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Sample records for autophagosome-like vacuoles formation

  1. Ergosteryl-β-glucosidase (Egh1) involved in sterylglucoside catabolism and vacuole formation in Saccharomyces cerevisiae.

    Science.gov (United States)

    Watanabe, Takashi; Tani, Motohiro; Ishibashi, Yohei; Endo, Ikumi; Okino, Nozomu; Ito, Makoto

    2015-10-01

    Sterylglucosides (SGs) are composed of a glucose and sterol derivatives, and are distributed in fungi, plants and mammals. We recently identified EGCrP1 and EGCrP2 (endoglycoceramidase-related proteins 1 and 2) as a β-glucocerebrosidase and steryl-β-glucosidase, respectively, in Cryptococcus neoformans. We herein describe an EGCrP2 homologue (Egh1; ORF name, Yir007w) involved in SG catabolism in Saccharomyces cerevisiae. The purified recombinant Egh1 hydrolyzed various β-glucosides including ergosteryl β-glucoside (EG), cholesteryl β-glucoside, sitosteryl β-glucoside, para-nitrophenyl β-glucoside, 4-methylumberifellyl β-glucoside and glucosylceramide. The disruption of EGH1 in S. cerevisiae BY4741 (egh1Δ) resulted in the accumulation of EG and fragmentation of vacuoles. The expression of EGH1 in egh1Δ (revertant) reduced the accumulation of EG, and restored the morphology of vacuoles. The accumulation of EG was not detected in EGH1 and UGT51(ATG26) double-disrupted mutants (ugt51Δegh1Δ), indicating that EG was synthesized by Ugt51(Atg26) and degraded by Egh1 in vivo. These results clearly demonstrated that Egh1 is an ergosteryl-β-glucosidase that is functionally involved in the EG catabolic pathway and vacuole formation in S. cerevisiae.

  2. Trypanosoma cruzi: Entry Into Mammalian Host Cells and Parasitophorous Vacuole Formation

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    Emile Santos Barrias

    2013-08-01

    Full Text Available Trypanosoma cruzi, the causative agent of Chagas disease, is transmitted to vertebrate hosts by blood-sucking insects. This protozoan is an obligate intracellular parasite. The infective forms of the parasite are the metacyclic trypomastigotes, amastigotes and bloodstream trypomastigotes. The recognition between the parasite and mammalian host cell, involves numerous molecules present in both cell types, and similar to several intracellular pathogens, T.cruzi is internalized by host cells via multiple endocytic pathways. Morphological studies demonstrated that after the interaction of the infective forms of T.cruzi with phagocytic or non-phagocytic cell types, plasma membrane protrusions can form, showing similarity with those observed during canonical phagocytosis or macropinocytic events. Additionally, several molecules known to be molecular markers of membrane rafts, macropinocytosis and phagocytosis have been demonstrated to be present at the invasion site. These events may or may not depend on the host cell lysosomes and cytoskeleton. In addition, after penetration, components of the host endosomal-lysosomal system, such as early endosomes, late endosomes and lysosomes, participate in the formation of the nascent parasithophorous vacuole (VP. Dynamin, a molecule involved in vesicle formation, has been shown to be involved in the parasitophorous vacuole release from the host cell plasma membrane. This review focuses on the multiple pathways that T.cruzi can use to enter the host cells until complete VP formation. We will describe different endocytic processes, such as phagocytosis, macropinocytosis, endocytosis using membrane microdomains and clathrin-dependent endocytosis and show results that are consistent with their use by this smart parasite. We will also discuss other mechanisms that have been described, such as active penetration and the process that takes advantage of cell membrane wound repair.

  3. Cell-to-cell spread and massive vacuole formation after Cryptococcus neoformans infection of murine macrophages

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    Casadevall Arturo

    2007-08-01

    Full Text Available Abstract Background The interaction between macrophages and Cryptococcus neoformans (Cn is critical for containing dissemination of this pathogenic yeast. However, Cn can either lyse macrophages or escape from within them through a process known as phagosomal extrusion. Both events result in live extracellular yeasts capable of reproducing and disseminating in the extracellular milieu. Another method of exiting the intracellular confines of cells is through host cell-to-cell transfer of the pathogen, and this commonly occurs with the human immuno-deficiency virus (HIV and CD4+ T cells and macrophages. In this report we have used time-lapse imaging to determine if this occurs with Cn. Results Live imaging of Cryptococcus neoformans interactions with murine macrophages revealed cell-to-cell spread of yeast cells from infected donor cells to uninfected cells. Although this phenomenon was relatively rare its occurrence documents a new capacity for this pathogen to infect adjacent cells without exiting the intracellular space. Cell-to-cell spread appeared to be an actin-dependent process. In addition, we noted that cryptococcal phagosomal extrusion was followed by the formation of massive vacuoles suggesting that intracellular residence is accompanied by long lasting damage to host cells. Conclusion C. neoformans can escape the intracellular confines of macrophages in an actin dependent manner by cell-to-cell transfer of the yeast leading to infection of adjacent cells. In addition, complete extrusion of internalized Cn cells can lead to the formation of a massive vacuole which may be a sign of damage to the host macrophage. These observations document new outcomes for the interaction of C. neoformans with host cells that provide precedents for cell biological effects that may contribute to the pathogenesis of cryptococcal infections.

  4. The Vtc proteins in vacuole fusion: coupling NSF activity to V(0) trans-complex formation

    DEFF Research Database (Denmark)

    Müller, Oliver; Bayer, Martin J; Peters, Christopher;

    2002-01-01

    The fusion of cellular membranes comprises several steps; membrane attachment requires priming of SNAREs and tethering factors by Sec18p/NSF (N-ethylmaleimide sensitive factor) and LMA1. This leads to trans-SNARE pairing, i.e. formation of SNARE complexes between apposed membranes. The yeast...... vacuole system has revealed two subsequent molecular events: trans-complex formation of V-ATPase proteolipid sectors (V(0)) and release of LMA1 from the membrane. We have now identified a hetero-oligomeric membrane integral complex of vacuolar transporter chaperone (Vtc) proteins integrating these events....... The Vtc complex associates with the R-SNARE Nyv1p and with V(0). Subunits Vtc1p and Vtc4p control the initial steps of fusion. They are required for Sec18p/NSF activity in SNARE priming, membrane binding of LMA1 and V(0) trans-complex formation. In contrast, subunit Vtc3p is required for the latest step...

  5. The potential role of kinesin and dynein in Golgi scattering and cytoplasmic vacuole formation during acute experimental pancreatitis

    Institute of Scientific and Technical Information of China (English)

    Ina A Weber; Igor Buchwalow; Daniela Hahn; Wolfram Domschke; Markus M Lerch; Jürgen Schnekenburger

    2010-01-01

    @@ Dear Editor, Acute pancreatitis is initiated in pancreatic acinar cells and characterized by a profound disturbance in intracel-lular vesicle transport. Moreover, secretion of pancreatic zymogens is blocked, paralleled by the formation of cytoplasmic vacuoles with premature protease activation that precedes cellular necrosis and organ damage.

  6. Trypanosoma cruzi: Entry into Mammalian Host Cells and Parasitophorous Vacuole Formation

    Science.gov (United States)

    Barrias, Emile Santos; de Carvalho, Tecia Maria Ulisses; De Souza, Wanderley

    2013-01-01

    Trypanosoma cruzi, the causative agent of Chagas disease, is transmitted to vertebrate hosts by blood-sucking insects. This protozoan is an obligate intracellular parasite. The infective forms of the parasite are the metacyclic trypomastigotes, amastigotes, and bloodstream trypomastigotes. The recognition between the parasite and mammalian host cell, involves numerous molecules present in both cell types, and similar to several intracellular pathogens, T. cruzi is internalized by host cells via multiple endocytic pathways. Morphological studies demonstrated that after the interaction of the infective forms of T. cruzi with phagocytic or non-phagocytic cell types, plasma membrane (PM) protrusions can form, showing similarity with those observed during canonical phagocytosis or macropinocytic events. Additionally, several molecules known to be molecular markers of membrane rafts, macropinocytosis, and phagocytosis have been demonstrated to be present at the invasion site. These events may or may not depend on the host cell lysosomes and cytoskeleton. In addition, after penetration, components of the host endosomal-lysosomal system, such as early endosomes, late endosomes, and lysosomes, participate in the formation of the nascent parasitophorous vacuole (PV). Dynamin, a molecule involved in vesicle formation, has been shown to be involved in the PV release from the host cell PM. This review focuses on the multiple pathways that T. cruzi can use to enter the host cells until complete PV formation. We will describe different endocytic processes, such as phagocytosis, macropinocytosis, and endocytosis using membrane microdomains and clathrin-dependent endocytosis and show results that are consistent with their use by this smart parasite. We will also discuss others mechanisms that have been described, such as active penetration and the process that takes advantage of cell membrane wound repair. PMID:23914186

  7. Shape, shell, and vacuole formation during the drying of a single concentrated whey protein droplet.

    Science.gov (United States)

    Sadek, Céline; Tabuteau, Hervé; Schuck, Pierre; Fallourd, Yannick; Pradeau, Nicolas; Le Floch-Fouéré, Cécile; Jeantet, Romain

    2013-12-17

    The drying of milk concentrate droplets usually leads to specific particle morphology influencing their properties and their functionality. Understanding how the final shape of the particle is formed therefore represents a key issue for industrial applications. In this study, a new approach to the investigation of droplet-particle conversion is proposed. A single droplet of concentrated globular proteins extracted from milk was deposited onto a hydrophobic substrate and placed in a dry environment. Complementary methods (high-speed camera, confocal microscopy, and microbalance) were used to record the drying behavior of the concentrated protein droplets. Our results showed that whatever the initial concentration, particle formation included three dynamic stages clearly defined by the loss of mass and the evolution of the internal and external shapes of the droplet. A new and reproducible particle shape was related in this study. It was observed after drying a smooth, hemispherical cap-shaped particle, including a uniform protein shell and the nucleation of an internal vacuole. The particle morphology was strongly influenced by the drying environment, the contact angle, and the initial protein concentration, all of which governed the duration of the droplet shrinkage, the degree of buckling, and the shell thickness. These results are discussed in terms of specific protein behaviors in forming a predictable and a characteristic particle shape. The way the shell is formed may be the starting point in shaping particle distortion and thus represents a potential means of tuning the particle morphology.

  8. Ubiquilin/Dsk2 promotes inclusion body formation and vacuole (lysosome)-mediated disposal of mutated huntingtin.

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    Chuang, Kun-Han; Liang, Fengshan; Higgins, Ryan; Wang, Yanchang

    2016-07-01

    Ubiquilin proteins contain a ubiquitin-like domain (UBL) and ubiquitin-associated domain(s) that interact with the proteasome and ubiquitinated substrates, respectively. Previous work established the link between ubiquilin mutations and neurodegenerative diseases, but the function of ubiquilin proteins remains elusive. Here we used a misfolded huntingtin exon I containing a 103-polyglutamine expansion (Htt103QP) as a model substrate for the functional study of ubiquilin proteins. We found that yeast ubiquilin mutant (dsk2Δ) is sensitive to Htt103QP overexpression and has a defect in the formation of Htt103QP inclusion bodies. Our evidence further suggests that the UBL domain of Dsk2 is critical for inclusion body formation. Of interest, Dsk2 is dispensable for Htt103QP degradation when Htt103QP is induced for a short time before noticeable inclusion body formation. However, when the inclusion body forms after a long Htt103QP induction, Dsk2 is required for efficient Htt103QP clearance, as well as for autophagy-dependent delivery of Htt103QP into vacuoles (lysosomes). Therefore our data indicate that Dsk2 facilitates vacuole-mediated clearance of misfolded proteins by promoting inclusion body formation. Of importance, the defect of inclusion body formation in dsk2 mutants can be rescued by human ubiquilin 1 or 2, suggesting functional conservation of ubiquilin proteins.

  9. Formation of a pathogen vacuole according to Legionella pneumophila: how to kill one bird with many stones.

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    Finsel, Ivo; Hilbi, Hubert

    2015-07-01

    Legionella species are ubiquitous, waterborne bacteria that thrive in numerous ecological niches. Yet, in contrast to many other environmental bacteria, Legionella spp. are also able to grow intracellularly in predatory protozoa. This feature mainly accounts for the pathogenicity of Legionella pneumophila, which causes the majority of clinical cases of a severe pneumonia termed Legionnaires' disease. The pathomechanism underlying L. pneumophila infection is based on macrophage resistance, which in turn is largely defined by the opportunistic pathogen's resistance towards amoebae. L. pneumophila replicates in macrophages or amoebae in a unique membrane-bound compartment, the Legionella-containing vacuole (LCV). LCV formation requires the bacterial intracellular multiplication/defective for organelle trafficking (Icm/Dot) type IV secretion system and involves a plethora of translocated effector proteins, which subvert pivotal processes in the host cell. Of the ca. 300 different experimentally validated Icm/Dot substrates, about 50 have been studied and attributed a cellular function to date. The versatility and ingenuity of these effectors' mode of actions is striking. In this review, we summarize insight into the cellular functions and biochemical activities of well-characterized L. pneumophila effector proteins and the host pathways they target. Recent studies not only substantially increased our knowledge about pathogen-host interactions, but also shed light on novel biological mechanisms.

  10. Endocytic Trafficking towards the Vacuole Plays a Key Role in the Auxin Receptor SCFTIR-Independent Mechanism of Lateral Root Formation in A.thaliana

    Institute of Scientific and Technical Information of China (English)

    Patricio Pérez-Henríquez; Natasha V.Raikhel; Lorena Norambuena

    2012-01-01

    Plants' developmental plasticity plays a pivotal role in responding to environmental conditions.One of the most plastic plant organs is the root system.Different environmental stimuli such as nutrients and water deficiency may induce lateral root formation to compensate for a low level of water and/or nutrients.It has been shown that the hormone auxin tunes lateral root development and components for its signaling pathway have been identified.Using chemical biology,we discovered an Arabidopsis thaliana lateral root formation mechanism that is independent of the auxin receptor SCFTIR.The bioactive compound Sortin2 increased lateral root occurrence by acting upstream from the morphological marker of lateral root primordium formation,the mitotic activity.The compound did not display auxin activity.At the cellular level,Sortin2 accelerated endosomal trafficking,resulting in increased trafficking of plasma membrane recycling proteins to the vacuole.Sortin2 affected Late endosome/PVC/MVB trafficking and morphology.Combining Sortin2 with well-known drugs showed that endocytic trafficking of Late E/PVC/MVB towards the vacuole is pivotal for Sortin2induced SCFTIR-independent lateral root initiation.Our results revealed a distinctive role for endosomal trafficking in the promotion of lateral root formation via a process that does not rely on the auxin receptor complex SCFTIR.

  11. The neutral lipid composition present in the digestive vacuole of Plasmodium falciparum concentrates heme and mediates β-hematin formation with an unusually low activation energy.

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    Hoang, Anh N; Sandlin, Rebecca D; Omar, Aneesa; Egan, Timothy J; Wright, David W

    2010-11-30

    In eukaryotic cells, neutral lipids serve as major energy storage molecules; however, in Plasmodium falciparum, a parasite responsible for causing malaria in humans, neutral lipids may have other functions during the intraerythrocytic stage of the parasite life cycle. Specifically, experimental data suggest that neutral lipid structures behave as a catalyst for the crystallization of hemozoin, a detoxification byproduct of several blood-feeding organisms, including malaria parasites. Synthetic neutral lipid droplets (SNLDs) were produced by depositing a lipid blend solution comprised of mono- and diglycerides onto an aqueous surface. These lipid droplets are able to mediate the production of brown pigments that are morphologically and chemically identical to hemozoin. The partitioning of heme into these SNLDs was examined by employing Nile Red, a lipid specific dye. Soluble ferriprotoporphyrin IX was observed to spontaneously localize to the lipid droplets, partitioning in a pH-dependent manner with an estimated log P of 2.6. Interestingly, the pH profile of heme partitioning closely resembles that of β-hematin formation. Differential scanning calorimetry and kinetic studies demonstrated that the SNLDs provide a unique environment that promotes hemozoin formation. SNLD-mediated formation of the malaria pigment displayed an activation energy barrier lower than those of individual lipid components. In particular, lipid droplets composed of diglycerides displayed activation barriers lower than those composed of monoglycerides. This difference was attributed to the greater fluidity of these lipids. In conjunction with the known pattern of lipid body proliferation, it is suggested that neutral lipid structures within the digestive vacuole not only are the location of in vivo hemozoin formation but are also essential for the survival of the parasite by functioning as a kinetically competent and site specific mediator for heme detoxification.

  12. Formation of the food vacuole in Plasmodium falciparum: a potential role for the 19 kDa fragment of merozoite surface protein 1 (MSP1(19.

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    Anton R Dluzewski

    Full Text Available Plasmodium falciparum Merozoite Surface Protein 1 (MSP1 is synthesized during schizogony as a 195-kDa precursor that is processed into four fragments on the parasite surface. Following a second proteolytic cleavage during merozoite invasion of the red blood cell, most of the protein is shed from the surface except for the C-terminal 19-kDa fragment (MSP1(19, which is still attached to the merozoite via its GPI-anchor. We have examined the fate of MSP1(19 during the parasite's subsequent intracellular development using immunochemical analysis of metabolically labeled MSP1(19, fluorescence imaging, and immuno-electronmicroscopy. Our data show that MSP1(19 remains intact and persists to the end of the intracellular cycle. This protein is the first marker for the biogenesis of the food vacuole; it is rapidly endocytosed into small vacuoles in the ring stage, which coalesce to form the single food vacuole containing hemozoin, and persists into the discarded residual body. The food vacuole is marked by the presence of both MSP1(19 and the chloroquine resistance transporter (CRT as components of the vacuolar membrane. Newly synthesized MSP1 is excluded from the vacuole. This behavior indicates that MSP1(19 does not simply follow a classical lysosome-like clearance pathway, instead, it may play a significant role in the biogenesis and function of the food vacuole throughout the intra-erythrocytic phase.

  13. Formation of hydrogen sulfide from cysteine in Saccharomyces cerevisiae BY4742: genome wide screen reveals a central role of the vacuole.

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    Gal Winter

    Full Text Available Discoveries on the toxic effects of cysteine accumulation and, particularly, recent findings on the many physiological roles of one of the products of cysteine catabolism, hydrogen sulfide (H2S, are highlighting the importance of this amino acid and sulfur metabolism in a range of cellular activities. It is also highlighting how little we know about this critical part of cellular metabolism. In the work described here, a genome-wide screen using a deletion collection of Saccharomyces cerevisiae revealed a surprising set of genes associated with this process. In addition, the yeast vacuole, not previously associated with cysteine catabolism, emerged as an important compartment for cysteine degradation. Most prominent among the vacuole-related mutants were those involved in vacuole acidification; we identified each of the eight subunits of a vacuole acidification sub-complex (V1 of the yeast V-ATPase as essential for cysteine degradation. Other functions identified included translation, RNA processing, folate-derived one-carbon metabolism, and mitochondrial iron-sulfur homeostasis. This work identified for the first time cellular factors affecting the fundamental process of cysteine catabolism. Results obtained significantly contribute to the understanding of this process and may provide insight into the underlying cause of cysteine accumulation and H2S generation in eukaryotes.

  14. H1-antihistamines induce vacuolation in astrocytes through macroautophagy

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    Hu, Wei-Wei; Yang, Ying; Wang, Zhe; Shen, Zhe; Zhang, Xiang-Nan [Department of Pharmacology, Key Laboratory of Medical Neurobiology of the Ministry of Health of China, Zhejiang Province Key Laboratory of Neurobiology, School of Basic Medical Sciences, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, Zhejiang, 310058 (China); Wang, Guang-Hui [College of Pharmaceutical Sciences, Soochow University, Suzhou, 215123 (China); Chen, Zhong, E-mail: chenzhong@zju.edu.cn [Department of Pharmacology, Key Laboratory of Medical Neurobiology of the Ministry of Health of China, Zhejiang Province Key Laboratory of Neurobiology, School of Basic Medical Sciences, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, Zhejiang, 310058 (China)

    2012-04-15

    H1-antihistamines induce vacuolation in vascular smooth muscle cells, which may contribute to their cardiovascular toxicity. The CNS toxicity of H1-antihistamines may also be related to their non-receptor-mediated activity. The aim of this study was to investigate whether H1-antihistamines induce vacuolation in astrocytes and the mechanism involved. The H1-antihistamines induced large numbers of giant vacuoles in astrocytes. Such vacuoles were marked with both the lysosome marker Lysotracker Red and the alkalescent fluorescence dye monodansylcadaverine, which indicated that these vacuoles were lysosome-like acidic vesicles. Quantitative analysis of monodansylcadaverine fluorescence showed that the effect of H1-antihistamines on vacuolation in astrocytes was dose-dependent, and was alleviated by extracellular acidification, but aggravated by extracellular alkalization. The order of potency to induce vacuolation at high concentrations of H1-antihistamines (diphenhydramine > pyrilamine > astemizole > triprolidine) corresponded to their pKa ranking. Co-treatment with histamine and the histamine receptor-1 agonist trifluoromethyl toluidide did not inhibit the vacuolation. Bafilomycin A1, a vacuolar (V)-ATPase inhibitor, which inhibits intracellular vacuole or vesicle acidification, clearly reversed the vacuolation and intracellular accumulation of diphenhydramine. The macroautophagy inhibitor 3-methyladenine largely reversed the percentage of LC3-positive astrocytes induced by diphenhydramine, while only partly reversing the number of monodansylcadaverine-labeled vesicles. In Atg5{sup −/−} mouse embryonic fibroblasts, which cannot form autophagosomes, the number of vacuoles induced by diphenhydramine was less than that in wild-type cells. These results indicated that H1-antihistamines induce V-ATPase-dependent acidic vacuole formation in astrocytes, and this is partly mediated by macroautophagy. The pKa and alkalescent characteristic of H1-antihistamines may be the

  15. The role of suppression of p38 MAPK in cellular vacuole formation%阻断p38丝裂原活化蛋白激酶在细胞空泡形成中的作用

    Institute of Scientific and Technical Information of China (English)

    张春燕; 冯春红; 敬健雄; 段春燕; 刘友平; 夏先明; 李洪; 代荣阳; 陈绍坤

    2014-01-01

    目的:探讨p38丝裂原活化蛋白激酶(p38MAPK)通路与细胞空泡形成的关系。方法应用茴香霉素、放线菌酮、p38MAPK抑制剂SB203580、JNK抑制剂SP600125处理HepG2、LM3、QBC939、Hela和A549细胞,光学显微镜和激光共聚焦显微镜观察细胞空泡化情况;Westernblot法检测p38MAPK等通路相关分子的表达水平;内质网红色荧光探针标记内质网,激光共聚焦显微镜观察内质网结构变化;溶酶体红色荧光探针标记溶酶体,激光共聚焦显微镜观察溶酶体荧光染色情况。结果(1)茴香霉素对HepG2细胞空泡有消除作用。(2)茴香霉素通过活化p38MAPK消除细胞空泡。(3)阻断p38MAPK诱导多种肿瘤细胞空泡形成。(4)阻断p38MAPK介导的空泡形成破坏内质网结构的整体性。(5)阻断p38MAPK介导的空泡形成具有可逆性。结论p38MAPK通路在调节细胞空泡形成中发挥了重要作用。%Objective To investigate the role of the p38 MAPK pathway in the formation of cytoplasmic vacuoles .Methods Af-ter treated with Anisomycin ,SB203580 or SP600125 ,images of HepG2 ,LM3 ,QBC939 ,Hela and A549 cells were recorded by light microscopy and taken at a magnification of 400 × .The effects of anisomycin ,SB203580 and SP600125 on the activity of p38 and JNK were measured by Western blot .LM3 and A549 cells were stained with the ER-tracker red and the lyso-tracker red and subjec-ted to confocal microscopy analysis .Results (1)Anisomycin could abolish cytoplasmic vacuolization of HepG2 cells .(2)p38 MAPK activation was responsible for anisomycin-induced cytoplasmic vacuolization abolishment .(3)p38 MAPK blocking initiated cytoplas-mic vacuoles formation in various cancer cell lines .(4)p38 MAPK blocking-induced cytoplasmic vacuoles disrupted the integrity of endoplasmic reticulum .(5)p38 MAPK blocking reversibly induced cytoplasmic vacuoles formation .Conclusion These observations provide direct evidence for a

  16. Vacuolation induced by unfavorable pH in cyanobacteria

    Institute of Scientific and Technical Information of China (English)

    赵以军; 吴红艳; 郭厚良; 许敏; 程凯; 祝海燕

    2001-01-01

    Six species or strains of cyanobacteria, Anabaena sp. 595, Plectonema boryanum 246, Scytonema hofmanni 248, Nostoc sp. 96, Oscillatoria animlis 284 and Spirulina maxima 438, were cultured in unfavorable pH conditions for vacuole induction. At pH 5.0, 6.5, or 7.0, vacuoles were observed to form in both Anabaena sp. 595 and Plectonema boryanum 246, especially in the former. The vacuolation took place with some morphological changes, such as the cells being inflated, spherical and vacuolated, and with unequalized division. The induced vacuoles in An- abaena sp. 595 and Plectonema boryanum 246 were in spherical shape and in rather transparent appearance under a phase microscope. For Scytonema hofmanni 248, it was less sensitive to pH, its vacuole formation was found only at pH 6.5. No vacuolization occurred in the cells of Nostoc sp. 96, Oscillatoria animlis 284 and Spirulina maxima 438 at all pH conditions we used. Vacuolization under unfavorable pH provides a new proof for the existence of vacuole in cells of cyanobacteria and reflects the prokaryote's function in ecological environment.

  17. Cell vacuolation induced by Haemophilus influenzae supernatants in HEp-2 cells

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    Maria del Rosario Espinoza-Mellado

    2013-12-01

    Full Text Available Haemophilus influenzae belongs to respiratory tract microbiota. We observed vacuoles formation in previous studies with H. influenzae culture supernatants, so in this work we characterised that cytotoxic effect. We observed an abundant production of acidic cytoplasmic vacuoles due to the presence of a “vacuolating factor” in H. influenzae supernatants which was characterised as thermolabile. Greatest vacuolating activity was observed when utilizing the fraction > 50 kDa. The presence of a large number of vacuoles in HEp-2 cells was verified by transmission electron microscopy and some vacuoles were identified with a double membrane and/or being surrounded by ribosomes. These results suggest similar behaviour to that of vacuolating effects described by autotransporter proteins an undescribed cytotoxic effect induced by H. influenzae .

  18. Protein dynamics and proteolysis in plant vacuoles.

    Science.gov (United States)

    Müntz, Klaus

    2007-01-01

    Plant cells cannot live without their vacuoles. The tissues and organs of a plant contain a wide variety of differentiated and specialized vacuoles -- even a single plant cell can possess two or more types of vacuoles. Vacuolar proteins are encoded by nuclear genes and synthesized in the cytoplasm. Their transport into the vacuolar compartment is under cytoplasmic control. Transcription seems to be a major control level for differential protein supply to the vacuoles. It is at this level that vacuole differentiation and functions are mainly integrated into cellular processes. Recycling amino acids generated by protein degradation is a major function of the vacuole. This is most evident when storage proteins are mobilized in storage tissues of generative or vegetative organs in order to nourish the embryo of germinating seeds or sprouting buds. When specific proteins are transferred to the vacuole for immediate degradation this compartment contributes to the adaptation of protein complexes in response to changes in developmental or environmental conditions. Vacuolar proteases are involved in protein degradation during reversible senescence and programmed cell death, which is also called irreversible senescence. Vacuoles contribute to defence against pathogens and herbivores by limited and unlimited proteolysis. Our present knowledge on functions and processes of vacuolar protein dynamics in plants is reviewed. Research perspectives are deduced.

  19. Calcium Signals from the Vacuole

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    Gerald Schönknecht

    2013-10-01

    Full Text Available The vacuole is by far the largest intracellular Ca2+ store in most plant cells. Here, the current knowledge about the molecular mechanisms of vacuolar Ca2+ release and Ca2+ uptake is summarized, and how different vacuolar Ca2+ channels and Ca2+ pumps may contribute to Ca2+ signaling in plant cells is discussed. To provide a phylogenetic perspective, the distribution of potential vacuolar Ca2+ transporters is compared for different clades of photosynthetic eukaryotes. There are several candidates for vacuolar Ca2+ channels that could elicit cytosolic [Ca2+] transients. Typical second messengers, such as InsP3 and cADPR, seem to trigger vacuolar Ca2+ release, but the molecular mechanism of this Ca2+ release still awaits elucidation. Some vacuolar Ca2+ channels have been identified on a molecular level, the voltage-dependent SV/TPC1 channel, and recently two cyclic-nucleotide-gated cation channels. However, their function in Ca2+ signaling still has to be demonstrated. Ca2+ pumps in addition to establishing long-term Ca2+ homeostasis can shape cytosolic [Ca2+] transients by limiting their amplitude and duration, and may thus affect Ca2+ signaling.

  20. Cellular vacuoles induced by Mycoplasma pneumoniae CARDS toxin originate from Rab9-associated compartments.

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    Coreen Johnson

    Full Text Available Recently, we identified an ADP-ribosylating and vacuolating cytotoxin in Mycoplasma pneumoniae designated Community Acquired Respiratory Distress Syndrome (CARDS toxin. In this study we show that vacuoles induced by recombinant CARDS (rCARDS toxin are acidic and derive from the endocytic pathway as determined by the uptake of neutral red and the fluid-phase marker, Lucifer yellow, respectively. Also, we demonstrate that the formation of rCARDS toxin-associated cytoplasmic vacuoles is inhibited by the vacuolar ATPase inhibitor, bafilomycin A1, and the ionophore, monensin. To examine the ontogeny of these vacuoles, we analyzed the distribution of endosomal and lysosomal membrane markers during vacuole formation and observed the enrichment of the late endosomal GTPase, Rab9, around rCARDS toxin-induced vacuoles. Immunogold-labeled Rab9 and overexpression of green fluorescent-tagged Rab9 further confirmed vacuolar association. The late endosomal- and lysosomal-associated membrane proteins, LAMP1 and LAMP2, also localized to the vacuolar membranes, while the late endosomal protein, Rab7, and early endosomal markers, Rab5 and EEA1, were excluded. HeLa cells expressing dominant-negative (DN Rab9 exhibited markedly reduced vacuole formation in the presence of rCARDS toxin, in contrast to cells expressing DN-Rab7, highlighting the importance of Rab9 function in rCARDS toxin-induced vacuolation. Our findings reveal the unique Rab9-association with rCARDS toxin-induced vacuoles and its possible relationship to the characteristic histopathology that accompanies M. pneumoniae infection.

  1. Effect of the Vacuolation of Helicobacter Pylori

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Cytotoxic test in vitro combined with cytochemical stain, fluorescent stain, transmission electronmicrograph was used to study the vacuolated effect by helicobacter pylori (H.pylori) (Toxin+) and its pathological mechanism. 78.26 % patients with peptic ulcer associated with H.pylori was infected with H.pylori (Toxin+), while 42.86 % patients with gastritis was infected with H.pylori (Toxin+). It was positive in vacuole with acridine orange and acid phosphatase stain. Transmission electronmicrograph of vacuole revealed the presence of abounding membrane. There was a closed relationship between infection with H.pylori (Toxin+) and peptic ulcer disease. The vacuole induced by H.pylori (Toxin+) was autophagosome, which was pathological phenomenon induced by toxin.

  2. Leishmania amazonensis Engages CD36 to Drive Parasitophorous Vacuole Maturation.

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    Kendi Okuda

    2016-06-01

    Full Text Available Leishmania amastigotes manipulate the activity of macrophages to favor their own success. However, very little is known about the role of innate recognition and signaling triggered by amastigotes in this host-parasite interaction. In this work we developed a new infection model in adult Drosophila to take advantage of its superior genetic resources to identify novel host factors limiting Leishmania amazonensis infection. The model is based on the capacity of macrophage-like cells, plasmatocytes, to phagocytose and control the proliferation of parasites injected into adult flies. Using this model, we screened a collection of RNAi-expressing flies for anti-Leishmania defense factors. Notably, we found three CD36-like scavenger receptors that were important for defending against Leishmania infection. Mechanistic studies in mouse macrophages showed that CD36 accumulates specifically at sites where the parasite contacts the parasitophorous vacuole membrane. Furthermore, CD36-deficient macrophages were defective in the formation of the large parasitophorous vacuole typical of L. amazonensis infection, a phenotype caused by inefficient fusion with late endosomes and/or lysosomes. These data identify an unprecedented role for CD36 in the biogenesis of the parasitophorous vacuole and further highlight the utility of Drosophila as a model system for dissecting innate immune responses to infection.

  3. ER and vacuoles: never been closer

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    Corrado eViotti

    2014-02-01

    Full Text Available The endoplasmic reticulum (ER represents the gateway for intracellular trafficking of membrane proteins, soluble cargoes and lipids. In all eukaryotes, the best described mechanism of exiting the ER is via COPII-coated vesicles, which transport both membrane proteins and soluble cargo to the cis-Golgi. The vacuole, together with the plasma membrane, is the most distal point of the secretory pathway, and many vacuolar proteins are transported from the ER through intermediate compartments. However, past results and recent findings demonstrate the presence of alternative transport routes from the ER towards the tonoplast, which are independent of Golgi- and post-Golgi trafficking. Moreover, the transport mechanism of the vacuolar proton pumps VHA-a3 and AVP1 challenges the current model of vacuole biogenesis, pointing to the endoplasmic reticulum for being the main membrane source for the biogenesis of the plant lytic compartment. This review gives an overview of the current knowledge on the transport routes towards the vacuole and discusses the possible mechanism of vacuole biogenesis in plants.

  4. Purification and proteomics of pathogen-modified vacuoles and membranes

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    Jo-Ana eHerweg

    2015-06-01

    Full Text Available Certain pathogenic bacteria adopt an intracellular lifestyle and proliferate in eukaryotic host cells. The intracellular niche protects the bacteria from cellular and humoral components of the mammalian immune system, and at the same time, allows the bacteria to gain access to otherwise restricted nutrient sources. Yet, intracellular protection and access to nutrients comes with a price, i.e. the bacteria need to overcome cell-autonomous defense mechanisms, such as the bactericidal endocytic pathway. While a few bacteria rupture the early phagosome and escape into the host cytoplasm, most intracellular pathogens form a distinct, degradation-resistant and replication-permissive membranous compartment. Intracellular bacteria that form unique pathogen vacuoles include Legionella, Mycobacterium, Chlamydia, Simkania and Salmonella species. In order to understand the formation of these pathogen niches on a global scale and in a comprehensive and quantitative manner, an inventory of compartment-associated host factors is required. To this end, the intact pathogen compartments need to be isolated, purified and biochemically characterized. Here, we review recent progress on the isolation and purification of pathogen-modified vacuoles and membranes, as well as their proteomic characterization by mass spectrometry and different validation approaches. These studies provide the basis for further investigations on the specific mechanisms of pathogen-driven compartment formation.

  5. Helicobacter pylori vacuolating toxin A and apoptosis

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    Rassow Joachim

    2011-11-01

    Full Text Available Abstract VacA, the vacuolating cytotoxin A of Helicobacter pylori, induces apoptosis in epithelial cells of the gastic mucosa and in leukocytes. VacA is released by the bacteria as a protein of 88 kDa. At the outer surface of host cells, it binds to the sphingomyelin of lipid rafts. At least partially, binding to the cells is facilitated by different receptor proteins. VacA is internalized by a clathrin-independent mechanism and initially accumulates in GPI-anchored proteins-enriched early endosomal compartments. Together with early endosomes, VacA is distributed inside the cells. Most of the VacA is eventually contained in the membranes of vacuoles. VacA assembles in hexameric oligomers forming an anion channel of low conductivity with a preference for chloride ions. In parallel, a significant fraction of VacA can be transferred from endosomes to mitochondria in a process involving direct endosome-mitochondria juxtaposition. Inside the mitochondria, VacA accumulates in the mitochondrial inner membrane, probably forming similar chloride channels as observed in the vacuoles. Import into mitochondria is mediated by the hydrophobic N-terminus of VacA. Apoptosis is triggered by loss of the mitochondrial membrane potential, recruitment of Bax and Bak, and release of cytochrome c.

  6. Pathogen vacuole purification from legionella-infected amoeba and macrophages.

    Science.gov (United States)

    Hoffmann, Christine; Finsel, Ivo; Hilbi, Hubert

    2013-01-01

    Legionella pneumophila replicates intracellularly in environmental and immune phagocytes within a unique membrane-bound compartment, the Legionella-containing vacuole (LCV). Formation of LCVs is strictly dependent on the Icm/Dot type IV secretion system and the translocation of "effector" proteins into the cell. Some effector proteins decorate the LCV membrane and subvert host cell vesicle trafficking pathways. Here we describe a method to purify intact LCVs from Dictyostelium discoideum amoebae and RAW 264.7 murine macrophages. The method comprises a two-step protocol: first, LCVs are enriched by immuno-magnetic separation using an antibody against a bacterial effector protein specifically localizing to the LCV membrane, and second, the LCVs are further purified by density gradient centrifugation. The purified LCVs can be characterized by proteomics and other biochemical approaches.

  7. Pleiotropic actions of Helicobacter pylori vacuolating cytotoxin, VacA.

    Science.gov (United States)

    Isomoto, Hajime; Moss, Joel; Hirayama, Toshiya

    2010-01-01

    Helicobacter pylori produces a vacuolating cytotoxin, VacA, and most virulent H. pylori strains secrete VacA. VacA binds to two types of receptor-like protein tyrosine phosphatase (RPTP), RPTPalpha and RPTPbeta, on the surface of host cells. VacA bound to RPTPbeta, relocates and concentrates in lipid rafts in the plasma membrane. VacA causes vacuolization, membrane anion-selective channel and pore formation, and disruption of endosomal and lysosomal activity in host cells. Secreted VacA is processed into p33 and p55 fragments. The p55 domain not only plays a role in binding to target cells but also in the formation of oligomeric structures and anionic membrane channels. Oral administration of VacA to wild-type mice, but not to RPTPbeta knockout mice, resulted in gastric ulcers, in agreement with the clinical effect of VacA. VacA with s1/m1 allele has more potent cytotoxic activity in relation to peptic ulcer disease and appears to be associated with human gastric cancer. VacA activates pro-apoptotic Bcl-2 family proteins, and induces apoptosis via a mitochondria-dependent pathway. VacA can disrupt other signal transduction pathways; VacA activates p38 MAPK, enhancing production of IL-8 and PGE(2), and PI3K/Akt, suppressing GSK-3beta activity. VacA has immunomodulatory actions on T cells and other immune cells, possibly contributing to the chronic infection seen with this organism. H. pylori virulence factors including VacA and CagA, which is encoded by cytotoxin-associated gene A, along with host genetic and environmental factors, constitute a complex network to regulate chronic gastric injury and inflammation, which is involved in a multistep process leading to gastric carcinogenesis.

  8. Role of endodermal cell vacuoles in shoot gravitropism.

    Science.gov (United States)

    Kato, Takehide; Morita, Miyo Terao; Tasaka, Masao

    2002-06-01

    In higher plants, shoots and roots show negative and positive gravitropism, respectively. Data from surgical ablation experiments and analysis of starch deficient mutants have led to the suggestion that columella cells in the root cap function as gravity perception cells. On the other hand, endodermal cells are believed to be the statocytes (that is, gravity perceiving cells) of shoots. Statocytes in shoots and roots commonly contain amyloplasts which sediment under gravity. Through genetic research with Arabidopsis shoot gravitropism mutants, sgr1/scr and sgr7/shr, it was determined that endodermal cells are essential for shoot gravitropism. Moreover, some starch biosynthesis genes and EAL1 are important for the formation and maturation of amyloplasts in shoot endodermis. Thus, amyloplasts in the shoot endodermis would function as statoliths, just as in roots. The study of the sgr2 and zig/sgr4 mutants provides new insights into the early steps of shoot gravitropism, which still remains unclear. SGR2 and ZIG/SGR4 genes encode a phospholipase-like and a v-SNARE protein, respectively. Moreover, these genes are involved in vacuolar formation or function. Thus, the vacuole must play an important role in amyloplast sedimentation because the sgr2 and zig/sgr4 mutants display abnormal amyloplast sedimentation.

  9. V-ATPase, ScNhxlp and Yeast Vacuole Fusion

    Institute of Scientific and Technical Information of China (English)

    Quan-Sheng Qiu

    2012-01-01

    Membrane fusion is the last step in trafficking pathways during which membrane vesicles fuse with target organelles to deliver cargos.It is a central cellular reaction that plays important roles in signal transduction,protein sorting and subcellular compartmentation.Recent progress in understanding the roles of ion transporters in vacuole fusion in yeast is summanzed in this article.It is becoming increasingly evident that the vacuolar proton pump V-ATPase and vacuolar Na+/H+ antiporter ScNhxlp are key components of the vacuole fusion machinery in yeast.Yeast ScNhxlp regulates vacuole fusion by controlling the luminal pH.V-ATPases serve a dual role in vacuolar integrity in which they regulate both vacuole fusion and fission reactions in yeast.Fission defects are epistatic to fusion defects.Vacuole fission depends on the proton translocation activity of the V-ATPase; by contrast,the fusion reaction does not need the transport activity but requires the physical presence of the proton pump.Vo,the membrane-integral sector of the V-ATPase,forms trans-complexes between the opposing vacuoles in the terminal phase of vacuole fusion where the Vo trans-complexes build a continuous proteolipid channel at the fusion site to mediate the bilayer fusion.

  10. Construction and identification of the eukaryotic expression vector for gene encoding the vacuolating cytotoxin of Helicobacter pylori and observations on the induction of macrophage vacuole formation and apoptosis%幽门螺杆菌VacA N端真核表达载体的构建、鉴定及其诱导巨噬细胞空泡化和凋亡的观察

    Institute of Scientific and Technical Information of China (English)

    黎村艳; 张艳; 刘志杰; 余敏君; 于文

    2008-01-01

    目的 构建幽门螺杆菌(Helicobacter pylori,HP)空泡毒素(Vacuolating cytotoxin,VacA)基因的真核表达载体pDsRed-Monomer-C1/vacA,并在THP-1巨噬细胞中表达,为研究VacA单一毒力决定簇的致病性奠定实验基础.方法 用Primer 5.0软件设计引物,以HP基因组为模板,PCR扩增vacA目的基因片段,克隆入真核表达载体pDsRed-Monomer-C1中,经酶切、PCR鉴定及测序鉴定后,转染 THP-1巨噬细胞中,荧光显微镜和Western-blot检测VacA蛋白在细胞中的表达;电镜观察巨噬细胞(MΦ)的空泡样变和凋亡;流式细胞仪检测细胞的凋亡率.结果 PCR扩增得到了大小约为1 428bp的目的片段,双酶切及测序鉴定证明成功构建了HP真核表达重组载体;转染后24h,重组质粒组部分细胞中有聚集的荧光颗粒,部分细胞发生空泡样变和凋亡改变;细胞凋亡率明显高于空质粒组和阴性对照组(P<0.001),细胞核因子-κB(nuclear factor kappaB,NF-κB)的抑制剂二硫代氨基甲酸吡咯烷(pyrrolidine dithiocarbamate,PDTC)抑制细胞的凋亡.结论 VacA蛋白瞬时高表达促进THP-1巨噬细胞空泡样变和凋亡.NF-κB可能参与调节VacA诱导的巨噬细胞凋亡.

  11. Vacúolos de gás e flutuação em Difflugia mitriformis Wallich (Protista, Rhizopoda, Testaceolobosea Gas vacuoles and flotation in Diffugia mitriformis Wallich (Protista, Rhizopoda, Testaceolobosea

    Directory of Open Access Journals (Sweden)

    Vladimir Stolzenberg Torres

    1996-01-01

    Full Text Available The natural formation of gas vacuoles as a method of locomotion is described for Difflugia mitriformis Wallich, 1984. These vacuoles may contain different compositions of gases, basicly carbodioxyde or oxigen, with a membranous limitation similar or identical to other types of vacuoles. Those vacuoles are utilised by the organism as a mode of dislocation frorn the bottom to the water surface by flotation permiting better conditions for the survival of the individual, with the consequence of the perpetuance of the taxon.

  12. Molecular markers for granulovacuolar degeneration are present in rimmed vacuoles.

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    Masahiro Nakamori

    Full Text Available BACKGROUND: Rimmed vacuoles (RVs are round-oval cytoplasmic inclusions, detected in muscle cells of patients with myopathies, such as inclusion body myositis (IBM and distal myopathy with RVs (DMRV. Granulovacuolar degeneration (GVD bodies are spherical vacuoles containing argentophilic and hematoxyphilic granules, and are one of the pathological hallmarks commonly found in hippocampal pyramidal neurons of patients with aging-related neurodegenerative diseases, such as Alzheimer's disease and Parkinson's disease. These diseases are common in the elderly and share some pathological features. Therefore, we hypothesized that mechanisms of vacuolar formation in RVs and GVD bodies are common despite their role in two differing pathologies. We explored the components of RVs by immunohistochemistry, using antibodies for GVD markers. METHODS: Subjects included one AD case, eight cases of sporadic IBM, and three cases of DMRV. We compared immunoreactivity and staining patterns for GVD markers. These markers included: (1 tau-modifying proteins (caspase 3, cyclin-dependent kinase 5 [CDK5], casein kinase 1δ [CK1δ], and c-jun N-terminal kinase [JNK], (2 lipid raft-associated materials (annexin 2, leucine-rich repeat kinase 2 [LRRK2], and flotillin-1, and (3 other markers (charged multi-vesicular body protein 2B [CHMP2B] and phosphorylated transactive response DNA binding protein-43 [pTDP43] in both GVD bodies and RVs. Furthermore, we performed double staining of each GVD marker with pTDP43 to verify the co-localization. RESULTS: GVD markers, including lipid raft-associated proteins and tau kinases, were detected in RVs. CHMP2B, pTDP43, caspase 3, LRRK2, annexin 2 and flotillin-1 were detected on the rim and were diffusely distributed in the cytoplasm of RV-positive fibers. CDK5, CK1δ and JNK were detected only on the rim. In double staining experiments, all GVD markers colocalized with pTDP43 in RVs. CONCLUSIONS: These results suggest that RVs of muscle

  13. LegC3, an effector protein from Legionella pneumophila, inhibits homotypic yeast vacuole fusion in vivo and in vitro.

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    Terry L Bennett

    Full Text Available During infection, the intracellular pathogenic bacterium Legionella pneumophila causes an extensive remodeling of host membrane trafficking pathways, both in the construction of a replication-competent vacuole comprised of ER-derived vesicles and plasma membrane components, and in the inhibition of normal phagosome:endosome/lysosome fusion pathways. Here, we identify the LegC3 secreted effector protein from L. pneumophila as able to inhibit a SNARE- and Rab GTPase-dependent membrane fusion pathway in vitro, the homotypic fusion of yeast vacuoles (lysosomes. This vacuole fusion inhibition appeared to be specific, as similar secreted coiled-coiled domain containing proteins from L. pneumophila, LegC7/YlfA and LegC2/YlfB, did not inhibit vacuole fusion. The LegC3-mediated fusion inhibition was reversible by a yeast cytosolic extract, as well as by a purified soluble SNARE, Vam7p. LegC3 blocked the formation of trans-SNARE complexes during vacuole fusion, although we did not detect a direct interaction of LegC3 with the vacuolar SNARE protein complexes required for fusion. Additionally, LegC3 was incapable of inhibiting a defined synthetic model of vacuolar SNARE-driven membrane fusion, further suggesting that LegC3 does not directly inhibit the activity of vacuolar SNAREs, HOPS complex, or Sec17p/18p during membrane fusion. LegC3 is likely utilized by Legionella to modulate eukaryotic membrane fusion events during pathogenesis.

  14. Rimmed vacuoles with beta-amyloid and ubiquitinated filamentous deposits in the muscles of patients with long-standing denervation (postpoliomyelitis muscular atrophy): similarities with inclusion body myositis.

    Science.gov (United States)

    Semino-Mora, C; Dalakas, M C

    1998-10-01

    In the chronically denervated muscles of patients with prior paralytic poliomyelitis, there are secondary myopathic features, including endomysial inflammation and rare vacuolated fibers. To assess the frequency and characteristics of the vacuoles and their similarities with those seen in inclusion body myositis (IBM), we examined 58 muscle biopsy specimens from patients with prior paralytic poliomyelitis for (1) the presence of rimmed vacuoles; (2) acid-phosphatase reactivity; (3) Congo-red-positive amyloid deposits; (4) electron microscopy, searching for tubulofilaments; and (5) immunoelectron microscopy, using antibodies against beta-amyloid and ubiquitin. We found vacuolated muscle fibers in 18 of 58 (31%) biopsies, with a mean frequency of 2.06 +/- 0.42 fibers per specimen. The vacuoles contained acid phosphatase-positive material in 6 of the 18 (33.30%) specimens and stained positive for Congo red in five (27.80%). By immunoelectron microscopy, the vacuoles contained 5.17 +/- 0.13 nm fibrils and 14.9 +/- 0.31 nm filaments that immunoreacted with antibodies to beta-amyloid and ubiquitin in a pattern identical to the one seen in IBM. We conclude that vacuolated muscle fibers containing filamentous inclusions positive for amyloid and ubiquitin are not unique to IBM and the other vacuolar myopathies but can also occur in a chronic neurogenic condition, such as postpoliomyelitis. The chronicity of the underlying disease, rather than the cause, may lead to vacuolar formation, amyloid deposition, and accumulation of ubiquitinated filaments.

  15. Effect of Helicobacter pylori's vacuolating cytotoxin on the autophagy pathway in gastric epithelial cells.

    Science.gov (United States)

    Terebiznik, Mauricio R; Raju, Deepa; Vázquez, Cristina L; Torbricki, Karl; Kulkarni, Reshma; Blanke, Steven R; Yoshimori, Tamotsu; Colombo, María I; Jones, Nicola L

    2009-04-01

    Host cell responses to Helicobacter pylori infection are complex and incompletely understood. Here, we report that autophagy is induced within human-derived gastric epithelial cells (AGS) in response to H. pylori infection. These autophagosomes were distinct and different from the large vacuoles induced during H. pylori infection. Autophagosomes were detected by transmission electron microscopy, conversion of LC3-I to LC3-II, GFP-LC3 recruitment to autophagosomes, and depended on Atg5 and Atg12. The induction of autophagy depended on the vacuolating cytotoxin (VacA) and, moreover, VacA was sufficient to induce autophagosome formation. The channel-forming activity of VacA was necessary for inducing autophagy. Intracellular VacA partially co-localized with GFP-LC3, indicating that the toxin associates with autophagosomes. The inhibition of autophagy increased the stability of intracellular VacA, which in turn resulted in enhanced toxin-mediated cellular vacuolation. These findings suggest that the induction of autophagy by VacA may represent a host mechanism to limit toxin-induced cellular damage.

  16. Interorganelle interactions and inheritance patterns of nuclei and vacuoles in budding yeast meiosis.

    Science.gov (United States)

    Tsai, I-Ting; Lin, Jyun-Liang; Chiang, Yi-Hsuan; Chuang, Yu-Chien; Liang, Shu-Shan; Chuang, Chi-Ning; Huang, Tzyy-Nan; Wang, Ting-Fang

    2014-02-01

    Many of the mechanisms by which organelles are inherited by spores during meiosis are not well understood. Dramatic chromosome motion and bouquet formation are evolutionarily conserved characteristics of meiotic chromosomes. The budding yeast bouquet genes (NDJ1, MPS3, CSM4) mediate these movements via telomere attachment to the nuclear envelope (NE). Here, we report that during meiosis the NE is in direct contact with vacuoles via nucleus-vacuole junctions (NVJs). We show that in meiosis NVJs are assembled through the interaction of the outer NE-protein Nvj1 and the vacuolar membrane protein Vac8. Notably, NVJs function as diffusion barriers that exclude the nuclear pore complexes, the bouquet protein Mps3 and NE-tethered telomeres from the outer nuclear membrane and nuclear ER, resulting in distorted NEs during early meiosis. An increase in NVJ area resulting from Nvj1-GFP overexpression produced a moderate bouquet mutant-like phenotype in wild-type cells. NVJs, as the vacuolar contact sites of the nucleus, were found to undergo scission alongside the NE during meiotic nuclear division. The zygotic NE and NVJs were partly segregated into 4 spores. Lastly, new NVJs were also revealed to be synthesized de novo to rejoin the zygotic NE with the newly synthesized vacuoles in the mature spores. In conclusion, our results revealed that budding yeast nuclei and vacuoles exhibit dynamic interorganelle interactions and different inheritance patterns in meiosis, and also suggested that nvj1Δ mutant cells may be useful to resolve the technical challenges pertaining to the isolation of intact nuclei for the biochemical study of meiotic nuclear proteins.

  17. Organelle acidification negatively regulates vacuole membrane fusion in vivo

    Science.gov (United States)

    Desfougères, Yann; Vavassori, Stefano; Rompf, Maria; Gerasimaite, Ruta; Mayer, Andreas

    2016-01-01

    The V-ATPase is a proton pump consisting of a membrane-integral V0 sector and a peripheral V1 sector, which carries the ATPase activity. In vitro studies of yeast vacuole fusion and evidence from worms, flies, zebrafish and mice suggested that V0 interacts with the SNARE machinery for membrane fusion, that it promotes the induction of hemifusion and that this activity requires physical presence of V0 rather than its proton pump activity. A recent in vivo study in yeast has challenged these interpretations, concluding that fusion required solely lumenal acidification but not the V0 sector itself. Here, we identify the reasons for this discrepancy and reconcile it. We find that acute pharmacological or physiological inhibition of V-ATPase pump activity de-acidifies the vacuole lumen in living yeast cells within minutes. Time-lapse microscopy revealed that de-acidification induces vacuole fusion rather than inhibiting it. Cells expressing mutated V0 subunits that maintain vacuolar acidity were blocked in this fusion. Thus, proton pump activity of the V-ATPase negatively regulates vacuole fusion in vivo. Vacuole fusion in vivo does, however, require physical presence of a fusion-competent V0 sector. PMID:27363625

  18. Vacuole-targeting fungicidal activity of amphotericin B

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    Akira eOgita

    2012-03-01

    Full Text Available Invasive fungal infections are recognized as major threats to patients with immune depression as well as those with cancer chemotherapy. Amphotericin B (AmB, a classical antifungal agent with a polyene macrolide structure, is widely used for the control of serious fungal infections. However, the clinical use of this antibiotic is limited by the treatment-associated side effects and the appearance of resistant strains. AmB lethality has been generally elucidated by the alteration of plasma membrane ion permeability due to its specific binding to plasma membrane ergosterol. While, the recent studies with Saccharomyces cerevisiae and Candida albicans reveals the vacuole disruptive action as another cause of AmB lethality on the basis of its marked amplification in combination with allicin, an allyl sulfur compound from garlic. Indeed, AmB causes a serious structural damage to the vacuole membrane at a lethal concentration, and even at a non-lethal concentration in combination with allicin. Such an enhancement effect of allicin is dependent on an inhibition of ergosterol-trafficking from the plasma membrane to the vacuole membrane, which is considered to be a cellular response to protect against the vacuole membrane disintegration. Allicin can also decrease the minimum fungicidal concentration of AmB against the pathogenic fungi C. albicans and Aspergillus fumigatus, as is the case of S. cerevisiae. The synergistic fungicidal activities of AmB and allicin may have significant implications in the development of the vacuole-targeting chemotherapy against fungal infections.

  19. PIKfyve Regulates Vacuole Maturation and Nutrient Recovery following Engulfment.

    Science.gov (United States)

    Krishna, Shefali; Palm, Wilhelm; Lee, Yongchan; Yang, Wendy; Bandyopadhyay, Urmi; Xu, Haoxing; Florey, Oliver; Thompson, Craig B; Overholtzer, Michael

    2016-09-12

    The scavenging of extracellular macromolecules by engulfment can sustain cell growth in a nutrient-depleted environment. Engulfed macromolecules are contained within vacuoles that are targeted for lysosome fusion to initiate degradation and nutrient export. We have shown that vacuoles containing engulfed material undergo mTORC1-dependent fission that redistributes degraded cargo back into the endosomal network. Here we identify the lipid kinase PIKfyve as a regulator of an alternative pathway that distributes engulfed contents in support of intracellular macromolecular synthesis during macropinocytosis, entosis, and phagocytosis. We find that PIKfyve regulates vacuole size in part through its downstream effector, the cationic transporter TRPML1. Furthermore, PIKfyve promotes recovery of nutrients from vacuoles, suggesting a potential link between PIKfyve activity and lysosomal nutrient export. During nutrient depletion, PIKfyve activity protects Ras-mutant cells from starvation-induced cell death and supports their proliferation. These data identify PIKfyve as a critical regulator of vacuole maturation and nutrient recovery during engulfment.

  20. Vps1 in the late endosome-to-vacuole traffic

    Indian Academy of Sciences (India)

    Jacob Hayden; Michelle Williams; Ann Granich; Hyoeun Ahn; Brandon Tenay; Joshua Lukehart; Chad Highfill; Sarah Dobard; Kyoungtae Kim

    2013-03-01

    Vacuolar protein sorting 1 (Vps1), the yeast homolog to human dynamin, is a GTP hydrolyzing protein, which plays an important role in protein sorting and targeting between the Golgi and late endosomal compartments. In this study, we assessed the functional significance of Vps1 in the membrane traffic towards the vacuole. We show here that vps1 cells accumulated FM4-64 to a greater extent than wild-type (WT) cells, suggesting slower endocytic degradation traffic toward the vacuole. In addition, we observed that two endosome-to-vacuole traffic markers, DsRed-FYVE and Ste2-GFP, were highly accumulated in Vps1-deficient cells, further supporting Vps1’s implication in efficient trafficking of endocytosed materials to the vacuole. Noteworthy, a simultaneous imaging analysis in conjunction with FM4-64 pulse-chase experiment further revealed that Vps1 plays a role in late endosome to the vacuole transport. Consistently, our subcellular localization analysis showed that Vps1 is present at the late endosome. The hyperaccumulation of endosomal intermediates in the vps1 mutant cells appears to be caused by the disruption of integrity of HOPS tethering complexes, manifested by mislocalization of Vps39 to the cytoplasm. Finally, we postulate that Vps1 functions together with the Endosomal Sorting Complex Required for Transport (ESCRT) complex at the late endosomal compartments, based on the observation that the double mutants, in which VPS1 along with singular ESCRT I, II and III genes have been disrupted, exhibited synthetic lethality. Together, we propose that Vps1 is required for correct and efficient trafficking from the late endosomal compartments to the vacuole.

  1. Rimmed vacuoles in Becker muscular dystrophy have similar features with inclusion myopathies.

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    Kazunari Momma

    Full Text Available Rimmed vacuoles in myofibers are thought to be due to the accumulation of autophagic vacuoles, and can be characteristic in certain myopathies with protein inclusions in myofibers. In this study, we performed a detailed clinical, molecular, and pathological characterization of Becker muscular dystrophy patients who have rimmed vacuoles in muscles. Among 65 Becker muscular dystrophy patients, we identified 12 patients who have rimmed vacuoles and 11 patients who have deletions in exons 45-48 in DMD gene. All patients having rimmed vacuoles showed milder clinical features compared to those without rimmed vacuoles. Interestingly, the rimmed vacuoles in Becker muscular dystrophy muscles seem to represent autophagic vacuoles and are also associated with polyubiquitinated protein aggregates. These findings support the notion that rimmed vacuoles can appear in Becker muscular dystrophy, and may be related to the chronic changes in muscle pathology induced by certain mutations in the DMD gene.

  2. Rimmed vacuoles in Becker muscular dystrophy have similar features with inclusion myopathies.

    Science.gov (United States)

    Momma, Kazunari; Noguchi, Satoru; Malicdan, May Christine V; Hayashi, Yukiko K; Minami, Narihiro; Kamakura, Keiko; Nonaka, Ikuya; Nishino, Ichizo

    2012-01-01

    Rimmed vacuoles in myofibers are thought to be due to the accumulation of autophagic vacuoles, and can be characteristic in certain myopathies with protein inclusions in myofibers. In this study, we performed a detailed clinical, molecular, and pathological characterization of Becker muscular dystrophy patients who have rimmed vacuoles in muscles. Among 65 Becker muscular dystrophy patients, we identified 12 patients who have rimmed vacuoles and 11 patients who have deletions in exons 45-48 in DMD gene. All patients having rimmed vacuoles showed milder clinical features compared to those without rimmed vacuoles. Interestingly, the rimmed vacuoles in Becker muscular dystrophy muscles seem to represent autophagic vacuoles and are also associated with polyubiquitinated protein aggregates. These findings support the notion that rimmed vacuoles can appear in Becker muscular dystrophy, and may be related to the chronic changes in muscle pathology induced by certain mutations in the DMD gene.

  3. Characteristics of weak base-induced vacuoles formed around individual acidic organelles.

    Science.gov (United States)

    Hiruma, Hiromi; Kawakami, Tadashi

    2011-01-01

    We have previously found that the weak base 4-aminopyridine induces Brownian motion of acidic organelles around which vacuoles are formed, causing organelle traffic disorder in neurons. Our present study investigated the characteristics of vacuoles induced by weak bases (NH(4)Cl, aminopyridines, and chloroquine) using mouse cells. Individual vacuoles included acidic organelles identified by fluorescent protein expression. Mitochondria and actin filaments were extruded outside the vacuoles, composing the vacuole rim. Staining with amine-reactive fluorescence showed no protein/amino acid content in vacuoles. Thus, serous vacuolar contents are probably partitioned by viscous cytosol, other organelles, and cytoskeletons, but not membrane. The weak base (chloroquine) was immunochemically detected in intravacuolar organelles, but not in vacuoles. Early vacuolization was reversible, but long-term vacuolization caused cell death. The vacuolization and cell death were blocked by the vacuolar H(+)-ATPase inhibitor and Cl--free medium. Staining with LysoTracker or LysoSensor indicated that intravacuolar organelles were strongly acidic and vacuoles were slightly acidic. This suggests that vacuolization is caused by accumulation of weak base and H(+) in acidic organelles, driven by vacuolar H(+)-ATPase associated with Cl(-) entering, and probably by subsequent extrusion of H(+) and water from organelles to the surrounding cytoplasm.

  4. Shigella subverts the host recycling compartment to rupture its vacuole.

    Science.gov (United States)

    Mellouk, Nora; Weiner, Allon; Aulner, Nathalie; Schmitt, Christine; Elbaum, Michael; Shorte, Spencer L; Danckaert, Anne; Enninga, Jost

    2014-10-08

    Shigella enters epithlial cells via internalization into a vacuole. Subsequent vacuolar membrane rupture allows bacterial escape into the cytosol for replication and cell-to-cell spread. Bacterial effectors such as IpgD, a PI(4,5)P2 phosphatase that generates PI(5)P and alters host actin, facilitate this internalization. Here, we identify host proteins involved in Shigella uptake and vacuolar membrane rupture by high-content siRNA screening and subsequently focus on Rab11, a constituent of the recycling compartment. Rab11-positive vesicles are recruited to the invasion site before vacuolar rupture, and Rab11 knockdown dramatically decreases vacuolar membrane rupture. Additionally, Rab11 recruitment is absent and vacuolar rupture is delayed in the ipgD mutant that does not dephosphorylate PI(4,5)P₂ into PI(5)P. Ultrastructural analyses of Rab11-positive vesicles further reveal that ipgD mutant-containing vacuoles become confined in actin structures that likely contribute to delayed vacular rupture. These findings provide insight into the underlying molecular mechanism of vacuole progression and rupture during Shigella invasion.

  5. The virulence protein SopD2 regulates membrane dynamics of Salmonella-containing vacuoles.

    Directory of Open Access Journals (Sweden)

    Nina Schroeder

    Full Text Available Salmonella enterica serovar Typhimurium is a Gram-negative bacterial pathogen causing gastroenteritis in humans and a systemic typhoid-like illness in mice. The capacity of Salmonella to cause diseases relies on the establishment of its intracellular replication niche, a membrane-bound compartment named the Salmonella-containing vacuole (SCV. This requires the translocation of bacterial effector proteins into the host cell by type three secretion systems. Among these effectors, SifA is required for the SCV stability, the formation of Salmonella-induced filaments (SIFs and plays an important role in the virulence of Salmonella. Here we show that the effector SopD2 is responsible for the SCV instability that triggers the cytoplasmic release of a sifA(- mutant. Deletion of sopD2 also rescued intra-macrophagic replication and increased virulence of sifA(- mutants in mice. Membrane tubular structures that extend from the SCV are the hallmark of Salmonella-infected cells. Until now, these unique structures have not been observed in the absence of SifA. The deletion of sopD2 in a sifA(- mutant strain re-established membrane trafficking from the SCV and led to the formation of new membrane tubular structures, the formation of which is dependent on other Salmonella effector(s. Taken together, our data demonstrate that SopD2 inhibits the vesicular transport and the formation of tubules that extend outward from the SCV and thereby contributes to the sifA(- associated phenotypes. These results also highlight the antagonistic roles played by SopD2 and SifA in the membrane dynamics of the vacuole, and the complex actions of SopD2, SifA, PipB2 and other unidentified effector(s in the biogenesis and maintenance of the Salmonella replicative niche.

  6. Identification and characterization of receptors for vacuolating activity of subtilase cytotoxin.

    Science.gov (United States)

    Yahiro, Kinnosuke; Morinaga, Naoko; Satoh, Mamoru; Matsuura, Gen; Tomonaga, Takeshi; Nomura, Fumio; Moss, Joel; Noda, Masatoshi

    2006-10-01

    Some shiga toxin-producing Escherichia coli secrete a novel AB5 cytotoxin, named subtilase cytotoxin (SubAB), which induces vacuole formation in addition to cytotoxicity in susceptible cells. By immunoprecipitation with SubAB from Vero cells, we discovered proteins of 100 kDa, 135 kDa and 155 kDa as potential candidates for its receptor. These proteins were N-glycosylated in their extracellular domains, a modification that was necessary for interaction with SubAB. Biotinylated receptors were partially purified by Datura stramonium agglutinin affinity chromatography and avidin-agarose and analysed by TOF mass spectroscopy. The peptide sequences of p135 were identical to beta1 integrin, and its identification was confirmed with anti-integrin beta1 antibody. The p155 protein was identified as alpha2 integrin using anti-integrin alpha2 antibody. In addition, treatment of Vero cells with beta1 integrin RNAi before exposure to SubAB prevented vacuolating activity. These results suggested that SubAB recognizes alpha2beta1 integrin as a functional receptor; this first interaction may be an important key step leading to the SubAB-induced morphological changes in Vero cells.

  7. Thioploca spp: filamentous sulfur bacteria with nitrate vacuoles

    DEFF Research Database (Denmark)

    Jørgensen, BB; Gallardo, VA

    1999-01-01

    communities of large Thioploca species live along the Pacific coast of South America and in other upwelling areas of high organic matter sedimentation with bottom waters poor in oxygen and rich in nitrate. Each cell of these thioplocas harbors a large liquid vacuole which is used as a storage for nitrate...... with a concentration of lip to 506 mM. The nitrate is used as an electron acceptor for sulfide oxidation and the bacteria may grow autotrophically or mixotrophically using acetate or other organic molecules as carbon source. The filaments stretch up into the overlying seawater, from which they take up nitrate...

  8. Beyond Rab GTPases Legionella activates the small GTPase Ran to promote microtubule polymerization, pathogen vacuole motility, and infection.

    Science.gov (United States)

    Hilbi, Hubert; Rothmeier, Eva; Hoffmann, Christine; Harrison, Christopher F

    2014-01-01

    Legionella spp. are amoebae-resistant environmental bacteria that replicate in free-living protozoa in a distinct compartment, the Legionella-containing vacuole (LCV). Upon transmission of Legionella pneumophila to the lung, the pathogens employ an evolutionarily conserved mechanism to grow in LCVs within alveolar macrophages, thus triggering a severe pneumonia termed Legionnaires' disease. LCV formation is a complex and robust process, which requires the bacterial Icm/Dot type IV secretion system and involves the amazing number of 300 different translocated effector proteins. LCVs interact with the host cell's endosomal and secretory vesicle trafficking pathway. Accordingly, in a proteomics approach as many as 12 small Rab GTPases implicated in endosomal and secretory vesicle trafficking were identified and validated as LCV components. Moreover, the small GTPase Ran and its effector protein RanBP1 have been found to decorate the pathogen vacuole. Ran regulates nucleo-cytoplasmic transport, spindle assembly, and cytokinesis, as well as the organization of non-centrosomal microtubules. In L. pneumophila-infected amoebae or macrophages, Ran and RanBP1 localize to LCVs, and the small GTPase is activated by the Icm/Dot substrate LegG1. Ran activation by LegG1 leads to microtubule stabilization and promotes intracellular pathogen vacuole motility and bacterial growth, as well as chemotaxis and migration of Legionella-infected cells.

  9. Mitochondrial Extrusion through the cytoplasmic vacuoles during cell death.

    Science.gov (United States)

    Nakajima, Akihito; Kurihara, Hidetake; Yagita, Hideo; Okumura, Ko; Nakano, Hiroyasu

    2008-08-29

    Under various conditions, noxious stimuli damage mitochondria, resulting in mitochondrial fragmentation; however, the mechanisms by which fragmented mitochondria are eliminated from the cells remain largely unknown. Here we show that cytoplasmic vacuoles originating from the plasma membrane engulfed fragmented mitochondria and subsequently extruded them into the extracellular spaces in undergoing acute tumor necrosis factor alpha-induced cell death in a caspase-dependent fashion. Notably, upon fusion of the membrane encapsulating mitochondria to the plasma membrane, naked mitochondria were released into the extracellular spaces in an exocytotic manner. Mitochondrial extrusion was specific to tumor necrosis factor alpha-induced cell death, because a genotoxic stress-inducing agent such as cisplatin did not elicit mitochondrial extrusion. Moreover, intact actin and tubulin cytoskeletons were required for mitochondrial extrusion as well as membrane blebbing. Furthermore, fragmented mitochondria were engulfed by cytoplasmic vacuoles and extruded from hepatocytes of mice injected with anti-Fas antibody, suggesting that mitochondrial extrusion can be observed in vivo under pathological conditions. Mitochondria are eliminated during erythrocyte maturation under physiological conditions, and anti-mitochondrial antibody is detected in some autoimmune diseases. Thus, elucidating the mechanism underlying mitochondrial extrusion will open a novel avenue leading to better understanding of various diseases caused by mitochondrial malfunction as well as mitochondrial biology.

  10. The role of Plasmodium falciparum food vacuole plasmepsins.

    Science.gov (United States)

    Liu, Jun; Gluzman, Ilya Y; Drew, Mark E; Goldberg, Daniel E

    2005-01-14

    Plasmepsins (PMs) are thought to have an important function in hemoglobin degradation in the malarial parasite Plasmodium falciparum and have generated interest as antimalarial drug targets. Four paralogous plasmepsins reside in the food vacuole of P. falciparum. Targeted gene disruption by double crossover homologous recombination has been employed to study food vacuole plasmepsin function in cultured parasites. Parasite clones with deletions in each of the individual PM I, PM II, and HAP genes as well as clones with a double PM IV/PM I disruption have been generated. All of these clones lack the corresponding PMs, are viable, and appear morphologically normal. PM II and PM IV/I disruptions have longer doubling times than the 3D7 parental line in rich RPMI medium. This appears to be because of a decreased level of productive progeny rather than an increased cell cycle time. In amino acid-limited medium, all four knockouts exhibit slower growth than the parental strain. Compared with 3D7, knock-out clone sensitivity to aspartic and cysteine protease inhibitors is changed minimally. These results suggest substantial functional redundancy and have important implications for the design of antimalarial drugs. The slow growth phenotype may explain why P. falciparum has maintained four plasmepsin genes with overlapping functions.

  11. Enzymic and protein character of tonoplast from Hippeastrum vacuoles

    Energy Technology Data Exchange (ETDEWEB)

    Wagner, G.J.

    1981-01-01

    The membrane of anthocyanin containing Hippeastrum petal vacuoles was examined for protein and enzyme content after purification by equilibrium density centrifugation. Light scattering, protein, and a Mg/sup 2 +/ -dependent nucleotide specific ATPase were associated with membrane having a density of 1.08 to 1.12 grams per cubic centimeter. A small amount of acid phosphatase was also present in this region of the gradient, but this activity peaked at about 1.12 grams per cubic centimeter. A component of yeast tonoplast, ..cap alpha..-mannosidase, was not significantly present. UDP-glucose, anthocyanidin-3-O-glucosyltransferase, thought to be a cytosol enzyme in Hippeastrum, was absent from tonoplast of vacuoles isolated by osmotic shock in 0.2 molar K/sub 2/HPO/sub 4/ or 0.35 molar mannitol. Vacuolar acid phosphatase was insensitive to ethylenediaminetetraacetate but was 80% inhibited by 10 millimolar KF, while ATPase was inactivated by 2 millimolar ethylenediaminetetraacetate and only 50% inhibited by 10 millimolar KF,. Five major and about 9 minor polypeptides were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of membrane protein on 5 to 30 and 6 to 16% gradient gels.

  12. Redox Enzymes of Red Beetroot Vacuoles (Beta vulgaris L.

    Directory of Open Access Journals (Sweden)

    E.V. Pradedova

    2014-12-01

    Full Text Available Years of research have shown that some of the redox elements (enzymes, coenzymes, and co-substrate are isolated from each other kinetic and spatial manner (compartmentalization in the eukaryotic cells. The redox elements forming the "highly" and "widely" specialized redox system are found in all cell structures: mitochondria, plastids, peroxisomes, apoplast, nucleus etc. In recent years the active involvement of the central vacuole in the maintenance of the plant cell redox homeostasis is discussed, actually the information about the vacuolar redox system is very small. The high-priority redox processes and "redox-specialization" of the vacuolar compartment are not known. We have begun a study of red beet-root vacuole redox systems (Beta vulgaris L. and have identified redox enzymes such as: phenol peroxidase (EC 1.11.1.7, superoxide dismutase (EC 1.15.1.1 and glutathione reductase (EC 1.8.1.7. This paper presents some of the characteristics of these enzymes and considers the probable ways of their functioning in vacuolar redox chains.

  13. Reactive oxygen species (ROS) is not a promotor of taxol-induced cytoplasmic vacuolization

    Science.gov (United States)

    Sun, Qingrui; Chen, Tongsheng

    2009-02-01

    we have previously reported that taxol, a potent anticancer agent, induces caspase-independent cell death and cytoplasmic vacuolization in human lung adenocarcinoma (ASTC-a-1) cells. However, the mechanisms of taxol-induced cytoplasmic vacuolization are poorly understood. Reactive oxygen species (ROS) has been reported to be involved in the taxol-induced cell death. Here, we employed confocal fluorescence microscopy imaging to explore the role of ROS in taxol-induced cytoplasmic vacuolization. We found that ROS inhibition by addition of N-acetycysteine (NAC), a total ROS scavenger, did not suppress these vacuolization but instead increased vacuolization. Take together, our results showed that ROS is not a promotor of the taxol-induced cytoplasmic vacuolization.

  14. Simplification of vacuole structure during plant cell death triggered by culture filtrates of Erwinia carotovora

    Institute of Scientific and Technical Information of China (English)

    Yumi Hirakawa; Toshihisa Nomura; Seiichiro Hasezawa; Takumi Higaki

    2015-01-01

    Vacuoles are suggested to play crucial roles in plant defense-related cel death. During programmed cel death, previous live cel imaging studies have observed vacuoles to become simpler in structure and have implicated this simplification as a prelude to the vacuole’s rupture and consequent lysis of the plasma membrane. Here, we examined dynamics of the vacuole in cel cycle-synchronized tobacco BY-2 (Nicotiana tabacum L. cv. Bright Yel ow 2) cel s during cel death induced by application of culture filtrates of Erwinia carotovora. The filtrate induced death in about 90%of the cel s by 24 h. Prior to cel death, vacuole shape simplified and endoplasmic actin filaments disassembled;however, the vacuoles did not rupture until after plasma membrane integrity was lost. Instead of facilitating rupture, the simplification of vacuole structure might play a role in the retrieval of membrane components needed for defense-related cel death.

  15. Co-occurrence of tannin and tannin-less vacuoles in sensitive plants.

    Science.gov (United States)

    Fleurat-Lessard, Pierrette; Béré, Emile; Lallemand, Magali; Dédaldéchamp, Fabienne; Roblin, Gabriel

    2016-05-01

    Vacuoles of different types frequently coexist in the same plant cell, but the duality of the tannin/tannin-less vacuoles observed in Mimosa pudica L. is rare. In this plant, which is characterized by highly motile leaves, the development and original features of the double vacuolar compartment were detailed in primary pulvini from the young to the mature leaf stage. In young pulvini, the differentiation of tannin vacuoles first occurred in the epidermis and progressively spread toward the inner cortex. In motor cells of nonmotile pulvini, tannin deposits first lined the membranes of small vacuole profiles and then formed opaque clusters that joined together to form a large tannin vacuole (TV), the proportion of which in the cell was approximately 45%. At this stage, transparent vacuole profiles were rare and small, but as the parenchyma cells enlarged, these profiles coalesced to form a transparent vacuole with a convexity toward the larger-sized tannin vacuole. When leaf motility began to occur, the two vacuole types reached the same relative proportion (approximately 30%). Finally, in mature cells displaying maximum motility, the large transparent colloidal vacuole (CV) showed a relative proportion increasing to approximately 50%. At this stage, the proportion of the tannin vacuole, occurring in the vicinity of the nucleus, decreased to approximately 10%. The presence of the condensed type of tannins (proanthocyanidins) was proven by detecting their fluorescence under UV light and by specific chemical staining. This dual vacuolar profile was also observed in nonmotile parts of M. pudica (e.g., the petiole and the stem). Additional observations of leaflet pulvini showing more or less rapid movements showed that this double vacuolar structure was present in certain plants (Mimosa spegazzinii and Desmodium gyrans), but absent in others (Albizzia julibrissin, Biophytum sensitivum, and Cassia fasciculata). Taken together, these observations strongly suggest that a

  16. Trypanosoma cruzi Differentiates and Multiplies within Chimeric Parasitophorous Vacuoles in Macrophages Coinfected with Leishmania amazonensis.

    Science.gov (United States)

    Pessoa, Carina Carraro; Ferreira, Éden Ramalho; Bayer-Santos, Ethel; Rabinovitch, Michel; Mortara, Renato Arruda; Real, Fernando

    2016-05-01

    The trypanosomatids Leishmania amazonensis and Trypanosoma cruzi are excellent models for the study of the cell biology of intracellular protozoan infections. After their uptake by mammalian cells, the parasitic protozoan flagellates L. amazonensis and T. cruzi lodge within acidified parasitophorous vacuoles (PVs). However, whereas L. amazonensis develops in spacious, phagolysosome-like PVs that may enclose numerous parasites, T. cruzi is transiently hosted within smaller vacuoles from which it soon escapes to the host cell cytosol. To investigate if parasite-specific vacuoles are required for the survival and differentiation of T. cruzi, we constructed chimeric vacuoles by infection of L. amazonensis amastigote-infected macrophages with T. cruzi epimastigotes (EPIs) or metacyclic trypomastigotes (MTs). These chimeric vacuoles, easily observed by microscopy, allowed the entry and fate of T. cruzi in L. amazonensis PVs to be dynamically recorded by multidimensional imaging of coinfected cells. We found that although T. cruzi EPIs remained motile and conserved their morphology in chimeric vacuoles, T. cruzi MTs differentiated into amastigote-like forms capable of multiplying. These results demonstrate that the large adaptive vacuoles of L. amazonensis are permissive to T. cruzi survival and differentiation and that noninfective EPIs are spared from destruction within the chimeric PVs. We conclude that T. cruzi differentiation can take place in Leishmania-containing vacuoles, suggesting this occurs prior to their escape into the host cell cytosol.

  17. Preparative Procedures Markedly Influence the Appearance and Structural Integrity of Protein Storage Vacuoles in Soybean Seeds

    Science.gov (United States)

    In legumes, vacuoles serve as the final depository for storage proteins. The protein storage vacuoles (PSVs) of soybean contain electron-transparent globoid regions in which phytic acid (myo-inositol-1,2,3,4,5,6-hexakisphosphate) is sequestered. Here, I report the effect of preparative procedures o...

  18. Motile tubular vacuoles in extramatrical mycelium and sheath hyphae of ectomycorrhizal systems.

    Science.gov (United States)

    Allaway, W G; Ashford, A E

    2001-01-01

    Extramatrical mycelium and outer hyphae of the sheath of Eucalyptus pilularis-Pisolithus tinctorius mycorrhizas contain abundant motile tubular vacuoles which accumulate the carboxyfluorescein analogue Oregon Green 488 carboxylic acid. The fluorochrome accumulates in a system of small vacuoles, tubules, and larger vacuoles, which are interlinked, motile, and pleiomorphic, in external hyphae, cords, and hyphae of the outer sheath. There is often a difference in fluorescence between two neighbouring cells, indicating that the dolipore septum exercises control on the movement of material between cells. Generally the motile tubular vacuole system in mycorrhizas resembles that previously found in isolated mycelium. The majority of fungal cells in the sheath contain no fluorochrome even after long exposure of the mycorrhiza to the solution, but with differential interference optics the cells are clearly seen to be alive and to contain vacuoles resembling those in the outer hyphae. In translocation experiments, long-distance transport of the fluorochrome is slow and slight, or even nonexistent in some cases.

  19. Early cytoplasmic vacuolization of African green monkey kidney cells by SV40.

    Science.gov (United States)

    Miyamura, T; Kitahara, T

    1975-01-01

    As early as 3--4 hours after infection with SV40 at a high input multiplicity, African green monkey (Cercopithecus aethiops) kidney (AGMK) cells developed cytoplasmic vacuolization. At 10--20 hours after infection, the vacuolization reached its maximal level, then disappeared and SV40 specific cytopathic change followed. This vacuolization developed before the synthesis of the specific T and V antigens. This early cytoplasmic vacuolization (ECV) was prevented by preincubating the virus with specific antiserum, or by heating the virus with MgCl2. The ECV could be induced by UV-irradiated SV40. Addition of metabolic inhibitors had no effect on the induction of the ECV. These results suggest that the capacity to induce the ECV resides in a structural component(s) of SV40 virion and the vacuolization is not associated with the replication of SV40.

  20. Early cytoplasmic vacuolization of African green monkey kidney cells by SV40. [uv radiation

    Energy Technology Data Exchange (ETDEWEB)

    Miyamura, T.; Kitahara, T.

    1975-01-01

    As early as 3 to 4 hours after infection with SV 40 at a high input multiplicity, African green monkey (Cercopithecus aethiops) kidney (AGMK) cells developed cytoplasmic vacuolization. At 10 to 20 hours after infection, the vacuolization reached its maximal level, then disappeared and SV 40 specific cytopathic change followed. This vacuolization developed before the synthesis of the specific T and V antigens. This early cytoplasmic vacuolization (ECV) was prevented by pre-incubating the virus with specific antiserum, or by heating the virus with MgCl/sub 2/. The ECV could be induced by uv-irradiated SV 40. Addition of metabolic inhibitors had no effect on the induction of the ECV. These results suggest that the capacity to induce the ECV resides in a structural component(s) of SV 40 virion and the vacuolization is not associated with the replication of SV 40.

  1. The nature of human sperm head vacuoles: a systematic literature review.

    Science.gov (United States)

    Boitrelle, Florence; Guthauser, Bruno; Alter, Laura; Bailly, Marc; Wainer, Robert; Vialard, François; Albert, Martine; Selva, Jacqueline

    2013-01-01

    Motile sperm organelle morphology examination (MSOME) involves the use of differential interference contrast microscopy (also called Nomarski contrast) at high magnification (at least 6300x) to improve the observation of live human spermatozoa. In fact, this technique evidences sperm head vacuoles that are not necessarily seen at lower magnifications - particularly if the vacuoles are small (i.e. occupying nature. In an attempt to clarify this debate, we performed a systematic literature review in accordance with the PRISMA guidelines. The PubMed database was searched from 2001 onwards with the terms "MSOME", "human sperm vacuoles", "high-magnification, sperm". Out of 180 search results, 21 relevant English-language publications on the nature of human sperm head vacuoles were finally selected and reviewed. Our review of the literature prompted us to conclude that sperm-head vacuoles are nuclear in nature and are related to chromatin condensation failure and (in some cases) sperm DNA damage.

  2. Organelle Size Scaling of the Budding Yeast Vacuole by Relative Growth and Inheritance.

    Science.gov (United States)

    Chan, Yee-Hung M; Reyes, Lorena; Sohail, Saba M; Tran, Nancy K; Marshall, Wallace F

    2016-05-09

    It has long been noted that larger animals have larger organs compared to smaller animals of the same species, a phenomenon termed scaling [1]. Julian Huxley proposed an appealingly simple model of "relative growth"-in which an organ and the whole body grow with their own intrinsic rates [2]-that was invoked to explain scaling in organs from fiddler crab claws to human brains. Because organ size is regulated by complex, unpredictable pathways [3], it remains unclear whether scaling requires feedback mechanisms to regulate organ growth in response to organ or body size. The molecular pathways governing organelle biogenesis are simpler than organogenesis, and therefore organelle size scaling in the cell provides a more tractable case for testing Huxley's model. We ask the question: is it possible for organelle size scaling to arise if organelle growth is independent of organelle or cell size? Using the yeast vacuole as a model, we tested whether mutants defective in vacuole inheritance, vac8Δ and vac17Δ, tune vacuole biogenesis in response to perturbations in vacuole size. In vac8Δ/vac17Δ, vacuole scaling increases with the replicative age of the cell. Furthermore, vac8Δ/vac17Δ cells continued generating vacuole at roughly constant rates even when they had significantly larger vacuoles compared to wild-type. With support from computational modeling, these results suggest there is no feedback between vacuole biogenesis rates and vacuole or cell size. Rather, size scaling is determined by the relative growth rates of the vacuole and the cell, thus representing a cellular version of Huxley's model.

  3. Active vacuolar H+ ATPase and functional cycle of Rab5 are required for the vacuolation defect triggered by PtdIns(3,5)P2 loss under PIKfyve or Vps34 deficiency.

    Science.gov (United States)

    Compton, Lauren M; Ikonomov, Ognian C; Sbrissa, Diego; Garg, Puneet; Shisheva, Assia

    2016-09-01

    The two evolutionarily conserved mammalian lipid kinases Vps34 and PIKfyve are involved in an important physiological relationship, whereby the former produces phosphatidylinositol (PtdIns) 3P that is used as a substrate for PtdIns(3,5)P2 synthesis by the latter. Reduced production of PtdIns(3,5)P2 in proliferating mammalian cells is phenotypically manifested by the formation of multiple translucent cytoplasmic vacuoles, readily rescued upon exogenous delivery of PtdIns(3,5)P2 or overproduction of PIKfyve. Although the aberrant vacuolation phenomenon has been frequently used as a sensitive functional measure of localized PtdIns(3,5)P2 reduction, cellular factors governing the appearance of cytoplasmic vacuoles under PtdIns3P-PtdIns(3,5)P2 loss remain elusive. To gain further mechanistic insight about the vacuolation process following PtdIns(3,5)P2 reduction, in this study we sought for cellular mechanisms required for manifestation of the aberrant endomembrane vacuoles triggered by PIKfyve or Vps34 dysfunction. The latter was achieved by various means such as pharmacological inhibition, gene disruption, or dominant-interference in several proliferating mammalian cell types. We report here that inhibition of V-ATPase with bafilomycin A1 as well as inactivation of the GTP-GDP cycle of Rab5a GTPase phenotypically rescued or completely precluded the cytoplasmic vacuolization despite the continued presence of inactivated PIKfyve or Vps34. Bafilomycin A1 also restored the aberrant EEA1-positive endosomes, enlarged upon short PIKfyve inhibition with YM201636. Together, our work identifies for the first time that factors such as active V-ATPase or functional Rab5a cycle are acting coincidentally with the PtdIns(3,5)P2 reduction in triggering formation of aberrant cytoplasmic vacuoles under PIKfyve or Vps34 dysfunction.

  4. Phytochelatin-metal(loid) transport into vacuoles shows different substrate preferences in barley and Arabidopsis.

    Science.gov (United States)

    Song, Won-Yong; Mendoza-Cózatl, David G; Lee, Youngsook; Schroeder, Julian I; Ahn, Sang-Nag; Lee, Hyun-Sook; Wicker, Thomas; Martinoia, Enrico

    2014-05-01

    Cadmium (Cd) and arsenic (As) are toxic to all living organisms, including plants and humans. In plants, Cd and As are detoxified by phytochelatins (PCs) and metal(loid)-chelating peptides and by sequestering PC-metal(loid) complexes in vacuoles. Consistent differences have been observed between As and Cd detoxification. Whereas chelation of Cd by PCs is largely sufficient to detoxify Cd, As-PC complexes must be sequestered into vacuoles to be fully detoxified. It is not clear whether this difference in detoxification pathways is ubiquitous among plants or varies across species. Here, we have conducted a PC transport study using vacuoles isolated from Arabidopsis and barley. Arabidopsis vacuoles accumulated low levels of PC2 -Cd, and vesicles from yeast cells expressing either AtABCC1 or AtABCC2 exhibited negligible PC2 -Cd transport activity compared with PC2 -As. In contrast, barley vacuoles readily accumulated comparable levels of PC2 -Cd and PC2 -As. PC transport in barley vacuoles was inhibited by vanadate, but not by ammonium, suggesting the involvement of ABC-type transporters. Interestingly, barley vacuoles exhibited enhanced PC2 transport activity when essential metal ions, such as Zn(II), Cu(II) and Mn(II), were added to the transport assay, suggesting that PCs might contribute to the homeostasis of essential metals and detoxification of non-essential toxic metal(loid)s.

  5. The Water to Solute Permeability Ratio Governs the Osmotic Volume Dynamics in Beetroot Vacuoles

    Science.gov (United States)

    Vitali, Victoria; Sutka, Moira; Amodeo, Gabriela; Chara, Osvaldo; Ozu, Marcelo

    2016-01-01

    Plant cell vacuoles occupy up to 90% of the cell volume and, beyond their physiological function, are constantly subjected to water and solute exchange. The osmotic flow and vacuole volume dynamics relies on the vacuole membrane -the tonoplast- and its capacity to regulate its permeability to both water and solutes. The osmotic permeability coefficient (Pf) is the parameter that better characterizes the water transport when submitted to an osmotic gradient. Usually, Pf determinations are made in vitro from the initial rate of volume change, when a fast (almost instantaneous) osmolality change occurs. When aquaporins are present, it is accepted that initial volume changes are only due to water movements. However, in living cells osmotic changes are not necessarily abrupt but gradually imposed. Under these conditions, water flux might not be the only relevant driving force shaping the vacuole volume response. In this study, we quantitatively investigated volume dynamics of isolated Beta vulgaris root vacuoles under progressively applied osmotic gradients at different pH, a condition that modifies the tonoplast Pf. We followed the vacuole volume changes while simultaneously determining the external osmolality time-courses and analyzing these data with mathematical modeling. Our findings indicate that vacuole volume changes, under progressively applied osmotic gradients, would not depend on the membrane elastic properties, nor on the non-osmotic volume of the vacuole, but on water and solute fluxes across the tonoplast. We found that the volume of the vacuole at the steady state is determined by the ratio of water to solute permeabilites (Pf/Ps), which in turn is ruled by pH. The dependence of the permeability ratio on pH can be interpreted in terms of the degree of aquaporin inhibition and the consequently solute transport modulation. This is relevant in many plant organs such as root, leaves, cotyledons, or stems that perform extensive rhythmic growth movements

  6. The water to solute permeability ratio governs the osmotic volume dynamics in beetroot vacuoles

    Directory of Open Access Journals (Sweden)

    Victoria Vitali

    2016-09-01

    Full Text Available Plant cell vacuoles occupy up to 90% of the cell volume and, beyond their physiological function, are constantly subjected to water and solute exchange. The osmotic flow and vacuole volume dynamics relies on the vacuole membrane -the tonoplast- and its capacity to regulate its permeability to both water and solutes. The osmotic permeability coefficient (Pf is the parameter that better characterizes the water transport when submitted to an osmotic gradient. Usually, Pf determinations are made in vitro from the initial rate of volume change, when a fast (almost instantaneous osmolality change occurs. When aquaporins are present, it is accepted that initial volume changes are only due to water movements. However, in living cells osmotic changes are not necessarily abrupt but gradually imposed. Under these conditions, water flux might not be the only relevant driving force shaping the vacuole volume response. In this study, we quantitatively investigated volume dynamics of isolated Beta vulgaris root vacuoles under progressively applied osmotic gradients at different pH, a condition that modifies the tonoplast Pf. We followed the vacuole volume changes while simultaneously determining the external osmolality time-courses and analyzing these data with mathematical modelling. Our findings indicate that vacuole volume changes, under progressively applied osmotic gradients, would not depend on the membrane elastic properties, nor on the non-osmotic volume of the vacuole, but on water and solute fluxes across the tonoplast. We found that the volume of the vacuole at the steady state is determined by the ratio of water to solute permeabilites (Pf/Ps, which in turn is ruled by pH. The dependence of the permeability ratio on pH can be interpreted in terms of the degree of aquaporin inhibition and the consequently solute transport modulation. This is relevant in many plant organs such as root, leaves, cotyledons or stems that perform extensive rhythmic

  7. Content and vacuole extravacuole distribution of neutral sugars, free amino acids, and anthocyanin in protoplasts

    Energy Technology Data Exchange (ETDEWEB)

    Wagner, G.J.

    1979-07-01

    Neutral sugar, free amino acid, and anthocyanin levels and vacuole/extravacuole distribution were determined for Hippeastrum and Tulipa petal and Tulipa leaf protoplasts. Glucose and fructose, the predominant neutral monosaccharides observed, were primarily vacuolar in location. Glutamine, the predominant free amino acid found, was primarily extravacuolar. ..gamma..-methyleneglutamate was identified as a major constituent of Tulipa protoplasts. Qualitative characterization of Hippeastrum petal and vacuole organic acids indicated the presence of oxalic, malic, citric, and isocitric acids. Data are presented which indicate that vacuoles obtained by gentle osmotic shock of protoplasts in dibasic phosphate have good purity and retain their contents.

  8. Glutathione Reductase of Vacuole. Comparison of Glutathione Reductase Activity of Vacuole and Tissue Extract of Red Beet Root (Beta vulgaris L.

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    E.V. Pradedova

    2016-02-01

    Full Text Available Glutathione reductase (GR, EC 1.8.1.7 is the enzyme that reduces oxidized glutathione (GSSG and thus regulates the redox state of glutathione (GSH/GSSG. GR has been studied in most plants. This enzyme has been identified in chloroplasts and cytosol, so these cellular compartments are considered to be the main place of the enzyme localization. In the same time, just a little is known about GR vacuoles. There are no conclusive evidences to prove the presence or absence of this enzyme in the vacuoles. GR activity was found in the vacuoles of red beet root cells (Beta vulgaris L.. The level of activity, the optimum pH and isoenzyme composition of GR were compared in the vacuoles and tissue extract of beet root. Vacuolar GR activity was quite high, it was 1.5-2 times higher than the activity of the tissue extract. Enzyme pH optimum of all the objects were identical. pH-optimum depend on the pyridine nucleotide nature: pH 7.0-8.0 was an optimal range with NADPH; pH 5.0 – with NADH. GR activity of the vacuoles and tissue extracts decreased in the presence of a noncompetitive inhibitor 1-chloro-2.4-dinitrobenzene (CDNB, indicating the specificity of this enzymatic reaction. Two bands with glutathione reductase activity have been identified in the vacuoles and tissue extracts using zymography method to determine the enzymatic activity in PAAG after electrophoresis of proteins. Belonging to the GR isoforms of these bands was confirmed by enzyme immunoassay (Western blotting. The electric mobility of isoforms of the study objects did not differ significantly. It is concluded that the biochemical characteristics of vacuolar glutathione reductase were substantially identical to the biochemical characteristics of other localization GR.

  9. The Proteome of the Isolated Chlamydia trachomatis Containing Vacuole Reveals a Complex Trafficking Platform Enriched for Retromer Components.

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    Lukas Aeberhard

    2015-06-01

    Full Text Available Chlamydia trachomatis is an important human pathogen that replicates inside the infected host cell in a unique vacuole, the inclusion. The formation of this intracellular bacterial niche is essential for productive Chlamydia infections. Despite its importance for Chlamydia biology, a holistic view on the protein composition of the inclusion, including its membrane, is currently missing. Here we describe the host cell-derived proteome of isolated C. trachomatis inclusions by quantitative proteomics. Computational analysis indicated that the inclusion is a complex intracellular trafficking platform that interacts with host cells' antero- and retrograde trafficking pathways. Furthermore, the inclusion is highly enriched for sorting nexins of the SNX-BAR retromer, a complex essential for retrograde trafficking. Functional studies showed that in particular, SNX5 controls the C. trachomatis infection and that retrograde trafficking is essential for infectious progeny formation. In summary, these findings suggest that C. trachomatis hijacks retrograde pathways for effective infection.

  10. Vibrio cholerae hemolysin is required for lethality, developmental delay, and intestinal vacuolation in Caenorhabditis elegans.

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    Hediye Nese Cinar

    Full Text Available BACKGROUND: Cholera toxin (CT and toxin-co-regulated pili (TCP are the major virulence factors of Vibrio cholerae O1 and O139 strains that contribute to the pathogenesis of disease during devastating cholera pandemics. However, CT and TCP negative V. cholerae strains are still able to cause severe diarrheal disease in humans through mechanisms that are not well understood. METHODOLOGY/PRINCIPAL FINDINGS: To determine the role of other virulence factors in V. cholerae pathogenesis, we used a CT and TCP independent infection model in the nematode Caenorhabditis elegans and identified the hemolysin A (hlyA gene as a factor responsible for animal death and developmental delay. We demonstrated a correlation between the severity of infection in the nematode and the level of hemolytic activity in the V. cholerae biotypes. At the cellular level, V. cholerae infection induces formation of vacuoles in the intestinal cells in a hlyA dependent manner, consistent with the previous in vitro observations. CONCLUSIONS/SIGNIFICANCE: Our data strongly suggest that HlyA is a virulence factor in C. elegans infection leading to lethality and developmental delay presumably through intestinal cytopathic changes.

  11. Localization of acid hydrolases in protoplasts. Examination of the proposed lysosomal function of the mature vacuole

    Energy Technology Data Exchange (ETDEWEB)

    Butcher, H.C.; Wagner, G.J.; Siegelman, H.W.

    1977-06-01

    The development of techniques to isolate and purify relatively large quantities of intact vacuoles from mature tissues permits direct biochemical analysis of this ubiquitous mature plant cell organelle. Vacuoles and a fraction enriched in soluble cytoplasmic constituents were quantitatively prepared from Hippeastrum flower petal protoplasts. Vacuolar lysate and soluble cytoplasmic fractions were examined for acid hydrolase activities commonly associated with animal lysosomes, and pH optima were determined. Esterase, protease, carboxypeptidase, ..beta..-galactosidase, ..cap alpha..-glycosidase and ..beta..-glycosidase, not found in the vacuole lysate fraction, were components of the soluble cytoplasmic fraction. Acid phosphatase, RNase and DNase were present in both fractions. Vacuolar enzyme activities were also examined as a function of flower development from bud through senescent stages. The data obtained are not consistent with the concept that the mature plant cell vacuole functions as a generalized lysosome.

  12. A soluble acid invertase is directed to the vacuole by a signal anchor mechanism.

    Science.gov (United States)

    Rae, Anne L; Casu, Rosanne E; Perroux, Jai M; Jackson, Mark A; Grof, Christopher P L

    2011-06-15

    Enzyme activities in the vacuole have an important impact on the net concentration of sucrose. In sugarcane (Saccharum hybrid), immunolabelling demonstrated that a soluble acid invertase (β-fructofuranosidase; EC 3.2.1.26) is present in the vacuole of storage parenchyma cells during sucrose accumulation. Examination of sequences from sugarcane, barley and rice showed that the N-terminus of the invertase sequence contains a signal anchor and a tyrosine motif, characteristic of single-pass membrane proteins destined for lysosomal compartments. The N-terminal peptide from the barley invertase was shown to be capable of directing the green fluorescent protein to the vacuole in sugarcane cells. The results suggest that soluble acid invertase is sorted to the vacuole in a membrane-bound form.

  13. Oculopharyngeal Weakness, Hypophrenia, Deafness, and Impaired Vision: A Novel Autosomal Dominant Myopathy with Rimmed Vacuoles

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    Ting Chen

    2016-01-01

    Conclusions: We reported a novel autosomal dominant myopathy with rimmed vacuoles characterized by dysarthria, dysphagia, external ophthalmoplegia, limb weakness, hypophrenia, deafness, and impaired vision, but the causative gene has not been found and needs further study.

  14. THE TONOPLAST TRANSPORT SYSTEMS OF PLANT VACUOLES AND THEIR POTENTIAL APPLICATION IN BIOTECHNOLOGY

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    S. V. Isayenkov

    2013-06-01

    Full Text Available The pivotal role of plant vacuoles in plant survival was discussed in the review. Particularly, the providing of cellular turgor, accumulation of inorganic osmolytes and nutrients are the primary tasks of these cellular organelles. The main mechanisms of tonoplast transport systems were described. The known transport pathways of minerals, heavy metals, vitamins and other organic compounds were classified and outlined. The main systems of membrane vacuolar transport were reviewed. The outline of the physiological functions and features of vacuolar membrane transport proteins were performed. The physiological role of transport of minerals, nutrients and other compounds into vacuoles were discussed. This article reviews the main types of plant vacuoles and their functional role in plant cell. Current state and progress in vacuolar transport research was outlined. The examples of application for rinciples and mechanisms of vacuolar membrane transport in plant biotechnology were iven. The perspectives and approaches in plant and food biotechnology concerning transport and physiology of vacuoles are discussed.

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

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    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.

  16. Enhanced Membrane Fusion in Sterol-enriched Vacuoles Bypasses the Vrp1p RequirementD⃞

    OpenAIRE

    Tedrick, Kelly; Trischuk, Tim; Lehner, Richard; Eitzen, Gary

    2004-01-01

    Organization of lipids into membrane microdomains is a vital mechanism of protein processing. Here we show that overexpression of ERG6, a gene involved in ergosterol synthesis, elevates sterol levels 1.5-fold on the vacuole membrane and enhances their homotypic fusion. The mechanism of sterol-enhanced fusion is not via more efficient sorting, but instead promotes increased kinetics of fusion subreactions. We initially isolated ERG6 as a suppressor of a vrp1Δ growth defect selective for vacuol...

  17. Membrane-bound ATPase of intact vacuoles and tonoplasts isolated from mature plant tissue

    Energy Technology Data Exchange (ETDEWEB)

    Lin, W.; Wagner, G.J.; Siegelman, H.W.; Hind, G.

    1977-01-01

    Intact vacuoles were isolated from petals of Hippeastrum and Tulipa (Wagner, G. J. and Siegelman, H. W. (1975) Science 190, 1298 to 1299). The ATPase activity of fresh vacuole suspensions was found to be 2 to 3 times that of protoplasts from the same tissue. 70 to 80% of the ATPase activity of intact vacuoles was recovered in tonoplast preparations. The antibiotic Dio-9 at 6 ..mu..g/10/sup 6/ vacuoles or protoplasts causes 40% inhibition. However, only the protoplast ATPase is sensitive to oligomycin. N,N'-dicyclohexylcarbondiimide (DCCD) slightly stimulates ATPase activity in both vacuole and protoplast suspensions, whereas ethyl-3-(3-dimethylaminopropyl carbodiimide) (EDAC) strongly inhibits. Spectrophotometric studies show that in the petal the vacuolar contents have a pH of 4.0 for Tulipa and 4.3 for Hippeastrum, whereas the intact isolated vacuole has an internal pH of 7.0 (in pH 8.0 buffer) for Tulipa and about 7.3 for Hippeastrum. Internal ion concentrations of 150, 46, 30, 30 and 6 mM were found for K/sup +/, Na/sup +/, Mg/sup 2 +/, Cl/sup -/, and Ca/sup 2 +/ respectively, which are about the same as those in protoplasts.

  18. Vacuolating cytotoxin A (VacA) - A multi-talented pore-forming toxin from Helicobacter pylori.

    Science.gov (United States)

    Junaid, Muhammad; Linn, Aung Khine; Javadi, Mohammad Bagher; Al-Gubare, Sarbast; Ali, Niaz; Katzenmeier, Gerd

    2016-08-01

    Helicobacter pylori is associated with severe and chronic diseases of the stomach and duodenum such as peptic ulcer, non-cardial adenocarcinoma and gastric lymphoma, making Helicobacter pylori the only bacterial pathogen which is known to cause cancer. The worldwide rate of incidence for these diseases is extremely high and it is estimated that about half of the world's population is infected with H. pylori. Among the bacterial virulence factors is the vacuolating cytotoxin A (VacA), which represents an important determinant of pathogenicity. Intensive characterization of VacA over the past years has provided insight into an ample variety of mechanisms contributing to host-pathogen interactions. The toxin is considered as an important target for ongoing research for several reasons: i) VacA displays unique features and structural properties and its mechanism of action is unrelated to any other known bacterial toxin; ii) the toxin is involved in disease progress and colonization by H. pylori of the stomach; iii) VacA is a potential and promising candidate for the inclusion as antigen in a vaccine directed against H. pylori and iv) the vacA gene is characterized by a high allelic diversity, and allelic variants contribute differently to the pathogenicity of H. pylori. Despite the accumulation of substantial data related to VacA over the past years, several aspects of VacA-related activity have been characterized only to a limited extent. The biologically most significant effect of VacA activity on host cells is the formation of membrane pores and the induction of vacuole formation. This review discusses recent findings and advances on structure-function relations of the H. pylori VacA toxin, in particular with a view to membrane channel formation, oligomerization, receptor binding and apoptosis.

  19. Formats

    Directory of Open Access Journals (Sweden)

    Gehmann, Ulrich

    2012-03-01

    Full Text Available In the following, a new conceptual framework for investigating nowadays’ “technical” phenomena shall be introduced, that of formats. The thesis is that processes of formatting account for our recent conditions of life, and will do so in the very next future. It are processes whose foundations have been laid in modernity and which will further unfold for the time being. These processes are embedded in the format of the value chain, a circumstance making them resilient to change. In addition, they are resilient in themselves since forming interconnected systems of reciprocal causal circuits.Which leads to an overall situation that our entire “Lebenswelt” became formatted to an extent we don’t fully realize, even influencing our very percep-tion of it.

  20. Burkholderia cepacia complex isolates survive intracellularly without replication within acidic vacuoles of Acanthamoeba polyphaga.

    Science.gov (United States)

    Lamothe, Julie; Thyssen, Sandra; Valvano, Miguel A

    2004-12-01

    We have previously demonstrated that isolates of the Burkholderia cepacia complex can survive intracellularly in murine macrophages and in free-living Acanthamoeba. In this work, we show that the clinical isolates B. vietnamiensis strain CEP040 and B. cenocepacia H111 survived but did not replicate within vacuoles of A. polyphaga. B. cepacia-containing vacuoles accumulated the fluid phase marker Lysosensor Blue and displayed strong blue fluorescence, indicating that they had low pH. In contrast, the majority of intracellular bacteria within amoebae treated with the V-ATPse inhibitor bafilomycin A1 localized in vacuoles that did not fluoresce with Lysosensor Blue. Experiments using bacteria fluorescently labelled with chloromethylfluorescein diacetate demonstrated that intracellular bacteria remained viable for at least 24 h. In contrast, Escherichia coli did not survive within amoebae after 2 h post infection. Furthermore, intracellular B. vietnamiensis CEP040 retained green fluorescent protein within the bacterial cytoplasm, while this protein rapidly escaped from the cytosol of phagocytized heat-killed bacteria into the vacuolar lumen. Transmission electron microscopy analysis confirmed that intracellular Burkholderia cells were structurally intact. In addition, both Legionella pneumophila- and B. vietnamiensis-containing vacuoles did not accumulate cationized ferritin, a compound that localizes within the lysosome. Thus, our observations support the notion that B. cepacia complex isolates can use amoebae as a reservoir in the environment by surviving without intracellular replication within an acidic vacuole that is distinct from the lysosomal compartment.

  1. "Autophagy suite": Atg9 cycling in the cytoplasm to vacuole targeting pathway.

    Science.gov (United States)

    Munakata, Nobuo; Klionsky, Daniel J

    2010-08-01

    Macroautophagy continues to gather increasing attention because it is connected with a wide range of human pathophysiologies, developmental processes and life span extension. It is also an interesting process from a basic cellular biology standpoint, as it involves dynamic membrane rearrangements and multiple protein-protein interactions. Although macroautophagy can be nonspecific, there are many examples of selective sequestration including pexophagy, mitophagy and the cytoplasm to vacuole targeting (Cvt) pathway. At present, the Cvt pathway is unique in that it is the only example of a biosynthetic use of macroautophagy. Most of the autophagy-related (Atg) proteins are involved in the Cvt pathway, and various types of analyses have placed these proteins at particular stages of the process. For example, Atg9 is the only characterized transmembrane protein that is absolutely required for Cvt vesicle formation, and it is proposed to carry membrane from peripheral donor sites to the phagophore assembly site where the vesicle forms. Additional proteins, including Atg11, Atg23 and Atg27 are involved in this anterograde movement, whereas Atg1-Atg13 and Atg2-Atg18 are required for the retrograde return to the peripheral sites. Even when we illustrate our understanding of these events in a schematic model, however, they are by necessity flat two-dimensional representations, lacking movement and sound. Yet the cell is a living entity that is not well served by this sole method of information display. Accordingly, we decided to present the Cvt pathway as a vibrant, dynamic process by combining science, music and illustration.

  2. Proteome analysis of potato juice and tuber vacuoles from cv. Kuras

    DEFF Research Database (Denmark)

    Jørgensen, Malene

    from the vacuoles of mature potato tuber. A comprehensive investigation of the vacuolar proteome and a detailed analysis of the identified proteins and their possible roles in vacuolar function were carried out. A primary requirement of any proteomic analysis of an organelle is purity of the isolated...... organelle. A method was developed for isolation of highly purified intact vacuoles. The proteome analysis of the purified vacuoles involved separation of native soluble proteins by gel filtration with Superdex 200 into nine fractions. Each fraction was analyzed in two ways, by SDS-PAGE followed by protein...... open reading frames and expressed sequence tag databases were identified.   The putative vacuolar proteins were categorized into nine major functional groups (number of proteins in brackets): storage proteins (8), stress or defense related proteins (102), protein signaling and degradation processes (25...

  3. Disruption of Toxoplasma gondii Parasitophorous Vacuoles by the Mouse p47-Resistance GTPases.

    Directory of Open Access Journals (Sweden)

    2005-11-01

    Full Text Available The p47 GTPases are essential for interferon-gamma-induced cell-autonomous immunity against the protozoan parasite, Toxoplasma gondii, in mice, but the mechanism of resistance is poorly understood. We show that the p47 GTPases, including IIGP1, accumulate at vacuoles containing T. gondii. The accumulation is GTP-dependent and requires live parasites. Vacuolar IIGP1 accumulations undergo a maturation-like process accompanied by vesiculation of the parasitophorous vacuole membrane. This culminates in disruption of the parasitophorous vacuole and finally of the parasite itself. Over-expression of IIGP1 leads to accelerated vacuolar disruption whereas a dominant negative form of IIGP1 interferes with interferon-gamma-mediated killing of intracellular parasites. Targeted deletion of the IIGP1 gene results in partial loss of the IFN-gamma-mediated T. gondii growth restriction in mouse astrocytes.

  4. The Expression of VacA in BCF of Helicobacter Pylori and Its Relationship to Vacuolated Effect

    Institute of Scientific and Technical Information of China (English)

    施理; 侯晓华; 易粹琼; 张锦坤

    2002-01-01

    Summary: The vacuolated effect of Helicobacter (H. Pylori) and its relationship to vacuolated cyto toxin antigen (VacA) were investigated by the method of cytotoxic test and SDS-pobyacrylamide gel electrophoresis (SDS-PAGE). Of the 62 clinical isolates, the broth culture filter (BCF) of 43 strains causecl the Vero cell intracytoplasmically vacuolated. H. Pylori strains were divided into H. Pylori (Toxin+) group with vacuolated effect and H. Pylori (Toxin-) group without vacuolated effect. The analysis of the BCF of H. Pylori (Toxin+) and that of H. Pylori (Toxin-) was studied by SDS-PAGE and Scan reader. A kind of protein with 87 ku molecular weight was recognized in the BCF of 30.23 % (13/43) H. Pylori (Toxin+) strains but in none of that of H. Pylori (Toxin-) strains, the difference was statistically significant (P<0. 05). There was a significant and concordant relation ship between OD of the protein band with 87 ku molecular weight and titer of vacuolated activity of H. Pylori(Toxin+) (r=0. 67 and P<0. 05 by linear regression analysis). H. Pylori strains were di-vided into H. Pylori (Toxin+) group with vacuolated effect and H. Pylori (Toxin-) group without vacuolated effect. The vacuolated effect of H. Pylori (Toxin+) was caused by the protein with 87 ku molecular weight (VacA).

  5. Intratumoral injection of taxol in vivo suppresses A549 tumor showing cytoplasmic vacuolization.

    Science.gov (United States)

    Wang, Chaoyang; Chen, Tongsheng

    2012-04-01

    Based on our recent in vitro studies, this report was designed to explore the mechanism by which high concentration of taxol (70 µM) induced paraptosis-like cell death in human lung carcinoma (A549) cells, and to evaluate the therapeutic efficacy of taxol using A549 tumor-bearing mice in vivo. Exposure of cells to taxol induced time-dependent cytotoxicity and cytoplasmic vacuolization without the involvement of Bax, Bak, Mcl-1, Bcl-XL, and caspase-3. Although taxol treatment induced activating transcription factor 6 (ATF6) cleavage indicative of endoplasmic reticulum (ER) stress, silencing ATF6 by shATF6 did not prevent taxol-induced both cytotoxcity and cytoplasmic vacuolization, suggesting that taxol-induced cytoplasmic vacuolization and cell death were not due to ER stress. Moreover, taxol-treated cells did not show DNA fragmentation and loss of mitochondrial membrane potential, the typical characteristics of apoptosis. In addition, taxol-induced cytoplasmic vacuolization did not show the cellular lysis, the characteristics of oncosis, and positive of β-galactosidase, the characteristic of senescence, indicating that taxol induced paraptosis-like cell death is neither oncosis nor senescence. Moreover, our in vivo data showed that intratumoral injection of taxol (50 mg/kg) in A549 tumor xenograft mice on day 1 and day 19 potently suppressed tumor growth showing significant ER vacuolization without toxicity. In conclusion, high concentration of taxol exhibits a significant anticancer activity by inducing paraptosis-like cell death in vitro and in vivo, without significant toxicity, suggesting a promising therapeutic strategy for apoptosis-resistance cancer by inducing ER vacuolization.

  6. Vacuole formation in mast cells responding to osmotic stress and to F-actin disassembly

    DEFF Research Database (Denmark)

    Koffer, Anna; Williams, Mark; Johansen, Torben

    2002-01-01

    Fluorescent probes were used to visualize the morphology of membranes and of F-actin in rat peritoneal mast cells, exposed to hyperosmotic medium and consequently reversed to isotonicity. Hypertonicity induced cell shrinkage followed by a regulatory volume increase, and cell alkalinization...

  7. Cell-to-cell spread and massive vacuole formation after Cryptococcus neoformans infection of murine macrophages

    OpenAIRE

    Casadevall Arturo; Alvarez Mauricio

    2007-01-01

    Abstract Background The interaction between macrophages and Cryptococcus neoformans (Cn) is critical for containing dissemination of this pathogenic yeast. However, Cn can either lyse macrophages or escape from within them through a process known as phagosomal extrusion. Both events result in live extracellular yeasts capable of reproducing and disseminating in the extracellular milieu. Another method of exiting the intracellular confines of cells is through host cell-to-cell transfer of the ...

  8. Juliprosopine and juliprosine from prosopis juliflora leaves induce mitochondrial damage and cytoplasmic vacuolation on cocultured glial cells and neurons.

    Science.gov (United States)

    Silva, Victor Diogenes A; Pitanga, Bruno P S; Nascimento, Ravena P; Souza, Cleide S; Coelho, Paulo Lucas C; Menezes-Filho, Noélio; Silva, André Mário M; Costa, Maria de Fátima D; El-Bachá, Ramon S; Velozo, Eudes S; Costa, Silvia L

    2013-12-16

    Prosopis juliflora is a shrub largely used for animal and human consumption. However, ingestion has been shown to induce intoxication in animals, which is characterized by neuromuscular alterations induced by mechanisms that are not yet well understood. In this study, we investigated the cytotoxicity of a total alkaloid extract (TAE) and one alkaloid fraction (F32) obtained from P. juliflora leaves to rat cortical neurons and glial cells. Nuclear magnetic resonance characterization of F32 showed that this fraction is composed of a mixture of two piperidine alkaloids, juliprosopine (majority constituent) and juliprosine. TAE and F32 at concentrations between 0.3 and 45 μg/mL were tested for 24 h on neuron/glial cell primary cocultures. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test revealed that TAE and F32 were cytotoxic to cocultures, and their IC50 values were 31.07 and 7.362 μg/mL, respectively. Exposure to a subtoxic concentration of TAE or F32 (0.3-3 μg/mL) induced vacuolation and disruption of the astrocyte monolayer and neurite network, ultrastructural changes, characterized by formation of double-membrane vacuoles, and mitochondrial damage, associated with changes in β-tubulin III and glial fibrillary acidic protein expression. Microglial proliferation was also observed in cultures exposed to TAE or F32, with increasing levels of OX-42-positive cells. Considering that F32 was more cytotoxic than TAE and that F32 reproduced in vitro the main morphologic and ultrastructural changes of "cara torta" disease, we can also suggest that piperidine alkaloids juliprosopine and juliprosine are primarily responsible for the neurotoxic damage observed in animals after they have consumed the plant.

  9. Vibrio cholerae O1 Strains of Different Ribotypes have Similar hlyA RFLP Patterns but Different Vacuolating Ability

    Directory of Open Access Journals (Sweden)

    Jorge E. Vidal

    2007-01-01

    Full Text Available Extensive cytoplasmic vacuolation on Vero and HeLa cells in vitro by the Vibrio cholerae pore forming toxin HlyA, has been previously reported by our group. Vibrio cholerae O1 and non-O1 pathogenic strains show differences in the potential to induce vacuolation, here we study occurring variations on vacuolating cytotoxic ability, related to changes in the nucleotide sequence of the hlyA-orf. A collection of eight toxigenic strains of V. cholerae O1 El Tor and a non-toxigenic one, all belonging to different ribotypes was tested for their vacuolating ability, and hlyA-orf similarity based on PCR and RFLPs. The strains had extremely different vacuolating capacities, those from the ribotype 2 isolated from the US Gulf Coast, showed the highest vacuolating titer (10240 dil, and the rest of the collection had considerably lower titers ranging among 40 to 360 dilutions. PCR of hlyA-orf, was performed and RFLPs were generated using seven restriction enzymes, this approach later revealed small changes of restriction maps, among the strains. The phenogram constructed from the RFLPs, showed two major branches, one of them included most of the strains, the other separates the only Mexican wild type non-O1 Vibrio cholerae. To test for vacuolating ability out of the Vibrio genetic context, the amplified hlyA-orfs from the collection of strains were cloned in pGEMT- vector system and supernatants from the recombinant E coli DH5-, showed no differences on vacuolating titers, the clones always were low producers. Results from the cloning, together with those from the phenogram indicated that the hlyA gene is mainly conserved and the differences on vacuolating activity are unrelated to minute changes seen in the hlyA-orf. Production of high vacuolating titers on Vibrio strains could be due to transcriptional regulation. Whether the high vacuolating titer would be related to increased virulence, is still to be found.

  10. Effects of lysosomal membrane protein depletion on the Salmonella-containing vacuole.

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    Everett A Roark

    Full Text Available Salmonella is an intracellular bacterial pathogen that replicates within a membrane-bound vacuole in host cells. The major lysosomal membrane proteins 1 and 2 (LAMP-1 and LAMP-2 are recruited to the Salmonella-containing vacuole as well as Salmonella- associated filaments (Sifs that emerge from the vacuole. LAMP-1 is a dominant membrane marker for the vacuole and Sifs. Its colocalization with both is dependent on a major secreted bacterial virulence protein, SifA. Here, we show that SifA is required for the recruitment of LAMP-2 and can be used as a second independent marker for both the bacterial vacuolar membrane and Sifs. Further, RNAi studies revealed that in LAMP-1 depleted cells, the bacteria remain membrane bound as measured by their association with LAMP-2 protein. In contrast, LAMP-2 depletion increased the amount of LAMP-1 free bacteria. Together, the data suggests that despite its abundance, LAMP-1 is not essential, but LAMP-2 may be partially important for the Salmonella-containing vacuolar membrane.

  11. Vacuoles in sperm head are not associated with head morphology, DNA damage and reproductive success.

    Science.gov (United States)

    Fortunato, Adriana; Boni, Raffaele; Leo, Rita; Nacchia, Giuseppina; Liguori, Francesca; Casale, Sofia; Bonassisa, Paolo; Tosti, Elisabetta

    2016-02-01

    In this retrospective study of 873 men enrolled for assisted reproduction techniques, relationships between sperm quality parameters, motile sperm organelle morphology examination (MSOME), DNA damage and live birth rate were evaluated. The presence of vacuoles in the sperm heads was detected by MSOME. Either chromatin decondensation or DNA fragmentation was used to study DNA damage. Results show that age significantly affected some of the examined parameters. In particular, sperm concentration was positively correlated (R = 0.088; P = 0.01) and chromatin decondensation was negatively correlated (R = -0.102; P = 0.003) with age. Furthermore, live birth rate was significantly lower in men aged 40 years or older (P fragmentation and live birth rate. Considering sperm heads in relation to the shape (normal/abnormal) and vacuoles (presence/absence), no significant variations in the occurrence of vacuoles in either normal or abnormal heads were found. These data suggest that vacuoles are physiological features that do not alter sperm functionality, and it seems that MSOME is not necessary for increasing the success of assisted reproduction techniques.

  12. Caspase-11 activation requires lysis of pathogen-containing vacuoles by IFN-induced GTPases.

    Science.gov (United States)

    Meunier, Etienne; Dick, Mathias S; Dreier, Roland F; Schürmann, Nura; Kenzelmann Broz, Daniela; Warming, Søren; Roose-Girma, Merone; Bumann, Dirk; Kayagaki, Nobuhiko; Takeda, Kiyoshi; Yamamoto, Masahiro; Broz, Petr

    2014-05-15

    Lipopolysaccharide from Gram-negative bacteria is sensed in the host cell cytoplasm by a non-canonical inflammasome pathway that ultimately results in caspase-11 activation and cell death. In mouse macrophages, activation of this pathway requires the production of type-I interferons, indicating that interferon-induced genes have a critical role in initiating this pathway. Here we report that a cluster of small interferon-inducible GTPases, the so-called guanylate-binding proteins, is required for the full activity of the non-canonical caspase-11 inflammasome during infections with vacuolar Gram-negative bacteria. We show that guanylate-binding proteins are recruited to intracellular bacterial pathogens and are necessary to induce the lysis of the pathogen-containing vacuole. Lysis of the vacuole releases bacteria into the cytosol, thus allowing the detection of their lipopolysaccharide by a yet unknown lipopolysaccharide sensor. Moreover, recognition of the lysed vacuole by the danger sensor galectin-8 initiates the uptake of bacteria into autophagosomes, which results in a reduction of caspase-11 activation. These results indicate that host-mediated lysis of pathogen-containing vacuoles is an essential immune function and is necessary for efficient recognition of pathogens by inflammasome complexes in the cytosol.

  13. Essential domain of receptor tyrosine phosphatase beta (RPTPbeta) for interaction with Helicobacter pylori vacuolating cytotoxin

    DEFF Research Database (Denmark)

    Yahiro, Kinnosuke; Wada, Akihiro; Yamasaki, Eiki

    2004-01-01

    Helicobacter pylori produces a potent exotoxin, VacA, which causes progressive vacuolation as well as gastric injury. Although VacA was able to interact with two receptor-like protein tyrosine phosphatases, RPTPbeta and RPTPalpha, RPTPbeta was found to be responsible for gastric damage caused...

  14. Rimmed vacuoles and the added value of SMI-31 staining in diagnosing sporadic inclusion body myositis.

    Science.gov (United States)

    van der Meulen, M F; Hoogendijk, J E; Moons, K G; Veldman, H; Badrising, U A; Wokke, J H

    2001-07-01

    Problems in diagnosing sporadic inclusion body myositis may arise if all clinical features fit a diagnosis of polymyositis, but the muscle biopsy shows some rimmed vacuoles. Recently, immunohistochemistry with an antibody directed against phosphorylated neurofilament (SMI-31) has been advocated as a diagnostic test for sporadic inclusion body myositis. The aims of the present study were to define a quantitative criterion to differentiate sporadic inclusion body myositis from polymyositis based on the detection of rimmed vacuoles in the haematoxylin-eosin staining and to evaluate the additional diagnostic value of the SMI-31 staining. Based on clinical criteria and creatine kinase levels in patients with endomysial infiltrates, 18 patients complied with the diagnosis of sporadic inclusion body myositis, and 17 with the diagnosis of polymyositis. A blinded observer counted the abnormal fibres in haematoxylin-eosin-stained sections and in SMI-31-stained sections. The optimal cut-off in the haematoxylin-eosin test was 0.3% vacuolated fibres. Adding the SMI-31 staining significantly increased the positive predictive value from 87 to 100%, but increased the negative predictive value only to small extent. We conclude that (1) patients with clinical and laboratory features of polymyositis, including response to treatment, may show rimmed vacuoles in their muscle biopsy and that (2) adding the SMI-31 stain can be helpful in differentiating patients who respond to treatment from patients who do not.

  15. An NPF transporter exports a central monoterpene indole alkaloid intermediate from the vacuole

    DEFF Research Database (Denmark)

    Payne, Richard; Xu, Deyang; Foureau, Emilien

    2017-01-01

    /peptide family (NPF) transporter from Catharanthus roseus, CrNPF2.9, that exports strictosidine, the central intermediate of this pathway, into the cytosol from the vacuole. This discovery highlights the role that intracellular localization plays in specialized metabolism, and sets the stage for understanding...

  16. Retromer and the dynamin Vps1 cooperate in the retrieval of transmembrane proteins from vacuoles

    NARCIS (Netherlands)

    Arlt, Henning; Reggiori, Fulvio; Ungermann, Christian

    2015-01-01

    Endosomes are dynamic organelles that need to combine the ability to successfully deliver proteins and lipids to the lysosome-like vacuole, and recycle others to the Golgi or the plasma membrane. We now show that retromer, which is implicated in retrieval of proteins from endosomes to the Golgi or t

  17. PAS-positive lymphocyte vacuoles can be used as diagnostic screening test for Pompe disease.

    Science.gov (United States)

    Hagemans, Marloes L C; Stigter, Rolinda L; van Capelle, Carine I; van der Beek, Nadine A M E; Winkel, Leon P F; van Vliet, Laura; Hop, Wim C J; Reuser, Arnold J J; Beishuizen, Auke; van der Ploeg, Ans T

    2010-04-01

    Screening of blood films for the presence of periodic acid-Schiff (PAS)-positive lymphocyte vacuoles is sometimes used to support the diagnosis of Pompe disease, but the actual diagnostic value is still unknown. We collected peripheral blood films from 65 untreated Pompe patients and 51 controls. Lymphocyte vacuolization was quantified using three methods: percentage vacuolated lymphocytes, percentage PAS-positive lymphocytes, and a PAS score depending on staining intensity. Diagnostic accuracy of the tests was assessed using receiver operating characteristic (ROC) curves. All three methods fully discerned classic infantile patients from controls. The mean values of patients with milder forms of Pompe disease were significantly higher than those of controls, but full separation was not obtained. The area under the ROC curve was 0.98 for the percentage vacuolated lymphocytes (optimal cutoff value 3; sensitivity 91%, specificity 96%) and 0.99 for the percentage PAS-positive lymphocytes and PAS score (optimal cutoff value 9; sensitivity 100%, specificity 98%). Our data indicate that PAS-stained blood films can be used as a reliable screening tool to support a diagnosis of Pompe disease. The percentage of PAS-positive lymphocytes is convenient for use in clinical practice but should always be interpreted in combination with other clinical and laboratory parameters.

  18. Saccharomyces cerevisiae depend on vesicular traffic between Golgi and vacuole when Inositolphosphorylceramide synthase Aur1 is inactivated

    DEFF Research Database (Denmark)

    Voynova, Natalia S; Roubaty, Carole; Vazquez, Hector M

    2015-01-01

    that vesicle mediated transport between Golgi, endosomes and vacuole becomes crucial for survival when Aur1 is repressed, irrespective of the mode of repression. In addition, vacuolar acidification becomes essential when cells are acutely stressed by AbA, and Quinacrine uptake into vacuoles shows that Ab...

  19. Natural Diversity in the N Terminus of the Mature Vacuolating Cytotoxin of Helicobacter pylori Determines Cytotoxin Activity

    OpenAIRE

    Letley, D. P.; Atherton, J C

    2000-01-01

    Naturally occurring noncytotoxic vacA type s2 strains of Helicobacter pylori have a 12-residue extension to the vacuolating cytotoxin (VacA) compared with cytotoxic type s1 strains. We show that adding the region encoding this extension to type s1 vacA completely abolishes vacuolating cytotoxin activity but has no effect on VacA production.

  20. Retargeting a maize β-glucosidase to the vacuole--evidence from intact plants that zeatin-O-glucoside is stored in the vacuole.

    Science.gov (United States)

    Kiran, Nagavalli S; Benková, Eva; Reková, Alena; Dubová, Jaroslava; Malbeck, Jiří; Palme, Klaus; Brzobohatý, Břetislav

    2012-07-01

    Cytokinin (CK) activity is regulated by the complex interplay of their metabolism, transport, stability and cellular/tissue localization. O-glucosides of zeatin-type CKs are postulated to be storage and/or transport forms. Active CK levels are determined in part by their differential distribution of CK metabolites across different subcellular compartments. We have previously shown that overexpressing chloroplast-localized Zm-p60.1, a maize β-glucosidase capable of releasing active cytokinins from their O- and N3-glucosides, perturbs CK homeostasis in transgenic tobacco. We obtained tobacco (Nicotiana tabacum L., cv Petit Havana SR1) plants overexpressing a recombinant Zm-p60.1 that is targeted to the vacuole. The protein is correctly processed and localized to the vacuole. When grown on medium containing exogenous zeatin, transgenic seedlings rapidly accumulate fresh weight due to ectopic growths at the base of the hypocotyl. The presence of the enzyme in these ectopic structures is shown by histochemical staining. CK quantification reveals that these transgenic seedlings are unable to accumulate zeatin-O-glucoside to levels similar to those observed in the wild type. When crossed with tobacco overexpressing the zeatin-O-glucosyltransferase gene from Phaseolus, the vacuolar variant shows an almost complete reversion in the root elongation assay. This is the first evidence from intact plants that the vacuole is the storage organelle for CK O-glucosides and that they are available to attack by Zm-p60.1. We propose the use of Zm-p60.1 as a robust molecular tool that exploits the reversibility of O-glucosylation and enables delicate manipulations of active CK content at the cellular level.

  1. The Effect of Herbicides on Hydrogen Peroxide Generation in Isolated Vacuoles of Red Beet Root (Beta vulgaris L.

    Directory of Open Access Journals (Sweden)

    E.V. Pradedova

    2015-12-01

    Full Text Available Influence of herbicides on the hydrogen peroxide generation in vacuolar extracts of red beet root (Beta vulgaris L. was investigated. Belonging to different chemical classes of herbicide compounds have been used. Herbicides differ from each other in the mechanism of effects on plants. Clopyralid (aromatic acid herbicide, derivative of picolinic acid and 2.4-D (phenoxyacetic herbicide, characterized by hormone-like effects, contributed to the formation of H2O2 in vacuolar extracts. Fluorodifen (nitrophenyl ether herbicide and diuron (urea herbicide also have increased contents H2O2. These compounds inhibit the electron transport, photosynthesis, and photorespiration in sensitive plants. Herbicidal effect of glyphosate (organophosphorus herbicide is due to the inhibition of amino acid synthesis in plant cells. Glyphosate did not affect the content of H2O2 in vacuolar extracts. Herbicide dependent H2O2-generation did not occur with oxidoreductase inhibitors, potassium cyanide and sodium azide. The results suggest that the formation of ROS in the vacuoles due to activity of oxidoreductases, which could interact with herbicides.

  2. Vacuole/extravacuole distribution of soluble protease in Hippeastrum petal and Triticum leaf protoplasts

    Energy Technology Data Exchange (ETDEWEB)

    Wagner, G.J.; Mulready, P.; Cutt, J.

    1981-11-01

    The subcellular distribution of soluble protease in anthesis-stage, anthocyanin-containing Hippeastrum cv. Dutch Red Hybrid petal protoplasts has been reevaluated and that of Triticum aestivum L. var. Red Coat leaf protoplasts determined using /sup 125/I-fibrin as a protease substrate and improved methods for protoplast and vacuole volume estimation. Results indicate that about 20% of the Hippeastrum petal-soluble protease and about 90% of the wheat leaf-soluble protease can be assigned to the vacuole. Protoplast isolation enzyme labeled with /sup 125/I has been used to assess the efficiency of removing isolation enzyme from protoplasts by repeated washing and by separation of protoplasts from debris using density centrifugation. Results of these studies suggest that protoplasts prepared by both methods retain low levels of isolation enzyme. However, when protoplasts prepared by either method were lysed with washing medium lacking osmoticum, little isolation enzyme contaminated the lysates.

  3. Organization of the cytoplasmic reticulum in the central vacuole of parenchyma cells in Allium cepa L.

    Directory of Open Access Journals (Sweden)

    Tomasz J. Wodzicki

    2015-01-01

    Full Text Available An elaborate and complex cytoplasmic reticulum composed of fine filaments and lamellae ranging from 0.1 to 4 microns in size is revealed by viewing the central vacuole of onion bulb parenchyma cells with the scanning election microscope. The larger cytoplasmic strands, visible with the light microscope, are composed of numerous smaller filaments (some tubular which might explain the observed bidirectional movement of particles in these larger strands. The finely divided cytoplasmic network of filaments is continuous with the parietal cytoplasm inclosing the vacuolar sap. In these highly vacuolated cells the mass of the protoplast is in the form of an intravacuolar reticulum immersed in the cell sap. The probable significance of the vacuolar sap in relation to physiological processes of the cell is discussed.

  4. Mitochondrial Extrusion through the Cytoplasmic Vacuoles during Cell Death*S⃞

    OpenAIRE

    Nakajima, Akihito; Kurihara, Hidetake; Yagita, Hideo; Okumura, Ko; Nakano, Hiroyasu

    2008-01-01

    Under various conditions, noxious stimuli damage mitochondria, resulting in mitochondrial fragmentation; however, the mechanisms by which fragmented mitochondria are eliminated from the cells remain largely unknown. Here we show that cytoplasmic vacuoles originating from the plasma membrane engulfed fragmented mitochondria and subsequently extruded them into the extracellular spaces in undergoing acute tumor necrosis factor α-induced cell death in a caspase-dependent f...

  5. An Acanthamoeba castellanii metacaspase associates with the contractile vacuole and functions in osmoregulation.

    Science.gov (United States)

    Saheb, Entsar; Trzyna, Wendy; Bush, John

    2013-03-01

    Acanthamoeba castellanii is a free-living protozoan. Some strains are opportunistic pathogens. A type-I metacaspase was identified in A. castellanii (Acmcp) and was shown to be expressed through the encystation process. The model organism, Dictyostelium discoideum, has been used here as a model for studying these caspase-like proteins. Separate cell lines expressing a GFP-tagged version of the full length Acmcp protein, as well as a deletion proline region mutant of Acmcp protein (GFP-Acmcp-dpr), have been introduced into D. discoideum. Both mutants affect the cellular metabolism, characterized by an increase in the growth rate. Microscopic imaging revealed an association between Acmcp and the contractile vacuole system in D. discoideum. The treatment of cells with selected inhibitors in different environments added additional support to these findings. This evidence shows that Acmcp plays an important role in contractile vacuole regulation and mediated membrane trafficking in D. discoideum. Additionally, the severe defect in contractile vacuole function in GFP-Acmcp-dpr mutant cells suggests that the proline-rich region in Acmcp has an essential role in binding this protein with other partners to maintain this process. Furthermore, Yeast two-hybrid system identified there are weak interactions of the Dictyostelium contractile vacuolar proteins, including Calmodulin, RabD, Rab11 and vacuolar proton ATPase, with Acmcp protein. Taken together, our findings suggest that A. castellanii metacaspase associate with the contractile vacuole and have an essential role in cell osmoregulation, which contributes to its attractiveness as a possible target for treatment therapies against A. castellanii infection.

  6. Neospora caninum Recruits Host Cell Structures to Its Parasitophorous Vacuole and Salvages Lipids from Organelles.

    Science.gov (United States)

    Nolan, Sabrina J; Romano, Julia D; Luechtefeld, Thomas; Coppens, Isabelle

    2015-05-01

    Toxoplasma gondii and Neospora caninum, which cause the diseases toxoplasmosis and neosporosis, respectively, are two closely related apicomplexan parasites. They have similar heteroxenous life cycles and conserved genomes and share many metabolic features. Despite these similarities, T. gondii and N. caninum differ in their transmission strategies and zoonotic potential. Comparative analyses of the two parasites are important to identify the unique biological features that underlie the basis of host preference and pathogenicity. T. gondii and N. caninum are obligate intravacuolar parasites; in contrast to T. gondii, events that occur during N. caninum infection remain largely uncharacterized. We examined the capability of N. caninum (Liverpool isolate) to interact with host organelles and scavenge nutrients in comparison to that of T. gondii (RH strain). N. caninum reorganizes the host microtubular cytoskeleton and attracts endoplasmic reticulum (ER), mitochondria, lysosomes, multivesicular bodies, and Golgi vesicles to its vacuole though with some notable differences from T. gondii. For example, the host ER gathers around the N. caninum parasitophorous vacuole (PV) but does not physically associate with the vacuolar membrane; the host Golgi apparatus surrounds the N. caninum PV but does not fragment into ministacks. N. caninum relies on plasma lipoproteins and scavenges cholesterol from NPC1-containing endocytic organelles. This parasite salvages sphingolipids from host Golgi Rab14 vesicles that it sequesters into its vacuole. Our data highlight a remarkable degree of conservation in the intracellular infection program of N. caninum and T. gondii. The minor differences between the two parasites related to the recruitment and rearrangement of host organelles around their vacuoles likely reflect divergent evolutionary paths.

  7. Characterization of the Vacuolating Cytotoxin in Helicobacter pylori Strains Isolated from Iran

    Directory of Open Access Journals (Sweden)

    Akbar Oghalaie

    2010-01-01

    Full Text Available Objective: Helicobacter pylori (H. pylori cytotoxin and its heterogeneity amongst strains hasbeen closely linked to the varying infection-associated clinical outcomes. In order to determinethe decisive role of the vacuolating cytotoxin (vacA gene mosaicism in its corresponding geneexpression and phenotype, we aimed to characterize vacA alleles of different H. pylori strainsin addition to the resulting protein and its vacuolating activity in epithelial cell culture.Materials and Methods: vacA gene polymorphism was determined for 80 H. pylori strainsisolated from dyspeptic patients, using multiplex gene-specific polymerase chain reaction(PCR. VacA protein was detected by immuno-blotting assay using a polyclonal anti-VacAantibody. In vitro cytotoxicity assay was conducted on HeLa cells in order to evaluate thevacuolating cytotoxin activity.Results: Genotyping revealed the following strain distribution: 26 (32.5% s1m1, 35(43.8% s1m2, and 19 (23.8% s2m2 subtypes. Infection with s1m1 type strain was significantlyassociated with gastric cancer as compared to non-ulcer dyspepsia (p=0.005and peptic ulcer disease (p=0.008. A 95-kDa immuno-reactive band that represented thevacuolating toxin was demonstrated in SDS-PAGE analysis of concentrated culture filtrate(CCF of H. pylori strains. H. pylori CCFs induced HeLa cell vacuolation which correlatedwith the strain genotype; s1m1 strains demonstrated higher levels of vacuolation as comparedto s1m2 strains, whereas s2m2 strains showed no detectable cytotoxic activity.Conclusion: The current study confirmed the relatively high cytotoxic activity of s1m1type H. pylori strains which infect the majority of patients suffering from gastric cancer andmay be partly responsible for the pathogenesis of this mortal disease.

  8. Oculopharyngeal Weakness, Hypophrenia, Deafness, and Impaired Vision: A Novel Autosomal Dominant Myopathy with Rimmed Vacuoles

    Institute of Scientific and Technical Information of China (English)

    Ting Chen; Xiang-Hui Lu; Hui-Fang Wang; Rui Ban; Hua-Xu Liu; Qiang Shi; Qian Wang

    2016-01-01

    Background:Myopathies with rimmed vacuoles are a heterogeneous group of muscle disorders with progressive muscle weakness and varied clinical manifestations but similar features in muscle biopsies.Here,we describe a novel autosomal dominant myopathy with rimmed vacuoles in a large family with 11 patients of three generations affected.Methods:A clinical study including family history,obstetric,pediatric,and development history was recorded.Clinical examinations including physical examination,electromyography (EMG),serum creatine kinase (CK),bone X-rays,and brain magnetic resonance imaging (MRI) were performed in this family.Open muscle biopsies were performed on the proband and his mother.To find the causative gene,the whole-exome sequencing was carried out.Results:Disease onset was from adolescence to adulthood,but the affected patients of the third generation presented an earlier onset and more severe clinical manifestations than the older generations.Clinical features were characterized as dysarthria,dysphagia,external ophthalmoplegia,limb weakness,hypophrenia,deafness,and impaired vision.However,not every patient manifested all symptoms.Serum CK was mildly elevated and EMG indicated a myopathic pattern.Brain MRI showed cerebellum and brain stem mildly atrophy.Rimmed vacuoles and inclusion bodies were observed in muscle biopsy.The whole-exome sequencing was performed,but the causative gene has not been found.Conclusions:We reported a novel autosomal dominant myopathy with rimmed vacuoles characterized by dysarthria,dysphagia,external ophthalmoplegia,limb weakness,hypophrenia,deafness,and impaired vision,but the causative gene has not been found and needs further study.

  9. Determination of Glutathione and Its Redox Status in Isolated Vacuoles of Red Beetroot Cells

    Directory of Open Access Journals (Sweden)

    E.V. Pradedova

    2016-02-01

    Full Text Available The glutathione of the red beetroot vacuoles (Beta vulgaris L. was measured using three well-known methods: the spectrofluorimetric method with orthophthalic aldehyde (OPT; the spectrophotometric method with 5.5'-dithiobis-2-nitrobenzoic acid (DTNB; the high-performance liquid chromatography (HPLC. The content of reduced (GSH and oxidized glutathione (GSSG differed depending on the research method. With OPT the concentration of glutathione was: GSH – 0.059 µmol /mg protein; GSSG – 0.019 µmol/mg protein and total glutathione (GSHtotal – 0.097 µmol/mg protein. In the case of determining with DTNB the concentration of glutathione was: GSH – 0.091 µmol/mg protein; GSSG – 0.031 µmol/mg protein; GSHtotal – 0.153 µmol/mg protein. HPLC-defined concentration of glutathione was lower: GSH – 0.039 µmol/mg protein; GSSG – 0.007 µmol/mg protein; GSHtotal – 0.053 µmol/mg protein. Redox ratio of GSH/GSSG was also dependent on the method of determination: with OPT – 3.11; with DTNB – 2.96 and HPLC – 5.57. Redox ratio of glutathione in vacuoles was much lower than the tissue extracts of red beetroot, which, depending on the method of determination, was: 7.23, 7.16 and 9.22. The results showed the vacuoles of red beetroot parenchyma cells contain glutathione. Despite the low value of the redox ratio GSH/GSSG, in vacuoles the pool of reduced glutathione prevailed over the pool of oxidized glutathione.

  10. Vacuolization and apoptosis induced by nano-selenium in HeLa cell line

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    Selenium(Se),a potential drug candidate for cancer prevention,has a special property:Its nutritional dosage and tolerable upper intake level appear in a narrow range,while the therapeutic use of this mineral may depend on a higher body intake level.Nano-selenium(nano-Se) particles,however,preserve the selenium element’s low toxicity characteristic but give a high biochemical activity effect of selenium compounds.In the present study different morphologies of synthesized nano-Se were evaluated concerning its anti-proliferation and apoptosis-inducing effect.Then nano-Se(sphere) were picked out to investigate its influence on two significant events involved in apoptosis,cell cycle arrest and mitochondrial membrane potential disruption.Furthermore,massive vacuolization of HeLa cells treated by nano-Se(sphere) was observed and more methods were used to measure the level of vacuolization.Such vacuolization needs energy supply and has been demonstrated to be related to Se endocytosis.These results suggest a possible mechanism to trigger apoptosis initiation.

  11. [Effect of acetylcholine and acetylcholinesterase on the activity of contractile vacuole of Amoeba proteus].

    Science.gov (United States)

    Bagrov, Ia Iu; Manusova, N B

    2011-01-01

    Acetylcholine (ACh, 1 microM) stimulates activity of the contractile vacuole of proteus. The effect of ACh is not mimicked by its analogs which are not hydrolyzed by acetylcholinesterase (AChE), i. e., carbacholine and 5-methylfurmethide. The effect of ACh is not sensitive to the blocking action of M-cholinolytics, atropine and mytolone, but is suppressed by N-cholinolytic, tubocurarine. The inhibitors of AChE, eserine (0.01 microM) and armine (0.1 microM), suppress the effect of ACh on amoeba contractile vacuole. ACh does not affect activation of contractile vacuole induced by arginine-vasopressin (1 microM), but it blocks such effect of opiate receptors agonist, dynorphin A1-13 (0.01 microM). This effect of ACh is also suppressed by the inhibitors of AChE. These results suggest that, in the above-described effects of ACh, AChE acts not as an antagonist, but rather as a synergist.

  12. Methods to monitor autophagy in H. pylori vacuolating cytotoxin A (VacA)-treated cells.

    Science.gov (United States)

    Raju, Deepa; Jones, Nicola L

    2010-01-01

    Helicobacter pylori is a gram negative pathogen that infects at least half of the world's population and is associated not only with gastric cancer but also with other diseases such as gastritis and peptic ulcers. Indeed, H. pylori is considered the single most important risk factor for the development of gastric cancer. The vacuolating cytotoxin VacA, secreted by H. pylori, promotes intracellular survival of the bacterium and modulates host immune responses. In a recent study, we reported that VacA induces autophagy. Multilamellar autophagosomes are detected in gastric epithelial cells that are distinct from the large vacuoles formed by VacA. Furthermore, inhibition of autophagy stabilizes VacA and reduces vacuolation in the cells indicating that the toxin is being degraded by autophagy, thus limiting toxin-induced host cell damage. Many of the methods that were used for this study are commonly employed techniques that were adapted for H. pylori infection and VacA intoxication. In this paper, we describe the various methods and specific protocols used for the assessment and monitoring of autophagy during H. pylori infection.

  13. Vacuole import and degradation pathway:Insights into a specialized autophagy pathway

    Institute of Scientific and Technical Information of China (English)

    Abbas; A; Alibhoy; Hui-Ling; Chiang

    2011-01-01

    Glucose deprivation induces the synthesis of pivotagluconeogenic enzymes such as fructose-1,6-bisphos-phatase, malate dehydrogenase, phosphoenolpyruvatecarboxykinase and isocitrate lyase in Saccharomycescerevisiae. However, following glucose replenishment,these gluconeogenic enzymes are inactivated and de-graded. Studies have characterized the mechanismsby which these enzymes are inactivated in response toglucose. The site of degradation of these proteins hasalso been ascertained to be dependent on the dura-tion of starvation. Glucose replenishment of short-termstarved cells results in these proteins being degradedin the proteasome. In contrast, addition of glucose tocells starved for a prolonged period results in theseproteins being degraded in the vacuole. In the vacuoledependent pathway, these proteins are sequestered inspecialized vesicles termed vacuole import and degra-dation (Vid). These vesicles converge with the endo-cytic pathway and deliver their cargo to the vacuolefor degradation. Recent studies have identified thatinternalization, as mediated by actin polymerization, isessential for delivery of cargo proteins to the vacuolefor degradation. In addition, components of the targetof rapamycin complex 1 interact with cargo proteins during glucose starvation. Furthermore, Tor1p dissoci-ates from cargo proteins following glucose replenish-ment. Future studies will be needed to elaborate on the importance of internalization at the plasma membrane and the subsequent import of cargo proteins into Vid vesicles in the vacuole dependent degradation pathway.

  14. Mutations that affect vacuole biogenesis inhibit proliferation of the endoplasmic reticulum in Saccharomyces cerevisiae.

    Science.gov (United States)

    Koning, Ann J; Larson, Lynnelle L; Cadera, Emily J; Parrish, Mark L; Wright, Robin L

    2002-04-01

    In yeast, increased levels of the sterol biosynthetic enzyme, 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase isozyme, Hmg1p, induce assembly of nuclear-associated ER membranes called karmellae. To identify additional genes involved in karmellae assembly, we screened temperature-sensitive mutants for karmellae assembly defects. Two independently isolated, temperature-sensitive strains that were also defective for karmellae biogenesis carried mutations in VPS16, a gene involved in vacuolar protein sorting. Karmellae biogenesis was defective in all 13 other vacuole biogenesis mutants tested, although the severity of the karmellae assembly defect varied depending on the particular mutation. The hypersensitivity of 14 vacuole biogenesis mutants to tunicamycin was well correlated with pronounced defects in karmellae assembly, suggesting that the karmellae assembly defect reflected alteration of ER structure or function. Consistent with this hypothesis, seven of eight mutations causing defects in secretion also affected karmellae assembly. However, the vacuole biogenesis mutants were able to proliferate their ER in response to Hmg2p, indicating that the mutants did not have a global defect in the process of ER biogenesis.

  15. MdMYB1 Regulates Anthocyanin and Malate Accumulation by Directly Facilitating Their Transport into Vacuoles in Apples.

    Science.gov (United States)

    Hu, Da-Gang; Sun, Cui-Hui; Ma, Qi-Jun; You, Chun-Xiang; Cheng, Lailiang; Hao, Yu-Jin

    2016-03-01

    Tonoplast transporters, including proton pumps and secondary transporters, are essential for plant cell function and for quality formation of fleshy fruits and ornamentals. Vacuolar transport of anthocyanins, malate, and other metabolites is directly or indirectly dependent on the H(+)-pumping activities of vacuolar H(+)-ATPase (VHA) and/or vacuolar H(+)-pyrophosphatase, but how these proton pumps are regulated in modulating vacuolar transport is largely unknown. Here, we report a transcription factor, MdMYB1, in apples that binds to the promoters of two genes encoding the B subunits of VHA, MdVHA-B1 and MdVHA-B2, to transcriptionally activate its expression, thereby enhancing VHA activity. A series of transgenic analyses in apples demonstrates that MdMYB1/10 controls cell pH and anthocyanin accumulation partially by regulating MdVHA-B1 and MdVHA-B2. Furthermore, several other direct target genes of MdMYB10 are identified, including MdVHA-E2, MdVHP1, MdMATE-LIKE1, and MdtDT, which are involved in H(+)-pumping or in the transport of anthocyanins and malates into vacuoles. Finally, we show that the mechanism by which MYB controls malate and anthocyanin accumulation in apples also operates in Arabidopsis (Arabidopsis thaliana). These findings provide novel insights into how MYB transcription factors directly modulate the vacuolar transport system in addition to anthocyanin biosynthesis, consequently controlling organ coloration and cell pH in plants.

  16. A functional connection of Dictyostelium paracaspase with the contractile vacuole and a possible partner of the vacuolar proton ATPase

    Indian Academy of Sciences (India)

    Entsar Saheb; Ithay Biton; Katherine Maringer; John Bush

    2013-09-01

    Dictyostelium discoideum possesses only one caspase family member, paracaspase (pcp). Two separate mutant cell lines were first analysed: one cell line was an over-expressed GFP-tagged Pcp (GFP-Pcp), while the other cell line was a pcp-null (pcp-). Microscopic analysis of cells expressing GFP-Pcp revealed that Pcp was associated with the contractile vacuole membrane consisting of bladder-like vacuoles. This association was disrupted when cells were exposed to osmotic stress conditions. Compared with wild-type cells, the GFP-Pcp-over-expressing cells were susceptible to osmotic stress and were seen to be very rounded in hypo-osmotic conditions and contained more abnormally swollen contractile vacuole. Cells with pcp- were also rounded but had few, if any, contractile vacuoles. These observations suggest that Pcp is essential for Dictyostelium osmotic regulation via its functioning in the contractile vacuole system. Subjecting these cells to selected contractile vacuole inhibitor provided additional support for these findings. Furthermore, yeast two-hybrid system identified vacuolar proton ATPase (VatM) as the protein interacting with Pcp. Taken together, this work gives evidence for an eukaryotic paracaspase to be associated with both localization in and regulation of the contractile vacuolar system, an organelle critical for maintaining the normal morphology of the cell.

  17. Changes in vacuolation in the root apex cells of soybean seedlings in microgravity

    Science.gov (United States)

    Klymchuk, D. O.; Kordyum, E. L.; Vorobyova, T. V.; Chapman, D. K.; Brown, C. S.

    2003-05-01

    Changes in the vacuolation in root apex cells of soybean ( Glycine max L. [Merr.]) seedlings grown in microgravity were investigated. Spaceflight and ground control seedlings were grown in the absence or presence of KMnO 4 (to remove ethylene) for 6 days. After landing, in order to study of cell ultrastructure and subcellular free calcium ion distribution, seedling root apices were fixed in 2.5% (w/v) glutaraldehyde in 0.1 M cacodylate buffer and 2% (w/v) glutaraldehyde, 2.5% (w/v) formaldehyde, 2% (w/v) potassium antimonate K[Sb(OH) 6] in 0.1 M K 2HPO 4 buffer with an osmolarity (calculated theoretically) of 0.45 and 1.26 osmol. The concentrations of ethylene in all spaceflight canisters were significantly higher than in the ground control canisters. Seedling growth was reduced in the spaceflight-exposed plants. Additionally, the spaceflight-exposed plants exhibited progressive vacuolation in the root apex cells, particularly in the columella cells, to a greater degree than the ground controls. Plasmolysis was observed in columella cells of spaceflight roots fixed in solutions with relatively high osmolarity (1.26 osmol). The appearance of plasmolysis permitted the evaluation of the water status of cells. The water potential of the spaceflight cells was higher than the surrounding fixative solution. A decrease in osmotic potential and/or an increase in turgor potential may have induced increases in cell water potential. However, the plasmolysed (i.e. nonturgid) cells implied that increases in water potential were accompanied with a decrease in osmotic potential. In such cells changes in vacuolation may have been involved to maintain turgor pressure or may have been a result of intensification of other vacuolar functions like digestion and storage

  18. The Rice RMR1 Associates with a Distinct Prevacuolar Compartment for the Protein Storage Vacuole Pathway

    Institute of Scientific and Technical Information of China (English)

    Yun Shen; Junqi Wang; Yu Ding; SzeWan Lo; Guillaume Gouzerh; Jean-Marc Neuhaus; Liwen Jiang

    2011-01-01

    Transport of vacuolar proteins from Golgi apparatus or trans-Golgi network (TGN) to vacuoles is a receptormediated process via an intermediate membrane-bound prevacuolar compartment (PVC) in plant cells.Both vacuolar sorting receptor (VSR) and receptor homology region-transmembrane domain-RING-H2 (RMR) proteins have been shown to function in transporting storage proteins to protein storage vacuole (PSV),but little is known about the nature of the PVC for the PSV pathway.Here,we use the rice RMR1 (OsRMR1) as a probe to study the PSV pathway in plants.Immunogold electron microscopy (EM) with specific OsRMR1 antibodies showed that OsRMR1 proteins were found in the Golgi apparatus,TGN,and a distinct organelle with characteristics of PVC in both rice culture cells and developing rice seeds,as well as the protein body type Ⅱ (PBII) or PSV in developing rice seeds.This organelle,also found in both tobacco BY-2 and Arabidopsis suspension cultured cells,is morphologically distinct from the VSR-positive multivesicular lytic PVC or multivesicular body (MVB) and thus represent a PVC for the PSV pathway that we name storage PVC (sPVC).Further in vivo and in vitro interaction studies using truncated OsRMR1 proteins secreted into the culture media of transgenic BY-2 suspension cells demonstrated that OsRMR1 functions as a sorting receptor in transporting vicilin-like storage proteins.

  19. Preparative procedures markedly influence the appearance and structural integrity of protein storage vacuoles in soybean seeds.

    Science.gov (United States)

    Krishnan, Hari B

    2008-05-14

    In legumes, vacuoles serve as the final depository for storage proteins. The protein storage vacuoles (PSVs) of soybean contain electron-transparent globoid regions in which phytic acid ( myo-inositol-1,2,3,4,5,6-hexakisphosphate) is sequestered. This paper reports the effect of preparative procedures on the appearance and ultrastructural integrity of PSVs in soybeans. Electron microscopy examination of both developing and mature soybean seeds that were postfixed with osmium tetroxide revealed PSVs that had a homogeneous appearance with very few globoid crystals dispersed in them. Numerous electron-dense lipid bodies were readily seen in these cells. Omission of osmium tetroxide strikingly altered the appearance of PSVs and aided the visualization of the location of the globoids in the PSVs. In contrast to the osmicated tissue, lipid bodies appeared as electron-transparent spheres. The choice of dehydration reagent or staining procedure had little influence on the appearance of the PSVs. The results of this study demonstrate the profound effect of osmium tetroxide on the appearance and structural integrity of PSVs in soybean.

  20. Campylobacter jejuni actively invades the amoeba Acanthamoeba polyphaga and survives within non digestive vacuoles.

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    Jenny Olofsson

    Full Text Available The Gram-negative bacterium Campylobacter jejuni is able to enter, survive and multiply within the free living amoeba Acanthamoeba polyphaga, but the molecular mechanisms behind these events are still unclear. We have studied the uptake and intracellular trafficking of viable and heat killed bacterial cells of the C. jejuni strain 81-176 in A. polyphaga. We found that viable bacteria associated with a substantially higher proportion of Acanthamoeba trophozoites than heat killed bacteria. Furthermore, the kinetics of internalization, the total number of internalized bacteria as well as the intracellular localization of internalized C. jejuni were dramatically influenced by bacterial viability. Viable bacteria were internalized at a high rate already after 1 h of co-incubation and were observed in small vacuoles tightly surrounding the bacteria. In contrast, internalization of heat killed C. jejuni was low at early time points and did not peak until 96 h. These cells were gathered in large spacious vacuoles that were part of the degradative pathway as determined by the uptake of fluorescently labeled dextran. The amount of heat killed bacteria internalized by A. polyphaga did never reach the maximal amount of internalized viable bacteria. These results suggest that the uptake and intracellular survival of C. jejuni in A. polyphaga is bacterially induced.

  1. Campylobacter jejuni actively invades the amoeba Acanthamoeba polyphaga and survives within non digestive vacuoles.

    Science.gov (United States)

    Olofsson, Jenny; Axelsson-Olsson, Diana; Brudin, Lars; Olsen, Björn; Ellström, Patrik

    2013-01-01

    The Gram-negative bacterium Campylobacter jejuni is able to enter, survive and multiply within the free living amoeba Acanthamoeba polyphaga, but the molecular mechanisms behind these events are still unclear. We have studied the uptake and intracellular trafficking of viable and heat killed bacterial cells of the C. jejuni strain 81-176 in A. polyphaga. We found that viable bacteria associated with a substantially higher proportion of Acanthamoeba trophozoites than heat killed bacteria. Furthermore, the kinetics of internalization, the total number of internalized bacteria as well as the intracellular localization of internalized C. jejuni were dramatically influenced by bacterial viability. Viable bacteria were internalized at a high rate already after 1 h of co-incubation and were observed in small vacuoles tightly surrounding the bacteria. In contrast, internalization of heat killed C. jejuni was low at early time points and did not peak until 96 h. These cells were gathered in large spacious vacuoles that were part of the degradative pathway as determined by the uptake of fluorescently labeled dextran. The amount of heat killed bacteria internalized by A. polyphaga did never reach the maximal amount of internalized viable bacteria. These results suggest that the uptake and intracellular survival of C. jejuni in A. polyphaga is bacterially induced.

  2. Hitchhiking vesicular transport routes to the vacuole: amyloid recruitment to the Insoluble Protein Deposit (IPOD).

    Science.gov (United States)

    Kumar, Rajesh; Neuser, Nicole; Tyedmers, Jens

    2017-03-09

    Sequestration of aggregates into specialized deposition sites occurs in many species across all kingdoms of life ranging from bacteria to mammals and is commonly believed to have a cytoprotective function. Yeast cells possess at least three different spatially separated deposition sites, one of which is termed "Insoluble Protein Deposit (IPOD)" and harbors amyloid aggregates. We have recently discovered that recruitment of amyloid aggregates to the IPOD employs an actin cable based recruitment machinery that also involves vesicular transport (1) . Here we discuss how different proteins known to be involved in vesicular transport processes to the vacuole might act to guide amyloid aggregates to the IPOD. These factors include the Myosin V motor protein Myo2 involved in transporting vacuolar vesicles along actin cables, the transmembrane protein Atg9 involved in the recruitment of large precursor hydrolase complexes to the vacuole, the phosphatidylinositol/ phosphatidylcholine (PI/PC) transfer protein Sec 14 and the SNARE chaperone Sec 18. Furthermore, we present new data suggesting that the yeast dynamin homolog Vps1 is also crucial for faithful delivery of the amyloid model protein PrD-GFP to the IPOD. This is in agreement with a previously identified role for Vps1 in recruitment of heat-denatured aggregates to a perivacuolar deposition site (2) .

  3. Methuosis: nonapoptotic cell death associated with vacuolization of macropinosome and endosome compartments.

    Science.gov (United States)

    Maltese, William A; Overmeyer, Jean H

    2014-06-01

    Apoptosis is the most widely recognized form of physiological programmed cell death. During the past three decades, various nonapoptotic forms of cell death have gained increasing attention, largely because of their potential importance in pathological processes, toxicology, and cancer therapy. A recent addition to the panoply of cell death phenotypes is methuosis. The neologism is derived from the Greek methuo (to drink to intoxication) because the hallmark of this form of cell death is displacement of the cytoplasm by large fluid-filled vacuoles derived from macropinosomes. The demise of the cell resembles many forms of necrosis, insofar as there is a loss of metabolic capacity and plasma membrane integrity, without the cell shrinkage and nuclear fragmentation associated with apoptosis. Methuosis was initially defined in glioblastoma cells after ectopic expression of activated Ras, but recent reports have described small molecules that can induce the features of methuosis in a broad spectrum of cancer cells, including those that are resistant to conventional apoptosis-inducing drugs. This review summarizes the available information about the distinguishing morphological characteristics and underlying mechanisms of methuosis. We compare and contrast methuosis with other cytopathological conditions in which accumulation of clear cytoplasmic vacuoles is a prominent feature. Finally, we highlight key questions that need to be answered to determine whether methuosis truly represents a unique form of regulated cell death.

  4. Cdc42p and Rho1p are sequentially activated and mechanistically linked to vacuole membrane fusion

    Energy Technology Data Exchange (ETDEWEB)

    Logan, Michael R.; Jones, Lynden [Department of Cell Biology, University of Alberta, Edmonton, Alta., Canada T6G 2H7 (Canada); Eitzen, Gary, E-mail: gary.eitzen@ualberta.ca [Department of Cell Biology, University of Alberta, Edmonton, Alta., Canada T6G 2H7 (Canada)

    2010-03-26

    Small monomeric GTPases act as molecular switches, regulating many biological functions via activation of membrane localized signaling cascades. Activation of their switch function is controlled by GTP binding and hydrolysis. Two Rho GTPases, Cdc42p and Rho1p, are localized to the yeast vacuole where they regulate membrane fusion. Here, we define a method to directly examine vacuole membrane Cdc42p and Rho1p activation based on their affinity to probes derived from effectors. Cdc42p and Rho1p showed unique temporal activation which aligned with distinct subreactions of in vitro vacuole fusion. Cdc42p was rapidly activated in an ATP-independent manner while Rho1p activation was kinetically slower and required ATP. Inhibitors that are known to block vacuole membrane fusion were examined for their effect on Cdc42p and Rho1p activation. Rdi1p, which inhibits the dissociation of GDP from Rho proteins, blocked both Cdc42p and Rho1p activation. Ligands of PI(4,5)P{sub 2} specifically inhibited Rho1p activation while pre-incubation with U73122, which targets Plc1p function, increased Rho1p activation. These results define unique activation mechanisms for Cdc42p and Rho1p, which may be linked to the vacuole membrane fusion mechanism.

  5. Effect of NaCl and Helicobacter pylori vacuolating cytotoxin on cytokine expression and viability

    Institute of Scientific and Technical Information of China (English)

    Juan Sun; Kazuo Aoki; Jin-Xu Zheng; Bing-Zhong Su; Xiao-Hui Ouyang; Junichi Misumi

    2006-01-01

    AIM: To determine whether Helicobacter pylori (H pylori) vacuolating cytotoxin (VacA) regulates release of proinflammatory cytokines (IL-1β, IL-8, TNF-α, and IL-6)or alters gastric epithelial cell viability and to determine whether NaCl affects these VacA-induced changes.METHODS: Vacuolating activity was determined by measuring the uptake of neutral red into vacuoles of VacA-treated human gastric epithelial (AGS) cells. AGS cell viability was assessed by direct cell counting. Specific enzyme-linked immunosorbent assays (ELISA) and reverse transcriptase-polymerase chain reaction(RT-PCR)were performed to examine the effects of Hpylori VacA and NaCl on cell pro-inflammatory cytokine production in AGS cells. Immunohistochemical staining of gastric tissue from Mongolian gerbils was used to confirm VacA-induced pro-inflammatory cytokine production and the effects of NaCl on this VacA-induced response.RESULTS: Addition of VacA alone reduced AGS cell viability (P< 0.05), and this reduction was enhanced by high doses of NaCl (P< 0.05). VacA alone induced expression of TNF-α, IL-8 and IL-1β, while NaCl alone induced expression of TNF-α and IL-1β. Changes in mRNA levels in the presence of both VacA and NaCl were more complicated. For the case of TNF-a, expression was dosedependent on NaCl. IL-6 mRNA was not detected. However, low levels of IL-6 were detected by ELISA. Positive immunohistochemical staining of IL- 1, IL-6, and TNF-αwas found in gastric tissue of H pylori-infected gerbils fed with either a normal diet or a high salt diet. However,the staining of these three cytokines was stronger in H pylori-infected animals fed with a 5g/kg NaCl diet.CONCLUSION: VacA decreases the viability of AGS cells, and this effect can be enhanced by NaCl. NaCl also affects the production of pro-inflammatory cytokines induced by Vac A, suggesting that NaCl plays an important role in Hpylori-induced gastric epithelial cell cytotoxicity.

  6. A genome-wide immunodetection screen in S. cerevisiae uncovers novel genes involved in lysosomal vacuole function and morphology.

    Directory of Open Access Journals (Sweden)

    Florante Ricarte

    Full Text Available Vacuoles of yeast Saccharomyces cerevisiae are functionally analogous to mammalian lysosomes. Both are cellular organelles responsible for macromolecular degradation, ion/pH homeostasis, and stress survival. We hypothesized that undefined gene functions remain at post-endosomal stage of vacuolar events and performed a genome-wide screen directed at such functions at the late endosome and vacuole interface - ENV genes. The immunodetection screen was designed to identify mutants that internally accumulate precursor form of the vacuolar hydrolase carboxypeptidase Y (CPY. Here, we report the uncovering and initial characterizations of twelve ENV genes. The small size of the collection and the lack of genes previously identified with vacuolar events are suggestive of the intended exclusive functional interface of the screen. Most notably, the collection includes four novel genes ENV7, ENV9, ENV10, and ENV11, and three genes previously linked to mitochondrial processes - MAM3, PCP1, PPE1. In all env mutants, vesicular trafficking stages were undisturbed in live cells as assessed by invertase and active α-factor secretion, as well as by localization of the endocytic fluorescent marker FM4-64 to the vacuole. Several mutants exhibit defects in stress survival functions associated with vacuoles. Confocal fluorescence microscopy revealed the collection to be significantly enriched in vacuolar morphologies suggestive of fusion and fission defects. These include the unique phenotype of lumenal vesicles within vacuoles in the novel env9Δ mutant and severely fragmented vacuoles upon deletion of GET4, a gene recently implicated in tail anchored membrane protein insertion. Thus, our results establish new gene functions in vacuolar function and morphology, and suggest a link between vacuolar and mitochondrial events.

  7. Mucolipin co-deficiency causes accelerated endolysosomal vacuolation of enterocytes and failure-to-thrive from birth to weaning.

    Science.gov (United States)

    Remis, Natalie N; Wiwatpanit, Teerawat; Castiglioni, Andrew J; Flores, Emma N; Cantú, Jorge A; García-Añoveros, Jaime

    2014-12-01

    During the suckling period, intestinal enterocytes are richly endowed with endosomes and lysosomes, which they presumably utilize for the uptake and intracellular digestion of milk proteins. By weaning, mature intestinal enterocytes replace those rich in lysosomes. We found that mouse enterocytes before weaning express high levels of two endolysosomal cation channels, mucolipins 3 and 1 -products of Trpml3 and Trpml1 genes; moreover neonatal enterocytes of mice lacking both mucolipins (Trpml3-/-;Trpml1-/-) vacuolated pathologically within hours of birth and remained so until weaning. Ultrastructurally and chemically these fast-forming vacuoles resembled those that systemically appear in epithelial cells of mucolipidosis type IV (MLIV) patients, which bear mutations in Trpml1. Hence, lack of both mucolipins 1 and 3 causes an accelerated MLIV-type of vacuolation in enterocytes. The vacuoles were aberrant hybrid organelles with both endosomal and lysosomal components, and were not generated by alterations in endocytosis or exocytosis, but likely by an imbalance between fusion of lysosomes and endosomes and their subsequent scission. However, upon extensive vacuolation enterocytes displayed reduced endocytosis from the intestinal lumen, a defect expected to compromise nutrient uptake. Mice lacking both mucolipins suffered a growth delay that began after birth and continued through the suckling period but recovered after weaning, coinciding with the developmental period of enterocyte vacuolation. Our results demonstrate genetic redundancy between lysosomal mucolipins 3 and 1 in neonatal enterocytes. Furthermore, our Trpml3-/-;Trpml1-/- mice represent a polygenic animal model of the poorly-understood, and often intractable, neonatal failure-to-thrive with intestinal pathology. Our results implicate lysosomes in neonatal intestinal pathologies, a major cause of infant mortality worldwide, and suggest transient intestinal dysfunction might affect newborns with lysosomal

  8. Mucolipin co-deficiency causes accelerated endolysosomal vacuolation of enterocytes and failure-to-thrive from birth to weaning.

    Directory of Open Access Journals (Sweden)

    Natalie N Remis

    2014-12-01

    Full Text Available During the suckling period, intestinal enterocytes are richly endowed with endosomes and lysosomes, which they presumably utilize for the uptake and intracellular digestion of milk proteins. By weaning, mature intestinal enterocytes replace those rich in lysosomes. We found that mouse enterocytes before weaning express high levels of two endolysosomal cation channels, mucolipins 3 and 1 -products of Trpml3 and Trpml1 genes; moreover neonatal enterocytes of mice lacking both mucolipins (Trpml3-/-;Trpml1-/- vacuolated pathologically within hours of birth and remained so until weaning. Ultrastructurally and chemically these fast-forming vacuoles resembled those that systemically appear in epithelial cells of mucolipidosis type IV (MLIV patients, which bear mutations in Trpml1. Hence, lack of both mucolipins 1 and 3 causes an accelerated MLIV-type of vacuolation in enterocytes. The vacuoles were aberrant hybrid organelles with both endosomal and lysosomal components, and were not generated by alterations in endocytosis or exocytosis, but likely by an imbalance between fusion of lysosomes and endosomes and their subsequent scission. However, upon extensive vacuolation enterocytes displayed reduced endocytosis from the intestinal lumen, a defect expected to compromise nutrient uptake. Mice lacking both mucolipins suffered a growth delay that began after birth and continued through the suckling period but recovered after weaning, coinciding with the developmental period of enterocyte vacuolation. Our results demonstrate genetic redundancy between lysosomal mucolipins 3 and 1 in neonatal enterocytes. Furthermore, our Trpml3-/-;Trpml1-/- mice represent a polygenic animal model of the poorly-understood, and often intractable, neonatal failure-to-thrive with intestinal pathology. Our results implicate lysosomes in neonatal intestinal pathologies, a major cause of infant mortality worldwide, and suggest transient intestinal dysfunction might affect newborns

  9. An NPF transporter exports a central monoterpene indole alkaloid intermediate from the vacuole.

    Science.gov (United States)

    Payne, Richard M E; Xu, Deyang; Foureau, Emilien; Teto Carqueijeiro, Marta Ines Soares; Oudin, Audrey; Bernonville, Thomas Dugé de; Novak, Vlastimil; Burow, Meike; Olsen, Carl-Erik; Jones, D Marc; Tatsis, Evangelos C; Pendle, Ali; Ann Halkier, Barbara; Geu-Flores, Fernando; Courdavault, Vincent; Nour-Eldin, Hussam Hassan; O'Connor, Sarah E

    2017-01-13

    Plants sequester intermediates of metabolic pathways into different cellular compartments, but the mechanisms by which these molecules are transported remain poorly understood. Monoterpene indole alkaloids, a class of specialized metabolites that includes the anticancer agent vincristine, antimalarial quinine and neurotoxin strychnine, are synthesized in several different cellular locations. However, the transporters that control the movement of these biosynthetic intermediates within cellular compartments have not been discovered. Here we present the discovery of a tonoplast localized nitrate/peptide family (NPF) transporter from Catharanthus roseus, CrNPF2.9, that exports strictosidine, the central intermediate of this pathway, into the cytosol from the vacuole. This discovery highlights the role that intracellular localization plays in specialized metabolism, and sets the stage for understanding and controlling the central branch point of this pharmacologically important group of compounds.

  10. Coxiella burnetii and Leishmania mexicana residing within similar parasitophorous vacuoles elicit disparate host responses

    Directory of Open Access Journals (Sweden)

    Jess A Millar

    2015-08-01

    Full Text Available Coxiella burnetii is a bacterium that thrives in an acidic parasitophorous vacuole (PV derived from lysosomes. Leishmania mexicana, a eukaryote, has also independently evolved to live in a morphologically similar PV. As Coxiella and Leishmania are highly divergent organisms that cause different diseases, we reasoned that their respective infections would likely elicit distinct host responses despite producing phenotypically similar parasite-containing vacuoles. The objective of this study was to investigate, at the molecular level, the macrophage response to each pathogen. Infection of THP-1 (human monocyte/macrophage cells with Coxiella and Leishmania elicited disparate host responses. At 5 days post-infection, when compared to uninfected cells, 1057 genes were differentially expressed (746 genes up- and 311 genes down-regulated in C. burnetii infected cells, whereas 698 genes (534 genes up- and 164 genes down-regulated were differentially expressed in L. mexicana infected cells. Interestingly, of the 1755 differentially expressed genes identified in this study, only 126 genes (~7% are common to both infections. We also discovered that 1090 genes produced mRNA isoforms at significantly different levels under the two infection conditions, suggesting that alternate proteins encoded by the same gene might have important roles in host response to each infection. Additionally, we detected 257 micro RNAs (miRNAs that were expressed in THP-1 cells and identified miRNAs that were specifically expressed during Coxiella or Leishmania infections. Collectively, this study identified host mRNAs and miRNAs that were influenced by Coxiella and/or Leishmania infections. Intriguingly, our data indicate that although their PVs are morphologically similar, Coxiella and Leishmania have evolved different strategies that perturb distinct host processes to create and thrive within their respective intracellular niches.

  11. Determinants of GBP recruitment to Toxoplasma gondii vacuoles and the parasitic factors that control it.

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    Sebastian Virreira Winter

    Full Text Available IFN-γ is a major cytokine that mediates resistance against the intracellular parasite Toxoplasma gondii. The p65 guanylate-binding proteins (GBPs are strongly induced by IFN-γ. We studied the behavior of murine GBP1 (mGBP1 upon infection with T. gondii in vitro and confirmed that IFN-γ-dependent re-localization of mGBP1 to the parasitophorous vacuole (PV correlates with the virulence type of the parasite. We identified three parasitic factors, ROP16, ROP18, and GRA15 that determine strain-specific accumulation of mGBP1 on the PV. These highly polymorphic proteins are held responsible for a large part of the strain-specific differences in virulence. Therefore, our data suggest that virulence of T. gondii in animals may rely in part on recognition by GBPs. However, phagosomes or vacuoles containing Trypanosoma cruzi did not recruit mGBP1. Co-immunoprecipitation revealed mGBP2, mGBP4, and mGBP5 as binding partners of mGBP1. Indeed, mGBP2 and mGBP5 co-localize with mGBP1 in T. gondii-infected cells. T. gondii thus elicits a cell-autonomous immune response in mice with GBPs involved. Three parasitic virulence factors and unknown IFN-γ-dependent host factors regulate this complex process. Depending on the virulence of the strains involved, numerous GBPs are brought to the PV as part of a large, multimeric structure to combat T. gondii.

  12. Food vacuole associated enolase in plasmodium undergoes multiple post-translational modifications: evidence for atypical ubiquitination.

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    Saudamini Shevade

    Full Text Available Plasmodium enolase localizes to several sub-cellular compartments viz. cytosol, nucleus, cell membrane, food vacuole (FV and cytoskeleton, without having any organelle targeting signal sequences. This enzyme has been shown to undergo multiple post-translational modifications (PTMs giving rise to several variants that show organelle specific localization. It is likely that these PTMs may be responsible for its diverse distribution and moonlighting functions. While most variants have a MW of ~50 kDa and are likely to arise due to changes in pI, food vacuole (FV associated enolase showed three forms with MW~50, 65 and 75 kDa. Evidence from immuno-precipitation and western analysis indicates that the 65 and 75 kDa forms of FV associated enolase are ubiquitinated. Using mass spectrometry (MS, definitive evidence is obtained for the nature of PTMs in FV associated variants of enolase. Results showed several modifications, viz. ubiquitination at K147, phosphorylation at Y148 and acetylation at K142 and K384. MS data also revealed the conjugation of three ubiquitin (Ub molecules to enolase through K147. Trimeric ubiquitin has a linear peptide linkage between the NH2-terminal methionine of the first ubiquitin (Ub1 and the C-terminal G76 of the second (Ub2. Ub2 and third ubiquitin (Ub3 were linked through an atypical isopeptide linkage between K6 of Ub2 and G76 of Ub3, respectively. Further, the tri-ubiquitinated form was found to be largely associated with hemozoin while the 50 and 65 kDa forms were present in the NP-40 soluble fraction of FV. Mass spectrometry results also showed phosphorylation of S42 in the cytosolic enolase from P. falciparum and T337 in the cytoskeleton associated enolase from P. yoelii. The composition of food vacuolar proteome and likely interactors of enolase are also being reported.

  13. Hyperacidification of Vacuoles by the Combined Action of Two Different P-ATPases in the Tonoplast Determines Flower Color

    Directory of Open Access Journals (Sweden)

    Marianna Faraco

    2014-01-01

    Full Text Available The acidification of endomembrane compartments is essential for enzyme activities, sorting, trafficking, and trans-membrane transport of various compounds. Vacuoles are mildly acidic in most plant cells because of the action of V-ATPase and/or pyrophosphatase proton pumps but are hyperacidified in specific cells by mechanisms that remained unclear. Here, we show that the blue petal color of petunia ph mutants is due to a failure to hyperacidify vacuoles. We report that PH1 encodes a P3B-ATPase, hitherto known as Mg2+ transporters in bacteria only, that resides in the vacuolar membrane (tonoplast. In vivo nuclear magnetic resonance and genetic data show that PH1 is required and, together with the tonoplast H+ P3A-ATPase PH5, sufficient to hyperacidify vacuoles. PH1 has no H+ transport activity on its own but can physically interact with PH5 and boost PH5 H+ transport activity. Hence, the hyperacidification of vacuoles in petals, and possibly other tissues, relies on a heteromeric P-ATPase pump.

  14. The vacuolating cytotoxin of Helicobacter pylori%幽门螺杆菌空泡毒素研究进展

    Institute of Scientific and Technical Information of China (English)

    刘纯杰; 陶好霞; 张兆山

    2001-01-01

    幽门螺杆菌空泡毒素是该菌产生的与已知其它细菌毒素无明显同源性的唯一蛋白毒素。该毒素是幽门螺杆菌重要的毒力致病因子,它的产生与感染者胃肠上皮损伤和溃疡形成密切相关。本文就幽门螺杆菌空泡毒素的结构与功能研究进展以及在未来免疫预防与免疫治疗中的作用进行了简述。%The vacuolating cytotoxin is a unique proteinous cytotoxin producted by H. pylori that showed no striking primary sequence homology with other known baterial toxins. The cytotoxin is an important fator in the pathogenesis of H. pylori, which induces vacuolation of epithelial cells and plays an important role in gastric epithelial necrosis and peptic ulceration. In the paper, the progress on structure and function of the vacuolating cytotoxin of H. pylori and the roles of the H. pylori vacuolating cytotoxin in the future immunoprophylaxis and immunotherapy were reviewed.

  15. Quantitative trait loci affecting phenotypic variation in the vacuolated lens mouse mutant, a multigenic mouse model of neural tube defects

    NARCIS (Netherlands)

    Korstanje, Ron; Desai, Jigar; Lazar, Gloria; King, Benjamin; Rollins, Jarod; Spurr, Melissa; Joseph, Jamie; Kadambi, Sindhuja; Li, Yang; Cherry, Allison; Matteson, Paul G.; Paigen, Beverly; Millonig, James H.

    2008-01-01

    Korstanje R, Desai J, Lazar G, King B, Rollins J, Spurr M, Joseph J, Kadambi S, Li Y, Cherry A, Matteson PG, Paigen B, Millonig JH. Quantitative trait loci affecting phenotypic variation in the vacuolated lens mouse mutant, a multigenic mouse model of neural tube defects. Physiol Genomics 35: 296-30

  16. LAMP proteins account for the maturation delay during the establishment of the Coxiella burnetii-containing vacuole.

    Science.gov (United States)

    Schulze-Luehrmann, Jan; Eckart, Rita A; Ölke, Martha; Saftig, Paul; Liebler-Tenorio, Elisabeth; Lührmann, Anja

    2016-02-01

    The obligate intracellular pathogen Coxiella burnetii replicates in a large phagolysosomal-like vacuole. Currently, both host and bacterial factors required for creating this replicative parasitophorous C. burnetii-containing vacuole (PV) are poorly defined. Here, we assessed the contributions of the most abundant proteins of the lysosomal membrane, LAMP-1 and LAMP-2, to the establishment and maintenance of the PV. Whereas these proteins were not critical for uptake of C. burnetii, they influenced the intracellular replication of C. burnetii. In LAMP-1/2 double-deficient fibroblasts as well as in LAMP-1/2 knock-down cells, C. burnetii establishes a significantly smaller, yet faster maturing vacuole, which harboured more bacteria. The accelerated maturation of PVs in LAMP double-deficient fibroblasts, which was partially or fully reversed by ectopic expression of LAMP-1 or LAMP-2, respectively, was characterized by an increased fusion rate with endosomes, lysosomes and bead-containing phagosomes, but not by different fusion kinetics with autophagy vesicles. These findings establish that LAMP proteins are critical for the maturation delay of PVs. Unexpectedly, neither the creation of the spacious vacuole nor the delay in maturation was found to be prerequisites for the intracellular replication of C. burnetii.

  17. Emergence of the Terrestrial Ciliate Colpoda cucullus from a Resting Cyst: Rupture of the Cyst Wall by Active Expansion of an Excystment Vacuole

    Science.gov (United States)

    Funadani, Ryoji; Suetomo, Yasutaka; Matsuoka, Tatsuomi

    2013-01-01

    The first sign of excysting Colpoda cucullus cells is the initiation of the pulsation of a contractile vacuole, which is then replaced by a non-pulsating vacuole (excystment vacuole) that continues to expand and finally ruptures the outermost cyst wall (ectocyst) due to inner pressure. A ciliate surrounded by flexible membranes (endocyst) thus emerges. The osmolarity of the excysting cells is estimated to be 140 mOsm L−1 from the relationship between the frequency of contractile vacuole pulsation and the external sucrose concentration. Both the expansion of the excystment vacuole and the emergence of ciliates occurred even when the cysts were immersed in hypertonic medium. In hypotonic medium containing sodium azide (NaN3, a cytochrome c oxidase inhibitor), the contractile vacuole of vegetative cells stopped pulsating and gradually expanded, causing cells to burst. When C. cucullus was induced to encyst in a hypotonic medium containing NaN3, the expansion of the excystment vacuoles was inhibited. These results suggest that the active uptake of water may be responsible for the expansion of the excystment vacuole required for the ectocyst to rupture. PMID:23268793

  18. Poorly differentiated angiosarcoma without vasoformative channels but with focal intracytoplastic vacuoles mimicking liposarcomas

    Directory of Open Access Journals (Sweden)

    Tadashi Terada, MD, PhD

    2016-03-01

    Full Text Available Angiosarcoma (AS showed diverse morphologies from well formed malignant vasculatures to poorly differentiated tumor with only a few clues of endothelial differentiation. Herein reported are two cases of AS without primitive vasoformative channels (VC. They showed, instead, a very few foci of intracytoplasmic vacuoles (ICV that mimicked liposarcoma. The two cases were found in 12 cases of AS in computer database. Both are men, 57 and 68 years. One is cutaneous (foot AS and another is soft tissue (thigh AS. The largest diameter of cutaneous AS was 5 cm, and that of soft tissue AS 9 cm. The prognosis of both patients was poor; both died of metastases 4 and 6 years after initial presentation. In both cases, hematoxylin and eosin (HE diagnosis was difficult because there were no VC, and most of the tumors were composed of primitive mesenchymal tissues. In both cases, however, a few very tiny foci consisting of ICV were seen. At first, the author considered them as mucins or fat, and suspected liposarcoma. In fact, they were pseudolipoblasts. Several mucin stains showed no mucins, and fat stains of frozen sections of formalin fixed tissue were negative for fat. Immunohistochemically, the vacuoles were positive for factor VIII-related antigen (F-VIII-RA, Ulex lectin, CD31, CD34, vimentin, p53 and Ki-67 (labeling index = 64% and 75%, but negative for various types of cytokeratins (CK, EMA, CEA, CA19-9, CD45, smooth muscle actins, S100 protein, myoglobin, HMB-45, Melan A, NCAM, and NSE. F-VIII-RA is specific and Ulex lectin and CD31 are relatively specific for endothelium. Therefore, the pathological diagnosis of AS could be made by the combined histologic features (ICV and Immunohistochemical positivity of F-VIII-RA, Ulex lectin, and CD31. Thus, it appeared that the ICV may be the only clue of poorly differentiated or undifferentiated AS. In such undifferentiated cases, combined observations of meticulous histologic observations (intracytoplasmic

  19. Mitochondrial targeted β-lapachone induces mitochondrial dysfunction and catastrophic vacuolization in cancer cells.

    Science.gov (United States)

    Ma, Jing; Lim, Chaemin; Sacher, Joshua R; Van Houten, Bennett; Qian, Wei; Wipf, Peter

    2015-11-01

    Mitochondria play important roles in tumor cell physiology and survival by providing energy and metabolites for proliferation and metastasis. As part of their oncogenic status, cancer cells frequently produce increased levels of mitochondrial-generated reactive oxygen species (ROS). However, extensive stimulation of ROS generation in mitochondria has been shown to be able to induce cancer cell death, and is one of the major mechanisms of action of many anticancer agents. We hypothesized that enhancing mitochondrial ROS generation through direct targeting of a ROS generator into mitochondria will exhibit tumor cell selectivity, as well as high efficacy in inducing cancer cell death. We thus synthesized a mitochondrial targeted version of β-lapachone (XJB-Lapachone) based on our XJB mitochondrial targeting platform. We found that the mitochondrial targeted β-lapachone is more efficient in inducing apoptosis compared to unconjugated β-lapachone, and the tumor cell selectivity is maintained. XJB-Lapachone also induced extensive cellular vacuolization and autophagy at a concentration not observed with unconjugated β-lapachone. Through characterization of mitochondrial function we revealed that XJB-Lapachone is indeed more capable of stimulating ROS generation in mitochondria, which led to a dramatic mitochondrial uncoupling and autophagic degradation of mitochondria. Taken together, we have demonstrated that targeting β-lapachone accomplishes higher efficacy through inducing ROS generation directly in mitochondria, resulting in extensive mitochondrial and cellular damage. XJB-Lapachone will thus help to establish a novel platform for the design of next generation mitochondrial targeted ROS generators for cancer therapy.

  20. Identification of vacuoles containing extraintestinal differentiated forms of Legionella pneumophila in colonized Caenorhabditis elegans soil nematodes.

    Science.gov (United States)

    Hellinga, Jacqueline R; Garduño, Rafael A; Kormish, Jay D; Tanner, Jennifer R; Khan, Deirdre; Buchko, Kristyn; Jimenez, Celine; Pinette, Mathieu M; Brassinga, Ann Karen C

    2015-08-01

    Legionella pneumophila, a causative agent of Legionnaires' disease, is a facultative intracellular parasite of freshwater protozoa. Legionella pneumophila features a unique developmental network that involves several developmental forms including the infectious cyst forms. Reservoirs of L. pneumophila include natural and man-made freshwater systems; however, recent studies have shown that isolates of L. pneumophila can also be obtained directly from garden potting soil suggesting the presence of an additional reservoir. A previous study employing the metazoan Caenorhabditis elegans, a member of the Rhabditidae family of free-living soil nematodes, demonstrated that the intestinal lumen can be colonized with L. pneumophila. While both replicative forms and differentiated forms were observed in C. elegans, these morphologically distinct forms were initially observed to be restricted to the intestinal lumen. Using live DIC imaging coupled with focused transmission electron microscopy analyses, we report here that L. pneumophila is able to invade and establish Legionella-containing vacuoles (LCVs) in the intestinal cells. In addition, LCVs containing replicative and differentiated cyst forms were observed in the pseudocoelomic cavity and gonadal tissue of nematodes colonized with L. pneumophila. Furthermore, establishment of LCVs in the gonadal tissue was Dot/Icm dependent and required the presence of the endocytic factor RME-1 to gain access to maturing oocytes. Our findings are novel as this is the first report, to our knowledge, of extraintestinal LCVs containing L. pneumophila cyst forms in C. elegans tissues, highlighting the potential of soil-dwelling nematodes as an alternate environmental reservoir for L. pneumophila.

  1. New Challenges for the Design of High Value Plant Products: Stabilization of Anthocyanins in Plant Vacuoles.

    Science.gov (United States)

    Passeri, Valentina; Koes, Ronald; Quattrocchio, Francesca M

    2016-01-01

    In the last decade plant biotechnologists and breeders have made several attempt to improve the antioxidant content of plant-derived food. Most efforts concentrated on increasing the synthesis of antioxidants, in particular anthocyanins, by inducing the transcription of genes encoding the synthesizing enzymes. We present here an overview of economically interesting plant species, both food crops and ornamentals, in which anthocyanin content was improved by traditional breeding or transgenesis. Old genetic studies in petunia and more recent biochemical work in brunfelsia, have shown that after synthesis and compartmentalization in the vacuole, anthocyanins need to be stabilized to preserve the color of the plant tissue over time. The final yield of antioxidant molecules is the result of the balance between synthesis and degradation. Therefore the understanding of the mechanism that determine molecule stabilization in the vacuolar lumen is the next step that needs to be taken to further improve the anthocyanin content in food. In several species a phenomenon known as fading is responsible for the disappearance of pigmentation which in some case can be nearly complete. We discuss the present knowledge about the genetic and biochemical factors involved in pigment preservation/destabilization in plant cells. The improvement of our understanding of the fading process will supply new tools for both biotechnological approaches and marker-assisted breeding.

  2. Transport of Arginine and Aspartic Acid into Isolated Barley Mesophyll Vacuoles 1

    Science.gov (United States)

    Martinoia, Enrico; Thume, Monika; Vogt, Esther; Rentsch, Doris; Dietz, Karl-Josef

    1991-01-01

    The transport of arginine into isolated barley (Hordeum vulgare L.) mesophyll vacuoles was investigated. In the absence of ATP, arginine uptake was saturable with a Km of 0.3 to 0.4 millimolar. Positively charged amino acids inhibited arginine uptake, lysine being most potent with a Ki of 1.2 millimolar. In the presence of free ATP, but not of its Mg-complex, uptake of arginine was drastically enhanced and a linear function of its concentration up to 16 millimolar. The nonhydrolyzable adenylyl imidodiphosphate, but no other nucleotide tested, could substitute for ATP. Therefore, it is suggested that this process does not require energy and does not involve the tonoplast ATPase. The ATP-dependent arginine uptake was strongly inhibited by p-chloromercuriphenylsulfonic acid. Furthermore, hydrophobic amino acids were inhibitory (I50 phenylalanine 1 millimolar). Similar characteristics were observed for the uptake of aspartic acid. However, rates of ATP-stimulated aspartic acid transport were 10-fold lower as compared to arginine transport. Uptake of aspartate in the absence of ATP was negligible. PMID:16668447

  3. Structure of CARDS toxin, a unique ADP-ribosylating and vacuolating cytotoxin from Mycoplasma pneumoniae.

    Science.gov (United States)

    Becker, Argentina; Kannan, T R; Taylor, Alexander B; Pakhomova, Olga N; Zhang, Yanfeng; Somarajan, Sudha R; Galaleldeen, Ahmad; Holloway, Stephen P; Baseman, Joel B; Hart, P John

    2015-04-21

    Mycoplasma pneumoniae (Mp) infections cause tracheobronchitis and "walking" pneumonia, and are linked to asthma and other reactive airway diseases. As part of the infectious process, the bacterium expresses a 591-aa virulence factor with both mono-ADP ribosyltransferase (mART) and vacuolating activities known as Community-Acquired Respiratory Distress Syndrome Toxin (CARDS TX). CARDS TX binds to human surfactant protein A and annexin A2 on airway epithelial cells and is internalized, leading to a range of pathogenetic events. Here we present the structure of CARDS TX, a triangular molecule in which N-terminal mART and C-terminal tandem β-trefoil domains associate to form an overall architecture distinct from other well-recognized ADP-ribosylating bacterial toxins. We demonstrate that CARDS TX binds phosphatidylcholine and sphingomyelin specifically over other membrane lipids, and that cell surface binding and internalization activities are housed within the C-terminal β-trefoil domain. The results enhance our understanding of Mp pathogenicity and suggest a novel avenue for the development of therapies to treat Mp-associated asthma and other acute and chronic airway diseases.

  4. The diverse and dynamic nature of Leishmania parasitophorous vacuoles studied by multidimensional imaging.

    Directory of Open Access Journals (Sweden)

    Fernando Real

    Full Text Available An important area in the cell biology of intracellular parasitism is the customization of parasitophorous vacuoles (PVs by prokaryotic or eukaryotic intracellular microorganisms. We were curious to compare PV biogenesis in primary mouse bone marrow-derived macrophages exposed to carefully prepared amastigotes of either Leishmania major or L. amazonensis. While tight-fitting PVs are housing one or two L. major amastigotes, giant PVs are housing many L. amazonensis amastigotes. In this study, using multidimensional imaging of live cells, we compare and characterize the PV biogenesis/remodeling of macrophages i hosting amastigotes of either L. major or L. amazonensis and ii loaded with Lysotracker, a lysosomotropic fluorescent probe. Three dynamic features of Leishmania amastigote-hosting PVs are documented: they range from i entry of Lysotracker transients within tight-fitting, fission-prone L. major amastigote-housing PVs; ii the decrease in the number of macrophage acidic vesicles during the L. major PV fission or L. amazonensis PV enlargement; to iii the L. amazonensis PV remodeling after homotypic fusion. The high content information of multidimensional images allowed the updating of our understanding of the Leishmania species-specific differences in PV biogenesis/remodeling and could be useful for the study of other intracellular microorganisms.

  5. New challenges for the design of high value plant products: stabilization of anthocyanins in plant vacuoles

    Directory of Open Access Journals (Sweden)

    Valentina ePasseri

    2016-02-01

    Full Text Available In the last decade plant biotechnologists and breeders have made several attempt to improve the antioxidant content of plant-derived food. Most efforts concentrated on increasing the synthesis of antioxidants, in particular anthocyanins, by inducing the transcription of genes encoding the synthesizing enzymes. We present here an overview of economically interesting plant species, both food crops and ornamentals, in which anthocyanin content was improved by traditional breeding or transgenesis. Old genetic studies in petunia and more recent biochemical work in brunfelsia, have shown that after synthesis and compartmentalization in the vacuole, anthocyanins need to be stabilized to preserve the color of the plant tissue over time. The final yield of antioxidant molecules is the result of the balance between synthesis and degradation. Therefore the understanding of the mechanism that determine molecule stabilization in the vacuolar lumen is the next step that needs to be taken to further improve the anthocyanin content in food.In several species a phenomenon known as fading is responsible for the disappearance of pigmentation which in some case can be nearly complete. We discuss the present knowledge about the genetic and biochemical factors involved in pigment preservation/destabilization in plant cells.The improvement of our understanding of the fading process will supply new tools for both biotechnological approaches and marker-assisted breeding.

  6. Delivery of prolamins to the protein storage vacuole in maize aleurone cells.

    Science.gov (United States)

    Reyes, Francisca C; Chung, Taijoon; Holding, David; Jung, Rudolf; Vierstra, Richard; Otegui, Marisa S

    2011-02-01

    Zeins, the prolamin storage proteins found in maize (Zea mays), accumulate in accretions called protein bodies inside the endoplasmic reticulum (ER) of starchy endosperm cells. We found that genes encoding zeins, α-globulin, and legumin-1 are transcribed not only in the starchy endosperm but also in aleurone cells. Unlike the starchy endosperm, aleurone cells accumulate these storage proteins inside protein storage vacuoles (PSVs) instead of the ER. Aleurone PSVs contain zein-rich protein inclusions, a matrix, and a large system of intravacuolar membranes. After being assembled in the ER, zeins are delivered to the aleurone PSVs in atypical prevacuolar compartments that seem to arise at least partially by autophagy and consist of multilayered membranes and engulfed cytoplasmic material. The zein-containing prevacuolar compartments are neither surrounded by a double membrane nor decorated by AUTOPHAGY RELATED8 protein, suggesting that they are not typical autophagosomes. The PSV matrix contains glycoproteins that are trafficked through a Golgi-multivesicular body (MVB) pathway. MVBs likely fuse with the multilayered, autophagic compartments before merging with the PSV. The presence of similar PSVs also containing prolamins and large systems of intravacuolar membranes in wheat (Triticum aestivum) and barley (Hordeum vulgare) starchy endosperm suggests that this trafficking mechanism may be common among cereals.

  7. Expression of 87 kD protein in the broth culture filtrate of Helicobacter pylori and its association with the vacuolating effect

    Institute of Scientific and Technical Information of China (English)

    SHI Li; YIE Gui-an; NAN Qing-zhen; SUN Yong; ZHANG Ya-li; ZHANG Zhen-shu; ZHOU Dian-yuan

    2001-01-01

    To study the vacuolating effect of Helicobacter pylori(H.pylori). Method: The vacuolating effect and its relationship with vacuolating cytotoxin antigen (an 87 kD protein) were investigated by the method of cytotoxic test, SDS-PAGE and scanning. Result: Of the 62 clinical isolates, 43 strains were H.pylori (Toxin+) with vacuolating effect, while the others were H.pylori (Toxin-) without vacuolating effect. Altogether 78.26%(36/46) patients with peptic ulcer were infected with H.pylori (Toxin+) strains, and only 42.86%(6/14) who had gastritis were infected with H.pylori (Toxin+) strains, with significant difference between them(χ2=4.83,P<0.05). A protein with relativemolecular mass of 87 kD was identified in the broth culture filter(BCF) of 30.23% H. Pylori (Toxin+) strains (13/43) but in none of that of H.pylori (Toxin-) strains, and the difference was statistically significant(P<0.05). There was a significant and concordant relationship between the OD value of the protein band and the titer of vacuolating activity of H.pylori (Toxin+) (r=0.67 and P<0.05 by linear regression analysis). Conclusion: H.pylori (Toxin+) were more often associated with peptic ulcerous diseases than with gastritis diseases. The vacuolating effect of H.pylori (Toxin+) may be caused by the 87 kD protein.

  8. Fluorescence imaging analysis of taxol-induced ASTC-a-1 cell death with cell swelling and cytoplasmic vacuolization

    Science.gov (United States)

    Chen, Tong-sheng; Sun, Lei; Wang, Longxiang; Wang, Huiying

    2008-02-01

    Taxol (Paclitaxel), an isolated component from the bark of the Pacific yew Taxus brevifolia, exhibits a broad spectrum of clinical activity against human cancers. Taxol can promote microtubule (MT) assembly, inhibit depolymerization, and change MT dynamics, resulting in disruption of the normal reorganization of the microtubule network required for mitosis and cell proliferation. However, the molecular mechanism of taxol-induced cell death is still unclear. In this report, CCK-8 was used to assay the inhibition of taxol on the human lung adenocarcinoma (ASTC-a-1) cells viability, confocal fluorescence microscope was used to monitor the morphology changes of cells with taxol treatment. We for the first time describe the characteristics of taxol-induced cells swelling, cytoplasmic vacuolization and cell death. Taxol induced swelling, cytoplasmatic vacuolization and cell death without cell shrinkage and membrane rupture. These features differ from those of apoptosis and resemble the paraptosis, a novel nonapoptotic PCD.

  9. In Vivo Biotinylation of the Toxoplasma Parasitophorous Vacuole Reveals Novel Dense Granule Proteins Important for Parasite Growth and Pathogenesis

    Science.gov (United States)

    Nadipuram, Santhosh M.; Kim, Elliot W.; Vashisht, Ajay A.; Lin, Andrew H.; Bell, Hannah N.; Coppens, Isabelle; Wohlschlegel, James A.

    2016-01-01

    ABSTRACT Toxoplasma gondii is an obligate intracellular parasite that invades host cells and replicates within a unique parasitophorous vacuole. To maintain this intracellular niche, the parasite secretes an array of dense granule proteins (GRAs) into the nascent parasitophorous vacuole. These GRAs are believed to play key roles in vacuolar remodeling, nutrient uptake, and immune evasion while the parasite is replicating within the host cell. Despite the central role of GRAs in the Toxoplasma life cycle, only a subset of these proteins have been identified, and many of their roles have not been fully elucidated. In this report, we utilize the promiscuous biotin ligase BirA* to biotinylate GRA proteins secreted into the vacuole and then identify those proteins by affinity purification and mass spectrometry. Using GRA-BirA* fusion proteins as bait, we have identified a large number of known and candidate GRAs and verified localization of 13 novel GRA proteins by endogenous gene tagging. We proceeded to functionally characterize three related GRAs from this group (GRA38, GRA39, and GRA40) by gene knockout. While Δgra38 and Δgra40 parasites showed no altered phenotype, disruption of GRA39 results in slow-growing parasites that contain striking lipid deposits in the parasitophorous vacuole, suggesting a role in lipid regulation that is important for parasite growth. In addition, parasites lacking GRA39 showed dramatically reduced virulence and a lower tissue cyst burden in vivo. Together, the findings from this work reveal a partial vacuolar proteome of T. gondii and identify a novel GRA that plays a key role in parasite replication and pathogenesis. PMID:27486190

  10. A Legionella pneumophila effector protein encoded in a region of genomic plasticity binds to Dot/Icm-modified vacuoles.

    Directory of Open Access Journals (Sweden)

    Shira Ninio

    2009-01-01

    Full Text Available Legionella pneumophila is an opportunistic pathogen that can cause a severe pneumonia called Legionnaires' disease. In the environment, L. pneumophila is found in fresh water reservoirs in a large spectrum of environmental conditions, where the bacteria are able to replicate within a variety of protozoan hosts. To survive within eukaryotic cells, L. pneumophila require a type IV secretion system, designated Dot/Icm, that delivers bacterial effector proteins into the host cell cytoplasm. In recent years, a number of Dot/Icm substrate proteins have been identified; however, the function of most of these proteins remains unknown, and it is unclear why the bacterium maintains such a large repertoire of effectors to promote its survival. Here we investigate a region of the L. pneumophila chromosome that displays a high degree of plasticity among four sequenced L. pneumophila strains. Analysis of GC content suggests that several genes encoded in this region were acquired through horizontal gene transfer. Protein translocation studies establish that this region of genomic plasticity encodes for multiple Dot/Icm effectors. Ectopic expression studies in mammalian cells indicate that one of these substrates, a protein called PieA, has unique effector activities. PieA is an effector that can alter lysosome morphology and associates specifically with vacuoles that support L. pneumophila replication. It was determined that the association of PieA with vacuoles containing L. pneumophila requires modifications to the vacuole mediated by other Dot/Icm effectors. Thus, the localization properties of PieA reveal that the Dot/Icm system has the ability to spatially and temporally control the association of an effector with vacuoles containing L. pneumophila through activities mediated by other effector proteins.

  11. The Formation of Anthocyanic Vacuolar Inclusions in Arabidopsis thaliana and Implications for the Sequestration of Anthocyanin Pigments

    OpenAIRE

    2009-01-01

    Anthocyanins are flavonoid pigments that accumulate in the large central vacuole of most plants. Inside the vacuole, anthocyanins can be found uniformly distributed or as part of sub-vacuolar pigment bodies, the Anthocyanic Vacuolar Inclusions (AVIs). Using Arabidopsis seedlings grown under anthocyanin-inductive conditions as a model to understand how AVIs are formed, we show here that the accumulation of AVIs strongly correlates with the formation of cyanidin 3-glucoside (C3G) and derivative...

  12. The protein transportation pathway from Golgi to vacuoles via endosomes plays a role in enhancement of methylmercury toxicity

    Science.gov (United States)

    Hwang, Gi-Wook; Murai, Yasutaka; Takahashi, Tsutomu; Naganuma, Akira

    2014-07-01

    Methylmercury causes serious damage to the central nervous system, but the molecular mechanisms of methylmercury toxicity are only marginally understood. In this study, we used a gene-deletion mutant library of budding yeast to conduct genome-wide screening for gene knockouts affecting the sensitivity of methylmercury toxicity. We successfully identified 31 genes whose deletions confer resistance to methylmercury in yeast, and 18 genes whose deletions confer hypersensitivity to methylmercury. Yeast genes whose deletions conferred resistance to methylmercury included many gene encoding factors involved in protein transport to vacuoles. Detailed examination of the relationship between the factors involved in this transport system and methylmercury toxicity revealed that mutants with loss of the factors involved in the transportation pathway from the trans-Golgi network (TGN) to the endosome, protein uptake into the endosome, and endosome-vacuole fusion showed higher methylmercury resistance than did wild-type yeast. The results of our genetic engineering study suggest that this vesicle transport system (proteins moving from the TGN to vacuole via endosome) is responsible for enhancing methylmercury toxicity due to the interrelationship between the pathways. There is a possibility that there may be proteins in the cell that enhance methylmercury toxicity through the protein transport system.

  13. Sat, the secreted autotransporter toxin of uropathogenic Escherichia coli, is a vacuolating cytotoxin for bladder and kidney epithelial cells.

    Science.gov (United States)

    Guyer, Debra M; Radulovic, Suzana; Jones, Faye-Ellen; Mobley, Harry L T

    2002-08-01

    The secreted autotransporter toxin (Sat) of uropathogenic Escherichia coli exhibits cytopathic activity upon incubation with HEp-2 cells. We further investigated the effects of Sat on cell lines more relevant to the urinary tract, namely, those derived from bladder and kidney epithelium. Sat elicited elongation of cells and apparent loosening of cellular junctions upon incubation with Vero kidney cells. Additionally, incubation with Sat triggered significant vacuolation within the cytoplasm of both human bladder (CRL-1749) and kidney (CRL-1573) cell lines. This activity has been associated with only a few other known toxins. Following transurethral infection of CBA mice with a sat mutant, no reduction of CFU in urine, bladder, or kidney tissue was seen compared to that in mice infected with wild-type E. coli CFT073. However, significant histological changes were observed within the kidneys of mice infected with wild-type E. coli CFT073, including dissolution of the glomerular membrane and vacuolation of proximal tubule cells. Such damage was not observed in kidney sections of mice infected with a Sat-deficient mutant. These results indicate that Sat, a vacuolating cytotoxin expressed by uropathogenic E. coli CFT073, elicits defined damage to kidney epithelium during upper urinary tract infection and thus contributes to pathogenesis of urinary tract infection.

  14. Interactions between Salmonella typhimurium and Acanthamoeba polyphaga, and observation of a new mode of intracellular growth within contractile vacuoles.

    Science.gov (United States)

    Gaze, W H; Burroughs, N; Gallagher, M P; Wellington, E M H

    2003-10-01

    Acanthamoeba polyphaga feeding on Salmonella typhimurium in a simple model biofilm were observed by light microscopy and a detailed record of interactions kept by digital image capture and image analysis. A strain of S. typhimurium SL1344 carrying a fis: gfp reporter construct (pPDT105) was used to assess intracellular growth in A. polyphaga on non-nutrient agar (NNA) plates. Invasion of the contractile vacuole (CV) was observed at a frequency of 1:100-1000 acanthamoebae at 35 degrees C. The salmonellae contained in CVs illustrated significant up-regulation of fis relative to extracellular bacteria, indicating that they were in the early stages of logarithmic growth, and reached numbers of 100-200 cells per vacuole after 4 days. This is the first report of this mode of intracellular growth. Up-regulation of fis was also observed in a proportion of S. typhimurium cells contained within food vacuoles. Filamentation of S. typhimurium and E. coli cells was frequently observed in coculture with A. polyphaga on NNA plates, with bacterial cells reaching lengths of up to 500 microm after 10 days' incubation at 35 degrees C. A. polyphaga was also seen to mediate bacterial translocation over the agar surface; egested salmonellae subsequently formed microcolonies along amoebal tracks. This illustrated intracellular survival of a fraction of the S. typhimurium population. These phenomena suggest that protozoa such as A. polyhaga may play an important role in the ecology of S. typhimurium in soil and aquatic environments.

  15. Electron tomography characterization of hemoglobin uptake in Plasmodium chabaudi reveals a stage-dependent mechanism for food vacuole morphogenesis.

    Science.gov (United States)

    Wendt, Camila; Rachid, Rachel; de Souza, Wanderley; Miranda, Kildare

    2016-05-01

    In the course of their intraerythrocytic development, malaria parasites incorporate and degrade massive amounts of the host cell cytoplasm. This mechanism is essential for parasite development and represents a physiological step used as target for many antimalarial drugs; nevertheless, the fine mechanisms underlying these processes in Plasmodium species are still under discussion. Here, we studied the events of hemoglobin uptake and hemozoin nucleation in the different stages of the intraerythrocytic cycle of the murine malaria parasite Plasmodium chabaudi using transmission electron tomography of cryofixed and freeze-substituted cells. The results showed that hemoglobin uptake in P. chabaudi starts at the early ring stage and is present in all developmental stages, including the schizont stage. Hemozoin nucleation occurs near the membrane of small food vacuoles. At the trophozoite stage, food vacuoles are found closely localized to cytostomal tubes and mitochondria, whereas in the schizont stage, we observed a large food vacuole located in the central portion of the parasite. Taken together, these results provide new insights into the mechanisms of hemoglobin uptake and degradation in rodent malaria parasites.

  16. Glucose alleviates cadmium toxicity by increasing cadmium fixation in root cell wall and sequestration into vacuole in Arabidopsis

    Institute of Scientific and Technical Information of China (English)

    Yuan-Zhi Shi; Xiao-Fang Zhu; Jiang-Xue Wan; Gui-Xin Li; Shao-Jian Zheng

    2015-01-01

    Glucose (Glu) is involved in not only plant physiological and developmental events but also plant responses to abiotic stresses. Here, we found that the exogenous Glu improved root and shoot growth, reduced shoot cadmium (Cd) concentration, and rescued Cd-induced chlorosis in Arabidopsis thaliana (Columbia ecotype, Col-0) under Cd stressed conditions. Glucose increased Cd retained in the roots, thus reducing its translocation from root to shoot significantly. The most Cd retained in the roots was found in the hemicellulose 1. Glucose combined with Cd (Glu þ Cd) treatment did not affect the content of pectin and its binding capacity of Cd while it increased the content of hemicelluloses 1 and the amount of Cd retained in it significantly. Furthermore, Leadmium Green staining indicated that more Cd was compartmented into vacuoles in Glu þ Cd treatment compared with Cd treatment alone, which was in accordance with the significant upregulation of the expression of tonoplast-localized metal transporter genes, suggesting that com-partmentation of Cd into vacuoles also contributes to the Glu-alleviated Cd toxicity. Taken together, we demonstrated that Glu-alleviated Cd toxicity is mediated through increas-ing Cd fixation in the root cell wall and sequestration into the vacuoles.

  17. Mutations in the Plasmodium falciparum chloroquine resistance transporter, PfCRT, enlarge the parasite's food vacuole and alter drug sensitivities.

    Science.gov (United States)

    Pulcini, Serena; Staines, Henry M; Lee, Andrew H; Shafik, Sarah H; Bouyer, Guillaume; Moore, Catherine M; Daley, Daniel A; Hoke, Matthew J; Altenhofen, Lindsey M; Painter, Heather J; Mu, Jianbing; Ferguson, David J P; Llinás, Manuel; Martin, Rowena E; Fidock, David A; Cooper, Roland A; Krishna, Sanjeev

    2015-01-01

    Mutations in the Plasmodium falciparum chloroquine resistance transporter, PfCRT, are the major determinant of chloroquine resistance in this lethal human malaria parasite. Here, we describe P. falciparum lines subjected to selection by amantadine or blasticidin that carry PfCRT mutations (C101F or L272F), causing the development of enlarged food vacuoles. These parasites also have increased sensitivity to chloroquine and some other quinoline antimalarials, but exhibit no or minimal change in sensitivity to artemisinins, when compared with parental strains. A transgenic parasite line expressing the L272F variant of PfCRT confirmed this increased chloroquine sensitivity and enlarged food vacuole phenotype. Furthermore, the introduction of the C101F or L272F mutation into a chloroquine-resistant variant of PfCRT reduced the ability of this protein to transport chloroquine by approximately 93 and 82%, respectively, when expressed in Xenopus oocytes. These data provide, at least in part, a mechanistic explanation for the increased sensitivity of the mutant parasite lines to chloroquine. Taken together, these findings provide new insights into PfCRT function and PfCRT-mediated drug resistance, as well as the food vacuole, which is an important target of many antimalarial drugs.

  18. Enzymatic Characterization of Recombinant Food Vacuole Plasmepsin 4 from the Rodent Malaria Parasite Plasmodium berghei.

    Directory of Open Access Journals (Sweden)

    Peng Liu

    Full Text Available The rodent malaria parasite Plasmodium berghei is a practical model organism for experimental studies of human malaria. Plasmepsins are a class of aspartic proteinase isoforms that exert multiple pathological effects in malaria parasites. Plasmepsins residing in the food vacuole (FV of the parasite hydrolyze hemoglobin in red blood cells. In this study, we cloned PbPM4, the FV plasmepsin gene of P. berghei that encoded an N-terminally truncated pro-segment and the mature enzyme from genomic DNA. We over-expressed this PbPM4 zymogen as inclusion bodies (IB in Escherichia coli, and purified the protein following in vitro IB refolding. Auto-maturation of the PbPM4 zymogen to mature enzyme was carried out at pH 4.5, 5.0, and 5.5. Interestingly, we found that the PbPM4 zymogen exhibited catalytic activity regardless of the presence of the pro-segment. We determined the optimal catalytic conditions for PbPM4 and studied enzyme kinetics on substrates and inhibitors of aspartic proteinases. Using combinatorial chemistry-based peptide libraries, we studied the active site preferences of PbPM4 at subsites S1, S2, S3, S1', S2' and S3'. Based on these results, we designed and synthesized a selective peptidomimetic compound and tested its inhibition of PbPM4, seven FV plasmepsins from human malaria parasites, and human cathepsin D (hcatD. We showed that this compound exhibited a >10-fold selectivity to PbPM4 and human malaria parasite plasmepsin 4 orthologs versus hcatD. Data from this study furthesr our understanding of enzymatic characteristics of the plasmepsin family and provides leads for anti-malarial drug design.

  19. Sudden collapse of vacuoles in Saintpaulia sp. palisade cells induced by a rapid temperature decrease.

    Directory of Open Access Journals (Sweden)

    Noriaki Kadohama

    Full Text Available It is well known that saintpaulia leaf is damaged by the rapid temperature decrease when cold water is irrigated onto the leaf surface. We investigated this temperature sensitivity and the mechanisms of leaf damage in saintpaulia (Saintpaulia sp. cv. 'Iceberg' and other Gesneriaceae plants. Saintpaulia leaves were damaged and discolored when subjected to a rapid decrease in temperature, but not when the temperature was decreased gradually. Sensitivity to rapid temperature decrease increased within 10 to 20 min during pre-incubation at higher temperature. Injury was restricted to the palisade mesophyll cells, where there was an obvious change in the color of the chloroplasts. During a rapid temperature decrease, chlorophyll fluorescence monitored by a pulse amplitude modulated fluorometer diminished and did not recover even after rewarming to the initial temperature. Isolated chloroplasts were not directly affected by the rapid temperature decrease. Intracellular pH was monitored with a pH-dependent fluorescent dye. In palisade mesophyll cells damaged by rapid temperature decrease, the cytosolic pH decreased and the vacuolar membrane collapsed soon after a temperature decrease. In isolated chloroplasts, chlorophyll fluorescence declined when the pH of the medium was lowered. These results suggest that a rapid temperature decrease directly or indirectly affects the vacuolar membrane, resulting in a pH change in the cytosol that subsequently affects the chloroplasts in palisade mesophyll cells. We further confirmed that the same physiological damage occurs in other Gesneriaceae plants. These results strongly suggested that the vacuoles of palisade mesophyll cells collapsed during the initial phase of leaf injury.

  20. Sudden collapse of vacuoles in Saintpaulia sp. palisade cells induced by a rapid temperature decrease.

    Science.gov (United States)

    Kadohama, Noriaki; Goh, Tatsuaki; Ohnishi, Miwa; Fukaki, Hidehiro; Mimura, Tetsuro; Suzuki, Yoshihiro

    2013-01-01

    It is well known that saintpaulia leaf is damaged by the rapid temperature decrease when cold water is irrigated onto the leaf surface. We investigated this temperature sensitivity and the mechanisms of leaf damage in saintpaulia (Saintpaulia sp. cv. 'Iceberg') and other Gesneriaceae plants. Saintpaulia leaves were damaged and discolored when subjected to a rapid decrease in temperature, but not when the temperature was decreased gradually. Sensitivity to rapid temperature decrease increased within 10 to 20 min during pre-incubation at higher temperature. Injury was restricted to the palisade mesophyll cells, where there was an obvious change in the color of the chloroplasts. During a rapid temperature decrease, chlorophyll fluorescence monitored by a pulse amplitude modulated fluorometer diminished and did not recover even after rewarming to the initial temperature. Isolated chloroplasts were not directly affected by the rapid temperature decrease. Intracellular pH was monitored with a pH-dependent fluorescent dye. In palisade mesophyll cells damaged by rapid temperature decrease, the cytosolic pH decreased and the vacuolar membrane collapsed soon after a temperature decrease. In isolated chloroplasts, chlorophyll fluorescence declined when the pH of the medium was lowered. These results suggest that a rapid temperature decrease directly or indirectly affects the vacuolar membrane, resulting in a pH change in the cytosol that subsequently affects the chloroplasts in palisade mesophyll cells. We further confirmed that the same physiological damage occurs in other Gesneriaceae plants. These results strongly suggested that the vacuoles of palisade mesophyll cells collapsed during the initial phase of leaf injury.

  1. Light-induced morphological alteration in anthocyanin-accumulating vacuoles of maize cells

    Directory of Open Access Journals (Sweden)

    Grotewold Erich

    2005-05-01

    Full Text Available Abstract Background Plant pigmentation is affected by a variety of factors. Light, an important plant developmental signal, influences the accumulation of anthocyanins primarily through the activation of the transcription factors that regulate the flavonoid biosynthetic pathway. In this study, we utilized maize Black Mexican Sweet (BMS cells expressing the R and C1 regulators of anthocyanin biosynthesis from a light-insensitive promoter as a means to investigate the existence of additional levels of control of pigmentation by light. Results BMS cells expressing the R and C1 regulators from the CaMV 35S constitutive promoter accumulate anthocyanins when grown in complete darkness, suggesting that the transcription factors R and C1 are sufficient for the transcription of the genes corresponding to the structural enzymes of the pathway, with no requirement for additional light-induced regulators. Interestingly, light induces a "darkening" in the color of the purple anthocyanin pigmentation of transgenic BMS cells expressing R and C1. This change in the pigment hue is not associated with a variation in the levels or types of anthocyanins present, or with an alteration of the transcript levels of several flavonoid biosynthetic genes. However, cytological observations show that light drives unexpected changes in the morphology and distribution of the anthocyanins-containing vacuolar compartments. Conclusion By uncoupling the effect of light on anthocyanin accumulation, we have found light to induce the fusion of anthocyanin-containing vacuoles, the coalescence of anthocyanic vacuolar inclusion (AVI-like structures contained, and the spread of anthocyanins from the inclusions into the vacuolar sap. Similar light-induced alterations in vacuolar morphology are also evident in the epidermal cells of maize floral whorls accumulating anthocyanins. Our findings suggest a novel mechanism for the action of light on the vacuolar storage of anthocyanin.

  2. Effect of Trimethyltin Chloride on Slow Vacuolar (SV) Channels in Vacuoles from Red Beet (Beta vulgaris L.) Taproots.

    Science.gov (United States)

    Trela, Zenon; Burdach, Zbigniew; Siemieniuk, Agnieszka; Przestalski, Stanisław; Karcz, Waldemar

    2015-01-01

    In the present study, patch-clamp techniques have been used to investigate the effect of trimethyltin chloride (Met3SnCl) on the slow vacuolar (SV) channels in vacuoles from red beet (Beta vulgaris L.) taproots. Activity of SV channels has been measured in whole-vacuole and cytosolic side-out patch configurations. It was found that addition of trimethyltin chloride to the bath solution suppressed, in a concentration-dependent manner, SV currents in red beet vacuoles. The time constant, τ, increased significantly in the presence of the organotin. When single channel activity was analyzed, only little channel activity could be recorded at 100 μM Met3SnCl. Trimethyltin chloride added to the bath medium significantly decreased (by ca. threefold at 100 μM Met3SnCl and at 100 mV voltage, as compared to the control medium) the open probability of single channels. Single channel recordings obtained in the presence and absence of trimethyltin chloride showed that the organotin only slightly (by <10%) decreased the unitary conductance of single channels. It was also found that Met3SnCl significantly diminished the number of SV channel openings, whereas it did not change the opening times of the channels. Taking into account the above and the fact that under the here applied experimental conditions (pH = 7.5) Met3SnCl is a non-dissociated (more lipophilic) compound, we suggest that the suppression of SV currents observed in the presence of the organotin results probably from its hydrophobic properties allowing this compound to translocate near the selectivity filter of the channel.

  3. Pulp tissue vacuolization and necrosis after direct pulp capping with calcium hydroxide and transforming growth factor-β1

    Directory of Open Access Journals (Sweden)

    Sri Kunarti

    2008-03-01

    Full Text Available Mechanical pulp exposure by a rotary cutting instrument or a hand-cutting instrument often happens in deep caries. Application of protective dressing can protect the pulp from additional injury by facilitating healing and repair. Pulp capping has been suggested as one treatment of choice after pulp exposure to maintain pulp vitality. TGF-β1 is growth factor that has important rule in wound healing. The application of Ca(OH2 and exogenous TGF-β1 as direct pulp capping tr4eatment must be experimented in-vivo to see the vacuolization and necrosis in 7, 14, and 21 days after application. This research was done in vivo experiment from orthodontic patients indicated for premolar extraction, between ages 10–15 years. A class V cavity preparation was created in the buccal aspect 1 mm above gingival margin until pulp exposure. Cavity was irrigated slowly with saline solution and dried with a sterile small cotton pellet. Group 1 calcium hydroxide was applied as manufacture procedure. Group 2, the sterile absorbable collagen membrane used, as inert carrier of TGF-β1 was soaked with 5 ml. All groups were covered by a Teflon pledge to separate pulp capping agent from glass ionomer cement restoration. Teeth extracted in 7, 14 and 21 days after treatment. All samples were hystopathologically examined. There were significant difference of TGF-β1 (p < 0.05 in the vacuolization day 14th and 21th compared with 7th. there were not significant difference in necrosis for all variables. Vacuolization and necrosis decreased in the application of TGF-β1.

  4. Analysis of a β-helical region in the p55 domain of Helicobacter pylori vacuolating toxin

    Directory of Open Access Journals (Sweden)

    Algood Holly

    2010-02-01

    Full Text Available Abstract Background Helicobacter pylori is a gram-negative bacterium that colonizes the human stomach and contributes to the development of gastric cancer and peptic ulcer disease. VacA, a toxin secreted by H. pylori, is comprised of two domains, designated p33 and p55. Analysis of the crystal structure of the p55 domain indicated that its structure is predominantly a right-handed parallel β-helix, which is a characteristic of autotransporter passenger domains. Substitution mutations of specific amino acids within the p33 domain abrogate VacA activity, but thus far, it has been difficult to identify small inactivating mutations within the p55 domain. Therefore, we hypothesized that large portions of the p55 domain might be non-essential for vacuolating toxin activity. To test this hypothesis, we introduced eight deletion mutations (each corresponding to a single coil within a β-helical segment spanning VacA amino acids 433-628 into the H. pylori chromosomal vacA gene. Results All eight of the mutant VacA proteins were expressed by the corresponding H. pylori mutant strains and underwent proteolytic processing to yield ~85 kDa passenger domains. Three mutant proteins (VacA Δ484-504, Δ511-536, and Δ517-544 were secreted and induced vacuolation of mammalian cells, which indicated that these β-helical coils were dispensable for vacuolating toxin activity. One mutant protein (VacA Δ433-461 exhibited reduced vacuolating toxin activity compared to wild-type VacA. Other mutant proteins, including those containing deletions near the carboxy-terminal end of the β-helical region (amino acids Val559-Asn628, exhibited marked defects in secretion and increased susceptibility to proteolytic cleavage by trypsin, which suggested that these proteins were misfolded. Conclusions These results indicate that within the β-helical segment of the VacA p55 domain, there are regions of plasticity that tolerate alterations without detrimental effects on protein

  5. Effects of the disaggregation of high-polymerized particles in guard cell vacuoles on osmoregulation of stomatal aperture (stomata opening)

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    Observation under an electron microscope reveals that in closed and open stomata of V. faba, the average volume of particles in guard cell vacuoles (GCV) reduces about 3 orders in magnitude, while the distribution density of the particles increases about 2 orders of magnitude. By using the method of the ratio of fluorescent emissions with laser scanning confocal microscopy, the monitoring to stomata opening shows that during 10 to 30 s before the first distinguishable aperture of stomata, there is a change of pH in GCV about-0.5 units. A quick stomatal opening immediately follows the changes of pH in GCV to reach a steady aperture about 12μm in 100-200 s. This work proposes a model for the osmoregulation in GCV for stomatal opening. The proposed osmoregulation is related to the disaggregation of some polymerized particles inside GCV, which is probably induced by a -(pH in the vacuole. This model describes a process of osmoregulation that avoids the massive energy consuming transportation across cell membranes, which is a foundation of the current chemiosmotic hypothesis. This model is a supplement to the multiple controlling hypothesis for the stomatal movement, which widens research principle ideas for other quick movements in plants.

  6. Virulence-related Mycobacterium avium subsp hominissuis MAV_2928 gene is associated with vacuole remodeling in macrophages

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    Vogt Steven

    2010-04-01

    Full Text Available Abstract Background Mycobacterium avium subsp hominissuis (previously Mycobacterium avium subsp avium is an environmental organism associated with opportunistic infections in humans. Mycobacterium hominissuis infects and replicates within mononuclear phagocytes. Previous study characterized an attenuated mutant in which the PPE gene (MAV_2928 homologous to Rv1787 was inactivated. This mutant, in contrast to the wild-type bacterium, was shown both to have impaired the ability to replicate within macrophages and to have prevented phagosome/lysosome fusion. Results MAV_2928 gene is primarily upregulated upon phagocytosis. The transcriptional profile of macrophages infected with the wild-type bacterium and the mutant were examined using DNA microarray, which showed that the two bacteria interact uniquely with mononuclear phagocytes. Based on the results, it was hypothesized that the phagosome environment and vacuole membrane of the wild-type bacterium might differ from the mutant. Wild-type bacterium phagosomes expressed a number of proteins different from those infected with the mutant. Proteins on the phagosomes were confirmed by fluorescence microscopy and Western blot. The environment in the phagosome of macrophages infected with the mutant differed from the environment of vacuoles with M. hominissuis wild-type in the concentration of zinc, manganese, calcium and potassium. Conclusion The results suggest that the MAV_2928 gene/operon might participate in the establishment of bacterial intracellular environment in macrophages.

  7. Dot/Icm Effector Translocation by Legionella longbeachae Creates a Replicative Vacuole Similar to That of Legionella pneumophila despite Translocation of Distinct Effector Repertoires.

    Science.gov (United States)

    Wood, Rebecca E; Newton, Patrice; Latomanski, Eleanor A; Newton, Hayley J

    2015-10-01

    Legionella organisms are environmental bacteria and accidental human pathogens that can cause severe pneumonia, termed Legionnaires' disease. These bacteria replicate within a pathogen-derived vacuole termed the Legionella-containing vacuole (LCV). Our understanding of the development and dynamics of this vacuole is based on extensive analysis of Legionella pneumophila. Here, we have characterized the Legionella longbeachae replicative vacuole (longbeachae-LCV) and demonstrated that, despite important genomic differences, key features of the replicative LCV are comparable to those of the LCV of L. pneumophila (pneumophila-LCV). We constructed a Dot/Icm-deficient strain by deleting dotB and demonstrated the inability of this mutant to replicate inside THP-1 cells. L. longbeachae does not enter THP-1 cells as efficiently as L. pneumophila, and this is reflected in the observation that translocation of BlaM-RalFLLO (where RalFLLO is the L. longbeachae homologue of RalF) into THP-1 cells by the L. longbeachae Dot/Icm system is less efficient than that by L. pneumophila. This difference is negated in A549 cells where L. longbeachae and L. pneumophila infect with similar entry dynamics. A β-lactamase assay was employed to demonstrate the translocation of a novel family of proteins, the Rab-like effector (Rle) proteins. Immunofluorescence analysis confirmed that these proteins enter the host cell during infection and display distinct subcellular localizations, with RleA and RleC present on the longbeachae-LCV. We observed that the host Rab GTPase, Rab1, and the v-SNARE Sec22b are also recruited to the longbeachae-LCV during the early stages of infection, coinciding with the LCV avoiding endocytic maturation. These studies further our understanding of the L. longbeachae replicative vacuole, highlighting phenotypic similarities to the vacuole of L. pneumophila as well as unique aspects of LCV biology.

  8. Three v-SNAREs and two t-SNAREs, present in a pentameric cis-SNARE complex on isolated vacuoles, are essential for homotypic fusion

    DEFF Research Database (Denmark)

    Ungermann, C; von Mollard, G F; Jensen, Ole Nørregaard

    1999-01-01

    in the same cis multi-SNARE complex. After priming, which disassembles the cis-SNARE complex, antibodies to any of the five SNARE proteins still inhibit the fusion assay until the docking stage is completed, suggesting that each SNARE plays a role in docking. Furthermore, vti1 temperature-sensitive alleles...... cause a synthetic fusion-defective phenotype in our reaction. Our data show that vacuole-vacuole fusion requires a cis-SNARE complex of five SNAREs, the t-SNAREs Vam3p and Vam7p and the v-SNAREs Nyv1p, Vti1p, and Ykt6p....

  9. Liver-specific Aquaporin 11 knockout mice show rapid vacuolization of the rough endoplasmic reticulum in periportal hepatocytes after feeding amino acids

    DEFF Research Database (Denmark)

    Rojek, Aleksandra; Füchtbauer, Ernst-Martin; Füchtbauer, Annette

    2013-01-01

    of the mice (24 h) induced modest dilatation of the rough endoplasmic reticulum (RER) in the periportal hepatocytes. Refeeding with standard mouse chow induced rapid generation of large RER-derived vacuoles in Aqp11 KO mice hepatocytes. Similar effects were observed following oral administration of pure...... protein or larger doses of various amino acids. The fasting/refeeding challenge is associated with increased expression of markers of ER stress Grp78 and GADD153 and decreased glutathione levels, suggesting that ER stress may play role in the development of vacuoles in the AQP11-deficient hepatocytes. NMR...

  10. Complementary DNA display selection of high-affinity peptides binding the vacuolating toxin (VacA) of Helicobacter pylori.

    Science.gov (United States)

    Hayakawa, Yumiko; Matsuno, Mitsuhiro; Tanaka, Makoto; Wada, Akihiro; Kitamura, Koichiro; Takei, Osamu; Sasaki, Ryuzo; Mizukami, Tamio; Hasegawa, Makoto

    2015-09-01

    Artificial peptides designed for molecular recognition of a bacterial toxin have been developed. Vacuolating cytotoxin A protein (VacA) is a major virulence factor of Helicobacter pylori, a gram-negative microaerophilic bacterium inhabiting the upper gastrointestinal tract, particularly the stomach. This study attempted to identify specific peptide sequences with high affinity for VacA using systematic directed evolution in vitro, a cDNA display method. A surface plasmon resonance-based biosensor and fluorescence correlation spectroscopy to examine binding of peptides with VacA identified a peptide (GRVNQRL) with high affinity. Cyclization of the peptide by attaching cysteine residues to both termini improved its binding affinity to VacA, with a dissociation constant (Kd ) of 58 nm. This study describes a new strategy for the development of artificial functional peptides, which are promising materials in biochemical analyses and medical applications.

  11. Inhibitory monoclonal antibody against a (myristylated) small-molecular-weight antigen from Plasmodium falciparum associated with the parasitophorous vacuole membrane.

    Science.gov (United States)

    Kara, U A; Stenzel, D J; Ingram, L T; Bushell, G R; Lopez, J A; Kidson, C

    1988-04-01

    A small-molecular-weight antigen that occurs in asexual blood stages in synchronized cultures of Plasmodium falciparum was detected by a monoclonal antibody which inhibits parasite growth in vitro. This antigen, QF116, showed a molecular weight of 15,000 in parasite strain FCR-3K+ from The Gambia and 19,000 in strain FCQ-27 from Papua New Guinea. The protein did not show significant glycosylation by galactose or glucosamine labeling but was found to be acylated by myristic acid. By using immunogold labeling and electron microscopy, the location of the antigen could be attributed to the parasitophorous vacuole membrane and to inclusions and vesicles residing within the cytoplasm of the erythrocyte host cell.

  12. The nucleocytoplasmic microfilament network in protoplasts from cultured soybean cells is a plastic entity that pervades the cytoplasm except the central vacuole.

    Science.gov (United States)

    Villanueva, Marco A; Schindler, Melvin; Wang, John L

    2005-11-01

    The microfilament network of cultured Glycine max cells (SB-1 line), and protoplasts was visualized with rhodamine-phalloidin under conditions that lysed the protoplast and changed the cell shape. The whole cell had the typical microfilament distribution of a "cage" around the nucleus, from which the large subcortical cables and transvacuolar strands radiated towards the cortex until it reached the cortical microfilament network. Upon cell wall removal, the network conserved its compartmentalization. Thus, the redistribution of the shape where the vacuole becomes a central entity, made the cytoplasm displace peripherally, but the network distribution was conserved. When protoplasts were lysed in a low osmotic medium, the vacuoles were gradually released intact. Under these conditions, the F-actin staining remained within the ghost of the cell, but none was detected in either emerging or almost completely released vacuoles. Most of the released F-actin was found in debris from the cell lysate in the form of microfilaments. When the ghosts were constrained in a coverslip with an air bubble, the shape of the ghost changed accordingly, but the microfilament network distribution remained constant. These results provide further evidence that the vacuole of plant cells does not have detectable associated F-actin. In addition, we demonstrate that the actin microfilament network is a moldable entity that can change its shape but keeps its distribution under constant conditions, in these cultured cells.

  13. Effects of stereochemistry, saturation, and hydrocarbon chain length on the ability of synthetic constrained azacyclic sphingolipids to trigger nutrient transporter down-regulation, vacuolation, and cell death.

    Science.gov (United States)

    Perryman, Michael S; Tessier, Jérémie; Wiher, Timothy; O'Donoghue, Heather; McCracken, Alison N; Kim, Seong M; Nguyen, Dean G; Simitian, Grigor S; Viana, Matheus; Rafelski, Susanne; Edinger, Aimee L; Hanessian, Stephen

    2016-09-15

    Constrained analogs containing a 2-hydroxymethylpyrrolidine core of the natural sphingolipids sphingosine, sphinganine, N,N-dimethylsphingosine and N-acetyl variants of sphingosine and sphinganine (C2-ceramide and dihydro-C2-ceramide) were synthesized and evaluated for their ability to down-regulate nutrient transporter proteins and trigger cytoplasmic vacuolation in mammalian cells. In cancer cells, the disruptions in intracellular trafficking produced by these sphingolipids lead to cancer cell death by starvation. Structure activity studies were conducted by varying the length of the hydrocarbon chain, the degree of unsaturation and the presence or absence of an aryl moiety on the appended chains, and stereochemistry at two stereogenic centers. In general, cytotoxicity was positively correlated with nutrient transporter down-regulation and vacuolation. This study was intended to identify structural and functional features in lead compounds that best contribute to potency, and to develop chemical biology tools that could be used to isolate the different protein targets responsible for nutrient transporter loss and cytoplasmic vacuolation. A molecule that produces maximal vacuolation and transporter loss is expected to have the maximal anti-cancer activity and would be a lead compound.

  14. Acyl-CoA-binding protein, Acb1p, is required for normal vacuole function and ceramide synthesis in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Færgeman, Nils J.; Feddersen, Søren; Christiansen, Janne K;

    2004-01-01

    -sensitive fusion protein attachment protein receptors) Nyv1p, Vam3p and Vti1p, and are unable to fuse in vitro. Mass spectrometric analysis revealed a dramatic reduction in the content of ceramides in whole-cell lipids and in vacuoles isolated from Acb1p-depleted cells. Maturation of yeast aminopeptidase I...

  15. Effector Protein Cig2 Decreases Host Tolerance of Infection by Directing Constitutive Fusion of Autophagosomes with the Coxiella-Containing Vacuole

    Directory of Open Access Journals (Sweden)

    Lara J. Kohler

    2016-07-01

    Full Text Available Coxiella burnetii replicates in an acidified lysosome-derived vacuole. Biogenesis of the Coxiella-containing vacuole (CCV requires bacterial effector proteins delivered into host cells by the Dot/Icm secretion system. Genetic and cell biological analysis revealed that an effector protein called Cig2 promotes constitutive fusion of autophagosomes with the CCV to maintain this compartment in an autolysosomal stage of maturation. This distinguishes the CCV from other pathogen-containing vacuoles that are targeted by the host autophagy pathway, which typically confers host resistance to infection by delivering the pathogen to a toxic lysosomal environment. By maintaining the CCV in an autolysosomal stage of maturation, Cig2 enabled CCV homotypic fusion and enhanced bacterial virulence in the Galleria mellonella (wax moth model of infection by a mechanism that decreases host tolerance. Thus, C. burnetii residence in an autolysosomal organelle alters host tolerance of infection, which indicates that Cig2-dependent manipulation of a lysosome-derived vacuole influences the host response to infection.

  16. Dictyostelium discoideum RabS and Rab2 colocalize with the Golgi and contractile vacuole system and regulate osmoregulation

    Indian Academy of Sciences (India)

    Katherine Maringer; Azure Yarbrough; Sunder Sims-Lucas; Entsar Saheb; Sanaa Jawed; John Bush

    2016-06-01

    Small-molecular-weight GTPase Rab2 has been shown to be a resident of pre-Golgi intermediates and is required for protein transport from the ER to the Golgi complex; however, Rab2 has yet to be characterized in Dictyostelium discoideum. DdRabS is a Dictyostelium Rab that is 80% homologous to DdRab1 which is required for protein transport between the ER and Golgi. Expression of GFP-tagged DdRab2 and DdRabS proteins showed localization to Golgi membranes and to the contractile vacuole system (CV) in Dictyostelium. Microscopic imaging indicates that the DdRab2 and DdRabS proteins localize at, and are essential for, the proper structure of Golgi membranes and the CV system. Dominant negative (DN) forms show fractionation of Golgi membranes, supporting their role in the structure and function of it. DdRab2 and DdRabS proteins, and their dominant negative and constitutively active (CA) forms, affect osmoregulation of the cells, possibly by the influx and discharge of fluids, which suggests a role in the function of the CV system. This is the first evidence of GTPases being localized to both Golgi membranes and the CV system in Dictyostelium.

  17. Functional analysis of Plasmodium falciparum parasitophorous vacuole membrane protein (Pfs16) during gametocytogenesis and gametogenesis by targeted gene disruption.

    Science.gov (United States)

    Kongkasuriyachai, Darin; Fujioka, Hisashi; Kumar, Nirbhay

    2004-02-01

    Gametocytogenesis is a tightly regulated process marked by differentiation through distinct morphological forms and coordinated expression of sexual stage gene products. The earliest known gene product expressed at the onset of Plasmodium falciparum gametocytogenesis is Pfs16 localized on the parasitophorous vacuole membrane (PVM). Targeted gene disruption was undertaken to disrupt expression of Pfs16 and examine its potential role during sexual development. Three independent clones were demonstrated to have the coding sequence of Ps16 gene disrupted by the targeting plasmid by homologous recombination. No full-length transcripts and PVM localized 16 kDa protein were detected. Instead, all three "16ko" clones expressed a protein of 14 kDa recognized by Pfs16 specific antibodies that was mislocalized to an unidentified double membrane compartment in the parasites. Disruption of Pfs16 gene resulted in a significant reduction in gametocyte production, although the small number of gametocytes produced appeared to be normal by molecular and phenotypic evidences. Preliminary observation also suggested impaired ability of male gametocytes to exflagellate in vitro. Pfs16 does not appear to be essential for sexual development, instead may be required for optimal production of sexual parasites. Understanding mechanisms involved in the development of sexual stages of P. falciparum may identify novel targets for drugs and vaccines effective in reducing malaria transmission.

  18. Polyketide synthase (PKS) reduces fusion of Legionella pneumophila-containing vacuoles with lysosomes and contributes to bacterial competitiveness during infection.

    Science.gov (United States)

    Shevchuk, Olga; Pägelow, Dennis; Rasch, Janine; Döhrmann, Simon; Günther, Gabriele; Hoppe, Julia; Ünal, Can Murat; Bronietzki, Marc; Gutierrez, Maximiliano Gabriel; Steinert, Michael

    2014-11-01

    L. pneumophila-containing vacuoles (LCVs) exclude endocytic and lysosomal markers in human macrophages and protozoa. We screened a L. pneumophila mini-Tn10 transposon library for mutants, which fail to inhibit the fusion of LCVs with lysosomes by loading of the lysosomal compartment with colloidal iron dextran, mechanical lysis of infected host cells, and magnetic isolation of LCVs that have fused with lysosomes. In silico analysis of the mutated genes, D. discoideum plaque assays and infection assays in protozoa and U937 macrophage-like cells identified well established as well as novel putative L. pneumophila virulence factors. Promising candidates were further analyzed for their co-localization with lysosomes in host cells using fluorescence microscopy. This approach corroborated that the O-methyltransferase, PilY1, TPR-containing protein and polyketide synthase (PKS) of L. pneumophila interfere with lysosomal degradation. Competitive infections in protozoa and macrophages revealed that the identified PKS contributes to the biological fitness of pneumophila strains and may explain their prevalence in the epidemiology of Legionnaires' disease.

  19. The infectious intracellular lifestyle of Salmonella enterica relies on the adaptation to nutritional conditions within the Salmonella-containing vacuole.

    Science.gov (United States)

    Diacovich, Lautaro; Lorenzi, Lucía; Tomassetti, Mauro; Méresse, Stéphane; Gramajo, Hugo

    2016-12-09

    Salmonella enterica serovar Typhimurium (S. Typhimurium) is a Gram-negative pathogen that causes various host-specific diseases. During their life cycle, Salmonellae survive frequent exposures to a variety of environmental stresses, e.g. carbon-source starvation. The virulence of this pathogen relies on its ability to establish a replicative niche, named Salmonella-containing vacuole, inside host cells. However, the microenvironment of the SCV and the bacterial metabolic pathways required during infection are largely undefined. In this work we developed different biological probes whose expression is modulated by the environment and the physiological state of the bacterium. We constructed transcriptional reporters by fusing promoter regions to the gfpmut3a gene to monitor the expression profile of genes involved in glucose utilization and lipid catabolism. The induction of these probes by a specific metabolic change was first tested in vitro, and then during different conditions of infection in macrophages. We were able to determine that Entner-Doudoroff is the main metabolic pathway utilized by Salmonella during infection in mouse macrophages. Furthermore, we found sub-populations of bacteria expressing genes involved in pathways for the utilization of different sources of carbon. These populations are modified in presence of different metabolizable substrates, suggesting the coexistence of Salmonella with diverse metabolic states during the infection.

  20. Live Cell Imaging During Germination Reveals Dynamic Tubular Structures Derived from Protein Storage Vacuoles of Barley Aleurone Cells

    Directory of Open Access Journals (Sweden)

    Verena Ibl

    2014-09-01

    Full Text Available The germination of cereal seeds is a rapid developmental process in which the endomembrane system undergoes a series of dynamic morphological changes to mobilize storage compounds. The changing ultrastructure of protein storage vacuoles (PSVs in the cells of the aleurone layer has been investigated in the past, but generally this involved inferences drawn from static pictures representing different developmental stages. We used live cell imaging in transgenic barley plants expressing a TIP3-GFP fusion protein as a fluorescent PSV marker to follow in real time the spatially and temporally regulated remodeling and reshaping of PSVs during germination. During late-stage germination, we observed thin, tubular structures extending from PSVs in an actin-dependent manner. No extensions were detected following the disruption of actin microfilaments, while microtubules did not appear to be involved in the process. The previously-undetected tubular PSV structures were characterized by complex movements, fusion events and a dynamic morphology. Their function during germination remains unknown, but might be related to the transport of solutes and metabolites.

  1. TcPho91 is a contractile vacuole phosphate sodium symporter that regulates phosphate and polyphosphate metabolism in Trypanosoma cruzi.

    Science.gov (United States)

    Jimenez, Veronica; Docampo, Roberto

    2015-09-01

    We have identified a phosphate transporter (TcPho91) localized to the bladder of the contractile vacuole complex (CVC) of Trypanosoma cruzi, the etiologic agent of Chagas disease. TcPho91 has 12 transmembrane domains, an N-terminal regulatory SPX (named after SYG1, Pho81 and XPR1) domain and an anion permease domain. Functional expression in Xenopus laevis oocytes followed by two-electrode voltage clamp showed that TcPho91 is a low-affinity transporter with a Km for Pi in the millimolar range, and sodium-dependency. Epimastigotes overexpressing TcPho91-green fluorescent protein have significantly higher levels of pyrophosphate (PPi ) and short-chain polyphosphate (polyP), suggesting accumulation of Pi in these cells. Moreover, when overexpressing parasites were maintained in a medium with low Pi , they grew at higher rates than control parasites. Only one allele of TcPho91 in the CL strain encodes for the complete open reading frame, while the other one is truncated encoding for only the N-terminal domain. Taking advantage of this characteristic, knockdown experiments were performed resulting in cells with reduced growth rate as well as a reduction in PPi and short-chain polyP levels. Our results indicate that TcPho91 is a phosphate sodium symporter involved in Pi homeostasis in T. cruzi.

  2. Toxoplasma exports dense granule proteins beyond the vacuole to the host cell nucleus and rewires the host genome expression.

    Science.gov (United States)

    Bougdour, Alexandre; Tardieux, Isabelle; Hakimi, Mohamed-Ali

    2014-03-01

    Toxoplasma gondii is the most widespread apicomplexan parasite and occupies a large spectrum of niches by infecting virtually any warm-blooded animals. As an obligate intracellular parasite, Toxoplasma has evolved a repertoire of strategies to fine-tune the cellular environment in an optimal way to promote growth and persistence in host tissues hence increasing the chance to be transmitted to new hosts. Short and long-term intracellular survival is associated with Toxoplasma ability to both evade the host deleterious immune defences and to stimulate a beneficial immune balance by governing host cell gene expression. It is only recently that parasite proteins responsible for driving these transcriptional changes have been identified. While proteins contained in the apical secretory Rhoptry organelle have already been identified as bona fide secreted effectors that divert host signalling pathways, recent findings revealed that dense granule proteins should be added to the growing list of effectors as they reach the host cell cytoplasm and nucleus and target various host cell pathways in the course of cell infection. Herein, we emphasize on a novel subfamily of dense granule residentproteins, exemplified with the GRA16 and GRA24 members we recently discovered as both are exported beyond the vacuole-containing parasites and reach the host cell nucleus to reshape the host genome expression.

  3. Proteolytic Activation of the Essential Parasitophorous Vacuole Cysteine Protease SERA6 Accompanies Malaria Parasite Egress from Its Host Erythrocyte*

    Science.gov (United States)

    Ruecker, Andrea; Shea, Michael; Hackett, Fiona; Suarez, Catherine; Hirst, Elizabeth M. A.; Milutinovic, Katarina; Withers-Martinez, Chrislaine; Blackman, Michael J.

    2012-01-01

    The malaria parasite replicates within an intraerythrocytic parasitophorous vacuole (PV). The PV and host cell membranes eventually rupture, releasing merozoites in a process called egress. Certain inhibitors of serine and cysteine proteases block egress, indicating a crucial role for proteases. The Plasmodium falciparum genome encodes nine serine-repeat antigens (SERAs), each of which contains a central domain homologous to the papain-like (clan CA, family C1) protease family. SERA5 and SERA6 are indispensable in blood-stage parasites, but the function of neither is known. Here we show that SERA6 localizes to the PV where it is precisely cleaved just prior to egress by an essential serine protease called PfSUB1. Mutations that replace the predicted catalytic Cys of SERA6, or that block SERA6 processing by PfSUB1, could not be stably introduced into the parasite genomic sera6 locus, indicating that SERA6 is an essential enzyme and that processing is important for its function. We demonstrate that cleavage of SERA6 by PfSUB1 converts it to an active cysteine protease. Our observations reveal a proteolytic activation step in the malarial PV that may be required for release of the parasite from its host erythrocyte. PMID:22984267

  4. Live Cell Imaging During Germination Reveals Dynamic Tubular Structures Derived from Protein Storage Vacuoles of Barley Aleurone Cells.

    Science.gov (United States)

    Ibl, Verena; Stoger, Eva

    2014-01-01

    The germination of cereal seeds is a rapid developmental process in which the endomembrane system undergoes a series of dynamic morphological changes to mobilize storage compounds. The changing ultrastructure of protein storage vacuoles (PSVs) in the cells of the aleurone layer has been investigated in the past, but generally this involved inferences drawn from static pictures representing different developmental stages. We used live cell imaging in transgenic barley plants expressing a TIP3-GFP fusion protein as a fluorescent PSV marker to follow in real time the spatially and temporally regulated remodeling and reshaping of PSVs during germination. During late-stage germination, we observed thin, tubular structures extending from PSVs in an actin-dependent manner. No extensions were detected following the disruption of actin microfilaments, while microtubules did not appear to be involved in the process. The previously-undetected tubular PSV structures were characterized by complex movements, fusion events and a dynamic morphology. Their function during germination remains unknown, but might be related to the transport of solutes and metabolites.

  5. Targeting host syntaxin-5 preferentially blocks Leishmania parasitophorous vacuole development in infected cells and limits experimental Leishmania infections.

    Science.gov (United States)

    Canton, Johnathan; Kima, Peter E

    2012-10-01

    Our previous observations established a role for syntaxin-5 in the development of Leishmania parasitophorous vacuoles (LPVs). In this study, we took advantage of the recent identification of Retro-2, a small organic molecule that can cause the redistribution of syntaxin-5; we show herein that Retro-2 blocks LPV development within 2 hours of adding it to cells infected with Leishmania amazonensis. In infected cells incubated for 48 hours with Retro-2, LPV development was significantly limited; furthermore, infected cells harbored four to five times fewer parasites than infected cells incubated in vehicle alone. In vivo studies revealed that Retro-2 curbed experimental L. amazonensis infections in a dose-dependent manner. Retro-2 did not have any appreciable effect on the host cell physiological characteristics; furthermore, it had no apparent toxicity in experimental animals. An unexpected, but welcome, finding was that Retro-2 inhibited the replication of Leishmania parasites in axenic cultures. This study is significant because it identifies an endoplasmic reticulum/Golgi SNARE as a potential target for the control of Leishmania infections; moreover, it suggests that small organic molecules can be identified that can selectively disrupt the vesicle fusion machinery that promotes the development of pathogen-containing compartments without exerting toxic effects on the host.

  6. Quantitative trait locus mapping of genes associated with vacuolation in the adrenal X-zone of the DDD/Sgn inbred mouse

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    Suto Jun-ichi

    2012-11-01

    Full Text Available Abstract Background Adrenal gland of mice contains a transient zone between the adrenal cortex and the adrenal medulla: the X-zone. There are clear strain differences in terms of X-zone morphology. Nulliparous females of the inbred mouse DDD strain develop adrenal X-zones containing exclusively vacuolated cells, whereas females of the inbred mouse B6 strain develop X-zones containing only non-vacuolated cells. The X-zone vacuolation is a physiologic process associated with the X-zone degeneration and is tightly regulated by genetic factors. Identification of the genetic factors controlling such strain differences should help analyze the X-zone function. In this study, a quantitative trait locus (QTL analysis for the extent of X-zone vacuolation was performed for two types of F2 female mice: F2Ay mice (F2 mice with the Ay allele and F2 non-Ay mice (F2 mice without the Ay allele. These were produced by crossing B6 females and DDD.Cg-Ay males. DDD.Cg-Ay is a congenic mouse strain for the Ay allele at the agouti locus and is used for this study because a close association between the X-zone morphology and the agouti locus genotype has been suggested. The Ay allele is dominant and homozygous lethal; therefore, living Ay mice are invariably heterozygotes. Results Single QTL scans identified significant QTLs on chromosomes 1, 2, 6, and X for F2 non-Ay mice, and on chromosomes 2, 6, and 12 for F2Ay mice. The QTL on chromosome 2 was considered to be because of the agouti locus, which has been suggested to be associated with X-zone vacuolation. A significant QTL that interacted with the agouti locus was identified on chromosome 8. Conclusions The extent of X-zone vacuolation in DDD females was controlled by multiple genes with complex interactions. The murine X-zone is considered analogous structure to the human fetal zone. Therefore, the results of this study will aid in understanding function of not only of the X-zone but also of the human fetal zone

  7. Transport of proteins to the plant vacuole is not by bulk flow through the secretory system, and requires positive sorting information.

    Science.gov (United States)

    Dorel, C; Voelker, T A; Herman, E M; Chrispeels, M J

    1989-02-01

    Plant cells, like other eukaryotic cells, use the secretory pathway to target proteins to the vacuolar/lysosomal compartment and to the extracellular space. We wished to determine whether the presence of a hydrophobic signal peptide would result in the transport of a reporter protein to vacuoles by bulk flow; to investigate this question, we expressed a chimeric gene in transgenic tobacco. The chimeric gene, Phalb, used for this study consists of the 1,188-bp 5' upstream sequence and the hydrophobic signal sequence of a vacuolar seed protein phytohemagglutinin, and the coding sequence of a cytosolic seed albumin (PA2). The chimeric protein PHALB cross-reacted with antibodies to PA2 and was found in the seeds of the transgenic plants (approximately 0.7% of total protein), but not in the leaves, roots, or flowers. Immunoblot analyses of seed extracts revealed four glycosylated polypeptides ranging in molecular weight from 29,000 to 32,000. The four polypeptides are glycoforms of a single polypeptide of Mr 27,000, and the heterogeneity is due to the presence of high mannose and endoglycosidase H-resistant glycans. The PHALB products reacted with an antiserum specific for complex plant glycans indicating that the glycans had been modified in the Golgi apparatus. Subcellular fractionation of glycerol extracts of mature seeds showed that only small amounts of PHALB accumulated in the protein storage vacuoles of the tobacco seeds. In homogenates made in an isotonic medium, very little PHALB was associated with the organelle fraction containing the endoplasmic reticulum and Golgi apparatus; most of it was in the soluble fraction. We conclude that PHALB passed through the Golgi apparatus, but did not arrive in the vacuoles. Transport to vacuoles is not by a bulk-flow mechanism, once proteins have entered the secretory system, and requires information beyond that provided by a hydrophobic signal peptide.

  8. Abscisic acid prevents the coalescence of protein storage vacuoles by upregulating expression of a tonoplast intrinsic protein gene in barley aleurone.

    Science.gov (United States)

    Lee, Sung-eun; Yim, Hui-kyung; Lim, Mi-na; Yoon, In sun; Kim, Jeong hoe; Hwang, Yong-sic

    2015-03-01

    Tonoplast intrinsic proteins (TIPs) are integral membrane proteins that are known to function in plants as aquaporins. Here, we propose another role for TIPs during the fusion of protein storage vacuoles (PSVs) in aleurone cells, a process that is promoted by gibberellic acid (GA) and prevented by abscisic acid (ABA). Studies of the expression of barley (Hordeum vulgare) TIP genes (HvTIP) showed that GA specifically decreased the abundance of HvTIP1;2 and HvTIP3;1 transcripts, while ABA strongly increased expression of HvTIP3;1. Increased or decreased expression of HvTIP3;1 interfered with the hormonal effects on vacuolation in aleurone protoplasts. HvTIP3;1 gain-of-function experiments delayed GA-induced vacuolation, whereas HvTIP3;1 loss-of-function experiments promoted vacuolation in ABA-treated aleurone cells. These results indicate that TIP plays a key role in preventing the coalescence of small PSVs in aleurone cells. Hormonal regulation of the HvTIP3;1 promoter is similar to the regulation of the endogenous gene, indicating that induction of the transcription of HvTIP3;1 by ABA is a critical factor in the prevention of PSV coalescence in response to ABA. Promoter analysis using deletions and site-directed mutagenesis of sequences identified three cis-acting elements that are responsible for ABA responsiveness in the HvTIP3;1 promoter. Promoter analysis also showed that ABA responsiveness of the HvTIP3;1 promoter is likely to occur via a unique regulatory system distinct from that involving the ABA-response promoter complexes.

  9. Selection of normal spermatozoa with a vacuole-free head (x6300) improves selection of spermatozoa with intact DNA in patients with high sperm DNA fragmentation rates.

    Science.gov (United States)

    Hammoud, I; Boitrelle, F; Ferfouri, F; Vialard, F; Bergere, M; Wainer, B; Bailly, M; Albert, M; Selva, J

    2013-06-01

    Intracytoplasmic morphologically selected sperm injection (IMSI, 6300× magnification with Nomarski contrast) of a normal spermatozoon with a vacuole-free head could improve the embryo's ability to grow to the blastocyst stage and then implant. However, the most relevant indications for IMSI remain to be determined. To evaluate the potential value of IMSI for patients with a high degree of sperm DNA fragmentation (n = 8), different types of spermatozoa were analysed in terms of DNA fragmentation. Motile normal spermatozoa with a vacuole-free head selected at 6300× magnification had a significantly lower mean DNA fragmentation rate (4.1 ± 1.1%, n = 191) than all other types of spermatozoa: non-selected spermatozoa (n = 8000; 26.1 ± 1.5% versus 4.1 ± 1.1%; P sperm DNA fragmentation rates, selection of normal spermatozoa with a vacuole-free head (6300×) yields the greatest likelihood of obtaining spermatozoa with non-fragmented DNA.

  10. Patch clamp studies on root cell vacuoles of a salt-tolerant and a salt-sensitive plantago species : regulation of channel activity by salt stress.

    Science.gov (United States)

    Maathuis, F J; Prins, H B

    1990-01-01

    Plantago media L. and Plantago maritima L. differ in their strategy toward salt stress, a major difference being the uptake and distribution of ions. Patch clamp techniques were applied to root cell vacuoles to study the tonoplast channel characteristics. In both species the major channel found was a 60 to 70 picosiemens channel with a low ion selectivity. The conductance of this channel for Na(+) was the same as for K(+), P(K) (+)/P(Na) (+) = 1, whereas the cation/anion selectivity (P(K) (+)/P(c1) (-)) was about 5. Gating characteristics were voltage and calcium dependent. An additional smaller channel of 25 picosiemens was present in P. maritima. In the whole vacuole configuration, the summation of the single channel currents resulted in slowly activated inward currents (t((1/2)) = 1.2 second). Inwardly directed, ATP-dependent currents could be measured against a DeltapH gradient of 1.5 units over the tonoplast. This observation strongly indicated the physiological intactness of the used vacuoles. The open probability of the tonoplast channels dramatically decreased when plants were grown on NaCl, although single channel conductance and selectivity were not altered.

  11. Human lung epithelial cells contain Mycobacterium tuberculosis in a late endosomal vacuole and are efficiently recognized by CD8⁺ T cells.

    Directory of Open Access Journals (Sweden)

    Melanie J Harriff

    Full Text Available Mycobacterium tuberculosis (Mtb is transmitted via inhalation of aerosolized particles. While alveolar macrophages are thought to play a central role in the acquisition and control of this infection, Mtb also has ample opportunity to interact with the airway epithelium. In this regard, we have recently shown that the upper airways are enriched with a population of non-classical, MR1-restricted, Mtb-reactive CD8⁺ T cells (MAIT cells. Additionally, we have demonstrated that Mtb-infected epithelial cells lining the upper airways are capable of stimulating IFNγ production by MAIT cells. In this study, we demonstrate that airway epithelial cells efficiently stimulate IFNγ release by MAIT cells as well as HLA-B45 and HLA-E restricted T cell clones. Characterization of the intracellular localization of Mtb in epithelial cells indicates that the vacuole occupied by Mtb in epithelial cells is distinct from DC in that it acquires Rab7 molecules and does not retain markers of early endosomes such as Rab5. The Mtb vacuole is also heterogeneous as there is a varying degree of association with Lamp1 and HLA-I. Although the Mtb vacuole shares markers associated with the late endosome, it does not acidify, and the bacteria are able to replicate within the cell. This work demonstrates that Mtb infected lung epithelial cells are surprisingly efficient at stimulating IFNγ release by CD8⁺ T cells.

  12. New insights into globoids of protein storage vacuoles in wheat aleurone using synchrotron soft X-ray microscopy.

    Science.gov (United States)

    Regvar, Marjana; Eichert, Diane; Kaulich, Burkhard; Gianoncelli, Alessandra; Pongrac, Paula; Vogel-Mikus, Katarina; Kreft, Ivan

    2011-07-01

    Mature developed seeds are physiologically and biochemically committed to store nutrients, principally as starch, protein, oils, and minerals. The composition and distribution of elements inside the aleurone cell layer reflect their biogenesis, structural characteristics, and physiological functions. It is therefore of primary importance to understand the mechanisms underlying metal ion accumulation, distribution, storage, and bioavailability in aleurone subcellular organelles for seed fortification purposes. Synchrotron radiation soft X-ray full-field imaging mode (FFIM) and low-energy X-ray fluorescence (LEXRF) spectromicroscopy were applied to characterize major structural features and the subcellular distribution of physiologically important elements (Zn, Fe, Na, Mg, Al, Si, and P). These direct imaging methods reveal the accumulation patterns between the apoplast and symplast, and highlight the importance of globoids with phytic acid mineral salts and walls as preferential storage structures. C, N, and O chemical topographies are directly linked to the structural backbone of plant substructures. Zn, Fe, Na, Mg, Al, and P were linked to globoid structures within protein storage vacuoles with variable levels of co-localization. Si distribution was atypical, being contained in the aleurone apoplast and symplast, supporting a physiological role for Si in addition to its structural function. These results reveal that the immobilization of metals within the observed endomembrane structures presents a structural and functional barrier and affects bioavailability. The combination of high spatial and chemical X-ray microscopy techniques highlights how in situ analysis can yield new insights into the complexity of the wheat aleurone layer, whose precise biochemical composition, morphology, and structural characteristics are still not unequivocally resolved.

  13. Iron-chelating agent, deferasirox, inhibits neutrophil activation and extracellular trap formation.

    Science.gov (United States)

    Kono, Mari; Saigo, Katsuyasu; Yamamoto, Shiori; Shirai, Kohei; Iwamoto, Shuta; Uematsu, Tomoko; Takahashi, Takayuki; Imoto, Shion; Hashimoto, Makoto; Minami, Yosuke; Wada, Atsushi; Takenokuchi, Mariko; Kawano, Seiji

    2016-10-01

    Iron-chelating agents, which are frequently prescribed to transfusion-dependent patients, have various useful biological effects in addition to chelation. Reactive oxygen species (ROS) produced by neutrophils can cause pulmonary endothelial cell damage, which can lead to acute lung injury (ALI). We previously reported that deferasirox (DFS), an iron-chelating agent, inhibits phorbol myristate acetate (PMA) or formyl-methionyl-leucyl-phenylalanine (fMLP)-induced ROS production in neutrophils, in vitro. Here, we investigate whether DFS inhibits vacuolization in neutrophils and neutrophil extracellular trap (NET) formation. Human neutrophils were incubated with DFS and stimulated with PMA or fMLP. Human neutrophils were separated from heparinized peripheral blood using density gradient centrifugation, and subsequently incubated with DFS. After 10 minutes, neutrophils were stimulated by PMA or fMLP. Vacuole formation was observed by electron microscopy. For observing NET formations using microscopes, immunohistological analyses using citrullinated histone H3 and myeloperoxidase antibodies, and SYTOX Green (an impermeable DNA detection dye) staining, were conducted. NET formation was measured as the quantity of double-stranded DNA (dsDNA), using the AccuBlue Broad Range dsDNA Quantitation Kit. DFS (50 μmol/L) inhibited vacuole formation in the cytoplasm and NET formation. Additionally, 5-100 μmol/L concentration of DFS inhibited the release of dsDNA in a dose-independent manner. We demonstrate that DFS inhibits not only ROS production but also vacuolization and NET formation in neutrophils. These results suggest the possibility of protective effects of DFS against NET-related adverse effects, including ALI and thrombosis.

  14. 青霉素处理检查和分离蓝藻细胞液泡%Determination and Isolation of Cell Vacuoles from Blue-green Algae by Penicillin Method

    Institute of Scientific and Technical Information of China (English)

    郭碧薇; 易平; 刘希玲; 郭厚良

    2003-01-01

    Growing in the liquid medium containing penicillin, the cells of the Cyanobacteria,Anabaena 7120, Nostoc flagelliforme, and Synechocystis 6803 were broken and vacuoles were released. Percentage of broken cells declined and percentage of broken cells increased with the growing days of the algae. The percentage of vacuoles to broken cells were respectively 0.7%, 0.8%, and 13.3% in the three types of algae Anabaena 7120, N.flagelliforme and Synechocystis 6803 which had grown for 3 days.

  15. A fluorescent reporter protein containing AtRMR1 domains is targeted to the storage and central vacuoles in Arabidopsis thaliana and tobacco leaf cells.

    Science.gov (United States)

    Scabone, Camila María; Frigerio, Lorenzo; Petruccelli, Silvana

    2011-10-01

    To develop a new strategy to target recombinant proteins to the vacuolar storage system in transgenic plants, the ability of the transmembrane and cytosolic domains of Arabidopsis receptor homology-transmembrane-RING H2-1 (AtRMR1) was evaluated. A secreted version of RFP (secRFP) and a fusion of it to the transmembrane and cytosolic domains of AtRMR1 (RFP-TMCT) were produced and studied both in transient and stable expression assays. Transient expression in leaves of Nicotiana tabacum showed that secRFP is secreted to the apoplast while its fusion to TMCT of AtRMR1 is sufficient to prevent secretion of the reporter. In tobacco leaves, RFP-TMCT reporter showed an endoplasmic reticulum pattern in early expression stages while in late expression stages, it was found in the vacuolar lumen. For the first time, the role of TM and CT domains of AtRMR1 in stable expression in Arabidopsis thaliana is presented; the fusion of TMCT to secRFP is sufficient to sort RFP to the lumen of the central vacuoles in leaves and roots and to the lumen of PSV in cotyledons of mature embryos. In addition, biochemical studies performed in extract from transgenic plants showed that RFP-TMCT is an integral membrane protein. Full-length RFP-TMCT was also found in the vacuolar lumen, suggesting internalization into destination vacuole. Not colocalization of RFP-TMCT with tonoplast and plasma membrane markers were observed. This membrane vacuolar determinant sorting signal could be used for future application in molecular pharming as an alternative means to sort proteins of interest to vacuoles.

  16. Maternal exposure to hexachlorophene targets intermediate-stage progenitor cells in the hippocampal neurogenesis involving myelin vacuolation of cholinergic and glutamatergic inputs in mice.

    Science.gov (United States)

    Kato, Mizuho; Abe, Hajime; Itahashi, Megu; Kikuchihara, Yoh; Kimura, Masayuki; Mizukami, Sayaka; Yoshida, Toshinori; Shibutani, Makoto

    2016-02-01

    Hexachlorophene (HCP) has been shown to induce myelin vacuolation due to intramyelinic edema of the nerve fibers in animal neural tissue. We investigated the maternal exposure effect of HCP on hippocampal neurogenesis in the offspring of pregnant mice supplemented with 0 (control), 33 or 100 ppm HCP in diet from gestational day 6 to day 21 after delivery. On postnatal day (PND) 21, offspring as examined in males exhibited decreased granule cell lineage populations expressing paired box 6, sex-determining region Y-box 2 and eomesodermin in the hippocampal subgranular zone (SGZ) accompanied by myelin vacuolation involving white matter tracts of the hippocampal fimbria at ≥ 33 ppm. However, SGZ cellular populations expressing brain lipid binding protein and doublecortin were unchanged at any dose. Transcript expression of cholinergic receptor genes, Chrna4 and Chrnb2, and glutamate receptor genes, Grm1 and Grin2d, examined at 100 ppm, decreased in the dentate gyrus. HCP exposure did not alter the number of proliferating or apoptotic cells in the SGZ, or reelin- or calcium-binding protein-expressing γ-aminobutyric acid (GABA)ergic interneurons in the dentate hilus, on PND 21 and PND 77. All neurogenesis-related changes observed in HCP-exposed offspring on PND 21 disappeared on PND 77, suggesting that maternal HCP exposure at ≥ 33 ppm reversibly decreased type 2 intermediate-stage progenitor cells in the hippocampal neurogenesis. Myelin vacuolation might be responsible for changes in neurogenesis possibly by reducing nerve conduction velocity of cholinergic inputs from the septal-hippocampal pathway to granule cell lineages and/or GABAergic interneurons, and of glutamatergic inputs to granule cell lineages.

  17. Anaplasma phagocytophilum APH0032 Is Exposed on the Cytosolic Face of the Pathogen-Occupied Vacuole and Co-opts Host Cell SUMOylation

    Science.gov (United States)

    Oki, Aminat T.; Huang, Bernice; Beyer, Andrea R.; May, Levi J.; Truchan, Hilary K.; Walker, Naomi J.; Galloway, Nathan L.; Borjesson, Dori L.; Carlyon, Jason A.

    2016-01-01

    Anaplasma phagocytophilum, a member of the family Anaplasmataceae and the obligate intracellular bacterium that causes granulocytic anaplasmosis, resides in a host cell-derived vacuole. Bacterial proteins that localize to the A. phagocytophilum-occupied vacuole membrane (AVM) are critical host-pathogen interfaces. Of the few bacterial AVM proteins that have been identified, the domains responsible for AVM localization and the host cell pathways that they co-opt are poorly defined. APH0032 is an effector that is expressed and localizes to the AVM late during the infection cycle. Herein, the APH0032 domain that is essential for associating with host cell membranes was mapped. Immunofluorescent labeling of infected cells that had been differentially permeabilized confirmed that APH0032 is exposed on the AVM's cytosolic face, signifying its potential to interface with host cell processes. SUMOylation is the covalent attachment of a member of the small ubiquitin-like modifier (SUMO) family of proteins to lysines in target substrates. Previous work from our laboratory determined that SUMOylation is important for A. phagocytophilum survival and that SUMOylated proteins decorate the AVM. Algorithmic prediction analyses identified APH0032 as a candidate for SUMOylation. Endogenous APH0032 was precipitated from infected cells using a SUMO affinity matrix, confirming that the effector co-opts SUMOylation during infection. APH0032 pronouncedly colocalized with SUMO1, but not SUMO2/3 moieties on the AVM. Ectopic expression of APH0032 in A. phagocytophilum infected host cells significantly boosted the bacterial load. This study delineates the first domain of any Anaplasmataceae protein that is essential for associating with the pathogen-occupied vacuole membrane, demonstrates the importance of APH0032 to infection, and identifies it as the second A. phagocytophilum effector that co-opts SUMOylation, thus underscoring the relevance of this post-translational modification to

  18. Characterization of Helicobacter pylori VacA-containing vacuoles (VCVs), VacA intracellular trafficking and interference with calcium signalling in T lymphocytes.

    Science.gov (United States)

    Kern, Beate; Jain, Utkarsh; Utsch, Ciara; Otto, Andreas; Busch, Benjamin; Jiménez-Soto, Luisa; Becher, Dörte; Haas, Rainer

    2015-12-01

    The human pathogen Helicobacter pylori colonizes half of the global population. Residing at the stomach epithelium, it contributes to the development of diseases such as gastritis, duodenal and gastric ulcers, and gastric cancer. A major factor is the secreted vacuolating toxin VacA, which forms anion-selective channels in the endosome membrane that cause the compartment to swell, but the composition and purpose of the resulting VacA-containing vacuoles (VCVs) are still unknown. VacA exerts influence on the host immune response in various ways, including inhibition of T-cell activation and proliferation and suppression of the host immune response. In this study, for the first time the composition of VCVs from T cells was comprehensively analysed to investigate VCV function. VCVs were successfully isolated via immunomagnetic separation, and the purified vacuoles were analysed by mass spectrometry. We detected a set of 122 VCV-specific proteins implicated among others in immune response, cell death and cellular signalling processes, all of which VacA is known to influence. One of the individual proteins studied further was stromal interaction molecule (STIM1), a calcium sensor residing in the endoplasmic reticulum (ER) that is important in store-operated calcium entry. Live cell imaging microscopy data demonstrated colocalization of VacA with STIM1 in the ER and indicated that VacA may interfere with the movement of STIM1 towards the plasma membrane-localized calcium release activated calcium channel protein ORAI1 in response to Ca(2+) store depletion. Furthermore, VacA inhibited the increase of cytosolic-free Ca(2+) in the Jurkat E6-1 T-cell line and human CD4(+) T cells. The presence of VacA in the ER and its trafficking to the Golgi apparatus was confirmed in HeLa cells, identifying these two cellular compartments as novel VacA target structures.

  19. Toxoplasma gondii sporozoites form a transient parasitophorous vacuole that is impermeable and contains only a subset of dense-granule proteins.

    OpenAIRE

    Tilley, M; Fichera, M E; Jerome, M E; Roos, D. S.; White, M W

    1997-01-01

    Toxoplasma gondii sporozoites form two parasitophorous vacuoles during development within host cells, the first (PV1) during host cell invasion and the second (PV2) 18 to 24 h postinoculation. PV1 is structurally distinctive due to its large size, yet it lacks a tubulovesicular network (C. A. Speer, M. Tilley, M. Temple, J. A. Blixt, J. P. Dubey, and M. W. White, Mol. Biochem. Parasitol. 75:75-86, 1995). Confirming the finding that sporozoites have a different electron-dense-granule compositi...

  20. Plasmodium berghei Δp52&p36 parasites develop independent of a parasitophorous vacuole membrane in Huh-7 liver cells.

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    Ivo H J Ploemen

    Full Text Available The proteins P52 and P36 are expressed in the sporozoite stage of the murine malaria parasite Plasmodium berghei. Δp52&p36 sporozoites lacking expression of both proteins are severely compromised in their capability to develop into liver stage parasites and abort development soon after invasion; presumably due to the absence of a parasitophorous vacuole membrane (PVM. However, a small proportion of P. berghei Δp52&p36 parasites is capable to fully mature in hepatocytes causing breakthrough blood stage infections. We have studied the maturation of replicating Δp52&p36 parasites in cultured Huh-7 hepatocytes. Approximately 50% of Δp52&p36 parasites developed inside the nucleus of the hepatocyte but did not complete maturation and failed to produce merosomes. In contrast cytosolic Δp52&p36 parasites were able to fully mature and produced infectious merozoites. These Δp52&p36 parasites developed into mature schizonts in the absence of an apparent parasitophorous vacuole membrane as shown by immunofluorescence and electron microscopy. Merozoites derived from these maturing Δp52&p36 liver stages were infectious for C57BL/6 mice.

  1. Plasmodium berghei Δp52&p36 parasites develop independent of a parasitophorous vacuole membrane in Huh-7 liver cells.

    Science.gov (United States)

    Ploemen, Ivo H J; Croes, Huib J; van Gemert, Geert-Jan J; Wijers-Rouw, Mietske; Hermsen, Cornelus C; Sauerwein, Robert W

    2012-01-01

    The proteins P52 and P36 are expressed in the sporozoite stage of the murine malaria parasite Plasmodium berghei. Δp52&p36 sporozoites lacking expression of both proteins are severely compromised in their capability to develop into liver stage parasites and abort development soon after invasion; presumably due to the absence of a parasitophorous vacuole membrane (PVM). However, a small proportion of P. berghei Δp52&p36 parasites is capable to fully mature in hepatocytes causing breakthrough blood stage infections. We have studied the maturation of replicating Δp52&p36 parasites in cultured Huh-7 hepatocytes. Approximately 50% of Δp52&p36 parasites developed inside the nucleus of the hepatocyte but did not complete maturation and failed to produce merosomes. In contrast cytosolic Δp52&p36 parasites were able to fully mature and produced infectious merozoites. These Δp52&p36 parasites developed into mature schizonts in the absence of an apparent parasitophorous vacuole membrane as shown by immunofluorescence and electron microscopy. Merozoites derived from these maturing Δp52&p36 liver stages were infectious for C57BL/6 mice.

  2. Vacuolating cytotoxin genotypes are strong markers of gastric cancer and duodenal ulcer-associated Helicobacter pylori strains: a matched case-control study.

    Science.gov (United States)

    Memon, Ameer A; Hussein, Nawfal R; Miendje Deyi, Véronique Y; Burette, Alain; Atherton, John C

    2014-08-01

    The Helicobacter pylori virulence gene, cagA, and active forms of the vacuolating cytotoxin gene, vacA, are major determinants of pathogenesis. However, previous studies linking these factors to disease risk have often included patients using aspirin/nonsteroidal anti-inflammatory agents (NSAIDs) or acid-suppressing drugs, both of which may confound results. Also, particularly for gastric cancer (GC), controls have often been of quite different ages. Here, we performed a careful study in a "clean" Belgian population with gastric cancer cases age and sex matched to 4 controls and with a parallel duodenal ulcer (DU) group. As in other populations, there was a close association between the presence of cagA and the vacA s1 genotype. For GC, associations were found for vacA s1-positive (P = 0.01, odds ratio [OR], 9.37; 95% confidence interval [CI], 1.16 to 201.89), i1-positive (P = 0.003; OR, 12.08; 95% CI, 1.50 to 259.64), and cagA-positive status (P ulcer-associated strains are the vacA s1 and i1 genotypes. This fits with experimental data showing that the s and i regions are the key determinants of vacuolating cytotoxin activity.

  3. OsGAP1 functions as a positive regulator of OsRab11-mediated TGN to PM or vacuole trafficking.

    Science.gov (United States)

    Heo, Jae Bok; Rho, Hee Sun; Kim, Se Won; Hwang, Sung Min; Kwon, Hyun Jin; Nahm, Min Yeop; Bang, Woo Young; Bahk, Jeong Dong

    2005-12-01

    The Ypt/Rab family of small G-proteins is important in regulating vesicular transport. Rabs hydrolyze GTP very slowly on their own and require GTPase-activating proteins (GAPs). Here we report the identification and characterization of OsGAP1, a Rab-specific rice GAP. OsGAP1 strongly stimulated OsRab8a and OsRab11, which are homologs of the mammalian Rab8 and Rab11 proteins that are essential for Golgi to plasma membrane (PM) and trans-Golgi network (TGN) to PM trafficking, respectively. Substitution of two invariant arginines within the catalytic domain of Oryza sativa GTPase-activating protein 1 (OsGAP1) with alanines significantly inhibited its GAP activity. In vivo targeting experiments revealed that OsGAP1 localizes to the TGN or pre-vacuolar compartment (PVC). A yeast expression system demonstrated that wild-type OsGAP1 facilitates O. sativa dissociation inhibitor 3 (OsGDI3)-catalyzed OsRab11 recycling at an early stage, but the OsGAP1(R385A) and (R450A) mutants do not. Thus, GTP hydrolysis is essential for Rab recycling. Moreover, expression of the OsGAP1 mutants in Arabidopsis protoplasts inhibited the trafficking of some cargo proteins, including the PM-localizing H+-ATPase-green fluorescent protein (GFP) and Ca2+-ATPase8-GFP and the central vacuole-localizing Arabidopsis aleurain-like protein (AALP)-GFP. The OsGAP1 mutants caused these proteins to accumulate at the Golgi apparatus. Surprisingly, OsRab11 overproduction relieved the inhibitory effect of the OsGAP1 mutants on vesicular trafficking. OsRab8a had no such effect. Thus, the OsGAP1 mutants may inhibit TGN to PM or central vacuole trafficking because they induce the sequestration of endogenous Rab11. We propose that OsGAP1 facilitates vesicular trafficking from the TGN to the PM or central vacuole by both stimulating the GTPase activity of OsRab11 and increasing the recycling of inactive OsRab11.

  4. Phospholipase C-related catalytically inactive protein participates in the autophagic elimination of Staphylococcus aureus infecting mouse embryonic fibroblasts.

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    Kae Harada-Hada

    Full Text Available Autophagy is an intrinsic host defense system that recognizes and eliminates invading bacterial pathogens. We have identified microtubule-associated protein 1 light chain 3 (LC3, a hallmark of autophagy, as a binding partner of phospholipase C-related catalytically inactive protein (PRIP that was originally identified as an inositol trisphosphate-binding protein. Here, we investigated the involvement of PRIP in the autophagic elimination of Staphylococcus aureus in infected mouse embryonic fibroblasts (MEFs. We observed significantly more LC3-positive autophagosome-like vacuoles enclosing an increased number of S. aureus cells in PRIP-deficient MEFs than control MEFs, 3 h and 4.5 h post infection, suggesting that S. aureus proliferates in LC3-positive autophagosome-like vacuoles in PRIP-deficient MEFs. We performed autophagic flux analysis using an mRFP-GFP-tagged LC3 plasmid and found that autophagosome maturation is significantly inhibited in PRIP-deficient MEFs. Furthermore, acidification of autophagosomes was significantly inhibited in PRIP-deficient MEFs compared to the wild-type MEFs, as determined by LysoTracker staining and time-lapse image analysis performed using mRFP-GFP-tagged LC3. Taken together, our data show that PRIP is required for the fusion of S. aureus-containing autophagosome-like vacuoles with lysosomes, indicating that PRIP is a novel modulator in the regulation of the innate immune system in non-professional phagocytic host cells.

  5. Saccharomyces cerevisiae Is Dependent on Vesicular Traffic between the Golgi Apparatus and the Vacuole When Inositolphosphorylceramide Synthase Aur1 Is Inactivated.

    Science.gov (United States)

    Voynova, Natalia S; Roubaty, Carole; Vazquez, Hector M; Mallela, Shamroop K; Ejsing, Christer S; Conzelmann, Andreas

    2015-12-01

    Inositolphosphorylceramide (IPC) and its mannosylated derivatives are the only complex sphingolipids of yeast. Their synthesis can be reduced by aureobasidin A (AbA), which specifically inhibits the IPC synthase Aur1. AbA reportedly, by diminishing IPC levels, causes endoplasmic reticulum (ER) stress, an increase in cytosolic calcium, reactive oxygen production, and mitochondrial damage leading to apoptosis. We found that when Aur1 is gradually depleted by transcriptional downregulation, the accumulation of ceramides becomes a major hindrance to cell survival. Overexpression of the alkaline ceramidase YPC1 rescues cells under this condition. We established hydroxylated C26 fatty acids as a reliable hallmark of ceramide hydrolysis. Such hydrolysis occurs only when YPC1 is overexpressed. In contrast, overexpression of YPC1 has no beneficial effect when Aur1 is acutely repressed by AbA. A high-throughput genetic screen revealed that vesicle-mediated transport between Golgi apparatus, endosomes, and vacuole becomes crucial for survival when Aur1 is repressed, irrespective of the mode of repression. In addition, vacuolar acidification becomes essential when cells are acutely stressed by AbA, and quinacrine uptake into vacuoles shows that AbA activates vacuolar acidification. The antioxidant N-acetylcysteine does not improve cell growth on AbA, indicating that reactive oxygen radicals induced by AbA play a minor role in its toxicity. AbA strongly induces the cell wall integrity pathway, but osmotic support does not improve the viability of wild-type cells on AbA. Altogether, the data support and refine current models of AbA-mediated cell death and add vacuolar protein transport and acidification as novel critical elements of stress resistance.

  6. the versatility of the Helicobacter pylori vacuolating cytotoxin vacA in signal transduction and molecular crosstalk.

    Science.gov (United States)

    Backert, Steffen; Tegtmeyer, Nicole

    2010-01-01

    By modulating important properties of eukaryotic cells, many bacterial protein toxins highjack host signalling pathways to create a suitable niche for the pathogen to colonize and persist. Helicobacter pylori VacA is paradigm of pore-forming toxins which contributes to the pathogenesis of peptic ulceration. Several cellular receptors have been described for VacA, which exert different effects on epithelial and immune cells. The crystal structure of VacA p55 subunit might be important for elucidating details of receptor interaction and pore formation. Here we discuss the multiple signalling activities of this important toxin and the molecular crosstalk between VacA and other virulence factors.

  7. Galaxy formation

    CERN Document Server

    Silk, Joseph; Dvorkin, Irina

    2015-01-01

    Galaxy formation is at the forefront of observation and theory in cosmology. An improved understanding is essential for improving our knowledge both of the cosmological parameters, of the contents of the universe, and of our origins. In these lectures intended for graduate students, galaxy formation theory is reviewed and confronted with recent observational issues. In Lecture 1, the following topics are presented: star formation considerations, including IMF, star formation efficiency and star formation rate, the origin of the galaxy luminosity function, and feedback in dwarf galaxies. In Lecture 2, we describe formation of disks and massive spheroids, including the growth of supermassive black holes, negative feedback in spheroids, the AGN-star formation connection, star formation rates at high redshift and the baryon fraction in galaxies.

  8. Soil formation.

    NARCIS (Netherlands)

    Breemen, van N.; Buurman, P.

    1998-01-01

    Soil Formation deals with qualitative and quantitative aspects of soil formation (or pedogenesis) and the underlying chemical, biological, and physical processes. The starting point of the text is the process - and not soil classification. Effects of weathering and new formation of minerals, mobilis

  9. EhVps32 Is a Vacuole-Associated Protein Involved in Pinocytosis and Phagocytosis of Entamoeaba histolytica.

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    Yunuen Avalos-Padilla

    2015-07-01

    Full Text Available Here, we investigated the role of EhVps32 protein (a member of the endosomal-sorting complex required for transport in endocytosis of Entamoeba histolytica, a professional phagocyte. Confocal microscopy, TEM and cell fractionation revealed EhVps32 in cytoplasmic vesicles and also located adjacent to the plasma membrane. Between 5 to 30 min of phagocytosis, EhVps32 was detected on some erythrocytes-containing phagosomes of acidic nature, and at 60 min it returned to cytoplasmic vesicles and also appeared adjacent to the plasma membrane. TEM images revealed it in membranous structures in the vicinity of ingested erythrocytes. EhVps32, EhADH (an ALIX family member, Gal/GalNac lectin and actin co-localized in the phagocytic cup and in some erythrocytes-containing phagosomes, but EhVps32 was scarcely detected in late phagosomes. During dextran uptake, EhVps32, EhADH and Gal/GalNac lectin, but not actin, co-localized in pinosomes. EhVps32 recombinant protein formed oligomers composed by rings and filaments. Antibodies against EhVps32 monomers stained cytoplasmic vesicles but not erythrocytes-containing phagosomes, suggesting that in vivo oligomers are formed on phagosome membranes. The involvement of EhVps32 in phagocytosis was further study in pNeoEhvps32-HA-transfected trophozoites, which augmented almost twice their rate of erythrophagocytosis as well as the membranous concentric arrays built by filaments, spirals and tunnel-like structures. Some of these structures apparently connected phagosomes with the phagocytic cup. In concordance, the EhVps32-silenced G3 trophozoites ingested 80% less erythrocytes than the G3 strain. Our results suggest that EhVps32 participates in E. histolytica phagocytosis and pinocytosis. It forms oligomers on erythrocytes-containing phagosomes, probably as a part of the scission machinery involved in membrane invagination and intraluminal vesicles formation.

  10. EhVps32 Is a Vacuole-Associated Protein Involved in Pinocytosis and Phagocytosis of Entamoeaba histolytica.

    Science.gov (United States)

    Avalos-Padilla, Yunuen; Betanzos, Abigail; Javier-Reyna, Rosario; García-Rivera, Guillermina; Chávez-Munguía, Bibiana; Lagunes-Guillén, Anel; Ortega, Jaime; Orozco, Esther

    2015-07-01

    Here, we investigated the role of EhVps32 protein (a member of the endosomal-sorting complex required for transport) in endocytosis of Entamoeba histolytica, a professional phagocyte. Confocal microscopy, TEM and cell fractionation revealed EhVps32 in cytoplasmic vesicles and also located adjacent to the plasma membrane. Between 5 to 30 min of phagocytosis, EhVps32 was detected on some erythrocytes-containing phagosomes of acidic nature, and at 60 min it returned to cytoplasmic vesicles and also appeared adjacent to the plasma membrane. TEM images revealed it in membranous structures in the vicinity of ingested erythrocytes. EhVps32, EhADH (an ALIX family member), Gal/GalNac lectin and actin co-localized in the phagocytic cup and in some erythrocytes-containing phagosomes, but EhVps32 was scarcely detected in late phagosomes. During dextran uptake, EhVps32, EhADH and Gal/GalNac lectin, but not actin, co-localized in pinosomes. EhVps32 recombinant protein formed oligomers composed by rings and filaments. Antibodies against EhVps32 monomers stained cytoplasmic vesicles but not erythrocytes-containing phagosomes, suggesting that in vivo oligomers are formed on phagosome membranes. The involvement of EhVps32 in phagocytosis was further study in pNeoEhvps32-HA-transfected trophozoites, which augmented almost twice their rate of erythrophagocytosis as well as the membranous concentric arrays built by filaments, spirals and tunnel-like structures. Some of these structures apparently connected phagosomes with the phagocytic cup. In concordance, the EhVps32-silenced G3 trophozoites ingested 80% less erythrocytes than the G3 strain. Our results suggest that EhVps32 participates in E. histolytica phagocytosis and pinocytosis. It forms oligomers on erythrocytes-containing phagosomes, probably as a part of the scission machinery involved in membrane invagination and intraluminal vesicles formation.

  11. Streptolysin O and its co-toxin NAD-glycohydrolase protect group A Streptococcus from Xenophagic killing.

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    Maghnus O'Seaghdha

    Full Text Available Group A Streptococcus (Streptococcus pyogenes or GAS causes pharyngitis, severe invasive infections, and the post-infectious syndromes of glomerulonephritis and rheumatic fever. GAS can be internalized and killed by epithelial cells in vitro, a process that may contribute to local innate defense against pharyngeal infection. Secretion of the pore-forming toxin streptolysin O (SLO by GAS has been reported to stimulate targeted autophagy (xenophagy upon internalization of the bacteria by epithelial cells. Whereas this process was associated with killing of GAS in HeLa cells, studies in human keratinocytes found SLO production enhanced intracellular survival. To reconcile these conflicting observations, we now report in-depth investigation of xenophagy in response to GAS infection of human oropharyngeal keratinocytes, the predominant cell type of the pharyngeal epithelium. We found that SLO expression was associated with prolonged intracellular survival; unexpectedly, expression of the co-toxin NADase was required for this effect. Enhanced intracellular survival was lost upon deletion of NADase or inactivation of its enzymatic activity. Shortly after internalization of GAS by keratinocytes, SLO-mediated damage to the bacteria-containing vacuole resulted in exposure to the cytosol, ubiquitination of GAS and/or associated vacuolar membrane remnants, and engulfment of GAS in LC3-positive vacuoles. We also found that production of streptolysin S could mediate targeting of GAS to autophagosomes in the absence of SLO, a process accompanied by galectin 8 binding to damaged GAS-containing endosomes. Maturation of GAS-containing autophagosome-like vacuoles to degradative autolysosomes was prevented by SLO pore-formation and by SLO-mediated translocation of enzymatically active NADase into the keratinocyte cytosol. We conclude that SLO stimulates xenophagy in pharyngeal keratinocytes, but the coordinated action of SLO and NADase prevent maturation of GAS

  12. Excessive L-cysteine induces vacuole-like cell death by activating endoplasmic reticulum stress and mitogen-activated protein kinase signaling in intestinal porcine epithelial cells.

    Science.gov (United States)

    Ji, Yun; Wu, Zhenlong; Dai, Zhaolai; Sun, Kaiji; Zhang, Qing; Wu, Guoyao

    2016-01-01

    High intake of dietary cysteine is extremely toxic to animals and the underlying mechanism remains largely unknown. This study was conducted to test the hypothesis that excessive L-cysteine induces cell death by activating endoplasmic reticulum (ER) stress and mitogen-activated protein kinase (MAPK) signaling in intestinal porcine epithelial cells. Jejunal enterocytes were cultured in the presence of 0-10 mmol/L L-cysteine. Cell viability, morphologic alterations, mRNA levels for genes involved in ER stress, protein abundances for glucose-regulated protein 78, C/EBP homologous protein (CHOP), alpha subunit of eukaryotic initiation factor-2 (eIF2α), extracellular signal-regulated kinase (ERK1/2), p38 MAPK, and c-Jun N-terminal protein kinase (JNK1/2) were determined. The results showed that L-cysteine (5-10 mmol/L) reduced cell viability (P cysteine were not affected by the autophagy inhibitor 3-methyladenine. The protein abundances for CHOP, phosphorylated (p)-eIF2α, p-JNK1/2, p-p38 MAPK, and the spliced form of XBP-1 mRNA were enhanced (P cysteine induces vacuole-like cell death via the activation of ER stress and MAPK signaling in small intestinal epithelial cells. These signaling pathways may be potential targets for developing effective strategies to prevent the toxicity of dietary cysteine.

  13. Structure of Yeast OSBP-Related Protein Osh1 Reveals Key Determinants for Lipid Transport and Protein Targeting at the Nucleus-Vacuole Junction.

    Science.gov (United States)

    Manik, Mohammad Kawsar; Yang, Huiseon; Tong, Junsen; Im, Young Jun

    2017-03-10

    Yeast Osh1 belongs to the oxysterol-binding protein (OSBP) family of proteins and contains multiple targeting modules optimized for lipid transport at the nucleus-vacuole junction (NVJ). The key determinants for NVJ targeting and the role of Osh1 at NVJs have remained elusive because of unknown lipid specificities. In this study, we determined the structures of the ankyrin repeat domain (ANK), and OSBP-related domain (ORD) of Osh1, in complex with Nvj1 and ergosterol, respectively. The Osh1 ANK forms a unique bi-lobed structure that recognizes a cytosolic helical segment of Nvj1. We discovered that Osh1 ORD binds ergosterol and phosphatidylinositol 4-phosphate PI(4)P in a competitive manner, suggesting counter-transport function of the two lipids. Ergosterol is bound to the hydrophobic pocket in a head-down orientation, and the structure of the PI(4)P-binding site in Osh1 is well conserved. Our results suggest that Osh1 performs non-vesicular transport of ergosterol and PI(4)P at the NVJ.

  14. The E2-like conjugation enzyme Atg3 promotes binding of IRG and Gbp proteins to Chlamydia- and Toxoplasma-containing vacuoles and host resistance.

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    Arun K Haldar

    Full Text Available Cell-autonomous immunity to the bacterial pathogen Chlamydia trachomatis and the protozoan pathogen Toxoplasma gondii is controlled by two families of Interferon (IFN-inducible GTPases: Immunity Related GTPases (IRGs and Guanylate binding proteins (Gbps. Members of these two GTPase families associate with pathogen-containing vacuoles (PVs and solicit antimicrobial resistance pathways specifically to the intracellular site of infection. The proper delivery of IRG and Gbp proteins to PVs requires the autophagy factor Atg5. Atg5 is part of a protein complex that facilitates the transfer of the ubiquitin-like protein Atg8 from the E2-like conjugation enzyme Atg3 to the lipid phosphatidylethanolamine. Here, we show that Atg3 expression, similar to Atg5 expression, is required for IRG and Gbp proteins to dock to PVs. We further demonstrate that expression of a dominant-active, GTP-locked IRG protein variant rescues the PV targeting defect of Atg3- and Atg5-deficient cells, suggesting a possible role for Atg proteins in the activation of IRG proteins. Lastly, we show that IFN-induced cell-autonomous resistance to C. trachomatis infections in mouse cells depends not only on Atg5 and IRG proteins, as previously demonstrated, but also requires the expression of Atg3 and Gbp proteins. These findings provide a foundation for a better understanding of IRG- and Gbp-dependent cell-autonomous resistance and its regulation by Atg proteins.

  15. A RAB3GAP1 SINE Insertion in Alaskan Huskies with Polyneuropathy, Ocular Abnormalities, and Neuronal Vacuolation (POANV Resembling Human Warburg Micro Syndrome 1 (WARBM1

    Directory of Open Access Journals (Sweden)

    Michaela Wiedmer

    2016-02-01

    Full Text Available We observed a hereditary phenotype in Alaskan Huskies that was characterized by polyneuropathy with ocular abnormalities and neuronal vacuolation (POANV. The affected dogs developed a progressive severe ataxia, which led to euthanasia between 8 and 16 months of age. The pedigrees were consistent with a monogenic autosomal recessive inheritance. We localized the causative genetic defect to a 4 Mb interval on chromosome 19 by a combined linkage and homozygosity mapping approach. Whole genome sequencing of one affected dog, an obligate carrier, and an unrelated control revealed a 218-bp SINE insertion into exon 7 of the RAB3GAP1 gene. The SINE insertion was perfectly associated with the disease phenotype in a cohort of 43 Alaskan Huskies, and it was absent from 541 control dogs of diverse other breeds. The SINE insertion induced aberrant splicing and led to a transcript with a greatly altered exon 7. RAB3GAP1 loss-of-function variants in humans cause Warburg Micro Syndrome 1 (WARBM1, which is characterized by additional developmental defects compared to canine POANV, whereas Rab3gap1-deficient mice have a much milder phenotype than either humans or dogs. Thus, the RAB3GAP1 mutant Alaskan Huskies provide an interesting intermediate phenotype that may help to better understand the function of RAB3GAP1 in development. Furthermore, the identification of the presumed causative genetic variant will enable genetic testing to avoid the nonintentional breeding of affected dogs.

  16. Molecular cloning, immunochemical localization to the vacuole, and expression in transgenic yeast and tobacco of a putative sugar transporter from sugar beet.

    Science.gov (United States)

    Chiou, T J; Bush, D R

    1996-02-01

    Several plant genes have been cloned that encode members of the sugar transporter subgroup of the major facilitator superfamily of transporters. Here we report the cloning, expression, and membrane localization of one of these porters found in sugar beet (Beta vulgaris L.). This clone, cDNA-1, codes for a protein with 490 amino acids and an estimated molecular mass of 54 kD. The predicted membrane topology and sequence homology suggest that cDNA-1 is a member of the sugar transporter family. RNA gel blot analysis revealed that this putative sugar transporter is expressed in all vegetative tissues and expression increases with development in leaves. DNA gel blot analysis indicated that multiple gene copies exist for this putative sugar transporter in the sugar beet genome. Antibodies directed against small peptides representing the N- and C-terminal domains of the cDNA1 protein identified a 40-kD polypeptide in microsomes isolated from cDNA-1-transformed yeast (Saccharomyces cerevisiae). Moreover, the same protein was identified in sugar beet and transgenic tobacco (Nicotaina tobacum L.) membrane fractions. Detailed analysis of the transporter's distribution across linear sucrose gradients and flotation centrifugations showed that it co-migrates with tonoplast membrane markers. We conclude that this carrier is located on the tonoplast membrane and that it may mediate sugar partitioning between the vacuole and cytoplasmic compartments.

  17. cPrG-HCl a potential H+/Cl- symporter prevents acidification of storage vacuoles in aleurone cells and inhibits GA-dependent hydrolysis of storage protein and phytate.

    Science.gov (United States)

    Hwang, Yong-sic; Bethke, Paul C; Gubler, Frank; Jones, Russell L

    2003-07-01

    The putative H+/Cl- symporter cycloprodigiosin-HCl (cPrG-HCl) was used to investigate the role of vacuole acidification in cereal aleurone cell function. The protein storage vacuole (PSV) becomes acidified rapidly when aleurone cells are treated with gibberellic acid (GA) but not abscisic acid (ABA). We show that cPrG prevents PSV acidification in aleurone layers and prevents synthesis of secretory proteins such as alpha-amylase. Our data support the hypothesis that decreased hydrolase synthesis is a consequence of decreased hydrolysis of storage proteins in PSV. Support for this hypothesis comes from experiments showing that breakdown of barley 7S globulins and phytate is inhibited by cPrG in GA-treated aleurone layers. Decreased mobilization of PSV reserves is accompanied by reductions in the free amino acid pool size and in the amount of ions released from the aleurone layer. Vacuolation of the aleurone cell is a diagnostic feature of the response to GA, and vacuolation is also inhibited by cPrG. Evidence that cPrG acts as a potential H+/Cl- symporter in aleurone is presented. We show that cPrG does not inhibit the synthesis and secretion of alpha-amylase when Cl- ions are omitted from the incubation medium. Although cPrG blocks many GA-induced responses of aleurone layers, it does not affect early steps in GA signaling. The SLN1 protein, a negative regulator of GA signaling, is turned over in GA-treated cells in the presence and absence of cPrG. Similarly, synthesis of the transcriptional activator GAMYB is unaffected by the presence of cPrG in GA-treated cells.

  18. Expression of a vacuole-localized BURP-domain protein from soybean (SALI3-2 enhances tolerance to cadmium and copper stresses.

    Directory of Open Access Journals (Sweden)

    Yulin Tang

    Full Text Available The plant-specific BURP family proteins play diverse roles in plant development and stress responses, but the function mechanism of these proteins is still poorly understood. Proteins in this family are characterized by a highly conserved BURP domain with four conserved Cys-His repeats and two other Cys, indicating that these proteins potentially interacts with metal ions. In this paper, an immobilized metal affinity chromatography (IMAC assay showed that the soybean BURP protein SALI3-2 could bind soft transition metal ions (Cd(2+, Co(2+, Ni(2+, Zn(2+ and Cu(2+ but not hard metal ions (Ca(2+ and Mg(2+ in vitro. A subcellular localization analysis by confocal laser scanning microscopy revealed that the SALI3-2-GFP fusion protein was localized to the vacuoles. Physiological indexes assay showed that Sali3-2-transgenic Arabidopsis thaliana seedlings were more tolerant to Cu(2+ or Cd(2+ stresses than the wild type. An inductively coupled plasma optical emission spectrometry (ICP-OES analysis illustrated that, compared to the wild type seedlings the Sali3-2-transgenic seedlings accumulated more cadmium or copper in the roots but less in the upper ground tissues when the seedlings were exposed to excessive CuCl2 or CdCl2 stress. Therefore, our findings suggest that the SALI3-2 protein may confer cadmium (Cd(2+ and copper (Cu(2+ tolerance to plants by helping plants to sequester Cd(2+ or Cu(2+ in the root and reduce the amount of heavy metals transported to the shoots.

  19. Salmonella Effectors SseF and SseG Interact with Mammalian Protein ACBD3 (GCP60 To Anchor Salmonella-Containing Vacuoles at the Golgi Network

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    Xiu-Jun Yu

    2016-07-01

    Full Text Available Following infection of mammalian cells, Salmonella enterica serovar Typhimurium (S. Typhimurium replicates within membrane-bound compartments known as Salmonella-containing vacuoles (SCVs. The Salmonella pathogenicity island 2 type III secretion system (SPI-2 T3SS translocates approximately 30 different effectors across the vacuolar membrane. SseF and SseG are two such effectors that are required for SCVs to localize close to the Golgi network in infected epithelial cells. In a yeast two-hybrid assay, SseG and an N-terminal variant of SseF interacted directly with mammalian ACBD3, a multifunctional cytosolic Golgi network-associated protein. Knockdown of ACBD3 by small interfering RNA (siRNA reduced epithelial cell Golgi network association of wild-type bacteria, phenocopying the effect of null mutations of sseG or sseF. Binding of SseF to ACBD3 in infected cells required the presence of SseG. A single-amino-acid mutant of SseG and a double-amino-acid mutant of SseF were obtained that did not interact with ACBD3 in Saccharomyces cerevisiae. When either of these was produced together with the corresponding wild-type effector by Salmonella in infected cells, they enabled SCV-Golgi network association and interacted with ACBD3. However, these properties were lost and bacteria displayed an intracellular replication defect when cells were infected with Salmonella carrying both mutant genes. Knockdown of ACBD3 resulted in a replication defect of wild-type bacteria but did not further attenuate the growth defect of a ΔsseFG mutant strain. We propose a model in which interaction between SseF and SseG enables both proteins to bind ACBD3, thereby anchoring SCVs at the Golgi network and facilitating bacterial replication.

  20. Galaxy Formation

    DEFF Research Database (Denmark)

    Sparre, Martin

    Galaxy formation is an enormously complex discipline due to the many physical processes that play a role in shaping galaxies. The objective of this thesis is to study galaxy formation with two different approaches: First, numerical simulations are used to study the structure of dark matter and how...... galaxies form stars throughout the history of the Universe, and secondly it is shown that observations of gamma-ray bursts (GRBs) can be used to probe galaxies with active star formation in the early Universe. A conclusion from the hydrodynamical simulations is that the galaxies from the stateof......-the-art cosmological simulation, Illustris, follow a tight relation between star formation rate and stellar mass. This relation agrees well with the observed relation at a redshift of z = 0 and z = 4, but at intermediate redshifts of z ' 2 the normalisation is lower than in real observations. This is highlighted...

  1. Galaxy Formation

    DEFF Research Database (Denmark)

    Sparre, Martin

    galaxies form stars throughout the history of the Universe, and secondly it is shown that observations of gamma-ray bursts (GRBs) can be used to probe galaxies with active star formation in the early Universe. A conclusion from the hydrodynamical simulations is that the galaxies from the stateof......-the-art cosmological simulation, Illustris, follow a tight relation between star formation rate and stellar mass. This relation agrees well with the observed relation at a redshift of z = 0 and z = 4, but at intermediate redshifts of z ' 2 the normalisation is lower than in real observations. This is highlighted...... of GRB host galaxies is affected by the fact that GRBs appear mainly to happen in low-metallicity galaxies. Solving this problem will make it possible to derive the total cosmic star formation rate more reliably from number counts of GRBs....

  2. The pesticide methoxychlor decreases myotube formation in cell culture by slowing myoblast proliferation.

    Science.gov (United States)

    Steffens, Bradley W; Batia, Lyn M; Baarson, Chad J; Choi, Chang-Kun Charles; Grow, Wade A

    2007-08-01

    We studied the effect of the estrogenic pesticide methoxychlor (MXC) on skeletal muscle development using C2C12 cell culture. Myoblast cultures were exposed to various concentrations of MXC at various times during the process of myoblast fusion into myotubes. We observed that MXC exposure decreased myotube formation. In addition, we observed myoblasts with cytoplasmic vacuoles in cultures exposed to MXC. Because cytoplasmic vacuoles can be characteristic of cell death, apoptosis assays and trypan blue exclusion assays were performed. We found no difference in the frequency of apoptosis or in the frequency of cell death for cultures exposed to MXC and untreated cultures. Collectively, these results indicate that MXC exposure decreases myotube formation without causing cell death. In contrast, when cell proliferation was assessed, untreated cultures had a myoblast proliferation rate 50% greater than cultures exposed to MXC. We conclude that MXC decreases myotube formation at least in part by slowing myoblast proliferation. Furthermore, we suggest that direct exposure to MXC could affect skeletal muscle development in animals or humans, in addition to the defects in reproductive development that have previously been reported.

  3. Hippocampal formation

    NARCIS (Netherlands)

    Cappaert, N.L.M.; van Strien, N.M.; Witter, M.P.; Paxinos, G.

    2015-01-01

    The hippocampal formation and parahippocampal region are prominent components of the rat nervous system and play a crucial role in learning, memory, and spatial navigation. Many new details regarding the entorhinal cortex have been discovered since the previous edition, and the growing interest in t

  4. Galaxy formation.

    Science.gov (United States)

    Peebles, P J

    1998-01-01

    It is argued that within the standard Big Bang cosmological model the bulk of the mass of the luminous parts of the large galaxies likely had been assembled by redshift z approximately 10. Galaxy assembly this early would be difficult to fit in the widely discussed adiabatic cold dark matter model for structure formation, but it could agree with an isocurvature version in which the cold dark matter is the remnant of a massive scalar field frozen (or squeezed) from quantum fluctuations during inflation. The squeezed field fluctuations would be Gaussian with zero mean, and the distribution of the field mass therefore would be the square of a random Gaussian process. This offers a possibly interesting new direction for the numerical exploration of models for cosmic structure formation.

  5. Cloud Formation

    Science.gov (United States)

    Graham, Mark Talmage

    2004-05-01

    Cloud formation is crucial to the heritage of modern physics, and there is a rich literature on this important topic. In 1927, Charles T.R. Wilson was awarded the Nobel Prize in physics for applications of the cloud chamber.2 Wilson was inspired to study cloud formation after working at a meteorological observatory on top of the highest mountain in Scotland, Ben Nevis, and testified near the end of his life, "The whole of my scientific work undoubtedly developed from the experiments I was led to make by what I saw during my fortnight on Ben Nevis in September 1894."3 To form clouds, Wilson used the sudden expansion of humid air.4 Any structure the cloud may have is spoiled by turbulence in the sudden expansion, but in 1912 Wilson got ion tracks to show up by using strobe photography of the chamber immediately upon expansion.5 In the interim, Millikan's study in 1909 of the formation of cloud droplets around individual ions was the first in which the electron charge was isolated. This study led to his famous oil drop experiment.6 To Millikan, as to Wilson, meteorology and physics were professionally indistinct. With his meteorological physics expertise, in WWI Millikan commanded perhaps the first meteorological observation and forecasting team essential to military operation in history.7 But even during peacetime meteorology is so much of a concern to everyone that a regular news segment is dedicated to it. Weather is the universal conversation topic, and life on land could not exist as we know it without clouds. One wonders then, why cloud formation is never covered in physics texts.

  6. The formation of Anthocyanic Vacuolar Inclusions in Arabidopsis thaliana and implications for the sequestration of anthocyanin pigments.

    Science.gov (United States)

    Pourcel, Lucille; Irani, Niloufer G; Lu, Yuhua; Riedl, Ken; Schwartz, Steve; Grotewold, Erich

    2010-01-01

    Anthocyanins are flavonoid pigments that accumulate in the large central vacuole of most plants. Inside the vacuole, anthocyanins can be found uniformly distributed or as part of sub-vacuolar pigment bodies, the Anthocyanic Vacuolar Inclusions (AVIs). Using Arabidopsis seedlings grown under anthocyanin-inductive conditions as a model to understand how AVIs are formed, we show here that the accumulation of AVIs strongly correlates with the formation of cyanidin 3-glucoside (C3G) and derivatives. Arabidopsis mutants that fail to glycosylate anthocyanidins at the 5-O position (5gt mutant) accumulate AVIs in almost every epidermal cell of the cotyledons, as compared to wild-type seedlings, where only a small fraction of the cells show AVIs. A similar phenomenon is observed when seedlings are treated with vanadate. Highlighting a role for autophagy in the formation of the AVIs, we show that various mutants that interfere with the autophagic process (atg mutants) display lower numbers of AVIs, in addition to a reduced accumulation of anthocyanins. Interestingly, vanadate increases the numbers of AVIs in the atg mutants, suggesting that several pathways might participate in AVI formation. Taken together, our results suggest novel mechanisms for the formation of sub-vacuolar compartments capable of accumulating anthocyanin pigments.

  7. Cement Formation

    DEFF Research Database (Denmark)

    Telschow, Samira; Jappe Frandsen, Flemming; Theisen, Kirsten

    2012-01-01

    Cement production has been subject to several technological changes, each of which requires detailed knowledge about the high multiplicity of processes, especially the high temperature process involved in the rotary kiln. This article gives an introduction to the topic of cement, including...... an overview of cement production, selected cement properties, and clinker phase relations. An extended summary of laboratory-scale investigations on clinkerization reactions, the most important reactions in cement production, is provided. Clinker formations by solid state reactions, solid−liquid and liquid...

  8. Vacuolating encephalopathy and retinopathy associated with a nodavirus-like agent: a probable cause of mass mortality of wild Golden grey mullet (Liza aurata) and Sharpnose grey mullet (Liza saliens) in Iranian waters of the Caspian Sea.

    Science.gov (United States)

    Zorriehzahra, Mohammad Jalil; Nazari, Alireza; Ghasemi, Mohaddes; Ghiasi, Maryam; Karsidani, Somayeh Haghighi; Bovo, Giuseppe; Daud, Hassan Hj Mohd

    2014-12-01

    Mullets are dominant fishes in the catch composition in the southern coasts of the Caspian Sea and after (Rutilus frisii kutum Kamensky, 1901) have a worthy role in production of marine proteins and incomings of north provinces of Iran. Mullets stocks decreased dramatically in recent decades in the Caspian Sea and catch amount reached from 6446 MT on 2002 to 2151 MT in 2012. Mysterious mortalities occurred in wild mullet (Liza auratu) and (Liza saliens) in Iranian waters of Caspian Sea in recent years. Regarding to investigation of causative agent of mentioned outbreak about 322 suspected samples were collected from coastal capture sites of Iranian north provinces in 2008 till 2011. Moribund fish revealed skin darkening, erratic swimming, belly-up at rest and high distension of swim bladder. Target tissues such as brain and eye were removed and then fixed for histopathology and TEM assay. Widespread and massive vacuolation were observed in brain, spinal cord, retina and optical nerve and intracytoplasmic vacuoles and virus particles in retina. So concerning to clinical signs, histopathological and TEM findings, it could be concluded that nodavirus-like agent could be probable cause of mass mortality of wild mullet in Iranian waters of the Caspian Sea.

  9. Galaxy Formation

    CERN Document Server

    Longair, Malcolm S

    2008-01-01

    This second edition of Galaxy Formation is an up-to-date text on astrophysical cosmology, expounding the structure of the classical cosmological models from a contemporary viewpoint. This forms the background to a detailed study of the origin of structure and galaxies in the Universe. The derivations of many of the most important results are derived by simple physical arguments which illuminate the results of more advanced treatments. A very wide range of observational data is brought to bear upon these problems, including the most recent results from WMAP, the Hubble Space Telescope, galaxy surveys like the Sloan Digital Sky Survey and the 2dF Galaxy Redshift Survey, studies of Type 1a supernovae, and many other observations.

  10. Pattern Formation

    Science.gov (United States)

    Hoyle, Rebecca

    2006-03-01

    From the stripes of a zebra and the spots on a leopard's back to the ripples on a sandy beach or desert dune, regular patterns arise everywhere in nature. The appearance and evolution of these phenomena has been a focus of recent research activity across several disciplines. This book provides an introduction to the range of mathematical theory and methods used to analyse and explain these often intricate and beautiful patterns. Bringing together several different approaches, from group theoretic methods to envelope equations and theory of patterns in large-aspect ratio-systems, the book also provides insight behind the selection of one pattern over another. Suitable as an upper-undergraduate textbook for mathematics students or as a fascinating, engaging, and fully illustrated resource for readers in physics and biology, Rebecca Hoyle's book, using a non-partisan approach, unifies a range of techniques used by active researchers in this growing field. Accessible description of the mathematical theory behind fascinating pattern formation in areas such as biology, physics and materials science Collects recent research for the first time in an upper level textbook Features a number of exercises - with solutions online - and worked examples

  11. 咪唑诱导的人子宫内膜癌细胞HEC-1B空泡化现象研究%Study on the Vacuolation of Imidazole-Treated Human Endometrial Carcinoma Cell Line HEC-1B

    Institute of Scientific and Technical Information of China (English)

    张玖航; 曾申明

    2011-01-01

    As a kind of weak bases, imidazole cause vacuolation of animal endometrial cells, and affect the physiological and reproductive functions. Our study discuss the mechanism of vacuolation after treatment with imidazole through experimental model of human endometrial carcinoma cell line HEC -IB. We study through Genechip research, fluorescence quantitative PCR and observation of laser confocal microscopy. Results: Genechip research results show that gene of subunit V0D2 of human vacuolar type H+-ATPase (V-ATPase) had an increased expression after the treatment with imidazole on human endometrial carcinoma cell line HEC -IB. Genechip research results were verified by fluorescence quantitative PCR. The expression of the mRNA of subunit V0D2 of human V -ATPase increased.Another subunit V1E1 of human V-ATPase also had an increased expression by fluorescence quantitative PCR. Bafilomycin Al, an inhibitor of V-ATPase, significantly inhibited the vacuolation caused by imidazole. The phenomenon was observed by laser confocal microscopy after staining human endometrial carcinoma cell line HEC-IB with Acridine orange. These results suggest that V-ATPase plays an important role in cell vacuolation caused by imidazole.%咪唑作为一种弱碱性物质,处理动物子宫内膜细胞后,可诱导其产生空泡化现象,进而影响其生殖生理功能.本研究使用人子宫内膜癌细胞HEC-1B作为模型,使用咪唑处理导致细胞空泡化后探讨空泡化产生的生理机制.通过基因芯片分析、荧光定量PCR检测、激光共聚焦显微镜观察等方法进行研究.结果表明:咪唑处理人子宫内膜癌细胞HEC-1B后,空泡型质子泵(V-ATPase)亚基VOD2基因表达量上调.使用荧光定量PCR验证芯片结果,V-ATPase亚基VOD2mRNA表达量有所上调.对V-ATPase另外一个亚基V1E1基因进行荧光定量PCR,其mRNA表达量也有所上调.使用V-ATPase抑制剂巴弗洛霉素A1和咪唑共同处理人子宫内膜癌细胞HEC-1B后,通过

  12. Structures in material transference and vitelline envelope formation in Betta splendens follicles.

    Science.gov (United States)

    Genta, H D

    1996-01-01

    Structures were found by transmission electron microscopy, they were located within follicular cells and the oocyte, and in the interspace between them in follicles of the teleost fish Betta splendens. Some structures with features characteristic or lamellar bodies were found in small follicles. The possible role of these structures in the formation of the vitelline envelope as well as in the material transference is discussed. Vacuoles, vesticles and particles intensely stained were found in the microvilli and the cortical cytoplasm of the oocyte at the onset of vitellogenesis. These results suggest that different substances present in the cellular components of the follicle might be transferred from cell to cell through the extracellular space and through the prolongations that cross the extracellular space.

  13. Guanine nucleotide exchange factor 2 for Rab5 proteins coordinated with GLUP6/GEF regulates the intracellular transport of the proglutelin from the Golgi apparatus to the protein storage vacuole in rice endosperm.

    Science.gov (United States)

    Wen, Liuying; Fukuda, Masako; Sunada, Mariko; Ishino, Sonoko; Ishino, Yoshizumi; Okita, Thomas W; Ogawa, Masahiro; Ueda, Takashi; Kumamaru, Toshihiro

    2015-10-01

    Rice glutelin polypeptides are initially synthesized on the endoplasmic reticulum (ER) membrane as a proglutelin, which are then transported to the protein storage vacuole (PSV) via the Golgi apparatus. Rab5 and its cognate activator guanine nucleotide exchange factor (GEF) are essential for the intracellular transport of proglutelin from the Golgi apparatus to the PSV. Results from previous studies showed that the double recessive type of glup4/rab5a and glup6/gef mutant accumulated much higher amounts of proglutelin than either parent line. The present study demonstrates that the double recessive type of glup4/rab5a and glup6/gef mutant showed not only elevated proglutelin levels and much larger paramural bodies but also reduced the number and size of PSVs, indicating a synergistic mutation effect. These observations led us to the hypothesis that other isoforms of Rab5 and GEF also participate in the intracellular transport of rice glutelin. A database search identified a novel guanine nucleotide exchange factor, Rab5-GEF2. Like GLUP6/GEF, Rab5-GEF2 was capable of activating Rab5a and two other Rab5 isoforms in in vitro GTP/GDP exchange assays. GEF proteins consist of the helical bundle (HB) domain at the N-terminus, Vps9 domain, and a C-terminal region. By the deletion analysis of GEFs, the HB domain was found essential for the activation of Rab5 proteins.

  14. Symbiotic Chlorella variabilis incubated under constant dark conditions for 24 hours loses the ability to avoid digestion by host lysosomal enzymes in digestive vacuoles of host ciliate Paramecium bursaria.

    Science.gov (United States)

    Kodama, Yuuki; Fujishima, Masahiro

    2014-12-01

    Endosymbiosis between symbiotic Chlorella and alga-free Paramecium bursaria cells can be induced by mixing them. To establish the endosymbiosis, algae must acquire temporary resistance to the host lysosomal enzymes in the digestive vacuoles (DVs). When symbiotic algae isolated from the alga-bearing paramecia are kept under a constant dark conditions for 24 h before mixing with the alga-free paramecia, almost all algae are digested in the host DVs. To examine the cause of algal acquisition to the host lysosomal enzymes, the isolated algae were kept under a constant light conditions with or without a photosynthesis inhibitor 3-(3,4-dichlorophenyl)-1,1-dimethylurea for 24 h, and were mixed with alga-free paramecia. Unexpectedly, most of the algae were not digested in the DVs irrespective of the presence of the inhibitor. Addition of 1 mM maltose, a main photosynthetic product of the symbiotic algae or of a supernatant of the isolated algae kept for 24 h under a constant light conditions, did not rescue the algal digestion in the DVs. These observations reveal that unknown factors induced by light are a prerequisite for algal resistance to the host lysosomal enzymes.

  15. THE EXPRESSION OF VACA IN THE BCF OF HELICOBACTER PYLORI AND ITS RELATIONSHIP TO THE VACUOLATED EFFECT%VacA在幽门螺杆菌BCF中的表达及其与空泡毒作用的关系

    Institute of Scientific and Technical Information of China (English)

    施理; 侯晓华; 易粹琼

    2002-01-01

    目的:研究幽门螺杆菌(Helicobacter pylori,H.pylori)临床分离株的空泡毒作用及其与空泡毒素抗原(Vacuolated cytotoxin antigen,VacA)的关系.方法:用细胞毒试验、SDS-PAGE结合薄层扫描研究H.pylori临床分离株空泡毒作用及其与VacA的关系.结果:62株H.pylori临床分离株,43株为空泡毒作用阳性 H.pylori(Toxin+) ,其余为空泡毒作用阴性的H.pylori(Toxin-).30.23%(13/43)H.pylori(Toxin+)临床分离株的肉汤培养滤液(broth culture filter,BCF)含有分子量为87 kDa的VacA,而所有H.pylori(Toxin-)的BCF不含有这种蛋白,二者之间差异有显著性(P<0.05);VacA含量与H.pylori(Toxin+)BCF的空泡毒作用滴度明显相关(r=0.67,P<0.05).结论:H.pylori(Toxin+)空泡毒作用是由VacA引起的.

  16. Common File Formats.

    Science.gov (United States)

    Mills, Lauren

    2014-03-21

    An overview of the many file formats commonly used in bioinformatics and genome sequence analysis is presented, including various data file formats, alignment file formats, and annotation file formats. Example workflows illustrate how some of the different file types are typically used.

  17. Culture supernatants from V. cholerae O1 ElTor strains isolated from different geographic areas induce cell vacuolation and cytotoxicity Cepas de V. cholerae O1 biotipo ElTor aisladas de diferente origen geográfico inducen vacuolización celular y citotoxicidad

    Directory of Open Access Journals (Sweden)

    Jorge E Vidal

    2009-02-01

    Full Text Available OBJECTIVE: To investigate whether the HlyA-induced vacuolating effect is produced by V. cholerae O1 ElTor strains isolated from different geographic origins, including Mexico. MATERIAL AND METHODS: Supernatant-induced haemolysis, vacuolating activity and cytotoxicity in Vero cells were recorded. PCR, RFLP analysis and molecular cloning were performed. RESULTS: All ElTor strains analyzed induced cellular vacuolation. Ribotype 2 strains isolates from the U.S. gulf coast yielded the highest titer of vacuolating activity. Eight of nine strains were haemolytic, while all strains were PCR positive for the hlyA gene. We cloned the hlyA gene from two ElTor strains, a toxigenic (2514-88, ctxAB+ and a non-toxigenic Mexican strain (CM 91-3, ctxAB-. Supernatant from those recombinant E. coli strains induced haemolysis, cell vacuolation and cytotoxicity. RFLP-PCR analysis revealed similarities in the hlyA gene from all strains tested. CONCLUSION: The HlyA-induced vacuolating effect is a widespread phenotype of epidemic V. cholerae O1 ElTor strains.OBJETIVO: Analizar el efecto vacuolizante de cepas de V. cholerae O1 ElTor aisladas de diferente origen geográfico, incluyendo México. MATERIAL Y MÉTODOS: Se realizaron pruebas de hemolisis, vacuolización y citotoxicidad en células Vero, así como PCR, análisis por RFLP y clonación molecular. RESULTADOS: Todas las cepas indujeron el efecto vacuolizante. Las cepas del ribotipo 2, aisladas de las costas del Golfo en Estados Unidos, presentaron títulos altos de vacuolización. El gen hlyA fue amplificado en las nueve cepas mediante PCR, aunque sólo ocho fueron hemolíticas. Se clonó el gen hlyA de una cepa toxigénica (2514-88, ctxAB+ y de una cepa no toxigénica aislada en México (CM 91-3, ctxAB-. El sobrenadante de las clonas recombinantes indujo hemólisis, efecto vacuolizante y citotoxicidad. El RFLP mostró alta similitud del gen hlyA de las cepas estudiadas. CONCLUSIÓN: El efecto vacuolizante es un

  18. The Format Registry Problem

    Directory of Open Access Journals (Sweden)

    Gary McGath

    2013-01-01

    Full Text Available File format identification is an important issue in digital preservation. Several noteworthy attempts, including PRONOM, GDFR, and UDFR, have been made at creating a comprehensive repository of format information. The sheer amount of information to cover and the constant introduction of new formats and format versions has limited their success. Alternative approaches, such as Linked Data and offering limited per-format information with identifiers that can be used elsewhere, may lead to greater success.

  19. Formate Formation and Formate Conversion in Biological Fuels Production

    Directory of Open Access Journals (Sweden)

    Bryan R. Crable

    2011-01-01

    Full Text Available Biomethanation is a mature technology for fuel production. Fourth generation biofuels research will focus on sequestering CO2 and providing carbon-neutral or carbon-negative strategies to cope with dwindling fossil fuel supplies and environmental impact. Formate is an important intermediate in the methanogenic breakdown of complex organic material and serves as an important precursor for biological fuels production in the form of methane, hydrogen, and potentially methanol. Formate is produced by either CoA-dependent cleavage of pyruvate or enzymatic reduction of CO2 in an NADH- or ferredoxin-dependent manner. Formate is consumed through oxidation to CO2 and H2 or can be further reduced via the Wood-Ljungdahl pathway for carbon fixation or industrially for the production of methanol. Here, we review the enzymes involved in the interconversion of formate and discuss potential applications for biofuels production.

  20. Polaribacter gen. nov., with three new species, P. irgensii sp. nov., P. franzmannii sp. nov. and P. filamentus sp. nov., gas vacuolate polar marine bacteria of the Cytophaga-Flavobacterium-Bacteroides group and reclassification of 'Flectobacillus glomeratus' as Polaribacter glomeratus comb. nov

    Science.gov (United States)

    Gosink, J. J.; Woese, C. R.; Staley, J. T.

    1998-01-01

    Several psychrophilic, gas vacuolate strains of the Cytophage-Flavobacterium-Bacteroides (CFB) phylogenetic group were isolated from sea ice and water from the Arctic and the Antarctic. The closest taxonomically defined species by 16S rRNA sequence analysis is 'Flectobacillus glomeratus'. However, 'Flc. glomeratus' is phylogenetically distant from the Flectobacillus type species, Flc. major. On the basis of phenotypic, genotypic and 16S rRNA sequence analyses we propose a new genus, Polaribacter, with three new species, Polaribacter irgensii strain 23-P (ATCC 700398), Polaribacter franzmannii strain 301 (ATCC 700399) and Polaribacter filamentus strain 215 (ATCC 700397). P. filamentus is the type species of the genus. None of these species exhibits a cosmopolitan or bipolar distribution. This is the first taxonomic description of gas vacuolate bacteria in the CFB group. Additionally, we propose that 'Flc. glomeratus' be reclassified to the genus Polaribacter as P. glomeratus, comb. nov.

  1. Toxic response of HIPCO single-walled carbon nanotubes in mice and RAW264.7 macrophage cells.

    Science.gov (United States)

    Park, Eun-Jung; Zahari, Nur Elida M; Kang, Min-Sung; Lee, Sang jin; Lee, Kyuhong; Lee, Byoung-Seok; Yoon, Cheolho; Cho, Myung-Haing; Kim, Younghun; Kim, Jae-Ho

    2014-08-17

    In this study, we identified the toxic response of pristine single-walled carbon nanotubes (P-SWCNTs) synthesized by HIPCO method in mice and RAW264.7 cells, a murine peritoneal macrophage cell line. P-SWCNT contained a large amount of Fe ion (36 wt%). In the lungs of mice 24 h after intratracheal administration, P-SWCNTs increased the secretion of IL-6 and MCP-1, and the number of total cells, the portion of neutrophils, lymphocytes, and eosinophils, also significantly increased at a 100 μg/mL of concentration. In RAW264.7 cells, cell viability and ATP production decreased in a dose-dependent manner at 24 h after exposure, whereas the generations of ROS and NO were enhanced at all concentrations together with the activation of the MAP kinase pathway. Moreover, the levels of both apoptosis- and autophagy-related proteins and ER stress-related proteins clearly increased, and the concentrations of Fe, Cu, and Zn ions, but not of Mn ions, increased in a dose-dependent manner. TEM images also revealed that P-SWCNTs induced the formation of autophagosome-like vacuoles, the dilatation of the ER, the generation of mitochondrial flocculent densities, and the separation of organelle by disappearance of the cell membrane. Taken together, we suggest that P-SWCNTs cause acute inflammatory response in the lungs of mice, and induce autophagy accompanied with apoptosis through mitochondrial dysfunction and ER stress in RAW264.7 cells. Furthermore, further study is required to elucidate how the physicochemical properties of SWCNTs determine the cell death pathway and an immune response.

  2. Arabidopsis R-SNARE proteins VAMP721 and VAMP722 are required for cell plate formation.

    Directory of Open Access Journals (Sweden)

    Liang Zhang

    Full Text Available BACKGROUND: Cell plate formation during plant cytokinesis is facilitated by SNARE complex-mediated vesicle fusion at the cell-division plane. However, our knowledge regarding R-SNARE components of membrane fusion machinery for cell plate formation remains quite limited. METHODOLOGY/PRINCIPAL FINDINGS: We report the in vivo function of Arabidopsis VAMP721 and VAMP722, two closely sequence-related R-SNAREs, in cell plate formation. Double homozygous vamp721vamp722 mutant seedlings showed lethal dwarf phenotypes and were characterized by rudimentary roots, cotyledons and hypocotyls. Furthermore, cell wall stubs and incomplete cytokinesis were frequently observed in vamp721vamp722 seedlings. Confocal images revealed that green fluorescent protein-tagged VAMP721 and VAMP722 were preferentially localized to the expanding cell plates in dividing cells. Drug treatments and co-localization analyses demonstrated that punctuate organelles labeled with VAMP721 and VAMP722 represented early endosomes overlapped with VHA-a1-labeled TGN, which were distinct from Golgi stacks and prevacuolar compartments. In addition, protein traffic to the plasma membrane, but not to the vacuole, was severely disrupted in vamp721vamp722 seedlings by subcellular localization of marker proteins. CONCLUSION/SIGNIFICANCE: These observations suggest that VAMP721 and VAMP722 are involved in secretory trafficking to the plasma membrane via TGN/early endosomal compartment, which contributes substantially to cell plate formation during plant cytokinesis.

  3. Autophagy is induced by anti-neutrophil cytoplasmic Abs and promotes neutrophil extracellular traps formation.

    Science.gov (United States)

    Sha, Li-Li; Wang, Huan; Wang, Chen; Peng, Hong-Ying; Chen, Min; Zhao, Ming-Hui

    2016-11-01

    Dysregulated neutrophil extracellular traps (NETs) formation contributes to the pathogenesis of anti-neutrophil cytoplasmic Ab (ANCA)-associated vasculitis (AAV). Increasing evidence indicates that autophagy is involved in the process of NETs formation. In this study, we aimed to investigate whether ANCA could induce autophagy in the process of NETs formation. Autophagy was detected using live cell imaging, microtubule-associated protein light chain 3B (LC3B) accumulation and Western blotting. The results showed that autophagy vacuolization was detected in neutrophils treated with ANCA-positive IgG by live cell imaging. This effect was enhanced by rapamycin, the autophagy inducer, and weakened by 3-methyladenine (3-MA), the autophagy inhibitor. In line with these results, the autophagy marker, LC3B, showed a punctate distribution pattern in the neutrophils stimulated with ANCA-positive IgG. In the presence of rapamycin, LC3B accumulation was further increased; however, this effect was attenuated by 3-MA. Moreover, incubated with ANCA-positive IgG, the NETosis rate significantly increased compared with the unstimulated group. And, the rate significantly increased or decreased in the neutrophils pretreated with rapamycin or 3-MA, respectively, as compared with the cells incubated with ANCA-positive IgG. Overall, this study demonstrates that autophagy is induced by ANCA and promotes ANCA-induced NETs formation.

  4. Simple Ontology Format (SOFT)

    Energy Technology Data Exchange (ETDEWEB)

    2011-10-01

    Simple Ontology Format (SOFT) library and file format specification provides a set of simple tools for developing and maintaining ontologies. The library, implemented as a perl module, supports parsing and verification of the files in SOFt format, operations with ontologies (adding, removing, or filtering of entities), and converting of ontologies into other formats. SOFT allows users to quickly create ontologies using only a basic text editor, verify it, and portray it in a graph layout system using customized styles.

  5. Parasitophorous vacuole membrane of Plasmodium knowlesi

    Energy Technology Data Exchange (ETDEWEB)

    Nillni, E.A.; Wallach, D.F.H.

    1986-05-01

    The authors have evaluated the occurrence of host cell membrane protein and parasite protein in the vacuolar membrane (VM) of isolated parasites. Parasites were labeled by incorporation of (/sup 35/S)methionine and by lactoperoxidase-catalyzed /sup 125/I iodination. Of the two prominent /sup 125/I-labeled components, one, not detected by metabolic labeling corresponded in M/sub r/ to erythrocyte band 3 (90 kDa). Trypsinization of radioiodinated parasites for 5' or 20' yield a 35 kDa fragment, not seen in untreated samples and compatible with the trypsin degradation of band 3 from the cytoplasmic side. Tryptic peptide maps of the 35 kDa revealed a very acidic peptide corresponding to the highly anionic tryptic peptide of band 3 showed by others. The second prominent /sup 125/I-labeled VM protein had an M/sub r/ 74,000 corresponding to a protein metabolically labeled with (/sup 35/S)methionine, suggesting it is inserted into the VM by the parasites. Several less prominent proteins labeling with both (/sup 35/S)methionine and /sup 125/I were also detected (140 kDa, 55 kDa, 45 kDa). A faint /sup 125/I-labeled triple (220-230 kDa) is compatible with a trace amounts of spectrin, usually a prominent component of red cell membrane. The results indicate that host cell band 3 is a prominent component of the VM, but that this membrane also contains several parasite-synthesized proteins.

  6. Star Formation for Predictive Primordial Galaxy Formation

    Science.gov (United States)

    Milosavljević, Miloš; Safranek-Shrader, Chalence

    The elegance of inflationary cosmology and cosmological perturbation theory ends with the formation of the first stars and galaxies, the initial sources of light that launched the phenomenologically rich process of cosmic reionization. Here we review the current understanding of early star formation, emphasizing unsolved problems and technical challenges. We begin with the first generation of stars to form after the Big Bang and trace how they influenced subsequent star formation. The onset of chemical enrichment coincided with a sharp increase in the overall physical complexity of star forming systems. Ab-initio computational treatments are just now entering the domain of the predictive and are establishing contact with local observations of the relics of this ancient epoch.

  7. The Conic Benchmark Format

    DEFF Research Database (Denmark)

    Friberg, Henrik A.

    This document constitutes the technical reference manual of the Conic Benchmark Format with le extension: .cbf or .CBF. It unies linear, second-order cone (also known as conic quadratic) and semidenite optimization with mixed-integer variables. The format has been designed with benchmark libraries...... in mind, and therefore focuses on compact and easily parsable representations. The problem structure is separated from the problem data, and the format moreover facilitate benchmarking of hotstart capability through sequences of changes....

  8. The General Amino Acid Permease FfGap1 of Fusarium fujikuroi Is Sorted to the Vacuole in a Nitrogen-Dependent, but Npr1 Kinase-Independent Manner.

    Directory of Open Access Journals (Sweden)

    Andreas Pfannmüller

    Full Text Available The rice pathogenic fungus Fusarium fujikuroi is well known for the production of a broad spectrum of secondary metabolites (SMs such as gibberellic acids (GAs, mycotoxins and pigments. The biosynthesis of most of these SMs strictly depends on nitrogen availability and of the activity of permeases of nitrogen sources, e.g. the ammonium and amino acid permeases. One of the three ammonium permeases, MepB, was recently shown to act not only as a transporter but also as a nitrogen sensor affecting the production of nitrogen-repressed SMs. Here we describe the identification of a general amino acid permease, FfGap1, among the 99 putative amino acid permeases (AAPs in the genome of F. fujikuroi. FfGap1 is able to fully restore growth of the yeast gap1∆ mutant on several amino acids including citrulline and tryptophane. In S. cerevisiae, Gap1 activity is regulated by shuttling between the plasma membrane (nitrogen limiting conditions and the vacuole (nitrogen sufficiency, which we also show for FfGap1. In yeast, the Npr1 serine/threonine kinase stabilizes the Gap1 position at the plasma membrane. Here, we identified and characterized three NPR1-homologous genes, encoding the putative protein kinases FfNpr1-1, FfNpr1-2 and FfNpr1-3 with significant similarity to yeast Npr1. Complementation of the yeast npr1Δ mutant with each of the three F. fujikuroi NPR1 homologues, resulted in partial restoration of ammonium, arginine and proline uptake by FfNPR1-1 while none of the three kinases affect growth on different nitrogen sources and nitrogen-dependent sorting of FfGap1 in F. fujikuroi. However, exchange of the putative ubiquitin-target lysine 9 (K9A and 15 (K15A residues of FfGap1 resulted in extended localization to the plasma membrane and increased protein stability independently of nitrogen availability. These data suggest a similar regulation of FfGap1 by nitrogen-dependent ubiquitination, but differences regarding the role of Fusarium Npr1

  9. Interaction among Btn1p, Btn2p, and Ist2p reveals potential interplay among the vacuole, amino acid levels, and ion homeostasis in the yeast Saccharomyces cerevisiae.

    Science.gov (United States)

    Kim, Yoojin; Chattopadhyay, Subrata; Locke, Sarahjane; Pearce, David A

    2005-02-01

    Btn2p, a novel cytosolic coiled-coil protein in Saccharomyces cerevisiae, was previously shown to interact with and to be necessary for the correct localization of Rhb1p, a regulator of arginine uptake, and Yif1p, a Golgi protein. We now report the biochemical and physical interactions of Btn2p with Ist2p, a plasma membrane protein that is thought to have a function in salt tolerance. A deletion in Btn2p (btn2Delta strains) results in a failure to correctly localize Ist2p, and strains lacking Btn2p and Ist2p (btn2Delta ist2Delta strains) are unable to grow in the presence of 0.5 or 1.0 M NaCl. Btn2p was originally identified as being up-regulated in a btn1Delta strain, which lacks the vacuolar-lysosomal membrane protein, Btn1p, and serves as a model for Batten disease. This up-regulation of Btn2p was shown to contribute to the maintenance of a stable vacuolar pH in the btn1Delta strain. Btn1p was subsequently shown to be required for the optimal transport of arginine into the vacuole. Interestingly, btn1Delta ist2Delta strains are also unable to grow in the presence of 0.5 or 1.0 M NaCl, and ist2Delta suppresses the vacuolar arginine transport defect in btn1Delta strains. Although further investigation is required, we speculate that altered vacuolar arginine transport in btn1Delta strains represents a mechanism for maintaining or balancing cellular ion homeostasis. Btn2p interacts with at least three proteins that are seemingly involved in different biological functions in different subcellular locations. Due to these multiple interactions, we conclude that Btn2p may play a regulatory role across the cell in response to alterations in the intracellular environment that may be caused by changes in amino acid levels or pH, a disruption in protein trafficking, or imbalances in ion homeostasis resulting from either genetic or environmental manipulation.

  10. Formation of multiple networks

    DEFF Research Database (Denmark)

    Magnani, Matteo; Rossi, Luca

    2013-01-01

    we introduce the first network formation model for multiple networks. Network formation models are among the most popular tools in traditional network studies, because of both their practical and theoretical impact. However, existing models are not sufficient to describe the generation of multiple...

  11. Formative Assessment in Context

    Science.gov (United States)

    Oxenford-O'Brian, Julie

    2013-01-01

    This dissertation responds to critical gaps in current research on formative assessment practice which could limit successful implementation of this practice within the K-12 classroom context. The study applies a socio cultural perspective of learning to interpret a cross-case analysis of formative assessment practice occurring during one…

  12. Galaxy Formation Theory

    CERN Document Server

    Benson, Andrew J

    2010-01-01

    We review the current theory of how galaxies form within the cosmological framework provided by the cold dark matter paradigm for structure formation. Beginning with the pre-galactic evolution of baryonic material we describe the analytical and numerical understanding of how baryons condense into galaxies, what determines the structure of those galaxies and how internal and external processes (including star formation, merging, active galactic nuclei etc.) determine their gross properties and evolution. Throughout, we highlight successes and failings of current galaxy formation theory. We include a review of computational implementations of galaxy formation theory and assess their ability to provide reliable modeling of this complex phenomenon. We finish with a discussion of several "hot topics" in contemporary galaxy formation theory and assess future directions for this field.

  13. Usage Record Format Recommendation

    CERN Document Server

    Nilsen, J.K.; Muller-Pfeerkorn, R

    2013-01-01

    For resources to be shared, sites must be able to exchange basic accounting and usage data in a common format. This document describes a common format which enables the exchange of basic accounting and usage data from different resources. This record format is intended to facilitate the sharing of usage information, particularly in the area of the accounting of jobs, computing, memory, storage and cloud usage but with a structure that allows an easy extension to other resources. This document describes the Usage Record components both in natural language form and annotated XML. This document does not address how these records should be used, nor does it attempt to dictate the format in which the accounting records are stored. Instead, it denes a common exchange format. Furthermore, nothing is said regarding the communication mechanisms employed to exchange the records, i.e. transport layer, framing, authentication, integrity, etc.

  14. When efficient star formation drives cluster formation

    CERN Document Server

    Parmentier, G

    2008-01-01

    We investigate the impact of the star formation efficiency in cluster forming cores on the evolution of the mass in star clusters over the age range 1-100Myr, when star clusters undergo their infant weight-loss/mortality phase. Assuming a constant formation rate of gas-embedded clusters and a weak tidal field, we show that the ratio between the total mass in stars bound to the clusters over that age range and the total mass in stars initially formed in gas-embedded clusters is a strongly increasing function of the averaged local SFE, with little influence from any assumed core mass-radius relation. Our results suggest that, for young starbursts with estimated tidal field strength and known recent star formation history, observed cluster-to-star mass ratios, once corrected for the undetected clusters, constitute promising probes of the local SFE, without the need of resorting to gas mass estimates. Similarly, the mass ratio of stars which remain in bound clusters at the end of the infant mortality/weight-loss ...

  15. Challenges in Planet Formation

    CERN Document Server

    Morbidelli, Alessandro

    2016-01-01

    Over the past two decades, large strides have been made in the field of planet formation. Yet fundamental questions remain. Here we review our state of understanding of five fundamental bottlenecks in planet formation. These are: 1) the structure and evolution of protoplanetary disks; 2) the growth of the first planetesimals; 3) orbital migration driven by interactions between proto-planets and gaseous disk; 4) the origin of the Solar System's orbital architecture; and 5) the relationship between observed super-Earths and our own terrestrial planets. Given our lack of understanding of these issues, even the most successful formation models remain on shaky ground.

  16. Manuel UNIMARC format bibliographique

    CERN Document Server

    2007-01-01

    This manual is the French translation of the second edition of UNIMARC Manual: bibliographic format published in English in 1994 and completed by 5 updates published from 1996 to 2005. This 5th French edition is composite. It reproduces identically a part of the 4th edition published in 2002 and, for the fields of the format modified in the Update 5, it offers a new more structured presentation. This is a handbook dedicated to French-speaking users of the UNIMARC format for bibliographic descriptions.

  17. Methemoglobin formation by paraquat.

    Directory of Open Access Journals (Sweden)

    Watanabe,Shinsaku

    1982-12-01

    Full Text Available Paraquat is a broad spectrum herbicide known to be highly lethal to man and animals. Its toxicity is characterized by acute lung injury. Paraquat produces such toxic effects through the generation of the superoxide anion according to one proposed mechanism. The present experiment, methemoglobin formation was demonstrated after incubation of oxyhemoglobin with paraquat. The generation of the superoxide anion through the interaction of oxyhemoglobin with paraquat was suggested by chemiluminescence of luminol. Superoxide dismutase (SOD and catalare inhibited methemoglobin formation. The generation of the superoxide anion is discussed in regard to methemoglobin formation by paraquat.

  18. Pictorial Formats. Volume 1. Format Development

    Science.gov (United States)

    1982-02-01

    the pilot to help him evaluate when to change course to avoid objects in TF/TA modes. In addition to the projected flight path, there are commanded...TYPESFigMASTER5 Wepo Saus clr ase 00I 16I AI 󈧋TRA- X PI prI MEN p~ll It 62 AIR-TO-GROUND _ - PRGRAM ýRELEASE SELECTIONS _ZY - Figure 58 71T ý Weapon...development should always be done within the context oF a baseline aircraft weapon system. Further studies are needed to simulate and evaluate these formats

  19. Notes on Star Formation

    CERN Document Server

    Krumholz, Mark R

    2015-01-01

    This book provides an introduction to the field of star formation at a level suitable for graduate students or advanced undergraduates in astronomy or physics. The structure of the book is as follows. The first two chapters begin with a discussion of observational techniques, and the basic phenomenology they reveal. The goal is to familiarize students with the basic techniques that will be used throughout, and to provide a common vocabulary for the rest of the book. The next five chapters provide a similar review of the basic physical processes that are important for star formation. Again, the goal is to provide a basis for what follows. The remaining chapters discuss star formation over a variety of scales, starting with the galactic scale and working down to the scales of individual stars and their disks. The book concludes with a brief discussion of the clearing of disks and the transition to planet formation. The book includes five problem sets, complete with solutions.

  20. Collision Induced Galaxy Formation

    CERN Document Server

    Balland, C; Schäffer, R

    1997-01-01

    We present a semi-analytical model in which galaxy collisions and strong tidal interactions, both in the field and during the collapse phase of groups and clusters help determine galaxy morphology. From a semi-analytical analysis based on simulation results of tidal collisions (Aguilar & White 1985), we propose simple rules for energy exchanges during collisions that allow to discriminate between different Hubble types: efficient collisions result in the disruption of disks and substantial star formation, leading to the formation of elliptical galaxies; inefficient collisions allow a large gas reservoir to survive and form disks. Assuming that galaxy formation proceeds in a Omega_0=1 Cold Dark Matter universe, the model both reproduces a number of observations and makes predictions, among which are the redshifts of formation of the different Hubble types in the field. When the model is normalized to the present day abundance of X-ray clusters, the amount of energy exchange needed to produce elliptical gal...

  1. Densities and Kinematic Viscosities for the Systems Benzene + Methyl Formate, Benzene + Ethyl Formate, Benzene + Propyl Formate, and Benzene + Butyl Formate

    DEFF Research Database (Denmark)

    Emmerling, Uwe; Rasmussen, Peter

    1998-01-01

    Densities and kinematic viscosities have been measured for the system benzene + methyl formate at 20°C and for the systems benzene + ethyl formate, benzene + propyl formate, and benzene + butyl formate from 20°C to 50°C. The results for the system benzene + methyl formate have been correlated usi...

  2. Formation peculiarities of tourism documentation

    OpenAIRE

    Zhezhnych, Pavlo; Soprunyuk, Oksana

    2013-01-01

    The article describes formation peculiarities of tourism documentation, the role of tourism data consolidation for unified format creation and the the need to use existing software tools to handle tourism information, formation process of tourism documentation is presented.

  3. Solar System formation

    CERN Document Server

    Crida, A

    2009-01-01

    In this review, three major changes in our understanding of the early history of the Solar System are presented. 1) Early differentiation: A few recent results support the idea that protoplanet formation and differentiation occurred partly simultaneously than CAI formation. First, some iron meteorites, eucrites, and angrites older than the chondrules or even than the CAI have been found. Second, iron meteorites could be debris of early disrupted differentiated planetesimals, scattered from the terrestrial planet region to the Main Belt. Finally, chondrules contain fragments of planetesimal material. 2) Earth and Moon: An equilibration mechanism explains the identical Oxygen isotopic composition of the Earth and the Moon. In addition, it has been shown that the Earth and the Moon mantles have the same 182^W anomaly, in contrast to what was believed before. Consequently, the Moon forming impact should have occurred after the extinction of the 182Hf radioactivity, about 60 Myr after Solar System formation. This ...

  4. Primordial Planet Formation

    CERN Document Server

    Schild, Rudolph E

    2010-01-01

    Recent spacecraft observations exploring solar system properties impact standard paradigms of the formation of stars, planets and comets. We stress the unexpected cloud of microscopic dust resulting from the DEEP IMPACT mission, and the existence of molten nodules in STARDUST samples. And the theory of star formation does not explain the common occurrence of binary and multiple star systems in the standard gas fragmentation scenario. No current theory of planet formation can explain the iron core of the earth, under oceans of water. These difficulties are avoided in a scenario where the planet mass objects form primordially and are today the baryonic dark matter. They have been detected in quasar microlensing and anomalous quasar radio brightening bursts. The primordial planets often concentrate together to form a star, with residual matter seen in pre-stellar accretion discs around the youngest stars. These primordial planet mass bodies were formed of hydrogen-helium, aggregated in dense clumps of a trillion...

  5. Meningococcal biofilm formation

    DEFF Research Database (Denmark)

    Lappann, M.; Haagensen, Janus Anders Juul; Claus, H.

    2006-01-01

    We show that in a standardized in vitro flow system unencapsulated variants of genetically diverse lineages of Neisseria meningitidis formed biofilms, that could be maintained for more than 96 h. Biofilm cells were resistant to penicillin, but not to rifampin or ciprofloxacin. For some strains......, microcolony formation within biofilms was observed. Microcolony formation in strain MC58 depended on a functional copy of the pilE gene encoding the pilus subunit pilin, and was associated with twitching of cells. Nevertheless, unpiliated pilE mutants formed biofilms showing that attachment and accumulation......X alleles was identified among genetically diverse meningococcal strains. PilX alleles differed in their propensity to support autoaggregation of cells in suspension, but not in their ability to support microcolony formation within biofilms in the continuous flow system....

  6. Situated Formative Feedback

    DEFF Research Database (Denmark)

    Lukassen, Niels Bech; Wahl, Christian; Sorensen, Elsebeth Korsgaard

    2016-01-01

    This study addresses the conceptual challenge of providing students with good quality feedback to enhance student learning in an online community of practice (COP). The aim of the study is to identify feedback mechanisms in a virtual learning environment (VLE) and to create a full formative...... feedback episode (FFE) through an online dialogue. The paper argues that dialogue is crucial for student learning and that feedback is not only something the teacher gives to the student. Viewing good quality feedback as social, situated, formative, emphasis is put on the establishment of dialogue. We...... refer to this type of feedback as, Situated Formative Feedback (SFF). As a basis for exploring, identifying and discussing relevant aspects of SFF the paper analyses qualitative data from a Moodle dialogue. Data are embedded in the qualitative analytic program Nvivo and are analysed with a system...

  7. Forces in strategy formation

    DEFF Research Database (Denmark)

    Steensen, Elmer Fly

    2008-01-01

    This chapter proposes that organizational strategy formation should be characterized theoretically as a process that is subject to several interacting forces, rather than represented by separate discrete decisionmodels or theoretic perspectives, as is commonly done in the strategic management...... literature. Based on an extensive review of relevant theory and empirical work in strategic decision-making, organizational change theory, cognitive and social psychology, and strategy processes, seven kinds of ''forces'' - rational, imposed, teleological, learning, political, heuristic, and social...... - are identified as interacting in and having significant influence on the strategy formation process. It is further argued that by applying a holistic ''forces-view'' of the significant and interacting influences on strategy formation, we can better understand the dynamics and challenges in managing the process...

  8. Kuiper Binary Object Formation

    CERN Document Server

    Nazzario, R C; Covington, C; Kagan, D; Hyde, T W

    2005-01-01

    It has been observed that binary Kuiper Belt Objects (KBOs) exist contrary to theoretical expectations. Their creation presents problems to most current models. However, the inclusion of a third body (for example, one of the outer planets) may provide the conditions necessary for the formation of these objects. The presence of a third massive body not only helps to clear the primordial Kuiper Belt but can also result in long lived binary Kuiper belt objects. The gravitational interaction between the KBOs and the third body causes one of four effects; scattering into the Oort cloud, collisions with the growing protoplanets, formation of binary pairs, or creation of a single Kuiper belt object. Additionally, the initial location of the progenitors of the Kuiper belt objects also has a significant effect on binary formation.

  9. Syntactic Formats for Free

    DEFF Research Database (Denmark)

    Klin, Bartek; Sobocinski, Pawel

    2003-01-01

    A framework of Plotkin and Turi’s, originally aimed at providing an abstract notion of bi-simulation, is modified to cover other operational equivalences and preorders. Combined with bi-algebraic methods, it yields a technique for the derivation of syntactic formats for transition system specific......A framework of Plotkin and Turi’s, originally aimed at providing an abstract notion of bi-simulation, is modified to cover other operational equivalences and preorders. Combined with bi-algebraic methods, it yields a technique for the derivation of syntactic formats for transition system...... specifications which guarantee operational preorders to be precongruences. The technique is applied to the trace preorder, the completed trace pre order and the failures preorder. In the latter two cases, new syntactic formats ensuring precongruence properties are introduced....

  10. Sensitivity Data File Formats

    Energy Technology Data Exchange (ETDEWEB)

    Rearden, Bradley T. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)

    2016-04-01

    The format of the TSUNAMI-A sensitivity data file produced by SAMS for cases with deterministic transport solutions is given in Table 6.3.A.1. The occurrence of each entry in the data file is followed by an identification of the data contained on each line of the file and the FORTRAN edit descriptor denoting the format of each line. A brief description of each line is also presented. A sample of the TSUNAMI-A data file for the Flattop-25 sample problem is provided in Figure 6.3.A.1. Here, only two profiles out of the 130 computed are shown.

  11. The formation of stars

    CERN Document Server

    Stahler, Steven W

    2008-01-01

    This book is a comprehensive treatment of star formation, one of the most active fields of modern astronomy. The reader is guided through the subject in a logically compelling manner. Starting from a general description of stars and interstellar clouds, the authors delineate the earliest phases of stellar evolution. They discuss formation activity not only in the Milky Way, but also in other galaxies, both now and in the remote past. Theory and observation are thoroughly integrated, with the aid of numerous figures and images. In summary, this volume is an invaluable resource, both as a text f

  12. Tea aroma formation

    Directory of Open Access Journals (Sweden)

    Chi-Tang Ho

    2015-03-01

    Full Text Available Besides water, tea is one of the most popular beverages around the world. The chemical ingredients and biological activities of tea have been summarized recently. The current review summarizes tea aroma compounds and their formation in green, black, and oolong tea. The flavor of tea can be divided into two categories: taste (non-volatile compounds and aroma (volatile compounds. All of these aroma molecules are generated from carotenoids, lipids, glycosides, etc. precursors, and also from Maillard reaction. In the current review, we focus on the formation mechanism of main aromas during the tea manufacturing process.

  13. Shape formation algorithm

    OpenAIRE

    2016-01-01

    This project concerns the implementation of a decentralized algorithm for shape formation. The first idea was to test this algorithm with a swarm of autonomous drones but, due to the lack of time and the complexity of the project, the work was just developed in 2D and in simulation.

  14. Isothermal Martensite Formation

    DEFF Research Database (Denmark)

    Villa, Matteo

    leading to isothermal kinetics acquired new practical relevance because of the identification of isothermal martensite formation as the most likely process responsible for enhanced performances of sub-zero Celsius treated high carbon steel products. In the present work, different iron based alloys......Isothermal (i.e. time dependent) martensite formation in steel was first observed in the 40ies of the XXth century and is still treated as an anomaly in the description of martensite formation which is considered as a-thermal (i.e. independent of time). Recently, the clarification of the mechanism...... are chosen to investigate time dependent martensite formation. Among them, a Fe-11wt%Ni-0.6wt%C model alloy and Fe-1.6wt%Cr-1wt%C (AISI 52100), Fe-17wt%Cr-7wt%Ni (AISI 631) and Fe-16wt%Cr-5wt%Ni (AISI 630) commercial steels. The investigation was performed with in situ magnetometry, dilatometry, synchrotron...

  15. PAHs and star formation

    NARCIS (Netherlands)

    Tielens, AGGM; Peeters, E; Bakes, ELO; Spoon, HWW; Hony, S; Johnstone, D; Adams, FC; Lin, DNC; Neufeld, DA; Ostriker, EC

    2004-01-01

    Strong IR emission features at 3.3, 6.2, 7.7, 8.6, and 11.2 mum are a common characteristic of regions of massive star formation. These features are carried by large (similar to 50 C-atom) Polycyclic Aromatic Hydrocarbon molecules which are pumped by the strong FUV photon flux from these stars. Thes

  16. BISAC Variable Format.

    Science.gov (United States)

    Information Technology and Libraries, 1983

    1983-01-01

    Presents revision of Book Industry Systems Advisory Committee (BISAC) format designed specifically for electronic transmission of purchase orders for monograph or series titles combining fixed and variable length data fields which was approved in January 1983. Special characters, sample address descriptions, summary of fixed records, glossary, and…

  17. Reference Citation Format

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    <正>The format for citations in text and for bibliographic references follows the Publication Manual of the American Psychological Association(5thEd.,2001)and GB/T 7714-2005.The citation of printed word should be ordered in number as it appears in the text of the submitted

  18. Kepler Planet Formation

    Science.gov (United States)

    Lissauer, Jack J.

    2015-01-01

    Kepler has vastly increased our knowledge of planets and planetary systems located close to stars. The new data shows surprising results for planetary abundances, planetary spacings and the distribution of planets on a mass-radius diagram. The implications of these results for theories of planet formation will be discussed.

  19. Molecular Mechanisms of Autophagy in Plants: Role of ATG8 Proteins in Formation and Functioning of Autophagosomes.

    Science.gov (United States)

    Ryabovol, V V; Minibayeva, F V

    2016-04-01

    Autophagy is an efficient way of degradation and removal of unwanted or damaged intracellular components in plant cells. It plays an important role in recycling of intracellular structures (during starvation, removal of cell components formed during plant development or damaged by various stress factors) and in programmed cell death. Morphologically, autophagy is characterized by the formation of double-membrane vesicles called autophagosomes, which are essential for the isolation and degradation of cytoplasmic components. Among autophagic (ATG) proteins, ATG8 from the ubiquitin-like protein family plays a key role in autophagosome formation. ATG8 is also involved in selective autophagy, fusion of autophagosome with the vacuole, and some other intracellular processes not associated with autophagy. In contrast to yeasts that carry a single ATG8 gene, plants have multigene ATG8 families. The reason for such great ATG8 diversity in plants remains unclear. It is also unknown whether all members of the ATG8 family are involved in the formation and functioning of autophagosomes. To answer these questions, the identification of the structure and the possible functions of plant proteins from ATG8 family is required. In this review, we analyze the structures of ATG8 proteins from plants and their homologs from yeast and animal cells, interactions of ATG8 proteins with functional ligands, and involvement of ATG8 proteins in different metabolic processes in eukaryotes.

  20. Identification of essential histidine residues involved in heme binding and Hemozoin formation in heme detoxification protein from Plasmodium falciparum.

    Science.gov (United States)

    Nakatani, Keisuke; Ishikawa, Haruto; Aono, Shigetoshi; Mizutani, Yasuhisa

    2014-08-20

    Malaria parasites digest hemoglobin within a food vacuole to supply amino acids, releasing the toxic product heme. During the detoxification, toxic free heme is converted into an insoluble crystalline form called hemozoin (Hz). Heme detoxification protein (HDP) in Plasmodium falciparum is one of the most potent of the hemozoin-producing enzymes. However, the reaction mechanisms of HDP are poorly understood. We identified the active site residues in HDP using a combination of Hz formation assay and spectroscopic characterization of mutant proteins. Replacement of the critical histidine residues His122, His172, His175, and His197 resulted in a reduction in the Hz formation activity to approximately 50% of the wild-type protein. Spectroscopic characterization of histidine-substituted mutants revealed that His122 binds heme and that His172 and His175 form a part of another heme-binding site. Our results show that the histidine residues could be present in the individual active sites and could be ligated to each heme. The interaction between heme and the histidine residues would serve as a molecular tether, allowing the proper positioning of two hemes to enable heme dimer formation. The heme dimer would act as a seed for the crystal growth of Hz in P. falciparum.

  1. 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.

  2. Simulating Cosmic Structure Formation

    CERN Document Server

    Weinberg, D H; Hernquist, L E; Weinberg, David H.; Katz, Neal; Hernquist, Lars

    1997-01-01

    We describe cosmological simulation techniques and their application to studies of cosmic structure formation, with particular attention to recent hydrodynamic simulations of structure in the high redshift universe. Collisionless N-body simulations with Gaussian initial conditions produce a pattern of sheets, filaments, tunnels, and voids that resembles the observed large scale galaxy distribution. Simulations that incorporate gas dynamics and dissipation form dense clumps of cold gas with sizes and masses similar to the luminous parts of galaxies. Models based on inflation and cold dark matter predict a healthy population of high redshift galaxies, including systems with star formation rates of 20 M_{\\sun}/year at z=6. At z~3, most of the baryons in these models reside in the low density intergalactic medium, which produces fluctuating Lyman-alpha absorption in the spectra of background quasars. The physical description of this ``Lyman-alpha forest'' is particularly simple if the absorption spectrum is viewe...

  3. Emptiness Formation Probability

    Science.gov (United States)

    Crawford, Nicholas; Ng, Stephen; Starr, Shannon

    2016-08-01

    We present rigorous upper and lower bounds on the emptiness formation probability for the ground state of a spin-1/2 Heisenberg XXZ quantum spin system. For a d-dimensional system we find a rate of decay of the order {exp(-c L^{d+1})} where L is the sidelength of the box in which we ask for the emptiness formation event to occur. In the {d=1} case this confirms previous predictions made in the integrable systems community, though our bounds do not achieve the precision predicted by Bethe ansatz calculations. On the other hand, our bounds in the case {d ≥ 2} are new. The main tools we use are reflection positivity and a rigorous path integral expansion, which is a variation on those previously introduced by Toth, Aizenman-Nachtergaele and Ueltschi.

  4. Cosmic structure formation

    Science.gov (United States)

    Bertschinger, Edumund

    1994-01-01

    This article reviews the prevailing paradigm for how galaxies and larger structures formed in the universe: gravitational instability. Basic observational facts are summarized to motivate the standard cosmological framework underlying most detailed investigations of structure formation. The observed univers approaches spatial uniformity on scales larger than about 10(exp 26) cm. On these scales gravitational dynamics is almost linear and therefore relatively easy to relate to observations of large-scale structure. On smaller scales cosmic structure is complicated not only by nonlinear gravitational clustering but also by nonlinear nongravitational gas dynamical processes. The complexity of these phenomena makes galaxy formation one of the grand challenge problems of the physical sciences. No fully satisfactory theory can presently account in detail for the observed cosmic structure. However, as this article summarizes, significant progress has been made during the last few years.

  5. Un dilemme de formation

    OpenAIRE

    Fields, Marjorie; Marvel, David

    2013-01-01

    Le Master of Art in Teaching prépare à l’enseignement dans l’État de l’Alaska en un an. Les candidats doivent faire preuve d’un bon niveau dans leur discipline pour avoir accès à la formation qui comprend des études théoriques et des stages. Un système d’évaluation de l’enseignant stagiaire et de l’enseignant en fonction permet d’améliorer et de faire évoluer constamment cette formation. Les objectifs de compétence présentés étonneront plus d’un lecteur européen. The Master of Arts in Teac...

  6. The format of things

    DEFF Research Database (Denmark)

    Jørnø, Rasmus Leth

    of much older interfaces that are discussed in philosophy and cognition theory under headlines such as ‘perception,’ ‘cognition’ and ‘representation.’ The conception that is disputed is that the primary way of making sense of the world is to deal with things. In the course of the dissertation...... or surpasses speech and writing. The well from which we draw our design ideas for novel interfaces is therefore needlessly restricted by a format that has outlived its purpose. The objective of the thesis is to dismantle the format of things as well as to sketch out novel paths of inquiry for new interfaces....... The dissertation is based on ideas found in philosophy, Human-Computer Interaction, Cognition theory (enacted, embodied, embedded, extended, situated and distributed), cybernetics, ecological theory, and sociology. The intention is not to take credit for the insight that the world has to be considered in dynamic...

  7. Formate-assisted pyrolysis

    Energy Technology Data Exchange (ETDEWEB)

    DeSisto, William Joseph; Wheeler, Marshall Clayton; van Heiningen, Adriaan R. P.

    2015-03-17

    The present invention provides, among other thing, methods for creating significantly deoxygenated bio-oils form biomass including the steps of providing a feedstock, associating the feedstock with an alkali formate to form a treated feedstock, dewatering the treated feedstock, heating the dewatered treated feedstock to form a vapor product, and condensing the vapor product to form a pyrolysis oil, wherein the pyrolysis oil contains less than 30% oxygen by weight.

  8. Galaxy Formation and Evolution

    Science.gov (United States)

    Nagamine, Kentaro; Reddy, Naveen; Daddi, Emanuele; Sargent, Mark T.

    2016-07-01

    In this chapter, we discuss the current status of observational and computational studies on galaxy formation and evolution. In particular, a joint analysis of star-formation rates (SFRs), stellar masses, and metallicities of galaxies throughout cosmic time can shed light on the processes by which galaxies build up their stellar mass and enrich the environment with heavy elements. Comparison of such observations and the results of numerical simulations can give us insights on the physical importance of various feedback effects by supernovae and active galactic nuclei. In Sect. 1, we first discuss the primary methods used to deduce the SFRs, stellar masses, and (primarily) gas-phase metallicities in high-redshift galaxies. Then, we show how these quantities are related to each other and evolve with time. In Sect. 2, we further examine the distribution of SFRs in galaxies following the `Main Sequence' paradigm. We show how the so-called `starbursts' display higher specific SFRs and SF efficiencies by an order of magnitude. We use this to devise a simple description of the evolution of the star-forming galaxy population since z ˜3 that can successfully reproduce some of the observed statistics in the infrared (IR) wavelength. We also discuss the properties of molecular gas. In Sect. 3, we highlight some of the recent studies of high-redshift galaxy formation using cosmological hydrodynamic simulations. We discuss the physical properties of simulated galaxies such as luminosity function and escape fraction of ionizing photons, which are important statistics for reionization of the Universe. In particular the escape fraction of ionizing photons has large uncertainties, and studying gamma-ray bursts (which is the main topic of this conference) can also set observational constraints on this uncertain physical parameter as well as cosmic star formation rate density.

  9. Reference Citation Format

    Institute of Scientific and Technical Information of China (English)

    2012-01-01

    <正>The format for citations in text and for bibliographic references follows GB/T 7714—2005.The citation should be ordered in number as it appears in the text of the submitted article.For journal article Sun,Y.,Li,B.,&Qu,J.F.Design and implementation of library intelligent IM reference robot.New Technology of Library and Information Service(in Chinese),2011,205:88–92.

  10. Reference Citation Format

    Institute of Scientific and Technical Information of China (English)

    2014-01-01

    <正>The format for citations in text and for bibliographic references follows GB/T 7714—2005.The citation should be ordered in number as it appears in the text of the submitted article.For journal article Sun,Y.,Li,B.,&Qu,J.F.Design and implementation of library intelligent IM reference robot.New Technology of Library and Information Service(in Chinese),

  11. Frost formation with salt

    OpenAIRE

    Guadarrama-Cetina, J.; Mongruel, A; González-Viñas, W; Beysens, D. A.

    2015-01-01

    The formation of frost in presence of salt (NaCl) crystal is experimentally investigated on a hydrophobic surface. It presents several remarkable features due to the interplay of salty-water saturation pressure evolution, initially lower than the saturation pressure of ice and water, and the percolating propagation of ice dendrites from defects throughout the supercooled water droplet pattern. In particular, it is remarkable that nucleation of supercooled water and/or ice is prevented around ...

  12. Reference Citation Format

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    <正>The format for citations in text and for bibliographic references follows the Publication Manual of the American Psychological Association(5thEd.,2001)and GB/T 7714-2005.The citation of printed word should be ordered in number as it appears in the text of the submitted article.For journal article1 Goodrum,A.A.,McCain,K.W.,&Lawrence,S.,et al.Scholarly publishing in the Internet age:

  13. Reference Citation Format

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    <正>The format for citations in text and for bibliographic references follows the Publication Manual of the American Psychological Association(5thEd.,2001)and GB/T 7714-2005.The citation of printed word should be ordered in number as it appears in the text of the submitted article.For journal article1 Goodrum,A.A.,McCain,K.W.,&Lawrence,S.,et al.Scholarly publishing in the Internet

  14. Reference Citation Format

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    <正>The format for citations in text and for bibliographic references follows the Publication Manual of the American Psychological Association(5thEd.,2001)and GB/T 7714-2005.The citation of printed word should be ordered in number as it appears in the text of the submitted article.For journal article1 Goodrum,A.A.,McCain,K.W.,&Lawrence,S.,et al.Scholarly publishing in the Internet

  15. Reference Citation Format

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    <正>The format for citations in text and for bibliographic references follows the Publication Manual of the American Psychological Association(5thEd.,2001)and GB/T 7714-2005.The citation of printed word should be ordered in number as it appears in the text of the submitted article.For journal article1 Goodrum,A.A.,McCain,K.W.,&Lawrence,S.,et al.Scholarly publishing in the Internet age:

  16. Kuiper Binary Object Formation

    OpenAIRE

    Nazzario, R. C.; Orr, K.; Covington, C.; Kagan, D.; Hyde, T. W.

    2005-01-01

    It has been observed that binary Kuiper Belt Objects (KBOs) exist contrary to theoretical expectations. Their creation presents problems to most current models. However, the inclusion of a third body (for example, one of the outer planets) may provide the conditions necessary for the formation of these objects. The presence of a third massive body not only helps to clear the primordial Kuiper Belt but can also result in long lived binary Kuiper belt objects. The gravitational interaction betw...

  17. Tea aroma formation

    OpenAIRE

    Chi-Tang Ho; Xin Zheng; Shiming Li

    2015-01-01

    Besides water, tea is one of the most popular beverages around the world. The chemical ingredients and biological activities of tea have been summarized recently. The current review summarizes tea aroma compounds and their formation in green, black, and oolong tea. The flavor of tea can be divided into two categories: taste (non-volatile compounds) and aroma (volatile compounds). All of these aroma molecules are generated from carotenoids, lipids, glycosides, etc. precursors, and also from Ma...

  18. Formation of Lunar Swirls

    CERN Document Server

    Bamford, R A; Cruz, F; Kellett, B J; Fonseca, R A; Silva, L O; Trines, R M G M; Halekas, J S; Kramer, G; Harnett, E; Cairns, R A; Bingham, R

    2015-01-01

    In this paper we show a plausible mechanism that could lead to the formation of the Dark Lanes in Lunar Swirls, and the electromagnetic shielding of the lunar surface that results in the preservation of the white colour of the lunar regolith. We present the results of a fully self-consistent 2 and 3 dimensional particle-in-cell simulations of mini-magnetospheres that form above the lunar surface and show that they are consistent with the formation of `lunar swirls' such as the archetypal formation Reiner Gamma. The simulations show how the microphysics of the deflection/shielding of plasma operates from a kinetic-scale cavity, and show that this interaction leads to a footprint with sharp features that could be the mechanism behind the generation of `dark lanes'. The physics of mini-magnetospheres is described and shown to be controlled by space-charge fields arising due to the magnetized electrons and unmagnetized ions. A comparison between model and observation is shown for a number of key plasma parameters...

  19. The Star Formation Camera

    CERN Document Server

    Scowen, Paul A; Beasley, Matthew; Calzetti, Daniela; Desch, Steven; Fullerton, Alex; Gallagher, John; Lisman, Doug; Macenka, Steve; Malhotra, Sangeeta; McCaughrean, Mark; Nikzad, Shouleh; O'Connell, Robert; Oey, Sally; Padgett, Deborah; Rhoads, James; Roberge, Aki; Siegmund, Oswald; Shaklan, Stuart; Smith, Nathan; Stern, Daniel; Tumlinson, Jason; Windhorst, Rogier; Woodruff, Robert

    2009-01-01

    The Star Formation Camera (SFC) is a wide-field (~15'x19, >280 arcmin^2), high-resolution (18x18 mas pixels) UV/optical dichroic camera designed for the Theia 4-m space-borne space telescope concept. SFC will deliver diffraction-limited images at lambda > 300 nm in both a blue (190-517nm) and a red (517-1075nm) channel simultaneously. Our aim is to conduct a comprehensive and systematic study of the astrophysical processes and environments relevant for the births and life cycles of stars and their planetary systems, and to investigate and understand the range of environments, feedback mechanisms, and other factors that most affect the outcome of the star and planet formation process. This program addresses the origins and evolution of stars, galaxies, and cosmic structure and has direct relevance for the formation and survival of planetary systems like our Solar System and planets like Earth. We present the design and performance specifications resulting from the implementation study of the camera, conducted ...

  20. 荧光蛋白在植物活细胞液泡成像中的应用研究进展%Recent Progress in Living Cell Imaging of Plant Vacuole Using Fluorescent-protein Transgenic Lines and Three-dimensional Imaging

    Institute of Scientific and Technical Information of China (English)

    叶庆亮; 曹建华

    2011-01-01

    高等植物细胞中,液泡在各种细胞信号转导事件和形态建成中起着重要的作用.这一细胞内部结构在分裂和分化期间其功能和形状不断地变化着.为分析这些连续的变化,绿色荧光蛋白(green fluorescent protein,GFP)及其他荧光蛋白活体标记技术普遍用来跟踪特异蛋白的定位及变动.为使液泡可视成像,有几种途径可用来选择适当的荧光蛋白融合伴侣,如液泡膜内在蛋白和构造蛋白相关蛋白等就非常适合应用于液泡成像.此外,三维重建法在定位细胞内广为分布的细胞器时也不可或缺.同时,等值面表面模化过程对液泡膜成像也非常有用.本文综述液泡的结构、种类和功能,并概括各种融合的绿色荧光蛋白在植物活细胞液泡成像中的应用及其三维成像技术的研究进展.%In high plant cells, vacuoles play important roles in a variety of cellular events, including cell division,morphogenesis, and signal transduction.These intracellular structures undergo dynamic changes in their shapes and functions during cell division and differentiation, and to analyze these sequential structural changes, the vital labeling technique, using the green-fluorescent protein or other fluorescent proteins, has commonly been used to follow the localization and translocation of specific proteins.To visualize vacuoles, the tonoplast-intrinsic proteins and syntaxin-related proteins are available for selecting the appropriate fluorescent-protein fusion partner for vacuolar imaging.In addition, three-dimensional reconstruction methods are indispensable for localizing the widely distributed organelles within the cell.The maximum intensity projection method is suitable for cytoskeletal structures, while contour-based surface modeling possesses many advantages for vacuolar membranes.In this article, we summarize the plant vacuoles and the recent progress in living cell imaging of the plant vacuoles using various fusions

  1. The formation of life

    CERN Document Server

    Kurucz, R L

    2000-01-01

    The formation of life is an automatic stage in the consolidation of rocky or "terrestrial" planets. The organic (=carbonaceous) matter, light elements, gases, and water must "float" toward the surface and the heavier metals must sink toward the center. Random processes in the molecular soup that fills microfractures in unmelted crust eventually produce self-replicating microtubules. In an appendix I suggest that some primordial crust remains because there is not enough consolidation energy to melt the whole planet. Energy is lost when iron planetesimals first partially melt and then coalesce to form the molten iron planetary core. Stony planetesimals accrete onto the surface of an already consolidated core.

  2. Formation control of AAUSHIP

    DEFF Research Database (Denmark)

    Østergaard, Nick; Dam, Jeppe; Larsen, Jesper Abildgaard

    2015-01-01

    Many maritime mapping tasks are today carried out by large research ships, which are very costly to operate. As a way to overcome this, a number of small surveying vessels have been developed called AAUSHIP. In order to efficiently map the an area with such smaller vessels, it is important that s...... that several vessels are able to corporate on the task at hand. In this paper, the developed formation control strategy for the AAUSHIP series of vessels is presented, along with simulation results, which confirms, that the algorithm works as intended....

  3. Bouyei word formation

    Directory of Open Access Journals (Sweden)

    Attasith Boonsawasd

    2012-12-01

    Full Text Available The Bouyei language is divided into three vernaculars, the southern vernacular, the central vernacular and the southwestern vernacular. This paper aims to describe the lexicology of the southern vernacular of the Bouyei language focusing on word formation process. Bouyei words are formed by affixing, compounding and reduplicating. First, the affixation consists of prefixing and suffixing. Infixing is not found in this language. Second, the compound is divided into the semantic and syntactic compound. Finally, the reduplication is divided into the simple and complex reduplication. The simple reduplication is normally used to emphasize the meaning of the root or to indicate plurality.

  4. Reference Citation Format

    Institute of Scientific and Technical Information of China (English)

    2011-01-01

    <正>The format for citations in text and for bibliographic references follows the Publication Manual of the American Psychological Association(5thEd.,2001)and GB/T 7714-2005.The citation of printed word should be ordered in number as it appears in the text of the submitted article.For journal article1 Goodrum,A.A.,McCain,K.W.,&Lawrence,S.,et al.Scholarly publishing in the Internet age:A citation analysis of computer science literature.Information Processing and Manage-

  5. Reference Citation Format

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    <正>The format for citations in text and for bibliographic references follows the Publication Manual of the American Psychological Association(5thEd.,2001)and GB/T 7714-2005.The citation of printed word should be ordered in number as it appears in the text of the submitted article.For journal article1 Goodrum,A.A.,McCain,K.W.,&Lawrence,S.,et al.Scholarly publishing in the Internet age:A citation analysis of computer science literature.Information Processing and Management,2001,37:661-675.2 Fernandez,M.,Kadiyska,Y.,&Suciu,D.,et al.SilkRoute:A framework for publishing

  6. Reference Citation Format

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    <正>The format for citations in text and for bibliographic references follows the Publication Manual of the American Psychological Association(5th Ed.,2001)and GB/T 7714-2005.The citation of printed word should be ordered in number as it appears in the text of the submitted article.For journal article1 Goodrum,A.A.,McCain,K.W.,&Lawrence,S.,et al.Scholarly publishing in the Internet age:A citation analysis of computer science literature.Information Processing and Management,2001,37:661-675.

  7. Reference Citation Format

    Institute of Scientific and Technical Information of China (English)

    2011-01-01

    <正>The format for citations in text and for bibliographic references follows the Publication Manual of the American Psychological Association(5thEd.,2001)and GB/T 7714-2005.The citation of printed word should be ordered in number as it appears in the text of the submitted article.For journal article1 Goodrum,A.A.,McCain,K.W.,&Lawrence,S.,et al.Scholarly publishing in the Internet age:A citation analysis of computer science literature.Information Processing and Management,2001,37:661-675.2 Fernandez.M

  8. Star Formation in Irregular Galaxies.

    Science.gov (United States)

    Hunter, Deidre; Wolff, Sidney

    1985-01-01

    Examines mechanisms of how stars are formed in irregular galaxies. Formation in giant irregular galaxies, formation in dwarf irregular galaxies, and comparisons with larger star-forming regions found in spiral galaxies are considered separately. (JN)

  9. Explosions during galaxy formation

    Directory of Open Access Journals (Sweden)

    Hugo Martel

    2001-01-01

    Full Text Available As an idealized model of the e ects of energy release by supernovae during galaxy formation, we consider an explosion at the center of a halo which forms at the intersection of laments in the plane of a cosmological pancake by gravitational instability during pancake collapse. Such halos resemble the virialized objects found in N{body simulations in a CDM universe and, therefore, serve as a convenient, scale{free test{bed model for galaxy formation. ASPH=P3M simulations reveal that such explosions are anisotropic. The energy and metals are channeled into the low density regions, away from the pancake plane. The pancake remains essentially undisturbed, even if the explosion is strong enough to blow away all the gas lo- cated inside the halo at the onset of the explosion and reheat the IGM surrounding the pancake. Infall quickly replenishes this ejected gas and gradually restores the gas fraction as the halo mass continues to grow. Estimates of the collapse epoch and SN energy{release for galaxies of di erent mass in the CDM model can re- late these results to scale{dependent questions of blow{out and blow{away and their implication for early IGM heating and metal enrichment and the creation of dark{matter{dominated dwarf galaxies.

  10. Formation of water bells

    Science.gov (United States)

    Ybert, Christophe; Clanet, Christophe; Bocquet, Lyderic; Duez, Cyril

    2007-11-01

    We study experimentally the situation that consist in a liquid jet impacting normally onto a fixed solid disk. Depending on the experimental conditions, the thin liquid film that spreads onto the solid surface can either pour along the surface, or detach form the disk and form a so-called water bell. The dynamics and the stability of such bells as a function of the hydrodynamic parameters such as the jet and disk diameters or the jet velocity, have already been the object of detailed characterization [1]. This experiment of bell formation appears as the symmetric situation compared to that of a solid body impacting a quiescent liquid. In the latter case, it was recently shown [2] that despite large Re and We numbers, the solid surface characteristics were dramatically influencing the impact scenario. In the present study, we consequently revisit this problem of water bell formation by systematically varying the solid surface characteristics (roughness, surface properties, etc.). It is shown here again that surface parameters strongly influence the domain of bell existence. Our measurements are rationalized by a subtle balance between inertia versus capillary forces and wetting contributions on the liquid film in the ejection region. [1] C. Clanet, J. Fluid Mech., 430, 111-147 (2001) [2] C. Duez et al., Nature Physics, 3, 180-183 (2007)

  11. Parametrising Star Formation Histories

    CERN Document Server

    Simha, Vimal; Conroy, Charlie; Dave, Romeel; Fardal, Mark; Katz, Neal; Oppenheimer, Benjamin D

    2014-01-01

    We examine the star formation histories (SFHs) of galaxies in smoothed particle hydrodynamics (SPH) simulations, compare them to parametric models that are commonly used in fitting observed galaxy spectral energy distributions, and examine the efficacy of these parametric models as practical tools for recovering the physical parameters of galaxies. The commonly used tau-model, with SFR ~ exp(-t/tau), provides a poor match to the SFH of our SPH galaxies, with a mismatch between early and late star formation that leads to systematic errors in predicting colours and stellar mass-to-light ratios. A one-parameter lin-exp model, with SFR ~ t*exp(-t/tau), is much more successful on average, but it fails to match the late-time behavior of the bluest, most actively star-forming galaxies and the passive, "red and dead" galaxies. We introduce a 4-parameter model, which transitions from lin-exp to a linear ramp after a transition time, which describes our simulated galaxies very well. We test the ability of these paramet...

  12. On Formations with Shemetkov Condition

    Institute of Scientific and Technical Information of China (English)

    Wenbin Guo; Lujin Zhu

    2002-01-01

    A subgroup-closed formation F is called a formation with Shemetkov condition if every minimal non-F-group is either a group of prime order or a Schmidt group. In this paper, we give a necessary and sufficient condition for formations with Shemetkov condition. From this, some known important results follow.

  13. Designing for informed group formation

    DEFF Research Database (Denmark)

    Nicolajsen, Hanne Westh; Juel Jacobsen, Alice; Riis, Marianne

    2012-01-01

    A new design ―project preparation‖ preparing for the group formation in problem based project work is proposed and investigated. The main problem is to overcome group formation based on existing relations. The hypothesis is that theme development and group formation are somewhat counterproductive...

  14. Synapse formation and remodeling

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    Synapses are specialized structures that mediate information flow between neurons and target cells,and thus are the basis for neuronal system to execute various functions,including learning and memory.There are around 1011 neurons in the human brain,with each neuron receiving thousands of synaptic inputs,either excitatory or inhibitory.A synapse is an asymmetric structure that is composed of pre-synaptic axon terminals,synaptic cleft,and postsynaptic compartments.Synapse formation involves a number of cell adhesion molecules,extracellular factors,and intracellular signaling or structural proteins.After the establishment of synaptic connections,synapses undergo structural or functional changes,known as synaptic plasticity which is believed to be regulated by neuronal activity and a variety of secreted factors.This review summarizes recent progress in the field of synapse development,with particular emphasis on the work carried out in China during the past 10 years(1999-2009).

  15. Nuclear Pasta Formation

    CERN Document Server

    Schneider, A S; Hughto, J; Berry, D K

    2013-01-01

    The formation of complex nonuniform phases of nuclear matter, known as nuclear pasta, is studied with molecular dynamics simulations containing 51200 nucleons. A phenomenological nuclear interaction is used that reproduces the saturation binding energy and density of nuclear matter. Systems are prepared at an initial density of 0.10fm$^{-3}$ and then the density is decreased by expanding the simulation volume at different rates to densities of 0.01 fm$^{-3}$ or less. An originally uniform system of nuclear matter is observed to form spherical bubbles ("swiss cheese"), hollow tubes, flat plates ("lasagna"), thin rods ("spaghetti") and, finally, nearly spherical nuclei with decreasing density. We explicitly observe nucleation mechanisms, with decreasing density, for these different pasta phase transitions. Topological quantities known as Minkowski functionals are obtained to characterize the pasta shapes. Different pasta shapes are observed depending on the expansion rate. This indicates non equilibrium effects...

  16. Reference Citation Format

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    <正>The format for citations in text and for bibliographic references follows the Publication Manual of the American Psychological Association(5thEd.,2001)and GB/T 7714-2005.The citation of printed word should be ordered in number as it appears in the text of the submitted article.For journal article1 Goodrum,A.A.,McCain,K.W.,&Lawrence,S.,et al.Scholarly publishing in the Internet age:A citation analysis of computer science literature.Information Processing and Management,2001,37:661-675.2 Fernandez,M.,Kadiyska,Y.,&Suciu,D.,et al.SilkRoute:A framework for publishing relational data in XML.ACM Transactions on Database Systems,2002,27(4):438-493.

  17. Reference Citation Format

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    <正>The format for citations in text and for bibliographic references follows the Publication Manual of the American Psychological Association(5thEd.,2001)and GB/T 7714-2005.The citation of printed word should be ordered in number as it appears in the text of the submitted article.For journal article1 Goodrum,A.A.,McCain,K.W.,&Lawrence,S.,et al.Scholarly publishing in the Internet age:A citation analysis of computer science literature.Information Processing and Management,2001,37:661-675.2 Fernandez,M.,Kadiyska,Y.,&Suciu,D.,et al.SilkRoute:A framework for publishing relational data in XML.ACM Transactions on Database Systems,2002,27(4):438-493.

  18. Reference Citation Format

    Institute of Scientific and Technical Information of China (English)

    2011-01-01

    <正>The format for citations in text and for bibliographic references follows the Publication Manual of the American Psychological Association(5thEd.,2001)and GB/T 7714-2005.The citation of printed word should be ordered in number as it appears in the text of the submitted article.For journal article1 Goodrum,A.A.,McCain,K.W.,&Lawrence,S.,et al.Scholarly publishing in the Internet age:A citation analysis of computer science literature.Information Processing and Management,2001,37:661-675.2 Fernandez,M.,Kadiyska,Y.,&Suciu,D.,et al.SilkRoute:A framework for publishing relational data in XML.ACM Transactions on Database Systems,2002,27(4):438-493.

  19. Formation of Service Ecosystems

    DEFF Research Database (Denmark)

    Jonas, Julia M.; Sörhammar, David; Satzger, Gerhard

    – i.e. the “birth phase” (Moore, 2009) of a service ecosystem. This paper, therefore, aims to explore how the somewhat “magic” processes of service ecosystem formation that are being taken for granted actually occur. Methodology/Approach: Building on a review of core elements in the definitions......Purpose: Researchers in several different academic disciplines (such as marketing, information systems, and organization) have focused on investigating service and business ecosystems (e.g. Lusch and Nambisan, 2015; Gawer and Cusumano, 2014; Kude et al. 2012). We reviewed 69 papers in service...... science, operations management, marketing, and organization journals. The majority of these papers focus on describing established service ecosystems,often on a more abstract “meso-level” (Akaka et al., 2015). Key notions are “…self-contained, self-adjusting system[s] of resource integrating actors...

  20. Symbol Formation Reconsidered

    DEFF Research Database (Denmark)

    Wagoner, Brady

    2013-01-01

    them vis-à-vis other research at Clark and in American psychology more generally. The second two articles analyse Werner and Kaplan’s notions of ‘distancing’ and ‘physiognomic metaphor’, showing their roots in naturphilosophie and comparing them with contemporary theories. The last four articles apply......Werner and Kaplan’s Symbol formation was published 50 years ago but its insights have yet to be adequately explored by psychology and other social sciences. This special issue aims to revisit this seminal work in search of concepts to work on key issues facing us today. This introductory article...... begins with a brief outline and contextualization of the book as well as of the articles that this special issue comprises. The first two articles were written by contributors who were part of the Werner era at Clark University. They explore the key concepts of the organismic and development, and situate...

  1. Cosmological Structure Formation

    CERN Document Server

    Primack, Joel R

    2015-01-01

    LCDM is remarkably successful in predicting the cosmic microwave background and large-scale structure, and LCDM parameters have been determined with only mild tensions between different types of observations. Hydrodynamical simulations starting from cosmological initial conditions are increasingly able to capture the complex interactions between dark matter and baryonic matter in galaxy formation. Simulations with relatively low resolution now succeed in describing the overall galaxy population. For example, the EAGLE simulation in volumes up to 100 cubic Mpc reproduces the observed local galaxy mass function nearly as well as semi-analytic models. It once seemed that galaxies are pretty smooth, that they generally grow in size as they evolve, and that they are a combination of disks and spheroids. But recent HST observations combined with high-resolution hydrodynamic simulations are showing that most star-forming galaxies are very clumpy; that galaxies often undergo compaction which reduces their radius and ...

  2. Chinese Affixes and Word Formation

    Directory of Open Access Journals (Sweden)

    Fu Ruomei

    2014-05-01

    Full Text Available Chinese language is one of the typical isolated languages. It lacks morphological variation; part of speech has no morphological signs; the additional component of word formation is less; and the roots never change their forms. The major method of Chinese word formation is the combination of roots according to certain grammatical relations. Although the affix word formation is not part of mainstream Chinese word formation, affix-formation is still an integral part of the Chinese word-formation. Article used literature review, summarized the types and meanings of Chinese affixes. And meanwhile, article analyzed word formation function of Chinese Affixes and quasi-affixes. The Chinese quasi-affixes have stronger capabilities in forming new words, but development direction of Chinese quasi-affixes has to stand the test of time.

  3. 外周血淋巴细胞异常空泡检查对溶酶体贮积病筛查和诊断的价值%Detection of vacuolated peripheral blood lymphocytes in screening for and diagnosis of lysosomal storage diseases

    Institute of Scientific and Technical Information of China (English)

    常杏芝; 刘洁玉; 吴晔; 姜玉武; 熊晖; 王爽; 秦炯

    2011-01-01

    Objective Lysosomal storage diseases are a group of inherited disorders caused by deficiency of lysosomal enzymes or structural components. The manifestations of lysosomal storage diseases are complicated due to different enzyme deficiency. It has been reported that a range of metabolic diseases resulting in abnormal accumulation of metabolic byproducts may exhibit abnormal cytoplasmic vacuolation of lymphocytes. The aim of this study was to elicit the usefulness of vacuolated peripheral lymphocytes detection in screening and diagnosis of lysosomal storage diseases. Method Clinical data of 42 patients who underwent microscopic and electron microscopic examination of peripheral blood specimens in our department were retrospectively evaluated between January 2008 and December 2009. Result Forty-two patients with the suspected lysosomal storage diseases were included, these patients presented with motor and developmental retardation and/or regression. Seizure occurred in 32 patients. Hepatosplenomegaly were found in 4 patients. Three patients presented with declined visual acuity. Atrophy and/or abnormal signals were detected on cranial CT/MRI images in 24 patients. Blood biochemical tests were normal. Serum levels of ammonia, lactic acid and pyruvate were normal. Serum amino acid profiles and urinary organic acid profiles were normal. Serum fatty acid profiles were normal. Vacuolated lymphocytes were detected on microscopic examination of blood film in 14 patients, and 8 of these patients were confirmed to have lysosomal storage disease. Curvilinear body was found on electronic microscopic examination of peripheral lymphocytes specimens in 4 patients, confirming the diagnosis of neuronal ceroid lipofuscinosis. In 3 of these 4 patients, curvilinear body were also found on electronic microscopic examination of skin and/or muscle specimens. Enzyme analysis confirmed the diagnosis of metachromatic leukodystrophy in one patient and Pompe's disease in another patient

  4. Modeling Formation of Globular Clusters: Beacons of Galactic Star Formation

    CERN Document Server

    Gnedin, Oleg Y

    2010-01-01

    Modern hydrodynamic simulations of galaxy formation are able to predict accurately the rates and locations of the assembly of giant molecular clouds in early galaxies. These clouds could host star clusters with the masses and sizes of real globular clusters. I describe current state-of-the-art simulations aimed at understanding the origin of the cluster mass function and metallicity distribution. Metallicity bimodality of globular cluster systems appears to be a natural outcome of hierarchical formation and gradually declining fraction of cold gas in galaxies. Globular cluster formation was most prominent at redshifts z>3, when massive star clusters may have contributed as much as 20% of all galactic star formation.

  5. Planet Formation with Migration

    CERN Document Server

    Chambers, J E

    2006-01-01

    In the core-accretion model, gas-giant planets form solid cores which then accrete gaseous envelopes. Tidal interactions with disk gas cause a core to undergo inward type-I migration in 10^4 to 10^5 years. Cores must form faster than this to survive. Giant planets clear a gap in the disk and undergo inward type-II migration in <10^6 years if observed disk accretion rates apply to the disk as a whole. Type-II migration times exceed typical disk lifetimes if viscous accretion occurs mainly in the surface layers of disks. Low turbulent viscosities near the midplane may allow planetesimals to form by coagulation of dust grains. The radius r of such planetesimals is unknown. If r<0.5 km, the core formation time is shorter than the type-I migration timescale and cores will survive. Migration is substantial in most cases, leading to a wide range of planetary orbits, consistent with the observed variety of extrasolar systems. When r is of order 100m and midplane alpha is of order 3 times 10^-5, giant planets si...

  6. ISLET FORMATION AND REGENERATION

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Objective To explore the mechanisms of differentiation and development of pancreatic endocrine cells as well as pancreatic regeneration. Methods Human embryonic pancreatic tissue at 7-14 weeks of gestation was collected. Diabetes mellitus rat model was induced with 65 mg/kg of streptozotocin. Insulin, glucagon, somatostatin, nestin, and cytokeratin 19 (CK19)of pancreatic tissues were observed by immunohistochemistry. Results At 9 weeks of gestation, pancreatic epithelial cells began to co-express insulin, glucagon, somatostatin, and CK19 before migration. Islet cells gradually congregated along with the increase of aging, and at 14 weeks of gestation histological examination showed islet formation. At 12 weeks of gestation, nestin-positive cells could be seen in the pancreatic mesenchyme. During early embryogenesis, islet cells of pancreatic ducts co-expressed insulin, glucagon, and somatostatin. During pancreatic regeneration after damage, nestin expression of islet cells increased. Conclusion In the early stage of embryogenesis, islet cells of primary pancreatic ducts can be differentiated to multipotential endocrine cells before migration. During tissue regeneration, pancreatic stem cells may differentiate and proliferate to form pancreatic islet.

  7. Formation time of hadronic resonances

    Directory of Open Access Journals (Sweden)

    Vitev Ivan

    2012-11-01

    Full Text Available In heavy-ion collisions, formation time of hadrons of high transverse momentum can play a pivotal role in determining the perturbative dynamics of the final-state parton and particle system. We present methods to evaluate the formation times of light hadrons, hadronic resonances, open heavy flavor and quarkonia. Experimental implications of the short formation times of heavy particles are discussed in light of recent RHIC and LHC data.

  8. Spray formation: an inverse cascade

    CERN Document Server

    Ling, Yue; Tryggvason, Gretar; zaleski, Stephane

    2015-01-01

    We present a study of droplet formation in a gas-liquid mixing layer using direct numerical simulation. It is seen that two mechanisms compete to generate the droplets: fingering at the tip of the waves and hole formation in the thin liquid sheet. The three dimensional liquid structures are much shorter than the longitudinal wavelength of the instability at the first instant of their formation. As time evolves, the structures evolves to larger and larger scales, in a way similar to the inverse cascade of length scales in droplet impact and impact crown formation.

  9. Glass formation - A contemporary view

    Science.gov (United States)

    Uhlmann, D. R.

    1983-01-01

    The process of glass formation is discussed from several perspectives. Particular attention is directed to kinetic treatments of glass formation and to the question of how fast a given liquid must be cooled in order to form a glass. Specific consideration is paid to the calculation of critical cooling rates for glass formation, to the effects of nucleating heterogeneities and transients in nucleation on the critical cooling rates, to crystallization on reheating a glass, to the experimental determination of nucleation rates and barriers to crystal nucleation, and to the characteristics of materials which are most conducive to glass formation.

  10. Formative Assessment Probes: Is It a Rock? Continuous Formative Assessment

    Science.gov (United States)

    Keeley, Page

    2013-01-01

    A lesson plan is provided for a formative assessment probe entitled "Is It a Rock?" This probe is designed for teaching elementary school students about rocks through the use of a formative assessment classroom technique (FACT) known as the group Frayer Model. FACT activates students' thinking about a concept and can be used to…

  11. Satellite formation flying relative dynamics, formation design, fuel optimal maneuvers and formation maintenance

    CERN Document Server

    Wang, Danwei; Poh, Eng Kee

    2017-01-01

    This book systematically describes the concepts and principles for multi-satellite relative motion, passive and near passive formation designs, trajectory planning and control for fuel optimal formation maneuvers, and formation flying maintenance control design. As such, it provides a sound foundation for researchers and engineers in this field to develop further theories and pursue their implementations. Though satellite formation flying is widely considered to be a major advance in space technology, there are few systematic treatments of the topic in the literature. Addressing that gap, the book offers a valuable resource for academics, researchers, postgraduate students and practitioners in the field of satellite science and engineering.

  12. The Principal as Formative Coach

    Science.gov (United States)

    Nidus, Gabrielle; Sadder, Maya

    2011-01-01

    Formative coaching, an approach that uses student work as the foundation for mentoring and professional development, can help principals become more effective instructional leaders. In formative coaching, teaches and coaches analyze student work to determine next steps for instruction. This article shows how a principal can use the steps of the…

  13. Sparse Matrix Vector Processing Formats

    NARCIS (Netherlands)

    Stathis, P.T.

    2004-01-01

    In this dissertation we have identified vector processing shortcomings related to the efficient storing and processing of sparse matrices. To alleviate existent problems we propose two storage formats denoted as Block Based Compression Storage (BBCS) format and Hierarchical Sparse Matrix (HiSM) stor

  14. Tariffs Formation on oil transportation

    Science.gov (United States)

    Glyzina, T. S.; Kolbysheva, Yu. V.; Grivtsova, I. S.; Dmitrieva, N. V.

    2016-09-01

    Oil transportation via trunk pipelines is an important part of the oil industry's activity. The main instrument of tariff regulation is the method of tariffs formation. Three methods of tariffs formation such as the method of economically justified costs (the Cost plus method), the method of economically justified return on investment capital (the RAB method), and the method of tariffs indexation were considered.

  15. Identity formation in multiparty negotiations

    NARCIS (Netherlands)

    Swaab, R; Postmes, T.; Spears, R.

    2008-01-01

    Based on the recently proposed Interactive Model of Identity Formation, we examine how top-down deductive and bottom-up inductive identity formations influence intentions and behaviour in multiparty negotiations. Results show that a shared identity can be deduced from the social context through reco

  16. Chain formation of metal atoms

    DEFF Research Database (Denmark)

    Bahn, Sune Rastad; Jacobsen, Karsten Wedel

    2001-01-01

    The possibility of formation of single-atomic chains by manipulation of nanocontacts is studied for a selection of metals (Ni, Pd, Pt, Cu, Ag, Au). Molecular dynamics simulations show that the tendency for chain formation is strongest for Au and Pt. Density functional theory calculations indicate...

  17. Bundle Formation in Biomimetic Hydrogels

    NARCIS (Netherlands)

    Jaspers, Maarten; Pape, A C H; Voets, Ilja K; Rowan, Alan E; Portale, Giuseppe; Kouwer, Paul H J

    2016-01-01

    Bundling of single polymer chains is a crucial process in the formation of biopolymer network gels that make up the extracellular matrix and the cytoskeleton. This bundled architecture leads to gels with distinctive properties, including a large-pore-size gel formation at very low concentrations and

  18. SAS FORMATS: USES AND ABUSES

    Science.gov (United States)

    SAS formats are a very powerful tool. They allow you to display the data in a more readable manner without modifying it. Formats can also be used to group data into categories for use in various procedures like PROC FREQ, PROC TTEST, and PROC MEANS (as a class variable). As ...

  19. Formation depths of Fraunhofer lines

    CERN Document Server

    Gurtovenko, E A

    2015-01-01

    We have summed up our investigations performed in 1970--1993. The main task of this paper is clearly to show processes of formation of spectral lines as well as their distinction by validity and by location. For 503 photospheric lines of various chemical elements in the wavelength range 300--1000 nm we list in Table the average formation depths of the line depression and the line emission for the line centre and on the half-width of the line, the average formation depths of the continuum emission as well as the effective widths of the layer of the line depression formation. Dependence of average depths of line depression formation on excitation potential, equivalent widths, and central line depth are demonstrated by iron lines.

  20. Formative assessments as pedagogic tools

    Directory of Open Access Journals (Sweden)

    Ashwin Kumar

    2013-03-01

    Full Text Available This article aims at presenting a state of the art status of formative assessment as a pedagogic tool. To this end, a brief developmental account of different modes of assessment over the last decades will be presented first. Then, formative assessment will be discussed in its constructivist guise. The present literature on assessment suggests that assessment for learning (formative assessment not only represents an assessment tool but it also serves as a pedagogic tool to enhance learning and thinking. It has also gone to lengths to affect the design of classroom tasks and activities. Attempts have been made to delineate the underlying principles of formative assessment which can be used to picture the formation process of learners’ knowledge and development. Subsequently, alternative assessment techniques of which the present article will give an account have been suggested by scholars to operationalize these principles.

  1. Physics of primordial star formation

    Science.gov (United States)

    Yoshida, Naoki

    2012-09-01

    The study of primordial star formation has a history of nearly sixty years. It is generally thought that primordial stars are one of the key elements in a broad range of topics in astronomy and cosmology, from Galactic chemical evolution to the formation of super-massive blackholes. We review recent progress in the theory of primordial star formation. The standard theory of cosmic structure formation posits that the present-day rich structure of the Universe developed through gravitational amplification of tiny matter density fluctuations left over from the Big Bang. It has become possible to study primordial star formation rigorously within the framework of the standard cosmological model. We first lay out the key physical processes in a primordial gas. Then, we introduce recent developments in computer simulations. Finally, we discuss prospects for future observations of the first generation of stars.

  2. Star Formation in Spiral Arms

    CERN Document Server

    Elmegreen, Bruce G

    2011-01-01

    The origin and types of spiral arms are reviewed with an emphasis on the connections between these arms and star formation. Flocculent spiral arms are most likely the result of transient instabilities in the gas that promote dense cloud formation, star formation, and generate turbulence. Long irregular spiral arms are usually initiated by gravitational instabilities in the stars, with the gas contributing to and following these instabilities, and star formation in the gas. Global spiral arms triggered by global perturbations, such as a galaxy interaction, can be wavemodes with wave reflection in the inner regions. They might grow and dominate the disk for several rotations before degenerating into higher-order modes by non-linear effects. Interstellar gas flows through these global arms, and through the more transient stellar spiral arms as well, where it can reach a high density and low shear, thereby promoting self-gravitational instabilities. The result is the formation of giant spiral arm cloud complexes,...

  3. A pore-forming toxin enables Serratia a nonlytic egress from host cells.

    Science.gov (United States)

    Di Venanzio, Gisela; Lazzaro, Martina; Morales, Enrique S; Krapf, Darío; García Véscovi, Eleonora

    2017-02-01

    Several pathogens co-opt host intracellular compartments to survive and replicate, and they thereafter disperse progeny to prosper in a new niche. Little is known about strategies displayed by Serratia marcescens to defeat immune responses and disseminate afterwards. Upon invasion of nonphagocytic cells, Serratia multiplies within autophagosome-like vacuoles. These Serratia-containing vacuoles (SeCV) circumvent progression into acidic/degradative compartments, avoiding elimination. In this work, we show that ShlA pore-forming toxin (PFT) commands Serratia escape from invaded cells. While ShlA-dependent, Ca(2)(+) local increase was shown in SeCVs tight proximity, intracellular Ca(2)(+) sequestration prevented Serratia exit. Accordingly, a Ca(2)(+) surge rescued a ShlA-deficient strain exit capacity, demonstrating that Ca(2)(+) mobilization is essential for egress. As opposed to wild-type-SeCV, the mutant strain-vacuole was wrapped by actin filaments, showing that ShlA expression rearranges host actin. Moreover, alteration of actin polymerization hindered wild-type Serratia escape, while increased intracellular Ca(2)(+) reorganized the mutant strain-SeCV actin distribution, restoring wild-type-SeCV phenotype. Our results demonstrate that, by ShlA expression, Serratia triggers a Ca(2)(+) signal that reshapes cytoskeleton dynamics and ends up pushing the SeCV load out of the cell, in an exocytic-like process. These results disclose that PFTs can be engaged in allowing bacteria to exit without compromising host cell integrity.

  4. Cyclosporine A prevents ex vivo PCO formation through induction of autophagy-mediated cell death.

    Science.gov (United States)

    Chandler, Heather L; Gervais, Kristen J; Lutz, Elizabeth A; Curto, Elizabeth M; Matusow, Rachel B; Wilkie, David A; Gemensky-Metzler, Anne J

    2015-05-01

    The purpose of this study was to determine the Cyclosporine A (CsA) dose and minimum drug delivery time needed to prevent posterior capsule opacification (PCO) in an ex vivo canine model and evaluate the mechanism of CsA-induced cell death. Canine lens epithelial cells (LEC) were treated with CsA and changes in cell migration, proliferation, and density were monitored over time. CsA-treated LEC underwent transmission electron microscopy (TEM), immunofluorescence, and immunoblotting in the presence or absence of autophagy inhibitors to evaluate the mechanism of cell death. Lens capsules were harvested from canine cadaver eyes for an ex vivo model of PCO. Lens capsules were treated with CsA for 1, 2, 3, 4, 5, 6, or 7 days, and subsequently maintained in culture for a total of 28 days in the absence of drug. CsA reduced LEC viability in a dose dependent manner. Morphologically, CsA-treated LEC were swollen, had intact nuclei, lacked peripheral chromatin condensation, and demonstrated prominent vacuolization; TEM revealed autophagosomes. LC3-II protein expression and acridine orange fluorescence increased in CsA-treated cells. A small non-significant induction of cleaved caspase-3 was observed in CsA-treated LEC. Lens capsules treated with 5, 6, or 7 days of 10 μg/mL CsA showed a significant decrease in ex vivo PCO formation; 6 days of drug delivery prevented PCO. This study finds that morphologic changes, formation of acidic vesicles, and increased expression of LC3-II supports the hypothesis that CsA mediates LEC death via autophagy; this is a novel finding in the lens. Induction of CsA-induced apoptosis was minimal. Six days of intracapsular CsA drug delivery prevented ex vivo PCO formation.

  5. Dwarf galaxy formation with H2-regulated star formation

    CERN Document Server

    Kuhlen, M; Madau, P; Smith, B; Wise, J

    2011-01-01

    We describe cosmological galaxy formation simulations with the adaptive mesh refinement code Enzo that incorporate a star formation prescription regulated by the local abundance of molecular hydrogen. We show that this H2-regulated prescription leads to a suppression of star formation in low mass halos (M_h 4, alleviating some of the dwarf galaxy problems faced by theoretical galaxy formation models. H2 regulation modifies the efficiency of star formation of cold gas directly, rather than indirectly reducing the cold gas content with "supernova feedback". We determine the local H2 abundance in our most refined grid cells (76 proper parsec in size at z=4) by applying the model of Krumholz, McKee, & Tumlinson, which is based on idealized 1D radiative transfer calculations of H2 formation-dissociation balance in ~100 pc atomic--molecular complexes. Our H2-regulated simulations are able to reproduce the empirical (albeit lower z) Kennicutt-Schmidt relation, including the low Sigma_gas cutoff due to the transi...

  6. AGN feedback in galaxy formation

    CERN Document Server

    Antonuccio-Delogu, Vincenzo

    2010-01-01

    During the past decade, convincing evidence has been accumulated concerning the effect of active galactic nuclei (AGN) activity on the internal and external environment of their host galaxies. Featuring contributions from well-respected researchers in the field, and bringing together work by specialists in both galaxy formation and AGN, this volume addresses a number of key questions about AGN feedback in the context of galaxy formation. The topics covered include downsizing and star-formation time scales in massive elliptical galaxies, the connection between the epochs of supermassive black h

  7. Free-format RPG IV

    CERN Document Server

    Martin, Jim

    2013-01-01

    This how-to guide offers a concise and thorough introduction to the increased productivity, better readability, and easier program maintenance that comes with the free-format style of programming in RPG IV. Although free-format information is available in IBM manuals, it is not separated from everything else, thereby requiring hours of tedious research to track down the information needed. This book provides everything one needs to know to write RPG IV in the free-format style, and author Jim Martin not only teaches rules and syntax but also explains how this new style of coding has the pot

  8. The multifaceted planetesimal formation process

    CERN Document Server

    Johansen, Anders; Tanaka, Hidekazu; Ormel, Chris; Bizzarro, Martin; Rickman, Hans

    2014-01-01

    Accumulation of dust and ice particles into planetesimals is an important step in the planet formation process. Planetesimals are the seeds of both terrestrial planets and the solid cores of gas and ice giants forming by core accretion. Left-over planetesimals in the form of asteroids, trans-Neptunian objects and comets provide a unique record of the physical conditions in the solar nebula. Debris from planetesimal collisions around other stars signposts that the planetesimal formation process, and hence planet formation, is ubiquitous in the Galaxy. The planetesimal formation stage extends from micrometer-sized dust and ice to bodies which can undergo run-away accretion. The latter ranges in size from 1 km to 1000 km, dependent on the planetesimal eccentricity excited by turbulent gas density fluctuations. Particles face many barriers during this growth, arising mainly from inefficient sticking, fragmentation and radial drift. Two promising growth pathways are mass transfer, where small aggregates transfer u...

  9. Understanding Galaxy Formation and Evolution

    CERN Document Server

    Avila-Reese, V

    2006-01-01

    The old dream of integrating into one the study of micro and macrocosmos is now a reality. Cosmology, astrophysics, and particle physics intersect in a scenario (but still not a theory) of cosmic structure formation and evolution called Lambda Cold Dark Matter (LCDM) model. This scenario emerged mainly to explain the origin of galaxies. In these lecture notes, I first present a review of the main galaxy properties, highlighting the questions that any theory of galaxy formation should explain. Then, the cosmological framework and the main aspects of primordial perturbation generation and evolution are pedagogically detached. Next, I focus on the ``dark side'' of galaxy formation, presenting a review on LCDM halo assembling and properties, and on the main candidates for non-baryonic dark matter. It is shown how the nature of elemental particles can influence on the features of galaxies and their systems. Finally, the complex processes of baryon dissipation inside the non-linearly evolving CDM halos, formation o...

  10. Portfolio Optimization under Habit Formation

    CERN Document Server

    Naryshkin, Roman

    2008-01-01

    The "standard" Merton formulation of optimal investment and consumption involves optimizing the integrated lifetime utility of consumption, suitably discounted, together with the discounted future bequest. In this formulation the utility of consumption at any given time depends only on the amount consumed at that time. However, it is both theoretically and empirically reasonable that an individuals utility of consumption would depend on past consumption history. Economists term this "Habit Formation". We introduce a new formulation of habit formation which allows non-addictive consumption patterns for a wide variety of utility specification. In this paper we construct a simple mathematical description of this habit formation and present numerical solutions. We compare the results with the standard ones and draw insights obtained from the habit formation. The consumption path tends to increase with time and be less sensitive to the market fluctuations, which perfectly reflects the existence of habit persistenc...

  11. Overview of Sequence Data Formats.

    Science.gov (United States)

    Zhang, Hongen

    2016-01-01

    Next-generation sequencing experiment can generate billions of short reads for each sample and processing of the raw reads will add more information. Various file formats have been introduced/developed in order to store and manipulate this information. This chapter presents an overview of the file formats including FASTQ, FASTA, SAM/BAM, GFF/GTF, BED, and VCF that are commonly used in analysis of next-generation sequencing data.

  12. First-order formative rules

    OpenAIRE

    Fuhs, Carsten; Kop, C.

    2014-01-01

    This paper discusses the method of formative rules for first-order term rewriting, which was previously defined for a higher-order setting. Dual to the well-known usable rules, formative rules allow dropping some of the term constraints that need to be solved during a termination proof. Compared to the higher-order definition, the first-order setting allows for significant improvements of the technique.

  13. IRIG Serial Time Code Formats

    Science.gov (United States)

    2016-08-01

    Standard This standard consists of a family of rate-scaled serial time codes with formats containing up to four coded expressions or words. All time...time code formats is designated A, B, D, E, G, and H. Various combinations of subwords and signal forms make up a time code word. To differentiate...ARE leap years. Additional information can be found at the following USNO web sites. • http://timeanddate.com/ date /leapyear.html • http

  14. Star Formation in Various Environments

    CERN Document Server

    Brosch, N; Spector, O; Zitrin, A

    2008-01-01

    We describe studies of star formation in various galaxies using primarily observations from the Wise Observatory. In addition to surface photometry in the broad band UBVRI, we also use a set of narrow-band H-alpha filters tuned to different redshifts to isolate the emission line. With these observational data, and using models of evolutionary stellar populations, we unravel the star formation histories of the galaxies and connect them to other parameters, such as the galaxy environment.

  15. Urothelial cultures support intracellular bacterial community formation by uropathogenic Escherichia coli.

    Science.gov (United States)

    Berry, Ruth E; Klumpp, David J; Schaeffer, Anthony J

    2009-07-01

    Uropathogenic Escherichia coli (UPEC) causes most community-acquired and nosocomial urinary tract infections (UTI). In a mouse model of UTI, UPEC invades superficial bladder cells and proliferates rapidly, forming biofilm-like structures called intracellular bacterial communities (IBCs). Using a gentamicin protection assay and fluorescence microscopy, we developed an in vitro model for studying UPEC proliferation within immortalized human urothelial cells. By pharmacologic manipulation of urothelial cells with the cholesterol-sequestering drug filipin, numbers of intracellular UPEC CFU increased 8 h and 24 h postinfection relative to untreated cultures. Enhanced UPEC intracellular proliferation required that the urothelial cells, but not the bacteria, be filipin treated prior to infection. However, neither UPEC frequency of invasion nor early intracellular trafficking events to a Lamp1-positive compartment were modulated by filipin. Upon inspection by fluorescence microscopy, cultures with enhanced UPEC intracellular proliferation exhibited large, dense bacterial aggregates within cells that resembled IBCs but were contained with Lamp1-positive vacuoles. While an isogenic fimH mutant was capable of forming these IBC-like structures, the mutant formed significantly fewer than wild-type UPEC. Similar to IBCs, expression of E. coli iron acquisition systems was upregulated by intracellular UPEC. Expression of other putative virulence factors, including hlyA, cnf1, fliC, kpsD, and the biofilm adhesin yfaL also increased, while expression of fimA decreased and that of flu did not change. These results indicate that UPEC differentially regulates virulence factors in the intracellular environment. Thus, immortalized urothelial cultures that recapitulate IBC formation in vitro represent a novel system for the molecular and biochemical characterization of the UPEC intracellular life cycle.

  16. Introduction to format: the software tools test formatting program

    Energy Technology Data Exchange (ETDEWEB)

    Agazzi, C.

    1984-12-01

    Format is the name of the Software Tools formatter. It allows you to format text according to instructions that you place within the text. The text and instructions for each document you wish to create are kept in files. Each instruction, called a request line, makes changes in the way your document is laid out. For example, you can change the margins within your document to visually set off lists of items or topics. You can also bold face or underline words or sentences to highlight them. Throughout this manual are examples of how to use the Format request lines along with illustrations of the effects request lines have on an example letter. The request lines begin with a period in the first column on the screen. Each request line performs a specific function and is placed on the line immediately in front of the test to be formatted. Output lines are automatically filled; that is, their right margins are justified, without regard to the format of the input test lines.

  17. Formative assessment in Swedish mathematics classroom practice

    OpenAIRE

    Andersson, Catarina; Boström, Erika

    2015-01-01

    International audience; Using formative assessment has woken interest in many countries because of the potential effect on student achievement. To investigate Swedish teachers' use of formative assessment in mathematics, this study used classroom observations and teacher interviews of 38 mathematics teachers. The teachers used formative assessment, but additional formative activities could support teachers to better take advantage of the potential in using formative assessment.

  18. Treating tar sands formations with karsted zones

    Energy Technology Data Exchange (ETDEWEB)

    Vinegar, Harold J. (Bellaire, TX); Karanikas, John Michael (Houston, TX)

    2010-03-09

    Methods for treating a tar sands formation are described herein. The tar sands formation may have one or more karsted zones. Methods may include providing heat from one or more heaters to one or more karsted zones of the tar sands formation to mobilize fluids in the formation. At least some of the mobilized fluids may be produced from the formation.

  19. Star formation and structure formation in galaxy collisions

    CERN Document Server

    Bournaud, Frederic

    2009-01-01

    A number of theoretical and simulation results on star and structure formation in galaxy interactions and mergers is reviewed, and recent hydrodynamic simulations are presented. The role of gravity torques and ISM turbulence in galaxy interactions, in addition to the tidal field, is highlighted. Interactions can drive gas inflows towards the central kpc and trigger a central starburst, the intensity and statistical properties of which are discussed. A kinematically decoupled core and a supermassive central black hole can be fueled. Outside of the central kpc, many structures can form inside tidal tails, collisional ring, bridges, including super star clusters and tidal dwarf galaxies. The formation of super star clusters in galaxy mergers can now be directly resolved in hydrodynamic simulations. Their formation mechanisms and long-term evolution are reviewed, and the connection with present-day early-type galaxies is discussed.

  20. The dynamics of latifundia formation.

    Science.gov (United States)

    Chaves, Luis Fernando

    2013-01-01

    Land tenure inequity is a major social problem in developing nations worldwide. In societies, where land is a commodity, inequities in land tenure are associated with gaps in income distribution, poverty and biodiversity loss. A common pattern of land tenure inequities through the history of civilization has been the formation of latifundia [Zhuāngyuán in chinese], i.e., a pattern where land ownership is concentrated by a small fraction of the whole population. Here, we use simple Markov chain models to study the dynamics of latifundia formation in a heterogeneous landscape where land can transition between forest, agriculture and recovering land. We systematically study the likelihood of latifundia formation under the assumption of pre-capitalist trade, where trade is based on the average utility of land parcels belonging to each individual landowner during a discrete time step. By restricting land trade to that under recovery, we found the likelihood of latifundia formation to increase with the size of the system, i.e., the amount of land and individuals in the society. We found that an increase of the transition rate for land use changes, i.e., how quickly land use changes, promotes more equitable patterns of land ownership. Disease introduction in the system, which reduced land profitability for infected individual landowners, promoted the formation of latifundia, with an increased likelihood for latifundia formation when there were heterogeneities in the susceptibility to infection. Finally, our model suggests that land ownership reforms need to guarantee an equitative distribution of land among individuals in a society to avoid the formation of latifundia.

  1. The dynamics of latifundia formation.

    Directory of Open Access Journals (Sweden)

    Luis Fernando Chaves

    Full Text Available Land tenure inequity is a major social problem in developing nations worldwide. In societies, where land is a commodity, inequities in land tenure are associated with gaps in income distribution, poverty and biodiversity loss. A common pattern of land tenure inequities through the history of civilization has been the formation of latifundia [Zhuāngyuán in chinese], i.e., a pattern where land ownership is concentrated by a small fraction of the whole population. Here, we use simple Markov chain models to study the dynamics of latifundia formation in a heterogeneous landscape where land can transition between forest, agriculture and recovering land. We systematically study the likelihood of latifundia formation under the assumption of pre-capitalist trade, where trade is based on the average utility of land parcels belonging to each individual landowner during a discrete time step. By restricting land trade to that under recovery, we found the likelihood of latifundia formation to increase with the size of the system, i.e., the amount of land and individuals in the society. We found that an increase of the transition rate for land use changes, i.e., how quickly land use changes, promotes more equitable patterns of land ownership. Disease introduction in the system, which reduced land profitability for infected individual landowners, promoted the formation of latifundia, with an increased likelihood for latifundia formation when there were heterogeneities in the susceptibility to infection. Finally, our model suggests that land ownership reforms need to guarantee an equitative distribution of land among individuals in a society to avoid the formation of latifundia.

  2. An Adaptable Seismic Data Format

    Science.gov (United States)

    Krischer, Lion; Smith, James; Lei, Wenjie; Lefebvre, Matthieu; Ruan, Youyi; de Andrade, Elliott Sales; Podhorszki, Norbert; Bozdağ, Ebru; Tromp, Jeroen

    2016-11-01

    We present ASDF, the Adaptable Seismic Data Format, a modern and practical data format for all branches of seismology and beyond. The growing volume of freely available data coupled with ever expanding computational power opens avenues to tackle larger and more complex problems. Current bottlenecks include inefficient resource usage and insufficient data organization. Properly scaling a problem requires the resolution of both these challenges, and existing data formats are no longer up to the task. ASDF stores any number of synthetic, processed or unaltered waveforms in a single file. A key improvement compared to existing formats is the inclusion of comprehensive meta information, such as event or station information, in the same file. Additionally, it is also usable for any non-waveform data, for example, cross-correlations, adjoint sources or receiver functions. Last but not least, full provenance information can be stored alongside each item of data, thereby enhancing reproducibility and accountability. Any data set in our proposed format is self-describing and can be readily exchanged with others, facilitating collaboration. The utilization of the HDF5 container format grants efficient and parallel I/O operations, integrated compression algorithms and check sums to guard against data corruption. To not reinvent the wheel and to build upon past developments, we use existing standards like QuakeML, StationXML, W3C PROV and HDF5 wherever feasible. Usability and tool support are crucial for any new format to gain acceptance. We developed mature C/Fortran and Python based APIs coupling ASDF to the widely used SPECFEM3D_GLOBE and ObsPy toolkits.

  3. 幽门螺杆菌VacA蛋白体外诱导胃上皮AGS细胞内HMGB1的表达%HMGB1 expression in gastric epithelial AGS cells in vitro induced by vacuolating cytotoxin of Helicobacter pylori

    Institute of Scientific and Technical Information of China (English)

    赵琪; 郭继中; 黄学文; 陈国千; 罗瑞华; 黄丽丽; 安仙园; 赵兰静

    2011-01-01

    目的 探讨幽门螺杆菌(HP)感染的胃上皮AGS细胞内高迁移率族蛋白B1(high mobbility group box 1,HMGB1)的表达.方法 Hpl1638(CagA+,VacA+)和Hp1638突变株(Hp1638M,CagA+,VacA-)的提取液与AGS细胞共同温育后,收集细胞及培养上清液.裂解AGS细胞,western blot分析AGS细胞内HMGB1的表达,ELISA法检测培养上清液中HMGB1的水平.结果 HpLL638提取液刺激AGS细胞后HMGB1表达量为(123.33±25.2)μg/mL,明显高于Hpll638M提取液刺激后的(46.67±7.23)μg/mL(q=8.49,P<0.01).Hp11638和Hp11638M提取液刺激的AGS细胞培养上清液中HMGB1的水平分别为(115.59±16.62)和(48.32±6.30)ng/mL,差异有统计学意义(q=12.25,P<0.01).结论 在胃炎发生、发展过程中,VacA蛋白是刺激细胞中HMGB1高表达的主要因子.%Objective To explore the expression of high mobility group box 1 (HMGB1) in gastric epithelial AGS cells infected by He-licobacter pylori (HP). Methods Both the extracts of Hp11638 strain, which was positively expressed cytotoxin-associated protein (CagA) and vacuolating cytotoxin (VacA) , and Hp] 1638 mutant strain (Hp11638M, CagA+ , VacA- ) were incubated with AGS cells respectively. The AGS cells and the supernatant were collected. The AGS cells were splitted to analyze the expression of HMGB1 by west-em blotting, and the level of HMGB1 in the supernatant was measured by ELISA. Results The content of HMGB1 in AGS cells infected by extracts of Hp11638 was (123.3 ±25.2) μg/mL, which was significantly higher than that of Hpll638M (46.67 ±7.23) μg/mL, q = 8.49, P <0.01. The level of HMGB1 in the culture supernatant infected by the extracts of Hpl 1638 was (115.59 ± 16. 62 ) ng/mL which was significantly higher than that of Hpll638M (48.32±6.30) ng/mL, q = 12. 25, P <0. 01. Conclusions During the development and advance of gastritis, vacuolating cytotoxin may be the main factor for stimulating AGS cells to highly express HMGB1 protein.

  4. Star formation in Galactic flows

    Science.gov (United States)

    Smilgys, Romas; Bonnell, Ian A.

    2016-06-01

    We investigate the triggering of star formation in clouds that form in Galactic scale flows as the interstellar medium passes through spiral shocks. We use the Lagrangian nature of smoothed particle hydrodynamics simulations to trace how the star-forming gas is gathered into self-gravitating cores that collapse to form stars. Large-scale flows that arise due to Galactic dynamics create shocks of the order of 30 km s-1 that compress the gas and form dense clouds (n > several × 102 cm-3) in which self-gravity becomes relevant. These large-scale flows are necessary for creating the dense physical conditions for gravitational collapse and star formation. Local gravitational collapse requires densities in excess of n > 103 cm-3 which occur on size scales of ≈1 pc for low-mass star-forming regions (M 103 M⊙). Star formation in the 250 pc region lasts throughout the 5 Myr time-scale of the simulation with a star formation rate of ≈10-1 M⊙ yr-1 kpc-2. In the absence of feedback, the efficiency of the star formation per free-fall time varies from our assumed 100 per cent at our sink accretion radius to values of <10-3 at low densities.

  5. VLBI Data Interchange Format (VDIF)

    Science.gov (United States)

    Whitney, Alan; Kettenis, Mark; Phillips, Chris; Sekido, Mamoru

    2010-01-01

    One important outcome of the 7th International e-VLBI Workshop in Shanghai in June 2008 was the creation of a task force to study and recommend a universal VLBI data format that is suitable for both on-the-wire e-VLBI data transfer, as well as direct disk storage. This task force, called the VLBI Data Interchange Format (VDIF) Task Force, is the first part of a two-part effort, the second of which will address standardization of e-VLBI data-transmission-protocols. The formation of the VDIF Task Force was prompted particularly by increased e-VLBI activity and the difficulties encountered when data arrive at a correlator in different formats from various instruments in various parts of the world. The task force created a streaming packetized data format that may be used for real-time and non-realtime e-VLBI, as well as direct disk storage. The data may contain multiple channels of time-sampled data with an arbitrary number of channels, arbitrary #bits/sample up to 32, and real or complex data; data rates in excess of 100 Gbps are supported. Each data packet is completely self-identifying via a short header, and data may be decoded without reference to any external information. The VDIF task force has completed its work, and the VDIF standard was ratified at the 2009 e-VLBI workshop in Madrid.

  6. Compulsory formation of solvento complexes

    Energy Technology Data Exchange (ETDEWEB)

    Kukushkin, Yu.N.; Kukushkin, V.Yu.

    1986-01-01

    The essence of the method of the compulsory formation of solvated complexes which are labile in substitution reactions has been described in this review. It is based on three techniques for eliminating ligands from the inner sphere of the original compounds: 1) binding of the ligands in sparingly soluble productions or their conversion into highly volatile compounds; 2) binding of the ligands in stronger compounds than the original products; 3) chemical conversion of coordinated ligands, which result in a loss of donor capacity by the reaction products and the departure of these products from the inner sphere of the complex compounds. If the solvent in which the reaction is carried out has a sufficient donor capability, the coordination capacity of the central ion is saturated by means of solvent molecules with the formation of a solvento complex. The method has been illustrated by a large number of concrete examples of reactions which result in the formation of compounds with an assigned composition.

  7. Black holes and galaxy formation

    CERN Document Server

    Propst, Raphael J

    2010-01-01

    Galaxies are the basic unit of cosmology. The study of galaxy formation is concerned with the processes that formed a heterogeneous universe from a homogeneous beginning. The physics of galaxy formation is complicated because it deals with the dynamics of stars, thermodynamics of gas and energy production of stars. A black hole is a massive object whose gravitational field is so intense that it prevents any form of matter or radiation to escape. It is hypothesized that the most massive galaxies in the universe- "elliptical galaxies"- grow simultaneously with the supermassive black holes at their centers, giving us much stronger evidence that black holes control galaxy formation. This book reviews new evidence in the field.

  8. Formation of porous gas hydrates

    CERN Document Server

    Salamatin, Andrey N

    2015-01-01

    Gas hydrates grown at gas-ice interfaces are examined by electron microscopy and found to have a submicron porous texture. Permeability of the intervening hydrate layers provides the connection between the two counterparts (gas and water molecules) of the clathration reaction and makes further hydrate formation possible. The study is focused on phenomenological description of principal stages and rate-limiting processes that control the kinetics of the porous gas hydrate crystal growth from ice powders. Although the detailed physical mechanisms involved in the porous hydrate formation still are not fully understood, the initial stage of hydrate film spreading over the ice surface should be distinguished from the subsequent stage which is presumably limited by the clathration reaction at the ice-hydrate interface and develops after the ice grain coating is finished. The model reveals a time dependence of the reaction degree essentially different from that when the rate-limiting step of the hydrate formation at...

  9. Bundle Formation in Biomimetic Hydrogels.

    Science.gov (United States)

    Jaspers, Maarten; Pape, A C H; Voets, Ilja K; Rowan, Alan E; Portale, Giuseppe; Kouwer, Paul H J

    2016-08-08

    Bundling of single polymer chains is a crucial process in the formation of biopolymer network gels that make up the extracellular matrix and the cytoskeleton. This bundled architecture leads to gels with distinctive properties, including a large-pore-size gel formation at very low concentrations and mechanical responsiveness through nonlinear mechanics, properties that are rarely observed in synthetic hydrogels. Using small-angle X-ray scattering (SAXS), we study the bundle formation and hydrogelation process of polyisocyanide gels, a synthetic material that uniquely mimics the structure and mechanics of biogels. We show how the structure of the material changes at the (thermally induced) gelation point and how factors such as concentration and polymer length determine the architecture, and with that, the mechanical properties. The correlation of the gel mechanics and the structural parameters obtained from SAXS experiments is essential in the design of future (synthetic) mimics of biopolymer networks.

  10. Dense Cloud Formation and Star Formation in a Barred Galaxy

    CERN Document Server

    Nimori, M; Sorai, K; Watanabe, Y; Hirota, A; Namekata, D

    2012-01-01

    We investigate the properties of massive, dense clouds formed in a barred galaxy and their possible relation to star formation, performing a two-dimensional hydrodynamical simulation with the gravitational potential obtained from the 2Mass data from the barred spiral galaxy, M83. Since the environment for cloud formation and evolution in the bar region is expected to be different from that in the spiral arm region, barred galaxies are a good target to study the environmental effects on cloud formation and the subsequent star formation. Our simulation uses for an initial 80 Myr an isothermal flow of non-self gravitating gas in the barred potential, then including radiative cooling, heating and self-gravitation of the gas for the next 40 Myr, during which dense clumps are formed. We identify many cold, dense gas clumps for which the mass is more than $10^4M_{\\odot}$ (a value corresponding to the molecular clouds) and study the physical properties of these clumps. The relation of the velocity dispersion of the i...

  11. Cytomorphologic significance of marginal vacuoles in diffuse thyroid enlargements

    Directory of Open Access Journals (Sweden)

    Anshu Gupta

    2013-01-01

    Conclusions: A significant association was found between abundant MVs and a hyperthyroid state. Moderate/absent MVs in diffuse goiters were not found to correlate with thyroid function. Thus, all diffuse goiters with prominent MVs require hormonal evaluation to rule out hyperfunction of the thyroid.

  12. Graphene Oxide Nanoribbons Induce Autophagic Vacuoles in Neuroblastoma Cell Lines

    Directory of Open Access Journals (Sweden)

    Emanuela Mari

    2016-11-01

    Full Text Available Since graphene nanoparticles are attracting increasing interest in relation to medical applications, it is important to understand their potential effects on humans. In the present study, we prepared graphene oxide (GO nanoribbons by oxidative unzipping of single-wall carbon nanotubes (SWCNTs and analyzed their toxicity in two human neuroblastoma cell lines. Neuroblastoma is the most common solid neoplasia in children. The hallmark of these tumors is the high number of different clinical variables, ranging from highly metastatic, rapid progression and resistance to therapy to spontaneous regression or change into benign ganglioneuromas. Patients with neuroblastoma are grouped into different risk groups that are characterized by different prognosis and different clinical behavior. Relapse and mortality in high risk patients is very high in spite of new advances in chemotherapy. Cell lines, obtained from neuroblastomas have different genotypic and phenotypic features. The cell lines SK-N-BE(2 and SH-SY5Y have different genetic mutations and tumorigenicity. Cells were exposed to low doses of GO for different times in order to investigate whether GO was a good vehicle for biological molecules delivering individualized therapy. Cytotoxicity in both cell lines was studied by measuring cellular oxidative stress (ROS, mitochondria membrane potential, expression of lysosomial proteins and cell growth. GO uptake and cytoplasmic distribution of particles were studied by Transmission Electron Microscopy (TEM for up to 72 h. The results show that GO at low concentrations increased ROS production and induced autophagy in both neuroblastoma cell lines within a few hours of exposure, events that, however, are not followed by growth arrest or death. For this reason, we suggest that the GO nanoparticle can be used for therapeutic delivery to the brain tissue with minimal effects on healthy cells.

  13. The changing nature of the Brucella-containing vacuole.

    Science.gov (United States)

    Celli, Jean

    2015-07-01

    Bacteria of the genus Brucella are intracellular vacuolar pathogens of mammals that cause the worldwide zoonosis brucellosis, and reside within phagocytes of infected hosts to promote their survival, persistence and proliferation. These traits are essential to the bacterium's ability to cause disease and have been the subject of much investigation to gain an understanding of Brucella pathogenic mechanisms. Although the endoplasmic reticulum-derived nature of the Brucella replicative niche has been long known, major strides have recently been made in deciphering the molecular mechanisms of its biogenesis, including the identification of bacterial determinants and host cellular pathways involved in this process. Here I will review and discuss the most recent advances in our knowledge of Brucella intracellular pathogenesis, with an emphasis on bacterial exploitation of the host endoplasmic reticulum-associated functions, and how autophagy-related processes contribute to the bacterium's intracellular cycle.

  14. Formation Criterion for Synthetic Jets

    Science.gov (United States)

    2005-10-01

    formation data for the axisymmetric case were published over 50 years ago by Ingard and Labate.10 More recent studies33,34 suggest that L0/d > 1 for...with the axisymmetric data from Ingard and Labate10 and Smith et al.33 are compared in Fig. 7. It is found that the available data are consis- tent with...the jet formation criterion with an empirically determined constant K equal to approximately 0.16. The deviation of Ingard and Labate’s data at their

  15. Magnetohydrodynamic mechanism for pedestal formation.

    Science.gov (United States)

    Guazzotto, L; Betti, R

    2011-09-16

    Time-dependent two-dimensional magnetohydrodynamic simulations are carried out for tokamak plasmas with edge poloidal flow. Differently from conventional equilibrium theory, a density pedestal all around the edge is obtained when the poloidal velocity exceeds the poloidal sound speed. The outboard pedestal is induced by the transonic discontinuity, the inboard one by mass redistribution. The density pedestal follows the formation of a highly sheared flow at the transonic surface. These results may be relevant to the L-H transition and pedestal formation in high performance tokamak plasmas.

  16. Star formation and gas supply

    Science.gov (United States)

    Catinella, B.

    2016-06-01

    A detailed knowledge of how gas cycles in and around galaxies, and how it depends on galaxy properties such as stellar mass and star formation rate, is crucial to understand galaxy formation and evolution. We take advantage of the most sensitive surveys of cold gas in massive galaxies, GASS and COLD GASS, as well as of the state-of-the-art HI blind survey ALFALFA to investigate how molecular and atomic hydrogen reservoirs vary along and across the main sequence of star-forming galaxies.

  17. Pattern formations and oscillatory phenomena

    CERN Document Server

    Kinoshita, Shuichi

    2013-01-01

    Patterns and their formations appear throughout nature, and are studied to analyze different problems in science and make predictions across a wide range of disciplines including biology, physics, mathematics, chemistry, material science, and nanoscience. With the emergence of nanoscience and the ability for researchers and scientists to study living systems at the biological level, pattern formation research has become even more essential. This book is an accessible first of its kind guide for scientists, researchers, engineers, and students who require a general introduction to thi

  18. Optimal Reconfiguration of Tetrahedral Formations

    Science.gov (United States)

    Huntington, Geoffrey; Rao, Anil V.; Hughes, Steven P.

    2004-01-01

    The problem of minimum-fuel formation reconfiguration for the Magnetospheric Multi-Scale (MMS) mission is studied. This reconfiguration trajectory optimization problem can be posed as a nonlinear optimal control problem. In this research, this optimal control problem is solved using a spectral collocation method called the Gauss pseudospectral method. The objective of this research is to provide highly accurate minimum-fuel solutions to the MMS formation reconfiguration problem and to gain insight into the underlying structure of fuel-optimal trajectories.

  19. Terrestrial Planet Formation: Constraining the Formation of Mercury

    Science.gov (United States)

    Lykawka, Patryk Sofia; Ito, Takashi

    2017-04-01

    How the four terrestrial planets of the solar system formed is one of the most fundamental questions in the planetary sciences. Particularly, the formation of Mercury remains poorly understood. We investigated terrestrial planet formation by performing 110 high-resolution N-body simulation runs using more than 100 embryos and 6000 disk planetesimals representing a primordial protoplanetary disk. To investigate the formation of Mercury, these simulations considered an inner region of the disk at 0.2–0.5 au (the Mercury region) and disks with and without mass enhancements beyond the ice line location, a IL, in the disk, where a IL = 1.5, 2.25, and 3.0 au were tested. Although Venus and Earth analogs (considering both orbits and masses) successfully formed in the majority of the runs, Mercury analogs were obtained in only nine runs. Mars analogs were also similarly scarce. Our Mercury analogs concentrated at orbits with a ∼ 0.27–0.34 au, relatively small eccentricities/inclinations, and median mass m ∼ 0.2 {M}\\oplus . In addition, we found that our Mercury analogs acquired most of their final masses from embryos/planetesimals initially located between 0.2 and ∼1–1.5 au within 10 Myr, while the remaining mass came from a wider region up to ∼3 au at later times. Although the ice line was negligible in the formation of planets located in the Mercury region, it enriched all terrestrial planets with water. Indeed, Mercury analogs showed a wide range of water mass fractions at the end of terrestrial planet formation.

  20. Galaxy Formation and SN Feedback

    CERN Document Server

    Tissera, P B; White, S D M; Springel, V

    2006-01-01

    We present a Supernova (SN) feedback model that succeeds at describing the chemical and energetic effects of SN explosions in galaxy formation simulations. This new SN model has been coupled to GADGET-2 and works within a new multiphase scheme which allows the description of a co-spatial mixture of cold and hot interstellar medium phases. No ad hoc scale-dependent parameters are associated to these SN and multiphase models making them particularly suited to studies of galaxy formation in a cosmological framework. Our SN model succeeds not only in setting a self-regulated star formation activity in galaxies but in triggering collimated chemical-enriched galactic winds. The effects of winds vary with the virial mass of the systems so that the smaller the galaxy, the larger the fraction of swept away gas and the stronger the decrease in its star formation activity. The fact that the fraction of ejected metals exceeds 60 per cent regardless of mass, suggests that SN feedback can be the responsible mechanism of th...

  1. The Road to Galaxy Formation

    CERN Document Server

    Keel, William C

    2007-01-01

    The formation of galaxies is one of the greatest puzzles in astronomy, the solution is shrouded in the depths of space and time, but has profound implications for the universe we observe today. The book discusses the beginnings of the process from cosmological observations and calculations, considers the broad features of galaxies that we need to explain and what we know of their later history. The author compares the competing theories for galaxy formation and considers the progress expected from new generations of powerful telescopes both on earth and in space. In this second edition the author has retained the observationally-based approach of the first edition, a feature which was particularly well-reviewed: Writing in Nature, Carlton Baugh noted in February 2003 that “It is refreshing, in a market dominated by theorists, to come across a book on galaxy formation written from an observational perspective. The Road to Galaxy Formation should prove to be a handy primer on observations for graduate student...

  2. Formative Evaluations in Online Classes

    Directory of Open Access Journals (Sweden)

    Jennifer Peterson

    2016-01-01

    Full Text Available Online courses are continuing to become an important component of higher education course offerings. As the number of such courses increases, the need for quality course evaluations and course improvements is also increasing. However, there is not general agreement on the best ways to evaluate and use evaluation data to improve online courses. While summative student evaluations are commonly used, these may not be the most effective for online course evaluation and quality improvement. Formative evaluation is one method of providing course evaluation and feedback to the instructor during the course while course improvements can be made to benefit the students currently in the course as well as future students. This method of evaluation not only provides for an effective course evaluation but also continuous improvement in the course. This case study reviews formative evaluations done in two online courses in a Health Information Management program. These formative evaluations were found to be quite effective for the identification and development of needed course improvement throughout the semester as well as for use as input into overall course evaluations. These evaluations demonstrate the value of formative evaluations and provide a methodology for implementing such evaluations into an online course.

  3. Electrochemical analysis of microdroplet formation

    NARCIS (Netherlands)

    Fukuyama, M.; Yoshida, Y.; Eijkel, J.C.T.; Berg, van den A.; Hibara, A.; Fujii, T.; Hibara, A.; Takeuchi, S.; Fukuba, T.

    2012-01-01

    This paper reports an electrochemical measurement system with a high-speed camera for observation of molecular transport phenomena at a water-oil (W/O) interface during microfluidic droplet formation. For demonstration of the system, currents corresponding to the transport of electrolyte ions to for

  4. Formation of Rationally Heterogeneous Expectations

    NARCIS (Netherlands)

    Pfajfar, D.

    2012-01-01

    Abstract: This paper models expectation formation by taking into account that agents produce heterogeneous expectations due to model uncertainty, informational frictions and different capacities for processing information. We show that there are two general classes of steady states within this frame

  5. Earth and Terrestrial Planet Formation

    CERN Document Server

    Jacobson, Seth A

    2015-01-01

    The growth and composition of Earth is a direct consequence of planet formation throughout the Solar System. We discuss the known history of the Solar System, the proposed stages of growth and how the early stages of planet formation may be dominated by pebble growth processes. Pebbles are small bodies whose strong interactions with the nebula gas lead to remarkable new accretion mechanisms for the formation of planetesimals and the growth of planetary embryos. Many of the popular models for the later stages of planet formation are presented. The classical models with the giant planets on fixed orbits are not consistent with the known history of the Solar System, fail to create a high Earth/Mars mass ratio, and, in many cases, are also internally inconsistent. The successful Grand Tack model creates a small Mars, a wet Earth, a realistic asteroid belt and the mass-orbit structure of the terrestrial planets. In the Grand Tack scenario, growth curves for Earth most closely match a Weibull model. The feeding zon...

  6. Inside-Out Planet Formation

    CERN Document Server

    Chatterjee, Sourav

    2013-01-01

    The compact multi-transiting planet systems discovered by Kepler challenge planet formation theory. Formation in situ from disks with radial mass surface density profiles similar to the minimum mass solar nebula (MMSN) but boosted in normalization by factors ~10 has been suggested. We propose that a more natural way to create these planets in the inner disk is formation sequentially from the inside-out via creation of successive gravitationally unstable rings fed from a continuous stream of small (~cm--m size) "pebbles", drifting inwards via gas drag. Pebbles collect at the pressure maximum associated with the transition from a magneto-rotational instability (MRI)-inactive ("dead zone") region to an inner MRI-active zone. A pebble ring builds up until it either becomes gravitationally unstable to form an ~1--10 M_\\Earth planet directly or induces gradual planet formation via core accretion. The planet continues to accrete from the disk until it becomes massive enough to isolate itself from the accretion flow....

  7. The EPRDATA Format: A Dialogue

    Energy Technology Data Exchange (ETDEWEB)

    Hughes, Henry Grady III [Los Alamos National Laboratory

    2015-08-18

    Recently the Los Alamos Nuclear Data Team has communicated certain issues of concern in relation to the new electron/photon/relaxation ACE data format as released in the eprdata12 library. In this document those issues are parsed, analyzed, and answered.

  8. The Formation of Galactic Bulges

    NARCIS (Netherlands)

    Peletier, R.; Balcells, M.; Falcon-Barroso, J.; Graham, A.

    2005-01-01

    We summarise some recent results about nearby galactic bulges that are relevant to their formation. We highlight a number of significant advances in our understanding of the surface brightness profiles, stellar populations, and especially the very centers of spiral galaxies. We also view our own Mil

  9. Genetic analysis of symbiosome formation

    NARCIS (Netherlands)

    Ovchinnikova, E.

    2012-01-01

    Endosymbiotic interactions form a fundament of life as we know it and are characterized by the formation of new specialized membrane compartments, in which the microbes are hosted inside living plant cells. A striking example is the symbiosis between legumes and nitrogen-fixing Rhizobium bacteria (r

  10. Mediating Among Diverse Data Formats.

    Science.gov (United States)

    1998-01-01

    different text editors often want to share documents, Microsoft Word programs permit users to load documents not only in Word’s own formats, but also in...HTML 47 0 DVI to PDF 42 0 Word to Postscript 39 2 error retrieving document Postscript to webbed images 25 1 ghostscript did not return I£ TEXto

  11. Formative Assessment in Dance Education

    Science.gov (United States)

    Andrade, Heidi; Lui, Angela; Palma, Maria; Hefferen, Joanna

    2015-01-01

    Feedback is crucial to students' growth as dancers. When used within the framework of formative assessment, or assessment for learning, feedback results in actionable next steps that dancers can use to improve their performances. This article showcases the work of two dance specialists, one elementary and one middle school teacher, who have…

  12. Exciton Formation in Disordered Semiconductors

    DEFF Research Database (Denmark)

    Klochikhin, A.; Reznitsky, A.; Permogorov, S.

    1999-01-01

    Stationary luminescence spectra of disordered solid solutions can be accounted by the model of localized excitons. Detailed analysis of the long time decay kinetics of luminescence shows that exciton formation in these systems is in great extent due to the bimolecular reaction of separated carrie...

  13. Control over Administrative Contract Formation

    Directory of Open Access Journals (Sweden)

    Frane Staničić

    2016-02-01

    Full Text Available Administrative contracts in Croatian legislation represent a novelty introduced into the General Administration Procedure Act in 2010. This is a novelty which has not proved to be successful in practice. Control over administrative contract formation is inevitable and is very significant for a number of reasons. Firstly, public legal bodies which form them do so by exercising their own public powers which are without doubt subject to legality control; secondly, in forming administrative contracts, public funds are used which must be controlled; thirdly, forming administrative contracts often touches on using public goods. Due to the restrictive interpretation of administrative contracts in Croatian legislation, this institute is indisputably only regulated in the General Taxation Act. However, for more than two decades contracts which satisfy all presumptions have existed in our law in order to be considered as administrative contracts. It is for this reason that control over contracts will be dealt with for contracts considered by the author to be administrative contracts. These are contracts on concessions and contracts on public procurement. How inadequate today’s regulation of control of administrative contract formation is will be demonstrated, particularly regarding contracts on concession and public procurement. Legislative changes will be proposed which should result in a more quality system of control over administrative contract formation. How control over administrative contract formation cannot be considered as separate from control over administrative contract execution will also be shown.

  14. Wintertime Haze Formation in Beijing

    Science.gov (United States)

    Levy Zamora, M. E.

    2015-12-01

    Recent severe haze events in China have attracted significant public attention due to the severely reduced visibility and unprecedentedly high pollutant concentrations. Particular attention has been given to the high concentrations of particulate matter with a diameter less than 2.5 microns (PM2.5), which can exceed several hundred micrograms per cubic meter over several days. During January and February of 2015, a suite of aerosol instruments was deployed in Beijing to directly measure a comprehensive set of aerosol properties, including the particle size distribution, effective density, and chemical composition. In this presentation, we will discuss the particulate matter formation mechanisms, the evolution of aerosol properties throughout the event, and how the winter formation mechanisms compare with the warmer seasons. We show that the periodic cycles of severe haze episodes in Beijing are largely driven by meteorological conditions. During haze events, stagnation typically develops as a result of a low planetary boundary layer and weak southerly wind from polluted industrial source regions. Stronger northerly winds were frequently observed during the clean period, which carry unpolluted air masses from the less populated northern mountainous areas. Nucleation consistently occurs on clean days, producing a high number concentration of nano particles. The particle mass concentration exceeding several hundred micrograms per cubic meter is attributed to the continuous size growth from the nucleation-mode particles (diameter less than 10 nm) over multiple days to produce a high concentration of larger particles (diameter greater than 100 nm). The particle chemical composition in Beijing is similar to those commonly measured in other urban centers, which is indicative of chemical constituents dominated by secondary aerosol formation. Our results reveal that the severe haze formation in Beijing during the wintertime is similar to the mechanism of haze formation

  15. Sedimentary pyrite formation: An update

    Science.gov (United States)

    Berner, Robert A.

    1984-04-01

    Sedimentary pyrite formation during early diagenesis is a major process for controlling the oxygen level of the atmosphere and the sulfate concentration in seawater over geologic time. The amount of pyrite that may form in a sediment is limited by the rates of supply of decomposable organic matter, dissolved sulfate, and reactive detrital iron minerals. Organic matter appears to be the major control on pyrite formation in normal (non-euxinic) terrigenous marine sediments where dissolved sulfate and iron minerals are abundant. By contrast, pyrite formation in non-marine, freshwater sediments is severely limited by low concentrations of sulfate and this characteristic can be used to distinguish ancient organic-rich fresh water shales from marine shales. Under marine euxinic conditions sufficient H 2S is produced that the dominant control on pyrite formation is the availability of reactive iron minerals. Calculations, based on a sulfur isotope model, indicate that over Phanerozoic time the worldwide average organic carbon-to-pyrite sulfur ratio of sedimentary rocks has varied considerably. High C/S ratios during Permo-Carboniferous time can be explained by a shift of major organic deposition from the oceans to the land which resulted in the formation of vast coal swamps at that time. Low C/S ratios, compared to today, during the early Paleozoic can be explained in terms of a greater abundance of euxinic basins combined with deposition of a more reactive type of organic matter in the remaining oxygenated portions of the ocean. The latter could have been due to lower oceanic oxygen levels and/or a lack of transportation of refractory terrestrial organic matter to the marine environment due to the absence of vascular land plants at that time.

  16. H$_2$-based star formation laws in galaxy formation models

    CERN Document Server

    Xie, Lizhi; Hirschmann, Michaela; Fontanot, Fabio; Zoldan, Anna

    2016-01-01

    We update our recently published model for GAlaxy Evolution and Assembly (GAEA), to include a self-consistent treatment of the partition of cold gas in atomic and molecular hydrogen. Our model provides significant improvements with respect to previous ones used for similar studies. In particular, GAEA (i) includes a sophisticated chemical enrichment scheme accounting for non-instantaneous recycling of gas, metals, and energy; (ii) reproduces the measured evolution of the galaxy stellar mass function; (iii) reproduces the observed correlation between galaxy stellar mass and gas metallicity at different redshifts. These are important prerequisites for models considering a metallicity dependent efficiency of molecular gas formation. We also update our model for disk sizes and show that model predictions are in nice agreement with observational estimates for the gas, stellar and star forming disks at different cosmic epochs. We analyse the influence of different star formation laws including empirical relations b...

  17. Standard Formats for Atomic Data: the APED

    Energy Technology Data Exchange (ETDEWEB)

    Smith, R K; Brickhouse, N S; Liedahl, D A; Raymond, J C

    2001-06-05

    Standardized formats for atomic data used in calculating emission from a collisionally-ionized plasma are described. The formats use the astronomical-standard FITS format, and are extendible to other purposes, such as photoionization data. The formats emphasize storing references to the original data source and keeping the data in as-received form, to aid in checking against the original literature.

  18. Treating tar sands formations with dolomite

    Energy Technology Data Exchange (ETDEWEB)

    Vinegar, Harold J.; Karanikas, John Michael

    2010-06-08

    Methods for treating a tar sands formation are described herein. The tar sands formation may include dolomite and hydrocarbons. Methods may include providing heat at less than the decomposition temperature of dolomite from one or more heaters to at least a portion of the formation. At least some of the hydrocarbon fluids are mobilized in the formation. At least some of the hydrocarbon fluids may be produced from the formation.

  19. Shock Formation in Lovelock Theories

    CERN Document Server

    Reall, Harvey S; Way, Benson

    2014-01-01

    We argue that Lovelock theories of gravity suffer from shock formation, unlike General Relativity. We consider the propagation of (i) a discontinuity in curvature, and (ii) weak, high frequency, gravitational waves. Such disturbances propagate along characteristic hypersurfaces of a "background" spacetime and their amplitude is governed by a transport equation. In GR the transport equation is linear. In Lovelock theories, it is nonlinear and its solutions can blow up, corresponding to the formation of a shock. We show that this effect is absent in some simple cases e.g. a flat background spacetime, and demonstrate its presence for a plane wave background. We comment on weak cosmic censorship, the evolution of shocks, and the nonlinear stability of Minkowski spacetime, in Lovelock theories.

  20. Complexity of formation in holography

    Science.gov (United States)

    Chapman, Shira; Marrochio, Hugo; Myers, Robert C.

    2017-01-01

    It was recently conjectured that the quantum complexity of a holographic boundary state can be computed by evaluating the gravitational action on a bulk region known as the Wheeler-DeWitt patch. We apply this complexity=action duality to evaluate the `complexity of formation' [1, 2], i.e. the additional complexity arising in preparing the entangled thermofield double state with two copies of the boundary CFT compared to preparing the individual vacuum states of the two copies. We find that for boundary dimensions d > 2, the difference in the complexities grows linearly with the thermal entropy at high temperatures. For the special case d = 2, the complexity of formation is a fixed constant, independent of the temperature. We compare these results to those found using the complexity=volume duality.

  1. Complexity of Formation in Holography

    CERN Document Server

    Chapman, Shira; Myers, Robert C

    2016-01-01

    It was recently conjectured that the quantum complexity of a holographic boundary state can be computed by evaluating the gravitational action on a bulk region known as the Wheeler-DeWitt patch. We apply this complexity=action duality to evaluate the `complexity of formation' (arXiv:1509.07876, arXiv:1512.04993), i.e., the additional complexity arising in preparing the entangled thermofield double state with two copies of the boundary CFT compared to preparing the individual vacuum states of the two copies. We find that for boundary dimensions $d>2$, the difference in the complexities grows linearly with the thermal entropy at high temperatures. For the special case $d=2$, the complexity of formation is a fixed constant, independent of the temperature. We compare these results to those found using the complexity=volume duality.

  2. New Paradigms For Asteroid Formation

    CERN Document Server

    Johansen, Anders; Cuzzi, Jeffrey N; Morbidelli, Alessandro; Gounelle, Matthieu

    2015-01-01

    Asteroids and meteorites provide key evidence on the formation of planetesimals in the Solar System. Asteroids are traditionally thought to form in a bottom-up process by coagulation within a population of initially km-scale planetesimals. However, new models challenge this idea by demonstrating that asteroids of sizes from 100 to 1000 km can form directly from the gravitational collapse of small particles which have organised themselves in dense filaments and clusters in the turbulent gas. Particles concentrate passively between eddies down to the smallest scales of the turbulent gas flow and inside large-scale pressure bumps and vortices. The streaming instability causes particles to take an active role in the concentration, by piling up in dense filaments whose friction on the gas reduces the radial drift compared to that of isolated particles. In this chapter we review new paradigms for asteroid formation and compare critically against the observed properties of asteroids as well as constraints from meteo...

  3. Star Cluster Formation and Feedback

    CERN Document Server

    Krumholz, Mark R; Arce, Hector G; Dale, James E; Gutermuth, Robert; Klein, Richard I; Li, Zhi-Yun; Nakamura, Fumitaka; Zhang, Qizhou

    2014-01-01

    Stars do not generally form in isolation. Instead, they form in clusters, and in these clustered environments newborn stars can have profound effects on one another and on their parent gas clouds. Feedback from clustered stars is almost certainly responsible for a number of otherwise puzzling facts about star formation: that it is an inefficient process that proceeds slowly when averaged over galactic scales; that most stars disperse from their birth sites and dissolve into the galactic field over timescales $\\ll 1$ Gyr; and that newborn stars follow an initial mass function (IMF) with a distinct peak in the range $0.1 - 1$ $M_\\odot$, rather than an IMF dominated by brown dwarfs. In this review we summarize current observational constraints and theoretical models for the complex interplay between clustered star formation and feedback.

  4. Formation of Supermassive Black Holes

    CERN Document Server

    Volonteri, Marta

    2010-01-01

    Evidence shows that massive black holes reside in most local galaxies. Studies have also established a number of relations between the MBH mass and properties of the host galaxy such as bulge mass and velocity dispersion. These results suggest that central MBHs, while much less massive than the host (~ 0.1%), are linked to the evolution of galactic structure. In hierarchical cosmologies, a single big galaxy today can be traced back to the stage when it was split up in hundreds of smaller components. Did MBH seeds form with the same efficiency in small proto-galaxies, or did their formation had to await the buildup of substantial galaxies with deeper potential wells? I briefly review here some of the physical processes that are conducive to the evolution of the massive black hole population. I will discuss black hole formation processes for `seed' black holes that are likely to place at early cosmic epochs, and possible observational tests of these scenarios.

  5. Coevolutionary modeling in network formation

    KAUST Repository

    Al-Shyoukh, Ibrahim

    2014-12-03

    Network coevolution, the process of network topology evolution in feedback with dynamical processes over the network nodes, is a common feature of many engineered and natural networks. In such settings, the change in network topology occurs at a comparable time scale to nodal dynamics. Coevolutionary modeling offers the possibility to better understand how and why network structures emerge. For example, social networks can exhibit a variety of structures, ranging from almost uniform to scale-free degree distributions. While current models of network formation can reproduce these structures, coevolutionary modeling can offer a better understanding of the underlying dynamics. This paper presents an overview of recent work on coevolutionary models of network formation, with an emphasis on the following three settings: (i) dynamic flow of benefits and costs, (ii) transient link establishment costs, and (iii) latent preferential attachment.

  6. Controlled Irradiative Formation of Penitentes

    CERN Document Server

    Bergeron, V; Betterton, M D

    2006-01-01

    Spike-shaped structures are produced by light-driven ablation in very different contexts. Penitentes 1-4 m high are common on Andean glaciers, where their formation changes glacier dynamics and hydrology. Laser ablation can produce cones 10-100 microns high with a variety of proposed applications in materials science. We report the first laboratory generation of centimeter-scale snow and ice penitentes. Systematically varying conditions allows identification of the essential parameters controlling the formation of ablation structures. We demonstrate that penitente initiation and coarsening requires cold temperatures, so that ablation leads to sublimation rather than melting. Once penitentes have formed, further growth of height can occur by melting. The penitentes intially appear as small structures (3 mm high) and grow by coarsening to 1-5 cm high. Our results are an important step towards understanding and controlling ablation morphologies.

  7. Photophoresis boosts giant planet formation

    CERN Document Server

    Teiser, Jens

    2013-01-01

    In the core accretion model of giant planet formation, a solid protoplanetary core begins to accrete gas directly from the nebula when its mass reaches about 5 earth masses. The protoplanet has at most a few million years to reach runaway gas accretion, as young stars lose their gas disks after 10 million years at the latest. Yet gas accretion also brings small dust grains entrained in the gas into the planetary atmosphere. Dust accretion creates an optically thick protoplanetary atmosphere that cannot efficiently radiate away the kinetic energy deposited by incoming planetesimals. A dust-rich atmosphere severely slows down atmospheric cooling, contraction, and inflow of new gas, in contradiction to the observed timescales of planet formation. Here we show that photophoresis is a strong mechanism for pushing dust out of the planetary atmosphere due to the momentum exchange between gas and dust grains. The thermal radiation from the heated inner atmosphere and core is sufficient to levitate dust grains and to ...

  8. Singularities formation, structure, and propagation

    CERN Document Server

    Eggers, J

    2015-01-01

    Many key phenomena in physics and engineering are described as singularities in the solutions to the differential equations describing them. Examples covered thoroughly in this book include the formation of drops and bubbles, the propagation of a crack and the formation of a shock in a gas. Aimed at a broad audience, this book provides the mathematical tools for understanding singularities and explains the many common features in their mathematical structure. Part I introduces the main concepts and techniques, using the most elementary mathematics possible so that it can be followed by readers with only a general background in differential equations. Parts II and III require more specialised methods of partial differential equations, complex analysis and asymptotic techniques. The book may be used for advanced fluid mechanics courses and as a complement to a general course on applied partial differential equations.

  9. Formation of quasiparallel Alfven solitons

    Science.gov (United States)

    Hamilton, R. L.; Kennel, C. F.; Mjolhus, E.

    1992-01-01

    The formation of quasi-parallel Alfven solitons is investigated through the inverse scattering transformation (IST) for the derivative nonlinear Schroedinger (DNLS) equation. The DNLS has a rich complement of soliton solutions consisting of a two-parameter soliton family and a one-parameter bright/dark soliton family. In this paper, the physical roles and origins of these soliton families are inferred through an analytic study of the scattering data generated by the IST for a set of initial profiles. The DNLS equation has as limiting forms the nonlinear Schroedinger (NLS), Korteweg-de-Vries (KdV) and modified Korteweg-de-Vries (MKdV) equations. Each of these limits is briefly reviewed in the physical context of quasi-parallel Alfven waves. The existence of these limiting forms serves as a natural framework for discussing the formation of Alfven solitons.

  10. Formation of polar ring galaxies

    CERN Document Server

    Bournaud, F

    2003-01-01

    Polar ring galaxies are peculiar systems in which a gas rich, nearly polar ring surrounds an early-type or elliptical host galaxy. Two formation scenarios for these objects have been proposed: they are thought to form either in major galaxy mergers or by tidal accretion of the polar material from a gas rich donor galaxy. Both scenarios are studied through N-body simulations including gas dynamics and star formation. Constraints on physical parameters are drawn out, in order to determine which scenario is the most likely to occur. Polar ring galaxies from each scenario are compared with observations and we discuss whether the accretion scenario and the merging scenario account for observational properties of polar ring galaxies. The conclusion of this study is that the accretion scenario is both the most likely and the most supported by observations. Even if the merging scenario is rather robust, most polar ring galaxies are shown to be the result of tidal gas accretion events.

  11. Cophasing the Planet Formation Imager

    CERN Document Server

    Petrov, Romain G; Elhalkouj, Thami; Monnier, John; Ireland, Michael; Kraus, Stefan

    2016-01-01

    The Planet Formation Imager (PFI) is a project for a very large optical interferometer intended to obtain images of the planet formation process at scales as small as the Hill sphere of giant exoplanets. Its main science instruments will work in the thermal infrared but it will be cophased in the near infrared, where it requires also some capacity for scientific imaging. PFI imaging and resolution specifications imply an array of 12 to 20 apertures and baselines up to a few kilometers cophased at near infrared coherent magnitudes as large as 10. This paper discusses various cophasing architectures and the corresponding minimum diameter of individual apertures, which is the dominant element of PFI cost estimates. From a global analysis of the possible combinations of pairwise fringe sensors, we show that conventional approaches used in current interferometers imply the use of prohibitively large telescopes and we indicate the innovative strategies that would allow building PFI with affordable apertures smaller...

  12. Galaxy formation through hierarchical clustering

    Science.gov (United States)

    White, Simon D. M.; Frenk, Carlos S.

    1991-01-01

    Analytic methods for studying the formation of galaxies by gas condensation within massive dark halos are presented. The present scheme applies to cosmogonies where structure grows through hierarchical clustering of a mixture of gas and dissipationless dark matter. The simplest models consistent with the current understanding of N-body work on dissipationless clustering, and that of numerical and analytic work on gas evolution and cooling are adopted. Standard models for the evolution of the stellar population are also employed, and new models for the way star formation heats and enriches the surrounding gas are constructed. Detailed results are presented for a cold dark matter universe with Omega = 1 and H(0) = 50 km/s/Mpc, but the present methods are applicable to other models. The present luminosity functions contain significantly more faint galaxies than are observed.

  13. Restucturing the Project Work Format

    DEFF Research Database (Denmark)

    Dupont, Søren

    2015-01-01

    The chapter is based on an evaluation of a pedagogical experiment at Roskilde University, the Antology Experiment. The objective of the experiment was to develop and expand the framework for project work through the production of anthologies compiled collectively by a number of project groups....... The novel aspects of the Anthology Experiment were most notably its magnitude and complexity. In this experiment the groups were totalling some 50 students who were working together. The experiment used a well-known publishing format from research, namely the anthology form, which usually focuses....... The organization was complex, the written product changed from project to article format, the students had to concentrate on working in their own project group as well as in the so-called clusters, and the form of exam was radically changed...

  14. Molecule Formation on Interstellar Grains

    Science.gov (United States)

    Vidali, G.

    2011-05-01

    The first experiments that were expressively designed to be applicable to hydrogen formation reactions in the ISM measured the efficiency of formation of molecular hydrogen on a polycrystalline olivine (Pirronello et al. (1997a)). It soon turned out that more was needed, and research began on the mechanism of reaction, on the in uence of the surface morphology, and on the excitation of the just- ormed molecule. In this review, I summarize what we learned from these and other experiments, and where more work is needed: in the elementary steps of reaction, in the bridging of the laboratory-ISM gap (large ux/large surface - small ux/small grain) using simulations, and in using realistic samples of dust grains. Understanding what experiments can and cannot deliver will help in designing and targeting observations, and vice-versa.

  15. The Chemistry of Planet Formation

    Science.gov (United States)

    Oberg, Karin I.

    2017-01-01

    Exo-planets are common, and they span a large range of compositions. The origins of the observed diversity of planetary compositions is largely unconstrained, but must be linked to the planet formation physics and chemistry. Among planets that are Earth-like, a second question is how often such planets form hospitable to life. A fraction of exo-planets are observed to be ‘physically habitable’, i.e. of the right temperature and bulk composition to sustain a water-based prebiotic chemistry, but this does not automatically imply that they are rich in the building blocks of life, in organic molecules of different sizes and kinds, i.e. that they are chemically habitable. In this talk I will argue that characterizing the chemistry of protoplanetary disks, the formation sites of planets, is key to address both the origins of planetary bulk compositions and the likelihood of finding organic matter on planets. The most direct path to constrain the chemistry in disks is to directly observe it. In the age of ALMA it is for the first time possible to image the chemistry of planet formation, to determine locations of disk snowlines, and to map the distributions of different organic molecules. Recent ALMA highlights include constraints on CO snowline locations, the discovery of spectacular chemical ring systems, and first detections of more complex organic molecules. Observations can only provide chemical snapshots, however, and even ALMA is blind to the majority of the chemistry that shapes planet formation. To interpret observations and address the full chemical complexity in disks requires models, both toy models and astrochemical simulations. These models in turn must be informed by laboratory experiments, some of which will be shown in this talk. It is thus only when we combine observational, theoretical and experimental constraints that we can hope to characterize the chemistry of disks, and further, the chemical compositions of nascent planets.

  16. Cosmological models of galaxy formation

    Science.gov (United States)

    Menci, N.

    I review the present status of galaxy formation models within a cosmological framework. I focus on semi-analytic models based on the Cold Dark Matter scenario, discussing the role of the different physical process involving dark matter and baryons in determining the observed statistical properties of galaxies and their dependence on cosmic time and on environment evolution. I will highlight some present problems and briefly present the main effects of assuming a Warm Dark Matter scenario.

  17. Star Formation in MUSCEL Galaxies

    Science.gov (United States)

    Young, Jason; Kuzio de Naray, Rachel; Wang, Sharon Xuesong

    2017-01-01

    We present preliminary star-formation histories for a subset of the low surface brightness (LSB) galaxies in the MUSCEL (MUltiwavelength observations of the Structure, Chemistry, and Evolution of LSB galaxies) program. These histories are fitted against ground-based IFU spectra in tandem with space-based UV and IR photometry. MUSCEL aims to use these histories along with kinematic analyses to determine the physical processes that have caused the evolution of LSB galaxies to diverge from their high surface brightness counterparts.

  18. Hydrodynamics of catheter biofilm formation

    CERN Document Server

    Sotolongo-Costa, Oscar; Rodriguez-Perez, Daniel; Martinez-Escobar, Sergio; Fernandez-Barbero, Antonio

    2009-01-01

    A hydrodynamic model is proposed to describe one of the most critical problems in intensive medical care units: the formation of biofilms inside central venous catheters. The incorporation of approximate solutions for the flow-limited diffusion equation leads to the conclusion that biofilms grow on the internal catheter wall due to the counter-stream diffusion of blood through a very thin layer close to the wall. This biological deposition is the first necessary step for the subsequent bacteria colonization.

  19. Effect of honokiol on Helicobacter pylori growth and the expression and activity of vacuolating cytotoxin A%和厚朴酚对幽门螺杆菌生长和空泡毒素A表达及活性的影响

    Institute of Scientific and Technical Information of China (English)

    刘丹; 廖顺花; 王莉新; 王易

    2013-01-01

    [目的]幽门螺杆菌(Helicobacter pylori,H.pylori)是胃炎和消化性溃疡的病原体,但其具有潜在的正常菌群的特性.本研究通过评价临床常用中药厚朴的活性成分和厚朴酚对H.pylori的抑制作用及对其空泡毒素A表达和活性的影响,以反映其对H.pylori具有的潜在去除毒性的作用.[方法]使用平皿稀释固体法和脑心浸液液体法测定和厚朴酚对H.pylori的最低抑菌浓度,进一步通过中性红摄入法评价经无抑菌效果的低浓度和厚朴酚干预后,H.pylori培养上清中空泡毒素A(Vacuolating cytotoxin A,VacA)的毒性作用;并通过RT-PCR和Western blot方法检测经低浓度和厚朴酚处理后,H.pylori菌体及分泌上清中VacA mRNA和蛋白的表达水平.[结果]发现高浓度(0.75 g/L)和厚朴酚对H.pylori具有抑制作用;而在远低于最低抑菌浓度时,和厚朴酚可有效抑制H.pylori VacA的形成和分泌.[结论]和厚朴酚具有下调H.pylori毒性的潜在作用,这为基于“致病性H.pylori的非致病性转变”这一机理的干预性治疗提供了有希望的研究前景.

  20. Illusory contour formation survives crowding.

    Science.gov (United States)

    Lau, Jonathan Siu Fung; Cheung, Sing-Hang

    2012-06-12

    Flanked objects are difficult to identify using peripheral vision due to visual crowding, which limits conscious access to target identity. Nonetheless, certain types of visual information have been shown to survive crowding. Such resilience to crowding provides valuable information about the underlying neural mechanism of crowding. Here we ask whether illusory contour formation survives crowding of the inducers. We manipulated the presence of illusory contours through the (mis)alignment of the four inducers of a Kanizsa square. In the inducer-aligned condition, the observers judged the perceived shape (thin vs. fat) of the illusory Kanizsa square, manipulated by small rotations of the inducers. In the inducer-misaligned condition, three of the four inducers (all except the upper-left) were rotated 90°. The observers judged the orientation of the upper-left inducer. Crowding of the inducers worsened observers' performance significantly only in the inducer-misaligned condition. Our findings suggest that information for illusory contour formation survives crowding of the inducers. Crowding happens at a stage where the low-level featural information is integrated for inducer orientation discrimination, but not at a stage where the same information is used for illusory contour formation.

  1. Hypothesis Formation, Paradigms, and Openness

    Directory of Open Access Journals (Sweden)

    Conrad P. Pritscher

    2008-01-01

    Full Text Available A part of hypothesis formation, while necessary for scientific investigation, is beyond direct observation. Powerful hypothesis formation is more than logical and is facilitated by mind­opening. As Percy Bridgeman, Nobel laureate, said, science is: “Nothing more than doing one's damnedest with one's mind, no holds barred.” This paper suggests more open schooling helps generate more open hypothesizing which helps one do one's damnedest with one's mind. It is hypothesized that a more open process of hypothesis formation may help schools and society forge new ways of living and learning so that more people more often can do their damnedest with their mind. This writing does not offer a new paradigm but rather attempts to elaborate on the notion that new paradigms are difficult to form without openness to what was previously quasi­unthinkable. More on these topics and issues is included in the author's Reopening Einstein's Thought: About What Can't Be Learned From Textbooks ­­to be published by Sense Publishers in June 2008.

  2. Kinetic competition during glass formation

    Energy Technology Data Exchange (ETDEWEB)

    Perepezko, J.H., E-mail: perepezk@engr.wisc.edu [University of Wisconsin-Madison, Department of Materials Science and Engineering, 1509 University Ave., Madison, WI 53706 (United States); Santhaweesuk, C.; Wang, J.Q. [University of Wisconsin-Madison, Department of Materials Science and Engineering, 1509 University Ave., Madison, WI 53706 (United States); Imhoff, S.D. [Los Alamos National Laboratory, Materials Science and Technology Div., Los Alamos, NM 87545 (United States)

    2014-12-05

    Highlights: • The kinetics of glass formation has been elucidated in an Fe and Au-base alloy. • A critical cooling rate range should be considered for glass formation. • Wedge casting, calorimetry and upquenching data are used to model TTT curves. - Abstract: For vitrification of an alloy melt during cooling there is a kinetic competition with the nucleation and growth of metastable and stable crystalline phases. Many of the measures of glass forming ability (GFA) attempt to capture some of the features of the kinetic competition, but the GFA metrics are static measures and the kinetic processes are dynamic in nature. In fact, the critical cooling rate for glass formation should be viewed in terms of a critical cooling rate range to acknowledge the stochastic nature of crystal nucleation behavior. Direct measurements of the critical cooling rate range confirm this behavior and also provide useful input for kinetics analysis. Usually kinetics analyses are based upon crystallization behavior that is measured either isothermally or upon heating to temperatures near the crystallization onset, T{sub x} and the results are extrapolated to much higher temperatures. This practice is based upon a number of assumptions about transport behavior in the undercooled liquid. With rapid up-quenching of amorphous samples, the high temperature crystallization behavior can be measured and used to refine the kinetics analysis and provide useful insight on the kinetic competition and glass forming ability.

  3. NSDF: Neuroscience Simulation Data Format.

    Science.gov (United States)

    Ray, Subhasis; Chintaluri, Chaitanya; Bhalla, Upinder S; Wójcik, Daniel K

    2016-04-01

    Data interchange is emerging as an essential aspect of modern neuroscience. In the areas of computational neuroscience and systems biology there are multiple model definition formats, which have contributed strongly to the development of an ecosystem of simulation and analysis tools. Here we report the development of the Neuroscience Simulation Data Format (NSDF) which extends this ecosystem to the data generated in simulations. NSDF is designed to store simulator output across scales: from multiscale chemical and electrical signaling models, to detailed single-neuron and network models, to abstract neural nets. It is self-documenting, efficient, modular, and scalable, both in terms of novel data types and in terms of data volume. NSDF is simulator-independent, and can be used by a range of standalone analysis and visualization tools. It may also be used to store variety of experimental data. NSDF is based on the widely used HDF5 (Hierarchical Data Format 5) specification and is open, platform-independent, and portable.

  4. Gravity, Turbulence, and Star Formation

    CERN Document Server

    Elmegreen, B G

    2004-01-01

    The azimuthal power spectra of optical emission from star formation and dust in spiral galaxies resembles the azimuthal power spectra of HI emission from the LMC. These and other power spectra of whole galaxies all resemble that of velocity in incompressible Kolmogorov turbulence. The reasons for this are unknown but it could be simply that star and cloud formation are the result of a mixture of processes and each gives a power spectrum similar to Kolmogorov turbulence, within the observable errors. The important point is that star and cloud formation are not random but are correlated over large distances by forces that span several orders of magnitude in scale. These forces are probably the usual combination of self-gravity, turbulence, and compression from stellar winds and supernovae, but they have to work in concert to create the structures we see in galaxies. In addition, the identification of flocculant spirals with swing amplified instabilities opens the possibility that a high fraction of turbulence i...

  5. Star Formation in Molecular Clouds

    CERN Document Server

    Krumholz, Mark R

    2011-01-01

    Star formation is one of the least understood processes in cosmic evolution. It is difficult to formulate a general theory for star formation in part because of the wide range of physical processes involved. The interstellar gas out of which stars form is a supersonically turbulent plasma governed by magnetohydrodynamics. This is hard enough by itself, since we do not understand even subsonic hydrodynamic turbulence very well, let alone supersonic non-ideal MHD turbulence. However, the behavior of star-forming clouds in the ISM is also obviously influenced by gravity, which adds complexity, and by both continuum and line radiative processes. Finally, the behavior of star-forming clouds is influenced by a wide variety of chemical processes, including formation and destruction of molecules and dust grains (which changes the thermodynamic behavior of the gas) and changes in ionization state (which alter how strongly the gas couples to magnetic fields). As a result of these complexities, there is nothing like a g...

  6. Positronium formation in various polyimides

    Energy Technology Data Exchange (ETDEWEB)

    Okamoto, Ken-ichi; Tanaka, Kazuhiro; Katsube, Mikio; Sueoka, Osamu; Ito, Yasuo (Yamaguchi Univ., Ube (Japan). Faculty of Engineering Tokyo Univ. (Japan). Research Center for Nuclear Science and Technology)

    1993-03-01

    Positronium (Ps) formation in various polyimides has been studied. It has been found that Ps yield is zero or small in the polyimides having pyromellitic dianhydride (PMDA) and 3,3[sup '],4,4[sup ']-benzophenonetetracarboxylic dianhydride (BTDA) as acid anhydride moiety, while those having 3,3[sup '],4,4[sup ']-biphenyltetracarboxylic dianhydride (BPDA) and 2,2-bis(3,4-dicarboxyphenyl)hexafluoropropane dianhydride (6FDA) form Ps with intensities up to about 20%. This difference is well correlated with the electron affinity of these moieties: PMDA > BTDA > BPDA [approx] 6FDA. In another experiment o-Ps yields and its lifetimes were measured in benzene solutions of monomeric model compounds (imide compounds prepared from n-butylamine and the acid anhydrides). It has been found that the model compounds from PMDA and BTDA both inhibit Ps formation and quench o-Ps lifetimes but that those from BPDA and 6FDA have neither the inhibition nor the quenching effects. The results show that the spur model is applicable for Ps formation in the polyimides. (Author).

  7. Positronium formation in various polyimides

    Science.gov (United States)

    Okamoto, Ken-ichi; Tanaka, Kazuhiro; Katsube, Mikio; Sueoka, Osamu; Ito, Yasuo

    1993-03-01

    Positronium (Ps) formation in various polyimides has been studied. It has been found that Ps yield is zero or small in the polyimides having pyromellitic dianhydride (PMDA) and 3,3',4,4'-benzophenonetetracarboxylic dianhydride (BTDA) as acid anhydride moiety, while those having 3,3',4,4'-biphenyltetracarboxylic dianhydride (BPDA) and 2,2- bis(3,4-dicarboxyphenyl)hexafluoropropane dianhydride (6FDA) form Ps with intensities up to about 20%. This difference is well correlated with the electron affinity of these moieties: PMDA > BTDA > BPDA ˜ 6FDA. In another experiment o-Ps yields and its lifetimes were measured in benzene solutions of monomeric model compounds (imide compounds prepared from n-butylamine and the acid anhydrides). It has been found that the model compounds from PMDA and BTDA both inhibit Ps formation and quench o-Ps lifetimes but that those from BPDA and 6FDA have neither the inhibition nor the quenching effects. The results show that the spur model is applicable for Ps formation in the polyimides.

  8. Galaxy Formation Spanning Cosmic History

    CERN Document Server

    Benson, Andrew J

    2010-01-01

    Over the past several decades, galaxy formation theory has met with significant successes. In order to test current theories thoroughly we require predictions for as yet unprobed regimes. To this end, we describe a new implementation of the Galform semi-analytic model of galaxy formation. Our motivation is the success of the model described by Bower et al. in explaining many aspects of galaxy formation. Despite this success, the Bower et al. model fails to match some observational constraints and certain aspects of its physical implementation are not as realistic as we would like. The model described in this work includes substantially updated physics, taking into account developments in our understanding over the past decade, and removes certain limiting assumptions made by this (and most other) semi-analytic models. This allows it to be exploited reliably in high-redshift and low mass regimes. Furthermore, we have performed an exhaustive search of model parameter space to find a particular set of model para...

  9. Star Formation in Satellite Galaxies

    CERN Document Server

    Gutíerrez, C M; Funes, J G; Ribeiro, M B

    2006-01-01

    We present narrow-band observations of the H$\\alpha$ emission in a sample of 31 satellite orbiting isolated giant spiral galaxies. The sample studied spans the range $-19formation rates are 0.68 and 3.66 M$_\\sun$ yr$^{-1}$ respectively. Maps of the spatial distribution of ionized gas are presented. The star-forming regions show a rich structure in which frequently discrete complexes are imposed over more diffuse structures. In general, the current star formation rates are smaller that the mean values in the past obtained from the current stellar content; this probably indicates a declining rhythm with time in the generation of new stars. However, the reserve of gas is enough to continue fueling the current levels of star formation activity for at least another Hubble time. Four of the o...

  10. Formation of Kuiper Belt Binaries

    CERN Document Server

    Goldreich, P; Sari, R; Goldreich, Peter; Lithwick, Yoram; Sari, Re'em

    2002-01-01

    It appears that at least several percent of large Kuiper belt objects are members of wide binaries. Physical collisions are too infrequent to account for their formation. Collisionless gravitational interactions are more promising. These provide two channels for binary formation. In each, the initial step is the formation of a transient binary when two large bodies penetrate each other's Hill spheres. Stabilization of a transient binary requires that it lose energy. Either dynamical friction due to small bodies or the scattering of a third large body can be responsible. Our estimates favor the former, albeit by a small margin. We predict that most objects of size comparable to those currently observed in the Kuiper belt are members of multiple systems. More specifically, we derive the probability that a large body is a member of a binary with semi-major axis of order a. The probability depends upon sigma, the total surface density, Sigma, the surface density of large bodies having radius R, and theta=10^-4, t...

  11. Interactions, Starbursts, and Star Formation

    Directory of Open Access Journals (Sweden)

    Johan H. Knapen

    2015-12-01

    Full Text Available We study how interactions between galaxies affect star formation within them by considering a sample of almost 1500 of the nearest galaxies, all within a distance of ∼45 Mpc. We use the far-IR emission to define the massive star formation rate (SFR, and then normalise the SFR by the stellar mass of the galaxy to obtain the specific star formation rate (SSFR. We explore the distribution of (SSFR with morphological type and with stellar mass. We calculate the relative enhancement of SFR and SSFR for each galaxy by normalising them by the median SFR and SSFR values of individual control samples of similar non-interacting galaxies. We find that both the median SFR and SSFR are enhanced in interacting galaxies, and more so as the degree of interaction is higher. The increase is moderate, reaching a maximum of a factor of 1.9 for the highest degree of interaction (mergers. While the SFR and SSFR are enhanced statistically by interactions, in many individual interacting galaxies they are not enhanced at all. Our study is based on a representative sample of nearby galaxies and should be used to place constraints on studies based on samples of galaxies at larger distances.

  12. Formation mechanisms of metal colloids

    Science.gov (United States)

    Halaciuga, Ionel

    Highly dispersed uniform metallic particles are widely used in various areas of technology and medicine and are likely to be incorporated into many other applications in the future. It is commonly accepted that size, shape and composition of the particles represent critical factors in most applications. Thus, understanding the mechanisms of formation of metal particles and the ways to control the physical (e.g. shape, size) and chemical (e.g. composition) properties is of great importance. In the current research, the formation of uniform silver spheres is investigated experimentally. The parameters that influence the formation of silver particles when concentrated iso-ascorbic acid and silver-polyamine complex solutions are rapidly mixed were studied in the absence of dispersants. We found that by varying the nature of the amine, temperature, concentration of reactants, silver/amine molar ratio, and the nature of the silver salt, the size of the resulting silver particles can be varied in a wide range (0.08--1.5 microm). The silver particles were formed by aggregation of nanosize subunits as substantiated by both electron microscopy and X-ray diffraction techniques and by the vivid rapid color changes during the chemical precipitation process. From the practical standpoint, the goal of this research was to prepare well dispersed spherical silver particles having a relatively smooth surface and a diameter of about 1 microm to satisfy the demands of the current electronic materials market. A two stage particle growth model previously developed to explain the narrow size distribution occurring in synthesis of gold spheres was applied to the present experimental system, and the parameters that control the size distribution characteristics were identified. The kinetic parameter required to match the final particle size was found to be in agreement with the one used previously in modeling formation of gold spheres, suggesting that similar kinetics governs the

  13. Core formation in silicate bodies

    Science.gov (United States)

    Nimmo, F.; O'Brien, D. P.; Kleine, T.

    2008-12-01

    Differentiation of a body into a metallic core and silicate mantle occurs most efficiently if temperatures are high enough to allow at least the metal to melt [1], and is enhanced if matrix deformation occurs [2]. Elevated temperatures may occur due to either decay of short-lived radio-isotopes, or gravitational energy release during accretion [3]. For bodies smaller than the Moon, core formation happens primarily due to radioactive decay. The Hf-W isotopic system may be used to date core formation; cores in some iron meteorites and the eucrite parent body (probably Vesta) formed within 1 My and 1-4~My of solar system formation, respectively [4]. These formation times are early enough to ensure widespread melting and differentiation by 26Al decay. Incorporation of Fe60 into the core, together with rapid early mantle solidification and cooling, may have driven early dynamo activity on some bodies [5]. Iron meteorites are typically depleted in sulphur relative to chondrites, for unknown reasons [6]. This depletion contrasts with the apparently higher sulphur contents of cores in larger planetary bodies, such as Mars [7], and also has a significant effect on the timing of core solidification. For bodies of Moon-size and larger, gravitational energy released during accretion is probably the primary cause of core formation [3]. The final stages of accretion involve large, stochastic collisions [8] between objects which are already differentiated. During each collision, the metallic cores of the colliding objects merge on timescales of a few hours [9]. Each collision will reset the Hf-W isotopic signature of both mantle and core, depending on the degree to which the impactor core re-equilibrates with the mantle of the target [10]. The re-equilibration efficiency depends mainly on the degree to which the impactor emulsifies [11], which is very uncertain. Results from N-body simulations [8,12] suggest that significant degrees of re- equilibration are required [4,10]. Re

  14. Clustering of Helicobacter pylori VacA in lipid rafts, mediated by its receptor, receptor-like protein tyrosine phosphatase beta, is required for intoxication in AZ-521 Cells

    DEFF Research Database (Denmark)

    Nakayama, Masaaki; Hisatsune, Jyunzo; Yamasaki, Eiki;

    2006-01-01

    Helicobacter pylori vacuolating cytotoxin, VacA, induces multiple effects on epithelial cells through different cellular events: one involves pore formation, leading to vacuolation, mitochondrial damage, and apoptosis, and the second involves cell signaling, resulting in stimulation of proinflamm......Helicobacter pylori vacuolating cytotoxin, VacA, induces multiple effects on epithelial cells through different cellular events: one involves pore formation, leading to vacuolation, mitochondrial damage, and apoptosis, and the second involves cell signaling, resulting in stimulation...

  15. MHD turbulence, cloud formation and star formation in the ISM

    CERN Document Server

    Vázquez-Semadeni, E; Pouquet, A

    1996-01-01

    We discuss the role of turbulence in cloud and star formation, as observed in numerical simulations of the interstellar medium. Turbulent compression at the interfaces of colliding gas streams is responsible for the formation of intermediate (\\simlt 100 pc) and small clouds (a few tens of pc), although the smallest clouds can also form from fragmentation of expanding shells around stellar heating centers. The largest cloud complexes (several hundred pc) seem to form by slow, gravitational instability-driven merging of individual clouds, which can actually be described as a large-scale tendency towards homogenization of the flow due to gravity rather than cloud collisions. These mechanisms operate as well in the presence of a magnetic field and rotation, although slight variations on the compressibility and cloud morphology are present which depend on the strength and topology of the field. In summary, the role of turbulence in the life-cycle of clouds appears to be twofold: small-scale modes contribute to clo...

  16. On-Going Galaxy Formation

    Science.gov (United States)

    Braine, Jonathan; Duc, P.-A.; Lisenfeld, U.; Charmandaris, V.; Vallejo, O.; Leon, S.; Brinks, E.

    2002-07-01

    We investigate the process of galaxy formation as can be observed in the only currently forming galaxies - the so-called Tidal Dwarf Galaxies, hereafter TDGs - through observations of the molecular gas detected via its CO (Carbon Monoxide) emission. These objects are formed of material torn off of the outer parts of a spiral disk due to tidal forces in a collision between two massive galaxies. Molecular gas is a key element in the galaxy formation process, providing the link between a cloud of gas and a bona fide galaxy. We have detected CO in 8 TDGs (Braine, Lisenfeld, Duc and Leon, 2000: Nature 403, 867; Braine, Duc, Lisenfeld, Charmandaris, Vallejo, Leon and Brinks: 2001, A&A 378, 51), with an overall detection rate of 80%, showing that molecular gas is abundant in TDGs, up to a few 108 M ⊙. The CO emission coincides both spatially and kinematically with the HI emission, indicating that the molecular gas forms from the atomic hydrogen where the HI column density is high. A possible trend of more evolved TDGs having greater molecular gas masses is observed, in accord with the transformation of HI into H2. Although TDGs share many of the properties of small irregulars, their CO luminosity is much greater (factor ˜ 100) than that of standard dwarf galaxies of comparable luminosity. This is most likely a consequence of the higher metallicity (≳sim 1/3 solar) of TDGs which makes CO a good tracer of molecular gas. This allows us to study star formation in environments ordinarily inaccessible due to the extreme difficulty of measuring the molecular gas mass. The star formation efficiency, measured by the CO luminosity per Hα flux, is the same in TDGs and full-sized spirals. CO is likely the best tracer of the dynamics of these objects because some fraction of the HI near the TDGs may be part of the tidal tail and not bound to the TDG. Although uncertainties are large for individual objects, as the geometry is unknown, our sample is now of eight detected objects

  17. DIKE FORMATION WITH SUBMERGED VANE

    Institute of Scientific and Technical Information of China (English)

    Umesh P.GUPTA; C.S.P.OJHA; Nayan SHARMA

    2006-01-01

    Submerged vanes are submerged foils constructed in a river at an angle of attack to induce horizontal circulation in the downstream flow. The initial height of vanes is 0.2 - 0.4 times the local water depth (d) at the design stage. Its length expressed as a multiple of the height to length ratio (H/L) normally is 0.25 to 0.5. With the use of a submerged vane in straight channels, dikes (heaps of sediment) were formed downstream. At an optimal angle of attack of 40o and at a Froude number of 0.25, the formation of the dike starts at some distance from the trailing edge of the rectangular and trapezoidal submerged vanes with a collar. A significant scour hole results at the trailing edge of a vane with a collar. A typical dike formed along the vane on the suction side with a symmetric bell-shaped profile. The dike formation phenomenon triggered by a vane close to the bank-line at a flow with a relatively higher Froude number, such as 0.25, does not seem to follow a straight alignment, but has a sort of curvilinear shape bent towards the suction side. The dike formation process for a flow with a smaller Froude number, such as 0.13, is observed to not be appreciable. With the same installation height of H, a vane without a collar was dislodged by the flow at a Froude number of 0.25, whereas it was stable at a Froude number of 0.13.

  18. Dark Matter and Galaxy Formation

    Science.gov (United States)

    Primack, Joel R.

    2009-12-01

    The four lectures that I gave in the XIII Ciclo de Cursos Especiais at the National Observatory of Brazil in Rio in October 2008 were (1) a brief history of dark matter and structure formation in a ΛCDM universe; (2) challenges to ΛCDM on small scales: satellites, cusps, and disks; (3) data on galaxy evolution and clustering compared with simulations; and (4) semi-analytic models. These lectures, themselves summaries of much work by many people, are summarized here briefly. The slides [1] contain much more information.

  19. Dark Matter and Galaxy Formation

    CERN Document Server

    Primack, Joel R

    2009-01-01

    The four lectures that I gave in the XIII Ciclo de Cursos Especiais at the National Observatory of Brazil in Rio in October 2008 were (1) a brief history of dark matter and structure formation in a LambdaCDM universe; (2) challenges to LambdaCDM on small scales: satellites, cusps, and disks; (3) data on galaxy evolution and clustering compared with simulations; and (4) semi-analytic models. These lectures, themselves summaries of much work by many people, are summarized here briefly.

  20. The formation of interstellar jets

    Science.gov (United States)

    Tenorio-Tagle, G.; Canto, J.; Rozyczka, M.

    1988-01-01

    The formation of interstellar jets by convergence of supersonic conical flows and the further dynamical evolution of these jets are investigated theoretically by means of numerical simulations. The results are presented in extensive graphs and characterized in detail. Strong radiative cooling is shown to result in jets with Mach numbers 2.5-29 propagating to lengths 50-100 times their original widths, with condensation of swept-up interstellar matter at Mach 5 or greater. The characteristics of so-called molecular outflows are well reproduced by the simulations of low-Mach-number and quasi-adiabatic jets.

  1. Massive neutrinos and galaxy formation

    CERN Document Server

    Ma Chung Pei

    1994-01-01

    We report the most recent results from high-resolution numerical simulations of structure formation in two flat cold+hot dark matter models with neutrino mass densities \\onu=0.2 and 0.3. We find that structure forms too late in all CDM+HDM models with \\onu>0.2 to account for the amount of dense neutral gas in high-redshift damped Lyman-\\alpha systems. The \\onu=0.2 model at z\\approx0 provides a better match to observations than the pure CDM model.

  2. Branch formation during organ development

    Science.gov (United States)

    Gjorevski, Nikolce; Nelson, Celeste M.

    2010-01-01

    Invertebrates and vertebrates use branching morphogenesis to build epithelial trees to maximize the surface area of organs within a given volume. Several molecular regulators of branching have recently been discovered, a number of which are conserved across different organs and species. Signals that control branching at the cellular and tissue levels are also starting to emerge, and are rapidly unveiling the physical nature of branch development. Here we discuss the molecular, cellular and physical processes that govern branch formation and highlight the major outstanding questions in the field. PMID:20890968

  3. Austenite formation during intercritical annealing

    OpenAIRE

    A. Lis; J. Lis

    2008-01-01

    Purpose: of this paper is the effect of the soft annealing of initial microstructure of the 6Mn16 steel on the kinetics of the austenite formation during next intercritical annealing.Design/methodology/approach: Analytical TEM point analysis with EDAX system attached to Philips CM20 was used to evaluate the concentration of Mn, Ni and Cr in the microstructure constituents of the multiphase steel and mainly Bainite- Martensite islands.Findings: The increase in soft annealing time from 1-60 hou...

  4. The multifaceted planetesimal formation process

    DEFF Research Database (Denmark)

    Johansen, Anders; Blum, Jürgen; Tanaka, Hidekazu

    2013-01-01

    Accumulation of dust and ice particles into planetesimals is an important step in the planet formation process. Planetesimals are the seeds of both terrestrial planets and the solid cores of gas and ice giants forming by core accretion. Left-over planetesimals in the form of asteroids, trans...... micrometer-sized dust and ice to bodies which can undergo run-away accretion. The latter ranges in size from 1 km to 1000 km, dependent on the planetesimal eccentricity excited by turbulent gas density fluctuations. Particles face many barriers during this growth, arising mainly from inefficient sticking...

  5. Formation flight astronomical survey telescope

    Science.gov (United States)

    Tsunemi, Hiroshi

    2012-03-01

    Formation Flight Astronomical Survey Telescope (FFAST) is a project for hard X-ray observation. It consists of two small satellites; one (telescope satellite) has a super mirror covering the energy range up to 80 keV while the other (detector satellite) has an scintillator deposited CCD (SDCCD) having good spatial resolution and high efficiency up to 100 keV. Two satellites will be put into individual Kepler orbits forming an X-ray telescope with a focal length of 20 m. They will be not in pointing mode but in survey mode to cover a large sky region.

  6. Quality guidance and quality formation

    DEFF Research Database (Denmark)

    Poulsen, Carsten Stig; Juhl, Hans Jørn; Kristensen, Kai

    1996-01-01

    This paper presents an extension of the quality guidance model of Steenkamp and van Trijp that includes consumer quality formation processes. Quality expectations and quality experiences are seen as antecedents of perceived overall product quality conceptual model is applied using LISREL to a data...... set on Danish butter cookies. Five plausible models of the relation between expectation, experience and perceived product quality are estimated. Finally one model is selected on the basis of three criteria: chi-square, RMSEA and AIC: The results show a model where expectations are indirectly related...... to perceived quality through experience. Udgivelsesdato: APR...

  7. Adsorption-induced step formation

    DEFF Research Database (Denmark)

    Thostrup, P.; Christoffersen, Ebbe; Lorensen, Henrik Qvist

    2001-01-01

    Through an interplay between density functional calculations, Monte Carlo simulations and scanning tunneling microscopy experiments, we show that an intermediate coverage of CO on the Pt(110) surface gives rise to a new rough equilibrium structure with more than 50% step atoms. CO is shown to bin...... so strongly to low-coordinated Pt atoms that it can break Pt-Pt bonds and spontaneously form steps on the surface. It is argued that adsorption-induced step formation may be a general effect, in particular at high gas pressures and temperatures....

  8. On the modelling of river delta formation

    NARCIS (Netherlands)

    Geleynse, N.

    2013-01-01

    This thesis presents approaches to the modelling of river delta formation. In particular, it provides results of numerical stratigraphic-morphodynamic modelling of river delta formation under various environmental forcings.

  9. FORMATE-BASED FLUIDS: FORMULATION AND APPLICATION

    Directory of Open Access Journals (Sweden)

    Nediljka Gaurina-Međimurec

    2008-12-01

    Full Text Available Formate-based fluids has been successfully used in over hunders HPHT well operations since they introduced in field practice. They have many advantages when compared with conventional HPHT drilling and completion fluids such as: minimal formation damage, maintenance of additve properties at high temperatures, reduced hydraulic flow resistance, low potential for differential sticking, naturally lubricating, very low corrosion rates, biodegradable and pose little risk to the environment etc. Formate-based fluids can be applied during deep slim hole drilling, shale drilling, reservoir drilling, salt and gas hydrate formations drilling. The laboratory research was carried out to evaluate the rheological behavior of formate-based fluids as a function of temperature. Formate-based fluids were formulated using potassium formate brine, xanthan polymer, PAC, starch and calcium carbonate. Experimental results show that potassium formate improves the thermal stability of polymers.

  10. XML Format for SESAME and LEOS

    Energy Technology Data Exchange (ETDEWEB)

    Durrenberger, J K; Neely, J R; Sterne, P A

    2009-04-29

    The objective of this document is to describe the XML format used by LLNL and LANL to represent the equation-of-state and related material information in the LEOS and SESAME data libraries. The primary purpose of this document is to describe a specific XML format for representing EOS data that is tailored to the nature of the underlying data and is amenable to conversion to both legacy SESAME and LEOS binary formats. The secondary purpose is to describe an XML format that lends itself to a 'natural' representation in a binary file format of the SESAME, pdb or hdf5 form so that this format and related tools can be used for the rapid and efficient development and implementation of prototype data structures. This document describes the XML format only. A working knowledge of LEOS and SESAME formats is assumed.

  11. Choosing to write the paper format thesis.

    Science.gov (United States)

    Morris, H M; Tipples, G

    1998-04-01

    Graduate students today may be faced with the option of writing either a traditional format thesis or a paper format thesis. In contrast to the traditional format in which the text body consists of four or five chapters, the body of the paper format thesis can be comprised of an introductory chapter, two or more papers written as publishable manuscripts, and a conclusion. In this article, an overview of the paper format thesis is presented and contrasted with the traditional format thesis. The description of the paper format thesis is followed by its advantages and disadvantages for writers and readers. It is by weighing all possible pros and cons, as well as considering one's individual situation, that the graduate student will be able to decide which format of thesis to write.

  12. The Formation of English Cyber- neologism

    Institute of Scientific and Technical Information of China (English)

    曹琪雯

    2014-01-01

    Based on the lexical word-building process, this paper firstly analyzes the characteristics of English cyber-neologism and then explores the formations of it in recent ten years, and finally discusses its influence on the formation of Chinese.

  13. Molecular mechanism of bone formation and regeneration

    Institute of Scientific and Technical Information of China (English)

    Akira Yamaguchi

    2008-01-01

    @@ Bone formation and regeneration are mediated by the coordinate action of various factors. Among these, bone morphogenetic protein (BMP) and runt-related gene 2 (Runx2) play crucial roles in bone formation.

  14. The Current Status of Galaxy Formation

    CERN Document Server

    Silk, Joe

    2012-01-01

    Understanding galaxy formation is one of the most pressing issues in cosmology. We review the current status of galaxy formation from both an observational and a theoretical perspective, and summarise the prospects for future advances.

  15. Tube Formation in Nanoscale Materials

    Directory of Open Access Journals (Sweden)

    Yan Chenglin

    2008-01-01

    Full Text Available Abstract The formation of tubular nanostructures normally requires layered, anisotropic, or pseudo-layered crystal structures, while inorganic compounds typically do not possess such structures, inorganic nanotubes thus have been a hot topic in the past decade. In this article, we review recent research activities on nanotubes fabrication and focus on three novel synthetic strategies for generating nanotubes from inorganic materials that do not have a layered structure. Specifically, thermal oxidation method based on gas–solid reaction to porous CuO nanotubes has been successfully established, semiconductor ZnS and Nb2O5nanotubes have been prepared by employing sacrificial template strategy based on liquid–solid reaction, and an in situ template method has been developed for the preparation of ZnO taper tubes through a chemical etching reaction. We have described the nanotube formation processes and illustrated the detailed key factors during their growth. The proposed mechanisms are presented for nanotube fabrication and the important pioneering studies are discussed on the rational design and fabrication of functional materials with tubular structures. It is the intention of this contribution to provide a brief account of these research activities.

  16. Radical formation by heavy metals

    Energy Technology Data Exchange (ETDEWEB)

    Beuter, W.

    1982-09-01

    Certain reduced heavy metal ions can convert oxygen to a ''reactive oxygen species'' by donation of an electron. The reactive oxygen then attacks structures susceptible to oxidation, in particular unsaturated fatty acids, and peroxidizes them in a radical reaction. This process is inhibited by the presence of vitamin E and by other means. Peroxidized lipids decay forming free radicals in the process which themselves can peroxidise neighbouring lipids in a radical chain reaction. This decay is, moreover, catalysed by reduced heavy metal ions but on the other hand retarded by selenium-containing glutathione peroxidase. Radical formation by heavy metals is considerably involved in (i) the production of parenteral iron poisoning of the piglet (ii) haemolytic crisis occurring in ruminants through chronic copper poisoning (iii) the production of lead poisoning in ruminants and other animals. These types of poisonings are made worse by a deficiency of vitamin E and/or selenium. Factors which increase the bio-availability of the free heavy metal ion or reduce the redox potential thereof can aid radical formation as well as factors which lead to a reduction of the heavy metal ion e.g. cysteine, ascorbic acid or glucose.

  17. Pattern formations and optimal packing.

    Science.gov (United States)

    Mityushev, Vladimir

    2016-04-01

    Patterns of different symmetries may arise after solution to reaction-diffusion equations. Hexagonal arrays, layers and their perturbations are observed in different models after numerical solution to the corresponding initial-boundary value problems. We demonstrate an intimate connection between pattern formations and optimal random packing on the plane. The main study is based on the following two points. First, the diffusive flux in reaction-diffusion systems is approximated by piecewise linear functions in the framework of structural approximations. This leads to a discrete network approximation of the considered continuous problem. Second, the discrete energy minimization yields optimal random packing of the domains (disks) in the representative cell. Therefore, the general problem of pattern formations based on the reaction-diffusion equations is reduced to the geometric problem of random packing. It is demonstrated that all random packings can be divided onto classes associated with classes of isomorphic graphs obtained from the Delaunay triangulation. The unique optimal solution is constructed in each class of the random packings. If the number of disks per representative cell is finite, the number of classes of isomorphic graphs, hence, the number of optimal packings is also finite.

  18. The Black Hole Formation Probability

    CERN Document Server

    Clausen, Drew; Ott, Christian D

    2014-01-01

    A longstanding question in stellar evolution is which massive stars produce black holes (BHs) rather than neutron stars (NSs) upon death. It has been common practice to assume that a given zero-age main sequence (ZAMS) mass star (and perhaps a given metallicity) simply produces either an NS or a BH, but this fails to account for a myriad of other variables that may effect this outcome, such as spin, binarity, or even stochastic differences in the stellar structure near core collapse. We argue that instead a probabilistic description of NS versus BH formation may be better suited to account for the current uncertainties in understanding how massive stars die. Using the observed BH mass distribution from Galactic X-ray binaries, we derive the probability that a star will make a BH as a function of its ZAMS mass, $P_{\\rm BH}(M_{\\rm ZAMS})$. We explore possible biases in the observed BH mass distribution and find that this sample is best suited for studying BH formation in stars with ZAMS masses in the range $12-...

  19. Simulations for terrestrial planets formation

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    In this paper,the formation of terrestrial planets in the late stage of planetary formation is investigated using the two-planet model.At that time,the protostar formed for about 3 Ma and the gas disk dissipated.In the model,the perturbations from Jupiter and Saturn are considered.Variations of the mass of outer planet,and the initial eccentricities and inclinations of embryos and planetesimals are also considered.Our results show that,terrestrial planets are formed in 50 Ma,and the accretion rate is about 60%-80%.In each simulation,3-4 terrestrial planets are formed inside"Jupiter"with masses of 0.15 -3.6M⊕.In the 0.5-4 AU,when the eccentricities of planetesimals are excited,planetesimals are able to accrete material from wide radial direction.The plenty of water material of the terrestrial planet in the Habitable Zone may be transferred from the farther places by this mechanism.Accretion could also happen a few times between two major planets only if the outer planet has a moderate mass and the small terrestrial planet could survive at some resonances over time scale of 10 8 a.In one of our simulations,commensurability of the orbital periods of planets is very common.Moreover,a librating-circulating 3:2 configuration of mean motion resonance is found.

  20. Simulations for terrestrial planets formation

    Institute of Scientific and Technical Information of China (English)

    ZHANG Niu; JI JiangHui

    2009-01-01

    In this paper, the formation of terrestrial planets in the late stage of planetary formation is Investigated using the two-planet model. At that time, the protostar formed for about 3 Ma and the gas disk dissipated. In the model, the perturbations from Jupiter and Saturn are considered. Variations of the mass of outer planet, and the initial eccentricities and inclinations of embryos and planetesimals are also considered. Our results show that, terrestrial planets are formed in 50 Ma, and the accretion rate is about 60%-80%. In each simulation, 3-4 terrestrial planets are formed inside "Jupiter" with masses of 0.15-3.6 M(⊙). In the 0.5-4 AU, when the eccentricities of planetesimals are excited, planetesimals are able to accrete material from wide radial direction. The plenty of water material of the terrestrial planet in the Habitable Zone may be transferred from the farther places by this mechanism. Accretion could also happen a few times between two major planets only if the outer planet has a moderate mass and the small terrestrial planet could survive at some resonances over time scale of 108a. In one of our simulations, commensurability of the orbital periods of planets is very common. Moreover, a librating-circulating 3:2 configuration of mean motion resonance is found.

  1. Characteristic Scales in Galaxy Formation

    CERN Document Server

    Dekel, A

    2004-01-01

    Recent data, e.g. from SDSS and 2dF, reveal a robust bi-modality in the distribution of galaxy properties, with a characteristic transition scale at stellar mass M_*~3x10^{10} Msun (near L_*), corresponding to virial velocity V~100 km/s. Smaller galaxies tend to be blue disks of young populations. They define a "fundamental line" of decreasing surface brightness, metallicity and velocity with decreasing M_*, which extends to the smallest dwarf galaxies. Galaxies above the critical scale are dominated by red spheroids of old populations, with roughly constant high surface brightens and metallicity, and they tend to host AGNs. A minimum in the virial M/L is obtained at the same magic scale. This bi-modality can be the combined imprint of several different physical processes. On smaller scales, disks are built by cold flows, and supernova feedback is effective in regulating star formation. On larger scales, the infalling gas is heated by a virial shock and star formation can be suppressed by AGN feedback. Anothe...

  2. The Formation of Galactic Disks

    CERN Document Server

    Mo, H J; White, S D M; Mao, Shude; White, Simon D.M.

    1997-01-01

    We study the population of galactic disks expected in current hierarchical clustering models for structure formation. A rotationally supported disk with exponential surface density profile is assumed to form with a mass and angular momentum which are fixed fractions of those of its surrounding dark halo. We assume that haloes respond adiabatically to disk formation, and that only stable disks can correspond to real systems. With these assumptions the predicted population can match both present-day disks and the damped Lyman alpha absorbers in QSO spectra. Good agreement is found provided: (i) the masses of disks are a few percent of those of their haloes; (ii) the specific angular momenta of disks are similar to those of their haloes; (iii) present-day disks were assembled recently (at z3kpc/h and about 10% at r>10kpc/h. The cross-section for absorption is strongly weighted towards disks with large angular momentum and so large size for their mass. The galaxy population associated with damped absorbers should...

  3. The formation of flashbulb memories.

    Science.gov (United States)

    Conway, M A; Anderson, S J; Larsen, S F; Donnelly, C M; McDaniel, M A; McClelland, A G; Rawles, R E; Logie, R H

    1994-05-01

    A large group of subjects took part in a multinational test-retest study to investigate the formation of flashbulb (FB) memories for learning the news of the resignation of the British prime minister, Margaret Thatcher. Over 86% of the U.K. subjects were found to have FB memories nearly 1 year after the resignation; their memory reports were characterized by spontaneous, accurate, and full recall of event details, including minutiae. In contrast, less than 29% of the non-U.K. subjects had FB memories 1 year later; memory reports in this group were characterized by forgetting, reconstructive errors, and confabulatory responses. A causal analysis of secondary variables showed that the formation of FB memories was primarily associated with the level of importance attached to the event and level of affective response to the news. These findings lend some support to the study by R. Brown and Kulik (1977), who suggest that FB memories may constitute a class of autobiographical memories distinguished by some form of preferential encoding.

  4. Enamel formation and amelogenesis imperfecta.

    Science.gov (United States)

    Hu, Jan C-C; Chun, Yong-Hee P; Al Hazzazzi, Turki; Simmer, James P

    2007-01-01

    Dental enamel is the epithelial-derived hard tissue covering the crowns of teeth. It is the most highly mineralized and hardest tissue in the body. Dental enamel is acellular and has no physiological means of repair outside of the protective and remineralization potential provided by saliva. Enamel is comprised of highly organized hydroxyapatite crystals that form in a defined extracellular space, the contents of which are supplied and regulated by ameloblasts. The entire process is under genetic instruction. The genetic control of amelogenesis is poorly understood, but requires the activities of multiple components that are uniquely important for dental enamel formation. Amelogenesis imperfecta (AI) is a collective designation for the variety of inherited conditions displaying isolated enamel malformations, but the designation is also used to indicate the presence of an enamel phenotype in syndromes. Recently, genetic studies have demonstrated the importance of genes encoding enamel matrix proteins in the etiology of isolated AI. Here we review the essential elements of dental enamel formation and the results of genetic analyses that have identified disease-causing mutations in genes encoding enamel matrix proteins. In addition, we provide a fresh perspective on the roles matrix proteins play in catalyzing the biomineralization of dental enamel.

  5. Formation number for vortex dipoles

    Science.gov (United States)

    Sadri, Vahid; Krueger, Paul S.

    2016-11-01

    This investigation considers the axisymmetric formation of two opposite sign concentric vortex rings from jet ejection between concentric cylinders. This arrangement is similar to planar flow in that the vortex rings will travel together when the gap between the cylinders is small, similar to a vortex dipole, but it has the advantage that the vortex motion is less constrained than the planar case (vortex stretching and vortex line curvature is allowed). The flow was simulated numerically at a jet Reynolds number of 1,000 (based on ΔR and the jet velocity), jet pulse length-to-gap ratio (L / ΔR) in the range 10-20, and gap-to-outer radius ratio (ΔR /Ro) in the range 0.01-0.1. Small gap ratios were chosen for comparison with 2D results. In contrast with 2D results, the closely paired vortices in this study exhibited pinch-off from the generating flow and finite formation numbers. The more complex flow evolution afforded by the axisymmetric model and its influence on the pinch-off process will be discussed. This material is based on work supported by the National Science Foundation under Grant No. 1133876 and SMU. This supports are gratefully acknowledged.

  6. Treating nahcolite containing formations and saline zones

    Energy Technology Data Exchange (ETDEWEB)

    Vinegar, Harold J

    2013-06-11

    A method for treating a nahcolite containing subsurface formation includes removing water from a saline zone in or near the formation. The removed water is heated using a steam and electricity cogeneration facility. The heated water is provided to the nahcolite containing formation. A fluid is produced from the nahcolite containing formation. The fluid includes at least some dissolved nahcolite. At least some of the fluid is provided to the saline zone.

  7. String Formatting Considered Harmful for Novice Programmers

    Science.gov (United States)

    Hughes, Michael C.; Jadud, Matthew C.; Rodrigo, Ma. Mercedes T.

    2010-01-01

    In Java, "System.out.printf" and "String.format" consume a specialised kind of string commonly known as a format string. In our study of first-year students at the Ateneo de Manila University, we discovered that format strings present a substantial challenge for novice programmers. Focusing on their first laboratory we found that 8% of all the…

  8. Spiritual Formation within Christian Higher Education

    Science.gov (United States)

    Otto, Patrick; Harrington, Michael

    2016-01-01

    The Christian university has a distinct responsibility to provide an environment devoted to the spiritual formation of students. Spiritual formation is not to be viewed as the only important goal of the university, thereby sacrificing intellectual or relational development, but rather spiritual formation is the aggregate "product" of the…

  9. Formation of tax culture in Russia

    Directory of Open Access Journals (Sweden)

    Halikova Je.A.

    2014-11-01

    Full Text Available This article deals with the mechanism of the formation of tax culture in Russia, moral and ethical principles, on which based the work of the tax authorities, given the author's idea of the formation of tax culture. We consider the institution of tax advice, its interaction with the tax authorities and its impact on the formation of tax culture.

  10. Bubble Formation in Silicon-Quartz Interface

    OpenAIRE

    Kakimoto, K.; EGUCHI, M.; Ozoe, H.

    1997-01-01

    Bubble formation at an interface between silicon melt and a quartz crucible was studied by thermodynamical calculation and visualization of bubble formation using X-ray radiography. A phase diagram of silicon-oxygen (Si-O) system is also calculated from the reported thermodynamical data. Critical temperature and radius of bubble formation at the interface was discussed.

  11. Diet History Questionnaire II: Size Formats

    Science.gov (United States)

    Two serving size formats are used on the NCI versions of the DHQ as shown below. Format 1 is used for nearly all serving size questions. Format 2 is used only in special cases, where 'never' is allowed as a response.

  12. Successful Student Writing through Formative Assessment

    Science.gov (United States)

    Tuttle, Harry Grover

    2010-01-01

    Use formative assessment to dramatically improve your students' writing. In "Successful Student Writing Through Formative Assessment", educator and international speaker Harry G. Tuttle shows you how to guide middle and high school students through the prewriting, writing, and revision processes using formative assessment techniques that work.…

  13. Formative Assessment: Guidance for Early Childhood Policymakers

    Science.gov (United States)

    Riley-Ayers, Shannon

    2014-01-01

    This policy report provides a guide and framework to early childhood policymakers considering formative assessment. The report defines formative assessment and outlines its process and application in the context of early childhood. The substance of this document is the issues for consideration in the implementation of the formative assessment…

  14. Formative Assessment: Responding to Your Students

    Science.gov (United States)

    Tuttle, Harry Grover

    2009-01-01

    This "how-to" book on formative assessment is filled with practical suggestions for teachers who want to use formative assessment in their classrooms. With practical strategies, tools, and examples for teachers of all subjects and grade levels, this book shows you how to use formative assessment to promote successful student learning. Topics…

  15. A new PICL trace file format

    Energy Technology Data Exchange (ETDEWEB)

    Worley, P.H.

    1992-10-01

    A trace file format is described that will be used in future releases of the Portable Instrumented Communication Library (PICL) and ParaGraph. The new format provides improved support for tracing and profiling PICL communication primitives and user-defined events. The new format is also easily extended and may be useful in other instrumentation packages and performance visualization tools.

  16. Supermassive Black Holes and Galaxy Formation

    OpenAIRE

    Silk, Joseph

    2001-01-01

    The formation of supermassive black holes (SMBH) is intimately related to galaxy formation, although precisely how remains a mystery. I speculate that formation of, and feedback from, SMBH may alleviate problems that have arisen in our understanding of the cores of dark halos of galaxies.

  17. Theoretical Considerations of Massive Star Formation

    Science.gov (United States)

    Yorke, Harold W.

    2006-01-01

    This viewgraph presentation reviews the formation of massive stars. The formation of massive stars is different in many ways from the formation of other stars. The presentation shows the math, and the mechanisms that must be possible for a massive star to form.

  18. Mud Volcanoes Formation And Occurrence

    Science.gov (United States)

    Guliyev, I. S.

    2007-12-01

    Mud volcanoes are natural phenomena, which occur throughout the globe. They are found at a greater or lesser scale in Azerbaijan, Turkmenistan, Georgia, on the Kerch and Taman peninsulas, on Sakhalin Island, in West Kuban, Italy, Romania, Iran, Pakistan, India, Burma, China, Japan, Indonesia, Malaysia, New Zealand, Mexico, Colombia, Trinidad and Tobago, Venezuela and Ecuador. Mud volcanoes are most well-developed in Eastern Azerbaijan, where more than 30% of all the volcanoes in the world are concentrated. More than 300 mud volcanoes have already been recognized here onshore or offshore, 220 of which lie within an area of 16,000 km2. Many of these mud volcanoes are particularly large (up to 400 m high). The volcanoes of the South Caspian form permanent or temporary islands, and numerous submarine banks. Many hypotheses have been developed regarding the origin of mud volcanoes. Some of those hypotheses will be examined in the present paper. Model of spontaneous excitation-decompaction (proposed by Ivanov and Guliev, 1988, 2002). It is supposed that one of major factors of the movement of sedimentary masses and formation of hydrocarbon deposits are phase transitions in sedimentary basin. At phase transitions there are abnormal changes of physical and chemical parameters of rocks. Abnormal (high and negative) pressure takes place. This process is called as excitation of the underground environment with periodicity from several tens to several hundreds, or thousand years. The relationship between mud volcanism and the generation of hydrocarbons, particularly methane, is considered to be a critical factor in mud volcano formation. At high flow rates the gas and sediment develops into a pseudo-liquid state and as flow increases the mass reaches the "so-called hover velocity" where mass transport begins. The mass of fluid moves as a quasi-uniform viscous mass through the sediment pile in a piston like manner until expelled from the surface as a "catastrophic eruption

  19. Tree Formation Using Coordinate Method

    Directory of Open Access Journals (Sweden)

    Monika Choudhary

    2015-06-01

    Full Text Available In this paper we are introducing a new method of tree formation, we propose a coordinate based method by which we can store and access tree structures. As we know in NLP, parsing is the most important module. The output of this module is generally parsed trees. Currently, TAG (Tree Adjoining Grammar is widely used grammar due to its linguistic and formal nature. It is simply tree generating system. The unit structure used in TAG is structured trees. So we used our new method to store trees where we worked on English to Hindi language. We worked on different sentences from English to Hindi, our method is the easiest way to manipulate tree. We have implemented within small corpus and for finite number of structures and further can be extended in future.

  20. Biofilm formation in Acinetobacter baumannii.

    Science.gov (United States)

    Longo, Francesca; Vuotto, Claudia; Donelli, Gianfranco

    2014-04-01

    Acinetobacter baumannii has received much attention in recent years because of its increasing involvement in a number of severe infections and outbreaks occurring in clinical settings, and presumably related to its ability to survive and persist in hospital environments. The treatment of infections caused by A. baumannii nosocomial strains has become increasingly problematic, due to their intrinsic and/or acquired resistance to multiple classes of antibiotics. Furthermore, the demonstrated ability of nosocomial strains to grow as biofilm is believed to play a significant role in their persistence and antibiotic resistance. This review summarises current knowledge on A. baumannii biofilm formation and its clinical significance, as well as the related genetic determinants and the regulation of this process.

  1. Pressure induced polymerization of Formates

    Science.gov (United States)

    Tschauner, Oliver

    2004-03-01

    The discovery of pressure induced polymerization of CO2 inspired us to search for C-O based chain structures forming at high pressure. We used salts of carboxylic acids as starting materials and exposed them to pressures between 10 and 30 GPa. Upon heating to temperatures above 1800 K we observed deprotonation and significant changes in the Raman shifts of C-O streching modes. Structure analysis based on powder diffraction patterns collected at sector 16 of the APS showed formation of extended C-O chain structures with the cations of the salts residing in the interchain spaces. These new high pressure polymers are interesting by their mechanical strength and provide basic molecular patterns of organic metallic conductors.

  2. Crystal formation in furunculosis agar

    Science.gov (United States)

    Bullock, G.L.; Ross, A.J.

    1964-01-01

    SINCE ITS INTRODUCTION SOME MONTHS AGO, FURUNCULOSIS AGAR has been employed in the diagnosis of suspect furunculosis and also as a general purpose medium. During our work with this medium we have noticed discrete "colonies," of crystalline material, which very closely resemble microbial colonies. These crystal colonies are compact and appear on both the surface and subsurface; they occur in inoculated slants and plates incubated for long periods (2 to 3 weeks), as well as in uninoculated stored medium. As the crystal colonies could be confusing to workers using this medium, we decided to attempt to identify them and also to determine whether storage conditions and different lots of medium affect crystal formation.

  3. Formation of $\\phi$ mesic nuclei

    CERN Document Server

    Yamagata-Sekihara, J; Vacas, M J Vicente; Hirenzaki, S

    2010-01-01

    We study the structure and formation of the $\\phi$ mesic nuclei to investigate the in-medium modification of the $\\phi$-meson spectral function at finite density. We consider (${\\bar p},\\phi$), ($\\gamma,p$) and ($\\pi^-,n$) reactions to produce a $\\phi$-meson inside the nucleus and evaluate the effects of its medium modifications to the reaction cross sections. We also estimate the consequences of the uncertainties of the ${\\bar K}$ selfenergy in medium to the $\\phi$-nucleus interaction. We find that it may be possible to see a peak structure in the reaction spectra for the strong attractive potential cases. On the other hand, for strong absorptive interaction cases with relatively weak attractions, it is very difficult to observe clear peaks and we may need to know the spectrum shape in a wide energy region to deduce the properties of $\\phi$.

  4. Formation of helical ion chains

    CERN Document Server

    Nigmatullin, Ramil; De Chiara, Gabriele; Morigi, Giovanna; Plenio, Martin B; Retzker, Alex

    2011-01-01

    We study the nonequilibrium dynamics of the linear to zigzag structural phase transition exhibited by an ion chain confined in a trap with periodic boundary conditions. The transition is driven by reducing the transverse confinement at a finite quench rate, which can be accurately controlled. This results in the formation of zigzag domains oriented along different transverse planes. The twists between different domains can be stabilized by the topology of the trap and under laser cooling the system relaxes to a helical chain with possibly nonzero winding number. Molecular dynamics simulations are used to obtain a large sample of possible trajectories for different quench rates. The scaling of the average winding number with different quench rates is compared to the prediction of the Kibble-Zurek theory, and a good quantitative agreement is found.

  5. Energy conditions and galaxy formation

    CERN Document Server

    Visser, M

    1997-01-01

    This note summarizes a model-independent analysis of the age of the universe problem that trades off precision in favour of robustness: The energy conditions of Einstein gravity are designed to extract as much information as possible from classical general relativity without specifying a particular equation of state. This is particularly useful in a cosmological setting, where the equation of state for the cosmological fluid is extremely uncertain. The strong energy condition (SEC) provides a simple and robust bound on the behaviour of the look-back time as a function of red-shift. Observation suggests that the SEC may be violated sometime between the epoch of galaxy formation and the present.

  6. On the formation of granulites

    Science.gov (United States)

    Bohlen, S.R.

    1991-01-01

    The tectonic settings for the formation and evolution of regional granulite terranes and the lowermost continental crust can be deduced from pressure-temperature-time (P-T-time) paths and constrained by petrological and geophysical considerations. P-T conditions deduced for regional granulites require transient, average geothermal gradients of greater than 35??C km-1, implying minimum heat flow in excess of 100 mW m-2. Such high heat flow is probably caused by magmatic heating. Tectonic settings wherein such conditions are found include convergent plate margins, continental rifts, hot spots and at the margins of large, deep-seated batholiths. Cooling paths can be constrained by solid-solid and devolatilization equilibria and geophysical modelling. -from Author

  7. New Office Software course format

    CERN Multimedia

    HR Department

    2010-01-01

    Always interested to anticipate your training needs, the Technical Training service is pleased to propose two new Office Software course formats : “Focus on... ” : On a monthly basis we will propose a theme such as “Sharepoint Collaboration Workspace” or “Word 2007” or “charts” etc. You will have to send us in advance your questions regarding the proposed topic and register for the course through our Training Catalogue. During the session, our trainer will answer all the questions received and participants will increase their knowledge thanks to the solutions discussed for everyone. The course will last two hours, from 09h00 to 11h00 - with open questions on the proposed topic at the end. “Office software Individual coaching”: If one or several particular topics cause you sleepless nights, you can get the help of our trainer who will come to your workplace for a multiple of 1-hour slots . All fields ...

  8. Magnetic Fields and Star Formation

    CERN Document Server

    Van Loo, S; Falle, S A E G

    2012-01-01

    Research performed in the 1950s and 1960s by Leon Mestel on the roles of magnetic fields in star formation established the framework within which he and other key figures have conducted subsequent investigations on the subject. This short tribute to Leon contains a brief summary of some, but not all, of his ground breaking contributions in the area. It also mentions of some of the relevant problems that have received attention in the last few years. The coverage is not comprehensive, and the authors have drawn on their own results more and touched more briefly on those of others than they would in a normal review. Theirs is a personal contribution to the issue honouring Leon, one of the truly great gentlemen, wits, and most insightful of astrophysicists.

  9. Engram Formation in Psychiatric Disorders

    Directory of Open Access Journals (Sweden)

    Peter J. Gebicke-Haerter

    2014-05-01

    Full Text Available Environmental factors substantially influence beginning and progression of mental illness, reinforcing or reducing the consequences of genetic vulnerability. Often initiated by early traumatic events, engrams or memories are formed that may give rise to a slow and subtle progression of psychiatric disorders. The large delay between beginning and time of onset (diagnosis may be explained by efficient compensatory mechanisms observed in brain metabolism that use optional pathways in highly redundant molecular interactions.To this end, research has to deal with mechanisms of learning and long-term memory formation, which involves a epigenetic changes, b altered neuronal activities and c changes in neuron-glia communication. On the epigenetic level, apparently DNA-methylations are more stable than histone modifications, although both closely interact. Neuronal activities basically deliver digital information, which clearly can serve as basis for memory formation (LTP. However, research in this respect has long time neglected the importance of glia. They are more actively involved in the control of neuronal activities than thought before. They can both reinforce and inhibit neuronal activities by transducing neuronal information from frequency-encoded to amplitude and frequency-modulated calcium wave patterns spreading in the glial syncytium by use of gap junctions. In this way, they serve integrative functions. In conclusion, we are dealing with two concepts of encoding information that mutually control each other and synergize: a digital (neuronal and a wave-like (glial computing, forming neuron-glia functional units with inbuilt feedback loops to maintain balance of excitation and inhibition. To better understand mental illness, we have to gain more insight into the dynamics of adverse environmental impact on those cellular and molecular systems. This report summarizes existing knowledge and draws some outline about further research in molecular

  10. Drill cuttings mount formation study

    Science.gov (United States)

    Teh, Su Yean; Koh, Hock Lye

    2014-07-01

    Oil, Gas and Energy sector has been identified as an essential driving force in the Malaysian Economic Transformation Programs (ETP). Recently confirmed discovery of many offshore oil and gas deposits in Malaysian waters has ignited new confidence in this sector. However, this has also spurred intense interest on safeguarding the health and environment of coastal waters in Malaysia from adverse impact resulting from offshore oil and gas production operation. Offshore discharge of spent drilling mud and rock cuttings is the least expensive and simplest option to dispose of large volumes of drilling wastes. But this onsite offshore disposal may have adverse environmental impacts on the water column and the seabed. It may also pose occupational health hazards to the workers living in the offshore platforms. It is therefore important to model the transport and deposition of drilling mud and rock cuttings in the sea to enable proper assessment of their adverse impacts on the environment and the workers. Further, accumulation of drill particles on the seabed may impede proper operation of pipelines on the seabed. In this paper, we present an in-house application model TUNA-PT developed to cater to local oil and gas industry needs to simulate the dispersion and mount formation of drill cuttings by offshore oil and gas exploration and production platforms. Using available data on Malaysian coastal waters, simulation analyses project a pile formation on the seabed with a maximum height of about 1 m and pile radius of around 30 to 50 m. Simulated pile heights are not sensitive to the heights of release of the cuttings as the sensitivity has been mitigated by the depth of water.

  11. Formation and Evolution of Protoatmospheres

    Science.gov (United States)

    Massol, H.; Hamano, K.; Tian, F.; Ikoma, M.; Abe, Y.; Chassefière, E.; Davaille, A.; Genda, H.; Güdel, M.; Hori, Y.; Leblanc, F.; Marcq, E.; Sarda, P.; Shematovich, V. I.; Stökl, A.; Lammer, H.

    2016-12-01

    The origin and evolution of planetary protoatmospheres in relation to the protoplanetary disk is discussed. The initial atmospheres of planets can mainly be related via two formation scenarios. If a protoplanetary core accretes mass and grows inside the gas disk, it can capture H2, He and other gases from the disk. When the gas of the disk evaporates, the core that is surrounded by the H2/He gas envelope is exposed to the high X-ray and extreme ultraviolet flux and stellar wind of the young host star. This period can be considered as the onset of atmospheric escape. It is shown that lower mass bodies accrete less gas and depending on the host stars radiation environment can therefore lose the gaseous envelope after tens or hundreds of million years. Massive cores may never get rid of their captured hydrogen envelopes and remain as sub-Neptunes, Neptunes or gas giants for their whole life time. Terrestrial planets which may have lost the captured gas envelope by thermal atmospheric escape, or which accreted after the protoplanetary nebula vanished will produce catastrophically outgassed steam atmospheres during the magma ocean solidification process. These steam atmospheres consist mainly of water and CO2 that was incorporated into the protoplanet during its accretion. Planets, which are formed in the habitable zone, solidify within several million years. In such cases the outgassed steam atmospheres cool fast, which leads to the condensation of water and the formation of liquid oceans. On the other hand, magma oceans are sustained for longer if planets form inside a critical distance, even if they outgassed a larger initial amount of water. In such cases the steam atmosphere could remain 100 million years or for even longer. Hydrodynamic atmospheric escape will then desiccate these planets during the slow solidification process.

  12. Formation and Evolution of Protoatmospheres

    Science.gov (United States)

    Massol, H.; Hamano, K.; Tian, F.; Ikoma, M.; Abe, Y.; Chassefière, E.; Davaille, A.; Genda, H.; Güdel, M.; Hori, Y.; Leblanc, F.; Marcq, E.; Sarda, P.; Shematovich, V. I.; Stökl, A.; Lammer, H.

    2016-09-01

    The origin and evolution of planetary protoatmospheres in relation to the protoplanetary disk is discussed. The initial atmospheres of planets can mainly be related via two formation scenarios. If a protoplanetary core accretes mass and grows inside the gas disk, it can capture H2, He and other gases from the disk. When the gas of the disk evaporates, the core that is surrounded by the H2/He gas envelope is exposed to the high X-ray and extreme ultraviolet flux and stellar wind of the young host star. This period can be considered as the onset of atmospheric escape. It is shown that lower mass bodies accrete less gas and depending on the host stars radiation environment can therefore lose the gaseous envelope after tens or hundreds of million years. Massive cores may never get rid of their captured hydrogen envelopes and remain as sub-Neptunes, Neptunes or gas giants for their whole life time. Terrestrial planets which may have lost the captured gas envelope by thermal atmospheric escape, or which accreted after the protoplanetary nebula vanished will produce catastrophically outgassed steam atmospheres during the magma ocean solidification process. These steam atmospheres consist mainly of water and CO2 that was incorporated into the protoplanet during its accretion. Planets, which are formed in the habitable zone, solidify within several million years. In such cases the outgassed steam atmospheres cool fast, which leads to the condensation of water and the formation of liquid oceans. On the other hand, magma oceans are sustained for longer if planets form inside a critical distance, even if they outgassed a larger initial amount of water. In such cases the steam atmosphere could remain 100 million years or for even longer. Hydrodynamic atmospheric escape will then desiccate these planets during the slow solidification process.

  13. MDF: Magnetic Particle Imaging Data Format

    CERN Document Server

    Knopp, Tobias; Bringout, Gael; Ahlborg, Mandy; Rahmer, Jürgen; Hofmann, Martin

    2016-01-01

    Magnetic particle imaging (MPI) is a tomographic method to determine the spatial distribution of magnetic nanoparticles. In this document a file format for the standardized storage of MPI data is introduced. The aim of the Magnetic Particle Imaging Data Format (MDF) is to provide a coherent way of exchanging MPI data acquired with different MPI scanners worldwide. The focus of the file format is on sequence parameters, raw measurement data, calibration data, and reconstruction data. The format is based on the hierarchical document format (HDF) in version 5 (HDF5).

  14. Format SPARQL Query Results into HTML Report

    Directory of Open Access Journals (Sweden)

    Dr Sunitha Abburu

    2013-07-01

    Full Text Available SPARQL is one of the powerful query language for querying semantic data. It is recognized by the W3C as a query language for RDF. As an efficient query language for RDF, it has defined several query result formats such as CSV, TSV and XML etc. These formats are not attractive, understandable and readable. The results need to be converted in an appropriate format so that user can easily understand. The above formats require additional transformations or tool support to represent the query result in user readable format. The main aim of this paper is to propose a method to build HTML report dynamically for SPARQL query results. This enables SPARQL query result display, in HTML report format easily, in an attractive understandable format without the support of any additional or external tools or transformation.

  15. Control of star formation by supersonic turbulence

    CERN Document Server

    MacLow, M M; Low, Mordecai-Mark Mac; Klessen, Ralf S.

    2004-01-01

    Understanding the formation of stars in galaxies is central to much of modern astrophysics. For several decades it has been thought that stellar birth is primarily controlled by the interplay between gravity and magnetostatic support, modulated by ambipolar diffusion. Recently, however, both observational and numerical work has begun to suggest that support by supersonic turbulence rather than magnetic fields controls star formation. In this review we outline a new theory of star formation relying on the control by turbulence. We demonstrate that although supersonic turbulence can provide global support, it nevertheless produces density enhancements that allow local collapse. Inefficient, isolated star formation is a hallmark of turbulent support, while efficient, clustered star formation occurs in its absence. The consequences of this theory are then explored for both local star formation and galactic scale star formation. (Abstract abbreviated)

  16. Improving the Formatting Tools of CDS Invenio

    CERN Document Server

    Caffaro, J; Pu Faltings, Pearl

    2006-01-01

    CDS Invenio is the web-based integrated digital library system developed at CERN. It is a strategical tool that supports the archival and open dissemination of documents produced by CERN researchers. This paper reports on my Master’s thesis work done on BibFormat, a module in CDS Invenio, which formats documents metadata. The goal of this project was to implement a completely new formatting module for CDS Invenio. In this report a strong emphasis is put on the user-centered design of the new BibFormat. The bibliographic formatting process and its requirements are discussed. The task analysis and its resulting interaction model are detailed. The document also shows the implemented user interface of BibFormat and gives the results of the user evaluation of this interface. Finally the results of a small usability study of the formats included in CDS Invenio are discussed.

  17. Fibril formation from pea protein and subsequent gel formation.

    Science.gov (United States)

    Munialo, Claire Darizu; Martin, Anneke H; van der Linden, Erik; de Jongh, Harmen H J

    2014-03-19

    The objective of this study was to characterize fibrillar aggregates made using pea proteins, to assemble formed fibrils into protein-based gels, and to study the rheological behavior of these gels. Micrometer-long fibrillar aggregates were observed after pea protein solutions had been heated for 20 h at pH 2.0. Following heating of pea proteins, it was observed that all of the proteins were hydrolyzed into peptides and that 50% of these peptides were assembled into fibrils. Changes on a structural level in pea proteins were studied using circular dichroism, transmission electron microscopy, and particle size analysis. During the fibril assembly process, an increase in aggregate size was observed, which coincided with an increase in thioflavin T binding, indicating the presence of β-sheet aggregates. Fibrils made using pea proteins were more branched and curly. Gel formation of preformed fibrils was induced by slow acidification from pH 7.0 to a final pH of around pH 5.0. The ability of pea protein-based fibrillar gels to fracture during an amplitude sweep was comparable to those of soy protein and whey protein-based fibrillar gels, although gels prepared from fibrils made using pea protein and soy protein were weaker than those of whey protein. The findings show that fibrils can be prepared from pea protein, which can be incorporated into protein-based fibrillar gels.

  18. Magnetic Assisted Colloidal Pattern Formation

    Science.gov (United States)

    Yang, Ye

    Pattern formation is a mysterious phenomenon occurring at all scales in nature. The beauty of the resulting structures and myriad of resulting properties occurring in naturally forming patterns have attracted great interest from scientists and engineers. One of the most convenient experimental models for studying pattern formation are colloidal particle suspensions, which can be used both to explore condensed matter phenomena and as a powerful fabrication technique for forming advanced materials. In my thesis, I have focused on the study of colloidal patterns, which can be conveniently tracked in an optical microscope yet can also be thermally equilibrated on experimentally relevant time scales, allowing for ground states and transitions between them to be studied with optical tracking algorithms. In particular, I have focused on systems that spontaneously organize due to particle-surface and particle-particle interactions, paying close attention to systems that can be dynamically adjusted with an externally applied magnetic or acoustic field. In the early stages of my doctoral studies, I developed a magnetic field manipulation technique to quantify the adhesion force between particles and surfaces. This manipulation technique is based on the magnetic dipolar interactions between colloidal particles and their "image dipoles" that appear within planar substrate. Since the particles interact with their own images, this system enables massively parallel surface force measurements (>100 measurements) in a single experiment, and allows statistical properties of particle-surface adhesion energies to be extracted as a function of loading rate. With this approach, I was able to probe sub-picoNewton surface interactions between colloidal particles and several substrates at the lowest force loading rates ever achieved. In the later stages of my doctoral studies, I focused on studying patterns formed from particle-particle interaction, which serve as an experimental model of

  19. Vol en formation sans formation: contrôle et planification pour le vol en formation des avions sans pilote

    OpenAIRE

    2008-01-01

    This thesis presents the study and the implementation of a system that manages autonomously the configuration of a formation of unmanned aerial vehicles, or UAVs. The purposes are to improve the safety and the efficiency of a group of military UAVs, and to fill the gap between mission planning layers and formation control layers. Formation flight is particularly suited for military applications in hostile environments, that require synchronizations on target arrivals or mutual support for jam...

  20. Star Formation in Tadpole Galaxies

    CERN Document Server

    Munoz-Tunon, Casiana; Elmegreen, Debra M; Elmegreen, Bruce G

    2014-01-01

    Tadpole Galaxies look like a star forming head with a tail structure to the side. They are also named cometaries. In a series of recent works we have discovered a number of issues that lead us to consider them extremely interesting targets. First, from images, they are disks with a lopsided starburst. This result is firmly established with long slit spectroscopy in a nearby representative sample. They rotate with the head following the rotation pattern but displaced from the rotation center. Moreover, in a search for extremely metal poor (XMP) galaxies, we identified tadpoles as the dominant shapes in the sample- nearly 80% of the local XMP galaxies have a tadpole morphology. In addition, the spatially resolved analysis of the metallicity shows the remarkable result that there is a metallicity drop right at the position of the head. This is contrary to what intuition would say and difficult to explain if star formation has happened from gas processed in the disk. The result could however be understood if the ...