directed Acyclic Graphs (Condor DAGS) is well supported and made easier through the use of input flags and parameters.

  3. Mutations in the midway gene disrupt a Drosophila acyl coenzyme A: diacylglycerol acyltransferase.

    PubMed Central

    Buszczak, Michael; Lu, Xiaohui; Segraves, William A; Chang, Ta Yuan; Cooley, Lynn

    2002-01-01

    During Drosophila oogenesis, defective or unwanted egg chambers are eliminated during mid-oogenesis by programmed cell death. In addition, final cytoplasm transport from nurse cells to the oocyte depends upon apoptosis of the nurse cells. To study the regulation of germline apoptosis, we analyzed the midway mutant, in which egg chambers undergo premature nurse cell death and degeneration. The midway gene encodes a protein similar to mammalian acyl coenzyme A: diacylglycerol acyltransferase (DGAT), which converts diacylglycerol (DAG) into triacylglycerol (TAG). midway mutant egg chambers contain severely reduced levels of neutral lipids in the germline. Expression of midway in insect cells results in high levels of DGAT activity in vitro. These results show that midway encodes a functional DGAT and that changes in acylglycerol lipid metabolism disrupt normal egg chamber development in Drosophila. PMID:11973306

  4. IceProd 2: A Next Generation Data Analysis Framework for the IceCube Neutrino Observatory

    NASA Astrophysics Data System (ADS)

    Schultz, D.

    2015-12-01

    We describe the overall structure and new features of the second generation of IceProd, a data processing and management framework. IceProd was developed by the IceCube Neutrino Observatory for processing of Monte Carlo simulations, detector data, and analysis levels. It runs as a separate layer on top of grid and batch systems. This is accomplished by a set of daemons which process job workflow, maintaining configuration and status information on the job before, during, and after processing. IceProd can also manage complex workflow DAGs across distributed computing grids in order to optimize usage of resources. IceProd is designed to be very light-weight; it runs as a python application fully in user space and can be set up easily. For the initial completion of this second version of IceProd, improvements have been made to increase security, reliability, scalability, and ease of use.

  5. Functionalization of Repetitive Polypeptides for Molecular Interconnect Applications

    NASA Astrophysics Data System (ADS)

    Carlsen, A.; Rana, N.; Kossow, C.; Cheng, D.; Wells, C.; Bousman, K.; Ngo, S.; Higashiya, S.; Welch, J.; Raynolds, J.; Dunn, K.; Eisenbraun, E.; Geer, R.; Kaloyeros, A.

    2004-03-01

    This study focuses on the functionalization of a genetically engineered molecule consisting of a repetitive polypeptide sequence [(GA)_3GY(GA)_3GE(GA)_3GH(GA)_3GK where G=glycine, A=alanine, Y=tyrosine, E=glutamic acid, H=histidine, and K=lysine] drawn into a β -pleated sheet formation.^ The polyhistidinyl tracts decorating this β -sheet ``scaffolding'' act as a repeating array of functional moieties for the attachment of metallic ions to facilitate charge transport. A second functionalization approach investigated utilizes the electrostatic interaction between anionic portions of the molecule and cationic Au nanoparticles. The facility of these approaches was demonstrated using scanning probe microscopy, circular dichroism, and Raman spectroscopy. ^dag S. Higashiya, S. Ngo, K. Bousman, J. Welch, N. Rana, A. Carlsen, E. Eisenbraun, R. Geer, A. Kaloyeros. Polym Mat Sci Engin. 89, 466-7 (2003).

  6. Glycerolipid biosynthesis in isolated pea root plastids

    SciTech Connect

    Xue, Lingru; Sparace, S.A. )

    1990-05-01

    Plastids have been isolated from germinating pea (Pisum sativum L.) roots by differential centrifugation and purified on Percoll gradients. Marker enzymes (NADPH: cytochrome c reductase, fumarase and fatty acid synthesis) indicate that greater than 50% of the plastids are recovered essentially free from mitochondrial and endoplasmic reticulum contamination. Fatty acids synthesized from ({sup 14}C)acetate by Percoll-purified plastids are primarily 16:0, 16:1 and 18:1. ({sup 14}C)Acetate-labelled fatty acids and ({sup 14}C)glycerol-3-phosphate are both readily incorporated into glycerolipid. Approximately 12% of the total activity for glycerolipid biosynthesis from glycerol-3-phosphate is recovered in the purified plastid fraction. Glycerolipids synthesized from these precursors are primarily TAG, DAG, PE, PG, PC, PI and PA. Acyl-CoA's also accumulate when acetate is the precursor.

  7. Protein Kinase D Enzymes as Regulators of EMT and Cancer Cell Invasion

    PubMed Central

    Durand, Nisha; Borges, Sahra; Storz, Peter

    2016-01-01

    The Protein Kinase D (PKD) isoforms PKD1, PKD2, and PKD3 are effectors of the novel Protein Kinase Cs (nPKCs) and diacylglycerol (DAG). PKDs impact diverse biological processes like protein transport, cell migration, proliferation, epithelial to mesenchymal transition (EMT) and apoptosis. PKDs however, have distinct effects on these functions. While PKD1 blocks EMT and cell migration, PKD2 and PKD3 tend to drive both processes. Given the importance of EMT and cell migration to the initiation and progression of various malignancies, abnormal expression of PKDs has been reported in multiple types of cancers, including breast, pancreatic and prostate cancer. In this review, we discuss how EMT and cell migration are regulated by PKD isoforms and the significance of this regulation in the context of cancer development. PMID:26848698

  8. Studio delle popolazioni di blazar ad alte energie

    NASA Astrophysics Data System (ADS)

    Gasparrini, D.; Giommi, P.

    2008-10-01

    Utilizzando la grande quantita' di dati sui blazar disponibile ad oggi, e' possibile costruire una distribuzione spettrale di energia (SED) media per ciascuna classe di oggetti. Basandosi su un modello SSC, e' inoltre possibile stimare il comportamento di questi oggetti nella banda di energia dei raggi gamma partendo dalle informazioni ottenute da osservazioni in altre bande di energia quali il radio e l'X. Utilizzando le simulazioni sara` possibile prevedere un eventuale cambio di popolazione dominante ad alte energie(oltre 1GeV) rispetto a quanto avviene in banda radio dove domina la popolazione dei FSRQ rispetto ai BL Lac e cercare un'eventuale conferma dagli esperimenti AGILE e GLAST.

  9. Lipase catalyzed synthesis of neutral glycerides rich in micronutrients from rice bran oil fatty acid distillate.

    PubMed

    Nandi, Sumit; Gangopadhyay, Sarbani; Ghosh, Santinath

    2008-01-01

    Neutral glycerides with micronutrients like sterols, tocopherols and squalene may be prepared from cheap raw material like rice bran oil fatty acid distillate (RBO FAD). RBO FAD is an important byproduct of vegetable oil refining industries in the physical refining process. Glycerides like triacylglycerols (TAG), diacylglycerols (DAG) and monoacylglycerols (MAG) containing significant amounts of unsaponifiable matter like sterols, tocopherols and hydrocarbons (mainly squalene) may certainly be considered as novel functional food ingredients. Fatty acids present in RBO FAD were esterified with glycerol of varying amount (1:0.33, 1:0.5, 1:1 and 1:1.5 of FAD : glycerol ratio) for 8 h using non-specific enzyme NS 40013 (Candida antartica). After esterification the product mixture containing mono, di- and triglycerides was purified by molecular distillation to remove excess free fatty acids and also other volatile undesirable components. The purified product containing sterols, tocopherols and squalene can be utilized in various food formulations. PMID:18838832

  10. Prime Numbers, Quantum Field Theory and the Goldbach Conjecture

    NASA Astrophysics Data System (ADS)

    Sanchis-Lozano, Miguel-Angel; Barbero G., J. Fernando; Navarro-Salas, José

    2012-09-01

    Motivated by the Goldbach conjecture in number theory and the Abelian bosonization mechanism on a cylindrical two-dimensional space-time, we study the reconstruction of a real scalar field as a product of two real fermion (so-called prime) fields whose Fourier expansion exclusively contains prime modes. We undertake the canonical quantization of such prime fields and construct the corresponding Fock space by introducing creation operators bp\\dag — labeled by prime numbers p — acting on the vacuum. The analysis of our model, based on the standard rules of quantum field theory and the assumption of the Riemann hypothesis, allows us to prove that the theory is not renormalizable. We also comment on the potential consequences of this result concerning the validity or breakdown of the Goldbach conjecture for large integer numbers.

  11. Instantaneous characterization of vegetable oils via TAG and FFA profiles by easy ambient sonic-spray ionization mass spectrometry.

    PubMed

    Simas, Rosineide C; Catharino, Rodrigo R; Cunha, Ildenize B S; Cabral, Elaine C; Barrera-Arellano, Daniel; Eberlin, Marcos N; Alberici, Rosana M

    2010-04-01

    A fast and reliable method is presented for the analysis of vegetable oils. Easy ambient sonic-spray ionization mass spectrometry (EASI-MS) is shown to efficiently desorb and ionize the main oil constituents from an inert surface under ambient conditions and to provide comprehensive triacylglyceride (TAG) and free fatty acid (FFA) profiles detected mainly as either [TAG + Na](+) or [FFA-H](-) ions. EASI(+/-)-MS analysis is simple, easily implemented, requires just a tiny droplet of the oil and is performed without any pre-separation or chemical manipulation. It also causes no fragmentation of TAG ions hence diacylglyceride (DAG) and monoacylglyceride (MAG) profiles and contents can also be measured. The EASI(+/-)-MS profiles of TAG and FFA permit authentication and quality control and can be used, for instance, to access levels of adulteration, acidity, oxidation or hydrolysis of vegetable oils in general. PMID:20349539

  12. Analysis of the Lipidome of Xenografts Using MALDI-IMS and UHPLC-ESI-QTOF

    NASA Astrophysics Data System (ADS)

    Fernández, Roberto; Lage, Sergio; Abad-García, Beatriz; Barceló-Coblijn, Gwendolyn; Terés, Silvia; López, Daniel H.; Guardiola-Serrano, Francisca; Martín, M. Laura; Escribá, Pablo V.; Fernández, José A.

    2014-07-01

    Human tumor xenografts in immunodeficient mice are a very popular model to study the development of cancer and to test new drug candidates. Among the parameters analyzed are the variations in the lipid composition, as they are good indicators of changes in the cellular metabolism. Here, we present a study on the distribution of lipids in xenografts of NCI-H1975 human lung cancer cells, using MALDI imaging mass spectrometry and UHPLC-ESI-QTOF. The identification of lipids directly from the tissue by MALDI was aided by the comparison with identification using ESI ionization in lipid extracts from the same xenografts. Lipids belonging to PCs, PIs, SMs, DAG, TAG, PS, PA, and PG classes were identified and their distribution over the xenograft was determined. Three areas were identified in the xenograft, corresponding to cells in different metabolic stages and to a layer of adipose tissue that covers the xenograft.

  13. The last word

    NASA Astrophysics Data System (ADS)

    Daglis, Ioaanis A.

    This is the third in a series of essays on terms used in solar-terrestrial physics that are thought to be in need of clarification. Terms are identified and essays are commissioned by a committee chartered by Division II (Sun and Heliosphere) of the International Astronomical Union. Terminology Committee members include Ed Oliver (chair), Jean-Louis Bougeret, Hilary Cane, Takeo Kosugi, Sara Martin, Rainer Schwenn, Lidia van Driel-Gestelyi, and Joe Borovsky.Writers are asked to review the origins of a term and its current usage/misusage. The main point is to open a discussion and inform the community. We solicit letters to Eos on the following article by Ioannis Daglis on the term “magnetic storm,” which is certainly one of the most venerable names in solar-terrestrial physics. In addition, we welcome suggestions to any committee member on other terms to address in this forum.

  14. Canonical multi-valued input Reed-Muller trees and forms

    NASA Technical Reports Server (NTRS)

    Perkowski, M. A.; Johnson, P. D.

    1991-01-01

    There is recently an increased interest in logic synthesis using EXOR gates. The paper introduces the fundamental concept of Orthogonal Expansion, which generalizes the ring form of the Shannon expansion to the logic with multiple-valued (mv) inputs. Based on this concept we are able to define a family of canonical tree circuits. Such circuits can be considered for binary and multiple-valued input cases. They can be multi-level (trees and DAG's) or flattened to two-level AND-EXOR circuits. Input decoders similar to those used in Sum of Products (SOP) PLA's are used in realizations of multiple-valued input functions. In the case of the binary logic the family of flattened AND-EXOR circuits includes several forms discussed by Davio and Green. For the case of the logic with multiple-valued inputs, the family of the flattened mv AND-EXOR circuits includes three expansions known from literature and two new expansions.

  15. A Fast-Time Simulation Environment for Airborne Merging and Spacing Research

    NASA Technical Reports Server (NTRS)

    Bussink, Frank J. L.; Doble, Nathan A.; Barmore, Bryan E.; Singer, Sharon

    2005-01-01

    As part of NASA's Distributed Air/Ground Traffic Management (DAG-TM) effort, NASA Langley Research Center is developing concepts and algorithms for merging multiple aircraft arrival streams and precisely spacing aircraft over the runway threshold. An airborne tool has been created for this purpose, called Airborne Merging and Spacing for Terminal Arrivals (AMSTAR). To evaluate the performance of AMSTAR and complement human-in-the-loop experiments, a simulation environment has been developed that enables fast-time studies of AMSTAR operations. The environment is based on TMX, a multiple aircraft desktop simulation program created by the Netherlands National Aerospace Laboratory (NLR). This paper reviews the AMSTAR concept, discusses the integration of the AMSTAR algorithm into TMX and the enhancements added to TMX to support fast-time AMSTAR studies, and presents initial simulation results.

  16. Development of a Free-Flight Simulation Infrastructure

    NASA Technical Reports Server (NTRS)

    Miles, Eric S.; Wing, David J.; Davis, Paul C.

    1999-01-01

    In anticipation of a projected rise in demand for air transportation, NASA and the FAA are researching new air-traffic-management (ATM) concepts that fall under the paradigm known broadly as ":free flight". This paper documents the software development and engineering efforts in progress by Seagull Technology, to develop a free-flight simulation (FFSIM) that is intended to help NASA researchers test mature-state concepts for free flight, otherwise referred to in this paper as distributed air / ground traffic management (DAG TM). Under development is a distributed, human-in-the-loop simulation tool that is comprehensive in its consideration of current and envisioned communication, navigation and surveillance (CNS) components, and will allow evaluation of critical air and ground traffic management technologies from an overall systems perspective. The FFSIM infrastructure is designed to incorporate all three major components of the ATM triad: aircraft flight decks, air traffic control (ATC), and (eventually) airline operational control (AOC) centers.

  17. Tackling pollution by organic farming is capable of increasing fortified foods.

    PubMed

    Navarro-Aviñó, J P; Navarro, J J Fernández; Castro, V Vargas; Ripoll, I Ilzarbe; Sahuquillo, M J Márquez

    2016-06-01

    The global pollution stage is poisoning the biosphere and causing global temperatures to rise, necessitating a drastic change in the way man is dealing with nature. One change that may produce many beneficial effects on the biosphere and human health is the use of specific organic farming to produce food in a more integrated way in nature and to increase the capacity of man's own response. Despite many experts' opinion another way to deal with environmental contamination is possible: organic farming, which can increase man's ability to fortify foods. After more than 20 years working under this discipline, Bodegas Dagon is able to achieve the highest stilbenes concentrations (as resveratrol). Versus 14.3 mg/l, "Bodegas Dagón" wines contain resveratrol (HPLC and UV-spectroscopy) up to 1611.73 ± 72.66 mg/l, standing as world's potentially healthiest wine reported to date. PMID:26374929

  18. Comprehensive analysis of lipid composition in crude palm oil using multiple lipidomic approaches.

    PubMed

    Cheong, Wei Fun; Wenk, Markus R; Shui, Guanghou

    2014-05-20

    Palm oil is currently the leading edible oil consumed worldwide. Triacylglycerol (TAG) and diacylglycerol (DAG) are the dominant lipid classes in palm oil. Other lipid classes present in crude palm oil, such as phospholipids and galactolipids, are very low in abundance. These low-abundance lipids constitute key intermediates in lipid biosynthesis. In this study, we applied multiple lipidomic approaches, including high-sensitivity and high-specificity multiple reaction monitoring, to comprehensively quantify individual lipid species in crude palm oil. We also established a new liquid chromatography-coupled mass spectrometry method that allows direct quantification of low-abundance galactolipids in palm oil without the need for sample pretreatment. As crude palm oil contains large amounts of neutral lipids, our direct-detection method circumvents many of the challenges encountered with conventional lipid quantification methods. This approach allows direct measurement of lipids with no hassle during sample preparation and is more accurate and precise compared with other methods. PMID:24894356

  19. Interstrand cross-links are preferentially formed at the d(GC) sites in the reaction between cis-diamminedichloroplatinum (II) and DNA.

    PubMed

    Lemaire, M A; Schwartz, A; Rahmouni, A R; Leng, M

    1991-03-01

    A DNA restriction fragment with convergent SP6 and T7 promoters has undergone reaction with cis-diamminedichloroplatinum(II) (cis-DDP) and was then used as a template for RNA synthesis in vitro. The T7 and SP6 RNA polymerases generate fragments of defined sizes. Analysis of the RNA fragments shows that the polymerases are mainly blocked at the level of the d(GG) and d(AG) sites and to a lesser extent at the level of the d(GC) sites. The adducts at the d(GC) sites are more resistant to cyanide ion attack than those at the major sites and are identified as interstrand cross-links. The formation of an interstrand cross-link between the N-7 atoms of two guanine residues at the d(GC) sites was further confirmed by chemical modifications. PMID:2000402

  20. Identification of metE as a Second Target of the sRNA scr5239 in Streptomyces coelicolor

    PubMed Central

    Vockenhuber, Michael-Paul; Heueis, Nona; Suess, Beatrix

    2015-01-01

    While transcriptional regulation of the primary and secondary metabolism of the model organism Streptomyces coelicolor is well studied, little is still known about the role small noncoding RNAs (sRNAs) play in regulating gene expression in this organism. Here, we report the identification of a second target of the sRNA scr5239, an sRNA highly conserved in streptomycetes. The 159 nt long sRNA binds its target, the mRNA of the cobalamin independent methionine synthase metE (SCO0985), at the 5’ end of its open reading frame thereby repressing translation. We show that a high methionine level induces expression of scr5239 itself. This leads, in a negative feedback loop, to the repression of methionine biosynthesis. In contrast to the first reported target of this sRNA, the agarase dagA, this interaction seems to be conserved in a wide number of streptomycetes. PMID:25785836

  1. Experimental observation of silver and gold penetration into dental ceramic by means of a radiotracer technique

    SciTech Connect

    Moya, F.; Payan, J.; Bernardini, J.; Moya, E.G.

    1987-12-01

    A radiotracer technique was used to study silver and gold diffusion into dental porcelain under experimental conditions close to the real conditions in prosthetic laboratories for porcelain bakes. It was clearly shown that these non-oxidizable elements were able to diffuse into the ceramic as well as oxidizable ones. The penetration depth varied widely according to the element. The ratio DAg/DAu was about 10(3) around 850 degrees C. In contrast to gold, the silver diffusion rate was high enough to allow silver, from the metallic alloy, to be present at the external ceramic surface after diffusion into the ceramic. Hence, the greening of dental porcelains baked on silver-rich alloys could be explained mainly by a solid-state diffusion mechanism.

  2. Developments at the graft interface in homo- and hetero-grafts

    PubMed Central

    Clemente Moreno, Maria José; Hevin, Cyril; Ollat, Nathalie; Cookson, Sarah Jane

    2014-01-01

    Gene expression changes induced during graft union formation (the first month after grafting) in grapevine have been studied using whole genome microarrays. The genes differentially expressed between the rootstock and graft interface tissues of homo-grafts (Cabernet Sauvignon (CS) grafted onto CS) were compared at 3 and 28 days after grafting (dag). Graft union formation was associated with the upregulation of genes involved in secondary metabolism, cell wall, wound responses and hormone signaling. These gene expression differences were associated with the accumulation of lignin, cellulose and callose in the callus cells. Superimposed upon this, hetero-grafting between two different grapevine genotypes resulted in the further upregulation of stress and/or defense responses at the graft interface. Here we discuss the limitations of the techniques used to study the developments at the graft interface to date and future research directions to understand graft union formation in plants. PMID:24770337

  3. Comittees

    NASA Astrophysics Data System (ADS)

    2004-10-01

    Fritz Caspers (CERN, Switzerland), Michel Chanel (CERN, Switzerland), Håkan Danared (MSL, Sweden), Bernhard Franzke (GSI, Germany), Manfred Grieser (MPI für Kernphysik, Germany), Dieter Habs (LMU München, Germany), Jeffrey Hangst (University of Aarhus, Denmark), Takeshi Katayama (RIKEN/Univ. Tokyo, Japan), H.-Jürgen Kluge (GSI, Germany), Shyh-Yuan Lee (Indiana University, USA), Rudolf Maier (FZ Jülich, Germany), John Marriner (FNAL, USA), Igor Meshkov (JINR, Russia), Dieter Möhl (CERN, Switzerland), Vasily Parkhomchuk (BINP, Russia), Robert Pollock (Indiana University), Dieter Prasuhn (FZ Jülich, Germany), Dag Reistad (TSL, Sweden), John Schiffer (ANL, USA), Andrew Sessler (LBNL, USA), Alexander Skrinsky (BINP, Russia), Markus Steck (GSI, Germany), Jie Wei (BNL, USA), Andreas Wolf (MPI für Kernphysik, Germany), Hongwei Zhao (IMP, People's Rep. of China).

  4. Design of a Multi-mode Flight Deck Decision Support System for Airborne Conflict Management

    NASA Technical Reports Server (NTRS)

    Barhydt, Richard; Krishnamurthy, Karthik

    2004-01-01

    NASA Langley has developed a multi-mode decision support system for pilots operating in a Distributed Air-Ground Traffic Management (DAG-TM) environment. An Autonomous Operations Planner (AOP) assists pilots in performing separation assurance functions, including conflict detection, prevention, and resolution. Ongoing AOP design has been based on a comprehensive human factors analysis and evaluation results from previous human-in-the-loop experiments with airline pilot test subjects. AOP considers complex flight mode interactions and provides flight guidance to pilots consistent with the current aircraft control state. Pilots communicate goals to AOP by setting system preferences and actively probing potential trajectories for conflicts. To minimize training requirements and improve operational use, AOP design leverages existing alerting philosophies, displays, and crew interfaces common on commercial aircraft. Future work will consider trajectory prediction uncertainties, integration with the TCAS collision avoidance system, and will incorporate enhancements based on an upcoming air-ground coordination experiment.

  5. Protein kinase Calpha mediates a novel form of plasticity in the accessory olfactory bulb.

    PubMed

    Dong, C; Godwin, D W; Brennan, P A; Hegde, A N

    2009-10-20

    Modification of synapses in the accessory olfactory bulb (AOB) is believed to underlie pheromonal memory that enables mate recognition in mice. The memory, which is acquired with single-trial learning, forms only with coincident noradrenergic and glutamatergic inputs to the AOB. The mechanisms by which glutamate and norepinephrine (NE) alter the AOB synapses are not well understood. Here we present results that not only reconcile the earlier, seemingly contradictory, observations on the role of glutamate and NE in changing the AOB synapses, but also reveal novel mechanisms of plasticity. Our studies suggest that initially, glutamate acting at Group II metabotropic receptors and NE acting at alpha(2)-adrenergic receptors inhibit N-type and R-type Ca(2+) channels in mitral cells via a G-protein. The N-type and R-type Ca(2+) channel inhibition is reversed by activation of alpha(1)-adrenergic receptors and protein kinase Calpha (PKCalpha). Based on these results, we propose a hypothetical model for a new kind of synaptic plasticity in the AOB that accounts for the previous behavioral data on pheromonal memory. According to this model, initial inhibition of the Ca(2+) channels suppresses the GABAergic inhibitory feedback to mitral cells, causing disinhibition and Ca(2+) influx. NE also activates phospholipase C (PLC) through alpha(1)-adrenergic receptors generating inositol 1,4,5-trisphosphate and diacylglycerol (DAG). Calcium and DAG together activate PKCalpha which switches the disinhibition to increased inhibition of mitral cells. Thus, PKCalpha is likely to be a coincidence detector integrating glutamate and NE input in the AOB and bridging the short-term signaling to long-term structural changes resulting in enhanced inhibition of mitral cells that is thought to underlie memory formation. PMID:19580852

  6. A novel light-dependent activation of DAGK and PKC in bovine photoreceptor nuclei.

    PubMed

    Natalini, Paola M; Mateos, Melina V; Ilincheta de Boschero, Mónica G; Giusto, Norma M

    2014-08-01

    In this work, we describe a selective light-dependent distribution of the lipid kinase 1,2-diacylglycerol kinase (EC 2.7.1.107, DAGK) and the phosphorylated protein kinase C alpha (pPKCα) in a nuclear fraction of photoreceptor cells from bovine retinas. A nuclear fraction enriched in small nuclei from photoreceptor cells (PNF), was obtained when a modified nuclear isolation protocol developed by our laboratory was used. We measured and compared DAGK activity as phosphatidic acid (PA) formation in PNF obtained from retinas exposed to light and in retinas kept in darkness using [γ-(32)P]ATP or [(3)H]DAG. In the absence of exogenous substrates and detergents, no changes in DAGK activity were observed. However, when DAGK activity assays were performed in the presence of exogenous substrates, such as stearoyl arachidonoyl glycerol (SAG) or dioleoyl glycerol (DOG), and different detergents (used to make different DAGK isoforms evident), we observed significant light effects on DAGK activity, suggesting the presence of several DAGK isoforms in PNF. Under conditions favoring DAGKζ activity (DOG, Triton X-100, dioleoyl phosphatidylserine and R59022) we observed an increase in PA formation in PNF from retinas exposed to light with respect to those exposed to darkness. In contrast, under conditions favoring DAGKɛ (SAG, octylglucoside and R59022) we observed a decrease in its activity. These results suggest different physiological roles of the above-mentioned DAGK isoforms. Western blot analysis showed that whereas light stimulation of bovine retinas increases DAGKζ nuclear content, it decreases DAGKɛ and DAGKβ content in PNF. The role of PIP2-phospholipase C in light-stimulated DAGK activity was demonstrated using U73122. Light was also observed to induce enhanced pPKCα content in PNF. The selective distribution of DAGKζ and ɛ in PNF could be a light-dependent mechanism that in vertebrate retina promotes selective DAG removal and PKC regulation. PMID:24950064

  7. Activation of transient receptor potential canonical 3 (TRPC3)-mediated Ca2+ entry by A1 adenosine receptor in cardiomyocytes disturbs atrioventricular conduction.

    PubMed

    Sabourin, Jessica; Antigny, Fabrice; Robin, Elodie; Frieden, Maud; Raddatz, Eric

    2012-08-01

    Although the activation of the A(1)-subtype of the adenosine receptors (A(1)AR) is arrhythmogenic in the developing heart, little is known about the underlying downstream mechanisms. The aim of this study was to determine to what extent the transient receptor potential canonical (TRPC) channel 3, functioning as receptor-operated channel (ROC), contributes to the A(1)AR-induced conduction disturbances. Using embryonic atrial and ventricular myocytes obtained from 4-day-old chick embryos, we found that the specific activation of A(1)AR by CCPA induced sarcolemmal Ca(2+) entry. However, A(1)AR stimulation did not induce Ca(2+) release from the sarcoplasmic reticulum. Specific blockade of TRPC3 activity by Pyr3, by a dominant negative of TRPC3 construct, or inhibition of phospholipase Cs and PKCs strongly inhibited the A(1)AR-enhanced Ca(2+) entry. Ca(2+) entry through TRPC3 was activated by the 1,2-diacylglycerol (DAG) analog OAG via PKC-independent and -dependent mechanisms in atrial and ventricular myocytes, respectively. In parallel, inhibition of the atypical PKCζ by myristoylated PKCζ pseudosubstrate inhibitor significantly decreased the A(1)AR-enhanced Ca(2+) entry in both types of myocytes. Additionally, electrocardiography showed that inhibition of TRPC3 channel suppressed transient A(1)AR-induced conduction disturbances in the embryonic heart. Our data showing that A(1)AR activation subtly mediates a proarrhythmic Ca(2+) entry through TRPC3-encoded ROC by stimulating the phospholipase C/DAG/PKC cascade provide evidence for a novel pathway whereby Ca(2+) entry and cardiac function are altered. Thus, the A(1)AR-TRPC3 axis may represent a potential therapeutic target. PMID:22692208

  8. Solution structures of purine base analogues 9-deazaguanine and 9-deazahypoxanthine.

    PubMed

    Karnawat, Vishakha; Puranik, Mrinalini

    2016-03-01

    Deaza analogues of nucleobases are potential drugs against infectious diseases caused by parasites. A caveat is that apart from binding their target parasite enzymes, they also bind and inhibit enzymes of the host. In order to design derivatives of deaza analogues which specifically bind target enzymes, knowledge of their molecular structure, protonation state, and predominant tautomers at physiological conditions is essential. We have employed resonance Raman spectroscopy at an excitation wavelength of 260 nm, to decipher solution structure of 9-deazaguanine (9DAG) and 9-deazahypoxanthine (9DAH). These are analogues of guanine and hypoxanthine, respectively, and have been exploited to study static complexes of nucleobase binding enzymes. Such enzymes are known to perturb pKa of their ligands, and thus, we also determined solution structures of these analogues at two, acidic and alkaline, pH. Structure of each possible protonation state and tautomer was computed using density functional theoretical calculations. Species at various pHs were identified based on isotopic shifts in experimental wavenumbers and by comparing these shifts with corresponding computed isotopic shifts. Our results show that at physiological pH, N1 of pyrimidine ring in 9DAG and 9DAH bears a proton. At lower pH, N3 is place of protonation, and at higher pH, deprotonation occurs at N1 position. The proton at N7 of purine ring remains intact even at pH 12.5. We have further compared these results with naturally occurring nucleotides. Our results identify key vibrational modes which can report on hydrogen bonding interactions, protonation and deprotonation in purine rings upon binding to the active site of enzymes. PMID:25894214

  9. The role of diacylglycerol kinase ζ and phosphatidic acid in the mechanical activation of mammalian target of rapamycin (mTOR) signaling and skeletal muscle hypertrophy.

    PubMed

    You, Jae-Sung; Lincoln, Hannah C; Kim, Chan-Ran; Frey, John W; Goodman, Craig A; Zhong, Xiao-Ping; Hornberger, Troy A

    2014-01-17

    The activation of mTOR signaling is essential for mechanically induced changes in skeletal muscle mass, and previous studies have suggested that mechanical stimuli activate mTOR (mammalian target of rapamycin) signaling through a phospholipase D (PLD)-dependent increase in the concentration of phosphatidic acid (PA). Consistent with this conclusion, we obtained evidence which further suggests that mechanical stimuli utilize PA as a direct upstream activator of mTOR signaling. Unexpectedly though, we found that the activation of PLD is not necessary for the mechanically induced increases in PA or mTOR signaling. Motivated by this observation, we performed experiments that were aimed at identifying the enzyme(s) that promotes the increase in PA. These experiments revealed that mechanical stimulation increases the concentration of diacylglycerol (DAG) and the activity of DAG kinases (DGKs) in membranous structures. Furthermore, using knock-out mice, we determined that the ζ isoform of DGK (DGKζ) is necessary for the mechanically induced increase in PA. We also determined that DGKζ significantly contributes to the mechanical activation of mTOR signaling, and this is likely driven by an enhanced binding of PA to mTOR. Last, we found that the overexpression of DGKζ is sufficient to induce muscle fiber hypertrophy through an mTOR-dependent mechanism, and this event requires DGKζ kinase activity (i.e. the synthesis of PA). Combined, these results indicate that DGKζ, but not PLD, plays an important role in mechanically induced increases in PA and mTOR signaling. Furthermore, this study suggests that DGKζ could be a fundamental component of the mechanism(s) through which mechanical stimuli regulate skeletal muscle mass. PMID:24302719

  10. Phospholipase C Epsilon (PLCε) Induced TRPC6 Activation: A Common but Redundant Mechanism in Primary Podocytes

    PubMed Central

    Kalwa, Hermann; Storch, Ursula; Demleitner, Jana; Fiedler, Susanne; Mayer, Tim; Kannler, Martina; Fahlbusch, Meike; Barth, Holger; Smrcka, Alan; Hildebrandt, Friedhelm; Gudermann, Thomas; Dietrich, Alexander

    2016-01-01

    In eukaryotic cells, activation of phospholipase C (PLC)-coupled membrane receptors by hormones leads to an increase in the intracellular Ca2+ concentration [Ca2+]i. Catalytic activity of PLCs results in the hydrolysis of phosphatidylinositol 4,5-bisphosphate to generate inositol 1,4,5-trisphosphate (IP3) and diacylglycerol (DAG) which opens DAG-sensitive classical transient receptor channels 3, 6, and 7 (TRPC3/6/7), initiating Ca2+ influx from the extracellular space. Patients with focal segmental glomerulosclerosis (FSGS) express gain-of-function mutants of TRPC6, while others carry loss-of-function mutants of PLCε, raising the intriguing possibility that both proteins interact and might work in the same signalling pathway. While TRPC6 activation by PLCβ and PLCγ isozymes was extensively studied, the role of PLCε in TRPC6 activation remains elusive. TRPC6 was co-immunoprecipitated with PLCε in a heterologous overexpression system in HEK293 cells as well as in freshly isolated murine podocytes. Receptor-operated TRPC6 currents in HEK293 cells expressing TRPC6 were reduced by a specific PLCε siRNA and by a PLCε loss-of-function mutant isolated from a patient with FSGS. PLCε-induced TRPC6 activation was also identified in murine embryonic fibroblasts (MEFs) lacking Gαq/11 proteins. Further analysis of the signal transduction pathway revealed a Gα12/13 Rho-GEF activation which induced Rho-mediated PLCε stimulation. Therefore, we identified a new pathway for TRPC6 activation by PLCε. PLCε-/- podocytes however, were undistinguishable from WT podocytes in their angiotensin II-induced formation of actin stress fibers and their GTPγS-induced TRPC6 activation, pointing to a redundant role of PLCε-mediated TRPC6 activation at least in podocytes. PMID:25521631

  11. Can We Stop Antibiotic Therapy When Signs and Symptoms Have Resolved in Diabetic Foot Infection Patients?

    PubMed Central

    Chu, Yuejie; Wang, Chao; Zhang, Jinghang; Wang, Penghua; Xu, Jun; Ding, Min; Li, Xiwen; Hou, Xiaoli; Feng, Shuhong; Li, Xuemei

    2015-01-01

    The study aimed to investigate whether we can stop antibiotic therapy when signs and symptoms have resolved in diabetic foot infection (DFI) patients with different grades of peripheral arterial disease (PAD) and those without PAD, and to determine whether the severity of PAD and infection has an effect on antibiotic therapy duration. A prospective randomized controlled trial of DFI patients was carried out. Patients were randomized into 2 groups when signs and symptoms had resolved: continuing antibiotics group (CAG) and discontinuing antibiotics group (DAG). The recurrence and clinical outcomes were recorded. The recurrence rate of mild infection with mild/moderate PAD was similar in the 2 groups. Compared with CAG, the recurrence rate of mild infection with severe PAD was higher in DAG (P = .030), also for moderate/severe infection with PAD (mild/moderate [P = .032]; severe [P = .008]). No difference was found in the 2 groups (either mild or moderate/severe) for those without PAD. The clinical outcomes of mild infection in patients were similar in the 2 groups. For moderate/severe infection, the healing rate was higher (73.3% vs 48.3%), and the rate of minor/major amputation and death was lower (23.8% vs 49.4%; 6.9% vs 20.7%; 2.0% vs 13.8%) in the CAG. When the clinical signs and symptoms of infection have resolved, it might be appropriate to stop antibiotics for DFI patients without PAD, and also for patients with mild infection with mild/moderate PAD. For patients with mild infection with severe PAD and moderate/severe infection with PAD, we should perhaps continue antibiotic treatment. Continuing antibiotic therapy could improve clinical outcomes for patients with moderate/severe infection. PMID:26248828

  12. Emplacement of volcanic vents and geodynamics of Central Anatolia, Turkey

    NASA Astrophysics Data System (ADS)

    Dhont, D.; Chorowicz, J.; Yürür, T.; Froger, J.-L.; Köse, O.; Gündogdu, N.

    1998-10-01

    Observations on Synthetic Aperture Radar (SAR) scenes of the European Remote Sensing (ERS) satellite and Digital Elevation Models (DEMs), complemented by field structural analysis permit a new understanding of relationships between tectonics and volcanism since the late Miocene (10 Ma) in Central Anatolia. Volcanic edifices form elongate stratovolcanoes, linear clusters and volcanic ridges. They indicate emplacement on tension fractures and tail-crack or horsetail features. For instance, the Kara Dag volcano is rooted on a tail-crack which accommodates a horizontal left-lateral throw component at a fault termination. Caldera complexes of Cappadocia are associated with horsetail fault patterns. The emplacement of volcanoes also benefits from larger-scale tectonic structures: the Erciyes Dag volcano is localized by the Sultan Saz releasing bend which opens along the sinistral strike-slip Ecemis fault. Deformation has been analysed from tension fractures—which are perpendicular to the direction of extension—and from field structural analysis. On a regional scale, the tectonic regime responsible for the distribution of volcanic vents in this area of convergence and lateral extrusion, is not compression but extension. The Central Taurus range is the thermally uplifted shoulder of the Adana-Cilicia basin, which is related to lithosphere thinning. Westward movements in the northwestern part of the studied area are influenced by the active back-arc Aegean extension situated to the west. Farther to the south, the direction of motion turns southwest and south, under the influence of the opening of the Adana-Cilicia basin. We interpreted that extension in the Central Anatolian plateau is related to crustal blocks moving above sub-horizontal detachment surfaces located in the lower crust. This is based on several facts: the Tuz Gölü fault zone is a within-crust detachment; the Tuz Gölü basin does not affect the whole lithosphere because otherwise it would have been

  13. Planetary Bioresources and Astroecology. 1. Planetary Microcosm Bioassays of Martian and Carbonaceous Chondrite Materials: Nutrients, Electrolyte Solutions, and Algal and Plant Responses

    NASA Astrophysics Data System (ADS)

    Mautner, Michael N.

    2002-07-01

    The biological fertilities of planetary materials can be assessed using microcosms based on meteorites. This study applies microcosm tests to martian meteorites and analogues and to carbonaceous chondrites. The biological fertilities of these materials are rated based on the soluble electrolyte nutrients, the growth of mesophile and cold-tolerant algae, and plant tissue cultures. The results show that the meteorites, in particular the Murchison CM2 carbonaceous chondrite and DaG 476 martian shergottite, contain high levels of water-extractable Ca, Mg, and SO 4-S. The martian meteorites DaG 476 and EETA 79001 also contain higher levels of extractable essential nutrients NO 3-N (0.013-0.017 g kg -1) and PO 4-P (0.019-0.046 g kg -1) than the terrestrial analogues. The yields of most of the water-extractable electrolytes vary only by factors of 2-3 under a wide range of planetary conditions. However, the long-term extractable phosphate increases significantly under a CO 2 atmosphere. The biological yields of algae and plant tissue cultures correlate with extractable NO 3-N and PO 4-P, identifying these as the limiting nutrients. Mesophilic algae and Asparagus officinalis cultures are identified as useful bioassay agents. A fertility rating system based on microcosm tests is proposed. The results rate the fertilities in the order martian basalts > terrestrial basalt, agricultural soil > carbonaceous chondrites, lava ash > cumulate igneous rock. The results demonstrate the application of planetary microcosms in experimental astroecology to rate planetary materials as targets for astrobiology exploration and as potential space bioresources. For example, the extractable materials in Murchison suggest that concentrated internal solutions in carbonaceous asteroids (3.8 mol L -1 electrolytes and 10 g L -1 organics) can support and disperse microorganisms introduced by natural or directed panspermia in early solar systems. The results also suggest that carbonaceous asteroids

  14. Positive Effects of Voluntary Running on Metabolic Syndrome-Related Disorders in Non-Obese Hereditary Hypertriacylglycerolemic Rats

    PubMed Central

    Škop, Vojtěch; Malínská, Hana; Trnovská, Jaroslava; Hüttl, Martina; Cahová, Monika; Blachnio-Zabielska, Agnieszka; Baranowski, Marcin; Burian, Martin; Oliyarnyk, Olena; Kazdová, Ludmila

    2015-01-01

    While metabolic syndrome is often associated with obesity, 25% of humans suffering from it are not obese and the effect of physical activity remains unclear in such cases. Therefore, we used hereditary hypertriaclyglycerolemic (HHTg) rats as a unique model for studying the effect of spontaneous physical activity [voluntary running (VR)] on metabolic syndrome-related disorders, such as dyslipidemia, in non-obese subjects. Adult HHTg males were fed standard (CD) or high-sucrose (HSD) diets ad libitum for four weeks. Within both dietary groups, some of the rats had free access to a running wheel (CD+VR, HSD+VR), whereas the controls (CD, HSD) had no possibility of extra physical activity. At the end of the four weeks, we measured the effects of VR on various metabolic syndrome-associated parameters: (i) biochemical parameters, (ii) the content and composition of triacylglycerols (TAG), diacylglycerols (DAG), ceramides and membrane phospholipids, and (iii) substrate utilization in brown adipose tissue. In both dietary groups, VR led to various positive effects: reduced epididymal and perirenal fat depots; increased epididymal adipose tissue lipolysis; decreased amounts of serum TAG, non-esterified fatty acids and insulin; a higher insulin sensitivity index. While tissue ceramide content was not affected, decreased TAG accumulation resulted in reduced and modified liver, heart and skeletal muscle DAG. VR also had a beneficial effect on muscle membrane phospholipid composition. In addition, compared with the CD group, the CD+VR rats exhibited increased fatty acid oxidation and protein content in brown adipose tissue. Our results confirm that physical activity in a non-obese model of severe dyslipidemia has many beneficial effects and can even counteract the negative effects of sucrose consumption. Furthermore, they suggest that the mechanism by which these effects are modulated involves a combination of several positive changes in lipid metabolism. PMID:25830228

  15. How did nature engineer the highest surface lipid accumulation among plants? Exceptional expression of acyl-lipid-associated genes for the assembly of extracellular triacylglycerol by Bayberry (Myrica pensylvanica) fruits.

    PubMed

    Simpson, Jeffrey P; Thrower, Nicholas; Ohlrogge, John B

    2016-09-01

    Bayberry (Myrica pensylvanica) fruits are covered with a remarkably thick layer of crystalline wax consisting of triacylglycerol (TAG) and diacylglycerol (DAG) esterified exclusively with saturated fatty acids. As the only plant known to accumulate soluble glycerolipids as a major component of surface waxes, Bayberry represents a novel system to investigate neutral lipid biosynthesis and lipid secretion by vegetative plant cells. The assembly of Bayberry wax is distinct from conventional TAG and other surface waxes, and instead proceeds through a pathway related to cutin synthesis (Simpson and Ohlrogge, 2016). In this study, microscopic examination revealed that the fruit tissue that produces and secretes wax (Bayberry knobs) is fully developed before wax accumulates and that wax is secreted to the surface without cell disruption. Comparison of transcript expression to genetically related tissues (Bayberry leaves, M. rubra fruits), cutin-rich tomato and cherry fruit epidermis, and to oil-rich mesocarp and seeds, revealed exceptionally high expression of 13 transcripts for acyl-lipid metabolism together with down-regulation of fatty acid oxidases and desaturases. The predicted protein sequences of the most highly expressed lipid-related enzyme-encoding transcripts in Bayberry knobs are 100% identical to the sequences from Bayberry leaves, which do not produce surface DAG or TAG. Together, these results indicate that TAG biosynthesis and secretion in Bayberry is achieved by both up and down-regulation of a small subset of genes related to the biosynthesis of cutin and saturated fatty acids, and also implies that modifications in gene expression, rather than evolution of new gene functions, was the major mechanism by which Bayberry evolved its specialized lipid metabolism. This article is part of a Special Issue entitled: Plant Lipid Biology edited by Kent D. Chapman and Ivo Feussner. PMID:26869450

  16. Comparison of postprandial lipaemia between native and palm random esterified acid oils in two different monogastric species (dogs and broiler chickens).

    PubMed

    Fragua, V; Vilarrasa, E; Manzanilla, E G; Villaverde, C; Barroeta, A C

    2013-05-01

    It has been reported that applying a chemical reesterification process to a native fat results in a new fat source with different physicochemical properties due to their different fatty acid (FA) positional distribution within the glycerol moiety and their different proportions of mono (MAG)-, di (DAG)- and triacylglycerides (TAG). Thus, this reesterification could affect fat digestion, absorption and metabolism; and this effect could vary among species given their differences in fat metabolism. The aim of this study was to compare the effect of feeding two different random esterified acid oils (REAO), presenting different MAG, DAG and TAG proportions, with their corresponding native oil on postprandial lipaemia in broiler chickens and dogs. For this purpose, 18 dogs and 54 broiler chickens were fed a basal diet supplemented with palm native oil, palm REAO low MAG or palm REAO high MAG. The inclusion level of the oils was 10% of the diet in dogs and 6% in broiler chickens. Serum postprandial TAG concentration (mg/dl) after feeding a single meal was measured at different time points during 12 h in dogs and 3 h in chickens. Although fasting serum TAG concentration values were similar for both species (47 ± 2.4 mg/dl for dogs and 44 ± 3.0 mg/dl for broilers; p = 0.522), postprandial TAG concentrations tended to be higher in broilers than in dogs (p = 0.058). Treatment had no effect on TAG concentration at any time point in any species (p = 0.768 for dogs, p = 0.947 for broilers). However, the postprandial TAG curves were very different between species; in broiler chickens, TAG concentration returned to the fasting values 3 h after feeding while in dogs, the TAG concentration still had not returned to basal levels 12 h after feeding. PMID:23639020

  17. Enterococcus faecalis Glycolipids Modulate Lipoprotein-Content of the Bacterial Cell Membrane and Host Immune Response

    PubMed Central

    Otto, Andreas; Sava, Irina G.; Wobser, Dominique; Bao, Yinyin; Hese, Katrin; Broszat, Melanie; Henneke, Philipp; Becher, Dörte; Huebner, Johannes

    2015-01-01

    In this study, we investigated the impact of the cell membrane composition of E. faecalis on its recognition by the host immune system. To this end, we employed an E. faecalis deletion mutant (ΔbgsA) that does not synthesize the major cell membrane glycolipid diglycosyl-diacylglycerol (DGlcDAG). Proteomic analysis revealed that 13 of a total of 21 upregulated surface-associated proteins of E. faecalis ΔbgsA were lipoproteins. This led to a total lipoprotein content in the cell membrane of 35.8% in ΔbgsA compared to only 9.4% in wild-type bacteria. Increased lipoprotein content strongly affected the recognition of ΔbgsA by mouse macrophages in vitro with an increased stimulation of TNF-α production by heat-fixed bacteria and secreted antigens. Inactivation of the prolipoprotein diacylglycerol transferase (lgt) in ΔbgsA abrogated TNF-α induction by a ΔbgsA_lgt double mutant indicating that lipoproteins mediate increased activation of mouse macrophages by ΔbgsA. Heat-fixed ΔbgsA bacteria, culture supernatant, or cell membrane lipid extract activated transfected HEK cells in a TLR2-dependent fashion; the same was not true of wild-type bacteria. In mice infected intraperitoneally with a sublethal dose of E. faecalis we observed a 70% greater mortality in mice infected with ΔbgsA compared with wild-type-infected mice. Increased mortality due to ΔbgsA infection was associated with elevated plasma levels of the inflammatory cytokines TNF-α, IL-6 and MIP-2. In summary, our results provide evidence that an E. faecalis mutant lacking its major bilayer forming glycolipid DGlcDAG upregulates lipoprotein expression leading to increased activation of the host innate immune system and virulence in vivo. PMID:26172831

  18. Carnitine palmitoyltransferase-1 up-regulation by PPAR-β/δ prevents lipid-induced endothelial dysfunction.

    PubMed

    Toral, Marta; Romero, Miguel; Jiménez, Rosario; Mahmoud, Ayman Moawad; Barroso, Emma; Gómez-Guzmán, Manuel; Sánchez, Manuel; Cogolludo, Ángel; García-Redondo, Ana B; Briones, Ana M; Vázquez-Carrera, Manuel; Pérez-Vizcaíno, Francisco; Duarte, Juan

    2015-11-01

    Fatty acids cause endothelial dysfunction involving increased ROS (reactive oxygen species) and reduced NO (nitric oxide) bioavailability. We show that in MAECs (mouse aortic endothelial cells), the PPARβ/δ (peroxisome- proliferator-activated receptor β/δ) agonist GW0742 prevented the decreased A23187-stimulated NO production, phosphorylation of eNOS (endothelial nitric oxide synthase) at Ser1177 and increased intracellular ROS levels caused by exposure to palmitate in vitro. The impaired endothelium-dependent relaxation to acetylcholine in mouse aorta induced by palmitate was restored by GW0742. In vivo, GW0742 treatment prevented the reduced aortic relaxation, phosphorylation of eNOS at Ser1177, and increased ROS production and NADPH oxidase in mice fed on a high-fat diet. The PPARβ/δ antagonist GSK0660 abolished all of these protective effects induced by GW0742. This agonist enhanced the expression of CPT (carnitine palmitoyltransferase)-1. The effects of GW0742 on acetylcholine- induced relaxation in aorta and on NO and ROS production in MAECs exposed to palmitate were abolished by the CPT-1 inhibitor etomoxir or by siRNA targeting CPT-1. GW0742 also inhibited the increase in DAG (diacylglycerol), PKCα/βII (protein kinase Cα/βII) activation, and phosphorylation of eNOS at Thr495 induced by palmitate in MAECs, which were abolished by etomoxir. In conclusion, PPARβ/δ activation restored the lipid-induced endothelial dysfunction by up-regulation of CPT-1, thus reducing DAG accumulation and the subsequent PKC-mediated ROS production and eNOS inhibition. PMID:26253087

  19. Characterization of Human Duodenal Fluids in Fasted and Fed State Conditions.

    PubMed

    Riethorst, Danny; Mols, Raf; Duchateau, Guus; Tack, Jan; Brouwers, Joachim; Augustijns, Patrick

    2016-02-01

    This work provides an elaborate characterization of human intestinal fluids (HIF) collected in fasted- and fed-state conditions. HIF from 20 healthy volunteers (10 M/F) were aspirated by intubation near the ligament of Treitz in a time-dependent manner (10-min intervals) and characterized for pH, bile salts, phospholipids, cholesterol, triacylglycerides (TAG), diacylglycerides (DAG), monoacylglycerides (MAG), free fatty acids (FFA), pancreatic lipase, phospholipase A2, and nonspecific esterase activity. For almost all parameters, a food-induced effect was observed. Results were characterized by a high variability, as illustrated by the broad ranges observed for each parameter: pH (fasted: 3.4-8.3; fed: 4.7-7.1), bile salts (fasted: 0.03-36.18 mM; fed: 0.74-86.14 mM), phospholipids (fasted: 0.01-6.33 mM; fed: 0.16-14.39 mM), cholesterol (fasted: 0-0.48 mM; fed: 0-3.29 mM), TAG (fed: 0-6.76 mg/mL), DAG (fed: 0-3.64 mg/mL), MAG (fasted: 0-1.09 mg/mL; fed: 0-11.36 mg/mL), FFA (fasted: 0-3.86 mg/mL; fed: 0.53-15.0 mg/mL), pancreatic lipase (fasted: 26-86 g/mL; fed: 146-415 g/mL), phospholipase A2 (fasted: 3-6 ng/mL; fed: 4.3-27.7 ng/mL), and nonspecific esterase activity (fasted: 270-4900 U/mL; fed: 430-4655 U/mL). This comprehensive overview may serve as reference data for physiologically based pharmacokinetic modeling and the optimization of biorelevant simulated intestinal fluids for the use in in vitro dissolution, solubility, and permeability profiling. PMID:26228456

  20. Transcriptional and biochemical responses of monoacylglycerol acyltransferase-mediated oil synthesis and associated senescence-like responses in Nicotiana benthamiana

    PubMed Central

    Divi, Uday K.; El Tahchy, Anna; Vanhercke, Thomas; Petrie, James R.; Robles-Martinez, Jose A.; Singh, Surinder P.

    2014-01-01

    Triacylglycerol (TAG) accumulates in plant seeds as a major renewable source of carbon for food, fuel and industrial feedstock. Approaches to enhance TAG content by altering lipid pathways and genes in vegetative parts have gained significant attention for biofuel and other applications. However, consequences of these modifications are not always studied in detail. In an attempt to increase TAG levels in leaves we previously demonstrated that a novel substrate, monoacylglycerol (MAG), can be used for the biosynthesis of diacylglycerol (DAG) and TAG. Transient expression of the Mus musculus monoacylglycerol acyltransferases MGAT1 and 2 in the model plant Nicotiana benthamiana increased TAG levels at 5 days post-infiltration (dpi). Here we show that increased TAG and DAG levels can be achieved as early as 2 dpi. In addition, the MGAT1 infiltrated areas showed senescence-like symptoms from 3 dpi onwards. To unravel underlying molecular mechanisms, Illumina deep sequencing was carried out (a) for de-novo assembling and annotation of N. benthamiana leaf transcripts and (b) to characterize MGAT1-responsive transcriptome. We found that MGAT1-responsive genes are involved in several processes including TAG biosynthesis, photosynthesis, cell-wall, cutin, suberin, wax and mucilage biosynthesis, lipid and hormone metabolism. Comparative analysis with transcript profiles from other senescence studies identified characteristic gene expression changes involved in senescence induction. We confirmed that increased TAG and observed senescence-symptoms are due to the MAG depletion caused by MGAT1 activity and suggest a mechanism for MGAT1 induced TAG increase and senescence-like symptoms. The data generated will serve as a valuable resource for oil and senescence related studies and for future N. benthamiana transcriptome studies. PMID:24904604

  1. Transcriptional and biochemical responses of monoacylglycerol acyltransferase-mediated oil synthesis and associated senescence-like responses in Nicotiana benthamiana.

    PubMed

    Divi, Uday K; El Tahchy, Anna; Vanhercke, Thomas; Petrie, James R; Robles-Martinez, Jose A; Singh, Surinder P

    2014-01-01

    Triacylglycerol (TAG) accumulates in plant seeds as a major renewable source of carbon for food, fuel and industrial feedstock. Approaches to enhance TAG content by altering lipid pathways and genes in vegetative parts have gained significant attention for biofuel and other applications. However, consequences of these modifications are not always studied in detail. In an attempt to increase TAG levels in leaves we previously demonstrated that a novel substrate, monoacylglycerol (MAG), can be used for the biosynthesis of diacylglycerol (DAG) and TAG. Transient expression of the Mus musculus monoacylglycerol acyltransferases MGAT1 and 2 in the model plant Nicotiana benthamiana increased TAG levels at 5 days post-infiltration (dpi). Here we show that increased TAG and DAG levels can be achieved as early as 2 dpi. In addition, the MGAT1 infiltrated areas showed senescence-like symptoms from 3 dpi onwards. To unravel underlying molecular mechanisms, Illumina deep sequencing was carried out (a) for de-novo assembling and annotation of N. benthamiana leaf transcripts and (b) to characterize MGAT1-responsive transcriptome. We found that MGAT1-responsive genes are involved in several processes including TAG biosynthesis, photosynthesis, cell-wall, cutin, suberin, wax and mucilage biosynthesis, lipid and hormone metabolism. Comparative analysis with transcript profiles from other senescence studies identified characteristic gene expression changes involved in senescence induction. We confirmed that increased TAG and observed senescence-symptoms are due to the MAG depletion caused by MGAT1 activity and suggest a mechanism for MGAT1 induced TAG increase and senescence-like symptoms. The data generated will serve as a valuable resource for oil and senescence related studies and for future N. benthamiana transcriptome studies. PMID:24904604

  2. G protein-coupled receptor (GPR)40-dependent potentiation of insulin secretion in mouse islets is mediated by protein kinase D1

    PubMed Central

    Ferdaoussi, M.; Bergeron, V.; Zarrouki, B.; Kolic, J.; Cantley, J.; Fielitz, J.; Olson, E. N.; Prentki, M.; Biden, T.; MacDonald, P. E.

    2013-01-01

    Aims/hypothesis Activation of the G protein-coupled receptor (GPR)40 by long-chain fatty acids potentiates glucose-stimulated insulin secretion (GSIS) from pancreatic beta cells, and GPR40 agonists are in clinical development for type 2 diabetes therapy. GPR40 couples to the G protein subunit Gαq/11 but the signalling cascade activated down-stream is unknown. This study aimed to determine the mechanisms of GPR40-dependent potentiation of GSIS by fatty acids. Methods Insulin secretion in response to glucose, oleate or diacylglycerol (DAG) was assessed in dynamic perifusions and static incubations in islets from wild-type (WT) and Gpr40−/− mice. Depolymerisation of filamentous actin (F-actin) was visualised by phalloidin staining and epifluorescence. Pharmacological and molecular approaches were used to ascertain the roles of protein kinase D (PKD) and protein kinase C delta in GPR40-mediated potentiation of GSIS. Results Oleate potentiates the second phase of GSIS, and this effect is largely dependent upon GPR40. Accordingly, oleate induces rapid F-actin remodelling in WT but not in Gpr40−/− islets. Exogenous DAG potentiates GSIS in both WT and Gpr40−/− islets. Oleate induces PKD phosphorylation at residues Ser-744/748 and Ser-916 in WT but not Gpr40−/− islets. Importantly, oleate-induced F-actin depolymerisation and potentiation of GSIS are lost upon pharmacological inhibition of PKD1 or deletion of Prkd1. Conclusions/interpretation We conclude that the signalling cascade downstream of GPR40 activation by fatty acids involves activation of PKD1, F-actin depolymerisation and potentiation of second-phase insulin secretion. These results provide important information on the mechanisms of action of GPR40, a novel drug target for type 2 diabetes. PMID:22820510

  3. Palmitate induces ER calcium depletion and apoptosis in mouse podocytes subsequent to mitochondrial oxidative stress.

    PubMed

    Xu, S; Nam, S M; Kim, J-H; Das, R; Choi, S-K; Nguyen, T T; Quan, X; Choi, S J; Chung, C H; Lee, E Y; Lee, I-K; Wiederkehr, A; Wollheim, C B; Cha, S-K; Park, K-S

    2015-01-01

    Pathologic alterations in podocytes lead to failure of an essential component of the glomerular filtration barrier and proteinuria in chronic kidney diseases. Elevated levels of saturated free fatty acid (FFA) are harmful to various tissues, implemented in the progression of diabetes and its complications such as proteinuria in diabetic nephropathy. Here, we investigated the molecular mechanism of palmitate cytotoxicity in cultured mouse podocytes. Incubation with palmitate dose-dependently increased cytosolic and mitochondrial reactive oxygen species, depolarized the mitochondrial membrane potential, impaired ATP synthesis and elicited apoptotic cell death. Palmitate not only evoked mitochondrial fragmentation but also caused marked dilation of the endoplasmic reticulum (ER). Consistently, palmitate upregulated ER stress proteins, oligomerized stromal interaction molecule 1 (STIM1) in the subplasmalemmal ER membrane, abolished the cyclopiazonic acid-induced cytosolic Ca(2+) increase due to depletion of luminal ER Ca(2+). Palmitate-induced ER Ca(2+) depletion and cytotoxicity were blocked by a selective inhibitor of the fatty-acid transporter FAT/CD36. Loss of the ER Ca(2+) pool induced by palmitate was reverted by the phospholipase C (PLC) inhibitor edelfosine. Palmitate-dependent activation of PLC was further demonstrated by following cytosolic translocation of the pleckstrin homology domain of PLC in palmitate-treated podocytes. An inhibitor of diacylglycerol (DAG) kinase, which elevates cytosolic DAG, strongly promoted ER Ca(2+) depletion by low-dose palmitate. GF109203X, a PKC inhibitor, partially prevented palmitate-induced ER Ca(2+) loss. Remarkably, the mitochondrial antioxidant mitoTEMPO inhibited palmitate-induced PLC activation, ER Ca(2+) depletion and cytotoxicity. Palmitate elicited cytoskeletal changes in podocytes and increased albumin permeability, which was also blocked by mitoTEMPO. These data suggest that oxidative stress caused by saturated FFA

  4. New noble gas data of primitive and differentiated achondrites including Northwest Africa 011 and Tafassasset

    NASA Astrophysics Data System (ADS)

    Patzer, A.; Schultz, L.; Franke, L.

    2003-10-01

    This work reports on the noble gas inventory of 3 new acapulcoites, 3 brachinites, 2 new eucrites from the Dar al Gani region in Libya, the unique achondrite Dar al Gani (DaG) 896 from the same locality, the new eucrite-like achondrite Northwest Africa (NWA) 011, and the controversial sample Tafassasset. We determined cosmic ray exposure and gas retention ages, evaluated shielding conditions, and discuss the trapped noble gas component of the specimens. All exposure ages are within the known range of stony meteorites and partly confirm previously established age clusters. Shielding conditions vary, suggesting substantial shielding for all 3 brachinites and Tafassasset. We cannot exclude, however, that the Mg-rich composition of brachinites simply simulates heavy shielding. Regarding the trapped component, we found Q-like compositions only for the acapulcoite Thiel Mountains (TIL) 99002. The brachinite Elephant Moraine (EET) 99402 yields a high, subsolar 36Ar/ 132Xe ratio of ~400 along with a slightly elevated 84Kr/132Xe ratio, indicating minor atmospheric contamination. All the other samples, particularly the eucrite DaG 983, are characterized by clearly elevated Ar/Kr/Xe ratios due to significant terrestrial alteration. Tafassasset exhibits noble gas parameters that are different from those of CR chondrites, including a relatively high cosmic ray exposure age, the absence of a solar component, low 132Xe concentrations, a low trapped 36Ar/132Xe ratio of ~30, and a noticeable amount of radiogenic 129Xe. Similar attributes have been observed for some primitive achondrites. These attributes are also consistent with the metamorphic character of the sample. We, therefore, consider Tafassasset's noble gas record to be inconclusive as to its classification (primitive achondrite versus metamorphosed CR chondrite).

  5. Light-Driven Overall Water Splitting Enabled by a Photo-Dember Effect Realized on 3D Plasmonic Structures.

    PubMed

    Chen, Min; Gu, Jiajun; Sun, Cheng; Zhao, Yixin; Zhang, Ruoxi; You, Xinyuan; Liu, Qinglei; Zhang, Wang; Su, Yishi; Su, Huilan; Zhang, Di

    2016-07-26

    Photoelectric conversion driven by sunlight has a broad range of energy/environmental applications (e.g., in solar cells and water splitting). However, difficulties are encountered in the separation of photoexcited charges. Here, we realize a long-range (∼1.5 μm period) electric polarization via asymmetric localization of surface plasmons on a three-dimensional silver structure (3D-Ag). This visible-light-responsive effect-the photo-Dember effect, can be analogous to the thermoelectric effect, in which hot carriers are thermally generated instead of being photogenerated. The induced electric field can efficiently separate photogenerated charges, enabling sunlight-driven overall water splitting on a series of dopant-free commercial semiconductor particles (i.e., ZnO, CeO2, TiO2, and WO3) once they are combined with the 3D-Ag substrate. These photocatalytic processes can last over 30 h on 3D-Ag+ZnO, 3D-Ag+CeO2, and 3D-Ag+TiO2, thus demonstrating good catalytic stability for these systems. Using commercial WO3 powder as a reference, the amount of O2 generated with 3D-Ag+CeO2 surpasses even its recently reported counterpart in which sacrificial reagents had to be involved to run half-reactions. This plasmon-mediated charge separation strategy provides an effective way to improve the efficiency of photoelectric energy conversion, which can be useful in photovoltaics and photocatalysis. PMID:27351779

  6. Bayesian Estimation and Inference Using Stochastic Electronics

    PubMed Central

    Thakur, Chetan Singh; Afshar, Saeed; Wang, Runchun M.; Hamilton, Tara J.; Tapson, Jonathan; van Schaik, André

    2016-01-01

    In this paper, we present the implementation of two types of Bayesian inference problems to demonstrate the potential of building probabilistic algorithms in hardware using single set of building blocks with the ability to perform these computations in real time. The first implementation, referred to as the BEAST (Bayesian Estimation and Stochastic Tracker), demonstrates a simple problem where an observer uses an underlying Hidden Markov Model (HMM) to track a target in one dimension. In this implementation, sensors make noisy observations of the target position at discrete time steps. The tracker learns the transition model for target movement, and the observation model for the noisy sensors, and uses these to estimate the target position by solving the Bayesian recursive equation online. We show the tracking performance of the system and demonstrate how it can learn the observation model, the transition model, and the external distractor (noise) probability interfering with the observations. In the second implementation, referred to as the Bayesian INference in DAG (BIND), we show how inference can be performed in a Directed Acyclic Graph (DAG) using stochastic circuits. We show how these building blocks can be easily implemented using simple digital logic gates. An advantage of the stochastic electronic implementation is that it is robust to certain types of noise, which may become an issue in integrated circuit (IC) technology with feature sizes in the order of tens of nanometers due to their low noise margin, the effect of high-energy cosmic rays and the low supply voltage. In our framework, the flipping of random individual bits would not affect the system performance because information is encoded in a bit stream. PMID:27047326

  7. Joint NASA Ames/Langley Experimental Evaluation of Integrated Air/Ground Operations for En Route Free Maneuvering

    NASA Technical Reports Server (NTRS)

    Barhydt, Richard; Kopardekar, Parimal; Battiste, Vernol; Doble, Nathan; Johnson, Walter; Lee, Paul; Prevot, Thomas; Smith, Nancy

    2005-01-01

    In order to meet the anticipated future demand for air travel, the National Aeronautics and Space Administration (NASA) is investigating a new concept of operations known as Distributed Air-Ground Traffic Management (DAG-TM). Under the En Route Free Maneuvering component of DAG-TM, appropriately equipped autonomous aircraft self separate from other autonomous aircraft and from managed aircraft that continue to fly under today s Instrument Flight Rules (IFR). Controllers provide separation services between IFR aircraft and assign traffic flow management constraints to all aircraft. To address concept feasibility issues pertaining to integrated air/ground operations at various traffic levels, NASA Ames and Langley Research Centers conducted a joint human-in-the-loop experiment. Professional airline pilots and air traffic controllers flew a total of 16 scenarios under four conditions: mixed autonomous/managed operations at three traffic levels and a baseline all-managed condition at the lowest traffic level. These scenarios included en route flights and descents to a terminal area meter fix in airspace modeled after the Dallas Ft. Worth area. Pilots of autonomous aircraft met controller assigned meter fix constraints with high success. Separation violations by subject pilots did not appear to vary with traffic level and were mainly attributable to software errors and procedural lapses. Controller workload was lower for mixed flight conditions, even at higher traffic levels. Pilot workload was deemed acceptable under all conditions. Controllers raised several safety concerns, most of which pertained to the occurrence of near-term conflicts between autonomous and managed aircraft. These issues are being addressed through better compatibility between air and ground systems and refinements to air and ground procedures.

  8. Microscopic Aspects of Silicon-Mediated Rice Resistance to Leaf Scald.

    PubMed

    Araujo, Leonardo; Paschoalino, Rayane Silva; Rodrigues, Fabrício Ávila

    2016-02-01

    This study investigated the effect of silicon (Si) on the potentiation of rice resistance against leaf scald at the microscopic level. Rice plants ('Primavera') were grown in a nutrient solution containing 0 (-Si) or 2 mM (+Si) Si. The foliar Si concentration of the +Si plants (3.6 dag/kg) increased in comparison with the -Si plants (0.3 dag/kg). An X-ray microanalysis revealed that the leaf tissue of +Si plants infected with Microdochium oryzae had higher peaks and deposition of insoluble Si than that of -Si plants. The high foliar Si concentration for the +Si plants reduced the expansion of leaf scald lesions. Scanning electron microscopy revealed that fungal hyphae and appressorium-like structures of M. oryzae were more abundant in the leaf surface of -Si plants relative to +Si plants. At both histopathological and ultrastructural levels, fungal hyphae grew abundantly into the leaf tissue of -Si plants. By contrast, rice cell walls were rarely degraded and fungal hyphae were often surrounded by amorphous granular material in the leaf tissue of +Si plants. Conidiophores emerged from stomata 36 h after fungal penetration, and conidia were noticed inside the leaf tissue of the -Si plants in great abundance. The collective results of the present study showed a high concentration and deposition of Si and a considerable deposition of phenolic-like compounds in the leaf tissue of +Si plants. These results indicate that the potentiation of the phenylpropanoid pathway in these plants supplied with Si was favorable for the increase in rice resistance to leaf scald. PMID:26237696

  9. A possible cellular explanation for the NMR-visible mobile lipid (ML) changes in cultured C6 glioma cells with growth.

    PubMed

    Quintero, MariaRosa; Cabañas, Miquel E; Arús, Carles

    2007-01-01

    The NMR-visible mobile lipid (ML) signals of C6 glioma cells have been monitored at 9.4 and 11.7 T (single pulse and 136 ms echo time) from cell pellets by (1)H NMR spectroscopy. A reproducible behavior with growth has been found. ML signals increase from log phase (4 days of culture) to postconfluence (7 days of culture). This ML behavior is paralleled by the percentage of cells containing epifluorescence detectable Nile Red stained cytosolic droplets (range 23%-60% of cells). The number of positive cells increases after seeding (days 0-1), decreases at log phase (days 2-4), increases again at confluence (day 5) and even further at post-confluence (day 7). C6 cells proliferation arrest induced by growth factors deprivation induces an even higher accumulation of cytosolic droplets (up to 100% of cells) and a large ML increase (up to 21-fold with respect to 4-day log phase cells). When neutral lipid content is quantified by thin-layer chromatography (TLC) on total lipid extracts of C6 cells, no statistically significant change can be detected (in microg/10(8) cells) with growth or growth arrest in major neutral lipid containing species (triacylglycerol, TAG, diacylglycerol, DAG, cholesteryl esters, ChoEst) except for DAG, which decreased in post-confluent, 7-day cells. The apparent discrepancy between NMR, optical microscopy and TLC results can be reconciled if possible biophysical changes in the neutral lipid pool with growth are taken into account. A cellular explanation for the observed results is proposed: the TAG-droplet-size-change hypothesis. PMID:17150408

  10. Metabolomic shifts in Brassica napus lines with enhanced BnPLC2 expression impact their response to low temperature stress and plant pathogens.

    PubMed

    Nokhrina, Kateryna; Ray, Heather; Bock, Cheryl; Georges, Fawzy

    2014-01-01

    Phosphatidylinositol-specific phospholipase C2 (PLC2) is a signaling enzyme with hydrolytic activity against membrane-bound phosphoinositides. It catalyzes the cleavage of phosphatidylinositol(4,5)bisphosphate (PtdIns(4,5)P 2) into two initial second messengers, myo-inositol-1,4,5-trisphosphate (InsP 3) and diacylglycerol (DAG). The former, as well as its fully phosphorylated derivative, myo-inositol-1,2,3,4,5,6-hexakisphosphate (InsP 6), play a major role in calcium signaling events within the cell, while DAG may be used in the regeneration of phospholipids or as a precursor for phosphatidic acid (PA) biosynthesis, an important signaling molecule involved in both biotic and abiotic types of stress tolerance. Overexpression of the gene for Brassica napus phospholipase C2 (BnPLC2) in Brassica napus has been shown to enhance drought tolerance, modulate multiple genes involved in different processes and favorably affect hormonal levels in different tissues. We, therefore, undertook the current study with a view to examining, at the metabolome level, its effect on both abiotic (low temperature) and biotic (stem white rot disease) types of stress in canola. Thus, while transgenic plants exhibited a significant rise in maltose levels and a concomitant elevation in some unsaturated free fatty acids (FFAs), glycerol, and glycerol 3-phosphate under subzero temperatures, they accumulated high levels of raffinose, stachyose and other sugars as well as some flavonoids under acclimatization conditions. Collectively, overexpression of BnPLC2 appears to have triggered different metabolite patterns consistent with its abiotic and, to a limited extent, biotic stress tolerance phenotypes. PMID:24787279

  11. The current deconstruction of paradoxes: one sign of the ongoing methodological "revolution".

    PubMed

    Porta, Miquel; Vineis, Paolo; Bolúmar, Francisco

    2015-10-01

    The current deconstruction of paradoxes is one among several signs that a profound renewal of methods for clinical and epidemiological research is taking place; perhaps for some basic life sciences as well. The new methodological approaches have already deconstructed and explained long puzzling apparent paradoxes, including the (non-existent) benefits of obesity in diabetics, or of smoking in low birth weight. Achievements of the new methods also comprise the elucidation of the causal structure of long-disputed and highly complex questions, as Berkson's bias and Simpson's paradox, and clarifying reasons for deep controversies, as those on estrogens and endometrial cancer, or on adverse effects of hormone replacement therapy. These are signs that the new methods can go deeper and beyond the methods in current use. A major example of a highly relevant idea is: when we condition on a common effect of a pair of variables, then a spurious association between such pair is likely. The implications of these ideas are potentially vast. A substantial number of apparent paradoxes may simply be the result of collider biases, a source of selection bias that is common not just in epidemiologic research, but in many types of research in the health, life, and social sciences. The new approaches develop a new framework of concepts and methods, as collider, instrumental variables, d-separation, backdoor path and, notably, Directed Acyclic Graphs (DAGs). The current theoretical and methodological renewal-or, perhaps, "revolution"-may be changing deeply how clinical and epidemiological research is conceived and performed, how we assess the validity and relevance of findings, and how causal inferences are made. Clinical and basic researchers, among others, should get acquainted with DAGs and related concepts. PMID:26164615

  12. Palmitate induces ER calcium depletion and apoptosis in mouse podocytes subsequent to mitochondrial oxidative stress

    PubMed Central

    Xu, S; Nam, S M; Kim, J-H; Das, R; Choi, S-K; Nguyen, T T; Quan, X; Choi, S J; Chung, C H; Lee, E Y; Lee, I-K; Wiederkehr, A; Wollheim, C B; Cha, S-K; Park, K-S

    2015-01-01

    Pathologic alterations in podocytes lead to failure of an essential component of the glomerular filtration barrier and proteinuria in chronic kidney diseases. Elevated levels of saturated free fatty acid (FFA) are harmful to various tissues, implemented in the progression of diabetes and its complications such as proteinuria in diabetic nephropathy. Here, we investigated the molecular mechanism of palmitate cytotoxicity in cultured mouse podocytes. Incubation with palmitate dose-dependently increased cytosolic and mitochondrial reactive oxygen species, depolarized the mitochondrial membrane potential, impaired ATP synthesis and elicited apoptotic cell death. Palmitate not only evoked mitochondrial fragmentation but also caused marked dilation of the endoplasmic reticulum (ER). Consistently, palmitate upregulated ER stress proteins, oligomerized stromal interaction molecule 1 (STIM1) in the subplasmalemmal ER membrane, abolished the cyclopiazonic acid-induced cytosolic Ca2+ increase due to depletion of luminal ER Ca2+. Palmitate-induced ER Ca2+ depletion and cytotoxicity were blocked by a selective inhibitor of the fatty-acid transporter FAT/CD36. Loss of the ER Ca2+ pool induced by palmitate was reverted by the phospholipase C (PLC) inhibitor edelfosine. Palmitate-dependent activation of PLC was further demonstrated by following cytosolic translocation of the pleckstrin homology domain of PLC in palmitate-treated podocytes. An inhibitor of diacylglycerol (DAG) kinase, which elevates cytosolic DAG, strongly promoted ER Ca2+ depletion by low-dose palmitate. GF109203X, a PKC inhibitor, partially prevented palmitate-induced ER Ca2+ loss. Remarkably, the mitochondrial antioxidant mitoTEMPO inhibited palmitate-induced PLC activation, ER Ca2+ depletion and cytotoxicity. Palmitate elicited cytoskeletal changes in podocytes and increased albumin permeability, which was also blocked by mitoTEMPO. These data suggest that oxidative stress caused by saturated FFA leads to

  13. Bayesian Estimation and Inference Using Stochastic Electronics.

    PubMed

    Thakur, Chetan Singh; Afshar, Saeed; Wang, Runchun M; Hamilton, Tara J; Tapson, Jonathan; van Schaik, André

    2016-01-01

    In this paper, we present the implementation of two types of Bayesian inference problems to demonstrate the potential of building probabilistic algorithms in hardware using single set of building blocks with the ability to perform these computations in real time. The first implementation, referred to as the BEAST (Bayesian Estimation and Stochastic Tracker), demonstrates a simple problem where an observer uses an underlying Hidden Markov Model (HMM) to track a target in one dimension. In this implementation, sensors make noisy observations of the target position at discrete time steps. The tracker learns the transition model for target movement, and the observation model for the noisy sensors, and uses these to estimate the target position by solving the Bayesian recursive equation online. We show the tracking performance of the system and demonstrate how it can learn the observation model, the transition model, and the external distractor (noise) probability interfering with the observations. In the second implementation, referred to as the Bayesian INference in DAG (BIND), we show how inference can be performed in a Directed Acyclic Graph (DAG) using stochastic circuits. We show how these building blocks can be easily implemented using simple digital logic gates. An advantage of the stochastic electronic implementation is that it is robust to certain types of noise, which may become an issue in integrated circuit (IC) technology with feature sizes in the order of tens of nanometers due to their low noise margin, the effect of high-energy cosmic rays and the low supply voltage. In our framework, the flipping of random individual bits would not affect the system performance because information is encoded in a bit stream. PMID:27047326

  14. LA and ALA prevent glucose intolerance in obese male rats without reducing reactive lipid content, but cause tissue-specific changes in fatty acid composition.

    PubMed

    Matravadia, Sarthak; Zabielski, Piotr; Chabowski, Adrian; Mutch, David M; Holloway, Graham P

    2016-04-01

    While the cause of Type 2 diabetes remains poorly defined, the accumulation of reactive lipids within white adipose tissue, skeletal muscle, and liver have been repeatedly implicated as underlying mechanisms. The ability of polyunsaturated fatty acids (PUFAs) to prevent the development of insulin resistance has gained considerable interest in recent years; however, the mechanisms-of-action remain poorly described. Therefore, we determined the efficacy of diets supplemented with either linoleic acid (LA) or α-linolenic acid (ALA) in preventing insulin resistance and reactive lipid accumulation in key metabolic tissues of the obese Zucker rat. Obese Zucker rats displayed impaired glucose homeostasis and reduced n-3 and n-6 PUFA content in the liver and epididymal white adipose tissue (EWAT). After the 12-wk feeding intervention, both LA- and ALA-supplemented diets prevented whole body glucose and insulin intolerance; however, ALA had a more pronounced effect. These changes occurred in association with n-3 and n-6 accumulation in all tissues studied, albeit to different extents (EWAT > liver > muscle). Triacylglycerol (TAG), diacylglycerol (DAG), ceramide, and sphingolipid accumulation were not attenuated in obese animals supplemented with either LA or ALA, suggesting that preservation of glucose homeostasis occurred independent of changes in reactive lipid content. However, PUFA-supplemented diets differentially altered the fatty acid composition of TAGs, DAGs, and PLs in a tissue-specific manner, suggesting essential fatty acid metabolism differs between tissues. Together, our results indicate that remodeling of the fatty acid composition of various lipid fractions may contribute to the improved glucose tolerance observed in obese rats fed PUFA-supplemented diets. PMID:26764053

  15. The exocytotic signaling pathway induced by nerve growth factor in the presence of lyso-phosphatidylserine in rat peritoneal mast cells involves a type D phospholipase.

    PubMed

    Seebeck, J; Westenberger, K; Elgeti, T; Ziegler, A; Schütze, S

    2001-12-15

    Nerve growth factor (NGF) has been previously shown to induce exocytosis in rat peritoneal mast cells (RPMCs) in the presence of lyso-phosphatidylserine (lysoPS) by interacting with high-affinity NGF receptors of the TrkA-type. In RPMCs, type D phosphatidylcholine-selective phospholipases (PLDs) have been postulated to be involved in some exocytotic signaling pathways induced by different agonists. The aim of the present study was to assess a putative functional role of PLD for NGF/lysoPS-induced exocytosis in RPMCs. In 1-[14C]palmitoyl-2-lyso-3-phosphatidylcholine-labelled RPMCs, NGF/lysoPS stimulated the formation of diacylglycerol (DAG) and, in the presence of ethanol (1% [v/v]), phosphatidylethanol (PEtOH). These data indicate PLD-activation by NGF/lysoPS in RPMCs. Preincubation of RPMCs for 2 min with ethanol, an inhibitor of PLD-derived DAG-formation, dose-dependently (IC(50): 0.6% [v/v]) and agonist-selectively inhibited the NGF/lysoPS induced release of [3H]serotonin ([3H]5-HT) in [3H]5-HT-loaded RPMCs, confirming the functional importance of PLD-action. Exocytosis and PEtOH-production was potently inhibited by the broad-spectrum serine/threonine kinase inhibitor staurosporine and activated by the protein kinase C(PKC)-activator PMA (phorbol-12-myristate-13-acetate) suggesting a role for PKC as mediator for NGF/lysoPS-induced activation of PLD. PMID:11730981

  16. Can Sibling Sex Ratios Be Used as a Valid Test for the Prenatal Androgen Hypothesis of Autism Spectrum Disorders?

    PubMed Central

    Cheslack-Postava, Keely; Susser, Ezra; Liu, Kayuet; Bearman, Peter S.

    2015-01-01

    Background Sibling sex ratios have been applied as an indirect test of a hypothesized association between prenatal testosterone levels and risk for autism, a developmental disorder disproportionately affecting males. Differences in sibling sex ratios between those with and without autism would provide evidence of a shared risk factor for autism and offspring sex. Conclusions related to prenatal testosterone, however, require additional assumptions. Here, we used directed acyclic graphs (DAGs) to clarify the elements required for a valid test of the hypothesis that sibling sex ratios differ between children with and without autism. We then conducted such a test using a large, population-based sample of children. Methods Over 1.1 million subjects, born in California from 1992–2007, and identified through birth records, were included. The association between autism diagnosis, determined using the administrative database of the California Department of Developmental Services, and the sex of the subsequent sibling was examined using generalized estimating equations. Sources of potential bias identified using DAGs were addressed. Results Among male children with autism, 52.2% of next-born siblings were brothers, versus 51.0% for unaffected males. For females with autism, 50.2% of following siblings were brothers versus 51.2% among control females. The relative risk of a subsequent male sibling associated with autism diagnosis was 1.02 (95% confidence interval: 0.99, 1.04). Conclusions In a large, population-based sample we failed to find evidence suggesting an excess of brothers among children with autism while controlling for several threats to validity. This test cannot rule out a role of any given exposure, including prenatal testosterone, in either risk of autism or offspring sex ratio, but suggests against a common cause of both. PMID:26495967

  17. Phospholipid biosynthesis in the oyster protozoan parasite, Perkinsus marinus.

    PubMed

    Lund, Eric D; Chu, Fu-Lin E

    2002-05-01

    Perkinsus marinus is a protozoan parasite that causes high mortality in its commercially and ecologically important host, the Eastern oyster Crassostrea virginica. In order to understand the host-parasite relationship in lipid metabolism, the ability of P. marinus to synthesize phospholipids from polar headgroup precursors was investigated. Pulse/chase experiments were conducted using radiolabled serine, choline, ethanolamine and inositol. Timecourse incubations revealed that in vitro cultured P. marinus meronts can utilize the cytidine diphosphate-diacylglycerol (CDP-DAG) pathway to synthesize phosphatidylinositol (PI) from inositol and phosphatidylserine (PS) from serine. Serine label was also incorporated into phosphatidylethanolamine (PE), phosphatidylcholine (PC) and lysophosphatidylcholine (LPC). Incubations of P. marinus cells with increasing concentrations of radiolabeled serine resulted in more radioactivity recovered in neutral lipids than in polar lipids at the highest substrate concentration tested (344 microM). This suggests that excess serine label was being utilized for fatty acid synthesis and stored as triacylglycerols. Additional incubations were conducted with radiolabeled choline and ethanolamine at concentrations equimolar to the highest serine concentration tested. Ethanolamine label was also incorporated into PE, PS, PC and LPC. Choline label was incorporated into PC. These results suggest the presence of three pathways for de novo synthesis of phospholipids in P. marinus: CDP-choline, CDP-ethanolamine and CDP-DAG. At equivalent substrate concentrations (344 microM) the highest incorporation of labeled substrate into total phospholipids was with serine followed by ethanolamine and choline, respectively. P. marinus phospholipid biosynthetic capabilities appear to be similar to those of Plasmodium and Trypanosoma species. PMID:12034458

  18. Effect of a sustained reduction in plasma free fatty acid concentration on insulin signalling and inflammation in skeletal muscle from human subjects

    PubMed Central

    Liang, Hanyu; Tantiwong, Puntip; Sriwijitkamol, Apiradee; Shanmugasundaram, Karthigayan; Mohan, Sumathy; Espinoza, Sara; DeFronzo, Ralph A; Dubé, John J; Musi, Nicolas

    2013-01-01

    Free fatty acids (FFAs) have been implicated in the pathogenesis of insulin resistance. Reducing plasma FFA concentration in obese and type 2 diabetic (T2DM) subjects improves insulin sensitivity. However, the molecular mechanism by which FFA reduction improves insulin sensitivity in human subjects is not fully understood. In the present study, we tested the hypothesis that pharmacological FFA reduction enhances insulin action by reducing local (muscle) inflammation, leading to improved insulin signalling. Insulin-stimulated total glucose disposal (TGD), plasma FFA species, muscle insulin signalling, IκBα protein, c-Jun phosphorylation, inflammatory gene (toll-like receptor 4 and monocyte chemotactic protein 1) expression, and ceramide and diacylglycerol (DAG) content were measured in muscle from a group of obese and T2DM subjects before and after administration of the antilipolytic drug acipimox for 7 days, and the results were compared to lean individuals. We found that obese and T2DM subjects had elevated saturated and unsaturated FFAs in plasma, and acipimox reduced all FFA species. Acipimox-induced reductions in plasma FFAs improved TGD and insulin signalling in obese and T2DM subjects. Acipimox increased IκBα protein (an indication of decreased IκB kinase–nuclear factor κB signalling) in both obese and T2DM subjects, but did not affect c-Jun phosphorylation in any group. Acipimox also decreased inflammatory gene expression, although this reduction only occurred in T2DM subjects. Ceramide and DAG content did not change. To summarize, pharmacological FFA reduction improves insulin signalling in muscle from insulin-resistant subjects. This beneficial effect on insulin action could be related to a decrease in local inflammation. Notably, the improvements in insulin action were more pronounced in T2DM, indicating that these subjects are more susceptible to the toxic effect of FFAs. PMID:23529132

  19. A single prior bout of exercise protects against palmitate-induced insulin resistance despite an increase in total ceramide content.

    PubMed

    Thrush, A Brianne; Harasim, Ewa; Chabowski, Adrian; Gulli, Roberto; Stefanyk, Leslie; Dyck, David J

    2011-05-01

    Ceramide accumulation has been implicated in the impairment of insulin-stimulated glucose transport in skeletal muscle following saturated fatty acid (FA) exposure. Importantly, a single bout of exercise can protect against acute lipid-induced insulin resistance. The mechanism by which exercise protects against lipid-induced insulin resistance is not completely known but may occur through a redirection of FA toward triacylglycerol (TAG) and away from ceramide and diacylglycerol (DAG). Therefore, in the current study, an in vitro preparation was used to examine whether a prior bout of exercise could confer protection against palmitate-induced insulin resistance and whether the pharmacological [50 μM fumonisin B(1) (FB1)] inhibition of ceramide synthesis in the presence of palmitate could mimic the protective effect of exercise. Soleus muscle of sedentary (SED), exercised (EX), and SED in the presence of FB1 (SED+FB1) were incubated with or without 2 mM palmitate for 4 h. This 2-mM palmitate exposure impaired insulin-stimulated glucose transport (-28%, P < 0.01) and significantly increased ceramide, DAG, and TAG accumulation in the SED group (P < 0.05). A single prior bout of exercise prevented the detrimental effects of palmitate on insulin signaling and caused a partial redistribution of FA toward TAG (P < 0.05). However, the net increase in ceramide content in response to palmitate exposure in the EX group was not different compared with SED, despite the maintenance of insulin sensitivity. The incubation of soleus from SED rats with FB1 (SED+FB1) prevented the detrimental effects of palmitate and caused a redirection of FA toward TAG accumulation (P < 0.05). Therefore, this research suggests that although inhibiting ceramide accumulation can prevent the detrimental effects of palmitate, a single prior bout of exercise appears to protect against palmitate-induced insulin resistance, which may be independent of changes in ceramide content. PMID:21325642

  20. Responsiveness of vomeronasal cells to a newt peptide pheromone, sodefrin as monitored by changes of intracellular calcium concentrations.

    PubMed

    Iwata, Takeo; Nakada, Tomoaki; Toyoda, Fumiyo; Yada, Toshihiko; Shioda, Seiji; Kikuyama, Sakae

    2013-07-01

    A peptide pheromone of the red-bellied male newt, sodefrin was tested for its ability to increase intracellular concentrations of Ca(2+) ([Ca(2+)]i) in the dissociated vomeronasal (VN) cells of females by means of calcium imaging system. The pheromone elicited a marked elevation of [Ca(2+)]i in a small population of VN cells from sexually developed females. The population of cells exhibiting sodefrin-induced elevation of [Ca(2+)]i increased concentration-dependently. A pheromone of a different species was ineffective in this respect. The VN cells from non-reproductive females or from reproductive males scarcely responded to sodefrin in terms of elevating [Ca(2+)]i. In the cells from hypophysectomized and ovariectomized females, the sodefrin-inducible increase of [Ca(2+)]i never occurred. The cells from the operated newts supplemented with prolactin and estradiol exhibited [Ca(2+)]i responses to sodefrin with a high incidence. Thus, sex- and hormone-dependency as well as species-specificity of the responsiveness of the VN cells to sodefrin was evidenced at the cellular level. Subsequently, possibility of involvement of phospholipase C (PLC)-inositol 1,4,5-trisphosphate (IP3) and/or PLC-diacylglycerol (DAG)-protein kinase C (PKC) pathways in increasing [Ca(2+)]i in VN cells in response to sodefrin was explored using pharmacological approaches. The results indicated that PLC is involved in generating the Ca(2+) signal in all sodefrin-responsive VN cells, whereas IP3 in approximately 50% of the cells and DAG-PKC in the remaining cells. In the latter case, the increase of [Ca(2+)]i was postulated to be induced by the influx of Ca(2+) through the L-type channel. The significance of the finding is discussed. PMID:23619348

  1. Chang'E-3 data pre-processing system based on scientific workflow

    NASA Astrophysics Data System (ADS)

    tan, xu; liu, jianjun; wang, yuanyuan; yan, wei; zhang, xiaoxia; li, chunlai

    2016-04-01

    The Chang'E-3(CE3) mission have obtained a huge amount of lunar scientific data. Data pre-processing is an important segment of CE3 ground research and application system. With a dramatic increase in the demand of data research and application, Chang'E-3 data pre-processing system(CEDPS) based on scientific workflow is proposed for the purpose of making scientists more flexible and productive by automating data-driven. The system should allow the planning, conduct and control of the data processing procedure with the following possibilities: • describe a data processing task, include:1)define input data/output data, 2)define the data relationship, 3)define the sequence of tasks,4)define the communication between tasks,5)define mathematical formula, 6)define the relationship between task and data. • automatic processing of tasks. Accordingly, Describing a task is the key point whether the system is flexible. We design a workflow designer which is a visual environment for capturing processes as workflows, the three-level model for the workflow designer is discussed:1) The data relationship is established through product tree.2)The process model is constructed based on directed acyclic graph(DAG). Especially, a set of process workflow constructs, including Sequence, Loop, Merge, Fork are compositional one with another.3)To reduce the modeling complexity of the mathematical formulas using DAG, semantic modeling based on MathML is approached. On top of that, we will present how processed the CE3 data with CEDPS.

  2. EDITORIAL: International Workshop on Monte Carlo Techniques in Radiotherapy Delivery and Verification

    NASA Astrophysics Data System (ADS)

    Verhaegen, Frank; Seuntjens, Jan

    2008-03-01

    PhysicsWeb. At McGill we thank the following departments for support: the Cancer Axis of the Research Institute of the McGill University Health Center, the Faculties of Medicine and Science, the Departments of Oncology and Physics and the Medical Physics Unit. The following companies are thanked: TomoTherapy and Standard Imaging. The American Association of Physicists in Medicine and the International Atomic Energy Agency are gratefully acknowledged for endorsing the meeting. A final word of thanks goes out to all of those who contributed to the successful Workshop: first of all our administrative assistant Ms Margery Knewstubb, the website developer Dr François DeBlois, the two heads of the logistics team, Ms Emily Poon and Ms Emily Heath, our local medical physics students and staff, the IOP staff and the authors who shared their new and exciting work with us. Editors: Frank Verhaegen and Jan Seuntjens (McGill University) Associate editors: Luc Beaulieu, Iwan Kawrakow, Tony Popescu and David Rogers

  3. LLE Review Quarterly Report July-September 2000

    SciTech Connect

    2000-12-01

    {sub 2}-filled targets) using current-mode detectors (i.e., many detection events per unit time interval). They show that current-mode detectors can be configured to survey a much larger dynamic range than single-event neutron counters. (4) V. A. Smalyuk, T. R. Boehly, L. S. Iwan, T. J. Kessler, J. P. Knauer, F. J. Marshall, D. D. Meyerhofer, C. Stoeckl, B. Yaakobi, and D. K. Bradley detail a method of measuring the positional dependence of x-ray self-absorption with filtered x-ray framing cameras. They show how compressed shell nonuniformities can be measured by carefully modeling the imaging system. This volume concludes with the LLE's Summer High School Research Program, the FY00 Laser Facility Report, and the National Laser Users' Facility News.

  4. Joint conference of iMEC 2015 (2nd International Manufacturing Engineering Conference & APCOMS 2015 (3rd Asia-Pacific Conference on Manufacturing Systems)

    NASA Astrophysics Data System (ADS)

    2016-02-01

    Properties Synthesized through Top Down Approach”, which was warmly welcomed by an eager and highly motivated audience. The second keynote speaker was Prof. Ir. Dradjad Irianto (ITB, Indonesia) on “Collaborative Manufacturing for Small-medium Enterprises”. The organizers are very grateful to them for supporting the conference and sharing their latest research results with the conference participants. The conference organizers would like to express our sincere gratitude and thanks to the honorary chairman Prof. Dato, Dr. Daing Nasir Ibrahim and Prof. Dr. Wan Azhar Wan Yusoff and organizing committee members of iMEC2015, Assoc. Prof. Dr. A.K. Prasada Rao (chairman), Dr. Ing. Mohd Azmir Mohd Azhari (co-chairman), and all committee members Assoc. Prof. Dr. Ahmad Razlan Yusoff, Assoc. Prof. Dr. Ismed Iskandar, Dr Noraini Mohd Razali, Dr Muhammed Nafis Osman Zahid, Dr Noor Mazni Ismail, Dr Izwan Ismail, Dr Zamzuri Hamedon, Dr Faiz Mohd Turan, Ms Suraya Sulaiman and secretariat Miss Zuryaty, Mrs Wan Norshihah, Mr. Mohd Khairulnazri and Mr Rafilah and also a million thanks to the steering committee from APCOMS Prof. Dr. Kadarsyah Suryadi, Prof. Dr. Bermawi, P. Iskandar, Prof. Dr. Abdul Hakim Halim, Prof. Dr. Dradjad Irianto, Dr. TMA Ari Samadhi, Ir. I Made Dana Tangkas and APCOMS organizing committee Prof. Dr. Abdul Hakim Halim (chair), Dr. Rachmawati Wangsaputra (co-chair) and other committee members Dr. Iwan Inrawan Wiratmadja, Dr. Anas Ma'ruf, Dr. Sukoyo and Dr. Wisnu Aribowo. The editorial board are indebted to all of the reviewers who were willing to spend their precious time in reviewing the papers. Thanks also to all who contributed direct or indirectly in making this conference a success, especially Mr. Mohd Khairulnazri in helping the editors complete the proceedings. Dr. Zamzuri Hamedon Editor in-chief iMEC & APCOMS 2015 Associate Editors 1. Assoc. Prof. Dr. A.K. Prasada Rao 2. Assoc. Prof. Dr. Ahmad Razlan Yusoff 3. Dr. Ing. Mohd Azmir Mohd Azhari 4. Dr. Muhammed

  5. Electrodynamic soil plate oscillator: Modeling nonlinear mesoscopic elastic behavior and hysteresis in nonlinear acoustic landmine detection

    NASA Astrophysics Data System (ADS)

    Korman, M. S.; Duong, D. V.; Kalsbeck, A. E.

    2015-10-01

    significant curvature when the soil particle velocity is relatively higher. An oscillator with hysteresis modeled by a distribution of parallel spring elements each with a different threshold slip condition seems to describe fairly linear backbone curve behavior [W. D. Iwan, Transactions of the ASME, J. of Applied Mech., 33,(1966), 893-900], while a single bilinear hysteresis element describes the backbone curvature results in the experiments reported here [T. K. Caughey, Transactions of the ASME, J. of Applied Mech., 27, (1960), 640-643]. When "off target" resonances have a different backbone curvature than "on the mine" backbone curves, then false alarms may be eliminated due to resonances from the natural soil layering. See [R. A. Guyer, J. TenCate, and P. Johnson, "Hysteresis and the Dynamic Elasticity of Consolidated Granular Materials," Phys. Rev. Lett., 82, 16 (1999), 3280-3283] for recent models of nonlinear mesoscopic behavior.

  6. Regaining Lost Separation in a Piloted Simulation of Autonomous Aircraft Operations

    NASA Technical Reports Server (NTRS)

    Barhydt, Richard; Eischeid, Todd M.; Palmer, Michael T.; Wing, David J.

    2002-01-01

    NASA is currently investigating a new concept of operations for the National Airspace System, designed to improve capacity while maintaining or improving current levels of safety. This concept, known as Distributed Air/Ground Traffic Management (DAG-TM), allows appropriately equipped autonomous aircraft to maneuver freely for flight optimization while resolving conflicts with other traffic and staying out of special use airspace and hazardous weather. While Airborne Separation Assurance System (ASAS) tools would normally allow pilots to resolve conflicts before they become hazardous, evaluation of system performance in sudden, near-term conflicts is needed in order to determine concept feasibility. If an acceptable safety level can be demonstrated in these situations, then operations may be conducted with lower separation minimums. An experiment was conducted in NASA Langley s Air Traffic Operations Lab to address issues associated with resolving near-term conflicts and the potential use of lower separation minimums. Sixteen commercial airline pilots flew a total of 32 traffic scenarios that required them to use prototype ASAS tools to resolve close range pop-up conflicts. Required separation standards were set at either 3 or 5 NM lateral spacing, with 1000 ft vertical separation being used for both cases. Reducing the lateral separation from 5 to 3 NM did not appear to increase operational risk, as indicated by the proximity to the intruder aircraft. Pilots performed better when they followed tactical guidance cues provided by ASAS than when they didn't follow the guidance. As air-air separation concepts are evolved, further studies will consider integration issues between ASAS and existing Airborne Collision Avoidance Systems (ACAS).These types of non-normal events will require the ASAS to provide effective alerts and resolutions prior to the time that an Airborne Collision Avoidance System (ACAS) would give a Resolution Advisory (RA). When an RA is issued, a

  7. Two functionally distinct cholecystokinin receptors show different modes of action on Ca2+ mobilization and phospholipid hydrolysis in isolated rat pancreatic acini. Studies using a new cholecystokinin analog, JMV-180.

    PubMed

    Matozaki, T; Göke, B; Tsunoda, Y; Rodriguez, M; Martinez, J; Williams, J A

    1990-04-15

    A new hepatapeptide cholecystokinin (CCK) analog, JMV-180 (Boc-Tyr(SO3-)-Nle-Gly-Trp-Nle-Asp-2-phenylethylester), acts as an agonist at high affinity CCK receptors on rat pancreatic acini to stimulate amylase release but unlike cholecystokinin octapeptide (CCK8) does not act on low affinity CCK receptors to inhibit amylase release (Galas, M. D., Lignon, M. F., Rodriguez, M., Mendre, C., Fulcrand, P., Laur, J., and Martinez, J. (1988) Am. J. Physiol. 254, G176-G188). To investigate the biochemical mechanisms initiated by CCK acting on each class of CCK receptor, the effects of JMV-180 and CCK8 on amylase release, Ca2+ mobilization, and phospholipid hydrolysis were studied in isolated rat pancreatic acini. When acini were loaded with the intracellular Ca2+ chelator BAPTA, amylase release stimulated by both JMV-180 and CCK8 was reduced. Measurement of 45Ca2+ efflux and cytosolic free calcium concentration ([Ca2+]i) by the fluorescence of fura-2-loaded acini in a stirred cuvette showed that JMV-180 induced a concentration-dependent increase but with a maximal response only two-thirds that induced by CCK8. When [Ca2+]i of individual fura-2-loaded acinar cells was measured by microspectrofluorometry, all concentrations of JMV-180 (1 nM-10 microM) induced repetitive transient [Ca2+]i spikes (Ca2+ oscillations). By contrast, stimulation with a high concentration of CCK8 (1 nM) caused a large increase in [CA2+]i followed by a small sustained elevation of [Ca2+]i. The measurement of inositol trisphosphate (IP3) production by both [3H]inositol labeling and 1,4,5-IP3 radioreceptor assay showed that JMV-180 had only minimal effects at 10 microM in contrast to the large increase induced by high concentrations of CCK8 (more than 1 nM). JMV-180 blocked the effect of a high concentration of CCK8 on both [Ca2+]i and 1,4,5-IP3 productions but did not affect the response to carbamylcholine. JMV-180 caused a delayed monophasic stimulation of 1,2-diacylglycerol (DAG) sustained to 60 min

  8. Signal transduction in esophageal and LES circular muscle contraction.

    PubMed Central

    Harnett, K. M.; Cao, W.; Kim, N.; Sohn, U. D.; Rich, H.; Behar, J.; Biancani, P.

    1999-01-01

    Contraction of normal esophageal circular muscle (ESO) in response to acetylcholine (ACh) is linked to M2 muscarinic receptors activating at least three intracellular phospholipases, i.e., phosphatidylcholine-specific phospholipase C (PC-PLC), phospholipase D (PLD), and the high molecular weight (85 kDa) cytosolic phospholipase A2 (cPLA2) to induce phosphatidylcholine (PC) metabolism, production of diacylglycerol (DAG) and arachidonic acid (AA), resulting in activation of a protein kinase C (PKC)-dependent pathway. In contrast, lower esophageal sphincter (LES) contraction induced by maximally effective doses of ACh is mediated by muscarinic M3 receptors, linked to pertussis toxin-insensitive GTP-binding proteins of the G(q/11) type. They activate phospholipase C, which hydrolyzes phosphatidylinositol bisphosphate (PIP2), producing inositol 1,4,5-trisphosphate (IP3) and DAG. IP3 causes release of intracellular Ca++ and formation of a Ca++-calmodulin complex, resulting in activation of myosin light chain kinase and contraction through a calmodulin-dependent pathway. Signal transduction pathways responsible for maintenance of LES tone are quite distinct from those activated during contraction in response to maximally effective doses of agonists (e.g., ACh). Resting LES tone is associated with activity of a low molecular weight (approximately 14 kDa) pancreatic-like (group 1) secreted phospholipase A2 (sPLA2) and production of arachidonic acid (AA), which is metabolized to prostaglandins and thromboxanes. These AA metabolites act on receptors linked to G-proteins to induce activation of PI- and PC-specific phospholipases, and production of second messengers. Resting LES tone is associated with submaximal PI hydrolysis resulting in submaximal levels of inositol trisphosphate (IP3-induced Ca++ release, and interaction with DAG to activate PKC. In an animal model of acute esophagitis, acid-induced inflammation alters the contractile pathway of ESO and LES. In LES circular

  9. Use of re-esterified palm oils, differing in their acylglycerol structure, in weaning-piglet diets.

    PubMed

    Vilarrasa, E; Barroeta, A C; Tres, A; Esteve-Garcia, E

    2015-08-01

    Re-esterified oils are new fat sources obtained from chemical esterification of acid oils with glycerol (both economically interesting by-products from oil refining and biodiesel industries, respectively). The different fatty acid (FA) positional distribution and acylglycerol composition of re-esterified oils may enhance the apparent absorption of saturated fatty acids (SFA) and, thus, their overall nutritive value. The aim of the present study was to investigate the potential use of re-esterified palm oils, in comparison with their corresponding acid and native oils, and also with an unsaturated fat source in weaning-piglet diets. The parameters assessed were: FA apparent absorption, acylglycerol and free fatty acid (FFA) composition of feces, and growth performance. One-hundred and twenty weaning piglets (average weight of 8.50±1.778 kg) were blocked by initial BW (six blocks) and randomly assigned to five dietary treatments, resulting in four piglets per pen and six replicates per treatment. Dietary treatments were a basal diet supplemented with 10% (as-fed basis) of native soybean oil (SN), native palm oil (PN), acid palm oil (PA), re-esterified palm oil low in mono- (MAG) and diacylglycerols (DAG) (PEL), or re-esterified palm oil high in MAG and DAG (PEH). Results from the digestibility balance showed that SN reached the greatest total FA apparent absorption, and statistically different from PN, PA and PEL (P0.05), but PEH achieved the greatest total FA apparent absorption. Animals fed PEL, despite the fact that PEL oil contained more sn-2 SFA, did not show an improved absorption of SFA (P>0.05). Animals fed PA and PN showed similar apparent absorption coefficients (P>0.05), despite the high FFA content of PA oil. The acylglycerol and FFA composition of feces was mainly composed of FFA. There were no significant differences in growth performance (P>0.05). Results of the present study suggest that, despite the different acylglycerol structure of re

  10. Diacylglycerol kinase epsilon in bovine and rat photoreceptor cells. Light-dependent distribution in photoreceptor cells.

    PubMed

    Natalini, Paola M; Zulian, Sandra E; Ilincheta de Boschero, Mónica G; Giusto, Norma M

    2013-07-01

    The present study shows the selective light-dependent distribution of 1,2-diacylglycerol kinase epsilon (DAGKɛ) in photoreceptor cells from bovine and albino rat retina. Immunofluorescence microscopy in isolated rod outer segments from bleached bovine retinas (BBROS) revealed a higher DAGKɛ signal than that found in rod outer segments from dark-adapted bovine retinas (BDROS). The light-dependent outer segment localization of DAGKɛ was also observed by immunohistochemistry in retinas from albino rats. DAGK activity, measured in terms of phosphatidic acid formation from a) [(3)H]DAG and ATP in the presence of EGTA and R59022, a type I DAGK inhibitor, or b) [γ-(32)P]ATP and 1-stearoyl, 2-arachidonoylglycerol (SAG), was found to be significantly higher in BBROS than in BDROS. Higher light-dependent DAGK activity (condition b) was also found when ROS were isolated from dark-adapted rat retinas exposed to light. Western blot analysis of isolated ROS proteins from bovine and rat retinas confirmed that illumination increases DAGKɛ content in the outer segments of these two species. Light-dependent DAGKɛ localization in the outer segment was not observed when U73122, a phospholipase C inhibitor, was present prior to the exposure of rat eyecups (in situ model) to light. Furthermore, no increased PA synthesis from [(3)H]DAG and ATP was observed in the presence of neomycin prior to the exposure of bovine eyecups to light. Interestingly, when BBROS were pre-phosphorylated with ATP in the presence of 1,2-dioctanoyl sn-glycerol (di-C8) or phorbol dibutyrate (PDBu) as PKC activation conditions, higher DAGK activity was observed than in dephosphorylated controls. Taken together, our findings suggest that the selective distribution of DAGKɛ in photoreceptor cells is a light-dependent mechanism that promotes increased SAG removal and synthesis of 1-stearoyl, 2-arachidonoyl phosphatidic acid in the sensorial portion of this cell, thus demonstrating a novel mechanism of light

  11. Analysis of Aircraft Clusters to Measure Sector-Independent Airspace Congestion

    NASA Technical Reports Server (NTRS)

    Bilimoria, Karl D.; Lee, Hilda Q.

    2005-01-01

    The Distributed Air/Ground Traffic Management (DAG-TM) concept of operations* permits appropriately equipped aircraft to conduct Free Maneuvering operations. These independent aircraft have the freedom to optimize their trajectories in real time according to user preferences; however, they also take on the responsibility to separate themselves from other aircraft while conforming to any local Traffic Flow Management (TFM) constraints imposed by the air traffic service provider (ATSP). Examples of local-TFM constraints include temporal constraints such as a required time of arrival (RTA), as well as spatial constraints such as regions of convective weather, special use airspace, and congested airspace. Under current operations, congested airspace typically refers to a sector(s) that cannot accept additional aircraft due to controller workload limitations; hence Dynamic Density (a metric that is indicative of controller workload) can be used to quantify airspace congestion. However, for Free Maneuvering operations under DAG-TM, an additional metric is needed to quantify the airspace congestion problem from the perspective of independent aircraft. Such a metric would enable the ATSP to prevent independent aircraft from entering any local areas of congestion in which the flight deck based systems and procedures may not be able to ensure separation. This new metric, called Gaggle Density, offers the ATSP a mode of control to regulate normal operations and to ensure safety and stability during rare-normal or off-normal situations (e.g., system failures). It may be difficult to certify Free Maneuvering systems for unrestricted operations, but it may be easier to certify systems and procedures for specified levels of Gaggle Density that could be monitored by the ATSP, and maintained through relatively minor flow-rate (RTA type) restrictions. Since flight deck based separation assurance is airspace independent, the challenge is to measure congestion independent of sector

  12. Oxytocin and parturition: a role for increased myometrial calcium and calcium sensitization?

    PubMed

    Arthur, Patrice; Taggart, Michael J; Mitchell, Bryan F

    2007-01-01

    Preterm birth is associated with the majority of all death and chronic disability related to pregnancy, birth and the neonatal period. The costs to families and to the health care system are enormous. Current approaches to prevent or arrest preterm labour have been unsuccessful. This failure is largely based on our poor understanding of the regulation of the timing and maintenance of parturition. Oxytocin (OT) is the most potent known uterine stimulant. It is produced in the hypothalamus and secreted into the maternal bloodstream. However, OT also is produced within the uterine decidua in late gestation and the concentrations increase around the time of labour onset. The receptor for OT (OTR) is a G-protein coupled receptor linked through G alpha(q/11) to phospholipase C (PLC). Activation of PLC causes increased inositol trisphosphate (IP3) and diacyl glycerol (DAG). IP3 activates specific receptors in the sarcoplasmic reticulum to release Ca2+ into the cytosol. This may induce further influx of Ca2+ from the extracellular space and the increased Ca2+, after binding to calmodulin, activates myosin light chain kinase to phosphorylate myosin light chains (MLC) and cause contraction of the myocyte. DAG activates protein kinase C (PKC), several isoforms of which have been implicated in uterine contraction, but the substrates for this enzyme in the uterine myocyte are essentially unknown. Oxytocin may also cause "Ca2+-sensitization," a process whereby there is a greater contractile force generated from a given increase in cytosolic Ca2+, although the contribution of this process to myometrial contraction remains an area of debate. This phenomenon occurs mainly due to inhibition of myosin light chain phosphatase (MLCP), the enzyme that reverses the phosphorylation of MLC. There are several important potential mediators of this MLCP-inhibitory pathway in the myometrium, including the small monomeric G-protein RhoA, its downstream kinase Rho-associated kinase (ROK). and

  13. Regulated Mucin Secretion from Airway Epithelial Cells

    PubMed Central

    Adler, Kenneth B.; Tuvim, Michael J.; Dickey, Burton F.

    2013-01-01

    Secretory epithelial cells of the proximal airways synthesize and secrete gel-forming polymeric mucins. The secreted mucins adsorb water to form mucus that is propelled by neighboring ciliated cells, providing a mobile barrier which removes inhaled particles and pathogens from the lungs. Several features of the intracellular trafficking of mucins make the airway secretory cell an interesting comparator for the cell biology of regulated exocytosis. Polymeric mucins are exceedingly large molecules (up to 3 × 106 Da per monomer) whose folding and initial polymerization in the ER requires the protein disulfide isomerase Agr2. In the Golgi, mucins further polymerize to form chains and possibly branched networks comprising more than 20 monomers. The large size of mucin polymers imposes constraints on their packaging into transport vesicles along the secretory pathway. Sugar side chains account for >70% of the mass of mucins, and their attachment to the protein core by O-glycosylation occurs in the Golgi. Mature polymeric mucins are stored in large secretory granules ∼1 μm in diameter. These are translocated to the apical membrane to be positioned for exocytosis by cooperative interactions among myristoylated alanine-rich C kinase substrate, cysteine string protein, heat shock protein 70, and the cytoskeleton. Mucin granules undergo exocytic fusion with the plasma membrane at a low basal rate and a high stimulated rate. Both rates are mediated by a regulated exocytic mechanism as indicated by phenotypes in both basal and stimulated secretion in mice lacking Munc13-2, a sensor of the second messengers calcium and diacylglycerol (DAG). Basal secretion is induced by low levels of activation of P2Y2 purinergic and A3 adenosine receptors by extracellular ATP released in paracrine fashion and its metabolite adenosine. Stimulated secretion is induced by high levels of the same ligands, and possibly by inflammatory mediators as well. Activated receptors are coupled to

  14. Lead Abundance In The Martian Mantle Deduced From The Isotopic Data In Snc Meteorites

    NASA Astrophysics Data System (ADS)

    Dreibus, G.; Jagoutz, E.

    Isotopic data are a powerful tool for the study of planetary evolution. Assuming that the SNC meteorites are rocks from Mars their Sm-Nd-, Rb-Sr- and Pb-Pb-isotope systematics reveal the time scale for the chemical evolution of the Martian mantle. From the Rb -Sr isotopic systematic the existence of 3 isotopically distinct reservoirs on Mars was postulated, which remained isolated for a period of 4.3 +/- 0.2 Ga. The basaltic shergottites Shergotty, Zagami and Los Angeles have relatively high radiogenic Sr, which might come from a planetary crust. A second group, characterized by non radiogenic Sr, consists of the two mafic cumulates Nakhla and Chassigny, the olivine rich basaltic shergottites DaG 476, SaU 005, Dhofar 019and the basaltic shergottite QUE 94201, which may represent the depleted mantle. The depletion of this reservoir must have taken place during a very early process. as derived from the primitive Sr isotopes and the existence of Nd-142, the daughter product of the extinct Sm-146, found in Chassigny, the Nakhlites, SaU 005, and DaG476. A third group, with intermediate Sr isotopic composition, represented by the lherzolitic shergottites, could be derived from a primitive, unfractionated mantle. Our observed correlation of Sr-isotopes with Pb-isotopes in SNC's permits to estimate the Pb abundance for the Martian mantle. The Pb isotopes of all measured SNCs show a similar pattern as Sr isotopes. The initial Pb data of Los Angeles, Shergotty, and Zagami from the enriched crustal reservoir and of Nakhla and SaU 005 from the depleted mantle reservoir plot close to the 4.5 Ga Pb -Pb isochron.. We used this correlation to estimate the µ value (238U/204Pb) of 3.1 for the Martian mantle. This corresponds to 366 ppb Pb. Compared to the Earth with a µ = 8.8, Pb is enriched on Mars by at least a fact or of 2.5. The same enrichment was found for all other moderately volatile and volatile elements on Mars. From the high abundance of Pb in the sulfide phases of iron

  15. Geometric phases generated by the non-trivial spatial topology of static vector fields linearly coupled to a neutral spin-endowed particle: application to 171Yb atoms trapped in a 2D optical lattice

    NASA Astrophysics Data System (ADS)

    Bouchiat, Marie-Anne; Bouchiat, Claude

    2012-10-01

    We have constructed the geometric phases emerging from the non-trivial topology of a space-dependent magnetic field B(r), interacting with the spin magnetic moment of a neutral particle. Our basic tool, adapted from a previous work on Berry’s phases, is the space-dependent unitary transformation {U}({\\mathbf {r}}), which leads to the identity, {U}({\\mathbf {r}})^{\\dag }\\, {\\mathbf {S}}\\,{\\bm \\cdot}\\, {\\mathbf {B}}({\\mathbf {r}}) \\, {U}({\\mathbf {r}}) = \\vert {\\mathbf {B}}({\\mathbf {r}}) \\vert \\, S_z, at each point r. In the ‘rotated’ Hamiltonian \\widehat{ H}, \\frac{ \\partial }{\\partial {\\mathbf {r}}} is replaced by the non-Abelian covariant derivative \\frac{ \\partial }{\\partial {\\mathbf {r}}}- \\frac{i}{\\hbar } {A}({\\mathbf {r}}) where {A}({\\mathbf {r}}) = i \\hbar \\, {U}^{\\dag }\\,{\\bm\\cdot}\\, \\frac{ \\partial }{\\partial {\\mathbf {r}}} {U} can be written as A1(r)Sx + A2(r)Sy + A3(r)Sz. The Abelian differentials Ak(r)·dr are given in terms of the Euler angles defining the orientation of B(r). The non-Abelian field {A}({\\mathbf {r}}) transforms as a Yang-Mills field; however, its vanishing ‘curvature’ reveals its purely geometric character. We have defined a perturbation scheme based upon the assumption that in \\widehat{ H} the longitudinal field A3(r) dominates the transverse field A1, 2(r) contributions, evaluated to second order. The geometry embedded in both the vector field A3(r) and the geometric magnetic field \\mathbf { B}_3 ({\\mathbf {r}}) = \\frac{ \\partial }{\\partial {\\mathbf {r}}}\\wedge {{\\mathbf {A}}}_3({\\mathbf {r}}) is described by their associated Aharonov-Bohm phase. As an illustration we study the physics of cold 171Yb atoms dressed by overlaying two circularly polarized stationary waves with orthogonal directions, which form a 2D square optical lattice. The frequency is tuned midway between the two hyperfine levels of the (6s6p)3P1 states to protect the optical B(r) field generated by the

  16. Cross-talk among intracellular signaling pathways mediates the diphenyl ditelluride actions on the hippocampal cytoskeleton of young rats.

    PubMed

    Heimfarth, Luana; Loureiro, Samanta Oliveira; Reis, Karina Pires; de Lima, Bárbara Ortiz; Zamboni, Fernanda; Gandolfi, Talita; Narvaes, Rodrigo; da Rocha, João Batista Teixeira; Pessoa-Pureur, Regina

    2011-10-17

    In the present report, we showed that diphenyl ditelluride (PhTe)(2) induced in vitro hyperphosphorylation of glial fibrillary acidic protein (GFAP), vimentin and neurofilament (NF) subunits in hippocampus of 21 day-old rats. Hyperphosphorylation was dependent on L-voltage dependent Ca(2+) channels (L-VDCC), N-methyl-d-aspartate (NMDA) and metabotropic glutamate receptors, as demonstrated by the specific inhibitors verapamil, DL-AP5 and MCPG, respectively. Also, dantrolene, a ryanodine channel blocker, EGTA and Bapta-AM, extra and intracellular Ca(2+) chelators respectively, totally prevented this effect. Activation of metabotropic glutamate receptors by (PhTe)(2) upregulates phospholipase C (PLC), producing inositol 1, 4, 5-trisphosphate (IP(3)) and diacylglycerol (DAG). Therefore, high Ca(2+) levels and DAG directly activate Ca(2+)/calmodulin-dependent protein kinase (PKCaMII) and protein kinase C (PCK), resulting in the hyperphosphorylation of Ser-57 in the carboxyl-terminal tail domain of the low molecular weight NF subunit (NF-L). Also, the activation of Erk1/2, and p38MAPK resulted in hyperphosphorylation of KSP repeats of the medium molecular weight NF subunit (NF-M). It is noteworthy that PKCaMII and PKC inhibitors prevented (PhTe)(2)-induced Erk1/2MAPK and p38MAPK activation as well as hyperphosphorylation of KSP repeats on NF-M, suggesting that PKCaMII and PKC could be upstream of this activation. Taken together, our results highlight the role of Ca(2+) as a mediator of the (PhTe)(2)-elicited signaling targeting specific phosphorylation sites on IF proteins of neural cells of rat hippocampus. Interestingly, this action shows a significant cross-talk among signaling pathways elicited by (PhTe)(2), connecting glutamate metabotropic cascade with activation of Ca(2+) channels. The extensively phosphorylated amino- and carboxyl- terminal sites could explain, at least in part, the neural dysfunction associated with (PhTe)(2) exposure. PMID:21879721

  17. Efficiency alleles of the Pctr1 modifier locus for plasmacytoma susceptibility.

    PubMed

    Zhang, S L; DuBois, W; Ramsay, E S; Bliskovski, V; Morse, H C; Taddesse-Heath, L; Vass, W C; DePinho, R A; Mock, B A

    2001-01-01

    The susceptibility of BALB/c mice to pristane-induced plasmacytomas is a complex genetic trait involving multiple loci, while DBA/2 and C57BL/6 strains are genetically resistant to the plasmacytomagenic effects of pristane. In this model system for human B-cell neoplasia, one of the BALB/c susceptibility and modifier loci, Pctr1, was mapped to a 5.7-centimorgan (cM) chromosomal region that included Cdkn2a, which encodes p16(INK4a) and p19(ARF), and the coding sequences for the BALB/c p16(INK4a) and p19(ARF) alleles were found to be polymorphic with respect to their resistant Pctr1 counterparts in DBA/2 and C57BL/6 mice (45). In the present study, alleles of Pctr1, Cdkn2a, and D4Mit15 from a resistant strain (BALB/cDAG) carrying DBA/2 chromatin were introgressively backcrossed to the susceptible BALB/c strain. The resultant C.DAG-Pctr1 Cdkn2a D4Mit15 congenic was more resistant to plasmacytomagenesis than BALB/c, thus narrowing Pctr1 to a 1.5-cM interval. Concomitantly, resistant C57BL/6 mice, from which both gene products of the Cdkn2a gene have been eliminated, developed pristane-induced plasma cell tumors over a shorter latency period than the traditionally susceptible BALB/cAn strain. Biological assays of the p16(INK4a) and p19(ARF) alleles from BALB/c and DBA/2 indicated that the BALB/c p16(INK4a) allele was less active than its DBA/2 counterpart in inducing growth arrest of mouse plasmacytoma cell lines and preventing ras-induced transformation of NIH 3T3 cells, while the two p19(ARF) alleles displayed similar potencies in both assays. We propose that the BALB/c susceptibility/modifier locus, Pctr1, is an "efficiency" allele of the p16(INK4a) gene. PMID:11113205

  18. Activation of native TRPC1/C5/C6 channels by endothelin-1 is mediated by both PIP3 and PIP2 in rabbit coronary artery myocytes

    PubMed Central

    Saleh, Sohag N; Albert, Anthony P; Large, William A

    2009-01-01

    We investigate activation mechanisms of native TRPC1/C5/C6 channels (termed TRPC1 channels) by stimulation of endothelin-1 (ET-1) receptor subtypes in freshly dispersed rabbit coronary artery myocytes using single channel recording and immunoprecipitation techniques. ET-1 evoked non-selective cation channel currents with a unitary conductance of 2.6 pS which were not inhibited by either ETA or ETB receptor antagonists, respectively BQ-123 and BQ788, when administered separately. However, in the presence of both antagonists, ET-1-evoked channel activity was abolished indicating that both ETA and ETB receptor stimulation activate this conductance. Stimulation of both ETA and ETB receptors evoked channel activity which was inhibited by the protein kinase C (PKC) inhibitor chelerythrine and by anti-TRPC1 antibodies indicating that activation of both receptor subtypes causes TRPC1 channel activation by a PKC-dependent mechanism. ETA receptor-mediated TRPC1 channel activity was selectively inhibited by phosphoinositol-3-kinase (PI-3-kinase) inhibitors wortmannin (50 nm) and PI-828 and by antibodies raised against phosphoinositol-3,4,5-trisphosphate (PIP3), the product of PI-3-kinase-mediated phosphorylation of phosphatidylinositol 4,5-bisphosphate (PIP2). Moreover, exogenous application of diC8-PIP3 stimulated PKC-dependent TRPC1 channel activity. These results indicate that stimulation of ETA receptors evokes PKC-dependent TRPC1 channel activity through activation of PI-3-kinase and generation of PIP3. In contrast, ETB receptor-mediated TRPC1 channel activity was inhibited by the PI-phospholipase C (PI-PLC) inhibitor U73122. 1-Oleoyl-2-acetyl-sn-glycerol (OAG), an analogue of diacylglycerol (DAG), which is a product of PI-PLC, also activated PKC-dependent TRPC1 channel activity. OAG-induced TRPC1 channel activity was inhibited by anti-phosphoinositol-4,5-bisphosphate (PIP2) antibodies and high concentrations of wortmannin (20 μm) which depleted tissue PIP2 levels. In

  19. Rap1 Activation Plays a Regulatory Role in Pancreatic Amylase Secretion*S⃞

    PubMed Central

    Sabbatini, Maria E.; Chen, Xuequn; Ernst, Stephen A.; Williams, John A.

    2008-01-01

    Rap1 is a member of the Ras superfamily of small GTP-binding proteins and is localized on pancreatic zymogen granules. The current study was designed to determine whether GTP-Rap1 is involved in the regulation of amylase secretion. Rap1A/B and the two Rap1 guanine nucleotide exchange factors, Epac1 and CalDAG-GEF III, were identified in mouse pancreatic acini. A fraction of both Rap1 and Epac1 colocalized with amylase in zymogen granules, but only Rap1 was integral to the zymogen granule membranes. Stimulation with cholecystokinin (CCK), carbachol, and vasoactive intestinal peptide all induced Rap1 activation, as did calcium ionophore A23187, phorbol ester, forskolin, 8-bromo-cyclic AMP, and the Epac-specific cAMP analog 8-pCPT-2′-O-Me-cAMP. The phospholipase C inhibitor U-73122 abolished carbachol- but not forskolin-induced Rap1 activation. Co-stimulation with carbachol and 8-pCPT-2′-O-Me-cAMP led to an additive effect on Rap1 activation, whereas a synergistic effect was seen on amylase release. Although the protein kinase A inhibitor H-89 abolished forskolin-stimulated CREB phosphorylation, it did not modify forskolin-induced GTP-Rap1 levels, excluding PKA participation. Overexpression of Rap1 GTPase-activating protein, which blocked Rap1 activation, reduced the effect of 8-bromo-cyclic AMP, 8-pCPT-2′-O-Me-cAMP, and vasoactive intestinal peptide on amylase release by 60% and reduced CCK- as well as carbachol-stimulated pancreatic amylase release by 40%. These findings indicate that GTP-Rap1 is required for pancreatic amylase release. Rap1 activation not only mediates the cAMP-evoked response via Epac1 but is also involved in CCK- and carbachol-induced amylase release, with their action most likely mediated by CalDAG-GEF III. PMID:18577515

  20. Simulating the reactive transport of nitrogen species in a regional irrigated agricultural groundwater system

    NASA Astrophysics Data System (ADS)

    Bailey, R. T.; Gates, T. K.

    2011-12-01

    The fate and transport of nitrogen (N) species in irrigated agricultural groundwater systems is governed by irrigation patterns, cultivation practices, aquifer-surface water exchanges, and chemical reactions such as oxidation-reduction, volatilization, and sorption, as well as the presence of dissolved oxygen (O2). We present results of applying the newly-developed numerical model RT3D-AG to a 50,400-ha regional study site within the Lower Arkansas River Valley in southeastern Colorado, where elevated concentrations of NO3 have been observed in both groundwater and surface water during the recent decade. Furthermore, NO3 has a strong influence on the fate and transport of other contaminants in the aquifer system such as selenium (Se) through inhibition of reduction of dissolved Se as well as oxidation of precipitate Se from outcropped and bedrock shale. RT3D-AG, developed by appending the multi-species reactive transport finite-difference model RT3D with modular packages that account for variably-saturated transport, the cycling of carbon (C) and N, and the fate and transport of O2 within the soil and aquifer system, simulates organic C and organic N decomposition and mineralization, oxidation-reduction reactions, and sorption. System sources/sinks consist of applied fertilizer and manure; crop uptake of ammonium (NH4) and NO3 during the growing season; mass of O2, NO3, and NH4 associated with irrigation water and canal seepage; mass of O2, NO3, and NH4 transferred to canals and the Arkansas River from the aquifer; and dead root mass and after-harvest stover mass incorporated into the soil organic matter at the end of the growing season. Chemical reactions are simulated using first-order Monod kinetics, wherein the rate of reaction is dependent on the concentration of the reactants as well as temperature and water content of the soil. Fertilizer and manure application timing and loading, mass of seasonal crop uptake, and end-of-season root mass and stover mass are

  1. Exercise-like effects by Estrogen-related receptor-gamma in muscle do not prevent insulin resistance in db/db mice.

    PubMed

    Badin, Pierre-Marie; Vila, Isabelle K; Sopariwala, Danesh H; Yadav, Vikas; Lorca, Sabina; Louche, Katie; Kim, Eun Ran; Tong, Qingchun; Song, Min Sup; Moro, Cedric; Narkar, Vihang A

    2016-01-01

    Dissecting exercise-mimicking pathways that can replicate the benefits of exercise in obesity and diabetes may lead to promising treatments for metabolic disorders. Muscle estrogen-related receptor gamma (ERRγ) is induced by exercise, and when over-expressed in the skeletal muscle mimics exercise by stimulating glycolytic-to-oxidative myofiber switch, mitochondrial biogenesis and angiogenesis in lean mice. The objective of this study was to test whether muscle ERRγ in obese mice mitigates weight gain and insulin resistance. To do so, ERRγ was selectively over-expressed in the skeletal muscle of obese and diabetic db/db mice. Muscle ERRγ over-expression successfully triggered glycolytic-to-oxidative myofiber switch, increased functional mitochondrial content and boosted vascular supply in the db/db mice. Despite aerobic remodeling, ERRγ surprisingly failed to improve whole-body energy expenditure, block muscle accumulation of triglycerides, toxic diacylglycerols (DAG) and ceramides or suppress muscle PKCε sarcolemmal translocation in db/db mice. Consequently, muscle ERRγ did not mitigate impaired muscle insulin signaling or insulin resistance in these mice. In conclusion, obesity and diabetes in db/db mice are not amenable to selective ERRγ-directed programming of classic exercise-like effects in the skeletal muscle. Other biochemical pathways or integrated whole-body effects of exercise may be critical for resisting diabetes and obesity. PMID:27220353

  2. Lipid mediators in diabetic nephropathy

    PubMed Central

    2014-01-01

    The implications of lipid lowering drugs in the treatment of diabetic nephropathy have been considered. At the same time, the clinical efficacy of lipid lowering drugs has resulted in improvement in the cardiovascular functions of chronic kidney disease (CKD) patients with or without diabetes, but no remarkable improvement has been observed in the kidney outcome. Earlier lipid mediators have been shown to cause accumulative effects in diabetic nephropathy (DN). Here, we attempt to analyze the involvement of lipid mediators in DN. The hyperglycemia-induced overproduction of diacyglycerol (DAG) is one of the causes for the activation of protein kinase C (PKCs), which is responsible for the activation of pathways, including the production of VEGF, TGFβ1, PAI-1, NADPH oxidases, and NFҟB signaling, accelerating the development of DN. Additionally, current studies on the role of ceramide are one of the major fields of study in DN. Researchers have reported excessive ceramide formation in the pathobiological conditions of DN. There is less report on the effect of lipid lowering drugs on the reduction of PKC activation and ceramide synthesis. Regulating PKC activation and ceramide biosynthesis could be a protective measure in the therapeutic potential of DN. Lipid lowering drugs also upregulate anti-fibrotic microRNAs, which could hint at the effects of lipid lowering drugs in DN. PMID:25206927

  3. Divergent role of sphingosine 1-phosphate on insulin resistance.

    PubMed

    Fayyaz, Susann; Japtok, Lukasz; Kleuser, Burkhard

    2014-01-01

    Insulin resistance is a complex metabolic disorder in which insulin-sensitive tissues fail to respond to the physiological action of insulin. There is a strong correlation of insulin resistance and the development of type 2 diabetes both reaching epidemic proportions. Dysfunctional lipid metabolism is a hallmark of insulin resistance and a risk factor for several cardiovascular and metabolic disorders. Numerous studies in humans and rodents have shown that insulin resistance is associated with elevations of non-esterified fatty acids (NEFA) in the plasma. Moreover, bioactive lipid intermediates such as diacylglycerol (DAG) and ceramides appear to accumulate in response to NEFA, which may interact with insulin signaling. However, recent work has also indicated that sphingosine 1-phosphate (S1P), a breakdown product of ceramide, modulate insulin signaling in different cell types. In this review, we summarize the current state of knowledge about S1P and insulin signaling in insulin sensitive cells. A specific focus is put on the action of S1P on hepatocytes, pancreatic β-cells and skeletal muscle cells. In particular, modulation of S1P-signaling can be considered as a potential therapeutic target for the treatment of insulin resistance and type 2 diabetes. PMID:24977487

  4. Enhanced effector responses in activated CD8+ T cells deficient in diacylglycerol kinases.

    PubMed

    Riese, Matthew J; Wang, Liang-Chuan S; Moon, Edmund K; Joshi, Rohan P; Ranganathan, Anjana; June, Carl H; Koretzky, Gary A; Albelda, Steven M

    2013-06-15

    Recent clinical trials have shown promise in the use of chimeric antigen receptor (CAR)-transduced T cells; however, augmentation of their activity may broaden their clinical use and improve their efficacy. We hypothesized that because CAR action requires proteins essential for T-cell receptor (TCR) signal transduction, deletion of negative regulators of these signaling pathways would enhance CAR signaling and effector T-cell function. We tested CAR activity and function in T cells that lacked one or both isoforms of diacylglycerol kinase (dgk) expressed highly in T cells, dgkα and dgkζ, enzymes that metabolize the second messenger diacylglycerol (DAG) and limit Ras/ERK activation. We found that primary murine T cells transduced with CARs specific for the human tumor antigen mesothelin showed greatly enhanced cytokine production and cytotoxicity when cocultured with a murine mesothelioma line that stably expresses mesothelin. In addition, we found that dgk-deficient CAR-transduced T cells were more effective in limiting the growth of implanted tumors, both concurrent with and after establishment of tumor. Consistent with our studies in mice, pharmacologic inhibition of dgks also augments function of primary human T cells transduced with CARs. These results suggest that deletion of negative regulators of TCR signaling enhances the activity and function of CAR-expressing T cells and identify dgks as potential targets for improving the clinical potential of CARs. PMID:23576561

  5. Plant phospholipase C family: Regulation and functional role in lipid signaling.

    PubMed

    Singh, Amarjeet; Bhatnagar, Nikita; Pandey, Amita; Pandey, Girdhar K

    2015-08-01

    Phospholipase C (PLC), a major membrane phospholipid hydrolyzing enzyme generates signaling messengers such as diacylglycerol (DAG) and inositol 1,4,5-trisphosphate (IP3) in animals, and their phosphorylated forms such as phosphatidic acid (PA) and inositol hexakisphosphate (IP6) are thought to regulate various cellular processes in plants. Based on substrate specificity, plant PLC family is sub-divided into phosphatidylinositol-PLC (PI-PLC) and phosphatidylcholine-PLC (PC-PLC) groups. The activity of plant PLCs is regulated by various factors and the major ones include, Ca(2+) concentration, phospholipid substrate, post-translational modifications and interacting proteins. Most of the PLC members have been localized at the plasma membrane, suited for their function of membrane lipid hydrolysis. Several PLC members have been implicated in various cellular processes and signaling networks, triggered in response to a number of environmental cues and developmental events in different plant species, which makes them potential candidates for genetically engineering the crop plants for stress tolerance and enhancing the crop productivity. In this review article, we are focusing mainly on the plant PLC signaling and regulation, potential cellular and physiological role in different abiotic and biotic stresses, nutrient deficiency, growth and development. PMID:25933832

  6. Metformin and resveratrol ameliorate muscle insulin resistance through preventing lipolysis and inflammation in hypoxic adipose tissue.

    PubMed

    Zhao, Wenjun; Li, Aiyun; Feng, Xin; Hou, Ting; Liu, Kang; Liu, Baolin; Zhang, Ning

    2016-09-01

    This study aims to investigate the effects of metformin and resveratrol on muscle insulin resistance with emphasis on the regulation of lipolysis in hypoxic adipose tissue. ICR mice were fed with high fat diet (HFD) for 10days with administration of metformin, resveratrol, or intraperitoneal injection of digoxin. Adipose hypoxia, inflammation and cAMP/PKA-dependent lipolysis were investigated. Moreover, lipid deposition and insulin resistance were examined in the muscle. Metformin and resveratrol attenuated adipose hypoxia, inhibited HIF-1α expression and inflammation in the adipose tissue of HFD-fed mice. Metformin and resveratrol inhibited lipolysis through prevention of PKA/HSL activation by decreasing the accumulation of cAMP via preserving PDE3B. Metformin and resveratrol reduced FFAs influx and DAG accumulation, and thus improved insulin signaling in the muscle by inhibiting PKCθ translocation. This study presents a new view of regulating lipid metabolism to ameliorate insulin resistance and provides the clinical guiding significance for obesity and type 2 diabetes with metformin and resveratrol treatment. PMID:27343375

  7. Traffic Behavior Recognition Using the Pachinko Allocation Model

    PubMed Central

    Huynh-The, Thien; Banos, Oresti; Le, Ba-Vui; Bui, Dinh-Mao; Yoon, Yongik; Lee, Sungyoung

    2015-01-01

    CCTV-based behavior recognition systems have gained considerable attention in recent years in the transportation surveillance domain for identifying unusual patterns, such as traffic jams, accidents, dangerous driving and other abnormal behaviors. In this paper, a novel approach for traffic behavior modeling is presented for video-based road surveillance. The proposed system combines the pachinko allocation model (PAM) and support vector machine (SVM) for a hierarchical representation and identification of traffic behavior. A background subtraction technique using Gaussian mixture models (GMMs) and an object tracking mechanism based on Kalman filters are utilized to firstly construct the object trajectories. Then, the sparse features comprising the locations and directions of the moving objects are modeled by PAM into traffic topics, namely activities and behaviors. As a key innovation, PAM captures not only the correlation among the activities, but also among the behaviors based on the arbitrary directed acyclic graph (DAG). The SVM classifier is then utilized on top to train and recognize the traffic activity and behavior. The proposed model shows more flexibility and greater expressive power than the commonly-used latent Dirichlet allocation (LDA) approach, leading to a higher recognition accuracy in the behavior classification. PMID:26151213

  8. The secreted factor Ag1 missing in higher vertebrates regulates fins regeneration in Danio rerio

    PubMed Central

    Ivanova, Anastasiya S.; Shandarin, Igor N.; Ermakova, Galina V.; Minin, Andrey A.; Tereshina, Maria B.; Zaraisky, Andrey G.

    2015-01-01

    Agr family includes three groups of genes, Ag1, Agr2 and Agr3, which encode the thioredoxin domain-containing secreted proteins and have been shown recently to participate in regeneration of the amputated body appendages in amphibians. By contrast, higher vertebrates have only Agr2 and Agr3, but lack Ag1, and have low ability to regenerate the body appendages. Thus, one may hypothesize that loss of Ag1 in evolution could be an important event that led to a decline of the regenerative capacity in higher vertebrates. To test this, we have studied now the expression and role of Ag1 in the regeneration of fins of a representative of another large group of lower vertebrates, the fish Danio rerio. As a result, we have demonstrated that amputation of the Danio fins, like amputation of the body appendages in amphibians, elicits an increase of Ag1 expression in cells of the stump. Furthermore, down-regulation of DAg1 by injections of Vivo-morpholino antisense oligonucleotides resulted in a retardation of the fin regeneration. These data are in a good agreement with the assumption that the loss of Ag1 in higher vertebrates ancestors could lead to the reduction of the regenerative capacity in their modern descendants. PMID:25630240

  9. Convergent, RIC-8-Dependent Gα Signaling Pathways in the Caenorhabditis elegans Synaptic Signaling Network

    PubMed Central

    Reynolds, Nicole K.; Schade, Michael A.; Miller, Kenneth G.

    2005-01-01

    We used gain-of-function and null synaptic signaling network mutants to investigate the relationship of the Gαq and Gαs pathways to synaptic vesicle priming and to each other. Genetic epistasis studies using Gαq gain-of-function and null mutations, along with a mutation that blocks synaptic vesicle priming and the synaptic vesicle priming stimulator phorbol ester, suggest that the Gαq pathway generates the core, obligatory signals for synaptic vesicle priming. In contrast, the Gαs pathway is not required for the core priming function, because steady-state levels of neurotransmitter release are not significantly altered in animals lacking a neuronal Gαs pathway, even though these animals are strongly paralyzed as a result of functional (nondevelopmental) defects. However, our genetic analysis indicates that these two functionally distinct pathways converge and that they do so downstream of DAG production. Further linking the two pathways, our epistasis analysis of a ric-8 null mutant suggests that RIC-8 (a receptor-independent Gα guanine nucleotide exchange factor) is required to maintain both the Gαq vesicle priming pathway and the neuronal Gαs pathway in a functional state. We propose that the neuronal Gαs pathway transduces critical positional information onto the core Gαq pathway to stabilize the priming of selected synapses that are optimal for locomotion. PMID:15489511

  10. Assay and Inhibition of the Purified Catalytic Domain of Diacylglycerol Lipase Beta.

    PubMed

    Singh, Praveen K; Markwick, Rachel; Lu, Leanne; Howell, Fiona V; Williams, Gareth; Doherty, Patrick

    2016-05-17

    The diacylglycerol lipases (DAGLα and DAGLβ) hydrolyze DAG to generate 2-arachidonoylglycerol (2-AG), the principal endocannabinoid and main precursor of arachidonic acid (AA). The DAGLs make distinct tissue specific contributions toward 2-AG and AA levels, and therefore, selective modulators for these enzymes could play crucial roles toward harnessing their therapeutic potential. Relatively high-throughput assays have recently been reported for DAGLα and have proven useful toward the characterization of inhibitors of this enzyme. Similar assays are also warranted for DAGLβ which was the aim of this study. We first adapted previously reported DAGLα membrane assays (using PNPB and DiFMUO as substrates) to measure recombinant DAGLβ activity in membranes. In contrast to results with DAGLα, both substrates provided a relatively limited signal window for measuring DAGLβ activity, however, an improved window was obtained when employing a third commercially available substrate, EnzChek. In order to further improve on the assay parameters, we successfully purified the glutathione S-transferase (GST) tagged catalytic domain of DAGLβ. Activity of the enzyme was confirmed using EnzChek as well as two DAGL inhibitors (THL and OMDM-188). The purified DAGLβ catalytic domain assay described here provides the basis for a relatively clean and convenient assay with the potential to be adapted for high-throughput drug discovery efforts. PMID:27115711

  11. The lipid profile of brown adipose tissue is sex-specific in mice.

    PubMed

    Hoene, Miriam; Li, Jia; Häring, Hans-Ulrich; Weigert, Cora; Xu, Guowang; Lehmann, Rainer

    2014-10-01

    Brown adipose tissue (BAT) is a thermogenic organ with a vital function in small mammals and potential as metabolic drug target in humans. By using high-resolution LC-tandem-mass spectrometry, we quantified 329 lipid species from 17 (sub)classes and identified the fatty acid composition of all phospholipids from BAT and subcutaneous and gonadal white adipose tissue (WAT) from female and male mice. Phospholipids and free fatty acids were higher in BAT, while DAG and TAG levels were higher in WAT. A set of phospholipids dominated by the residue docosahexaenoic acid, which influences membrane fluidity, showed the highest specificity for BAT. We additionally detected major sex-specific differences between the BAT lipid profiles, while samples from the different WAT depots were comparatively similar. Female BAT contained less triacylglycerol and more phospholipids rich in arachidonic and stearic acid whereas another set of fatty acid residues that included linoleic and palmitic acid prevailed in males. These differences in phospholipid fatty acid composition could greatly affect mitochondrial membranes and other cellular organelles and thereby regulate the function of BAT in a sex-specific manner. PMID:25128765

  12. Effect of phorbol derivatives and staurosporine on gravitropic response of primary root of maize

    SciTech Connect

    Mulkey, T.J.; Kim, S.Y. ); Lee, J.S. )

    1991-05-01

    Time-lapse videography and computer-based, video image digitization were used to examine the effects of phorbol derivatives (phorbol 12-myristate 13-acetate, TPA; phorbol 12-myristate 13-acetate 4-O-methyl ether, mTPA) and staurosporine on the kinetics of gravicurvature of primary roots of maize (Zea mays L., Pioneer 3343 and Golden Cross Bantam). Pretreatment of roots with TPA (3 hr, 1 {mu}M) decreases the time lag prior to induction of positive gravicurvature in horizontally-oriented roots by > 60%. The rate of curvature is not significantly different than the rate observed in control roots. Wrongway curvature which is observed in 30-40% of control roots is not observed in TPA-pretreated roots. Oscillatory movements observed in control roots after completion of gravitropic reorientation is completely dampened in TPA-pretreated roots. Pretreatment of roots with mTPA(3hr,1{mu}M), the inactive analog of TPA, does not significantly alter the kinetics of gravicurvature of primary roots of maize. Staurosporine (10{sup {minus}8}M), a microbial alkaloid which has been reported to have antifungal activity and to inhibit phospholipid/Ca{sup ++} dependent protein kinase, completely inhibits TPA-induced alteration of the kinetics of gravitropism. DAG (1-oleoyl-2-acetyl-rac-glycerol), a synthetic diglyceride activator of protein kinase C, exhibits similar activity to TPA. TPA-induced alterations in tissue response to auxin are presented.

  13. Effects of acarbose on fecal nutrients, colonic pH, and short-chain fatty acids and rectal proliferative indices.

    PubMed

    Holt, P R; Atillasoy, E; Lindenbaum, J; Ho, S B; Lupton, J R; McMahon, D; Moss, S F

    1996-09-01

    Acarbose, an alpha-glycosidase inhibitor, treats diabetes mellitus by delaying the digestion and intestinal absorption of dietary carbohydrates. In effective doses, acarbose induces some passage of carbohydrates into the colon. The effect of such chronic carbohydrate transfer on colonic structure and function is unknown. We studied the effects of 1 year of acarbose administration in diabetes mellitus on fecal energy, protein, and fat, including short-chain fatty acids (SCFA) output, fecal pH, and several metabolizing bacterial species. Changes in colonic histology and epithelial cell proliferation were investigated in rectal biopsies. Fecal macronutrient output was unaffected by acarbose, but pH decreased and total SCFA, butyrate, and acetate output were markedly greater. Breath hydrogen output increased after acarbose, but digoxin-metabolizing bacteria and diacylglycerol (DAG) production were unaltered. Compared with the control, acarbose did not induce hyperplasia or change rectal proliferation. However, total fecal SCFA and butyrate output correlated inversely with proliferation in the rectal upper crypt-a biomarker of risk for colonic neoplasia. In conclusion, long-term acarbose administration does not adversely affect colonic function or fecal nutrient output. If increased fecal SCFA and butyrate reduces upper-crypt proliferation, then acarbose may reduce the risk of colonic neoplasia. PMID:8781308

  14. A traditional herbal medicine, rikkunshi-to (TJ-43), prevents intracellular signaling disorders in gastric smooth muscle of diabetic rats.

    PubMed

    Sakai, Yasushi; Nobe, Koji; Maruyama, Yoshiaki; Momose, Kazutaka; Homma, Ikuo

    2004-01-01

    Prevention of diabetic gastrointestinal dysfunction is of utmost importance. The present study demonstrated that diacylglycerol kinase (DGK) activity in diabetic gastric smooth muscle in the resting state was approximately 3.5-fold greater than that in controls. However, oral administration of TJ-43 (1% of food intake) or subcutaneous insulin injection (12 units/kg/day) in streptozotocin-induced diabetic rats (DM) for 2 weeks prevented DGK abnormalities based on the control level. Increased DGK activity in the resting state of DM was inhibited significantly by R59022, neomycin or staurosporine; in contrast, these drugs did not affect DGK activity in controls, insulin-treated DM or TJ-43-treated DM. In controls, the endogenous phosphatidic acid (PA) level was inhibited significantly by R59022 or neomycin but not affected by staurosporine. On the other hand, these three drugs significantly inhibited endogenous PA levels in DM, and neomycin significantly inhibited endogenous PA levels in insulin-treated and TJ-43-treated DM. This suggests that TJ-43 could prevent alteration of DGK activity and PA formation without reduction of blood glucose levels. Moreover, these effects were greater than those of insulin treatment. Results suggested that TJ-43 treatment influenced the hyperreactivity of DGK and DAG formation via phospholipase C activity. In conclusion, TJ-43 can be recommended with respect to enhancement of the quality of life in patients displaying diabetic gastrointestinal complications. PMID:15315262

  15. Characterization of the Mouse and Human Monoacylglycerol O-Acyltransferase 1 (Mogat1) Promoter in Human Kidney Proximal Tubule and Rat Liver Cells.

    PubMed

    Sankella, Shireesha; Garg, Abhimanyu; Agarwal, Anil K

    2016-01-01

    Monoacylglycerol acyltransferase 1 (Mogat1) catalyzes the conversion of monoacylglycerols (MAG) to diacylglycerols (DAG), the precursor of several physiologically important lipids such as phosphatidylcholine, phosphatidylethanolamine and triacylglycerol (TAG). Expression of Mogat1 is tissue restricted and it is highly expressed in the kidney, stomach and adipose tissue but minimally in the normal adult liver. To understand the transcriptional regulation of Mogat1, we characterized the mouse and human Mogat1 promoters in human kidney proximal tubule-2 (HK-2) cells. In-silico analysis revealed several peroxisome proliferator response element (PPRE) binding sites in the promoters of both human and mouse Mogat1. These sites responded to all three peroxisome proliferator activated receptor (PPAR) isoforms such that their respective agonist or antagonist activated or inhibited the expression of Mogat1. PPRE site mutagenesis revealed that sites located at -592 and -2518 are very effective in decreasing luciferase reporter gene activity. Chromatin immunoprecipitation (ChIP) assay using PPARα antibody further confirmed the occupancy of these sites by PPARα. While these assays revealed the core promoter elements necessary for Mogat1 expression, there are additional elements required to regulate its tissue specific expression. Chromosome conformation capture (3C) assay revealed additional cis-elements located ~10-15 kb upstream which interact with the core promoter. These chromosomal regions are responsive to both PPARα agonist and antagonist. PMID:27611931

  16. The BRENDA Tissue Ontology (BTO): the first all-integrating ontology of all organisms for enzyme sources.

    PubMed

    Gremse, Marion; Chang, Antje; Schomburg, Ida; Grote, Andreas; Scheer, Maurice; Ebeling, Christian; Schomburg, Dietmar

    2011-01-01

    BTO, the BRENDA Tissue Ontology (http://www.BTO.brenda-enzymes.org) represents a comprehensive structured encyclopedia of tissue terms. The project started in 2003 to create a connection between the enzyme data collection of the BRENDA enzyme database and a structured network of source tissues and cell types. Currently, BTO contains more than 4600 different anatomical structures, tissues, cell types and cell lines, classified under generic categories corresponding to the rules and formats of the Gene Ontology Consortium and organized as a directed acyclic graph (DAG). Most of the terms are endowed with comments on their derivation or definitions. The content of the ontology is constantly curated with ∼1000 new terms each year. Four different types of relationships between the terms are implemented. A versatile web interface with several search and navigation functionalities allows convenient online access to the BTO and to the enzymes isolated from the tissues. Important areas of applications of the BTO terms are the detection of enzymes in tissues and the provision of a solid basis for text-mining approaches in this field. It is widely used by lab scientists, curators of genomic and biochemical databases and bioinformaticians. The BTO is freely available at http://www.obofoundry.org. PMID:21030441

  17. Scheduling Multilevel Deadline-Constrained Scientific Workflows on Clouds Based on Cost Optimization

    DOE PAGESBeta

    Malawski, Maciej; Figiela, Kamil; Bubak, Marian; Deelman, Ewa; Nabrzyski, Jarek

    2015-01-01

    This paper presents a cost optimization model for scheduling scientific workflows on IaaS clouds such as Amazon EC2 or RackSpace. We assume multiple IaaS clouds with heterogeneous virtual machine instances, with limited number of instances per cloud and hourly billing. Input and output data are stored on a cloud object store such as Amazon S3. Applications are scientific workflows modeled as DAGs as in the Pegasus Workflow Management System. We assume that tasks in the workflows are grouped into levels of identical tasks. Our model is specified using mathematical programming languages (AMPL and CMPL) and allows us to minimize themore » cost of workflow execution under deadline constraints. We present results obtained using our model and the benchmark workflows representing real scientific applications in a variety of domains. The data used for evaluation come from the synthetic workflows and from general purpose cloud benchmarks, as well as from the data measured in our own experiments with Montage, an astronomical application, executed on Amazon EC2 cloud. We indicate how this model can be used for scenarios that require resource planning for scientific workflows and their ensembles.« less

  18. Genomic Analysis of Genes Involved in the Biosynthesis of Very Long Chain Polyunsaturated Fatty Acids in Thraustochytrium sp. 26185.

    PubMed

    Zhao, Xianming; Dauenpen, Meesapyodsuk; Qu, Cunmin; Qiu, Xiao

    2016-09-01

    Thraustochytrium sp. 26185 is a marine protist that can produce a large amount of docosahexaenoic acid (DHA, 22:6n-3), an ω3 very long chain polyunsaturated fatty acid (VLCPUFA) of nutritional importance. However, the mechanism of how this fatty acid is synthesized and assembled into the storage lipid triacylglycerol is unclear. Here we report sequencing of the whole genome and genomic analysis of genes involved in the biosynthesis and assembly of the fatty acids in this species. Genome sequencing produced a total of 2,418,734,139 bp clean sequences with about 62 fold genome coverage. Annotation of the genome sequences revealed 10,797 coding genes. Among them, 10,216 genes could be assigned into 25 KOG classes where 451 genes were specifically assigned to the group of lipid transport and metabolism. Detailed analysis of these genes revealed co-existence of both aerobic pathway and anaerobic pathways for the biosynthesis of DHA in this species. However, in the aerobic pathway, a key gene encoding stearate Δ9 desaturase introducing the first double bond to long chain saturated fatty acid 18:0 was missing from the genome. Genomic survey of genes involved in the acyl trafficking among glycerolipids showed that, unlike plants, this protist did not possess phosphatidylcholine:diacylglycerol cholinephosphotransferase, an important enzyme in bridging two types of glycerolipids, diacylglycerols (DAG) and phosphatidylcholines (PtdCho). These results shed new insight on the biosynthesis and assembly of VLCPUFA in the Thraustochytrium. PMID:27514858

  19. Swedish Experiences In The Application Of Thermography For Establishing Priorities And Quality Control In Residential Dwelling Retrofits

    NASA Astrophysics Data System (ADS)

    Axen, Bengt; Holmsten, Dag

    1981-01-01

    The oldest and largest building thermography consultant in Sweden, Riksbyggen, is also an innovative building developer and contractor. By using thermography not only on their own projects, but also offering a wide variety of thermographic services, Riksbyggen has probably acquired more experiences than any other single inspection unit in the world. As a response to the increasing need for a cost-efficient technique to make energy audits on existing buildings as well as to control the workmanship in retrofits made, Bengt Axen at Riksbyggen has developed a unique building inspection program that efficiently utilizes thermography without compromising the benefits of the technique. As an observer from the outside, the co-author, Dag Holmsten of AGA Corporation, has discussed improving inspection efficiency with Bengt Axen on several occasions and has met with the people invloved in the first test of the "Sundsvall model" in northern Sweden. He gives the institutional background to this new and very practical approach to energy auditing and shows the urgent need for such a technique with global applicability in order to avoid expensive misinvestments of energy conservation funds.

  20. Neutral Lipid Metabolism Influences Phospholipid Synthesis and Deacylation in Saccharomyces cerevisiae

    PubMed Central

    Mora, Gabriel; Scharnewski, Michael; Fulda, Martin

    2012-01-01

    Establishment and maintenance of equilibrium in the fatty acid (FA) composition of phospholipids (PL) requires both regulation of the substrate available for PL synthesis (the acyl-CoA pool) and extensive PL turnover and acyl editing. In the present study, we utilize acyl-CoA synthetase (ACS) deficient cells, unable to recycle FA derived from lipid deacylation, to evaluate the role of several enzymatic activities in FA trafficking and PL homeostasis in Saccharomyces cerevisiae. The data presented show that phospholipases B are not contributing to constitutive PL deacylation and are therefore unlikely to be involved in PL remodeling. In contrast, the enzymes of neutral lipid (NL) synthesis and mobilization are central mediators of FA trafficking. The phospholipid:DAG acyltransferase (PDAT) Lro1p has a substantial effect on FA release and on PL equilibrium, emerging as an important mediator in PL remodeling. The acyl-CoA dependent biosynthetic activities of NL metabolism are also involved in PL homeostasis through active modulation of the substrate available for PL synthesis. In addition TAG mobilization makes an important contribution, especially in cells from stationary phase, to FA availability. Beyond its well-established role in the formation of a storage pool, NL metabolism could play a crucial role as a mechanism to uncouple the pools of PL and acyl-CoAs from each other and thereby to allow independent regulation of each one. PMID:23139841

  1. Maintenance or Collapse: Responses of Extraplastidic Membrane Lipid Composition to Desiccation in the Resurrection Plant Paraisometrum mileense

    PubMed Central

    Yu, Buzhu; Yu, Xiaomei; Li, Weiqi

    2014-01-01

    Resurrection plants usually grow in specific or extreme habitats and have the capacity to survive almost complete water loss. We characterized the physiological and biochemical responses of Paraisometrum mileense to extreme desiccation and found that it is a resurrection plant. We profiled the changes in lipid molecular species during dehydration and rehydration in P. mileense, and compared these with corresponding changes in the desiccation-sensitive plant Arabidopsis thaliana. One day of desiccation was lethal for A. thaliana but not for P. mileense. After desiccation and subsequent rewatering, A. thaliana showed dramatic lipid degradation accompanied by large increases in levels of phosphatidic acid (PA) and diacylglycerol (DAG). In contrast, desiccation and rewatering of P. mileense significantly decreased the level of monogalactosyldiacylglycerol and increased the unsaturation of membrane lipids, without changing the level of extraplastidic lipids. Lethal desiccation in P. mileense caused massive lipid degradation, whereas the PA content remained at a low level similar to that of fresh leaves. Neither damage nor repair processes, nor increases in PA, occurred during non-lethal desiccation in P. mileense. The activity of phospholipase D, the main source of PA, was much lower in P. mileense than in A. thaliana under control conditions, or after either dehydration or rehydration. It was demonstrated that low rates of phospholipase D-mediated PA formation in P. mileense might limit its ability to degrade lipids to PA, thereby maintaining membrane integrity following desiccation. PMID:25068901

  2. Gene expression profiling gut microbiota in different races of humans.

    PubMed

    Chen, Lei; Zhang, Yu-Hang; Huang, Tao; Cai, Yu-Dong

    2016-01-01

    The gut microbiome is shaped and modified by the polymorphisms of microorganisms in the intestinal tract. Its composition shows strong individual specificity and may play a crucial role in the human digestive system and metabolism. Several factors can affect the composition of the gut microbiome, such as eating habits, living environment, and antibiotic usage. Thus, various races are characterized by different gut microbiome characteristics. In this present study, we studied the gut microbiomes of three different races, including individuals of Asian, European and American races. The gut microbiome and the expression levels of gut microbiome genes were analyzed in these individuals. Advanced feature selection methods (minimum redundancy maximum relevance and incremental feature selection) and four machine-learning algorithms (random forest, nearest neighbor algorithm, sequential minimal optimization, Dagging) were employed to capture key differentially expressed genes. As a result, sequential minimal optimization was found to yield the best performance using the 454 genes, which could effectively distinguish the gut microbiomes of different races. Our analyses of extracted genes support the widely accepted hypotheses that eating habits, living environments and metabolic levels in different races can influence the characteristics of the gut microbiome. PMID:26975620

  3. Lipid composition and metabolism in embryos of Brassica napus

    SciTech Connect

    Sparace, S.A. ); Pomroy, M.K. )

    1990-05-01

    Seven and 14-day old microspore-derived developing embryos of the low-erucate Brassica napus L. (cv. Topas) were analyzed for their acyl lipid composition and capacity to incorporate ({sup 14}C)acetate into lipid. The most significant changes in the lipid compositions of these ages of embryos are (1) increased total lipid from 2 to 5% of fresh weight; (2) increased proportion of TAG from 31 to 74%, and shifts in the fatty acid composition of TAG from 25 to 50% 18:1; 28 to 23% 18:2; and 24 to 13% 18:3. Lipids of 7-day embryos also consist of primarily 8% DAG, 2% MG, 12% FFA, 10% DGDG, 15% PA and approximately 5% each of PC, PE and PG. The levels of these lipids generally decrease as the embryos mature and accumulate TAG. ({sup 14}C)Acetate is incorporated into all lipids and fatty acids except 18:2 or 18:3. As much as 39, 59 and 34% of the fatty acid radioactivity of Mg was recovered in 20:0, 22:0 and 24:0, respectively.

  4. A Novel Pathway for Triacylglycerol Biosynthesis Is Responsible for the Accumulation of Massive Quantities of Glycerolipids in the Surface Wax of Bayberry (Myrica pensylvanica) Fruit[OPEN

    PubMed Central

    Ohlrogge, John B.

    2016-01-01

    Bayberry (Myrica pensylvanica) fruits synthesize an extremely thick and unusual layer of crystalline surface wax that accumulates to 32% of fruit dry weight, the highest reported surface lipid accumulation in plants. The composition is also striking, consisting of completely saturated triacylglycerol, diacylglycerol, and monoacylglycerol with palmitate and myristate acyl chains. To gain insight into the unique properties of Bayberry wax synthesis, we examined the chemical and morphological development of the wax layer, monitored wax biosynthesis through [14C]-radiolabeling, and sequenced the transcriptome. Radiolabeling identified sn-2 monoacylglycerol as an initial glycerolipid intermediate. The kinetics of [14C]-DAG and [14C]-TAG accumulation and the regiospecificity of their [14C]-acyl chains indicated distinct pools of acyl donors and that final TAG assembly occurs outside of cells. The most highly expressed lipid-related genes were associated with production of cutin, whereas transcripts for conventional TAG synthesis were >50-fold less abundant. The biochemical and expression data together indicate that Bayberry surface glycerolipids are synthesized by a pathway for TAG synthesis that is related to cutin biosynthesis. The combination of a unique surface wax and massive accumulation may aid understanding of how plants produce and secrete non-membrane glycerolipids and also how to engineer alternative pathways for lipid production in non-seeds. PMID:26744217

  5. Nod factor-induced phosphatidic acid and diacylglycerol pyrophosphate formation: a role for phospholipase C and D in root hair deformation.

    PubMed

    den Hartog, M; Musgrave, A; Munnik, T

    2001-01-01

    Rhizobium-secreted nodulation factors are lipochitooligosaccharides that trigger the initiation of nodule formation on host legume roots. The first visible effect is root hair deformation, but the perception and signalling mechanisms that lead to this response are still unclear. When we treated Vicia sativa seedlings with mastoparan root hairs deformed, suggesting that G proteins are involved. To investigate whether mastoparan and Nod factor activate lipid signalling pathways initiated by phospholipase C (PLC) and D (PLD), seedlings were radiolabelled with [(32)P]orthophosphate prior to treatment. Mastoparan stimulated increases in phosphatidic acid (PA) and diacylglycerol pyrophosphate, indicative of PLD or PLC activity in combination with diacylglycerol kinase (DGK) and PA kinase. Treatment with Nod factor had similar effects, although less pronounced. The inactive mastoparan analogue Mas17 had no effect. The increase in PA was partially caused by the activation of PLD that was monitored by its in vivo transphosphatidylation activity. The application of primary butyl alcohols, inhibitors of PLD activity, blocked root hair deformation. Using different labelling strategies, evidence was provided for the activation of DGK. Since the PLC antagonist neomycin inhibited root hair deformation and the formation of PA, we propose that PLC activation produced diacylglycerol (DAG), which was subsequently converted to PA by DGK. The roles of PLC and PLD in Nod factor signalling are discussed. PMID:11169182

  6. The PKC-NFκB Signaling Pathway Induces APOBEC3B Expression in Multiple Human Cancers

    PubMed Central

    Leonard, Brandon; McCann, Jennifer L.; Starrett, Gabriel J.; Kosyakovsky, Leah; Luengas, Elizabeth M.; Molan, Amy M.; Burns, Michael B.; McDougle, Rebecca M.; Parker, Peter J.; Brown, William L.; Harris, Reuben S.

    2015-01-01

    Overexpression of the antiviral DNA cytosine deaminase APOBEC3B has been linked to somatic mutagenesis in many cancers. HPV infection accounts for APOBEC3B upregulation in cervical and head/neck cancers, but the mechanisms underlying non-viral malignancies are unclear. In this study, we investigated the signal transduction pathways responsible for APOBEC3B upregulation. Activation of protein kinase C (PKC) by the diacylglycerol (DAG) mimic phorbol-myristic acid (PMA) resulted in specific and dose-responsive increases in APOBEC3B expression and activity, which could then be strongly suppressed by PKC or NFκB inhibition. PKC activation caused the recruitment of RELB, but not RELA, to the APOBEC3B promoter implicating non-canonical NFκB signaling. Notably, PKC was required for APOBEC3B upregulation in cancer cell lines derived from multiple tumor types. By revealing how APOBEC3B is upregulated in many cancers, our findings suggest that PKC and NFκB inhibitors may be repositioned to suppress cancer mutagenesis, dampen tumor evolution, and decrease the probability of adverse outcomes such as drug resistance and metastases. PMID:26420215

  7. Oil is on the agenda: Lipid turnover in higher plants.

    PubMed

    Kelly, Amélie A; Feussner, Ivo

    2016-09-01

    Lipases hydrolyze ester bonds within lipids. This process is called lipolysis. They are key players in lipid turnover and involved in numerous metabolic pathways, many of which are shared between organisms like the mobilization of neutral or storage lipids or lipase-mediated membrane lipid homeostasis. Some reactions though are predominantly present in certain organisms, such as the production of signaling molecules (endocannabinoids) by diacylglycerol (DAG) and monoacylglycerol (MAG) lipases in mammals and plants or the jasmonate production in flowering plants. This review aims at giving an overview of the different functional classes of lipases and respective well-known activities, with a focus on the most recent findings in plant biology for selected classes. Here we will put an emphasis on the physiological role and contribution of lipases to the turnover of neutral lipids found in seed oil and other vegetative tissue as candidates for increasing the economical values of crop plants. This article is part of a Special Issue entitled: Plant Lipid Biology edited by Kent D. Chapman and Ivo Feussner. PMID:27155216

  8. Comprehensive analysis of the major lipid classes in sebum by rapid resolution high-performance liquid chromatography and electrospray mass spectrometry.

    PubMed

    Camera, Emanuela; Ludovici, Matteo; Galante, Marisa; Sinagra, Jo-Linda; Picardo, Mauro

    2010-11-01

    Sebum is a complex lipid mixture that is synthesized in sebaceous glands and excreted on the skin surface. The purpose of this study was the comprehensive detection of the intact lipids that compose sebum. These lipids exist as a broad range of chemical structures and concentrations. Sebum was collected with SebuTape(TM) from the foreheads of healthy donors, and then separated by HPLC on a C8 stationary phase with sub 2 µm particle size. This HPLC method provided high resolution and excellent reproducibility of retention times (RT). Compound mining was performed with time of flight (TOF) and triple quadrupole (QqQ) mass spectrometers (MS), which allowed for the classification of lipids according to their elemental composition, degree of unsaturation, and MS/MS fragmentation. The combination of the two MS systems detected 95 and 29 families of triacylglycerols (TAG) and diacylglycerols (DAG), respectively. Assignment was carried out regardless of positional isomerism. Among the wax esters (WE), 28 species were found to contain the 16:1 fatty acyl moiety. This method was suitable for the simultaneous detection of squalene and its oxygenated derivative. A total of 9 cholesterol esters (CE) were identified and more than 48 free fatty acids (FFA) were detected in normal sebum. The relative abundance of each individual lipid within its own chemical class was determined for 12 healthy donors. In summary, this method provided the first characterization of the features and distribution of intact components of the sebum lipidome. PMID:20719760

  9. Electromagnetic unification of matter and force fields

    NASA Astrophysics Data System (ADS)

    John, Sarah

    2004-05-01

    Special relativity and quantum mechanics are descriptive of electromagnetic propagation in waveguides, with mass analogous to the cutoff frequency of a waveguide mode [S.John, Bull.Am.Phys.Soc. vol.39,no.2,1254 (1994)]. It is further postulated herein that all spin 1/2 matter (necessarily massive) and spin 1 force fields have their origin in the electromagnetic fields E and B. This concept is not new. Majorana, among others have obtained electromagnetic representations of Dirac-like equations valid for the zero-mass case. Here, the spinor representation of the Maxwell equations, as given by Sallhofer, is extended to oscillatory fields with propagation constant m to obtain, in the absence of charge and current densities, the coupled equation (M. hatp + β E)ψ = 0 , where M = diag[ M σ, M^* σ ] , β = offdiag[I,I] , ψ ^ = i ^dag ( σ. B0 ( p), σ. E_0(p)), and M=m+ip, with the energy-mass relation given by E^2 = M M . Further, it is shown that the interaction term of QED is a direct consequence of including the sources and currents of Maxwell equations. Qualitative field patterns for spin 1/2 and spin 1 states, such as the electron, neutrino, magnetic monopole, quarks, photon, and massive gauge bosons are suggested.

  10. Tailoring the Synergistic Bronsted-Lewis acidic effects in Heteropolyacid catalysts: Applied in Esterification and Transesterification Reactions.

    PubMed

    Tao, Meilin; Xue, Lifang; Sun, Zhong; Wang, Shengtian; Wang, Xiaohong; Shi, Junyou

    2015-01-01

    In order to investigate the influences of Lewis metals on acidic properties and catalytic activities, a series of Keggin heteropolyacid (HPA) catalysts, HnPW11MO39 (M = Ti(IV), Cu(II), Al(III), Sn(IV), Fe(III), Cr(III), Zr(IV) and Zn(II); for Ti and Zr, the number of oxygen is 40), were prepared and applied in the esterification and transesterification reactions. Only those cations with moderate Lewis acidity had a higher impact. Ti Substituted HPA, H5PW11TiO40, posse lower acid content compared with Ti(x)H(3-4x)PW12O40 (Ti partial exchanged protons in saturated H3PW12O40), which demonstrated that the Lewis metal as an addenda atom (H5PW11TiO40) was less efficient than those as counter cations (Ti(x)H(3-4x)PW12O40). On the other hand, the highest conversion reached 92.2% in transesterification and 97.4% in esterification. Meanwhile, a good result was achieved by H5PW11TiO40 in which the total selectivity of DAG and TGA was 96.7%. In addition, calcination treatment to H5PW11TiO40 make it insoluble in water which resulted in a heterogeneous catalyst feasible for reuse. PMID:26374393

  11. Characterization of Brown spider (Loxosceles intermedia) hemolymph: cellular and biochemical analyses.

    PubMed

    Bednaski, A V; Trevisan-Silva, D; Matsubara, F H; Boia-Ferreira, M; Olivério, M M; Gremski, L H; Cavalheiro, R P; De Paula, D M B; Paredes-Gamero, E J; Takahashi, H K; Toledo, M S; Nader, H B; Veiga, S S; Chaim, O M; Senff-Ribeiro, A

    2015-05-01

    This is the first study on the hemolymph from a spider of the Loxosceles genus. These animals are responsible for a great number of envenomation cases worldwide. Several studies on Loxosceles venoms have been published, and the knowledge about the venom and its toxins is considerable, not only regarding the biological and biochemical characterization, but also regarding structural, genetic and phylogenetic approaches. However, the literature on Loxosceles hemolymph is nonexistent. The main goal of the present study was to characterize biochemically the hemolymph content, and especially, to identify its different hemocytes. Moreover, many papers have already shown molecules whose source is the hemolymph and their very interesting activities and biomedical applications, for example, antifungal and antibacterial activities. A 2D-SDS-PAGE of brown spider hemolymph showed approximately 111 spots for pH 3-10 and 150 spots for pH 4-7. A lectin-blotting assay showed that hemolymph carbohydrate residues were similar to those found in venom. Several types of TAG and DAG phospholipids were found in the hemolymph and characterized by HPTLC and mass spectrometry. Four different hemocytes were characterized in Loxosceles intermedia hemolymph: prohemocyte, plasmatocyte, granulocyte and adipohemocyte. This paper opens new possibilities on toxinology, studying an unknown biological material, and it characterizes a source of molecules with putative biotechnological applications. PMID:25720299

  12. Diminished Macrophage Apoptosis and Reactive Oxygen Species Generation after Phorbol Ester Stimulation in Crohn's Disease

    PubMed Central

    Palmer, Christine D.; Rahman, Farooq Z.; Sewell, Gavin W.; Ahmed, Afshan; Ashcroft, Margaret; Bloom, Stuart L.; Segal, Anthony W.; Smith, Andrew M.

    2009-01-01

    Background Crohn's Disease (CD) is a chronic relapsing disorder characterized by granulomatous inflammation of the gastrointestinal tract. Although its pathogenesis is complex, we have recently shown that CD patients have a systemic defect in macrophage function, which results in the defective clearance of bacteria from inflammatory sites. Methodology/Principal Findings Here we have identified a number of additional macrophage defects in CD following diacylglycerol (DAG) homolog phorbol-12-myristate-13-acetate (PMA) activation. We provide evidence for decreased DNA fragmentation, reduced mitochondrial membrane depolarization, impaired reactive oxygen species production, diminished cytochrome c release and increased IL-6 production compared to healthy subjects after PMA exposure. The observed macrophage defects in CD were stimulus-specific, as normal responses were observed following p53 activation and endoplasmic reticulum stress. Conclusion These findings add to a growing body of evidence highlighting disordered macrophage function in CD and, given their pivotal role in orchestrating inflammatory responses, defective apoptosis could potentially contribute to the pathogenesis of CD. PMID:19907654

  13. Compression of Probabilistic XML Documents

    NASA Astrophysics Data System (ADS)

    Veldman, Irma; de Keijzer, Ander; van Keulen, Maurice

    Database techniques to store, query and manipulate data that contains uncertainty receives increasing research interest. Such UDBMSs can be classified according to their underlying data model: relational, XML, or RDF. We focus on uncertain XML DBMS with as representative example the Probabilistic XML model (PXML) of [10,9]. The size of a PXML document is obviously a factor in performance. There are PXML-specific techniques to reduce the size, such as a push down mechanism, that produces equivalent but more compact PXML documents. It can only be applied, however, where possibilities are dependent. For normal XML documents there also exist several techniques for compressing a document. Since Probabilistic XML is (a special form of) normal XML, it might benefit from these methods even more. In this paper, we show that existing compression mechanisms can be combined with PXML-specific compression techniques. We also show that best compression rates are obtained with a combination of PXML-specific technique with a rather simple generic DAG-compression technique.

  14. Lipid classes and fatty acid regiodistribution in triacylglycerols of seed oils of two Sambucus species (S. nigra L. and S. ebulus L.).

    PubMed

    Dulf, Francisc Vasile; Oroian, Ioan; Vodnar, Dan Cristian; Socaciu, Carmen; Pintea, Adela

    2013-01-01

    The oil content and fatty acid composition of total lipids (TLs) and main lipid classes (NLs- neutral and PLs- polar lipids) in seeds of two wild Sambucus species (S. nigra and S. ebulus) from Transylvania (Romania) were determined by capillary gas chromatography (GC-MS). In addition, the positional distribution of fatty acids in seed triacylglycerols (TAGs) was determined by hydrolysis with pancreatic lipase. The seeds were found to be rich in fat (22.40-24.90 g/100g) with high amounts of polyunsaturated fatty acids (PUFAs) ranging from 68.96% (S. ebulus) to 75.15% (S. nigra). High ratios of PUFAs/SFAs (saturated fatty acids), ranging from 7.06 (S. nigra) to 7.64 (S. ebulus), and low ratios of n-6/n-3, ranging from 0.84 (S. nigra) to 1.51 (S. ebulus), were determined in both oils. The lipid classes/subclasses analyzed (PLs, MAGs--monoacylglycerols, DAGs--diacylglycerols, FFAs--free fatty acids, TAGs and SEs--sterol esters) were separated and identified using thin-layer chromatography. The fatty acid compositions of the TAG fractions were practically identical to the profiles of TLs, with the same dominating fatty acids in both analyzed species. SEs and FFAs, were characterized by high proportions of SFAs. The sn-2 position of TAGs was esterified predominantly with linoleic acid (43.56% for S. nigra and 50.41% for S. ebulus). PMID:24071984

  15. Signaling pathways in diabetic nephropathy.

    PubMed

    Kawanami, Daiji; Matoba, Keiichiro; Utsunomiya, Kazunori

    2016-10-01

    Diabetic nephropathy (DN) is a major cause of end-stage renal disease (ESRD), however, specific treatment for DN has not yet been elucidated. Therefore, it is critically important to understand the molecular mechanism underlying DN to develop cause-related therapeutic strategy. To date, various factors such as hemodynamic changes and metabolic pathways have been shown to be involved in the pathogenesis of DN. Excessive glucose influx activates cellular signaling pathways, including the diacylglycerol (DAG)-protein kinase C (PKC) pathway, advanced glycation end-products (AGE), polyol pathway, hexosamine pathway and oxidative stress. These factors interact with one another, thereby facilitating inflammatory processes, leading to the development of glomerulosclerosis under diabetic conditions. In addition to metabolic pathways, Rho-kinase, an effector of small-GTPase binding protein Rho, has been implicated as an important factor in the pathogenesis of DN. A number of studies have demonstrated that Rho-kinase plays key roles in the development of DN by inducing endothelial dysfunction, mesangial excessive extracellular matrix (ECM) production, podocyte abnormality, and tubulointerstitial fibrosis. In this review article, we describe our current understanding of the signaling pathways in DN. PMID:27094540

  16. Time Evolution and Characteristic Quantities of Squeezed Chaotic Field in Diffusion Channel

    NASA Astrophysics Data System (ADS)

    Da, Cheng; Fan, Hong-Yi

    2016-06-01

    In exploring the time evolution law of squeezed chaotic state, described by the density operator, ρ 0=(1-ek) S^{dagger }(r) e^{ka^{dagger }a}S(r) , in a diffusion channel, we find two physical quantities characteristic of this physical process, they are τ=1/( 2bar{n+1) e^{-2r}+1}, θ={1}/{( 2bar{n}+1) e^{2r}+1}, where bar {n} is average photon number of the chaotic field, r is the squeezing parameter and ρ 0 in normal ordering is ρ0=2√{τ θ}\\colon exp [ 1/2( τ -θ ) ( a^{dagger2}+a2) -( τ +θ ) a^{dag}a] \\colon. We find in the diffusion process, τ and 𝜃 evolves into τ → τ^'=τ/1+2κ tτ, θ → θ^'=θ/1+2κ tθ, where κ represent diffusion coefficient, thus ρ ( t) =2√{τ^'θ^'\\colon exp [ 1/2( τ^'-θ^') ( a^{dagger 2}+a2) -( τ^'+θ^') a^{dagger }a] \\colon, this is the evolution law of squeezed chaotic state in diffusion channel. The photon number of the final state slightly increases by an amount κ t. This diffusion process can be considered a quantum controlling scheme in the way of photon addition by adjusting κ.

  17. Recent progress on type II diacylglycerol kinases: the physiological functions of diacylglycerol kinase δ, η and κ and their involvement in disease.

    PubMed

    Sakai, Hiromichi; Sakane, Fumio

    2012-11-01

    Diacylglycerol kinase (DGK) phosphorylates diacylglycerol (DAG) to produce phosphatidic acid (PA) and plays an important role in signal transduction by modulating the balance between these signalling lipids. To date, 10 mammalian DGK isozymes have been identified, and these isozymes are subdivided into five groups according to their structural features. The type II DGKs, consisting of δ1, δ2, η1, η2 and κ isoforms, possess a pleckstrin homology (PH) domain at their N-termini in addition to the separate catalytic region. Moreover, DGKs δ1, δ2 and η2 have a sterile α motif domain at their C-termini. Recent studies have revealed that type II DGKs play pivotal roles in a wide variety of mammalian signal transduction pathways for cell proliferation and differentiation and glucose metabolism and that the DGKs are involved in cancer, type II diabetes, seizures, hypospadias and bipolar disorder. This review summarizes the current knowledge on the properties and physiological functions of type II DGKs and their involvement in disease. PMID:22984004

  18. Gene expression profiling gut microbiota in different races of humans

    NASA Astrophysics Data System (ADS)

    Chen, Lei; Zhang, Yu-Hang; Huang, Tao; Cai, Yu-Dong

    2016-03-01

    The gut microbiome is shaped and modified by the polymorphisms of microorganisms in the intestinal tract. Its composition shows strong individual specificity and may play a crucial role in the human digestive system and metabolism. Several factors can affect the composition of the gut microbiome, such as eating habits, living environment, and antibiotic usage. Thus, various races are characterized by different gut microbiome characteristics. In this present study, we studied the gut microbiomes of three different races, including individuals of Asian, European and American races. The gut microbiome and the expression levels of gut microbiome genes were analyzed in these individuals. Advanced feature selection methods (minimum redundancy maximum relevance and incremental feature selection) and four machine-learning algorithms (random forest, nearest neighbor algorithm, sequential minimal optimization, Dagging) were employed to capture key differentially expressed genes. As a result, sequential minimal optimization was found to yield the best performance using the 454 genes, which could effectively distinguish the gut microbiomes of different races. Our analyses of extracted genes support the widely accepted hypotheses that eating habits, living environments and metabolic levels in different races can influence the characteristics of the gut microbiome.

  19. Regulation of Hepatic Triacylglycerol Metabolism by CGI-58 Does Not Require ATGL Co-activation.

    PubMed

    Lord, Caleb C; Ferguson, Daniel; Thomas, Gwynneth; Brown, Amanda L; Schugar, Rebecca C; Burrows, Amy; Gromovsky, Anthony D; Betters, Jenna; Neumann, Chase; Sacks, Jessica; Marshall, Stephanie; Watts, Russell; Schweiger, Martina; Lee, Richard G; Crooke, Rosanne M; Graham, Mark J; Lathia, Justin D; Sakaguchi, Takuya F; Lehner, Richard; Haemmerle, Guenter; Zechner, Rudolf; Brown, J Mark

    2016-07-26

    Adipose triglyceride lipase (ATGL) and comparative gene identification 58 (CGI-58) are critical regulators of triacylglycerol (TAG) turnover. CGI-58 is thought to regulate TAG mobilization by stimulating the enzymatic activity of ATGL. However, it is not known whether this coactivation function of CGI-58 occurs in vivo. Moreover, the phenotype of human CGI-58 mutations suggests ATGL-independent functions. Through direct comparison of mice with single or double deficiency of CGI-58 and ATGL, we show here that CGI-58 knockdown causes hepatic steatosis in both the presence and absence of ATGL. CGI-58 also regulates hepatic diacylglycerol (DAG) and inflammation in an ATGL-independent manner. Interestingly, ATGL deficiency, but not CGI-58 deficiency, results in suppression of the hepatic and adipose de novo lipogenic program. Collectively, these findings show that CGI-58 regulates hepatic neutral lipid storage and inflammation in the genetic absence of ATGL, demonstrating that mechanisms driving TAG lipolysis in hepatocytes differ significantly from those in adipocytes. PMID:27396333

  20. Evaluation of a Terminal Area In-Trail Approach Spacing, Project and Study

    NASA Technical Reports Server (NTRS)

    Shelden, Stephen; Johnson, Walter (Editor)

    2001-01-01

    Reported here are the results of work completed as a precursor to Distributed Air Ground (DAG), Concept Element 11 (CE11) research. CE11 is a NASA, Advanced Air Transportation Technologies (AATT) concept initiated to promote research on in-trail merging and spacing during approach in the terminal area environment, such that improvements to the National Air Space (NAS) may be realized. A description of the project concept, results of a preliminary study, and a literature review are presented. In terms of conclusions, study results, and reference material respectively: 1) the concept is supported as being significant to NAS capacity improvement, 2) the preliminary study indicated that having more than one downstream, in-trail aircraft present on a CDTI during approach may be advantageous, and that flying traditional Standard Terminal Arrival Routes (STAR) utilizing advanced decision support tools (DST) may be complicated by wake turbulence considerations, a lack of vertical awareness on the part of the flight crew, and the 'step-down' nature of many terminal area approaches, and 3) the results of a literature review are presented for future reference.

  1. Tailoring the Synergistic Bronsted-Lewis acidic effects in Heteropolyacid catalysts: Applied in Esterification and Transesterification Reactions

    PubMed Central

    Tao, Meilin; Xue, Lifang; Sun, Zhong; Wang, Shengtian; Wang, Xiaohong; Shi, Junyou

    2015-01-01

    In order to investigate the influences of Lewis metals on acidic properties and catalytic activities, a series of Keggin heteropolyacid (HPA) catalysts, HnPW11MO39 (M = TiIV, CuII, AlIII, SnIV, FeIII, CrIII, ZrIV and ZnII; for Ti and Zr, the number of oxygen is 40), were prepared and applied in the esterification and transesterification reactions. Only those cations with moderate Lewis acidity had a higher impact. Ti Substituted HPA, H5PW11TiO40, posse lower acid content compared with TixH3−4xPW12O40 (Ti partial exchanged protons in saturated H3PW12O40), which demonstrated that the Lewis metal as an addenda atom (H5PW11TiO40) was less efficient than those as counter cations (TixH3−4xPW12O40). On the other hand, the highest conversion reached 92.2% in transesterification and 97.4% in esterification. Meanwhile, a good result was achieved by H5PW11TiO40 in which the total selectivity of DAG and TGA was 96.7%. In addition, calcination treatment to H5PW11TiO40 make it insoluble in water which resulted in a heterogeneous catalyst feasible for reuse. PMID:26374393

  2. The effects of high-fat feeding on physical function and skeletal muscle extracellular matrix.

    PubMed

    Tam, C S; Power, J E; Markovic, T P; Yee, C; Morsch, M; McLennan, S V; Twigg, S M

    2015-01-01

    Skeletal muscle extracellular matrix (ECM) remodelling has been proposed as a feature of the pathogenic milieu associated with obesity and metabolic dysfunction. Whether muscle ECM is associated with impaired physical function in obese conditions is unknown. C57BL/6 mice were fed a high-fat diet (HFD) or chow for 5, 10 and 25 weeks. Non-invasive physiological tests (hang wire, hang mesh and grip strength) to assess neuromuscular function and motor co-ordination were performed. Genes related to ECM structure (COL1, COL3, COL6A2, SPARC), growth factors (TGFB1, TGFB2, CTGF, VEGF) and muscle function (DMD (Dp147), CPN3, DAG1) were measured in gastrocnemius muscle using real-time PCR and COL1, 3 and 6 protein were measured by western immunoblot. Compared with chow, HFD mice had two to six-fold lower muscle strength (hang wire test; raw data and multiplied by body weight) at all time-points (P<0.001) and two-fold lower hang mesh and grip strength at 10 weeks (P<0.05). At 5 weeks, COL1, COL3 and COL6 gene expression, but not protein levels were three to eight-fold lower in HFD compared with chow. In the HFD group at 5 weeks, greater COL3 and 6 gene expression were associated with poorer hang wire performance. For the first time, our results demonstrate links between muscle ECM structure and physical function in obesity. PMID:26657013

  3. Traffic Behavior Recognition Using the Pachinko Allocation Model.

    PubMed

    Huynh-The, Thien; Banos, Oresti; Le, Ba-Vui; Bui, Dinh-Mao; Yoon, Yongik; Lee, Sungyoung

    2015-01-01

    CCTV-based behavior recognition systems have gained considerable attention in recent years in the transportation surveillance domain for identifying unusual patterns, such as traffic jams, accidents, dangerous driving and other abnormal behaviors. In this paper, a novel approach for traffic behavior modeling is presented for video-based road surveillance. The proposed system combines the pachinko allocation model (PAM) and support vector machine (SVM) for a hierarchical representation and identification of traffic behavior. A background subtraction technique using Gaussian mixture models (GMMs) and an object tracking mechanism based on Kalman filters are utilized to firstly construct the object trajectories. Then, the sparse features comprising the locations and directions of the moving objects are modeled by PAMinto traffic topics, namely activities and behaviors. As a key innovation, PAM captures not only the correlation among the activities, but also among the behaviors based on the arbitrary directed acyclic graph (DAG). The SVM classifier is then utilized on top to train and recognize the traffic activity and behavior. The proposed model shows more flexibility and greater expressive power than the commonly-used latent Dirichlet allocation (LDA) approach, leading to a higher recognition accuracy in the behavior classification. PMID:26151213

  4. The Temperature Dependence of Micro-Leakage between Restorative and Pulp Capping Materials by Cu Diffusion.

    PubMed

    H, Kamalak; A, Mumcu; S, Altin

    2015-01-01

    We used the Cu ions for the leakage analysis between pulp capping and restorative materials. Theoretically, Cu has more advantages than Ag ions due to their smaller radii (rCu (2+)=73 pm and rAg (2+)=94 pm), lower mass density (dCu=8.96 g/cm(3) and dAg=10.49 g/cm(3)) and higher radio opacity which can be more useful by X-ray or EDX detectors, cheaper price and more abundance in planet when compared with Ag element which is generally used in the leakage studies. The micro leakage between dental restorations and pulp capping materials has been determined by using Micro Computed Tomography, Scanning Electron Microscopy and EDX analysis. It is found that the leakage has temperature dependent mechanism which increases with the increasing temperature. As a result, using Cu solution for leakage studies in dentine is an effective and easy method which can be used in dental science. PMID:25926897

  5. Generalized Causal Mediation Analysis

    PubMed Central

    Albert, Jeffrey M.; Nelson, Suchitra

    2010-01-01

    Summary The goal of mediation analysis is to assess direct and indirect effects of a treatment or exposure on an outcome. More generally, we may be interested in the context of a causal model as characterized by a directed acyclic graph (DAG), where mediation via a specific path from exposure to outcome may involve an arbitrary number of links (or ‘stages’). Methods for estimating mediation (or pathway) effects are available for a continuous outcome and a continuous mediator related via a linear model, while for a categorical outcome or categorical mediator, methods are usually limited to two-stage mediation. We present a method applicable to multiple stages of mediation and mixed variable types using generalized linear models. We define pathway effects using a potential outcomes framework and present a general formula that provides the effect of exposure through any specified pathway. Some pathway effects are nonidentifiable and their estimation requires an assumption regarding the correlation between counterfactuals. We provide a sensitivity analysis to assess of the impact of this assumption. Confidence intervals for pathway effect estimates are obtained via a bootstrap method. The method is applied to a cohort study of dental caries in very low birth weight adolescents. A simulation study demonstrates low bias of pathway effect estimators and close-to-nominal coverage rates of confidence intervals. We also find low sensitivity to the counterfactual correlation in most scenarios. PMID:21306353

  6. Voyages of the Zodiac, an Impenitent Traveller across Lands and Ages

    NASA Astrophysics Data System (ADS)

    Belmonte, Juan Antonio

    2015-05-01

    The zodiac is one of humankind's oldest astronomical heritages. Indeed, it has a long history and had an impressive cultural influence in the past that could be traced from the steppes of Mesopotamia to the wilderness of the Roman frontiers in Great Britain. In the present essay, we will discuss the origin of the zodiacal constellations in ancient Mesopotamia from their possible prehistoric ancestors in the pre-ceramic context of the 10,000-year-old site of Göbleki Tepe. Later on, we will discuss the role that the zodiac played in the development of the new cult established by King Antiochos I of Commagene in his hierothesion at Nemrud Dag, where a planetary conjunction in Leo has played a role in the clues to understand the enigmas of such an impressive monument. This will be followed by the analysis of its southwards travel to the Valley of the Nile, where we will study the famous Zodiac of Dendera. Then we will travel westwards to discuss the astronomical aspects of a new religion where astronomical eras and the zodiac ought to play a most relevant role, Mithraism. Finally, new ideas for future research in this most interesting topic will briefly be sketched.

  7. Desnutrin/ATGL is Regulated by AMPK and is Required for a Brown Adipose Phenotype

    PubMed Central

    Ahmadian, Maryam; Abbott, Marcia J.; Tang, Tianyi; Hudak, Carolyn S.S.; Kim, Yangha; Bruss, Matthew; Hellerstein, Marc K.; Lee, Hui-Young; Samuel, Varman T.; Shulman, Gerald I.; Wang, Yuhui; Duncan, Robin E.; Kang, Chulho; Sul, Hei Sook

    2011-01-01

    SUMMARY While fatty acids (FAs) released by white adipose tissue (WAT) provide energy for other organs, lipolysis is also critical in brown adipose tissue (BAT), generating FAs for oxidation and UCP-1 activation for thermogenesis. Here, we show that adipose-specific ablation of desnutrin/ATGL in mice converts BAT to a WAT-like tissue. These mice exhibit severely impaired thermogenesis with increased expression of WAT-enriched genes but decreased BAT genes including UCP-1 with lower PPARα binding to its promoter, revealing the requirement of desnutrin-catalyzed lipolysis for maintaining BAT phenotype. We also show that desnutrin is phosphorylated by AMPK at S406, increasing TAG hydrolase activity, and provide evidence for increased lipolysis by AMPK phosphorylation of desnutrin in adipocytes and in vivo. Despite adiposity and impaired BAT function, desnutrin-ASKO mice have improved hepatic insulin sensitivity with lower DAG levels. Overall, desnutrin is phosphorylated/activated by AMPK to increase lipolysis and brings FA oxidation and UCP-1 induction for thermogenesis. PMID:21641555

  8. The Simulacrum System as a Construct for Mass Spectrometry of Triacylglycerols and Others.

    PubMed

    Byrdwell, William Craig

    2016-02-01

    A construct called a simulacrum is defined that provides all possible solutions to a sum of two mass spectral abundances, based on values (abundances) or ratios of those values. The defined construct is applied to atmospheric pressure chemical ionization mass spectrometry (MS) of triacylglycerols (TAG). A simulacrum has precisely defined components, specifically a simulacrum sum, four Possibilities to Observe, two Cases, and eight solutions. A simulacrum with no restrictions is the First General Form of a Simulacrum. When one value is specified to be 1 (as in MS), the construct is called a Unit Simulacrum, also called the First Specified Form of a Simulacrum. When one value is 1 and no value can be greater than 1 (the two specifications dictated by mass spectrometry), the construct is called the Second Specified Form of a Simulacrum, or the Mass Spectrometry Simulacrum. Simulacra are used with three Critical Ratios calculated from raw abundances in mass spectra of TAG to provide structural information about the degree of unsaturation in TAG, the identity and quantity of regioisomers, and other structural characteristics. Three-level-deep nested simulacrum solutions yield the recently reported Updated Bottom Up Solution, from which the protonated molecule, [MH](+), and all diacylglycerol-like fragments, [DAG](+), of TAG can be reproduced from the Critical Ratios. Thus, the simulacrum solutions constitute a reduced data set in which more information is provided in fewer values than raw abundances, such that the Critical Ratios constitute a compact library of mass spectra. PMID:26662048

  9. Are G-protein-coupled receptors involved in mediating larval settlement and metamorphosis of coral planulae?

    PubMed

    Tran, Cawa; Hadfield, Michael G

    2012-04-01

    Larvae of the scleractinian coral Pocillopora damicornis are induced to settle and metamorphose by the presence of marine bacterial biofilms, and the larvae of Montipora capitata respond to a combination of filamentous and crustose coralline algae. The primary goal of this study was to better understand metamorphosis of cnidarian larvae by determining what types of receptors and signal-transduction pathways are involved during stimulation of metamorphosis of P. damicornis and M. capitata. Evidence from studies on larvae of hydrozoans suggests that G-protein-coupled receptors (GPCRs) are good candidates. Settlement experiments were conducted in which competent larvae were exposed to neuropharmacological agents that affect GPCRs and their associated signal-transduction pathways, AC/cAMP and PI/DAG/PKC. On the basis of the results of these experiments, we conclude that GPCRs and these pathways do not mediate settlement and metamorphosis in either coral species. Two compounds that had an effect on both species, forskolin and phorbol-12-myristate-13-acetate (TPA), may be acting on other cellular processes not related to GPCRs. This study strengthens our understanding of the underlying physiological mechanisms that regulate metamorphosis in coral larvae. PMID:22589403

  10. Lipase-catalyzed esterification of 2-monoricinolein for 1,2 (2,3)-diricinolein synthesis.

    PubMed

    Turner, Charlotta; Wani, Seiji; Wong, Rosalind; Lin, Jiann-Tsyh; McKeon, Thomas

    2006-01-01

    The purpose of this investigation was to develop conditions for producing 2-monoricinoleoyl DAG. We used lipase-catalyzed hydrolysis of triricinolein to obtain 2-monoricinolein and thereafter synthesized 1,2(2,3)-diricinolein through esterification of 2-monoricinolein, using ricinoleic acid as the acyl donor. Five different 1,3-specific immobilized lipases were tested for the initial methanolysis reaction: Candida antarctica type B, Rhizomucor miehei, Rhizopus oryzae (ROL), Thermomyces lanuginosus, and Aspergillus niger. For the second esterification reaction, we investigated these five lipases plus Pseudomonas cepacia, Penicillium roquefortii, Candida rugosa, and Pseudomonas fluorescence. Toluene and diisopropyl ether (DIPE) were examined as reaction media at a water activity of 0.11. ROL in DIPE gave the highest yield of 2-monoricinolein from triricinolein, 78% after 3 h of reaction. The isolated 2-monoricinolein was esterified with ricinoleic acid for synthesis of 1,2(2,3)-diricinolein. ROL in DIPE gave the highest yield of 1,2(2,3)-diricinolein, 58% after 1 h of reaction, and NMR analysis showed that the purity was 97.2%. This methodology can be used for synthesizing radiolabeled 1,2(2,3)-diricinolein to study lipid biosynthesis in castor and other oilseeds. PMID:16555475

  11. Phospholipid: diacylglycerol acyltransferase contributes to the conversion of membrane lipids into triacylglycerol in Myrmecia incisa during the nitrogen starvation stress

    PubMed Central

    Liu, Xiao-Yu; Ouyang, Long-Ling; Zhou, Zhi-Gang

    2016-01-01

    In addition to the Kennedy pathway for de novo biosynthesis, triacylglycerol (TAG), the most important stock for microalgae-based biodiesel production, can be synthesized by phospholipid: diacylglycerol acyltransferase (PDAT) that transfers an acyl group from phospholipids (PLs) to diacylglycerol (DAG). This study presents a novel gene that encodes PDAT from the green microalga Myrmecia incisa Reisigl H4301 (designated MiPDAT ). MiPDAT is localized on the plasma membrane (PM) via the agroinfiltration of tobacco leaves with a green fluorescent protein-fused construct. MiPDAT synthesizes TAG based on functional complementary experiments in the mutant yeast strain H1246 and the membrane lipid phosphatidylcholine (PC) is preferentially used as substrates as revealed by in vitro enzyme activity assay. The gradually increased transcription levels of MiPDAT in M. incisa during the cultivation under nitrogen starvation conditions is proposed to be responsible for the decrease and increase of the PC and TAG levels, respectively, as detected by liquid chromatography-mass spectrometry after 4 d of nitrogen starvation. In addition, the mechanism by which MiPDAT in this microalga uses PC to yield TAG is discussed. Accordingly, it is concluded that this PM-located PDAT contributes to the conversion of membrane lipids into TAG in M. incisa during the nitrogen starvation stress. PMID:27216435

  12. Phosphatidate phosphatase-1 is functionally conserved in lipid synthesis and storage from human to yeast.

    PubMed

    Fang, Zhijia; Wang, Song; Du, Xiuxiu; Shi, Ping; Huang, Zhiwei

    2014-12-01

    Phosphatidate phosphatase-1 (PAP1) enzymes (yeast Pah1p/Smp2p, mammalian lipin1-3) have a key role in lipid homeostasis by controlling the relative proportions of its substrate phosphatidate (PA) and its product diacylglycerol (DAG). Recent investigation shows that mammalian lipin-1 complements phenotypes exhibited by yeast pah1Δ mutant cells, which indicates the functions of PAP1 enzymes are evolutionarily conserved. The observation was confirmed after transformation of human LPIN1 into PAH1-defective yeast, which resulted in human LPIN1-induced accumulation of triacylglycerol (TAG )and lipid droplet formation. In double mutants lacking Tgl3p and Tgl4p, overexpression of PAH1 or LPIN1 induced TAG accumulation and excessive obesity. Furthermore, the obese yeast was used as a model to study the anti-obesity effects of PAP1 activity inhibitors, including propranolol and clenbuterol. The data showed that the inhibitors significantly suppressed TAG accumulation and lipid droplets formation. These findings demonstrate that LPIN1 plays a functional role in lipid synthesis and storage, a role which is highly conserved from human to yeast. Inhibition of TAG synthesis will become an efficacious treatment strategy for obesity and our excessive obesity model will provide a very useful tool for discovery of new anti-obesity drugs in the future. PMID:25475986

  13. Conditional control of gene expression by synthetic riboswitches in Streptomyces coelicolor.

    PubMed

    Rudolph, Martin M; Vockenhuber, Michael-Paul; Suess, Beatrix

    2015-01-01

    Here we provide a step-by-step protocol for the application of synthetic theophylline-dependent riboswitches for conditional gene expression in Streptomyces coelicolor. Application of the method requires a sequence of only ~85 nt to be inserted between the transcriptional start site and the start codon of a gene of interest. No auxiliary factors are needed. All tested riboswitch variants worked well in concert with the promoters galP2, ermEp1, and SF14. Moreover, they allowed theophylline-dependent expression not only of the heterologous β-glucuronidase reporter gene but also of dagA, an endogenous agarase gene. The right combination of the tested promoters with the riboswitch variants allows for the adjustment of the desired dynamic range of regulation in a highly specific and dose-dependent manner and underlines the orthogonality of riboswitch regulation. We anticipate that any additional natural or synthetic promoter can be combined with the presented riboswitches. Moreover, this system should easily be transferable to other Streptomyces species, and most likely to any other genetically manipulable bacteria. PMID:25605391

  14. Insulin Resistance and Obesity Affect Lipid Profile in the Salivary Glands.

    PubMed

    Matczuk, Jan; Zalewska, Anna; Łukaszuk, Bartłomiej; Knaś, Małgorzata; Maciejczyk, Mateusz; Garbowska, Marta; Ziembicka, Dominika M; Waszkiel, Danuta; Chabowski, Adrian; Żendzian-Piotrowska, Małgorzata; Kurek, Krzysztof

    2016-01-01

    In today's world wrong nutritional habits together with a low level of physical activity have given rise to the development of obesity and its comorbidity, insulin resistance. More specifically, many researches indicate that lipids are vitally involved in the onset of a peripheral tissue (e.g., skeletal muscle, heart, and liver) insulin resistance. Moreover, it seems that diabetes can also induce changes in respect of lipid composition of both the salivary glands and saliva. However, judging by the number of research articles, the salivary glands lipid profile still has not been sufficiently explored. In the current study we aim to assess the changes in the main lipid fractions, namely, triacylglycerols, phospholipids, free fatty acids, and diacylglycerols, in the parotid and the submandibular salivary glands of rats exposed to a 5-week high fat diet regimen. We observed that the high caloric fat diet caused a significant change in the salivary glands lipid composition, especially with respect to PH and TG, but not DAG or FFAs, classes. The observed reduction in PH concentration is an interesting phenomenon frequently signifying the atrophy and malfunctions in the saliva secreting organs. On the other hand, the increased accumulation of TG in the glands may be an important clinical manifestation of metabolic syndrome and type 2 diabetes mellitus. PMID:27471733

  15. Phosphatidic acid phosphatase and diacylglycerol acyltransferase: potential targets for metabolic engineering of microorganism oil.

    PubMed

    Jin, Hong-Hao; Jiang, Jian-Guo

    2015-04-01

    Oleaginous microorganism is becoming one of the most promising oil feedstocks for biodiesel production due to its great advantages in triglyceride (TAG) accumulation. Previous studies have shown that de novo TAG biosynthesis can be divided into two parts: the fatty acid biosynthesis pathway (the upstream part which generates acyl-CoAs) and the glycerol-3-phosphate acylation pathway (the downstream part in which three acyl groups are sequentially added onto a glycerol backbone). This review mainly focuses on two enzymes in the G3P pathway, phosphatidic acid phosphatase (PAP) and diacylglycerol acyltransferase (DGAT). The former catalyzes a dephosphorylation reaction, and the latter catalyzes a subsequent acylation reaction. Genes, functional motifs, transmembrane domains, action mechanism, and new studies of the two enzymes are discussed in detail. Furthermore, this review also covers diacylglycerol kinase, an enzyme that catalyzes the reverse reaction of diacylglycerol formation. In addition, PAP and DGAT are the conjunction points of the G3P pathway, the Kennedy pathway, and the CDP-diacylglycerol pathway (CDP-DAG pathway), and the mutual transformation between TAGs and phospholipids is discussed as well. Given that both the Kennedy and CDP-diacylglycerol pathways are in metabolic interlock (MI) with the G3P pathway, it is suggested that, via metabolic engineering, TAG accumulation can be improved by the two pathways based on the pivotal function of PAP and DGAT. PMID:25672855

  16. The IceProd (IceCube Production) Framework

    NASA Astrophysics Data System (ADS)

    Díaz-Vélez, J. C.

    2014-06-01

    IceProd is a data processing and management framework developed by the IceCube Neutrino Observatory for processing of Monte Carlo simulations and data. IceProd runs as a separate layer on top of middleware or cluster job schedulers and can take advantage of a variety of computing resources including grids such as EGI, OSG, and NorduGrid as well as local clusters running batch systems like HT Condor, PBS, and SGE. This is accomplished by a set of dedicated daemons which process job submission in a coordinated fashion through the use of middleware plug-ins that serve to abstract the details of job submission and job management. IceProd can also manage complex workflow DAGs across distributed computing grids in order to optimize usage of resources. We describe several aspects of IceProd's design and it's applications in collaborative computing environments. We also briefly discuss design aspects of a second generation IceProd, currently being tested in IceCube.

  17. Degree of Cajal-Retzius Cell Mislocalization Correlates with the Severity of Structural Brain Defects in Mouse Models of Dystroglycanopathy.

    PubMed

    Booler, Helen