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Sample records for peach small gtpases

  1. Insights into the classification of small GTPases

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    Dominik Heider

    2010-05-01

    Full Text Available Dominik Heider1, Sascha Hauke3, Martin Pyka4, Daniel Kessler21Department of Bioinformatics, Center for Medical Biotechnology, 2Institute of Cell Biology (Cancer Research, University of Duisburg-Essen, Essen, Germany; 3Institute of Computer Science, University of Münster, Münster, Germany; 4Interdisciplinary Center for Clinical Research, University Hospital of Münster, Münster, GermanyAbstract: In this study we used a Random Forest-based approach for an assignment of small guanosine triphosphate proteins (GTPases to specific subgroups. Small GTPases represent an important functional group of proteins that serve as molecular switches in a wide range of fundamental cellular processes, including intracellular transport, movement and signaling events. These proteins have further gained a special emphasis in cancer research, because within the last decades a huge variety of small GTPases from different subgroups could be related to the development of all types of tumors. Using a random forest approach, we were able to identify the most important amino acid positions for the classification process within the small GTPases superfamily and its subgroups. These positions are in line with the results of earlier studies and have been shown to be the essential elements for the different functionalities of the GTPase families. Furthermore, we provide an accurate and reliable software tool (GTPasePred to identify potential novel GTPases and demonstrate its application to genome sequences.Keywords: cancer, machine learning, classification, Random Forests, proteins

  2. Targeting and localized signalling by small GTPases

    NARCIS (Netherlands)

    ten Klooster, Jean Paul; Hordijk, Peter L.

    2007-01-01

    Polarized cellular responses, for example, cell migration, require the co-ordinated assembly of signalling complexes at a particular subcellular location, such as the leading edge of cells. Small GTPases of the Ras superfamily play central roles in many (polarized) responses to growth factors,

  3. Small GTPases and formins in mammalian oocyte maturation: cytoskeletal organizers.

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    Kwon, Sojung; Lim, Hyunjung J

    2011-03-01

    The maturation process of mammalian oocytes accompanies an extensive rearrangement of the cytoskeleton and associated proteins. As this process requires a delicate interplay between the cytoskeleton and its regulators, it is often targeted by various external and internal adversaries that affect the congression and/or segregation of chromosomes. Asymmetric cell division in oocytes also requires specific regulators of the cytoskeleton, including formin-2 and small GTPases. Recent literature providing clues regarding how actin filaments and microtubules interact during spindle migration in mouse oocytes are highlighted in this review.

  4. Evolution and Diversity of the Ras Superfamily of Small GTPases in Prokaryotes

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    Wuichet, Kristin; Søgaard-Andersen, Lotte

    2015-01-01

    The Ras superfamily of small GTPases are single domain nucleotide-dependent molecular switches that act as highly tuned regulators of complex signal transduction pathways. Originally identified in eukaryotes for their roles in fundamental cellular processes including proliferation, motility, polarity, nuclear transport, and vesicle transport, recent studies have revealed that single domain GTPases also control complex functions such as cell polarity, motility, predation, development and antibiotic resistance in bacteria. Here, we used a computational genomics approach to understand the abundance, diversity, and evolution of small GTPases in prokaryotes. We collected 520 small GTPase sequences present in 17% of 1,611 prokaryotic genomes analyzed that cover diverse lineages. We identified two discrete families of small GTPases in prokaryotes that show evidence of three distinct catalytic mechanisms. The MglA family includes MglA homologs, which are typically associated with the MglB GTPase activating protein, whereas members of the Rup (Ras superfamily GTPase of unknown function in prokaryotes) family are not predicted to interact with MglB homologs. System classification and genome context analyses support the involvement of small GTPases in diverse prokaryotic signal transduction pathways including two component systems, laying the foundation for future experimental characterization of these proteins. Phylogenetic analysis of prokaryotic and eukaryotic GTPases supports that the last universal common ancestor contained ancestral MglA and Rup family members. We propose that the MglA family was lost from the ancestral eukaryote and that the Ras superfamily members in extant eukaryotes are the result of vertical and horizontal gene transfer events of ancestral Rup GTPases. PMID:25480683

  5. Development and application of a quantitative multiplexed small GTPase activity assay using targeted proteomics.

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    Zhang, Cheng-Cheng; Li, Ru; Jiang, Honghui; Lin, Shujun; Rogalski, Jason C; Liu, Kate; Kast, Juergen

    2015-02-06

    Small GTPases are a family of key signaling molecules that are ubiquitously expressed in various types of cells. Their activity is often analyzed by western blot, which is limited by its multiplexing capability, the quality of isoform-specific antibodies, and the accuracy of quantification. To overcome these issues, a quantitative multiplexed small GTPase activity assay has been developed. Using four different binding domains, this assay allows the binding of up to 12 active small GTPase isoforms simultaneously in a single experiment. To accurately quantify the closely related small GTPase isoforms, a targeted proteomic approach, i.e., selected/multiple reaction monitoring, was developed, and its functionality and reproducibility were validated. This assay was successfully applied to human platelets and revealed time-resolved coactivation of multiple small GTPase isoforms in response to agonists and differential activation of these isoforms in response to inhibitor treatment. This widely applicable approach can be used for signaling pathway studies and inhibitor screening in many cellular systems.

  6. Comparative phylogenetic and expression analysis of small GTPases families in legume and non-legume plants.

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    Flores, Ana Claudia; Via, Virginia Dalla; Savy, Virginia; Villagra, Ulises Mancini; Zanetti, María Eugenia; Blanco, Flavio

    2018-02-01

    Small monomeric GTPases act as molecular switches in several processes that involve polar cell growth, participating mainly in vesicle trafficking and cytoskeleton rearrangements. This gene superfamily has largely expanded in plants through evolution as compared with other Kingdoms, leading to the suggestion that members of each subfamily might have acquired new functions associated to plant-specific processes. Legume plants engage in a nitrogen-fixing symbiotic interaction with rhizobia in a process that involves polar growth processes associated with the infection throughout the root hair. To get insight into the evolution of small GTPases associated with this process, we use a comparative genomic approach to establish differences in the Ras GTPase superfamily between legume and non-legume plants. Phylogenetic analyses did not show clear differences in the organization of the different subfamilies of small GTPases between plants that engage or not in nodule symbiosis. Protein alignments revealed a strong conservation at the sequence level of small GTPases previously linked to nodulation by functional genetics. Interestingly, one Rab and three Rop proteins showed conserved amino acid substitutions in legumes, but these changes do not alter the predicted conformational structure of these proteins. Although the steady-state levels of most small GTPases do not change in response to rhizobia, we identified a subset of Rab, Rop and Arf genes whose transcript levels are modulated during the symbiotic interaction, including their spatial distribution along the indeterminate nodule. This study provides a comprehensive study of the small GTPase superfamily in several plant species. The genetic program associated to root nodule symbiosis includes small GTPases to fulfill specific functions during infection and formation of the symbiosomes. These GTPases seems to have been recruited from members that were already present in common ancestors with plants as distant as monocots

  7. The Ins and Outs of Small GTPase Rac1 in the Vasculature

    NARCIS (Netherlands)

    Marinković, Goran; Heemskerk, Niels; van Buul, Jaap D.; de Waard, Vivian

    2015-01-01

    The Rho family of small GTPases forms a 20-member family within the Ras superfamily of GTP-dependent enzymes that are activated by a variety of extracellular signals. The most well known Rho family members are RhoA (Ras homolog gene family, member A), Cdc42 (cell division control protein 42), and

  8. Small GTPases and Stress Responses of vvran1 in the Straw Mushroom Volvariella volvacea

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    Jun-Jie Yan

    2016-09-01

    Full Text Available Small GTPases play important roles in the growth, development and environmental responses of eukaryotes. Based on the genomic sequence of the straw mushroom Volvariella volvacea, 44 small GTPases were identified. A clustering analysis using human small GTPases as the references revealed that V. volvacea small GTPases can be grouped into five families: nine are in the Ras family, 10 are in the Rho family, 15 are in the Rab family, one is in the Ran family and nine are in the Arf family. The transcription of vvran1 was up-regulated upon hydrogen peroxide (H2O2 stress, and could be repressed by diphenyleneiodonium chloride (DPI, a NADPH oxidase-specific inhibitor. The number of vvran1 transcripts also increased upon cold stress. Diphenyleneiodonium chloride, but not the superoxide dismutase (SOD inhibitor diethy dithiocarbamate (DDC, could suppress the up-regulation of vvran1 gene expression to cold stress. These results combined with the high correlations between gene expression and superoxide anion (O2− generation indicated that vvran1 could be one of the candidate genes in the downstream of O2− mediated pathways that are generated by NADPH oxidase under low temperature and oxidative stresses.

  9. Topological and functional properties of the small GTPases protein interaction network.

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    Anna Delprato

    Full Text Available Small GTP binding proteins of the Ras superfamily (Ras, Rho, Rab, Arf, and Ran regulate key cellular processes such as signal transduction, cell proliferation, cell motility, and vesicle transport. A great deal of experimental evidence supports the existence of signaling cascades and feedback loops within and among the small GTPase subfamilies suggesting that these proteins function in a coordinated and cooperative manner. The interplay occurs largely through association with bi-partite regulatory and effector proteins but can also occur through the active form of the small GTPases themselves. In order to understand the connectivity of the small GTPases signaling routes, a systems-level approach that analyzes data describing direct and indirect interactions was used to construct the small GTPases protein interaction network. The data were curated from the Search Tool for the Retrieval of Interacting Genes (STRING database and include only experimentally validated interactions. The network method enables the conceptualization of the overall structure as well as the underlying organization of the protein-protein interactions. The interaction network described here is comprised of 778 nodes and 1943 edges and has a scale-free topology. Rac1, Cdc42, RhoA, and HRas are identified as the hubs. Ten sub-network motifs are also identified in this study with themes in apoptosis, cell growth/proliferation, vesicle traffic, cell adhesion/junction dynamics, the nicotinamide adenine dinucleotide phosphate (NADPH oxidase response, transcription regulation, receptor-mediated endocytosis, gene silencing, and growth factor signaling. Bottleneck proteins that bridge signaling paths and proteins that overlap in multiple small GTPase networks are described along with the functional annotation of all proteins in the network.

  10. Recycling domains in plant cell morphogenesis: small GTPase effectors, plasma membrane signalling and the exocyst.

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    Zárský, Viktor; Potocký, Martin

    2010-04-01

    The Rho/Rop small GTPase regulatory module is central for initiating exocytotically ACDs (active cortical domains) in plant cell cortex, and a growing array of Rop regulators and effectors are being discovered in plants. Structural membrane phospholipids are important constituents of cells as well as signals, and phospholipid-modifying enzymes are well known effectors of small GTPases. We have shown that PLDs (phospholipases D) and their product, PA (phosphatidic acid), belong to the regulators of the secretory pathway in plants. We have also shown that specific NOXs (NADPH oxidases) producing ROS (reactive oxygen species) are involved in cell growth as exemplified by pollen tubes and root hairs. Most plant cells exhibit several distinct plasma membrane domains (ACDs), established and maintained by endocytosis/exocytosis-driven membrane protein recycling. We proposed recently the concept of a 'recycling domain' (RD), uniting the ACD and the connected endosomal recycling compartment (endosome), as a dynamic spatiotemporal entity. We have described a putative GTPase-effector complex exocyst involved in exocytic vesicle tethering in plants. Owing to the multiplicity of its Exo70 subunits, this complex, along with many RabA GTPases (putative recycling endosome organizers), may belong to core regulators of RD organization in plants.

  11. Small GTPases are involved in sprout formation in human granulosa lutein cells.

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    Franz, Maximilian B; Daube, Stefanie; Keck, Christoph; Sator, Michael; Pietrowski, Detlef

    2013-04-01

    The corpus luteum (CL), develops from the ruptured follicle after gonadotropin stimulation. Based on intracellular reorganization of the cytoskeleton an human chorionic gonadotropin (hCG) dependent sprouting and migration of luteinizing granulosa cells (LGCs) and endothelial cells is observed. Rho-GTPases are shown to be key regulators of cytoskeletal restructuring. In the present study we analyzed the role of Rho-GTPases in the sprouting activity of LGCs. We used the Rho-GTPase-inhibitors Toxin A and -B and the Cdc42-activator Bradykinin in a LGC-spheroid sprouting assay to determine the effect of these modulators in LGCs. Toxin A and Toxin B reduces sprout formation in LGC spheroids. However, the reduction is less than in hCG treated cells. The usage of Bradykinin demonstrates both, a reduction of sprouts in untreated spheroids and an increase of sprouting in previous hCG treated spheroids. The presented results let us suggest that small Rho-GTPases may regulate the sprouting activity of LGCs after stimulation by hCG and that this mechanism may play a role in CL formation.

  12. A photocleavable rapamycin conjugate for spatiotemporal control of small GTPase activity.

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    Umeda, Nobuhiro; Ueno, Tasuku; Pohlmeyer, Christopher; Nagano, Tetsuo; Inoue, Takanari

    2011-01-12

    We developed a novel method to spatiotemporally control the activity of signaling molecules. A newly synthesized photocaged rapamycin derivative induced rapid dimerization of FKBP (FK-506 binding protein) and FRB (FKBP-rapamycin binding protein) upon UV irradiation. With this system and the spatially confined UV irradiation, we achieved subcellularly localized activation of Rac, a member of small GTPases. Our technique offers a powerful approach to studies of dynamic intracellular signaling events.

  13. The glyceraldehyde-3-phosphate dehydrogenase and the small GTPase Rab 2 are crucial for Brucella replication.

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    Emilie Fugier

    2009-06-01

    Full Text Available The intracellular pathogen Brucella abortus survives and replicates inside host cells within an endoplasmic reticulum (ER-derived replicative organelle named the "Brucella-containing vacuole" (BCV. Here, we developed a subcellular fractionation method to isolate BCVs and characterize for the first time the protein composition of its replicative niche. After identification of BCV membrane proteins by 2 dimensional (2D gel electrophoresis and mass spectrometry, we focused on two eukaryotic proteins: the glyceraldehyde-3-phosphate dehydrogenase (GAPDH and the small GTPase Rab 2 recruited to the vacuolar membrane of Brucella. These proteins were previously described to localize on vesicular and tubular clusters (VTC and to regulate the VTC membrane traffic between the endoplasmic reticulum (ER and the Golgi. Inhibition of either GAPDH or Rab 2 expression by small interfering RNA strongly inhibited B. abortus replication. Consistent with this result, inhibition of other partners of GAPDH and Rab 2, such as COPI and PKC iota, reduced B. abortus replication. Furthermore, blockage of Rab 2 GTPase in a GDP-locked form also inhibited B. abortus replication. Bacteria did not fuse with the ER and instead remained in lysosomal-associated membrane vacuoles. These results reveal an essential role for GAPDH and the small GTPase Rab 2 in B. abortus virulence within host cells.

  14. Maturation and integration of adult born hippocampal neurons: signal convergence onto small Rho GTPases

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    Krishna eVadodaria

    2013-08-01

    Full Text Available Adult neurogenesis, restricted to specific regions in the mammalian brain, represents one of the most interesting forms of plasticity in the mature nervous system. Adult-born hippocampal neurons play important roles in certain forms of learning and memory, and altered hippocampal neurogenesis has been associated with a number of neuropsychiatric diseases such as major depression and epilepsy. Newborn neurons go through distinct developmental steps from a dividing neurogenic precursor to a synaptically integrated mature neuron. Previous studies have uncovered several molecular signaling pathways involved in distinct steps of this maturational process. In this context, the small Rho GTPases, Cdc42, Rac1 and RhoA have recently been shown to regulate the morphological and synaptic maturation of adult-born dentate granule cells in vivo. Distinct upstream regulators, including several growth factors that modulate maturation and integration of newborn neurons have been shown to also recruit the small Rho GTPases. Here we review recent findings and highlight the possibility that small Rho GTPases may act as central assimilators, downstream of critical input onto adult-born hippocampal neurons contributing to their maturation and integration into the existing dentate gyrus circuitry.

  15. Ablation of p120-Catenin Altering the Activity of Small GTPase in Human Lung Cancer Cells

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    Nan LIU

    2009-05-01

    Full Text Available Background and objective p120-catenin (p120ctn, a member of the Armadillo gene family, has emerged as an important modulator of small GTPase activities. Therefore, it plays novel roles in tumor malignant phenotype, such as invasion and metastasis, whose mechanism are not well clarified yet. The aim of this study is to explore the roles of p120ctn on the regulation of small GTP family members in lung cancer and the effects to lung cancer invasions andmetastasis. Methods After p120ctn was knocked down by siRNA, in vivo and in vitro analysis was applied to investigate the role and possible mechanism of p120ctn in lung cancer, such as Western Blot, pull-down analysis, and nude mice models. Results p120ctn depletion inactivated RhoA, with the the activity of Cdc42 and Rac1 increased, the invasiveness of lung cancer cells was promoted both in vitro and in vivo . Conclusion p120ctn gene knockdown enhances the metastasis of lung cancer cells, probably by altering expression of small GTPase, such as inactivation of RhoA and activation of Cdc42/Rac1.

  16. Rho GTPases and cancer

    DEFF Research Database (Denmark)

    Li, Hui; Peyrollier, Karine; Kilic, Gülcan

    2014-01-01

    Rho GTPases are a family of small GTPases, which play an important role in the regulation of the actin cytoskeleton. Not surprisingly, Rho GTPases are crucial for cell migration and therefore highly important for cancer cell invasion and the formation of metastases. In addition, Rho GTPases...... are involved in growth and survival of tumor cells, in the interaction of tumor cells with their environment, and they are vital for the cancer supporting functions of the tumor stroma. Recent research has significantly improved our understanding of the regulation of Rho GTPase activity, the specificity of Rho...

  17. Essential role of the small GTPase Ran in postnatal pancreatic islet development.

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    Fang Xia

    Full Text Available The small GTPase Ran orchestrates pleiotropic cellular responses of nucleo-cytoplasmic shuttling, mitosis and subcellular trafficking, but whether deregulation of these pathways contributes to disease pathogenesis has remained elusive. Here, we generated transgenic mice expressing wild type (WT Ran, loss-of-function Ran T24N mutant or constitutively active Ran G19V mutant in pancreatic islet β cells under the control of the rat insulin promoter. Embryonic pancreas and islet development, including emergence of insulin(+ β cells, was indistinguishable in control or transgenic mice. However, by one month after birth, transgenic mice expressing any of the three Ran variants exhibited overt diabetes, with hyperglycemia, reduced insulin production, and nearly complete loss of islet number and islet mass, in vivo. Deregulated Ran signaling in transgenic mice, adenoviral over-expression of WT or mutant Ran in isolated islets, or short hairpin RNA (shRNA silencing of endogenous Ran in model insulinoma INS-1 cells, all resulted in decreased expression of the pancreatic and duodenal homeobox transcription factor, PDX-1, and reduced β cell proliferation, in vivo. These data demonstrate that a finely-tuned balance of Ran GTPase signaling is essential for postnatal pancreatic islet development and glucose homeostasis, in vivo.

  18. The Drosophila small GTPase Rac2 is required for normal feeding and mating behaviour.

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    Goergen, Philip; Kasagiannis, Anna; Schiöth, Helgi B; Williams, Michael J

    2014-03-01

    All multicellular organisms require the ability to regulate bodily processes in order to maintain a stable condition, which necessitates fluctuations in internal metabolics, as well as modifications of outward behaviour. Understanding the genetics behind this modulation is important as a general model for the metabolic modification of behaviour. This study demonstrates that the activity of the small GTPase Rac2 is required in Drosophila for the proper regulation of lipid storage and feeding behaviour, as well as aggression and mating behaviours. Rac2 mutant males and females are susceptible to starvation and contain considerably less lipids than controls. Furthermore, Rac2 mutants also have disrupted feeding behaviour, eating fewer but larger meals than controls. Intriguingly, Rac2 mutant males rarely initiate aggressive behaviour and display significantly increased levels of courtship behaviour towards other males and mated females. From these results we conclude that Rac2 has a central role in regulating the Drosophila homeostatic system.

  19. Merkel Cell Polyomavirus Small T Antigen Drives Cell Motility via Rho-GTPase-Induced Filopodium Formation.

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    Stakaitytė, Gabrielė; Nwogu, Nnenna; Dobson, Samuel J; Knight, Laura M; Wasson, Christopher W; Salguero, Francisco J; Blackbourn, David J; Blair, G Eric; Mankouri, Jamel; Macdonald, Andrew; Whitehouse, Adrian

    2018-01-15

    Cell motility and migration is a complex, multistep, and multicomponent process intrinsic to progression and metastasis. Motility is dependent on the activities of integrin receptors and Rho family GTPases, resulting in the remodeling of the actin cytoskeleton and formation of various motile actin-based protrusions. Merkel cell carcinoma (MCC) is an aggressive skin cancer with a high likelihood of recurrence and metastasis. Merkel cell polyomavirus (MCPyV) is associated with the majority of MCC cases, and MCPyV-induced tumorigenesis largely depends on the expression of the small tumor antigen (ST). Since the discovery of MCPyV, a number of mechanisms have been suggested to account for replication and tumorigenesis, but to date, little is known about potential links between MCPyV T antigen expression and the metastatic nature of MCC. Previously, we described the action of MCPyV ST on the microtubule network and how it impacts cell motility and migration. Here, we demonstrate that MCPyV ST affects the actin cytoskeleton to promote the formation of filopodia through a mechanism involving the catalytic subunit of protein phosphatase 4 (PP4C). We also show that MCPyV ST-induced cell motility is dependent upon the activities of the Rho family GTPases Cdc42 and RhoA. In addition, our results indicate that the MCPyV ST-PP4C interaction results in the dephosphorylation of β 1 integrin, likely driving the cell motility pathway. These findings describe a novel mechanism by which a tumor virus induces cell motility, which may ultimately lead to cancer metastasis, and provides opportunities and strategies for targeted interventions for disseminated MCC. IMPORTANCE Merkel cell polyomavirus (MCPyV) is the most recently discovered human tumor virus. It causes the majority of cases of Merkel cell carcinoma (MCC), an aggressive skin cancer. However, the molecular mechanisms implicating MCPyV-encoded proteins in cancer development are yet to be fully elucidated. This study builds

  20. Identification of potential small molecule binding pockets on Rho family GTPases.

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    Juan Manuel Ortiz-Sanchez

    Full Text Available Rho GTPases are conformational switches that control a wide variety of signaling pathways critical for eukaryotic cell development and proliferation. They represent attractive targets for drug design as their aberrant function and deregulated activity is associated with many human diseases including cancer. Extensive high-resolution structures (>100 and recent mutagenesis studies have laid the foundation for the design of new structure-based chemotherapeutic strategies. Although the inhibition of Rho signaling with drug-like compounds is an active area of current research, very little attention has been devoted to directly inhibiting Rho by targeting potential allosteric non-nucleotide binding sites. By avoiding the nucleotide binding site, compounds may minimize the potential for undesirable off-target interactions with other ubiquitous GTP and ATP binding proteins. Here we describe the application of molecular dynamics simulations, principal component analysis, sequence conservation analysis, and ensemble small-molecule fragment mapping to provide an extensive mapping of potential small-molecule binding pockets on Rho family members. Characterized sites include novel pockets in the vicinity of the conformationaly responsive switch regions as well as distal sites that appear to be related to the conformations of the nucleotide binding region. Furthermore the use of accelerated molecular dynamics simulation, an advanced sampling method that extends the accessible time-scale of conventional simulations, is found to enhance the characterization of novel binding sites when conformational changes are important for the protein mechanism.

  1. Crosstalk between Bcl-2 family and Ras family small GTPases: potential cell fate regulation?

    International Nuclear Information System (INIS)

    Kang, Jia; Pervaiz, Shazib

    2013-01-01

    Cell fate regulation is a function of diverse cell signaling pathways that promote cell survival and or inhibit cell death execution. In this regard, the role of the Bcl-2 family in maintaining a tight balance between cell death and cell proliferation has been extensively studied. The conventional dogma links cell fate regulation by the Bcl-2 family to its effect on mitochondrial permeabilization and apoptosis amplification. However, recent evidence provide a novel mechanism for death regulation by the Bcl-2 family via modulating cellular redox metabolism. For example overexpression of Bcl-2 has been shown to contribute to a pro-oxidant intracellular milieu and down-regulation of cellular superoxide levels enhanced death sensitivity of Bcl-2 overexpressing cells. Interestingly, gene knockdown of the small GTPase Rac1 or pharmacological inhibition of its activity also reverted death phenotype in Bcl-2 expressing cells. This appears to be a function of an interaction between Bcl-2 and Rac1. Similar functional associations have been described between the Bcl-2 family and other members of the Ras superfamily. These interactions at the mitochondria provide novel opportunities for strategic therapeutic targeting of drug-resistant cancers.

  2. Epidermal activation of the small GTPase Rac1 in psoriasis pathogenesis.

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    Winge, Mårten C G; Marinkovich, M Peter

    2017-01-05

    The small GTPase Ras-related C3 botulinum toxin substrate 1 (RAC1) plays a central role in skin homeostasis, including barrier function, wound healing and inflammatory responses. Psoriasis is a common skin disease characterized by deregulation of these functions, and affected skin exhibit keratinocyte hyperproliferation, inflammation and immune cell infiltration. Although psoriasis is often triggered by environmental stimulus, there is a strong genetic association with genes expressed in both immune cells and keratinocytes, of which several are linked to Rac1 signaling. Rac1 is highly active in human psoriatic lesional skin and keratinocytes, and keratinocyte-specific overexpression of an activated mutant of Rac1, Rac1 V12 , in a transgenic mouse model closely mimics the presentation of human psoriasis. Both Rac1 activation in keratinocytes and immune derived stimulus are required to drive psoriasiform signaling in transgenic mouse and human xenograft models of psoriasis. Therefore, understanding how increased Rac1 activation in psoriatic epidermis is regulated is central to understanding how the abnormal crosstalk between keratinocytes and immune cells is maintained.

  3. Mechanisms of membrane binding of small GTPase K-Ras4B farnesylated hypervariable region.

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    Jang, Hyunbum; Abraham, Sherwin J; Chavan, Tanmay S; Hitchinson, Ben; Khavrutskii, Lyuba; Tarasova, Nadya I; Nussinov, Ruth; Gaponenko, Vadim

    2015-04-10

    K-Ras4B belongs to a family of small GTPases that regulates cell growth, differentiation and survival. K-ras is frequently mutated in cancer. K-Ras4B association with the plasma membrane through its farnesylated and positively charged C-terminal hypervariable region (HVR) is critical to its oncogenic function. However, the structural mechanisms of membrane association are not fully understood. Here, using confocal microscopy, surface plasmon resonance, and molecular dynamics simulations, we observed that K-Ras4B can be distributed in rigid and loosely packed membrane domains. Its membrane binding domain interaction with phospholipids is driven by membrane fluidity. The farnesyl group spontaneously inserts into the disordered lipid microdomains, whereas the rigid microdomains restrict the farnesyl group penetration. We speculate that the resulting farnesyl protrusion toward the cell interior allows oligomerization of the K-Ras4B membrane binding domain in rigid microdomains. Unlike other Ras isoforms, K-Ras4B HVR contains a single farnesyl modification and positively charged polylysine sequence. The high positive charge not only modulates specific HVR binding to anionic phospholipids but farnesyl membrane orientation. Phosphorylation of Ser-181 prohibits spontaneous farnesyl membrane insertion. The mechanism illuminates the roles of HVR modifications in K-Ras4B targeting microdomains of the plasma membrane and suggests an additional function for HVR in regulation of Ras signaling. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  4. Mechanisms of Membrane Binding of Small GTPase K-Ras4B Farnesylated Hypervariable Region*

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    Jang, Hyunbum; Abraham, Sherwin J.; Chavan, Tanmay S.; Hitchinson, Ben; Khavrutskii, Lyuba; Tarasova, Nadya I.; Nussinov, Ruth; Gaponenko, Vadim

    2015-01-01

    K-Ras4B belongs to a family of small GTPases that regulates cell growth, differentiation and survival. K-ras is frequently mutated in cancer. K-Ras4B association with the plasma membrane through its farnesylated and positively charged C-terminal hypervariable region (HVR) is critical to its oncogenic function. However, the structural mechanisms of membrane association are not fully understood. Here, using confocal microscopy, surface plasmon resonance, and molecular dynamics simulations, we observed that K-Ras4B can be distributed in rigid and loosely packed membrane domains. Its membrane binding domain interaction with phospholipids is driven by membrane fluidity. The farnesyl group spontaneously inserts into the disordered lipid microdomains, whereas the rigid microdomains restrict the farnesyl group penetration. We speculate that the resulting farnesyl protrusion toward the cell interior allows oligomerization of the K-Ras4B membrane binding domain in rigid microdomains. Unlike other Ras isoforms, K-Ras4B HVR contains a single farnesyl modification and positively charged polylysine sequence. The high positive charge not only modulates specific HVR binding to anionic phospholipids but farnesyl membrane orientation. Phosphorylation of Ser-181 prohibits spontaneous farnesyl membrane insertion. The mechanism illuminates the roles of HVR modifications in K-Ras4B targeting microdomains of the plasma membrane and suggests an additional function for HVR in regulation of Ras signaling. PMID:25713064

  5. The small GTPase Arl8b regulates assembly of the mammalian HOPS complex on lysosomes

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    Khatter, Divya; Raina, Vivek B.; Dwivedi, Devashish; Sindhwani, Aastha; Bahl, Surbhi; Sharma, Mahak

    2015-01-01

    The homotypic fusion and protein sorting (HOPS) complex is a multi-subunit complex conserved from yeast to mammals that regulates late endosome and lysosome fusion. However, little is known about how the HOPS complex is recruited to lysosomes in mammalian cells. Here, we report that the small GTPase Arl8b, but not Rab7 (also known as RAB7A), is essential for membrane localization of the human (h)Vps41 subunit of the HOPS complex. Assembly of the core HOPS subunits to Arl8b- and hVps41-positive lysosomes is guided by their subunit–subunit interactions. RNA interference (RNAi)-mediated depletion of hVps41 resulted in the impaired degradation of EGFR that was rescued upon expression of wild-type but not an Arl8b-binding-defective mutant of hVps41, suggesting that Arl8b-dependent lysosomal localization of hVps41 is required for its endocytic function. Furthermore, we have also identified that the Arl8b effector SKIP (also known as PLEKHM2) interacts with and recruits HOPS subunits to Arl8b and kinesin-positive peripheral lysosomes. Accordingly, RNAi-mediated depletion of SKIP impaired lysosomal trafficking and degradation of EGFR. These findings reveal that Arl8b regulates the association of the human HOPS complex with lysosomal membranes, which is crucial for the function of this tethering complex in endocytic degradation. PMID:25908847

  6. The small GTPase Arl8b regulates assembly of the mammalian HOPS complex on lysosomes.

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    Khatter, Divya; Raina, Vivek B; Dwivedi, Devashish; Sindhwani, Aastha; Bahl, Surbhi; Sharma, Mahak

    2015-05-01

    The homotypic fusion and protein sorting (HOPS) complex is a multi-subunit complex conserved from yeast to mammals that regulates late endosome and lysosome fusion. However, little is known about how the HOPS complex is recruited to lysosomes in mammalian cells. Here, we report that the small GTPase Arl8b, but not Rab7 (also known as RAB7A), is essential for membrane localization of the human (h)Vps41 subunit of the HOPS complex. Assembly of the core HOPS subunits to Arl8b- and hVps41-positive lysosomes is guided by their subunit-subunit interactions. RNA interference (RNAi)-mediated depletion of hVps41 resulted in the impaired degradation of EGFR that was rescued upon expression of wild-type but not an Arl8b-binding-defective mutant of hVps41, suggesting that Arl8b-dependent lysosomal localization of hVps41 is required for its endocytic function. Furthermore, we have also identified that the Arl8b effector SKIP (also known as PLEKHM2) interacts with and recruits HOPS subunits to Arl8b and kinesin-positive peripheral lysosomes. Accordingly, RNAi-mediated depletion of SKIP impaired lysosomal trafficking and degradation of EGFR. These findings reveal that Arl8b regulates the association of the human HOPS complex with lysosomal membranes, which is crucial for the function of this tethering complex in endocytic degradation. © 2015. Published by The Company of Biologists Ltd.

  7. Stage-specific functions of the small Rho GTPases Cdc42 and Rac1 for adult hippocampal neurogenesis

    DEFF Research Database (Denmark)

    Vadodaria, Krishna C; Brakebusch, Cord; Suter, Ueli

    2013-01-01

    The molecular mechanisms underlying the generation, maturation, and integration of new granule cells generated throughout life in the mammalian hippocampus remain poorly understood. Small Rho GTPases, such as Cdc42 and Rac1, have been implicated previously in neural stem/progenitor cell (NSPC......) proliferation and neuronal maturation during embryonic development. Here we used conditional genetic deletion and virus-based loss-of-function approaches to identify temporally distinct functions for Cdc42 and Rac1 in adult hippocampal neurogenesis. We found that Cdc42 is involved in mouse NSPC proliferation......, initial dendritic development, and dendritic spine maturation. In contrast, Rac1 is dispensable for early steps of neuronal development but is important for late steps of dendritic growth and spine maturation. These results establish cell-autonomous and stage-specific functions for the small Rho GTPases...

  8. Spatio-temporal manipulation of small GTPase activity at subcellular level and on timescale of seconds in living cells.

    Science.gov (United States)

    DeRose, Robert; Pohlmeyer, Christopher; Umeda, Nobuhiro; Ueno, Tasuku; Nagano, Tetsuo; Kuo, Scot; Inoue, Takanari

    2012-03-09

    Dynamic regulation of the Rho family of small guanosine triphosphatases (GTPases) with great spatiotemporal precision is essential for various cellular functions and events(1, 2). Their spatiotemporally dynamic nature has been revealed by visualization of their activity and localization in real time(3). In order to gain deeper understanding of their roles in diverse cellular functions at the molecular level, the next step should be perturbation of protein activities at a precise subcellular location and timing. To achieve this goal, we have developed a method for light-induced, spatio-temporally controlled activation of small GTPases by combining two techniques: (1) rapamycin-induced FKBP-FRB heterodimerization and (2) a photo-caging method of rapamycin. With the use of rapamycin-mediated FKBP-FRB heterodimerization, we have developed a method for rapidly inducible activation or inactivation of small GTPases including Rac(4), Cdc42(4), RhoA(4) and Ras(5), in which rapamycin induces translocation of FKBP-fused GTPases, or their activators, to the plasma membrane where FRB is anchored. For coupling with this heterodimerization system, we have also developed a photo-caging system of rapamycin analogs. A photo-caged compound is a small molecule whose activity is suppressed with a photocleavable protecting group known as a caging group. To suppress heterodimerization activity completely, we designed a caged rapamycin that is tethered to a macromolecule such that the resulting large complex cannot cross the plasma membrane, leading to virtually no background activity as a chemical dimerizer inside cells(6). Figure 1 illustrates a scheme of our system. With the combination of these two systems, we locally recruited a Rac activator to the plasma membrane on a timescale of seconds and achieved light-induced Rac activation at the subcellular level(6).

  9. Small GTPase R-Ras participates in neural tube formation in zebrafish embryonic spinal cord.

    Science.gov (United States)

    Ohata, Shinya; Uga, Hideko; Okamoto, Hitoshi; Katada, Toshiaki

    2018-06-27

    Ras related (R-Ras), a small GTPase, is involved in the maintenance of apico-basal polarity in neuroepithelial cells of the zebrafish hindbrain, axonal collapse in cultured murine hippocampal neurons, and maturation of blood vessels in adult mice. However, the role of R-Ras in neural tube formation remains unknown. Using antisense morpholino oligonucleotides (AMOs), we found that in the spinal cord of zebrafish embryos, the lumen was formed bilaterally in rras morphants, whereas it was formed at the midline in control embryos. As AMO can cause off-target effects, we generated rras mutant zebrafish lines using CRISPR/Cas9 technology. Although these rras mutant embryos did not have a bilateral lumen in the spinal cord, the following findings suggest that the phenotype is unlikely due to an off-target effect of rras AMO: 1) The rras morphant phenotype was rescued by an injection of AMO-resistant rras mRNA, and 2) a bilaterally segregated spinal cord was not observed in rras mutant embryos injected with rras AMO. The results suggest that the function of other ras family genes may be redundant in rras mutants. Previous research reported a bilaterally formed lumen in the spinal cord of zebrafish embryos with a mutation in a planar cell polarity (PCP) gene, van gogh-like 2 (vangl2). In the present study, in cultured cells, R-Ras was co-immunoprecipitated with Vangl2 but not with another PCP regulator, Pricke1. Interestingly, the interaction between R-Ras and Vangl2 was stronger in guanine-nucleotide free point mutants of R-Ras than in wild-type or constitutively active (GTP-bound) forms of R-Ras. R-Ras may regulate neural tube formation in cooperation with Vangl2 in the developing zebrafish spinal cord. Copyright © 2018 Elsevier Inc. All rights reserved.

  10. Nucleophosmin1 is a negative regulator of the small GTPase Rac1

    NARCIS (Netherlands)

    Zoughlami, Younes; van Stalborgh, Anne M.; van Hennik, Paula B.; Hordijk, Peter L.

    2013-01-01

    The Rac1 GTPase is a critical regulator of cytoskeletal dynamics and controls many biological processes, such as cell migration, cell-cell contacts, cellular growth and cell division. These complex processes are controlled by Rac1 signaling through effector proteins. We have previously identified

  11. The small GTPase RhoH is an atypical regulator of haematopoietic cells

    Directory of Open Access Journals (Sweden)

    Kubatzky Katharina F

    2008-09-01

    Full Text Available Abstract Rho GTPases are a distinct subfamily of the superfamily of Ras GTPases. The best-characterised members are RhoA, Rac and Cdc42 that regulate many diverse actions such as actin cytoskeleton reorganisation, adhesion, motility as well as cell proliferation, differentiation and gene transcription. Among the 20 members of that family, only Rac2 and RhoH show an expression restricted to the haematopoietic lineage. RhoH was first discovered in 1995 as a fusion transcript with the transcriptional repressor LAZ3/BCL6. It was therefore initially named translation three four (TTF but later on renamed RhoH due to its close relationship to the Ras/Rho family of GTPases. Since then, RhoH has been implicated in human cancer as the gene is subject to somatic hypermutation and by the detection of RHOH as a translocation partner for LAZ3/BCL6 or other genes in human lymphomas. Underexpression of RhoH is found in hairy cell leukaemia and acute myeloid leukaemia. Some of the amino acids that are crucial for GTPase activity are mutated in RhoH so that the protein is a GTPase-deficient, so-called atypical Rho GTPase. Therefore other mechanisms of regulating RhoH activity have been described. These include regulation at the mRNA level and tyrosine phosphorylation of the protein's unique ITAM-like motif. The C-terminal CaaX box of RhoH is mainly a target for farnesyl-transferase but can also be modified by geranylgeranyl-transferase. Isoprenylation of RhoH and changes in subcellular localisation may be an additional factor to fine-tune signalling. Little is currently known about its signalling, regulation or interaction partners. Recent studies have shown that RhoH negatively influences the proliferation and homing of murine haematopoietic progenitor cells, presumably by acting as an antagonist for Rac1. In leukocytes, RhoH is needed to keep the cells in a resting, non-adhesive state, but the exact mechanism has yet to be elucidated. RhoH has also been

  12. Stage-specific control of neural crest stem cell proliferation by the small rho GTPases Cdc42 and Rac1

    DEFF Research Database (Denmark)

    Fuchs, Sebastian; Herzog, Dominik; Sumara, Grzegorz

    2009-01-01

    -renewal and proliferation of later stage, but not early migratory NCSCs. This stage-specific requirement for small Rho GTPases is due to changes in NCSCs that, during development, acquire responsiveness to mitogenic EGF acting upstream of both Cdc42 and Rac1. Thus, our data reveal distinct mechanisms for growth control......The neural crest (NC) generates a variety of neural and non-neural tissues during vertebrate development. Both migratory NC cells and their target structures contain cells with stem cell features. Here we show that these populations of neural crest-derived stem cells (NCSCs) are differentially...

  13. Leucine-rich repeat kinase-1 regulates osteoclast function by modulating RAC1/Cdc42 Small GTPase phosphorylation and activation.

    Science.gov (United States)

    Zeng, Canjun; Goodluck, Helen; Qin, Xuezhong; Liu, Bo; Mohan, Subburaman; Xing, Weirong

    2016-10-01

    Leucine-rich repeat kinase-1 (Lrrk1) consists of ankyrin repeats (ANK), leucine-rich repeats (LRR), a GTPase-like domain of Roc (ROC), a COR domain, a serine/threonine kinase domain (KD), and WD40 repeats (WD40). Previous studies have revealed that knockout (KO) of Lrrk1 in mice causes severe osteopetrosis, and a human mutation of Lrrk1 leads to osteosclerotic metaphysial dysplasia. The molecular mechanism by which Lrrk1 regulates osteoclast function is unknown. In this study, we generated a series of Lrrk1 mutants and evaluated their ability to rescue defective bone resorption in Lrrk1-deficient osteoclasts by use of pit formation assays. Overexpression of Lrrk1 or LRR-truncated Lrrk1, but not ANK-truncated Lrrk1, WD40-truncated Lrrk1, Lrrk1-KD, or K651A mutant Lrrk1, rescued bone resorption function of Lrrk1 KO osteoclasts. We next examined whether RAC1/Cdc42 small GTPases are direct substrates of Lrrk1 in osteoclasts. Western blot and pull-down assays revealed that Lrrk1 deficiency in osteoclasts resulted in reduced phosphorylation and activation of RAC1/Cdc42. In vitro kinase assays confirmed that recombinant Lrrk1 phosphorylated RAC1-GST protein, and immunoprecipitation showed that the interaction of Lrrk1 with RAC1 occurred within 10 min after RANKL treatment. Overexpression of constitutively active Q61L RAC1 partially rescued the resorptive function of Lrrk1-deficient osteoclasts. Furthermore, lack of Lrrk1 in osteoclasts led to reduced autophosphorylation of p21 protein-activated kinase-1 at Ser 144 , catalyzed by RAC1/Cdc42 binding and activation. Our data indicate that Lrrk1 regulates osteoclast function by directly modulating phosphorylation and activation of small GTPase RAC1/Cdc42 and that its function depends on ANK, ROC, WD40, and kinase domains. Copyright © 2016 the American Physiological Society.

  14. Inhibition of endothelial cell proliferation by targeting Rac1 GTPase with small interference RNA in tumor cells

    International Nuclear Information System (INIS)

    Xue Yan; Bi Feng; Zhang Xueyong; Pan Yanglin; Liu Na; Zheng Yi; Fan Daiming

    2004-01-01

    Hypoxia-induced angiogenesis plays an important role in the malignancy of solid tumors. A number of recent studies including our own have suggested that Rho family small GTPases are involved in this process, and Rac1, a prominent member of the Rho family, may be critical in regulating hypoxia-induced gene activation of several angiogenesis factors and tumor suppressors. To further define Rac1 function in angiogenesis and to explore novel approaches to modulate angiogenesis, we employed the small interference RNA technique to knock down gene expression of Rac1 in gastric cancer cell line AGS that expresses a high level of Rac1. Both the mRNA and protein levels of Rac1 in the AGS cells were decreased dramatically after transfection with a Rac1-specific siRNA vector. When the conditioned medium derived from the Rac1 downregulated AGS cells was applied to the human endothelial cells, it could significantly inhibit the cell proliferation. Further study proved that, VEGF and HIF-1α, two angiogenesis promoting factors, were found to be downregulated whereas p53 and VHL, which are tumor suppressors and angiogenesis inhibitors, were upregulated in the Rac1 siRNA transfected cells. Our results suggest that Rac1 may be involved in angiogenesis by controlling the expression of angiogenesis-related factors and provide a possible strategy for the treatment of tumor angiogenesis by targeting the Rac1 GTPase

  15. Sevoflurane Inhalation Accelerates the Long-Term Memory Consolidation via Small GTPase Overexpression in the Hippocampus of Mice in Adolescence.

    Science.gov (United States)

    Nakamura, Emi; Kinoshita, Hiroyuki; Feng, Guo-Gang; Hayashi, Hisaki; Satomoto, Maiko; Sato, Motohiko; Fujiwara, Yoshihiro

    2016-01-01

    Sevoflurane exposure impairs the long-term memory in neonates. Whether the exposure to animals in adolescence affects the memory, however, has been unclear. A small hydrolase enzyme of guanosine triphosphate (GTPase) rac1 plays a role in the F-actin dynamics related to the synaptic plasticity, as well as superoxide production via reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activation. The current study was designed to examine whether sevoflurane exposure to mice in early adolescence modifies the long-term learning ability concomitantly with the changes in F-actin constitution as well as superoxide production in the hippocampus according to the levels of rac1 protein expression. Four-week-old mice were subjected to the evaluation of long-term learning ability for three days. On day one, each mouse was allowed to enter a dark chamber for five min to acclimatization. On day two, the procedure was repeated with the addition of an electric shock as soon as a mouse entered the dark chamber. All mice subsequently inhaled 2 L/min air with (Sevoflurane group) and without (Control group) 2.5% sevoflurane for three hours. On day three, each mouse was placed on the platform and retention time, which is the latency to enter the dark chamber, was examined. The brain removed after the behavior test, was used for analyses of immunofluorescence, Western immunoblotting and intracellular levels of superoxide. Sevoflurane exposure significantly prolonged retention time, indicating the enhanced long-term memory. Sevoflurane inhalation augmented F-actin constitution coexisting with the rac1 protein overexpression in the hippocampus whereas it did not alter the levels of superoxide. Sevoflurane exposure to 4-week-old mice accelerates the long-term memory concomitantly with the enhanced F-actin constitution coexisting with the small GTPase rac1 overexpression in the hippocampus. These results suggest that sevoflurane inhalation may amplify long-term memory

  16. The small GTPase Cdc42 modulates the number of exocytosis-competent dense-core vesicles in PC12 cells

    Energy Technology Data Exchange (ETDEWEB)

    Sato, Mai [Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro, Tokyo 153-8902 (Japan); Kitaguchi, Tetsuya [Cell Signaling Group, Waseda Bioscience Research Institute in Singapore (WABOIS), Waseda University, 11 Biopolis Way, 05-01/02 Helios, Singapore 138667 (Singapore); Numano, Rika [The Electronics-Inspired Interdisciplinary Research Institute (EIIRIS), Toyohashi University of Technology, 1-1 Hibarigaoka, Tennpaku-cho, Toyohashi, Aichi 441-8580 (Japan); Ikematsu, Kazuya [Forensic Pathology and Science, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki 852-8523 (Japan); Kakeyama, Masaki [Laboratory of Environmental Health Sciences, Center for Disease Biology and Integrative Medicine, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo 113-0033 (Japan); Murata, Masayuki; Sato, Ken [Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro, Tokyo 153-8902 (Japan); Tsuboi, Takashi, E-mail: takatsuboi@bio.c.u-tokyo.ac.jp [Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro, Tokyo 153-8902 (Japan)

    2012-04-06

    Highlights: Black-Right-Pointing-Pointer Regulation of exocytosis by Rho GTPase Cdc42. Black-Right-Pointing-Pointer Cdc42 increases the number of fusion events from newly recruited vesicles. Black-Right-Pointing-Pointer Cdc42 increases the number of exocytosis-competent dense-core vesicles. -- Abstract: Although the small GTPase Rho family Cdc42 has been shown to facilitate exocytosis through increasing the amount of hormones released, the precise mechanisms regulating the quantity of hormones released on exocytosis are not well understood. Here we show by live cell imaging analysis under TIRF microscope and immunocytochemical analysis under confocal microscope that Cdc42 modulated the number of fusion events and the number of dense-core vesicles produced in the cells. Overexpression of a wild-type or constitutively-active form of Cdc42 strongly facilitated high-KCl-induced exocytosis from the newly recruited plasma membrane vesicles in PC12 cells. By contrast, a dominant-negative form of Cdc42 inhibited exocytosis from both the newly recruited and previously docked plasma membrane vesicles. The number of intracellular dense-core vesicles was increased by the overexpression of both a wild-type and constitutively-active form of Cdc42. Consistently, activation of Cdc42 by overexpression of Tuba, a Golgi-associated guanine nucleotide exchange factor for Cdc42 increased the number of intracellular dense-core vesicles, whereas inhibition of Cdc42 by overexpression of the Cdc42/Rac interactive binding domain of neuronal Wiskott-Aldrich syndrome protein decreased the number of them. These findings suggest that Cdc42 facilitates exocytosis by modulating both the number of exocytosis-competent dense-core vesicles and the production of dense-core vesicles in PC12 cells.

  17. The small GTPase Cdc42 modulates the number of exocytosis-competent dense-core vesicles in PC12 cells

    International Nuclear Information System (INIS)

    Sato, Mai; Kitaguchi, Tetsuya; Numano, Rika; Ikematsu, Kazuya; Kakeyama, Masaki; Murata, Masayuki; Sato, Ken; Tsuboi, Takashi

    2012-01-01

    Highlights: ► Regulation of exocytosis by Rho GTPase Cdc42. ► Cdc42 increases the number of fusion events from newly recruited vesicles. ► Cdc42 increases the number of exocytosis-competent dense-core vesicles. -- Abstract: Although the small GTPase Rho family Cdc42 has been shown to facilitate exocytosis through increasing the amount of hormones released, the precise mechanisms regulating the quantity of hormones released on exocytosis are not well understood. Here we show by live cell imaging analysis under TIRF microscope and immunocytochemical analysis under confocal microscope that Cdc42 modulated the number of fusion events and the number of dense-core vesicles produced in the cells. Overexpression of a wild-type or constitutively-active form of Cdc42 strongly facilitated high-KCl-induced exocytosis from the newly recruited plasma membrane vesicles in PC12 cells. By contrast, a dominant-negative form of Cdc42 inhibited exocytosis from both the newly recruited and previously docked plasma membrane vesicles. The number of intracellular dense-core vesicles was increased by the overexpression of both a wild-type and constitutively-active form of Cdc42. Consistently, activation of Cdc42 by overexpression of Tuba, a Golgi-associated guanine nucleotide exchange factor for Cdc42 increased the number of intracellular dense-core vesicles, whereas inhibition of Cdc42 by overexpression of the Cdc42/Rac interactive binding domain of neuronal Wiskott–Aldrich syndrome protein decreased the number of them. These findings suggest that Cdc42 facilitates exocytosis by modulating both the number of exocytosis-competent dense-core vesicles and the production of dense-core vesicles in PC12 cells.

  18. Interaction of the Small GTPase Cdc42 with Arginine Kinase Restricts White Spot Syndrome Virus in Shrimp.

    Science.gov (United States)

    Xu, Ji-Dong; Jiang, Hai-Shan; Wei, Tian-Di; Zhang, Ke-Yi; Wang, Xian-Wei; Zhao, Xiao-Fan; Wang, Jin-Xing

    2017-03-01

    Many types of small GTPases are widely expressed in eukaryotes and have different functions. As a crucial member of the Rho GTPase family, Cdc42 serves a number of functions, such as regulating cell growth, migration, and cell movement. Several RNA viruses employ Cdc42-hijacking tactics in their target cell entry processes. However, the function of Cdc42 in shrimp antiviral immunity is not clear. In this study, we identified a Cdc42 protein in the kuruma shrimp ( Marsupenaeus japonicus ) and named it Mj Cdc42. Mj Cdc42 was upregulated in shrimp challenged by white spot syndrome virus (WSSV). The knockdown of Mj Cdc42 and injection of Cdc42 inhibitors increased the proliferation of WSSV. Further experiments determined that Mj Cdc42 interacted with an arginine kinase ( Mj AK). By analyzing the binding activity and enzyme activity of Mj AK and its mutant, Δ Mj AK, we found that Mj AK could enhance the replication of WSSV in shrimp. Mj AK interacted with the envelope protein VP26 of WSSV. An inhibitor of AK activity, quercetin, could impair the function of Mj AK in WSSV replication. Further study demonstrated that the binding of Mj Cdc42 and Mj AK depends on Cys 271 of Mj AK and suppresses the WSSV replication-promoting effect of Mj AK. By interacting with the active site of Mj AK and suppressing its enzyme activity, Mj Cdc42 inhibits WSSV replication in shrimp. Our results demonstrate a new function of Cdc42 in the cellular defense against viral infection in addition to the regulation of actin and phagocytosis, which has been reported in previous studies. IMPORTANCE The interaction of Cdc42 with arginine kinase plays a crucial role in the host defense against WSSV infection. This study identifies a new mechanism of Cdc42 in innate immunity and enriches the knowledge of the antiviral innate immunity of invertebrates. Copyright © 2017 American Society for Microbiology.

  19. Catalysis of GTP hydrolysis by small GTPases at atomic detail by integration of X-ray crystallography, experimental, and theoretical IR spectroscopy.

    Science.gov (United States)

    Rudack, Till; Jenrich, Sarah; Brucker, Sven; Vetter, Ingrid R; Gerwert, Klaus; Kötting, Carsten

    2015-10-02

    Small GTPases regulate key processes in cells. Malfunction of their GTPase reaction by mutations is involved in severe diseases. Here, we compare the GTPase reaction of the slower hydrolyzing GTPase Ran with Ras. By combination of time-resolved FTIR difference spectroscopy and QM/MM simulations we elucidate that the Mg(2+) coordination by the phosphate groups, which varies largely among the x-ray structures, is the same for Ran and Ras. A new x-ray structure of a Ran·RanBD1 complex with improved resolution confirmed this finding and revealed a general problem with the refinement of Mg(2+) in GTPases. The Mg(2+) coordination is not responsible for the much slower GTPase reaction of Ran. Instead, the location of the Tyr-39 side chain of Ran between the γ-phosphate and Gln-69 prevents the optimal positioning of the attacking water molecule by the Gln-69 relative to the γ-phosphate. This is confirmed in the RanY39A·RanBD1 crystal structure. The QM/MM simulations provide IR spectra of the catalytic center, which agree very nicely with the experimental ones. The combination of both methods can correlate spectra with structure at atomic detail. For example the FTIR difference spectra of RasA18T and RanT25A mutants show that spectral differences are mainly due to the hydrogen bond of Thr-25 to the α-phosphate in Ran. By integration of x-ray structure analysis, experimental, and theoretical IR spectroscopy the catalytic center of the x-ray structural models are further refined to sub-Å resolution, allowing an improved understanding of catalysis. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  20. Catalysis of GTP Hydrolysis by Small GTPases at Atomic Detail by Integration of X-ray Crystallography, Experimental, and Theoretical IR Spectroscopy*

    Science.gov (United States)

    Rudack, Till; Jenrich, Sarah; Brucker, Sven; Vetter, Ingrid R.; Gerwert, Klaus; Kötting, Carsten

    2015-01-01

    Small GTPases regulate key processes in cells. Malfunction of their GTPase reaction by mutations is involved in severe diseases. Here, we compare the GTPase reaction of the slower hydrolyzing GTPase Ran with Ras. By combination of time-resolved FTIR difference spectroscopy and QM/MM simulations we elucidate that the Mg2+ coordination by the phosphate groups, which varies largely among the x-ray structures, is the same for Ran and Ras. A new x-ray structure of a Ran·RanBD1 complex with improved resolution confirmed this finding and revealed a general problem with the refinement of Mg2+ in GTPases. The Mg2+ coordination is not responsible for the much slower GTPase reaction of Ran. Instead, the location of the Tyr-39 side chain of Ran between the γ-phosphate and Gln-69 prevents the optimal positioning of the attacking water molecule by the Gln-69 relative to the γ-phosphate. This is confirmed in the RanY39A·RanBD1 crystal structure. The QM/MM simulations provide IR spectra of the catalytic center, which agree very nicely with the experimental ones. The combination of both methods can correlate spectra with structure at atomic detail. For example the FTIR difference spectra of RasA18T and RanT25A mutants show that spectral differences are mainly due to the hydrogen bond of Thr-25 to the α-phosphate in Ran. By integration of x-ray structure analysis, experimental, and theoretical IR spectroscopy the catalytic center of the x-ray structural models are further refined to sub-Å resolution, allowing an improved understanding of catalysis. PMID:26272610

  1. Different roles of the small GTPases Rac1, Cdc42, and RhoG in CALEB/NGC-induced dendritic tree complexity.

    Science.gov (United States)

    Schulz, Jana; Franke, Kristin; Frick, Manfred; Schumacher, Stefan

    2016-10-01

    Rho GTPases play prominent roles in the regulation of cytoskeletal reorganization. Many aspects have been elaborated concerning the individual functions of Rho GTPases in distinct signaling pathways leading to cytoskeletal rearrangements. However, major questions have yet to be answered regarding the integration and the signaling hierarchy of different Rho GTPases in regulating the cytoskeleton in fundamental physiological events like neuronal process differentiation. Here, we investigate the roles of the small GTPases Rac1, Cdc42, and RhoG in defining dendritic tree complexity stimulated by the transmembrane epidermal growth factor family member CALEB/NGC. Combining gain-of-function and loss-of-function analysis in primary hippocampal neurons, we find that Rac1 is essential for CALEB/NGC-mediated dendritic branching. Cdc42 reduces the complexity of dendritic trees. Interestingly, we identify the palmitoylated isoform of Cdc42 to adversely affect dendritic outgrowth and dendritic branching, whereas the prenylated Cdc42 isoform does not. In contrast to Rac1, CALEB/NGC and Cdc42 are not directly interconnected in regulating dendritic tree complexity. Unlike Rac1, the Rac1-related GTPase RhoG reduces the complexity of dendritic trees by acting upstream of CALEB/NGC. Mechanistically, CALEB/NGC activates Rac1, and RhoG reduces the amount of CALEB/NGC that is located at the right site for Rac1 activation at the cell membrane. Thus, Rac1, Cdc42, and RhoG perform very specific and non-redundant functions at different levels of hierarchy in regulating dendritic tree complexity induced by CALEB/NGC. Rho GTPases play a prominent role in dendritic branching. CALEB/NGC is a transmembrane member of the epidermal growth factor (EGF) family that mediates dendritic branching, dependent on Rac1. CALEB/NGC stimulates Rac1 activity. RhoG inhibits CALEB/NGC-mediated dendritic branching by decreasing the amount of CALEB/NGC at the plasma membrane. Palmitoylated, but not prenylated form

  2. Role of the Small GTPase Rho3 in Golgi/Endosome trafficking through functional interaction with adaptin in Fission Yeast.

    Directory of Open Access Journals (Sweden)

    Ayako Kita

    Full Text Available BACKGROUND: We had previously identified the mutant allele of apm1(+ that encodes a homolog of the mammalian µ1A subunit of the clathrin-associated adaptor protein-1 (AP-1 complex, and we demonstrated the role of Apm1 in Golgi/endosome trafficking, secretion, and vacuole fusion in fission yeast. METHODOLOGY/PRINCIPAL FINDINGS: In the present study, we isolated rho3(+, which encodes a Rho-family small GTPase, an important regulator of exocystosis, as a multicopy-suppressor of the temperature-sensitive growth of the apm1-1 mutant cells. Overexpression of Rho3 suppressed the Cl(- sensitivity and immunosuppressant sensitivity of the apm1-1 mutant cells. Overexpression of Rho3 also suppressed the fragmentation of vacuoles, and the accumulation of v-SNARE Syb1 in Golgi/endosomes and partially suppressed the defective secretion associated with apm1-deletion cells. Notably, electron microscopic observation of the rho3-deletion cells revealed the accumulation of abnormal Golgi-like structures, vacuole fragmentation, and accumulation of secretory vesicles; these phenotypes were very similar to those of the apm1-deletion cells. Furthermore, the rho3-deletion cells and apm1-deletion cells showed very similar phenotypic characteristics, including the sensitivity to the immunosuppressant FK506, the cell wall-damaging agent micafungin, Cl(-, and valproic acid. Green fluorescent protein (GFP-Rho3 was localized at Golgi/endosomes as well as the plasma membrane and division site. Finally, Rho3 was shown to form a complex with Apm1 as well as with other subunits of the clathrin-associated AP-1 complex in a GTP- and effector domain-dependent manner. CONCLUSIONS/SIGNIFICANCE: Taken together, our findings reveal a novel role of Rho3 in the regulation of Golgi/endosome trafficking and suggest that clathrin-associated adaptor protein-1 and Rho3 co-ordinate in intracellular transport in fission yeast. To the best of our knowledge, this study provides the first evidence

  3. The small GTPase Rac1 is required for smooth muscle contraction

    DEFF Research Database (Denmark)

    Rahman, Awahan; Davis, Benjamin; Lövdahl, Cecilia

    2014-01-01

    The role of the small GTP-binding protein Rac1 in smooth muscle contraction was examined using small molecule inhibitors (EHT1864, NSC23766) and a novel smooth muscle-specific, conditional, Rac1 knockout mouse strain. EHT1864, which affects nucleotide binding and inhibits Rac1 activity, concentra...

  4. Apical accumulation of the Sevenless receptor tyrosine kinase during Drosophila eye development is promoted by the small GTPase Rap1.

    Science.gov (United States)

    Baril, Caroline; Lefrançois, Martin; Sahmi, Malha; Knævelsrud, Helene; Therrien, Marc

    2014-08-01

    The Ras/MAPK-signaling pathway plays pivotal roles during development of metazoans by controlling cell proliferation and cell differentiation elicited, in several instances, by receptor tyrosine kinases (RTKs). While the internal mechanism of RTK-driven Ras/MAPK signaling is well understood, far less is known regarding its interplay with other co-required signaling events involved in developmental decisions. In a genetic screen designed to identify new regulators of RTK/Ras/MAPK signaling during Drosophila eye development, we identified the small GTPase Rap1, PDZ-GEF, and Canoe as components contributing to Ras/MAPK-mediated R7 cell differentiation. Rap1 signaling has recently been found to participate in assembling cadherin-based adherens junctions in various fly epithelial tissues. Here, we show that Rap1 activity is required for the integrity of the apical domains of developing photoreceptor cells and that reduced Rap1 signaling hampers the apical accumulation of the Sevenless RTK in presumptive R7 cells. It thus appears that, in addition to its role in cell-cell adhesion, Rap1 signaling controls the partitioning of the epithelial cell membrane, which in turn influences signaling events that rely on apico-basal cell polarity. Copyright © 2014 by the Genetics Society of America.

  5. The small Rho GTPase Rac1 controls normal human dermal fibroblasts proliferation with phosphorylation of the oncoprotein c-myc

    International Nuclear Information System (INIS)

    Nikolova, Ekaterina; Mitev, Vanio; Zhelev, Nikolai; Deroanne, Christophe F.; Poumay, Yves

    2007-01-01

    Proliferation of dermal fibroblasts is crucial for the maintenance of skin. The small Rho GTPase, Rac1, has been identified as a key transducer of proliferative signals in various cell types, but in normal human dermal fibroblasts its significance to cell growth control has not been studied. In this study, we applied the method of RNA interference to suppress endogenous Rac1 expression and examined the consequences on human skin fibroblasts. Rac1 knock-down resulted in inhibition of DNA synthesis. This effect was not mediated by inhibition of the central transducer of proliferative stimuli, ERK1/2 or by activation of the pro-apoptotic p38. Rather, as a consequence of the suppressed Rac1 expression we observed a significant decrease in phosphorylation of c-myc, revealing for the first time that in human fibroblasts Rac1 exerts control on proliferation through c-myc phosphorylation. Thus Rac1 activates proliferation of normal fibroblasts through stimulation of c-myc phosphorylation without affecting ERK1/2 activity

  6. Gain-of-function mutant p53 activates small GTPase Rac1 through SUMOylation to promote tumor progression.

    Science.gov (United States)

    Yue, Xuetian; Zhang, Cen; Zhao, Yuhan; Liu, Juan; Lin, Alan W; Tan, Victor M; Drake, Justin M; Liu, Lianxin; Boateng, Michael N; Li, Jun; Feng, Zhaohui; Hu, Wenwei

    2017-08-15

    Tumor suppressor p53 is frequently mutated in human cancer. Mutant p53 often promotes tumor progression through gain-of-function (GOF) mechanisms. However, the mechanisms underlying mutant p53 GOF are not well understood. In this study, we found that mutant p53 activates small GTPase Rac1 as a critical mechanism for mutant p53 GOF to promote tumor progression. Mechanistically, mutant p53 interacts with Rac1 and inhibits its interaction with SUMO-specific protease 1 (SENP1), which in turn inhibits SENP1-mediated de-SUMOylation of Rac1 to activate Rac1. Targeting Rac1 signaling by RNAi, expression of the dominant-negative Rac1 (Rac1 DN), or the specific Rac1 inhibitor NSC23766 greatly inhibits mutant p53 GOF in promoting tumor growth and metastasis. Furthermore, mutant p53 expression is associated with enhanced Rac1 activity in clinical tumor samples. These results uncover a new mechanism for Rac1 activation in tumors and, most importantly, reveal that activation of Rac1 is an unidentified and critical mechanism for mutant p53 GOF in tumorigenesis, which could be targeted for therapy in tumors containing mutant p53. © 2017 Yue et al.; Published by Cold Spring Harbor Laboratory Press.

  7. Glutaminase 2 is a novel negative regulator of small GTPase Rac1 and mediates p53 function in suppressing metastasis

    Science.gov (United States)

    Zhang, Cen; Liu, Juan; Zhao, Yuhan; Yue, Xuetian; Zhu, Yu; Wang, Xiaolong; Wu, Hao; Blanco, Felix; Li, Shaohua; Bhanot, Gyan; Haffty, Bruce G; Hu, Wenwei; Feng, Zhaohui

    2016-01-01

    Glutaminase (GLS) isoenzymes GLS1 and GLS2 are key enzymes for glutamine metabolism. Interestingly, GLS1 and GLS2 display contrasting functions in tumorigenesis with elusive mechanism; GLS1 promotes tumorigenesis, whereas GLS2 exhibits a tumor-suppressive function. In this study, we found that GLS2 but not GLS1 binds to small GTPase Rac1 and inhibits its interaction with Rac1 activators guanine-nucleotide exchange factors, which in turn inhibits Rac1 to suppress cancer metastasis. This function of GLS2 is independent of GLS2 glutaminase activity. Furthermore, decreased GLS2 expression is associated with enhanced metastasis in human cancer. As a p53 target, GLS2 mediates p53’s function in metastasis suppression through inhibiting Rac1. In summary, our results reveal that GLS2 is a novel negative regulator of Rac1, and uncover a novel function and mechanism whereby GLS2 suppresses metastasis. Our results also elucidate a novel mechanism that contributes to the contrasting functions of GLS1 and GLS2 in tumorigenesis. DOI: http://dx.doi.org/10.7554/eLife.10727.001 PMID:26751560

  8. Activation of the Small GTPase Rap1 Inhibits Choroidal Neovascularization by Regulating Cell Junctions and ROS Generation in Rats.

    Science.gov (United States)

    Li, Jiajia; Zhang, Rong; Wang, Caixia; Wang, Xin; Xu, Man; Ma, Jingxue; Shang, Qingli

    2018-03-30

    Choroidal neovascularization (CNV) is a common vision-threatening complication associated with many  fundus diseases. The retinal pigment epithelial (RPE) cell junction barrier has critical functions in preventing CNV, and oxidative stress can cause compromise of barrier integrity and induce angiogenesis. Rap1, a small guanosine triphosphatase (GTPase), is involved in regulating endothelial and epithelial cell junctions. In this work, we explored the function and mechanism of Rap1 in CNV in vivo. A laser-induced rat CNV model was developed. Rap1 was activated through intravitreal injection of the Rap1 activator 8CPT-2'-O-Me-cAMP (8CPT). At 14 days after laser treatment, CNV size in RPE/choroid flat mounts was measured by fluorescein isothiocyanate-dextran staining. Expression of vascular endothelial growth factor (VEGF) and cell junction proteins in RPE/choroid tissues were analyzed by western blots and quantitative real-time PCR assays. Reactive oxygen species (ROS) in RPE cells were detectedbydichloro-dihydro-fluorescein diacetate assays. The antioxidant apocynin was intraperitoneally injected into rats. Activating Rap1 by 8CPT significantly reduced CNV size and VEGF expression in the rat CNV model. Rap1 activation enhanced protein and mRNA levels of ZO-1 and occludin, two tight junction proteins in the RPE barrier. In addition, reducing ROS generation by injection of apocynin, a NADPH oxidase inhibitor, inhibited CNV formation. Rap1 activation reduced ROS generation and expression of NADPH oxidase 4. Rap1 activation inhibits CNV through regulating barrier integrity and ROS generation of RPE in vivo, and selectively activating Rap1 may be a way to reduce vision loss from CNV.

  9. Mutations in the small GTPase gene RAB39B are responsible for X-linked mental retardation associated with autism, epilepsy, and macrocephaly.

    Science.gov (United States)

    Giannandrea, Maila; Bianchi, Veronica; Mignogna, Maria Lidia; Sirri, Alessandra; Carrabino, Salvatore; D'Elia, Errico; Vecellio, Matteo; Russo, Silvia; Cogliati, Francesca; Larizza, Lidia; Ropers, Hans-Hilger; Tzschach, Andreas; Kalscheuer, Vera; Oehl-Jaschkowitz, Barbara; Skinner, Cindy; Schwartz, Charles E; Gecz, Jozef; Van Esch, Hilde; Raynaud, Martine; Chelly, Jamel; de Brouwer, Arjan P M; Toniolo, Daniela; D'Adamo, Patrizia

    2010-02-12

    Human Mental Retardation (MR) is a common and highly heterogeneous pediatric disorder affecting around 3% of the general population; at least 215 X-linked MR (XLMR) conditions have been described, and mutations have been identified in 83 different genes, encoding proteins with a variety of function, such as chromatin remodeling, synaptic function, and intracellular trafficking. The small GTPases of the RAB family, which play an essential role in intracellular vesicular trafficking, have been shown to be involved in MR. We report here the identification of mutations in the small GTPase RAB39B gene in two male patients. One mutation in family X (D-23) introduced a stop codon seven amino acids after the start codon (c.21C > A; p.Y7X). A second mutation, in the MRX72 family, altered the 5' splice site (c.215+1G > A) and normal splicing. Neither instance produced a protein. Mutations segregate with the disease in the families, and in some family members intellectual disabilities were associated with autism spectrum disorder, epileptic seizures, and macrocephaly. We show that RAB39B, a novel RAB GTPase of unknown function, is a neuronal-specific protein that is localized to the Golgi compartment. Its downregulation leads to an alteration in the number and morphology of neurite growth cones and a significant reduction in presynaptic buttons, suggesting that RAB39B is required for synapse formation and maintenance. Our results demonstrate developmental and functional neuronal alteration as a consequence of downregulation of RAB39B and emphasize the critical role of vesicular trafficking in the development of neurons and human intellectual abilities. Copyright (c) 2010 The American Society of Human Genetics. Published by Elsevier Inc. All rights reserved.

  10. Enrichment of Phosphatidylethanolamine in Viral Replication Compartments via Co-opting the Endosomal Rab5 Small GTPase by a Positive-Strand RNA Virus.

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    Kai Xu

    2016-10-01

    Full Text Available Positive-strand RNA viruses build extensive membranous replication compartments to support replication and protect the virus from antiviral responses by the host. These viruses require host factors and various lipids to form viral replication complexes (VRCs. The VRCs built by Tomato bushy stunt virus (TBSV are enriched with phosphatidylethanolamine (PE through a previously unknown pathway. To unravel the mechanism of PE enrichment within the TBSV replication compartment, in this paper, the authors demonstrate that TBSV co-opts the guanosine triphosphate (GTP-bound active form of the endosomal Rab5 small GTPase via direct interaction with the viral replication protein. Deletion of Rab5 orthologs in a yeast model host or expression of dominant negative mutants of plant Rab5 greatly decreases TBSV replication and prevents the redistribution of PE to the sites of viral replication. We also show that enrichment of PE in the viral replication compartment is assisted by actin filaments. Interestingly, the closely related Carnation Italian ringspot virus, which replicates on the boundary membrane of mitochondria, uses a similar strategy to the peroxisomal TBSV to hijack the Rab5-positive endosomes into the viral replication compartments. Altogether, usurping the GTP-Rab5-positive endosomes allows TBSV to build a PE-enriched viral replication compartment, which is needed to support peak-level replication. Thus, the Rab family of small GTPases includes critical host factors assisting VRC assembly and genesis of the viral replication compartment.

  11. Tumor endothelial marker 5 expression in endothelial cells during capillary morphogenesis is induced by the small GTPase Rac and mediates contact inhibition of cell proliferation

    Energy Technology Data Exchange (ETDEWEB)

    Vallon, Mario, E-mail: m.vallon@arcor.de [Nuklearmedizinische Klinik und Poliklinik, Technische Universitaet Muenchen, Ismaninger Strasse 22, 81675 Munich (Germany); Rohde, Franziska; Janssen, Klaus-Peter [Chirurgische Klinik und Poliklinik, Technische Universitaet Muenchen, Munich (Germany); Essler, Markus [Nuklearmedizinische Klinik und Poliklinik, Technische Universitaet Muenchen, Ismaninger Strasse 22, 81675 Munich (Germany)

    2010-02-01

    Tumor endothelial marker (TEM) 5 is an adhesion G-protein-coupled receptor upregulated in endothelial cells during tumor and physiologic angiogenesis. So far, the mechanisms leading to upregulation of TEM5 and its function during angiogenesis have not been identified. Here, we report that TEM5 expression in endothelial cells is induced during capillary-like network formation on Matrigel, during capillary morphogenesis in a three-dimensional collagen I matrix, and upon confluence on a two-dimensional matrix. TEM5 expression was not induced by a variety of soluble angiogenic factors, including VEGF and bFGF, in subconfluent endothelial cells. TEM5 upregulation was blocked by toxin B from Clostridium difficile, an inhibitor of the small GTPases Rho, Rac, and Cdc42. The Rho inhibitor C3 transferase from Clostridium botulinum did not affect TEM5 expression, whereas the Rac inhibitor NSC23766 suppressed TEM5 upregulation. An excess of the soluble TEM5 extracellular domain or an inhibitory monoclonal TEM5 antibody blocked contact inhibition of endothelial cell proliferation resulting in multilayered islands within the endothelial monolayer and increased vessel density during capillary formation. Based on our results we conclude that TEM5 expression during capillary morphogenesis is induced by the small GTPase Rac and mediates contact inhibition of proliferation in endothelial cells.

  12. The Tip of the Four N-Terminal α-Helices of Clostridium sordellii Lethal Toxin Contains the Interaction Site with Membrane Phosphatidylserine Facilitating Small GTPases Glucosylation

    Directory of Open Access Journals (Sweden)

    Carolina Varela Chavez

    2016-03-01

    Full Text Available Clostridium sordellii lethal toxin (TcsL is a powerful virulence factor responsible for severe toxic shock in man and animals. TcsL belongs to the large clostridial glucosylating toxin (LCGT family which inactivates small GTPases by glucosylation with uridine-diphosphate (UDP-glucose as a cofactor. Notably, TcsL modifies Rac and Ras GTPases, leading to drastic alteration of the actin cytoskeleton and cell viability. TcsL enters cells via receptor-mediated endocytosis and delivers the N-terminal glucosylating domain (TcsL-cat into the cytosol. TcsL-cat was found to preferentially bind to phosphatidylserine (PS-containing membranes and to increase the glucosylation of Rac anchored to the lipid membrane. We have previously reported that the N-terminal four helical bundle structure (1–93 domain recognizes a broad range of lipids, but that TcsL-cat specifically binds to PS and phosphatidic acid. Here, we show using mutagenesis that the PS binding site is localized on the tip of the four-helix bundle which is rich in positively-charged amino acids. Residues Y14, V15, F17, and R18 on loop 1, between helices 1 and 2, in coordination with R68 from loop 3, between helices 3 and 4, form a pocket which accommodates L-serine. The functional PS-binding site is required for TcsL-cat binding to the plasma membrane and subsequent cytotoxicity. TcsL-cat binding to PS facilitates a high enzymatic activity towards membrane-anchored Ras by about three orders of magnitude as compared to Ras in solution. The PS-binding site is conserved in LCGTs, which likely retain a common mechanism of binding to the membrane for their full activity towards membrane-bound GTPases.

  13. A Pan-GTPase Inhibitor as a Molecular Probe.

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    Lin Hong

    Full Text Available Overactive GTPases have often been linked to human diseases. The available inhibitors are limited and have not progressed far in clinical trials. We report here a first-in-class small molecule pan-GTPase inhibitor discovered from a high throughput screening campaign. The compound CID1067700 inhibits multiple GTPases in biochemical, cellular protein and protein interaction, as well as cellular functional assays. In the biochemical and protein interaction assays, representative GTPases from Rho, Ras, and Rab, the three most generic subfamilies of the GTPases, were probed, while in the functional assays, physiological processes regulated by each of the three subfamilies of the GTPases were examined. The chemical functionalities essential for the activity of the compound were identified through structural derivatization. The compound is validated as a useful molecular probe upon which GTPase-targeting inhibitors with drug potentials might be developed.

  14. Two Components of Aversive Memory in Drosophila, Anesthesia-Sensitive and Anesthesia-Resistant Memory, Require Distinct Domains Within the Rgk1 Small GTPase.

    Science.gov (United States)

    Murakami, Satoshi; Minami-Ohtsubo, Maki; Nakato, Ryuichiro; Shirahige, Katsuhiko; Tabata, Tetsuya

    2017-05-31

    Multiple components have been identified that exhibit different stabilities for aversive olfactory memory in Drosophila These components have been defined by behavioral and genetic studies and genes specifically required for a specific component have also been identified. Intermediate-term memory generated after single cycle conditioning is divided into anesthesia-sensitive memory (ASM) and anesthesia-resistant memory (ARM), with the latter being more stable. We determined that the ASM and ARM pathways converged on the Rgk1 small GTPase and that the N-terminal domain-deleted Rgk1 was sufficient for ASM formation, whereas the full-length form was required for ARM formation. Rgk1 is specifically accumulated at the synaptic site of the Kenyon cells (KCs), the intrinsic neurons of the mushroom bodies, which play a pivotal role in olfactory memory formation. A higher than normal Rgk1 level enhanced memory retention, which is consistent with the result that Rgk1 suppressed Rac-dependent memory decay; these findings suggest that rgk1 bolsters ASM via the suppression of forgetting. We propose that Rgk1 plays a pivotal role in the regulation of memory stabilization by serving as a molecular node that resides at KC synapses, where the ASM and ARM pathway may interact. SIGNIFICANCE STATEMENT Memory consists of multiple components. Drosophila olfactory memory serves as a fundamental model with which to investigate the mechanisms that underlie memory formation and has provided genetic and molecular means to identify the components of memory, namely short-term, intermediate-term, and long-term memory, depending on how long the memory lasts. Intermediate memory is further divided into anesthesia-sensitive memory (ASM) and anesthesia-resistant memory (ARM), with the latter being more stable. We have identified a small GTPase in Drosophila , Rgk1, which plays a pivotal role in the regulation of olfactory memory stability. Rgk1 is required for both ASM and ARM. Moreover, N

  15. MDA-9/Syntenin (SDCBP) modulates small GTPases RhoA and Cdc42 via transforming growth factor β1 to enhance epithelial-mesenchymal transition in breast cancer.

    Science.gov (United States)

    Menezes, Mitchell E; Shen, Xue-Ning; Das, Swadesh K; Emdad, Luni; Sarkar, Devanand; Fisher, Paul B

    2016-12-06

    Epithelial-mesenchymal transition (EMT) is one of the decisive steps regulating cancer invasion and metastasis. However, the molecular mechanisms underlying this transition require further clarification. MDA-9/syntenin (SDCBP) expression is elevated in breast cancer patient samples as well as cultured breast cancer cells. Silencing expression of MDA-9 in mesenchymal metastatic breast cancer cells triggered a change in cell morphology in both 2D- and 3D-cultures to a more epithelial-like phenotype, along with changes in EMT markers, cytoskeletal rearrangement and decreased invasion. Conversely, over expressing MDA-9 in epithelial non-metastatic breast cancer cells instigated a change in morphology to a more mesenchymal phenotype with corresponding changes in EMT markers, cytoskeletal rearrangement and an increase in invasion. We also found that MDA-9 upregulated active levels of known modulators of EMT, the small GTPases RhoA and Cdc42, via TGFβ1. Reintroducing TGFβ1 in MDA-9 silenced cells restored active RhoA and cdc42 levels, modulated cytoskeletal rearrangement and increased invasion. We further determined that MDA-9 interacts with TGFβ1 via its PDZ1 domain. Finally, in vivo studies demonstrated that silencing the expression of MDA-9 resulted in decreased lung metastasis and TGFβ1 re-expression partially restored lung metastases. Our findings provide evidence for the relevance of MDA-9 in mediating EMT in breast cancer and support the potential of MDA-9 as a therapeutic target against metastatic disease.

  16. Ras-dva1 small GTPase regulates telencephalon development in Xenopus laevis embryos by controlling Fgf8 and Agr signaling at the anterior border of the neural plate

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    Maria B. Tereshina

    2014-07-01

    Full Text Available We previously found that the small GTPase Ras-dva1 is essential for the telencephalic development in Xenopus laevis because Ras-dva1 controls the Fgf8-mediated induction of FoxG1 expression, a key telencephalic regulator. In this report, we show, however, that Ras-dva1 and FoxG1 are expressed in different groups of cells; whereas Ras-dva1 is expressed in the outer layer of the anterior neural fold, FoxG1 and Fgf8 are activated in the inner layer from which the telencephalon is derived. We resolve this paradox by demonstrating that Ras-dva1 is involved in the transduction of Fgf8 signal received by cells in the outer layer, which in turn send a feedback signal that stimulates FoxG1 expression in the inner layer. We show that this feedback signal is transmitted by secreted Agr proteins, the expression of which is activated in the outer layer by mediation of Ras-dva1 and the homeodomain transcription factor Otx2. In turn, Agrs are essential for maintaining Fgf8 and FoxG1 expression in cells at the anterior neural plate border. Our finding reveals a novel feedback loop mechanism based on the exchange of Fgf8 and Agr signaling between neural and non-neural compartments at the anterior margin of the neural plate and demonstrates a key role of Ras-dva1 in this mechanism.

  17. 2D-DIGE and MALDI TOF/TOF MS analysis reveal that small GTPase signaling pathways may play an important role in cadmium-induced colon cell malignant transformation

    International Nuclear Information System (INIS)

    Lu, Jian; Zhou, Zhongping; Zheng, Jianzhou; Zhang, Zhuyi; Lu, Rongzhu; Liu, Hanqing; Shi, Haifeng; Tu, Zhigang

    2015-01-01

    Cadmium is a toxic heavy metal present in the environment and in industrial materials. Cadmium has demonstrated carcinogenic activity that induces cell transformation, but how this occurs is unclear. We used 2D-DIGE and MALDI TOF/TOF MS combined with bioinformatics and immunoblotting to investigate the molecular mechanism of cadmium transformation. We found that small GTPases were critical for transformation. Additionally, proteins involved in mitochondrial transcription, DNA repair, and translation also had altered expression patterns in cadmium treated cells. Collectively, our results suggest that activation of small GTPases contributes to cadmium-induced transformation of colon cells. - Highlights: • Colon epithelial cell line is firstly successfully transformed by cadmium. • 2D-DIGE is applied to visualize the differentially expressed proteins. • RhoA plays an important role in cadmium induced malignant transformation. • Bioinformatic and experimental methods are combined to explore new mechanisms.

  18. 2D-DIGE and MALDI TOF/TOF MS analysis reveal that small GTPase signaling pathways may play an important role in cadmium-induced colon cell malignant transformation

    Energy Technology Data Exchange (ETDEWEB)

    Lu, Jian, E-mail: lujian@ujs.edu.cn [School of Medicine, Jiangsu University, Zhenjiang 212013 (China); Institute of Life Sciences, Jiangsu University, Zhenjiang 212013 (China); Zhou, Zhongping [School of Medicine, Jiangsu University, Zhenjiang 212013 (China); Institute of Life Sciences, Jiangsu University, Zhenjiang 212013 (China); Zheng, Jianzhou [Department of Respiration Medicine, Changzhou No.2 People' s Hospital, Changzhou 213003 (China); Zhang, Zhuyi [School of Medicine, Jiangsu University, Zhenjiang 212013 (China); Institute of Life Sciences, Jiangsu University, Zhenjiang 212013 (China); Lu, Rongzhu [School of Medicine, Jiangsu University, Zhenjiang 212013 (China); Liu, Hanqing [School of Pharmacy, Jiangsu University, Zhenjiang 212013 (China); Shi, Haifeng [Institute of Life Sciences, Jiangsu University, Zhenjiang 212013 (China); Tu, Zhigang, E-mail: tuzg_ujs@ujs.edu.cn [Institute of Life Sciences, Jiangsu University, Zhenjiang 212013 (China)

    2015-10-01

    Cadmium is a toxic heavy metal present in the environment and in industrial materials. Cadmium has demonstrated carcinogenic activity that induces cell transformation, but how this occurs is unclear. We used 2D-DIGE and MALDI TOF/TOF MS combined with bioinformatics and immunoblotting to investigate the molecular mechanism of cadmium transformation. We found that small GTPases were critical for transformation. Additionally, proteins involved in mitochondrial transcription, DNA repair, and translation also had altered expression patterns in cadmium treated cells. Collectively, our results suggest that activation of small GTPases contributes to cadmium-induced transformation of colon cells. - Highlights: • Colon epithelial cell line is firstly successfully transformed by cadmium. • 2D-DIGE is applied to visualize the differentially expressed proteins. • RhoA plays an important role in cadmium induced malignant transformation. • Bioinformatic and experimental methods are combined to explore new mechanisms.

  19. Rho GTPases in ameloblast differentiation

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    Keishi Otsu

    2016-05-01

    Full Text Available During tooth development, ameloblasts differentiate from inner enamel epithelial cells to enamel-forming cells by modulating the signal pathways mediating epithelial–mesenchymal interaction and a cell-autonomous gene network. The differentiation process of epithelial cells is characterized by marked changes in their morphology and polarity, accompanied by dynamic cytoskeletal reorganization and changes in cell–cell and cell–matrix adhesion over time. Functional ameloblasts are tall, columnar, polarized cells that synthesize and secrete enamel-specific proteins. After deposition of the full thickness of enamel matrix, ameloblasts become smaller and regulate enamel maturation. Recent significant advances in the fields of molecular biology and genetics have improved our understanding of the regulatory mechanism of the ameloblast cell life cycle, mediated by the Rho family of small GTPases. They act as intracellular molecular switch that transduce signals from extracellular stimuli to the actin cytoskeleton and the nucleus. In our review, we summarize studies that provide current evidence for Rho GTPases and their involvement in ameloblast differentiation. In addition to the Rho GTPases themselves, their downstream effectors and upstream regulators have also been implicated in ameloblast differentiation.

  20. Coordinated regulation by two VPS9 domain-containing guanine nucleotide exchange factors in small GTPase Rab5 signaling pathways in fission yeast

    International Nuclear Information System (INIS)

    Tsukamoto, Yuta; Kagiwada, Satoshi; Shimazu, Sayuri; Takegawa, Kaoru; Noguchi, Tetsuko; Miyamoto, Masaaki

    2015-01-01

    The small GTPase Rab5 is reported to regulate various cellular functions, such as vesicular transport and endocytosis. VPS9 domain-containing proteins are thought to activate Rab5(s) by their guanine-nucleotide exchange activities. Numerous VPS9 proteins have been identified and are structurally conserved from yeast to mammalian cells. However, the functional relationships among VPS9 proteins in cells remain unclear. Only one Rab5 and two VPS9 proteins were identified in the Schizosaccharomyces pombe genome. Here, we examined the cellular function of two VPS9 proteins and the relationship between these proteins in cellular functions. Vps901-GFP and Vps902-GFP exhibited dotted signals in vegetative and differentiated cells. vps901 deletion mutant (Δvps901) cells exhibited a phenotype deficient in the mating process and responses to high concentrations of ions, such as calcium and metals, and Δvps901Δvps902 double mutant cells exhibited round cell shapes similar to ypt5-909 (Rab5 mutant allele) cells. Deletion of both vps901 and vps902 genes completely abolished the mating process and responses to various stresses. A lack of vacuole formation and aberrant inner cell membrane structures were also observed in Δvps901Δvps902 cells by electron microscopy. These data strongly suggest that Vps901 and Vps902 are cooperatively involved in the regulation of cellular functions, such as cell morphology, sexual development, response to ion stresses, and vacuole formation, via Rab5 signaling pathways in fission yeast cells. - Highlights: • Roles of Rab5 activator VPS9 proteins in cellular functions. • Cooperation between VPS9 proteins in Rab5 signaling pathway. • Roles of each VPS9 protein in Rab5 signaling pathway are discussed

  1. Coordinated regulation by two VPS9 domain-containing guanine nucleotide exchange factors in small GTPase Rab5 signaling pathways in fission yeast

    Energy Technology Data Exchange (ETDEWEB)

    Tsukamoto, Yuta [Department of Biology, Graduate School of Science, Kobe University, 1-1 Rokkodai-cho, Nada, Kobe 657-8501 (Japan); Kagiwada, Satoshi [Department of Biological Sciences, Faculty of Science, Nara Women' s University, Kitauoyanishi-machi, Nara 630-8506 (Japan); Shimazu, Sayuri [Center for Supports to Research and Education Activities, Kobe University, 1-1 Rokkodai-cho, Nada, Kobe 657-8501 (Japan); Takegawa, Kaoru [Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581 (Japan); Noguchi, Tetsuko [Department of Biological Sciences, Faculty of Science, Nara Women' s University, Kitauoyanishi-machi, Nara 630-8506 (Japan); Miyamoto, Masaaki, E-mail: miya@kobe-u.ac.jp [Department of Biology, Graduate School of Science, Kobe University, 1-1 Rokkodai-cho, Nada, Kobe 657-8501 (Japan); Center for Supports to Research and Education Activities, Kobe University, 1-1 Rokkodai-cho, Nada, Kobe 657-8501 (Japan)

    2015-03-20

    The small GTPase Rab5 is reported to regulate various cellular functions, such as vesicular transport and endocytosis. VPS9 domain-containing proteins are thought to activate Rab5(s) by their guanine-nucleotide exchange activities. Numerous VPS9 proteins have been identified and are structurally conserved from yeast to mammalian cells. However, the functional relationships among VPS9 proteins in cells remain unclear. Only one Rab5 and two VPS9 proteins were identified in the Schizosaccharomyces pombe genome. Here, we examined the cellular function of two VPS9 proteins and the relationship between these proteins in cellular functions. Vps901-GFP and Vps902-GFP exhibited dotted signals in vegetative and differentiated cells. vps901 deletion mutant (Δvps901) cells exhibited a phenotype deficient in the mating process and responses to high concentrations of ions, such as calcium and metals, and Δvps901Δvps902 double mutant cells exhibited round cell shapes similar to ypt5-909 (Rab5 mutant allele) cells. Deletion of both vps901 and vps902 genes completely abolished the mating process and responses to various stresses. A lack of vacuole formation and aberrant inner cell membrane structures were also observed in Δvps901Δvps902 cells by electron microscopy. These data strongly suggest that Vps901 and Vps902 are cooperatively involved in the regulation of cellular functions, such as cell morphology, sexual development, response to ion stresses, and vacuole formation, via Rab5 signaling pathways in fission yeast cells. - Highlights: • Roles of Rab5 activator VPS9 proteins in cellular functions. • Cooperation between VPS9 proteins in Rab5 signaling pathway. • Roles of each VPS9 protein in Rab5 signaling pathway are discussed.

  2. Commercial scale irradiation for insect disinfestation preserves peach quality

    International Nuclear Information System (INIS)

    McDonald, Heather; McCulloch, Mary; Caporaso, Fred; Winborne, Ian; Oubichon, Michon; Rakovski, Cyril; Prakash, Anuradha

    2012-01-01

    Irradiation is approved as a generic quarantine treatment by the US Department of Agriculture, Animal and Plant Health Inspection Service. Due to the effectiveness of irradiation in controlling insects on commodities, there is a growing need to understand the effects of low dose irradiation on fruit quality. The goal of this study was to determine the sensitivity of peaches (Prunus persica) to irradiation, and secondly, to determine the effect of commercial scale treatment on shelf-life, overall quality and consumer liking. Six varieties of peaches were irradiated in small batches at 0.29, 0.49, 0.69 and 0.90 kGy to observe the sensitivity of peaches at different dose levels. Changes in quality were evaluated by 8 trained panelists using descriptive analysis. Sensory characteristics (color, smoothness, aroma, touch firmness, mouth firmness, graininess, overall flavor and off-flavor) were evaluated at 2–4 day intervals and untreated samples served as control. To simulate commercial treatment, peaches were irradiated in pallet quantities at a target dose level of 0.4 kGy. The average absorbed dose was 0.66 kGy with an average dose uniformity ratio of 1.57. Commercially treated peaches were evaluated by 40–80 untrained consumers for acceptability routinely throughout the shelf life. Titratable acidity, Brix, texture and weight loss were also monitored for both commercial and small scale irradiated peaches. There was no dose effect on TA, Brix and weight loss due to irradiation. Peaches irradiated at 0.69 and 0.90 kGy were darker in flesh color, more juicy and less firm as determined by the trained panel and analytical pressure tests. Commercial scale irradiation did not adversely affect shelf life but was seen to enhance ripening. This, however, was perceived as a positive change by consumers. Overall, consumers rated the acceptability of irradiated peaches higher than untreated peaches. Statistical analysis was performed using linear mixed models to find

  3. GTPases and the origin of the ribosome

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    Smith Temple F

    2010-05-01

    Full Text Available Abstract Background This paper is an attempt to trace the evolution of the ribosome through the evolution of the universal P-loop GTPases that are involved with the ribosome in translation and with the attachment of the ribosome to the membrane. The GTPases involved in translation in Bacteria/Archaea are the elongation factors EFTu/EF1, the initiation factors IF2/aeIF5b + aeIF2, and the elongation factors EFG/EF2. All of these GTPases also contain the OB fold also found in the non GTPase IF1 involved in initiation. The GTPase involved in the signal recognition particle in most Bacteria and Archaea is SRP54. Results 1 The Elongation Factors of the Archaea based on structural considerations of the domains have the following evolutionary path: EF1→ aeIF2 → EF2. The evolution of the aeIF5b was a later event; 2 the Elongation Factors of the Bacteria based on the topological considerations of the GTPase domain have a similar evolutionary path: EFTu→ IF→2→EFG. These evolutionary sequences reflect the evolution of the LSU followed by the SSU to form the ribosome; 3 the OB-fold IF1 is a mimic of an ancient tRNA minihelix. Conclusion The evolution of translational GTPases of both the Archaea and Bacteria point to the evolution of the ribosome. The elongation factors, EFTu/EF1, began as a Ras-like GTPase bringing the activated minihelix tRNA to the Large Subunit Unit. The initiation factors and elongation factor would then have evolved from the EFTu/EF1 as the small subunit was added to the evolving ribosome. The SRP has an SRP54 GTPase and a specific RNA fold in its RNA component similar to the PTC. We consider the SRP to be a remnant of an ancient form of an LSU bound to a membrane. Reviewers This article was reviewed by George Fox, Leonid Mirny and Chris Sander.

  4. The small GTPase Rab5 homologue Ypt5 regulates cell morphology, sexual development, ion-stress response and vacuolar formation in fission yeast

    Energy Technology Data Exchange (ETDEWEB)

    Tsukamoto, Yuta; Katayama, Chisako [Graduate School of Science, Kobe University, 1-1 Rokkodai-cho Nada, Kobe 657-8501 (Japan); Shinohara, Miki; Shinohara, Akira [Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871 (Japan); Maekawa, Shohei [Graduate School of Science, Kobe University, 1-1 Rokkodai-cho Nada, Kobe 657-8501 (Japan); Miyamoto, Masaaki, E-mail: miya@kobe-u.ac.jp [Graduate School of Science, Kobe University, 1-1 Rokkodai-cho Nada, Kobe 657-8501 (Japan); Center for Supports to Research and Education Activities, Kobe University, 1-1 Rokkodai-cho Nada, Kobe 657-8501 (Japan)

    2013-11-29

    Highlights: •Multiple functions of Rab5 GTPase in fission yeast were found. •Roles of Rab5 in fission yeast were discussed. •Relation between Rab5 and actin cytoskeleton were discussed. -- Abstract: Inner-membrane transport is critical to cell function. Rab family GTPases play an important role in vesicle transport. In mammalian cells, Rab5 is reported to be involved in the regulation of endosome formation, phagocytosis and chromosome alignment. Here, we examined the role of the fission yeast Rab5 homologue Ypt5 using a point mutant allele. Mutant cells displayed abnormal cell morphology, mating, sporulation, endocytosis, vacuole fusion and responses to ion stress. Our data strongly suggest that fission yeast Rab5 is involved in the regulation of various types of cellular functions.

  5. The small GTPase Rab5 homologue Ypt5 regulates cell morphology, sexual development, ion-stress response and vacuolar formation in fission yeast

    International Nuclear Information System (INIS)

    Tsukamoto, Yuta; Katayama, Chisako; Shinohara, Miki; Shinohara, Akira; Maekawa, Shohei; Miyamoto, Masaaki

    2013-01-01

    Highlights: •Multiple functions of Rab5 GTPase in fission yeast were found. •Roles of Rab5 in fission yeast were discussed. •Relation between Rab5 and actin cytoskeleton were discussed. -- Abstract: Inner-membrane transport is critical to cell function. Rab family GTPases play an important role in vesicle transport. In mammalian cells, Rab5 is reported to be involved in the regulation of endosome formation, phagocytosis and chromosome alignment. Here, we examined the role of the fission yeast Rab5 homologue Ypt5 using a point mutant allele. Mutant cells displayed abnormal cell morphology, mating, sporulation, endocytosis, vacuole fusion and responses to ion stress. Our data strongly suggest that fission yeast Rab5 is involved in the regulation of various types of cellular functions

  6. In situ detection of the activation of Rac1 and RalA small GTPases in mouse adipocytes by immunofluorescent microscopy following in vivo and ex vivo insulin stimulation.

    Science.gov (United States)

    Takenaka, Nobuyuki; Nihata, Yuma; Ueda, Sho; Satoh, Takaya

    2017-11-01

    Rac1 has been implicated in insulin-dependent glucose uptake by mechanisms involving plasma membrane translocation of the glucose transporter GLUT4 in skeletal muscle. Although the uptake of glucose is also stimulated by insulin in adipose tissue, the role for Rac1 in adipocyte insulin signaling remains controversial. As a step to reveal the role for Rac1 in adipocytes, we aimed to establish immunofluorescent microscopy to detect the intracellular distribution of activated Rac1. The epitope-tagged Rac1-binding domain of a Rac1-specific target was utilized as a probe that specifically recognizes the activated form of Rac1. Rac1 activation in response to ex vivo and in vivo insulin stimulations in primary adipocyte culture and mouse white adipose tissue, respectively, was successfully observed by immunofluorescent microscopy. These Rac1 activations were mediated by phosphoinositide 3-kinase. Another small GTPase RalA has also been implicated in insulin-stimulated glucose uptake in skeletal muscle and adipose tissue. Similarly to Rac1, immunofluorescent microscopy using an activated RalA-specific polypeptide probe allowed us to detect intracellular distribution of insulin-activated RalA in adipocytes. These novel approaches to visualize the activation status of small GTPases in adipocytes will largely contribute to the understanding of signal transduction mechanisms particularly for insulin action. Copyright © 2017 Elsevier Inc. All rights reserved.

  7. Rho GTPase function in tumorigenesis

    DEFF Research Database (Denmark)

    Karlsson, R; Pedersen, Esben Ditlev Kølle; Wang, Zhipeng

    2009-01-01

    , for that reason, Rho GTPases, their regulators, and their effectors have been suggested to control tumor formation and progression in humans. However, while the tumor-relevant functions of Rho GTPases are very well documented in vitro, we are only now beginning to assess their contribution to cancer in human...... patients and in animal models. This review will give a very brief overview of Rho GTPase function in general and then focus on in vivo evidence for a role of Rho GTPases in malignant tumors, both in human patients and in genetically modified mice....

  8. Gallic acid inhibits gastric cancer cells metastasis and invasive growth via increased expression of RhoB, downregulation of AKT/small GTPase signals and inhibition of NF-κB activity

    Energy Technology Data Exchange (ETDEWEB)

    Ho, Hsieh-Hsun [Institute of Biochemistry and Biotechnology, Chung Shan Medical University, Taichung 402, Taiwan (China); Chang, Chi-Sen [Department of Medicine, Chung Shan Medical University, Taichung 402, Taiwan (China); Division of Gastroenterology, Taichung Veterans General Hospital, Taichung 402, Taiwan (China); Ho, Wei-Chi [Division of Gastroenterology, Jen-Ai Hospital, Taichung 402, Taiwan (China); Liao, Sheng-You [Institute of Biochemistry and Biotechnology, Chung Shan Medical University, Taichung 402, Taiwan (China); Lin, Wea-Lung [Department of Pathology, School of Medicine, Chung Shan Medical University, Taichung 402, Taiwan (China); Department of Pathology, Chung Shan Medical University Hospital, Taichung 402, Taiwan (China); Wang, Chau-Jong, E-mail: wcj@csmu.edu.tw [Institute of Biochemistry and Biotechnology, Chung Shan Medical University, Taichung 402, Taiwan (China); Department of Medical Research, Chung Shan Medical University Hospital, Taichung 402, Taiwan (China)

    2013-01-01

    Our previous study demonstrated the therapeutic potential of gallic acid (GA) for controlling tumor metastasis through its inhibitory effect on the motility of AGS cells. A noteworthy finding in our previous experiment was increased RhoB expression in GA-treated cells. The aim of this study was to evaluate the role of RhoB expression on the inhibitory effects of GA on AGS cells. By applying the transfection of RhoB siRNA into AGS cells and an animal model, we tested the effect of GA on inhibition of tumor growth and RhoB expression. The results confirmed that RhoB-siRNA transfection induced GA to inhibit AGS cells’ invasive growth involving blocking the AKT/small GTPase signals pathway and inhibition of NF-κB activity. Finally, we evaluated the effect of GA on AGS cell metastasis by colonization of tumor cells in nude mice. It showed GA inhibited tumor cells growth via the expression of RhoB. These data support the inhibitory effect of GA which was shown to inhibit gastric cancer cell metastasis and invasive growth via increased expression of RhoB, downregulation of AKT/small GTPase signals and inhibition of NF-κB activity. Thus, GA might be a potential agent in treating gastric cancer. Highlights: ► GA could downregulate AKT signal via increased expression of RhoB. ► GA inhibits metastasis in vitro in gastric carcinoma. ► GA inhibits tumor growth in nude mice model.

  9. The Small GTPase Rac1 Contributes to Extinction of Aversive Memories of Drug Withdrawal by Facilitating GABAA Receptor Endocytosis in the vmPFC.

    Science.gov (United States)

    Wang, Weisheng; Ju, Yun-Yue; Zhou, Qi-Xin; Tang, Jian-Xin; Li, Meng; Zhang, Lei; Kang, Shuo; Chen, Zhong-Guo; Wang, Yu-Jun; Ji, Hui; Ding, Yu-Qiang; Xu, Lin; Liu, Jing-Gen

    2017-07-26

    Extinction of aversive memories has been a major concern in neuropsychiatric disorders, such as anxiety disorders and drug addiction. However, the mechanisms underlying extinction of aversive memories are not fully understood. Here, we report that extinction of conditioned place aversion (CPA) to naloxone-precipitated opiate withdrawal in male rats activates Rho GTPase Rac1 in the ventromedial prefrontal cortex (vmPFC) in a BDNF-dependent manner, which determines GABA A receptor (GABA A R) endocytosis via triggering synaptic translocation of activity-regulated cytoskeleton-associated protein (Arc) through facilitating actin polymerization. Active Rac1 is essential and sufficient for GABA A R endocytosis and CPA extinction. Knockdown of Rac1 expression within the vmPFC of rats using Rac1-shRNA suppressed GABA A R endocytosis and CPA extinction, whereas expression of a constitutively active form of Rac1 accelerated GABA A R endocytosis and CPA extinction. The crucial role of GABA A R endocytosis in the LTP induction and CPA extinction is evinced by the findings that blockade of GABA A R endocytosis by a dynamin function-blocking peptide (Myr-P4) abolishes LTP induction and CPA extinction. Thus, the present study provides first evidence that Rac1-dependent GABA A R endocytosis plays a crucial role in extinction of aversive memories and reveals the sequence of molecular events that contribute to learning experience modulation of synaptic GABA A R endocytosis. SIGNIFICANCE STATEMENT This study reveals that Rac1-dependent GABA A R endocytosis plays a crucial role in extinction of aversive memories associated with drug withdrawal and identifies Arc as a downstream effector of Rac1 regulations of synaptic plasticity as well as learning and memory, thereby suggesting therapeutic targets to promote extinction of the unwanted memories. Copyright © 2017 the authors 0270-6474/17/377096-15$15.00/0.

  10. Rac and Rho GTPases in cancer cell motility control

    Directory of Open Access Journals (Sweden)

    Parri Matteo

    2010-09-01

    Full Text Available Abstract Rho GTPases represent a family of small GTP-binding proteins involved in cell cytoskeleton organization, migration, transcription, and proliferation. A common theme of these processes is a dynamic reorganization of actin cytoskeleton which has now emerged as a major switch control mainly carried out by Rho and Rac GTPase subfamilies, playing an acknowledged role in adaptation of cell motility to the microenvironment. Cells exhibit three distinct modes of migration when invading the 3 D environment. Collective motility leads to movement of cohorts of cells which maintain the adherens junctions and move by photolytic degradation of matrix barriers. Single cell mesenchymal-type movement is characterized by an elongated cellular shape and again requires extracellular proteolysis and integrin engagement. In addition it depends on Rac1-mediated cell polarization and lamellipodia formation. Conversely, in amoeboid movement cells have a rounded morphology, the movement is independent from proteases but requires high Rho GTPase to drive elevated levels of actomyosin contractility. These two modes of cell movement are interconvertible and several moving cells, including tumor cells, show an high degree of plasticity in motility styles shifting ad hoc between mesenchymal or amoeboid movements. This review will focus on the role of Rac and Rho small GTPases in cell motility and in the complex relationship driving the reciprocal control between Rac and Rho granting for the opportunistic motile behaviour of aggressive cancer cells. In addition we analyse the role of these GTPases in cancer progression and metastatic dissemination.

  11. Effects of different products of peach (Prunus persica L. Batsch) from a variety developed in southern Brazil on oxidative stress and inflammatory parameters in vitro and ex vivo

    Science.gov (United States)

    Gasparotto, Juciano; Somensi, Nauana; Bortolin, Rafael Calixto; Moresco, Karla Suzana; Girardi, Carolina Saibro; Klafke, Karina; Rabelo, Thallita Kelly; Morrone, Maurilio Da Silva; Vizzotto, Márcia; Raseira, Maria do Carmo Bassols; Moreira, José Claudio Fonseca; Gelain, Daniel Pens

    2014-01-01

    Antioxidant, anti-glycation and anti-inflammatory activities of fresh and conserved peach fruits (Prunus persica L. Batsch) were compared. Fresh peach pulps, peels, preserve peach pulps and the preserve syrup were prepared at equal concentrations. Rat liver, kidney and brain cortex tissue slices were pre-incubated with peach samples, subjected to oxidative stress with FeSO4 and hydrogen peroxide. Fresh peach pulps and peel conferred higher protection against cytotoxicity and oxidative stress than preserve peach pulps in most tissues. Release of tumor necrosis factor-α and interleukin-1β was also significantly decreased by Fresh peach pulps and peel, followed by preserve peach pulps. Total phenolic determination and HPLC analysis of carotenoids showed that the content of secondary metabolites in Fresh peach pulps and peel is significantly higher than in preserve peach pulps, while the syrup had only small or trace amounts of these compounds. Fresh peach pulps and Peel demonstrated high antioxidant and anti-inflammatory effects preventing against induced damage. PMID:25320458

  12. Mutation scanning of peach floral genes

    Directory of Open Access Journals (Sweden)

    Wilde H Dayton

    2011-05-01

    Full Text Available Abstract Background Mutation scanning technology has been used to develop crop species with improved traits. Modifications that improve screening throughput and sensitivity would facilitate the targeted mutation breeding of crops. Technical innovations for high-resolution melting (HRM analysis are enabling the clinic-based screening for human disease gene polymorphism. We examined the application of two HRM modifications, COLD-PCR and QMC-PCR, to the mutation scanning of genes in peach, Prunus persica. The targeted genes were the putative floral regulators PpAGAMOUS and PpTERMINAL FLOWER I. Results HRM analysis of PpAG and PpTFL1 coding regions in 36 peach cultivars found one polymorphic site in each gene. PpTFL1 and PpAG SNPs were used to examine approaches to increase HRM throughput. Cultivars with SNPs could be reliably detected in pools of twelve genotypes. COLD-PCR was found to increase the sensitivity of HRM analysis of pooled samples, but worked best with small amplicons. Examination of QMC-PCR demonstrated that primary PCR products for further analysis could be produced from variable levels of genomic DNA. Conclusions Natural SNPs in exons of target peach genes were discovered by HRM analysis of cultivars from a southeastern US breeding program. For detecting natural or induced SNPs in larger populations, HRM efficiency can be improved by increasing sample pooling and template production through approaches such as COLD-PCR and QMC-PCR. Technical advances developed to improve clinical diagnostics can play a role in the targeted mutation breeding of crops.

  13. Comprehensive behavioral analysis of mice deficient in Rapgef2 and Rapgef6, a subfamily of guanine nucleotide exchange factors for Rap small GTPases possessing the Ras/Rap-associating domain.

    Science.gov (United States)

    Maeta, Kazuhiro; Hattori, Satoko; Ikutomo, Junji; Edamatsu, Hironori; Bilasy, Shymaa E; Miyakawa, Tsuyoshi; Kataoka, Tohru

    2018-05-10

    Rapgef2 and Rapgef6 define a subfamily of guanine nucleotide exchange factors for Rap small GTPases, characterized by the possession of the Ras/Rap-associating domain. Previous genomic analyses suggested their possible involvement in the etiology of schizophrenia. We recently demonstrated the development of an ectopic cortical mass (ECM), which resembles the human subcortical band heterotopia, in the dorsal telencephalon-specific Rapgef2 conditional knockout (Rapgef2-cKO) brains. Additional knockout of Rapgef6 in Rapgef2-cKO mice resulted in gross enlargement of the ECM whereas knockout of Rapgef6 alone (Rapgef6-KO) had no discernible effect on the brain morphology. Here, we performed a battery of behavioral tests to examine the effects of Rapgef2 or Rapgef6 deficiency on higher brain functions. Rapgef2-cKO mice exhibited hyperlocomotion phenotypes. They showed decreased anxiety-like behavior in the elevated plus maze and the open-field tests as well as increased depression-like behavior in the Porsolt forced swim and tail suspension tests. They also exhibited increased sociability especially in novel environments. They showed defects in cognitive function as evidenced by reduced learning ability in the Barnes circular maze test and by impaired working memory in the T maze tests. In contrast, although Rapgef6 and Rapgef2 share similarities in biochemical roles, Rapgef6-KO mice exhibited mild behavioral abnormalities detected with a number of behavioral tests, such as hyperlocomotion phenotype in the open-field test and the social interaction test with a novel environment and working-memory defects in the T-maze test. In conclusion, although there were differences in their brain morphology and the magnitude of the behavioral abnormalities, Rapgef2-cKO mice and Rapgef6-KO mice exhibited hyperlocomotion phenotype and working-memory defect, both of which could be recognized as schizophrenia-like behavior.

  14. Role of Rab family GTPases and their effectors in melanosomal logistics.

    Science.gov (United States)

    Ohbayashi, Norihiko; Fukuda, Mitsunori

    2012-04-01

    Rab GTPases constitute a family of small GTPases that regulate a variety of membrane trafficking events in all eukaryotic cells by recruiting their specific effector molecules. Recent accumulating evidence indicates that members of the mammalian Rab small GTPase family are involved in certain physiological and pathological processes. In particular, functional impairments of specific Rab proteins, e.g. Rab38 and Rab27A, their regulators or their effectors cause pigmentation disorders in humans and coat colour variations in mice because such impairments cause defects in melanosomal logistics, i.e. defects in melanosome biogenesis and transport. Genetic and biochemical analyses of the gene products responsible for mammalian pigmentation disorders in the past decade have revealed that Rab-mediated endosomal transport systems and melanosome transport systems play crucial roles in the efficient darkening of mammalian hair and skin. In this article, we review current knowledge regarding melanosomal logistics, with particular focus on the roles of Rab small GTPases and their effectors.

  15. Susceptibility of peach GF 305 seedlings and selected herbaceous plants to plum pox virus isolates from western Slovakia.

    Science.gov (United States)

    Glasa, M; Matisová, J; Hricovský, I; Kúdela, O

    1997-12-01

    The susceptibility of peach GF 305 seedlings and herbaceous plants to five plum pox virus (PPV) isolates from orchards of western Slovakia was investigated. PPV was isolated from diseased plum, apricot and peach trees, and transmitted by chip-budding to peach GF 305. The herbaceous plants were infected by mechanical inoculation. The transmission was analysed by symptomatology and double sandwich enzyme-linked immunosorbent assay (DAS-ELISA). Infected peaches developed leaf distortion, tissue clearing along the veins and small chlorotic spots (isolate BOR-3). With exception of BOR-3, the PPV isolates transmitted from peach caused local chlorotic spots on Chenopodium foetidum. The character of symptoms changed when a sap from PPV-infected Nicotiana benthamiana was used as virus inoculum. From N. benthamiana, the PPV isolates could be transmitted to Pisum sativum, cv. Colmo (light green mosaic), N. clevelandii and N. clevelandii x N. glutinosa hybrid (latent infection or chlorotic spots).

  16. Physicochemical, antioxidant, and sensorial properties of peach ...

    African Journals Online (AJOL)

    Preparation of varieties of processed peach products could be a good option to supply in lean season. Objective of the study was to assess the quality characteristics of peach snacks prepared from 11 different cultivars. We investigated the physicochemical (soluble solid, titratable acidity, hardness and dry yield), antioxidant ...

  17. Peach Flower Monitoring Using Aerial Multispectral Imaging

    Directory of Open Access Journals (Sweden)

    Ryan Horton

    2017-01-01

    Full Text Available One of the tools for optimal crop production is regular monitoring and assessment of crops. During the growing season of fruit trees, the bloom period has increased photosynthetic rates that correlate with the fruiting process. This paper presents the development of an image processing algorithm to detect peach blossoms on trees. Aerial images of peach (Prunus persica trees were acquired from both experimental and commercial peach orchards in the southwestern part of Idaho using an off-the-shelf unmanned aerial system (UAS, equipped with a multispectral camera (near-infrared, green, blue. The image processing algorithm included contrast stretching of the three bands to enhance the image and thresholding segmentation method to detect the peach blossoms. Initial results showed that the image processing algorithm could detect peach blossoms with an average detection rate of 84.3% and demonstrated good potential as a monitoring tool for orchard management.

  18. Cryopreservation of peach palm zygotic embryos.

    Science.gov (United States)

    Steinmacher, Douglas A; Saldanha, Cleber W; Clement, Charles R; Guerra, Miguel P

    2007-01-01

    Cryopreservation is a safe and cost-effective option for long-term germplasm conservation of non-orthodox seed species, such as peach palm (Bactris gasipaes). The objective of the present study was to establish a cryopreservation protocol for peach palm zygotic embryos based on the encapsulation-dehydration technique. After excision, zygotic embryos were encapsulated with 3 percent sodium alginate plus 2 M glycerol and 0.4 M sucrose, and pre-treated or not with 1 M sucrose during 24 h, followed by air-drying. Fresh weight water contents of beads decreased from 83 percent and 87 percent to 18 percent and 20 percent for pre-treated or non-pretreated beads, respectively, after 4 h of dehydration. Sucrose pre-treatment at 1 M caused lower zygotic embryo germination and plantlet height in contrast to non-treated beads. All the variables were statistically influenced by dehydration time. Optimal conditions for recovery of cryopreserved zygotic embryos include encapsulation and dehydration for 4 h in a forced air cabinet to 20 percent water content, followed by rapid freezing in liquid nitrogen (-196 degree C) and rapid thawing at 45 degree C. In these conditions 29 percent of the zygotic embryos germinated in vitro. However, plantlets obtained from dehydrated zygotic embryos had stunted haustoria and lower heights. Histological analysis showed that haustorium cells were large, vacuolated, with few protein bodies. In contrast, small cells with high nucleus:cytoplasm ratio formed the shoot apical meristem of the embryos, which were the cell types with favorable characteristics for survival after exposure to liquid nitrogen. Plantlets were successfully acclimatized and showed 41+/-9 percent and 88+/-4 percent survival levels after 12 weeks of acclimatization from cryopreserved and non-cryopreserved treatments, respectively.

  19. Rab GTPases in Immunity and Inflammation

    Directory of Open Access Journals (Sweden)

    Akriti Prashar

    2017-09-01

    Full Text Available Strict spatiotemporal control of trafficking events between organelles is critical for maintaining homeostasis and directing cellular responses. This regulation is particularly important in immune cells for mounting specialized immune defenses. By controlling the formation, transport and fusion of intracellular organelles, Rab GTPases serve as master regulators of membrane trafficking. In this review, we discuss the cellular and molecular mechanisms by which Rab GTPases regulate immunity and inflammation.

  20. Rab GTPases in Immunity and Inflammation.

    Science.gov (United States)

    Prashar, Akriti; Schnettger, Laura; Bernard, Elliott M; Gutierrez, Maximiliano G

    2017-01-01

    Strict spatiotemporal control of trafficking events between organelles is critical for maintaining homeostasis and directing cellular responses. This regulation is particularly important in immune cells for mounting specialized immune defenses. By controlling the formation, transport and fusion of intracellular organelles, Rab GTPases serve as master regulators of membrane trafficking. In this review, we discuss the cellular and molecular mechanisms by which Rab GTPases regulate immunity and inflammation.

  1. Sensory profile of eleven peach cultivars

    Directory of Open Access Journals (Sweden)

    Francine Lorena Cuquel

    2012-03-01

    Full Text Available The goal of this study was to evaluate the sensory profile of eleven peach cultivars grown in an experimental orchard located in the city of Lapa (PR, Brazil in two seasons. The peach cultivars analyzed were Aurora I, Chimarrita, Chiripá, Coral, Eldorado, Granada, Leonense, Maciel, Marli, Premier, and Vanguarda. The sensory analysis was performed by previously trained panelists; 20 of them in the first season and 10 in the second season. The sensory evaluation was performed using Quantitative Descriptive Analysis, in which the following attributes were measured: appearance, aroma, flesh color, flesh firmness, flavor, and juiciness. The results showed preference for sweet, soft, and juicy fruits. Chimarrita, Chiripá, and Coral fruits showed better sensorial performance than the other peach cultivars. It was also verified that the analysis of the attributes aroma, flesh firmness, and flavor is enough for performing the sensory profile of peach fruits for in natura consumption.

  2. 21 CFR 145.170 - Canned peaches.

    Science.gov (United States)

    2010-04-01

    ... CONSUMPTION CANNED FRUITS Requirements for Specific Standardized Canned Fruits § 145.170 Canned peaches. (a... by a support through which it can freely move upward or downward. The lower end of the rod is a plane...

  3. Variability of vineyard peach tree characteristics

    Directory of Open Access Journals (Sweden)

    Zec Gordan

    2008-01-01

    Full Text Available Vineyard peach seedlings are the most important rootstock for peach in Serbia and abroad. High variability is a characteristic of the vineyard peach planting material that is used as rootstock in nursery production. Through work of many years, vineyard peach genotypes with qualitative traits were selected and collected. Seedlings that are progeny of the nine selected genotypes and resulted from self and open pollination were examined. The vineyard peach seedlings resulted from uncontrolled pollination and with different geographical origin served as reference. A goal of research was to get vineyard peach genotypes that would give more uniform generative progeny with qualitative traits. This paper presents the results of two-year research of morphological traits of more than 500 vineyard seedlings. Based on the results, positive genotypes were selected for further inbreeding. Further, the seedlings of the selection 6 have the lowest coefficients of variation for trunk thickness, tree height and number of branches, which points to the self-pollination as a good method for getting more uniform progeny. .

  4. Peach and apricot stone combustion in a bubbling fluidized bed

    Energy Technology Data Exchange (ETDEWEB)

    Kaynak, B.; Atimtay, Aysel T. [Department of Environmental Engineering, Middle East Technical University, Ankara 06531 (Turkey); Topal, H. [Department of Mechanical Engineering, Engineering and Architecture Faculty, Gazi University, Ankara 06570 (Turkey)

    2005-07-25

    peach and apricot stones are potential fuels that can be utilized for clean energy production in small-scale fruit juice industries by using BFBC.

  5. Origin and Dispersal of Domesticated Peach Palm

    Directory of Open Access Journals (Sweden)

    Charles R. Clement

    2017-11-01

    Full Text Available Peach palm (Bactris gasipaes Kunth is a Neotropical palm domesticated by Native Americans. Its domestication resulted in a set of landraces (var. gasipaes, some with very starchy fruit used for fermentation, others with an equilibrium of starch and oil used as snacks. Which of the three wild types (var. chichagui was involved and where the domestication process began are unclear, with three hypotheses under discussion: an origin in southwestern Amazonia; or in northwestern South America; or multiple origins. We reevaluate one of the wild types, defining it as the incipient domesticate, and then evaluate these hypotheses using the Brazilian peach palm Core Collection and selected herbaria samples to: (1 model the potential distributions of wild and domesticated populations; (2 identify the probable origin of domestication with a phylogeographic analysis of chloroplast DNA sequences; and (3 determine the dispersal routes after domestication using spatial analysis of genetic diversity based on 17 nuclear microsatellite loci. The two very small-fruited wild types have distinct distributions in the northern Andes region and across southern Amazonia, both under moderately humid climates, while the incipient domesticate, partly sympatric with the southern wild type, is also found along the Equatorial Andes, in a more humid climatic envelope, more similar to that of the domesticated landraces. Two distribution models for Last Glacial Maximum conditions (CCSM4, MIROC also suggest distinct distributions for the two wild populations. The chloroplast DNA phylogeographic network confirms the area of sympatry of the incipient domesticate and the southern wild type in southwestern Amazonia as the origin of domestication. The spatial patterns of genetic diversity confirm the proposal of two dispersals, one along the Ucayali River, into western Amazonia, northwestern South America and finally Central America; the other along the Madeira River into central and

  6. Regulation of ER-Golgi Transport Dynamics by GTPases in Budding Yeast

    Directory of Open Access Journals (Sweden)

    Yasuyuki Suda

    2018-01-01

    Full Text Available A large number of proteins are synthesized de novo in the endoplasmic reticulum (ER. They are transported through the Golgi apparatus and then delivered to their proper destinations. The ER and the Golgi play a central role in protein processing and sorting and show dynamic features in their forms. Ras super family small GTPases mediate the protein transport through and between these organelles. The ER-localized GTPase, Sar1, facilitates the formation of COPII transport carriers at the ER exit sites (ERES on the ER for the transport of cargo proteins from the ER to the Golgi. The Golgi-localized GTPase, Arf1, controls intra-Golgi, and Golgi-to-ER transport of cargo proteins by the formation of COPI carriers. Rab GTPases localized at the Golgi, which are responsible for fusion of membranes, are thought to establish the identities of compartments. Recent evidence suggests that these small GTPases regulate not only discrete sites for generation/fusion of transport carriers, but also membrane dynamics of the organelles where they locate to ensure the integrity of transport. Here we summarize the current understandings about the membrane traffic between these organelles and highlight the cutting-edge advances from super-resolution live imaging of budding yeast, Saccharomyces cerevisiae.

  7. Rho GTPase expression in human myeloid cells.

    Directory of Open Access Journals (Sweden)

    Suzanne F G van Helden

    Full Text Available Myeloid cells are critical for innate immunity and the initiation of adaptive immunity. Strict regulation of the adhesive and migratory behavior is essential for proper functioning of these cells. Rho GTPases are important regulators of adhesion and migration; however, it is unknown which Rho GTPases are expressed in different myeloid cells. Here, we use a qPCR-based approach to investigate Rho GTPase expression in myeloid cells.We found that the mRNAs encoding Cdc42, RhoQ, Rac1, Rac2, RhoA and RhoC are the most abundant. In addition, RhoG, RhoB, RhoF and RhoV are expressed at low levels or only in specific cell types. More differentiated cells along the monocyte-lineage display lower levels of Cdc42 and RhoV, while RhoC mRNA is more abundant. In addition, the Rho GTPase expression profile changes during dendritic cell maturation with Rac1 being upregulated and Rac2 downregulated. Finally, GM-CSF stimulation, during macrophage and osteoclast differentiation, leads to high expression of Rac2, while M-CSF induces high levels of RhoA, showing that these cytokines induce a distinct pattern. Our data uncover cell type specific modulation of the Rho GTPase expression profile in hematopoietic stem cells and in more differentiated cells of the myeloid lineage.

  8. Rab27 GTPases Distribute Extracellular Nanomaps for Invasive Growth and Metastasis: Implications for Prognosis and Treatment

    Directory of Open Access Journals (Sweden)

    Olivier De Wever

    2013-05-01

    Full Text Available The Rab27 family of small GTPases regulates exocytosis of distinct vesicle types including multivesicular endosomes, which results in the release of exosomes. Exosomes are nanometer-sized membrane vesicles that enclose soluble factors such as proteins and nucleic acids within a lipid bilayer and can travel toward distant tissues to influence multiple aspects of cell behavior. In our view that tumors are endocrine organs producing exosomes, Rab27 GTPases and their effector proteins are critical determinants for invasive growth and metastasis. Rab27 proteins and their effectors may serve as prognostic biomarkers or as targets for patient-tailored therapy.

  9. The Small Rho GTPases Rac1 and Rac2 Are Important for T-Cell Independent Antigen Responses and for Suppressing Switching to IgG2b in Mice.

    Science.gov (United States)

    Gerasimčik, Natalija; He, Minghui; Dahlberg, Carin I M; Kuznetsov, Nikolai V; Severinson, Eva; Westerberg, Lisa S

    2017-01-01

    The Rho GTPases Cdc42, Rac1, and Rac2 coordinate receptor signaling to cell adhesion, migration, and proliferation. Deletion of Rac1 and Rac2 early during B cell development leads to failure in B cell entry into the splenic white pulp. Here, we sought to understand the role of Rac1 and Rac2 in B cell functionality and during the humoral antibody response. To circumvent the migratory deficiency of B cells lacking both Rac1 and Rac2, we took the approach to inducibly delete Rac1 in Rac2 -/- B cells in the spleen (Rac1 B Rac2 -/- B cells). Rac1 B Rac2 -/- mice had normal differentiation of splenic B cell populations, except for a reduction in marginal zone B cells. Rac1 B Rac2 -/- B cells showed normal spreading response on antibody-coated layers, while both Rac2 -/- and Rac1 B Rac2 -/- B cells had reduced homotypic adhesion and decreased proliferative response when compared to wild-type B cells. Upon challenge with the T-cell-independent antigen TNP-conjugated lipopolysaccharide, Rac1 B Rac2 -/- mice showed reduced antibody response. In contrast, in response to the T-cell-dependent antigen sheep red blood cells, Rac1 B Rac2 -/- mice had increased serum titers of IgG1 and IgG2b. During in vitro Ig class switching, Rac1 B Rac2 -/- B cells had elevated germline γ2b transcripts leading to increased Ig class switching to IgG2b. Our data suggest that Rac1 and Rac2 serve an important role in regulation of the B cell humoral immune response and in suppressing Ig class switching to IgG2b.

  10. Influence of vineyard peach selections on vigour and initial yield in peach and nectarine

    Directory of Open Access Journals (Sweden)

    Zec Gordan

    2013-01-01

    Full Text Available Seedlings (progenies of eight selected vineyard peach genotypes obtained by self and open pollination were examined in this study during three consecutive years. Two progenies from unselected vineyard peach genotypes with different geographic origin were used as a standard. Peach cv. Autumn Glo and nectarine cv. Stark Red Gold were grafted on F1 generation of eight vineyard peach selections and two standard progenies. Analysis of variance indicated statistically significant differences in vigour and fruit productivity between fruit trees grafted on different progenies of vineyard peach selections. A high correlation was found between rootstock vigour and fruit production of grafted cultivars. In addition, the principal component analysis made it possible to establish similar groups of rootstocks, depending on its influence to vigour, productivity and indexes of efficiency of grafted peach and nectarine cultivar. The most promising rootstocks for those two cultivars were PSK and 7S because grafted AG and SRG have high fruit weight, initial yields and very satisfactory rootstock, scion and canopy efficiency. [Projekat Ministarstva nauke Republike Srbije, br. TR 31063

  11. Difference in photosynthetic performance among three peach ...

    African Journals Online (AJOL)

    The effects of low photosynthetic photon flux density (PPFD) on greenhouse grown peach trees ('Qingfeng': Prunus persica L. Batsch, 'NJN76': Prunus persica L. Batsch and 'Maixiang': P. persica var. nectarine) were investigated. Difference in photosynthesis rate (Pn) and stoma morphology among cultivars were studied.

  12. Consumer peach preferences and purchasing behavior: a mixed methods study.

    Science.gov (United States)

    Kelley, Kathleen M; Primrose, Rachel; Crassweller, Robert; Hayes, John E; Marini, Richard

    2016-05-01

    Peaches (Prunus persica (L.) Stokes) are grown in several regions throughout the USA, are eaten fresh, and used as ingredients in value-added processed products. An Internet survey was conducted to investigate Mid-Atlantic consumers' fresh and processed peach purchasing behaviors, and whether packaging certain numbers of peaches together, providing information about nutritional content, and other factors would increase purchases. Additionally, laboratory-based sensory testing was used to better understand peel color, texture, sweetness, sourness, and flavor preferences for cultivars commonly grown in the Mid-Atlantic region. Irrespective of fresh peach consumption frequency, certain value-added products were of interest. For some products, interest in purchasing was higher than reported purchasing behavior. Preference for certain fresh peach characteristics, such as peel color, differed between less frequent fresh peach consumers and those who consumed fresh peaches more often. Of the four peach cultivars included in the sensory test, most were liked; however, there were some cultivar differences pertaining to color, texture, sweetness, tartness, and flavor liking. Potential marketing strategies can be developed based on frequency of fresh peach consumption and household demographics. Data can be used to select peaches that best appeal to consumers. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

  13. Ecological adaptation of wild peach palm, its in situ conservation and deforestation-mediated extinction in southern Brazilian Amazonia.

    Directory of Open Access Journals (Sweden)

    Charles R Clement

    Full Text Available BACKGROUND: The Arc of Fire across southern Amazonia seasonally attracts worldwide attention as forests are cut and burned for agricultural expansion. These forests contain numerous wild relatives of native South American crops, such as peach palm. METHODOLOGY/PRINCIPAL FINDINGS: Our prospecting expeditions examined critical areas for wild peach palm in the Arc of Fire in Mato Grosso, Pará, Maranhão and Tocantins, as well as areas not previously examined in Amazonas and Amapá states. Recent digitization of the RADAM Brasil project permitted comparison among RADAM's parataxonomists' observations, previous botanical collections and our prospecting. Mapping on soils and vegetation types enabled us to hypothesize a set of ecological preferences. Wild peach palm is best adapted to Ultisols (Acrisols in open forests across the Arc of Fire and westward into the more humid western Amazonia. Populations are generally small (fewer than 10 plants on slopes above watercourses. In northern Mato Grosso and southern Pará soybean fields and pastures now occupy numerous areas where RADAM identified wild peach palm. The controversial BR-163 Highway is already eroding wild peach palm as deforestation expands. CONCLUSIONS/SIGNIFICANCE: Many of these populations are now isolated by increasing forest fragmentation, which will lead to decreased reproduction via inbreeding depression and eventual extinction even without complete deforestation. Federal conservation areas are less numerous in the Arc of Fire than in other parts of Brazilian Amazonia, although there are indigenous lands; these conservation areas contain viable populations of wild peach palm and require better protection than they are currently receiving. Ex situ conservation of these populations is not viable given the relative lack of importance of domesticated peach palm and the difficulty of maintaining even economically interesting genetic resources.

  14. Ecological adaptation of wild peach palm, its in situ conservation and deforestation-mediated extinction in southern Brazilian Amazonia.

    Science.gov (United States)

    Clement, Charles R; Santos, Ronaldo P; Desmouliere, Sylvain J M; Ferreira, Evandro J L; Neto, João Tomé Farias

    2009-01-01

    The Arc of Fire across southern Amazonia seasonally attracts worldwide attention as forests are cut and burned for agricultural expansion. These forests contain numerous wild relatives of native South American crops, such as peach palm. Our prospecting expeditions examined critical areas for wild peach palm in the Arc of Fire in Mato Grosso, Pará, Maranhão and Tocantins, as well as areas not previously examined in Amazonas and Amapá states. Recent digitization of the RADAM Brasil project permitted comparison among RADAM's parataxonomists' observations, previous botanical collections and our prospecting. Mapping on soils and vegetation types enabled us to hypothesize a set of ecological preferences. Wild peach palm is best adapted to Ultisols (Acrisols) in open forests across the Arc of Fire and westward into the more humid western Amazonia. Populations are generally small (fewer than 10 plants) on slopes above watercourses. In northern Mato Grosso and southern Pará soybean fields and pastures now occupy numerous areas where RADAM identified wild peach palm. The controversial BR-163 Highway is already eroding wild peach palm as deforestation expands. Many of these populations are now isolated by increasing forest fragmentation, which will lead to decreased reproduction via inbreeding depression and eventual extinction even without complete deforestation. Federal conservation areas are less numerous in the Arc of Fire than in other parts of Brazilian Amazonia, although there are indigenous lands; these conservation areas contain viable populations of wild peach palm and require better protection than they are currently receiving. Ex situ conservation of these populations is not viable given the relative lack of importance of domesticated peach palm and the difficulty of maintaining even economically interesting genetic resources.

  15. BAR domain proteins regulate Rho GTPase signaling.

    Science.gov (United States)

    Aspenström, Pontus

    2014-01-01

    BAR proteins comprise a heterogeneous group of multi-domain proteins with diverse biological functions. The common denominator is the Bin-Amphiphysin-Rvs (BAR) domain that not only confers targeting to lipid bilayers, but also provides scaffolding to mold lipid membranes into concave or convex surfaces. This function of BAR proteins is an important determinant in the dynamic reconstruction of membrane vesicles, as well as of the plasma membrane. Several BAR proteins function as linkers between cytoskeletal regulation and membrane dynamics. These links are provided by direct interactions between BAR proteins and actin-nucleation-promoting factors of the Wiskott-Aldrich syndrome protein family and the Diaphanous-related formins. The Rho GTPases are key factors for orchestration of this intricate interplay. This review describes how BAR proteins regulate the activity of Rho GTPases, as well as how Rho GTPases regulate the function of BAR proteins. This mutual collaboration is a central factor in the regulation of vital cellular processes, such as cell migration, cytokinesis, intracellular transport, endocytosis, and exocytosis.

  16. Regulation of anthocyanin biosynthesis in peach fruits.

    Science.gov (United States)

    Rahim, Md Abdur; Busatto, Nicola; Trainotti, Livio

    2014-11-01

    MYB10.1 and MYB10.3, with bHLH3, are the likely regulators of anthocyanin biosynthesis in peach fruit. MYB10.1/2/3 forms a cluster on the same genomic fragment where the Anther color ( Ag ) trait is located. Anthocyanins are bioactive compounds responsible for the pigmentation of many plant parts such as leaves, flowers, fruits and roots, and have potential benefits to human health. In peach [Prunus persica (L.) Batsch], peel color is a key determinant for fruit quality and is regulated by flavonoids including anthocyanins. The R2R3 MYB transcription factors (TFs) control the expression of anthocyanin biosynthetic genes with the help of co-activators belonging to the basic-helix-loop-helix (bHLH) and WD40 repeat families. In the peach genome six MYB10-like and three bHLH-like TFs were identified as candidates to be the regulators of the anthocyanin accumulation, which, in yellow flesh fruits, is highest in the peel, abundant in the part of the mesocarp surrounding the stone and lowest in the mesocarp. The expression of MYB10.1 and MYB10.3 correlates with anthocyanin levels of different peach parts. They also have positive correlation with the expression of key structural genes of the anthocyanin pathway, such as CHS, F3H, and UFGT. Functions of peach MYB10s were tested in tobacco and shown to activate key genes in the anthocyanin pathway when bHLHs were co-expressed as partners. Overexpression of MYB10.1/bHLH3 and MYB10.3/bHLH3 activated anthocyanin production by up-regulating NtCHS, NtDFR and NtUFGT while other combinations were not, or much less, effective. As three MYB10 genes are localized in a genomic region where the Ag trait, responsible for anther pigmentation, is localized, it is proposed they are key determinant to introduce new peach cultivars with higher antioxidant level and pigmented fruit.

  17. Fungicide efficacy in peach rusty spot control in Serbia

    OpenAIRE

    Dolovac Nenad; Miletić Novica; Aleksić Goran; Savić Dušan; Živković Svetlana; Trkulja Nenad; Bulajić Aleksandra

    2010-01-01

    Rusty Spot has long been known as a harmful peach disease in many parts of the world. During the past several years, rusty spot infection of the late-maturing peach cultivars (Summerset, Suncrest, Fayette and O’Henry) caused significant yield losses in Serbia. Although the etiology of the disease is still unknown, there are numerous studies attempting to set a strategy for its control and recommend appropriate chemical and other peach protection methods. Ho...

  18. Transcriptomic and Functional Analyses Reveal That PpGLK1 Regulates Chloroplast Development in Peach (Prunus persica

    Directory of Open Access Journals (Sweden)

    Min Chen

    2018-01-01

    Full Text Available Peach is an ideal species for fruit tree research because of its small, fully sequenced genome. Chloroplast development is dependent on the tight cooperation between the nuclear and plastid genomes, and is regulated by GLK transcription factors. In this work, the pigment content was monitored and the chloroplast-to-chromoplast conversion during the fruit ripening was visualized by transmission electron microscopy. Localization and expression analyses showed that PpGLK1 was located in the nucleus and expressed mainly in the leaves and fruit skin. A transcriptome analysis showed that PpGLK1 and its target genes were significantly differentially expressed in ripening peach fruit skin. PpGLK1 silencing affected chlorophyll accumulation in peach leaves and fruits. Overexpression of PpGLK1 rescued the phenotypes of the Arabidopsis Atglk1Atglk2 double mutant and the tomato uniform ripening mutant. The results of a yeast two-hybrid analysis showed that PpGLK1 is autoactivated and that PpGLK1 (301-542 a.a. interacted with PpARF5. Together, our results indicate that PpGLK1 regulates chloroplast development in green tissues in peach. Therefore, it may be a promising target gene for improving the production and quality of peach by genetic engineering and breeding approaches.

  19. Evolutionary Genomics of Peach and Almond Domestication

    Directory of Open Access Journals (Sweden)

    Dianne Velasco

    2016-12-01

    Full Text Available The domesticated almond [Prunus dulcis (L. Batsch] and peach [P. persica (Mill. D. A. Webb] originated on opposite sides of Asia and were independently domesticated ∼5000 yr ago. While interfertile, they possess alternate mating systems and differ in a number of morphological and physiological traits. Here, we evaluated patterns of genome-wide diversity in both almond and peach to better understand the impacts of mating system, adaptation, and domestication on the evolution of these taxa. Almond has around seven times the genetic diversity of peach, and high genome-wide FST values support their status as separate species. We estimated a divergence time of ∼8 MYA (million years ago, coinciding with an active period of uplift in the northeast Tibetan Plateau and subsequent Asian climate change. We see no evidence of a bottleneck during domestication of either species, but identify a number of regions showing signatures of selection during domestication and a significant overlap in candidate regions between peach and almond. While we expected gene expression in fruit to overlap with candidate selected regions, instead we find enrichment for loci highly differentiated between the species, consistent with recent fossil evidence suggesting fruit divergence long preceded domestication. Taken together, this study tells us how closely related tree species evolve and are domesticated, the impact of these events on their genomes, and the utility of genomic information for long-lived species. Further exploration of this data will contribute to the genetic knowledge of these species and provide information regarding targets of selection for breeding application, and further the understanding of evolution in these species.

  20. Evolutionary Genomics of Peach and Almond Domestication.

    Science.gov (United States)

    Velasco, Dianne; Hough, Josh; Aradhya, Mallikarjuna; Ross-Ibarra, Jeffrey

    2016-12-07

    The domesticated almond [Prunus dulcis (L.) Batsch] and peach [P. persica (Mill.) D. A. Webb] originated on opposite sides of Asia and were independently domesticated ∼5000 yr ago. While interfertile, they possess alternate mating systems and differ in a number of morphological and physiological traits. Here, we evaluated patterns of genome-wide diversity in both almond and peach to better understand the impacts of mating system, adaptation, and domestication on the evolution of these taxa. Almond has around seven times the genetic diversity of peach, and high genome-wide [Formula: see text] values support their status as separate species. We estimated a divergence time of ∼8 MYA (million years ago), coinciding with an active period of uplift in the northeast Tibetan Plateau and subsequent Asian climate change. We see no evidence of a bottleneck during domestication of either species, but identify a number of regions showing signatures of selection during domestication and a significant overlap in candidate regions between peach and almond. While we expected gene expression in fruit to overlap with candidate selected regions, instead we find enrichment for loci highly differentiated between the species, consistent with recent fossil evidence suggesting fruit divergence long preceded domestication. Taken together, this study tells us how closely related tree species evolve and are domesticated, the impact of these events on their genomes, and the utility of genomic information for long-lived species. Further exploration of this data will contribute to the genetic knowledge of these species and provide information regarding targets of selection for breeding application, and further the understanding of evolution in these species. Copyright © 2016 Velasco et al.

  1. Evolutionary Genomics of Peach and Almond Domestication

    Science.gov (United States)

    Velasco, Dianne; Hough, Josh; Aradhya, Mallikarjuna; Ross-Ibarra, Jeffrey

    2016-01-01

    The domesticated almond [Prunus dulcis (L.) Batsch] and peach [P. persica (Mill.) D. A. Webb] originated on opposite sides of Asia and were independently domesticated ∼5000 yr ago. While interfertile, they possess alternate mating systems and differ in a number of morphological and physiological traits. Here, we evaluated patterns of genome-wide diversity in both almond and peach to better understand the impacts of mating system, adaptation, and domestication on the evolution of these taxa. Almond has around seven times the genetic diversity of peach, and high genome-wide FST values support their status as separate species. We estimated a divergence time of ∼8 MYA (million years ago), coinciding with an active period of uplift in the northeast Tibetan Plateau and subsequent Asian climate change. We see no evidence of a bottleneck during domestication of either species, but identify a number of regions showing signatures of selection during domestication and a significant overlap in candidate regions between peach and almond. While we expected gene expression in fruit to overlap with candidate selected regions, instead we find enrichment for loci highly differentiated between the species, consistent with recent fossil evidence suggesting fruit divergence long preceded domestication. Taken together, this study tells us how closely related tree species evolve and are domesticated, the impact of these events on their genomes, and the utility of genomic information for long-lived species. Further exploration of this data will contribute to the genetic knowledge of these species and provide information regarding targets of selection for breeding application, and further the understanding of evolution in these species. PMID:27707802

  2. Role of Arf GTPases in fungal morphogenesis and virulence.

    Directory of Open Access Journals (Sweden)

    Hayet Labbaoui

    2017-02-01

    Full Text Available Virulence of the human fungal pathogen Candida albicans depends on the switch from budding to filamentous growth, which requires sustained membrane traffic and polarized growth. In many organisms, small GTPases of the Arf (ADP-ribosylation factor family regulate membrane/protein trafficking, yet little is known about their role in fungal filamentous growth. To investigate these GTPases in C. albicans, we generated loss of function mutants in all 3 Arf proteins, Arf1-Arf3, and 2 Arf-like proteins, Arl1 and Arl3. Our results indicate that of these proteins, Arf2 is required for viability and sensitivity to antifungal drugs. Repressible ARF2 expression results in defects in filamentous growth, cell wall integrity and virulence, likely due to alteration of the Golgi. Arl1 is also required for invasive filamentous growth and, although arl1/arl1 cells can initiate hyphal growth, hyphae are substantially shorter than that of the wild-type, due to the inability of this mutant to maintain hyphal growth at a single site. We show that this defect does not result from an alteration of phospholipid distribution and is unlikely to result from the sole Golgin Imh1 mislocalization, as Imh1 is not required for invasive filamentous growth. Rather, our results suggest that the arl1/arl1 hyphal growth defect results from increased secretion in this mutant. Strikingly, the arl1/arl1 mutant is drastically reduced in virulence during oropharyngeal candidiasis. Together, our results highlight the importance of Arl1 and Arf2 as key regulators of hyphal growth and virulence in C. albicans and identify a unique function of Arl1 in secretion.

  3. Individual sugar and acid composition within southeastern peach germplasm

    Science.gov (United States)

    eaches grown in the southeast are valued for their acidic, sweet flavor. A complex mixture of various sugars and acids at different ratios play a key role in determining these unique peach flavor attributes. To understand the flavor profile of fresh market peaches, individual sugar and acid compone...

  4. ATPase and GTPase Tangos Drive Intracellular Protein Transport.

    Science.gov (United States)

    Shan, Shu-Ou

    2016-12-01

    The GTPase superfamily of proteins provides molecular switches to regulate numerous cellular processes. The 'GTPase switch' paradigm, in which external regulatory factors control the switch of a GTPase between 'on' and 'off' states, has been used to interpret the regulatory mechanism of many GTPases. However, recent work unveiled a class of nucleotide hydrolases that do not adhere to this classical paradigm. Instead, they use nucleotide-dependent dimerization cycles to regulate key cellular processes. In this review article, recent studies of dimeric GTPases and ATPases involved in intracellular protein targeting are summarized. It is suggested that these proteins can use the conformational plasticity at their dimer interface to generate multiple points of regulation, thereby providing the driving force and spatiotemporal coordination of complex cellular pathways. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Regulation of cerebral cortex development by Rho GTPases: insights from in vivo studies

    Directory of Open Access Journals (Sweden)

    Roberta eAzzarelli

    2015-01-01

    Full Text Available The cerebral cortex is the site of higher human cognitive and motor functions. Histologically, it is organized into six horizontal layers, each containing unique populations of molecularly and functionally distinct excitatory projection neurons and inhibitory interneurons. The stereotyped cellular distribution of cortical neurons is crucial for the formation of functional neural circuits and it is predominantly established during embryonic development. Cortical neuron development is a multiphasic process characterized by sequential steps of neural progenitor proliferation, cell cycle exit, neuroblast migration and neuronal differentiation. This series of events requires an extensive and dynamic remodeling of the cell cytoskeleton at each step of the process. As major regulators of the cytoskeleton, the family of small Rho GTPases has been shown to play essential functions in cerebral cortex development. Here we review in vivo findings that support the contribution of Rho GTPases to cortical projection neuron development and we address their involvement in the etiology of cerebral cortex malformations.

  6. Micropropagation of peach rootstocks and cultivars.

    Science.gov (United States)

    Balla, Ildikó; Mansvelt, Lucienne

    2013-01-01

    Peach (Prunus persica (L.) Batsch) is one of the most popular stone fruits, commercially produced largely in Mediterranean and, to a lesser extent, in continental climatic conditions. Several breeding programs with different aims release annually large numbers of new cultivars. Micropropagation offers a suitable method to provide the growers of sufficient quantities of rootstocks, as well as of pathogen-free planting material of old and new cultivars.An effective four-step micropropagation procedure for cultivar and rootstock production is described here, based on the use of modified MS and WPM media. The health status of the initial shoot tips is very important, also because the growth and proliferation rate of shoot cultures from virus-infected clones are generally very poor. Proliferation and elongation phases depend on the major macro-elements, as well as the content and ratio of plant growth regulators. It is important to grow the cultures at 22°C, as hyperhydricity may develop at higher temperatures. Although sucrose is the most common carbon source used during proliferation and rooting, for some peach cultivars and rootstocks the replacement of sucrose (10 g/L) with glucose (20 g/L) in the rooting medium improves the rooting and survival rates of plants in the acclimatization phase. The rooting rate of the rootstock "Cadaman" is improved with the chelated form of iron FeEDDHA at 150 mg/L. Rooted plants are acclimatized in greenhouse under high humidity conditions.

  7. Prunus hybrids rootstocks for flat peach

    Directory of Open Access Journals (Sweden)

    Pilar Legua

    2012-02-01

    Full Text Available Peach (Prunus persica L. is the most important stone fruit tree grown in Spain and is the second most important fruit crop in Europe. The influence of eight Prunus rootstocks (GF-677, Krymsk® 86, PADAC 97-36, PADAC 99-05, PADAC 9912-03, PADAC 0024-01, PAC 0021-01 and PAC 0022-01 on vigor, yield and fruit quality traits of 'UFO 3' flat peach cultivar was studied. The highest trunk cross sectional area was exhibited by GF-677 and the lowest by PADAC 99-05, while intermediate values were found on the other rootstocks. The highest yield efficiency was found on PADAC 99-05, PAC 0021-01, PAC 0022-01 and PADAC 0024-01 and the lowest was shown on Krymsk® 86. The fruit quality parameters measured were color, fruit and stone weights, equatorial diameter, pulp thickness, pulp yield, firmness, pH, soluble solids content and titratable acidity. 'UFO 3' grafted on GF-677 resulted in the largest fruit weight, while the smallest was on PADAC 99-05. Fruits of 'UFO 3' showed a tendency to have higher firmness, higher red colored skin and RI when grafted on PADAC 99-05.

  8. Integrins and small GTPases as modulators of phagocytosis.

    Science.gov (United States)

    Sayedyahossein, Samar; Dagnino, Lina

    2013-01-01

    Phagocytosis is the mechanism whereby cells engulf large particles. This process has long been recognized as a critical component of the innate immune response, which constitutes the organism's defense against microorganisms. In addition, phagocytic internalization of apoptotic cells or cell fragments plays important roles in tissue homeostasis and remodeling. Phagocytosis requires target interactions with receptors on the plasma membrane of the phagocytic cell. Integrins have been identified as important mediators of particle clearance, in addition to their well-established roles in cell adhesion, migration and mechanotransduction. Indeed, these ubiquitously expressed proteins impart phagocytic capacity to epithelial, endothelial and mesenchymal cell types. The importance of integrins in particle internalization is emphasized by the ability of microbial and viral pathogens to exploit their signaling pathways to invade host cells, and by the wide variety of disorders that arise from abnormalities in integrin-dependent phagocytic uptake. Copyright © 2013 Elsevier Inc. All rights reserved.

  9. Characterization of a Rab11-like GTPase, EhRab11, of Entamoeba histolytica.

    Science.gov (United States)

    McGugan, Glen C; Temesvari, Lesly A

    2003-07-01

    The Entamoeba histolytica Rab11 family of small molecular weight GTPases consists of three members, EhRab11, EhRab11B, and EhRab11C. The functions of these Rabs in Entamoeba have not been determined. Therefore, as an approach to elucidate the role of the Rab11 family of GTPases in Entamoeba, immunofluorescence microscopy was undertaken to define the subcellular localization of one member of this family, EhRab11. Under conditions of growth, EhRab11 displayed a punctate pattern in the cytoplasm of trophozoites. EhRab11 did not colocalize with markers for the Golgi apparatus, endoplasmic reticulum, pinosomes, phagosomes, or compartments formed by receptor-mediated endocytosis, suggesting that this Rab may not play a role in vesicle trafficking between these organelles. Under conditions of iron and serum starvation, EhRab11 was translocated to the periphery of the cell. The altered cellular localization was accompanied by multinucleation of the cells as well as the acquisition of detergent resistance by the cells, features that are characteristic of Entamoeba cysts. The translocation of EhRab11 to the periphery of the cell during iron and serum starvation was specific as the subcellular localizations of two other Rab GTPases, EhRab7 and EhRabA, were not altered under the same conditions. In addition, the formation of multinucleated cells by inhibition of cytokinesis was not sufficient to induce the translocation of EhRab11 to the cell periphery. Taken together, the data suggest that iron and serum starvation may induce encystation in E. histolytica and that EhRab11 may play a role in this process. Moreover, these studies are the first to describe a putative role for a Rab GTPase in encystation in Entamoeba sp.

  10. Reverse engineering GTPase programming languages with reconstituted signaling networks.

    Science.gov (United States)

    Coyle, Scott M

    2016-07-02

    The Ras superfamily GTPases represent one of the most prolific signaling currencies used in Eukaryotes. With these remarkable molecules, evolution has built GTPase networks that control diverse cellular processes such as growth, morphology, motility and trafficking. (1-4) Our knowledge of the individual players that underlie the function of these networks is deep; decades of biochemical and structural data has provided a mechanistic understanding of the molecules that turn GTPases ON and OFF, as well as how those GTPase states signal by controlling the assembly of downstream effectors. However, we know less about how these different activities work together as a system to specify complex dynamic signaling outcomes. Decoding this molecular "programming language" would help us understand how different species and cell types have used the same GTPase machinery in different ways to accomplish different tasks, and would also provide new insights as to how mutations to these networks can cause disease. We recently developed a bead-based microscopy assay to watch reconstituted H-Ras signaling systems at work under arbitrary configurations of regulators and effectors. (5) Here we highlight key observations and insights from this study and propose extensions to our method to further study this and other GTPase signaling systems.

  11. Study of tritium permeation through Peach Bottom Steam Generator tubes

    International Nuclear Information System (INIS)

    Yang, L.; Baugh, W.A.; Baldwin, N.L.

    1977-06-01

    The report describes the equipment developed, samples tested, procedures used, and results obtained in the tritium permeation tests conducted on steam generator tubing samples which were removed from the Peach Bottom Unit No. 1 reactor

  12. Inter-cellular transport of ran GTPase.

    Directory of Open Access Journals (Sweden)

    Deepak Khuperkar

    Full Text Available Ran, a member of the Ras-GTPase superfamily, has a well-established role in regulating the transport of macromolecules across the nuclear envelope (NE. Ran has also been implicated in mitosis, cell cycle progression, and NE formation. Over-expression of Ran is associated with various cancers, although the molecular mechanism underlying this phenomenon is unclear. Serendipitously, we found that Ran possesses the ability to move from cell-to-cell when transiently expressed in mammalian cells. Moreover, we show that the inter-cellular transport of Ran is GTP-dependent. Importantly, Ran displays a similar distribution pattern in the recipient cells as that in the donor cell and co-localizes with the Ran binding protein Nup358 (also called RanBP2. Interestingly, leptomycin B, an inhibitor of CRM1-mediated export, or siRNA mediated depletion of CRM1, significantly impaired the inter-cellular transport of Ran, suggesting a function for CRM1 in this process. These novel findings indicate a possible role for Ran beyond nucleo-cytoplasmic transport, with potential implications in inter-cellular communication and cancers.

  13. Station blackout calculations for Peach Bottom

    International Nuclear Information System (INIS)

    Hodge, S.A.

    1985-01-01

    A calculational procedure for the Station Blackout Severe Accident Sequence at Browns Ferry Unit One has been repeated with plant-specific application to one of the Peach Bottom Units. The only changes required in code input are with regard to the primary continment concrete, the existence of sprays in the secondary containment, and the size of the refueling bay. Combustible gas mole fractions in the secondary containment of each plant during the accident sequence are determined. It is demonstrated why the current state-of-the-art corium/concrete interaction code is inadequate for application to the study of Severe Accident Sequences in plants with the BWR MK I or MK II containment design

  14. Analysis of Peach Bottom turbine trip tests

    International Nuclear Information System (INIS)

    Cheng, H.S.; Lu, M.S.; Hsu, C.J.; Shier, W.G.; Diamond, D.J.; Levine, M.M.; Odar, F.

    1979-01-01

    Current interest in the analysis of turbine trip transients has been generated by the recent tests performed at the Peach Bottom (Unit 2) reactor. Three tests, simulating turbine trip transients, were performed at different initial power and coolant flow conditions. The data from these tests provide considerable information to aid qualification of computer codes that are currently used in BWR design analysis. The results are presented of an analysis of a turbine trip transient using the RELAP-3B and the BNL-TWIGL computer codes. Specific results are provided comparing the calculated reactor power and system pressures with the test data. Excellent agreement for all three test transients is evident from the comparisons

  15. ADVANCES IN PEACH, NECTARINE AND PLUM PROPAGATION

    Directory of Open Access Journals (Sweden)

    NEWTON ALEX MAYER

    2017-12-01

    Full Text Available ABSTRACT Nursery trees of stone fruits (Prunus spp. are traditionally produced by union of two distinct genotypes - the rootstock and the scion - which, by grafting, form a composite plant that will be maintained throughout of all plant life. In Brazil, the rootstocks are predominantly seed propagated and therefore usually results in heterogeneous trees for vigor and edaphic adaptation. However, with advances in rootstock breeding programs that released cultivars and certification in several countries (notably in Europe, the system will come gradually evolving for vegetative propagation (cuttings and tissue culture and use of seeds of selected rootstocks with specific characteristics and potted nursery trees production. For scion cultivar propagation, the budding system (with its many variations has predominantly been adopted in major producing countries. This review had as objective to comment main propagation methods adopted for rootstocks and scion in peach, nectarine and plum, and recent technical progress obtained as well as the needs of improvement for nursery tree production.

  16. Atmospheric fluoride levels in some Ontario peach orchards

    Energy Technology Data Exchange (ETDEWEB)

    Drowley, W B; Rayner, A C; Jephcott, C M

    1963-10-01

    As a result of suture injury to peaches, a survey was taken to determine monthly atmospheric fluoride levels in the soft fruit-growing area of the Niagara Peninsula of Ontario. Although the injury was confined to a few orchards, it was found necessary to locate stations for fluoride sampling over a wide area. The limed-paper candle method was used for fluoride collection. Unwashed and washed peach leaves were analyzed for fluoride content. Fluoride was separated from all samples by the Willard-Winter distillation procedure and estimated colorimetrically. It was found that a general background level of atmospheric fluoride existed in the whole surveyed area and that the levels were highest in and near the area of peach injury. The fluoride content of peach leaves was also highest in this area. The coincidence of high fluoride levels and suture injury to peaches suggests that atmospheric fluoride was the causal agent. When lime sprays were applied to peach trees in the damaged area, a reduction in the incidence of suture injury was observed.

  17. Feeding behavior of lambs fed diets containing peach palm meal

    Directory of Open Access Journals (Sweden)

    Alana Batista dos Santos

    2016-06-01

    Full Text Available The aim of this study was to evaluate the feeding behavior of lambs fed diets containing peach palm meal replacing maize (0, 10, 40, 60, and 85% DM. Thirty Santa Inês sheep with an average initial body weight of 21.6 ± 0.87 kg were distributed in a completely randomized design with five diets and six replicates. Feeding time in min kg–1 DM and min kg–1 NDFap increased by 34 min and 99.6 min, respectively, with each level of substitution of maize for the peach palm meal. Rumination and chewing times, in min kg–1 DM and min kg–1 NDF, also increased in response to the substitution of maize for peach palm meal. When expressed in min day–1, rumination and chewing activities decreased by 12.4 and 14.6 min, respectively, as the amount of peach palm meal in the concentrate was increased. The time spent idle increased linearly (P < 0.05, by 14.6 min day–1, with the replacement levels, compared with the control diet. Peach palm meal in the composition of sheep diets reduces the intakes of dry matter and fiber and decreases the feed and rumination efficiencies. Replacing maize by peach palm meal increases the feeding time and rumination and chewing activities of feedlot lambs.

  18. Peach Bottom HTGR decommissioning and component removal

    International Nuclear Information System (INIS)

    Kohler, E.J.; Steward, K.P.; Iacono, J.V.

    1977-07-01

    The prime objective of the Peach Bottom End-of-Life Program was to validate specific HTGR design codes and predictions by comparison of actual and predicted physics, thermal, fission product, and materials behavior in Peach Bottom. Three consecutive phases of the program provide input to the HTGR design methods verifications: (1) Nondestructive fuel and circuit gamma scanning; (2) removal of steam generator and primary circuit components; and (3) Laboratory examinations of removed components. Component removal site work commenced with establishment of restricted access areas and installation of controlled atmosphere tents to retain relative humidity at <30%. A mock-up room was established to test and develop the tooling and to train operators under simulated working conditions. Primary circuit ducting samples were removed by trepanning, and steam generator access was achieved by a combination of arc gouging and grinding. Tubing samples were removed using internal cutters and external grinding. Throughout the component removal phase, strict health physics, safety, and quality assurance programs were implemented. A total of 148 samples of primary circuit ducting and steam generator tubing were removed with no significant health physics or safety incidents. Additionally, component removal served to provide access fordetermination of cesium plateout distribution by gamma scanning inside the ducts and for macroexamination of the steam generator from both the water and helium sides. Evaluations are continuing and indicate excellent performance of the steam generator and other materials, together with close correlation of observed and predicted fission product plateout distributions. It is concluded that such a program of end-of-life research, when appropriately coordinated with decommissioning activities, can significantly advance nuclear plant and fuel technology development

  19. Population survey of phytoseiid mites and spider mites on peach leaves and wild plants in Japanese peach orchard.

    Science.gov (United States)

    Wari, David; Yamashita, Jun; Kataoka, Yoko; Kohara, Yoko; Hinomoto, Norihide; Kishimoto, Hidenari; Toyoshima, Shingo; Sonoda, Shoji

    2014-07-01

    A population survey of phytoseiid mites and spider mites was conducted on peach leaves and wild plants in Japanese peach orchards having different pesticide practices. The phytoseiid mite species composition on peach leaves and wild plants, as estimated using quantitative sequencing, changed during the survey period. Moreover, it varied among study sites. The phytoseiid mite species compositions were similar between peach leaves and some wild plants, such as Veronica persica, Paederia foetida, Persicaria longiseta, and Oxalis corniculata with larger quantities of phytoseiid mites, especially after mid-summer. A PCR-based method to detect the ribosomal ITS sequences of Tetranychus kanzawai and Panonychus mori from phytoseiid mites was developed. Results showed that Euseius sojaensis (specialized pollen feeder/generalist predator) uses both spider mites as prey in the field.

  20. Is home-based palliative care cost-effective? An economic evaluation of the Palliative Care Extended Packages at Home (PEACH) pilot.

    Science.gov (United States)

    McCaffrey, Nikki; Agar, Meera; Harlum, Janeane; Karnon, Jonathon; Currow, David; Eckermann, Simon

    2013-12-01

    The aim of this study was to evaluate the cost-effectiveness of a home-based palliative care model relative to usual care in expediting discharge or enabling patients to remain at home. Economic evaluation of a pilot randomised controlled trial with 28 days follow-up. Mean costs and effectiveness were calculated for the Palliative Care Extended Packages at Home (PEACH) and usual care arms including: days at home; place of death; PEACH intervention costs; specialist palliative care service use; acute hospital and palliative care unit inpatient stays; and outpatient visits. PEACH mean intervention costs per patient ($3489) were largely offset by lower mean inpatient care costs ($2450) and in this arm, participants were at home for one additional day on average. Consequently, PEACH is cost-effective relative to usual care when the threshold value for one extra day at home exceeds $1068, or $2547 if only within-study days of hospital admission are costed. All estimates are high uncertainty. The results of this small pilot study point to the potential of PEACH as a cost-effective end-of-life care model relative to usual care. Findings support the feasibility of conducting a definitive, fully powered study with longer follow-up and comprehensive economic evaluation.

  1. Co-combustion of peach and apricot stone with coal in a bubbling fluidized bed

    Energy Technology Data Exchange (ETDEWEB)

    Atimtay, Aysel T.; Kaynak, Burcak [Department of Environmental Engineering, Middle East Technical University, Ankara 06531 (Turkey)

    2008-02-15

    In this study a bubbling fluidized bed combustor (BFBC) having an inside diameter of 102 mm and a height of 900 mm was used to investigate the co-combustion characteristics of peach and apricot stones produced as a waste from the fruit juice industry with coal. A lignite coal was used for co-combustion. On-line concentrations of O{sub 2}, CO, CO{sub 2}, SO{sub 2}, NO{sub X} and total hydrocarbons (C{sub m}H{sub n}) were measured in the flue gas during combustion experiments. Variations of emissions of various pollutants were studied by changing the operating parameters (excess air ratio, fluidization velocity, and fuel feed rate). Temperature distribution along the bed was measured with thermocouples. For co-combustion of apricot and peach fruit stones with a lignite coal, various ratios of biomass to coal ranging from 0 to 100 wt.% were tested. For the peach stone co-combustion tests, efficiencies are about 98% and for the apricot stone co-combustion tests, efficiencies ranged between 94.7% and 96.9% for 25%, 50% and 75% of apricot stone in the fuel mixture. The results of this study have shown that as the biomass ratio in the fuel mixture increases, the combustion takes place at the upper regions of the main column. This causes higher temperatures in the freeboard than the bed. Also the CO and hydrocarbon (C{sub m}H{sub n}) emissions increase as the biomass percentage increases in the fuel mixture. This causes decrease in the combustion efficiency. These results suggest that peach and apricot stones are potential fuels that can be utilized for clean energy production in small-scale fruit juice industries by using BFBC. The percentage of peach stones or apricot stones in the fuel mixture is suggested to be below 50 wt.% in order to obtain the emission limits of EU. During the design of the BFBC, one has to be careful about the volatile matter (VM) content of the biomass. For the complete combustion of the VM, longer freeboard or secondary air addition should be

  2. Spatial and temporal variation of light inside peach trees

    International Nuclear Information System (INIS)

    Genard, M.; Baret, F.

    1994-01-01

    Gap fractions measured with hemispherical photographs were used to describe spatial and temporal variations of diffuse and direct light fractions transmitted to shoots within peach trees. For both cultivars studied, spatial variability of daily diffuse and direct light transmitted to shoots was very high within the tree. Diffuse and daily direct light fractions transmitted to shoots increased with shoot height within the tree and for more erect shoots. Temporal variations of hourly direct light were also large among shoots. Hourly direct light fractions transmitted to shoots were analyzed using recent developments in multivariate exploratory analysis. A gradient was observed between shoots sunlit almost all day and other shoots almost never sunlit. Well sunlit shoots were mostly located at the top of the tree and were more erect. Shoots located in the outer parts of the tree crown were slightly but significantly more sunlit than others for one cultivar. Principal component analysis additionally discriminated shoots according to the time of the day they were sunlit. This classification was related to shoot compass position for one cultivar. Spatial location of the shoot in the tree explained only a small part of light climate variability. Consequences of modeling light climate within the tree are discussed

  3. Guanine nucleotide exchange factor αPIX leads to activation of the Rac 1 GTPase/glycogen phosphorylase pathway in interleukin (IL)-2-stimulated T cells

    DEFF Research Database (Denmark)

    Llavero, Francisco; Urzelai, Bakarne; Osinalde, Nerea

    2015-01-01

    Recently, we have reported that the active form of Rac 1 GTPase binds to the glycogen phosphorylase muscle isoform (PYGM) and modulates its enzymatic activity leading to T cell proliferation. In the lymphoid system, Rac 1 and in general other small GTPases of the Rho family participate...... in the signaling cascades that are activated after engagement of the T cell antigen receptor. However, little is known about the IL-2-dependent Rac 1 activator molecules. For the first time, a signaling pathway leading to the activation of Rac 1/PYGM in response to IL-2-stimulated T cell proliferation is described....... More specifically, αPIX, a known guanine nucleotide exchange factor for the small GTPases of the Rho family, preferentially Rac 1, mediates PYGM activation in Kit 225 T cells stimulated with IL-2. Using directed mutagenesis, phosphorylation of αPIX Rho-GEF serines 225 and 488 is required for activation...

  4. Morphological and genetic perspectives of peach fruit responses to spring frost

    Science.gov (United States)

    Spring frost is one of the most unpredictable cropping factors in many peach production areas. A severe spring frost can wipe out an entire peach crop whereas a mild spring frost may naturally help thinning as is common practice in commercial peach production. The extent of frost damage depends on t...

  5. PeachVar-DB: A Curated Collection of Genetic Variations for the Interactive Analysis of Peach Genome Data.

    Science.gov (United States)

    Cirilli, Marco; Flati, Tiziano; Gioiosa, Silvia; Tagliaferri, Ilario; Ciacciulli, Angelo; Gao, Zhongshan; Gattolin, Stefano; Geuna, Filippo; Maggi, Francesco; Bottoni, Paolo; Rossini, Laura; Bassi, Daniele; Castrignanò, Tiziana; Chillemi, Giovanni

    2018-01-01

    Applying next-generation sequencing (NGS) technologies to species of agricultural interest has the potential to accelerate the understanding and exploration of genetic resources. The storage, availability and maintenance of huge quantities of NGS-generated data remains a major challenge. The PeachVar-DB portal, available at http://hpc-bioinformatics.cineca.it/peach, is an open-source catalog of genetic variants present in peach (Prunus persica L. Batsch) and wild-related species of Prunus genera, annotated from 146 samples publicly released on the Sequence Read Archive (SRA). We designed a user-friendly web-based interface of the database, providing search tools to retrieve single nucleotide polymorphism (SNP) and InDel variants, along with useful statistics and information. PeachVar-DB results are linked to the Genome Database for Rosaceae (GDR) and the Phytozome database to allow easy access to other external useful plant-oriented resources. In order to extend the genetic diversity covered by the PeachVar-DB further, and to allow increasingly powerful comparative analysis, we will progressively integrate newly released data. © The Author 2017. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  6. Fungicide Efficacy in Peach Rusty Spot Control in Serbia

    Directory of Open Access Journals (Sweden)

    Nenad Dolovac

    2010-01-01

    Full Text Available Rusty Spot has long been known as a harmful peach disease in many parts of the world. During the past several years, rusty spot infection of the late-maturing peach cultivars (Summerset, Suncrest, Fayette and O’Henry caused significant yield losses in Serbia.Although the etiology of the disease is still unknown, there are numerous studies attempting to set a strategy for its control and recommend appropriate chemical and other peach protection methods. However, because of specific environmental conditions in Serbia, recommended protection method using repeated fungicide treatments, starting from petal fall, did not prove to be efficient and the rate of infection in some susceptible peach cultivars reached 100%. In 2003 and 2004 a field trial was conducted in order to test the efficacy of fungicides (a.i. kresoxim-methyl, flusilazole and sulfur for the efficient control of Rusty Spot epidemics. The trial was carried out under conditions of natural infection on the peach cv. Summerset at the locality of Bela Crkva, Serbia. In the ntreated control plots,high disease incidence was recorded with the percentage of affected fruit surface ranging from 33.5% in the first, up to the 35.4% in the second year of the trail. Among fungicides included in the trial, kresoxim-methyl proved to be the most efficient (90.25% in the first and 91.12% in the second year of the trial, flusilazole exhibited lower efficacy (87.28% and 80.61%, respectively while sulfur was the least efficient 82.33% and 80.30%, respectively. Determination of the most efficient fungicide for the peach rusty spot control in Serbia provides basic nformation for further investigations which will include optimization of treatment terms, as well as additional agro-technical control measures.

  7. Apple and peach consumption habits across European countries.

    Science.gov (United States)

    Konopacka, D; Jesionkowska, K; Kruczyńska, D; Stehr, R; Schoorl, F; Buehler, A; Egger, S; Codarin, S; Hilaire, C; Höller, I; Guerra, W; Liverani, A; Donati, F; Sansavini, S; Martinelli, A; Petiot, C; Carbó, J; Echeverria, G; Iglesias, I; Bonany, J

    2010-12-01

    The aim of this study was to gain information concerning apple and peach consumption frequency within different European countries in relation to age and gender. The survey was a part of a complex experiment with the aim of evaluating consumers' preferences towards new varieties, and the data is based on the self-reported declarations of respondents, male and female, between 15 and 70 years old. 4271 consumers from 7 European countries were invited to supply information about their apple consumption habits, whereas 499 respondents from 5 countries answered questions relating to frequency of peach and nectarine consumption. In both, the apple and the peach surveys, data analysis of declared intake showed significant differences between nationalities. The highest apple consumption was in Poland, where over 55% declared a consumption of more than 5 apples per week. In comparison, Italian consumers most often indicated eating 3-5 apples per week (39.3%). The lowest apple consumption was in the Netherlands and Spain. In the case of peaches, the highest consumption was indicated in France where 48% of respondents declared a peach consumption of 3-5 per week with 40% eating more than 5 fruits per week. The lowest peach intake was declared in Germany. Irrespective of country women were shown to eat more apples that men. Furthermore, the group of older people (61-70 years) consume apples more often than the adult group (36-60), while within the youngest group of consumers (16-35) eating apples was not at all popular. As with apples females declared a higher peach consumption, and again significantly lower fruit consumption by the youngest group (16-35) was indicated. Although the availability of fruit at the market remains a prime factor in determining apple and peach consumption, our survey confirmed the trends of declining this popular fruit intake by the younger generation, as well as the persistent tendency of lower frequency of fruit consumption among men than women

  8. The interdependence of the Rho GTPases and apicobasal cell polarity.

    Science.gov (United States)

    Mack, Natalie Ann; Georgiou, Marios

    2014-01-01

    Signaling via the Rho GTPases provides crucial regulation of numerous cell polarization events, including apicobasal (AB) polarity, polarized cell migration, polarized cell division and neuronal polarity. Here we review the relationships between the Rho family GTPases and epithelial AB polarization events, focusing on the 3 best-characterized members: Rho, Rac and Cdc42. We discuss a multitude of processes that are important for AB polarization, including lumen formation, apical membrane specification, cell-cell junction assembly and maintenance, as well as tissue polarity. Our discussions aim to highlight the immensely complex regulatory mechanisms that encompass Rho GTPase signaling during AB polarization. More specifically, in this review we discuss several emerging common themes, that include: 1) the need for Rho GTPase activities to be carefully balanced in both a spatial and temporal manner through a multitude of mechanisms; 2) the existence of signaling feedback loops and crosstalk to create robust cellular responses; and 3) the frequent multifunctionality that exists among AB polarity regulators. Regarding this latter theme, we provide further discussion of the potential plasticity of the cell polarity machinery and as a result the possible implications for human disease.

  9. The Rho-family GTPase Rac1 regulates integrin localization in Drosophila immunosurveillance cells.

    Directory of Open Access Journals (Sweden)

    Miguel J Xavier

    Full Text Available BACKGROUND: When the parasitoid wasp Leptopilina boulardi lays an egg in a Drosophila larva, phagocytic cells called plasmatocytes and specialized cells known as lamellocytes encapsulate the egg. The Drosophila β-integrin Myospheroid (Mys is necessary for lamellocytes to adhere to the cellular capsule surrounding L. boulardi eggs. Integrins are heterodimeric adhesion receptors consisting of α and β subunits, and similar to other plasma membrane receptors undergo ligand-dependent endocytosis. In mammalian cells it is known that integrin binding to the extracellular matrix induces the activation of Rac GTPases, and we have previously shown that Rac1 and Rac2 are necessary for a proper encapsulation response in Drosophila larvae. We wanted to test the possibility that Myospheroid and Rac GTPases interact during the Drosophila anti-parasitoid immune response. RESULTS: In the current study we demonstrate that Rac1 is required for the proper localization of Myospheroid to the cell periphery of haemocytes after parasitization. Interestingly, the mislocalization of Myospheroid in Rac1 mutants is rescued by hyperthermia, involving the heat shock protein Hsp83. From these results we conclude that Rac1 and Hsp83 are required for the proper localization of Mys after parasitization. SIGNIFICANCE: We show for the first time that the small GTPase Rac1 is required for Mysopheroid localization. Interestingly, the necessity of Rac1 in Mys localization was negated by hyperthermia. This presents a problem, in Drosophila we quite often raise larvae at 29°C when using the GAL4/UAS misexpression system. If hyperthermia rescues receptor endosomal recycling defects, raising larvae in hyperthermic conditions may mask potentially interesting phenotypes.

  10. A role for Sar1 and ARF1 GTPases during Golgi biogenesis in the protozoan parasite Trypanosoma brucei

    Science.gov (United States)

    Yavuz, Sevil; Warren, Graham

    2017-01-01

    A single Golgi stack is duplicated and partitioned into two daughter cells during the cell cycle of the protozoan parasite Trypanosoma brucei. The source of components required to generate the new Golgi and the mechanism by which it forms are poorly understood. Using photoactivatable GFP, we show that the existing Golgi supplies components directly to the newly forming Golgi in both intact and semipermeabilized cells. The movement of a putative glycosyltransferase, GntB, requires the Sar1 and ARF1 GTPases in intact cells. In addition, we show that transfer of GntB from the existing Golgi to the new Golgi can be recapitulated in semipermeabilized cells and is sensitive to the GTP analogue GTPγS. We suggest that the existing Golgi is a key source of components required to form the new Golgi and that this process is regulated by small GTPases. PMID:28495798

  11. Conservation implications of the mating system of the Pampa Hermosa landrace of peach palm analyzed with microsatellite markers.

    Science.gov (United States)

    Picanço-Rodrigues, Doriane; Astolfi-Filho, Spartaco; Lemes, Maristerra R; Gribel, Rogerio; Sebbenn, Alexandre M; Clement, Charles R

    2015-03-01

    Peach palm (Bactris gasipaes) is cultivated by many indigenous and traditional communities from Amazonia to Central America for its edible fruits, and is currently important for its heart-of-palm. The objective of this study was to investigate the mating system of peach palm, as this is important for conservation and breeding. Eight microsatellite loci were used to genotype 24 open-pollinated progenies from three populations of the Pampa Hermosa landrace maintained in a progeny trial for genetic improvement. Both the multi-locus outcrossing rates (0.95 to 0.99) and the progeny level multi-locus outcrossing rates (0.9 to 1.0) were high, indicating that peach palm is predominantly allogamous. The outcrossing rates among relatives were significantly different from zero (0.101 to 0.202), providing evidence for considerable biparental inbreeding within populations, probably due to farmers planting seeds of a small number of open-pollinated progenies in the same plot. The correlations of paternity estimates were low (0.051 to 0.112), suggesting a large number of pollen sources (9 to 20) participating in pollination of individual fruit bunches. Effective population size estimates suggest that current germplasm collections are insufficient for long-term ex situ conservation. As with most underutilized crops, on farm conservation is the most important component of an integrated conservation strategy.

  12. Conservation implications of the mating system of the Pampa Hermosa landrace of peach palm analyzed with microsatellite markers

    Directory of Open Access Journals (Sweden)

    Doriane Picanço-Rodrigues

    2015-03-01

    Full Text Available Peach palm (Bactris gasipaes is cultivated by many indigenous and traditional communities from Amazonia to Central America for its edible fruits, and is currently important for its heart-of-palm. The objective of this study was to investigate the mating system of peach palm, as this is important for conservation and breeding. Eight microsatellite loci were used to genotype 24 open-pollinated progenies from three populations of the Pampa Hermosa landrace maintained in a progeny trial for genetic improvement. Both the multi-locus outcrossing rates (0.95 to 0.99 and the progeny level multi-locus outcrossing rates (0.9 to 1.0 were high, indicating that peach palm is predominantly allogamous. The outcrossing rates among relatives were significantly different from zero (0.101 to 0.202, providing evidence for considerable biparental inbreeding within populations, probably due to farmers planting seeds of a small number of open-pollinated progenies in the same plot. The correlations of paternity estimates were low (0.051 to 0.112, suggesting a large number of pollen sources (9 to 20 participating in pollination of individual fruit bunches. Effective population size estimates suggest that current germplasm collections are insufficient for long-term ex situ conservation. As with most underutilized crops, on farm conservation is the most important component of an integrated conservation strategy.

  13. Activity of pectin methyl esterase during blanching of peaches

    NARCIS (Netherlands)

    Tijskens, L.M.M.; Rodis, P.S.; Hertog, M.L.A.T.M.; Proxenia, N.; Dijk, van C.

    1999-01-01

    The activity of pectin methyl esterase (PE) in peaches during blanching treatments was modelled and analyzed. It was postulated that the enzyme exists in two configurations, one bound and one soluble. The bound configuration can be converted into the soluble configuration. These two configurations

  14. Deficit irrigation of peach trees to reduce water consumption

    Science.gov (United States)

    Lack of water is a major limiting factor for production tree fruits such as peaches in the San Joaquin Valley of California and many other arid- or semi-arid regions in the world. Deficit irrigation can be used in some cropping systems as a water resource management strategy to reduce non-productiv...

  15. 75 FR 3694 - Radio Broadcasting Services, Peach Springs, Arizona

    Science.gov (United States)

    2010-01-22

    ...., Fletcher, Heald & Hildreth, PLC, 1300 N. 17th Street - Eleventh Floor, Arlington, Virginia 22209 (Counsel... website, http://www.bcpiweb.com >. The proposed channel at Peach Springs is part of a hybrid application and rulemaking proceeding. In the application (File No. BNPH-20091016ADO), Cochise Media Licenses, the...

  16. Apple and peach consumption habits across European countries

    NARCIS (Netherlands)

    Konopacka, D.; Jesionkowska, K.; Kruczynska, D.; Schoorl, F.W.

    2010-01-01

    The aim of this study was to gain information concerning apple and peach consumption frequency within different European countries in relation to age and gender. The survey was a part of a complex experiment with the aim of evaluating consumers’ preferences towards new varieties, and the data is

  17. Neurotrophin Promotes Neurite Outgrowth by Inhibiting Rif GTPase Activation Downstream of MAPKs and PI3K Signaling.

    Science.gov (United States)

    Tian, Xiaoxia; Yan, Huijuan; Li, Jiayi; Wu, Shuang; Wang, Junyu; Fan, Lifei

    2017-01-13

    Members of the well-known semaphorin family of proteins can induce both repulsive and attractive signaling in neural network formation and their cytoskeletal effects are mediated in part by small guanosine 5'-triphosphatase (GTPases). The aim of this study was to investigate the cellular role of Rif GTPase in the neurotrophin-induced neurite outgrowth. By using PC12 cells which are known to cease dividing and begin to show neurite outgrowth responding to nerve growth factor (NGF), we found that semaphorin 6A was as effective as nerve growth factor at stimulating neurite outgrowth in PC12 cells, and that its neurotrophic effect was transmitted through signaling by mitogen-activated protein kinases (MAPKs) and phosphatidylinositol-3-kinase (PI3K). We further found that neurotrophin-induced neurite formation in PC12 cells could be partially mediated by inhibition of Rif GTPase activity downstream of MAPKs and PI3K signaling. In conclusion, we newly identified Rif as a regulator of the cytoskeletal rearrangement mediated by semaphorins.

  18. Inoculation methods and aggressiveness of five Fusarium species against peach palm

    Directory of Open Access Journals (Sweden)

    Tiago Miguel Jarek

    2018-04-01

    Full Text Available ABSTRACT: Fusarium wilt is a major disease which affects peach palm (Bactris gasipaes Kunth.var gasipaes Henderson. This study aimed to evaluate inoculation methods and aggressiveness of isolates of five Fusarium species on peach palm. Fusarium proliferatum can infect the leaves, stem, and roots of peach palm. F. proliferatum, F. oxysporum species complex (FOSC, F. verticillioides, F. solani species complex (FSSC, and Gibberella fujikuroi species complex (GFSC are pathogenic to peach palm. The use of Fusarium-colonized ground corn for root inoculation was effective and reduced the level of damage to plants.

  19. Summary of core damage frequency from internal initiators: Peach Bottom

    International Nuclear Information System (INIS)

    Kolaczkowski, A.M.; Lambright, J.A.; Cathey, N.

    1986-01-01

    Probabilistic risk assessments (PRA) based on internal initiators are being conducted on a number of reference plants in order to provide the Nuclear Regulatory Commission (NRC) with updated information about light water reactor risk. The results of these analyses will be used by the NRC to prepare NUREG-1150 which will examine the NRC's current perception of risk. Peach Bottom has been chosen as one of the reference plants

  20. The GTPase Rab43 Controls the Anterograde ER-Golgi Trafficking and Sorting of GPCRs

    Directory of Open Access Journals (Sweden)

    Chunman Li

    2017-10-01

    Full Text Available G-protein-coupled receptors (GPCRs constitute the largest superfamily of cell-surface signaling proteins. However, mechanisms underlying their surface targeting and sorting are poorly understood. Here, we screen the Rab family of small GTPases in the surface transport of multiple GPCRs. We find that manipulation of Rab43 function significantly alters the surface presentation and signaling of all GPCRs studied without affecting non-GPCR membrane proteins. Rab43 specifically regulates the transport of nascent GPCRs from the endoplasmic reticulum (ER to the Golgi. More interestingly, Rab43 directly interacts with GPCRs in an activation-dependent fashion. The Rab43-binding domain identified in the receptors effectively converts non-GPCR membrane protein transport into a Rab43-dependent pathway. These data reveal a crucial role for Rab43 in anterograde ER-Golgi transport of nascent GPCRs, as well as the ER sorting of GPCR members by virtue of its ability to interact directly.

  1. Evaluation of peach palm (Bactris gasipaes Kunth) processed by radiation

    International Nuclear Information System (INIS)

    Silva, Priscila Vieira da

    2009-01-01

    The peach palm can be obtained from several species of palms, but the peach palm has attracted great interest by producers, as has characteristics of precocity, rusticity and tillering, producing a palm-quality differentiating it from other palmettos for their sweet flavor and yellowish . The food irradiation has been used as a treatment to ensure microbiological food safety of products to avoid infection. Its use combined with minimal processing could increase the safety and quality of minimally processed vegetables. We aimed at evaluating the effect of gamma radiation and electron beams to control bacteria; assess the physical characteristics through analysis of color and texture in peach palm in natura minimally processed and subjected to ionizing radiation stored at 8 deg C as well as evaluating the sensory characteristics. The results in the microbiological analysis showed that ionizing radiation promotes reduction of microbial load in both treatments. In the analysis of color we can conclude that among all the treatments the sample irradiated with 1.5 kGy showed more differences when compared with the other samples. Observing texture characteristics we could conclude that irradiation changed the texture of the palm, unlike the treatment by electron beams that showed no difference between samples. For the sensory analysis, the gamma radiation with dose of 1.5 kGy, induced changes in sensory properties to the attributes and overall appearance. The dose of 1 kGy caused no significant difference, so a recommended dose for the irradiation of the studied product. (author)

  2. Ras GTPases Modulate Morphogenesis, Sporulation and Cellulase Gene Expression in the Cellulolytic Fungus Trichoderma reesei

    Science.gov (United States)

    Zhang, Jiwei; Zhang, Yanmei; Zhong, Yaohua; Qu, Yinbo; Wang, Tianhong

    2012-01-01

    Background The model cellulolytic fungus Trichoderma reesei (teleomorph Hypocrea jecorina) is capable of responding to environmental cues to compete for nutrients in its natural saprophytic habitat despite its genome encodes fewer degradative enzymes. Efficient signalling pathways in perception and interpretation of environmental signals are indispensable in this process. Ras GTPases represent a kind of critical signal proteins involved in signal transduction and regulation of gene expression. In T. reesei the genome contains two Ras subfamily small GTPases TrRas1 and TrRas2 homologous to Ras1 and Ras2 from S. cerevisiae, but their functions remain unknown. Methodology/Principal Findings Here, we have investigated the roles of GTPases TrRas1 and TrRas2 during fungal morphogenesis and cellulase gene expression. We show that both TrRas1 and TrRas2 play important roles in some cellular processes such as polarized apical growth, hyphal branch formation, sporulation and cAMP level adjustment, while TrRas1 is more dominant in these processes. Strikingly, we find that TrRas2 is involved in modulation of cellulase gene expression. Deletion of TrRas2 results in considerably decreased transcription of cellulolytic genes upon growth on cellulose. Although the strain carrying a constitutively activated TrRas2G16V allele exhibits increased cellulase gene transcription, the cbh1 and cbh2 expression in this mutant still strictly depends on cellulose, indicating TrRas2 does not directly mediate the transmission of the cellulose signal. In addition, our data suggest that the effect of TrRas2 on cellulase gene is exerted through regulation of transcript abundance of cellulase transcription factors such as Xyr1, but the influence is independent of cAMP signalling pathway. Conclusions/Significance Together, these findings elucidate the functions for Ras signalling of T. reesei in cellular morphogenesis, especially in cellulase gene expression, which contribute to deciphering the

  3. The 'invisible hand': regulation of RHO GTPases by RHOGDIs.

    Science.gov (United States)

    Garcia-Mata, Rafael; Boulter, Etienne; Burridge, Keith

    2011-07-22

    The 'invisible hand' is a term originally coined by Adam Smith in The Theory of Moral Sentiments to describe the forces of self-interest, competition and supply and demand that regulate the resources in society. This metaphor continues to be used by economists to describe the self-regulating nature of a market economy. The same metaphor can be used to describe the RHO-specific guanine nucleotide dissociation inhibitor (RHOGDI) family, which operates in the background, as an invisible hand, using similar forces to regulate the RHO GTPase cycle.

  4. The invisible hand: regulation of RHO GTPases by RHOGDIs

    Science.gov (United States)

    Garcia-Mata, Rafael; Boulter, Etienne; Burridge, Keith

    2011-01-01

    Preface The 'invisible hand' is a term originally coined by Adam Smith in the Theory of Moral Sentiments to describe the forces of self-interest, competition, and supply and demand that regulate the resources in society. This metaphor continues to be used by economists to describe the self-regulating nature of a market economy. The same metaphor can be used to describe the RHO-specific guanine nucleotide dissociation inhibitor (RHOGDI) family, which operates in the background, as an invisible hand, using similar forces to regulate the RHO GTPase cycle. PMID:21779026

  5. Molecular characterization of peach [Prunus persica (L.) Batsch] germplasm in the United States using microsatellite markers

    Science.gov (United States)

    Peach [Prunus persica (L.) Batsch] is an important medicinal fruit with immense health benefits and antioxidant activity. In this study, microsatellite markers were used as DNA fingerprinting tools for the identification and characterization of peach germplasm in the United States. Eleven microsatel...

  6. Effects of infrared radiation heating on peeling performance and quality attributes of clingstone peaches

    Science.gov (United States)

    Salinity and wastewater disposal problems associated with the conventional wet-lye method for peeling clingstone peaches result in considerable negative environment impacts. This study investigated the efficacy of using infrared (IR) heating as an alternative method for peach peel removal without us...

  7. Identification of the GTPase superfamily in Mycoplasma synoviae and Mycoplasma hyopneumoniae

    Directory of Open Access Journals (Sweden)

    Clayton Luiz Borges

    2007-01-01

    Full Text Available Mycoplasmas are the smallest known prokaryotes with self-replication ability. They are obligate parasites, taking up many molecules of their hosts and acting as pathogens in men, animals, birds and plants. Mycoplasma hyopneumoniae is the infective agent of swine mycoplasmosis and Mycoplasma synoviae is responsible for subclinical upper respiratory infections that may result in airsacculitis and synovitis in chickens and turkeys. These highly infectious organisms present a worldwide distribution and are responsible for major economic problems. Proteins of the GTPase superfamily occur in all domains of life, regulating functions such as protein synthesis, cell cycle and differentiation. Despite their functional diversity, all GTPases are believed to have evolved from a single common ancestor. In this work we have identified mycoplasma GTPases by searching the complete genome databases of Mycoplasma synoviae and Mycoplasma hyopneumoniae, J (non-pathogenic and 7448 (pathogenic strains. Fifteen ORFs encoding predicted GTPases were found in M. synoviae and in the two strains of M. hyopneumoniae. Searches for conserved G domains in GTPases were performed and the sequences were classified into families. The GTPase phylogenetic analysis showed that the subfamilies were well resolved into clades. The presence of GTPases in the three strains suggests the importance of GTPases in 'minimalist' genomes.

  8. Human Mammary Epithelial Cell Transformation by Rho GTPase Through a Novel Mechanism

    Science.gov (United States)

    2009-08-01

    87: 635-44. 18. Burbelo P, Wellstein A, Pestell RG. Altered Rho GTPase signaling pathways in breast cancer cells. Breast Cancer Res Treat 2004; 84...Burbelo P, Wellstein A, Pestell RG. Altered Rho GTPase signaling pathways in breast cancer cells. Breast Cancer Res Treat 2004;84:43–8. 19. Band V

  9. Investigation of the aroma of commercial peach (Prunus persica L. Batsch) types by Proton Transfer Reaction-Mass Spectrometry (PTR-MS) and sensory analysis

    NARCIS (Netherlands)

    Cardoso Ferreira Pinhancos de Bianchi, Tiago; Weesepoel, Yannick; Koot, Alex; Iglesias, Ignasi; Eduardo, Iban; Gratacós-Cubarsí, Marta; Guerrero, Luis; Hortós, Maria; Ruth, van Saskia

    2017-01-01

    The aim of this study was to investigate the aroma and sensory profiles of various types of peaches (Prunus persica L. Batsch.). Forty-three commercial cultivars comprising peaches, flat peaches, nectarines, and canning peaches (pavías) were grown over two consecutive harvest years. Fruits were

  10. Rab4GTPase modulates CFTR function by impairing channel expression at plasma membrane

    International Nuclear Information System (INIS)

    Saxena, Sunil K.; Kaur, Simarna; George, Constantine

    2006-01-01

    Cystic fibrosis (CF), an autosomal recessive disorder, is caused by the disruption of biosynthesis or the function of a membrane cAMP-activated chloride channel, CFTR. CFTR regulatory mechanisms include recruitment of channel proteins to the cell surface from intracellular pools and by protein-protein interactions. Rab proteins are small GTPases involved in regulated trafficking controlling vesicle docking and fusion. Rab4 controls recycling events from endosome to the plasma membrane, fusion, and degradation. The colorectal cell line HT-29 natively expresses CFTR and responds to cAMP stimulation with an increase in CFTR-mediated currents. Rab4 over-expression in HT-29 cells inhibits both basal and cAMP-stimulated CFTR-mediated currents. GTPase-deficient Rab4Q67L and GDP locked Rab4S22N both inhibit channel activity, which appears characteristically different. Active status of Rab4 was confirmed by GTP overlay assay, while its expression was verified by Western blotting. The pull-down and immunoprecipitation experiments suggest that Rab4 physically interacts with CFTR through protein-protein interaction. Biotinylation with cell impermeant NHS-Sulfo-SS-Biotin implies that Rab4 impairs CFTR expression at cell surface. The enhanced cytosolic CFTR indicates that Rab4 expression restrains CFTR appearance at the cell membrane. The study suggests that Rab4 regulates the channel through multiple mechanisms that include protein-protein interaction, GTP/GDP exchange, and channel protein trafficking. We propose that Rab4 is a dynamic molecule with a significant role in CFTR function

  11. Retrotransposition and mutation events yield Rap1 GTPases with differential signalling capacity

    Directory of Open Access Journals (Sweden)

    Penzkofer Tobias

    2010-02-01

    Full Text Available Abstract Background Retrotransposition of mRNA transcripts gives occasionally rise to functional retrogenes. Through acquiring tempero-spatial expression patterns distinct from their parental genes and/or functional mutations in their coding sequences, such retrogenes may in principle reshape signalling networks. Results Here we present evidence for such a scenario, involving retrogenes of Rap1 belonging to the Ras family of small GTPases. We identified two murine and one human-specific retrogene of Rap1A and Rap1B, which encode proteins that differ by only a few amino acids from their parental Rap1 proteins. Markedly, human hRap1B-retro and mouse mRap1A-retro1 acquired mutations in the 12th and 59th amino acids, respectively, corresponding to residues mutated in constitutively active oncogenic Ras proteins. Statistical and structural analyses support a functional evolution scenario, where Rap1 isoforms of retrogenic origin are functionally distinct from their parental proteins. Indeed, all retrogene-encoded GTPases have an increased GTP/GDP binding ratio in vivo, indicating that their conformations resemble that of active GTP-bound Rap1. We furthermore demonstrate that these three Rap1 isoforms exhibit distinct affinities for the Ras-binding domain of RalGDS. Finally, when tested for their capacity to induce key cellular processes like integrin-mediated cell adhesion or cell spreading, marked differences are seen. Conclusions Together, these data lend strong support for an evolution scenario, where retrotransposition and subsequent mutation events generated species-specific Rap1 isoforms with differential signaling potential. Expression of the constitutively active human Rap1B-retro in cells like those derived from Ramos Burkitt's lymphoma and bone marrow from a patient with myelodysplastic syndrome (MDS warrants further investigation into its role in disease development.

  12. RhoA GTPase regulates radiation-induced alterations in endothelial cell adhesion and migration

    International Nuclear Information System (INIS)

    Rousseau, Matthieu; Gaugler, Marie-Hélène; Rodallec, Audrey; Bonnaud, Stéphanie; Paris, François; Corre, Isabelle

    2011-01-01

    Highlights: ► We explore the role of RhoA in endothelial cell response to ionizing radiation. ► RhoA is rapidly activated by single high-dose of radiation. ► Radiation leads to RhoA/ROCK-dependent actin cytoskeleton remodeling. ► Radiation-induced apoptosis does not require the RhoA/ROCK pathway. ► Radiation-induced alteration of endothelial adhesion and migration requires RhoA/ROCK. -- Abstract: Endothelial cells of the microvasculature are major target of ionizing radiation, responsible of the radiation-induced vascular early dysfunctions. Molecular signaling pathways involved in endothelial responses to ionizing radiation, despite being increasingly investigated, still need precise characterization. Small GTPase RhoA and its effector ROCK are crucial signaling molecules involved in many endothelial cellular functions. Recent studies identified implication of RhoA/ROCK in radiation-induced increase in endothelial permeability but other endothelial functions altered by radiation might also require RhoA proteins. Human microvascular endothelial cells HMEC-1, either treated with Y-27632 (inhibitor of ROCK) or invalidated for RhoA by RNA interference were exposed to 15 Gy. We showed a rapid radiation-induced activation of RhoA, leading to a deep reorganisation of actin cytoskeleton with rapid formation of stress fibers. Endothelial early apoptosis induced by ionizing radiation was not affected by Y-27632 pre-treatment or RhoA depletion. Endothelial adhesion to fibronectin and formation of focal adhesions increased in response to radiation in a RhoA/ROCK-dependent manner. Consistent with its pro-adhesive role, ionizing radiation also decreased endothelial cells migration and RhoA was required for this inhibition. These results highlight the role of RhoA GTPase in ionizing radiation-induced deregulation of essential endothelial functions linked to actin cytoskeleton.

  13. Comprehensive functional analysis of Rab GTPases in Drosophila nephrocytes.

    Science.gov (United States)

    Fu, Yulong; Zhu, Jun-Yi; Zhang, Fujian; Richman, Adam; Zhao, Zhanzheng; Han, Zhe

    2017-06-01

    The Drosophila nephrocyte is a critical component of the fly renal system and bears structural and functional homology to podocytes and proximal tubule cells of the mammalian kidney. Investigations of nephrocyte cell biological processes are fundamental to understanding the insect renal system. Nephrocytes are highly active in endocytosis and vesicle trafficking. Rab GTPases regulate endocytosis and trafficking but specific functions of nephrocyte Rabs remain undefined. We analyzed Rab GTPase expression and function in Drosophila nephrocytes and found that 11 out of 27 Drosophila Rabs were required for normal activity. Rabs 1, 5, 7, 11 and 35 were most important. Gene silencing of the nephrocyte-specific Rab5 eliminated all intracellular vesicles and the specialized plasma membrane structures essential for nephrocyte function. Rab7 silencing dramatically increased clear vacuoles and reduced lysosomes. Rab11 silencing increased lysosomes and reduced clear vacuoles. Our results suggest that Rab5 mediates endocytosis that is essential for the maintenance of functionally critical nephrocyte plasma membrane structures and that Rabs 7 and 11 mediate alternative downstream vesicle trafficking pathways leading to protein degradation and membrane recycling, respectively. Elucidating molecular pathways underlying nephrocyte function has the potential to yield important insights into human kidney cell physiology and mechanisms of cell injury that lead to disease. The Drosophila nephrocyte is emerging as a useful in vivo model system for molecular target identification and initial testing of therapeutic approaches in humans.

  14. Characterization of mitochondrion-targeted GTPases in Plasmodium falciparum.

    Science.gov (United States)

    Gupta, Kirti; Gupta, Ankit; Haider, Afreen; Habib, Saman

    2018-04-12

    Ribosome assembly is critical for translation and regulating the response to cellular events and requires a complex interplay of ribosomal RNA and proteins with assembly factors. We investigated putative participants in the biogenesis of the reduced organellar ribosomes of Plasmodium falciparum and identified homologues of two assembly GTPases - EngA and Obg that were found in mitochondria. Both are indispensable in bacteria and P. berghei EngA is among the 'essential' parasite blood stage proteins identified recently. PfEngA and PfObg1 interacted with parasite mitoribosomes in vivo. GTP stimulated PfEngA interaction with the 50S subunit of Escherichia coli surrogate ribosomes. Although PfObg1-ribosome interaction was independent of nucleotide binding, GTP hydrolysis by PfObg1 was enhanced upon ribosomal association. An additional function for PfObg1 in mitochondrial DNA transactions was suggested by its specific interaction with the parasite mitochondrial genome in vivo. Deletion analysis revealed that the positively-charged OBG (spoOB-associated GTP-binding protein) domain mediates DNA-binding. A role for PfEngA in mitochondrial genotoxic stress response was indicated by its over-expression upon methyl methanesulfonate-induced DNA damage. PfEngA had lower sensitivity to an E. coli EngA inhibitor suggesting differences with bacterial counterparts. Our results show the involvement of two important GTPases in P. falciparum mitochondrial function, with the first confirmed localization of an EngA homologue in eukaryotic mitochondria.

  15. Ultrasound Assisted Extraction of Phenolic Compounds from Peaches and Pumpkins

    Science.gov (United States)

    Altemimi, Ammar; Watson, Dennis G.; Choudhary, Ruplal; Dasari, Mallika R.; Lightfoot, David A.

    2016-01-01

    The ultrasound-assisted extraction (UAE) method was used to optimize the extraction of phenolic compounds from pumpkins and peaches. The response surface methodology (RSM) was used to study the effects of three independent variables each with three treatments. They included extraction temperatures (30, 40 and 50°C), ultrasonic power levels (30, 50 and 70%) and extraction times (10, 20 and 30 min). The optimal conditions for extractions of total phenolics from pumpkins were inferred to be a temperature of 41.45°C, a power of 44.60% and a time of 25.67 min. However, an extraction temperature of 40.99°C, power of 56.01% and time of 25.71 min was optimal for recovery of free radical scavenging activity (measured by 1, 1-diphenyl-2-picrylhydrazyl (DPPH) reduction). The optimal conditions for peach extracts were an extraction temperature of 41.53°C, power of 43.99% and time of 27.86 min for total phenolics. However, an extraction temperature of 41.60°C, power of 44.88% and time of 27.49 min was optimal for free radical scavenging activity (judged by from DPPH reduction). Further, the UAE processes were significantly better than solvent extractions without ultrasound. By electron microscopy it was concluded that ultrasonic processing caused damage in cells for all treated samples (pumpkin, peach). However, the FTIR spectra did not show any significant changes in chemical structures caused by either ultrasonic processing or solvent extraction. PMID:26885655

  16. Peach Bottom transient analysis with BWR TRACB02

    International Nuclear Information System (INIS)

    Alamgir, M.; Sutherland, W.A.

    1984-01-01

    TRAC calculations have been performed for a Turbine Trip transient (TT1) in the Peach Bottom BWR power plant. This study is a part of the qualification of the BWR-TRAC code. The simulation is aimed at reproducing the observed thermal hydraulic behavior in a pressurization transient. Measured core power is an input to the calculation. Comparison with data show the code reasonably well predicts the generation and propagation of the pressure waves in the main steam line and associated pressurization of the reactor vessel following the closure of the turbine stop valve

  17. Formulation of a peach ice cream as potential symbiotic food

    OpenAIRE

    VILLALVA, Fernando Josué; CRAVERO BRUNERI, Andrea Paula; VINDEROLA, Gabriel; GONÇALVEZ DE OLIVEIRA, Enzo; PAZ, Noelia Fernanda; RAMÓN, Adriana Noemí

    2017-01-01

    Abstract Today’s population increasingly demands and consumes healthy products. For this reason, the food industry has been developing and marketing food with added bioactive components. The aim of this work was to formulate a peach ice cream reduced in calories with an added probiotic (Bifidobacterium lactis Bb-12) and prebiotics (inulin), and to evaluate its sensory quality and acceptability as potential symbiotic food. The moisture content was 76.47%; 7.14% protein; 0.15% fat; 6.37%; carbo...

  18. PlexinA2 Forward Signaling through Rap1 GTPases Regulates Dentate Gyrus Development and Schizophrenia-like Behaviors

    Directory of Open Access Journals (Sweden)

    Xiao-Feng Zhao

    2018-01-01

    Full Text Available Summary: Dentate gyrus (DG development requires specification of granule cell (GC progenitors in the hippocampal neuroepithelium, as well as their proliferation and migration into the primordial DG. We identify the Plexin family members Plxna2 and Plxna4 as important regulators of DG development. Distribution of immature GCs is regulated by Sema5A signaling through PlxnA2 and requires a functional PlxnA2 GTPase-activating protein (GAP domain and Rap1 small GTPases. In adult Plxna2−/− but not Plxna2-GAP-deficient mice, the dentate GC layer is severely malformed, neurogenesis is compromised, and mossy fibers form aberrant synaptic boutons within CA3. Behavioral studies with Plxna2−/− mice revealed deficits in associative learning, sociability, and sensorimotor gating—traits commonly observed in neuropsychiatric disorder. Remarkably, while morphological defects are minimal in Plxna2-GAP-deficient brains, defects in fear memory and sensorimotor gating persist. Since allelic variants of human PLXNA2 and RAP1 associate with schizophrenia, our studies identify a biochemical pathway important for brain development and mental health. : Zhao et al. find that Sema5A-PlexinA2 forward signaling through Rap1 GTPases is required for progenitor distribution in the developing mouse dentate gyrus. Adult Plxna2−/−, but not Plxna2-GAP-deficient, mice show defects in dentate morphology, neurogenesis, and mossy fiber connectivity. Plxna2−/− and Plxna2-GAP mice exhibit behavioral defects suggestive of neuropsychiatric illness. Keywords: PlexinA2, semaphoring, Rap1, GAP, dentate gyrus, adult neurogenesis, mossy fiber, fear memory, sensorimotor gating, schizophrenia

  19. A bottom-up model to describe consumers’ preferences towards late season peaches

    Energy Technology Data Exchange (ETDEWEB)

    Groot, E.; Albisu, L.M.

    2015-07-01

    Peaches are consumed in Mediterranean countries since ancient times. Nowadays there are few areas in Europe that produce peaches with Protected Designation of Origin (PDO), and the Calanda area is one of them. The aim of this work is to describe consumers’ preferences towards late season PDO Calanda peaches in the city of Zaragoza, Spain, by a bottom-up model. The bottom-up model proves greater amount of information than top-down models. In this approach it is estimated one utility function per consumer. Thus, it is not necessary to make assumptions about preference distributions and correlations across respondents. It was observed that preference distributions were neither normal nor independently distributed. If those preferences were estimated by top-down models, conclusions would be biased. This paper also explores a new way to describe preferences through individual utility functions. Results show that the largest behavioural group gathered origin sensitive consumers. Their utility increased if the peaches were produced in the Calanda area and, especially, when peaches had the PDO Calanda brand. In sequence, the second most valuable attribute for consumers was the price. Peach size and packaging were not so important on purchase choice decision. Nevertheless, it is advisable to avoid trading smallest size peaches (weighting around 160 g/fruit). Traders also have to be careful by using active packaging. It was found that a group of consumers disliked this kind of product, probably, because they perceived it as less natural. (Author)

  20. Impact of glutathione on the allergenicity of the peach lipid transfer protein Pru p 3.

    Science.gov (United States)

    Gómez-Casado, C; Tordesillas, L; Kinkel, J; Starkl, P; Cuesta-Herranz, J; Roth-Walter F; Díaz-Perales, A; Jensen-Jarolim, E

    2015-01-01

    The allergenic potential of proteins can be altered under various physicochemical conditions. Glutathione (GSH) is a reducing agent that is used as an antioxidant in food products. We aimed to characterize the natural folding of peach proteins and test the allergenicity of reduced and natural Pru p 3, the major peach allergen. Pru p 3 was purified from peach, and its conformation was analyzed by means of circular dichroism. Using a thiol fluorescent probe, reduced proteins were detected in fresh peach. GSH-reduced Pru p 3 was tested in vitro for T-cell proliferation and in vivo using skin prick testing. GSH-reduced Pru p 3 produced variable skin prick reactions in peach-allergic patients. The proliferative response of peripheral blood mononuclear cells from allergic patients to reduced Pru p 3 tended to be less intense, whereas secretion of the cytokines IFN-γ, IL-5, and IL-10 was comparable. In a pool of sera from peach-allergic patients, reduction hardly impaired IgE-binding. Moreover, the stability of reduced Pru p 3 to gastrointestinal digestion was similar to that of the natural form. GSH can at least transiently reduce Pru p 3. We found that the effect of reduction on the allergenicity of Pru p 3 varied. Therefore, as an additive, GSH does not seem to eliminate the risk of reactions for peach-allergic patients.

  1. Formulation of a peach ice cream as potential symbiotic food

    Directory of Open Access Journals (Sweden)

    Fernando Josué VILLALVA

    Full Text Available Abstract Today’s population increasingly demands and consumes healthy products. For this reason, the food industry has been developing and marketing food with added bioactive components. The aim of this work was to formulate a peach ice cream reduced in calories with an added probiotic (Bifidobacterium lactis Bb-12 and prebiotics (inulin, and to evaluate its sensory quality and acceptability as potential symbiotic food. The moisture content was 76.47%; 7.14% protein; 0.15% fat; 6.37%; carbohydrates; 9.87% inulin; 1.22% ash; 0.201% calcium, 0.155% phosphorus and 0.168% sodium. On the first and 21th day of storage counts of B. lactis Bb – 12 was 4 x 108 CFU/mL and 1.5 x 107 CFU/mL, respectively. It was possible to formulate a peach ice cream reduced in calories, fat, and sugar and with potential symbiotic effect, by addition of B. lactis Bb – 12. A product with suitable organoleptic characteristics, creamy texture, peachy colour, taste and flavour, and no ice crystals was obtained. This ice cream would be a suitable food matrix to incorporate prebiotic and probiotic ingredients as a potential symbiotic food.

  2. Low dose UV and gamma radiation on storage rot and physicochemical changes in peaches

    International Nuclear Information System (INIS)

    Lu, J.Y.; Lukombo, S.M.; Stevens, C.; Khan, V.A.; Wilson, C.L.; Pusey, P.L.; Chaultz, E.

    1993-01-01

    Peach fruit were irradiated with 7.5 x 10(4) ergs/mm(2) of UV (254nm) or 0.1 kGy gamma rays or a combination of both, then stored at 16C for 21 days. The results showed that both UV and gamma rays reduced storage rot and delayed ripening. UV treated peaches had lower sugar concentration, total phenols, anthocyanins and lower weight loss than the gamma treated peaches. The combination of UV and gamma showed no advantage over the use of UV or gamma alone

  3. Rab GTPases and the Autophagy Pathway: Bacterial Targets for a Suitable Biogenesis and Trafficking of Their Own Vacuoles

    Directory of Open Access Journals (Sweden)

    María Milagros López de Armentia

    2016-03-01

    Full Text Available Autophagy is an intracellular process that comprises degradation of damaged organelles, protein aggregates and intracellular pathogens, having an important role in controlling the fate of invading microorganisms. Intracellular pathogens are internalized by professional and non-professional phagocytes, localizing in compartments called phagosomes. To degrade the internalized microorganism, the microbial phagosome matures by fusion events with early and late endosomal compartments and lysosomes, a process that is regulated by Rab GTPases. Interestingly, in order to survive and replicate in the phagosome, some pathogens employ different strategies to manipulate vesicular traffic, inhibiting phagolysosomal biogenesis (e.g., Staphylococcus aureus and Mycobacterium tuberculosis or surviving in acidic compartments and forming replicative vacuoles (e.g., Coxiella burnetti and Legionella pneumophila. The bacteria described in this review often use secretion systems to control the host’s response and thus disseminate. To date, eight types of secretion systems (Type I to Type VIII are known. Some of these systems are used by bacteria to translocate pathogenic proteins into the host cell and regulate replicative vacuole formation, apoptosis, cytokine responses, and autophagy. Herein, we have focused on how bacteria manipulate small Rab GTPases to control many of these processes. The growing knowledge in this field may facilitate the development of new treatments or contribute to the prevention of these types of bacterial infections.

  4. 76 FR 31295 - Nectarines and Peaches Grown in California; Notice of Withdrawal

    Science.gov (United States)

    2011-05-31

    ... forms to collect information related to the Federal marketing orders for nectarines and peaches grown in... FURTHER INFORMATION CONTACT: Andrew Hatch, Supervisory Marketing Specialist, Marketing Order... Email: [email protected] . SUPPLEMENTARY INFORMATION: Marketing Order Nos. 916 and 917...

  5. Hyperspectral imaging detection of decayed honey peaches based on their chlorophyll content.

    Science.gov (United States)

    Sun, Ye; Wang, Yihang; Xiao, Hui; Gu, Xinzhe; Pan, Leiqing; Tu, Kang

    2017-11-15

    Honey peach is a very common but highly perishable market fruit. When pathogens infect fruit, chlorophyll as one of the important components related to fruit quality, decreased significantly. Here, the feasibility of hyperspectral imaging to determine the chlorophyll content thus distinguishing diseased peaches was investigated. Three optimal wavelengths (617nm, 675nm, and 818nm) were selected according to chlorophyll content via successive projections algorithm. Partial least square regression models were established to determine chlorophyll content. Three band ratios were obtained using these optimal wavelengths, which improved spatial details, but also integrates the information of chemical composition from spectral characteristics. The band ratio values were suitable to classify the diseased peaches with 98.75% accuracy and clearly show the spatial distribution of diseased parts. This study provides a new perspective for the selection of optimal wavelengths of hyperspectral imaging via chlorophyll content, thus enabling the detection of fungal diseases in peaches. Copyright © 2017 Elsevier Ltd. All rights reserved.

  6. 7 CFR 319.56-23 - Apricots, nectarines, peaches, plumcot, and plums from Chile.

    Science.gov (United States)

    2010-01-01

    ... organization of Chile (Servicio Agricola y Ganadero, referred to in this section as SAG) or a private export...) Responsibilities of Servicio Agricola y Ganadero. SAG will ensure that: (1) Apricots, nectarines, peaches, plumcot...

  7. Coupling mechanical tension and GTPase signaling to generate cell and tissue dynamics

    Science.gov (United States)

    Zmurchok, Cole; Bhaskar, Dhananjay; Edelstein-Keshet, Leah

    2018-07-01

    Regulators of the actin cytoskeleton such Rho GTPases can modulate forces developed in cells by promoting actomyosin contraction. At the same time, through mechanosensing, tension is known to affect the activity of Rho GTPases. What happens when these effects act in concert? Using a minimal model (1 GTPase coupled to a Kelvin–Voigt element), we show that two-way feedback between signaling (‘RhoA’) and mechanical tension (stretching) leads to a spectrum of cell behaviors, including contracted or relaxed cells, and cells that oscillate between these extremes. When such ‘model cells’ are connected to one another in a row or in a 2D sheet (‘epithelium’), we observe waves of contraction/relaxation and GTPase activity sweeping through the tissue. The minimal model lends itself to full bifurcation analysis, and suggests a mechanism that explains behavior observed in the context of development and collective cell behavior.

  8. Sensory evaluation of Regina freestone peaches treated with low doses of gamma radiation

    International Nuclear Information System (INIS)

    O'Mahony, M.; Wong, S.Y.; Odbert, N.

    1985-01-01

    Sensory appraisal of low post-harvest gamma irradiation dosing (65-75 Krad) of a single batch of peaches revealed significant differences in aroma and in taste components not associated with sweetness, but only slight differences in firmness and appearance. A panel of practiced judges evaluated irradiated and non-irradiated peaches using a technique of minimal cross-sensory inference. The significant differences in aroma and taste also were detected by untrained judges

  9. Genetic dissection of Sharka disease tolerance in peach (P. persica L. Batsch).

    Science.gov (United States)

    Cirilli, Marco; Rossini, Laura; Geuna, Filippo; Palmisano, Francesco; Minafra, Angelantonio; Castrignanò, Tiziana; Gattolin, Stefano; Ciacciulli, Angelo; Babini, Anna Rosa; Liverani, Alessandro; Bassi, Daniele

    2017-11-03

    Plum pox virus (PPV), agent of Sharka disease, is the most important quarantine pathogen of peach (P. persica L. Batsch). Extensive evaluation of peach germplasm has highlighted the lack of resistant sources, while suggesting the presence of a quantitative disease resistance, expressed as reduction in the intensity of symptoms. Unravelling the genetic architecture of peach response to PPV infection is essential for pyramiding resistant genes and for developing more tolerant varieties. For this purpose, a genome-wide association (GWA) approach was applied in a panel of accessions phenotyped for virus susceptibility and genotyped with the IPSC peach 9 K SNP Array, and coupled with an high-coverage resequencing of the tolerant accession 'Kamarat'. Genome-wide association identified three highly significant associated loci on chromosome 2 and 3, accounting for most of the reduction in PPV-M susceptibility within the analysed peach population. The exploration of associated intervals through whole-genome comparison of the tolerant accession 'Kamarat' and other susceptible accessions, including the PPV-resistant wild-related species P. davidiana, allow the identification of allelic variants in promising candidate genes, including an RTM2-like gene already characterized in A. thaliana. The present study is the first effort to identify genetic factors involved in Sharka disease in peach germplasm through a GWA approach. We provide evidence of the presence of quantitative resistant loci in a collection of peach accessions, identifying major loci and highly informative SNPs that could be useful for marker assisted selection. These results could serve as reference bases for future research aimed at the comprehension of genetic mechanism regulating the complex peach-PPV interaction.

  10. Rho GTPases, their post-translational modifications, disease-associated mutations and pharmacological inhibitors.

    Science.gov (United States)

    Olson, Michael F

    2018-05-04

    The 20 members of the Rho GTPase family are key regulators of a wide-variety of biological activities. In response to activation, they signal via downstream effector proteins to induce dynamic alterations in the organization of the actomyosin cytoskeleton. In this review, post-translational modifications, mechanisms of dysregulation identified in human pathological conditions, and the ways that Rho GTPases might be targeted for chemotherapy will be discussed.

  11. Vesicular Trafficking Defects, Developmental Abnormalities, and Alterations in the Cellular Death Process Occur in Cell Lines that Over-Express Dictyostelium GTPase, Rab2, and Rab2 Mutants

    Directory of Open Access Journals (Sweden)

    Katherine Maringer

    2014-08-01

    Full Text Available Small molecular weight GTPase Rab2 has been shown to be a resident of pre-Golgi intermediates and required for protein transport from the ER to the Golgi complex, however, the function of Rab2 in Dictyostelium has yet to be fully characterized. Using cell lines that over-express DdRab2, as well as cell lines over-expressing constitutively active (CA, and dominant negative (DN forms of the GTPase, we report a functional role in vesicular transport specifically phagocytosis, and endocytosis. Furthermore, Rab2 like other GTPases cycles between an active GTP-bound and an inactive GDP-bound state. We found that this GTP/GDP cycle for DdRab2 is crucial for normal Dictyostelium development and cell–cell adhesion. Similar to Rab5 and Rab7 in C. elegans, we found that DdRab2 plays a role in programmed cell death, possibly in the phagocytic removal of apoptotic corpses.

  12. Risk-based selection of SSCs at Peach Bottom

    International Nuclear Information System (INIS)

    Krueger, G.A.; Marie, A.J.

    1993-01-01

    The purpose of identifying risk significant systems, structures, and components (SSCS) that are within the scope of the maintenance rule is to bring a higher level of attention to a subset of those SSCS. These risk-significant SSCs will have specific performance criteria established for them, and failure to meet this performance criteria will result in establishing goals to ensure the necessary improvement in performance. The Peach Bottom individual plant examination (IPE) results were used to provide insights for the verification of proposed probabilistic risk assessment (PRA) methods set forth in the Industry Maintenance Guidelines for Implementation of the Maintenance Rule. The objective of reviewing the methods for selection of SSCs that are considered risk significant was to ensure the methods used are logical, reproducible, and can be consistently applied

  13. Analysis of Peach Bottom station blackout with MELCOR

    International Nuclear Information System (INIS)

    Dingman, S.E.; Cole, R.K.; Haskin, F.E.; Summers, R.M.; Webb, S.W.

    1987-01-01

    A demonstration analysis of station blackout at Peach Bottom has been performed using MELCOR and the results have been compared with those from MARCON 2.1B and the Source Term Code Package (STCP). MELCOR predicts greater in-vessel hydrogen production, earlier melting and core collapse, but later debris discharge than MARCON 2.1B. The drywell fails at vessel breach in MELCOR, but failure is delayed about an hour in MARCON 2.1B. These differences are mainly due to the MELCOR models for candling during melting, in-core axial conduction, and continued oxidation and heat transfer from core debris following lower head dryout. Three sensitivity calculations have been performed with MELCOR to address uncertainties regarding modeling of the core-concrete interactions. The timing of events and the gas and radionuclide release rates are somewhat different in the base case and the three sensitivity cases, but the final conditions and total releases are similar

  14. Implications of Rho GTPase signaling in glioma cell invasion and tumor progression

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    Shannon Patricia Fortin Ensign

    2013-10-01

    Full Text Available Glioblastoma (GB is the most malignant of primary adult brain tumors, characterized by a highly locally-invasive cell population, as well as abundant proliferative cells, neoangiogenesis, and necrosis. Clinical intervention with chemotherapy or radiation may either promote or establish an environment for manifestation of invasive behavior. Understanding the molecular drivers of invasion in the context of glioma progression may be insightful in directing new treatments for patients with GB. Here, we review current knowledge on Rho family GTPases, their aberrant regulation in GB, and their effect on GB cell invasion and tumor progression. Rho GTPases are modulators of cell migration through effects on actin cytoskeleton rearrangement; in non-neoplastic tissue, expression and activation of Rho GTPases are normally under tight regulation. In GB, Rho GTPases are deregulated, often via hyperactivity or overexpression of their activators, Rho GEFs. Downstream effectors of Rho GTPases have been shown to promote invasiveness and, importantly, glioma cell survival. The study of aberrant Rho GTPase signaling in GB is thus an important investigation of cell invasion as well as treatment resistance and disease progression.

  15. Effect of Particle Orientation during Thermal Processing of Canned Peach Halves: A CFD Simulation

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    Adreas Dimou

    2014-05-01

    Full Text Available The objective of this work was to apply Computational Fluid Dynamics (CFD to study the effect of particle orientation on fluid flow, temperature evolution, as well as microbial destruction, during thermal processing of still cans filled with peach halves in sugar syrup. A still metal can with four peach halves in 20% sugar syrup was heated at 100 °C for 20 min and thereafter cooled at 20 °C. Infinite heat transfer coefficient between heating medium and external can wall was considered. Peach halves were orderly placed inside the can with the empty space originally occupied by the kernel facing, in all peaches, either towards the top or the bottom of the can. In a third situation, the can was placed horizontally. Simulations revealed differences on particle temperature profiles, as well as process F values and critical point location, based on their orientation. At their critical points, peach halves with the kernel space facing towards the top of the can heated considerably slower and cooled faster than the peaches having their kernel space facing towards the bottom of the can. The horizontal can case exhibited intermediate cooling but the fastest heating rates and the highest F process values among the three cases examined. The results of this study could be used in designing of thermal processes with optimal product quality.

  16. Silage or fresh by-product of peach palm as roughage in the feeding of lambs.

    Science.gov (United States)

    dos Santos Cabral, Ícaro; Azevêdo, José Augusto Gomes; de Almeida, Flávio Moreira; Pereira, Luiz Gustavo Ribeiro; de Araújo, Gherman Garcia Leal; Nogueira, Abdon Santos; Souza, Lígia Lins; de Oliveira, Gisele Andrade; de Oliveira Filho, Carlos Alberto Alves

    2015-03-01

    The objective of this study was to evaluate intake and apparent digestibility of agro-industrial by-product of peach palm in diets for lambs. Twenty castrated, crossbred Santa Ines lambs, with average age of 150 days and body weight of 22.4 ± 3.4 kg, were distributed in a completely randomized design with four experimental diets composed of the following: fresh by-product of peach palm enriched with urea + ammonia sulfate (FU); fresh peach palm by-product + concentrate (FP); silage of peach palm by-product + concentrate (SP); and silage of peach palm by-product enriched with 15% of cornmeal + concentrate (SPC). Intake was recorded daily, and the digestibility coefficients were estimated with the internal marker indigestible acid detergent fiber (iADF). Diet FU resulted in the lowest intake and digestibility of the nutrients evaluated. Animals receiving diet FP showed higher intakes of dry matter (DM), organic matter (OM), crude protein (CP), neutral detergent fiber (NDF), total digestible nutrients (TDN), and digestible energy (DE) in relation to animals fed diets SP and SPC. Diets SP and SPC showed higher coefficients of digestibility of DM, OM, CP, and NDF than diet FP. Diet SP reduced the intakes of DM, OM, ether extract (EE), non-fibrous carbohydrate (NFC), TDN, and DE and the digestibility coefficients of DM, OM, and NFC as compared with diet SPC. Feedlot lambs fed a diet with fresh peach palm by-product + concentrate (diet FP) have higher nutrient intake.

  17. Candidate gene database and transcript map for peach, a model species for fruit trees.

    Science.gov (United States)

    Horn, Renate; Lecouls, Anne-Claire; Callahan, Ann; Dandekar, Abhaya; Garay, Lilibeth; McCord, Per; Howad, Werner; Chan, Helen; Verde, Ignazio; Main, Doreen; Jung, Sook; Georgi, Laura; Forrest, Sam; Mook, Jennifer; Zhebentyayeva, Tatyana; Yu, Yeisoo; Kim, Hye Ran; Jesudurai, Christopher; Sosinski, Bryon; Arús, Pere; Baird, Vance; Parfitt, Dan; Reighard, Gregory; Scorza, Ralph; Tomkins, Jeffrey; Wing, Rod; Abbott, Albert Glenn

    2005-05-01

    Peach (Prunus persica) is a model species for the Rosaceae, which includes a number of economically important fruit tree species. To develop an extensive Prunus expressed sequence tag (EST) database for identifying and cloning the genes important to fruit and tree development, we generated 9,984 high-quality ESTs from a peach cDNA library of developing fruit mesocarp. After assembly and annotation, a putative peach unigene set consisting of 3,842 ESTs was defined. Gene ontology (GO) classification was assigned based on the annotation of the single "best hit" match against the Swiss-Prot database. No significant homology could be found in the GenBank nr databases for 24.3% of the sequences. Using core markers from the general Prunus genetic map, we anchored bacterial artificial chromosome (BAC) clones on the genetic map, thereby providing a framework for the construction of a physical and transcript map. A transcript map was developed by hybridizing 1,236 ESTs from the putative peach unigene set and an additional 68 peach cDNA clones against the peach BAC library. Hybridizing ESTs to genetically anchored BACs immediately localized 11.2% of the ESTs on the genetic map. ESTs showed a clustering of expressed genes in defined regions of the linkage groups. [The data were built into a regularly updated Genome Database for Rosaceae (GDR), available at (http://www.genome.clemson.edu/gdr/).].

  18. Burdock fructooligosaccharide enhances biocontrol of Rhodotorula mucilaginosa to postharvest decay of peaches.

    Science.gov (United States)

    Zhang, Hongyin; Liu, Zhouyang; Xu, Baitian; Chen, Keping; Yang, Qiya; Zhang, Qiuyun

    2013-10-15

    The influence of adding burdock fructooligosaccharide (BFO) in the culture media on the efficacy of Rhodotorula mucilaginosa in controlling postharvest decay of peaches and its possible mode of action were investigated. The antagonistic activity of R. mucilaginosa to Rhizopus decay and blue mold decay of peaches was greatly enhanced through cultivation in the nutrient yeast dextrose agar (NYDA) medium amended with BFO at the concentration of 0.32%, compared with that cultivated in NYDB without BFO. R. mucilaginosa at 1×10(8) cells/mL cultivation in the NYDB media did not reduce the natural decay incidence of peaches, compared with the control after 30 d at 4 °C followed by 7d at 20 °C. However, R. mucilaginosa cultivation in the NYDB media amended with BFO at the concentration of 0.32% reduced the natural decay incidence of peaches. The population of R. mucilaginosa harvested from NYDB amended with BFO at 0.32% increased rapidly in peach wounds compared to that harvested from NYDB without BFO no matter peaches were stored at 20 °C or 4 °C. The activities of chitinase and β-1,3-glucanase of cell-free culture filtrate of R. mucilaginosa harvested from NYDB amended with BFO at 0.32% were higher than that at other concentrations and the control. Copyright © 2013 Elsevier Ltd. All rights reserved.

  19. Effect of gamma irradiation on refrigerator storage of peach fruits

    International Nuclear Information System (INIS)

    Kalinov, V.

    1985-01-01

    Peach fruits of cvs. Halle and Elberta were gamma irradiated by 0, 2.0, 2.5 and 3.0 kGy and stored in the usual way (20-25 grad C) or in refrigerator (5 and 0 grad C). The rate of mould decay, reduction in market quality and complex evaluation of the fruits at the time of storage were investigated. Linear regression correlations between the percentage of decay and the time of storage were estimated by probit-logarithmic transformation of the percentage of the decayed fruits. On that basis were assess the probable terms to reach 5% decay (permissible period of storage) and 10% decay. The criteria of quality and taste evaluation of the fruits for these periods were determined. The irradiation of peach fruits with 2.5 and 3.0 kGy prolonged 3 to 5 times the period free of decay (up to 5%) in the case of normal storage conditions as compared to the control (no irradiation). The sharp reduction in quality and the sensorial evaluation for these periods did not justify irradiation in case of usual storage conditions. Irradiation with the same rates in case of refrigeration storage proved a useful supplementary factor prolonging the periods before appearance of decay up to 17 days (i.e. 42% as compared to the control) and the permissible period of storage up to 25 days (25% more than the control). These results were obtained at a dose of 2.5 kGy for cv. Halle fruits, at dose 3.0 kGy for cv. Elberta fruits and at storage temperature of 0 grad C. For the periods up to 5% decay (25 to 26 days) the criteria of quality and the complex evaluation remained above the permissible level. Longer storage was not advisable because physiological damage of the fruit was evident

  20. Involvement of rho-gtpases in fibroblast adhesion and fibronectine fibrillogenesis under stretch

    Science.gov (United States)

    Guignandon, A.; Lambert, C.; Rattner, A.; Servotte, S.; Lapiere, C.; Nusgens, B.; Vico, L.

    The Rho family small GTPases play a crucial role in mediating cellular adaptation to mechanical stimulation (MS), and possibly to microgravity (μg), through effects on the cytoskeleton and cell adhesion which is, in turn, mainly regulated by fibronectin fibrillogenesis (FnF). It remains unclear how mechanical stimulation is transduced to the Rho signaling pathways and how it impacts on fibronectin (fbn) fibrillogenesis (FnF). μg (2 days, mission STS-095) led to de-adhesion of fibroblasts and modification of the underlying extracellular matrix. To determine whether GTPases modulated FnF, we generated stable cell lines expressing high level of activated RhoA and Rac1 (QL) as compared to wild type (WI26-WT). After MS application [8% deformation, 1Hz, 15 min., 3 times/day for 1-2 days], we quantified focal adhesion (vinculin, paxillin, FAKY397), f-actin stress fibers (Sf) and FnF with home-developed softwares. We reported that after MS, Sf are more rapidly (30min) formed under the nucleus in Wi26-WT (+100%) and Rac1 (+200%) than in RhoA (+20%). Vinculin & paxillin were only restricted to the cell edge in static conditions and homogeneously distributed after MS in WT and Rac1. The relative area of contacts (vinculin & paxillin) was more dramatically enhanced by MS in Rac1 (+80%) than in WT (+40%) and RhoA (+25%) indicating that new focal contacts are formed under MS and supported the presence of Sf. MS Activation of FAK (FAKY397) was clear in WT and Rac1 and reduced in RhoA. FnF was restricted to cell-cell contacts zone without any change in the relative area of fbn after a 2-days MS. However we found more numerous spots of fbn at the cell center in Rac1 as compared with RhoA & WT suggesting that these fibrillar contacts will grow upon maturation and modulate FnF. The results indicate that MS induces formation of Sf and focal adhesions and enhances FF. RhoA has been shown to induce the formation of Sf and focal adhesions, and Rac1 activation decreases Rho activity in

  1. Structural Dynamics Control Allosteric Activation of Cytohesin Family Arf GTPase Exchange Factors

    Energy Technology Data Exchange (ETDEWEB)

    Malaby, Andrew W.; Das, Sanchaita; Chakravarthy, Srinivas; Irving, Thomas C.; Bilsel, Osman; Lambright, David G.

    2018-01-01

    Membrane dynamic processes including vesicle biogenesis depend on Arf guanosine triphosphatase (GTPase) activation by guanine nucleotide exchange factors (GEFs) containing a catalytic Sec7 domain and a membrane-targeting module such as a pleckstrin homology (PH) domain. The catalytic output of cytohesin family Arf GEFs is controlled by autoinhibitory interactions that impede accessibility of the exchange site in the Sec7 domain. These restraints can be relieved through activator Arf-GTP binding to an allosteric site comprising the PH domain and proximal autoinhibitory elements (Sec7-PH linker and C-terminal helix). Small-angle X-ray scattering and negative-stain electron microscopy were used to investigate the structural organization and conformational dynamics of cytohesin-3 (Grp1) in autoinhibited and active states. The results support a model in which hinge dynamics in the autoinhibited state expose the activator site for Arf-GTP binding, while subsequent C-terminal helix unlatching and repositioning unleash conformational entropy in the Sec7-PH linker to drive exposure of the exchange site.

  2. Rho GTPases: Novel Players in the Regulation of the DNA Damage Response?

    Directory of Open Access Journals (Sweden)

    Gerhard Fritz

    2015-09-01

    Full Text Available The Ras-related C3 botulinum toxin substrate 1 (Rac1 belongs to the family of Ras-homologous small GTPases. It is well characterized as a membrane-bound signal transducing molecule that is involved in the regulation of cell motility and adhesion as well as cell cycle progression, mitosis, cell death and gene expression. Rac1 also adjusts cellular responses to genotoxic stress by regulating the activity of stress kinases, including c-Jun-N-terminal kinase/stress-activated protein kinase (JNK/SAPK and p38 kinases as well as related transcription factors. Apart from being found on the inner side of the outer cell membrane and in the cytosol, Rac1 has also been detected inside the nucleus. Different lines of evidence indicate that genotoxin-induced DNA damage is able to activate nuclear Rac1. The exact mechanisms involved and the biological consequences, however, are unclear. The data available so far indicate that Rac1 might integrate DNA damage independent and DNA damage dependent cellular stress responses following genotoxin treatment, thereby coordinating mechanisms of the DNA damage response (DDR that are related to DNA repair, survival and cell death.

  3. Rho-GTPase effector ROCK phosphorylates cofilin in actin-meditated cytokinesis during mouse oocyte meiosis.

    Science.gov (United States)

    Duan, Xing; Liu, Jun; Dai, Xiao-Xin; Liu, Hong-Lin; Cui, Xiang-Shun; Kim, Nam-Hyung; Wang, Zhen-Bo; Wang, Qiang; Sun, Shao-Chen

    2014-02-01

    During oocyte meiosis, a spindle forms in the central cytoplasm and migrates to the cortex. Subsequently, the oocyte extrudes a small body and forms a highly polarized egg; this process is regulated primarily by actin. ROCK is a Rho-GTPase effector that is involved in various cellular functions, such as stress fiber formation, cell migration, tumor cell invasion, and cell motility. In this study, we investigated possible roles for ROCK in mouse oocyte meiosis. ROCK was localized around spindles after germinal vesicle breakdown and was colocalized with cytoplasmic actin and mitochondria. Disrupting ROCK activity by RNAi or an inhibitor resulted in cell cycle progression and polar body extrusion failure. Time-lapse microscopy showed that this may have been due to spindle migration and cytokinesis defects, as chromosomes segregated but failed to extrude a polar body and then realigned. Actin expression at oocyte membranes and in cytoplasm was significantly decreased after these treatments. Actin caps were also disrupted, which was confirmed by a failure to form cortical granule-free domains. The mitochondrial distribution was also disrupted, which indicated that mitochondria were involved in the ROCK-mediated actin assembly. In addition, the phosphorylation levels of Cofilin, a downstream molecule of ROCK, decreased after disrupting ROCK activity. Thus, our results indicated that a ROCK-Cofilin-actin pathway regulated meiotic spindle migration and cytokinesis during mouse oocyte maturation.

  4. NMR derived model of GTPase effector domain (GED self association: relevance to dynamin assembly.

    Directory of Open Access Journals (Sweden)

    Swagata Chakraborty

    Full Text Available Self-association of dynamin to form spiral structures around lipidic vesicles during endocytosis is largely mediated by its 'coiled coil' GTPase Effector Domain (GED, which, in vitro, self-associates into huge helical assemblies. Residue-level structural characterizations of these assemblies and understanding the process of association have remained a challenge. It is also impossible to get folded monomers in the solution phase. In this context, we have developed here a strategy to probe the self-association of GED by first dissociating the assembly using Dimethyl Sulfoxide (DMSO and then systematically monitoring the refolding into helix and concomitant re-association using NMR spectroscopy, as DMSO concentration is progressively reduced. The short segment, Arg109 - Met116, acts as the nucleation site for helix formation and self-association. Hydrophobic and complementary charge interactions on the surfaces drive self-association, as the helices elongate in both the directions resulting in an antiparallel stack. A small N-terminal segment remains floppy in the assembly. Following these and other published results on inter-domain interactions, we have proposed a plausible mode of dynamin self assembly.

  5. The Rab2A GTPase Promotes Breast Cancer Stem Cells and Tumorigenesis via Erk Signaling Activation

    Directory of Open Access Journals (Sweden)

    Man-Li Luo

    2015-04-01

    Full Text Available Proline-directed phosphorylation is regulated by the prolyl isomerase Pin1, which plays a fundamental role in driving breast cancer stem-like cells (BCSCs. Rab2A is a small GTPase critical for vesicle trafficking. Here, we show that Pin1 increases Rab2A transcription to promote BCSC expansion and tumorigenesis in vitro and in vivo. Mechanistically, Rab2A directly interacts with and prevents dephosphorylation/inactivation of Erk1/2 by the MKP3 phosphatase, resulting in Zeb1 upregulation and β-catenin nuclear translocation. In cancer cells, Rab2A is activated via gene amplification, mutation or Pin1 overexpression. Rab2A overexpression or mutation endows BCSC traits to primary normal human breast epithelial cells, whereas silencing Rab2A potently inhibits the expansion and tumorigenesis of freshly isolated BCSCs. Finally, Rab2A overexpression correlates with poor clinical outcome in breast cancer patients. Thus, Pin1/Rab2A/Erk drives BCSC expansion and tumorigenicity, suggesting potential drug targets.

  6. GTPase ROP2 binds and promotes activation of target of rapamycin, TOR, in response to auxin.

    Science.gov (United States)

    Schepetilnikov, Mikhail; Makarian, Joelle; Srour, Ola; Geldreich, Angèle; Yang, Zhenbiao; Chicher, Johana; Hammann, Philippe; Ryabova, Lyubov A

    2017-04-03

    Target of rapamycin (TOR) promotes reinitiation at upstream ORFs (uORFs) in genes that play important roles in stem cell regulation and organogenesis in plants. Here, we report that the small GTPase ROP2, if activated by the phytohormone auxin, promotes activation of TOR, and thus translation reinitiation of uORF-containing mRNAs. Plants with high levels of active ROP2, including those expressing constitutively active ROP2 (CA-ROP2), contain high levels of active TOR ROP2 physically interacts with and, when GTP-bound, activates TOR in vitro TOR activation in response to auxin is abolished in ROP-deficient rop2 rop6 ROP4 RNAi plants. GFP-TOR can associate with endosome-like structures in ROP2-overexpressing plants, indicating that endosomes mediate ROP2 effects on TOR activation. CA-ROP2 is efficient in loading uORF-containing mRNAs onto polysomes and stimulates translation in protoplasts, and both processes are sensitive to TOR inhibitor AZD-8055. TOR inactivation abolishes ROP2 regulation of translation reinitiation, but not its effects on cytoskeleton or intracellular trafficking. These findings imply a mode of translation control whereby, as an upstream effector of TOR, ROP2 coordinates TOR function in translation reinitiation pathways in response to auxin. © 2017 The Authors.

  7. Interaction of LRRK2 with kinase and GTPase signaling cascades

    Directory of Open Access Journals (Sweden)

    Joon Y Boon

    2014-07-01

    Full Text Available LRRK2 is a protein that interacts with a plethora of signaling molecules, but the complexity of LRRK2 function presents a challenge for understanding the role of LRRK2 in the pathophysiology of Parkinson’s disease. Studies of LRRK2 using over-expression in transgenic mice have been disappointing, however studies using invertebrate systems have yielded a much clearer picture, with clear effects of LRRK2 expression, knockdown or deletion in C. elegans and Drosophila on modulation of survival of dopaminergic neurons. Recent studies have begun to focus attention on particular signaling cascades that are a target of LRRK2 function. LRRK2 interacts with members of the MAPK pathway and might regulate the pathway action by acting as a scaffold that directs the location of MAPK pathway activity, without strongly affecting the amount of MAPK pathway activity. Binding to GTPases, GAPs and GEFs are another strong theme in LRRK2 biology, with LRRK2 binding to Rac1, cdc42, rab5, rab7L1, endoA, RGS2, ArfGAP1 and ArhGEF7. All of these molecules appear to feed into a function output for LRRK2 that modulates cytoskeletal outgrowth and vesicular dynamics, including autophagy. These functions likely impact modulation of α-synuclein aggregation and associated toxicity eliciting the disease processes that we term Parkinson’s disease.

  8. Investigation into effects of ionizing radiations on physical-chemical properties of bulgarian sorts of peaches and grapes

    International Nuclear Information System (INIS)

    Dimitrov, D.; Kalinov, V.; Nikolova, M.; Pavlova, E.

    1974-01-01

    The aim of this study was to ascertain the degree and direction of those changes in physicochemical characteristics of irradiated peaches and grapes which determine their quality after irradiation and during storage. The following main conclusions are made: irradiation with doses of 200 to 300 krad does not cause significant alterations in the chemical composition and physicochemical characteristics of peaches and grapes; irradiation has a direct effect on the consistancy of peaches and grapes, leading to their softening which is proportional to the radiation dose used; radiation treatment is advisable in the case of peaches and grapes in tended for short-term rather than long-term storage at low temperatures

  9. [Genome-wide identification and expression analysis of the WRKY gene family in peach].

    Science.gov (United States)

    Gu, Yan-bing; Ji, Zhi-rui; Chi, Fu-mei; Qiao, Zhuang; Xu, Cheng-nan; Zhang, Jun-xiang; Zhou, Zong-shan; Dong, Qing-long

    2016-03-01

    The WRKY transcription factors are one of the largest families of transcriptional regulators and play diverse regulatory roles in biotic and abiotic stresses, plant growth and development processes. In this study, the WRKY DNA-binding domain (Pfam Database number: PF03106) downloaded from Pfam protein families database was exploited to identify WRKY genes from the peach (Prunus persica 'Lovell') genome using HMMER 3.0. The obtained amino acid sequences were analyzed with DNAMAN 5.0, WebLogo 3, MEGA 5.1, MapInspect and MEME bioinformatics softwares. Totally 61 peach WRKY genes were found in the peach genome. Our phylogenetic analysis revealed that peach WRKY genes were classified into three Groups: Ⅰ, Ⅱ and Ⅲ. The WRKY N-terminal and C-terminal domains of Group Ⅰ (group I-N and group I-C) were monophyletic. The Group Ⅱ was sub-divided into five distinct clades (groupⅡ-a, Ⅱ-b, Ⅱ-c, Ⅱ-d and Ⅱ-e). Our domain analysis indicated that the WRKY regions contained a highly conserved heptapeptide stretch WRKYGQK at its N-terminus followed by a zinc-finger motif. The chromosome mapping analysis showed that peach WRKY genes were distributed with different densities over 8 chromosomes. The intron-exon structure analysis revealed that structures of the WRKY gene were highly conserved in the peach. The conserved motif analysis showed that the conserved motifs 1, 2 and 3, which specify the WRKY domain, were observed in all peach WRKY proteins, motif 5 as the unknown domain was observed in group Ⅱ-d, two WRKY domains were assigned to GroupⅠ. SqRT-PCR and qRT-PCR results indicated that 16 PpWRKY genes were expressed in roots, stems, leaves, flowers and fruits at various expression levels. Our analysis thus identified the PpWRKY gene families, and future functional studies are needed to reveal its specific roles.

  10. Plant Rho-type (Rop) GTPase-dependent activation of receptor-like cytoplasmic kinases in vitro.

    Science.gov (United States)

    Dorjgotov, Dulguun; Jurca, Manuela E; Fodor-Dunai, Csilla; Szucs, Attila; Otvös, Krisztina; Klement, Eva; Bíró, Judit; Fehér, Attila

    2009-04-02

    Plants have evolved distinct mechanisms to link Rho-type (Rop) GTPases to downstream signaling pathways as compared to other eukaryotes. Here, experimental data are provided that members of the Medicago, as well as Arabidopsis, receptor-like cytoplasmic kinase family (RLCK Class VI) were strongly and specifically activated by GTP-bound Rop GTPases in vitro. Deletion analysis indicated that the residues implicated in the interaction might be distributed on various parts of the kinases. Using a chimaeric Rop GTPase protein, the importance of the Rho-insert region in kinase activation could also be verified. These data strengthen the possibility that RLCKs may serve as Rop GTPase effectors in planta.

  11. Human Lsg1 defines a family of essential GTPases that correlates with the evolution of compartmentalization

    Directory of Open Access Journals (Sweden)

    Scheffzek Klaus

    2005-10-01

    Full Text Available Abstract Background Compartmentalization is a key feature of eukaryotic cells, but its evolution remains poorly understood. GTPases are the oldest enzymes that use nucleotides as substrates and they participate in a wide range of cellular processes. Therefore, they are ideal tools for comparative genomic studies aimed at understanding how aspects of biological complexity such as cellular compartmentalization evolved. Results We describe the identification and characterization of a unique family of circularly permuted GTPases represented by the human orthologue of yeast Lsg1p. We placed the members of this family in the phylogenetic context of the YlqF Related GTPase (YRG family, which are present in Eukarya, Bacteria and Archea and include the stem cell regulator Nucleostemin. To extend the computational analysis, we showed that hLsg1 is an essential GTPase predominantly located in the endoplasmic reticulum and, in some cells, in Cajal bodies in the nucleus. Comparison of localization and siRNA datasets suggests that all members of the family are essential GTPases that have increased in number as the compartmentalization of the eukaryotic cell and the ribosome biogenesis pathway have evolved. Conclusion We propose a scenario, consistent with our data, for the evolution of this family: cytoplasmic components were first acquired, followed by nuclear components, and finally the mitochondrial and chloroplast elements were derived from different bacterial species, in parallel with the formation of the nucleolus and the specialization of nuclear components.

  12. A remarkable synergistic effect at the transcriptomic level in peach fruits doubly infected by prunus necrotic ringspot virus and peach latent mosaic viroid.

    Science.gov (United States)

    Herranz, Mari Carmen; Niehl, Annette; Rosales, Marlene; Fiore, Nicola; Zamorano, Alan; Granell, Antonio; Pallas, Vicente

    2013-05-28

    Microarray profiling is a powerful technique to investigate expression changes of large amounts of genes in response to specific environmental conditions. The majority of the studies investigating gene expression changes in virus-infected plants are limited to interactions between a virus and a model host plant, which usually is Arabidopsis thaliana or Nicotiana benthamiana. In the present work, we performed microarray profiling to explore changes in the expression profile of field-grown Prunus persica (peach) originating from Chile upon single and double infection with Prunus necrotic ringspot virus (PNRSV) and Peach latent mosaic viroid (PLMVd), worldwide natural pathogens of peach trees. Upon single PLMVd or PNRSV infection, the number of statistically significant gene expression changes was relatively low. By contrast, doubly-infected fruits presented a high number of differentially regulated genes. Among these, down-regulated genes were prevalent. Functional categorization of the gene expression changes upon double PLMVd and PNRSV infection revealed protein modification and degradation as the functional category with the highest percentage of repressed genes whereas induced genes encoded mainly proteins related to phosphate, C-compound and carbohydrate metabolism and also protein modification. Overrepresentation analysis upon double infection with PLMVd and PNRSV revealed specific functional categories over- and underrepresented among the repressed genes indicating active counter-defense mechanisms of the pathogens during infection. Our results identify a novel synergistic effect of PLMVd and PNRSV on the transcriptome of peach fruits. We demonstrate that mixed infections, which occur frequently in field conditions, result in a more complex transcriptional response than that observed in single infections. Thus, our data demonstrate for the first time that the simultaneous infection of a viroid and a plant virus synergistically affect the host transcriptome in

  13. Carbon Monoxide Fumigation Improved the Quality, Nutrients, and Antioxidant Activities of Postharvest Peach

    Science.gov (United States)

    Li, Ying; Pei, Fei

    2014-01-01

    Peaches (Prunus persica cv. Yanhong) were fumigated with carbon monoxide (CO) at 0, 0.5, 5, 10, and 20 μmol/L for 2 hours. The result showed that low concentration CO (0.5–10 μmol/L) might delay the decrease of firmness and titrable acid content, restrain the increase of decay incidence, and postpone the variation of soluble solids content, but treating peaches with high concentration CO (20 μmol/L) demonstrated adverse effects. Further research exhibited that exogenous CO could induce the phenylalnine ammonialyase activity, maintain nutrient contents such as Vitamin C, total flavonoid, and polyphenol, and enhance antioxidant activity according to reducing power and 2,2-diphenyl-1-(2,4,6-trinitrophenyl) hydrazyl radical scavenging activity. Treating peaches with appropriate concentration CO was beneficial to the quality, nutrients, and antioxidant activity of postharvest peaches during storage time. Therefore, CO fumigation might probably become a novel method to preserve postharvest peach and other fruits in the future. PMID:26904651

  14. Intake, digestibility and performance of lambs fed diets containing peach palm meal.

    Science.gov (United States)

    dos Santos, Alana Batista; Pereira, Mara Lúcia Albuquerque; de Oliveira Silva, Herymá Giovane; de Carvalho, Gleidson Giordano Pinto; de Jesus Pereira, Taiala Cristina; Ribeiro, Leandro Sampaio Oliveira; Azevêdo, José Augusto Gomes; das Graças Conceição Parada Costa Silva, Maria; Sousa, Larisse Borges; Sousa, Leandro Borges; de Oliveira Alencar, Daiane

    2016-03-01

    The aim of this study was to evaluate the intake and apparent digestibility of nutrients, performance, and plasma glucose concentration of ram lambs fed diets containing peach palm meal substituting maize (0, 10, 40, 60, and 85 % dry matter (DM)). Thirty Santa Inês rams with an average initial body weight of 21.6 ± 0.87 kg were distributed in a completely randomized design with five diets and six replicates. The substitution of the maize for the peach palm meal affected (P < 0.05) the intakes of DM, organic matter (OM), crude protein (CP), neutral detergent fiber corrected for ash and protein (NDFap), total carbohydrates (TC), total digestible nutrients (TDN), and metabolizable energy (ME), which decreased linearly (P < 0.05); the intake of ether extract (EE), however, fit an increasing linear equation (P < 0.05). The apparent digestibility coefficients of DM, OM, NDFap, and TC decreased linearly (P < 0.05) as the level of peach palm meal in the concentrate was increased. The total weight gain and the average daily gain decreased by 0.09 and 0.001 kg with each level of substitution of the maize for peach palm meal, respectively. It is recommended to substitute 40 % of the maize for peach palm meal.

  15. Genetic analysis identifies the region of origin of smuggled peach palm seeds.

    Science.gov (United States)

    Cristo-Araújo, Michelly; Molles, David Bronze; Rodrigues, Doriane Picanço; Clement, Charles R

    2017-04-01

    Seeds of a plant, supposedly a palm tree known popularly as peach palm (Bactris gasipaes), were seized by the Federal Police in the state of Pará, Brazil, without documentation of legal origin to authorize transportation and marketing in Brazil. They were alleged to be from the western part of Amazonas, Brazil, near the frontier with Peru and Colombia, justifying the lack of documentation. The species was confirmed to be peach palm. To determine the likely place of origin, a genetic analysis was performed to determine the relationship between the seized seeds and representative populations of peach palm from all of Amazonia, maintained in the Peach palm Core Collection, at the National Research Institute for Amazonia, using nine microsatellite loci. Reynolds' coancestry analysis showed a strong relationship between the seeds and the Pampa Hermosa landrace, around Yurimaguas, Peru. The Structure program, used to infer the probability of an individual belonging to a given population, showed that most seeds grouped with populations close to Yurimaguas, Peru, corroborating the coancestry analysis. The Pampa Hermosa landrace is the main source of spineless peach palm seeds used in the Brazilian heart-of-palm agribusiness, which motivated the smugglers to attempt this biopiracy. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Decolorization of textile industry wastewater in solid state fermentation with Peach-Palm (Bactris gasipaes) residue.

    Science.gov (United States)

    Chicatto, J A; Rainert, K T; Gonçalves, M J; Helm, C V; Altmajer-Vaz, D; Tavares, L B B

    2018-02-15

    In this work we have assessed the decolorization of textile effluents throughout their treatment in a solid-state fermentation (SSF) system. SSF assays were conducted with peach-palm (Bactris gasipaes) residue using the white rot fungus Ganoderma lucidum EF 31. The influence of the dye concentration and of the amounts of peach-palm residue and liquid phase on both the discoloration efficiency and enzyme production was studied. According to our results, independently of experimental conditions employed, laccase was the main ligninolytic enzyme produced by G. lucidum. The highest laccase activity was obtained at very low effluent concentrations, suggesting the existence of an inhibitory effect of higher concentrations on fungal metabolism. The highest percentage of color removal was reached when 10 grams of peach palm residue was moistened with 60 mL of the final effluent. In control tests carried out with the synthetic dye Remazol Brilliant Blue R (RBBR) decolorization efficiencies about 20% higher than that achieved with the industrial effluent were achieved. The adsorption of RBBR on peach-palm residue was also investigated. Equilibrium tests showed that the adsorption of this dye followed both Langmuir and Freundlich isotherms. Hence, our experimental results indicate that peach-palm residue is suitable substrate for both laccase production and color removal in industrial effluents.

  17. Decolorization of textile industry wastewater in solid state fermentation with Peach-Palm (Bactris gasipaes residue

    Directory of Open Access Journals (Sweden)

    J. A. Chicatto

    2018-02-01

    Full Text Available Abstract In this work we have assessed the decolorization of textile effluents throughout their treatment in a solid-state fermentation (SSF system. SSF assays were conducted with peach-palm (Bactris gasipaes residue using the white rot fungus Ganoderma lucidum EF 31. The influence of the dye concentration and of the amounts of peach-palm residue and liquid phase on both the discoloration efficiency and enzyme production was studied. According to our results, independently of experimental conditions employed, laccase was the main ligninolytic enzyme produced by G. lucidum. The highest laccase activity was obtained at very low effluent concentrations, suggesting the existence of an inhibitory effect of higher concentrations on fungal metabolism. The highest percentage of color removal was reached when 10 grams of peach palm residue was moistened with 60 mL of the final effluent. In control tests carried out with the synthetic dye Remazol Brilliant Blue R (RBBR decolorization efficiencies about 20% higher than that achieved with the industrial effluent were achieved. The adsorption of RBBR on peach-palm residue was also investigated. Equilibrium tests showed that the adsorption of this dye followed both Langmuir and Freundlich isotherms. Hence, our experimental results indicate that peach-palm residue is suitable substrate for both laccase production and color removal in industrial effluents.

  18. Differential metabolic rearrangements after cold storage are correlated with chilling injury resistance of peach fruits

    Directory of Open Access Journals (Sweden)

    Claudia A Bustamante

    2016-09-01

    Full Text Available Reconfiguration of the metabolome is a key component involved in the acclimation to cold in plants; however, few studies have been devoted to the analysis of the overall metabolite changes after cold storage of fruits prior to consumption. Here, metabolite profiling of six peach varieties with differential susceptibility to develop mealiness, a chilling-injury (CI symptom, was performed. According to metabolic content at harvest; after cold treatment; and after ripening, either following cold treatment or not; peach fruits clustered in distinct groups, depending on harvest-time, cold treatment, and ripening state. Both common and distinct metabolic responses among the six varieties were found; common changes including dramatic galactinol and raffinose rise; GABA, Asp and Phe increase; and 2-oxo-glutarate and succinate decrease. Raffinose content after long cold treatment quantitatively correlated to the degree of mealiness resistance of the different peach varieties; and thus, raffinose emerges as a candidate biomarker of this CI disorder. Xylose increase after cold treatment was found only in the susceptible genotypes, indicating a particular cell wall reconfiguration of these varieties while being cold-stored. Overall, results indicate that peach fruit differential metabolic rearrangements due to cold treatment, rather than differential metabolic priming before cold, are better related with CI resistance. The plasticity of peach fruit metabolism renders it possible to induce a diverse metabolite array after cold, which is successful, in some genotypes, to avoid CI

  19. Phenotypic characterization of qualitative parameters and antioxidant contents in peach and nectarine fruit and changes after jam preparation.

    Science.gov (United States)

    Drogoudi, Pavlina; Gerasopoulos, Dimitrios; Kafkaletou, Mina; Tsantili, Eleni

    2017-08-01

    Sugars and antioxidants in peaches contribute to fresh fruit quality and nutrition; however, information on widely grown cultivars and changes induced after peach jam preparation is limited. In the present study, colour, sugars and antioxidant parameters were determined in fruit and jam from 45 peach and nectarine cultivars. Pronounced varietal differences were found in sorbitol (42-fold range), total phenolics (TPs) and antioxidant capacities (10- to 19-fold range). Sorbitol levels were greater in non-melting peach, followed by nectarine, and lower values were found in melting peach cultivars. Late-harvested peach and nectarine cultivars tended to have a higher soluble solid content and antioxidant potential. Cultivars with relatively high antioxidant contents produced darker and redder jams, containing more antioxidants, than the jam or the fruit from the other cultivars. Jam-TPs were reduced by 48% compared to fruit-TPs, with greater reduction being noted in high antioxidant cultivars. The most favorable jam organoleptic characteristics were found in 'Morsiani 90', 'Amiga', 'Romea' and 'Alirosada', as well as in non-melting compared to melting peach cultivars. The best cultivars for each fruit flesh type and jam were identified. Peach jam could be an alternative substitute when fresh fruit is not available and when it is prepared with high antioxidant cultivars. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  20. Peach leaf responses to soil and cement dust pollution.

    Science.gov (United States)

    Maletsika, Persefoni A; Nanos, George D; Stavroulakis, George G

    2015-10-01

    Dust pollution can negatively affect plant productivity in hot, dry and with high irradiance areas during summer. Soil or cement dust were applied on peach trees growing in a Mediterranean area with the above climatic characteristics. Soil and cement dust accumulation onto the leaves decreased the photosynthetically active radiation (PAR) available to the leaves without causing any shade effect. Soil and mainly cement dust deposition onto the leaves decreased stomatal conductance, photosynthetic and transpiration rates, and water use efficiency due possibly to stomatal blockage and other leaf cellular effects. In early autumn, rain events removed soil dust and leaf functions partly recovered, while cement dust created a crust partially remaining onto the leaves and causing more permanent stress. Leaf characteristics were differentially affected by the two dusts studied due to their different hydraulic properties. Leaf total chlorophyll decreased and total phenol content increased with dust accumulation late in the summer compared to control leaves due to intense oxidative stress. The two dusts did not cause serious metal imbalances to the leaves, except of lower leaf K content.

  1. Sensitivity analyses of the peach bottom turbine trip 2 experiment

    International Nuclear Information System (INIS)

    Bousbia Salah, A.; D'Auria, F.

    2003-01-01

    In the light of the sustained development in computer technology, the possibilities for code calculations in predicting more realistic transient scenarios in nuclear power plants have been enlarged substantially. Therefore, it becomes feasible to perform 'Best-estimate' simulations through the incorporation of three-dimensional modeling of reactor core into system codes. This method is particularly suited for complex transients that involve strong feedback effects between thermal-hydraulics and kinetics as well as to transient involving local asymmetric effects. The Peach bottom turbine trip test is characterized by a prompt core power excursion followed by a self limiting power behavior. To emphasize and understand the feedback mechanisms involved during this transient, a series of sensitivity analyses were carried out. This should allow the characterization of discrepancies between measured and calculated trends and assess the impact of the thermal-hydraulic and kinetic response of the used models. On the whole, the data comparison revealed a close dependency of the power excursion with the core feedback mechanisms. Thus for a better best estimate simulation of the transient, both of the thermal-hydraulic and the kinetic models should be made more accurate. (author)

  2. Peach bottom cycle 2 stability analysis using RELAP5/PARCS

    International Nuclear Information System (INIS)

    Maggini, F.; D'Auria, F.; Miro, R.; Verdu, G.; Ginestar, D.

    2003-01-01

    Boiling channels and systems may oscillate owing to the behaviour of the liquid-steam mixture used for removing the thermal power. A thermal-hydraulic system may be unstable under particular operating conditions. Two kinds of power oscillation have been observed in BWR cores. One is an in-phase (core-wide) and the other is an out-of-phase (regional) oscillation. Since the above feature can make detection more difficult, the latter oscillation is potentially more severe. The problem is well known since the design of the first BWR system. However, to improve the safety systems of these reactors, it is necessary to be able to detect in a reliable way these oscillations from the neutronic signals. The purpose of this work is to characterize the unstable behaviour of a BWR. Within this study, it has been performed a number of perturbation analysis. The coupled codes RELAP5-Mod3.3/PARCS have used for the simulation of the transients. Validation has been performed against Peach Bottom-2 Low-Flow Stability Test PT3. Three dimensional time domain BWR stability analysis were performed on test point 3 for the core wide oscillation mode. In this transient dynamically complex events take place, i.e., neutron kinetics is coupled with thermal-hydraulics and an in-phase oscillation has been developed. The calculated results are compared against the available experimental data. (author)

  3. SENTINEL trademark technical basis report for Peach Bottom. Final report

    International Nuclear Information System (INIS)

    1998-04-01

    PECO Energy in cooperation with the Electric Power Research Institute (EPRI) installed the SENTINEL trademark software at its Peach Bottom Atomic Power Station (PBAPS). This software incorporates models of the safety and support systems which are used to display the defense in depth present in the plant and a quantitative assessment of the plant risks during proposed on-line maintenance. During the past nine months, PECO Energy personnel have used this display to evaluate the safety of proposed on-line maintenance schedules. The report describes the motivation for and the development of the SENTINEL software. It describes the generation of Safety Function Assessment Trees and Plant Transient Assessment Trees and their use in evaluating the level of defense-in-depth of key plant safety functions and the susceptibility of the plant to critical transient events. Their results are displayed by color indicators ranging from green, through yellow and orange, to red to show increasingly hazardous conditions. The report describes the use of the PBAPS Probabilistic Safety Assessment within the SENTINEL code to calculate an instantaneous core damage frequency and the criteria by which this frequency is translated to a color indicator

  4. Irradiation experience with HTGR fuels in the Peach Bottom Reactor

    International Nuclear Information System (INIS)

    Scheffel, W.J.; Scott, C.B.

    1974-01-01

    Fuel performance in the Peach Bottom High-Temperature Gas-Cooled Reactor (HTGR) is reviewed, including (1) the driver elements in the second core and (2) the test elements designed to test fuel for larger HTGR plants. Core 2 of this reactor, which is operated by the Philadelphia Electric Company, performed reliably with an average nuclear steam supply availability of 85 percent since its startup in July 1970. Core 2 had accumulated a total of 897.5 equivalent full power days (EFPD), almost exactly its design life-time of 900 EFPD, when the plant was shut down permanently on October 31, 1974. Gaseous fission product release and the activity of the main circulating loop remained significantly below the limits allowed by the technical specifications and the levels observed during operation of Core 1. The low circulating activity and postirradiation examination of driver fuel elements have demonstrated the improved irradiation stability of the coated fuel particles in Core 2. Irradiation data obtained from these tests substantiate the performance predictions based on accelerated tests and complement the fuel design effort by providing irradiation data in the low neutron fluence region

  5. Fission product behaviour in the Peach Bottom and Fort St. Vrain HTGRs

    International Nuclear Information System (INIS)

    Hanson, D.L.; Baldwin, N.L.; Strong, D.E.

    1981-01-01

    Actual operating data from the Peach Bottom (PB) and Fort St. Vrain (FSV) High-Temperature Gas-Cooled Reactors (HTGRs) have been compared with code predictions to assess the validity of the methods used to predict the behaviour of fission products in the primary coolant circuit. For both reactors the measured circuit activities were significantly below design values, and the observations generally verify the codes used for large HTGR design. The PB primary circuit after seven years of operation was exceptionally clean. A fuel element purge system virtually eliminated the release of fission gases into the primary coolant circuit. Extensive examinations at end-of-life revealed that only Cs and trace amounts of Sr had plated out in the circuit. Their plateout distributions were in excellent agreement with PAD code predictions. Most of the deposited activity was associated with carbonaceous surface films which resulted from occasional small inleakages of lubricating oil. Primary circuit activities in FSV during the first cycle were also very low. Noble gas activity was about 1% of the design limit; and the circulating iodines were at least one order of magnitude below the limit, although the measurement uncertainties are significant. The plateout per pass of the iodine isotopes increased with decreasing half-life (the value for I-131 is about 1% per pass) as predicted with the PADLOC code. Gamma scanning of two helium circulators indicated very low plateout activities. Iodine-131 was the principal fission product observed, along with small amounts of Cs-134, Cs-137, and Ba/La-140. (author)

  6. Uncoupling of dynamin polymerization and GTPase activity revealed by the conformation-specific nanobody dynab.

    Science.gov (United States)

    Galli, Valentina; Sebastian, Rafael; Moutel, Sandrine; Ecard, Jason; Perez, Franck; Roux, Aurélien

    2017-10-12

    Dynamin is a large GTPase that forms a helical collar at the neck of endocytic pits, and catalyzes membrane fission (Schmid and Frolov, 2011; Ferguson and De Camilli, 2012). Dynamin fission reaction is strictly dependent on GTP hydrolysis, but how fission is mediated is still debated (Antonny et al., 2016): GTP energy could be spent in membrane constriction required for fission, or in disassembly of the dynamin polymer to trigger fission. To follow dynamin GTP hydrolysis at endocytic pits, we generated a conformation-specific nanobody called dynab, that binds preferentially to the GTP hydrolytic state of dynamin-1. Dynab allowed us to follow the GTPase activity of dynamin-1 in real-time. We show that in fibroblasts, dynamin GTP hydrolysis occurs as stochastic bursts, which are randomly distributed relatively to the peak of dynamin assembly. Thus, dynamin disassembly is not coupled to GTPase activity, supporting that the GTP energy is primarily spent in constriction.

  7. Characterization of Amazon fibers of the peach palm, balsa, and babassu by XDR, TGA and NMR

    International Nuclear Information System (INIS)

    Martins, Maria A.; Marconcini, Jose M.; Morelli, Carolina L.; Marinelli, Alessandra L.; Bretas, Rosario E.S.

    2011-01-01

    The aim of this work was to present the results by testing X-ray diffraction (XRD), thermogravimetric analysis (TG), nuclear magnetic resonance (NMR) and determining the moisture content of the peach palm, balsa and babassu fibers for assessing the feasibility of composite materials. The fibers of peach palm, balsa and babassu showed characteristic chemical structure of lignocellulosic material, and good thermal stability up to 220 deg C. The fiber with the highest crystallinity index (Ic) is the peach palm (72%) and the less crystalline is the babassu (37%), while the balsa fibers have Ic equal to 64%. The results have shown that these fibers can be used in the manufacture of composite materials. (author)

  8. The effect of ultraviolet irradiation on shelf-life and ripening of peaches and apples

    International Nuclear Information System (INIS)

    Lu, J.Y.; Stevens, C.; Khan, V.A.; Kabwe, M.; Wilson, C.L.

    1991-01-01

    ''Loring'' and ''Elberta'' peaches and ''Golden Delicious'' apples were irradiated with UV (254nm) to doses of 0.84 X 10(4) to 40 X 10(4) erg/mm2 then stored. ''Loring'' were stored 10 days and ''Elberta'' 20 days at 12 degrees C. ''Golden Delicious'' were stored 30 days at 20-25 degrees C in a dark room. Fruit were examined and differences in percentage rot and in physical-chemical properties determined. Percentage rot decreased with increasing UV dose. Fruit were firmer, pH and soluble solids lower and acidity higher for UV-treated than for nontreated peaches; pH was lower and acidity and ascorbic acid higher in UV treated than in nontreated apples. Percentage weight loss was less for UV-treated apples. The results indicated that UV treatment not only reduced storage rots but also delayed ripening of peaches and apples

  9. The Synthesis and Characterization of Peach-Like ZnO

    International Nuclear Information System (INIS)

    Kamalianfar, A.; Halim, S. A.; Navasery, M.; Din Fasih, Ud; Lim, K. P.; Chen, S. K.; Jahromi, Siamak Pilban; Zahedi, J. A. M.

    2012-01-01

    Peach-like ZnO microstructures are synthesized using vapor phase transport on MgO (001) substrates with a copper oxide (60 nm) buffer layer. The structure and morphology of the product are investigated using an x-ray diffractometer (XRD) and a field-emission scanning electron microscope. The peaches have an average diameter of 3 μm and a wurtzite structure. To study the optical properties, photoluminescence (PL) and Raman spectroscopy are employed. A strong UV emission at 380 nm in the PL spectra is observed, and a sharp and dominant peak at 437 cm −1 in the Raman spectrum can be assigned to the good crystallization of obtained product. In addition, the growth mechanism of the peach-like ZnO structure is tentatively investigated based on the EDX analysis and growth time

  10. Palmitoylation of the immunity related GTPase, Irgm1: impact on membrane localization and ability to promote mitochondrial fission.

    Directory of Open Access Journals (Sweden)

    Stanley C Henry

    Full Text Available The Immunity-Related GTPases (IRG are a family of large GTPases that mediate innate immune responses. Irgm1 is particularly critical for immunity to bacteria and protozoa, and for inflammatory homeostasis in the intestine. Although precise functions for Irgm1 have not been identified, prior studies have suggested roles in autophagy/mitophagy, phagosome remodeling, cell motility, and regulating the activity of other IRG proteins. These functions ostensibly hinge on the ability of Irgm1 to localize to intracellular membranes, such as those of the Golgi apparatus and mitochondria. Previously, it has been shown that an amphipathic helix, the αK helix, in the C-terminal portion of the protein partially mediates membrane binding. However, in absence of αK, there is still substantial binding of Irgm1 to cellular membranes, suggesting the presence of other membrane binding motifs. In the current work, an additional membrane localization motif was found in the form of palmitoylation at a cluster of cysteines near the αK. An Irgm1 mutant possessing alanine to cysteine substitutions at these amino acids demonstrated little residual palmitoylation, yet it displayed only a small decrease in localization to the Golgi and mitochondria. In contrast, a mutant containing the palmitoylation mutations in combination with mutations disrupting the amphipathic character of the αK displayed a complete loss of apparent localization to the Golgi and mitochondria, as well as an overall loss of association with cellular membranes in general. Additionally, Irgm1 was found to promote mitochondrial fission, and this function was undermined in Irgm1 mutants lacking the palmitoylation domain, and to a greater extent in those lacking the αK, or the αK and palmitoylation domains combined. Our data suggest that palmitoylation together with the αK helix firmly anchor Irgm1 in the Golgi and mitochondria, thus facilitating function of the protein.

  11. CONTRIBUTIONS TO IMPROVING CULTURE TEHNOLOGIES OF PEACHES GROWN ON SANDY SOILS THE SOUTH OF OLTENIA

    Directory of Open Access Journals (Sweden)

    Anica Durau

    2013-12-01

    Full Text Available Technological factors with major implications in obtaining high yields and quality in peaches grown on sandy soils are planting row distance and shape of the crown, soil maintenance system, chemical, organic and foliar fertilzation. A smal size combined with the flatening of the crowns of the tres alows a dense planting, also ensure proper mechanization of work and easy penetration of light to the leaves and fruits. Crown form vertical belt proved to be suitable for al planting distances studied, easily made and maintained, having fruit production ranged betwen 15.9 t / ha at a distance of 2 m, 10.3 t / ha at a distance of 2.5 m and 7.9 t / ha at a distance of 3 m. The state of soil nutrient supply influence sucesful peach crop on sandy soils. The fertilzer dose of technology to N10 P80 K10 kg s.a / ha production was 34.9 t / ha. Organic fertilzation also contributes to obtaining high yields of peach. In sandy soil conditions most fruit production of 9.6 t / ha was obtained by fertilzation with organic manure 60t/ha. Besides fertilzation, soil maintenance system is one important link in the technology peach crop on sandy soils. The results found that the biggest peach fruit production was obtained from field maintenance system black-8,2t/ha. Using technology in foliar peaches culture on sandy soils, is an important means of providing nutrients that lead to improved proceses of growth and fructification. The best way is with foliar fertilzation Folibor in dose 5l/ha, the production obtained was 12.4 t /ha.

  12. Rho GTPase protein Cdc42 is critical for postnatal cartilage development

    International Nuclear Information System (INIS)

    Nagahama, Ryo; Yamada, Atsushi; Tanaka, Junichi; Aizawa, Ryo; Suzuki, Dai; Kassai, Hidetoshi; Yamamoto, Matsuo; Mishima, Kenji; Aiba, Atsu; Maki, Koutaro; Kamijo, Ryutaro

    2016-01-01

    Cdc42, a small Rho GTPase family member, has been shown to regulate multiple cellular functions in vitro, including actin cytoskeletal reorganization, cell migration, proliferation, and gene expression. However, its tissue-specific roles in vivo remain largely unknown, especially in postnatal cartilage development, as cartilage-specific Cdc42 inactivated mice die within a few days after birth. In this study, we investigated the physiological functions of Cdc42 during cartilage development after birth using tamoxifen-induced cartilage-specific inactivated Cdc42 conditional knockout (Cdc42 "f"l"/"f"l; Col2-CreERT) mice, which were generated by crossing Cdc42 flox mice (Cdc42 "f"l"/"f"l) with tamoxifen-induced type II collagen (Col2) Cre transgenic mice using a Cre/loxP system. The gross morphology of the Cdc42 cKO mice was shorter limbs and body, as well as reduced body weight as compared with the controls. In addition, severe defects were found in growth plate chondrocytes of the long bones, characterized by a shorter proliferating zone (PZ), wider hypertrophic zone (HZ), and loss of columnar organization of proliferating chondrocytes, resulting in delayed endochondral bone formation associated with abnormal bone growth. Our findings demonstrate the importance of Cdc42 for cartilage development during both embryonic and postnatal stages. - Highlights: • Tamoxifen-induced cartilage specific inactivated Cdc42 mutant mice were generated. • Cdc42 mutant mice were shorter limbs and body. • Severe defects were found in growth plate chondrocytes.

  13. Activation of Ran GTPase by a Legionella effector promotes microtubule polymerization, pathogen vacuole motility and infection.

    Directory of Open Access Journals (Sweden)

    Eva Rothmeier

    2013-09-01

    Full Text Available The causative agent of Legionnaires' disease, Legionella pneumophila, uses the Icm/Dot type IV secretion system (T4SS to form in phagocytes a distinct "Legionella-containing vacuole" (LCV, which intercepts endosomal and secretory vesicle trafficking. Proteomics revealed the presence of the small GTPase Ran and its effector RanBP1 on purified LCVs. Here we validate that Ran and RanBP1 localize to LCVs and promote intracellular growth of L. pneumophila. Moreover, the L. pneumophila protein LegG1, which contains putative RCC1 Ran guanine nucleotide exchange factor (GEF domains, accumulates on LCVs in an Icm/Dot-dependent manner. L. pneumophila wild-type bacteria, but not strains lacking LegG1 or a functional Icm/Dot T4SS, activate Ran on LCVs, while purified LegG1 produces active Ran(GTP in cell lysates. L. pneumophila lacking legG1 is compromised for intracellular growth in macrophages and amoebae, yet is as cytotoxic as the wild-type strain. A downstream effect of LegG1 is to stabilize microtubules, as revealed by conventional and stimulated emission depletion (STED fluorescence microscopy, subcellular fractionation and Western blot, or by microbial microinjection through the T3SS of a Yersinia strain lacking endogenous effectors. Real-time fluorescence imaging indicates that LCVs harboring wild-type L. pneumophila rapidly move along microtubules, while LCVs harboring ΔlegG1 mutant bacteria are stalled. Together, our results demonstrate that Ran activation and RanBP1 promote LCV formation, and the Icm/Dot substrate LegG1 functions as a bacterial Ran activator, which localizes to LCVs and promotes microtubule stabilization, LCV motility as well as intracellular replication of L. pneumophila.

  14. Activation of Ran GTPase by a Legionella Effector Promotes Microtubule Polymerization, Pathogen Vacuole Motility and Infection

    Science.gov (United States)

    Rothmeier, Eva; Pfaffinger, Gudrun; Hoffmann, Christine; Harrison, Christopher F.; Grabmayr, Heinrich; Repnik, Urska; Hannemann, Mandy; Wölke, Stefan; Bausch, Andreas; Griffiths, Gareth; Müller-Taubenberger, Annette; Itzen, Aymelt; Hilbi, Hubert

    2013-01-01

    The causative agent of Legionnaires' disease, Legionella pneumophila, uses the Icm/Dot type IV secretion system (T4SS) to form in phagocytes a distinct “Legionella-containing vacuole” (LCV), which intercepts endosomal and secretory vesicle trafficking. Proteomics revealed the presence of the small GTPase Ran and its effector RanBP1 on purified LCVs. Here we validate that Ran and RanBP1 localize to LCVs and promote intracellular growth of L. pneumophila. Moreover, the L. pneumophila protein LegG1, which contains putative RCC1 Ran guanine nucleotide exchange factor (GEF) domains, accumulates on LCVs in an Icm/Dot-dependent manner. L. pneumophila wild-type bacteria, but not strains lacking LegG1 or a functional Icm/Dot T4SS, activate Ran on LCVs, while purified LegG1 produces active Ran(GTP) in cell lysates. L. pneumophila lacking legG1 is compromised for intracellular growth in macrophages and amoebae, yet is as cytotoxic as the wild-type strain. A downstream effect of LegG1 is to stabilize microtubules, as revealed by conventional and stimulated emission depletion (STED) fluorescence microscopy, subcellular fractionation and Western blot, or by microbial microinjection through the T3SS of a Yersinia strain lacking endogenous effectors. Real-time fluorescence imaging indicates that LCVs harboring wild-type L. pneumophila rapidly move along microtubules, while LCVs harboring ΔlegG1 mutant bacteria are stalled. Together, our results demonstrate that Ran activation and RanBP1 promote LCV formation, and the Icm/Dot substrate LegG1 functions as a bacterial Ran activator, which localizes to LCVs and promotes microtubule stabilization, LCV motility as well as intracellular replication of L. pneumophila. PMID:24068924

  15. Amphetamine activates Rho GTPase signaling to mediate dopamine transporter internalization and acute behavioral effects of amphetamine

    Science.gov (United States)

    Wheeler, David S.; Underhill, Suzanne M.; Stolz, Donna B.; Murdoch, Geoffrey H.; Thiels, Edda; Romero, Guillermo; Amara, Susan G.

    2015-01-01

    Acute amphetamine (AMPH) exposure elevates extracellular dopamine through a variety of mechanisms that include inhibition of dopamine reuptake, depletion of vesicular stores, and facilitation of dopamine efflux across the plasma membrane. Recent work has shown that the DAT substrate AMPH, unlike cocaine and other nontransported blockers, can also stimulate endocytosis of the plasma membrane dopamine transporter (DAT). Here, we show that when AMPH enters the cytoplasm it rapidly stimulates DAT internalization through a dynamin-dependent, clathrin-independent process. This effect, which can be observed in transfected cells, cultured dopamine neurons, and midbrain slices, is mediated by activation of the small GTPase RhoA. Inhibition of RhoA activity with C3 exotoxin or a dominant-negative RhoA blocks AMPH-induced DAT internalization. These actions depend on AMPH entry into the cell and are blocked by the DAT inhibitor cocaine. AMPH also stimulates cAMP accumulation and PKA-dependent inactivation of RhoA, thus providing a mechanism whereby PKA- and RhoA-dependent signaling pathways can interact to regulate the timing and robustness of AMPH’s effects on DAT internalization. Consistent with this model, the activation of D1/D5 receptors that couple to PKA in dopamine neurons antagonizes RhoA activation, DAT internalization, and hyperlocomotion observed in mice after AMPH treatment. These observations support the existence of an unanticipated intracellular target that mediates the effects of AMPH on RhoA and cAMP signaling and suggest new pathways to target to disrupt AMPH action. PMID:26553986

  16. Rho GTPase protein Cdc42 is critical for postnatal cartilage development

    Energy Technology Data Exchange (ETDEWEB)

    Nagahama, Ryo [Department of Biochemistry, School of Dentistry, Showa University, Tokyo (Japan); Department of Orthodontics, School of Dentistry, Showa University, Tokyo (Japan); Yamada, Atsushi, E-mail: yamadaa@dent.showa-u.ac.jp [Department of Biochemistry, School of Dentistry, Showa University, Tokyo (Japan); Tanaka, Junichi [Department of Oral Diagnostic Sciences, School of Dentistry, Showa University, Tokyo (Japan); Aizawa, Ryo [Department of Periodontology, School of Dentistry, Showa University, Tokyo (Japan); Suzuki, Dai [Department of Biochemistry, School of Dentistry, Showa University, Tokyo (Japan); Kassai, Hidetoshi [Laboratory of Animal Resources, Center for Disease Biology and Integrative Medicine, Faculty of Medicine, The University of Tokyo, Tokyo (Japan); Yamamoto, Matsuo [Department of Periodontology, School of Dentistry, Showa University, Tokyo (Japan); Mishima, Kenji [Department of Oral Diagnostic Sciences, School of Dentistry, Showa University, Tokyo (Japan); Aiba, Atsu [Laboratory of Animal Resources, Center for Disease Biology and Integrative Medicine, Faculty of Medicine, The University of Tokyo, Tokyo (Japan); Maki, Koutaro [Department of Orthodontics, School of Dentistry, Showa University, Tokyo (Japan); Kamijo, Ryutaro [Department of Biochemistry, School of Dentistry, Showa University, Tokyo (Japan)

    2016-02-19

    Cdc42, a small Rho GTPase family member, has been shown to regulate multiple cellular functions in vitro, including actin cytoskeletal reorganization, cell migration, proliferation, and gene expression. However, its tissue-specific roles in vivo remain largely unknown, especially in postnatal cartilage development, as cartilage-specific Cdc42 inactivated mice die within a few days after birth. In this study, we investigated the physiological functions of Cdc42 during cartilage development after birth using tamoxifen-induced cartilage-specific inactivated Cdc42 conditional knockout (Cdc42 {sup fl/fl}; Col2-CreERT) mice, which were generated by crossing Cdc42 flox mice (Cdc42 {sup fl/fl}) with tamoxifen-induced type II collagen (Col2) Cre transgenic mice using a Cre/loxP system. The gross morphology of the Cdc42 cKO mice was shorter limbs and body, as well as reduced body weight as compared with the controls. In addition, severe defects were found in growth plate chondrocytes of the long bones, characterized by a shorter proliferating zone (PZ), wider hypertrophic zone (HZ), and loss of columnar organization of proliferating chondrocytes, resulting in delayed endochondral bone formation associated with abnormal bone growth. Our findings demonstrate the importance of Cdc42 for cartilage development during both embryonic and postnatal stages. - Highlights: • Tamoxifen-induced cartilage specific inactivated Cdc42 mutant mice were generated. • Cdc42 mutant mice were shorter limbs and body. • Severe defects were found in growth plate chondrocytes.

  17. The desorption of caesium from Peach Bottom HTGR steam generator materials

    International Nuclear Information System (INIS)

    Clark, M.J.

    1979-03-01

    The work at Harwell on the Peach Bottom End-of-Life Program in co-operation with the General Atomic Company (U.S.A.) is described. Materials taken from the Economiser, Evaporator and Superheater Sections of the Peach Bottom Unit No. 1. High Temperature Gas Cooled Reactor (HTGR) Heat Exchanger were placed in a reducing atmosphere comparable to the composition of an HTGR helium coolant gas, and the desorption of caesium isotopes measured under known conditions of flow, temperature and oxygen pressure. (author)

  18. Neuronal Rho GTPase Rac1 elimination confers neuroprotection in a mouse model of permanent ischemic stroke

    DEFF Research Database (Denmark)

    Karabiyik, Cansu; Fernandes, Rui; Figueiredo, Francisco Rosário

    2018-01-01

    The Rho GTPase Rac1 is a multifunctional protein involved in distinct pathways ranging from development to pathology. The aim of the present study was to unravel the contribution of neuronal Rac1 in regulating the response to brain injury induced by permanent focal cerebral ischemia (pMCAO). Our ...

  19. Neuronal Rho GTPase Rac1 elimination confers neuroprotection in a mice model of permanent ischemic stroke

    DEFF Research Database (Denmark)

    Karabiyik, Cansu; Fernandes, Rui; Figueiredo, Francisci Rosário

    2017-01-01

    The Rho GTPase Rac1 is a multifunctional protein involved in distinct pathways ranging from development to pathology. The aim of the present study was to unravel the contribution of neuronal Rac1 in regulating the response to brain injury induced by permanent focal cerebral ischemia (pMCAO). Our ...

  20. Interaction between the p21ras GTPase activating protein and the insulin receptor

    NARCIS (Netherlands)

    Pronk, G.J.; Medema, R.H.; Burgering, B.M.T.; Clark, R.; McCormick, F.; Bos, J.L.

    1992-01-01

    We investigated the involvement of the p21ras-GTPase activating protein (GAP) in insulin-induced signal transduction. In cells overexpressing the insulin receptor, we did not observe association between GAP and the insulin receptor after insulin treatment nor the phosphorylation of GAP on tyrosine

  1. The immunity-related GTPase Irga6 dimerizes in a parallel head-to-head fashion.

    Science.gov (United States)

    Schulte, Kathrin; Pawlowski, Nikolaus; Faelber, Katja; Fröhlich, Chris; Howard, Jonathan; Daumke, Oliver

    2016-03-02

    The immunity-related GTPases (IRGs) constitute a powerful cell-autonomous resistance system against several intracellular pathogens. Irga6 is a dynamin-like protein that oligomerizes at the parasitophorous vacuolar membrane (PVM) of Toxoplasma gondii leading to its vesiculation. Based on a previous biochemical analysis, it has been proposed that the GTPase domains of Irga6 dimerize in an antiparallel fashion during oligomerization. We determined the crystal structure of an oligomerization-impaired Irga6 mutant bound to a non-hydrolyzable GTP analog. Contrary to the previous model, the structure shows that the GTPase domains dimerize in a parallel fashion. The nucleotides in the center of the interface participate in dimerization by forming symmetric contacts with each other and with the switch I region of the opposing Irga6 molecule. The latter contact appears to activate GTP hydrolysis by stabilizing the position of the catalytic glutamate 106 in switch I close to the active site. Further dimerization contacts involve switch II, the G4 helix and the trans stabilizing loop. The Irga6 structure features a parallel GTPase domain dimer, which appears to be a unifying feature of all dynamin and septin superfamily members. This study contributes important insights into the assembly and catalytic mechanisms of IRG proteins as prerequisite to understand their anti-microbial action.

  2. Occupational Allergy to Peach (Prunus persica) Tree Pollen and Potential Cross-Reactivity between Rosaceae Family Pollens.

    Science.gov (United States)

    Jiang, Nannan; Yin, Jia; Mak, Philip; Wen, Liping

    2015-10-01

    Orchard workers in north China are highly exposed to orchard pollens, especially peach and other Rosaceae family pollens during pollination season. The aim of this study was to investigate whether occupational allergy to peach tree pollen as a member of Rosaceae family is IgE-mediated and to evaluate the cross-reactivity among Rosaceae family pollens. Allergen skin test and conjunctival challenge test were performed; enzyme linked immune-sorbent assay (ELISA), inhibiting ELISA, western immunoblotting and inhibiting western immunoblotting were done with Rosaceae family orchard pollens, including peach, apricot, cherry, apple and pear tree pollens. Mass spectrometry was also performed to probe the main allergen component and cross-reactive protein. Sensitizations to peach pollen were found in both skin test and conjunctival challenge in the patients. Serum specific IgE to three pollens (peach, apricot and cherry) were detected through ELISA. When peach pollen used as solid phase, ELISA inhibition revealed other four kinds of pollens capable of inducing partial to strong inhibitions (45% to 87%), with the strongest inhibition belonging to apricot pollen (87%). Western blotting showed predominant IgE binding to a 20 KD protein among these pollens, which appeared to be a cross-reactive allergen component through western blotting inhibition. It was recognized as a protein homologous to glutathione s-transferase 16 from Arabidopsis thaliana. Peach and other Rosaceae family tree pollen may serve as a potential cause of IgE mediated occupational respiratory disease in orchard workers in north China.

  3. Airborne Thermal Imagery to Detect the Seasonal Evolution of Crop Water Status in Peach, Nectarine and Saturn Peach Orchards

    Directory of Open Access Journals (Sweden)

    Joaquim Bellvert

    2016-01-01

    Full Text Available In the current scenario of worldwide limited water supplies, conserving water is a major concern in agricultural areas. Characterizing within-orchard spatial heterogeneity in water requirements would assist in improving irrigation water use efficiency and conserve water. The crop water stress index (CWSI has been successfully used as a crop water status indicator in several fruit tree species. In this study, the CWSI was developed in three Prunus persica L. cultivars at different phenological stages of the 2012 to 2014 growing seasons, using canopy temperature measurements of well-watered trees. The CWSI was then remotely estimated using high-resolution thermal imagery acquired from an airborne platform and related to leaf water potential (ѰL throughout the season. The feasibility of mapping within-orchard spatial variability of ѰL from thermal imagery was also explored. Results indicated that CWSI can be calculated using a common non-water-stressed baseline (NWSB, upper and lower limits for the entire growing season and for the three studied cultivars. Nevertheless, a phenological effect was detected in the CWSI vs. ѰL relationships. For a specific given CWSI value, ѰL was more negative as the crop developed. This different seasonal response followed the same trend for the three studied cultivars. The approach presented in this study demonstrated that CWSI is a feasible method to assess the spatial variability of tree water status in heterogeneous orchards, and to derive ѰL maps throughout a complete growing season. A sensitivity analysis of varying pixel size showed that a pixel size of 0.8 m or less was needed for precise ѰL mapping of peach and nectarine orchards with a tree crown area between 3.0 to 5.0 m2.

  4. Visualization of the Activity of Rac1 Small GTPase in a Cell

    International Nuclear Information System (INIS)

    Higashi, Morihiro; Yu, Jianyong; Tsuchiya, Hiroshi; Saito, Teruyoshi; Oyama, Toshinao; Kawana, Hidetada; Kitagawa, Motoo; Tamaru, Jun-ichi; Harigaya, Kenichi

    2010-01-01

    Rho family G proteins including Rac regulate a variety of cellular functions, such as morphology, motility, and gene expression. Here we developed a fluorescence resonance energy transfer-based analysis in which we could monitor the activity of Rac1. To detect fluorescence resonance energy transfer, yellow fluorescent protein fused Rac1 and cyan fluorescent protein fused Cdc42-Rac1-interaction-binding domain of Pak1 protein were used as intermolecular probes of FRET. The fluorophores were separated with linear unmixing method. The fluorescence resonance energy transfer efficiency was measured by acceptor photobleaching assisted assay. With these methods, the Rac1 activity was visualized in a cell. The present findings indicate that this approach is sensitive enough to achieve results similar to those from ratiometric fluorescence resonance energy transfer analysis

  5. The small GTPase, Rap1, mediates CD31-induced integrin adhesion

    NARCIS (Netherlands)

    Reedquist, K. A.; Ross, E.; Koop, E. A.; Wolthuis, R. M.; Zwartkruis, F. J.; van Kooyk, Y.; Salmon, M.; Buckley, C. D.; Bos, J. L.

    2000-01-01

    Integrin-mediated leukocyte adhesion is a critical aspect of leukocyte function that is tightly regulated by diverse stimuli, including chemokines, antigen receptors, and adhesion receptors. How cellular signals from CD31 and other adhesion amplifiers are integrated with those from classical

  6. The transmembrane adaptor protein NTAL signals to mast cell cytoskeleton via the small GTPase Rho

    Czech Academy of Sciences Publication Activity Database

    Tůmová, Magda; Koffer, Anna; Šimíček, Michal; Dráberová, Lubica; Dráber, Petr

    2010-01-01

    Roč. 40, č. 11 (2010), s. 3235-3245 ISSN 0014-2980 R&D Projects: GA MŠk 1M0506; GA MŠk LC545; GA ČR(CZ) GD204/05/H023; GA ČR GA301/09/1826; GA ČR GAP302/10/1759; GA AV ČR KAN200520701 Institutional research plan: CEZ:AV0Z50520514 Keywords : cell activation * cytoskeleton * mast cells Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.942, year: 2010

  7. The contribution of stored malate and citrate to the substrate requirements of metabolism of ripening peach (Prunus persica L. Batsch) flesh is negligible. Implications for the occurrence of phosphoenolpyruvate carboxykinase and gluconeogenesis.

    Science.gov (United States)

    Famiani, Franco; Farinelli, Daniela; Moscatello, Stefano; Battistelli, Alberto; Leegood, Richard C; Walker, Robert P

    2016-04-01

    The first aim of this study was to determine the contribution of stored malate and citrate to the substrate requirements of metabolism in the ripening flesh of the peach (Prunus persica L. Batsch) cultivar Adriatica. In the flesh, stored malate accumulated before ripening could contribute little or nothing to the net substrate requirements of metabolism. This was because there was synthesis and not dissimilation of malate throughout ripening. Stored citrate could potentially contribute a very small amount (about 5.8%) of the substrate required by metabolism when the whole ripening period was considered, and a maximum of about 7.5% over the latter part of ripening. The second aim of this study was to investigate why phosphoenolpyruvate carboxykinase (PEPCK) an enzyme utilised in gluconeogenesis from malate and citrate is present in peach flesh. The occurrence and localisation of enzymes utilised in the metabolism of malate, citrate and amino acids were determined in peach flesh throughout its development. Phosphoenolpyruvate carboxylase (essential for the synthesis of malate and citrate) was present in the same cells and at the same time as PEPCK and NADP-malic enzyme (both utilised in the dissimilation of malate and citrate). A hypothesis is presented to explain the presence of these enzymes and to account for the likely occurrence of gluconeogenesis. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  8. GTPase activity plays a key role in the pathobiology of LRRK2.

    Directory of Open Access Journals (Sweden)

    Yulan Xiong

    2010-04-01

    Full Text Available Mutations in the leucine-rich repeat kinase 2 (LRRK2 gene are associated with late-onset, autosomal-dominant, familial Parkinson's disease (PD and also contribute to sporadic disease. The LRRK2 gene encodes a large protein with multiple domains, including functional Roc GTPase and protein kinase domains. Mutations in LRRK2 most likely cause disease through a toxic gain-of-function mechanism. The expression of human LRRK2 variants in cultured primary neurons induces toxicity that is dependent on intact GTP binding or kinase activities. However, the mechanism(s underlying LRRK2-induced neuronal toxicity is poorly understood, and the contribution of GTPase and/or kinase activity to LRRK2 pathobiology is not well defined. To explore the pathobiology of LRRK2, we have developed a model of LRRK2 cytotoxicity in the baker's yeast Saccharomyces cerevisiae. Protein domain analysis in this model reveals that expression of GTPase domain-containing fragments of human LRRK2 are toxic. LRRK2 toxicity in yeast can be modulated by altering GTPase activity and is closely associated with defects in endocytic vesicular trafficking and autophagy. These truncated LRRK2 variants induce similar toxicity in both yeast and primary neuronal models and cause similar vesicular defects in yeast as full-length LRRK2 causes in primary neurons. The toxicity induced by truncated LRRK2 variants in yeast acts through a mechanism distinct from toxicity induced by human alpha-synuclein. A genome-wide genetic screen identified modifiers of LRRK2-induced toxicity in yeast including components of vesicular trafficking pathways, which can also modulate the trafficking defects caused by expression of truncated LRRK2 variants. Our results provide insight into the basic pathobiology of LRRK2 and suggest that the GTPase domain may contribute to the toxicity of LRRK2. These findings may guide future therapeutic strategies aimed at attenuating LRRK2-mediated neurodegeneration.

  9. Analysis of core damage frequency from internal events: Peach Bottom, Unit 2

    International Nuclear Information System (INIS)

    Kolaczkowski, A.M.; Lambright, J.A.; Ferrell, W.L.; Cathey, N.G.; Najafi, B.; Harper, F.T.

    1986-10-01

    This document contains the internal event initiated accident sequence analyses for Peach Bottom, Unit 2; one of the reference plants being examined as part of the NUREG-1150 effort by the Nuclear Regulatory Commission. NUREG-1150 will document the risk of a selected group of nuclear power plants. As part of that work, this report contains the overall core damage frequency estimate for Peach Bottom, Unit 2, and the accompanying plant damage state frequencies. Sensitivity and uncertainty analyses provided additional insights regarding the dominant contributors to the Peach Bottom core damage frequency estimate. The mean core damage frequency at Peach Bottom was calculated to be 8.2E-6. Station blackout type accidents (loss of all ac power) were found to dominate the overall results. Anticipated Transient Without Scram accidents were also found to be non-negligible contributors. The numerical results are largely driven by common mode failure probability estimates and to some extent, human error. Because of significant data and analysis uncertainties in these two areas (important, for instance, to the most dominant scenario in this study), it is recommended that the results of the uncertainty and sensitivity analyses be considered before any actions are taken based on this analysis

  10. Somatic Embryogenesis in Peach-Palm (Bactris gasipaes) Using Different Explant Sources.

    Science.gov (United States)

    Steinmacher, Douglas A; Heringer, Angelo Schuabb; Jiménez, Víctor M; Quoirin, Marguerite G G; Guerra, Miguel P

    2016-01-01

    Peach palm (Bactris gasipaes Kunth) is a member of the family Arecaceae and is a multipurpose but underutilized species. Nowadays, fruit production for subsistence and local markets, and heart-of-palm production for local, national, and international markets are the most important uses of this plant. Conventional breeding programs in peach palm are long-term efforts due to the prolonged generation time, large plant size, difficulties with controlled pollination and other factors. Although it is a caespitose palm, its propagation is currently based on seeds, as off-shoots are difficult to root. Hence, tissue culture techniques are considered to be the most likely strategy for efficient clonal plantlet regeneration of this species. Among various techniques, somatic embryogenesis offers the advantages of potential automated large-scale production and putative genetic stability of the regenerated plantlets. The induction of somatic embryogenesis in peach palm can be achieved by using different explant sources including zygotic embryos, immature inflorescences and thin cell layers from the young leaves and shoot meristems. The choice of a particular explant depends on whether clonal propagation is desired or not, as well as on the plant conditions and availability of explants. Protocols to induce and express somatic embryogenesis from different peach palm explants, up to acclimatization of plantlets, are described in this chapter.

  11. Characterising root density of peach trees in a semi-arid Chernozem to increase plant density

    Science.gov (United States)

    Paltineanu, Cristian; Septar, Leinar; Gavat, Corina; Chitu, Emil; Oprita, Alexandru; Moale, Cristina; Calciu, Irina; Vizitiu, Olga; Lamureanu, Gheorghe

    2016-01-01

    The available information on root system in fully mature peach orchards in semi-arid regions is insufficient. This paper presents a study on the root system density in an irrigated peach orchard from Dobrogea, Romania, using the trench technique. The old orchard has clean cultivation in inter-row and in-row. The objectives of the study were to: test the hypothesis that the roots of fully mature peach trees occupy the whole soil volume; find out if root repulsive effect of adjacent plants occurred for the rootstocks and soil conditions; find relationships between root system and soil properties and analyse soil state trend. Some soil physical properties were significantly deteriorated in inter-row versus in-row, mainly due to soil compaction induced by technological traffic. Density of total roots was higher in-row than inter-row, but the differences were not significant. Root density decreased more intensely with soil depth than with distance from tree trunks. Root density correlated with some soil properties. No repulsive effect of the roots of adjacent peach trees was noted. The decrease of root density with distance from trunk can be used in optimising tree arrangement. The conclusions could also be used in countries with similar growth conditions.

  12. Phytoplasmas in apricot, peach and sour cherry orchards in East Bohemia, Czech Republic

    Czech Academy of Sciences Publication Activity Database

    Ludvíková, H.; Fránová, Jana; Suchá, J.

    2011-01-01

    Roč. 64, Suppl. (2011), s. 67-68 ISSN 1721-8861 R&D Projects: GA MŠk OC09021 Institutional research plan: CEZ:AV0Z50510513 Keywords : Candidatus Phytoplasma prunorum * PCR/RFLP * apricot, peach and sour cherry orchards Subject RIV: EE - Microbiology, Virology Impact factor: 0.592, year: 2011

  13. Operating history report for the Peach Bottom HTGR. Volume I. Reactor operating history

    International Nuclear Information System (INIS)

    Scheffel, W.J.; Baldwin, N.L.; Tomlin, R.W.

    1976-01-01

    The operating history for the Peach Bottom-1 Reactor is presented for the years 1966 through 1975. Information concerning general chemistry data, general physics data, location of sensing elements in the primary helium circuit, and postirradiation examination and testing of reactor components is presented

  14. Fission product behavior in the Peach Bottom and Fort St. Vrain HTGRs

    International Nuclear Information System (INIS)

    Hanson, D.L.; Baldwin, N.L.; Strong, D.E.

    1980-11-01

    Actual operating data from Peach Bottom and Fort St. Vrain were compared with code predictions to assess the validity of the methods used to predict the behavior of fission products in the primary coolant circuit. For both reactors the measured circuit activities were significantly below design values, and the observations generally verify the codes used for large HTGR design

  15. Carbohydrate metabolism of vegetative and reproductive sinks in the late-maturing peach cultivar 'Encore'

    Science.gov (United States)

    Riccardo Lo Bianco; Mark Rieger; Shi-Jean S. Sung

    1999-01-01

    Activities of NAD+-dependent sorbitol dehydrogenase (SDH), sorbitol oxidase (SOX), sucrose synthase (SS), acid invertase (AI), and neutral invertase (NI) in ?Encore? peach (Prunus persica L.) fruits and developing shoot tips were assayed during the growing season to determine whether carbohydrate metabolizing enzymes could...

  16. Environmental effects on fruit ripening and average fruit weight for three peach cultivars

    Science.gov (United States)

    Three peach cultivars, ‘Crimson Lady’ (early), ‘Redhaven’ (mid-season) and ‘Cresthaven’ (late), were planted at twelve locations within the USA in 2009. All trees were grafted on ‘Lovell’ rootstock and came from the same nursery. Five trees of each cultivar were planted at a spacing of 6m by 5m at e...

  17. Response of Green Peach Aphids and Other Arthropods to Garlic Intercropped with Tobacco

    NARCIS (Netherlands)

    Lai, R.; You, M.; Lotz, L.A.P.; Vasseur, L.

    2011-01-01

    The green peach aphid, Myzus persicae (Sulzer), is an insect pest that causes extensive damage to tobacco (Nicotiana tabacum L.) in China. Field trials were conducted in 2008 and 2009 at Longyan in the Fujian Province (China) to evaluate the effects of garlic (Allium sativum L.) as a deterrent to

  18. DNA-based identification of Armillaria isolates from peach orchards in Mexico state

    Science.gov (United States)

    Ruben Damian Elias Roman; Ned B. Klopfenstein; Dionicio Alvarado Rosales; Mee-Sook Kim; Anna E. Case; Sara M. Ashiglar; John W. Hanna; Amy L. Ross-Davis; Remigio A. Guzman Plazola

    2012-01-01

    A collaborative project between the Programa de Fitopatología, Colegio de Postgraduados, Texcoco, Estado de Mexico and the USDA Forest Service - RMRS, Moscow Forest Pathology Laboratory has begun this year (2011) to assess which species of Armillaria are causing widespread and severe damage to the peach orchards from México state, Mexico. We are employing a DNA-based...

  19. A Non-Targeted Approach Unravels the Volatile Network in Peach Fruit

    Science.gov (United States)

    Sánchez, Gerardo; Besada, Cristina; Badenes, María Luisa; Monforte, Antonio José; Granell, Antonio

    2012-01-01

    Volatile compounds represent an important part of the plant metabolome and are of particular agronomic and biological interest due to their contribution to fruit aroma and flavor and therefore to fruit quality. By using a non-targeted approach based on HS-SPME-GC-MS, the volatile-compound complement of peach fruit was described. A total of 110 volatile compounds (including alcohols, ketones, aldehydes, esters, lactones, carboxylic acids, phenolics and terpenoids) were identified and quantified in peach fruit samples from different genetic backgrounds, locations, maturity stages and physiological responses. By using a combination of hierarchical cluster analysis and metabolomic correlation network analysis we found that previously known peach fruit volatiles are clustered according to their chemical nature or known biosynthetic pathways. Moreover, novel volatiles that had not yet been described in peach were identified and assigned to co-regulated groups. In addition, our analyses showed that most of the co-regulated groups showed good intergroup correlations that are therefore consistent with the existence of a higher level of regulation orchestrating volatile production under different conditions and/or developmental stages. In addition, this volatile network of interactions provides the ground information for future biochemical studies as well as a useful route map for breeding or biotechnological purposes. PMID:22761719

  20. RETRAN-3D MOD003 Peach Bottom Turbine Trip 2 Multidimensional Kinetics Analysis Models and Results

    International Nuclear Information System (INIS)

    Mori, Michitsugu; Ogura, Katsunori; Gose, Garry C.; Wu, J.-Y.

    2003-01-01

    An analysis of the Peach Bottom Unit 2 Turbine Trip Test 2 (PB2/TT2) has been performed using RETRAN-3D MOD003. The purpose of the analysis was to investigate the PB2/TT2 overpressurization transient using the RETRAN-3D multidimensional kinetics model

  1. An integrated approach for increasing breeding efficiency in apple and peach in Europe

    NARCIS (Netherlands)

    Laurens, Francois; Aranzana, Maria José; Arus, Pere; Bassi, Daniele; Bink, Marco; Bonany, Joan; Caprera, Andrea; Corelli-Grappadelli, Luca; Costes, Evelyne; Durel, Charles Eric; Mauroux, Jehan Baptiste; Muranty, Hélène; Nazzicari, Nelson; Pascal, Thierry; Patocchi, Andrea; Peil, Andreas; Quilot-Turion, Bénédicte; Rossini, Laura; Stella, Alessandra; Troggio, Michela; Velasco, Riccardo; De Weg, Van Eric

    2018-01-01

    Despite the availability of whole genome sequences of apple and peach, there has been a considerable gap between genomics and breeding. To bridge the gap, the European Union funded the FruitBreedomics project (March 2011 to August 2015) involving 28 research institutes and private companies. Three

  2. Modified atmosphere packaging extending the storage life of 'douradão' peach

    Directory of Open Access Journals (Sweden)

    Ligia Regina Radomille de Santana

    2010-12-01

    Full Text Available 'Douradão' peach is a perishable product and when cold stored is subject to chilling injury. The objective of the experiment was to evaluate the effect of modified atmosphere packaging (MAP and cold storage on quality and storage life of these peaches. Fruits were packed in polypropylene (PP trays and placed inside low density polyethylene (LDPE bags (30, 50, 60, 75 μm thickness with active modified atmosphere (10 kPa CO2 + 1.5kPa O2, balance N2. The control was made with peaches held in nonwrapped PP trays. Fruits were kept at 1 ± 1 °C and 90 ± 5% relative humidity (RH for 28 days and CO2 and O2 within packages was monitored every two days. After 14, 21 and 28 days, samples were withdrawn from MAP and kept in air at 25 ± 1 °C and 90 ± 5% RH for ripening. On the day of removal from the cold storage and after 4 days, peaches were evaluated for weight loss, decay incidence, flesh firmness, woolliness incidence, soluble solids content (SSC, titratable acidity (TA and juice content. The results showed that MAP had influence on reducing weight loss and prevented postharvest decay. MAP of 1-2 kPa O2 and 3-6 kPa CO2 at 1 °C (from 50 and 60 μm LDPE films were effective for keeping good quality of 'Douradão' peaches during 28 days of storage, the ripe fruits showed reduced incidence of woolliness, adequate juiciness and flesh firmness. Packages of 30 and 75 μm LDPE films were ineffective for reducing woolliness during cold storage. MAP fruits showed lower SSC and no relevant effect on TA. Control fruits did not present marketable conditions after 14 days of cold storage.

  3. New insights into the properties of pubescent surfaces: peach fruit as a model.

    Science.gov (United States)

    Fernández, Victoria; Khayet, Mohamed; Montero-Prado, Pablo; Heredia-Guerrero, José Alejandro; Liakopoulos, Georgios; Karabourniotis, George; Del Río, Víctor; Domínguez, Eva; Tacchini, Ignacio; Nerín, Cristina; Val, Jesús; Heredia, Antonio

    2011-08-01

    The surface of peach (Prunus persica 'Calrico') is covered by a dense indumentum, which may serve various protective purposes. With the aim of relating structure to function, the chemical composition, morphology, and hydrophobicity of the peach skin was assessed as a model for a pubescent plant surface. Distinct physicochemical features were observed for trichomes versus isolated cuticles. Peach cuticles were composed of 53% cutan, 27% waxes, 23% cutin, and 1% hydroxycinnamic acid derivatives (mainly ferulic and p-coumaric acids). Trichomes were covered by a thin cuticular layer containing 15% waxes and 19% cutin and were filled by polysaccharide material (63%) containing hydroxycinnamic acid derivatives and flavonoids. The surface free energy, polarity, and work of adhesion of intact and shaved peach surfaces were calculated from contact angle measurements of water, glycerol, and diiodomethane. The removal of the trichomes from the surface increased polarity from 3.8% (intact surface) to 23.6% and decreased the total surface free energy chiefly due to a decrease on its nonpolar component. The extraction of waxes and the removal of trichomes led to higher fruit dehydration rates. However, trichomes were found to have a higher water sorption capacity as compared with isolated cuticles. The results show that the peach surface is composed of two different materials that establish a polarity gradient: the trichome network, which has a higher surface free energy and a higher dispersive component, and the cuticle underneath, which has a lower surface free energy and higher surface polarity. The significance of the data concerning water-plant surface interactions is discussed within a physiological context.

  4. Comparative Transcriptome and Microscopy Analyses Provide Insights into Flat Shape Formation in Peach (Prunus persica

    Directory of Open Access Journals (Sweden)

    Jian Guo

    2018-01-01

    Full Text Available Fruit shape is an important external characteristic that consumers use to select preferred fruit cultivars. In peach, the flat fruit cultivars have become more and more popular worldwide. Genetic markers closely linking to the flat fruit trait have been identified and are useful for marker-assisted breeding. However, the cellular and genetic mechanisms underpinning flat fruit formation are still poorly understood. In this study, we have revealed the differences in fruit cell number, cell size, and in gene expression pattern between the traditional round fruit and modern flat fruit cultivars. Flat peach cultivars possessed significantly lower number of cells in the vertical axis because cell division in the vertical direction stopped early in the flat fruit cultivars at 15 DAFB (day after full bloom than in round fruit cultivars at 35 DAFB. This resulted in the reduction in vertical development in the flat fruit. Significant linear relationship was observed between fruit vertical diameter and cell number in vertical axis for the four examined peach cultivars (R2 = 0.9964 at maturation stage, and was also observed between fruit vertical diameter and fruit weight (R2 = 0.9605, which indicated that cell number in vertical direction contributed to the flat shape formation. Furthermore, in RNA-seq analysis, 4165 differentially expressed genes (DEGs were detected by comparing RNA-seq data between flat and round peach cultivars at different fruit development stages. In contrast to previous studies, we discovered 28 candidate genes potentially responsible for the flat shape formation, including 19 located in the mapping site and 9 downstream genes. Our study indicates that flat and round fruit shape in peach is primarily determined by the regulation of cell production in the vertical direction during early fruit development.

  5. Multivalent adhesion molecule 7 clusters act as signaling platform for host cellular GTPase activation and facilitate epithelial barrier dysfunction.

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    Jenson Lim

    2014-09-01

    Full Text Available Vibrio parahaemolyticus is an emerging bacterial pathogen which colonizes the gastrointestinal tract and can cause severe enteritis and bacteraemia. During infection, V. parahaemolyticus primarily attaches to the small intestine, where it causes extensive tissue damage and compromises epithelial barrier integrity. We have previously described that Multivalent Adhesion Molecule (MAM 7 contributes to initial attachment of V. parahaemolyticus to epithelial cells. Here we show that the bacterial adhesin, through multivalent interactions between surface-induced adhesin clusters and phosphatidic acid lipids in the host cell membrane, induces activation of the small GTPase RhoA and actin rearrangements in host cells. In infection studies with V. parahaemolyticus we further demonstrate that adhesin-triggered activation of the ROCK/LIMK signaling axis is sufficient to redistribute tight junction proteins, leading to a loss of epithelial barrier function. Taken together, these findings show an unprecedented mechanism by which an adhesin acts as assembly platform for a host cellular signaling pathway, which ultimately facilitates breaching of the epithelial barrier by a bacterial pathogen.

  6. Novel Activities of Select NSAID R-Enantiomers against Rac1 and Cdc42 GTPases.

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    Tudor I Oprea

    Full Text Available Rho family GTPases (including Rac, Rho and Cdc42 collectively control cell proliferation, adhesion and migration and are of interest as functional therapeutic targets in numerous epithelial cancers. Based on high throughput screening of the Prestwick Chemical Library® and cheminformatics we identified the R-enantiomers of two approved drugs (naproxen and ketorolac as inhibitors of Rac1 and Cdc42. The corresponding S-enantiomers are considered the active component in racemic drug formulations, acting as non-steroidal anti-inflammatory drugs (NSAIDs with selective activity against cyclooxygenases. Here, we show that the S-enantiomers of naproxen and ketorolac are inactive against the GTPases. Additionally, more than twenty other NSAIDs lacked inhibitory action against the GTPases, establishing the selectivity of the two identified NSAIDs. R-naproxen was first identified as a lead compound and tested in parallel with its S-enantiomer and the non-chiral 6-methoxy-naphthalene acetic acid (active metabolite of nabumetone, another NSAID as a structural series. Cheminformatics-based substructure analyses-using the rotationally constrained carboxylate in R-naproxen-led to identification of racemic [R/S] ketorolac as a suitable FDA-approved candidate. Cell based measurement of GTPase activity (in animal and human cell lines demonstrated that the R-enantiomers specifically inhibit epidermal growth factor stimulated Rac1 and Cdc42 activation. The GTPase inhibitory effects of the R-enantiomers in cells largely mimic those of established Rac1 (NSC23766 and Cdc42 (CID2950007/ML141 specific inhibitors. Docking predicts that rotational constraints position the carboxylate moieties of the R-enantiomers to preferentially coordinate the magnesium ion, thereby destabilizing nucleotide binding to Rac1 and Cdc42. The S-enantiomers can be docked but are less favorably positioned in proximity to the magnesium. R-naproxen and R-ketorolac have potential for rapid

  7. Pre-symptomatic transcriptome changes during cold storage of chilling sensitive and resistant peach cultivars to elucidate chilling injury mechanisms

    OpenAIRE

    Puig, Clara Pons; Dagar, Anurag; Marti Ibanez, Cristina; Singh, Vikram; Crisosto, Carlos H; Friedman, Haya; Lurie, Susan; Granell, Antonio

    2015-01-01

    Background: Cold storage induces chilling injury (CI) disorders in peach fruit (woolliness/mealiness, flesh browning and reddening/bleeding) manifested when ripened at shelf life. To gain insight into the mechanisms underlying CI, we analyzed the transcriptome of 'Oded' (high tolerant) and 'Hermoza' (relatively tolerant to woolliness, but sensitive to browning and bleeding) peach cultivars at pre-symptomatic stages. The expression profiles were compared and validated with two previously analy...

  8. Characterization and Functional Analysis of the Calmodulin-Binding Domain of Rac1 GTPase

    Science.gov (United States)

    Xu, Bing; Chelikani, Prashen; Bhullar, Rajinder P.

    2012-01-01

    Rac1, a member of the Rho family of small GTPases, has been shown to promote formation of lamellipodia at the leading edge of motile cells and affect cell migration. We previously demonstrated that calmodulin can bind to a region in the C-terminal of Rac1 and that this interaction is important in the activation of platelet Rac1. Now, we have analyzed amino acid residue(s) in the Rac1-calmodulin binding domain that are essential for the interaction and assessed their functional contribution in Rac1 activation. The results demonstrated that region 151–164 in Rac1 is essential for calmodulin binding. Within the 151–164 region, positively-charged amino acids K153 and R163 were mutated to alanine to study impact on calmodulin binding. Mutant form of Rac1 (K153A) demonstrated significantly reduced binding to calmodulin while the double mutant K153A/R163A demonstrated complete lack of binding to calmodulin. Thrombin or EGF resulted in activation of Rac1 in CHRF-288-11 or HeLa cells respectively and W7 inhibited this activation. Immunoprecipitation studies demonstrated that higher amount of CaM was associated with Rac1 during EGF dependent activation. In cells expressing mutant forms of Rac1 (K153A or K153A/R163A), activation induced by EGF was significantly decreased in comparison to wild type or the R163A forms of Rac1. The lack of Rac1 activation in mutant forms was not due to an inability of GDP-GTP exchange or a change in subcelllular distribution. Moreover, Rac1 activation was decreased in cells where endogenous level of calmodulin was reduced using shRNA knockdown and increased in cells where calmodulin was overexpressed. Docking analysis and modeling demonstrated that K153 in Rac1 interacts with Q41 in calmodulin. These results suggest an important role for calmodulin in the activation of Rac1 and thus, in cytoskeleton reorganization and cell migration. PMID:22905193

  9. Characterization and functional analysis of the calmodulin-binding domain of Rac1 GTPase.

    Directory of Open Access Journals (Sweden)

    Bing Xu

    Full Text Available Rac1, a member of the Rho family of small GTPases, has been shown to promote formation of lamellipodia at the leading edge of motile cells and affect cell migration. We previously demonstrated that calmodulin can bind to a region in the C-terminal of Rac1 and that this interaction is important in the activation of platelet Rac1. Now, we have analyzed amino acid residue(s in the Rac1-calmodulin binding domain that are essential for the interaction and assessed their functional contribution in Rac1 activation. The results demonstrated that region 151-164 in Rac1 is essential for calmodulin binding. Within the 151-164 region, positively-charged amino acids K153 and R163 were mutated to alanine to study impact on calmodulin binding. Mutant form of Rac1 (K153A demonstrated significantly reduced binding to calmodulin while the double mutant K153A/R163A demonstrated complete lack of binding to calmodulin. Thrombin or EGF resulted in activation of Rac1 in CHRF-288-11 or HeLa cells respectively and W7 inhibited this activation. Immunoprecipitation studies demonstrated that higher amount of CaM was associated with Rac1 during EGF dependent activation. In cells expressing mutant forms of Rac1 (K153A or K153A/R163A, activation induced by EGF was significantly decreased in comparison to wild type or the R163A forms of Rac1. The lack of Rac1 activation in mutant forms was not due to an inability of GDP-GTP exchange or a change in subcelllular distribution. Moreover, Rac1 activation was decreased in cells where endogenous level of calmodulin was reduced using shRNA knockdown and increased in cells where calmodulin was overexpressed. Docking analysis and modeling demonstrated that K153 in Rac1 interacts with Q41 in calmodulin. These results suggest an important role for calmodulin in the activation of Rac1 and thus, in cytoskeleton reorganization and cell migration.

  10. WAVE regulatory complex activation by cooperating GTPases Arf and Rac1

    DEFF Research Database (Denmark)

    Koronakis, Vassilis; Hume, Peter J; Humphreys, Daniel

    2011-01-01

    The WAVE regulatory complex (WRC) is a critical element in the control of actin polymerization at the eukaryotic cell membrane, but how WRC is activated remains uncertain. While Rho GTPase Rac1 can bind and activate WRC in vitro, this interaction is of low affinity, suggesting other factors may...... be important. By reconstituting WAVE-dependent actin assembly on membrane-coated beads in mammalian cell extracts, we found that Rac1 was not sufficient to engender bead motility, and we uncovered a key requirement for Arf GTPases. In vitro, Rac1 and Arf1 were individually able to bind weakly to recombinant...... be central components in WAVE signalling, acting directly, alongside Rac1....

  11. Nanofibrillar scaffolds induce preferential activation of Rho GTPases in cerebral cortical astrocytes

    Science.gov (United States)

    Tiryaki, Volkan Mujdat; Ayres, Virginia M; Khan, Adeel A; Ahmed, Ijaz; Shreiber, David I; Meiners, Sally

    2012-01-01

    Cerebral cortical astrocyte responses to polyamide nanofibrillar scaffolds versus poly-L-lysine (PLL)-functionalized planar glass, unfunctionalized planar Aclar coverslips, and PLL-functionalized planar Aclar surfaces were investigated by atomic force microscopy and immunocytochemistry. The physical properties of the cell culture environments were evaluated using contact angle and surface roughness measurements and compared. Astrocyte morphological responses, including filopodia, lamellipodia, and stress fiber formation, and stellation were imaged using atomic force microscopy and phalloidin staining for F-actin. Activation of the corresponding Rho GTPase regulators was investigated using immunolabeling with Cdc42, Rac1, and RhoA. Astrocytes cultured on the nanofibrillar scaffolds showed a unique response that included stellation, cell–cell interactions by stellate processes, and evidence of depression of RhoA. The results support the hypothesis that the extracellular environment can trigger preferential activation of members of the Rho GTPase family, with demonstrable morphological consequences for cerebral cortical astrocytes. PMID:22915841

  12. Rab GTPases Regulate Endothelial Cell Protein C Receptor-Mediated Endocytosis and Trafficking of Factor VIIa

    Science.gov (United States)

    Nayak, Ramesh C.; Keshava, Shiva; Esmon, Charles T.; Pendurthi, Usha R.; Rao, L. Vijaya Mohan

    2013-01-01

    Recent studies have established that factor VIIa (FVIIa) binds to the endothelial cell protein C receptor (EPCR). FVIIa binding to EPCR may promote the endocytosis of this receptor/ligand complex. Rab GTPases are known to play a crucial role in the endocytic and exocytic pathways of receptors or receptor/ligand complexes. The present study was undertaken to investigate the role of Rab GTPases in the intracellular trafficking of EPCR and FVIIa. CHO-EPCR cells and human umbilical vein endothelial cells (HUVEC) were transduced with recombinant adenoviral vectors to express wild-type, constitutively active, or dominant negative mutant of various Rab GTPases. Cells were exposed to FVIIa conjugated with AF488 fluorescent probe (AF488-FVIIa), and intracellular trafficking of FVIIa, EPCR, and Rab proteins was evaluated by immunofluorescence confocal microscopy. In cells expressing wild-type or constitutively active Rab4A, internalized AF488-FVIIa accumulated in early/sorting endosomes and its entry into the recycling endosomal compartment (REC) was inhibited. Expression of constitutively active Rab5A induced large endosomal structures beneath the plasma membrane where EPCR and FVIIa accumulated. Dominant negative Rab5A inhibited the endocytosis of EPCR-FVIIa. Expression of constitutively active Rab11 resulted in retention of accumulated AF488-FVIIa in the REC, whereas expression of a dominant negative form of Rab11 led to accumulation of internalized FVIIa in the cytoplasm and prevented entry of internalized FVIIa into the REC. Expression of dominant negative Rab11 also inhibited the transport of FVIIa across the endothelium. Overall our data show that Rab GTPases regulate the internalization and intracellular trafficking of EPCR-FVIIa. PMID:23555015

  13. Insight into temperature dependence of GTPase activity in human guanylate binding protein-1.

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    Anjana Rani

    Full Text Available Interferon-γ induced human guanylate binding protein-1(hGBP1 belongs to a family of dynamin related large GTPases. Unlike all other GTPases, hGBP1 hydrolyzes GTP to a mixture of GDP and GMP with GMP being the major product at 37°C but GDP became significant when the hydrolysis reaction was carried out at 15°C. The hydrolysis reaction in hGBP1 is believed to involve with a number of catalytic steps. To investigate the effect of temperature in the product formation and on the different catalytic complexes of hGBP1, we carried out temperature dependent GTPase assays, mutational analysis, chemical and thermal denaturation studies. The Arrhenius plot for both GDP and GMP interestingly showed nonlinear behaviour, suggesting that the product formation from the GTP-bound enzyme complex is associated with at least more than one step. The negative activation energy for GDP formation and GTPase assay with external GDP together indicate that GDP formation occurs through the reversible dissociation of GDP-bound enzyme dimer to monomer, which further reversibly dissociates to give the product. Denaturation studies of different catalytic complexes show that unlike other complexes the free energy of GDP-bound hGBP1 decreases significantly at lower temperature. GDP formation is found to be dependent on the free energy of the GDP-bound enzyme complex. The decrease in the free energy of this complex at low temperature compared to at high is the reason for higher GDP formation at low temperature. Thermal denaturation studies also suggest that the difference in the free energy of the GTP-bound enzyme dimer compared to its monomer plays a crucial role in the product formation; higher stability favours GMP but lower favours GDP. Thus, this study provides the first thermodynamic insight into the effect of temperature in the product formation of hGBP1.

  14. Structure of the ribosomal interacting GTPase YjeQ from the enterobacterial species Salmonella typhimurium

    Energy Technology Data Exchange (ETDEWEB)

    Nichols, C. E. [Division of Structural Biology, The Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN (United Kingdom); Johnson, C.; Lamb, H. K. [Institute of Cell and Molecular Biosciences, Catherine Cookson Building, Medical School, Framlington Place, Newcastle University, Newcastle-upon-Tyne NE2 4HH (United Kingdom); Lockyer, M. [Arrow Therapeutics Ltd, Britannia House, Trinity Street, Borough, London SE1 1DA (United Kingdom); Charles, I. G. [The Wolfson Institute for Biomedical Research, The Cruciform Building, University College London, Gower Street, London WC1E 6BT (United Kingdom); Hawkins, A. R. [Institute of Cell and Molecular Biosciences, Catherine Cookson Building, Medical School, Framlington Place, Newcastle University, Newcastle-upon-Tyne NE2 4HH (United Kingdom); Stammers, D. K., E-mail: daves@strubi.ox.ac.uk [Division of Structural Biology, The Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN (United Kingdom)

    2007-11-01

    The X-ray crystal structure of the GTPase YjeQ from S. typhimurium is presented and compared with those of orthologues from T. maritima and B. subtilis. The YjeQ class of P-loop GTPases assist in ribosome biogenesis and also bind to the 30S subunit of mature ribosomes. YjeQ ribosomal binding is GTP-dependent and thought to specifically direct protein synthesis, although the nature of the upstream signal causing this event in vivo is as yet unknown. The attenuating effect of YjeQ mutants on bacterial growth in Escherichia coli makes it a potential target for novel antimicrobial agents. In order to further explore the structure and function of YjeQ, the isolation, crystallization and structure determination of YjeQ from the enterobacterial species Salmonella typhimurium (StYjeQ) is reported. Whilst the overall StYjeQ fold is similar to those of the previously reported Thematoga maritima and Bacillus subtilis orthologues, particularly the GTPase domain, there are larger differences in the three OB folds. Although the zinc-finger secondary structure is conserved, significant sequence differences alter the nature of the external surface in each case and may reflect varying signalling pathways. Therefore, it may be easier to develop YjeQ-specific inhibitors that target the N- and C-terminal regions, disrupting the metabolic connectivity rather than the GTPase activity. The availability of coordinates for StYjeQ will provide a significantly improved basis for threading Gram-negative orthologue sequences and in silico compound-screening studies, with the potential for the development of species-selective drugs.

  15. Structure of the ribosomal interacting GTPase YjeQ from the enterobacterial species Salmonella typhimurium

    International Nuclear Information System (INIS)

    Nichols, C. E.; Johnson, C.; Lamb, H. K.; Lockyer, M.; Charles, I. G.; Hawkins, A. R.; Stammers, D. K.

    2007-01-01

    The X-ray crystal structure of the GTPase YjeQ from S. typhimurium is presented and compared with those of orthologues from T. maritima and B. subtilis. The YjeQ class of P-loop GTPases assist in ribosome biogenesis and also bind to the 30S subunit of mature ribosomes. YjeQ ribosomal binding is GTP-dependent and thought to specifically direct protein synthesis, although the nature of the upstream signal causing this event in vivo is as yet unknown. The attenuating effect of YjeQ mutants on bacterial growth in Escherichia coli makes it a potential target for novel antimicrobial agents. In order to further explore the structure and function of YjeQ, the isolation, crystallization and structure determination of YjeQ from the enterobacterial species Salmonella typhimurium (StYjeQ) is reported. Whilst the overall StYjeQ fold is similar to those of the previously reported Thematoga maritima and Bacillus subtilis orthologues, particularly the GTPase domain, there are larger differences in the three OB folds. Although the zinc-finger secondary structure is conserved, significant sequence differences alter the nature of the external surface in each case and may reflect varying signalling pathways. Therefore, it may be easier to develop YjeQ-specific inhibitors that target the N- and C-terminal regions, disrupting the metabolic connectivity rather than the GTPase activity. The availability of coordinates for StYjeQ will provide a significantly improved basis for threading Gram-negative orthologue sequences and in silico compound-screening studies, with the potential for the development of species-selective drugs

  16. Slit-Robo GTPase-Activating Protein 2 as a metastasis suppressor in osteosarcoma

    OpenAIRE

    Marko, Tracy A.; Shamsan, Ghaidan A.; Edwards, Elizabeth N.; Hazelton, Paige E.; Rathe, Susan K.; Cornax, Ingrid; Overn, Paula R.; Varshney, Jyotika; Diessner, Brandon J.; Moriarity, Branden S.; O?Sullivan, M. Gerard; Odde, David J.; Largaespada, David A.

    2016-01-01

    Osteosarcoma is the most common primary bone tumor, with metastatic disease responsible for most treatment failure and patient death. A forward genetic screen utilizing Sleeping Beauty mutagenesis in mice previously identified potential genetic drivers of osteosarcoma metastasis, including Slit-Robo GTPase-Activating Protein 2 (Srgap2). This study evaluates the potential role of SRGAP2 in metastases-associated properties of osteosarcoma cell lines through Srgap2 knockout via the CRISPR/Cas9 n...

  17. Targeting GTPases in Parkinson’s disease: comparison to the historic path of kinase drug discovery and perspectives

    Directory of Open Access Journals (Sweden)

    LIN eHONG

    2014-06-01

    Full Text Available Neurological diseases have placed heavy social and financial burdens on modern society. As the life expectancy of humans is extended, neurological diseases, such as Parkinson’s disease, have become increasingly common among senior populations. Although the enigmas of Parkinson’s diseases await resolution, more vivid pictures on the cause, progression and control of the illness are emerging after years of research. On the molecular level, GTPases are implicated in the etiology of Parkinson’s disease and are rational pharmaceutical targets for their control. However, targeting individual GTPases, which belong to a superfamily of proteins containing multiple members with a conserved guanine nucleotide binding domain, has proven to be challenging. In contrast, pharmaceutical pursuit of inhibition of kinases, which constitute another superfamily of proteins with more than 500 members, has been fairly successful. We reviewed the breakthroughs in the history of kinase drug discovery to provide guidance for the GTPase field. We summarize recent progress made in the regulation of GTPase activity. We also present an efficient and cost effective approach to drug screening, which uses multiplex flow cytometry and mixture-based positional scanning libraries. These methods allow simultaneous measurements of both the activity and the selectivity of the screened library. Several GTPase activator clusters were identified which showed selectivity against different GTPase subfamilies. While the clusters need to be further deconvoluted to identify individual active compounds, the method described here and the structure information gathered create a foundation for further developments to build upon.

  18. A class of dynamin-like GTPases involved in the generation of the tubular ER network

    Science.gov (United States)

    Hu, Junjie; Shibata, Yoko; Zhu, Peng-Peng; Voss, Christiane; Rismanchi, Neggy; Prinz, William A.; Rapoport, Tom A.; Blackstone, Craig

    2009-01-01

    The endoplasmic reticulum (ER) consists of tubules that are shaped by the reticulons and DP1/Yop1p, but how the tubules form an interconnected network is unknown. Here, we show that mammalian atlastins, which are dynamin-like, integral membrane GTPases, interact with the tubule-shaping proteins. The atlastins localize to the tubular ER and are required for proper network formation in vivo and in vitro. Depletion of the atlastins or overexpression of dominant-negative forms inhibits tubule interconnections. The Sey1p GTPase in S. cerevisiae is likely a functional ortholog of the atlastins; it shares the same signature motifs and membrane topology and interacts genetically and physically with the tubule-shaping proteins. Cells simultaneously lacking Sey1p and a tubule-shaping protein have ER morphology defects. These results indicate that formation of the tubular ER network depends on conserved dynamin-like GTPases. Since atlastin-1 mutations cause a common form of hereditary spastic paraplegia, we suggest ER shaping defects as a novel neuropathogenic mechanism. PMID:19665976

  19. Influence of bacterial toxins on the GTPase activity of transducin from bovine retinal rod outer segments

    International Nuclear Information System (INIS)

    Rybin, V.O.; Gureeva, A.A.

    1986-01-01

    The action of cholera toxin, capable of ADP-ribosylation of the activator N/sub s/ protein, and pertussis toxin, capable of ADP-ribosylation of the inhibitor N/sub i/ protein of the adenylate cyclase complex, on transducin, the GTP-binding protein of the rod outer segments of the retina, was investigated. It was shown that under the action of pertussis and cholera toxins, the GTPase activity of transducin is inhibited. Pertussin toxin inhibits the GTPase of native retinal rod outer segments by 30-40%, while GTPase of homogeneous transducin produces a 70-80% inhibition. The action of toxins on transducin depends on the presence and nature of the guanylic nucleotide with which incubation is performed. On the basis of the data obtained it is suggested that pertussis toxin interacts with pretransducin and with the transducin-GDP complex, while cholera toxin ADP-ribosylates the transducin-GTP complex and does not act on transducin lacking GTP

  20. The GTPase Rab37 Participates in the Control of Insulin Exocytosis.

    Directory of Open Access Journals (Sweden)

    Sanda Ljubicic

    Full Text Available Rab37 belongs to a subclass of Rab GTPases regulating exocytosis, including also Rab3a and Rab27a. Proteomic studies indicate that Rab37 is associated with insulin-containing large dense core granules of pancreatic β-cells. In agreement with these observations, we detected Rab37 in extracts of β-cell lines and human pancreatic islets and confirmed by confocal microscopy the localization of the GTPase on insulin-containing secretory granules. We found that, as is the case for Rab3a and Rab27a, reduction of Rab37 levels by RNA interference leads to impairment in glucose-induced insulin secretion and to a decrease in the number of granules in close apposition to the plasma membrane. Pull-down experiments revealed that, despite similar functional effects, Rab37 does not interact with known Rab3a or Rab27a effectors and is likely to operate through a different mechanism. Exposure of insulin-secreting cells to proinflammatory cytokines, fatty acids or oxidized low-density lipoproteins, mimicking physiopathological conditions that favor the development of diabetes, resulted in a decrease in Rab37 expression. Our data identify Rab37 as an additional component of the machinery governing exocytosis of β-cells and suggest that impaired expression of this GTPase may contribute to defective insulin release in pre-diabetic and diabetic conditions.

  1. 9Å structure of the COPI coat reveals that the Arf1 GTPase occupies two contrasting molecular environments.

    Science.gov (United States)

    Dodonova, Svetlana O; Aderhold, Patrick; Kopp, Juergen; Ganeva, Iva; Röhling, Simone; Hagen, Wim J H; Sinning, Irmgard; Wieland, Felix; Briggs, John A G

    2017-06-16

    COPI coated vesicles mediate trafficking within the Golgi apparatus and between the Golgi and the endoplasmic reticulum. Assembly of a COPI coated vesicle is initiated by the small GTPase Arf1 that recruits the coatomer complex to the membrane, triggering polymerization and budding. The vesicle uncoats before fusion with a target membrane. Coat components are structurally conserved between COPI and clathrin/adaptor proteins. Using cryo-electron tomography and subtomogram averaging, we determined the structure of the COPI coat assembled on membranes in vitro at 9 Å resolution. We also obtained a 2.57 Å resolution crystal structure of βδ-COP. By combining these structures we built a molecular model of the coat. We additionally determined the coat structure in the presence of ArfGAP proteins that regulate coat dissociation. We found that Arf1 occupies contrasting molecular environments within the coat, leading us to hypothesize that some Arf1 molecules may regulate vesicle assembly while others regulate coat disassembly.

  2. Gene targeting implicates Cdc42 GTPase in GPVI and non-GPVI mediated platelet filopodia formation, secretion and aggregation.

    Directory of Open Access Journals (Sweden)

    Huzoor Akbar

    Full Text Available Cdc42 and Rac1, members of the Rho family of small GTPases, play critical roles in actin cytoskeleton regulation. We have shown previously that Rac1 is involved in regulation of platelet secretion and aggregation. However, the role of Cdc42 in platelet activation remains controversial. This study was undertaken to better understand the role of Cdc42 in platelet activation.We utilized the Mx-cre;Cdc42(lox/lox inducible mice with transient Cdc42 deletion to investigate the involvement of Cdc42 in platelet function. The Cdc42-deficient mice exhibited a significantly reduced platelet count than the matching Cdc42(+/+ mice. Platelets isolated from Cdc42(-/-, as compared to Cdc42(+/+, mice exhibited (a diminished phosphorylation of PAK1/2, an effector molecule of Cdc42, (b inhibition of filopodia formation on immobilized CRP or fibrinogen, (c inhibition of CRP- or thrombin-induced secretion of ATP and release of P-selectin, (d inhibition of CRP, collagen or thrombin induced platelet aggregation, and (e minimal phosphorylation of Akt upon stimulation with CRP or thrombin. The bleeding times were significantly prolonged in Cdc42(-/- mice compared with Cdc42(+/+ mice.Our data demonstrate that Cdc42 is required for platelet filopodia formation, secretion and aggregation and therefore plays a critical role in platelet mediated hemostasis and thrombosis.

  3. Rad GTPase is essential for the regulation of bone density and bone marrow adipose tissue in mice.

    Science.gov (United States)

    Withers, Catherine N; Brown, Drew M; Byiringiro, Innocent; Allen, Matthew R; Condon, Keith W; Satin, Jonathan; Andres, Douglas A

    2017-10-01

    The small GTP-binding protein Rad (RRAD, Ras associated with diabetes) is the founding member of the RGK (Rad, Rem, Rem2, and Gem/Kir) family that regulates cardiac voltage-gated Ca 2+ channel function. However, its cellular and physiological functions outside of the heart remain to be elucidated. Here we report that Rad GTPase function is required for normal bone homeostasis in mice, as Rad deletion results in significantly lower bone mass and higher bone marrow adipose tissue (BMAT) levels. Dynamic histomorphometry in vivo and primary calvarial osteoblast assays in vitro demonstrate that bone formation and osteoblast mineralization rates are depressed, while in vitro osteoclast differentiation is increased, in the absence of Rad. Microarray analysis revealed that canonical osteogenic gene expression (Runx2, osterix, etc.) is not altered in Rad -/- calvarial osteoblasts; instead robust up-regulation of matrix Gla protein (MGP, +11-fold), an inhibitor of extracellular matrix mineralization and a protein secreted during adipocyte differentiation, was observed. Strikingly, Rad deficiency also resulted in significantly higher marrow adipose tissue levels in vivo and promoted spontaneous in vitro adipogenesis of primary calvarial osteoblasts. Adipogenic differentiation of wildtype calvarial osteoblasts resulted in the loss of endogenous Rad protein, further supporting a role for Rad in the control of BMAT levels. These findings reveal a novel in vivo function for Rad and establish a role for Rad signaling in the complex physiological control of skeletal homeostasis and bone marrow adiposity. Copyright © 2017 Elsevier Inc. All rights reserved.

  4. C9orf72’s interaction with Rab GTPases - modulation of membrane traffic and autophagy

    Directory of Open Access Journals (Sweden)

    Bor Luen Tang

    2016-10-01

    Full Text Available Hexanucleotide repeat expansion in an intron of Chromosome 9 open reading frame 72 (C9orf72 is the most common genetic cause of Amyotrophic Lateral Sclerosis (ALS and Frontotemporal Dementia (FTD. While functional haploinsufficiency of C9orf72 resulting from the mutation may play a role in ALS/FTD, the actual cellular role of the protein has been unclear. Recent findings have now shown that C9orf72 physically and functionally interacts with multiple members of the Rab small GTPases family, consequently exerting important influences on cellular membrane traffic and the process of autophagy. Loss of C9orf72 impairs endocytosis in neuronal cell lines, and attenuated autophagosome formation. Interestingly, C9orf72 could influence autophagy both as part of a Guanine nucleotide exchange factor (GEF complex, or as a Rab effector that facilitates transport of the Unc-51-like Autophagy Activating Kinase 1 (Ulk1 autophagy initiation complex. The cellular function of C9orf72 is discussed in the light of these recent findings

  5. Hepatic trans-Golgi action coordinated by the GTPase ARFRP1 is crucial for lipoprotein lipidation and assembly[S

    Science.gov (United States)

    Hesse, Deike; Radloff, Katrin; Jaschke, Alexander; Lagerpusch, Merit; Chung, Bomee; Tailleux, Anne; Staels, Bart; Schürmann, Annette

    2014-01-01

    The liver is a major organ in whole body lipid metabolism and malfunctioning can lead to various diseases including dyslipidemia, fatty liver disease, and type 2 diabetes. Triglycerides and cholesteryl esters are packed in the liver as very low density lipoproteins (VLDLs). Generation of these lipoproteins is initiated in the endoplasmic reticulum and further maturation likely occurs in the Golgi. ADP-ribosylation factor-related protein 1 (ARFRP1) is a small trans-Golgi-associated guanosine triphosphatase (GTPase) that regulates protein sorting and is required for chylomicron lipidation and assembly in the intestine. Here we show that the hepatocyte-specific deletion of Arfrp1 (Arfrp1liv−/−) results in impaired VLDL lipidation leading to reduced plasma triglyceride levels in the fasted state as well as after inhibition of lipoprotein lipase activity by Triton WR-1339. In addition, the concentration of ApoC3 that comprises 40% of protein mass of secreted VLDLs is markedly reduced in the plasma of Arfrp1liv−/− mice but accumulates in the liver accompanied by elevated triglycerides. Fractionation of Arfrp1liv−/− liver homogenates reveals more ApoB48 and a lower concentration of triglycerides in the Golgi compartments than in the corresponding fractions from control livers. In conclusion, ARFRP1 and the Golgi apparatus play an important role in lipoprotein maturation in the liver by influencing lipidation and assembly of proteins to the lipid particles. PMID:24186947

  6. Analysis of the thermal monitoring data collected at the Peach Bottom Atomic Power Station

    International Nuclear Information System (INIS)

    Witten, A.J.; Gray, D.D.

    1977-01-01

    A comprehensive study of the data collected as part of the environmental technical specifications program for Units 2 and 3 of the Peach Bottom Atomic Power Station was conducted for the Office of Nuclear Regulatory Research of the U.S. Nuclear Regulatory Commission. The study included an analysis of both the hydrothermal and ecological data collected from 1967 through 1976. This paper presents the details of the hydrothermal analysis performed under this program. The two primary methods used for temperature monitoring, during both the preoperational and operational periods of the program, are a fixed thermograph network and boat survey measurements. Analysis of the boat survey data provides a fine resolution demonstrating variations in ambient temperature in Conowingo Pond, as well as providing a qualitative picture of the thermal plume produced by the Peach Bottom thermal discharge. The data from 18 thermograph stations was used for a quantitative probability analysis

  7. Study on the viability of peach and apple pollen treated with gamma rays 60Co

    International Nuclear Information System (INIS)

    Filev, K.; Angelov, A.

    1976-01-01

    Pollen from Marygold peach and from Golden Delicious apple varieties was treated with gamma-rays at rates of 0.5, 1.5, 10.0, 50.0, 100, 200, 300, 400 and 500 kR, the irradiation intensity being 1350 r/min. Results point to a different influence of gamma-rays irradiation on the pollen germination. Apple pollen proved more radiosensitive. Least dose of o.5 kR exerted a stimulative effect in the pollen germination. As the irradiation dose rcse, the germination percentage declined. Doses of over 400 kR destroyed completely pollen viability in both fruit species. A linear semi-logarithmical dependence was ascertained between the irradiation rate and pollen germination. 5 and 10 kR proved most suitable for the purposes of the experimental mutagenesis at peach and apple pollen irradiation. (author)

  8. Analysis of containment venting at the Peach Bottom atomic power station

    International Nuclear Information System (INIS)

    Hanson, D.J.; Blackman, H.S.; Nelson, W.R.; Wright, R.E.; Leonard, M.T.; DiSalvo, R.

    1988-01-01

    An analysis of the extent to which containment venting would be effective in preventing or mitigating the consequences of severe accidents has been completed for the Peach Bottom Units 2 and 3 (BWR-4s with Mark I containments). The analysis indicates that the effectiveness of venting in preventing containment overpressurization highly depends on the sequence of the severe accident. Containment venting can be effective for several classes of sequences, including transients with failure of long-term decay heat removal and loss-of-coolant accidents with breaks inside the containment. However, based on draft procedures and equipment in place at the time of the evaluation, containment venting has limited potential for further reducing the risk associated with three severe accident sequences currently identified as important risk contributors at Peach Bottom. Means of improving the potential for risk reduction is identified, but their influence on risk is not analyzed. (orig./HP)

  9. Model with Peach Bottom Turbine trip and thermal-Hydraulic code TRACE V5P3

    International Nuclear Information System (INIS)

    Mesado, C.; Miro, R.; Barrachina, T.; Verdu, G.

    2014-01-01

    This work is the continuation of the work presented previously in the thirty-ninth meeting annual of the Spanish Nuclear society. The semi-automatic translation of the Thermo-hydraulic model TRAC-BF1 Peach Bottom Turbine Trip to TRACE was presented in such work. This article is intended to validate the model obtained in TRACE, why compare the model results result from the translation with the Benchmark results: NEA/OECD BWR Peach Bottom Turbine Trip (PBTT), in particular is of the extreme scenario 2 of exercise 3, in which there is SCRAM in the reactor. Among other data present in the (transitional) Benchmark , are: total power, axial profile of power, pressure Dome, total reactivity and its components. (Author)

  10. Analysis of containment venting at the Peach Bottom Atomic Power Station

    International Nuclear Information System (INIS)

    Hanson, D.J.; Blackman, H.S.; Nelson, W.R.; Wright, R.E.; Leonard, M.T.; DiSalvo, R.

    1986-01-01

    An analysis of the extent to which containment venting would be effective in preventing or mitigating the consequences of severe accidents has been completed for the Peach Bottom Atomic Power Station, Units 2 and 3 (BWR-4s with Mark I containments). The analysis indicates that the effectiveness of venting in preventing containment overpressurization highly depends on the sequence of the severe accident. Containment venting can be effective for several classes of sequences, including transients with failure of long-term decay heat removal and loss-of-coolant accidents with breaks inside the containment. However, based on draft procedures and equipment in place at the time of the evaluation, containment venting has limited potential for further reducing the risk associated with three severe accident sequences currently identified as important risk contributors at Peach Bottom. Means of improving the potential for risk reduction is identified, but their influence on risk is not analyzed

  11. Influence of Inter stock Grafting as a Dwarfing Component on Peach Trees Development and Fruit Quality

    International Nuclear Information System (INIS)

    Hossain, S.; Nasrulhaq, A.; Mizutani, F.

    2006-01-01

    A field experiment was conducted to investigate the interstock grafting effect as a dwarfing component on Peach tree development and fruit quality. The study was made at the Ehime University Experimental farm located in south-eastern Japan during the period 2001-2005. The results of the field experiment indicated that pruned branches weight and flowers number were lower in interstock than in control trees. However, percent fruit set was a little higher in intersrock treated than in control trees. Fruit yield and fruit weight were lower in interstock treated than control trees. Soluble solids content and maturity index were higher in interstock treated trees than control, with an increasing trend in the period from 2001 to 2005, while titratable acidity showed that the livestock grafting is a useful dwarfing component for controlling the size of peach trees and improvement of fruit quality. (author)

  12. Final summary report on the Peach Bottom End-of-Life Program

    International Nuclear Information System (INIS)

    Steward, K.P.

    1978-07-01

    HTGR design methods verifications have been performed under the Peach Bottom End-of-Life Program by comparison of actual with predicted physics, thermal, fission product, and materials behavior in Peach Bottom. These design methods verifications have utilized the data determined from nondestructive fuel and circuit gamma scanning on-site, from laboratory examinations of samples removed from the primary circuit, and from a complementary program of Peach Bottom fuel element postirradiation examinations at Oak Ridge National Laboratory. Fifty-five driver fuel elements were axially gamma scanned to determine fission product distributions for use in burnup calculations, power profile determinations, and fission product release and redistribution studies. The cesium plateout distribution in the primary circuit was subsequently mapped by gamma scanning the ducting at 12 locations, axially traversing 79 steam generator tubes with Cd Te detectors from the water side, and internally scanning two vertical runs of ducting. Component removal involved trepanning of the primary circuit ducting, obtaining access to the steam generator internals, and removing over 100 superheater, evaporator, and economizer tubing samples. During this phase, macroscopic examinations of the steam generator and ducting internals were performed. Subsequent laboratory examinations of removed samples included radiochemical tests to substantiate in-situ gamma scans and to determine 90 Sr distributions; metallurgical tests to evaluate surface films, microstructural changes, and residual mechanical properties; and tritium permeation tests to provide data for improving HTGR tritium release predictions. The condition and metallurgical integrity of all components examined were found to be excellent, with minimal coolant/substrate interaction and entirely acceptable residual mechanical properties. The suitability of the materials used in the construction of the Peach Bottom HTGR was thereby confirmed

  13. Investigation of energy inputs for peach production using sensitivity analysis in Iran

    International Nuclear Information System (INIS)

    Royan, Mahsa; Khojastehpour, Mehdi; Emadi, Bagher; Mobtaker, Hassan Ghasemi

    2012-01-01

    Highlights: ► We investigated energy use and inputs–output relationship in peach production. ► Total energy consumption in peach production was 37536.96 MJ ha −1 . ► Diesel fuel with about (26.32%) was the major energy consumer. ► Energy use efficiency and energy productivity were 0.55 and 0.29 kg MJ −1 . ► The machinery energy was the most significant input affecting the output level. - Abstract: The purpose of this research was to investigate the energy balance between the energy inputs and yield in peach production in Golestan province of Iran as a case study. The results showed that total energy consumption in peach production was 37536.96 MJ ha −1 where the diesel fuel with about (26.32%) was the major energy consumer. The direct energy shared about (50.98%) whereas the indirect energy did (49.02%). Energy use efficiency, energy productivity, specific energy and net energy were 0.55, 0.29 kg MJ −1 , 3.41 MJ kg −1 and −16642.03 MJ ha −1 , respectively. Econometric assessment results revealed that the energy inputs of human labor, machinery, diesel fuel, chemical fertilizers and farm yard manure had significant influence on the yield. The impact of human labor energy (1.36) was found as the highest among the other input parameters. Sensitivity analysis indicated that the MPP value of energy inputs was between −2.8 and 11.31. Also the MPP value of human labor was the highest, followed by diesel fuel and farm yard manure energy inputs, respectively.

  14. Consumption, preferences and habits of purchasing consumers of peaches and nectarines

    Directory of Open Access Journals (Sweden)

    Gener Augusto Penso

    2018-06-01

    Full Text Available Abstract In Brazil, there are few studies and information available on consumer preferences for peaches and nectarines. This lack of information, has hindered some aspects of the production chain, as the choice and exploitation of more appropriate cultivars, offer periods, adoption of marketing and marketing strategies, consequently affecting the availability and commercialization of fruits. This study aimed to analyze the consumer preference of peaches and nectarines, evaluate the traits observed for their acquisition, and the marketplace where they are preferably purchased. The research was carried out from November 2016 to February 2017, using the descriptive research methodology, by Survey method and Web 2.0. A structured questionnaire was applied with closed questions. An online form questionnaire was prepared using the Google docs application and also distributed online, through the internet social Medias. 610 people participated in the survey, with predominance of consumers living in the South and Southeast regions of the country, in the age groups of 21 to 30 years old, with a predominance of female participants. Consumption is mainly in the in natura form, and often predominates certain times a year. Consumers buy these fruits mainly in the supermarkets/hypermarkets. The consumers of peaches and nectarines in Brazil have a higher preference for fruits with yellow flesh and red epidermis, followed by fruits with white flesh and intense red epidermis, both preferably with free stone. The main criteria for the acquisition of peaches and nectarines in Brazil are fruit appearance (presence and/or absence of defects, bruises, etc.; followed by price, color of epidermis and fruit size.

  15. Gas exchanges in peach palms as a function of the spad chlorophyll meter readings

    Directory of Open Access Journals (Sweden)

    Maria Luiza Sant'anna Tucci

    2011-10-01

    Full Text Available The close relationship between the chlorophyll-meters readings and the total chlorophyll and nitrogen contents in leaves, has allowed their evaluation both in annual and perennial species. Besides, some physiological events such as the CO2 assimilation have also been estimated by chlorophyll meters. This work was carried out aiming to evaluate the gas exchanges of peach palms as a function of the chlorophyll SPAD-Meter readings. Three year-old peach palms from Yurimaguas, Peru were studied in Ubatuba, SP, Brazil, spaced 2 x 1 m in area under a natural gradient of organic matter which allowed four plots to be considered, according to the peach palms leaves colors, from light yellow to dark green. The SPAD readings and the stomatal frequency of leaflets were evaluated. The photosynthetic photon flux density (PPFD, μmol m-2 s-1, the leaf temperature (Tleaf, ºC, the CO2 assimilation (A, μmol m-2 s-1, the stomatal conductance (g s, mol m-2 s-1, the transpiration (E, mmol m-2 s-1 and the intercellular CO2 concentration (Ci, μmol mol-1 were evaluated with a portable infrared gas analyzer (LCA-4, ADC BioScientific Ltd., Great Amwell, U.K.. A linear increase in the CO2 assimilation as a function of the SPAD readings (y = -0.34 + 0.19x, R² = 0.99, indicates that they can be a rapid and cheap complementary method to evaluate in peach palms some important physiological events, such as CO2 assimilation.

  16. Supercritical fluid extraction of peach (Prunus persica) almond oil: process yield and extract composition.

    Science.gov (United States)

    Mezzomo, Natália; Mileo, Bruna R; Friedrich, Maria T; Martínez, Julian; Ferreira, Sandra R S

    2010-07-01

    Peach kernels are industrial residues from the peach processing, contain oil with important therapeutic properties and attractive nutritional aspects because of the high concentration of oleic and linoleic acids. The extraction method used to obtain natural compounds from raw matter is critical for product quality definition. Thus, the aim of this work was to compare peach almond extraction yields obtained by different procedures: soxhlet extractions (Sox) with different solvents; hydrodistillation (HD); ethanolic maceration (Mac) followed by fractionation with various solvents, and supercritical fluid extraction (SFE) at 30, 40 and 50 degrees C and at 100, 200 and 300bar, performed with pure CO(2) and with a co-solvent. The extracts were evaluated with respect to fatty acid composition (FAC), fractionated chemical profile (FCP) and total phenolic content (TPC). The Sox total yields were generally higher than those obtained by SFE. The crossover pressure for SFE was between 260 and 280bar. The FAC results show oleic and linoleic acids as main components, especially for Sox and SFE extracts. The FCP for samples obtained by Sox and Mac indicated the presence of benzaldehyde and benzyl alcohol, components responsible for almond flavor and with important industrial uses, whereas the SFE extracts present a high content of a possible flavonoid. The higher TPC values were obtained by Sox and Mac with ethanol. In general, the maximum pressure in SFE produced the highest yield, TPC and oleic acid content. The use of ethanol at 5% as co-solvent in SFE did not result in a significant effect on any evaluated parameter. The production of peach almond oil through all techniques is substantially adequate and SFE presented advantages, with respect to the quality of the extracts due to the high oleic acid content, as presented by some Sox samples. Copyright (c) 2010 Elsevier Ltd. All rights reserved.

  17. HPLC-DAD-ESIMS analysis of phenolic compounds in nectarines, peaches, and plums.

    Science.gov (United States)

    Tomás-Barberán, F A; Gil, M I; Cremin, P; Waterhouse, A L; Hess-Pierce, B; Kader, A A

    2001-10-01

    The phenolic compounds of 25 peach, nectarine, and plum cultivars were studied and quantified by HPLC-DAD-ESIMS. Hydroxycinnamates, procyanidins, flavonols, and anthocyanins were detected and quantified. White and yellow flesh nectarines and peaches, and yellow and red plums, were analyzed at two different maturity stages with consideration of both peel and flesh tissues. HPLC-MS analyses allowed the identification of procyanidin dimers of the B- and A-types, as well as the presence of procyanidin trimers in plums. As a general rule, the peel tissues contained higher amounts of phenolics, and anthocyanins and flavonols were almost exclusively located in this tissue. No clear differences in the phenolic content of nectarines and peaches were detected or between white flesh and yellow flesh cultivars. There was no clear trend in phenolic content with ripening of the different cultivars. Some cultivars, however, had a very high phenolic content. For example, the white flesh nectarine cultivar Brite Pearl (350-460 mg/kg hydroxycinnamates and 430-550 mg/kg procyanidins in flesh) and the yellow flesh cv. Red Jim (180-190 mg/kg hydroxycinnamates and 210-330 mg/kg procyanidins in flesh), contained 10 times more phenolics than cultivars such as Fire Pearl (38-50 mg/kg hydroxycinnamates and 23-30 mg/kg procyanidins in flesh). Among white flesh peaches, cultivars Snow King (300-320 mg/kg hydroxycinnamates and 660-695 mg/kg procyanidins in flesh) and Snow Giant (125-130 mg/kg hydroxycinnamates and 520-540 mg/kg procyanidins in flesh) showed the highest content. The plum cultivars Black Beaut and Angeleno were especially rich in phenolics.

  18. Chromatin-associated regulation of sorbitol synthesis in flower buds of peach.

    Science.gov (United States)

    Lloret, Alba; Martínez-Fuentes, Amparo; Agustí, Manuel; Badenes, María Luisa; Ríos, Gabino

    2017-11-01

    PpeS6PDH gene is postulated to mediate sorbitol synthesis in flower buds of peach concomitantly with specific chromatin modifications. Perennial plants have evolved an adaptive mechanism involving protection of meristems within specialized structures named buds in order to survive low temperatures and water deprivation during winter. A seasonal period of dormancy further improves tolerance of buds to environmental stresses through specific mechanisms poorly known at the molecular level. We have shown that peach PpeS6PDH gene is down-regulated in flower buds after dormancy release, concomitantly with changes in the methylation level at specific lysine residues of histone H3 (H3K27 and H3K4) in the chromatin around the translation start site of the gene. PpeS6PDH encodes a NADPH-dependent sorbitol-6-phosphate dehydrogenase, the key enzyme for biosynthesis of sorbitol. Consistently, sorbitol accumulates in dormant buds showing higher PpeS6PDH expression. Moreover, PpeS6PDH gene expression is affected by cold and water deficit stress. Particularly, its expression is up-regulated by low temperature in buds and leaves, whereas desiccation treatment induces PpeS6PDH in buds and represses the gene in leaves. These data reveal the concurrent participation of chromatin modification mechanisms, transcriptional regulation of PpeS6PDH and sorbitol accumulation in flower buds of peach. In addition to its role as a major translocatable photosynthate in Rosaceae species, sorbitol is a widespread compatible solute and cryoprotectant, which suggests its participation in tolerance to environmental stresses in flower buds of peach.

  19. Impact evaluation of integrated food-bioenergy systems: A comparative LCA of peach nectar

    International Nuclear Information System (INIS)

    De Menna, Fabio; Vittuari, Matteo; Molari, Giovanni

    2015-01-01

    Processed food products present high energy intensity, along with a large amount of food losses and waste. The recovery of residual biomass as integrated renewable energy source could represent an interesting option for the substitution of fossil energy, contributing to the transition of agro-food sector towards a low-carbon economy. Two scenarios were compared, in order to evaluate the impacts of a fossil fuel-based food chain and the potential benefits of the integration of bioenergy production, using peach nectar as case study. In the first scenario, peach nectar is produced, distributed and consumed using fossil energy, while residuals are wasted. In the second scenario, byproducts from the nectar chain are used to produce bioenergy from combustion or anaerobic digestion, which is then consumed to substitute electricity and heat. A comparative life cycle assessment (LCA) based on the same functional unit was performed. Main results show that, in the conventional scenario, most of the damage derives from land use, especially for sugar and glucose production, from the fossil energy consumption of about 15 MJ l −1 , and the related greenhouse gas (GHG) emissions of 0.91 kg CO 2  eq l −1 . Food waste leads to a loss of about 20 kcal l −1 . Bioenergy integration would allow a 13–15% damage reduction, mainly due to the substitution of indirect energy consumption. The effects on human health and ecosystem quality are limited. - Highlights: • Up to 15 MJ l −1 of fossil energy are needed to produce 2.7 MJ of peach nectar. • About 20 out of 648 kcal l −1 of peach and nectar are wasted along the supply chain. • Added ingredients (sugar and glucose) cause a large share of land use impact. • Bioenergy from waste reduces up to 37% of non-renewable energy consumption

  20. Yield and fruit quality of peach scion by using rootstocks propagated by air layering and seed

    OpenAIRE

    Picolotto,Luciano; Fachinello,José Carlos; Bianchi,Valmor João; Manica-Berto,Roberta; Pasa,Mateus da Silveira; Schmitz,Juliano Dutra

    2010-01-01

    In rootstock propagation, several methods can be used; however few practices are adopted due to the lack of information about the behavior of stionic combinations at field. The work aimed to compare the propagation method by air layering with the traditional propagation system by seeds using the scion peach [Prunus persica (L.) Batsch] cultivar Granada. The following variables were measured: yield, fruit weight and size, skin color, flesh firmness, total soluble solids content (TSS) and titat...

  1. An integrated approach for increasing breeding efficiency in apple and peach in Europe.

    Science.gov (United States)

    Laurens, Francois; Aranzana, Maria José; Arus, Pere; Bassi, Daniele; Bink, Marco; Bonany, Joan; Caprera, Andrea; Corelli-Grappadelli, Luca; Costes, Evelyne; Durel, Charles-Eric; Mauroux, Jehan-Baptiste; Muranty, Hélène; Nazzicari, Nelson; Pascal, Thierry; Patocchi, Andrea; Peil, Andreas; Quilot-Turion, Bénédicte; Rossini, Laura; Stella, Alessandra; Troggio, Michela; Velasco, Riccardo; van de Weg, Eric

    2018-01-01

    Despite the availability of whole genome sequences of apple and peach, there has been a considerable gap between genomics and breeding. To bridge the gap, the European Union funded the FruitBreedomics project (March 2011 to August 2015) involving 28 research institutes and private companies. Three complementary approaches were pursued: (i) tool and software development, (ii) deciphering genetic control of main horticultural traits taking into account allelic diversity and (iii) developing plant materials, tools and methodologies for breeders. Decisive breakthroughs were made including the making available of ready-to-go DNA diagnostic tests for Marker Assisted Breeding, development of new, dense SNP arrays in apple and peach, new phenotypic methods for some complex traits, software for gene/QTL discovery on breeding germplasm via Pedigree Based Analysis (PBA). This resulted in the discovery of highly predictive molecular markers for traits of horticultural interest via PBA and via Genome Wide Association Studies (GWAS) on several European genebank collections. FruitBreedomics also developed pre-breeding plant materials in which multiple sources of resistance were pyramided and software that can support breeders in their selection activities. Through FruitBreedomics, significant progresses were made in the field of apple and peach breeding, genetics, genomics and bioinformatics of which advantage will be made by breeders, germplasm curators and scientists. A major part of the data collected during the project has been stored in the FruitBreedomics database and has been made available to the public. This review covers the scientific discoveries made in this major endeavour, and perspective in the apple and peach breeding and genomics in Europe and beyond.

  2. Problem solving capabilities of Peach-fronted Conures (Eupsittula aurea) studied by the string-pulling test

    DEFF Research Database (Denmark)

    Ortiz, Sara Torres; Larsen, Ole Næsbye; Wahlberg, Magnus

    Many studies have indicated advanced cognitive abilities in different species of parrots and ravens. Here we investigated basic cognitive skills of Peach-fronted Conures (Eupsittula aurea) using the string-pulling test. These small Middle-American parrots are often compared to dolphins due...... not be obtained by flying or reached from the ground. By varying different spatial configurations of strings and rewards, different cognitive skills could be investigated. Four conures (two females and two males) were tested. All four individuals solved three out of four tested string configurations (four...... straight strings, two slant strings, two contact-no contact strings) within 14 seconds on average, but all failed in a crossed two-string test. Subsequently we tested the birds with a vertical pulley that required the birds to pull the string down to get the food reward within reach from below...

  3. Comparative Study between Ethanolic and β-Cyclodextrin Assisted Extraction of Polyphenols from Peach Pomace

    Directory of Open Access Journals (Sweden)

    Nada El Darra

    2018-01-01

    Full Text Available Peach byproducts are often regarded as food waste despite their high content in health-promoting components. Amongst the latter, polyphenols are bioactive molecules with significant health benefits. The present study investigated an eco-friendly and cost-effective method using a GRAS food additive, β-cyclodextrin (β-CD, for the recovery of polyphenols from peach pomace. β-CD assisted extraction of polyphenols was compared to that of conventional solvent (ethanol extraction at the same concentrations (10 mg/mL, 20 mg/mL, 30 mg/mL, 40 mg/mL, and 50 mg/mL in terms of quality (antiradical activity and quantity. The extract obtained by 50 mg/mL β-CD assisted extraction showed the highest polyphenol (0.72 mg GAE/g DM and flavonoid (0.35 mg catechin/g of DM concentrations as maximal antiradical activity (6.82% and a noted antibacterial activity. Our results showed the competitiveness of β-CD assisted extraction to recover a high quantity and quality of polyphenols from peach pomace suggesting β-CD as a green alternative method for phenolic extraction.

  4. Repeatability, correlation and path analysis of physical and chemical characteristics of peach fruits

    Directory of Open Access Journals (Sweden)

    Rosana Gonçalves Pires Matias

    2014-12-01

    Full Text Available This study aimed to determine the number of measurements necessary to evaluate physical and chemical characteristics of peach fruits, study the relationships between them and their direct and indirect effects on the content of ascorbic acid and total carotenoids. The characteristics skin and pulp color, fruit weight, suture, equatorial and polar diameters, firmness, soluble solids (SS, titratable acidity (TA, SS/TA ratio, ascorbic acid and total carotenoids were evaluated in 39 cultivars of peach and 3 cultivars of nectarine from the orchard of the Universidade Federal de Viçosa. The repeatability coefficient was estimated by ANOVA and CPCOR. Phenotypic correlation coefficients (rf were estimated and, after the multicollinearity diagnostics, they were unfolded to direct and indirect effects of the explanatory variables on the response variable using path analysis. There was agreement on the magnitude of repeatability coefficients obtained by the two methods; however, they varied among the 14 characteristics. The highest correlations were found between FW, SD, ED and PD. Seven fruits are sufficient to evaluate the physical and chemical characteristics of peach with a correlation coefficient of 90%. The characteristics considered in the path diagrams (b* skin, hº skin, b* pulp, hº pulp, ED, PD, FIR, SS, SS/AT and TC are not the main determinants of the ascorbic acid. The yellow hue of the pulp (hº pulp has the potential to be used in indirect selection for total carotenoids.

  5. Effect of controlled atmosphere on postharvest quality of 'Douradão' peaches

    Directory of Open Access Journals (Sweden)

    Ligia Regina Radomille de Santana

    2011-03-01

    Full Text Available This study was carried out with one of the most important cultivar grown in the State of Sao Paulo, Brazil, which has gained the preference of consumers, due to its sweet taste, intense skin color and large size; however, these fruits are susceptible to chilling injury when cold stored for long periods. The use of controlled atmosphere (CA with elevated CO2 and reduced O2 concentrations prevent the onset of the chilling symptom. Thus, the effect of three different conditions of controlled atmosphere (CA1, CA2, CA3 and Control was evaluated in order to extend the storage life of 'Douradão' peaches. After 14, 21 and 28 days, samples were withdrawn from CA and kept in fresh air at 25 ± 1 °C and 90 ± 5% RH to complete ripening. On the day of removal and after 4 days, were the peaches quality characteristics were evaluated. The results showed that the use of CA during cold storage reduced weight loss and prevented postharvest decay. CA2 and CA3 treatments were effective in keeping good quality of 'Douradão' peaches during 28 days of cold storage, the ripe fruits showed reduced incidence of woolliness, adequate juiciness and flesh firmness. CA1 and Control treatments did not present marketable conditions after 14 days of cold storage.

  6. De novo transcriptome assembly of two different peach cultivars grown in Korea

    Directory of Open Access Journals (Sweden)

    Yeonhwa Jo

    2015-12-01

    Full Text Available Peach (Prunus persica is one of the most popular stone fruits worldwide. Next generation sequencing (NGS has facilitated genome and transcriptome analyses of several stone fruit trees. In this study, we conducted de novo transcriptome analyses of two peach cultivars grown in Korea. Leaves of two cultivars, referred to as Jangtaek and Mibaek, were harvested and used for library preparation. The two prepared libraries were paired-end sequenced by the HiSeq2000 system. We obtained 8.14 GB and 9.62 GB sequence data from Jangtaek and Mibaek (NCBI accession numbers: SRS1056585 and SRS1056587, respectively. The Trinity program was used to assemble two transcriptomes de novo, resulting in 110,477 (Jangtaek and 136,196 (Mibaek transcripts. TransDecoder identified possible coding regions in assembled transcripts. The identified proteins were subjected to BLASTP search against NCBI's non-redundant database for functional annotation. This study provides transcriptome data for two peach cultivars, which might be useful for genetic marker development and comparative transcriptome analyses.

  7. Appraisal of genetic diversity of different peach cultivars and genotypes through rapd markers

    International Nuclear Information System (INIS)

    Bakht, J.; Jamal, N.; Shafi, M

    2012-01-01

    The present study was amid to investigate the genetic diversity of twenty peach cultivars and genotypes by RAPD primers at the Institute of Biotechnology and Genetic Engineering, KPK Agricultural University Peshawar. The result indicated that fifteen primers (GLCO9, GLC20, GLA20, GLA13, GLB10, GLB20, GLB06, GLB19, GLA19, GLB19, GLD16, GLB15, GLA15, GLB12, GLB11) gave genetic distance among the peach cultivars and genotypes under study by PCR amplification. Average genetic diversity (estimated as genetic distance) ranged between 12 and 58%. The molecular size of most of the bands were from 150 bp to 1000 bp. Based on dendrogram analysis, Khyber 1 and Khyber 2 was grouped in cluster A, and Tex-A6-69 and BY-8-135 in cluster B, Candan and 6A were most closely related cultivars and genotypes among the 20 peach cultivars and genotypes while Lering, Flam crest, Tex x-9, early grand and Floradaking were distinctly grouped when compared with the rest of population. (author)

  8. Activated carbon from peach stones using phosphoric acid activation at medium temperatures.

    Science.gov (United States)

    Kim, Dong-Su

    2004-01-01

    In the present study, the activation features of phosphoric acid have been investigated using waste peach stones as the raw material in the production of granular activated carbon. Thermogravimetry/differential thermal analysis was conducted to characterize the thermal behavior of peach stone and titration method was used to evaluate the adsorption capacity of the produced activated carbon. It was observed that the iodine value of the activated carbon increased with activation temperature. However, temperatures higher than 500 degrees C caused a thermal destruction, which resulted in the decrease of the adsorption capacity. Activation longer than 1.5 h at 500 degrees C resulted in thermal degradation of the porous structure of the activated carbon. The adsorption capacity was enhanced with increasing of amounts of phosphoric acid, however, excessive phosphoric acid caused a decrease in the iodine value. In addition, it was found that the carbon yields generally decreased with activation temperature and activation time. Scanning electron microscopy analysis was conducted to observe the changes in the poros structure of the activated carbon produced in different temperatures. Activation of carbon by phosphoric acid was found to be superior to that by CaCl2 and gas activation. The activated carbon produced from peach stone was applied as an adsorbent in the treatment of synthesized wastewater containing cadmium ion and its adsorption capacity was found to be as good as that of the commercial one.

  9. Evaluation of fruit quality, bioactive compounds and total antioxidant activity of flat peach cultivars.

    Science.gov (United States)

    Di Vaio, Claudio; Marallo, Nadia; Graziani, Giulia; Ritieni, Alberto; Di Matteo, Antonio

    2015-08-15

    Fruit quality traits (fresh weight, dry weight, soluble solids content, titratable acidity and firmness) as well as the content of bioactive compounds (phenolic compounds) and total antioxidant activity were evaluated in four commercial cultivars of peach (Greta, Ufo 4, Rome Star and Ufo 6) and four of nectarine (Neve, Planet 1, Maria Carla and Mesembrina) differing in fruit shape (standard or flat) and flesh colour (white or yellow), important cultivars of the Italian and foreign market. The higher fruit organoleptic quality and nutritional profile of flat peach and nectarine cultivars make them candidates for exploiting new market opportunities and the chance to improve profits of farmers. The results showed that assayed quality parameters differed greatly among cultivars. In particular, flesh color and fruit shape accounted for most of the variation in traits underlying organoleptic and nutritional quality. Overall data suggested that the flat white-fleshed nectarine Planet 1, the yellow-fleshed nectarine Mesembrina and the yellow-fleshed peach Ufo 6, because of their profiles in terms of soluble solids content, titratable acidity and bioactive compounds, have the greatest potential to meet current consumer requirements. © 2014 Society of Chemical Industry.

  10. Breeding in peach, cherry and plum: from a tissue culture, genetic, transcriptomic and genomic perspective

    Directory of Open Access Journals (Sweden)

    Basilio Carrasco

    2013-01-01

    Full Text Available This review is an overview of traditional and modern breeding methodologies being used to develop new Prunus cultivars (stone fruits with major emphasis on peach, sweet cherry and Japanese plum. To this end, common breeding tools used to produce seedlings, including in vitro culture tools, are discussed. Additionally, the mechanisms of inheritance of many important agronomical traits are described. Recent advances in stone fruit transcriptomics and genomic resources are providing an understanding of the molecular basis of phenotypic variability as well as the identification of allelic variants and molecular markers. These have potential applications for understanding the genetic diversity of the Prunus species, molecular marker-assisted selection and transgenesis. Simple Sequence Repeat (SSR and Single Nucleotide Polymorphism (SNPs molecular markers are described as useful tools to describe genetic diversity in peach, sweet cherry and Japanese plum. Additionally, the recently sequenced peach genome and the public release of the sweet cherry genome are discussed in terms of their applicability to breeding programs

  11. Nutritional status, yield and composition of peach fruit subjected to the application of organic compost

    Directory of Open Access Journals (Sweden)

    George Wellington Bastos de Melo

    2016-01-01

    Full Text Available The purpose of this study was to evaluate the nutritional state, yield and composition of peaches on peach trees subjected to the application of organic compost to the soil. This experiment was conducted during the 2008 and 2009 cropping season in an orchard containing Chimarrita cultivars grafted onto Capdeboscq rootstocks and Haplumbrept soils in the municipality of Farroupilha (RS, Brazil. The treatments included 0, 9, 18, 36, 72 and 144 liters of organic compost per plant-1 year-1. The total nutrient contents in the leaves, yield components, yields per plant and hectare and compositions of the fruits were evaluated in 2008 and 2009 soon after harvest and after 30 days of storage. The application of organic compost to the soil increased the yield components and the yields per plant and hectare in the two treatments with the highest compost additions, which indicated that the addition of 72 L of compost per plant-1 is ideal economically. The organic compost had little effect on the composition of the peach fruit after harvest and after 30 days of storage.

  12. Fermentation Characteristics and Lactic Acid Bacteria Succession of Total Mixed Ration Silages Formulated with Peach Pomace

    Directory of Open Access Journals (Sweden)

    Xiaodong Hu

    2015-04-01

    Full Text Available The objective of this study was to assess the use of peach pomace in total mixed ration (TMR silages and clarify the differences in aerobic stability between TMR and TMR silages caused by lactic acid bacteria (LAB. The TMR were prepared using peach pomace, alfalfa hay or Leymus chinensis hay, maize meal, soybean meal, cotton meal, limestone, a vitamin-mineral supplement, and salt in a ratio of 6.0:34.0:44.4:7.0:5.0:2.5:1.0:0.1 on a dry matter (DM basis. Fermentation quality, microbial composition, and the predominant LAB were examined during ensiling and aerobic deterioration. The results indicated that the TMR silages with peach pomace were well fermented, with low pH and high lactic acid concentrations. The aerobic stability of TMR silages were significantly higher than that of TMR. Compared with TMR silages with alfalfa hay, TMR silage with Leymus chinensis hay was much more prone to deterioration. Although the dominant LAB were not identical in TMR, the same dominant species, Lactobacillus buchneri and Pediococcus acidilactici, were found in both types of TMR silages after 56 d of ensiling, and they may play an important role in the aerobic stability of TMR silages.

  13. Self-compatible peach (Prunus persica) has mutant versions of the S haplotypes found in self-incompatible Prunus species.

    Science.gov (United States)

    Tao, Ryutaro; Watari, Akiko; Hanada, Toshio; Habu, Tsuyoshi; Yaegaki, Hideaki; Yamaguchi, Masami; Yamane, Hisayo

    2007-01-01

    This study demonstrates that self-compatible (SC) peach has mutant versions of S haplotypes that are present in self-incompatible (SI) Prunus species. All three peach S haplotypes, S (1), S (2), and S (2m), found in this study encode mutated pollen determinants, SFB, while only S (2m) has a mutation that affects the function of the pistil determinant S-RNase. A cysteine residue in the C5 domain of the S (2m)-RNase is substituted by a tyrosine residue, thereby reducing RNase stability. The peach SFB mutations are similar to the SFB mutations found in SC haplotypes of sweet cherry (P. avium) and Japanese apricot (P. mume). SFB (1) of the S (1) haplotype, a mutant version of almond (P. dulcis) S (k) haplotype, encodes truncated SFB due to a 155 bp insertion. SFB (2) of the S (2) and S (2m) haplotypes, both of which are mutant versions of the S (a) haplotype in Japanese plum (P. salicina), encodes a truncated SFB due to a 5 bp insertion. Thus, regardless of the functionality of the pistil determinant, all three peach S haplotypes are SC haplotypes. Our finding that peach has mutant versions of S haplotypes that function in almond and Japanese plum, which are phylogenetically close and remote species, respectively, to peach in the subfamily Prunoideae of the Roasaceae, provides insight into the SC/SI evolution in Prunus. We discuss the significance of SC pollen part mutation in peach with special reference to possible differences in the SI mechanisms between Prunus and Solanaceae.

  14. Extensive in silico analysis of Mimivirus coded Rab GTPase homolog suggests a possible role in virion membrane biogenesis

    Directory of Open Access Journals (Sweden)

    Amrutraj eZade

    2015-09-01

    Full Text Available Rab GTPases are the key regulators of intracellular membrane trafficking in eukaryotes. Many viruses and intracellular bacterial pathogens have evolved to hijack the host Rab GTPase functions, mainly through activators and effector proteins, for their benefit. Acanthamoeba polyphaga mimivirus (APMV is one of the largest viruses and belongs to the monophyletic clade of nucleo-cytoplasmic large DNA viruses (NCLDV. The inner membrane lining is integral to the APMV virion structure. APMV assembly involves extensive host membrane modifications, like vesicle budding and fusion, leading to the formation of a membrane sheet that is incorporated into the virion. Intriguingly, APMV and all group I members of the Mimiviridae family code for a putative Rab GTPase protein. APMV is the first reported virus to code for a Rab GTPase (encoded by R214 gene. Our thorough in silico analysis of the subfamily specific (SF region of Mimiviridae Rab GTPase sequences suggests that they are related to Rab5, a member of the group II Rab GTPases, of lower eukaryotes. Because of their high divergence from the existing three isoforms, A, B and C of the Rab5-family, we suggest that Mimiviridae Rabs constitute a new isoform, Rab5D. Phylogenetic analysis indicated probable horizontal acquisition from a lower eukaryotic ancestor followed by selection and divergence. Furthermore, interaction network analysis suggests that vps34 (a Class III P13K homolog, coded by APMV L615, Atg-8 and dynamin (host proteins are recruited by APMV Rab GTPase during capsid assembly. Based on these observations, we hypothesize that APMV Rab plays a role in the acquisition of inner membrane during virion assembly.

  15. Version VI of the ESTree db: an improved tool for peach transcriptome analysis

    Science.gov (United States)

    Lazzari, Barbara; Caprera, Andrea; Vecchietti, Alberto; Merelli, Ivan; Barale, Francesca; Milanesi, Luciano; Stella, Alessandra; Pozzi, Carlo

    2008-01-01

    Background The ESTree database (db) is a collection of Prunus persica and Prunus dulcis EST sequences that in its current version encompasses 75,404 sequences from 3 almond and 19 peach libraries. Nine peach genotypes and four peach tissues are represented, from four fruit developmental stages. The aim of this work was to implement the already existing ESTree db by adding new sequences and analysis programs. Particular care was given to the implementation of the web interface, that allows querying each of the database features. Results A Perl modular pipeline is the backbone of sequence analysis in the ESTree db project. Outputs obtained during the pipeline steps are automatically arrayed into the fields of a MySQL database. Apart from standard clustering and annotation analyses, version VI of the ESTree db encompasses new tools for tandem repeat identification, annotation against genomic Rosaceae sequences, and positioning on the database of oligomer sequences that were used in a peach microarray study. Furthermore, known protein patterns and motifs were identified by comparison to PROSITE. Based on data retrieved from sequence annotation against the UniProtKB database, a script was prepared to track positions of homologous hits on the GO tree and build statistics on the ontologies distribution in GO functional categories. EST mapping data were also integrated in the database. The PHP-based web interface was upgraded and extended. The aim of the authors was to enable querying the database according to all the biological aspects that can be investigated from the analysis of data available in the ESTree db. This is achieved by allowing multiple searches on logical subsets of sequences that represent different biological situations or features. Conclusions The version VI of ESTree db offers a broad overview on peach gene expression. Sequence analyses results contained in the database, extensively linked to external related resources, represent a large amount of

  16. Performance and characteristics of carcass and non-carcass components of lambs fed peach-palm by-product.

    Science.gov (United States)

    dos Santos Cabral, Ícaro; Azevêdo, José Augusto Gomes; de Almeida, Flávio Moreira; Pereira, Luiz Gustavo Ribeiro; de Araújo, Gherman Garcia Leal; dos Santos Cruz, Cristiane Leal; Nogueira, Abdon Santos; Souza, Lígia Lins; de Oliveira, Gisele Andrade

    2013-11-01

    The objective of this study was to evaluate the effects of supplying the by-product of peach-palm (Bactris gasipaes) on performance and characteristics of carcass and non-carcass components of feedlot lambs. Twenty Santa Ines lambs of 150 days average age and 22.4 ± 3.4 kg body weight were confined in individual pens. A completely randomized design was utilized with four experimental diets composed of: fresh peach-palm by-product enriched with urea, fresh peach-palm by-product + concentrate, silage of peach-palm by-product + concentrate, and silage of peach-palm by-product enriched with 15 % corn meal + concentrate. Intake was evaluated daily, and at the end of 42 days of experiments, lambs were slaughtered and the characteristics of carcass and non-carcass parts were evaluated. Performance and carcass characteristics showed differences between the animals' intake of total mixed rations (TMR) and only the diet with roughage. For the lambs that intaked TMR, the form of utilization of roughage (fresh or as silage) affected animal performance but did not change the carcass characteristics. Dry matter intake and feed conversion were influenced by the form of utilization of the silage (with and without additive). Providing fresh by-product plus concentrate improves lamb performance but does not interfere in the carcass characteristics, compared with the use of by-product in the form of silage.

  17. A one-step multiplex RT-PCR assay for simultaneous detection of four viruses that infect peach.

    Science.gov (United States)

    Yu, Y; Zhao, Z; Jiang, D; Wu, Z; Li, S

    2013-10-01

    A multiplex reverse transcription polymerase chain reaction (mRT-PCR) assay was developed to enable the simultaneous detection and differentiation of four viruses that infect peach, namely Apple chlorotic leaf spot virus (ACLSV), Cherry green ring mottle virus (CGRMV), Prunus necrotic ringspot virus (PNRSV) and Apricot pseudo-chlorotic leaf spot virus (APCLSV). In this study, four pairs of primers, one specific for each virus, were designed; the corresponding PCR products were 632, 439, 346 and 282 bp in length for ACLSV, CGRMV, PNRSV and APCLSV, respectively, and the fragments could be distinguished clearly by agarose gel electrophoresis. The sensitivity and specificity of the method were tested using individual RT-PCR and enzyme-linked immunosorbent assay (ELISA), and the identity of the RT-PCR amplification products was also confirmed by DNA sequencing. The results of RT-PCR and ELISA, along with batch detection using samples collected from peach orchards, revealed that this rapid and simple technique is an effective way to identify the four viruses simultaneously. The mRT-PCR assay described in this study was developed for the simultaneous detection of four peach viruses from infected peach samples is reliable and sensitive. In contrast to conventional uniplex RT-PCR, mRT-PCR is more efficient, reducing costs, time and handling when testing large numbers of samples. This rapid and simple method is useful for large-scale surveys of viruses that infect peach. © 2013 The Society for Applied Microbiology.

  18. Postharvest biological control of brown rot in peaches after cold storage preceded by preharvest chemical control 1

    Directory of Open Access Journals (Sweden)

    Elizandra Pivotto Pavanello

    2015-12-01

    Full Text Available ABSTRACT Pathogenic fungi cause skin darkening and peach quality depreciation in post harvest. Therefore, alternative techniques to chemical treatment are necessary in order to reduce risks to human health. The aim of this study was to evaluate the effect of the application of Trichoderma harzianum in association with different fungicides applied before harvest to 'Eldorado' peaches for brown rot control and other quality parameters during storage. The treatments consisted of five preharvest fungicide applications (control, captan, iprodione, iminoctadine and tebuconazole associated with postharvest application of T. harzianum, after cold storage (with and without application, in three evaluation times (zero, two and four days at 20 °C, resulting in a 5x2x3 factorial design. The application of T. harzianum only brought benefits to the control of brown rot when combined with the fungicide captan, at zero day shelf life. After two days, there was a greater skin darkening in peaches treated with T. harzianum compared with peaches without the treatment, except for peaches treated with the fungicide iprodione and T. harzianum The application of T. harzianum during postharvest showed no benefits for the control of brown rot, however, the association with fungicides reduced the incidence of Rhizopus stolonifer during the shelf life.

  19. Identification of differentially expressed genes associated with changes in the morphology of Pichia fermentans on apple and peach fruit.

    Science.gov (United States)

    Fiori, Stefano; Scherm, Barbara; Liu, Jia; Farrell, Robert; Mannazzu, Ilaria; Budroni, Marilena; Maserti, Bianca E; Wisniewski, Michael E; Migheli, Quirico

    2012-11-01

    Pichia fermentans (strain DISAABA 726) is an effective biocontrol agent against Monilinia fructicola and Botrytis cinerea when inoculated in artificially wounded apple fruit but is an aggressive pathogen when inoculated on wounded peach fruit, causing severe fruit decay. Pichia fermentans grows as budding yeast on apple tissue and exhibits pseudohyphal growth on peach tissue, suggesting that dimorphism may be associated with pathogenicity. Two complementary suppressive subtractive hybridization (SSH) strategies, that is, rapid subtraction hybridization (RaSH) and PCR-based subtraction, were performed to identify genes differentially expressed by P. fermentans after 24-h growth on apple vs. peach fruit. Gene products that were more highly expressed on peach than on apple tissue, or vice versa, were sequenced and compared with available yeast genome sequence databases. Several of the genes more highly expressed, when P. fermentans was grown on peach, were related to stress response, glycolysis, amino acid metabolism, and alcoholic fermentation but surprisingly not to cell wall degrading enzymes such as pectinases or cellulases. The dual activity of P. fermentans as both a biocontrol agent and a pathogen emphasizes the need for a thorough risk analysis of potential antagonists to avoid unpredictable results that could negatively impact the safe use of postharvest biocontrol strategies. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  20. Effect of the addition of peach palm (Bactris gasipaes peel flour on the color and sensory properties of cakes

    Directory of Open Access Journals (Sweden)

    Jader MARTÍNEZ-GIRÓN

    Full Text Available Abstract This study aimed to evaluate the color and sensorial characteristics of a cake made with different amounts of flour made with peach palm peel flour. The flour was added at different concentrations, 2.5, 5.0, 7.5, and 10% (w/w, including a control (tartrazine. Physiochemical analyses were carried out on the cakes, including total carotenoids, CIE-L*a*b* color coordinates, and sensorial attributes. The results demonstrated that the peach palm peel flour substitution increased the total carotenoid content of the cakes. The color test showed that high levels of peach palm peel flour resulted in a decrease in the values of lightness (L* and hue angle (h°, while the values of browning index (BI and color change (ΔE* increased significantly. The qualification of the sensorial evaluation indicated that the quality attributes were acceptable in the cakes made with wheat flour and 7.5% peach palm peel flour. Therefore, it was concluded that flour obtained from peach palm peel is a by-product that can be used as a natural food dye alternative in bread products.

  1. Agronomical Parameters, Sugar Profile and Antioxidant Compounds of “Catherine” Peach Cultivar Influenced by Different Plum Rootstocks

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    Carolina Font i Forcada

    2014-02-01

    Full Text Available The influence of seven plum rootstocks (Adesoto, Monpol, Montizo, Puebla de Soto 67 AD, PM 105 AD, St. Julien GF 655/2 and Constantí 1 on individual and total sugars, as well as on antioxidant content in fruit flesh of “Catherine” peaches, was evaluated for three years. Agronomical and basic fruit quality parameters were also determined. At twelve years after budding, significant differences were found between rootstocks for the different agronomic and fruit quality traits evaluated. The Pollizo plum rootstocks Adesoto and PM 105 AD seem to induce higher sweetness to peach fruits, based on soluble solids content, individual (sucrose, fructose and sorbitol and total sugars. A clear tendency was also observed with the rootstock Adesoto, inducing the highest content of phenolics, flavonoids, vitamin C and relative antioxidant capacity (RAC. Thus, the results of this study demonstrate the significant effect of rootstock on the sugar profile and phytochemical characteristics of peach fruits. In addition, this work shows the importance of the sugar profile, because specific sugars play an important role in peach flavour quality, as well as the studied phytochemical compounds when looking for high quality peaches with enhanced health properties.

  2. RhoGDI: multiple functions in the regulation of Rho family GTPase activities

    DEFF Research Database (Denmark)

    Dovas, Athanassios; Couchman, John R

    2005-01-01

    necessary for the correct targeting and regulation of Rho activities by conferring cues for spatial restriction, guidance and availability to effectors. These potential functions are discussed in the context of RhoGDI-associated multimolecular complexes, the newly emerged shuttling capability...... insight as to how RhoGDI exerts its effects on nucleotide binding, the membrane association-dissociation cycling of the GTPase and how these activities are controlled. Despite the initial negative roles attributed to RhoGDI, recent evidence has come to suggest that it may also act as a positive regulator...... of activities....

  3. Primary coolant chemistry of the Peach Bottom and Fort St. Vrain high-temperature gas-cooled reactors

    International Nuclear Information System (INIS)

    Burnette, R.D.; Baldwin, N.L.

    1980-11-01

    The chemical impurities in the primary coolants of the Peach Bottom and Fort St. Vrain reactors are discussed. The impurity mixtures in the two plants were quite different because the sources of the impurities were different. In the Peach Bottom reactor, the impurities were dominated by H 2 and CH 4 , which are decomposition products of oil. In the Fort St. Vrain reactor, there were high levels of CO, CO 2 , and H 2 O. Although oil ingress at Peach Bottom created carbon deposits on virtually all surfaces, its effect on reactor operation was negligible. Slow outgassing of water from the thermal insulation at Fort St. Vrain caused delays in reactor startup. The overall graphite oxidation in both plants was negligible

  4. Primary coolant chemistry of the Peach Bottom and Fort St. Vrain high temperature gas-cooled reactors

    International Nuclear Information System (INIS)

    Burnette, R.D.; Baldwin, N.L.

    1981-01-01

    The chemical impurities in the primary coolants of the Peach Bottom and Fort St. Vrain reactors are discussed. The impurity mixtures in the two plants were quite different because the sources of the impurities were different. In the Peach Bottom reactor, the impurities were dominated by H 2 and CH 4 , which are decomposition products of oil. In the Fort St. Vrain reactor, there were high levels of CO, CO 2 , and H 2 O. Although oil ingress at Peach Bottom created carbon deposits on virtually all surfaces, its effect on reactor operation was negligible. Slow outgassing of water from the thermal insulation at Fort St. Vrain caused delays in reactor startup. The overall graphite oxidation in both plants was negligible. (author)

  5. NADPH oxidase complex-derived reactive oxygen species, the actin cytoskeleton, and rho GTPases in cell migration

    DEFF Research Database (Denmark)

    Stanley, Alanna; Thompson, Kerry; Hynes, Ailish

    2014-01-01

    Abstract Significance: Rho GTPases are historically known to be central regulators of actin cytoskeleton reorganization. This affects many processes including cell migration. In addition, members of the Rac subfamily are known to be involved in reactive oxygen species (ROS) production through...... mediating cytoskeletal reorganization. Critical Issues: The role of the actin cytoskeleton in providing a scaffold for components of the Nox complex needs to be examined in the light of these new advances. During cell migration, Rho GTPases, ROS, and cytoskeletal organization appear to function as a complex...... compartments. This in conjunction with the analysis of tissues lacking specific Rho GTPases, and Nox components will facilitate a detailed examination of the interactions of these structures with the actin cytoskeleton. In combination with the analysis of ROS production, including its subcellular location...

  6. Liquid chromatography/mass spectrometry investigation of the impact of thermal processing and storage on peach procyanidins.

    Science.gov (United States)

    Hong, Yun-Jeong; Barrett, Diane M; Mitchell, Alyson E

    2004-04-21

    Normal-phase liquid chromatography/mass spectrometry (LC/MS) was used to determine the levels and fate of procyanidins in frozen and canned Ross clingstone peaches as well as in the syrup used in the canning over a 3 month period. Procyanidin oligomers, monomers through undecamers, were identified in Ross clingstone peaches. Optimized methods allowed for the quantitation of oligomers through octamers. The profile of procyanidins in peaches is similar to profiles found in grapes, chocolate, and beverages linked to health benefits such as tea and wine. The monomer content in frozen peeled peaches was found to be 19.59 mg/kg. Dimers (39.59 mg/kg) and trimers (38.81 mg/kg) constituted the largest percent composition of oligomers in the peaches. Tetramers through octamers were present in levels of 17.81, 12.43, 10.62, 3.94 and 1.75 mg/kg, respectively. Thermal processing resulted in an 11% reduction in monomers, a 9% reduction in dimers, a 12% reduction in trimers, a 6% reduction in tetramers, and a 5% reduction in pentamers. Hexamers and heptamers demonstrated an approximate 30% loss, and octamers were no longer detected. Analysis of the syrup after thermal processing indicates that there is a migration of procyanidin monomers through hexamers into the syrup that can account for the losses observed during the canning process. Storage of canned peaches for 3 months demonstrated a time-related loss in higher oligomers and that by 3 months oligomers larger than tetramers are not observed. At 3 months postcanning, levels of monomers had decreased by 10%, dimers by 16%, trimers by 45%, and tetramers by 80%. A similar trend was observed in the canning syrup.

  7. Role of Melatonin in Cell-Wall Disassembly and Chilling Tolerance in Cold-Stored Peach Fruit.

    Science.gov (United States)

    Cao, Shifeng; Bian, Kun; Shi, Liyu; Chung, Hsiao-Hang; Chen, Wei; Yang, Zhenfeng

    2018-06-06

    Melatonin reportedly increases chilling tolerance in postharvest peach fruit during cold storage, but information on its effects on cell-wall disassembly in chilling-injured peaches is limited. In this study, we investigated the role of cell-wall depolymerization in chilling-tolerance induction in melatonin-treated peaches. Treatment with 100 μM melatonin alleviated chilling symptoms (mealiness) characterized by a decrease in fruit firmness and increase in juice extractability in treated peaches during storage. The loss of neutral sugars, such as arabinose and galactose, in both the 1,2-cyclohexylenedinitrilotetraacetic acid (CDTA)- and Na 2 CO 3 -soluble fractions was observed at 7 days in treated peaches, but the contents increased after 28 days of storage. Atomic-force-microscopy (AFM) analysis revealed that the polysaccharide widths in the CDTA- and Na 2 CO 3 -soluble fractions in the treated fruit were mainly distributed in a shorter range, as compared with those in the control fruit. In addition, the expression profiles of a series of cell-wall-related genes showed that melatonin treatment maintained the balance between transcripts of PpPME and PpPG, which accompany the up-regulation of several other genes involved in cell-wall disassembly. Taken together, our results suggested that the reduced mealiness by melatonin was probably associated with its positive regulation of numerous cell-wall-modifying enzymes and proteins; thus, the depolymerization of the cell-wall polysaccharides in the peaches treated with melatonin was maintained, and the treated fruit could soften gradually during cold storage.

  8. Diagnóstico da exposição ocupacional a agrotóxicos na principal região produtora de pêssego para indústria do Brasil Diagnosis of occupational exposure to pesticides in the main growing region of peaches for the industry in Brazil

    Directory of Open Access Journals (Sweden)

    Crislaine Alves Barcellos de Lima

    2009-06-01

    Full Text Available A cultura do pessegueiro é bastante suscetível ao ataque de insetos praga e doenças. Para evitar prejuízos na produção, os persicultores utilizam agrotóxicos. Os objetivos deste trabalho foram caracterizar as propriedades e obter dados para melhor diagnosticar a realidade sobre a exposição ocupacional a agrotóxicos na cultura do pessegueiro. O estudo baseou-se em entrevistas espontâneas, realizadas nas propriedades rurais, junto a 135 persicultores. Os resultados mostram que, de modo geral, a cultura do pessegueiro é praticada em pequenas propriedades, onde a mão-de-obra predominante é a familiar. A aquisição dos agrotóxicos é feita junto a lojas credenciadas, porém, sem receituário agronômico. Entre os agrotóxicos citados, destacam-se o fungicida Carbendazin (Derosal 500 e o inseticida (Perfekthion, que não são registrados para a cultura do pessegueiro. A utilização de equipamento de proteção individual (EPI padrão é uma prática incomum entre os aplicadores.The peach culture is extremely susceptible to the attack of pests and diseases. Aiming to avoid yield losses, peach growers apply pesticides. The objective of this research was to characterize some farms and obtain data to better diagnose the reality about the occupational exposition to pesticides in the peach culture. The methodology was based on spontaneous interviews carried out on farms with 135 peach growers. The results show that in general the peach culture is practiced on small farms where the predominant labor comes from the family. The purchase of agrochemicals is made via accredited shops, but without an agronomic prescription. Among the cited pesticides, the fungicide Carbendazim (Derosal 500 and insecticide (Perfekthion are not labeled in Brazil for the peach culture. The use of personal protective equipment (PPE standard is an uncommon practice among applicators.

  9. Genomic characterization of putative allergen genes in peach/almond and their synteny with apple

    Science.gov (United States)

    Chen, Lin; Zhang, Shuiming; Illa, Eudald; Song, Lijuan; Wu, Shandong; Howad, Werner; Arús, Pere; Weg, Eric van de; Chen, Kunsong; Gao, Zhongshan

    2008-01-01

    Background Fruits from several species of the Rosaceae family are reported to cause allergic reactions in certain populations. The allergens identified belong to mainly four protein families: pathogenesis related 10 proteins, thaumatin-like proteins, lipid transfer proteins and profilins. These families of putative allergen genes in apple (Mal d 1 to 4) have been mapped on linkage maps and subsequent genetic study on allelic diversity and hypoallergenic traits has been carried out recently. In peach (Prunus persica), these allergen gene families are denoted as Pru p 1 to 4 and for almond (Prunus dulcis)Pru du 1 to 4. Genetic analysis using current molecular tools may be helpful to establish the cause of allergenicity differences observed among different peach cultivars. This study was to characterize putative peach allergen genes for their genomic sequences and linkage map positions, and to compare them with previously characterized homologous genes in apple (Malus domestica). Results Eight Pru p/du 1 genes were identified, four of which were new. All the Pru p/du 1 genes were mapped in a single bin on the top of linkage group 1 (G1). Five Pru p/du 2 genes were mapped on four different linkage groups, two very similar Pru p/du 2.01 genes (A and B) were on G3, Pru p/du 2.02 on G7,Pru p/du 2.03 on G8 and Pru p/du 2.04 on G1. There were differences in the intron and exon structure in these Pru p/du 2 genes and in their amino acid composition. Three Pru p/du 3 genes (3.01–3.03) containing an intron and a mini exon of 10 nt were mapped in a cluster on G6. Two Pru p/du 4 genes (Pru p/du 4.01 and 4.02) were located on G1 and G7, respectively. The Pru p/du 1 cluster on G1 aligned to the Mal d 1 clusters on LG16; Pru p/du 2.01A and B on G3 to Mal d 2.01A and B on LG9; the Pru p/du 3 cluster on G6 to Mal d 3.01 on LG12; Pru p/du 4.01 on G1 to Mal d 4.03 on LG2; and Pru p/du 4.02 on G7 to Mal d 4.02 on LG2. Conclusion A total of 18 putative peach/almond allergen genes have

  10. Genomic characterization of putative allergen genes in peach/almond and their synteny with apple

    Directory of Open Access Journals (Sweden)

    Weg Eric

    2008-11-01

    Full Text Available Abstract Background Fruits from several species of the Rosaceae family are reported to cause allergic reactions in certain populations. The allergens identified belong to mainly four protein families: pathogenesis related 10 proteins, thaumatin-like proteins, lipid transfer proteins and profilins. These families of putative allergen genes in apple (Mal d 1 to 4 have been mapped on linkage maps and subsequent genetic study on allelic diversity and hypoallergenic traits has been carried out recently. In peach (Prunus persica, these allergen gene families are denoted as Pru p 1 to 4 and for almond (Prunus dulcisPru du 1 to 4. Genetic analysis using current molecular tools may be helpful to establish the cause of allergenicity differences observed among different peach cultivars. This study was to characterize putative peach allergen genes for their genomic sequences and linkage map positions, and to compare them with previously characterized homologous genes in apple (Malus domestica. Results Eight Pru p/du 1 genes were identified, four of which were new. All the Pru p/du 1 genes were mapped in a single bin on the top of linkage group 1 (G1. Five Pru p/du 2 genes were mapped on four different linkage groups, two very similar Pru p/du 2.01 genes (A and B were on G3, Pru p/du 2.02 on G7,Pru p/du 2.03 on G8 and Pru p/du 2.04 on G1. There were differences in the intron and exon structure in these Pru p/du 2 genes and in their amino acid composition. Three Pru p/du 3 genes (3.01–3.03 containing an intron and a mini exon of 10 nt were mapped in a cluster on G6. Two Pru p/du 4 genes (Pru p/du 4.01 and 4.02 were located on G1 and G7, respectively. The Pru p/du 1 cluster on G1 aligned to the Mal d 1 clusters on LG16; Pru p/du 2.01A and B on G3 to Mal d 2.01A and B on LG9; the Pru p/du 3 cluster on G6 to Mal d 3.01 on LG12; Pru p/du 4.01 on G1 to Mal d 4.03 on LG2; and Pru p/du 4.02 on G7 to Mal d 4.02 on LG2. Conclusion A total of 18 putative peach

  11. The monomeric GTPase RabA2 is required for progression and maintenance of membrane integrity of infection threads during root nodule symbiosis.

    Science.gov (United States)

    Dalla Via, Virginia; Traubenik, Soledad; Rivero, Claudio; Aguilar, O Mario; Zanetti, María Eugenia; Blanco, Flavio Antonio

    2017-04-01

    Progression of the infection canal that conducts rhizobia to the nodule primordium requires a functional Rab GTPase located in Golgi/trans-Golgi that also participate in root hair polar growth. Common bean (Phaseolus vulgaris) symbiotically associates with its partner Rhizobium etli, resulting in the formation of root nitrogen-fixing nodules. Compatible bacteria can reach cortical cells in a tightly regulated infection process, in which the specific recognition of signal molecules is a key step to select the symbiotic partner. In this work, we show that RabA2, a monomeric GTPase from common bean, is required for the progression of the infection canal, referred to as the infection thread (IT), toward the cortical cells. Expression of miss-regulated mutant variants of RabA2 resulted in an increased number of abortive infection events, including bursting of ITs and a reduction in the number of nodules. Nodules formed in these plants were small and contained infected cells with disrupted symbiosome membranes, indicating either early senescence of these cells or defects in the formation of the symbiosome membrane during bacterial release. RabA2 localized to mobile vesicles around the IT, but mutations that affect GTP hydrolysis or GTP/GDP exchange modified this localization. Colocalization of RabA2 with ArfA1 and a Golgi marker indicates that RabA2 localizes in Golgi stacks and the trans-Golgi network. Our results suggest that RabA2 is part of the vesicle transport events required to maintain the integrity of the membrane during IT progression.

  12. Secretion of soluble vascular endothelial growth factor receptor 1 (sVEGFR1/sFlt1 requires Arf1, Arf6, and Rab11 GTPases.

    Directory of Open Access Journals (Sweden)

    Jae-Joon Jung

    Full Text Available The soluble form of vascular endothelial growth factor receptor 1 (sVEGFR-1/sFlt1 is generated by alternative splicing of the FLT1 gene. Secretion of sFlt1 from endothelial cells plays an important role in blood vessel sprouting and morphogenesis. However, excess sFlt1 secretion is associated with diseases such as preeclampsia and chronic kidney disease. To date, the secretory transport process involved in the secretion of sFlt1 is poorly understood. In the present study, we investigated the itinerary of sFlt1 trafficking along the secretory pathway. To understand the timecourse of sFlt1 secretion, endothelial cells stably expressing sFlt1 were metabolically radiolabeled with [(35S]-methionine and cysteine. Our results indicate that after initial synthesis the levels of secreted [(35S]-sFlt1 in the extracellular medium peaks at 8 hours. Treatment with brefeldin A (BFA, a drug which blocks trafficking between the endoplasmic reticulum (ER and the Golgi complex, inhibited extracellular release of sFlt1 suggesting that ER to Golgi and intra-Golgi trafficking of sFlt1 are essential for its secretion. Furthermore, we show that ectopic expression of dominant-negative mutant forms of Arf1, Arf6, and Rab11 as well as siRNA-mediated knockdown of these GTPases block secretion of sFlt1 during normoxic and hypoxic conditions suggesting role for these small GTPases. This work is the first to report role of regulatory proteins involved in sFlt1 trafficking along the secretory pathway and may provide insights and new molecular targets for the modulation of sFlt-1 release during physiological and pathological conditions.

  13. Differential regulation of the Rac1 GTPase-activating protein (GAP) BCR during oxygen/glucose deprivation in hippocampal and cortical neurons.

    Science.gov (United States)

    Smith, Katharine R; Rajgor, Dipen; Hanley, Jonathan G

    2017-12-08

    Brain ischemia causes oxygen and glucose deprivation (OGD) in neurons, triggering a cascade of events leading to synaptic accumulation of glutamate. Excessive activation of glutamate receptors causes excitotoxicity and delayed cell death in vulnerable neurons. Following global cerebral ischemia, hippocampal CA1 pyramidal neurons are more vulnerable to injury than their cortical counterparts, but the mechanisms that underlie this difference are unclear. Signaling via Rho-family small GTPases, their upstream guanine nucleotide exchange factors, and GTPase-activating proteins (GAPs) is differentially dysregulated in response to OGD/ischemia in hippocampal and cortical neurons. Increased Rac1 activity caused by OGD/ischemia contributes to neuronal death in hippocampal neurons via diverse effects on NADPH oxidase activity and dendritic spine morphology. The Rac1 guanine nucleotide exchange factor Tiam1 mediates an OGD-induced increase in Rac1 activity in hippocampal neurons; however, the identity of an antagonistic GAP remains elusive. Here we show that the Rac1 GAP breakpoint cluster region (BCR) associates with NMDA receptors (NMDARs) along with Tiam1 and that this protein complex is more abundant in hippocampal compared with cortical neurons. Although total BCR is similar in the two neuronal types, BCR is more active in hippocampal compared with cortical neurons. OGD causes an NMDAR- and Ca 2+ -permeable AMPAR-dependent deactivation of BCR in hippocampal but not cortical neurons. BCR knockdown occludes OGD-induced Rac1 activation in hippocampal neurons. Furthermore, disrupting the Tiam1-NMDAR interaction with a fragment of Tiam1 blocks OGD-induced Tiam1 activation but has no effect on the deactivation of BCR. This work identifies BCR as a critical player in Rac1 regulation during OGD in hippocampal neurons. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  14. Validation of the Engineering Plant Analyzer methodology with Peach Bottom 2 stability tests

    International Nuclear Information System (INIS)

    Rohatgi, U.S.; Mallen, A.N.; Cheng, H.S.; Wulff, W.

    1994-01-01

    The Engineering Plant Analyzer (EPA) had been developed in 1984 at Brookhaven National Laboratory to simulate plant transients in boiling water reactors (BWR). Recently, the EPA with its High-Speed Interactive Plant Analyzer code for BWRs ( ppercase HIPA-BWR ) simulated for the first time oscillatory transients with large, non-linear power and flow amplitudes; transients which are centered around the March 9, 1988 instability at the LaSalle-2 BWR power plant.The EPA's capability to simulate oscillatory transients has been demonstrated first by comparing simulation results with LaSalle-2 plant data (Wulff et al., NUREG/CR-5816, BNL-NUREG-52312, Brookhaven National Laboratory, 1992). This paper presents an EPA assessment on the basis of the Peach Bottom 2 instability tests (Carmichael and Niemi, EPRI NP-564, Electric Power Research Institute, Palo Alto, CA, 1978). This assessment of the EPA appears to constitute the first validation of a time-domain reactor systems code on the basis of frequency-domain criteria, namely power spectral density, gain and phase shift of the pressure-to-power transfer function.The reactor system pressure was disturbed in the Peach Bottom 2 power plant tests, and in their EPA simulation, by a pseudo-random, binary sequence signal. The data comparison revealed that the EPA predicted for Peach Bottom tests PT1, PT2, and PT4 the gain of the power-to-pressure transfer function with the biases and standard deviations of (-10±28)%, (-1±40)% and (+28±52)%, respectively. The respective frequencies at the peak gains were predicted with the errors of +6%, +3%, and -28%. The differences between the predicted and the measured phase shift increased with increasing frequency, but stayed within the margin of experimental uncertainty. ((orig.))

  15. Effect of regulated deficit irrigation and crop load on the antioxidant compounds of peaches.

    Science.gov (United States)

    Buendía, Begoña; Allende, Ana; Nicolás, Emilio; Alarcón, Juan J; Gil, Maria I

    2008-05-28

    The use of regulated deficit irrigation (RDI) strategies is becoming a common practice in areas with low water availability. Little information is available about the effects of RDI on the antioxidant content of fruits. In this study, the influence of RDI on the content of vitamin C, phenolic compounds and carotenoids was investigated. Two irrigation strategies, fully irrigated (FI) and RDI, were compared at two levels of thinning, commercial and half of the commercial crop load. RDI strategies affected the content of vitamin C, phenolics and carotenoids of Flordastar peaches. RDI caused fruit peel stress lowering the content of vitamin C and carotenoids, while increasing the phenolic content, mainly anthocyanins and procyanidins. Fruit weight was the only quality index influenced by the crop load as it increased in FI fruits at low crop load. In general, fruits from commercial crop load had slightly higher content of antioxidants to fruits from low crop load, although these influences were only observed in the peel. Additionally, the influence of irrigation controlled by two sensors related to plant water level, maximum daily trunk shrinkage (MDS) and sap flow (SF) on the antioxidant constituents of peaches was evaluated. The response of the fruits to SF sensor was similar to that observed for RDI strategy. According to the tested water sensors, SF did not act as a good plant-based water indicator for use in irrigation scheduling, as it caused an increase in the content of phenolics, similar to that observed for fruits subjected to RDI. Therefore, selection of RDI strategies and plant water indicators should be taken into account as they affect the content of antioxidants of peaches.

  16. Phytosanitary irradiation of peach fruit moth (Lepidoptera: Carposinidae) in apple fruits

    International Nuclear Information System (INIS)

    Zhan, Guoping; Li, Baishu; Gao, Meixu; Liu, Bo; Wang, Yuejin; Liu, Tao; Ren, Lili

    2014-01-01

    Peach fruit moth, Carposina sasakii Matsumura, is a serious pest of many pome and stone fruits and presents a quarantine problem in some export markets. It is widely distributed in pome fruit production areas in China, Japan, Korea, North Korea and the Far Eastern Federal District of Russia. In this investigation, gamma radiation dose–response tests were conducted with late eggs (5-d-old) and various larval stages, followed by large-scale confirmatory tests on the most tolerant stage in fruit, the fifth instar. The dose-response tests, with the target radiation dose of 20 (late eggs), 40, 60, 80, 100, 120, 140, and 160 Gy (late fifth instars in vitro) respectively applied to all stages, showed that the tolerance to radiation increased with increasing age and developmental stage. The fifth instar (most advanced instar in fruits) was determined to be the most tolerant stage requiring an estimated minimum absorbed dose of 208.6 Gy (95% CI: 195.0, 226.5 Gy) to prevent adult emergence at 99.9968% efficacy (95% confidence level). In the confirmatory tests, irradiation was applied to 30,850 late fifth instars in apple fruits with a target dose of 200 Gy (171.6–227.8 Gy measured), but only 4 deformed adults emerged that died 2 d afterwards without laying eggs. A dose of 228 Gy may be recommended as a phytosanitary irradiation treatment under ambient atmosphere for the control of peach fruit moth on all commodities with an efficacy of 99.9902% at 95% confidence level. - Highlights: • Dose–response tests were conducted on eggs and all larval stages. • Fifth instar is the most tolerant stage that could be shipped in fruits. • None normal-looking adult emerged from 30,850 fifth instars in confirmatory tests. • A minimum of 228 Gy is suggested for phytosanitary irradiation of peach fruit moth

  17. Effect of Water Deficit Stress on Peach Growth under Commercial Orchard Management Conditions

    Directory of Open Access Journals (Sweden)

    M. Rahmati

    2015-06-01

    Full Text Available In order to study the sensitivity of vegetative growth to water deficit stress of a late-maturing peach (Prunus persica L. cv. Elberta under orchard conditions, an experiment was conducted as randomized complete-block design with three treatments and four repetitions in Shahdiran commercial orchard in Mashhad during 2011. Three irrigation treatments including 360 (low stress, 180 (moderate stress and 90 (severe stress m3ha-1week-1 using a drip irrigation system (minimum stem water potential near harvest: -1.2, -1.5 and -1.7 MPa, respectively from the mid-pit hardening stage (12th of June until harvest (23rd of Sep. applied. Predawn, stem and leaf water potentials, leaf photosynthesis, transpiration, stomatal conductance and leaf temperature, the number of new shoots on fruit bearing shoots and vegetative shoots lengths during growing season as well as leaf area at harvest were measured. The results showed that water deficit stress had negative effects on peach tree water status, thereby resulting in decreased leaf gas exchange and tree vegetative growth. As significant decreased assimilate production of tree was resulted from both decreased leaf assimilation rate (until about 23 % and 50 %, respectively under moderate and severe stress conditions compared to low stress conditions and decreased leaf area of tree (until about 57% and 79%, respectively under moderate and severe stress conditions compared to low stress conditions at harvest. The significant positive correlation between leaf water potential and vegetative growth of peach revealed that shoot growth would decrease by 30% and 50% of maximum at leaf water potential of –1.56 and –2.30 MPa, respectively.

  18. Peach Bottom Turbine Trip Simulations with RETRAN Using INER/TPC BWR Transient Analysis Method

    International Nuclear Information System (INIS)

    Kao Lainsu; Chiang, Show-Chyuan

    2005-01-01

    The work described in this paper is benchmark calculations of pressurization transient turbine trip tests performed at the Peach Bottom boiling water reactor (BWR). It is part of an overall effort in providing qualification basis for the INER/TPC BWR transient analysis method developed for the Kuosheng and Chinshan plants. The method primarily utilizes an advanced system thermal hydraulics code, RETRAN02/MOD5, for transient safety analyses. Since pressurization transients would result in a strong coupling effect between core neutronic and system thermal hydraulics responses, the INER/TPC method employs the one-dimensional kinetic model in RETRAN with a cross-section data library generated by the Studsvik-CMS code package for the transient calculations. The Peach Bottom Turbine Trip (PBTT) tests, including TT1, TT2, and TT3, have been successfully performed in the plant and assigned as standards commonly for licensing method qualifications for years. It is an essential requirement for licensing purposes to verify integral capabilities and accuracies of the codes and models of the INER/TPC method in simulating such pressurization transients. Specific Peach Bottom plant models, including both neutronics and thermal hydraulics, are developed using modeling approaches and experiences generally adopted in the INER/TPC method. Important model assumptions in RETRAN for the PBTT test simulations are described in this paper. Simulation calculations are performed with best-estimated initial and boundary conditions obtained from plant test measurements. The calculation results presented in this paper demonstrate that the INER/TPC method is capable of calculating accurately the core and system transient behaviors of the tests. Excellent agreement, both in trends and magnitudes between the RETRAN calculation results and the PBTT measurements, shows reliable qualifications of the codes/users/models involved in the method. The RETRAN calculated peak neutron fluxes of the PBTT

  19. Rho GTPase activity modulates paramyxovirus fusion protein-mediated cell-cell fusion

    International Nuclear Information System (INIS)

    Schowalter, Rachel M.; Wurth, Mark A.; Aguilar, Hector C.; Lee, Benhur; Moncman, Carole L.; McCann, Richard O.; Dutch, Rebecca Ellis

    2006-01-01

    The paramyxovirus fusion protein (F) promotes fusion of the viral envelope with the plasma membrane of target cells as well as cell-cell fusion. The plasma membrane is closely associated with the actin cytoskeleton, but the role of actin dynamics in paramyxovirus F-mediated membrane fusion is unclear. We examined cell-cell fusion promoted by two different paramyxovirus F proteins in three cell types in the presence of constitutively active Rho family GTPases, major cellular coordinators of actin dynamics. Reporter gene and syncytia assays demonstrated that expression of either Rac1 V12 or Cdc42 V12 could increase cell-cell fusion promoted by the Hendra or SV5 glycoproteins, though the effect was dependent on the cell type expressing the viral glycoproteins. In contrast, RhoA L63 decreased cell-cell fusion promoted by Hendra glycoproteins but had little affect on SV5 F-mediated fusion. Also, data suggested that GTPase activation in the viral glycoprotein-containing cell was primarily responsible for changes in fusion. Additionally, we found that activated Cdc42 promoted nuclear rearrangement in syncytia

  20. Enhanced accumulation of atropine in Atropa belladonna transformed by Rac GTPase gene isolated from Scoparia dulcis.

    Science.gov (United States)

    Asano, Kyouhei; Lee, Jung-Bum; Yamamura, Yoshimi; Kurosaki, Fumiya

    2013-12-01

    Leaf tissues of Atropa belladonna were transformed by Sdrac2, a Rac GTPase gene, that is isolated from Scoparia dulcis, and the change in atropine concentration of the transformants was examined. Re-differentiated A. belladonna overexpressing Sdrac2 accumulated considerable concentration of atropine in the leaf tissues, whereas the leaves of plants transformed by an empty vector accumulated only a very low concentration of the compound. A. belladonna transformed by CASdrac2, a modified Sdrac2 of which translate was expected to bind guanosine triphosphate (GTP) permanently, accumulated very high concentrations of atropine (approximately 2.4-fold excess to those found in the wild-type plant in its natural habitat). In sharp contrast, the atropine concentration in transformed A. belladonna prepared with negatively modified Sdrac2, DNSdrac2, expected to bind guanosine diphosphate instead of GTP, was very low. These results suggested that Rac GTPases play an important role in the regulation of secondary metabolism in plant cells and that overexpression of the gene(s) may be capable of enhancing the production of natural products accumulated in higher plant cells.

  1. The Rho-GTPase binding protein IQGAP2 is required for the glomerular filtration barrier.

    Science.gov (United States)

    Sugano, Yuya; Lindenmeyer, Maja T; Auberger, Ines; Ziegler, Urs; Segerer, Stephan; Cohen, Clemens D; Neuhauss, Stephan C F; Loffing, Johannes

    2015-11-01

    Podocyte dysfunction impairs the size selectivity of the glomerular filter, leading to proteinuria, hypoalbuminuria, and edema, clinically defined as nephrotic syndrome. Hereditary forms of nephrotic syndrome are linked to mutations in podocyte-specific genes. To identify genes contributing to podocyte dysfunction in acquired nephrotic syndrome, we studied human glomerular gene expression data sets for glomerular-enriched gene transcripts differentially regulated between pretransplant biopsy samples and biopsies from patients with nephrotic syndrome. Candidate genes were screened by in situ hybridization for expression in the zebrafish pronephros, an easy-to-use in vivo assay system to assess podocyte function. One glomerulus-enriched product was the Rho-GTPase binding protein, IQGAP2. Immunohistochemistry found a strong presence of IQGAP2 in normal human and zebrafish podocytes. In zebrafish larvae, morpholino-based knockdown of iqgap2 caused a mild foot process effacement of zebrafish podocytes and a cystic dilation of the urinary space of Bowman's capsule upon onset of urinary filtration. Moreover, the glomerulus of zebrafish morphants showed a glomerular permeability for injected high-molecular-weight dextrans, indicating an impaired size selectivity of the glomerular filter. Thus, IQGAP2 is a Rho-GTPase binding protein, highly abundant in human and zebrafish podocytes, which controls normal podocyte structure and function as evidenced in the zebrafish pronephros.

  2. Regulation of Rac1 GTPase activity by quinine through G-protein and bitter taste receptor T2R4.

    Science.gov (United States)

    Sidhu, Crystal; Jaggupilli, Appalaraju; Chelikani, Prashen; Bhullar, Rajinder P

    2017-02-01

    Rac1 belongs to the Rho family of small GTPases and regulates actin cytoskeleton reorganization. T2R4 is a bitter taste receptor belonging to the G protein-coupled receptor family of proteins. In addition to mediating bitter taste perception from the tongue, T2R4s are found in extra-oral tissues, e.g., nasal epithelium, airways, brain, testis suggesting a much broader physiological function for these receptors. Anti-malarial drug and a bitter tasting compound, quinine, is a known agonist for T2R4, whereas BCML (Nα,Nα-Bis(carboxymethyl)-L-lysine) acts as an inverse agonist. Using western blot and Ca ++ mobilization assays, the effects of quinine on Rac1 activity in HEK293T cells stably expressing T2R4/Gα 16/44 , T2R4, or Gα 16/44 and transiently transfected with HA-Rac1 were investigated. Quinine treatment caused a significant reduction in the amount of active Rac1, whereas in the presence of BCML, quinine failed to cause any significant change in active Rac1. No significant change in Rac1 activity was observed in BAPTA-AM plus quinine-treated Gα 16/44 cells, suggesting possibility of a pathway in addition to the canonical Ca ++ -dependent pathway. A noticeable role for Gα 16/44 independent of T2R4 is observed in quinine-mediated Rac1 inactivation. Further, a significant difference in quinine-induced Ca ++ response in T2R4/Gα 16/44 or T2R4 cells was observed validating the partial role of calcium and importance of Gα 16/44 . This study is the first to show an inhibitory downstream action of a T2R4 agonist on Rac1 function. Further investigation will help in better understanding the downstream signal transduction network of T2R4 and its extra-oral physiological roles.

  3. Ohmic heating of peaches in the wide range of frequencies (50 Hz to 1 MHz).

    Science.gov (United States)

    Shynkaryk, Mykola V; Ji, Taehyun; Alvarez, Valente B; Sastry, Sudhir K

    2010-09-01

    The ohmic heating (OH) rate of peaches was studied at fixed electric field strength of 60 V.cm⁻¹, square-shaped instant reversal bipolar pulses, and frequencies varying within 50 Hz to 1 MHz. Thermal damage of tissue was evaluated from electrical admittivity. It showed that the time for half disruption (τ(T)) of tissue was required more than 10 h at temperatures below 40 °C. However, cellular thermal disruption occurred almost instantly (τ(T) 90 °C). Electrical conductivity σ(o) and admittivity σ(o)* of tissue at T(o)= 0 °C and their temperature coefficients (m, m*) were calculated. For freeze-thawed tissues, σ and σ* as well as m and m* were nearly indifferent to the frequency. However, for the intact tissue, both σ(o), σ(o)* and m, m* were frequency dependent. For freeze-thawed product, the power factor (P) was approximately equal to 1 and indifferent to the frequency and temperature. On the other hand, strong frequency dependence was observed for intact tissue with the minimum P approximately equal to 0.68 in the range of tens of kHz. The time required to reach a target temperature t(f) was evaluated. The t(f) increased with frequency up to the middle of the range of tens of kHz and thereafter continuously decreased. Samples exposed to the low-frequency electric field demonstrated faster electro-thermal damage rates. The textural relaxation data supported more intense damage kinetics at low-frequency OH. It has been demonstrated that a combination of high-frequency OH with pasteurization at moderate temperature followed by rapid cooling minimizes texture degradation of peach tissue. In this study, we investigated the electric field frequency effect on the rate of OH of peaches. It was shown that the time required for reaching the target temperature is strongly dependent upon the frequency. Samples exposed to low-frequency OH demonstrated higher electro-thermal damage rates. It has been shown that the combination of high-frequency OH with

  4. [Effect of supplementation with peach palm as source of vitamin A: study with rats].

    Science.gov (United States)

    Yuyama, L K; Cozzolino, S M

    1996-02-01

    The effect of supplementation with peach palm (Bactris gasipaes H.B.K.), as an alternative source of vit. A, in the regional diet of Manaus, AM, Brazil, in which the pulp was cooked and transformed into flour, was studied. The biological trial involved rats which were depleted in zinc and vitamin A, followed by repletion using the regional diet (RD), RD plus peach palm and RD plus vitamin A. The parameters used to determine the utilization of vitamin A were the vitamin A concentrations in the liver and plasma, and the growth of the animals. The diet was prepared according to the data of Shrimpton and Giugliano for families earning less than two legal minimum salaries. Adult post-partum rats were used, with six male pups each, which received a diet based on casein washed with 1% EDTA, without the addition of zinc or vitamin A for a period of 25 days, for the purpose of obtaining newly-weaned animals which were deficient in Zn and Vit.A. A control group received a diet also based on casein washed with 1% EDTA, but with all the nutrients in the quantities suggested by the Committee on Laboratory Animal Diets. The repletion period of the newly-weaned rats was of 30 days and the experimental design was entirely randomized with four groups of eight rats each. The diet supplementation followed the recommendations of the Committee on Laboratory Animal Diets. At the end of the experiment, it was observed that rats which consumed the diet based on the regional diet of Manaus supplemented with either peach palm or vitamin A showed a significantly greater concentration of vitamin A in the liver, 43.3 +/- 6.5 micrograms/g, 42.0 +/- 4.3 micrograms/g, respectively in relation to the regional diet, 5.5 +/- 1.1 micrograms/g (p < 0.05). The amount of zinc present in the regional diet, 10.7 mg per day, was bioavailable as determined by the concentration of zinc in the femurs. The results suggest that the regional diet of Manaus needs to be supplemented with vitamin A to maintain the

  5. Assessment of engineering plant analyzer with Peach Bottom 2 stability tests

    International Nuclear Information System (INIS)

    Rohatgi, U.S.; Mallen, A.N.; Cheng, H.S.; Wulff, W.

    1992-01-01

    Engineering Plant Analyzer (EPA) has been developed to simulate plant transients for Boiling Water Reactor (BWR). Recently, this code has been used to simulate LaSalle-2 instability event which was initiated by a failure in the feed water heater. The simulation was performed for the scram conditions and for the postulated failure in the scram. In order to assess the capability of the EPA to simulate oscillatory flows as observed in the LaSalle event, EPA has been benchmarked with the available data from the Peach Bottom 2 (PB2) Instability tests PT1, PT2, and PT4. This document provides a description of these tests

  6. Sensory profile of eleven peach cultivars Perfil sensorial de onze cultivares de pêssegos

    Directory of Open Access Journals (Sweden)

    Francine Lorena Cuquel

    2012-03-01

    Full Text Available The goal of this study was to evaluate the sensory profile of eleven peach cultivars grown in an experimental orchard located in the city of Lapa (PR, Brazil in two seasons. The peach cultivars analyzed were Aurora I, Chimarrita, Chiripá, Coral, Eldorado, Granada, Leonense, Maciel, Marli, Premier, and Vanguarda. The sensory analysis was performed by previously trained panelists; 20 of them in the first season and 10 in the second season. The sensory evaluation was performed using Quantitative Descriptive Analysis, in which the following attributes were measured: appearance, aroma, flesh color, flesh firmness, flavor, and juiciness. The results showed preference for sweet, soft, and juicy fruits. Chimarrita, Chiripá, and Coral fruits showed better sensorial performance than the other peach cultivars. It was also verified that the analysis of the attributes aroma, flesh firmness, and flavor is enough for performing the sensory profile of peach fruits for in natura consumption.Este trabalho teve como objetivo avaliar o perfil sensorial de onze cultivares de pêssego produzidos em duas safras em um pomar experimental implantado na Lapa (PR, Brasil. Os cultivares analisados foram Aurora I, Chimarrita, Chiripá, Coral, Eldorado, Granada, Leonense, Maciel, Marli, Premier e Vanguarda. As análises sensoriais foram realizadas por julgadores previamente treinados, sendo 20 julgadores na primeira safra e 10 na segunda. O método de avaliação empregado foi a Análise Descritiva Quantitativa na qual foram mensurados os atributos aparência, aroma, cor de polpa, firmeza de polpa, sabor e suculência dos frutos. Os resultados obtidos demonstraram a preferência por frutos de sabor adocicado, com polpa macia e suculenta. Os cultivares Chimarrita, Chiripá e Coral obtiveram o melhor desempenho nas análises sensoriais. Foi verificado ainda que os atributos aroma, firmeza de polpa e sabor são considerados suficientes para a avaliação do perfil sensorial de

  7. Endogenous hormones response to cytokinins with regard to organogenesis in explants of peach (Prunus persica L. Batsch) cultivars and rootstocks (P. persica × Prunus dulcis).

    Science.gov (United States)

    Pérez-Jiménez, Margarita; Cantero-Navarro, Elena; Pérez-Alfocea, Francisco; Cos-Terrer, José

    2014-11-01

    Organogenesis in peach (Prunus persica L. Batsch) and peach rootstocks (P. persica × Prunus dulcis) has been achieved and the action of the regeneration medium on 7 phytohormones, zeatin (Z), zeatin riboside (ZR), indole-3-acetic acid (IAA), abscisic acid (ABA), ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC), salicylic acid (SA), and jasmonic acid (JA), has been studied using High performance liquid chromatography - mass spectrometry (HPLC-MS/MS). Three scion peach cultivars, 'UFO-3', 'Flariba' and 'Alice Bigi', and the peach × almond rootstocks 'Garnem' and 'GF677' were cultured in two different media, Murashige and Skoog supplemented with plant growth regulators (PGRs) (regeneration medium) and without PGRs (control medium), in order to study the effects of the media and/or genotypes in the endogenous hormones content and their role in organogenesis. The highest regeneration rate was obtained with the peach × almond rootstocks and showed a lower content of Z, IAA, ABA, ACC and JA. Only Z, ZR and IAA were affected by the action of the culture media. This study shows which hormones are external PGRs-dependent and what is the weight of the genotype and hormones in peach organogenesis that provide an avenue to manipulate in vitro organogenesis in peach. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

  8. Relationship between endogenous hormonal content and somatic organogenesis in callus of peach (Prunus persica L. Batsch) cultivars and Prunus persica×Prunus dulcis rootstocks.

    Science.gov (United States)

    Pérez-Jiménez, Margarita; Cantero-Navarro, Elena; Pérez-Alfocea, Francisco; Le-Disquet, Isabel; Guivarc'h, Anne; Cos-Terrer, José

    2014-05-01

    The relationship between endogenous hormones content and the induction of somatic peach plant was studied. To induce multiple shoots from callus derived from the base of stem explants of the scion cultivars 'UFO-3', 'Flariba' and 'Alice Bigi', and the peach×almond rootstocks 'Garnem' and 'GF677', propagated plants were cultured on Murashige and Skoog salts augmented with 0.1mgL(-1) of indolebutyric acid, 1mgL(-1) of 6-benzylaminopurine and 3% sucrose. The highest regeneration rate was obtained with the peach×almond rootstocks. Endogenous levels of abscisic acid (ABA), indole-3-acetic acid (IAA), zeatin (Z), zeatin riboside (ZR), ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC), salicylic acid (SA), and jasmonic acid (JA) were analyzed in the organogenic callus. Lower levels of several hormones, namely Z, ZR, ABA, and ACC were found in the peach×almond rootstock compared to peach cultivars, while IAA and SA presented inconclusive returns. These results suggest that the difference in somatic organogenesis capacity observed in peach and peach×almond hybrids is markedly affected by the endogenous hormonal content of the studied genotypes. Copyright © 2014 Elsevier GmbH. All rights reserved.

  9. Investigation of the aroma of commercial peach (Prunus persica L. Batsch) types by Proton Transfer Reaction-Mass Spectrometry (PTR-MS) and sensory analysis.

    Science.gov (United States)

    Bianchi, Tiago; Weesepoel, Yannick; Koot, Alex; Iglesias, Ignasi; Eduardo, Iban; Gratacós-Cubarsí, Marta; Guerrero, Luis; Hortós, Maria; van Ruth, Saskia

    2017-09-01

    The aim of this study was to investigate the aroma and sensory profiles of various types of peaches (Prunus persica L. Batsch.). Forty-three commercial cultivars comprising peaches, flat peaches, nectarines, and canning peaches (pavías) were grown over two consecutive harvest years. Fruits were assessed for chemical aroma and sensory profiles. Chemical aroma profile was obtained by proton transfer reaction-mass spectrometry (PTR-MS) and spectral masses were tentatively identified with PTR-Time of Flight-MS (PTR-Tof-MS). Sensory analysis was performed at commercial maturity considering seven aroma/flavor attributes. The four types of peaches showed both distinct chemical aroma and sensory profiles. Flat peaches and canning peaches showed most distinct patterns according to discriminant analysis. The sensory data were related to the volatile compounds by partial least square regression. γ-Hexalactone, γ-octalactone, hotrienol, acetic acid and ethyl acetate correlated positively, and benzeneacetaldehyde, trimethylbenzene and acetaldehyde negatively to the intensities of aroma and ripe fruit sensory scores. Copyright © 2017 Elsevier Ltd. All rights reserved.

  10. Production and evaluation of mineral and nutrient contents, chemical composition, and sensory properties of ice creams fortified with laboratory-prepared peach fibre

    Science.gov (United States)

    Yangılar, Filiz

    2016-01-01

    Background In the coming years, a nutraceutical food may provide both physical and mental benefits that are commonly attributed to the active components of the food. Objective In this study, we determined the nutrient and mineral contents, sensory properties, and physical and chemical characteristics of ice creams manufactured using peach fibre at different concentrations (1 and 2%). Method A total of five experimental groups were formed: two types (from peach peel and pulp) of flour, two fibre concentrations (1 and 2%), and a control group without fibres. Results Flour obtained from peach pulp and peel was found to have a significant (pice cream samples, especially the rates of Ca, K, Mg, and P, which increased in the samples depending on the content of peach fibre. Sensory ratings and acceptability of ice creams decreased significantly with increasing peach peel fibre, whereas ice creams made with C (control) and B1 (ice creams made from 1% peach pulp fibre) was the highest scored by the panellists. Conclusions Peach fibre concentrates might be used as a good source of nutraceutical ingredients. PMID:27814781

  11. Root and aerial growth in early-maturing peach trees under two crop load treatments

    Energy Technology Data Exchange (ETDEWEB)

    Abrisqueta, I.; Conejero, W.; López-Martínez, L.; Vera, J.; Ruiz Sánchez, M.C.

    2017-07-01

    The objectives of the paper were to study the pattern of root growth (measured by minirhizotrons) in relation to trunk, fruit and shoot growth and the effects of crop load on tree growth and yield in peach trees. Two crop load (commercial and low) treatments were applied in a mature early-maturing peach tree orchard growing in Mediterranean conditions. Root growth dynamics were measured using minirhizotrons during one growing season. Shoot, trunk and fruit growth were also measured. At harvest, all fruits were weighed, counted and sized. Roots grew throughout the year but at lower rates during the active fruit growth phase. Root growth was asynchronous with shoot growth, while root and trunk growth rates were highest after harvest, when the canopy was big enough to allocate the photo-assimilates to organs that would ensure the following season’s yield. Shoot and fruit growth was greater in the low crop load treatment and was accompanied by a non-significant increase in root growth. High level of fruit thinning decreased the current yield but the fruits were more marketable because of their greater size.

  12. Distribution of the root system of peach palm under drip irrigation

    Directory of Open Access Journals (Sweden)

    Adriano da Silva Lopes

    2014-07-01

    Full Text Available The incorporation of technologies has resulted in increased productivity and the more rational management of peach palm, with irrigation being an important tool for certain regions. Thus, studies leading to proper crop management are extremely important, such as the estimate of the effective depth of the root system, which is indispensable for proper irrigation management. The objective of this study was to evaluate the effects of different irrigation depths, as applied by drip irrigation, on the distribution of the root system of peach palm. This experiment was conducted in Ilha Solteira, São Paulo State, Brazil, with drip irrigation, with the two systems (flow of 0.0023 m3 h-1 consisting of four irrigation treatments corresponding to 0, 50, 100 and 150% of Class ‘A’ pan evaporation. After five years, an analysis of the Bactris gasipaes root system was performed at a distance of 0.0, 0.5 and 1.0 meters from the trunk, collecting sampling at two depths (0.0 to 0.3 m and 0.3 to 0.6 m via the auger method (volumetric analysis. We concluded that the effective depth of the root system used for irrigation management should be a maximum of 0.3 meters.

  13. Microbiological analysis of peach palm in natura submitted to 60Co radiation

    International Nuclear Information System (INIS)

    Silva, Priscila V.; Araujo, Michel M.; Nunes, Thaise C.F.; Costa, Helbert S.F.; Villavicencio, Anna Lucia C.H.; Hojeije, Khalil Y.

    2009-01-01

    The palm tree (Bactris gasipaes Kunth) is a species with high potential benefits, because of the nutritional value of its fruits that could be used both in human and animals feeding and mainly for peach palm extraction. It represents a great source of dietary fiber and a moderate source of magnesium and iron. Food irradiation is a worldwide technology that aims to improve the product quality, in order to eliminate diverse microorganisms that can spoil the food. Radiation processing, in the recommended doses, causes very few chemical alterations and nutritional losses in foods, being considered insignificant and/or similar to other food treatments. The objective of this study was to evaluate the effect of irradiation on microbiological counts of mesophilic aerobic in the peach palm in natura. Samples were irradiated with 1.0 and 1.5 kGy using a 60 Co multipurpose irradiator. Radiation treatment appeared to be a useful alternative to reduce microbial contamination in the samples analyzed. (author)

  14. Microbiological analysis of peach palm in natura submitted to {sup 60}Co radiation

    Energy Technology Data Exchange (ETDEWEB)

    Silva, Priscila V.; Araujo, Michel M.; Nunes, Thaise C.F.; Costa, Helbert S.F.; Villavicencio, Anna Lucia C.H. [Instituto de Pesquisas Energeticas e Nucleares (IPEN-CNEN/SP), Sao Paulo, SP (Brazil)], e-mail: priscilavsilva@gmail.com, e-mail: villavic@ipen.br; Hojeije, Khalil Y. [Floresta Industria e Comercio Ltda., Sao Paulo, SP (Brazil)

    2009-07-01

    The palm tree (Bactris gasipaes Kunth) is a species with high potential benefits, because of the nutritional value of its fruits that could be used both in human and animals feeding and mainly for peach palm extraction. It represents a great source of dietary fiber and a moderate source of magnesium and iron. Food irradiation is a worldwide technology that aims to improve the product quality, in order to eliminate diverse microorganisms that can spoil the food. Radiation processing, in the recommended doses, causes very few chemical alterations and nutritional losses in foods, being considered insignificant and/or similar to other food treatments. The objective of this study was to evaluate the effect of irradiation on microbiological counts of mesophilic aerobic in the peach palm in natura. Samples were irradiated with 1.0 and 1.5 kGy using a {sup 60}Co multipurpose irradiator. Radiation treatment appeared to be a useful alternative to reduce microbial contamination in the samples analyzed. (author)

  15. Internal papillomatosis with intrahepatic cholangiocarcinoma and gastrointestinal adenocarcinoma in a peach-fronted conure (Aratinga aurea).

    Science.gov (United States)

    Gibbons, Paul M; Busch, Martin D; Tell, Lisa A; Graham, Jennifer E; Lowenstine, Linda J

    2002-01-01

    A 17-yr-old pet female peach-fronted conure (Aratinga aurea) was presented with the chief complaints of mild lethargy and weight loss with increased appetite. Antemortem diagnostics included complete blood count, plasma biochemistry, and radiography. Abnormal findings included elevated inflammatory parameters (hyperfibrinogenemia) and a space-occupying mass in the region of the liver. Histologic examination of a liver biopsy sample indicated bile duct hyperplasia leading to a presumptive diagnosis of hepatoxicosis. The bird initially showed moderate improvement with supportive care, but its condition declined 9 days after the liver biopsy. Supportive care was attempted a second time, but the bird did not improve and euthanasia was elected. Abnormal gross necropsy findings were confined to the liver, which contained multiple tan nodules that exuded yellowish fluid on cut section. Histopathologic examination revealed multicentric bile duct hyperplasia and cholangiocarcinoma as well as segmental papillary hyperplasia and adenocarcinoma in the proventriculus, ventriculus, and throughout the intestinal tract. This is the first report of concurrent internal papillomatosis, gastrointestinal adenocarcinoma, and cholangiocarcinoma in a peach-fronted conure.

  16. Trade-Off Between Fitness Gain and Cost Determines Profitability of a Peach Aphid Parasitoid.

    Science.gov (United States)

    Khatri, Diwas; He, Xiong Z; Wang, Qiao

    2016-08-01

    Aphidius colemani (Viereck) (Hymenoptera: Aphidiidae) is commercially produced and utilized for biological control of peach aphid Myzus persicae (Sulzer) (Hemiptera: Aphididae) on greenhouse crops in many countries. To provide knowledge for the evaluation of parasitoid-host interactions and development of effective mass rearing programs, we investigated how and why host age or size affected fitness gain in A. colemani We show that the parasitoid was significantly more likely to encounter larger hosts and that an encounter almost always triggered an attack attempt. However, the attack attempt did not proportionally translate into oviposition because larger aphids had greater ability to defend themselves and the parasitoid spent more time in handling larger aphids. The host age at parasitization had no effect on emergence rates and sex ratio of parasitoid progeny, suggesting that pupae and larvae have similar survival rate in hosts of different ages and/or the parasitoid females do not adjust sex allocation based on host size. When parasitizing mid-aged hosts, the parasitoid gained maximum fitness for their progeny in developmental period, body size, and parasitism. Taking all findings together, we suggest that parasitizing mid-aged green peach aphid nymphs is most profitable for A. colemani. © The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  17. Survival of Lactobacillus rhamnosus probiotic strains in peach jam during storage at different temperatures

    Directory of Open Access Journals (Sweden)

    Cinzia Lucia Randazzo

    2013-12-01

    Full Text Available The survival of six probiotic wild strains of Lactobacillus rhamnosus was compared with that of a type strain during 78 days of storage at 25 and 5 ºC in peach synthetic medium (PSM and commercial peach jam (PJ. Changes in viable cell counts, pH values, sugar content, and colour parameters were monitored. All strains exhibited better performances in PJ than in PSM, showing count values higher than 7 Log cfu g-1 up to 78 days of storage at 5 ºC. Almost all wild strains remained above the critical value of 6 Log cfu g-1 in samples stored at 25 ºC up to 45 days, while the Lb. rhamnosus GG type strain, used as control, was not able to survive later than 15 days. In the synthetic medium used, the strains showed better survival in the samples incubated at 25 ºC, remaining viable above the critical level up to 45 days of storage, except for the strain H12. The probiotic cultures added to jam did not significantly change the colour parameters of the product; however the metabolism of lactobacilli did cause changes in the pH and in the composition of sugars.

  18. Chitin enhances biocontrol of Rhodotorula mucilaginosa to postharvest decay of peaches.

    Science.gov (United States)

    Zhang, Hongyin; Yang, Qiya; Ge, Lingling; Zhang, Guochao; Zhang, Xiaoli; Zhang, Xiaoyun

    2016-07-01

    Biological control using microbial antagonists is a promising alternative approach to synthetic fungicides. However, effective biological control requires enhancing the consistency and efficacy of the antagonists used to control postharvest diseases. This study investigated the effect of chitin on the biocontrol efficacy of Rhodotorula mucilaginosa against blue mold and Rhizopus decay of peaches and on the protein expression profiles of R. mucilaginosa. The antagonistic activity of R. mucilaginosa harvested from the nutrient yeast dextrose broth (NYDB) with 0.5% chitin added was significantly improved compared with culture in NYDB without chitin. The R. mucilaginosa population cultured in chitin-supplement NYDB and nutrient yeast chitin borth (NYCB) harvested from peach wounds was more than that of R. mucilaginosa cultured in NYDB without chitin throughout the storage period except at 1 d. The protein expression profiles findings revealed that there were several differentially expressed proteins of R. mucilaginosa in the 0.5% chitin-supplemented NYDB and NYCB compared with that of R. mucilaginosa in NYDB. Most of these were cellular proteomes relating to the primary metabolic reactions such as glycoside hydrolases, phosphoribosyl pyrophosphate, and NADH dehydrogenases. Some proteins were also related to signal transmission and stress response. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. In vitro establishment of the hybrid rootstock ‘Garfi x Nemared’ (Garnem for peach

    Directory of Open Access Journals (Sweden)

    Limberg Guevara Salguero

    2016-04-01

    Full Text Available The interspecific hybrid between almond and peach, ‘Garfield x Nemared’ (Prunus dulcis (Mill D.A.Webb x Prunus persica (L. Batsch. has become very important as rootstocks for peach in Bolivia, but propagation by traditional methods of this hybrid has been very difficult. In the present study the aim was to in vitro establishment of this hybrid. As initial explants, nodal segments from mother plants, growing under controlled culture conditions, were used. For disinfection two concentrations of sodium hypochlorite (0.5 and 0.75% and time (10 and 12 min were tested. The greatest percentage of establishment was achieved using 0.75% NaClO for 12 min in an MS culture medium free of growth regulators. A 100% control of the phenols oxidation was achieved with the combination of mother plants growing under 50% shade, young buds, use of 150 mg l-1 citric acid at the end of the disinfection process and into the culture medium and then place the test tubes with the nodal segments one week in the dark.   Keywords: interspecific hybrid, Prunus, tissue culture

  20. Micropropagation of ornamental Prunus spp. and GF305 peach, a Prunus viral indicator.

    Science.gov (United States)

    Kalinina, Anna; Brown, Daniel C W

    2007-07-01

    A micropropagation approach was developed for nine ornamental Prunus species, P. americana, P. cistena, P. glandulosa, P. serrulata 'Kwanzan', P. laurocerasus, P. sargentii, P. tomentosa, P. triloba, P. virginiana 'Schubert', commercially important in North America, and GF305 peach, commonly used for Prunus virus indexing. The micropropagation cycle based on proliferation of vegetative tissues includes establishment of tissue culture through introduction of shoot meristems in vitro, shoot proliferation, root induction and plant acclimatization steps and can be completed in 5 months. A meristem sterilization protocol minimized bacterial and fungal contamination. Multiple shoot formation in ornamental Prunus was obtained through the use of 1 mg l(-1) 6-benzyladenine. For GF305 peach, alteration in the sugar composition, fructose instead of sucrose, and addition of 1 mg l(-1 )ferulic acid had a significant impact on the shoot proliferation rate and maintenance of long-term in vitro culture. Rooting and plant acclimatization conditions were improved using a two-step protocol with a 4-day root induction in indole-3-butiric acid (IBA)-containing media with consequent 3-week root elongation in IBA-free media. One-month incubation of rooted shoots in a vermiculite-based medium resulted in additional shoot and root growth and provided better acclimatization and plant recovery. The micropropagation approach can be used for maintenance of the clonal properties for Prunus spp. as well as a protocol to support meristem therapy against viral infection.

  1. Non-destructive grading of peaches by near-infrared spectrometry

    Science.gov (United States)

    Carlomagno, G.; Capozzo, L.; Attolico, G.; Distante, A.

    2004-12-01

    This paper describes an experimental study on non-destructive methods for sorting peaches according to their degree of ripeness. The method is based on near-infrared (NIR) transmittance spectrometry in the region between 730 and 900 nm. It estimates the ripeness in terms of internal sugar content and firmness. A station for acquiring the NIR signal has been designed and realized, carefully choosing between several options for each component. Four different stations have been realized and compared during the experimental phase. The signals acquired by the station have been pre-processed using a noise-reducing method based on a packets-wavelet transform. In addition, an outlier detection technique has been applied for identifying irregular behaviors inside each of the considered classes. Finally, a minimum distance classifier estimates the grade of each experimental data. The results obtained in classification show that this early version of the station enables the correct discrimination of peaches with a percentage of 82.5%.

  2. Triptolide disrupts the actin-based Sertoli-germ cells adherens junctions by inhibiting Rho GTPases expression

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Xiang; Zhao, Fang [Jiangsu Key Laboratory of Drug Screening, China Pharmaceutical University, Nanjing 210009 (China); Lv, Zhong-ming; Shi, Wei-qin [Jiangsu Provincial Center for Disease Control and Prevention, Nanjing (China); Zhang, Lu-yong, E-mail: lyzhang@cpu.edu.cn [Jiangsu Key Laboratory of Drug Screening, China Pharmaceutical University, Nanjing 210009 (China); Key Laboratory of Drug Quality Control and Pharmacovigilance, China Pharmaceutical University, Nanjing (China); State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 210009 (China); Yan, Ming, E-mail: brookming@cpu.edu.cn [Jiangsu Key Laboratory of Drug Screening, China Pharmaceutical University, Nanjing 210009 (China)

    2016-11-01

    Triptolide (TP), derived from the medicinal plant Triterygium wilfordii Hook. f. (TWHF), is a diterpene triepoxide with variety biological and pharmacological activities. However, TP has been restricted in clinical application due to its narrow therapeutic window especially in reproductive system. During spermatogenesis, Sertoli cell cytoskeleton plays an essential role in facilitating germ cell movement and cell-cell actin-based adherens junctions (AJ). At Sertoli cell-spermatid interface, the anchoring device is a kind of AJ, known as ectoplasmic specializations (ES). In this study, we demonstrate that β-actin, an important component of cytoskeleton, has been significantly down-regulated after TP treatment. TP can inhibit the expression of Rho GTPase such as, RhoA, RhoB, Cdc42 and Rac1. Downstream of Rho GTPase, Rho-associated protein kinase (ROCKs) gene expressions were also suppressed by TP. F-actin immunofluorescence proved that TP disrupts Sertoli cells cytoskeleton network. As a result of β-actin down-regulation, TP treatment increased expression of testin, which indicating ES has been disassembled. In summary, this report illustrates that TP induces cytoskeleton dysfunction and disrupts cell-cell adherens junctions via inhibition of Rho GTPases. - Highlights: • Triptolide induced the disruption of Sertoli-germ cell adherens junction. • Rho GTPases expression and actin dynamics have been suppressed by triptolide. • Actin-based adherens junction is a potential antifertility target of triptolide. • Rho-Rock is involved in the regulation of actin dynamics.

  3. Tandem duplications of a degenerated GTP-binding domain at the origin of GTPase receptors Toc159 and thylakoidal SRP

    International Nuclear Information System (INIS)

    Hernandez Torres, Jorge; Maldonado, Monica Alexandra Arias; Chomilier, Jacques

    2007-01-01

    The evolutionary origin of some nuclear encoded proteins that translocate proteins across the chloroplast envelope remains unknown. Therefore, sequences of GTPase proteins constituting the Arabidopsis thaliana translocon at the outer membrane of chloroplast (atToc) complexes were analyzed by means of HCA. In particular, atToc159 and related proteins (atToc132, atToc120, and atToc90) do not have proven homologues of prokaryotic or eukaryotic ancestry. We established that the three domains commonly referred to as A, G, and M originate from the GTPase G domain, tandemly repeated, and probably evolving toward an unstructured conformation in the case of the A domain. It resulted from this study a putative common ancestor for these proteins and a new domain definition, in particular the splitting of A into three domains (A1, A2, and A3), has been proposed. The family of Toc159, previously containing A. thaliana and Pisum sativum, has been extended to Medicago truncatula and Populus trichocarpa and it has been revised for Oryza sativa. They have also been compared to GTPase subunits involved in the cpSRP system. A distant homology has been revealed among Toc and cpSRP GTP-hydrolyzing proteins of A. thaliana, and repetitions of a GTPase domain were also found in cpSRP protein receptors, by means of HCA analysis

  4. Mutation spectrum in the large GTPase dynamin 2, and genotype-phenotype correlation in autosomal dominant centronuclear myopathy

    DEFF Research Database (Denmark)

    Böhm, Johann; Biancalana, Valérie; Dechene, Elizabeth T

    2012-01-01

    Centronuclear myopathy (CNM) is a genetically heterogeneous disorder associated with general skeletal muscle weakness, type I fiber predominance and atrophy, and abnormally centralized nuclei. Autosomal dominant CNM is due to mutations in the large GTPase dynamin 2 (DNM2), a mechanochemical enzym...

  5. A GTPase chimera illustrates an uncoupled nucleotide affinity and release rate, Providing insight into the activation mechanism

    DEFF Research Database (Denmark)

    Guilfoyle, Amy P.; Deshpande, Chandrika N.; Font Sadurni, Josep

    2014-01-01

    , biophysical studies on both the eukaryotic Gα proteins and the GTPase domain (NFeoB) of prokaryotic FeoB proteins have revealed conformational changes in the G5 loop that accompany nucleotide binding and release. However, it is unclear whether this conformational change in the G5 loop is a prerequisite...

  6. Triptolide disrupts the actin-based Sertoli-germ cells adherens junctions by inhibiting Rho GTPases expression

    International Nuclear Information System (INIS)

    Wang, Xiang; Zhao, Fang; Lv, Zhong-ming; Shi, Wei-qin; Zhang, Lu-yong; Yan, Ming

    2016-01-01

    Triptolide (TP), derived from the medicinal plant Triterygium wilfordii Hook. f. (TWHF), is a diterpene triepoxide with variety biological and pharmacological activities. However, TP has been restricted in clinical application due to its narrow therapeutic window especially in reproductive system. During spermatogenesis, Sertoli cell cytoskeleton plays an essential role in facilitating germ cell movement and cell-cell actin-based adherens junctions (AJ). At Sertoli cell-spermatid interface, the anchoring device is a kind of AJ, known as ectoplasmic specializations (ES). In this study, we demonstrate that β-actin, an important component of cytoskeleton, has been significantly down-regulated after TP treatment. TP can inhibit the expression of Rho GTPase such as, RhoA, RhoB, Cdc42 and Rac1. Downstream of Rho GTPase, Rho-associated protein kinase (ROCKs) gene expressions were also suppressed by TP. F-actin immunofluorescence proved that TP disrupts Sertoli cells cytoskeleton network. As a result of β-actin down-regulation, TP treatment increased expression of testin, which indicating ES has been disassembled. In summary, this report illustrates that TP induces cytoskeleton dysfunction and disrupts cell-cell adherens junctions via inhibition of Rho GTPases. - Highlights: • Triptolide induced the disruption of Sertoli-germ cell adherens junction. • Rho GTPases expression and actin dynamics have been suppressed by triptolide. • Actin-based adherens junction is a potential antifertility target of triptolide. • Rho-Rock is involved in the regulation of actin dynamics.

  7. Thermal inactivation of polyphenoloxidase and peroxidase in Jubileu clingstone peach and yeast isolated from its spoiled puree

    Directory of Open Access Journals (Sweden)

    Andréa Menezes Lopes

    2014-03-01

    Full Text Available The thermal inactivation of yeast isolated from spoiled Jubileu peach puree and that of polyphenoloxidase (PPO and peroxidase (POD in cv. Jubileu, which is widely cultivated in southern Rio Grande do Sul state, Brazil, were studied. PPO and POD were extracted using the protein powder method and submitted to partial purification by precipitation followed by dialysis. The enzymatic activity was determined measuring the increase in absorbance at 420 nm for PPO and 470 nm for POD. The yeast used in this investigation was isolated from spoiled Jubileu peach puree at 22 °Brix, with total initial microbial count of 22 × 10² UFCmL- 1. Stock cultures were maintained on potato dextrose agar (PDA slants at 4 °C and pH 5 for later use for microbial growth. In all cases, kinetic analysis of the results suggests that the thermal inactivation was well described by a first-order kinetic model, and the temperature dependence was significantly represented by the Arrhenius law. Both enzymes were affected by heat denaturation, and PPO was more thermostable. PPO was also more thermosTable than the yeast isolated from peach puree. The D60-values were 1.53 and 1.87 min for PPO and yeast isolated from spoiled Jubileu peach puree, respectively.

  8. [Evaluation of consumer's acceptance of a peach palm snack (Bactris gasipaes) and determination of its potential as a functional food].

    Science.gov (United States)

    López-Calvo, Rebeca; Pérez, Ana M; Ivankovich Guillén, Carmen; Calderón Villaplana, Sandra; Pineda Castro, Maria Lourdes

    2015-03-01

    The aim of this study was to evaluate consumers' acceptance of a peach palm snack and to determine its potential as a functional food by chemical characterization. An assessment was conducted with 100 consumers to determine the acceptance of different snack formulations and the results were subjected to cluster analysis. This analysis revealed two groups. Group 2 included people that consume snacks and peach palm frequently and showed the highest grades for the snack evaluated characteristics. All the consumers in group 2 and approximately 85% of the consumers in group 1 indicated that they would buy the product suggesting that there is a niche market for the developed peach palm snack. Also, a qualitative evaluation, using mini focus groups, of the two most widely accepted formulas of the snack (chosen according to previously described study) was performed. The sessions considered the opinion of middle class professionals and housewives. It was determined that the combination of tara gum and carboxymethylcellulose (CMC) allows a positive synergistic effect on the sensory characteristics of the snack, highlighting natural peach flavor and improving crunchiness. In a dry basis, the snack contains per 100 g: 9 ± 4 g of fat, 14.0 ± 0.3 g of dietary fiber, 15500 ± 32 µg of carotenoids and has an antioxidant capacity of 4700 ± 8 µmol TE, which demonstrates its potential as a functional food.

  9. Genome-wide analysis of the homeodomain-leucine zipper (HD-ZIP) gene family in peach (Prunus persica).

    Science.gov (United States)

    Zhang, C H; Ma, R J; Shen, Z J; Sun, X; Korir, N K; Yu, M L

    2014-04-08

    In this study, 33 homeodomain-leucine zipper (HD-ZIP) genes were identified in peach using the HD-ZIP amino acid sequences of Arabidopsis thaliana as a probe. Based on the phylogenetic analysis and the individual gene or protein characteristics, the HD-ZIP gene family in peach can be classified into 4 subfamilies, HD-ZIP I, II, III, and IV, containing 14, 7, 4, and 8 members, respectively. The most closely related peach HD-ZIP members within the same subfamilies shared very similar gene structure in terms of either intron/exon numbers or lengths. Almost all members of the same subfamily shared common motif compositions, thereby implying that the HD-ZIP proteins within the same subfamily may have functional similarity. The 33 peach HD-ZIP genes were distributed across scaffolds 1 to 7. Although the primary structure varied among HD-ZIP family proteins, their tertiary structures were similar. The results from this study will be useful in selecting candidate genes from specific subfamilies for functional analysis.

  10. 76 FR 31888 - Nectarines and Fresh Peaches Grown in California; Termination of Marketing Order 916 and the...

    Science.gov (United States)

    2011-06-02

    ... orders also authorize production research and marketing research and development projects, as well as the... DEPARTMENT OF AGRICULTURE Agricultural Marketing Service 7 CFR Parts 916 and 917 [Doc. No. AMS-FV... Marketing Order 916 and the Peach Provisions of Marketing Order 917 AGENCY: Agricultural Marketing Service...

  11. Effects of exogenous salicylic acid on physiological traits and CBF gene expression in peach floral organs under freezing stress

    Directory of Open Access Journals (Sweden)

    Zhang Binbin

    2017-01-01

    Full Text Available To elucidate the effects of exogenous salicylic acid (SA treatment on the cold resistance of peach flower, the floral organs of two peach cultivars were treated with 20 mg/L SA and stored at 0°C for observation and sample collection. Water application was the control. After a treatment period, the anther relative water content of the control and SA-treated flowers decreased. The extent of the reduction was greater in the control, suggesting that the SA treatment significantly helped to maintain the anther water content of peach. Analysis of the stigma relative electric conductivity revealed that the SA treatment prevented membrane injury during the low temperature treatment. Additionally, we measured CBF gene expression at low temperature in the petal, stigma and ovary. The expression was markedly upregulated in the cold-treated floral organs. CBF gene expression after SA treatment was higher than in the control when cold conditions continued. These results suggest that the effects of SA on ameliorating the freezing injury to peach floral organs and on enhancing cold tolerance may be associated with the induction of CBF gene.

  12. Differential transcript abundance and genotypic variation of four putative allergen-encoding gene families in melting peach

    NARCIS (Netherlands)

    Yang, Z.; Ma, Y.; Chen, L.; Xie, R.; Zhang, X.; Zhang, B.; Lu, M.; Wu, S.; Gilissen, L.J.W.J.; Ree, van R.; Gao, Z.

    2011-01-01

    We analysed the temporal and spatial transcript expression of the panel of 18 putative isoallergens from four gene families (Pru p 1–4) in the peach fruit, anther and leaf of two melting cultivars, to gain insight into their expression profiles and to identify the key family members. Genotypic

  13. Estimation of glycaemic index of peach palm (Bactris gasipaes) cooked fruits and chips, and pitahaya (Hylocereus spp.) pulp.

    Science.gov (United States)

    Jiménez, Gin; Gómez, Georgina; Pérez, Ana M; Blanco-Metzler, Adriana

    2012-09-01

    The glycaemic index (GI) is a physiological measure of a food's potential to increase postprandial blood glucose, as compared to the effect produced by food taken as reference, such as glucose or white bread. Currently researchers and consumers are interested in low GI foods, since their consumption is associated with better weight control and reduced risk of incidence of chronic diseases, like diabetes. In the present study, the GI value for peach palm cooked fruit, peach palm chips and pitahaya pulp was estimated. The methodology established by the FAO/WHO for determining the GI of food was used. A total of 12 healthy, non-smoking volunteers were selected and they ingested the fore mentioned foods on different occasions, in 25 g portions of available carbohydrates, after 12-14 h overnight fast. Blood glucose levels were measured in 30 min intervals up to 120 min after ingestion. Average GI value was 48 +/- 11 for the pitahaya pulp and 35 +/- 6 for the peach palm cooked fruit, which may be classified as low glyceamic index foods. The GI of peach palm chips was 60 +/- 7, corresponding to a food with a moderate GI. The processing for producing the chips caused an increase in the GI value when compared to the cooked fruit, probably because the stages of miIling, moulding and baking promote availability of starch during hydrolysis by the digestive enzymes.

  14. The Arf GTPase-activating protein family is exploited by Salmonella enterica serovar Typhimurium to invade nonphagocytic host cells.

    Science.gov (United States)

    Davidson, Anthony C; Humphreys, Daniel; Brooks, Andrew B E; Hume, Peter J; Koronakis, Vassilis

    2015-02-10

    To establish intracellular infections, Salmonella bacteria trigger host cell membrane ruffling and invasion by subverting cellular Arf guanine nucleotide exchange factors (GEFs) that activate Arf1 and Arf6 GTPases by promoting GTP binding. A family of cellular Arf GTPase-activating proteins (GAPs) can downregulate Arf signaling by stimulating GTP hydrolysis, but whether they do this during infection is unknown. Here, we uncovered a remarkable role for distinct Arf GAP family members in Salmonella invasion. The Arf6 GAPs ACAP1 and ADAP1 and the Arf1 GAP ASAP1 localized at Salmonella-induced ruffles, which was not the case for the plasma membrane-localized Arf6 GAPs ARAP3 and GIT1 or the Golgi-associated Arf1 GAP1. Surprisingly, we found that loss of ACAP1, ADAP1, or ASAP1 impaired Salmonella invasion, revealing that GAPs cannot be considered mere terminators of cytoskeleton remodeling. Salmonella invasion was restored in Arf GAP-depleted cells by expressing fast-cycling Arf derivatives, demonstrating that Arf GTP/GDP cycles facilitate Salmonella invasion. Consistent with this view, both constitutively active and dominant-negative Arf derivatives that cannot undergo GTP/GDP cycles inhibited invasion. Furthermore, we demonstrated that Arf GEFs and GAPs colocalize at invading Salmonella and collaborate to drive Arf1-dependent pathogen invasion. This study revealed that Salmonella bacteria exploit a remarkable interplay between Arf GEFs and GAPs to direct cycles of Arf GTPase activation and inactivation. These cycles drive Salmonella cytoskeleton remodeling and enable intracellular infections. To initiate infections, the Salmonella bacterial pathogen remodels the mammalian actin cytoskeleton and invades host cells by subverting host Arf GEFs that activate Arf1 and Arf6 GTPases. Cellular Arf GAPs deactivate Arf GTPases and negatively regulate cell processes, but whether they target Arfs during infection is unknown. Here, we uncovered an important role for the Arf GAP

  15. Purification, crystallization and X-ray diffraction analysis of human dynamin-related protein 1 GTPase-GED fusion protein

    International Nuclear Information System (INIS)

    Klinglmayr, Eva; Wenger, Julia; Mayr, Sandra; Bossy-Wetzel, Ella; Puehringer, Sandra

    2012-01-01

    The crystallization and initial diffraction analysis of human Drp1 GTPase-GED fusion protein are reported. The mechano-enzyme dynamin-related protein 1 plays an important role in mitochondrial fission and is implicated in cell physiology. Dysregulation of Drp1 is associated with abnormal mitochondrial dynamics and neuronal damage. Drp1 shares structural and functional similarities with dynamin 1 with respect to domain organization, ability to self-assemble into spiral-like oligomers and GTP-cycle-dependent membrane scission. Structural studies of human dynamin-1 have greatly improved the understanding of this prototypical member of the dynamin superfamily. However, high-resolution structural information for full-length human Drp1 covering the GTPase domain, the middle domain and the GTPase effector domain (GED) is still lacking. In order to obtain mechanistic insights into the catalytic activity, a nucleotide-free GTPase-GED fusion protein of human Drp1 was expressed, purified and crystallized. Initial X-ray diffraction experiments yielded data to 2.67 Å resolution. The hexagonal-shaped crystals belonged to space group P2 1 2 1 2, with unit-cell parameters a = 53.59, b = 151.65, c = 43.53 Å, one molecule per asymmetric unit and a solvent content of 42%. Expression of selenomethionine-labelled protein is currently in progress. Here, the expression, purification, crystallization and X-ray diffraction analysis of the Drp1 GTPase-GED fusion protein are presented, which form a basis for more detailed structural and biophysical analysis

  16. The Rab-GTPase-activating protein TBC1D1 regulates skeletal muscle glucose metabolism

    DEFF Research Database (Denmark)

    Szekeres, Ferenc; Chadt, Alexandra; Tom, Robby Z

    2012-01-01

    The Rab-GTPase-activating protein TBC1D1 has emerged as a novel candidate involved in metabolic regulation. Our aim was to determine whether TBC1D1 is involved in insulin as well as energy-sensing signals controlling skeletal muscle metabolism. TBC1D1-deficient congenic B6.SJL-Nob1.10 (Nob1.10(SJL...... be explained partly by a 50% reduction in GLUT4 protein, since proximal signaling at the level of Akt, AMPK, and acetyl-CoA carboxylase (ACC) was unaltered. Paradoxically, in vivo insulin-stimulated 2-deoxyglucose uptake was increased in EDL and tibialis anterior muscle from TBC1D1-deficient mice......)) and wild-type littermates were studied. Glucose and insulin tolerance, glucose utilization, hepatic glucose production, and tissue-specific insulin-mediated glucose uptake were determined. The effect of insulin, AICAR, or contraction on glucose transport was studied in isolated skeletal muscle. Glucose...

  17. Rac1 GTPase activates the WAVE regulatory complex through two distinct binding sites

    Science.gov (United States)

    Brautigam, Chad A; Xing, Wenmin; Yang, Sheng; Henry, Lisa; Doolittle, Lynda K; Walz, Thomas

    2017-01-01

    The Rho GTPase Rac1 activates the WAVE regulatory complex (WRC) to drive Arp2/3 complex-mediated actin polymerization, which underpins diverse cellular processes. Here we report the structure of a WRC-Rac1 complex determined by cryo-electron microscopy. Surprisingly, Rac1 is not located at the binding site on the Sra1 subunit of the WRC previously identified by mutagenesis and biochemical data. Rather, it binds to a distinct, conserved site on the opposite end of Sra1. Biophysical and biochemical data on WRC mutants confirm that Rac1 binds to both sites, with the newly identified site having higher affinity and both sites required for WRC activation. Our data reveal that the WRC is activated by simultaneous engagement of two Rac1 molecules, suggesting a mechanism by which cells may sense the density of active Rac1 at membranes to precisely control actin assembly. PMID:28949297

  18. Quantification of local morphodynamics and local GTPase activity by edge evolution tracking.

    Directory of Open Access Journals (Sweden)

    Yuki Tsukada

    2008-11-01

    Full Text Available Advances in time-lapse fluorescence microscopy have enabled us to directly observe dynamic cellular phenomena. Although the techniques themselves have promoted the understanding of dynamic cellular functions, the vast number of images acquired has generated a need for automated processing tools to extract statistical information. A problem underlying the analysis of time-lapse cell images is the lack of rigorous methods to extract morphodynamic properties. Here, we propose an algorithm called edge evolution tracking (EET to quantify the relationship between local morphological changes and local fluorescence intensities around a cell edge using time-lapse microscopy images. This algorithm enables us to trace the local edge extension and contraction by defining subdivided edges and their corresponding positions in successive frames. Thus, this algorithm enables the investigation of cross-correlations between local morphological changes and local intensity of fluorescent signals by considering the time shifts. By applying EET to fluorescence resonance energy transfer images of the Rho-family GTPases Rac1, Cdc42, and RhoA, we examined the cross-correlation between the local area difference and GTPase activity. The calculated correlations changed with time-shifts as expected, but surprisingly, the peak of the correlation coefficients appeared with a 6-8 min time shift of morphological changes and preceded the Rac1 or Cdc42 activities. Our method enables the quantification of the dynamics of local morphological change and local protein activity and statistical investigation of the relationship between them by considering time shifts in the relationship. Thus, this algorithm extends the value of time-lapse imaging data to better understand dynamics of cellular function.

  19. Carbohydrate-free peach (Prunus persica and plum (Prunus domestica juice affects fecal microbial ecology in an obese animal model.

    Directory of Open Access Journals (Sweden)

    Giuliana D Noratto

    Full Text Available BACKGROUND: Growing evidence shows the potential of nutritional interventions to treat obesity but most investigations have utilized non-digestible carbohydrates only. Peach and plum contain high amounts of polyphenols, compounds with demonstrated anti-obesity effects. The underlying process of successfully treating obesity using polyphenols may involve an alteration of the intestinal microbiota. However, this phenomenon is not well understood. METHODOLOGY/PRINCIPAL FINDINGS: Obese Zucker rats were assigned to three groups (peach, plum, and control, n = 10 each, wild-type group was named lean (n = 10. Carbohydrates in the fruit juices were eliminated using enzymatic hydrolysis. Fecal samples were obtained after 11 weeks of fruit or control juice administration. Real-time PCR and 454-pyrosequencing were used to evaluate changes in fecal microbiota. Over 1,500 different Operational Taxonomic Units at 97% similarity were detected in all rats. Several bacterial groups (e.g. Lactobacillus and members of Ruminococcacea were found to be more abundant in the peach but especially in the plum group (plum juice contained 3 times more total polyphenolics compared to peach juice. Principal coordinate analysis based on Unifrac-based unweighted distance matrices revealed a distinct separation between the microbiota of control and treatment groups. These changes in fecal microbiota occurred simultaneously with differences in fecal short-chain acids concentrations between the control and treatment groups as well as a significant decrease in body weight in the plum group. CONCLUSIONS: This study suggests that consumption of carbohydrate-free peach and plum juice has the potential to modify fecal microbial ecology in an obese animal model. The separate contribution of polyphenols and non-polyphenols compounds (vitamins and minerals to the observed changes is unknown.

  20. Determining The Factors Affecting Fruit Hardness of Different Peach Types with Meta Analysis

    Directory of Open Access Journals (Sweden)

    Hande Küçükönder

    2014-09-01

    Full Text Available The aim of this study is to determine the factor effective in determining the hardness of Caterina, Suidring, Royal Glory and Tirrenia peach types using meta analysis. In the study, the impact force (Fi and the contact time (tc were detected and the impulse values (I that are expressed as independent variable in the area under the curve were calculated in the measurements performed using the technique of a low-mass lateral impactor multiplicated with peach. Using the theory of elasticity, the independent variables were determined as Fmax (maximum impact force, contact time (tmax, Fmax/tmax, 1/tmax, 1/tmax2,5, Fmax/tmax 1.25 and Fmax2.5 parameters. The correlation coefficient values showing the relationship between these parameters and the dependent variable Magness-Taylor force (MT were calculated and were combined with meta-analysis by using the Hunter-Schmid and Fisher’s Z methods. The Cohen’s classification criterion was used in evaluating the resulting mean effect size (combined correlation value and in determining its direction. As a result of the meta-analysis, the mean effect size according to Hunter-Schmid method was found 0.436 (0.371-0.497 positively directed in 95% confidence interval, while it was found 0.468 (0.390-0.545 according to Fisher’s Z method. The effect sizes in both methods were determined “mid-level” according to the Cohen’s classification. When the significance level of the studies was analyzed with the Z test, all of the ones that taken into the meta analysis has been found statistically significant. As a result of the meta analysis in this study evaluating the relationship of peach types with the fruit hardness, the mean effect size has been found to reach “strong level”. Consequently, “maximum shock acceleration” was found to be a more effective factor comparing to the other factors in determining the the fruit hardness according to the results of meta analysis applied in both methods.

  1. Mites fluctuation population on peach tree (Prunus persica (L. Batsch and in associated plants

    Directory of Open Access Journals (Sweden)

    Carla Rosana Eichelberger

    2011-09-01

    Full Text Available Despite the importance of peach (Prunus persica (L. Batsch in Rio Grande do Sul, little is known about mites fluctuation population considered important to this crop. The objective of this study was to know the population diversity and fluctuation of mite species associated with Premier and Eldorado varieties in Roca Sales and Venâncio Aires counties, Rio Grande do Sul. The study was conducted from July 2008 to June 2009 when 15 plants were randomly chosen in each area. The plants were divided in quadrants and from each one a branch was chosen from which three leaves were removed: one collected in the apical region, another in the medium and the other in the basal region, totalizing 180 leaves/area. Five of the most abundant associated plants were collected monthly in enough amounts for the screening under the stereoscopic microscope during an hour. A total of 1,124 mites were found belonging to 14 families and 28 species. Tetranychus ludeni Zacher, 1913, Panonychus ulmi (Koch, 1836 and Mononychellus planki (McGregor, 1950 were the most abundant phytophagous mites, whereas Typhlodromalus aripo Deleon, 1967 and Phytoseiulus macropilis (Banks, 1904 the most common predatory mites. The period of one hour under stereoscopic microscope was enough to get a representative sample. In both places evaluated the ecologic indices were low, but little higherin Premier (H' 0.56; EqJ: 0.43 when compared to Eldorado (H' 0.53; EqJ 0.40. In Premier constant species were not observed and accessory only Brevipalpus phoenicis (Geijskes, 1939, T. ludeni and T. aripo. Higher abundance was observed in December and January and bigger amount in April. Already in Eldorado, T. ludeni and P. ulmi were constants. Greater abundance was observed in November and December, whereas grater richness in December and January. In both orchards were not found mites in buds. Tetranychus ludeni is the most abundant phytophagous mites with outbreak population in November, December and

  2. Inactivation of contaminated fungi and antioxidant effects of peach (Prunus persica L. Batsch cv Dangeumdo) by 0.5-2 kGy gamma irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Kyoung-Hee; Kim, Mi-Seon [Department of Food and Nutrition, Chungnam National University, Daejeon 305-764 (Korea, Republic of); Kim, Hong-Gi [Department of Applied Biology, Chungnam National University, Daejeon 305-764 (Korea, Republic of); Yook, Hong-Sun, E-mail: yhsuny@naver.co [Department of Food and Nutrition, Chungnam National University, Daejeon 305-764 (Korea, Republic of)

    2010-04-15

    The effect of gamma irradiation (0.5-2 kGy) on the physicochemical properties of peaches was investigated during a 6-day storage at 20+-3 deg. C. Gamma irradiation is able to inactivate the four pathogens, namely Botrytis cinerea, Penicillium expansum, Rhizopus stolonifer var. stolonifer and Monilinia fructicola in peaches. Hardness significantly decreased with the increment of irradiation dose level whereas soluble solid and total polyphenol contents increased with increment of irradiation dose level. 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity of the irradiated peach was higher than that of control, and its activity increased with increment of irradiation dose level. These results suggest that gamma irradiation of peaches improved antioxidant activity, but dramatically affects the hardness throughout the entire storage time.

  3. Prof. John Wood, Chief Executive Designate, Dr Gordon Walker, Directorate, Chief Executive, Prof. Ken J. Peach, Head of the Particle Physics Department, CLRC Rutherford Appleton Laboratory, United Kingdom

    CERN Multimedia

    Patrice Loïez

    2001-01-01

    L. to. r.: Dr. Ian Wilson, CLIC Deputy Study Leader, Prof. Ken J. Peach, Head of the Particle Physics Department, Prof. John Wood, Chief Executive Designate, Dr. Gordon Walker, Directorate, Chief Executive

  4. Inactivation of contaminated fungi and antioxidant effects of peach (Prunus persica L. Batsch cv Dangeumdo) by 0.5-2 kGy gamma irradiation

    International Nuclear Information System (INIS)

    Kim, Kyoung-Hee; Kim, Mi-Seon; Kim, Hong-Gi; Yook, Hong-Sun

    2010-01-01

    The effect of gamma irradiation (0.5-2 kGy) on the physicochemical properties of peaches was investigated during a 6-day storage at 20±3 deg. C. Gamma irradiation is able to inactivate the four pathogens, namely Botrytis cinerea, Penicillium expansum, Rhizopus stolonifer var. stolonifer and Monilinia fructicola in peaches. Hardness significantly decreased with the increment of irradiation dose level whereas soluble solid and total polyphenol contents increased with increment of irradiation dose level. 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity of the irradiated peach was higher than that of control, and its activity increased with increment of irradiation dose level. These results suggest that gamma irradiation of peaches improved antioxidant activity, but dramatically affects the hardness throughout the entire storage time.

  5. Vasoactive intestinal peptide-induced neurite remodeling in human neuroblastoma SH-SY5Y cells implicates the Cdc42 GTPase and is independent of Ras-ERK pathway

    International Nuclear Information System (INIS)

    Alleaume, Celine; Eychene, Alain; Harnois, Thomas; Bourmeyster, Nicolas; Constantin, Bruno; Caigneaux, Evelyne; Muller, Jean-Marc; Philippe, Michel

    2004-01-01

    Vasoactive intestinal peptide (VIP) is known to regulate proliferation or differentiation in normal and tumoral cells. SH-SY5Y is a differentiated cell subclone derived from the SK-N-SH human neuroblastoma cell line and possess all the components for an autocrine action of VIP. In the present study, we investigated the morphological changes and intracellular signaling pathways occurring upon VIP treatment of SH-SY5Y cells. VIP induced an early remodeling of cell projections: a branched neurite network spread out and prominent varicosities developed along neurites. Although activated by VIP, the Ras/ERK pathway was not required for the remodeling process. In contrast, pull-down experiments revealed a strong Cdc42 activation by VIP while expression of a dominant-negative Cdc42 prevented the VIP-induced neurite changes, suggesting an important role for this small GTPase in the process. These data provide the first evidence for a regulation of the activity of Rho family GTPases by VIP and bring new insights in the signaling pathways implicated in neurite remodeling process induced by VIP in neuroblastoma cells

  6. Postirradiation examination of Peach Bottom HTGR Driver Fuel Element E06-01

    International Nuclear Information System (INIS)

    Dyer, F.F.; Wichner, R.P.; Martin, W.J.; Fairchild, L.L.; Kedl, R.J.; de Nordwall, H.J.

    1976-04-01

    The report presented describes the postirradiation examinations of driver fuel element E06-01, which had been irradiated an equivalent of 384 full-power days in Peach Bottom, Unit 1. The fuel element is described in detail and its temperature and irradiation service history briefly outlined. Results presented include: (1) visual observations; (2) critical dimensions of fuel compacts, sleeve, and spine; (3) axial distributions of gamma-emitting nuclides plus 3 H and 90 Sr; (4) radial distributions of these nuclides in the sleeve and spine at three axial locations in the fueled regions and three locations in the upper reflector; (5) metallographic examination of samples of fuel compact material; and (6) burnup determinations via radiochemical analyses at two compact locations

  7. MELCOR simulation of long-term station blackout at Peach Bottom

    International Nuclear Information System (INIS)

    Madni, I.K.

    1990-01-01

    This paper presents the results from MELCOR (Version 1.8BC) calculations of the Long-Term Station Blackout Accident Sequence, with failure to depressurize the reactor vessel, at the Peach Bottom (BWR Mark I) plant, and presents comparisons with Source Term Code Package (STCP) calculations of the same sequence. This sequence assumes that batteries are available for six hours following loss of all power to the plant. Following battery failure, the reactor coolant system (RCS) inventory is boiled off through the relief valves by continued decay heat generation. This leads to core uncovery, heatup, clad oxidation, core degradation, relocation, and, eventually, vessel failure at high pressure. STCP has calculated the transient out to 13.5 hours after core uncovery. The results include the timing of key events, pressure and temperature response in the reactor vessel and containment, hydrogen production, and the release of source terms to the environment. 12 refs., 23 figs., 3 tabs

  8. TDZ pulsing evaluation on the in vitro morphogenesis of peach palm.

    Science.gov (United States)

    Graner, Erika Mendes; Oberschelp, Gustavo Pedro Javier; Brondani, Gilvano Ebling; Batagin-Piotto, Katherine Derlene; de Almeida, Cristina Vieira; de Almeida, Marcílio

    2013-04-01

    Peach palm (Bactris gasipaes Kunth.) cropping is an excellent alternative to native species exploitation; nevertheless, the problems with seed germination and conventional propagation justify the use of in vitro culturing. Aiming to asses TDZ pulsing effect on B. gasipaes morphogenesis, explants obtained from unarmed microplants were maintained in two treatments, half of them in MS free medium (without growth regulator) and the other half in MS with TDZ (0.36 μM). Both groups were transferred to growth regulator-free MS medium following 14 days of culture. After 84 days of culture, TDZ pulsing increased the growth and development of the shoots, restricted the growth and development of the roots, with no influence on adventitious bud induction or somatic embryogenesis. Furthermore, development of prickles, thickening of roots and chlorotic leaves were noted under TDZ pulsing. Leaf sheath histological analysis showed an epidermal origin and no vascularization of these prickles.

  9. Postirradiation examination of recycle test elements from the Peach Bottom Reactor

    International Nuclear Information System (INIS)

    Tiegs, T.N.; Long, E.L. Jr.

    1978-12-01

    The Recycle Test Elements were a series of tests of High-Temperature Gas-Cooled Reactor fuels irradiated in Core 2 of the Peach Bottom Unit 1 Reactor. They tested a wide variety of fissile and fertile fuel types of prime interest when the tests were designed. The fuel types included UO 2 , UC 2 , (2Th,U)O 2 , (4Th,U)O 2 , ThC 2 , and ThO 2 . The mixed thorium--uranium oxides and the pure thorium oxide were tested as Biso-coated particles only, while the others were tested as both Biso- and Triso-coated particles. The Biso coatings on the fissile kernels contained the fission products inadequately but on the fertile kernels they did so acceptably. The results from accelerated and real-time tests on the particle types agreed well

  10. Cooperative problem solving capabilities of Peach-fronted Conures (Eupsittula aurea)

    DEFF Research Database (Denmark)

    Torres Ortiz, Sara; Corregidor, Alejandro; Larsen, Ole Næsbye

    We investigated basic cognitive skills of individual Peach-fronted Conures (Eupsittula aurea, PFC) using ’the string-pulling test’ and their cooperative capabilities using ’the loose-string test’. First, 2-4 strings were suspended in different configurations to the underside of a perch. A food...... reward was attached to the end of one of the strings. By varying different spatial configurations of strings and rewards, different cognitive skills could be elucidated. Two females and two males were tested. All individuals solved three out of four string configurations (four straight strings, two slant...... strings, two contact-no contact strings), but all failed in a crossed strings test. Subsequently we tested the birds with a vertical pulley that required the birds to pull the string down to move the food reward upwards and within reach. This test was only solved by the males. The latter test showed...

  11. Seasonal variation of Prunus necrotic ringspot virus concentration in almond, peach, and plum cultivars

    Directory of Open Access Journals (Sweden)

    N. Salem

    2003-08-01

    Full Text Available Levels of Prunus necrotic ringspot virus (PNRSV infection in almond, peach, and plum cultivars over the course of an entire year were determined by testing different plant parts of naturally infected trees, using the double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA. The data showed that spring was the best time of year for PNRSV detection in flowers, active growing buds, and young leaves. PNRSV detection was less reliable during the summer months. Young leaves of all cultivars were the most reliable source for distinguishing between healthy and infected plants, while flowers and buds yielded high values in some cultivars but not in others. Seasonal fluctuations in virus concentration did not follow the same pattern in all cultivars. It is therefore impossible to distinguish between infected and healthy trees on the basis of one single sampling time for all cultivars.

  12. Persistence of two neem formulations on peach leaves and fruit: effect of the distribution.

    Science.gov (United States)

    Sarais, Giorgia; Angioni, Alberto; Lai, Francesco; Cabras, Paolo; Caboni, Pierluigi

    2009-03-25

    Persistence of azadirachtins (A+B) and of the other limonoids (nimbin, salannin, deacetylnimbin, and deacetylsalannin) on peach leaves and fruits was studied using a commercial formulation (form. C) compared with an experimental formulation (form. E) prepared with coformulations allowed in organic culture. Field experiments were carried out using three concentrations: 1x, 5x, and 10x the dose recommended by the manufacturer. The EU maximum residue level (MRL) in fruits and vegetables for azadirachtin A is 1 mg/kg with a preharvest interval (PHI) of 3 days. At the recommended dose, azadirachtin A residue on fruits was not detectable (LOQ stability. This is probably due to the amount of the active ingredients that diffuse into the epicuticular wax layer thus enhancing photostability of azadirachtoids.

  13. Analysis of core damage frequency: Peach Bottom, Unit 2 internal events

    International Nuclear Information System (INIS)

    Kolaczkowski, A.M.; Cramond, W.R.; Sype, T.T.; Maloney, K.J.; Wheeler, T.A.; Daniel, S.L.

    1989-08-01

    This document contains the appendices for the accident sequence analysis of internally initiated events for the Peach Bottom, Unit 2 Nuclear Power Plant. This is one of the five plant analyses conducted as part of the NUREG-1150 effort for the Nuclear Regulatory Commission. The work performed and described here is an extensive reanalysis of that published in October 1986 as NUREG/CR-4550, Volume 4. It addresses comments from numerous reviewers and significant changes to the plant systems and procedures made since the first report. The uncertainty analysis and presentation of results are also much improved, and considerable effort was expended on an improved analysis of loss of offsite power. The content and detail of this report is directed toward PRA practitioners who need to know how the work was done and the details for use in further studies. 58 refs., 58 figs., 52 tabs

  14. Radioactive cesium deposition on rice, wheat, peach tree and soil after nuclear accident in Fukushima

    International Nuclear Information System (INIS)

    Nakanishi, T.M.; Kobayashi, N.I.; Tanoi, K.

    2013-01-01

    We present how radioactive Cs was deposited on wheat, rice, peach tree and soil after nuclear accident in Fukushima. The deposition of radioactive Cs was found as spots at the surface of the leaves, branch or trunk of the trees, as well as in soil using one of the imaging method, autoradiography. The deposited radioactive Cs was not easily washed out, even with the treatment of acid solution. When the wheat was harvested 2 months after the accident, high radioactivity of Cs was found only on the leaves developed and expanded at the time of the accident. In the case of the rice grain, most of the radioactivity was found in bran and the radioactivity was drastically reduced in milled rice. Most of the radioactive Cs accumulation in rice plants was estimated from the absorption of the Cs ion dissolved in water, rather than Cs adsorbed in soil. (author)

  15. Analysis of containment venting for the Peach Bottom Atomic Power Station

    International Nuclear Information System (INIS)

    Hanson, D.J.; Wright, R.E.; Jenkins, J.P.

    1986-01-01

    The effectiveness of containment venting as a means of preventing or mitigating the consequences of severe accidents was evaluated for Peach Bottom Atomic Power Station Units 2 and 3 (BWR-4s with Mark I containments). Results from this evaluation indicate that the effectiveness of venting in preventing containment failure is highly dependent on the severe accident sequence. Containment venting can be effective for several classes of sequences, including loss-of-coolant accidents with breaks in the containment and transients with a failure of containment heat removal. However, based on draft procedures and equipment in place at the time of the evaluation, containment venting has limited potential for further reducing the risk associated with several sequences currently identified as significant contributors to risk. Means of improving the potential for risk reduction were identified, but their influence on risk was not analyzed

  16. Perfil sensorial de iogurte light, sabor pêssego Sensory profile of peach flavored light yogurt

    Directory of Open Access Journals (Sweden)

    Ligia R. R. Santana

    2006-09-01

    Full Text Available Foi determinado o perfil sensorial descritivo de três amostras de iogurte light, sabor pêssego, pela metodologia fundamentada na Análise Descritiva Quantitativa (ADQ. A equipe sensorial definiu os termos descritores, os materiais de referência para o treinamento das qualidades e quantificações de cada um dos termos e a ficha de avaliação das amostras, de forma consensual. Dez provadores foram selecionados e rigorosamente treinados para compor a equipe definitiva, utilizando-se como critérios o poder discriminativo, reprodutibilidade e consenso dos provadores entre si. Foram gerados doze termos descritores pelo método de rede. A intensidade de cada descritor foi avaliada em cada amostra por escala não estruturada de nove centímetros, com termos de intensidade ancorados em seus extremos, sendo o mínimo à esquerda e o máximo à direita. Os dados foram analisados por ANOVA, Teste de Tukey e Análise de Componentes Principais (ACP. Os resultados indicaram que as amostras comerciais apresentaram grande variação em seus perfis sensoriais. Na ACP, ficou evidenciado que a amostra A foi caracterizada principalmente pelos atributos gosto doce, sabor pêssego e firmeza dos pedaços de fruta. A amostra B foi caracterizada pelos atributos brilho, cor pêssego e cremosidade, e a amostra C foi caracterizada pela maior intensidade dos atributos gosto ácido, adstringência, textura farinácea e aroma artificial de pêssego. De acordo com o teste de aceitação, todas as amostras apresentaram boa aceitação em todos os atributos avaliados. Desta maneira, a análise do perfil sensorial da amostra C, que contém proteína de soja na sua formulação, revelou que este ingrediente não foi percebido pelos provadores, e não interferiu na sua aceitação.The descriptive terminology and sensory profile of three samples of peach flavored light yogurt were determined using methodology based on the Quantitative Descriptive Analysis (QDA. A sensory panel

  17. Effect of peach gum polysaccharides on quality changes of white shrimp.

    Science.gov (United States)

    Yao, Xing-Cun; Chang, Cheng-Fei; Wu, Sheng-Jun

    2015-01-01

    Peach gum polysaccharides (PGPs) have both antibacterial and antioxidant activities. In this study, the retardation effect of the PGPs on the quality changes of white shrimp (Penaeus vannamei) during refrigerated storage was investigated. Shrimp samples were untreated with different concentrations of the PGPs solution and then they were stored under refrigerated conditions for 10 days. During refrigerated storage, shrimp samples were taken periodically and their total viable count, pH value, total volatile basic nitrogen, and overall acceptability score were evaluated. Compared to the control, treatment of the PGPs solution effectively retarded bacterial growth and pH changes, reduced total volatile basic nitrogen, and increased overall acceptability score of white shrimp (P. vannamei) during refrigerated storage. The results indicate that treatment of PGPs could be a promising means to preserve white shrimp (P. vannamei). Copyright © 2014 Elsevier B.V. All rights reserved.

  18. Peach Bottom Cycle 2 Low Flow Stability Tests analysis using RELAP5/PARCS

    International Nuclear Information System (INIS)

    Costa, A.L.; Salah, A.B.; D'Auria, F.

    2004-01-01

    Nowadays, the coupled codes technique, which consists in incorporating threedimensional (3D) neutron modeling of the reactor core into system codes, is extensively used for simulating transients that involve core spatial asymmetric phenomena and strong feedback effects between core neutronics and reactor loop thermal-hydraulics. So, in this work, the coupled codes technique using RELAP5/3.3-PARCS is applied to simulate stability transients in a BWR (Boiling Water Reactor). Validation has been performed against Peach Bottom-2 Low-Flow Stability Tests. In these transients dynamically complex neutron kinetics coupling with thermal-hydraulics events take place in response to a core pressure perturbation. The calculated coupled code results are herein compared against the available experimental data. (author)

  19. Regularities in the 14C assimilates supply of fruit in old peach trees

    International Nuclear Information System (INIS)

    Petrov, A.; Manolov, P.

    1977-01-01

    Autoradiography and 14 C assimilates were used in trials with five-year vase pruned trees of the Dixired peach variety. The labelled assimilates as entries from 14 CO 2 dressed shoots in the skeletal parts were transposted by a narrow phloem strip and directed either towards the fruits or down to the trunk and root system. The cumulation of labelled assimilates in fruits is determined by a series of factors. The main of them was the coincidence of the fruitbearing branchlet base with the radioactive phloem strip of the carrying it skeletal part. The leaves/fruits ratio in regard to the fruitbearing branchlet was a more slightly acting factor and in all probability contributing to the going of photoassimilates in fruits only in the case of coincidence of the basal part of fruitbearing branchlet with the radioactive steam. The transport to the fruits was both basipetal and acropetal. The 14 C assimilates stream towards the root system could get fully exhausted and stopped by branchlets covered with numerous fruits and insufficient leafage disposed on the radioactive strip. On the other hand, the basipetal 14 C assimilates stream at the base of the erected skeletal parts, caused by a strong acceptable organ the root system, hampers the labelled assimilates supply even of those fruitbearing branchlets with a low leaves/fruits ratio whose basal parts coincided with the radioactive strip. To this contributed also the great difference in the thicknesses between the skeletal part and the side fruitbearing branchlets. The examined transport type explained to some extent the slighter growth of fruits in the lower part of compact peach tree crowns reg ardless of the great leafage presence in this top parts. (author)

  20. Phytosanitary irradiation of peach fruit moth (Lepidoptera: Carposinidae) in apple fruits

    Science.gov (United States)

    Zhan, Guoping; Li, Baishu; Gao, Meixu; Liu, Bo; Wang, Yuejin; Liu, Tao; Ren, Lili

    2014-10-01

    Peach fruit moth, Carposina sasakii Matsumura, is a serious pest of many pome and stone fruits and presents a quarantine problem in some export markets. It is widely distributed in pome fruit production areas in China, Japan, Korea, North Korea and the Far Eastern Federal District of Russia. In this investigation, gamma radiation dose-response tests were conducted with late eggs (5-d-old) and various larval stages, followed by large-scale confirmatory tests on the most tolerant stage in fruit, the fifth instar. The dose-response tests, with the target radiation dose of 20 (late eggs), 40, 60, 80, 100, 120, 140, and 160 Gy (late fifth instars in vitro) respectively applied to all stages, showed that the tolerance to radiation increased with increasing age and developmental stage. The fifth instar (most advanced instar in fruits) was determined to be the most tolerant stage requiring an estimated minimum absorbed dose of 208.6 Gy (95% CI: 195.0, 226.5 Gy) to prevent adult emergence at 99.9968% efficacy (95% confidence level). In the confirmatory tests, irradiation was applied to 30,850 late fifth instars in apple fruits with a target dose of 200 Gy (171.6-227.8 Gy measured), but only 4 deformed adults emerged that died 2 d afterwards without laying eggs. A dose of 228 Gy may be recommended as a phytosanitary irradiation treatment under ambient atmosphere for the control of peach fruit moth on all commodities with an efficacy of 99.9902% at 95% confidence level.

  1. Characterization biometric and morphophysiological of Peach rootstock seeds using images of their seedling vigor.

    Science.gov (United States)

    Souza, Aline Das Graças; Smiderle, Oscar Jose; Bianchi, Valmor Joao

    2018-04-26

    This study aimed to evaluate the efficiency of using the computerized imaging seed analysis system (SAS) in the biometric and morphophysiological characterization of seeds and the initial growth of seedlings from peach rootstocks. The experimental design was completely randomized with five replicates of 20 seeds. The variables analyzed were degree of seed humidity, length and width of seeds measured by SAS technology and manual measurements, mean germination time, germination percentage, radicle length and width, taproot length, length of the aerial part and taproot/aerial part ratio. The highest seed length, germination percentage (100%) and lower germination time (11.3), were obtained with the cv. Capdeboscq while, 'Tsukuba 1', 2' and 3' had intermediate seedlings length, varying from 1.55 to 1.65 cm with mean germination times between 14.5 and 18.0 days and average germination percentage of 96%. The computerized analysis of images is fast and efficient for biometric evaluations such as seed width and length, as well as initial growth of peach tree seedlings. The cvs Capdeboscq, Flordaguard and Tsukuba 2 presented greater radicle width, length and a mean taproot/aerial part ratio equal to 2, as well as higher number of adventitious roots, which indicated a strong positive correlation between radicle length, taproot length and initial seedling growth. The continuity of the research will certainly allow the development of reliable procedures for other species, besides allowing the identification of wider alternatives for the use of this system for the expansion of knowledge in the areas of physiology and evaluation of the physiological potential of seeds. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  2. Host Preference and Performance of the Yellow Peach Moth (Conogethes punctiferalis on Chestnut Cultivars.

    Directory of Open Access Journals (Sweden)

    Yanli Du

    Full Text Available Suitability of plant tissues as food for insects varies from plant to plant. In lepidopteran insects, fitness is largely dependent on the host-finding ability of the females. Existing studies have suggested that polyphagous lepidopterans preferentially select certain host plant species for oviposition. However, the mechanisms for host recognition and selection have not been fully elucidated. For the polyphagous yellow peach moth Conogethes punctiferalis, we explored the effect of chestnut cultivar on the performance and fitness and addressed the mechanisms of plant-volatile-mediated host recognition. By carrying out laboratory experiments and field investigation on four chestnut Castanea mollissima cultivars (Huaihuang, Huaijiu, Yanhong, and Shisheng, we found that C. punctiferalis females preferentially select Huaijiu for oviposition and infestation, and caterpillars fed on Huaijiu achieved slightly greater fitness than those fed on the other three chestnut cultivars, indicating that Huaijiu was a better suitable host for C. punctiferalis. Plant volatiles played important roles in host recognition by C. punctiferalis. All seven chestnut volatile compounds, α-pinene, camphene, β-thujene, β-pinene, eucalyptol, 3-carene, and nonanal, could trigger EAG responses in C. punctiferalis. The ubiquitous plant terpenoids, α-pinene, camphene and β-pinene, and their specific combination at concentrations and proportions similar to the emissions from the four chestnut cultivars, was sufficient to elicit host recognition behavior of female C. punctiferalis. Nonanal and a mixture containing nonanal, that mimicked the emission of C. punctiferalis infested chestnut fruits, caused avoidance response. The outcome demonstrates the effects of chestnut cultivars on the performance of C. punctiferalis and reveals the preference-performance relationship between C. punctiferalis adults and their offspring. The observed olfactory plasticity in the plant

  3. Development of biocontrol agents from food microbial isolates for controlling post-harvest peach brown rot caused by Monilinia fructicola.

    Science.gov (United States)

    Zhou, Ting; Schneider, Karin E; Li, Xiu-Zhen

    2008-08-15

    An unconventional strategy of screening food microbes for biocontrol activity was used to develop biocontrol agents for controlling post-harvest peach brown rot caused by Monilinia fructicola. Forty-four microbial isolates were first screened for their biocontrol activity on apple fruit. Compared with the pathogen-only check, seven of the 44 isolates reduced brown rot incidence by >50%, including four bacteria: Bacillus sp. C06, Lactobacillus sp. C03-b and Bacillus sp. T03-c, Lactobacillus sp. P02 and three yeasts: Saccharomyces delbrueckii A50, S. cerevisiae YE-5 and S. cerevisiae A41. Eight microbial isolates were selected for testing on peaches by wound co-inoculation with mixtures of individual microbial cultures and conidial suspension of M. fructicola. Only two of them showed significant biocontrol activity after five days of incubation at 22 degrees C. Bacillus sp. C06 suppressed brown rot incidence by 92% and reduced lesion diameter by 88% compared to the pathogen-only check. Bacillus sp.T03-c reduced incidence and lesion diameter by 40% and 62%, respectively. The two isolates were compared with Pseudomonas syringae MA-4, a biocontrol agent for post-harvest peach diseases, by immersing peaches in an aliquot containing individual microbial isolates and the pathogen conidia. Treatments with isolates MA-4, C06 and T03-c significantly controlled brown rot by 91, 100, and 100% respectively. However, only isolates MA-4 and C06 significantly reduced brown rot by 80% and 15%, respectively when bacterial cells alone were applied. On naturally infected peaches, both the bacterial culture and its cell-free filtrate of the isolate C06 significantly controlled peach decay resulting in 77 and 90% reduction, respectively, whereas the treatment using only the bacterial cells generally had no effect. Isolate C06 is a single colony isolate obtained from a mesophilic cheese starter, and has been identified belonging to Bacillus amyloliquefaciens. The results have clearly

  4. Effect of Nano Packaging on Storage Life and Quality Attributes of Elberta Peach

    Directory of Open Access Journals (Sweden)

    Mohammadreza Asghari

    2017-12-01

    Full Text Available Introduction: Asaclimactericfruit, peach has a highrespiration rateandvery lowshelf life. Nowadays theuse ofappropriatepostharvesttechnologiesto increase fruit postharvest lifeis necessary.Use of nanotechnologyis considered asan effective method to increase fruit postharvest life. Nanotechnology isused extensivelyinallstages of production, processing, storage, packagingand transport ofagriculturalproducts.The objective of this study was to investigate the effect of silver and silica nanocomposites, nanosilver and polypropylene containers on shelf-life and preservation of peach qualitative characteristics (Prunuspersicacv. Elberta such as titrable acidity, soluble solids, ascorbic acid, total antioxidant and total phenolics content in cold storage after the harvest. Materials and Methods: To determine the effect of nanopackaging onshelf life andqualitativecharacteristics ofElberta peaches,the experiment was conducted in a factorial based on a completely randomized design with two treatmentsand four replications.The first factor consisted of nanopackagingnanosilver, silver and silica nanocompositesandpolypropylenepackaging, and the second factor was coldstorage periods of15, 30 and 45 daysattemperaturesfrom 0 to0/5 °C and a relative humidityof 85to 95%.In general, the experiment consisted of 36 experimental units, and each includinga container with four Elberta peach fruits.Data were analyzed by using SAS software, and Microsoft Office Excel 2010 software was used to plot the graphs.Duncan's multiple range test was used to compare the means of treatments. Results and Discussion: At the end of the maintenance period, the highest firmness of peach texture was related to nanosilver containers. The main cause of fruit softening is the destruction of cell wall components, especially pectin, which is caused by certain enzymes such as polygalactronase. Whenstorage timeincreased, polypropylene containers showed a higher weight loss compared to

  5. Defect in the GTPase activating protein (GAP) function of eIF5 causes repression of GCN4 translation.

    Science.gov (United States)

    Antony A, Charles; Alone, Pankaj V

    2017-05-13

    In eukaryotes, the eIF5 protein plays an important role in translation start site selection by providing the GAP (GTPase activating protein) function. However, in yeast translation initiation fidelity defective eIF5 G31R mutant causes preferential utilization of UUG as initiation codon and is termed as Suppressor of initiation codon (Sui - ) phenotype due to its hyper GTPase activity. The eIF5 G31R mutant dominantly represses GCN4 expression and confers sensitivity to 3-Amino-1,2,4-Trizole (3AT) induced starvation. The down-regulation of the GCN4 expression (Gcn - phenotype) in the eIF5 G31R mutant was not because of leaky scanning defects; rather was due to the utilization of upUUG initiation codons at the 5' regulatory region present between uORF1 and the main GCN4 ORF. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. Mammalian knock out cells reveal prominent roles for atlastin GTPases in ER network morphology

    Energy Technology Data Exchange (ETDEWEB)

    Zhao, Guohua; Zhu, Peng-Peng; Renvoisé, Benoît; Maldonado-Báez, Lymarie; Park, Seong Hee; Blackstone, Craig, E-mail: blackstc@ninds.nih.gov

    2016-11-15

    Atlastins are large, membrane-bound GTPases that participate in the fusion of endoplasmic reticulum (ER) tubules to generate the polygonal ER network in eukaryotes. They also regulate lipid droplet size and inhibit bone morphogenetic protein (BMP) signaling, though mechanisms remain unclear. Humans have three atlastins (ATL1, ATL2, and ATL3), and ATL1 and ATL3 are mutated in autosomal dominant hereditary spastic paraplegia and hereditary sensory neuropathies. Cellular investigations of atlastin orthologs in most yeast, plants, flies and worms are facilitated by the presence of a single or predominant isoform, but loss-of-function studies in mammalian cells are complicated by multiple, broadly-expressed paralogs. We have generated mouse NIH-3T3 cells lacking all three mammalian atlastins (Atl1/2/3) using CRISPR/Cas9-mediated gene knockout (KO). ER morphology is markedly disrupted in these triple KO cells, with prominent impairment in formation of three-way ER tubule junctions. This phenotype can be rescued by expression of distant orthologs from Saccharomyces cerevisiae (Sey1p) and Arabidopsis (ROOT HAIR DEFECTIVE3) as well as any one of the three human atlastins. Minimal, if any, changes are observed in the morphology of mitochondria and the Golgi apparatus. Alterations in BMP signaling and increased sensitivity to ER stress are also noted, though effects appear more modest. Finally, atlastins appear required for the proper differentiation of NIH-3T3 cells into an adipocyte-like phenotype. These findings have important implications for the pathogenesis of hereditary spastic paraplegias and sensory neuropathies associated with atlastin mutations. - Highlights: • NIH-3T3 cells lacking all three atlastin paralogs were generated using CRISPR/Cas9. • Cells lacking all atlastin GTPases exhibit far fewer 3-way ER tubule junctions. • ER morphology defects in atlastin knockout cells are rescued by distant plant and yeast orthologs. • Atlastin knock out cells also

  7. Rac1 GTPase regulates 11β hydroxysteroid dehydrogenase type 2 and fibrotic remodeling.

    Science.gov (United States)

    Lavall, Daniel; Schuster, Pia; Jacobs, Nadine; Kazakov, Andrey; Böhm, Michael; Laufs, Ulrich

    2017-05-05

    The aim of the study was to characterize the role of Rac1 GTPase for the mineralocorticoid receptor (MR)-mediated pro-fibrotic remodeling. Transgenic mice with cardiac overexpression of constitutively active Rac1 (RacET) develop an age-dependent phenotype with atrial dilatation, fibrosis, and atrial fibrillation. Expression of MR was similar in RacET and WT mice. The expression of 11β hydroxysteroid dehydrogenase type 2 (11β-HSD2) was age-dependently up-regulated in the atria and the left ventricles of RacET mice on mRNA and protein levels. Statin treatment inhibiting Rac1 geranylgeranylation reduced 11β-HSD2 up-regulation. Samples of human left atrial myocardium showed a positive correlation between Rac1 activity and 11β-HSD2 expression ( r = 0.7169). Immunoprecipitation showed enhanced Rac1-bound 11β-HSD2 relative to Rac1 expression in RacET mice that was diminished with statin treatment. Both basal and phorbol 12-myristate 13-acetate (PMA)-induced NADPH oxidase activity were increased in RacET and correlated positively with 11β-HSD2 expression ( r = 0.788 and r = 0.843, respectively). In cultured H9c2 cardiomyocytes, Rac1 activation with l-buthionine sulfoximine increased; Rac1 inhibition with NSC23766 decreased 11β-HSD2 mRNA and protein expression. Connective tissue growth factor (CTGF) up-regulation induced by aldosterone was prevented with NSC23766. Cardiomyocyte transfection with 11β-HSD2 siRNA abolished the aldosterone-induced CTGF up-regulation. Aldosterone-stimulated MR nuclear translocation was blocked by the 11β-HSD2 inhibitor carbenoxolone. In cardiac fibroblasts, nuclear MR translocation induced by aldosterone was inhibited with NSC23766 and spironolactone. NSC23766 prevented the aldosterone-induced proliferation and migration of cardiac fibroblasts and the up-regulation of CTGF and fibronectin. In conclusion, Rac1 GTPase regulates 11β-HSD2 expression, MR activation, and MR-mediated pro-fibrotic signaling. © 2017 by The American Society for

  8. Mammalian knock out cells reveal prominent roles for atlastin GTPases in ER network morphology

    International Nuclear Information System (INIS)

    Zhao, Guohua; Zhu, Peng-Peng; Renvoisé, Benoît; Maldonado-Báez, Lymarie; Park, Seong Hee; Blackstone, Craig

    2016-01-01

    Atlastins are large, membrane-bound GTPases that participate in the fusion of endoplasmic reticulum (ER) tubules to generate the polygonal ER network in eukaryotes. They also regulate lipid droplet size and inhibit bone morphogenetic protein (BMP) signaling, though mechanisms remain unclear. Humans have three atlastins (ATL1, ATL2, and ATL3), and ATL1 and ATL3 are mutated in autosomal dominant hereditary spastic paraplegia and hereditary sensory neuropathies. Cellular investigations of atlastin orthologs in most yeast, plants, flies and worms are facilitated by the presence of a single or predominant isoform, but loss-of-function studies in mammalian cells are complicated by multiple, broadly-expressed paralogs. We have generated mouse NIH-3T3 cells lacking all three mammalian atlastins (Atl1/2/3) using CRISPR/Cas9-mediated gene knockout (KO). ER morphology is markedly disrupted in these triple KO cells, with prominent impairment in formation of three-way ER tubule junctions. This phenotype can be rescued by expression of distant orthologs from Saccharomyces cerevisiae (Sey1p) and Arabidopsis (ROOT HAIR DEFECTIVE3) as well as any one of the three human atlastins. Minimal, if any, changes are observed in the morphology of mitochondria and the Golgi apparatus. Alterations in BMP signaling and increased sensitivity to ER stress are also noted, though effects appear more modest. Finally, atlastins appear required for the proper differentiation of NIH-3T3 cells into an adipocyte-like phenotype. These findings have important implications for the pathogenesis of hereditary spastic paraplegias and sensory neuropathies associated with atlastin mutations. - Highlights: • NIH-3T3 cells lacking all three atlastin paralogs were generated using CRISPR/Cas9. • Cells lacking all atlastin GTPases exhibit far fewer 3-way ER tubule junctions. • ER morphology defects in atlastin knockout cells are rescued by distant plant and yeast orthologs. • Atlastin knock out cells also

  9. Chlamydia abortus YhbZ, a truncated Obg family GTPase, associates with the Escherichia coli large ribosomal subunit.

    Science.gov (United States)

    Polkinghorne, Adam; Vaughan, Lloyd

    2011-01-01

    The stringent stress response is vital for bacterial survival under adverse environmental conditions. Obligate intracellular Chlamydia lack key stringent response proteins, but nevertheless can interrupt the cell cycle and enter stasis or persistence upon amino acid starvation. A possible key protein retained is YhbZ, a homologue of the ObgE guanosine triphosphatase (GTPase) superfamily connecting the stringent stress response to ribosome maturation. Curiously, chlamydial YhbZ lacks the ObgE C-terminal domain thought to be essential for binding the large ribosomal subunit. We expressed recombinant Chlamydia abortus YhbZ and showed it to be a functional GTPase, with similar activity to other Obg GTPase family members. As Chlamydia are resistant to genetic manipulation, we performed heterologous expression and gradient centrifugation experiments in Escherichia coli and found that, despite the missing C-terminal domain, C. abortus YhbZ co-fractionates with the E. coli 50S large ribosomal subunit. In addition, overexpression of chlamydial YhbZ in E. coli leads to growth defects and elongation, as reported for other Obg members. YhbZ did not complement an E. coli obgE temperature-sensitive mutant, indicating the C-terminal acidic domain may have an additional role. This data supports a role for YhbZ linking the chlamydial stress response to ribosome function and cellular growth. Copyright © 2010 Elsevier Ltd. All rights reserved.

  10. An extracellular-matrix-specific GEF-GAP interaction regulates Rho GTPase crosstalk for 3D collagen migration.

    Science.gov (United States)

    Kutys, Matthew L; Yamada, Kenneth M

    2014-09-01

    Rho-family GTPases govern distinct types of cell migration on different extracellular matrix proteins in tissue culture or three-dimensional (3D) matrices. We searched for mechanisms selectively regulating 3D cell migration in different matrix environments and discovered a form of Cdc42-RhoA crosstalk governing cell migration through a specific pair of GTPase activator and inhibitor molecules. We first identified βPix, a guanine nucleotide exchange factor (GEF), as a specific regulator of migration in 3D collagen using an affinity-precipitation-based GEF screen. Knockdown of βPix specifically blocks cell migration in fibrillar collagen microenvironments, leading to hyperactive cellular protrusion accompanied by increased collagen matrix contraction. Live FRET imaging and RNAi knockdown linked this βPix knockdown phenotype to loss of polarized Cdc42 but not Rac1 activity, accompanied by enhanced, de-localized RhoA activity. Mechanistically, collagen phospho-regulates βPix, leading to its association with srGAP1, a GTPase-activating protein (GAP), needed to suppress RhoA activity. Our results reveal a matrix-specific pathway controlling migration involving a GEF-GAP interaction of βPix with srGAP1 that is critical for maintaining suppressive crosstalk between Cdc42 and RhoA during 3D collagen migration.

  11. Expression and cytoprotective activity of the small GTPase RhoB induced by the Escherichia coli cytotoxic necrotizing factor 1

    DEFF Research Database (Denmark)

    Huelsenbeck, Stefanie C; Roggenkamp, Dennis; May, Martin

    2013-01-01

    B expression, based on the inactivation of Rho/Ras proteins. In this study, we report on a long lasting expression of RhoB in cultured cells upon activation of Rho proteins by the cytotoxic necrotizing factor 1 (CNF1) from Escherichia coli. The observations of this study highlight a new pathway involving Rac1...... without any signs of cell death. In conclusion, the cytoprotective RhoB response is not only evoked by bacterial protein toxins inactivating Rho/Ras proteins but also by the Rac1-activating toxin CNF1....

  12. Single-molecule tracking of small GTPase Rac1 uncovers spatial regulation of membrane translocation and mechanism for polarized signaling

    Science.gov (United States)

    Das, Sulagna; Yin, Taofei; Yang, Qingfen; Zhang, Jingqiao; Wu, Yi I.; Yu, Ji

    2015-01-01

    Polarized Rac1 signaling is a hallmark of many cellular functions, including cell adhesion, motility, and cell division. The two steps of Rac1 activation are its translocation to the plasma membrane and the exchange of nucleotide from GDP to GTP. It is, however, unclear whether these two processes are regulated independent of each other and what their respective roles are in polarization of Rac1 signaling. We designed a single-particle tracking (SPT) method to quantitatively analyze the kinetics of Rac1 membrane translocation in living cells. We found that the rate of Rac1 translocation was significantly elevated in protrusions during cell spreading on collagen. Furthermore, combining FRET sensor imaging with SPT measurements in the same cell, the recruitment of Rac1 was found to be polarized to an extent similar to that of the nucleotide exchange process. Statistical analysis of single-molecule trajectories and optogenetic manipulation of membrane lipids revealed that Rac1 membrane translocation precedes nucleotide exchange, and is governed primarily by interactions with phospholipids, particularly PI(3,4,5)P3, instead of protein factors. Overall, the study highlights the significance of membrane translocation in spatial Rac1 signaling, which is in addition to the traditional view focusing primarily on GEF distribution and exchange reaction. PMID:25561548

  13. The small GTPase RhoA is required to maintain spinal cord neuroepithelium organization and the neural stem cell pool

    DEFF Research Database (Denmark)

    Herzog, Dominik; Loetscher, Pirmin; van Hengel, Jolanda

    2011-01-01

    ablation. We show that, in the spinal cord neuroepithelium, RhoA is essential to localize N-cadherin and ß-catenin to AJs and maintain apical-basal polarity of neural progenitor cells. Ablation of RhoA caused the loss of AJs and severe abnormalities in the organization of cells within the neuroepithelium......Dia1), does not localize to apical AJs in which it likely stabilizes intracellular adhesion by promoting local actin polymerization and microtubule organization. Furthermore, expressing a dominant-negative form of mDia1 in neural stem/progenitor cells results in a similar phenotype compared...... with that of the RhoA conditional knock-out, namely the loss of AJs and apical polarity. Together, our data show that RhoA signaling is necessary for AJ regulation and for the maintenance of mammalian neuroepithelium organization preventing precocious cell-cycle exit and differentiation....

  14. A central role for the small GTPase Rac1 in hippocampal plasticity and spatial learning and memory

    DEFF Research Database (Denmark)

    Haditsch, Ursula; Leone, Dino P; Farinelli, Mélissa

    2009-01-01

    in excitatory neurons in the forebrain in vivo not only affects spine structure, but also impairs synaptic plasticity in the hippocampus with consequent defects in hippocampus-dependent spatial learning. Furthermore, Rac1 mutants display deficits in working/episodic-like memory in the delayed matching...

  15. Gonadotropin-releasing hormone receptor activates GTPase RhoA and inhibits cell invasion in the breast cancer cell line MDA-MB-231

    International Nuclear Information System (INIS)

    Aguilar-Rojas, Arturo; Huerta-Reyes, Maira; Maya-Núñez, Guadalupe; Arechavaleta-Velásco, Fabián; Conn, P Michael; Ulloa-Aguirre, Alfredo; Valdés, Jesús

    2012-01-01

    Gonadotropin-releasing hormone (GnRH) and its receptor (GnRHR) are both expressed by a number of malignant tumors, including those of the breast. In the latter, both behave as potent inhibitors of invasion. Nevertheless, the signaling pathways whereby the activated GnRH/GnRHR system exerts this effect have not been clearly established. In this study, we provide experimental evidence that describes components of the mechanism(s) whereby GnRH inhibits breast cancer cell invasion. Actin polymerization and substrate adhesion was measured in the highly invasive cell line, MDA-MB-231 transiently expressing the wild-type or mutant DesK191 GnRHR by fluorometry, flow cytometric analysis, and confocal microscopy, in the absence or presence of GnRH agonist. The effect of RhoA-GTP on stress fiber formation and focal adhesion assembly was measured in MDA-MB-231 cells co-expressing the GnRHRs and the GAP domain of human p190Rho GAP-A or the dominant negative mutant GAP-Y1284D. Cell invasion was determined by the transwell migration assay. Agonist-stimulated activation of the wild-type GnRHR and the highly plasma membrane expressed mutant GnRHR-DesK191 transiently transfected to MDA-MB-231 cells, favored F-actin polymerization and substrate adhesion. Confocal imaging allowed detection of an association between F-actin levels and the increase in stress fibers promoted by exposure to GnRH. Pull-down assays showed that the effects observed on actin cytoskeleton resulted from GnRH-stimulated activation of RhoA GTPase. Activation of this small G protein favored the marked increase in both cell adhesion to Collagen-I and number of focal adhesion complexes leading to inhibition of the invasion capacity of MDA-MB-231 cells as disclosed by assays in Transwell Chambers. We here show that GnRH inhibits invasion of highly invasive breast cancer-derived MDA-MB-231 cells. This effect is mediated through an increase in substrate adhesion promoted by activation of RhoA GTPase and formation of

  16. Genetic differentiation and trade among populations of peach palm ( Bactris gasipaes Kunth) in the Peruvian Amazon-implications for genetic resource management.

    Science.gov (United States)

    Adin, A; Weber, J C; Sotelo Montes, C; Vidaurre, H; Vosman, B; Smulders, M J M

    2004-05-01

    Peach palm ( Bactris gasipaes Kunth) is cultivated for fruit and 'heart of palm', and is an important component of agroforestry systems in the Peruvian Amazon. In this study, AFLP was used to compare genetic diversity among domesticated populations along the Paranapura and Cuiparillo rivers, which are managed by indigenous and colonist farming communities, respectively. Gene diversity was 0.2629 for the populations in indigenous communities and 0.2534 in colonist communities. Genetic differentiation among populations ( G(st)) was 0.0377-0.0416 ( Prodents is thought to occur only across relatively short distances (100-200 m), it is likely that exchange of material by farmers and commercial traders is responsible for most of the 'long-distance' (over more than 20 km) gene flow among populations along the two rivers studied. This exchange of material may be important to counteract the effects of selection as well as genetic drift in small groups of trees in farmers' fields, much as in a metapopulation, and may account for the weak genetic differentiation between the two rivers ( G(st)=0.0249, PPeru and Brazil showed the existence of an isolation-by-distance structure up to 3,000 km, consistent with gene flow on a regional scale, likely mediated by trade in the Amazon Basin. Results are discussed with regard to practical implications for the management of genetic resources with farming communities.

  17. Effect Of Intercropping System On Green Peach Aphid Dinamics On Organic Farming Of Potato In Karo Highland

    OpenAIRE

    Lamria Sidauruk; Darma Bakti; Retna Astuti Kuswardani; Chairani Hanum

    2015-01-01

    Abstract Green peach aphid Myzus persicae Sulzer represents one of the major pest affecting decreased production which found in different potato fields in Karo Highland. This study was conducted to determine the population dynamics of Myzus persicae Sulzer on potato cropping system. The experiment was laid out in split plot design with main plot are farming system such as conventional farming semi organic farming and organic farming. The sub plot are intercropping system consist of potato mon...

  18. Classification and Discrimination of Different Fungal Diseases of Three Infection Levels on Peaches Using Hyperspectral Reflectance Imaging Analysis

    Directory of Open Access Journals (Sweden)

    Ye Sun

    2018-04-01

    Full Text Available Peaches are susceptible to infection from several postharvest diseases. In order to control disease and avoid potential health risks, it is important to identify suitable treatments for each disease type. In this study, the spectral and imaging information from hyperspectral reflectance (400~1000 nm was used to evaluate and classify three kinds of common peach disease. To reduce the large dimensionality of the hyperspectral imaging, principal component analysis (PCA was applied to analyse each wavelength image as a whole, and the first principal component was selected to extract the imaging features. A total of 54 parameters were extracted as imaging features for one sample. Three decayed stages (slight, moderate and severe decayed peaches were considered for classification by deep belief network (DBN and partial least squares discriminant analysis (PLSDA in this study. The results showed that the DBN model has better classification results than the classification accuracy of the PLSDA model. The DBN model based on integrated information (494 features showed the highest classification results for the three diseases, with accuracies of 82.5%, 92.5%, and 100% for slightly-decayed, moderately-decayed and severely-decayed samples, respectively. The successive projections algorithm (SPA was used to select the optimal features from the integrated information; then, six optimal features were selected from a total of 494 features to establish the simple model. The SPA-PLSDA model showed better results which were more feasible for industrial application. The results showed that the hyperspectral reflectance imaging technique is feasible for detecting different kinds of diseased peaches, especially at the moderately- and severely-decayed levels.

  19. Multiple R2R3-MYB transcription factors involved in the regulation of anthocyanin accumulation in peach flower

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    Hui Zhou

    2016-10-01

    Full Text Available Anthocyanin accumulation is responsible for flower coloration in peach. Here, we report the identification and functional characterization of eight flavonoid-related R2R3-MYB transcription factors, designated PpMYB10.2, PpMYB9, PpMYBPA1, Peace, PpMYB17, PpMYB18, PpMYB19 and PpMYB20, respectively, in peach flower transcriptome. PpMYB10.2 and PpMYB9 are able to activate transcription of anthocyanin biosynthetic genes, whilst PpMYBPA1 and Peace have a strong activation on the promoters of proanthocyanin (PA biosynthetic genes. PpMYB17-20 show a strong repressive effect on transcription of flavonoid pathway genes such as DFR. These results indicate that anthocyanin accumulation in peach flower is coordinately regulated by a set of R2R3-MYB genes. In addition, PpMYB9 and PpMYB10.2 are closely related but separated into two groups, designated MYB9 and MYB10, respectively. PpMYB9 shows a strong activation on the PpUGT78A2 promoter, but with no effect on the promoter of PpUGT78B (commonly called PpUFGT in previous studies. In contrast, PpMYB10.2 is able to activate the PpUFGT promoter, but not for the PpUGT78A2 promoter. Unlike the MYB10 gene that is universally present in plants, the MYB9 gene is lost in most dicot species. Therefore, the PpMYB9 gene represents a novel group of anthocyanin-related MYB activators, which may have diverged in function from the MYB10 genes. Our study will aid in understanding the complex mechanism regulating floral pigmentation in peach and functional divergence of the R2R3-MYB gene family in plants.

  20. Peach Water Relations, Gas Exchange, Growth and Shoot Mortality under Water Deficit in Semi-Arid Weather Conditions

    OpenAIRE

    Rahmati, Mitra; Davarynejad, Gholam Hossein; G?nard, Michel; Bannayan, Mohammad; Azizi, Majid; Vercambre, Gilles

    2015-01-01

    In this study the sensitivity of peach tree (Prunus persica L.) to three water stress levels from mid-pit hardening until harvest was assessed. Seasonal patterns of shoot and fruit growth, gas exchange (leaf photosynthesis, stomatal conductance and transpiration) as well as carbon (C) storage/mobilization were evaluated in relation to plant water status. A simple C balance model was also developed to investigate sink-source relationship in relation to plant water status at the tree level. The...

  1. Isolation and expression analysis of four HD-ZIP III family genes targeted by microRNA166 in peach.

    Science.gov (United States)

    Zhang, C H; Zhang, B B; Ma, R J; Yu, M L; Guo, S L; Guo, L

    2015-10-30

    MicroRNA166 (miR166) is known to have highly conserved targets that encode proteins of the class III homeodomain-leucine zipper (HD-ZIP III) family, in a broad range of plant species. To further understand the relationship between HD-ZIP III genes and miR166, four HD-ZIP III family genes (PpHB14, PpHB15, PpHB8, and PpREV) were isolated from peach (Prunus persica) tissue and characterized. Spatio-temporal expression profiles of the genes were analyzed. Genes of the peach HD-ZIP III family were predicted to encode five conserved domains. Deduced amino acid sequences and tertiary structures of the four peach HD-ZIP III genes were highly conserved, with corresponding genes in Arabidopsis thaliana. The expression level of four targets displayed the opposite trend to that of miR166 throughout fruit development, with the exception of PpHB14 from 35 to 55 days after full bloom (DAFB). This finding indicates that miR166 may negatively regulate its four targets throughout fruit development. As for leaf and phloem, the same trend in expression level was observed between four targets and miR166 from 75 to 105 DAFB. However, the opposite trend was observed for the transcript level between four targets and miR166 from 35 to 55 DAFB. miRNA166 may negatively regulate four targets in some but not all developmental stages for a given tissue. The four genes studied were observed to have, exactly or generally, the same change tendency as individual tissue development, a finding that suggests genes of the HD-ZIP III family in peach may have complementary or cooperative functions in various tissues.

  2. Incidence and phylogenetic analyses of Armillaria spp. associated with root disease in peach orchards in the State of Mexico, Mexico

    Science.gov (United States)

    R. D. Elias-Roman; R. A. Guzman-Plazola; N. B. Klopfenstein; D. Alvarado-Rosales; G. Calderon-Zavala; J. A. Mora-Aguilera; M.-S. Kim; R. Garcia-Espinosa

    2013-01-01

    Incidence of peach [Prunus persica (L.) Batsch] tree mortality attributed to Armillaria root disease was assessed from 2009 to 2011 in 15 orchards in the State of Mexico, Mexico. Incidence increased gradually every year of assessment, reaching average values of 9.7, 15.3 and 20.3% tree mortality and 23.2, 24.7 and 28.3% disease-impacted area of the orchards during 2009...

  3. Binding of the GTPase Sar1 to a Lipid Membrane Monolayer: Insertion and Orientation Studied by Infrared Reflection–Absorption Spectroscopy

    Directory of Open Access Journals (Sweden)

    Christian Schwieger

    2017-11-01

    Full Text Available Membrane-interacting proteins are polyphilic polymers that engage in dynamic protein–protein and protein–lipid interactions while undergoing changes in conformation, orientation and binding interfaces. Predicting the sites of interactions between such polypeptides and phospholipid membranes is still a challenge. One example is the small eukaryotic GTPase Sar1, which functions in phospholipid bilayer remodeling and vesicle formation as part of the multimeric coat protein complex (COPII. The membrane interaction of Sar1 is strongly dependent on its N-terminal 23 amino acids. By monolayer adsorption experiments and infrared reflection-absorption spectroscopy (IRRAS, we elucidate the role of lipids in inducing the amphipathicity of this N-terminal stretch, which inserts into the monolayer as an amphipathic helix (AH. The AH inserting angle is determined and is consistent with the philicities and spatial distribution of the amino acid monomers. Using an advanced method of IRRAS data evaluation, the orientation of Sar1 with respect to the lipid layer prior to the recruitment of further COPII proteins is determined. The result indicates that only a slight reorientation of the membrane-bound Sar1 is needed to allow coat assembly. The time-course of the IRRAS analysis corroborates a role of slow GTP hydrolysis in Sar1 desorption from the membrane.

  4. Rab5 GTPase controls chromosome alignment through Lamin disassembly and relocation of the NuMA-like protein Mud to the poles during mitosis

    Science.gov (United States)

    Capalbo, Luisa; D'Avino, Pier Paolo; Archambault, Vincent; Glover, David M.

    2011-01-01

    The small GTPase Rab5 is a conserved regulator of membrane trafficking; it regulates the formation of early endosomes, their transport along microtubules, and the fusion to the target organelles. Although several members of the endocytic pathway were recently implicated in spindle organization, it is unclear whether Rab5 has any role during mitosis. Here, we describe that Rab5 is required for proper chromosome alignment during Drosophila mitoses. We also found that Rab5 associated in vivo with nuclear Lamin and mushroom body defect (Mud), the Drosophila counterpart of nuclear mitotic apparatus protein (NuMA). Consistent with this finding, Rab5 was required for the disassembly of the nuclear envelope at mitotic entry and the accumulation of Mud at the spindle poles. Furthermore, Mud depletion caused chromosome misalignment defects that resembled the defects of Rab5 RNAi cells, and double-knockdown experiments indicated that the two proteins function in a linear pathway. Our results indicate a role for Rab5 in mitosis and reinforce the emerging view of the contributions made by cell membrane dynamics to spindle function. PMID:21987826

  5. Haploinsufficiency of a spliceosomal GTPase encoded by EFTUD2 causes mandibulofacial dysostosis with microcephaly.

    Science.gov (United States)

    Lines, Matthew A; Huang, Lijia; Schwartzentruber, Jeremy; Douglas, Stuart L; Lynch, Danielle C; Beaulieu, Chandree; Guion-Almeida, Maria Leine; Zechi-Ceide, Roseli Maria; Gener, Blanca; Gillessen-Kaesbach, Gabriele; Nava, Caroline; Baujat, Geneviève; Horn, Denise; Kini, Usha; Caliebe, Almuth; Alanay, Yasemin; Utine, Gulen Eda; Lev, Dorit; Kohlhase, Jürgen; Grix, Arthur W; Lohmann, Dietmar R; Hehr, Ute; Böhm, Detlef; Majewski, Jacek; Bulman, Dennis E; Wieczorek, Dagmar; Boycott, Kym M

    2012-02-10

    Mandibulofacial dysostosis with microcephaly (MFDM) is a rare sporadic syndrome comprising craniofacial malformations, microcephaly, developmental delay, and a recognizable dysmorphic appearance. Major sequelae, including choanal atresia, sensorineural hearing loss, and cleft palate, each occur in a significant proportion of affected individuals. We present detailed clinical findings in 12 unrelated individuals with MFDM; these 12 individuals compose the largest reported cohort to date. To define the etiology of MFDM, we employed whole-exome sequencing of four unrelated affected individuals and identified heterozygous mutations or deletions of EFTUD2 in all four. Validation studies of eight additional individuals with MFDM demonstrated causative EFTUD2 mutations in all affected individuals tested. A range of EFTUD2-mutation types, including null alleles and frameshifts, is seen in MFDM, consistent with haploinsufficiency; segregation is de novo in all cases assessed to date. U5-116kD, the protein encoded by EFTUD2, is a highly conserved spliceosomal GTPase with a central regulatory role in catalytic splicing and post-splicing-complex disassembly. MFDM is the first multiple-malformation syndrome attributed to a defect of the major spliceosome. Our findings significantly extend the range of reported spliceosomal phenotypes in humans and pave the way for further investigation in related conditions such as Treacher Collins syndrome. Copyright © 2012 The American Society of Human Genetics. Published by Elsevier Inc. All rights reserved.

  6. An overexpression screen of Toxoplasma gondii Rab-GTPases reveals distinct transport routes to the micronemes.

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    Katrin Kremer

    2013-03-01

    Full Text Available The basic organisation of the endomembrane system is conserved in all eukaryotes and comparative genome analyses provides compelling evidence that the endomembrane system of the last common eukaryotic ancestor (LCEA is complex with many genes required for regulated traffic being present. Although apicomplexan parasites, causative agents of severe human and animal diseases, appear to have only a basic set of trafficking factors such as Rab-GTPases, they evolved unique secretory organelles (micronemes, rhoptries and dense granules that are sequentially secreted during invasion of the host cell. In order to define the secretory pathway of apicomplexans, we performed an overexpression screen of Rabs in Toxoplasma gondii and identified Rab5A and Rab5C as important regulators of traffic to micronemes and rhoptries. Intriguingly, we found that not all microneme proteins traffic depends on functional Rab5A and Rab5C, indicating the existence of redundant microneme targeting pathways. Using two-colour super-resolution stimulated emission depletion (STED we verified distinct localisations of independent microneme proteins and demonstrate that micronemal organelles are organised in distinct subsets or subcompartments. Our results suggest that apicomplexan parasites modify classical regulators of the endocytic system to carryout essential parasite-specific roles in the biogenesis of their unique secretory organelles.

  7. Identification and characterization of a lymphocytic Rho-GTPase effector: rhotekin-2

    International Nuclear Information System (INIS)

    Collier, F.M.; Gregorio-King, C.C.; Gough, T.J.; Talbot, C.D.; Walder, K.; Kirkland, M.A.

    2004-01-01

    Rhotekin belongs to the group of proteins containing a Rho-binding domain that are target peptides (effectors) for the Rho-GTPases. We previously identified a novel cDNA with homology to human rhotekin and in this study we cloned and characterized the coding region of this novel 12-exon gene. The ORF encodes a 609 amino-acid protein comprising a Class I Rho-binding domain and pleckstrin homology (PH) domain. Cellular cDNA expression of this new protein, designated Rhotekin-2 (RTKN2), was shown in the cytosol and nucleus of CHO cells. Using bioinformatics and RTPCR we identified three major splice variants, which vary in both the Rho-binding and PH domains. Real-time PCR studies showed exclusive RTKN2 expression in pooled lymphocytes and further purification indicated sole expression in CD4 pos T-cells and bone marrow-derived B-cells. Gene expression was increased in quiescent T-cells but negligible in activated proliferating cells. In malignant samples expression was absent in myeloid leukaemias, low in most B-cell malignancies and CD8 pos T-cell malignancies, but very high in CD4 pos /CD8 pos T-lymphoblastic lymphoma. As the Rho family is critical in lymphocyte development and function, RTKN2 may play an important role in lymphopoiesis

  8. Maize ROP2 GTPase provides a competitive advantage to the male gametophyte.

    Science.gov (United States)

    Arthur, K M; Vejlupkova, Z; Meeley, R B; Fowler, J E

    2003-12-01

    Rop GTPases have been implicated in the regulation of plant signal transduction and cell morphogenesis. To explore ROP2 function in maize, we isolated five Mutator transposon insertions (rop2::Mu alleles). Transmission frequency through the male gametophyte, but not the female, was lower than expected in three of the rop2::Mu mutants. These three alleles formed an allelic series on the basis of the relative transmission rate of each when crossed as trans-heterozygotes. A dramatic reduction in the level of ROP2-mRNA in pollen was associated with the three alleles causing a transmission defect, whereas a rop2::Mu allele that did not result in a defect had wild-type transcript levels, thus confirming that mutation of rop2 causes the mutant phenotype. These data strongly support a role for rop2 in male gametophyte function, perhaps surprisingly, given the expression in pollen of the nearly identical duplicate gene rop9. However, the transmission defect was apparent only when a rop2::Mu heterozygote was used as the pollen donor or when a mixture of wild-type and homozygous mutant pollen was used. Thus, mutant pollen is at a competitive disadvantage compared to wild-type pollen, although mutant pollen grains lacked an obvious cellular defect. Our data demonstrate the importance in vivo of a specific Rop, rop2, in the male gametophyte.

  9. Encephalomyocarditis virus Leader protein hinge domain is responsible for interactions with Ran GTPase

    Energy Technology Data Exchange (ETDEWEB)

    Bacot-Davis, Valjean R., E-mail: bacotdavis@wisc.edu [Institute for Molecular Virology, University of Wisconsin-Madison, R.M. Bock Laboratories, 1525 Linden Dr. Madison, WI 53706 (United States); Palmenberg, Ann C., E-mail: acpalmen@wisc.edu [Institute for Molecular Virology, University of Wisconsin-Madison, R.M. Bock Laboratories, 1525 Linden Dr. Madison, WI 53706 (United States); Department of Biochemistry, University of Wisconsin-Madison, R.M. Bock Laboratories, 1525 Linden Dr. Madison, WI 53706 (United States)

    2013-08-15

    Encephalomyocarditis virus (EMCV), a Cardiovirus, initiates its polyprotein with a short 67 amino acid Leader (L) sequence. The protein acts as a unique pathogenicity factor, with anti-host activities which include the triggering of nuclear pore complex hyperphosphorylation and direct binding inhibition of the active cellular transport protein, Ran GTPase. Chemical modifications and protein mutagenesis now map the Ran binding domain to the L hinge-linker region, and in particular, to amino acids 35–40. Large deletions affecting this region were shown previously to diminish Ran binding. New point mutations, especially K35Q, D37A and W40A, preserve the intact L structure, abolish Ran binding and are deficient for nucleoporin (Nup) hyperphosphorylation. Ran itself morphs through multiple configurations, but reacts most effectively with L when in the GDP format, preferably with an empty nucleotide binding pocket. Therefore, L:Ran binding, mediated by the linker-hinge, is a required step in L-induced nuclear transport inhibition. - Highlights: • The hinge domain provides critical residues in Cardiovirus L:Ran complex formation. • Leader prefers to bind Ran in a nucleotide free, GDP-conformation. • L-induced Nup62 phosphorylation is reduced with Ran-deficient binding mutations.

  10. Sprouty regulates cell migration by inhibiting the activation of Rac1 GTPase

    International Nuclear Information System (INIS)

    Poppleton, Helen M.; Edwin, Francis; Jaggar, Laura; Ray, Ramesh; Johnson, Leonard R.; Patel, Tarun B.

    2004-01-01

    Sprouty (SPRY) protein negatively modulates fibroblast growth factor and epidermal growth factor actions. We showed that human SPRY2 inhibits cell growth and migration in response to serum and several growth factors. Using rat intestinal epithelial (IEC-6) cells, we investigated the involvement of the Rho family of GTPases, RhoA, Rac1, and cdc42 in SPRY2-mediated inhibition of cell migration and proliferation. The ability of TAT-tagged SPRY2 to inhibit proliferation and migration of IEC-6 cells transfected with constitutively active mutants of RhoA(G14V), Rac1(G12V), and cdc42 (F28L) was determined. Constitutively active RhoA(G14V), Rac1(G12V), or cdc42(F28L) did not protect cells from the anti-proliferative actions of TAT-SPRY2. The ability of TAT-hSPRY2 to inhibit migration was not altered by of RhoA(G14V) and cdc42(F28L). However, Rac1(G12V) obliterated the ability of SPRY2 to inhibit cell autonomous or serum-induced migration. Also, the activation of endogenous Rac1 was attenuated by TAT-SPRY2. Thus, SPRY2 mediates its anti-migratory actions by inhibiting Rac1 activation

  11. GTPase activity, structure, and mechanical properties of filaments assembled from bacterial cytoskeleton protein MreB.

    Science.gov (United States)

    Esue, Osigwe; Wirtz, Denis; Tseng, Yiider

    2006-02-01

    MreB, a major component of the recently discovered bacterial cytoskeleton, displays a structure homologous to its eukaryotic counterpart actin. Here, we study the assembly and mechanical properties of Thermotoga maritima MreB in the presence of different nucleotides in vitro. We found that GTP, not ADP or GDP, can mediate MreB assembly into filamentous structures as effectively as ATP. Upon MreB assembly, both GTP and ATP release the gamma phosphate at similar rates. Therefore, MreB is an equally effective ATPase and GTPase. Electron microscopy and quantitative rheology suggest that the morphologies and micromechanical properties of filamentous ATP-MreB and GTP-MreB are similar. In contrast, mammalian actin assembly is favored in the presence of ATP over GTP. These results indicate that, despite high structural homology of their monomers, T. maritima MreB and actin filaments display different assembly, morphology, micromechanics, and nucleotide-binding specificity. Furthermore, the biophysical properties of T. maritima MreB filaments, including high rigidity and propensity to form bundles, suggest a mechanism by which MreB helical structure may be involved in imposing a cylindrical architecture on rod-shaped bacterial cells.

  12. A Bacterial Pathogen Targets a Host Rab-Family GTPase Defense Pathway with a GAP.

    Science.gov (United States)

    Spanò, Stefania; Gao, Xiang; Hannemann, Sebastian; Lara-Tejero, María; Galán, Jorge E

    2016-02-10

    Cell-autonomous defense mechanisms are potent strategies that protect individual cells against intracellular pathogens. The Rab-family GTPase Rab32 was previously shown to restrict the intracellular human pathogen Salmonella Typhi, but its potential broader role in antimicrobial defense remains unknown. We show that Rab32 represents a general cell-autonomous, antimicrobial defense that is counteracted by two Salmonella effectors. Mice lacking Rab-32 or its nucleotide exchange factor BLOC-3 are permissive to S. Typhi infection and exhibit increased susceptibility to S. Typhimurium. S. Typhimurium counters this defense pathway by delivering two type III secretion effectors, SopD2, a Rab32 GAP, and GtgE, a specific Rab32 protease. An S. Typhimurium mutant strain lacking these two effectors exhibits markedly reduced virulence, which is fully restored in BLOC-3-deficient mice. These results demonstrate that a cell-autonomous, Rab32-dependent host defense pathway plays a central role in the defense against vacuolar pathogens and describe a mechanism evolved by a bacterial pathogen to counter it. Copyright © 2016 Elsevier Inc. All rights reserved.

  13. A Conserved Role for Atlastin GTPases in Regulating Lipid Droplet Size

    Directory of Open Access Journals (Sweden)

    Robin W. Klemm

    2013-05-01

    Full Text Available Lipid droplets (LDs are the major fat storage organelles in eukaryotic cells, but how their size is regulated is unknown. Using genetic screens in C. elegans for LD morphology defects in intestinal cells, we found that mutations in atlastin, a GTPase required for homotypic fusion of endoplasmic reticulum (ER membranes, cause not only ER morphology defects, but also a reduction in LD size. Similar results were obtained after depletion of atlastin or expression of a dominant-negative mutant, whereas overexpression of atlastin had the opposite effect. Atlastin depletion in Drosophila fat bodies also reduced LD size and decreased triglycerides in whole animals, sensitizing them to starvation. In mammalian cells, co-overexpression of atlastin-1 and REEP1, a paralog of the ER tubule-shaping protein DP1/REEP5, generates large LDs. The effect of atlastin-1 on LD size correlates with its activity to promote membrane fusion in vitro. Our results indicate that atlastin-mediated fusion of ER membranes is important for LD size regulation.

  14. Phosphatidylserine and GTPase activation control Cdc42 nanoclustering to counter dissipative diffusion.

    Science.gov (United States)

    Sartorel, Elodie; Ünlü, Caner; Jose, Mini; Massoni-Laporte, Aurélie; Meca, Julien; Sibarita, Jean-Baptiste; McCusker, Derek

    2018-04-18

    The anisotropic organization of plasma membrane constituents is indicative of mechanisms that drive the membrane away from equilibrium. However, defining these mechanisms is challenging due to the short spatio-temporal scales at which diffusion operates. Here, we use high-density single protein tracking combined with photoactivation localization microscopy (sptPALM) to monitor Cdc42 in budding yeast, a system in which Cdc42 exhibits anisotropic organization. Cdc42 exhibited reduced mobility at the cell pole, where it was organized in nanoclusters. The Cdc42 nanoclusters were larger at the cell pole than those observed elsewhere in the cell. These features were exacerbated in cells expressing Cdc42-GTP, and were dependent on the scaffold Bem1, which contributed to the range of mobility and nanocluster size exhibited by Cdc42. The lipid environment, in particular phosphatidylserine levels, also played a role in regulating Cdc42 nanoclustering. These studies reveal how the mobility of a Rho GTPase is controlled to counter the depletive effects of diffusion, thus stabilizing Cdc42 on the plasma membrane and sustaining cell polarity. Movie S1 Movie S1 sptPALM imaging of live yeast expressing Pil1-mEOS expressed at the genomic locus. Pil1-mEOS was simultaneously photo-converted with a 405 nm laser and imaged with a 561 nm laser using HiLo illumination. Images were acquired at 20 ms intervals, of which 300 frames are shown at 7 frames per second.

  15. The late endocytic Rab39a GTPase regulates the interaction between multivesicular bodies and chlamydial inclusions.

    Science.gov (United States)

    Gambarte Tudela, Julian; Capmany, Anahi; Romao, Maryse; Quintero, Cristian; Miserey-Lenkei, Stephanie; Raposo, Graca; Goud, Bruno; Damiani, Maria Teresa

    2015-08-15

    Given their obligate intracellular lifestyle, Chlamydia trachomatis ensure that they have access to multiple host sources of essential lipids by interfering with vesicular transport. These bacteria hijack Rab6-, Rab11- and Rab14-controlled trafficking pathways to acquire sphingomyelin from the Golgi complex. Another important source of sphingolipids, phospholipids and cholesterol are multivesicular bodies (MVBs). Despite their participation in chlamydial inclusion development and bacterial replication, the molecular mechanisms mediating the interaction between MVBs and chlamydial inclusions remain unknown. In the present study, we demonstrate that Rab39a labels a subset of late endocytic vesicles - mainly MVBs - that move along microtubules. Moreover, Rab39a is actively recruited to chlamydial inclusions throughout the pathogen life cycle by a bacterial-driven process that depends on the Rab39a GTP- or GDP-binding state. Interestingly, Rab39a participates in the delivery of MVBs and host sphingolipids to maturing chlamydial inclusions, thereby promoting inclusion growth and bacterial development. Taken together, our findings indicate that Rab39a favours chlamydial replication and infectivity. This is the first report showing that a late endocytic Rab GTPase is involved in chlamydial infection development. © 2015. Published by The Company of Biologists Ltd.

  16. Cell cycle-dependent Rho GTPase activity dynamically regulates cancer cell motility and invasion in vivo.

    Science.gov (United States)

    Kagawa, Yoshinori; Matsumoto, Shinji; Kamioka, Yuji; Mimori, Koshi; Naito, Yoko; Ishii, Taeko; Okuzaki, Daisuke; Nishida, Naohiro; Maeda, Sakae; Naito, Atsushi; Kikuta, Junichi; Nishikawa, Keizo; Nishimura, Junichi; Haraguchi, Naotsugu; Takemasa, Ichiro; Mizushima, Tsunekazu; Ikeda, Masataka; Yamamoto, Hirofumi; Sekimoto, Mitsugu; Ishii, Hideshi; Doki, Yuichiro; Matsuda, Michiyuki; Kikuchi, Akira; Mori, Masaki; Ishii, Masaru

    2013-01-01

    The mechanism behind the spatiotemporal control of cancer cell dynamics and its possible association with cell proliferation has not been well established. By exploiting the intravital imaging technique, we found that cancer cell motility and invasive properties were closely associated with the cell cycle. In vivo inoculation of human colon cancer cells bearing fluorescence ubiquitination-based cell cycle indicator (Fucci) demonstrated an unexpected phenomenon: S/G2/M cells were more motile and invasive than G1 cells. Microarray analyses showed that Arhgap11a, an uncharacterized Rho GTPase-activating protein (RhoGAP), was expressed in a cell-cycle-dependent fashion. Expression of ARHGAP11A in cancer cells suppressed RhoA-dependent mechanisms, such as stress fiber formation and focal adhesion, which made the cells more prone to migrate. We also demonstrated that RhoA suppression by ARHGAP11A induced augmentation of relative Rac1 activity, leading to an increase in the invasive properties. RNAi-based inhibition of Arhgap11a reduced the invasion and in vivo expansion of cancers. Additionally, analysis of human specimens showed the significant up-regulation of Arhgap11a in colon cancers, which was correlated with clinical invasion status. The present study suggests that ARHGAP11A, a cell cycle-dependent RhoGAP, is a critical regulator of cancer cell mobility and is thus a promising therapeutic target in invasive cancers.

  17. Cell cycle-dependent Rho GTPase activity dynamically regulates cancer cell motility and invasion in vivo.

    Directory of Open Access Journals (Sweden)

    Yoshinori Kagawa

    Full Text Available The mechanism behind the spatiotemporal control of cancer cell dynamics and its possible association with cell proliferation has not been well established. By exploiting the intravital imaging technique, we found that cancer cell motility and invasive properties were closely associated with the cell cycle. In vivo inoculation of human colon cancer cells bearing fluorescence ubiquitination-based cell cycle indicator (Fucci demonstrated an unexpected phenomenon: S/G2/M cells were more motile and invasive than G1 cells. Microarray analyses showed that Arhgap11a, an uncharacterized Rho GTPase-activating protein (RhoGAP, was expressed in a cell-cycle-dependent fashion. Expression of ARHGAP11A in cancer cells suppressed RhoA-dependent mechanisms, such as stress fiber formation and focal adhesion, which made the cells more prone to migrate. We also demonstrated that RhoA suppression by ARHGAP11A induced augmentation of relative Rac1 activity, leading to an increase in the invasive properties. RNAi-based inhibition of Arhgap11a reduced the invasion and in vivo expansion of cancers. Additionally, analysis of human specimens showed the significant up-regulation of Arhgap11a in colon cancers, which was correlated with clinical invasion status. The present study suggests that ARHGAP11A, a cell cycle-dependent RhoGAP, is a critical regulator of cancer cell mobility and is thus a promising therapeutic target in invasive cancers.

  18. The Ras GTPase-activating protein Rasal3 supports survival of naive T cells.

    Directory of Open Access Journals (Sweden)

    Ryunosuke Muro

    Full Text Available The Ras-mitogen-activated protein kinase (MAPK pathway is crucial for T cell receptor (TCR signaling in the development and function of T cells. The significance of various modulators of the Ras-MAPK pathway in T cells, however, remains to be fully understood. Ras-activating protein-like 3 (Rasal3 is an uncharacterized member of the SynGAP family that contains a conserved Ras GTPase-activating protein (GAP domain, and is predominantly expressed in the T cell lineage. In the current study, we investigated the function and physiological roles of Rasal3. Our results showed that Rasal3 possesses RasGAP activity, but not Rap1GAP activity, and represses TCR-stimulated ERK phosphorylation in a T cell line. In systemic Rasal3-deficient mice, T cell development in the thymus including positive selection, negative selection, and β-selection was unaffected. However, the number of naive, but not effector memory CD4 and CD8 T cell in the periphery was significantly reduced in Rasal3-deficient mice, and associated with a marked increase in apoptosis of these cells. Indeed, survival of Rasal3 deficient naive CD4 T cells in vivo by adoptive transfer was significantly impaired, whereas IL-7-dependent survival of naive CD4 T cells in vitro was unaltered. Collectively, Rasal3 is required for in vivo survival of peripheral naive T cells, contributing to the maintenance of optimal T cell numbers.

  19. Genome-wide identification of WRKY family genes in peach and analysis of WRKY expression during bud dormancy.

    Science.gov (United States)

    Chen, Min; Tan, Qiuping; Sun, Mingyue; Li, Dongmei; Fu, Xiling; Chen, Xiude; Xiao, Wei; Li, Ling; Gao, Dongsheng

    2016-06-01

    Bud dormancy in deciduous fruit trees is an important adaptive mechanism for their survival in cold climates. The WRKY genes participate in several developmental and physiological processes, including dormancy. However, the dormancy mechanisms of WRKY genes have not been studied in detail. We conducted a genome-wide analysis and identified 58 WRKY genes in peach. These putative genes were located on all eight chromosomes. In bioinformatics analyses, we compared the sequences of WRKY genes from peach, rice, and Arabidopsis. In a cluster analysis, the gene sequences formed three groups, of which group II was further divided into five subgroups. Gene structure was highly conserved within each group, especially in groups IId and III. Gene expression analyses by qRT-PCR showed that WRKY genes showed different expression patterns in peach buds during dormancy. The mean expression levels of six WRKY genes (Prupe.6G286000, Prupe.1G393000, Prupe.1G114800, Prupe.1G071400, Prupe.2G185100, and Prupe.2G307400) increased during endodormancy and decreased during ecodormancy, indicating that these six WRKY genes may play a role in dormancy in a perennial fruit tree. This information will be useful for selecting fruit trees with desirable dormancy characteristics or for manipulating dormancy in genetic engineering programs.

  20. Physicochemical characterization of the peach palm (Bactris gasipaes Kunth, Arecaceae fruit skin residue flour obtained by convective drying

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    Jader Martínez-Girón

    2017-09-01

    Full Text Available The production of flour from residues of peach -palm fruit skin has been a forgotten topic from the view point of its physicochemical properties, because studies reported in the literature have focused on mesocarp flour characterization. The objective of this study was to evaluate the physicochemical properties of the peach palm fruit epicarp flour obtained by convective drying at 60±2 °C through a specific production and storage process. As a result, the flour was found to be a source of bioactive compounds, showing total phenol concentration (23.40±1.30 mg gallic acid/100 g, antioxidant activity (33.10%±3.20, total carotenoids (59.31±1.61 mg β-carotene/100 g, color a* (4.95±0.58, color b* (3.25±0.57, and luminosity (33.95±3.16. On the other hand, there was a high level of total carotenoids (85% and phenolic compounds (94% retention after six months. Additionally, color change showed no significant difference during storage. In conclusion, the flour obtained is a byproduct with properties that can be used as an alternative to agroalimentary substitutes, mainly due to its color and antioxidant activity attributes. It is noteworthy to mention that this is one of the first studies to physicochemically characterize peach palm fruit skin flour from Valle del Cauca (Colombia.

  1. Elaboration of a strategy to control the peach twig borer Anarsia lineatella Zeller in the Sefrou region in Morocco

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    Asfers Adil

    2016-12-01

    Full Text Available Trapping by specific sex pheromones initiated in 2009 to monitor three pests, peach twig borer (Anarsia lineatella, oriental fruit moth (Cydia molesta and plum fruit moth (Grapholita funebrana revealed the greater importance of peach twig borer in comparison to the others. The results of monitoring the development of larval stages over time and the accumulated degree-days from biofix show that the pest develops five generations per year, one of which undergoes a diapause. In 2009 and 2010 chemical control based on tolerance threshold of 10 males/trap/2 weeks showed unsatisfactory results. With this method, the percentage of affected fruits increased from 6.8% in 2009 to 18.6% in 2010 despite the application of four treatments of organophosphate-based insecticides in 2009 and the application of four treatments in 2010 using active ingredients from different chemical families (pyrethroid, organophosphate and chlorinicotinyl. On the other hand, management of the peach twig borer by the degree-days method tested and planned on the basis of a bifenthrin treatment between 150 to 204 degree-days accumulated from biofix, gave interesting results where the percentage of affected fruits hardly exceeded 0.5% over the four years of study

  2. Screening Quality Evaluation Factors of Freeze-Dried Peach (Prunus Persica L. Batsch Powders from Different Ripening Time Cultivars

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    Chun-ju Liu

    2017-01-01

    Full Text Available The quality evaluation of processed products is complex. To simplify the quality evaluation process and improve the efficiency, fourteen evaluation factors of freeze-dried powders of seventeen cultivars of peach at different ripening times were analyzed. The most important evaluation indicators and criteria were obtained by analysis of variance (ANOVA, correlation analysis (CA, principal component analysis (PCA, system cluster analysis (SCA, and analytic hierarchy process (AHP. Results showed that the peach powders had the significant differences in quality (P<0.05, and some processing factors were related with some physicochemical and nutritional factors. Five principle components were extracted by PCA and the cumulative contribution achieved was 84.46%. Through the score plot of the first two principal components, a clear differentiation among ripening times was found and three distinct groups were separated according to ripening time. Five characteristic factors were obtained as titratable acid, browning index, hemicellulose, hygroscopicity, and vitamin C by SCA. Their weights of 0.1249, 0.3007, 0.0514, 0.4916, and 0.0315 were obtained by AHP, respectively. The peach cultivars were divided into four evaluation grades by the comprehensive quality score.

  3. Influence of microwave radiation on the post harvest decay and quality of peach fruits in cold storage

    International Nuclear Information System (INIS)

    Azarpajoun, E.; Nikkhah, SH.

    2009-01-01

    This research has been carried out in Khorasan Agricultural and Natural Resources research center to study the effect of microwave radiation on storage time and control of peach fruit rot. Peach cultivars (Alberta, Red, White and Green of Mashad) were harvested in the first and second decades of June, July and September, sorted and stored at 4°C for 12 hours. Then fruits were treated with a Microwave with the Frequency at 2450 MHZ and two intensities, low (200 w) and high (800 w) for 30, 60 and 120 seconds, the treated and control fruits were laid on in plastic trays, packed in perforated polyethylene bags and stored in cold storage (0°C and 90-95% relative humidity) for 2 months. The qualitative tests including fruit rot, total soluble solids, titrable acidity, weight loss, firmness and color were assayed after 15, 30, 45 and 60 days. Sensory attributes were measured after 60 days storage. The experimental design was factorial in frame of completely randomized design. Multiple range test (Duncan) were used to compare the means. The results showed that treating the fruit with microwave decreased the fruit rot and increased pH, flesh firmness and total soluble solids of treated peach. Microwave radiation with the 800w for 60 seconds maintained the qualitative characteristics of fruits. Panel test confirmed these results. (author)

  4. SELECTIVITY OF INSECTICIDES USED IN PEACH FARMING TO LARVAE OF Chrysoperla externa (NEUROPTERA: CHRYSOPIDAE IN SEMI-FIELD CONDITIONS

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    RODOLFO VARGAS CASTILHOS

    2017-01-01

    Full Text Available The selectivity of five insecticides, regularly used in peach farming, was assessed for larvae of the predator Chrysoperla externa (Hagen (Neuroptera: Chrysopidae by means of bioassay in semi-field conditions. The bioassay was based on the counting of captured larvae after release in peach trees treated with the insecticides (% of active ingredient in spray liquid: deltamethrin (0.001, fenthion (0.050, phosmet (0.100, lufenuron (0.005 and malathion (0.200. Bait-cards with eggs of Anagasta kuehniella (Zeller (Lepidoptera: Pyralidae were used to capture larvae from treated plants; five of them were fixed in plant canopy and five others left on the soil around stem. A protective barrier made up of galvanized steel sheet was used for each plant to avoid loss of larvae. The number of larvae feeding on the bait-cards was measured for four days. According to the number of captured larvae, each insecticide effect was estimated and classified into toxicity categories as stated by the International Organization for Biological and Integratec Control of Noxious Animals and Plants (IOBC. Based on observations, the insect growth regulator lufenuron was harmless, while the neurotoxins deltamethrin and malathion were slightly harmful; and lastly, fenthion and phosmet were moderately harmful to C. externa larvae in semi-field conditions. Thus, lufenuron should be recommended for integrated pest management, since it would preserve this predator species in peach orchards.

  5. Summary of Preliminary Criticality Analysis for Peach Bottom Fuel in the DOE Standardized Spent Nuclear Fuel Canister

    International Nuclear Information System (INIS)

    Henrikson, D.J.

    1999-01-01

    The Department of Energy's (DOE's) National Spent Nuclear Fuel Program is developing a standardized set of canisters for DOE spent nuclear fuel (SNF). These canisters will be used for DOE SNF handling, interim storage, transportation, and disposal in the national repository. Several fuels are being examined in conjunction with the DOE SNF canisters. This report summarizes the preliminary criticality safety analysis that addresses general fissile loading limits for Peach Bottom graphite fuel in the DOE SNF canister. The canister is considered both alone and inside the 5-HLW/DOE Long Spent Fuel Co-disposal Waste Package, and in intact and degraded conditions. Results are appropriate for a single DOE SNF canister. Specific facilities, equipment, canister internal structures, and scenarios for handling, storage, and transportation have not yet been defined and are not evaluated in this analysis. The analysis assumes that the DOE SNF canister is designed so that it maintains reasonable geometric integrity. Parameters important to the results are the canister outer diameter, inner diameter, and wall thickness. These parameters are assumed to have nominal dimensions of 45.7-cm (18.0-in.), 43.815-cm (17.25-in), and 0.953-cm (0.375-in.), respectively. Based on the analysis results, the recommended fissile loading for the DOE SNF canister is 13 Peach Bottom fuel elements if no internal steel is present, and 15 Peach Bottom fuel elements if credit is taken for internal steel

  6. Decontamination effects of bark washing with a high-pressure washer on Peach [Prunus persica (L.) Batsch] and Japanese Persimmon (Diospyros kaki Thunb.) contaminated with radiocaesium during dormancy

    International Nuclear Information System (INIS)

    Sato, Mamoru; Abe, Kazuhiro; Kikunaga, Hidetoshi; Ohtsuki, Tsutomu; Takata, Daisuke; Tanoi, Keitaro; Muramatsu, Yasuyuki

    2015-01-01

    The effect of bark washing with a high-pressure washer on deciduous trees contaminated during dormancy by radiocaesium fallout derived from the Fukushima Daiichi Nuclear Power Plant accident was examined using peach [Prunus persica (L.) Batsch] and Japanese persimmon (Diospyros kaki Thunb.). Eighteen-year-old peach trees ('Akatsuki') were bark-washed twice with a high-pressure washer on July 5 and 27, 2011. Seven-year-old peach trees ('Kawanakajima Hakuto') were bark-washed on January 24, 2012, and thirty-year-old Japanese persimmon trees ('Hachiya') were bark-washed on December 21, 2011. For the peach trees, most of the bark was not removed by washing with a high-pressure washer. In contrast, the rough bark of Japanese persimmon was removed completely. No significant differences in the 137 Cs concentration of 'Akatsuki' fruit were found between the treatments conducted in the summer of 2011. Upon the bark washing of peach 'Akatsuki' trees in summer, the possibility of secondary contamination of leaves via the leachate containing 137 Cs was likely. The 137 Cs concentrations in fruits and leaves of peach 'Kawanakajima Hakuto' collected in summer 2012 were decreased significantly by washing treatment conducted in winter 2011–2012. In the year after treatment, the 137 Cs concentrations in fruits and leaves of Japanese persimmon were significantly decreased by the treatment. The effect of the bark washing on decreasing 137 Cs contents in fruits and leaves was greater in Japanese persimmon than in peach. The results for 'Kawanakajima Hakuto' and 'Hachiya' demonstrated the possibility of additive contamination. (author)

  7. IQ-domain GTPase-activating protein 1 promotes the malignant phenotype of invasive ductal breast carcinoma via canonical Wnt pathway.

    Science.gov (United States)

    Zhao, Huan-Yu; Han, Yang; Wang, Jian; Yang, Lian-He; Zheng, Xiao-Ying; Du, Jiang; Wu, Guang-Ping; Wang, En-Hua

    2017-06-01

    IQ-domain GTPase-activating protein 1 is a scaffolding protein with multidomain which plays a role in modulating dishevelled (Dvl) nuclear translocation in canonical Wnt pathway. However, the biological function and mechanism of IQ-domain GTPase-activating protein 1 in invasive ductal carcinoma (IDC) remain unknown. In this study, we found that IQ-domain GTPase-activating protein 1 expression was elevated in invasive ductal carcinoma, which was positively correlated with tumor grade, lymphatic metastasis, and poor prognosis. Coexpression of IQ-domain GTPase-activating protein 1 and Dvl in the nucleus and cytoplasm of invasive ductal carcinoma was significantly correlated but not in the membrane. Postoperative survival in the patients with their coexpression in the nucleus and cytoplasm was obviously lower than that without coexpression. The positive expression rates of c-myc and cyclin D1 were significantly higher in the patients with nuclear coexpression of Dvl and IQ-domain GTPase-activating protein 1 than that with cytoplasmic coexpression, correlating with poor prognosis. IQ-domain GTPase-activating protein 1 significantly enhanced cell proliferation and invasion in invasive ductal carcinoma cell lines by interacting with Dvl in cytoplasm to promote Dvl nuclear translocation so as to upregulate the expression of c-myc and cyclin D1. Collectively, our data suggest that IQ-domain GTPase-activating protein 1 may promote the malignant phenotype of invasive ductal carcinoma via canonical Wnt signaling, and it could be used as a potential prognostic biomarker for breast cancer patients.

  8. Tolerância de frutos de pessegueiro a geadas Fruit frost tolerance in peach

    Directory of Open Access Journals (Sweden)

    André Paulo Assmann

    2008-12-01

    Full Text Available Este trabalho teve como objetivo avaliar a tolerância de frutos de pessegueiro aos danos ocasionados pela geada. A geada ocorreu no dia 5 de setembro de 2006, sendo que a temperatura mínima, a 1,5 m do solo, foi de - 1,06ºC. Foram avaliados 28 genótipos de pessegueiro em diferentes estádios fenológicos. O delineamento experimental foi o inteiramente casualizado, com três repetições (plantas de pessegueiro, procedendo-se à avaliação em seis ramos por planta. No dia em que ocorreu a geada, foram mensurados o número de frutos por ramo, diâmetro sutural médio dos frutos, número total de gemas vegetativas, percentual de brotação e percentual de brindilas formadas a partir das gemas brotadas. Uma segunda avaliação foi realizada 15 dias após a primeira, para avaliar o percentual de queda de frutos e o diâmetro sutural médio dos frutos remanescentes. Genótipos bem enfolhados e cujos frutos apresentavam endocarpo endurecido, no momento da ocorrência da geada, foram tolerantes ao dano ocasionado pelo frio; frutos com diâmetro sutural inferior a 20 mm foram suscetíveis à geada, enquanto frutos com diâmetro sutural superior a 30 mm apresentaram boa tolerância, independentemente do genótipo avaliado.The aim of this work was to evaluate the fruit damage tolerance in different peach tree genotypes after natural freeze. The frost happened on September 5, 2006, and the minimum temperature, 1.5 meters over soil, was -1.06ºC. Twenty- eight peach tree genotypes, in different stages of development, were evaluated, in a completely random design, in three replication (plants and six branches by plants. Immediately after frost, we measured the number of fruits per branches, average fruit suture diameter, percentage of sprouts and percentage of twigs in formation. A second evaluation was accomplished fifteen days after the first one in order to assess the percentage of fruit drop and the average remaining fruit suture diameter. Genotypes

  9. Contrasting patterns in the evolution of the Rab GTPase family in Archaeplastida

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    Romana Petrželková

    2014-12-01

    Full Text Available Rab GTPases are a vast group of proteins serving a role of master regulators in membrane trafficking in eukaryotes. Previous studies delineated some 23 Rab and Rab-like paralogs ancestral for eukaryotes and mapped their current phylogenetic distribution, but the analyses relied on a limited sampling of the eukaryotic diversity. Taking advantage of the recent growth of genome and transcriptome resources for phylogenetically diverse plants and algae, we reanalyzed the evolution of the Rab family in eukaryotes with the primary plastid, collectively constituting the presumably monophyletic supergroup Archaeplastida. Our most important novel findings are as follows: (i the ancestral set of Rabs in Archaeplastida included not only the paralogs Rab1, Rab2, Rab5, Rab6, Rab7, Rab8, Rab11, Rab18, Rab23, Rab24, Rab28, IFT27, and RTW (=Rabl2, as suggested previously, but also Rab14 and Rab34, because Rab14 exists in glaucophytes and Rab34 is present in glaucophytes and some green algae; (ii except in embryophytes, Rab gene duplications have been rare in Archaeplastida. Most notable is the independent emergence of divergent, possibly functionally novel, in-paralogs of Rab1 and Rab11 in several archaeplastidial lineages; (iii recurrent gene losses have been a significant factor shaping Rab gene complements in archaeplastidial species; for example, the Rab21 paralog was lost at least six times independently within Archaeplastida, once in the lineage leading to the “core” eudicots; (iv while the glaucophyte Cyanophora paradoxa has retained the highest number of ancestral Rab paralogs among all archaeplastidial species studied so far, rhodophytes underwent an extreme reduction of the Rab gene set along their stem lineage, resulting in only six paralogs (Rab1, Rab2, Rab6, Rab7, Rab11, and Rab18 present in modern red algae. Especially notable is the absence of Rab5, a virtually universal paralog essential for the endocytic pathway, suggesting that endocytosis

  10. Neuronal Rho GTPase Rac1 elimination confers neuroprotection in a mouse model of permanent ischemic stroke.

    Science.gov (United States)

    Karabiyik, Cansu; Fernandes, Rui; Figueiredo, Francisco Rosário; Socodato, Renato; Brakebusch, Cord; Lambertsen, Kate Lykke; Relvas, João Bettencourt; Santos, Sofia Duque

    2017-09-28

    The Rho GTPase Rac1 is a multifunctional protein involved in distinct pathways ranging from development to pathology. The aim of the present study was to unravel the contribution of neuronal Rac1 in regulating the response to brain injury induced by permanent focal cerebral ischemia (pMCAO). Our results show that pMCAO significantly increased total Rac1 levels in wild type mice, mainly through rising nuclear Rac1, while a reduction in Rac1 activation was observed. Such changes preceded cell death induced by excitotoxic stress. Pharmacological inhibition of Rac1 in primary neuronal cortical cells prevented the increase in oxidative stress induced after overactivation of glutamate receptors. However, this was not sufficient to prevent the associated neuronal cell death. In contrast, RNAi-mediated knock down of Rac1 in primary cortical neurons prevented cell death elicited by glutamate excitotoxicity and decreased the activity of NADPH oxidase. To test whether in vivo down regulation of neuronal Rac1 was neuroprotective after pMCAO, we used tamoxifen-inducible neuron-specific conditional Rac1-knockout mice. We observed a significant 50% decrease in brain infarct volume of knockout mice and a concomitant increase in HIF-1α expression compared to littermate control mice, demonstrating that ablation of Rac1 in neurons is neuroprotective. Transmission electron microscopy performed in the ischemic brain showed that lysosomes in the infarct of Rac1- knockout mice were preserved at similar levels to those of non-infarcted tissue, while littermate mice displayed a decrease in the number of lysosomes, further corroborating the notion that Rac1 ablation in neurons is neuroprotective. Our results demonstrate that Rac1 plays important roles in the ischemic pathological cascade and that modulation of its levels is of therapeutic interest. © 2017 International Society of Neuropathology.

  11. Functional Characterization of Monomeric GTPase Rab1 in the Secretory Pathway of Leishmania*

    Science.gov (United States)

    Bahl, Surbhi; Parashar, Smriti; Malhotra, Himanshu; Raje, Manoj; Mukhopadhyay, Amitabha

    2015-01-01

    Leishmania secretes a large number of its effectors to the extracellular milieu. However, regulation of the secretory pathway in Leishmania is not well characterized. Here, we report the cloning, expression, and characterization of the Rab1 homologue from Leishmania. We have found that LdRab1 localizes in Golgi in Leishmania. To understand the role of LdRab1 in the secretory pathway of Leishmania, we have generated transgenic parasites overexpressing GFP-LdRab1:WT, GFP-LdRab1:Q67L (a GTPase-deficient dominant positive mutant of Rab1), and GFP-LdRab1:S22N (a GDP-locked dominant negative mutant of Rab1). Surprisingly, our results have shown that overexpression of GFP-LdRab1:Q67L or GFP-LdRab1:S22N does not disrupt the trafficking and localization of hemoglobin receptor in Leishmania. To determine whether the Rab1-dependent secretory pathway is conserved in parasites, we have analyzed the role of LdRab1 in the secretion of secretory acid phosphatase and Ldgp63 in Leishmania. Our results have shown that overexpression of GFP-LdRab1:Q67L or GFP-LdRab1:S22N significantly inhibits the secretion of secretory acid phosphatase by Leishmania. We have also found that overexpression of GFP-LdRab1:Q67L or GFP-LdRab1:S22N retains RFP-Ldgp63 in Golgi and blocks the secretion of Ldgp63, whereas the trafficking of RFP-Ldgp63 in GFP-LdRab1:WT-expressing cells is unaltered in comparison with control cells. Taken together, our results have shown that the Rab1-regulated secretory pathway is well conserved, and hemoglobin receptor trafficking follows an Rab1-independent secretory pathway in Leishmania. PMID:26499792

  12. Functional Characterization of Monomeric GTPase Rab1 in the Secretory Pathway of Leishmania.

    Science.gov (United States)

    Bahl, Surbhi; Parashar, Smriti; Malhotra, Himanshu; Raje, Manoj; Mukhopadhyay, Amitabha

    2015-12-11

    Leishmania secretes a large number of its effectors to the extracellular milieu. However, regulation of the secretory pathway in Leishmania is not well characterized. Here, we report the cloning, expression, and characterization of the Rab1 homologue from Leishmania. We have found that LdRab1 localizes in Golgi in Leishmania. To understand the role of LdRab1 in the secretory pathway of Leishmania, we have generated transgenic parasites overexpressing GFP-LdRab1:WT, GFP-LdRab1:Q67L (a GTPase-deficient dominant positive mutant of Rab1), and GFP-LdRab1:S22N (a GDP-locked dominant negative mutant of Rab1). Surprisingly, our results have shown that overexpression of GFP-LdRab1:Q67L or GFP-LdRab1:S22N does not disrupt the trafficking and localization of hemoglobin receptor in Leishmania. To determine whether the Rab1-dependent secretory pathway is conserved in parasites, we have analyzed the role of LdRab1 in the secretion of secretory acid phosphatase and Ldgp63 in Leishmania. Our results have shown that overexpression of GFP-LdRab1:Q67L or GFP-LdRab1:S22N significantly inhibits the secretion of secretory acid phosphatase by Leishmania. We have also found that overexpression of GFP-LdRab1:Q67L or GFP-LdRab1:S22N retains RFP-Ldgp63 in Golgi and blocks the secretion of Ldgp63, whereas the trafficking of RFP-Ldgp63 in GFP-LdRab1:WT-expressing cells is unaltered in comparison with control cells. Taken together, our results have shown that the Rab1-regulated secretory pathway is well conserved, and hemoglobin receptor trafficking follows an Rab1-independent secretory pathway in Leishmania. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  13. Synapse Formation in Monosynaptic Sensory–Motor Connections Is Regulated by Presynaptic Rho GTPase Cdc42

    Science.gov (United States)

    Imai, Fumiyasu; Ladle, David R.; Leslie, Jennifer R.; Duan, Xin; Rizvi, Tilat A.; Ciraolo, Georgianne M.; Zheng, Yi

    2016-01-01

    Spinal reflex circuit development requires the precise regulation of axon trajectories, synaptic specificity, and synapse formation. Of these three crucial steps, the molecular mechanisms underlying synapse formation between group Ia proprioceptive sensory neurons and motor neurons is the least understood. Here, we show that the Rho GTPase Cdc42 controls synapse formation in monosynaptic sensory–motor connections in presynaptic, but not postsynaptic, neurons. In mice lacking Cdc42 in presynaptic sensory neurons, proprioceptive sensory axons appropriately reach the ventral spinal cord, but significantly fewer synapses are formed with motor neurons compared with wild-type mice. Concordantly, electrophysiological analyses show diminished EPSP amplitudes in monosynaptic sensory–motor circuits in these mutants. Temporally targeted deletion of Cdc42 in sensory neurons after sensory–motor circuit establishment reveals that Cdc42 does not affect synaptic transmission. Furthermore, addition of the synaptic organizers, neuroligins, induces presynaptic differentiation of wild-type, but not Cdc42-deficient, proprioceptive sensory neurons in vitro. Together, our findings demonstrate that Cdc42 in presynaptic neurons is required for synapse formation in monosynaptic sensory–motor circuits. SIGNIFICANCE STATEMENT Group Ia proprioceptive sensory neurons form direct synapses with motor neurons, but the molecular mechanisms underlying synapse formation in these monosynaptic sensory–motor connections are unknown. We show that deleting Cdc42 in sensory neurons does not affect proprioceptive sensory axon targeting because axons reach the ventral spinal cord appropriately, but these neurons form significantly fewer presynaptic terminals on motor neurons. Electrophysiological analysis further shows that EPSPs are decreased in these mice. Finally, we demonstrate that Cdc42 is involved in neuroligin-dependent presynaptic differentiation of proprioceptive sensory neurons in vitro

  14. Identification and Characterization of a Chloroplast-Targeted Obg GTPase in Dendrobium officinale.

    Science.gov (United States)

    Chen, Ji; Deng, Feng; Deng, Mengsheng; Han, Jincheng; Chen, Jianbin; Wang, Li; Yan, Shen; Tong, Kai; Liu, Fan; Tian, Mengliang

    2016-12-01

    Bacterial homologous chloroplast-targeted Obg GTPases (ObgCs) belong to the plant-typical Obg group, which is involved in diverse physiological processes during chloroplast development. However, the evolutionarily conserved function of ObgC in plants remains elusive and requires further investigation. In this study, we identified DoObgC from an epiphytic plant Dendrobium officinale and demonstrated the characteristics of DoObgC. Sequence analysis indicated that DoObgC is highly conserved with other plant ObgCs, which contain the chloroplast transit peptide (cTP), Obg fold, G domain, and OCT regions. The C terminus of DoObgC lacking the chloroplast-targeting cTP region, DoObgC Δ1-160 , showed strong similarity to ObgE and other bacterial Obgs. Overexpression of DoObgC Δ1-160 in Escherichia coli caused slow cell growth and an increased number of elongated cells. This phenotype was consistent with the phenotype of cells overexpressing ObgE. Furthermore, the expression of recombinant DoObgC Δ1-160 enhanced the cell persistence of E. coli to streptomycin. Results of transient expression assays revealed that DoObgC was localized to chloroplasts. Moreover, we demonstrated that DoObgC could rescue the embryotic lethal phenotype of the Arabidopsis obgc-t mutant, suggesting that DoObgC is a functional homolog to Arabidopsis AtObgC in D. officinale. Gene expression profiles showed that DoObgC was expressed in leaf-specific and light-dependent patterns and that DoObgC responded to wounding treatments. Our previous and present studies reveal that ObgC has an evolutionarily conserved role in ribosome biogenesis to adapt chloroplast development to the environment.

  15. Concentrações de cera de carnaúba na qualidade de pêssegos cv. Esmeralda armazenados sob refrigeração = Concentrations of carnauba wax on the quality of cv. Esmeralda peaches after storage

    Directory of Open Access Journals (Sweden)

    Marcelo Barbosa Malgarim

    2007-10-01

    wax. ‘Esmeralda’ peaches showed small firmness in the treatments with 75% and 100% of wax. Fruits treated with wax with 50%, 75% and 100% showed small percentage of rottenness. The sensorial evaluation showedthat acidity and astringency in fruits treated with wax decreased. The utilization of 50% wax, associated with cold storage, reduced weight loss and rottenness, maintaining the pulp firmness. Thus, it is possible to store peaches cv. Esmeralda during 30 days at 0ºC and four days at 8ºC.

  16. Peach response to water deficit in a semi-arid region

    Science.gov (United States)

    Paltineanu, C.; Septar, L.; Moale, C.; Nicolae, S.; Nicola, C.

    2013-09-01

    During three years a deficit irrigation experiment was performed on peach response under the semi-arid conditions of south-eastern Romania. Three sprinkler-irrigated treatments were investigated: fully irrigated, deficit irrigation treatment, and non-irrigated control treatment. Soil water content ranged between 60 and 76% of the plant available soil water capacity in fully irrigated, between 40 and 62% in deficit irrigation treatment, and between 30 and 45% in control. There were significant differences in fruit yield between the treatments. Irrigation water use efficiency was maximum in deficit irrigation treatment. Fruit yield correlated significantly with irrigation application. Total dry matter content, total solids content and titrable acidity of fruit were significantly different in the irrigated treatments vs. the control. Significant correlation coefficients were found between some fruit chemical components. For the possible future global warming conditions, when water use becomes increasingly restrictive, deficit irrigation will be a reasonable solution for water conservation in regions with similar soil and climate conditions.

  17. Peach Bottom Atomic Power Station recirc pipe dose rates with zinc injection and condenser replacement

    International Nuclear Information System (INIS)

    DiCello, D.C.; Odell, A.D.; Jackson, T.J.

    1995-01-01

    Peach Bottom Atomic Power Station (PBAPS) is located near the town of Delta, Pennsylvania, on the west bank of the Susquehanna River. It is situated approximately 20 miles south of Lancaster, Pennsylvania. The site contains two boiling water reactors of General Electric design and each rated at 3,293 megawatts thermal. The units are BWR 4s and went commercial in 1977. There is also a decommissioned high temperature gas-cooled reactor on site, Unit 1. PBAPS Unit 2 recirc pipe was replaced in 1985 and Unit 3 recirc pipes replaced in 1988 with 326 NGSS. The Unit 2 replacement pipe was electropolished, and the Unit 3 pipe was electropolished and passivated. The Unit 2 brass condenser was replaced with a Titanium condenser in the first quarter of 1991, and the Unit 3 condenser was replaced in the fourth quarter of 1991. The admiralty brass condensers were the source of natural zinc in both units. Zinc injection was initiated in Unit 2 in May 1991, and in Unit 3 in May 1992. Contact dose rate measurements were made in standard locations on the 28-inch recirc suction and discharge lines to determine the effectiveness of zinc injection and to monitor radiation build-up in the pipe. Additionally, HPGe gamma scans were performed to determine the isotopic composition of the oxide layer inside the pipe. In particular, the specific (μCi/cm 2 ) of Co-60 and Zn-65 were analyzed

  18. Carbohydrate composition of peach palm (Bactris gasipaes Kunth) by-products flours.

    Science.gov (United States)

    Bolanho, Beatriz Cervejeira; Danesi, Eliane Dalva Godoy; Beléia, Adelaide Del Pino

    2015-06-25

    The flours obtained from peach palm by-products are rich in dietary fiber (62-71%) and they can be used as food ingredients. The aim of this work was to investigate the carbohydrate composition of the flours processed from the residual parts (stem and median sheath) of a hearts-of-palm industry. The flours were fractionated, based on their solubility, whose monomeric compounds were determined. The fraction containing mostly cellulose (S5) was the most abundant (26-28%), followed by the sum of fractions (S2, 53, S4) extracted with alkaline solutions (21-22%). The S1 fraction contained the highest percentage of uronic acids, which characterizes the presence of pectin. Xylose and arabinose were found in high proportion in S2 and S3 fractions. The S4 and S5 fractions, rich in glucose, were the main portion of the cell wall material and correspond to the insoluble fraction of the dietary fiber. Copyright © 2015 Elsevier Ltd. All rights reserved.

  19. Identifying yeast isolated from spoiled peach puree and assessment of its batch culture for invertase production

    Directory of Open Access Journals (Sweden)

    Marcela Vega FERREIRA

    Full Text Available Abstract The identification of yeasts isolated from spoiled Jubileu peach puree using the API 20C AUX method and a commercial yeast as witness were studied. Subsequently, the yeast’s growth potential using two batch culture treatments were performed to evaluate number of colonies (N, reducing sugar concentration (RS, free-invertase (FI, and culture-invertase activity (CI. Stock cultures were maintained on potato dextrose agar (PDA slants at 4 °C and pH 5 for later use for batch-culture (150 rpm at 30°C for 24 h, then they were stored at 4 °C for subsequent invertase extraction. The FI extract was obtained using NaHCO3 as autolysis agent, and CI activity was determined on the supernatant after batch-cultured centrifugation. The activity was followed by an increase in absorbance at 490 nm using the acid 3,5-DNS method with glucose standard. Of the four yeasts identified, Saccharomyces cerevisiae was chosen for legal reasons. It showed logarithmic growth up to 18 h of fermentation with positive correlation CI activity and inverse with RS. FI showed greater activity by the end of the log phase and an inverse correlation with CI activity. Finally, it was concluded that treatment “A” is more effective than “B” to produce invertase (EC 3.2.1.26.

  20. Socio-economic impacts of nuclear generating stations: Peach Bottom case study

    International Nuclear Information System (INIS)

    Pijawka, K.D.

    1982-07-01

    This report documents a case study of the socio-economic impacts of the construction and operation of the Peach Bottom nuclear power station. It is part of a major post-licensing study of the socio-economic impacts at twelve nuclear power stations. The case study covers the period beginning with the announcement of plans to construct the reactor and ending in the period 1980 to 1981. The case study deals with changes in the economy, population, settlement patterns and housing, local government and public services, social structure, and public response in the study area during the construction/operation of the reactor. A regional modeling approach is used to trace the impact of the construction/operation on the local economy, labor market, and housing market. Emphasis in the study is on the attribution of socio-economic impacts to the reactor or other causal factors. As part of the study of local public response to the construction/operation of the reactor, the effects of the Three Mile Island accident are examined

  1. Distribution of fission products in Peach Bottom HTGR fuel element E01-01

    International Nuclear Information System (INIS)

    Wichner, R.P.; Dyer, F.F.; Martin, W.J.; Fairchild, L.L.

    1978-10-01

    The fifth in a projected series of six postirradiation examinations of Peach Bottom High-Temperature Gas-Cooled Reactor driver fuel elements is described. The element analyzed received an equivalent of 897 full-power days of irradiation prior to the scheduled termination of Core 2 operation. The examination procedures emphasized the determination of fission product distributions in the graphite portions of the fuel element. Continuous axial scans indicated a 137 Cs inventory of 20.3 Ci in the graphite sleeve and 8.1 Ci in the spine at the time of element withdrawal from the core. In addition, the nuclides 134 Cs, /sup 110 m/Ag, 60 Co, and 154 Eu were found in the graphite portions of the fuel element in significant amounts. Radial distributions of these nuclides plus the beta-emitters 3 H, 14 C, and 90 Sr were obtained at four axial locations of the fueled region of the element sleeve and two axial locations of the element spine. The radial dissection was accomplished by use of a manipulator-operated lathe in a hot cell. In addition to fission product distributions, the appearance of the component parts of the element was recorded photographically, fuel compact and graphite dimensions were recorded at numerous locations, and metallographic examinations of the fuel were performed

  2. Fruit quality in the peach and nectarine with application of hydrogenated cyanamide and mineral oil

    Directory of Open Access Journals (Sweden)

    Sarita Leonel

    Full Text Available This work evaluated the quality of the fruit in peach and nectarine cultivars with and without the application of hydrogenated cyanamide and mineral oil, for two production cycles (2009 and 2010. The experiment was carried out at the School of Agricultural Science of the São Paulo State University (UNESP, at Botucatu in the Brazilian state of São Paulo, located at latitude 22º51'55" S and longitude 48º26'22" E, at an altitude of 810 m. The predominant climate type is warm temperate (mesothermal with rains in the summer and dry in the winter. The following were evaluated: soluble solids, titratable acidity, pH, ratio, firmness, vitamin C and pulp yield. The use of hydrogenated cyanamide and mineral oil had no effect on the quality attributes of the fruit, except for pH, where those fruits under application of the products showed higher values. The cultivars all had a pulp yield greater than 90%, with 'Tourmaline' showing the highest yield (96 %. The levels of vitamin C varied according to the cultivars, where 'Marli' (16.9 mg 100 g-1 and 'Dourado-2' (16.5 mg 100 g-1, stood out for having the highest levels.

  3. Analysis of core damage frequency: Peach Bottom, Unit 2 internal events appendices

    International Nuclear Information System (INIS)

    Kolaczkowski, A.M.; Cramond, W.R.; Sype, T.T.; Maloney, K.J.; Wheeler, T.A.; Daniel, S.L.

    1989-08-01

    This document contains the appendices for the accident sequence analysis of internally initiated events for the Peach Bottom, Unit 2 Nuclear Power Plant. This is one of the five plant analyses conducted as part of the NUREG-1150 effort for the Nuclear Regulatory Commission. The work performed and described here is an extensive reanalysis of that published in October 1986 as NUREG/CR-4550, Volume 4. It addresses comments from numerous reviewers and significant changes to the plant systems and procedures made since the first report. The uncertainty analysis and presentation of results are also much improved, and considerable effort was expended on an improved analysis of loss of offsite power. The content and detail of this report is directed toward PRA practitioners who need to know how the work was done and the details for use in further studies. The mean core damage frequency is 4.5E-6 with 5% and 95% uncertainty bounds of 3.5E-7 and 1.3E-5, respectively. Station blackout type accidents (loss of all ac power) contributed about 46% of the core damage frequency with Anticipated Transient Without Scram (ATWS) accidents contributing another 42%. The numerical results are driven by loss of offsite power, transients with the power conversion system initially available operator errors, and mechanical failure to scram. 13 refs., 345 figs., 171 tabs

  4. Comparative study of the Peach Bottom turbine trip experiment using two different coupled codes approaches

    International Nuclear Information System (INIS)

    Bambara, M.; Bousbia-Salah, A.; D'Auria, F.

    2005-01-01

    Full text of publication follows: In the last years a great concern about the neutron-3D/thermal-hydraulic codes coupling took place. Owing to the improved computational technology, 'best estimate' analyses are today a common tool to assess safety features, and they are necessary if an asymmetric behaviour in the core region exists, or if strong interactions between the core neutronics and reactor thermal-hydraulic occur. In order to validate the coupled codes performances, several international programmes were issued. Among these activities, the OECD/NEA BWR Turbine Trip (TT) was chosen for further sensitivity analyses. It consists of a turbine trip (TT) experiment carried out at the Peach Bottom 2 BWR. In this paper, the results of two different coupled codes systems are summarized and compared. The BWR TT simulations were carried out coupling the thermal-hydraulic system code RELAP5/mode 3.2 to the 3D neutron kinetics code Parcs/2.3, and also the system code ATHLET to the neutronics code QUABOX-CUBBOX. An exhaustive overview of the main features is given, and those aspects, which need further developments and experiences, are pointed out. (authors)

  5. The role of leaves and fruits in determining the specific cultivar characters of peach fruits

    International Nuclear Information System (INIS)

    Manolov, P.; Petrov, A.

    1982-01-01

    At the Institute of Fruit Growing, Plovdiv, triple buds were grafted on the crowns of 6 year-old peach trees in the following cultivar combinations: 1. Springtime (early8 ripening in the second half of June, white fleshed) on Rio oso gem (late, ripening in the first half of September, yellow fleshed); 2. Fillette (early, ripening in the second half of June, yellow fleshed) on Rio oso gem; 3. Rio oso gem on Springtime; 4. Rio oso gem on Fillette. At the begining of the following growing period the development of the grafted generative organs was fully dependent on assimilates produced by the leaves of the other cultivar. The interrelations between the leaves and the fruits in the various combinations were followed by biometrical and radio-isotopic ( 14 C) methods. Results substantiated the conclusion that the genetic information on the development of the specific cultivar characters such as flavour, arome, skin colour, fruit flesh texture and colour, fruit size and date of ripening was borne by the fruits themselves. The synthetic processes of the leaves during photosynthesis are not directly related with the synthetic processes producing the fruits' organic matter. The basic constructing substances were produced in the leaves and were transported to the fruits, where they were subjected to metabolic transformations in accordance with the biological characteristics of the cultivar and the phase of fruit development

  6. Characterization of the vineyard byotyp collection of peach as step in prebreeding process

    Directory of Open Access Journals (Sweden)

    Bakić Ivana

    2016-01-01

    Full Text Available In this research, morphological characterization of vineyard peach collection was conducted. In 78 accessions thirty qualitative traits of tree, flower, leaf, fruit and stone were analysed by using UPOV and ECPGR descriptors. Most of the studied traits showed a high degree of variability. The lowest variability obtained for the leaf traits, and highest for skin and flesh colour. Not only were the accessions sorted into a large number of categories by the most of their properties, but also there was a significant level of variability in the collection, reflecting in the fact that the traits were recombinant in a different way. Principal component analysis (PCA and a dendrogram were performed to determine relationships among accessions and to obtain information on the usefulness of those characters for the discrimination. The PCA revealed that the first 4 principal components were able to represent 43.1% of total variance. Traits with high discriminating values comprised internal and external fruit colour, flower type and colour, flower bud density and stone shape. The cluster analysis showed that the accessions were placed in three main clusters. The greatest impact on the separation in clusters had fruit over colour and extent of fruit over colour. Moreover, the collection can also comprise some accessions with the preferred recombination of properties that might be interesting for further studies in breeding. [Projekat Ministarstva nauke Republike Srbije, br. TR 31063

  7. Effect of Entomopathogenic Nematodes on Mesocriconema xenoplax Populations in Peach and Pecan

    Science.gov (United States)

    Nyczepir, A. P.; Shapiro-Ilan, D. I.; Lewis, E. E.; Handoo, Z. A.

    2004-01-01

    The effect of Steinernema riobrave and Heterorhabditis bacteriophora on population density of Mesocriconema xenoplax in peach was studied in the greenhouse. Twenty-one days after adding 112 M. xenoplax adults and juveniles/1,500 cm³ soil to the soil surface of each pot, 50 infective juveniles/cm² soil surface of either S. riobrave or H. bacteriophora were applied. Another entomopathogenic nematode application of the same density was administered 3 months later. The experiment was repeated once. Mesocriconema xenoplax populations were not suppressed (P ≤ 0.05) in the presence of either S. riobrave or H. bacteriophora 180 days following ring nematode inoculation. On pecan, 200 S. riobrave infective-stage juveniles/cm² were applied to the soil surface of 2-year-old established M. xenoplax populations in field microplots. Additional applications of S. riobrave were administered 2 and 4 months later. This study was terminated 150 days following the initial application of S. riobrave. Populations of M. xenoplax were not suppressed in the presence of S. riobrave. PMID:19262805

  8. Effect of Entomopathogenic Nematodes on Mesocriconema xenoplax Populations in Peach and Pecan.

    Science.gov (United States)

    Nyczepir, A P; Shapiro-Ilan, D I; Lewis, E E; Handoo, Z A

    2004-06-01

    The effect of Steinernema riobrave and Heterorhabditis bacteriophora on population density of Mesocriconema xenoplax in peach was studied in the greenhouse. Twenty-one days after adding 112 M. xenoplax adults and juveniles/1,500 cm(3) soil to the soil surface of each pot, 50 infective juveniles/cm(2) soil surface of either S. riobrave or H. bacteriophora were applied. Another entomopathogenic nematode application of the same density was administered 3 months later. The experiment was repeated once. Mesocriconema xenoplax populations were not suppressed (P

  9. Microsatellite marker analysis of peach-potato aphids (Myzus persicae, Homoptera: Aphididae) from Scottish suction traps.

    Science.gov (United States)

    Malloch, G; Highet, F; Kasprowicz, L; Pickup, J; Neilson, R; Fenton, B

    2006-12-01

    The peach-potato aphid Myzus persicae (Sulzer) is an important vector of plant viruses. A network of suction traps collects aerial samples of this aphid in order to monitor and help predict its spatial distribution and likely impact on virus transmission in crops. A suction trap catch is thought to be a good representation of the total aphid pool. Sensitive molecular markers have been developed that determine the genetic composition of the M. persicae population. In Scotland, UK, these were applied to field collections revealing a limited number of clones. Molecular markers are less successful when applied to specimens that have been preserved in an ethanol-based trap fluid designed to preserve morphology. An assessment of different DNA extraction and PCR techniques is presented and the most efficient are used to analyse M. persicae specimens caught in the Dundee suction trap in 2001, a year when exceptionally high numbers were caught. The results reveal that the majority of the M. persicae caught belonged to two highly insecticide resistant clones. In addition, it was possible to compare the relative frequencies of genotypes caught in the trap with those collected at insecticide treated and untreated field sites in the vicinity. These results indicate that, in addition to suction trap data, the ability to sample field sites provides valuable early warning data which have implications for pest control and virus management strategies.

  10. Enhancing post harvest storage life of peach fruits using calcium chloride

    International Nuclear Information System (INIS)

    Sohail, M.; Afridi, S.R.; Khan, R.U.

    2013-01-01

    Summary: The present study was conducted to enhance the post harvest storage life of peach fruits using calcium chloride treatments. CaCl/sub 2/ solution at 1 % (T1 ), 2 % (T2 ) and 3 % (T3) were prepared and the fruits were dipped for five minutes, while (To) was left with out calcium chloride treatment as control. The fruits were packed in corrugated soft board cartons and stored at ambient temperature (35 degree C + 2) for a total period of 15 days. The physicochemical analysis such as weight loss, fruit firmness, total soluble solids (TSS), decay index, titratable acidity, ascorbic acid content and overall sensory acceptability were determined at an interval of three days up to the successful completion of the study. A significant decrease was observed in fruit firmness (1.8-0.3 kg), % acidity (0.950- 0.538), ascorbic acid content (7.06-4.67 mg/100g) and overall sensory acceptability (8.4-2.3), while a significant increase was observed in TSS (8.2-11.9 degree brix), decay index (0-72.64 %) and % weight loss (0-11.3) during storage. Results showed that one and two percent calcium chloride treated fruits have little improvement while fruits treated with 3 % calcium chloride were found to be most acceptable as per physico-chemical analyses and over all sensory acceptability result. (author)

  11. Postirradiation examination and evaluation of Peach Bottom fuel test element FTE-6

    International Nuclear Information System (INIS)

    Wallroth, C.F.; Holzgraf, J.F.; Jensen, D.D.

    1977-09-01

    Fuel test element FTE-6 was irradiated in the Peach Bottom high-temperature gas-cooled reactor (HTGR) for 645 equivalent full power days. Four fuel varieties, contained in H-327 graphite bodies, were tested. A primary result of this test has been to demonstrate acceptable performance even with calculated high stresses in the graphite bodies. Heterogeneous fuel loadings in the element caused local power peaking and azimuthal power variations, deforming the graphite fuel bodies and thereby causing bowing nearly five times as large as the diametral clearance within the sleeve. The axial stresses resulting from interference between the fuel bodies and sleeve were estimated to have reached 45% of the ultimate material strength at the end of the irradiation. Residual stresses from differential contraction within the fuel body resulted in probable in-plane stress levels of 130% of the material strength at the end-of-life shutdown and of up to 150% of the strength at shutdown during the irradiation cycle. The high in-plane stresses are local peaks at the corners of a sharp notch in the element, which may account for the stresses failing to cause damage. The lack of observable damage, however, indicates that the methods and data used for stress analysis give results that are either fairly accurate or conservative

  12. Monitoring stink bugs (Hemiptera: Pentatomidae) in mid-Atlantic apple and peach orchards.

    Science.gov (United States)

    Leskey, T C; Hogmire, H W

    2005-02-01

    Pyramid traps coated with "industrial safety yellow" exterior latex gloss enamel paint and baited with Euschistus spp. aggregation pheromone, methyl (2E,4Z)-decadienoate captured more stink bugs than all other baited and unbaited trap types in both apple and peach orchards in 2002 and 2003. Commercial sources of dispensers of methyl (2E,4Z)-decadienoate deployed in association with pyramid traps had a significant impact on trap captures. Captures in pyramid traps were four-fold greater when baited with lures from IPM Technologies, Inc. (Portland, OR) than with lures from Suterra (Bend, OR). Variation in yellow pyramid trap color ("industrial safety yellow" and "standard coroplast yellow") and material (plywood, plastic, and masonite) did not affect trap captures. Brown stink bug was the predominant species captured (58%), followed by dusky stink bug, Euschistus tristigmus (Say) (20%); green stink bug, Acrosternum hilare (Say) (14%); and other stink bugs (Brochymena spp. and unidentified nymphs) (8%). Captures in baited pyramid traps were significantly correlated with tree beating samples in both managed and unmanaged apple orchards and with sweep netting samples in the unmanaged apple orchard. However, problems associated with trapping mechanisms of pyramid trap jar tops and jar traps likely resulted in reduced captures in baited traps. Improved trapping mechanisms must be established to develop an effective monitoring tool for stink bugs in mid-Atlantic orchards.

  13. Cytogenetic study on the sterility of peach fruit borer carposina nipponensis (Wals.)

    International Nuclear Information System (INIS)

    Li Yuanying; Kang Wen; Zhang Heqin

    1989-01-01

    The chromosomal aberration, its genetic effect and the structurally abnormal sperms of Peach Fruit Borer (PFB) irradiated with substerilizing dose were described. The numbers of chromosome of PFB were n = 31. The longest chromosome was 4.43 ± 0.49 μ. The shortest one was 1.54 ± 0.15 μ. Total chromosome lengh was 96.37 ± 10.75 μ. The variation rate of the chromosome was 83.6% in F 1 generation including several kinds of aberration. The main chromosomal rearrangement was reciprocal translocation involving many chromosomes. It is the main factor of F 1 generation with higher sterility than P generation that the radiation damage of chromosome can be inherited and strongly expressed in F 1 generation. Because of the dominant lethal mutation of F 1 generation leading to high rate of death, there was less chromosomal aberration in F 2 generation. The variation rate was 12.5%. The genetic abnormalities of ultrastructure of sperm were inherited more intensely in F 1 progeny, produced from the cross of P male adults with the irradiated females

  14. Biochemical Responses of Peach Leaves Infected with Taphrina Deformans Berk/Tul.

    Directory of Open Access Journals (Sweden)

    Lyubka Koleva-Valkova

    2017-01-01

    Full Text Available The phytopathogenic fungus Taphrina deformans causing the so called “leaf curl disease” in peach trees leads to severe yield losses due to the development of leaf hypertrophy and subsequent necrosis and scission. Because of its economic importance, the molecular mechanisms underlying the onset and progression of the disease are of considerable interest to the agricultural science. In this study various biochemical parameters, including the activities of the antioxidant enzymes guaiacol peroxidase, syringaldazine peroxidase and catalase, total polyphenols and anthocyanin content, concentration of free proline, antiradical activity and quantity of plastid pigments, were characterized. All these were measured in both leaves with clear symptoms and distally situated leaves from the same plant that show no signs of the infection. The results demonstrate that the pathogen induces considerable biochemical changes concerning enzymatic and non‑enzymatic elements of the plant defense and antioxidant systems. Moreover, it seems that the fungus provokes a systemic response detectable even in the tissues without observable symptoms.

  15. Peach Bottom Atomic Power Station recirc pipe dose rates with zinc injection and condenser replacement

    Energy Technology Data Exchange (ETDEWEB)

    DiCello, D.C.; Odell, A.D.; Jackson, T.J. [PECO Energy Co., Delta, PA (United States)

    1995-03-01

    Peach Bottom Atomic Power Station (PBAPS) is located near the town of Delta, Pennsylvania, on the west bank of the Susquehanna River. It is situated approximately 20 miles south of Lancaster, Pennsylvania. The site contains two boiling water reactors of General Electric design and each rated at 3,293 megawatts thermal. The units are BWR 4s and went commercial in 1977. There is also a decommissioned high temperature gas-cooled reactor on site, Unit 1. PBAPS Unit 2 recirc pipe was replaced in 1985 and Unit 3 recirc pipes replaced in 1988 with 326 NGSS. The Unit 2 replacement pipe was electropolished, and the Unit 3 pipe was electropolished and passivated. The Unit 2 brass condenser was replaced with a Titanium condenser in the first quarter of 1991, and the Unit 3 condenser was replaced in the fourth quarter of 1991. The admiralty brass condensers were the source of natural zinc in both units. Zinc injection was initiated in Unit 2 in May 1991, and in Unit 3 in May 1992. Contact dose rate measurements were made in standard locations on the 28-inch recirc suction and discharge lines to determine the effectiveness of zinc injection and to monitor radiation build-up in the pipe. Additionally, HPGe gamma scans were performed to determine the isotopic composition of the oxide layer inside the pipe. In particular, the specific ({mu}Ci/cm{sup 2}) of Co-60 and Zn-65 were analyzed.

  16. The evolution of insecticide resistance in the peach potato aphid, Myzus persicae.

    Science.gov (United States)

    Bass, Chris; Puinean, Alin M; Zimmer, Christoph T; Denholm, Ian; Field, Linda M; Foster, Stephen P; Gutbrod, Oliver; Nauen, Ralf; Slater, Russell; Williamson, Martin S

    2014-08-01

    The peach potato aphid, Myzus persicae is a globally distributed crop pest with a host range of over 400 species including many economically important crop plants. The intensive use of insecticides to control this species over many years has led to populations that are now resistant to several classes of insecticide. Work spanning over 40 years has shown that M. persicae has a remarkable ability to evolve mechanisms that avoid or overcome the toxic effect of insecticides with at least seven independent mechanisms of resistance described in this species to date. The array of novel resistance mechanisms, including several 'first examples', that have evolved in this species represents an important case study for the evolution of insecticide resistance and also rapid adaptive change in insects more generally. In this review we summarise the biochemical and molecular mechanisms underlying resistance in M. persicae and the insights study of this topic has provided on how resistance evolves, the selectivity of insecticides, and the link between resistance and host plant adaptation. Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.

  17. An ecological method to understand agricultural standardization in peach orchard ecosystems.

    Science.gov (United States)

    Wan, Nian-Feng; Zhang, Ming-Yi; Jiang, Jie-Xian; Ji, Xiang-Yun; Hao-Zhang

    2016-02-22

    While the worldwide standardization of agricultural production has been advocated and recommended, relatively little research has focused on the ecological significance of such a shift. The ecological concerns stemming from the standardization of agricultural production may require new methodology. In this study, we concentrated on how ecological two-sidedness and ecological processes affect the standardization of agricultural production which was divided into three phrases (pre-, mid- and post-production), considering both the positive and negative effects of agricultural processes. We constructed evaluation indicator systems for the pre-, mid- and post-production phases and here we presented a Standardization of Green Production Index (SGPI) based on the Full Permutation Polygon Synthetic Indicator (FPPSI) method which we used to assess the superiority of three methods of standardized production for peaches. The values of SGPI for pre-, mid- and post-production were 0.121 (Level IV, "Excellent" standard), 0.379 (Level III, "Good" standard), and 0.769 × 10(-2) (Level IV, "Excellent" standard), respectively. Here we aimed to explore the integrated application of ecological two-sidedness and ecological process in agricultural production. Our results are of use to decision-makers and ecologists focusing on eco-agriculture and those farmers who hope to implement standardized agricultural production practices.

  18. Yersinia outer protein YopE affects the actin cytoskeleton in Dictyostelium discoideum through targeting of multiple Rho family GTPases

    LENUS (Irish Health Repository)

    Vlahou, Georgia

    2009-07-14

    Abstract Background All human pathogenic Yersinia species share a virulence-associated type III secretion system that translocates Yersinia effector proteins into host cells to counteract infection-induced signaling responses and prevent phagocytosis. Dictyostelium discoideum has been recently used to study the effects of bacterial virulence factors produced by internalized pathogens. In this study we explored the potential of Dictyostelium as model organism for analyzing the effects of ectopically expressed Yersinia outer proteins (Yops). Results The Yersinia pseudotuberculosis virulence factors YopE, YopH, YopM and YopJ were expressed de novo within Dictyostelium and their effects on growth in axenic medium and on bacterial lawns were analyzed. No severe effect was observed for YopH, YopJ and YopM, but expression of YopE, which is a GTPase activating protein for Rho GTPases, was found to be highly detrimental. GFP-tagged YopE expressing cells had less conspicuous cortical actin accumulation and decreased amounts of F-actin. The actin polymerization response upon cAMP stimulation was impaired, although chemotaxis was unaffected. YopE also caused reduced uptake of yeast particles. These alterations are probably due to impaired Rac1 activation. We also found that YopE predominantly associates with intracellular membranes including the Golgi apparatus and inhibits the function of moderately overexpressed RacH. Conclusion The phenotype elicited by YopE in Dictyostelium can be explained, at least in part, by inactivation of one or more Rho family GTPases. It further demonstrates that the social amoeba Dictyostelium discoideum can be used as an efficient and easy-to-handle model organism in order to analyze the function of a translocated GAP protein of a human pathogen.

  19. Expression of GIMAP1, a GTPase of the immunity-associated protein family, is not up-regulated in malaria

    Directory of Open Access Journals (Sweden)

    Carter Christine

    2009-04-01

    Full Text Available Abstract Background GIMAP (GTPase of the immunity-associated protein family proteins are a family of putative GTPases believed to be regulators of cell death in lymphomyeloid cells. GIMAP1 was the first reported member of this gene family, identified as a gene up-regulated at the RNA level in the spleens of mice infected with the malarial parasite, Plasmodium chabaudi. Methods A monoclonal antibody against mouse GIMAP1 was developed and was used to analyse the expression of the endogenous protein in tissues of normal mice and in defined sub-populations of cells prepared from lymphoid tissues using flow cytometry. It was also used to assess the expression of GIMAP1 protein after infection and/or immunization of mice with P. chabaudi. Real-time PCR analysis was employed to measure the expression of GIMAP1 for comparison with the protein level analysis. Results GIMAP1 protein expression was detected in all lineages of lymphocytes (T, B, NK, in F4/80+ splenic macrophages and in some lymphoid cell lines. Additional evidence is presented suggesting that the strong expression by mature B cells of GIMAP1 and other GIMAP genes and proteins seen in mice may be a species-dependent characteristic. Unexpectedly, no increase was found in the expression of GIMAP1 in P. chabaudi infected mice at either the mRNA or protein level, and this remained so despite applying a number of variations to the protocol. Conclusion The model of up-regulation of GIMAP1 in response to infection/immunization with P. chabaudi is not a robustly reproducible experimental system. The GIMAP1 protein is widely expressed in lymphoid cells, with an interesting increase in expression in the later stages of B cell development. Alternative approaches will be required to define the functional role of this GTPase in immune cells.

  20. Increased diacylglycerol kinase ζ expression in human metastatic colon cancer cells augments Rho GTPase activity and contributes to enhanced invasion

    International Nuclear Information System (INIS)

    Cai, Kun; Mulatz, Kirk; Ard, Ryan; Nguyen, Thanh; Gee, Stephen H

    2014-01-01

    Unraveling the signaling pathways responsible for the establishment of a metastatic phenotype in carcinoma cells is critically important for understanding the pathology of cancer. The acquisition of cell motility is a key property of metastatic tumor cells and is a prerequisite for invasion. Rho GTPases regulate actin cytoskeleton reorganization and the cellular responses required for cell motility and invasion. Diacylglycerol kinase ζ (DGKζ), an enzyme that phosphorylates diacylglycerol to yield phosphatidic acid, regulates the activity of the Rho GTPases Rac1 and RhoA. DGKζ mRNA is highly expressed in several different colon cancer cell lines, as well as in colon cancer tissue relative to normal colonic epithelium, and thus may contribute to the metastatic process. To investigate potential roles of DGKζ in cancer metastasis, a cellular, isogenic model of human colorectal cancer metastatic transition was used. DGKζ protein levels, Rac1 and RhoA activity, and PAK phosphorylation were measured in the non-metastatic SW480 adenocarcinoma cell line and its highly metastatic variant, the SW620 line. The effect of DGKζ silencing on Rho GTPase activity and invasion through Matrigel-coated Transwell inserts was studied in SW620 cells. Invasiveness was also measured in PC-3 prostate cancer and MDA-MB-231 breast cancer cells depleted of DGKζ. DGKζ protein levels were elevated approximately 3-fold in SW620 cells compared to SW480 cells. There was a concomitant increase in active Rac1 in SW620 cells, as well as substantial increases in the expression and phosphorylation of the Rac1 effector PAK1. Similarly, RhoA activity and expression were increased in SW620 cells. Knockdown of DGKζ expression in SW620 cells by shRNA-mediated silencing significantly reduced Rac1 and RhoA activity and attenuated the invasiveness of SW620 cells in vitro. DGKζ silencing in highly metastatic MDA-MB-231 breast cancer cells and PC-3 prostate cancer cells also significantly attenuated

  1. The Rho GTPase Effector ROCK Regulates Cyclin A, Cyclin D1, and p27Kip1 Levels by Distinct Mechanisms

    OpenAIRE

    Croft, Daniel R.; Olson, Michael F.

    2006-01-01

    The members of the Rho GTPase family are well known for their regulation of actin cytoskeletal structures. In addition, they influence progression through the cell cycle. The RhoA and RhoC proteins regulate numerous effector proteins, with a central and vital signaling role mediated by the ROCK I and ROCK II serine/threonine kinases. The requirement for ROCK function in the proliferation of numerous cell types has been revealed by studies utilizing ROCK-selective inhibitors such as Y-27632. H...

  2. Development of a Novel NMR-based Rheb GTPase Assay and Molecular Characterization of TSC2 GAP Activity

    Science.gov (United States)

    2010-05-01

    GTPase) that belongs to the Ras superfamily and has homologs in yeast, fungi , slime mold, fruit fly, zebra fish, and mammals (1–3). Ge- netic and...characterization of TSC2 disease mutations affecting its GAP activity (months 9-12) While the final aspects of this task are yet to be completed, we have...domain mutants of TSC2 that we examined affected its enzymatic activ- ity. This method can now be applied to study the function and regulation of other

  3. Hypersensitivity to Tomato (Lycopersicon esculentum) in Peach-Allergic Patients: rPrup 3 and rPrup 1 Are Predictive of Symptom Severity.

    Science.gov (United States)

    Mascheri, A; Farioli, L; Pravettoni, V; Piantanida, M; Stafylaraki, C; Scibilia, J; Mirone, C; Preziosi, D; Nichelatti, M; Pastorello, E A

    2015-01-01

    Background: The role of allergens in the severity of tomato allergy symptoms has not yet been studied. To evaluate the relationship between severe allergic reactions to peach and tomato and between tomato allergy symptoms and the pattern of IgE positivity for rPru p 1, rPru p 3, rPru p 4, rBetv 1, rBetv 2, rBetv4, rPhl p 1, and rPhl p 12 in order to identify the role of recombinant allergens in the severity of reactions to tomato. We studied peach-allergic patients with clinical reactions to tomato by performing an open food challenge, skin prick test, and determination of serum specific IgE to tomato and to recombinant peach, birch, and grass allergens. Statistical analysis was carried out to evaluate the relationship between the severity of tomato symptoms and IgE positivity to the different allergens and to peach-induced symptoms. We found a significant association between severe reactions to tomato and severe reactions to peach (P = .01 7) and levels of IgE to rPru p3 (P = .029) and between mild tomato allergy symptoms and levels of IgE to rPru p1 (P = .047), anti-rBetv 1 (P = .0414), anti-rBetv 2 (P = .0457), and Phleum pratense (P = .0022). We observed a significant relationship between peach and symptoms of tomato allergy. IgE positivity for rPru p3 seems to be a surrogate biochemical marker for severe tomato allergy, whereas the presence of anti-rPru p 1 IgE may be an indicator of mild tomato allergy.

  4. Phytohormone Signaling of the Resistance to Plum pox virus (PPV, Sharka Disease) Induced by Almond (Prunus dulcis (Miller) Webb) Grafting to Peach (P. persica L. Batsch)

    OpenAIRE

    Azam Nikbakht Dehkordi; Manuel Rubio; Nadali Babaeian; Alfonso Albacete; Pedro Martínez-Gómez

    2018-01-01

    Plum pox virus (PPV, sharka) is a limiting factor for peach production, and no natural sources of resistance have been described. Recent studies, however, have demonstrated that grafting the almond cultivar “Garrigues” onto the “GF305” peach infected with Dideron-type (PPV-D) isolates progressively reduces disease symptoms and virus accumulation. Furthermore, grafting “Garrigues” onto “GF305” prior to PPV-D inoculation has been found to completely prevent virus infection, showing that resista...

  5. Molecular cloning of plane pollen allergen Pla a 3 and its utility as diagnostic marker for peach associated plane pollen allergy.

    Science.gov (United States)

    Wangorsch, A; Larsson, H; Messmer, M; García-Moral, A; Lauer, I; Wolfheimer, S; Schülke, S; Bartra, J; Vieths, S; Lidholm, J; Scheurer, S

    2016-05-01

    Non-specific lipid transfer proteins (nsLTP) are considered to provoke allergic symptoms to plane tree pollen, which are frequently associated with peach allergy. The objective was to clone the cDNA of plane pollen nsLTP Pla a 3, to characterize IgE-binding and allergenic potency of recombinant Pla a 3 in comparison to its natural counterpart and peach nsLTP Pru p 3. Natural Pla a 3 was purified from plane pollen and analysed by mass spectrometry (MS). Recombinant Pla a 3 was characterized by SDS-PAGE and CD spectroscopy. Specific IgE to extract, components of plane pollen and Pru p 3 was measured by ImmunoCAP in sera of patients allergic to either plane pollen (n = 10), peach (n = 15) or both (n = 15). Biological potency of the proteins was investigated by in vitro mediator release assays and IgE cross-reactivity by competitive ELISA. Two Pla a 3 isoforms were identified. Recombinant Pla a 3 showed high purity, structural integrity, IgE-binding capacity comparable to nPla a 3 and biological potency. Sensitization to plane pollen extract was confirmed in 24/25 plane pollen allergics. The frequency of sensitization to Pla a 3 was 53% among patients allergic to both plane pollen and peach and 10% among plane pollen allergics tolerating peach where most patients were sensitized to Pla a 1. Pla a 3 and Pru p 3 showed strong bi-directional IgE cross-reactivity in patients allergic to peach and plane pollen, but not in peach allergics tolerating plane pollen. Levels of IgE-binding were generally higher to Pru p 3 than to Pla a 3. Sensitization to Pla a 3 is relevant in a subgroup of plane pollen allergics with concomitant peach allergy. IgE testing with Pla a 3 may serve as a marker to identify plane pollen allergic patients at risk of LTP-mediated food reactions and thereby improve in vitro diagnostic procedures. © 2016 John Wiley & Sons Ltd.

  6. Multivariate analysis applied to the study of the relationship between soil and plant properties in a peach orchard

    Directory of Open Access Journals (Sweden)

    Viviane Santos Silva Terra

    2014-06-01

    Full Text Available In the State of Rio Grande do Sul, the municipality of Pelotas is responsible for 90 % of peach production due to its suitable climate and soil conditions. However, there is the need for new studies that aim at improved fruit quality and increased yield. The aim of this study was to evaluate the relationship that exists between soil physical properties and properties in the peach plant in the years 2010 and 2011 by the technique of multivariate canonical correlation. The experiment was conducted in a peach orchard located in the municipality of Morro Redondo, RS, Brazil, where an experimental grid of 101 plants was established. In a trench dug beside each one of the 101 plants, soil samples were collected to determine silt, clay, and sand contents, soil density, total porosity, macroporosity, microporosity, and volumetric water content in the 0.00-0.10 and 0.10-0.20 m layers, as well as the depth of the A horizon. In each plant and in each year, the following properties were assessed: trunk diameter, fruit size and number of fruits per plant, average weight of the fruit per plant, fruit pulp firmness, Brix content, and yield from the orchard. Exploratory analysis of the data was undertaken by descriptive statistics, and the relationships between the physical properties of the soil and of the plant were assessed by canonical correlation analysis. The results showed that the clay and microporosity variables were those that exhibited the highest coefficients of canonical cross-loading with the plant properties in the soil layers assessed, and that the variable of mean weight of the fruit per plant was that which had the highest coefficients of canonical loading within the plant group for the two years assessed.

  7. Disruption of Ethylene Responses by Turnip mosaic virus Mediates Suppression of Plant Defense against the Green Peach Aphid Vector.

    Science.gov (United States)

    Casteel, Clare L; De Alwis, Manori; Bak, Aurélie; Dong, Haili; Whitham, Steven A; Jander, Georg

    2015-09-01

    Plants employ diverse responses mediated by phytohormones to defend themselves against pathogens and herbivores. Adapted pathogens and herbivores often manipulate these responses to their benefit. Previously, we demonstrated that Turnip mosaic virus (TuMV) infection suppresses callose deposition, an important plant defense induced in response to feeding by its aphid vector, the green peach aphid (Myzus persicae), and increases aphid fecundity compared with uninfected control plants. Further, we determined that production of a single TuMV protein, Nuclear Inclusion a-Protease (NIa-Pro) domain, was responsible for changes in host plant physiology and increased green peach aphid reproduction. To characterize the underlying molecular mechanisms of this phenomenon, we examined the role of three phytohormone signaling pathways, jasmonic acid, salicylic acid, and ethylene (ET), in TuMV-infected Arabidopsis (Arabidopsis thaliana), with or without aphid herbivory. Experiments with Arabidopsis mutants ethylene insensitive2 and ethylene response1, and chemical inhibitors of ET synthesis and perception (aminoethoxyvinyl-glycine and 1-methylcyclopropene, respectively), show that the ET signaling pathway is required for TuMV-mediated suppression of Arabidopsis resistance to the green peach aphid. Additionally, transgenic expression of NIa-Pro in Arabidopsis alters ET responses and suppresses aphid-induced callose formation in an ET-dependent manner. Thus, disruption of ET responses in plants is an additional function of NIa-Pro, a highly conserved potyvirus protein. Virus-induced changes in ET responses may mediate vector-plant interactions more broadly and thus represent a conserved mechanism for increasing transmission by insect vectors across generations. © 2015 American Society of Plant Biologists. All Rights Reserved.

  8. Evaluation of different doses of gamma radiation on physicochemical characteristics of peach Prunus persica (cv. Chimarrita) minimally processed

    International Nuclear Information System (INIS)

    Oliveira, Ana Claudia S.; Silva, Lucia C.A.S.; Perecin, Thalita Neme; Arthur, Valter; Harder, Marcia N.C.; Mansi, Debora N.; Canniatti-Brazaca, Solange G.

    2009-01-01

    The objective was to evaluate the effect of different doses of gamma radiation on the physico-chemical characteristics of peach Prunus persica (cv. Chimarrita) minimally processed, to increase the useful life of the fruit. The peaches were purchased at Ceasa of Campinas/SP and taken to the Laboratory of Radiobiology and Environment of CENA/USP (Piracicaba/SP), which were washed in tap water, peeled and cut into four pieces. The pieces of peach were dipped in sodium hypochlorite solution of 15 mL/L for 4 minutes and dry in a plastic support. Then it were placed in plastic containers (polypropylene). Subsequently, they were irradiated in a Cobalt-60 source, type Gammacell-220 (dose rate of 0,543 kGy/hour) with doses of: 0 (control), 1.0 and 2.0 kGy and stored at a temperature of 8 deg C. The experimental was developed entirely at random with 3 replicates for each treatment. For the statistic analysis was using the Tuckey test at 5% level of probability. Subsequently, analysis was carried out: color factors (l, a, b), pH, soluble solids (deg Brix), acidity and vitamin C. The tests were performed at 1, 3 and 6 days after irradiation. According to the results concluded that the analysis of color and acidity there was no significant difference between treatments, however, for the soluble solids (deg Brix), vitamin C and texture significant difference showing a decrease proportional to increasing doses of radiation and storage time. But the pH increased in relation to dose and during the analysis. (author)

  9. Comparative RNA-seq analysis of early-infected peach leaves by the invasive phytopathogen Xanthomonas arboricola pv. pruni.

    Directory of Open Access Journals (Sweden)

    Didier Socquet-Juglard

    Full Text Available Xanthomonas arboricola pv. pruni is a quarantine bacterial pathogen that threatens peach production by causing necrotic spots on leaves and fruits, thus with the potential of severely reducing yields. The current understanding of the host plant defense responses to the pathogen is very limited. Using whole transcriptome sequencing, differential gene expression was analyzed at two time points, 2 h and 12 h post inoculation (hpi, by comparing the inoculated samples to their respective controls. On the total of 19,781 known peach genes that were expressed in all time points and conditions, 34 and 263 were differentially expressed at 2 and 12 hpi, respectively. Of those, 82% and 40% were up-regulated, respectively; and 18% and 60% were down-regulated, respectively. The functional annotation based on gene ontology (GO analysis highlighted that genes involved in metabolic process and response to stress were particularly represented at 2 hpi whereas at 12 hpi cellular and metabolic processes were the categories with the highest number of genes differentially expressed. Of particular interest among the differentially expressed genes identified were several pathogen-associated molecular pattern (PAMP receptors, disease resistance genes including several RPM1-like and pathogenesis related thaumatin encoding genes. Other genes involved in photosynthesis, in cell wall reorganization, in hormone signaling pathways or encoding cytochrome were also differentially expressed. In addition, novel transcripts were identified, providing another basis for further characterization of plant defense-related genes. Overall, this study gives a first insight of the peach defense mechanisms during the very early stages of infection with a bacterial disease in the case of a compatible interaction.

  10. Radioactivity distribution of the fruit trees ascribable to radioactive fall our (4). Caesium content and its distribution in peach trees

    International Nuclear Information System (INIS)

    Takata, Daisuke; Yasunaga, Eriko; Sasaki, Haruo; Tanoi, Keitaro; Nakanishi, Tomoko M.; Oshita, Seiichi

    2012-01-01

    Movement and distribution of radiocaesium in peach tree was studied. The radiocaesium distributed on branches most, which branches were 1, 2 or 3 years old. With the observation of tissues of trunk, the highest radiocaesium concentration was measured at the bark, which was higher than that of soil surface. The radiocaesium concentration was drastically low in the wood part. However, the total amount of radiocaesium of wood was as the same level as that of the bark. About 20% of radiocaesium in the tree was estimated to be removed as fruits and leaves. (author)

  11. Activation of Rho GTPases by Cytotoxic Necrotizing Factor 1 Induces Macropinocytosis and Scavenging Activity in Epithelial Cells

    Science.gov (United States)

    Fiorentini, Carla; Falzano, Loredana; Fabbri, Alessia; Stringaro, Annarita; Logozzi, Mariaantonia; Travaglione, Sara; Contamin, Stéphanette; Arancia, Giuseppe; Malorni, Walter; Fais, Stefano

    2001-01-01

    Macropinocytosis, a ruffling-driven process that allows the capture of large material, is an essential aspect of normal cell function. It can be either constitutive, as in professional phagocytes where it ends with the digestion of captured material, or induced, as in epithelial cells stimulated by growth factors. In this case, the internalized material recycles back to the cell surface. We herein show that activation of Rho GTPases by a bacterial protein toxin, the Escherichia coli cytotoxic necrotizing factor 1 (CNF1), allowed epithelial cells to engulf and digest apoptotic cells in a manner similar to that of professional phagocytes. In particular, we have demonstrated that 1) the activation of all Rho, Rac, and Cdc42 by CNF1 was essential for the capture and internalization of apoptotic cells; and 2) such activation allowed the discharge of macropinosomal content into Rab7 and lysosomal associated membrane protein-1 acidic lysosomal vesicles where the ingested particles underwent degradation. Taken together, these findings indicate that CNF1-induced “switching on” of Rho GTPases may induce in epithelial cells a scavenging activity, comparable to that exerted by professional phagocytes. The activation of such activity in epithelial cells may be relevant, in mucosal tissues, in supporting or integrating the scavenging activity of resident macrophages. PMID:11452003

  12. The interaction properties of the human Rab GTPase family--comparative analysis reveals determinants of molecular binding selectivity.

    Directory of Open Access Journals (Sweden)

    Matthias Stein

    Full Text Available Rab GTPases constitute the largest subfamily of the Ras protein superfamily. Rab proteins regulate organelle biogenesis and transport, and display distinct binding preferences for effector and activator proteins, many of which have not been elucidated yet. The underlying molecular recognition motifs, binding partner preferences and selectivities are not well understood.Comparative analysis of the amino acid sequences and the three-dimensional electrostatic and hydrophobic molecular interaction fields of 62 human Rab proteins revealed a wide range of binding properties with large differences between some Rab proteins. This analysis assists the functional annotation of Rab proteins 12, 14, 26, 37 and 41 and provided an explanation for the shared function of Rab3 and 27. Rab7a and 7b have very different electrostatic potentials, indicating that they may bind to different effector proteins and thus, exert different functions. The subfamily V Rab GTPases which are associated with endosome differ subtly in the interaction properties of their switch regions, and this may explain exchange factor specificity and exchange kinetics.We have analysed conservation of sequence and of molecular interaction fields to cluster and annotate the human Rab proteins. The analysis of three dimensional molecular interaction fields provides detailed insight that is not available from a sequence-based approach alone. Based on our results, we predict novel functions for some Rab proteins and provide insights into their divergent functions and the determinants of their binding partner selectivity.

  13. Potential involvement of drought-induced Ran GTPase CLRan1 in root growth enhancement in a xerophyte wild watermelon.

    Science.gov (United States)

    Akashi, Kinya; Yoshimura, Kazuya; Kajikawa, Masataka; Hanada, Kouhei; Kosaka, Rina; Kato, Atsushi; Katoh, Akira; Nanasato, Yoshihiko; Tsujimoto, Hisashi; Yokota, Akiho

    2016-10-01

    Enhanced root growth is known as the survival strategy of plants under drought. Previous proteome analysis in drought-resistant wild watermelon has shown that Ran GTPase, an essential regulator of cell division and proliferation, was induced in the roots under drought. In this study, two cDNAs were isolated from wild watermelon, CLRan1 and CLRan2, which showed a high degree of structural similarity with those of other plant Ran GTPases. Quantitative RT-PCR and promoter-GUS assays suggested that CLRan1 was expressed mainly in the root apex and lateral root primordia, whereas CLRan2 was more broadly expressed in other part of the roots. Immunoblotting analysis confirmed that the abundance of CLRan proteins was elevated in the root apex region under drought stress. Transgenic Arabidopsis overexpressing CLRan1 showed enhanced primary root growth, and the growth was maintained under osmotic stress, indicating that CLRan1 functions as a positive factor for maintaining root growth under stress conditions.

  14. Fission product behaviour during operation of the second Peach Bottom core

    International Nuclear Information System (INIS)

    Malinauskas, A.P.; Nordwall, H.J. de; Dyer, F.F.; Wichner, R.P.; Martin, W.J.; Kolb, J.O.

    1976-01-01

    The Peach Bottom high-temperature, gas-cooled reactor began operation on 1 June 1967 and continued power production until 9 October 1969, accumulating 452 equivalent full power days (EFPD) operation. After reload, power production with Core 2 began 14 July 1970 and terminated 31 October 1974 after 897 EFPD operation. Surveillance of fission product release and behaviour was intensified during Core 2 operation to permit a wider range of measurements to be made. In addition to monitoring the noble gas content of the fuel element purge system and the coolant circuit, the programme was extended to include measurements of radioactive and other condensible species (including dust) entering or exiting the core and steam generator, and of surface concentrations of gamma-emitting nuclides deposited on the primary coolant surfaces. These data, which were obtained over the operating period April 1971 - October 1974, are summarized and discussed. The data demonstrate that caesium behaviour in the coolant circuit during the first two-thirds of Core 2 life was primarily governed by caesium released during Core 1 operation. The data also indicate that whereas the steam generator surfaces attenuate molecular caesium concentrations in the coolant, the dust-borne component is remarkably persistent. Driver fuel elements were removed from the reactor after 385 EFPD, 701 EFPD, and at end-of-life. These fuel elements are at various stages of an intensive post-irradiation examination. Some of the axial and radial concentration profiles of fission products which have been obtained are likewise presented. Although these profiles indicate varied fission product behaviour, the observations can in general be qualitatively described on the basis of the operational histories of the fuel elements. (author)

  15. The complete nucleotide sequence of RNA 3 of a peach isolate of Prunus necrotic ringspot virus.

    Science.gov (United States)

    Hammond, R W; Crosslin, J M

    1995-04-01

    The complete nucleotide sequence of RNA 3 of the PE-5 peach isolate of Prunus necrotic ringspot ilarvirus (PNRSV) was obtained from cloned cDNA. The RNA sequence is 1941 nucleotides and contains two open reading frames (ORFs). ORF 1 consisted of 284 amino acids with a calculated molecular weight of 31,729 Da and ORF 2 contained 224 amino acids with a calculated molecular weight of 25,018 Da. ORF 2 corresponds to the coat protein gene. Expression of ORF 2 engineered into a pTrcHis vector in Escherichia coli results in a fusion polypeptide of approximately 28 kDa which cross-reacts with PNRSV polyclonal antiserum. Analysis of the coat protein amino acid sequence reveals a putative "zinc-finger" domain at the amino-terminal portion of the protein. Two tetranucleotide AUGC motifs occur in the 3'-UTR of the RNA and may function in coat protein binding and genome activation. ORF 1 homologies to other ilarviruses and alfalfa mosaic virus are confined to limited regions of conserved amino acids. The translated amino acid sequence of the coat protein gene shows 92% similarity to one isolate of apple mosaic virus, a closely related member of the ilarvirus group of plant viruses, but only 66% similarity to the amino acid sequence of the coat protein gene of a second isolate. These relationships are also reflected at the nucleotide sequence level. These results in one instance confirm the close similarities observed at the biophysical and serological levels between these two viruses, but on the other hand call into question the nomenclature used to describe these viruses.

  16. Ascorbic Acid Alleviates Water Stress in Young Peach Trees and Improves Their Performance after Rewatering

    Directory of Open Access Journals (Sweden)

    Consuelo Penella

    2017-09-01

    Full Text Available Exogenous application of biochemicals has been found to improve water stress tolerance in herbaceous crops but there are limited studies on deciduous fruit trees. The goal of this research was to study if ascorbic acid applications could improve physiological mechanisms associated with water stress tolerance in young fruit trees. Ascorbic acid was foliarly applied at a concentration of 250 ppm to water-stressed and well-watered peach trees (control of two cultivars (‘Scarletprince’ and ‘CaroTiger’. Trees received either one or two applications, and 1 week after the second application all trees were rewatered to field capacity. Upon rewatering, CO2 assimilation and stomatal conductance of water-stressed ‘Scarletprince’ trees sprayed with ascorbic acid (one or two applications were similar to those of well-irrigated trees, but water-stressed trees that had not received ascorbic acid did not recover photosynthetical functions. Also, water status in sprayed water-stressed ‘Scarletprince’ trees was improved to values similar to control trees. On the other hand, water-stressed ‘CaroTiger’ trees needed two applications of ascorbic acid to reach values of CO2 assimilation similar to control trees but these applications did not improve their water status. In general terms, different response mechanisms to cope with water stress in presence of ascorbic acid were found in each cultivar, with ‘Scarletprince’ trees preferentially using proline as compatible solute and ‘CaroTiger’ trees relying on stomatal regulation. The application of ascorbic acid reduced cell membrane damage and increased catalase activity in water-stressed trees of both cultivars. These results suggest that foliar applications of ascorbic acid could be used as a management practice for improving water stress tolerance of young trees under suboptimal water regimes.

  17. Ascorbic Acid Alleviates Water Stress in Young Peach Trees and Improves Their Performance after Rewatering.

    Science.gov (United States)

    Penella, Consuelo; Calatayud, Ángeles; Melgar, Juan C

    2017-01-01

    Exogenous application of biochemicals has been found to improve water stress tolerance in herbaceous crops but there are limited studies on deciduous fruit trees. The goal of this research was to study if ascorbic acid applications could improve physiological mechanisms associated with water stress tolerance in young fruit trees. Ascorbic acid was foliarly applied at a concentration of 250 ppm to water-stressed and well-watered peach trees (control) of two cultivars ('Scarletprince' and 'CaroTiger'). Trees received either one or two applications, and 1 week after the second application all trees were rewatered to field capacity. Upon rewatering, CO 2 assimilation and stomatal conductance of water-stressed 'Scarletprince' trees sprayed with ascorbic acid (one or two applications) were similar to those of well-irrigated trees, but water-stressed trees that had not received ascorbic acid did not recover photosynthetical functions. Also, water status in sprayed water-stressed 'Scarletprince' trees was improved to values similar to control trees. On the other hand, water-stressed 'CaroTiger' trees needed two applications of ascorbic acid to reach values of CO 2 assimilation similar to control trees but these applications did not improve their water status. In general terms, different response mechanisms to cope with water stress in presence of ascorbic acid were found in each cultivar, with 'Scarletprince' trees preferentially using proline as compatible solute and 'CaroTiger' trees relying on stomatal regulation. The application of ascorbic acid reduced cell membrane damage and increased catalase activity in water-stressed trees of both cultivars. These results suggest that foliar applications of ascorbic acid could be used as a management practice for improving water stress tolerance of young trees under suboptimal water regimes.

  18. Transcriptome reprogramming of resistant and susceptible peach genotypes during Xanthomonas arboricola pv. pruni early leaf infection.

    Directory of Open Access Journals (Sweden)

    Fabio Gervasi

    Full Text Available Bacterial spot caused by Xanthomonas arboricola pv. pruni (Xap is a major threat to Prunus species worldwide. The molecular mechanisms of peach resistance to Xap during early leaf infection were investigated by RNA-Seq analysis of two Prunus persica cultivars, 'Redkist' (resistant, and 'JH Hale' (susceptible at 30 minutes, 1 and 3 hours-post-infection (hpi. Both cultivars exhibited extensive modulation of gene expression at 30 mpi, which reduced significantly at 1 hpi, increasing again at 3 hpi. Overall, 714 differentially expressed genes (DEGs were detected in 'Redkist' (12% at 30 mpi and 1 hpi and 88% at 3 hpi. In 'JH Hale', 821 DEGs were identified (47% at 30 mpi and 1 hpi and 53% at 3 hpi. Highly up-regulated genes (fold change > 100 at 3 hpi exhibited higher fold change values in 'Redkist' than in 'JH Hale'. RNA-Seq bioinformatics analyses were validated by RT-qPCR. In both cultivars, DEGs included genes with putative roles in perception, signal transduction, secondary metabolism, and transcription regulation, and there were defense responses in both cultivars, with enrichment for the gene ontology terms, 'immune system process', 'defense response', and 'cell death'. There were particular differences between the cultivars in the intensity and kinetics of modulation of expression of genes with putative roles in transcriptional activity, secondary metabolism, photosynthesis, and receptor and signaling processes. Analysis of differential exon usage (DEU revealed that both cultivars initiated remodeling their transcriptomes at 30 mpi; however, 'Redkist' exhibited alternative exon usage for a greater number of genes at every time point compared with 'JH Hale'. Candidate resistance genes (WRKY-like, CRK-like, Copper amine oxidase-like, and TIR-NBS-LRR-like are of interest for further functional characterization with the aim of elucidating their role in Prunus spp. resistance to Xap.

  19. Transgenerational shifts in reproduction hormesis in green peach aphid exposed to low concentrations of imidacloprid.

    Directory of Open Access Journals (Sweden)

    Murali-Mohan Ayyanath

    Full Text Available Hormesis is a biphasic phenomenon that in toxicology is characterized by low-dose stimulation and high-dose inhibition. It has been observed in a wide range of organisms in response to many chemical stressors, including insects exposed to pesticides, with potential repercussions for agriculture and pest management. To address questions related to the nature of the dose-response and potential consequences on biological fitness, we examined transgenerational hormesis in the green peach aphid, Myzus persicae, when exposed to sublethal concentrations of the insecticide imidacloprid. A hormetic response in the form of increased reproduction was consistently observed and a model previously developed to test for hormesis adequately fit some of our data. However, the nature of the dose-response differed within and across generations depending upon the duration and mode of exposure. Decreased reproduction in intermediate generations confirmed that fitness tradeoffs were a consequence of the hormetic response. However, recovery to levels of reproduction equal to that of controls in subsequent generations and significantly greater total reproduction after four generations suggested that biological fitness was increased by exposure to low concentrations of the insecticide, even when insects were continuously exposed to the stressor. This was especially evident in a greenhouse experiment where the instantaneous rate of population increase almost doubled and total aphid production more than quadrupled when aphids were exposed to potato plants systemically treated with low amounts of imidacloprid. Our results show that although fitness tradeoffs do occur with hormetic responses, this does not necessarily compromise overall biological fitness.

  20. Canopy structure and physiology related to rootstock vigour in early-ripening peach cultivar Flordastar

    International Nuclear Information System (INIS)

    Motisi, A.; Grutta, I.; Pernice, F.; Caruso, T.

    2005-01-01

    Canopy architectural and eco-physiological traits were measured on five-year-old early-ripening peach cv Flordastar trees grafted on GF 677 and MrS 2/5 rootstocks. Data are reported both on measurements performed directly on the trees, for branches and twigs characters, and on the fractal dimension (D), estimated by the 'box counting' method taken from digital images of Winter-dormant trees, adopted as an indicator of canopy complexity. Results are discussed in relation to the modification of the canopy microclimate as a consequence of the effects of rootstock on tree architecture and water consumption, the latter measured by using sap flow (HPV) probes. A lower degree of canopy complexity was observed in trees grafted onto MrS 2/5 and this, in turn, was related to a higher degree of aerodynamic contact of the tree with the atmosphere (expressed in terms of leaf boundary conductance) and to a higher solar radiation intensity along the canopy profile. These differences did not affect fruit quality in terms of size, red skin over-colour and soluble solid content. In MrS 2/5, the higher light availability at all levels along canopy profile was related to a moderate water deficit status, even under full-irrigation conditions, as evidenced by the lower stem water potential (below -1.3 MPa) and by a lower transpiration rate (about one-half of the values observed on GF 677). At tree-level, MrS 2/5 had a daily water consumption that, also in relation to the lower leaf area per tree, resulted as low as 25% of the values observed on GF 677. The latter, even carrying a significantly higher leaf area and higher water consumption, never showed apparent symptoms of water deficit [it

  1. Loss of a highly conserved sterile alpha motif domain gene (WEEP) results in pendulous branch growth in peach trees.

    Science.gov (United States)

    Hollender, Courtney A; Pascal, Thierry; Tabb, Amy; Hadiarto, Toto; Srinivasan, Chinnathambi; Wang, Wanpeng; Liu, Zhongchi; Scorza, Ralph; Dardick, Chris

    2018-05-15

    Plant shoots typically grow upward in opposition to the pull of gravity. However, exceptions exist throughout the plant kingdom. Most conspicuous are trees with weeping or pendulous branches. While such trees have long been cultivated and appreciated for their ornamental value, the molecular basis behind the weeping habit is not known. Here, we characterized a weeping tree phenotype in Prunus persica (peach) and identified the underlying genetic mutation using a genomic sequencing approach. Weeping peach tree shoots exhibited a downward elliptical growth pattern and did not exhibit an upward bending in response to 90° reorientation. The causative allele was found to be an uncharacterized gene, Ppa013325 , having a 1.8-Kb deletion spanning the 5' end. This gene, dubbed WEEP , was predominantly expressed in phloem tissues and encodes a highly conserved 129-amino acid protein containing a sterile alpha motif (SAM) domain. Silencing WEEP in the related tree species Prunus domestica (plum) resulted in more outward, downward, and wandering shoot orientations compared to standard trees, supporting a role for WEEP in directing lateral shoot growth in trees. This previously unknown regulator of branch orientation, which may also be a regulator of gravity perception or response, provides insights into our understanding of how tree branches grow in opposition to gravity and could serve as a critical target for manipulating tree architecture for improved tree shape in agricultural and horticulture applications. Copyright © 2018 the Author(s). Published by PNAS.

  2. Breeding rootstocks for Prunus species: Advances in genetic and genomics of peach and cherry as a model

    Directory of Open Access Journals (Sweden)

    Verónica Guajardo

    2015-08-01

    Full Text Available Prunus rootstock is an important choice in optimizing productivity of grafted cultivars. Nevertheless, many Prunus rootstocks are notoriously intolerant to hypoxia which is caused by waterlogging and/or heavy soils. There is no available information to help select Prunus rootstocks that are tolerant to stress conditions such as root hypoxia caused by excess moisture. Information from genetic maps has demonstrated a high level of synteny among Prunus species, and this suggests that they all share a similar genomic structure. It should be possible to identify the genetic determinants involved in tolerance to hypoxia and other traits in Prunus rootstocks by applying methods to identify regions of the genome involved in the expression of important traits; these have been developed mainly in peach which is the model species for the genus. Molecular markers that are tightly linked to major genes would be useful in marker-assisted selection (MAS to optimize new rootstock selection. This article provides insight on the advances in the development of molecular markers, genetic maps, and gene identification in Prunus, mainly in peach; the aim is to provide a gen