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Sample records for vesicular stomatitis virus-indiana

  1. Display of disparate transcription phenotype by the phosphorylation negative P protein mutants of vesicular stomatitis virus, Indiana serotype, expressed in E. coli and eucaryotic cells.

    Science.gov (United States)

    Mathur, M; Das, T; Chen, J L; Chattopadhyay, D; Banerjee, A K

    1997-01-01

    The phosphoprotein (P) of vesicular stomatitis virus (VSV) is a subunit of the RNA polymerase (L) that transcribes the negative strand genome RNA into mRNAs both in vitro and in vivo. We have recently shown that the P protein of VSV, New Jersey serotype (PNJ), expressed in E. coli, is biologically inactive unless phosphorylated at specific serine residues by cellular casein kinase II (CKII). In the present work, we are studying the role of phosphorylation in the activation of the P protein of Indiana serotype (PIND), which is highly nonhomologous in amino acid sequence yet structurally similar to its New Jersey counterpart. Despite the fact that E. coli-expressed PIND required phosphorylation by CKII for activation, the phosphorylation negative P protein mutants generated by altering the phosphate acceptors S and T to alanine, surprisingly, showed transcription activity similar to wild-type in vitro. Alteration of S and T residues to phenylalanine, similarly, supported substantial transcription activity (approx. 60% of wild-type), whereas substitution with arginine residue abrogated transcription (approx. 5% of wild-type). In contrast, the same mutants, when expressed in eucaryotic cells, exhibited greatly reduced transcription activity in vitro. This disparate display of transcription phenotype by the PIND mutants expressed in bacteria and eucaryotic cells suggests that these mutants are unique in assuming different secondary structure or conformation when synthesized in two different cellular milieu. The findings that, unless phosphorylated by CKII, the bacterially expressed unphosphorylated (P0) form of PIND, as well as the phosphorylation negative mutants expressed in eucaryotic cells, demonstrates transcription negative phenotype indicate that, like PNJ, phosphorylation of PIND is essential for its activity.

  2. Vesicular stomatitis virus (indiana 2 serotype as experimental model to study acute encephalitis – morphological features Vírus da estomatite vesicular (sorotipo indiana 2 como modelo experimental para o estudo de encefalite aguda – aspectos morfológicos

    Directory of Open Access Journals (Sweden)

    Florêncio Figueiredo Cavalcanti Neto

    2003-10-01

    Full Text Available The Vesicular Stomatitis Virus (VSV is a Vesiculovirus of the Rhabdoviridae family that infects mammals and causes vesicular lesions similar to those of foot-and-mouth disease. VSV experimental encephalitis can be induced in rodents and the symptoms are similar to those observed in rabies. However, the lesions observed in the animals´ encephalon are different. Inclusion bodies are not observed. There is necrosis, particularly in the region of the olfactory bulb, and, in some cases, ventriculitis. It was observed that the time pattern of VSV dissemination and the morphological aspects of the lesions are similar to those described in literature. The virus seems to be disseminated through the brain ventricles, being multiplied in the ependyma cells and in the neurons, besides using retrograde and anterograde transport. It was noticed that, due to the facility of virus manipulation, this experimental model has been used in innumerable research studies in several fields. If, on the one hand there are plenty of reports on the infection pathogenesis, on the other hand there are many gaps involving, for instance, aspects about virus transmission, recovery of infected animals and participation of glial cells in the acute as well as in the recovery phases.   O vírus da estomatite vesicular (VEV é um Vesiculovírus da família Rhabdoviridae que infecta mamíferos e causa lesões vesiculares semelhantes às observadas na febre aftosa. A encefalite experimental pode ser induzida em roedores e os sintomas são semelhantes aos observados na raiva; entretanto, as lesões observadas no encéfalo dos animais são diferentes. Corpúsculos de inclusão não são observados, há necrose especialmente da região do bulbo olfatório e em alguns casos, ventriculite. Observamos que o padrão temporal de disseminação do VEV e os aspectos morfológicos das lesões são similares aos descritos na literatura. O vírus parece se disseminar através dos ventr

  3. Vesicular stomatitis forecasting based on Google Trends

    Science.gov (United States)

    Lu, Yi; Zhou, GuangYa; Chen, Qin

    2018-01-01

    Background Vesicular stomatitis (VS) is an important viral disease of livestock. The main feature of VS is irregular blisters that occur on the lips, tongue, oral mucosa, hoof crown and nipple. Humans can also be infected with vesicular stomatitis and develop meningitis. This study analyses 2014 American VS outbreaks in order to accurately predict vesicular stomatitis outbreak trends. Methods American VS outbreaks data were collected from OIE. The data for VS keywords were obtained by inputting 24 disease-related keywords into Google Trends. After calculating the Pearson and Spearman correlation coefficients, it was found that there was a relationship between outbreaks and keywords derived from Google Trends. Finally, the predicted model was constructed based on qualitative classification and quantitative regression. Results For the regression model, the Pearson correlation coefficients between the predicted outbreaks and actual outbreaks are 0.953 and 0.948, respectively. For the qualitative classification model, we constructed five classification predictive models and chose the best classification predictive model as the result. The results showed, SN (sensitivity), SP (specificity) and ACC (prediction accuracy) values of the best classification predictive model are 78.52%,72.5% and 77.14%, respectively. Conclusion This study applied Google search data to construct a qualitative classification model and a quantitative regression model. The results show that the method is effective and that these two models obtain more accurate forecast. PMID:29385198

  4. Understanding and altering cell tropism of vesicular stomatitis virus

    OpenAIRE

    Hastie, Eric; Cataldi, Marcela; Marriott, Ian; Valery Z Grdzelishvili

    2013-01-01

    Vesicular stomatitis virus (VSV) is a prototypic nonsegmented negative-strand RNA virus. VSV’s broad cell tropism makes it a popular model virus for many basic research applications. In addition, a lack of preexisting human immunity against VSV, inherent oncotropism and other features make VSV a widely used platform for vaccine and oncolytic vectors. However, VSV’s neurotropism that can result in viral encephalitis in experimental animals needs to be addressed for the use of the virus as a sa...

  5. Lassa-vesicular stomatitis chimeric virus safely destroys brain tumors.

    Science.gov (United States)

    Wollmann, Guido; Drokhlyansky, Eugene; Davis, John N; Cepko, Connie; van den Pol, Anthony N

    2015-07-01

    High-grade tumors in the brain are among the deadliest of cancers. Here, we took a promising oncolytic virus, vesicular stomatitis virus (VSV), and tested the hypothesis that the neurotoxicity associated with the virus could be eliminated without blocking its oncolytic potential in the brain by replacing the neurotropic VSV glycoprotein with the glycoprotein from one of five different viruses, including Ebola virus, Marburg virus, lymphocytic choriomeningitis virus (LCMV), rabies virus, and Lassa virus. Based on in vitro infections of normal and tumor cells, we selected two viruses to test in vivo. Wild-type VSV was lethal when injected directly into the brain. In contrast, a novel chimeric virus (VSV-LASV-GPC) containing genes from both the Lassa virus glycoprotein precursor (GPC) and VSV showed no adverse actions within or outside the brain and targeted and completely destroyed brain cancer, including high-grade glioblastoma and melanoma, even in metastatic cancer models. When mice had two brain tumors, intratumoral VSV-LASV-GPC injection in one tumor (glioma or melanoma) led to complete tumor destruction; importantly, the virus moved contralaterally within the brain to selectively infect the second noninjected tumor. A chimeric virus combining VSV genes with the gene coding for the Ebola virus glycoprotein was safe in the brain and also selectively targeted brain tumors but was substantially less effective in destroying brain tumors and prolonging survival of tumor-bearing mice. A tropism for multiple cancer types combined with an exquisite tumor specificity opens a new door to widespread application of VSV-LASV-GPC as a safe and efficacious oncolytic chimeric virus within the brain. Many viruses have been tested for their ability to target and kill cancer cells. Vesicular stomatitis virus (VSV) has shown substantial promise, but a key problem is that if it enters the brain, it can generate adverse neurologic consequences, including death. We tested a series of

  6. Understanding and altering cell tropism of vesicular stomatitis virus

    Science.gov (United States)

    Hastie, Eric; Cataldi, Marcela; Marriott, Ian; Grdzelishvili, Valery Z.

    2013-01-01

    Vesicular stomatitis virus (VSV) is a prototypic nonsegmented negative-strand RNA virus. VSV’s broad cell tropism makes it a popular model virus for many basic research applications. In addition, a lack of preexisting human immunity against VSV, inherent oncotropism and other features make VSV a widely used platform for vaccine and oncolytic vectors. However, VSV’s neurotropism that can result in viral encephalitis in experimental animals needs to be addressed for the use of the virus as a safe vector. Therefore, it is very important to understand the determinants of VSV tropism and develop strategies to alter it. VSV glycoprotein (G) and matrix (M) protein play major roles in its cell tropism. VSV G protein is responsible for VSV broad cell tropism and is often used for pseudotyping other viruses. VSV M affects cell tropism via evasion of antiviral responses, and M mutants can be used to limit cell tropism to cell types defective in interferon signaling. In addition, other VSV proteins and host proteins may function as determinants of VSV cell tropism. Various approaches have been successfully used to alter VSV tropism to benefit basic research and clinically relevant applications. PMID:23796410

  7. Asymmetric packaging of polymerases within vesicular stomatitis virus

    Energy Technology Data Exchange (ETDEWEB)

    Hodges, Jeffery; Tang, Xiaolin; Landesman, Michael B. [Dept. of Physics and Astronomy, University of Utah (United States); Center for Cell and Genome Science, University of Utah (United States); Ruedas, John B. [Dept. of Biology, San Diego State University (United States); Ghimire, Anil [Dept. of Physics and Astronomy, University of Utah (United States); Gudheti, Manasa V. [Vutara, Inc., Salt Lake City, UT (United States); Dept. of Biology, University of Utah (United States); Perrault, Jacques [Dept. of Biology, San Diego State University (United States); Jorgensen, Erik M. [Howard Hughes Medical Institute (United States); Dept. of Biology, University of Utah (United States); Gerton, Jordan M. [Dept. of Physics and Astronomy, University of Utah (United States); Dept. of Bioengineering, University of Utah (United States); Saffarian, Saveez, E-mail: saffarian@physics.utah.edu [Dept. of Physics and Astronomy, University of Utah (United States); Center for Cell and Genome Science, University of Utah (United States); Dept. of Biology, University of Utah (United States)

    2013-10-18

    Highlights: •The VSV polymerases (L proteins) are localized to the blunt end of the virus. •The VSV phosphoproteins (P proteins) are localized to the blunt end of the virus. •Each VSV virion packages a variable number of P and L proteins. -- Abstract: Vesicular stomatitis virus (VSV) is a prototypic negative sense single-stranded RNA virus. The bullet-shape appearance of the virion results from tightly wound helical turns of the nucleoprotein encapsidated RNA template (N-RNA) around a central cavity. Transcription and replication require polymerase complexes, which include a catalytic subunit L and a template-binding subunit P. L and P are inferred to be in the cavity, however lacking direct observation, their exact position has remained unclear. Using super-resolution fluorescence imaging and atomic force microscopy (AFM) on single VSV virions, we show that L and P are packaged asymmetrically towards the blunt end of the virus. The number of L and P proteins varies between individual virions and they occupy 57 ± 12 nm of the 150 nm central cavity of the virus. Our finding positions the polymerases at the opposite end of the genome with respect to the only transcriptional promoter.

  8. Understanding and altering cell tropism of vesicular stomatitis virus.

    Science.gov (United States)

    Hastie, Eric; Cataldi, Marcela; Marriott, Ian; Grdzelishvili, Valery Z

    2013-09-01

    Vesicular stomatitis virus (VSV) is a prototypic nonsegmented negative-strand RNA virus. VSV's broad cell tropism makes it a popular model virus for many basic research applications. In addition, a lack of preexisting human immunity against VSV, inherent oncotropism and other features make VSV a widely used platform for vaccine and oncolytic vectors. However, VSV's neurotropism that can result in viral encephalitis in experimental animals needs to be addressed for the use of the virus as a safe vector. Therefore, it is very important to understand the determinants of VSV tropism and develop strategies to alter it. VSV glycoprotein (G) and matrix (M) protein play major roles in its cell tropism. VSV G protein is responsible for VSV broad cell tropism and is often used for pseudotyping other viruses. VSV M affects cell tropism via evasion of antiviral responses, and M mutants can be used to limit cell tropism to cell types defective in interferon signaling. In addition, other VSV proteins and host proteins may function as determinants of VSV cell tropism. Various approaches have been successfully used to alter VSV tropism to benefit basic research and clinically relevant applications. Copyright © 2013 Elsevier B.V. All rights reserved.

  9. Molecular architecture of the vesicular stomatitis virus RNA polymerase.

    Science.gov (United States)

    Rahmeh, Amal A; Schenk, Andreas D; Danek, Eric I; Kranzusch, Philip J; Liang, Bo; Walz, Thomas; Whelan, Sean P J

    2010-11-16

    Nonsegmented negative-strand (NNS) RNA viruses initiate infection by delivering into the host cell a highly specialized RNA synthesis machine comprising the genomic RNA completely encapsidated by the viral nucleocapsid protein and associated with the viral polymerase. The catalytic core of this protein-RNA complex is a 250-kDa multifunctional large (L) polymerase protein that contains enzymatic activities for nucleotide polymerization as well as for each step of mRNA cap formation. Working with vesicular stomatitis virus (VSV), a prototype of NNS RNA viruses, we used negative stain electron microscopy (EM) to obtain a molecular view of L, alone and in complex with the viral phosphoprotein (P) cofactor. EM analysis, combined with proteolytic digestion and deletion mapping, revealed the organization of L into a ring domain containing the RNA polymerase and an appendage of three globular domains containing the cap-forming activities. The capping enzyme maps to a globular domain, which is juxtaposed to the ring, and the cap methyltransferase maps to a more distal and flexibly connected globule. Upon P binding, L undergoes a significant rearrangement that may reflect an optimal positioning of its functional domains for transcription. The structural map of L provides new insights into the interrelationship of its various domains, and their rearrangement on P binding that is likely important for RNA synthesis. Because the arrangement of conserved regions involved in catalysis is homologous, the structural insights obtained for VSV L likely extend to all NNS RNA viruses.

  10. Redistributive properties of the vesicular stomatitis virus polymerase

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    Helfman, W.B.; Perrault, J. (San Diego State Univ., CA (USA))

    1989-08-01

    The template for transcription of the vesicular stomatitis virus (VSV) genome consists of a negative-strand RNA (approximately 11 kb) tightly associated with approximately 1250 copies of the nucleocapsid or N protein (N-RNA template). The interaction between the virion-associated polymerase and this template was probed with a novel assay using purified N-RNA complexes added to detergent-disrupted uv-irradiated standard virions or unirradiated defective interfering (DI) particles. In contrast to the well-known stability of assembled cellular transcription complexes, the VSV polymerase copied exogenously added templates efficiently and yielded products indistinguishable from control virus transcription. Addition of uv-irradiated N-RNA templates to unirradiated virus effectively competed for transcription of endogenous template indicating that most or all of the polymerase can freely redistribute. Furthermore preincubation of virus and added templates at high ionic strength to solubilize L and NS polymerase proteins did not release additional active enzyme for redistribution. Pretranscription of virus also had little or no effect on redistributed activity indicating that polymerase complexes are capable of multiple rounds of synthesis beginning at the 3' end promoter. Unexpectedly, titration with saturating amounts of added N-RNA showed that active polymerase complexes are only in slight excess relative to template in standard or DI particles despite the large surplus of packaged L and NS polypeptides. Moreover, added standard virus templates competed equally well for the redistributing polymerase from DI particles or standard virus indicating no significant polymerase-binding preference for interfering templates. These findings bear important implications regarding mechanisms of VSV transcription and replication.

  11. Inhibition of cellular DNA synthesis by vesicular stomatitis virus

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    McGowan, J.J.; Wagner, R.R.

    1981-04-01

    DNA synthesis in mouse myeloma (MPC-11) cells and L cells was rapidly and progressively inhibited by infection with vesicular stomatitis virus (VSV). No significant difference in cellular DNA synthesis inhibition was noted between synchronized and unsynchronized cells, nor did synchronized cells vary in their susceptibility to VSV infection after release from successive thymidine and hydroxyurea blocks. Cellular RNA synthesis was inhibited to about the same extent as DNA synthesis, but cellular protein synthesis was less affected by VSV at the same multiplicity of infection. The effect of VSV on cellular DNA synthesis could not be attributed to degradation of existing DNA or to decreased uptake of deoxynucleoside triphosphates, nor were DNA polymerase and thymidine kinase activities significantly different in VSV-infected and uninfected cell extracts. Analysis by alkaline sucrose gradients of DNA in pulse-labeled uninfected and VSV-infected cells indicated that VSV infection did not appear to influence DNA chain elongation. Cellular DNA synthesis was not significantly inhibited by infection with the VSV polymerase mutant tsG114(I) at the restrictive temperature or by infection with defective-interfering VSV DI-011 (5' end of the genome), but DI-HR-LT (3' end of genome) exhibited initially rapid but not prolonged inhibition of MPC-11 cell DNA synthesis. DNA synthesis inhibitory activity of wild-type VSV was only slowly and partially inactivated by very large doses of UV irradiation. These data suggest that, as in the effect of VSV on cellular RNA synthesis inhibition of cellular DNA synthesis by VSV requires transcription of a small segment of the viral genome.

  12. Vesicular stomatitis virus-based vaccines against Lassa and Ebola viruses.

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    Marzi, Andrea; Feldmann, Friederike; Geisbert, Thomas W; Feldmann, Heinz; Safronetz, David

    2015-02-01

    We demonstrated that previous vaccination with a vesicular stomatitis virus (VSV)-based Lassa virus vaccine does not alter protective efficacy of subsequent vaccination with a VSV-based Ebola virus vaccine. These findings demonstrate the utility of VSV-based vaccines against divergent viral pathogens, even when preexisting immunity to the vaccine vector is present.

  13. TNF-mediated survival of CD169(+) cells promotes immune activation during vesicular stomatitis virus infection

    DEFF Research Database (Denmark)

    Shinde, Prashant V; Xu, Haifeng C; Maney, Sathish Kumar

    2017-01-01

    Innate immune activation is essential to mount an effective antiviral response and to prime adaptive immunity. Although a crucial role of CD169(+) cells during vesicular stomatitis virus (VSV) infections is increasingly recognized, factors regulating CD169(+) cells during viral infections remain ...

  14. Neurovirulence properties of recombinant vesicular stomatitis virus vectors in non-human primates

    OpenAIRE

    Johnson, J. Erik; Nasar, Farooq; Coleman, John W.; Price, Roger E; Javadian, Ali; Draper, Kenneth; Lee, Margaret; Reilly, Patricia A.; Clarke, David K.; Hendry, R. Michael; Udem, Stephen A.

    2007-01-01

    Although vesicular stomatitis virus (VSV) neurovirulence and pathogenicity in rodents have been well studied, little is known about VSV pathogenicity in non-human primates. To address this question, we measured VSV viremia, shedding, and neurovirulence in macaques. Following intranasal inoculation, macaques shed minimal recombinant VSV (rVSV) in nasal washes for one day post-inoculation; viremia was not detected. Following intranasal inoculation of macaques, wild type (wt) VSV, rVSV, and two ...

  15. Protective efficacy of a recombinant Newcastle disease virus expressing glycoprotein of vesicular stomatitis virus in mice

    OpenAIRE

    Zhang, Minmin; Ge, Jinying; Li, Xiaofang; Chen, Weiye; Wang, Xijun; Wen, Zhiyuan; Bu, Zhigao

    2016-01-01

    Background Vesicular stomatitis virus (VSV) causes severe losses to the animal husbandry industry. In this study, a recombinant Newcastle disease virus (NDV) expressing the glycoprotein (G) of VSV (rL-VSV-G) was constructed and its pathogenicity and immune protective efficacy in mouse were evaluated. Results In pathogenicity evaluation test, the analysis of the viral distribution in mouse organs and body weight change showed that rL-VSV-G was safe in mice. In immune protection assay, the reco...

  16. Pseudotype formation of murine leukemia virus with the G protein of vesicular stomatitis virus.

    OpenAIRE

    Emi, N; Friedmann, T; Yee, J K

    1991-01-01

    Mixed infection of a cell by vesicular stomatitis virus (VSV) and retroviruses results in the production of progeny virions bearing the genome of one virus encapsidated by the envelope proteins of the other. The mechanism for the phenomenon of pseudotype formation is not clear, although specific recognition of a viral envelope protein by the nucleocapsid of an unrelated virus is presumably involved. In this study, we used Moloney murine leukemia virus (MoMLV)-based retroviral vectors encoding...

  17. Production of vesicular stomatitis virus by antigen- or mitogen-stimulated lymphocytes and continuous lymphoblastoid lines

    Energy Technology Data Exchange (ETDEWEB)

    Nowakowski, M.; Feldman, J.D.; Kano, S.; Bloom, B.R.

    1973-04-01

    The purpose of this study is to explore at the ultrastructural level the nature of the cells engaged in the production of vesicular stomatitis virus (VSV) in different lymphoid cell populations, particularly after stimulation with several different agents. Specifically, we have examined (a) lymph node cells from guinea pigs with delayed hypersensitivity activated by specific antigen, (b) murine spleen cells activated by selective B cell and T cell mitogens, and (c) cells of human and murine continuous lymphoblastoid or lymphoma lines.

  18. Cytopathogenesis of Vesicular Stomatitis virus is regulated by the PSAP motif of M protein in a species-dependent manner

    Science.gov (United States)

    Vesicular stomatitis virus (VSV) is an important vector-borne pathogen of bovine and equine species, causing a reportable vesicular disease. The matrix (M) protein of VSV is multifunctional and plays a key role in cytopathogenesis, apoptosis, host protein shut-off, and virion assembly/budding. Our ...

  19. Radiation inactivation analysis of fusion and hemolysis by vesicular stomatitis virus

    Energy Technology Data Exchange (ETDEWEB)

    Bundo-Morita, K.; Gibson, S.; Lenard, J.

    1988-04-01

    Radiation inactivation analysis was used to determine the size of the functional unit responsible for fusion of vesicular stomatitis virus (VSV) with cardiolipin or phosphatidylcholine-phosphatidylethanolamine (1:1) liposomes, and for VSV-induced hemolysis. When radiation-insensitive background values were subtracted, the calculated functional units for all three activities were similar, ranging from 866 to 957 kDa, equivalent to about 15 G protein molecules. This is in striking contrast to results of similar studies with influenza and Sendai viruses, in which the functional unit corresponded in size to a single fusion protein monomer, and suggests that VSV fusion may occur by a different mechanism.

  20. Initiation and Direction of RNA Transcription by Vesicular Stomatitis Virus Virion Transcriptase

    Science.gov (United States)

    Roy, Polly; Bishop, D. H. L.

    1973-01-01

    The initiation of RNA transcription by the virion-bound RNA transcriptase of vesicular stomatitis virus has been examined. Multiple initiation sequences have been observed, two of which have been characterized (pppApCpGp... and pppGpCp...) suggestive of a transcription process which can start at different sites along the template RNA. By the use of sequential labeling techniques and exonucleases, it has been determined that there is a 5′ to 3′ direction of product RNA synthesis. PMID:4349490

  1. Vesicular Stomatitis Virus Infection Promotes Immune Evasion by Preventing NKG2D-Ligand Surface Expression

    DEFF Research Database (Denmark)

    Jensen, Helle; Andresen, Lars; Nielsen, Jens

    2011-01-01

    Vesicular stomatitis virus (VSV) has recently gained attention for its oncolytic ability in cancer treatment. Initially, we hypothesized that VSV infection could increase immune recognition of cancer cells through induction of the immune stimulatory NKG2D-ligands. Here we show that VSV infection......D-ligand expression at an early post-transcriptional level. Our results show that VSV possess an escape mechanism, which could affect the immune recognition of VSV infected cancer cells. This may also have implications for immune recognition of cancer cells after combined treatment with VSV...

  2. Oncolytic recombinant vesicular stomatitis virus (VSV) is nonpathogenic and non-transmissible in pigs, a natural host of VSV

    Science.gov (United States)

    Vesicular stomatitis virus (VSV) is a negative stranded RNA virus that naturally causes disease in agricultural livestock including horses, cattle and pigs. The two main identified VSV strains are the New Jersey (VSNJV) and Indiana (VSIV) strains. VSV is a rapidly replicating, potently immunogenic v...

  3. LDL receptor and its family members serve as the cellular receptors for vesicular stomatitis virus

    Science.gov (United States)

    Finkelshtein, Danit; Werman, Ariel; Novick, Daniela; Barak, Sara; Rubinstein, Menachem

    2013-01-01

    Vesicular stomatitis virus (VSV) exhibits a remarkably robust and pantropic infectivity, mediated by its coat protein, VSV-G. Using this property, recombinant forms of VSV and VSV-G-pseudotyped viral vectors are being developed for gene therapy, vaccination, and viral oncolysis and are extensively used for gene transduction in vivo and in vitro. The broad tropism of VSV suggests that it enters cells through a highly ubiquitous receptor, whose identity has so far remained elusive. Here we show that the LDL receptor (LDLR) serves as the major entry port of VSV and of VSV-G-pseudotyped lentiviral vectors in human and mouse cells, whereas other LDLR family members serve as alternative receptors. The widespread expression of LDLR family members accounts for the pantropism of VSV and for the broad applicability of VSV-G-pseudotyped viral vectors for gene transduction. PMID:23589850

  4. Impact of Vesicular Stomatitis Virus M Proteins on Different Cellular Functions.

    Directory of Open Access Journals (Sweden)

    Natalia Redondo

    Full Text Available Three different matrix (M proteins termed M1, M2 and M3 have been described in cells infected with vesicular stomatitis virus (VSV. Individual expression of VSV M proteins induces an evident cytopathic effect including cell rounding and detachment, in addition to a partial inhibition of cellular protein synthesis, likely mediated by an indirect mechanism. Analogous to viroporins, M1 promotes the budding of new virus particles; however, this process does not produce an increase in plasma membrane permeability. In contrast to M1, M2 and M3 neither interact with the cellular membrane nor promote the budding of double membrane vesicles at the cell surface. Nonetheless, all three species of M protein interfere with the transport of cellular mRNAs from the nucleus to the cytoplasm and also modulate the redistribution of the splicing factor. The present findings indicate that all three VSV M proteins share some activities that interfere with host cell functions.

  5. Spectrum of Text Information Content in the RNA Sequence of the Vesicular Stomatitis Virus

    Science.gov (United States)

    Filyukov, Alexander A.

    A new strategy to recognize patterns in the DNA sequences with functional significance is proposed. The strategy is based on the general definition of any individual organism as a Gibbsian ensemble of identical personal DNA molecules. This approach provides application of the methods of statistical thermodynamics of irreversible steady processes to genome informatics. The random processes theory and its Markov chains approximation lead in this approach directly to the definition of the generalized concept of evolution entropy and to the genuine measure of text information content in the sequences. Computer-assisted proofs of the existence of the nonequilibrium steady state conditions in genome molecule were obtained by investigation of the special type balance relations in the vesicular stomatitis virus (VSV) RNA sequence. The main maxima of the text information content were decoded and denominated. The established coding principles are connected with deviations from equilibrium conditions and from equipartition.

  6. Pseudotyping of vesicular stomatitis virus with the envelope glycoproteins of highly pathogenic avian influenza viruses.

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    Zimmer, Gert; Locher, Samira; Berger Rentsch, Marianne; Halbherr, Stefan J

    2014-08-01

    Pseudotype viruses are useful for studying the envelope proteins of harmful viruses. This work describes the pseudotyping of vesicular stomatitis virus (VSV) with the envelope glycoproteins of highly pathogenic avian influenza viruses. VSV lacking the homotypic glycoprotein (G) gene (VSVΔG) was used to express haemagglutinin (HA), neuraminidase (NA) or the combination of both. Propagation-competent pseudotype viruses were only obtained when HA and NA were expressed from the same vector genome. Pseudotype viruses containing HA from different H5 clades were neutralized specifically by immune sera directed against the corresponding clade. Fast and sensitive reading of test results was achieved by vector-mediated expression of GFP. Pseudotype viruses expressing a mutant VSV matrix protein showed restricted spread in IFN-competent cells. This pseudotype system will facilitate the detection of neutralizing antibodies against virulent influenza viruses, circumventing the need for high-level biosafety containment. © 2014 The Authors.

  7. Cutting Edge: Innate Immune Augmenting Vesicular Stomatitis Virus Expressing Zika Virus Proteins Confers Protective Immunity.

    Science.gov (United States)

    Betancourt, Dillon; de Queiroz, Nina M G P; Xia, Tianli; Ahn, Jeonghyun; Barber, Glen N

    2017-04-15

    Zika virus (ZIKV) has become a serious public health concern because of its link to brain damage in developing human fetuses. Recombinant vesicular stomatitis virus (rVSV) was shown to be a highly effective and safe vector for the delivery of foreign immunogens for vaccine purposes. In this study, we generated rVSVs (wild-type and attenuated VSV with mutated matrix protein [VSVm] versions) that express either the full length ZIKV envelope protein (ZENV) alone or include the ZENV precursor to the membrane protein upstream of the envelope protein, and our rVSV-ZIKV constructs showed efficient immunogenicity in murine models. We also demonstrated maternal protective immunity in challenged newborn mice born to female mice vaccinated with VSVm-ZENV containing the transmembrane domain. Our data indicate that rVSVm may be a suitable strategy for the design of effective vaccines against ZIKV. Copyright © 2017 by The American Association of Immunologists, Inc.

  8. Pathogenesis of experimental vesicular stomatitis virus (New Jersey serotype) infection in the deer mouse (Peromyscus maniculatus).

    Science.gov (United States)

    Cornish, T E; Stallknecht, D E; Brown, C C; Seal, B S; Howerth, E W

    2001-07-01

    The pathogenesis of vesicular stomatitis virus (VSV) infection has not been investigated previously in native New World rodents that may have a role in the epidemiology of the disease. In the present study, 45 juvenile and 80 adult deer mice (Peromyscus maniculatus) were inoculated intranasally with VSV New Jersey serotype (VSV-NJ) and examined sequentially over a 7-day period. Virus was detected by means of immunohistochemistry and in situ hybridization in all tissues containing histologic lesions. Viral antigen and mRNA were observed initially in olfactory epithelium neurons, followed by olfactory bulbs and more caudal olfactory pathways in the brain. Virus also was detected throughout the ventricular system in the brain and central canal of the spinal cord. These results support both viral retrograde transneuronal transport and viral spread within the ventricular system. Other tissues containing viral antigen included airway epithelium and macrophages in the lungs, cardiac myocytes, and macrophages in cervical lymph nodes. In a second experiment, 15 adult, 20 juvenile, and 16 nestling deer mice were inoculated intradermally with VSV-NJ. Adults were refractory to infection by this route; however, nestlings and juveniles developed disseminated central nervous system infections. Viral antigen also was detected in cardiac myocytes and lymph node macrophages in these animals. Viremia was detected by virus isolation in 35/72 (49%) intranasally inoculated juvenile and adult mice and in 17/36 (47%) intradermally inoculated nestlings and juveniles from day 1 to day 3 postinoculation. The documentation of viremia in these animals suggests that they may have a role in the epidemiology of vector-borne vesicular stomatitis.

  9. Leucine-rich repeat-containing G protein-coupled receptor 4 facilitates vesicular stomatitis virus infection by binding vesicular stomatitis virus glycoprotein.

    Science.gov (United States)

    Zhang, Na; Huang, Hongjun; Tan, Binghe; Wei, Yinglei; Xiong, Qingqing; Yan, Yan; Hou, Lili; Wu, Nannan; Siwko, Stefan; Cimarelli, Andrea; Xu, Jianrong; Han, Honghui; Qian, Min; Liu, Mingyao; Du, Bing

    2017-10-06

    Vesicular stomatitis virus (VSV) and rabies and Chandipura viruses belong to the Rhabdovirus family. VSV is a common laboratory virus to study viral evolution and host immune responses to viral infection, and recombinant VSV-based vectors have been widely used for viral oncolysis, vaccination, and gene therapy. Although the tropism of VSV is broad, and its envelope glycoprotein G is often used for pseudotyping other viruses, the host cellular components involved in VSV infection remain unclear. Here, we demonstrate that the host protein leucine-rich repeat-containing G protein-coupled receptor 4 (Lgr4) is essential for VSV and VSV-G pseudotyped lentivirus (VSVG-LV) to infect susceptible cells. Accordingly, Lgr4-deficient mice had dramatically decreased VSV levels in the olfactory bulb. Furthermore, Lgr4 knockdown in RAW 264.7 cells also significantly suppressed VSV infection, and Lgr4 overexpression in RAW 264.7 cells enhanced VSV infection. Interestingly, only VSV infection relied on Lgr4, whereas infections with Newcastle disease virus, influenza A virus (A/WSN/33), and herpes simplex virus were unaffected by Lgr4 status. Of note, assays of virus entry, cell ELISA, immunoprecipitation, and surface plasmon resonance indicated that VSV bound susceptible cells via the Lgr4 extracellular domain. Pretreating cells with an Lgr4 antibody, soluble LGR4 extracellular domain, or R-spondin 1 blocked VSV infection by competitively inhibiting VSV binding to Lgr4. Taken together, the identification of Lgr4 as a VSV-specific host factor provides important insights into understanding VSV entry and its pathogenesis and lays the foundation for VSV-based gene therapy and viral oncolytic therapeutics. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  10. Antibodies against vesicular stomatitis virus in horses from southern, midwestern and northeastern Brazilian States

    Directory of Open Access Journals (Sweden)

    Vinícius Leobet Lunkes

    2016-08-01

    Full Text Available ABSTRACT: Vesicular stomatitis virus (VSV is the agent of a vesicular disease that affects many animal species and may be clinically confounded with foot-and-mouth disease in ruminant and swine. Horses are especially susceptible to VSV and may serve as sentinels for virus circulation. The present study investigated the presence of neutralizing antibodies against VSV Indiana III (VSIV-3 in serum samples of 3,626 horses from six states in three Brazilian regions: Southern (RS, n = 1,011, Midwest (GO/DF, n = 1,767 and Northeast (PB, PE, RN and CE, n = 848 collected between 2013 and 2014. Neutralizing antibodies against VSIV-3 (titers ≥40 were detected in 641 samples (positivity of 17.7%; CI95%:16.5-19.0%, being 317 samples from CE (87.3%; CI95%: 83.4-90.5 %; 109 from RN (65.7%; CI95%: 57.8 -72.7%; 124 from PB (45.4%; CI95%: 39.4-51.5%; 78 from GO/DF (4.4%; CI95%: 3.5-5.5% and nine samples of RS (0.9%; CI95%: 0.4-1.7%. Several samples from the Northeast and Midwest harbored high neutralizing titers, indicating a recent exposure to the virus. In contrast, samples from RS had low titers, possibly due to a past remote exposure. Several positive samples presented neutralizing activity against other VSV serotypes (Indiana I and New Jersey, yet in lower titers, indicating the specificity of the response to VSIV-3. These results demonstrated a relatively recent circulation of VSIV-3 in northeastern Brazilian States, confirming clinical findings and demonstrating the sanitary importance of this infection.

  11. Changes in Susceptibility to Oncolytic Vesicular Stomatitis Virus during Progression of Prostate Cancer.

    Science.gov (United States)

    Yu, Nanmeng; Puckett, Shelby; Antinozzi, Peter A; Cramer, Scott D; Lyles, Douglas S

    2015-05-01

    A major challenge to oncolytic virus therapy is that individual cancers vary in their sensitivity to oncolytic viruses, even when these cancers arise from the same tissue type. Variability in response may arise due to differences in the initial genetic lesions leading to cancer development. Alternatively, susceptibility to viral oncolysis may change during cancer progression. These hypotheses were tested using cells from a transgenic mouse model of prostate cancer infected with vesicular stomatitis virus (VSV). Primary cultures from murine cancers derived from prostate-specific Pten deletion contained a mixture of cells that were susceptible and resistant to VSV. Castration-resistant cancers contained a higher percentage of susceptible cells than cancers from noncastrated mice. These results indicate both susceptible and resistant cells can evolve within the same tumor. The role of Pten deletion was further investigated using clonal populations of murine prostate epithelial (MPE) progenitor cells and tumor-derived Pten(-/-) cells. Deletion of Pten in MPE progenitor cells using a lentivirus vector resulted in cells that responded poorly to interferon and were susceptible to VSV infection. In contrast, tumor-derived Pten(-/-) cells expressed higher levels of the antiviral transcription factor STAT1, activated STAT1 in response to VSV, and were resistant to VSV infection. These results suggest that early in tumor development following Pten deletion, cells are primarily sensitive to VSV, but subsequent evolution in tumors leads to development of cells that are resistant to VSV infection. Further evolution in castration-resistant tumors leads to tumors in which cells are primarily sensitive to VSV. There has been a great deal of progress in the development of replication-competent viruses that kill cancer cells (oncolytic viruses). However, a major problem is that individual cancers vary in their sensitivity to oncolytic viruses, even when these cancers arise from the

  12. Tumor Necrosis Factor-Mediated Survival of CD169+ Cells Promotes Immune Activation during Vesicular Stomatitis Virus Infection

    DEFF Research Database (Denmark)

    Shinde, Prashant V; Xu, Haifeng C; Maney, Sathish Kumar

    2018-01-01

    Innate immune activation is essential to mount an effective antiviral response and to prime adaptive immunity. Although a crucial role of CD169(+) cells during vesicular stomatitis virus (VSV) infections is increasingly recognized, factors regulating CD169(+) cells during viral infections remain...... defense against viral pathogens. CD169(+) macrophages are shown to activate innate and adaptive immunity via "enforced virus replication" a controlled amplification of virus particles. However, factors regulating the CD169(+) macrophages remain to be studied. In this paper, we show that after Vesicular...... stomatitis virus infection, phagocytes produce tumor necrosis factor (TNF) which signals via TNFR1 and promote "enforced virus replication" in CD169(+) macrophages. Consequently, lack of TNF or TNFR1 resulted in defective immune activation and VSV clearance....

  13. Some Attenuated Variants of Vesicular Stomatitis Virus Show Enhanced Oncolytic Activity against Human Glioblastoma Cells relative to Normal Brain Cells▿

    OpenAIRE

    Wollmann, Guido; Rogulin, Vitaliy; Simon, Ian; Rose, John K.; van den Pol, Anthony N.

    2009-01-01

    Vesicular stomatitis virus (VSV) has been shown in laboratory studies to be effective against a variety of tumors, including malignant brain tumors. However, attenuation of VSV may be necessary to balance the potential toxicity toward normal cells, particularly when targeting brain tumors. Here we compared 10 recombinant VSV variants resulting from different attenuation strategies. Attenuations included gene shifting (VSV-p1-GFP/RFP), M protein mutation (VSV-M51), G protein cytoplasmic tail t...

  14. Pseudotyping Vesicular Stomatitis Virus with Lymphocytic Choriomeningitis Virus Glycoproteins Enhances Infectivity for Glioma Cells and Minimizes Neurotropism▿†

    OpenAIRE

    Muik, Alexander; Kneiske, Inna; Werbizki, Marina; Wilflingseder, Doris; Giroglou, Tsanan; Ebert, Oliver; Kraft, Anna; Dietrich, Ursula; Zimmer, Gert; Momma, Stefan; von Laer, Dorothee

    2011-01-01

    Vesicular stomatitis virus (VSV)-based oncolytic virotherapy has the potential to significantly improve the prognosis of aggressive malignancies such as brain cancer. However, VSV's inherent neurotoxicity has hindered clinical development so far. Given that this neurotropism is attributed to the glycoprotein VSV-G, VSV was pseudotyped with the nonneurotropic envelope glycoprotein of the lymphocytic choriomeningitis virus (LCMV-GP→VSV-GP). Compared to VSV, VSV-GP showed enhanced infectivity fo...

  15. Evaluation of the Protective Efficacy of Recombinant Vesicular Stomatitis Virus Vectors Against Marburg Hemorrhagic Fever in Nonhuman Primate Models

    Science.gov (United States)

    2007-01-19

    VSV (Simon, Cardomone et al. 1990), borna disease virus (Formella, Jehle et al. 2000), and Sinbis virus (Karpf, Lenches et al. 1997). The...C., et al. (2000). "Sequence variability of Borna disease virus : resistance to superinfection may contribute to high genome stability in...Marburg virus disease ". S Afr Med J 66(2):50-4 Roberts, A., L. Buonocore, et al. (1999). "Attenuated vesicular stomatitis viruses as vaccine vectors." J

  16. [Antitumor effects of matrix protein of vesicular stomatic virus on EL4 lymphoma mice].

    Science.gov (United States)

    Lin, Shi-jia; Yu, Qin-mei; Meng, Wen-tong; Wen, Yan-jun; Chen, Li-juan; Niu, Ting

    2011-03-01

    To explore antitumor effects of plasmid pcDNA3. 1-MP encoding matrix protein of vesicular stomatitis virus (VSV) complexed with cationic liposome (DOTAP:CHOL) in mice with EL4 lymphoma. C57BL/6 mouse model with EL4 lymphoma was established. Sixty mice bearing EL4 lymphoma were divided randomly into five groups including Lip-MP, Lip-pVAX, Lip, ADM and NS groups, which were intravenously injected with liposome-pcDNA 3. 1-MP complex, liposome-pVAX complex, empty liposome, Adriamycin and normal saline respectively every three days. Tumor volumes and survival time were monitored. Microvessel density and tumor proliferative index in tumor tissues were determined by CD31, Ki-67 immunohistochemistry staining, meanwhile the tumor apoptosis index was measured by TUNEL method. From 6 days after treatments on, the tumor volume in Lip-MP group was much smaller than that in Lip-pVAX, Lip and NS group (P EL4 tumor cells in vivo (P EL4 lymphoma, which may be related to the induction of tumor cell apoptosis, inhibition of tumor angiogenesis, and suppression of tumor cell proliferation.

  17. Structure of the L Protein of Vesicular Stomatitis Virus from Electron Cryomicroscopy.

    Science.gov (United States)

    Liang, Bo; Li, Zongli; Jenni, Simon; Rahmeh, Amal A; Morin, Benjamin M; Grant, Timothy; Grigorieff, Nikolaus; Harrison, Stephen C; Whelan, Sean P J

    2015-07-16

    The large (L) proteins of non-segmented, negative-strand RNA viruses, a group that includes Ebola and rabies viruses, catalyze RNA-dependent RNA polymerization with viral ribonucleoprotein as template, a non-canonical sequence of capping and methylation reactions, and polyadenylation of viral messages. We have determined by electron cryomicroscopy the structure of the vesicular stomatitis virus (VSV) L protein. The density map, at a resolution of 3.8 Å, has led to an atomic model for nearly all of the 2109-residue polypeptide chain, which comprises three enzymatic domains (RNA-dependent RNA polymerase [RdRp], polyribonucleotidyl transferase [PRNTase], and methyltransferase) and two structural domains. The RdRp resembles the corresponding enzymatic regions of dsRNA virus polymerases and influenza virus polymerase. A loop from the PRNTase (capping) domain projects into the catalytic site of the RdRp, where it appears to have the role of a priming loop and to couple product elongation to large-scale conformational changes in L. Copyright © 2015 Elsevier Inc. All rights reserved.

  18. Structure of the L-protein of vesicular stomatitis virus from electron cryomicroscopy

    Science.gov (United States)

    Liang, Bo; Li, Zongli; Jenni, Simon; Rahmeh, Amal A.; Morin, Benjamin M.; Grant, Tim; Grigorieff, Nikolaus; Harrison, Stephen C.; Whelan, Sean P.J.

    2015-01-01

    The large (L) proteins of non-segmented, negative-strand RNA viruses, a group that includes Ebola and rabies viruses, catalyze RNA-dependent RNA polymerization with viral ribonucleoprotein as template, a noncanonical sequence of capping and methylation reactions, and polyadenylation of viral messages. We have determined by electron cryomicroscopy the structure of the vesicular stomatitis virus (VSV) L protein. The density map, at a resolution of 3.8 Å, has led to an atomic model for nearly all of the 2109-residue polypeptide chain, which comprises three enzymatic domains [RNA-dependent RNA polymerase (RdRp), polyribonucleotidyl transferase (PRNTase), and methyl transferase] and two structural domains. The RdRp resembles the corresponding enzymatic regions of dsRNA virus polymerases and influenza virus polymerase. A loop from the PRNTase (capping) domain projects into the catalytic site of the RdRp, where it appears to have the role of a priming loop and to couple product elongation to large-scale conformational changes in L. PMID:26144317

  19. Vesicular stomatitis virus as a flexible platform for oncolytic virotherapy against cancer

    Science.gov (United States)

    Hastie, Eric

    2012-01-01

    Oncolytic virus (OV) therapy is an emerging anti-cancer approach that utilizes viruses to preferentially infect and kill cancer cells, while not harming healthy cells. Vesicular stomatitis virus (VSV) is a prototypic non-segmented, negative-strand RNA virus with inherent OV qualities. Antiviral responses induced by type I interferon pathways are believed to be impaired in most cancer cells, making them more susceptible to VSV than normal cells. Several other factors make VSV a promising OV candidate for clinical use, including its well-studied biology, a small, easily manipulated genome, relative independence of a receptor or cell cycle, cytoplasmic replication without risk of host-cell transformation, and lack of pre-existing immunity in humans. Moreover, various VSV-based recombinant viruses have been engineered via reverse genetics to improve oncoselectivity, safety, oncotoxicity and stimulation of tumour-specific immunity. Alternative delivery methods are also being studied to minimize premature immune clearance of VSV. OV treatment as a monotherapy is being explored, although many studies have employed VSV in combination with radiotherapy, chemotherapy or other OVs. Preclinical studies with various cancers have demonstrated that VSV is a promising OV; as a result, a human clinical trial using VSV is currently in progress. PMID:23052398

  20. PEGylation of Vesicular Stomatitis Virus Extends Virus Persistence in Blood Circulation of Passively Immunized Mice

    Science.gov (United States)

    Tesfay, Mulu Z.; Kirk, Amber C.; Hadac, Elizabeth M.; Griesmann, Guy E.; Federspiel, Mark J.; Barber, Glen N.; Henry, Stephen M.; Peng, Kah-Whye

    2013-01-01

    We are developing oncolytic vesicular stomatitis viruses (VSVs) for systemic treatment of multiple myeloma, an incurable malignancy of antibody-secreting plasma cells that are specifically localized in the bone marrow. One of the presumed advantages for using VSV as an oncolytic virus is that human infections are rare and preexisting anti-VSV immunity is typically lacking in cancer patients, which is very important for clinical success. However, our studies show that nonimmune human and mouse serum can neutralize clinical-grade VSV, reducing the titer by up to 4 log units in 60 min. In addition, we show that neutralizing anti-VSV antibodies negate the antitumor efficacy of VSV, a concern for repeat VSV administration. We have investigated the potential use of covalent modification of VSV with polyethylene glycol (PEG) or a function-spacer-lipid (FSL)–PEG construct to inhibit serum neutralization and to limit hepatosplenic sequestration of systemically delivered VSV. We report that in mice passively immunized with neutralizing anti-VSV antibodies, PEGylation of VSV improved the persistence of VSV in the blood circulation, maintaining a more than 1-log-unit increase in VSV genome copies for up to 1 h compared to the genome copy numbers for the non-PEGylated virus, which was mostly cleared within 10 min after intravenous injection. We are currently investigating if this increase in PEGylated VSV circulating half-life can translate to increased virus delivery and better efficacy in mouse models of multiple myeloma. PMID:23325695

  1. Vesicular stomatitis virus infection promotes immune evasion by preventing NKG2D-ligand surface expression.

    Directory of Open Access Journals (Sweden)

    Helle Jensen

    Full Text Available Vesicular stomatitis virus (VSV has recently gained attention for its oncolytic ability in cancer treatment. Initially, we hypothesized that VSV infection could increase immune recognition of cancer cells through induction of the immune stimulatory NKG2D-ligands. Here we show that VSV infection leads to a robust induction of MICA mRNA expression, however the subsequent surface expression is potently hindered. Thus, VSV lines up with human cytomegalovirus (HCMV and adenovirus, which actively subvert the immune system by negatively affecting NKG2D-ligand surface expression. VSV infection caused an active suppression of NKG2D-ligand surface expression, affecting both endogenous and histone deacetylase (HDAC-inhibitor induced MICA, MICB and ULBP-2 expression. The classical immune escape mechanism of VSV (i.e., the M protein blockade of nucleocytoplasmic mRNA transport was not involved, as the VSV mutant strain, VSV(ΔM51, which possess a defective M protein, prevented MICA surface expression similarly to wild-type VSV. The VSV mediated down modulation of NKG2D-ligand expression did not involve apoptosis. Constitutive expression of MICA bypassed the escape mechanism, suggesting that VSV affect NKG2D-ligand expression at an early post-transcriptional level. Our results show that VSV possess an escape mechanism, which could affect the immune recognition of VSV infected cancer cells. This may also have implications for immune recognition of cancer cells after combined treatment with VSV and chemotherapeutic drugs.

  2. Vesicular stomatitis virus polymerase's strong affinity to its template suggests exotic transcription models.

    Directory of Open Access Journals (Sweden)

    Xiaolin Tang

    2014-12-01

    Full Text Available Vesicular stomatitis virus (VSV is the prototype for negative sense non segmented (NNS RNA viruses which include potent human and animal pathogens such as Rabies, Ebola and measles. The polymerases of NNS RNA viruses only initiate transcription at or near the 3' end of their genome template. We measured the dissociation constant of VSV polymerases from their whole genome template to be 20 pM. Given this low dissociation constant, initiation and sustainability of transcription becomes nontrivial. To explore possible mechanisms, we simulated the first hour of transcription using Monte Carlo methods and show that a one-time initial dissociation of all polymerases during entry is not sufficient to sustain transcription. We further show that efficient transcription requires a sliding mechanism for non-transcribing polymerases and can be realized with different polymerase-polymerase interactions and distinct template topologies. In conclusion, we highlight a model in which collisions between transcribing and sliding non-transcribing polymerases result in release of the non-transcribing polymerases allowing for redistribution of polymerases between separate templates during transcription and suggest specific experiments to further test these mechanisms.

  3. Inducible vesicular stomatitis virus (VSV) L cell line for packaging of recombinant VSV.

    Science.gov (United States)

    Hong, Seong-Karp; Jung, Yong-Tae; Park, Seung-Won; Paik, Soon-Young

    2005-10-01

    Recently, recombinant vesicular stomatitis viruses (VSV) have been developed as high-level expression vectors which serve as effective vaccine vectors in animals. An ideal approach for VSV vector production would be the development of stable packaging cell lines that can produce vector particles without transfection step. In this report, we describe generation of an inducible cell line that expresses the VSV polymerase gene (L) under the control of the reverse tetracycline-controlled transactivator (rtTA) system as a first step to make VSV-based packaging cell lines. Integrated polymerase (L) gene was controlled by an rtetR-dependent promoter in the rtTA-producing BHK cell line. When the cell lines were cultured in the presence of tet (tetracycline) or tetracycline derivative doxycycline, the recombinant VSV and wild type VSV were replicated, whereas in the absence of tet or tetracycline derivative doxycycline, the recombinant VSV was not replicated. Viral supernatants were harvested, diluted, and monitored by plaque assay for the presence of infectious VSV. Plaques of VSV containing an additional sequence encoding the EGFP protein allowed rapid detection of infection. Our results suggest wide applications of other surrogate viruses based on VSV. The availability of stable packaging cell lines represents a step toward the use of a VSV vector delivery system that can allow scale-up production of vector-stocks for gene therapy.

  4. [Vesicular stomatitis virus (VSV) as a vaccine vector for immunization against viral infections].

    Science.gov (United States)

    Tomczyk, Tomasz; Orzechowska, Beata

    2013-01-11

    Vesicular stomatitis virus (VSV), a member of the Rhabdoviridae family, is a promising candidate for potential use in construction of antiviral vaccines. In the natural environment VSV is a pathogen of wild ungulates and livestock. Some of the features that make VSV an excellent platform for the development of a range of viral therapeutics includes its immunogenicity and ability to grow to high titers in cell lines approved for vaccine use. Infection in humans is rare and usually asymptomatic, with mild flu-like symptoms. Moreover, due to affinity of VSV envelope glycoprotein to the LDL (low-density lipoprotein) receptor, VSV is effective at targeting a variety of tissues in vivo. A series of research results confirm the possibility of developing VSV-based vaccines against human papilloma viruses (HPV), human immunodeficiency virus (HIV), hepatitis B virus (HBV) and filoviruses (MARV, ZEBOV and SEBOV), as well as the potential use of a successfully developed vaccine against hepatitis C virus (HCV). VSV is neurotropic and infection can cause a viral encephalitis in experimental animals. Therefore, intensive studies are being undertaken to achieve satisfactory expression of the viral antigens while maintaining the safety of the constructed vectors.

  5. Oncolytic vesicular stomatitis virus expressing interferon-σ has enhanced therapeutic activity

    Directory of Open Access Journals (Sweden)

    Marie-Claude Bourgeois-Daigneault

    2016-01-01

    Full Text Available Oncolytic viruses are known to stimulate the antitumor immune response by specifically replicating in tumor cells. This is believed to be an important aspect of the durable responses observed in some patients and the field is rapidly moving toward immunotherapy. As a further means to engage the immune system, we engineered a virus, vesicular stomatitis virus (VSV, to encode the proinflammatory cytokine interferon-σ. We used the 4T1 mammary adenocarcinoma as well as other murine tumor models to characterize immune responses in tumor-bearing animals generated by treatment with our viruses. The interferon-σ-encoding virus demonstrated greater activation of dendritic cells and drove a more profound secretion of proinflammatory cytokines compared to the parental virus. From a therapeutic point of view, the interferon-σ virus slowed tumor growth, minimized lung tumors, and prolonged survival in several murine tumor models. The improved efficacy was lost in immunocompromized animals; hence the mechanism appears to be T-cell-mediated. Taken together, these results demonstrate the ability of oncolytic viruses to act as immune stimulators to drive antitumor immunity as well as their potential for targeted gene therapy.

  6. Different effect of proteasome inhibition on vesicular stomatitis virus and poliovirus replication.

    Directory of Open Access Journals (Sweden)

    Nickolay Neznanov

    2008-04-01

    Full Text Available Proteasome activity is an important part of viral replication. In this study, we examined the effect of proteasome inhibitors on the replication of vesicular stomatitis virus (VSV and poliovirus. We found that the proteasome inhibitors significantly suppressed VSV protein synthesis, virus accumulation, and protected infected cells from toxic effect of VSV replication. In contrast, poliovirus replication was delayed, but not diminished in the presence of the proteasome inhibitors MG132 and Bortezomib. We also found that inhibition of proteasomes stimulated stress-related processes, such as accumulation of chaperone hsp70, phosphorylation of eIF2alpha, and overall inhibition of translation. VSV replication was sensitive to this stress with significant decline in replication process. Poliovirus growth was less sensitive with only delay in replication. Inhibition of proteasome activity suppressed cellular and VSV protein synthesis, but did not reduce poliovirus protein synthesis. Protein kinase GCN2 supported the ability of proteasome inhibitors to attenuate general translation and to suppress VSV replication. We propose that different mechanisms of translational initiation by VSV and poliovirus determine their sensitivity to stress induced by the inhibition of proteasomes. To our knowledge, this is the first study that connects the effect of stress induced by proteasome inhibition with the efficiency of viral infection.

  7. Vesicular Stomatitis Virus Variants Selectively Infect and Kill Human Melanomas but Not Normal Melanocytes

    Science.gov (United States)

    Wollmann, Guido; Davis, John N.; Bosenberg, Marcus W.

    2013-01-01

    Metastatic malignant melanoma remains one of the most therapeutically challenging forms of cancer. Here we test replication-competent vesicular stomatitis viruses (VSV) on 19 primary human melanoma samples and compare these infections with those of normal human melanocyte control cells. Even at a low viral concentration, we found a strong susceptibility to viral oncolysis in over 70% of melanomas. In contrast, melanocytes displayed strong resistance to virus infection and showed complete protection by interferon. Several recombinant VSVs were compared, and all infected and killed most melanomas with differences in the time course with increasing rates of melanoma infection, as follows: VSV-CT9-M51 VSV-M51 VSV-G/GFP VSV-rp30. VSV-rp30 sequencing revealed 2 nonsynonymous mutations at codon positions P126 and L223, both of which appear to be required for the enhanced phenotype. VSV-rp30 showed effective targeting and infection of multiple subcutaneous and intracranial melanoma xenografts in SCID mice after tail vein virus application. Sequence analysis of mutations in the melanomas used revealed that BRAF but not NRAS gene mutation status was predictive for enhanced susceptibility to infection. In mouse melanoma models with specific induced gene mutations including mutations of the Braf, Pten, and Cdkn2a genes, viral infection correlated with the extent of malignant transformation. Similar to human melanocytes, mouse melanocytes resisted VSV-rp30 infection. This study confirms the general susceptibility of the majority of human melanoma types for VSV-mediated oncolysis. PMID:23552414

  8. Vesicular stomatitis virus as an oncolytic agent against pancreatic ductal adenocarcinoma.

    Science.gov (United States)

    Murphy, Andrea M; Besmer, Dahlia M; Moerdyk-Schauwecker, Megan; Moestl, Natascha; Ornelles, David A; Mukherjee, Pinku; Grdzelishvili, Valery Z

    2012-03-01

    Vesicular stomatitis virus (VSV) is a promising oncolytic agent against a variety of cancers. However, it has never been tested in any pancreatic cancer model. Pancreatic ductal adenocarcinoma (PDA) is the most common and aggressive form of pancreatic cancer. In this study, the oncolytic potentials of several VSV variants were analyzed in a panel of 13 clinically relevant human PDA cell lines and compared to conditionally replicative adenoviruses (CRAds), Sendai virus and respiratory syncytial virus. VSV variants showed oncolytic abilities superior to those of other viruses, and some cell lines that exhibited resistance to other viruses were successfully killed by VSV. However, PDA cells were highly heterogeneous in their susceptibility to virus-induced oncolysis, and several cell lines were resistant to all tested viruses. Resistant cells showed low levels of very early VSV RNA synthesis, indicating possible defects at initial stages of infection. In addition, unlike permissive PDA cell lines, most of the resistant cell lines were able to both produce and respond to interferon, suggesting that intact type I interferon responses contributed to their resistance phenotype. Four cell lines that varied in their permissiveness to VSV-ΔM51 and CRAd dl1520 were tested in mice, and the in vivo results closely mimicked those in vitro. While our results demonstrate that VSV is a promising oncolytic agent against PDA, further studies are needed to better understand the molecular mechanisms of resistance of some PDAs to oncolytic virotherapy.

  9. Repeatable population dynamics among vesicular stomatitis virus lineages evolved under high co-infection

    Directory of Open Access Journals (Sweden)

    Elizabeth S.C.P. Williams

    2016-03-01

    Full Text Available Parasites and hosts can experience oscillatory cycles, where the densities of these interacting species dynamically fluctuate through time. Viruses with different replication strategies can also interact to produce cyclical dynamics. Frequent cellular co-infection can select for defective-interfering particles (DIPs: cheater viruses with shortened genomes that interfere with intracellular replication of full-length (ordinary viruses. DIPs are positively selected when rare because they out-replicate ordinary viruses during co-infection, but DIPs are negatively selected when common because ordinary viruses become unavailable for intracellular exploitation via cheating. Here we tested whether oscillatory dynamics of ordinary viruses were similar across independently evolved populations of vesicular stomatitis virus (VSV. Results showed identical cyclical dynamics across populations in the first 10 experimental passages, which transitioned to repeatable dampened oscillations by passage 20. Genomic analyses revealed parallel molecular substitutions across populations, particularly novel mutations that became dominant by passage 10. Our study showed that oscillatory dynamics and molecular evolution of interacting viruses were highly repeatable in VSV populations passaged under frequent co-infection. Furthermore, our data suggested that frequent co-infection with DIPs caused lowered performance of full-length viruses, by reducing their population densities by orders of magnitude compared to reproduction of ordinary viruses during strictly clonal infections.

  10. Vesicular stomatitis virus variants selectively infect and kill human melanomas but not normal melanocytes.

    Science.gov (United States)

    Wollmann, Guido; Davis, John N; Bosenberg, Marcus W; van den Pol, Anthony N

    2013-06-01

    Metastatic malignant melanoma remains one of the most therapeutically challenging forms of cancer. Here we test replication-competent vesicular stomatitis viruses (VSV) on 19 primary human melanoma samples and compare these infections with those of normal human melanocyte control cells. Even at a low viral concentration, we found a strong susceptibility to viral oncolysis in over 70% of melanomas. In contrast, melanocytes displayed strong resistance to virus infection and showed complete protection by interferon. Several recombinant VSVs were compared, and all infected and killed most melanomas with differences in the time course with increasing rates of melanoma infection, as follows: VSV-CT9-M51 VSV-M51 VSV-G/GFP VSV-rp30. VSV-rp30 sequencing revealed 2 nonsynonymous mutations at codon positions P126 and L223, both of which appear to be required for the enhanced phenotype. VSV-rp30 showed effective targeting and infection of multiple subcutaneous and intracranial melanoma xenografts in SCID mice after tail vein virus application. Sequence analysis of mutations in the melanomas used revealed that BRAF but not NRAS gene mutation status was predictive for enhanced susceptibility to infection. In mouse melanoma models with specific induced gene mutations including mutations of the Braf, Pten, and Cdkn2a genes, viral infection correlated with the extent of malignant transformation. Similar to human melanocytes, mouse melanocytes resisted VSV-rp30 infection. This study confirms the general susceptibility of the majority of human melanoma types for VSV-mediated oncolysis.

  11. Preimplantation bovine embryos: Pathobiology of Haemophilus somnus exposure and resistance mechanisms to vesicular stomatitis virus

    Energy Technology Data Exchange (ETDEWEB)

    Thomson, M.S.

    1988-01-01

    Preimplantation bovine embryos were exposed in vitro to H. somnus to determine if the bacteria would adhere to zona pellucida-intact (ZP-I) embryos or adhere to or infect ZP-free embryos. The effect of H. somnus on embryonic development in vitro was also investigated. Electrophoretic comparisons of outer membrane proteins of H. somnus revealed 2 major protein bands common to 10 H. somnus isolates. A monoclonal antibody produced against the outer membrane proteins reacted to one of the major protein bands. The sensitivity of a nucleic acid probe for detection of vesicular stomatitis virus (VSV) was validated in cells in culture and used to determine if the synthetic double-stranded complex of polyriboinosinic and polyribocytidylic acids (poly I:C) would induce viral resistance in cultured bovine embryos. Two {sup 32}P-nick translated probes of high specific activity prepared from plasmids containing nucleic acid sequences of VSV virus were employed for viral mRNA detection in the tissue culture cells using a DNA-hybridization dot-blot technique. Using one of the probes, the technique was applied to detect differences in viral replication between four groups of bovine embryos (nonexposed, exposed to VSV virus, poly I:C-treated, and poly I:C-treated and exposed to VSV). The nucleic acid probe was sufficiently sensitive to detect differences in quantities of VSV mRNA among embryo treatment groups, resulting in the demonstration that resistance to viral infection was induced in day 9 bovine embryos.

  12. Recent advances in vesicular stomatitis virus-based oncolytic virotherapy: a 5-year update.

    Science.gov (United States)

    Felt, Sébastien A; Grdzelishvili, Valery Z

    2017-11-16

    Oncolytic virus (OV) therapy is an anti-cancer approach that uses viruses that preferentially infect, replicate in and kill cancer cells. Vesicular stomatitis virus (VSV, a rhabdovirus) is an OV that is currently being tested in the USA in several phase I clinical trials against different malignancies. Several factors make VSV a promising OV: lack of pre-existing human immunity against VSV, a small and easy to manipulate genome, cytoplasmic replication without risk of host cell transformation, independence of cell cycle and rapid growth to high titres in a broad range of cell lines facilitating large-scale virus production. While significant advances have been made in VSV-based OV therapy, room for improvement remains. Here we review recent studies (published in the last 5 years) that address 'old' and 'new' challenges of VSV-based OV therapy. These studies focused on improving VSV safety, oncoselectivity and oncotoxicity; breaking resistance of some cancers to VSV; preventing premature clearance of VSV; and stimulating tumour-specific immunity. Many of these approaches were based on combining VSV with other therapeutics. This review also discusses another rhabdovirus closely related to VSV, Maraba virus, which is currently being tested in Canada in phase I/II clinical trials.

  13. Genetic Inactivation of COPI Coatomer Separately Inhibits Vesicular Stomatitis Virus Entry and Gene Expression

    Science.gov (United States)

    Burdeinick-Kerr, Rebeca

    2012-01-01

    Viruses coopt cellular membrane transport to invade cells, establish intracellular sites of replication, and release progeny virions. Recent genome-wide RNA interference (RNAi) screens revealed that genetically divergent viruses require biosynthetic membrane transport by the COPI coatomer complex for efficient replication. Here we found that disrupting COPI function by RNAi inhibited an early stage of vesicular stomatitis virus (VSV) replication. To dissect which replication stage(s) was affected by coatomer inactivation, we used visual and biochemical assays to independently measure the efficiency of viral entry and gene expression in hamster (ldlF) cells depleted of the temperature-sensitive ε-COP subunit. We show that ε-COP depletion for 12 h caused a primary block to virus internalization and a secondary defect in viral gene expression. Using brefeldin A (BFA), a chemical inhibitor of COPI function, we demonstrate that short-term (1-h) BFA treatments inhibit VSV gene expression, while only long-term (12-h) treatments block virus entry. We conclude that prolonged coatomer inactivation perturbs cellular endocytic transport and thereby indirectly impairs VSV entry. Our results offer an explanation of why COPI coatomer is frequently identified in screens for cellular factors that support cell invasion by microbial pathogens. PMID:22072764

  14. Vesicular stomatitis virus enables gene transfer and transsynaptic tracing in a wide range of organisms

    Science.gov (United States)

    Mundell, Nathan A.; Beier, Kevin T.; Pan, Y. Albert; Lapan, Sylvain W.; Göz Aytürk, Didem; Berezovskii, Vladimir K.; Wark, Abigail R.; Drokhlyansky, Eugene; Bielecki, Jan; Born, Richard T.; Schier, Alexander F.

    2015-01-01

    Current limitations in technology have prevented an extensive analysis of the connections among neurons, particularly within nonmammalian organisms. We developed a transsynaptic viral tracer originally for use in mice, and then tested its utility in a broader range of organisms. By engineering the vesicular stomatitis virus (VSV) to encode a fluorophore and either the rabies virus glycoprotein (RABV‐G) or its own glycoprotein (VSV‐G), we created viruses that can transsynaptically label neuronal circuits in either the retrograde or anterograde direction, respectively. The vectors were investigated for their utility as polysynaptic tracers of chicken and zebrafish visual pathways. They showed patterns of connectivity consistent with previously characterized visual system connections, and revealed several potentially novel connections. Further, these vectors were shown to infect neurons in several other vertebrates, including Old and New World monkeys, seahorses, axolotls, and Xenopus. They were also shown to infect two invertebrates, Drosophila melanogaster, and the box jellyfish, Tripedalia cystophora, a species previously intractable for gene transfer, although no clear evidence of transsynaptic spread was observed in these species. These vectors provide a starting point for transsynaptic tracing in most vertebrates, and are also excellent candidates for gene transfer in organisms that have been refractory to other methods. J. Comp. Neurol. 523:1639–1663, 2015. © 2015 Wiley Periodicals, Inc. PMID:25688551

  15. Induction of apoptosis in pancreatic cancer cells by vesicular stomatitis virus.

    Science.gov (United States)

    Felt, Sébastien A; Moerdyk-Schauwecker, Megan J; Grdzelishvili, Valery Z

    2015-01-01

    Effective oncolytic virus (OV) therapy is dependent on the ability of replication-competent viruses to kill infected cancer cells. We previously showed that human pancreatic ductal adenocarcinoma (PDAC) cell lines are highly heterogeneous in their permissiveness to vesicular stomatitis virus (VSV), in part due to differences in type I interferon (IFN) signaling. Here, using 10 human PDAC cell lines and three different VSV recombinants (expressing ΔM51 or wild type matrix protein), we examined cellular and viral factors affecting VSV-mediated apoptosis activation in PDACs. In most cell lines, VSVs activated both extrinsic and intrinsic apoptosis pathways, and VSV-ΔM51 primarily activated the type II extrinsic pathway. In cells with defective IFN signaling, all VSV recombinants induced robust apoptosis, whereas VSV-ΔM51 was a more effective apoptosis activator in PDACs with virus-inducible IFN signaling. Three cell lines constitutively expressing high levels of IFN-stimulated genes (ISGs) were resistant to apoptosis under most experimental conditions, even when VSV replication levels were dramatically increased by Jak inhibitor I treatment. Two of these cell lines also poorly activated apoptosis when treated with Fas activating antibody, suggesting a general defect in apoptosis. Copyright © 2014 Elsevier Inc. All rights reserved.

  16. Durability of a vesicular stomatitis virus-based marburg virus vaccine in nonhuman primates.

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    Chad E Mire

    Full Text Available The filoviruses, Marburg virus (MARV and Ebola virus, causes severe hemorrhagic fever with high mortality in humans and nonhuman primates. A promising filovirus vaccine under development is based on a recombinant vesicular stomatitis virus (rVSV that expresses individual filovirus glycoproteins (GPs in place of the VSV glycoprotein (G. These vaccines have shown 100% efficacy against filovirus infection in nonhuman primates when challenge occurs 28-35 days after a single injection immunization. Here, we examined the ability of a rVSV MARV-GP vaccine to provide protection when challenge occurs more than a year after vaccination. Cynomolgus macaques were immunized with rVSV-MARV-GP and challenged with MARV approximately 14 months after vaccination. Immunization resulted in the vaccine cohort of six animals having anti-MARV GP IgG throughout the pre-challenge period. Following MARV challenge none of the vaccinated animals showed any signs of clinical disease or viremia and all were completely protected from MARV infection. Two unvaccinated control animals exhibited signs consistent with MARV infection and both succumbed. Importantly, these data are the first to show 100% protective efficacy against any high dose filovirus challenge beyond 8 weeks after final vaccination. These findings demonstrate the durability of VSV-based filovirus vaccines.

  17. Triptolide-mediated inhibition of interferon signaling enhances vesicular stomatitis virus-based oncolysis.

    Science.gov (United States)

    Ben Yebdri, Fethia; Van Grevenynghe, Julien; Tang, Vera A; Goulet, Marie-Line; Wu, Jian Hui; Stojdl, David F; Hiscott, John; Lin, Rongtuan

    2013-11-01

    Preclinical and clinical trials demonstrated that use of oncolytic viruses (OVs) is a promising new therapeutic approach to treat multiple types of cancer. To further improve their viral oncolysis, experimental strategies are now combining OVs with different cytotoxic compounds. In this study, we investigated the capacity of triptolide - a natural anticancer molecule - to enhance vesicular stomatitis virus (VSV) oncolysis in OV-resistant cancer cells. Triptolide treatment increased VSV replication in the human prostate cancer cell line PC3 and in other VSV-resistant cells in a dose- and time-dependent manner in vitro and in vivo. Mechanistically, triptolide (TPL) inhibited the innate antiviral response by blocking type I interferon (IFN) signaling, downstream of IRF3 activation. Furthermore, triptolide-enhanced VSV-induced apoptosis in a dose-dependent fashion in VSV-resistant cells, as measured by annexin-V, cleaved caspase-3, and B-cell lymphoma 2 staining. In vivo, using the TSA mammary adenocarcinoma and PC3 mouse xenograft models, combination treatment with VSV and triptolide delayed tumor growth and prolonged survival of tumor-bearing animals by enhancing viral replication. Together, these results demonstrate that triptolide inhibition of IFN production sensitizes prostate cancer cells to VSV replication and virus-mediated apoptosis.

  18. N-Myc expression enhances the oncolytic effects of vesicular stomatitis virus in human neuroblastoma cells

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    Juan C Corredor

    2016-01-01

    Full Text Available N-myc oncogene amplification is associated but not present in all cases of high-risk neuroblastoma (NB. Since oncogene expression could often modulate sensitivity to oncolytic viruses, we wanted to examine if N-myc expression status would determine virotherapy efficacy to high-risk NB. We showed that induction of exogenous N-myc in a non-N-myc-amplified cell line background (TET-21N increased susceptibility to oncolytic vesicular stomatitis virus (mutant VSVδM51 and alleviated the type I IFN-induced antiviral state. Cells with basal N-myc, on the other hand, were less susceptible to virus-induced oncolysis and established a robust IFN-mediated antiviral state. The same effects were also observed in NB cell lines with and without N-myc amplification. Microarray analysis showed that N-myc overexpression in TET-21N cells downregulated IFN-stimulated genes (ISGs with known antiviral functions. Furthermore, virus infection caused significant changes in global gene expression in TET-21N cells overexpressing N-myc. Such changes involved ISGs with various functions. Therefore, the present study showed that augmented susceptibility to VSVδM51 by N-myc at least involves downregulation of ISGs with antiviral functions and alleviation of the IFN-stimulated antiviral state. Our studies suggest the potential utility of N-myc amplification/overexpression as a predictive biomarker of virotherapy response for high-risk NB using IFN-sensitive oncolytic viruses.

  19. Recombinant vesicular stomatitis virus vaccine vectors expressing filovirus glycoproteins lack neurovirulence in nonhuman primates.

    Directory of Open Access Journals (Sweden)

    Chad E Mire

    Full Text Available The filoviruses, Marburg virus and Ebola virus, cause severe hemorrhagic fever with high mortality in humans and nonhuman primates. Among the most promising filovirus vaccines under development is a system based on recombinant vesicular stomatitis virus (rVSV that expresses an individual filovirus glycoprotein (GP in place of the VSV glycoprotein (G. The main concern with all replication-competent vaccines, including the rVSV filovirus GP vectors, is their safety. To address this concern, we performed a neurovirulence study using 21 cynomolgus macaques where the vaccines were administered intrathalamically. Seven animals received a rVSV vector expressing the Zaire ebolavirus (ZEBOV GP; seven animals received a rVSV vector expressing the Lake Victoria marburgvirus (MARV GP; three animals received rVSV-wild type (wt vector, and four animals received vehicle control. Two of three animals given rVSV-wt showed severe neurological symptoms whereas animals receiving vehicle control, rVSV-ZEBOV-GP, or rVSV-MARV-GP did not develop these symptoms. Histological analysis revealed major lesions in neural tissues of all three rVSV-wt animals; however, no significant lesions were observed in any animals from the filovirus vaccine or vehicle control groups. These data strongly suggest that rVSV filovirus GP vaccine vectors lack the neurovirulence properties associated with the rVSV-wt parent vector and support their further development as a vaccine platform for human use.

  20. Fabrication of Photofunctional Nanoporous Membrane and Its Photoinactivation Effect of Vesicular Stomatitis Virus

    Directory of Open Access Journals (Sweden)

    Kang-Kyun Wang

    2012-01-01

    Full Text Available Fabrication and photophysical study of photofunctional nanoporous alumina membrane (PNAM were performed, and its application of photodynamic antimicrobial chemotherapy (PACT was investigated. Nanoporous alumina membrane (NAM was fabricated by two-step aluminium anodic oxidation process. Surface of the fabricated NAM was modified with organo-silane agent to induce covalent bonding between NAM and a photosensitizer (PtCP: [5,10,15-triphenyl-20-(4-methoxycarbonylphenyl-porphyrin] platinum. PtCP was covalently bonded to the surface of the modified NAM by nucleophilic acyl substitution reaction process. The morphology and the photophysical properties of the fabricated PNAM were confirmed with field emission scanning electron microscope (FE-SEM, steady-state spectroscopies, and nanosecond laser-induced time-resolved spectroscopy. For the efficacy study of PNAM in PACT, an enveloped animal virus, vesicular stomatitis virus (VSV, was utilized as a target organism. Antiviral effect of the PNAM-PACT was measured by the extent of suppression of plaque-forming units (PFU after the light irradiation. In the cultures inoculated with PACT-treated VSV, the suppression of PFU was prominent, which demonstrates that PNAM is a potential bio clean-up tool.

  1. Vesicular stomatitis virus as a flexible platform for oncolytic virotherapy against cancer.

    Science.gov (United States)

    Hastie, Eric; Grdzelishvili, Valery Z

    2012-12-01

    Oncolytic virus (OV) therapy is an emerging anti-cancer approach that utilizes viruses to preferentially infect and kill cancer cells, while not harming healthy cells. Vesicular stomatitis virus (VSV) is a prototypic non-segmented, negative-strand RNA virus with inherent OV qualities. Antiviral responses induced by type I interferon pathways are believed to be impaired in most cancer cells, making them more susceptible to VSV than normal cells. Several other factors make VSV a promising OV candidate for clinical use, including its well-studied biology, a small, easily manipulated genome, relative independence of a receptor or cell cycle, cytoplasmic replication without risk of host-cell transformation, and lack of pre-existing immunity in humans. Moreover, various VSV-based recombinant viruses have been engineered via reverse genetics to improve oncoselectivity, safety, oncotoxicity and stimulation of tumour-specific immunity. Alternative delivery methods are also being studied to minimize premature immune clearance of VSV. OV treatment as a monotherapy is being explored, although many studies have employed VSV in combination with radiotherapy, chemotherapy or other OVs. Preclinical studies with various cancers have demonstrated that VSV is a promising OV; as a result, a human clinical trial using VSV is currently in progress.

  2. Chikungunya, Influenza, Nipah, and Semliki Forest Chimeric Viruses with Vesicular Stomatitis Virus: Actions in the Brain.

    Science.gov (United States)

    van den Pol, Anthony N; Mao, Guochao; Chattopadhyay, Anasuya; Rose, John K; Davis, John N

    2017-03-15

    Recombinant vesicular stomatitis virus (VSV)-based chimeric viruses that include genes from other viruses show promise as vaccines and oncolytic viruses. However, the critical safety concern is the neurotropic nature conveyed by the VSV glycoprotein. VSVs that include the VSV glycoprotein (G) gene, even in most recombinant attenuated strains, can still show substantial adverse or lethal actions in the brain. Here, we test 4 chimeric viruses in the brain, including those in which glycoprotein genes from Nipah, chikungunya (CHIKV), and influenza H5N1 viruses were substituted for the VSV glycoprotein gene. We also test a virus-like vesicle (VLV) in which the VSV glycoprotein gene is expressed from a replicon encoding the nonstructural proteins of Semliki Forest virus. VSVΔG-CHIKV, VSVΔG-H5N1, and VLV were all safe in the adult mouse brain, as were VSVΔG viruses expressing either the Nipah F or G glycoprotein. In contrast, a complementing pair of VSVΔG viruses expressing Nipah G and F glycoproteins were lethal within the brain within a surprisingly short time frame of 2 days. Intranasal inoculation in postnatal day 14 mice with VSVΔG-CHIKV or VLV evoked no adverse response, whereas VSVΔG-H5N1 by this route was lethal in most mice. A key immune mechanism underlying the safety of VSVΔG-CHIKV, VSVΔG-H5N1, and VLV in the adult brain was the type I interferon response; all three viruses were lethal in the brains of adult mice lacking the interferon receptor, suggesting that the viruses can infect and replicate and spread in brain cells if not blocked by interferon-stimulated genes within the brain.IMPORTANCE Vesicular stomatitis virus (VSV) shows considerable promise both as a vaccine vector and as an oncolytic virus. The greatest limitation of VSV is that it is highly neurotropic and can be lethal within the brain. The neurotropism can be mostly attributed to the VSV G glycoprotein. Here, we test 4 chimeric viruses of VSV with glycoprotein genes from Nipah

  3. mRNA cap methylation influences pathogenesis of vesicular stomatitis virus in vivo.

    Science.gov (United States)

    Ma, Yuanmei; Wei, Yongwei; Zhang, Xiaodong; Zhang, Yu; Cai, Hui; Zhu, Yang; Shilo, Konstantin; Oglesbee, Michael; Krakowka, Steven; Whelan, Sean P J; Li, Jianrong

    2014-03-01

    One role of mRNA cap guanine-N-7 (G-N-7) methylation is to facilitate the efficient translation of mRNA. The role of mRNA cap ribose 2'-O methylation is enigmatic, although recent work has implicated this as a signature to avoid detection of RNA by the innate immune system (S. Daffis, K. J. Szretter, J. Schriewer, J. Q. Li, S. Youn, J. Errett, T. Y. Lin, S. Schneller, R. Zust, H. P. Dong, V. Thiel, G. C. Sen, V. Fensterl, W. B. Klimstra, T. C. Pierson, R. M. Buller, M. Gale, P. Y. Shi, M. S. Diamond, Nature 468:452-456, 2010, doi:10.1038/nature09489). Working with vesicular stomatitis virus (VSV), we previously showed that a panel of recombinant VSVs carrying mutations at a predicted methyltransferase catalytic site (rVSV-K1651A, -D1762A, and -E1833Q) or S-adenosylmethionine (SAM) binding site (rVSV-G1670A, -G1672A, and -G4A) were defective in cap methylation and were also attenuated for growth in cell culture. Here, we analyzed the virulence of these recombinants in mice. We found that rVSV-K1651A, -D1762A, and -E1833Q, which are defective in both G-N-7 and 2'-O methylation, were highly attenuated in mice. All three viruses elicited a high level of neutralizing antibody and provided full protection against challenge with the virulent VSV. In contrast, mice inoculated with rVSV-G1670A and -G1672A, which are defective only in G-N-7 methylation, were attenuated in vivo yet retained a low level of virulence. rVSV-G4A, which is completely defective in both G-N-7 and 2'-O methylation, also exhibited low virulence in mice despite the fact that productive viral replication was not detected in lung and brain. Taken together, our results suggest that abrogation of viral mRNA cap methylation can serve as an approach to attenuate VSV, and perhaps other nonsegmented negative-strand RNA viruses, for potential application as vaccines and viral vectors. Nonsegmented negative-sense (NNS) RNA viruses include a wide range of significant human, animal, and plant pathogens. For many

  4. Highly Attenuated Recombinant Vesicular Stomatitis Virus VSV-12′GFP Displays Immunogenic and Oncolytic Activity

    Science.gov (United States)

    Davis, John N.

    2013-01-01

    Vesicular stomatitis virus (VSV) has shown considerable promise both as an immunization vector and as an oncolytic virus. In both applications, an important concern is the safety profile of the virus. To generate a highly attenuated virus, we added two reporter genes to the 3′ end of the VSV genome, thereby shifting the NPMGL genes from positions 1 to 5 to positions 3 to 7. The resulting virus (VSV-12′GFP) was highly attenuated, generating smaller plaques than four other attenuated VSVs. In one-step growth curves, VSV-12′GFP displayed the slowest growth kinetics. The mechanism of attenuation appears to be due to reduced expression of VSV genes downstream of the reporter genes, as suggested by a 10.4-fold reduction in L-protein RNA transcript. Although attenuated, VSV-12′GFP was highly effective at generating an immune response, indicated by a high-titer antibody response against the green fluorescent protein (GFP) expressed by the virus. Although VSV-12′GFP was more attenuated than other VSVs on both normal and cancer cells, it nonetheless showed a greater level of infection of human cancer cells (glioma and melanoma) than of normal cells, and this effect was magnified in glioma by interferon application, indicating selective oncolysis. Intravenous VSV-12′GFP selectively infected human gliomas implanted into SCID mice subcutaneously or intracranially. All postnatal day 16 mice given intranasal VSV-12′GFP survived, whereas only 10% of those given VSV-G/GFP survived, indicating reduced neurotoxicity. Intratumoral injection of tumors with VSV-12′GFP dramatically suppressed tumor growth and enhanced survival. Together these data suggest this recombinant virus merits further study for its oncolytic and vaccine potential. PMID:23135719

  5. In Vitro and In Vivo Attenuation of Vesicular Stomatitis Virus (VSV) by Phosphoprotein Deletion.

    Science.gov (United States)

    Wongthida, Phonphimon; Jengarn, Juggragarn; Narkpuk, Jaraspim; Koonyosying, Pongpisid; Srisutthisamphan, Kanjana; Wanitchang, Asawin; Leaungwutiwong, Pornsawan; Teeravechyan, Samaporn; Jongkaewwattana, Anan

    2016-01-01

    Vesicular stomatitis virus (VSV) is highly immunogenic and able to stimulate both innate and adaptive immune responses. However, its ability to induce adverse effects has held back the use of VSV as a potential vaccine vector. In this study we developed VSV-ΔP, a safe yet potent replication-defective recombinant VSV in which the phosphoprotein (P) gene was deleted. VSV-ΔP replicated only in supporting cells expressing P (BHK-P cells) and at levels more than 2 logs lower than VSV. In vivo studies indicated that the moderate replication of VSV-ΔP in vitro was associated with the attenuation of this virus in the mouse model, whereas mice intracranially injected with VSV succumbed to neurotoxicity. Furthermore, we constructed VSV and VSV-ΔP expressing a variety of antigens including hemagglutinin-neuraminidase (HN) from Newcastle disease virus (NDV), hemagglutinin (HA) from either a 2009 H1N1 pandemic influenza virus (pdm/09) or the avian H7N9. VSV and VSV-ΔP incorporated the foreign antigens on their surface resulting in induction of robust neutralizing antibody, serum IgG, and hemagglutination inhibition (HAI) titers against their corresponding viruses. These results indicated that VSV with P gene deletion was attenuated in vitro and in vivo, and possibly expressed the foreign antigen on its surface. Therefore, the P gene-deletion strategy may offer a potentially useful and safer approach for attenuating negative-sense RNA viruses which use phosphoprotein as a cofactor for viral replication.

  6. Enhanced immunosurveillance for animal morbilliviruses using vesicular stomatitis virus (VSV) pseudotypes.

    Science.gov (United States)

    Logan, Nicola; Dundon, William G; Diallo, Adama; Baron, Michael D; James Nyarobi, M; Cleaveland, Sarah; Keyyu, Julius; Fyumagwa, Robert; Hosie, Margaret J; Willett, Brian J

    2016-11-11

    The measurement of virus-specific neutralising antibodies represents the "gold-standard" for diagnostic serology. For animal morbilliviruses, such as peste des petits ruminants (PPRV) or rinderpest virus (RPV), live virus-based neutralisation tests require high-level biocontainment to prevent the accidental escape of the infectious agents. In this study, we describe the adaptation of a replication-defective vesicular stomatitis virus (VSVΔG) based pseudotyping system for the measurement of neutralising antibodies against animal morbilliviruses. By expressing the haemagglutinin (H) and fusion (F) proteins of PPRV on VSVΔG pseudotypes bearing a luciferase marker gene, neutralising antibody titres could be measured rapidly and with high sensitivity. Serological responses against the four distinct lineages of PPRV could be measured simultaneously and cross-neutralising responses against other morbilliviruses compared. Using this approach, we observed that titres of neutralising antibodies induced by vaccination with live attenuated PPRV were lower than those induced by wild type virus infection and the level of cross-lineage neutralisation varied between vaccinates. By comparing neutralising responses from animals infected with either PPRV or RPV, we found that responses were highest against the homologous virus, indicating that retrospective analyses of serum samples could be used to confirm the nature of the original pathogen to which an animal had been exposed. Accordingly, when screening sera from domestic livestock and wild ruminants in Tanzania, we detected evidence of cross-species infection with PPRV, canine distemper virus (CDV) and a RPV-related bovine morbillivirus, suggesting that exposure to animal morbilliviruses may be more widespread than indicated previously using existing diagnostic techniques. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

  7. Molecular Determinants of Susceptibility to Oncolytic Vesicular Stomatitis Virus in Pancreatic Adenocarcinoma

    Science.gov (United States)

    Blackham, Aaron U; Northrup, Scott A; Willingham, Mark; Sirintrapun, Joseph; Russell, Greg B; Lyles, Douglas S; Stewart, John H

    2014-01-01

    Background M protein mutant vesicular stomatitis virus (M51R-VSV) has oncolytic properties against many cancers. However, some cancer cells are resistant to M51R-VSV. Herein, we evaluate the molecular determinants of VSV resistance in pancreatic adenocarcinoma cells. Methods Cell viability and the effect of β-interferon (IFN) was analyzed using MTS assay. Gene expression was evaluated via microarray analysis. Cell infectability was measured by flow cytometry. Xenografts were established in athymic nude mice and treated with intratumoral M51R-VSV. Results Four of five pancreatic cancer cell lines were sensitive to M51R-VSV, while Panc03.27 cells remained resistant (81±3% viability 72-hours after single cycle infection). Comparing sensitive MiaPaCa2 to resistant Panc03.27 cells, significant differences in gene expression was found relating to IFN signaling (p=2×10-5), viral entry (p=3×10-4) and endocytosis (p=7×10-4). MiaPaCa2 cells permitted high levels of VSV infection, while Panc03.27 cells were capable of resisting VSV cell entry even at high MOIs. Extrinsic β-IFN overcame apparent defects in IFN-mediated pathways in MiaPaCa2 cells conferring VSV resistance. In contrast, β-IFN decreased cell viability in Panc3.27 cells suggesting intact anti-viral mechanisms. VSV treated xenografts exhibited reduced tumor growth relative to controls in both MiaPaCa2 (1423 ± 345% vs 164 ± 136%, pVSV treated Panc03.27 xenografts. Conclusions Inhibition of VSV endocytosis and intact IFN-mediated defenses are responsible for M51R-VSV resistance in pancreatic adenocarcinoma cells. M51R-VSV treatment appears to induce anti-tumor cellular immunity in vivo which may expand its clinical efficacy. PMID:24252853

  8. Highly attenuated recombinant vesicular stomatitis virus VSV-12'GFP displays immunogenic and oncolytic activity.

    Science.gov (United States)

    van den Pol, Anthony N; Davis, John N

    2013-01-01

    Vesicular stomatitis virus (VSV) has shown considerable promise both as an immunization vector and as an oncolytic virus. In both applications, an important concern is the safety profile of the virus. To generate a highly attenuated virus, we added two reporter genes to the 3' end of the VSV genome, thereby shifting the NPMGL genes from positions 1 to 5 to positions 3 to 7. The resulting virus (VSV-12'GFP) was highly attenuated, generating smaller plaques than four other attenuated VSVs. In one-step growth curves, VSV-12'GFP displayed the slowest growth kinetics. The mechanism of attenuation appears to be due to reduced expression of VSV genes downstream of the reporter genes, as suggested by a 10.4-fold reduction in L-protein RNA transcript. Although attenuated, VSV-12'GFP was highly effective at generating an immune response, indicated by a high-titer antibody response against the green fluorescent protein (GFP) expressed by the virus. Although VSV-12'GFP was more attenuated than other VSVs on both normal and cancer cells, it nonetheless showed a greater level of infection of human cancer cells (glioma and melanoma) than of normal cells, and this effect was magnified in glioma by interferon application, indicating selective oncolysis. Intravenous VSV-12'GFP selectively infected human gliomas implanted into SCID mice subcutaneously or intracranially. All postnatal day 16 mice given intranasal VSV-12'GFP survived, whereas only 10% of those given VSV-G/GFP survived, indicating reduced neurotoxicity. Intratumoral injection of tumors with VSV-12'GFP dramatically suppressed tumor growth and enhanced survival. Together these data suggest this recombinant virus merits further study for its oncolytic and vaccine potential.

  9. In Vitro and In Vivo Attenuation of Vesicular Stomatitis Virus (VSV by Phosphoprotein Deletion.

    Directory of Open Access Journals (Sweden)

    Phonphimon Wongthida

    Full Text Available Vesicular stomatitis virus (VSV is highly immunogenic and able to stimulate both innate and adaptive immune responses. However, its ability to induce adverse effects has held back the use of VSV as a potential vaccine vector. In this study we developed VSV-ΔP, a safe yet potent replication-defective recombinant VSV in which the phosphoprotein (P gene was deleted. VSV-ΔP replicated only in supporting cells expressing P (BHK-P cells and at levels more than 2 logs lower than VSV. In vivo studies indicated that the moderate replication of VSV-ΔP in vitro was associated with the attenuation of this virus in the mouse model, whereas mice intracranially injected with VSV succumbed to neurotoxicity. Furthermore, we constructed VSV and VSV-ΔP expressing a variety of antigens including hemagglutinin-neuraminidase (HN from Newcastle disease virus (NDV, hemagglutinin (HA from either a 2009 H1N1 pandemic influenza virus (pdm/09 or the avian H7N9. VSV and VSV-ΔP incorporated the foreign antigens on their surface resulting in induction of robust neutralizing antibody, serum IgG, and hemagglutination inhibition (HAI titers against their corresponding viruses. These results indicated that VSV with P gene deletion was attenuated in vitro and in vivo, and possibly expressed the foreign antigen on its surface. Therefore, the P gene-deletion strategy may offer a potentially useful and safer approach for attenuating negative-sense RNA viruses which use phosphoprotein as a cofactor for viral replication.

  10. Glycoprotein cytoplasmic domain sequences required for rescue of a vesicular stomatitis virus glycoprotein mutant

    Energy Technology Data Exchange (ETDEWEB)

    Whitt, M.A.; Chong, L.; Rose, J.K. (Yale Univ. School of Medicine, New Haven, CT (USA))

    1989-09-01

    The authors have used transient expression of the wild-type vesicular stomatitis virus (VSV) glycoprotein (G protein) from cloned cDNA to rescue a temperature-sensitive G protein mutant of VSV in cells at the nonpermissive temperature. Using cDNAs encoding G proteins with deletions in the normal 29-amino-acid cytoplasmic domain, they determined that the presence of either the membrane-proximal 9 amino acids or the membrane-distal 12 amino acids was sufficient for rescue of the temperature-sensitive mutant. G proteins with cytoplasmic domains derived from other cellular or viral G proteins did not rescue the mutant, nor did G proteins with one or three amino acids of the normal cytoplasmic domain. Rescue correlated directly with the ability of the G proteins to be incorporated into virus particles. This was shown by analysis of radiolabeled particles separated on sucrose gradients as well as by electron microscopy of rescued virus after immunogold labeling. Quantitation of surface expression showed that all of the mutated G proteins were expressed less efficiently on the cell surface than was wild-type G protein. However, they were able to correct for differences in rescue efficiency resulting from differences in the level of surface expression by reducing wild-type G protein expression to levels equivalent to those observed for the mutated G proteins. The results provide evidence that at least a portion of the cytoplasmic domain is required for efficient assembly of the VSV G protein into virions during virus budding.

  11. Mutations in the glycoprotein of vesicular stomatitis virus affect cytopathogenicity: potential for oncolytic virotherapy.

    Science.gov (United States)

    Janelle, Valérie; Brassard, Frédérick; Lapierre, Pascal; Lamarre, Alain; Poliquin, Laurent

    2011-07-01

    Vesicular stomatitis virus (VSV) has been widely used to characterize cellular processes, viral resistance, and cytopathogenicity. Recently, VSV has also been used for oncolytic virotherapy due to its capacity to selectively lyse tumor cells. Mutants of the matrix (M) protein of VSV have generally been preferred to the wild-type virus for oncolysis because of their ability to induce type I interferon (IFN) despite causing weaker cytopathic effects. However, due to the large variability of tumor types, it is quite clear that various approaches and combinations of multiple oncolytic viruses will be needed to effectively treat most cancers. With this in mind, our work focused on characterizing the cytopathogenic profiles of four replicative envelope glycoprotein (G) VSV mutants. In contrast to the prototypic M mutant, VSV G mutants are as efficient as wild-type virus at inhibiting cellular transcription and host protein translation. Despite being highly cytopathic, the mutant G(6R) triggers type I interferon secretion as efficiently as the M mutant. Importantly, most VSV G mutants are more effective at killing B16 and MC57 tumor cells in vitro than the M mutant or wild-type virus through apoptosis induction. Taken together, our results demonstrate that VSV G mutants retain the high cytopathogenicity of wild-type VSV, with G(6R) inducing type I IFN secretion at levels similar to that of the M mutant. VSV G protein mutants could therefore prove to be highly valuable for the development of novel oncolytic virotherapy strategies that are both safe and efficient for the treatment of various types of cancer.

  12. Resistance of pancreatic cancer cells to oncolytic vesicular stomatitis virus: role of type I interferon signaling.

    Science.gov (United States)

    Moerdyk-Schauwecker, Megan; Shah, Nirav R; Murphy, Andrea M; Hastie, Eric; Mukherjee, Pinku; Grdzelishvili, Valery Z

    2013-02-05

    Oncolytic virus (OV) therapy takes advantage of common cancer characteristics, such as defective type I interferon (IFN) signaling, to preferentially infect and kill cancer cells with viruses. Our recent study (Murphy et al., 2012. J. Virol. 86, 3073-87) found human pancreatic ductal adenocarcinoma (PDA) cells were highly heterogeneous in their permissiveness to vesicular stomatitis virus (VSV) and suggested at least some resistant cell lines retained functional type I IFN responses. Here we examine cellular responses to infection by the oncolytic VSV recombinant VSV-ΔM51-GFP by analyzing a panel of 11 human PDA cell lines for expression of 33 genes associated with type I IFN pathways. Although all cell lines sensed infection by VSV-ΔM51-GFP and most activated IFN-α and β expression, only resistant cell lines displayed constitutive high-level expression of the IFN-stimulated antiviral genes MxA and OAS. Inhibition of JAK/STAT signaling decreased levels of MxA and OAS and increased VSV infection, replication and oncolysis, further implicating IFN responses in resistance. Unlike VSV, vaccinia and herpes simplex virus infectivity and killing of PDA cells was independent of the type I IFN signaling profile, possibly because these two viruses are better equipped to evade type I IFN responses. Our study demonstrates heterogeneity in the type I IFN signaling status of PDA cells and suggests MxA and OAS as potential biomarkers for PDA resistance to VSV and other OVs sensitive to type I IFN responses. Copyright © 2012 Elsevier Inc. All rights reserved.

  13. Interation of mRNA with proteins in vesicular stomatitis virus-infected cells

    Energy Technology Data Exchange (ETDEWEB)

    Adam, S.A.; Choi, Y.D.; Dreyfuss, G.

    1986-02-01

    The interaction of mRNA with proteins in vesicular stomatitis virus (VSV)-infected cells was studied by photochemical cross-linking in intact cells. The major (/sup 35/S)methionine-labeled proteins which became cross-linked by UV light to mRNA in uninfected and in VSV-infected HeLa cells were similar and had apparent mobilities in sodium dodecyl sulfate-polyacrylamide gel electrophoresis corresponding to 135, 93, 72, 68, 53, 50, 43, and 36 kilodaltons. The proteins which were cross-linked in vivo specifically to the five mRNAs of VSV were labeled through radioactive nucleotides incorporated only into VSV mRNAs under conditions in which only VSV mRNAs are labeled. The same major mRNP proteins that became cross-linked to host mRNAs also became cross-linked to VSV mRNAs, although several quantitative differences were detected. Photochemical cross-linking and immunoblotting of cross-linked mRNPs with VSV antiserum demonstrated that in addition to host proteins VSV mRNAs also became cross-linked to the VSV-encoded N protein. The poly(A) segment of both host and VSV mRNAs was associated in vivo selectively with the 72-kilodalton polypeptide. The major proteins of mRNA-ribonucleoprotein complexes are therefore ubiquitous and common to different mRNAs. Furthermore, since the major messenger ribonucleoproteins interact also with VSV mRNAs even though these mRNAs are transcribed in the cytoplasm, it appears that nuclear transcription and nucleocytoplasmic transport are not necessary for mRNA to interact with these proteins.

  14. Cellular proteins associated with the interior and exterior of vesicular stomatitis virus virions.

    Science.gov (United States)

    Moerdyk-Schauwecker, Megan; Hwang, Sun-Il; Grdzelishvili, Valery Z

    2014-01-01

    Virus particles (virions) often contain not only virus-encoded but also host-encoded proteins. Some of these host proteins are enclosed within the virion structure, while others, in the case of enveloped viruses, are embedded in the host-derived membrane. While many of these host protein incorporations are likely accidental, some may play a role in virus infectivity, replication and/or immunoreactivity in the next host. Host protein incorporations may be especially important in therapeutic applications where large numbers of virus particles are administered. Vesicular stomatitis virus (VSV) is the prototypic rhabdovirus and a candidate vaccine, gene therapy and oncolytic vector. Using mass spectrometry, we previously examined cell type dependent host protein content of VSV virions using intact ("whole") virions purified from three cell lines originating from different species. Here we aimed to determine the localization of host proteins within the VSV virions by analyzing: i) whole VSV virions; and ii) whole VSV virions treated with Proteinase K to remove all proteins outside the viral envelope. A total of 257 proteins were identified, with 181 identified in whole virions and 183 identified in Proteinase K treated virions. Most of these proteins have not been previously shown to be associated with VSV. Functional enrichment analysis indicated the most overrepresented categories were proteins associated with vesicles, vesicle-mediated transport and protein localization. Using western blotting, the presence of several host proteins, including some not previously shown in association with VSV (such as Yes1, Prl1 and Ddx3y), was confirmed and their relative quantities in various virion fractions determined. Our study provides a valuable inventory of virion-associated host proteins for further investigation of their roles in the replication cycle, pathogenesis and immunoreactivity of VSV.

  15. Type III interferon attenuates a vesicular stomatitis virus-based vaccine vector.

    Science.gov (United States)

    Guayasamin, Ryann C; Reynolds, Tracy D; Wei, Xin; Fujiwara, Mai; Robek, Michael D

    2014-09-01

    Vesicular stomatitis virus (VSV) has been extensively studied as a vaccine vector and oncolytic agent. Nevertheless, safety concerns have limited its widespread use in humans. The type III lambda interferon (IFN-λ) family of cytokines shares common signaling pathways with the IFN-α/β family and thus evokes similar antiviral activities. However, IFN-λ signals through a distinct receptor complex that is expressed in a cell type-specific manner, which restricts its activity to epithelial barriers, particularly those corresponding to the respiratory and gastrointestinal tracts. In this study, we determined how IFN-λ expression from recombinant VSV would influence vector replication, spread, and immunogenicity. We demonstrate that IFN-λ expression severely attenuates VSV in cell culture. In vivo, IFN-λ limits VSV replication in the mouse lung after intranasal administration and reduces virus spread to other organs. Despite this attenuation, however, the vector retains its capacity to induce protective CD8 T cell and antibody responses after a single immunization. These findings demonstrate a novel method of viral vector attenuation that could be used in both vaccine and oncolytic virus applications. Viruses such as VSV that are used as vaccine vectors can induce protective T cell and antibody responses after a single dose. Additionally, IFN-λ is a potent antiviral agent that has certain advantages for clinical use compared to IFN-α/β, such as fewer patient side effects. Here, we demonstrate that IFN-λ attenuates VSV replication and spread following intranasal virus delivery but does not reduce the ability of VSV to induce potent protective immune responses. These findings demonstrate that the type III IFN family may have widespread applicability for improving the safety and efficacy of viral vaccine and oncolytic vectors. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  16. Induction of Stress Granule-Like Structures in Vesicular Stomatitis Virus-Infected Cells

    Science.gov (United States)

    Dinh, Phat X.; Beura, Lalit K.; Das, Phani B.; Panda, Debasis; Das, Anshuman

    2013-01-01

    Previous studies from our laboratory revealed that cellular poly(C) binding protein 2 (PCBP2) downregulates vesicular stomatitis virus (VSV) gene expression. We show here that VSV infection induces the formation of granular structures in the cytoplasm containing cellular RNA-binding proteins, including PCBP2, T-cell-restricted intracellular antigen 1 (TIA1), and TIA1-related protein (TIAR). Depletion of TIA1 via small interfering RNAs (siRNAs), but not depletion of TIAR, results in enhanced VSV growth and gene expression. The VSV-induced granules appear to be similar to the stress granules (SGs) generated in cells triggered by heat shock or oxidative stress but do not contain some of the bona fide SG markers, such as eukaryotic initiation factor 3 (eIF3) or eIF4A, or the processing body (PB) markers, such as mRNA-decapping enzyme 1A (DCP1a), and thus may not represent canonical SGs or PBs. Our results revealed that the VSV-induced granules, called SG-like structures here, contain the viral replicative proteins and RNAs. The formation and maintenance of the SG-like structures required viral replication and ongoing protein synthesis, but an intact cytoskeletal network was not necessary. These results suggest that cells respond to VSV infection by aggregating the antiviral proteins, such as PCBP2 and TIA1, to form SG-like structures. The functional significance of these SG-like structures in VSV-infected cells is currently under investigation. PMID:23077311

  17. Antagonistic Effects of Cellular Poly(C) Binding Proteins on Vesicular Stomatitis Virus Gene Expression ▿

    Science.gov (United States)

    Dinh, Phat X.; Beura, Lalit K.; Panda, Debasis; Das, Anshuman; Pattnaik, Asit K.

    2011-01-01

    Immunoprecipitation and subsequent mass spectrometry analysis of the cellular proteins from cells expressing the vesicular stomatitis virus (VSV) P protein identified the poly(C) binding protein 2 (PCBP2) as one of the P protein-interacting proteins. To investigate the role of PCBP2 in the viral life cycle, we examined the effects of depletion or overexpression of this protein on VSV growth. Small interfering RNA-mediated silencing of PCBP2 promoted VSV replication. Conversely, overexpression of PCBP2 in transfected cells suppressed VSV growth. Further studies revealed that PCBP2 negatively regulates overall viral mRNA accumulation and subsequent genome replication. Coimmunoprecipitation and immunofluorescence microscopic studies showed that PCBP2 interacts and colocalizes with VSV P protein in virus-infected cells. The P-PCBP2 interaction did not result in reduced levels of protein complex formation with the viral N and L proteins, nor did it induce degradation of the P protein. In addition, PCBP1, another member of the poly(C) binding protein family with homology to PCBP2, was also found to interact with the P protein and inhibit the viral mRNA synthesis at the level of primary transcription without affecting secondary transcription or genome replication. The inhibitory effects of PCBP1 on VSV replication were less pronounced than those of PCBP2. Overall, the results presented here suggest that cellular PCBP2 and PCBP1 antagonize VSV growth by affecting viral gene expression and highlight the importance of these two cellular proteins in restricting virus infections. PMID:21752917

  18. Cellular proteins associated with the interior and exterior of vesicular stomatitis virus virions.

    Directory of Open Access Journals (Sweden)

    Megan Moerdyk-Schauwecker

    Full Text Available Virus particles (virions often contain not only virus-encoded but also host-encoded proteins. Some of these host proteins are enclosed within the virion structure, while others, in the case of enveloped viruses, are embedded in the host-derived membrane. While many of these host protein incorporations are likely accidental, some may play a role in virus infectivity, replication and/or immunoreactivity in the next host. Host protein incorporations may be especially important in therapeutic applications where large numbers of virus particles are administered. Vesicular stomatitis virus (VSV is the prototypic rhabdovirus and a candidate vaccine, gene therapy and oncolytic vector. Using mass spectrometry, we previously examined cell type dependent host protein content of VSV virions using intact ("whole" virions purified from three cell lines originating from different species. Here we aimed to determine the localization of host proteins within the VSV virions by analyzing: i whole VSV virions; and ii whole VSV virions treated with Proteinase K to remove all proteins outside the viral envelope. A total of 257 proteins were identified, with 181 identified in whole virions and 183 identified in Proteinase K treated virions. Most of these proteins have not been previously shown to be associated with VSV. Functional enrichment analysis indicated the most overrepresented categories were proteins associated with vesicles, vesicle-mediated transport and protein localization. Using western blotting, the presence of several host proteins, including some not previously shown in association with VSV (such as Yes1, Prl1 and Ddx3y, was confirmed and their relative quantities in various virion fractions determined. Our study provides a valuable inventory of virion-associated host proteins for further investigation of their roles in the replication cycle, pathogenesis and immunoreactivity of VSV.

  19. Interference of CD40L-mediated tumor immunotherapy by oncolytic vesicular stomatitis virus.

    Science.gov (United States)

    Galivo, Feorillo; Diaz, Rosa Maria; Thanarajasingam, Uma; Jevremovic, Dragan; Wongthida, Phonphimon; Thompson, Jill; Kottke, Timothy; Barber, Glen N; Melcher, Alan; Vile, Richard G

    2010-04-01

    Oncolytic virotherapy can be achieved in two ways: (1) by exploiting an innate ability of certain viruses to selectively replicate in tumor tissues, and (2) by using viruses to deliver toxic or immunostimulatory genes to tumors. Vesicular stomatitis virus (VSV) selectively replicates in tumors lacking adequate type I interferon response. The efficacy of oncolytic virotherapy using VSV against B16 melanomas in C57BL/6 mice is dependent on CD8(+) T and natural killer cells. Because immunotherapies that prime specific CD8(+) T cells against melanocyte/melanoma antigens can generate significant therapeutic responses, we hypothesized that engineering VSV to express the potent T cell costimulatory molecule CD40 ligand (VSV-CD40L) would enhance virotherapy with concomitant priming of melanoma-specific T cells. However, we observed no difference in antitumor efficacy between the parental VSV-GFP and VSV-CD40L. In contrast, intratumoral injection of a replication-defective adenovirus expressing CD40L (Ad-CD40L) consistently produced significantly greater therapy than either replication-competent VSV-GFP or VSV-CD40L. The Ad-CD40L-mediated tumor regressions were associated with specific T cell responses against tumor-associated antigens (TAAs), which took several days to develop, whereas VSV-CD40L rapidly induced high levels of T cell activation without specificity for TAAs. These data suggest that the high levels of VSV-associated immunogenicity distracted immune responses away from priming of tumor-specific T cells, even in the presence of potent costimulatory signals. In contrast, a replication-defective Ad-CD40L allowed significant priming of T cells directed against TAAs. These observations suggest that an efficiently primed antitumor T cell response can produce similar, if not better, therapy against an established melanoma compared with intratumoral injection of a replication-competent oncolytic virus.

  20. Characteristics of oncolytic vesicular stomatitis virus displaying tumor-targeting ligands.

    Science.gov (United States)

    Ammayappan, Arun; Peng, Kah-Whye; Russell, Stephen J

    2013-12-01

    We sought proof of principle that tumor-targeting ligands can be displayed on the surface of vesicular stomatitis virus (VSV) by engineering its glycoprotein. Here, we successfully rescued VSVs displaying tumor vasculature-targeting ligands. By using a rational approach, we investigated various feasible insertion sites on the G protein of VSV (VSV-G) for display of tumor vasculature-targeting ligands, cyclic RGD (cRGD) and echistatin. We found seven sites on VSV-G that tolerated insertion of the 9-residue cRGD peptide, two of which could tolerate insertion of the 49-amino acid echistatin domain. All of the ligand-displaying viruses replicated as well as the parental virus. In vitro studies demonstrated that the VSV-echistatin viruses specifically bound to targeted integrins. Since the low-density lipoprotein receptor (LDLR) was recently identified as a major receptor for VSV, we investigated the entry of ligand-displaying viruses after masking LDLR. The experiment showed that the modified viruses can enter the cell independently of LDLR, whereas entry of unmodified virus is significantly blocked by a specific monoclonal antibody against LDLR. Both parental and ligand-displaying viruses displayed equal oncolytic efficacies in a syngeneic mouse myeloma model. We further demonstrated that single-chain antibody fragments against tumor-specific antigens can be inserted at the N terminus of the G protein and that corresponding replication-competent VSVs can be rescued efficiently. Overall, we demonstrated that functional tumor-targeting ligands can be displayed on replication-competent VSVs without perturbing viral growth and oncolytic efficacy. This study provides a rational foundation for the future development of fully retargeted oncolytic VSVs.

  1. Vesicular stomatitis virus-based ebola vaccine is well-tolerated and protects immunocompromised nonhuman primates.

    Directory of Open Access Journals (Sweden)

    Thomas W Geisbert

    2008-11-01

    Full Text Available Ebola virus (EBOV is a significant human pathogen that presents a public health concern as an emerging/re-emerging virus and as a potential biological weapon. Substantial progress has been made over the last decade in developing candidate preventive vaccines that can protect nonhuman primates against EBOV. Among these prospects, a vaccine based on recombinant vesicular stomatitis virus (VSV is particularly robust, as it can also confer protection when administered as a postexposure treatment. A concern that has been raised regarding the replication-competent VSV vectors that express EBOV glycoproteins is how these vectors would be tolerated by individuals with altered or compromised immune systems such as patients infected with HIV. This is especially important as all EBOV outbreaks to date have occurred in areas of Central and Western Africa with high HIV incidence rates in the population. In order to address this concern, we evaluated the safety of the recombinant VSV vector expressing the Zaire ebolavirus glycoprotein (VSVDeltaG/ZEBOVGP in six rhesus macaques infected with simian-human immunodeficiency virus (SHIV. All six animals showed no evidence of illness associated with the VSVDeltaG/ZEBOVGP vaccine, suggesting that this vaccine may be safe in immunocompromised populations. While one goal of the study was to evaluate the safety of the candidate vaccine platform, it was also of interest to determine if altered immune status would affect vaccine efficacy. The vaccine protected 4 of 6 SHIV-infected macaques from death following ZEBOV challenge. Evaluation of CD4+ T cells in all animals showed that the animals that succumbed to lethal ZEBOV challenge had the lowest CD4+ counts, suggesting that CD4+ T cells may play a role in mediating protection against ZEBOV.

  2. Comparative oncology evaluation of intravenous recombinant oncolytic Vesicular Stomatitis Virus therapy in spontaneous canine cancer.

    Science.gov (United States)

    Naik, Shruthi; Galyon, Gina D; Jenks, Nathan J; Steele, Michael B; Miller, Amber C; Allstadt, Sara D; Suksanpaisan, Lukkana; Peng, Kah Whye; Federspiel, Mark J; Russell, Stephen J; LeBlanc, Amy K

    2017-11-20

    Clinical translation of intravenous therapies to treat disseminated or metastatic cancer is imperative. Comparative oncology, the evaluation of novel cancer therapies in animals with spontaneous cancer, can be utilized to inform and accelerate clinical translation. Preclinical murine studies demonstrate that single shot systemic therapy with a VSV-IFNβ-NIS, a novel recombinant oncolytic Vesicular stomatitis virus (VSV), can induce curative remission in tumor bearing mice. Clinical translation of VSV-IFNβ-NIS therapy is dependent on comprehensive assessment of clinical toxicities, virus shedding, pharmacokinetics, and efficacy in clinically relevant models. Dogs spontaneously develop cancer with comparable etiology, clinical progression and response to therapy as human malignancies. A comparative oncology study was carried out to investigate feasibility and tolerability of intravenous oncolytic VSV-IFNβ-NIS therapy in pet dogs with spontaneous cancer. Nine dogs with various malignancies were treated with a single intravenous dose of VSV-IFNβ-NIS. Two dogs with high-grade peripheral T-cell lymphoma had rapid but transient remission of disseminated disease and transient hepatotoxicity that resolved spontaneously. There was no shedding of infectious virus. Correlative pharmacokinetic studies revealed elevated levels of VSV RNA in blood in dogs with measurable disease remission. This is the first evaluation of intravenous oncolytic virus therapy for spontaneous canine cancer, demonstrating that VSV-IFNβ-NIS is well-tolerated and safe in dogs with advanced or metastatic disease. This approach has informed clinical translation, including dose and target indication selection, leading to a clinical investigation of intravenous VSV-IFNβ-NIS therapy, and provided preliminary evidence of clinical efficacy, and potential biomarkers that correlate with therapeutic response. Copyright ©2017, American Association for Cancer Research.

  3. Semireplication-competent vesicular stomatitis virus as a novel platform for oncolytic virotherapy.

    Science.gov (United States)

    Muik, Alexander; Dold, Catherine; Geiß, Yvonne; Volk, Andreas; Werbizki, Marina; Dietrich, Ursula; von Laer, Dorothee

    2012-08-01

    Among oncolytic viruses, the vesicular stomatitis virus (VSV) is especially potent and a highly promising agent for the treatment of cancer. But, even though effective against multiple tumor entities in preclinical animal models, replication-competent VSV exhibits inherent neurovirulence, which has so far hindered clinical development. To overcome this limitation, replication-defective VSV vectors for cancer gene therapy have been tested and proven to be safe. However, gene delivery was inefficient and only minor antitumor efficacy was observed. Here, we present semireplication-competent vector systems for VSV (srVSV), composed of two trans-complementing, propagation-deficient VSV vectors. The de novo generated deletion mutants of the two VSV polymerase proteins P (phosphoprotein) and L (large catalytic subunit), VSVΔP and VSVΔL respectively, were used mutually or in combination with VSVΔG vectors. These srVSV systems copropagated in vitro and in vivo without recombinatory reversion to replication-competent virus. The srVSV systems were highly lytic for human glioblastoma cell lines, spheroids, and subcutaneous xenografts. Especially the combination of VSVΔG/VSVΔL vectors was as potent as wild-type VSV (VSV-WT) in vitro and induced long-term tumor regression in vivo without any associated adverse effects. In contrast, 90% of VSV-WT-treated animals succumbed to neurological disease shortly after tumor clearance. Most importantly, even when injected into the brain, VSVΔG/VSVΔL did not show any neurotoxicity. In conclusion, srVSV is a promising platform for virotherapeutic approaches and also for VSV-based vector vaccines, combining improved safety with an increased coding capacity for therapeutic transgenes, potentially allowing for multipronged approaches.

  4. Current good manufacturing practice production of an oncolytic recombinant vesicular stomatitis viral vector for cancer treatment.

    Science.gov (United States)

    Ausubel, L J; Meseck, M; Derecho, I; Lopez, P; Knoblauch, C; McMahon, R; Anderson, J; Dunphy, N; Quezada, V; Khan, R; Huang, P; Dang, W; Luo, M; Hsu, D; Woo, S L C; Couture, L

    2011-04-01

    Vesicular stomatitis virus (VSV) is an oncolytic virus currently being investigated as a promising tool to treat cancer because of its ability to selectively replicate in cancer cells. To enhance the oncolytic property of the nonpathologic laboratory strain of VSV, we generated a recombinant vector [rVSV(MΔ51)-M3] expressing murine gammaherpesvirus M3, a secreted viral chemokine-binding protein that binds to a broad range of mammalian chemokines with high affinity. As previously reported, when rVSV(MΔ51)-M3 was used in an orthotopic model of hepatocellular carcinoma (HCC) in rats, it suppressed inflammatory cell migration to the virus-infected tumor site, which allowed for enhanced intratumoral virus replication leading to increased tumor necrosis and substantially prolonged survival. These encouraging results led to the development of this vector for clinical translation in patients with HCC. However, a scalable current Good Manufacturing Practice (cGMP)-compliant manufacturing process has not been described for this vector. To produce the quantities of high-titer virus required for clinical trials, a process that is amenable to GMP manufacturing and scale-up was developed. We describe here a large-scale (50-liter) vector production process capable of achieving crude titers on the order of 10(9) plaque-forming units (PFU)/ml under cGMP. This process was used to generate a master virus seed stock and a clinical lot of the clinical trial agent under cGMP with an infectious viral titer of approximately 2 × 10(10) PFU/ml (total yield, 1 × 10(13) PFU). The lot has passed all U.S. Food and Drug Administration-mandated release testing and will be used in a phase 1 clinical translational trial in patients with advanced HCC.

  5. Reovirus FAST Protein Enhances Vesicular Stomatitis Virus Oncolytic Virotherapy in Primary and Metastatic Tumor Models

    Directory of Open Access Journals (Sweden)

    Fabrice Le Boeuf

    2017-09-01

    Full Text Available The reovirus fusion-associated small transmembrane (FAST proteins are the smallest known viral fusogens (∼100–150 amino acids and efficiently induce cell-cell fusion and syncytium formation in multiple cell types. Syncytium formation enhances cell-cell virus transmission and may also induce immunogenic cell death, a form of apoptosis that stimulates immune recognition of tumor cells. These properties suggest that FAST proteins might serve to enhance oncolytic virotherapy. The oncolytic activity of recombinant VSVΔM51 (an interferon-sensitive vesicular stomatitis virus [VSV] mutant encoding the p14 FAST protein (VSV-p14 was compared with a similar construct encoding GFP (VSV-GFP in cell culture and syngeneic BALB/c tumor models. Compared with VSV-GFP, VSV-p14 exhibited increased oncolytic activity against MCF-7 and 4T1 breast cancer spheroids in culture and reduced primary 4T1 breast tumor growth in vivo. VSV-p14 prolonged survival in both primary and metastatic 4T1 breast cancer models, and in a CT26 metastatic colon cancer model. As with VSV-GFP, VSV-p14 preferentially replicated in vivo in tumors and was cleared rapidly from other sites. Furthermore, VSV-p14 increased the numbers of activated splenic CD4, CD8, natural killer (NK, and natural killer T (NKT cells, and increased the number of activated CD4 and CD8 cells in tumors. FAST proteins may therefore provide a multi-pronged approach to improving oncolytic virotherapy via syncytium formation and enhanced immune stimulation.

  6. Vesicular stomatitis virus-based vaccines protect nonhuman primates against Bundibugyo ebolavirus.

    Directory of Open Access Journals (Sweden)

    Chad E Mire

    Full Text Available Ebola virus (EBOV causes severe and often fatal hemorrhagic fever in humans and nonhuman primates (NHPs. Currently, there are no licensed vaccines or therapeutics for human use. Recombinant vesicular stomatitis virus (rVSV-based vaccine vectors, which encode an EBOV glycoprotein in place of the VSV glycoprotein, have shown 100% efficacy against homologous Sudan ebolavirus (SEBOV or Zaire ebolavirus (ZEBOV challenge in NHPs. In addition, a single injection of a blend of three rVSV vectors completely protected NHPs against challenge with SEBOV, ZEBOV, the former Côte d'Ivoire ebolavirus, and Marburg virus. However, recent studies suggest that complete protection against the newly discovered Bundibugyo ebolavirus (BEBOV using several different heterologous filovirus vaccines is more difficult and presents a new challenge. As BEBOV caused nearly 50% mortality in a recent outbreak any filovirus vaccine advanced for human use must be able to protect against this new species. Here, we evaluated several different strategies against BEBOV using rVSV-based vaccines. Groups of cynomolgus macaques were vaccinated with a single injection of a homologous BEBOV vaccine, a single injection of a blended heterologous vaccine (SEBOV/ZEBOV, or a prime-boost using heterologous SEBOV and ZEBOV vectors. Animals were challenged with BEBOV 29-36 days after initial vaccination. Macaques vaccinated with the homologous BEBOV vaccine or the prime-boost showed no overt signs of illness and survived challenge. In contrast, animals vaccinated with the heterologous blended vaccine and unvaccinated control animals developed severe clinical symptoms consistent with BEBOV infection with 2 of 3 animals in each group succumbing. These data show that complete protection against BEBOV will likely require incorporation of BEBOV glycoprotein into the vaccine or employment of a prime-boost regimen. Fortunately, our results demonstrate that heterologous rVSV-based filovirus vaccine

  7. Oncolytic efficacy of recombinant vesicular stomatitis virus and myxoma virus in experimental models of rhabdoid tumors.

    Science.gov (United States)

    Wu, Yushui; Lun, Xueqing; Zhou, Hongyuan; Wang, Limei; Sun, Beichen; Bell, John C; Barrett, John W; McFadden, Grant; Biegel, Jaclyn A; Senger, Donna L; Forsyth, Peter A

    2008-02-15

    Rhabdoid tumors are highly aggressive pediatric tumors that are usually refractory to available treatments. The purpose of this study was to evaluate the therapeutic potential of two oncolytic viruses, myxoma virus (MV) and an attenuated vesicular stomatitis virus (VSV(DeltaM51)), in experimental models of human rhabdoid tumor. Four human rhabdoid tumor cell lines were cultured in vitro and treated with live or inactivated control virus. Cytopathic effect, viral gene expression, infectious viral titers, and cell viability were examined at various time points after infection. To study viral oncolysis in vivo, human rhabdoid tumor cells were implanted s.c. in the hind flank or intracranially in CD-1 nude mice and treated with intratumoral (i.t.) or i.v. injections of live or UV-inactivated virus. Viral distribution and effects on tumor size and survival were assessed. All rhabdoid tumor cell lines tested in vitro were susceptible to productive lethal infections by MV and VSV(DeltaM51). I.t. injection of live MV or VSV(DeltaM51) dramatically reduced the size of s.c. rhabdoid tumor xenografts compared with control animals. I.v. administration of VSV(DeltaM51) or i.t. injection of MV prolonged the median survival of mice with brain xenografts compared with controls (VSV(DeltaM51): 25 days versus 21 days, log-rank test, P = 0.0036; MV: median survival not reached versus 21 days, log-rank test, P = 0.0007). Most of the MV-treated animals (4 of 6; 66.7%) were alive and apparently "cured" when the experiment was arbitrarily ended (>180 days). These results suggest that VSV(DeltaM51) and MV could be novel effective therapies against human rhabdoid tumor.

  8. Cooperation of B cells and T cells is required for survival of mice infected with vesicular stomatitis virus

    DEFF Research Database (Denmark)

    Thomsen, Allan Randrup; Nansen, A; Andersen, C

    1997-01-01

    To define the role of T cells and B cells in resistance to vesicular stomatitis virus (VSV) infection, knockout mice with different specific immune defects on an identical background were infected i.v. and the outcome of infection was compared; in this way a more complete picture of the relative...... antibodies are pivotal for survival in the early phase of VSV infection, T cells are required for long-term survival, with CD4+ T cells being more effective in controlling this infection than CD8+ T cells....

  9. Quantitative multiplex assay for simultaneous detection and identification of Indiana and New Jersey serotypes of vesicular stomatitis virus

    DEFF Research Database (Denmark)

    Rasmussen, Thomas Bruun; Uttenthal, Åse; Fernandez, Jovita

    2005-01-01

    In order to establish a rapid and reliable system for the detection of vesicular stomatitis virus (VSV), we developed a quantitative reverse transcription-PCR assay for the detection, quantification, and differentiation of the major serotypes, VSV Indiana and VSV New Jersey, using a closed......-tube multiplex format. The detection system is based on the recently invented primer-probe energy transfer (PriProET) system. A region of the gene encoding the RNA-dependent RNA polymerase was amplified by using VSV-specific primers in the presence of two serotype-specific fluorescent probes. By incorporating...... identification of VSV....

  10. Large-population passages of vesicular stomatitis virus in interferon-treated cells select variants of only limited resistance.

    OpenAIRE

    Novella, I S; Cilnis, M; Elena, S F; Kohn, J.; Moya, A; Domingo, E; Holland, J J

    1996-01-01

    Vesicular stomatitis virus (VSV) populations were repeatedly passaged in L-929 cells treated with alpha interferon (IFN-alpha) at levels of 25 U/ml. This IFN-alpha concentration induced a 99.9% inhibition of viral yield in standard infections. Analysis of viral fitness (overall replicative ability measured in direct competition with a reference wild-type VSV) after 21 passages in IFN-treated cells showed only a limited increase or no increase in fitness, compared with the greater increase upo...

  11. The lipidomes of vesicular stomatitis virus, semliki forest virus, and the host plasma membrane analyzed by quantitative shotgun mass spectrometry

    DEFF Research Database (Denmark)

    Kalvodova, Lucie; Sampaio, Julio L; Cordo, Sandra

    2009-01-01

    Although enveloped virus assembly in the host cell is a crucial step in the virus life cycle, it remains poorly understood. One issue is how viruses include lipids in their membranes during budding from infected host cells. To analyze this issue, we took advantage of the fact that baby hamster...... kidney cells can be infected by two different viruses, namely, vesicular stomatitis virus and Semliki Forest virus, from the Rhabdoviridae and Togaviridae families, respectively. We purified the host plasma membrane and the two different viruses after exit from the host cells and analyzed the lipid...

  12. Trojan horse lymphocytes: a vesicular stomatitis virus-specific T-cell clone lyses target cells by carrying virus.

    OpenAIRE

    Hom, R C; Soman, G; Finberg, R

    1989-01-01

    We have isolated a vesicular stomatitis virus (VSV)-specific CD4+ CD8- murine T-cell clone. This clone proliferates only in response to VSV and lyses infected tumor cells bearing class II major histocompatibility antigens in short-term chromium release assays. In addition, the cell has VSV antigens on its surface and is capable of killing uninfected tumor cells without major histocompatibility antigen restriction in a 2-day assay. This latter cytolytic activity is eliminated by anti-VSV antib...

  13. Analysis of virion associated host proteins in vesicular stomatitis virus using a proteomics approach

    Directory of Open Access Journals (Sweden)

    Hwang Sun-Il

    2009-10-01

    Full Text Available Abstract Background Vesicular stomatitis virus (VSV is the prototypic rhabdovirus and the best studied member of the order Mononegavirales. There is now compelling evidence that enveloped virions released from infected cells carry numerous host (cellular proteins some of which may play an important role in viral replication. Although several cellular proteins have been previously shown to be incorporated into VSV virions, no systematic study has been done to reveal the host protein composition for virions of VSV or any other member of Mononegavirales. Results Here we used a proteomics approach to identify cellular proteins within purified VSV virions, thereby creating a "snapshot" of one stage of virus/host interaction that can guide future experiments aimed at understanding molecular mechanisms of virus-cell interactions. Highly purified preparations of VSV virions from three different cell lines of human, mouse and hamster origin were analyzed for the presence of cellular proteins using mass spectrometry. We have successfully confirmed the presence of several previously-identified cellular proteins within VSV virions and identified a number of additional proteins likely to also be present within the virions. In total, sixty-four cellular proteins were identified, of which nine were found in multiple preparations. A combination of immunoblotting and proteinase K protection assay was used to verify the presence of several of these proteins (integrin β1, heat shock protein 90 kDa, heat shock cognate 71 kDa protein, annexin 2, elongation factor 1a within the virions. Conclusion This is, to our knowledge, the first systematic study of the host protein composition for virions of VSV or any other member of the order Mononegavirales. Future experiments are needed to determine which of the identified proteins have an interaction with VSV and whether these interactions are beneficial, neutral or antiviral with respect to VSV replication. Identification

  14. Effects of microwave exposure on the hamster immune system. III. Macrophage resistance to vesicular stomatitis virus infection

    Energy Technology Data Exchange (ETDEWEB)

    Rao, G.R.; Cain, C.A.; Tompkins, W.A.

    1984-01-01

    Exposure of hamsters to microwave (MW) energy (2.45 GHz, 25 mW/cm2, 1 h) resulted in activation of peritoneal macrophages (PM) to a viricidal state restricting the replication of vesicular stomatitis virus (VSV). The PM from MW-exposed hamsters were viricidal as early as 1 day after exposure and remained active for 5 days. Immunization of hamsters with vaccinia virus induced viricidal PM by 3 to 4 days and they remained active for 7 days. To test the hypothesis that thermogenic MW exposure results in the release of endotoxin across the intestinal epithelium which subsequently activates PM, hamsters were injected with lipopolysaccharide (LPS) and their viricidal activity was studied. Lipopolysaccharide in vitro (0.2 microgram) and in vivo (0.5 microgram) activated macrophages to a viricidal state. When administered in vivo, LPS (0.5 microgram) activated macrophages as early as 1 day and the activity remained for 3 days. While MW exposure of PM in vitro failed to induce viricidal activity, exposure of PM to LPS in vitro induced strong viricidal activity. This suggests that the in vivo response of PM to MW is an indirect one, which is consistent with the hypothesis that MW-induced PM viricidal activity may be mediated via LPS. In preliminary experiments, MW exposure resulted in extended survival time for hamsters challenged with a lethal dose of vesicular stomatitis virus, supporting the concept that MW-activated PM may be a useful therapeutic modality.

  15. Inhibition of various steps in the replication cycle of vesicular stomatitis virus contributes to its photoinactivation by AlPcS4 or Pc4 and red light

    NARCIS (Netherlands)

    Moor, ACE; Wagenaars-van Gompel, AE; Hermanns, RCA; van der Meulen, J; Smit, J; Wilschut, J; Brand, A; Dubbelman, TMAR; VanSteveninck, J

    Vesicular stomatitis virus (VSV) was used as a model virus to study the processes involved in photoinactivation by aluminum phthalocyanine tetrasulfonate (AIPcS(4),) or silicon phthalocyanine HOSiPcOSi(CH3)(2)(CH2)(3)N(CH3)(2) (Pc4) and red light. Previously a very rapid decrease in the

  16. Enhancement of immunostimulatory properties of exosomal vaccines by incorporation of fusion-competent G protein of vesicular stomatitis virus.

    Science.gov (United States)

    Temchura, Vladimir V; Tenbusch, Matthias; Nchinda, Godwin; Nabi, Ghulam; Tippler, Bettina; Zelenyuk, Maryna; Wildner, Oliver; Uberla, Klaus; Kuate, Seraphin

    2008-07-04

    Exosomes have been proposed as candidates for therapeutic immunization. The present study demonstrates that incorporation of the G protein of vesicular stomatitis virus (VSV-G) into exosome-like vesicles (ELVs) enhances their uptake and induces the maturation of dendritic cells. Targeting of VSV-G and ovalbumin as a model antigen to the same ELVs increased the cross-presentation of ovalbumin via an endosomal acidification mechanism. Immunization of mice with VSV-G and ovalbumin containing ELVs led to an increased IgG2a antibody response, expansion of antigen-specific CD8 T cells, strong in vivo CTL responses, and protection from challenge with ovalbumin expressing tumor cells. Thus, incorporation of VSV-G and targeting of antigens to ELVs are attractive strategies to improve exosomal vaccines.

  17. Trojan horse lymphocytes: a vesicular stomatitis virus-specific T-cell clone lyses target cells by carrying virus.

    Science.gov (United States)

    Hom, R C; Soman, G; Finberg, R

    1989-10-01

    We have isolated a vesicular stomatitis virus (VSV)-specific CD4+ CD8- murine T-cell clone. This clone proliferates only in response to VSV and lyses infected tumor cells bearing class II major histocompatibility antigens in short-term chromium release assays. In addition, the cell has VSV antigens on its surface and is capable of killing uninfected tumor cells without major histocompatibility antigen restriction in a 2-day assay. This latter cytolytic activity is eliminated by anti-VSV antibody, indicating that its lytic activity is provided by the virus. [35S]methionine labeling and immunoprecipitation experiments demonstrated that viral protein translation is initiated after incubation of the clone with a tumor target cell, defining this as the mechanism of its cytolytic activity.

  18. Strains of Lentinula edodes suppress growth of phytopathogenic fungi and inhibit Alagoas serotype of vesicular stomatitis virus Linhagens de Lentinula edodes inibem fungos fitopatogênicos e o vírus da estomatite vesicular, sorotipo Alagoas

    OpenAIRE

    Sasaki, Selma H.; Rosa E.C. Linhares; Nozawa,Carlos M.; Ricardo Montalván; Paccola-Meirelles,Luzia D.

    2001-01-01

    Four Lentinula edodes strains (Le10, 46, K2, Assai) were assessed for their antagonistic effect on four filamentous fungus species of agricultural importance (Helminthosporium euphorbiae, Helminthosporium sp, Fusarium solani and Phomopsis sojae) and on Alagoas serotype of Vesicular Stomatitis Virus (VSA). The L. edodes strains studied had variable effects on the filamentous fungi and on VSA. The K2 and Le10 strains were antagonistic on the fungi assessed and the 46 and K2 strains were efficie...

  19. Cytopathogenesis of vesicular stomatitis virus is regulated by the PSAP motif of M protein in a species-dependent manner.

    Science.gov (United States)

    Irie, Takashi; Liu, Yuliang; Drolet, Barbara S; Carnero, Elena; García-Sastre, Adolfo; Harty, Ronald N

    2012-09-01

    Vesicular stomatitis virus (VSV) is an important vector-borne pathogen of bovine and equine species, causing a reportable vesicular disease. The matrix (M) protein of VSV is multifunctional and plays a key role in cytopathogenesis, apoptosis, host protein shut-off, and virion assembly/budding. Our previous findings indicated that mutations of residues flanking the (37)PSAP(40) motif within the M protein resulted in VSV recombinants having attenuated phenotypes in mice. In this report, we characterize the phenotype of VSV recombinant PS > A4 (which harbors four alanines (AAAA) in place of the PSAP motif without disruption of flanking residues) in both mice, and in Aedes albopictus C6/36 mosquito and Culicoides sonorensis KC cell lines. The PS > A4 recombinant displayed an attenuated phenotype in infected mice as judged by weight loss, mortality, and viral titers measured from lung and brain samples of infected animals. However, unexpectedly, the PS > A4 recombinant displayed a robust cytopathic phenotype in insect C6/36 cells compared to that observed with control viruses. Notably, titers of recombinant PS > A4 were approximately 10-fold greater than those of control viruses in infected C6/36 cells and in KC cells from Culicoides sonorensis, a known VSV vector species. In addition, recombinant PS > A4 induced a 25-fold increase in the level of C3 caspase activity in infected C6/36 cells. These findings indicate that the PSAP motif plays a direct role in regulating cytopathogenicity in a species-dependent manner, and suggest that the intact PSAP motif may be important for maintaining persistence of VSV in an insect host.

  20. Retargeting Oncolytic Vesicular Stomatitis Virus to Human T-Cell Lymphotropic Virus Type 1-Associated Adult T-Cell Leukemia

    Science.gov (United States)

    Betancourt, Dillon; Ramos, Juan Carlos

    2015-01-01

    ABSTRACT Adult T cell leukemia/lymphoma (ATL) is an aggressive cancer of CD4/CD25+ T lymphocytes, the etiological agent of which is human T-cell lymphotropic virus type 1 (HTLV-1). ATL is highly refractory to current therapies, making the development of new treatments a high priority. Oncolytic viruses such as vesicular stomatitis virus (VSV) are being considered as anticancer agents since they readily infect transformed cells compared to normal cells, the former appearing to exhibit defective innate immune responses. Here, we have evaluated the efficacy and safety of a recombinant VSV that has been retargeted to specifically infect and replicate in transformed CD4+ cells. This was achieved by replacing the single VSV glycoprotein (G) with human immunodeficiency virus type 1 (HIV-1) gp160 to create a hybrid fusion protein, gp160G. The resultant virus, VSV-gp160G, was found to only target cells expressing CD4 and retained robust oncolytic activity against HTLV-1 actuated ATL cells. VSV-gp160G was further noted to be highly attenuated and did not replicate efficiently in or induce significant cell death of primary CD4+ T cells. Accordingly, VSV-gp160G did not elicit any evidence of neurotoxicity even in severely immunocompromised animals such as NOD/Shi-scid, IL-2Rγ-c-null (NSG) mice. Importantly, VSV-gp160G effectively exerted potent oncolytic activity in patient-derived ATL transplanted into NSG mice and facilitated a significant survival benefit. Our data indicate that VSV-gp160G exerts potent oncolytic efficacy against CD4+ malignant cells and either alone or in conjunction with established therapies may provide an effective treatment in patients displaying ATL. IMPORTANCE Adult T cell leukemia (ATL) is a serious form of cancer with a high mortality rate. HTLV-1 infection is the etiological agent of ATL and, unfortunately, most patients succumb to the disease within a few years. Current treatment options have failed to significantly improve survival rate. In

  1. Retargeting Oncolytic Vesicular Stomatitis Virus to Human T-Cell Lymphotropic Virus Type 1-Associated Adult T-Cell Leukemia.

    Science.gov (United States)

    Betancourt, Dillon; Ramos, Juan Carlos; Barber, Glen N

    2015-12-01

    Adult T cell leukemia/lymphoma (ATL) is an aggressive cancer of CD4/CD25(+) T lymphocytes, the etiological agent of which is human T-cell lymphotropic virus type 1 (HTLV-1). ATL is highly refractory to current therapies, making the development of new treatments a high priority. Oncolytic viruses such as vesicular stomatitis virus (VSV) are being considered as anticancer agents since they readily infect transformed cells compared to normal cells, the former appearing to exhibit defective innate immune responses. Here, we have evaluated the efficacy and safety of a recombinant VSV that has been retargeted to specifically infect and replicate in transformed CD4(+) cells. This was achieved by replacing the single VSV glycoprotein (G) with human immunodeficiency virus type 1 (HIV-1) gp160 to create a hybrid fusion protein, gp160G. The resultant virus, VSV-gp160G, was found to only target cells expressing CD4 and retained robust oncolytic activity against HTLV-1 actuated ATL cells. VSV-gp160G was further noted to be highly attenuated and did not replicate efficiently in or induce significant cell death of primary CD4(+) T cells. Accordingly, VSV-gp160G did not elicit any evidence of neurotoxicity even in severely immunocompromised animals such as NOD/Shi-scid, IL-2Rγ-c-null (NSG) mice. Importantly, VSV-gp160G effectively exerted potent oncolytic activity in patient-derived ATL transplanted into NSG mice and facilitated a significant survival benefit. Our data indicate that VSV-gp160G exerts potent oncolytic efficacy against CD4(+) malignant cells and either alone or in conjunction with established therapies may provide an effective treatment in patients displaying ATL. Adult T cell leukemia (ATL) is a serious form of cancer with a high mortality rate. HTLV-1 infection is the etiological agent of ATL and, unfortunately, most patients succumb to the disease within a few years. Current treatment options have failed to significantly improve survival rate. In this study, we

  2. Ruxolitinib and Polycation Combination Treatment Overcomes Multiple Mechanisms of Resistance of Pancreatic Cancer Cells to Oncolytic Vesicular Stomatitis Virus.

    Science.gov (United States)

    Felt, Sébastien A; Droby, Gaith N; Grdzelishvili, Valery Z

    2017-08-15

    Vesicular stomatitis virus (VSV) is a promising oncolytic virus (OV). Although VSV is effective against a majority of pancreatic ductal adenocarcinoma cell (PDAC) cell lines, some PDAC cell lines are highly resistant to VSV, and the mechanisms of resistance are still unclear. JAK1/2 inhibitors (such as ruxolitinib and JAK inhibitor I) strongly stimulate VSV replication and oncolysis in all resistant cell lines but only partially improve the susceptibility of resistant PDACs to VSV. VSV tumor tropism is generally dependent on the permissiveness of malignant cells to viral replication rather than on receptor specificity, with several ubiquitously expressed cell surface molecules playing a role in VSV attachment to host cells. However, as VSV attachment to PDAC cells has never been tested before, here we examined if it was possibly inhibited in resistant PDAC cells. Our data show a dramatically weaker attachment of VSV to HPAF-II cells, the most resistant human PDAC cell line. Although sequence analysis of low-density lipoprotein (LDL) receptor (LDLR) mRNA did not reveal any amino acid substitutions in this cell line, HPAF-II cells displayed the lowest level of LDLR expression and dramatically lower LDL uptake. Treatment of cells with various statins strongly increased LDLR expression levels but did not improve VSV attachment or LDL uptake in HPAF-II cells. However, LDLR-independent attachment of VSV to HPAF-II cells was dramatically improved by treating cells with Polybrene or DEAE-dextran. Moreover, combining VSV with ruxolitinib and Polybrene or DEAE-dextran successfully broke the resistance of HPAF-II cells to VSV by simultaneously improving VSV attachment and replication.IMPORTANCE Oncolytic virus (OV) therapy is an anticancer approach that uses viruses that selectively infect and kill cancer cells. This study focuses on oncolytic vesicular stomatitis virus (VSV) against pancreatic ductal adenocarcinoma (PDAC) cells. Although VSV is effective against most PDAC

  3. Interferon-induced protein Ifit2 protects mice from infection of the peripheral nervous system by vesicular stomatitis virus.

    Science.gov (United States)

    Fensterl, Volker; Wetzel, Jaime L; Sen, Ganes C

    2014-09-01

    The interferon system provides the first line of host defense against virus infection. Mouse pathogenesis studies have revealed the importance of specific interferon-induced proteins in providing protection against specific viruses. We have previously reported that one such protein, Ifit2, protects neurons of the central nervous system from intranasal infection by the neurotropic rhabdovirus, vesicular stomatitis virus (VSV). Here, we demonstrate that Ifit2 protects the peripheral nervous system from VSV infection as well. In Ifit2(-/-) mice, VSV, injected subcutaneously into the footpad, entered the proximal lymph node, where it replicated and infected the nodal nerve endings. The infection spread to the sciatic nerve, the spinal cord, and the brain, causing paralysis. In contrast, in the wild-type mice, although VSV replicated equally well in the lymph node, infection of the sciatic nerve and the rest of the nervous system was impaired, thus preventing paralysis. Ifit2 protected only the nervous system from VSV infection; other tissues were well protected even in Ifit2(-/-) mice. These results indicate that Ifit2 is the interferon-induced protein that prevents VSV infection of neurons of both the peripheral and the central nervous systems, thus inhibiting the consequent neuropathy, but it is dispensable for protecting the cells of other tissues from VSV infection. Although viral infection is quite common, the immune system effectively protects us from viral diseases. A major part of this protection is mediated by interferon, the antiviral cytokine secreted by virus-infected cells. To empower the neighboring uninfected cells in combating the oncoming infection, interferon induces the synthesis of more than 200 new proteins, many of which have antiviral activities. The virus studied here, vesicular stomatitis virus (VSV), like its relative, rabies virus, can cause neuropathy in mice if it enters the peripheral nervous system through skin lesions; however

  4. Recoding of the vesicular stomatitis virus L gene by computer-aided design provides a live, attenuated vaccine candidate.

    Science.gov (United States)

    Wang, Bingyin; Yang, Chen; Tekes, Gergely; Mueller, Steffen; Paul, Aniko; Whelan, Sean P J; Wimmer, Eckard

    2015-03-31

    Codon pair bias (CPB), which has been observed in all organisms, is a neglected genomic phenomenon that affects gene expression. CPB results from synonymous codons that are paired more or less frequently in ORFeomes regardless of codon bias. The effect of an individual codon pair change is usually small, but when it is amplified by large-scale genome recoding, strikingly altered biological phenotypes are observed. The utility of codon pair bias in the development of live attenuated vaccines was recently demonstrated by recodings of poliovirus (a positive-strand RNA virus) and influenza virus (a negative-strand segmented RNA virus). Here, the L gene of vesicular stomatitis virus (VSV), a nonsegmented negative-sense RNA virus, was partially recoded based on codon pair bias. Totals of 858 and 623 silent mutations were introduced into a 5'-terminal segment of the viral L gene (designated L1) to create sequences containing either overrepresented or underrepresented codon pairs, designated L1(sdmax) and L1(min), respectively. Analysis revealed that recombinant VSV containing the L1(min) sequence could not be recovered, whereas the virus with the sdmax sequence showed a modest level of attenuation in cell culture. More strikingly, in mice the L1(sdmax) virus was almost as immunogenic as the parental strain but highly attenuated. Taken together, these results open a new road to attain a balance between VSV virulence and immunogenicity, which could serve as an example for the attenuation of other negative-strand, nonsegmented RNA viruses. Vesicular stomatitis virus (VSV) is the prototypic rhabdovirus in the order Mononegavirales. A wide range of human pathogens belong to this family. Using a unique computer algorithm and large-scale genome synthesis, we attempted to develop a live attenuated vaccine strain for VSV, which could be used as an antigen delivery platform for humans. Recombinant VSVs with distinct codon pair biases were rationally designed, constructed, and

  5. Single-dose, therapeutic vaccination of mice with vesicular stomatitis virus expressing human papillomavirus type 16 E7 protein.

    Science.gov (United States)

    Liao, John B; Publicover, Jean; Rose, John K; DiMaio, Daniel

    2008-05-01

    We are developing recombinant attenuated vesicular stomatitis virus (VSV) as a vaccine vector to generate humoral and cell-mediated immune responses. Here, we explore the use of VSV vaccines for cancer immunotherapy. Immunotherapy targeting high-risk human papillomavirus (HPV) lesions has the potential to benefit HPV-infected individuals and cervical cancer patients by generating cytotoxic T cells that kill tumor cells that express viral antigens. A single dose of VSV expressing the HPV type 16 (HPV16) E7 oncogene was used for therapeutic vaccination of mice bearing TC-1 syngeneic tumors, which express HPV16 E7. HPV16 E7-specific T cells were generated and displayed cytotoxic activity against the tumor cells. By 14 days postvaccination, average tumor volumes were 10-fold less in the vaccinated group than in mice that received the empty-vector VSV, and regression of preexisting tumors occurred in some cases. This antitumor effect was CD8 T-cell dependent. Our results demonstrate antitumor responses to HPV16 E7 and suggest that recombinant-VSV-based vaccination should be explored as a therapeutic strategy for cervical carcinoma and other HPV-associated cancers.

  6. Attenuation of vesicular stomatitis virus infection of brain using antiviral drugs and an adeno-associated virus-interferon vector

    Science.gov (United States)

    Wollmann, Guido; Paglino, Justin C.; Maloney, Patrick R; Ahmadi, Sebastian A; van den Pol, Anthony N

    2015-01-01

    Vesicular stomatitis virus (VSV) shows promise as vaccine-vector and oncolytic virus. However, reports of neurotoxicity of VSV remain a concern. We compared 12 antiviral compounds to control infection of VSV-CT9-M51 and VSV-rp30 using murine and human brain cultures, and in vivo mouse models. Inhibition of replication, cytotoxicity and infectivity was strongest with ribavirin and IFN-α and to some extent with mycophenolic acid, chloroquine, and adenine 9-β-D-arabinofuranoside. To generate continuous IFN exposure, we made an adeno-associated virus vector expressing murine IFN; AAV-mIFN-β protected mouse brain cells from VSV, as did a combination of ribavirin and chloroquine. Intracranial AAV-mIFN-β protected the brain against VSV-CT9-M51. In SCID mice bearing human glioblastoma, AAV-mIFN-β moderately enhanced survival. VSV-CT9-M51 doubled median survival when administered after AAV-mIFN-β; some surviving mice showed complete tumor destruction. Together, these data suggest that AAV-IFN or IFN with ribavirin and chloroquine provide an optimal anti-virus combination against VSV in the brain. PMID:25462341

  7. Oncolytic Vesicular Stomatitis Virus as a Viro-Immunotherapy: Defeating Cancer with a “Hammer” and “Anvil”

    Directory of Open Access Journals (Sweden)

    Michael Karl Melzer

    2017-02-01

    Full Text Available Oncolytic viruses have gained much attention in recent years, due, not only to their ability to selectively replicate in and lyse tumor cells, but to their potential to stimulate antitumor immune responses directed against the tumor. Vesicular stomatitis virus (VSV, a negative-strand RNA virus, is under intense development as an oncolytic virus due to a variety of favorable properties, including its rapid replication kinetics, inherent tumor specificity, and its potential to elicit a broad range of immunomodulatory responses to break immune tolerance in the tumor microenvironment. Based on this powerful platform, a multitude of strategies have been applied to further improve the immune-stimulating potential of VSV and synergize these responses with the direct oncolytic effect. These strategies include: 1. modification of endogenous virus genes to stimulate interferon induction; 2. virus-mediated expression of cytokines or immune-stimulatory molecules to enhance anti-tumor immune responses; 3. vaccination approaches to stimulate adaptive immune responses against a tumor antigen; 4. combination with adoptive immune cell therapy for potentially synergistic therapeutic responses. A summary of these approaches will be presented in this review.

  8. Efficient generation of vesicular stomatitis virus (VSV)-pseudotypes bearing morbilliviral glycoproteins and their use in quantifying virus neutralising antibodies.

    Science.gov (United States)

    Logan, Nicola; McMonagle, Elizabeth; Drew, Angharad A; Takahashi, Emi; McDonald, Michael; Baron, Michael D; Gilbert, Martin; Cleaveland, Sarah; Haydon, Daniel T; Hosie, Margaret J; Willett, Brian J

    2016-02-03

    Morbillivirus neutralising antibodies are traditionally measured using either plaque reduction neutralisation tests (PRNTs) or live virus microneutralisation tests (micro-NTs). While both test formats provide a reliable assessment of the strength and specificity of the humoral response, they are restricted by the limited number of viral strains that can be studied and often present significant biological safety concerns to the operator. In this study, we describe the adaptation of a replication-defective vesicular stomatitis virus (VSVΔG) based pseudotyping system for the measurement of morbillivirus neutralising antibodies. By expressing the haemagglutinin (H) and fusion (F) proteins of canine distemper virus (CDV) on VSVΔG pseudotypes bearing a luciferase marker gene, neutralising antibody titres could be measured rapidly and with high sensitivity. Further, by exchanging the glycoprotein expression construct, responses against distinct viral strains or species may be measured. Using this technique, we demonstrate cross neutralisation between CDV and peste des petits ruminants virus (PPRV). As an example of the value of the technique, we demonstrate that UK dogs vary in the breadth of immunity induced by CDV vaccination; in some dogs the neutralising response is CDV-specific while, in others, the neutralising response extends to the ruminant morbillivirus PPRV. This technique will facilitate a comprehensive comparison of cross-neutralisation to be conducted across the morbilliviruses. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

  9. Oncolytic Vesicular Stomatitis Virus in an Immunocompetent Model of MUC1-Positive or MUC1-Null Pancreatic Ductal Adenocarcinoma

    Science.gov (United States)

    Hastie, Eric; Besmer, Dahlia M.; Shah, Nirav R.; Murphy, Andrea M.; Moerdyk-Schauwecker, Megan; Molestina, Carlos; Roy, Lopamudra Das; Curry, Jennifer M.; Mukherjee, Pinku

    2013-01-01

    Vesicular stomatitis virus (VSV) is a promising oncolytic agent against various malignancies. Here, for the first time, we tested VSV in vitro and in vivo in a clinically relevant, immunocompetent mouse model of pancreatic ductal adenocarcinoma (PDA). Our system allows the study of virotherapy against PDA in the context of overexpression (80% of PDA patients) or no expression of human mucin 1 (MUC1), a major marker for poor prognosis in patients. In vitro, we tested three VSV recombinants, wild-type VSV, VSV-green fluorescent protein (VSV-GFP), and a safe oncolytic VSV-ΔM51-GFP, against five mouse PDA cell lines that either expressed human MUC1 or were MUC1 null. All viruses demonstrated significant oncolytic abilities independent of MUC1 expression, although VSV-ΔM51-GFP was somewhat less effective in two PDA cell lines. In vivo administration of VSV-ΔM51-GFP resulted in significant reduction of tumor growth for tested mouse PDA xenografts (+MUC1 or MUC1 null), and antitumor efficacy was further improved when the virus was combined with the chemotherapeutic drug gemcitabine. The antitumor effect was transient in all tested groups. The developed system can be used to study therapies involving various oncolytic viruses and chemotherapeutics, with the goal of inducing tumor-specific immunity while preventing premature virus clearance. PMID:23864625

  10. Preparation of vesicular stomatitis virus-G (VSV-G) conjugate and its use in gene transfer.

    Science.gov (United States)

    Miyanohara, Atsushi

    2012-04-01

    The fusiogenic envelope G glycoprotein of the vesicular stomatitis virus (VSV-G) that has been used to pseudotype retrovirus and lentivirus vectors can be used alone as an efficient vehicle for gene transfer. VSV-G protein is secreted into the culture medium as sendimentable vesicles from cells transfected with a VSV-G expression plasmid in the absence of other viral components. The VSV-G vesicles in the conditioned medium can be partially purified by pelleting through sucrose cushion ultracentrifugation. Protein-DNA complexes are formed by mixing the VSV-G vesicles with naked plasmid DNA. Such complexes show markedly enhanced transfection efficiency when added to the culture medium of recipient cells. The cell tropism of VSV-G-DNA complex-mediated gene transfer resembles that of VSV-G-pseudotyped retrovirus and lentivirus vectors, and the complex is therefore particularly useful for transfection of cells that are refractory to other methods. Still, some cells are refractory to VSV-G-mediated transfection. It should also be noted that overdose of VSV-G can be quite toxic to the recipient cells. The primitive complexes formed by mixing a viral fusiogenic envelope protein with naked DNA may represent a step toward fusing useful features of viral and nonviral vectors for safer and more efficient gene transfer. This protocol describes simple methods for preparation of VSV-G and for gene transfer with DNA-VSV-G complexes.

  11. Mediators of protection against lethal systemic vesicular stomatitis virus infection in hamsters: defective interfering particles, polyinosinate-polycytidylate, and interferon

    Energy Technology Data Exchange (ETDEWEB)

    Fultz, P.N.; Shadduck, J.A.; Kang, C.Y.; Streilein, J.W.

    1982-08-01

    Homologous defective interfering (DI) particles protected adult Syrian hamsters against lethal systemic infection with vesicular stomatitis virus (VSV) serotype Indiana. The DI particles had to be biologically active, but did not have to be administered at the same inoculation site as the infectious virus. Serum and tissue levels of VSV postinoculation were significantly lower in DI-protected animals than in unprotected controls, suggesting that true autointerference was occurring. However, some aspects of protection also must be mediated through nonspecific mechanisms, since susceptible hamsters could be protected against VSV Indiana by coinjection with heterologous DI particles prepared from VSV serotype New Jersey or by simultaneous administration of polyinosinic acid-polycytidylic acid. By measuring serum levels of putative hamster interferon (type 1), we found that animals coinjected with VSV and DI particles or polyinosinic acid-polycytidylic acid produced significant levels of interferon. Since similarly high serum levels of interferon were measured in recipients of VSV alone (animals that eventually died from infection), there appeared to be no correlation between protection against lethal disease and induced levels of serum interferon. Instead, serum interferon levels correlated positively with amounts of VSV PFU found in serum and tissues of infected animals, the lowest levels being found in serum of animals protected with homologous DI particles. The data are consistent with the hypothesis that autointerference by DI particles as well as various host defense mechanisms (possibly including induction of interferon) participates in protecting hamsters against lethal VSV infection.

  12. H-2K/sup bm3/ mutation decreases the anti-vesicular stomatitis virus cytotoxic T-lymphocyte response

    Energy Technology Data Exchange (ETDEWEB)

    Bowman, M.R.; Lyles, D.; Parce, J.W.

    1986-03-01

    The authors have identified the substrain of mouse possessing the H-2K/sup bm3/ mutation as expressing a low anti-Vesicular Stomatitis Virus (VSV) cytotoxic T-lymphocyte (CTL) response upon secondary in vitro elicitation with ultraviolet light-inactivated virus. The CTL elicited from both the mutant, M505 (bm3), and the parental strain, C57B1/6(B6), kill B6 targets better than bm3 targets. Likewise, in cold target inhibition assays, unlabeled B6 cells inhibited better than unlabeled bm3 cells the recognition of /sup 51/Cr-labeled B6 targets regardless of whether B6 or bm3 CTL effector cells were used. CTL from both B6 and bm3 mice are H-2 restricted, virus specific, and possess the Thyl, Lyt2,3 phenotype. In addition, the effector cells use the H-2K molecule as the major restricting element. Both B6 and bm3 cells were shown by radiolabeling and immunoprecipitation to express an equivalent amount of G protein, the major viral surface glycoprotein, as well as equivalent amounts of H-2K. These data suggest the bm3 mutation in H-2K alters the association of H-2K with either the G protein or with a molecule on the CTL required for secondary in vitro elicitation.

  13. Oncolytic vesicular stomatitis virus in an immunocompetent model of MUC1-positive or MUC1-null pancreatic ductal adenocarcinoma.

    Science.gov (United States)

    Hastie, Eric; Besmer, Dahlia M; Shah, Nirav R; Murphy, Andrea M; Moerdyk-Schauwecker, Megan; Molestina, Carlos; Roy, Lopamudra Das; Curry, Jennifer M; Mukherjee, Pinku; Grdzelishvili, Valery Z

    2013-09-01

    Vesicular stomatitis virus (VSV) is a promising oncolytic agent against various malignancies. Here, for the first time, we tested VSV in vitro and in vivo in a clinically relevant, immunocompetent mouse model of pancreatic ductal adenocarcinoma (PDA). Our system allows the study of virotherapy against PDA in the context of overexpression (80% of PDA patients) or no expression of human mucin 1 (MUC1), a major marker for poor prognosis in patients. In vitro, we tested three VSV recombinants, wild-type VSV, VSV-green fluorescent protein (VSV-GFP), and a safe oncolytic VSV-ΔM51-GFP, against five mouse PDA cell lines that either expressed human MUC1 or were MUC1 null. All viruses demonstrated significant oncolytic abilities independent of MUC1 expression, although VSV-ΔM51-GFP was somewhat less effective in two PDA cell lines. In vivo administration of VSV-ΔM51-GFP resulted in significant reduction of tumor growth for tested mouse PDA xenografts (+MUC1 or MUC1 null), and antitumor efficacy was further improved when the virus was combined with the chemotherapeutic drug gemcitabine. The antitumor effect was transient in all tested groups. The developed system can be used to study therapies involving various oncolytic viruses and chemotherapeutics, with the goal of inducing tumor-specific immunity while preventing premature virus clearance.

  14. Attenuation of vesicular stomatitis virus infection of brain using antiviral drugs and an adeno-associated virus-interferon vector.

    Science.gov (United States)

    Wollmann, Guido; Paglino, Justin C; Maloney, Patrick R; Ahmadi, Sebastian A; van den Pol, Anthony N

    2015-01-15

    Vesicular stomatitis virus (VSV) shows promise as a vaccine-vector and oncolytic virus. However, reports of neurotoxicity of VSV remain a concern. We compared 12 antiviral compounds to control infection of VSV-CT9-M51 and VSV-rp30 using murine and human brain cultures, and in vivo mouse models. Inhibition of replication, cytotoxicity and infectivity was strongest with ribavirin and IFN-α and to some extent with mycophenolic acid, chloroquine, and adenine 9-β-d-arabinofuranoside. To generate continuous IFN exposure, we made an adeno-associated virus vector expressing murine IFN; AAV-mIFN-β protected mouse brain cells from VSV, as did a combination of IFN, ribavirin and chloroquine. Intracranial AAV-mIFN-β protected the brain against VSV-CT9-M51. In SCID mice bearing human glioblastoma, AAV-mIFN-β moderately enhanced survival. VSV-CT9-M51 doubled median survival when administered after AAV-mIFN-β; some surviving mice showed complete tumor destruction. Together, these data suggest that AAV-IFN or IFN with ribavirin and chloroquine provide an optimal anti-virus combination against VSV in the brain. Copyright © 2014 Elsevier Inc. All rights reserved.

  15. Digital sensing and sizing of vesicular stomatitis virus pseudotypes in complex media: a model for Ebola and Marburg detection.

    Science.gov (United States)

    Daaboul, George G; Lopez, Carlos A; Chinnala, Jyothsna; Goldberg, Bennett B; Connor, John H; Ünlü, M Selim

    2014-06-24

    Rapid, sensitive, and direct label-free capture and characterization of nanoparticles from complex media such as blood or serum will broadly impact medicine and the life sciences. We demonstrate identification of virus particles in complex samples for replication-competent wild-type vesicular stomatitis virus (VSV), defective VSV, and Ebola- and Marburg-pseudotyped VSV with high sensitivity and specificity. Size discrimination of the imaged nanoparticles (virions) allows differentiation between modified viruses having different genome lengths and facilitates a reduction in the counting of nonspecifically bound particles to achieve a limit-of-detection (LOD) of 5 × 10(3) pfu/mL for the Ebola and Marburg VSV pseudotypes. We demonstrate the simultaneous detection of multiple viruses in a single sample (composed of serum or whole blood) for screening applications and uncompromised detection capabilities in samples contaminated with high levels of bacteria. By employing affinity-based capture, size discrimination, and a "digital" detection scheme to count single virus particles, we show that a robust and sensitive virus/nanoparticle sensing assay can be established for targets in complex samples. The nanoparticle microscopy system is termed the Single Particle Interferometric Reflectance Imaging Sensor (SP-IRIS) and is capable of high-throughput and rapid sizing of large numbers of biological nanoparticles on an antibody microarray for research and diagnostic applications.

  16. Dynamics of melanoma tumor therapy with vesicular stomatitis virus: explaining the variability in outcomes using mathematical modeling.

    Science.gov (United States)

    Rommelfanger, D M; Offord, C P; Dev, J; Bajzer, Z; Vile, R G; Dingli, D

    2012-05-01

    Tumor selective, replication competent viruses are being tested for cancer gene therapy. This approach introduces a new therapeutic paradigm due to potential replication of the therapeutic agent and induction of a tumor-specific immune response. However, the experimental outcomes are quite variable, even when studies utilize highly inbred strains of mice and the same cell line and virus. Recognizing that virotherapy is an exercise in population dynamics, we utilize mathematical modeling to understand the variable outcomes observed when B16ova malignant melanoma tumors are treated with vesicular stomatitis virus in syngeneic, fully immunocompetent mice. We show how variability in the initial tumor size and the actual amount of virus delivered to the tumor have critical roles on the outcome of therapy. Virotherapy works best when tumors are small, and a robust innate immune response can lead to superior tumor control. Strategies that reduce tumor burden without suppressing the immune response and methods that maximize the amount of virus delivered to the tumor should optimize tumor control in this model system.

  17. Oncolytic Recombinant Vesicular Stomatitis Virus (VSV) Is Nonpathogenic and Nontransmissible in Pigs, a Natural Host of VSV.

    Science.gov (United States)

    Velazquez-Salinas, Lauro; Naik, Shruthi; Pauszek, Steven J; Peng, Kah-Whye; Russell, Stephen J; Rodriguez, Luis L

    2017-06-01

    Vesicular stomatitis virus (VSV) is a negative-stranded RNA virus that naturally causes disease in livestock including horses, cattle and pigs. The two main identified VSV serotypes are New Jersey (VSNJV) and Indiana (VSIV). VSV is a rapidly replicating, potently immunogenic virus that has been engineered to develop novel oncolytic therapies for cancer treatment. Swine are a natural host for VSV and provide a relevant and well-established model, amenable to biological sampling to monitor virus shedding and neutralizing antibodies. Previous reports have documented the pathogenicity and transmissibility of wild-type isolates and recombinant strains of VSIV and VSNJV using the swine model. Oncolytic VSV engineered to express interferon-beta (IFNβ) and the sodium iodide symporter (NIS), VSV-IFNβ-NIS, has been shown to be a potent new therapeutic agent inducing rapid and durable tumor remission following systemic therapy in preclinical mouse models. VSV-IFNβ-NIS is currently undergoing clinical evaluation for the treatment of advanced cancer in human and canine patients. To support clinical studies and comprehensively assess the risk of transmission to susceptible species, we tested the pathogenicity and transmissibility of oncolytic VSV-IFNβ-NIS using the swine model. Following previously established protocols to evaluate VSV pathogenicity, intradermal inoculation with 107 TCID50 VSV-IFNβ-NIS caused no observable symptoms in pigs. There was no detectable shedding of infectious virus in VSV-IFNβ-NIS in biological excreta of inoculated pigs or exposed naive pigs kept in direct contact throughout the experiment. VSV-IFNβ-NIS inoculated pigs became seropositive for VSV antibodies, while contact pigs displayed no symptoms of VSV infection, and importantly did not seroconvert. These data indicate that oncolytic VSV is both nonpathogenic and not transmissible in pigs, a natural host. These findings support further clinical development of oncolytic VSV-IFNβ-NIS as a

  18. The length of vesicular stomatitis virus particles dictates a need for actin assembly during clathrin-dependent endocytosis.

    Directory of Open Access Journals (Sweden)

    David K Cureton

    2010-09-01

    Full Text Available Microbial pathogens exploit the clathrin endocytic machinery to enter host cells. Vesicular stomatitis virus (VSV, an enveloped virus with bullet-shaped virions that measure 70 x 200 nm, enters cells by clathrin-dependent endocytosis. We showed previously that VSV particles exceed the capacity of typical clathrin-coated vesicles and instead enter through endocytic carriers that acquire a partial clathrin coat and require local actin filament assembly to complete vesicle budding and internalization. To understand why the actin system is required for VSV uptake, we compared the internalization mechanisms of VSV and its shorter (75 nm long defective interfering particle, DI-T. By imaging the uptake of individual particles into live cells, we found that, as with parental virions, DI-T enters via the clathrin endocytic pathway. Unlike VSV, DI-T internalization occurs through complete clathrin-coated vesicles and does not require actin polymerization. Since VSV and DI-T particles display similar surface densities of the same attachment glycoprotein, we conclude that the physical properties of the particle dictate whether a virus-containing clathrin pit engages the actin system. We suggest that the elongated shape of a VSV particle prevents full enclosure by the clathrin coat and that stalling of coat assembly triggers recruitment of the actin machinery to finish the internalization process. Since some enveloped viruses have pleomorphic particle shapes and sizes, our work suggests that they may use altered modes of endocytic uptake. More generally, our findings show the importance of cargo geometry for specifying cellular entry modes, even when the receptor recognition properties of a ligand are maintained.

  19. Effects of intravenously administered recombinant vesicular stomatitis virus (VSV(deltaM51)) on multifocal and invasive gliomas.

    Science.gov (United States)

    Lun, XueQing; Senger, Donna L; Alain, Tommy; Oprea, Andra; Parato, Kelley; Stojdl, Dave; Lichty, Brian; Power, Anthony; Johnston, Randal N; Hamilton, Mark; Parney, Ian; Bell, John C; Forsyth, Peter A

    2006-11-01

    An ideal virus for the treatment of cancer should have effective delivery into multiple sites within the tumor, evade immune responses, produce rapid viral replication, spread within the tumor, and infect multiple tumors. Vesicular stomatitis virus (VSV) has been shown to be an effective oncolytic virus in a variety of tumor models, and mutations in the matrix (M) protein enhance VSV's effectiveness in animal models. We evaluated the susceptibility of 14 glioma cell lines to infection and killing by mutant strain VSV(deltaM51), which contains a single-amino acid deletion in the M protein. We also examined the activity and safety of this strain against the U87 and U118 experimental models of human malignant glioma in nude mice and analyzed the distribution of the virus in the brains of U87 tumor-bearing mice using fluorescence labeling. Finally, we examined the effect of VSV(deltaM51) on 15 primary human gliomas cultured from surgical specimens. All statistical tests were two-sided. All 14 glioma cell lines were susceptible to VSV(deltaM51) infection and killing. Intratumoral administration of VSV(deltaM51) produced marked regression of malignant gliomas in nude mice. When administered systemically, live VSV(deltaM51) virus, as compared with dead virus, statistically significantly prolonged survival of mice with unilateral U87 tumors (median survival: 113 versus 46 days, P = .0001) and bilateral U87 tumors (median survival: 73 versus 46 days, P = .0025). VSV(deltaM51) infected multifocal gliomas, invasive glioma cells that migrated beyond the main glioma, and all 15 primary human gliomas. There was no evidence of toxicity. Systemically delivered VSV(deltaM51) was an effective and safe oncolytic agent against laboratory models of multifocal and invasive malignant gliomas, the most challenging clinical manifestations of this disease.

  20. Heterogeneous Nuclear Ribonucleoprotein K Supports Vesicular Stomatitis Virus Replication by Regulating Cell Survival and Cellular Gene Expression

    Science.gov (United States)

    Dinh, Phat X.; Das, Anshuman; Franco, Rodrigo

    2013-01-01

    The heterogeneous nuclear ribonucleoprotein K (hnRNP K) is a member of the family of hnRNPs and was recently shown in a genome-wide small interfering RNA (siRNA) screen to support vesicular stomatitis virus (VSV) growth. To decipher the role of hnRNP K in VSV infection, we conducted studies which suggest that the protein is required for VSV spreading. Virus binding to cells, entry, and nucleocapsid uncoating steps were not adversely affected in the absence of hnRNP K, whereas viral genome transcription and replication were reduced slightly. These results indicate that hnRNP K is likely involved in virus assembly and/or release from infected cells. Further studies showed that hnRNP K suppresses apoptosis of virus-infected cells, resulting in increased cell survival during VSV infection. The increased survival of the infected cells was found to be due to the suppression of proapoptotic proteins such as Bcl-XS and Bik in a cell-type-dependent manner. Additionally, depletion of hnRNP K resulted in not only significantly increased levels of T-cell-restricted intracellular antigen 1 (TIA1) but also switching of the expression of the two isoforms of the protein (TIA1a and TIA1b), both of which inhibited VSV replication. hnRNP K was also found to support expression of several cellular proteins known to be required for VSV infection. Overall, our studies demonstrate hnRNP K to be a multifunctional protein that supports VSV infection via its role(s) in suppressing apoptosis of infected cells, inhibiting the expression of antiviral proteins, and maintaining the expression of proteins required for the virus. PMID:23843646

  1. The Immune Response to a Vesicular Stomatitis Virus Vaccine Vector Is Independent of Particulate Antigen Secretion and Protein Turnover Rate

    Science.gov (United States)

    Cobleigh, Melissa A.; Bradfield, Clinton; Liu, Yuanjie; Mehta, Anand

    2012-01-01

    Vesicular stomatitis virus (VSV) is a highly cytopathic virus being developed as a vaccine vector due to its ability to induce strong protective T cell and antibody responses after a single dose. However, little is known regarding the mechanisms underlying the potent immune responses elicited by VSV. We previously generated a VSV vector expressing the hepatitis B virus middle envelope surface glycoprotein (MS) that induces strong MS-specific T cell and antibody responses in mice. After synthesis in the cytoplasm, the MS protein translocates to the endoplasmic reticulum, where it forms subviral particles that are secreted from the cell. To better understand the contributions of secreted and intracellular protein to the VSV-induced immune response, we produced a vector expressing a secretion-deficient MS mutant (MSC69A) and compared the immunogenicity of this vector to that of the wild-type VSV-MS vector in mice. As expected, the MSC69A protein was not secreted from VSV-infected cells and displayed enhanced proteasome-mediated degradation. Surprisingly, despite these differences in intracellular protein processing, the T cell and antibody responses generated to MSC69A were comparable to those elicited by virus expressing wild-type MS protein. Therefore, when it is expressed from VSV, the immune responses to MS are independent of particulate antigen secretion and the turnover rate of cytoplasmic protein. These results are consistent with a model in which the immune responses to VSV are strongly influenced by the replication cycle of the vector and demonstrate that characteristics of the vector have the capacity to affect vaccine efficacy more than do the properties of the antigen itself. PMID:22345454

  2. The Length of Vesicular Stomatitis Virus Particles Dictates a Need for Actin Assembly during Clathrin-Dependent Endocytosis

    Science.gov (United States)

    Cureton, David K.; Massol, Ramiro H.; Whelan, Sean P. J.; Kirchhausen, Tomas

    2010-01-01

    Microbial pathogens exploit the clathrin endocytic machinery to enter host cells. Vesicular stomatitis virus (VSV), an enveloped virus with bullet-shaped virions that measure 70×200 nm, enters cells by clathrin-dependent endocytosis. We showed previously that VSV particles exceed the capacity of typical clathrin-coated vesicles and instead enter through endocytic carriers that acquire a partial clathrin coat and require local actin filament assembly to complete vesicle budding and internalization. To understand why the actin system is required for VSV uptake, we compared the internalization mechanisms of VSV and its shorter (75 nm long) defective interfering particle, DI-T. By imaging the uptake of individual particles into live cells, we found that, as with parental virions, DI-T enters via the clathrin endocytic pathway. Unlike VSV, DI-T internalization occurs through complete clathrin-coated vesicles and does not require actin polymerization. Since VSV and DI-T particles display similar surface densities of the same attachment glycoprotein, we conclude that the physical properties of the particle dictate whether a virus-containing clathrin pit engages the actin system. We suggest that the elongated shape of a VSV particle prevents full enclosure by the clathrin coat and that stalling of coat assembly triggers recruitment of the actin machinery to finish the internalization process. Since some enveloped viruses have pleomorphic particle shapes and sizes, our work suggests that they may use altered modes of endocytic uptake. More generally, our findings show the importance of cargo geometry for specifying cellular entry modes, even when the receptor recognition properties of a ligand are maintained. PMID:20941355

  3. Interferon induction by viruses. XV. Biological characteristics of interferon induction-suppressing particles of vesicular stomatitis virus

    Energy Technology Data Exchange (ETDEWEB)

    Marcus, P.I.; Sekellick, M.J.

    1987-06-01

    A single interferon (IFN) induction-suppressing particle (ISP) of vesicular stomatitis virus (VSV) blocked completely the yield of IFN in a cell otherwise programmed to produce IFN. With mouse L cells as hosts, one lethal hit of UV radiation (D37 = 52.5 ergs/mm2) to the VSV genome sufficed to inactivate ISP activity; however, with ''aged'' primary chick embryo cells as hosts, it took 198 lethal hits (D37 = 10,395 ergs/mm2). ISP expression in chick cells did not require virus replication or amplified RNA synthesis, but did involve functional virion-associated L protein. ISP in chick cells also were capable of inhibiting, in a multiplicity-dependent manner, the plaquing efficiency of two viruses that require cellular polymerase II (pol II) for replication, e.g., pseudorabies and influenza. The refractory state to IFN inducibility that resulted from infection of chick cells with ISP (VSV tsO5 (UV = 100 hits)) was still extant after 6 days. In contrast, the plaquing efficiency of pseudorabies virus returned to control levels by 5 h after ISP infection. Chick cells infected with UV ISP remained viable, served as hosts for the replication of other viruses, and could be subcultured. Models are presented to account for these contrasting effects. The involvement of viral plus-strand leader RNA as an inhibitor of cellular pol II-dependent RNA synthesis, and the multifunctional activities of the virion-associated L protein, are discussed as possible molecules involved in the action of ISP in chick cells.

  4. Vesicular Stomatitis Virus Has Extensive Oncolytic Activity against Human Sarcomas: Rare Resistance Is Overcome by Blocking Interferon Pathways ▿

    Science.gov (United States)

    Paglino, Justin C.; van den Pol, Anthony N.

    2011-01-01

    Oncolytic viruses have been tested against many carcinomas of ectodermal and endodermal origin; however, sarcomas, arising from mesoderm, have received relatively little attention. Using 13 human sarcomas representing seven tumor types, we assessed the efficiency of infection, cytolysis, and replication of green fluorescent protein (GFP)-expressing vesicular stomatitis virus (VSV) and its oncolytically enhanced mutant VSV-rp30a. Both viruses efficiently infected and killed 12 of 13 sarcomas. VSV-rp30a showed a faster rate of infection and replication. In vitro and in vivo, VSV was selective for sarcomas compared with normal mesoderm. A single intravenous injection of VSV-rp30a selectively infected all subcutaneous human sarcomas tested in mice and arrested the growth of tumors that otherwise grew 11-fold. In contrast to other sarcomas, synovial sarcoma SW982 demonstrated remarkable resistance, even to high titers of virus (multiplicity of infection [MOI] of 100). We found no dysfunction in VSV binding or internalization. SW982 also resisted infection by human cytomegalovirus and Sindbis virus, suggesting a virus resistance mechanism based on an altered antiviral state. Quantitative reverse transcriptase (qRT)-PCR analysis revealed a heightened basal expression of interferon-stimulated genes (ISGs). Pretreatment, but not cotreatment, with interferon attenuators valproate, Jak1 inhibitor, or vaccinia virus B18R protein rendered SW982 highly susceptible, and this correlated with downregulation of ISG expression. Jak1 inhibitor pretreatment also enhanced susceptibility in moderately VSV-resistant liposarcoma and bladder carcinoma. Overall, we find that the potential efficacy of VSV as an oncolytic agent extends to nonhematologic mesodermal tumors and that unusually strong resistance to VSV oncolysis can be overcome with interferon attenuators. PMID:21734048

  5. A freeze frame view of vesicular stomatitis virus transcription defines a minimal length of RNA for 5' processing.

    Science.gov (United States)

    Tekes, Gergely; Rahmeh, Amal A; Whelan, Sean P J

    2011-06-01

    The RNA synthesis machinery of vesicular stomatitis virus (VSV) comprises the genomic RNA encapsidated by the viral nucleocapsid protein (N) and associated with the RNA dependent RNA polymerase, the viral components of which are a large protein (L) and an accessory phosphoprotein (P). The 241 kDa L protein contains all the enzymatic activities necessary for synthesis of the viral mRNAs, including capping, cap methylation and polyadenylation. Those RNA processing reactions are intimately coordinated with nucleotide polymerization such that failure to cap results in termination of transcription and failure to methylate can result in hyper polyadenylation. The mRNA processing reactions thus serve as a critical check point in viral RNA synthesis which may control the synthesis of incorrectly modified RNAs. Here, we report the length at which viral transcripts first gain access to the capping machinery during synthesis. By reconstitution of transcription in vitro with highly purified recombinant polymerase and engineered templates in which we omitted sites for incorporation of UTP, we found that transcripts that were 30-nucleotides in length were uncapped, whereas those that were 31-nucleotides in length contained a cap structure. The minimal RNA length required for mRNA cap addition was also sufficient for methylation since the 31-nucleotide long transcripts were methylated at both ribose-2'-O and guanine-N-7 positions. This work provides insights into the spatial relationship between the active sites for the RNA dependent RNA polymerase and polyribonucleotidyltransferase responsible for capping of the viral RNA. We combine the present findings with our recently described electron microscopic structure of the VSV polymerase and propose a model of how the spatial arrangement of the capping activities of L may influence nucleotide polymerization.

  6. Efficacy of Vesicular Stomatitis Virus-Ebola Virus Postexposure Treatment in Rhesus Macaques Infected With Ebola Virus Makona.

    Science.gov (United States)

    Marzi, Andrea; Hanley, Patrick W; Haddock, Elaine; Martellaro, Cynthia; Kobinger, Gary; Feldmann, Heinz

    2016-10-15

    The Ebola virus (EBOV) epidemic in West Africa increased the focus on vaccine development against this hemorrhagic fever-causing pathogen, and as a consequence human clinical trials for a few selected platforms were accelerated. One of these vaccines is vesicular stomatitis virus (VSV)-EBOV, also known as rVSV-ZEBOV, a fast-acting vaccine against EBOV and so far the only vaccine with reported efficacy against EBOV infections in humans in phase III clinical trials. In this study, we analyzed the potential of VSV-EBOV for postexposure treatment of rhesus macaques infected with EBOV-Makona. We treated groups of animals with 1 dose of VSV-EBOV either in a single injection at 1 or 24 hours after EBOV exposure or with 2 injections, half the dose at each time point; 1 control group received the same dose of the VSV-based Marburg virus vaccine at both time points; another group remained untreated. Although all untreated animals succumbed to EBOV infection, 33%-67% of the animals in each treatment group survived the infection, including the group treated with the VSV-based Marburg virus vaccine. This result suggests that protection from postexposure vaccination may be antigen unspecific and due rather to an early activation of the innate immune system. In conclusion, VSV-EBOV remains a potent and fast-acting prophylactic vaccine but demonstrates only limited efficacy in postexposure treatment. Published by Oxford University Press for the Infectious Diseases Society of America 2016. This work is written by (a) US Government employee(s) and is in the public domain in the US.

  7. Inactivation of a Human Norovirus Surrogate, Human Norovirus Virus-Like Particles, and Vesicular Stomatitis Virus by Gamma Irradiation ▿

    Science.gov (United States)

    Feng, Kurtis; Divers, Erin; Ma, Yuanmei; Li, Jianrong

    2011-01-01

    Gamma irradiation is a nonthermal processing technology that has been used for the preservation of a variety of food products. This technology has been shown to effectively inactivate bacterial pathogens. Currently, the FDA has approved doses of up to 4.0 kGy to control food-borne pathogens in fresh iceberg lettuce and spinach. However, whether this dose range effectively inactivates food-borne viruses is less understood. We have performed a systematic study on the inactivation of a human norovirus surrogate (murine norovirus 1 [MNV-1]), human norovirus virus-like particles (VLPs), and vesicular stomatitis virus (VSV) by gamma irradiation. We demonstrated that MNV-1 and human norovirus VLPs were resistant to gamma irradiation. For MNV-1, only a 1.7- to 2.4-log virus reduction in fresh produce at the dose of 5.6 kGy was observed. However, VSV was more susceptible to gamma irradiation, and a 3.3-log virus reduction at a dose of 5.6 kGy in Dulbecco's modified Eagle medium (DMEM) was achieved. We further demonstrated that gamma irradiation disrupted virion structure and degraded viral proteins and genomic RNA, which resulted in virus inactivation. Using human norovirus VLPs as a model, we provide the first evidence that the capsid of human norovirus has stability similar to that of MNV-1 after exposure to gamma irradiation. Overall, our results suggest that viruses are much more resistant to irradiation than bacterial pathogens. Although gamma irradiation used to eliminate the virus contaminants in fresh produce by the FDA-approved irradiation dose limits seems impractical, this technology may be practical to inactivate viruses for other purposes, such as sterilization of medical equipment. PMID:21441330

  8. Vesicular stomatitis virus expressing interferon-? is oncolytic and promotes antitumor immune responses in a syngeneic murine model of non-small cell lung cancer

    OpenAIRE

    Patel, Manish R.; Jacobson, Blake A.; Ji, Yan; Drees, Jeremy; Tang, Shaogeng; Xiong, Kerry; Wang, Hengbing; Prigge, Jennifer E.; Dash, Alexander S.; Kratzke, Andrea K.; Mesev, Emily; Etchison, Ryan; Federspiel, Mark J.; Russell, Stephen J; Kratzke, Robert A

    2015-01-01

    Vesicular stomatitis virus (VSV) is a potent oncolytic virus for many tumors. VSV that produces interferon-? (VSV-IFN?) is now in early clinical testing for solid tumors. Here, the preclinical activity of VSV and VSV-IFN? against non-small cell lung cancer (NSCLC) is reported. NSCLC cell lines were treated in vitro with VSV expressing green fluorescence protein (VSV-GFP) and VSV-IFN?. VSV-GFP and VSV-IFN? were active against NSCLC cells. JAK/STAT inhibition with ruxolitinib re-sensitized resi...

  9. Vesicular Stomatitis Virus Pseudotyped with Ebola Virus Glycoprotein Serves as a Highly Protective, Non-infectious Vaccine Against Ebola Virus Challenge

    Science.gov (United States)

    2016-07-01

    parainfluenza virus type 3, 62 rabies virus and vesicular stomatitis virus (VSV) (reviewed in (7, 11)). Surprisingly, EBOV GP 63 pseudovirions have not been...Modified Eagle Medium (DMEM) with 10% fetal bovine serum (FBS) and 1% 111 penicillin/streptomycin. The pcDNA3.1 expression plasmids for EBOV GP...198 bovine serum albumin (BSA), incubated with serial dilutions (1:1000, 1:10,000 and 1:1000,000) 199 of serum for overnight at 4°C, probed with 1

  10. Acute reactogenicity after intramuscular immunization with recombinant vesicular stomatitis virus is linked to production of IL-1β.

    Directory of Open Access Journals (Sweden)

    Kathleen Athearn

    Full Text Available Vaccines based on live viruses are attractive because they are immunogenic, cost-effective, and can be delivered by multiple routes. However, live virus vaccines also cause reactogenic side effects such as fever, myalgia, and injection site pain that have reduced their acceptance in the clinic. Several recent studies have linked vaccine-induced reactogenic side effects to production of the pro-inflammatory cytokine interleukin-1β (IL-1β in humans. Our objective was therefore to determine whether IL-1β contributed to pathology after immunization with recombinant vesicular stomatitis virus (rVSV vaccine vectors, and if so, to identify strategies by which IL-1β mediated pathology might be reduced without compromising immunogenicity. We found that an rVSV vaccine induced local and systemic production of IL-1β in vivo, and that accumulation of IL-1β correlated with acute pathology after rVSV immunization. rVSV-induced pathology was reduced in mice deficient in the IL-1 receptor Type I, but the IL-1R-/- mice were fully protected from lethal rechallenge with a high dose of VSV. This result demonstrated that IL-1 contributed to reactogenicity of the rVSV, but was dispensable for induction of protective immunity. The amount of IL-1β detected in mice deficient in either caspase-1 or the inflammasome adaptor molecule ASC after rVSV immunization was not significantly different than that produced by wild type animals, and caspase-1-/- and ASC-/- mice were only partially protected from rVSV-induced pathology. Those data support the idea that some of the IL-1β expressed in vivo in response to VSV may be activated by a caspase-1 and ASC-independent mechanism. Together these results suggest that rVSV vectors engineered to suppress the induction of IL-1β, or signaling through the IL-1R would be less reactogenic in vivo, but would retain their immunogenicity and protective capacity. Such rVSV would be highly desirable as either vaccine vectors or

  11. 5'-Phospho-RNA Acceptor Specificity of GDP Polyribonucleotidyltransferase of Vesicular Stomatitis Virus in mRNA Capping.

    Science.gov (United States)

    Ogino, Minako; Ogino, Tomoaki

    2017-03-15

    The GDP polyribonucleotidyltransferase (PRNTase) domain of the multifunctional L protein of rhabdoviruses, such as vesicular stomatitis virus (VSV) and rabies virus, catalyzes the transfer of 5'-phospho-RNA (pRNA) from 5'-triphospho-RNA (pppRNA) to GDP via a covalent enzyme-pRNA intermediate to generate a 5'-cap structure (GpppA). Here, using an improved oligo-RNA capping assay with the VSV L protein, we showed that the Michaelis constants for GDP and pppAACAG (VSV mRNA-start sequence) are 0.03 and 0.4 μM, respectively. A competition assay between GDP and GDP analogues in the GpppA formation and pRNA transfer assay using GDP analogues as pRNA acceptors indicated that the PRNTase domain recognizes the C-2-amino group, but not the C-6-oxo group, N-1-hydrogen, or N-7-nitrogen, of GDP for the cap formation. 2,6-Diaminopurine-riboside (DAP), 7-deazaguanosine (7-deaza-G), and 7-methylguanosine (m7G) diphosphates efficiently accepted pRNA, resulting in the formation of DAPpppA, 7-deaza-GpppA, and m7GpppA (cap 0), respectively. Furthermore, either the 2'- or 3'-hydroxyl group of GDP was found to be required for efficient pRNA transfer. A 5'-diphosphate form of antiviral ribavirin weakly inhibited the GpppA formation but did not act as a pRNA acceptor. These results indicate that the PRNTase domain has a unique guanosine-binding mode different from that of eukaryotic mRNA capping enzyme, guanylyltransferase. IMPORTANCE mRNAs of nonsegmented negative-strand (NNS) RNA viruses, such as VSV, possess a fully methylated cap structure, which is required for mRNA stability, efficient translation, and evasion of antiviral innate immunity in host cells. GDP polyribonucleotidyltransferase (PRNTase) is an unconventional mRNA capping enzyme of NNS RNA viruses that is distinct from the eukaryotic mRNA capping enzyme, guanylyltransferase. In this study, we studied the pRNA acceptor specificity of VSV PRNTase using various GDP analogues and identified chemical groups of GDP as essential

  12. Intrinsic innate immunity fails to control herpes simplex virus and vesicular stomatitis virus replication in sensory neurons and fibroblasts.

    Science.gov (United States)

    Rosato, Pamela C; Leib, David A

    2014-09-01

    Herpes simplex virus 1 (HSV-1) establishes lifelong latent infections in the sensory neurons of the trigeminal ganglia (TG), wherein it retains the capacity to reactivate. The interferon (IFN)-driven antiviral response is critical for the control of HSV-1 acute replication. We therefore sought to further investigate this response in TG neurons cultured from adult mice deficient in a variety of IFN signaling components. Parallel experiments were also performed in fibroblasts isolated concurrently. We showed that HSV-1 replication was comparable in wild-type (WT) and IFN signaling-deficient neurons and fibroblasts. Unexpectedly, a similar pattern was observed for the IFN-sensitive vesicular stomatitis virus (VSV). Despite these findings, TG neurons responded to IFN-β pretreatment with STAT1 nuclear localization and restricted replication of both VSV and an HSV-1 strain deficient in γ34.5, while wild-type HSV-1 replication was unaffected. This was in contrast to fibroblasts in which all viruses were restricted by the addition of IFN-β. Taken together, these data show that adult TG neurons can mount an effective antiviral response only if provided with an exogenous source of IFN-β, and HSV-1 combats this response through γ34.5. These results further our understanding of the antiviral response of neurons and highlight the importance of paracrine IFN-β signaling in establishing an antiviral state. Herpes simplex virus 1 (HSV-1) is a ubiquitous virus that establishes a lifelong latent infection in neurons. Reactivation from latency can cause cold sores, blindness, and death from encephalitis. Humans with deficiencies in innate immunity have significant problems controlling HSV infections. In this study, we therefore sought to elucidate the role of neuronal innate immunity in the control of viral infection. Using neurons isolated from mice, we found that the intrinsic capacity of neurons to restrict virus replication was unaffected by the presence or absence of

  13. Strains of Lentinula edodes suppress growth of phytopathogenic fungi and inhibit Alagoas serotype of vesicular stomatitis virus Linhagens de Lentinula edodes inibem fungos fitopatogênicos e o vírus da estomatite vesicular, sorotipo Alagoas

    Directory of Open Access Journals (Sweden)

    Selma H. Sasaki

    2001-03-01

    Full Text Available Four Lentinula edodes strains (Le10, 46, K2, Assai were assessed for their antagonistic effect on four filamentous fungus species of agricultural importance (Helminthosporium euphorbiae, Helminthosporium sp, Fusarium solani and Phomopsis sojae and on Alagoas serotype of Vesicular Stomatitis Virus (VSA. The L. edodes strains studied had variable effects on the filamentous fungi and on VSA. The K2 and Le10 strains were antagonistic on the fungi assessed and the 46 and K2 strains were efficient on the Vesicular Stomatitis Virus. The results widened the list of beneficial effects of L. edodes on the control and prevention of animal pathogenic virus and filamentous fungi.Quatro linhagens de Lentinula edodes (Le10, 46, K2, ASSAI foram avaliadas quanto ao seu efeito inibitório sobre quatro espécies de fungos filamentosos de importância agrícola (Helminthosporium euphorbiae, Helminthosporium sp., Fusarium solani, Phomopsis sojae e sobre o sorotipo Alagoas vírus da estomatite vesicular (VSA. Foi observado que as linhagens de L. edodes estudadas apresentaram variabilidade quanto ao seu efeito, tanto sobre os fungos filamentosos quanto sobre o vírus VSA. As linhagens K2 e Le10 apresentaram-se antagônicas sobre os fungos e as linhagens 46 e K2 foram eficientes na inibição do vírus VSA. Os resultados obtidos permitem ampliar a lista de efeitos benéficos de algumas linhagens de L. edodes no controle e prevenção de vírus patogênicos animais e de fungos filamentosos.

  14. Development of monoclonal antibody-linked ELISA for sero-diagnosis of vesicular stomatitis virus (VSV-IN) using baculovirus expressed glycoprotein.

    Science.gov (United States)

    Kweon, Chang Hee; Kwon, Byung Joon; Kim, In Joong; Lee, Se Young; Ko, Young Joon

    2005-12-01

    The gene encoding the envelope glycoprotein (GP) of vesicular stomatitis virus serotype, Indiana (VSV-IN), was expressed under the polyhedron promoter of baculovirus. The recombinant GP was applied as a diagnostic antigen for the detection of cattle and horse antibodies to VSV. In addition, the neutralizing monoclonal antibody (Mab) to GP of VSV-IN was used as trapping antibody in a Mab-linked indirect ELISA (MLI-ELISA) or detecting antibody in a Mab-linked competitive ELISA (MLC-ELISA). The diagnostic efficiencies of MLI-ELISA and MLC-ELISA were evaluated with currently available C-ELISA from OIE reference laboratory for vesicular stomatitis as a gold standard by using VSV-positive equine sera and negative bovine sera vaccinated against foot-and-mouth disease (FMD) in the field. When naturally infected equine sera and FMDV vaccinated bovine sera were tested, MLI-ELISA and MLC-ELISA showed relative sensitivities of 80% and 95% with relative specificity of 97% and 99%, respectively. However, both ELISAs cross-reacted with equine sera against New Jersey (VSV-NJ) serotype. The comparison of the two ELISAs revealed that MLC-ELISA was relatively more sensitive and specific than MLI-ELISA, indicating that MLC-ELISA can be applied to sero-diagnosis for VSV-IN infection.

  15. Experimental transmission of vesicular stomatitis New Jersey virus from Simulium vittatum to cattle: clinical outcome is influenced by site of insect feeding.

    Science.gov (United States)

    Mead, D G; Lovett, K Rainwater; Murphy, M D; Pauszek, S J; Smoliga, G; Gray, E W; Noblet, R; Overmyer, J; Rodriguez, L L

    2009-07-01

    Vesicular stomatitis New Jersey virus (VSNJV) is an insect-transmitted Rhabdovirus causing vesicular disease in domestic livestock including cattle, horses, and pigs. Natural transmission during epidemics remains poorly understood, particularly in cattle, one of the most affected species during outbreaks. This study reports the first successful transmission of VSNJV to cattle by insect bite resulting in clinical disease. When infected black flies (Simulium vittatum Zetterstedt) fed at sites where VS lesions are usually observed (mouth, nostrils, and foot coronary band), infection occurred, characterized by local viral replication, vesicular lesions, and high neutralizing antibody titers (> 1: 256). Viral RNA was detected up to 9 d postinfection in tissues collected during necropsy from lesion sites and lymph nodes draining those sites. Interestingly, when flies were allowed to feed on flank or neck skin, viral replication was poor, lesions were not observed, and low levels of neutralizing antibodies (range, 1:8-1:32) developed. Viremia was never observed in any of the animals and infectious virus was not recovered from tissues on necropsies performed between 8 and 27 d postinfection. Demonstration that VSNJV transmission to cattle by infected black flies can result in clinical disease contributes to a better understanding of the epidemiology and potential prevention and control methods for this important disease.

  16. Identification of a new region in the vesicular stomatitis virus L polymerase protein which is essential for mRNA cap methylation.

    Science.gov (United States)

    Grdzelishvili, Valery Z; Smallwood, Sherin; Tower, Dallas; Hall, Richard L; Hunt, D Margaret; Moyer, Sue A

    2006-07-05

    The vesicular stomatitis virus (VSV) L polymerase protein possesses two methyltransferase (MTase) activities, which catalyze the methylation of viral mRNA cap structures at the guanine-N7 and 2'-O-adenosine positions. To identify L sequences required for the MTase activities, we analyzed a host range (hr) and temperature-sensitive (ts) mutant of VSV, hr8, which was defective in mRNA cap methylation. Sequencing hr8 identified five amino acid substitutions, all residing in the L protein. Recombinant VSV were generated with each of the identified L mutations, and the presence of a single G1481R substitution in L, located between conserved domains V and VI, was sufficient to produce a dramatic reduction (about 90%) in overall mRNA methylation. Cap analysis showed residual guanine-N7 methylation and reduced 2'-O-adenosine methylation, identical to that of the original hr8 virus. When recombinant viruses were tested for virus growth under conditions that were permissive and nonpermissive for the hr8 mutant, the same single L mutation, G1481R, was solely responsible for both the hr and ts phenotypes. A spontaneous suppressor mutant of the rG1481R virus that restored both growth on nonpermissive cells and cap methylation was identified and mapped to a single change, L1450I, in L. Site-directed mutagenesis of the region between domains V and VI, amino acids 1419-1672 of L, followed by the rescue of recombinant viruses identified five additional virus mutants, K1468A, R1478A/D1479A, G1481A, G1481N, and G1672A, that were all hr and defective in mRNA cap methylation. Thus, in addition to the previously characterized domain VI [Grdzelishvili, V.Z., Smallwood, S., Tower, D., Hall, R.L., Hunt, D.M., Moyer, S.A., 2005. A single amino acid change in the L-polymerase protein of vesicular stomatitis virus completely abolishes viral mRNA cap methylation. J. Virol. 79, 7327-7337; Li, J., Fontaine-Rodriguez, E.C., Whelan, S.P., 2005. Amino acid residues within conserved domain VI of the

  17. Alteration of vesicular stomatitis virus L and NS proteins by uv irradiation: implications for the mechanism of host cell shut-off

    Energy Technology Data Exchange (ETDEWEB)

    Whitaker-Dowling, P.; Youngner, J.S.

    1988-05-01

    When purified, (/sup 35/S)methionine-labeled vesicular stomatitis virus (VSV) was exposed to ultraviolet light, an irradiation-induced change in the viral proteins was detected by SDS-polyacrylamide gel electrophoresis and immunoblotting. With dose of uv irradiation in the same range as that required to inactivate VSV leader RNA, a loss occurred in the bands corresponding to the L and NS proteins concomitant with the appearance of several new bands of radioactivity throughout the gel. This alteration of viral proteins correlated with the loss of ability of the virus to inhibit host macromolecular synthesis. In light of these results, the role that has been ascribed to the VSV leader RNA in VSV-mediated host shut-off needs to be reevaluated.

  18. Phylogeographic characteristics of vesicular stomatitis New Jersey viruses circulating in Mexico from 2005 to 2011 and their relationship to epidemics in the United States.

    Science.gov (United States)

    Velazquez-Salinas, Lauro; Pauszek, Steven J; Zarate, Selene; Basurto-Alcantara, Francisco J; Verdugo-Rodriguez, Antonio; Perez, Andres M; Rodriguez, Luis L

    2014-01-20

    We analyzed the phylogenetic and time-space relationships (phylodynamics) of 181 isolates of vesicular stomatitis New Jersey virus (VSNJV) causing disease in Mexico and the United States (US) from 2005 through 2012. We detail the emergence of a genetic lineage in southern Mexico causing outbreaks in central Mexico spreading into northern Mexico and eventually into the US. That emerging lineage showed higher nucleotide sequence identity (99.5%) than that observed for multiple lineages circulating concurrently in southern Mexico (96.8%). Additionally, we identified 58 isolates from Mexico that, unlike previous isolates from Mexico, grouped with northern Central America clade II viruses. This study provides the first direct evidence for the emergence and northward migration of a specific VSNJV genetic lineage from endemic areas in Mexico causing VS outbreaks in the US. In addition we document the emergence of a Central American VSNJV genetic lineage moving northward and causing outbreaks in central Mexico. © 2013 Published by Elsevier Inc.

  19. Faster replication and higher expression levels of viral glycoproteins give the vesicular stomatitis virus/measles virus hybrid VSV-FH a growth advantage over measles virus.

    Science.gov (United States)

    Ayala-Breton, Camilo; Russell, Luke O J; Russell, Stephen J; Peng, Kah-Whye

    2014-08-01

    VSV-FH is a hybrid vesicular stomatitis virus (VSV) with a deletion of its G glycoprotein and encoding the measles virus (MV) fusion (F) and hemagglutinin (H) envelope glycoproteins. VSV-FH infects cells expressing MV receptors and is fusogenic and effective against myeloma xenografts in mice. We evaluated the fusogenic activities of MV and VSV-FH in relationship to the density of receptor on the target cell surface and the kinetics of F and H expression in infected cells. Using a panel of cells expressing increasing numbers of the MV receptor CD46, we evaluated syncytium size in MV- or VSV-FH-infected cells. VSV-FH is not fusogenic at low CD46 density but requires less CD46 for syncytium formation than MV. The size of each syncytium is larger in VSV-FH-infected cells at a specific CD46 density. While syncytium size reached a plateau and did not increase further in MV-infected CHO cells expressing ≥4,620 CD46 copies/cell, there was a corresponding increase in syncytium size with increases in CD46 levels in VSV-FH-infected CD46-expressing CHO (CHO-CD46) cells. Further analysis in VSV-FH-infected cell lines shows earlier and higher expression of F and H mRNAs and protein. However, VSV-FH cytotoxic activity was reduced by pretreatment of the cells with type I interferon. In contrast, the cytopathic effects are not affected in MV-infected cells. In summary, VSV-FH has significant advantages over MV as an oncolytic virus due to its higher viral yield, faster replication kinetics, and larger fusogenic capabilities but should be used in cancer types with defective interferon signaling pathways. We studied the cytotoxic activity of a vesicular stomatitis/measles hybrid virus (VSV-FH), which is superior to that of measles virus (MV), in different cancer cell lines. We determined that viral RNA and protein were produced faster and in higher quantities in VSV-FH-infected cells. This resulted in the formation of larger syncytia, higher production of infectious particles, and

  20. Interferon Beta and Interferon Alpha 2a Differentially Protect Head and Neck Cancer Cells from Vesicular Stomatitis Virus-Induced Oncolysis.

    Science.gov (United States)

    Westcott, Marlena M; Liu, Jingfang; Rajani, Karishma; D'Agostino, Ralph; Lyles, Douglas S; Porosnicu, Mercedes

    2015-08-01

    Oncolytic viruses (OV) preferentially kill cancer cells due in part to defects in their antiviral responses upon exposure to type I interferons (IFNs). However, IFN responsiveness of some tumor cells confers resistance to OV treatment. The human type I IFNs include one IFN-β and multiple IFN-α subtypes that share the same receptor but are capable of differentially inducing biological responses. The role of individual IFN subtypes in promoting tumor cell resistance to OV is addressed here. Two human IFNs which have been produced for clinical use, IFN-α2a and IFN-β, were compared for activity in protecting human head and neck squamous cell carcinoma (HNSCC) lines from oncolysis by vesicular stomatitis virus (VSV). Susceptibility of HNSCC lines to killing by VSV varied. VSV infection induced increased production of IFN-β in resistant HNSCC cells. When added exogenously, IFN-β was significantly more effective at protecting HNSCC cells from VSV oncolysis than was IFN-α2a. In contrast, normal keratinocytes and endothelial cells were protected equivalently by both IFN subtypes. Differential responsiveness of tumor cells to IFN-α and -β was further supported by the finding that autocrine IFN-β but not IFN-α promoted survival of HNSCC cells during persistent VSV infection. Therefore, IFN-α and -β differentially affect VSV oncolysis, justifying the evaluation and comparison of IFN subtypes for use in combination with VSV therapy. Pairing VSV with IFN-α2a may enhance selectivity of oncolytic VSV therapy for HNSCC by inhibiting VSV replication in normal cells without a corresponding inhibition in cancer cells. There has been a great deal of progress in the development of oncolytic viruses. However, a major problem is that individual cancers vary in their sensitivity to oncolytic viruses. In many cases this is due to differences in their production and response to interferons (IFNs). The experiments described here compared the responses of head and neck squamous

  1. Characterization of the mutations responsible for the electrophoretic mobility differences in the NS proteins of vesicular stomatitis virus New Jersey complementation group E mutants.

    Science.gov (United States)

    Rae, B P; Elliott, R M

    1986-12-01

    Temperature-sensitive (ts) mutants of vesicular stomatitis virus, New Jersey serotype, classified in complementation group E contain lesions in the NS gene, which manifest as marked electrophoretic mobility differences of the mutant NS proteins in SDS-polyacrylamide gels. We have cloned full-length cDNA copies of the mutant NS mRNAs, and have determined their nucleotide sequences. tsE1 and tsE3 had single nucleotide changes, and tsE2 had two nucleotide changes, compared to the wild-type NS gene. Three of the mutations were clustered in a region of 18 nucleotides. All the nucleotide differences resulted in amino acid substitutions, which in each case changed the charge of the amino acid concerned. Analysis of the wild-type and mutant NS protein sequences by the method of Chou & Fasman indicated that single amino acid substitutions can radically alter the predicted secondary structure, and these data are discussed in relation to the observed electrophoretic mobility differences.

  2. Interferon regulatory factor-1 protects from fatal neurotropic infection with vesicular stomatitis virus by specific inhibition of viral replication in neurons.

    Directory of Open Access Journals (Sweden)

    Sharmila Nair

    2014-03-01

    Full Text Available The innate immune system protects cells against invading viral pathogens by the auto- and paracrine action of type I interferon (IFN. In addition, the interferon regulatory factor (IRF-1 can induce alternative intrinsic antiviral responses. Although both, type I IFN and IRF-1 mediate their antiviral action by inducing overlapping subsets of IFN stimulated genes, the functional role of this alternative antiviral action of IRF-1 in context of viral infections in vivo remains unknown. Here, we report that IRF-1 is essential to counteract the neuropathology of vesicular stomatitis virus (VSV. IFN- and IRF-1-dependent antiviral responses act sequentially to create a layered antiviral protection program against VSV infections. Upon intranasal infection, VSV is cleared in the presence or absence of IRF-1 in peripheral organs, but IRF-1-/- mice continue to propagate the virus in the brain and succumb. Although rapid IFN induction leads to a decline in VSV titers early on, viral replication is re-enforced in the brains of IRF-1-/- mice. While IFN provides short-term protection, IRF-1 is induced with delayed kinetics and controls viral replication at later stages of infection. IRF-1 has no influence on viral entry but inhibits viral replication in neurons and viral spread through the CNS, which leads to fatal inflammatory responses in the CNS. These data support a temporal, non-redundant antiviral function of type I IFN and IRF-1, the latter playing a crucial role in late time points of VSV infection in the brain.

  3. Vesicular stomatitis virus expressing interferon-β is oncolytic and promotes antitumor immune responses in a syngeneic murine model of non-small cell lung cancer

    Science.gov (United States)

    Patel, Manish R.; Jacobson, Blake A.; Ji, Yan; Drees, Jeremy; Tang, Shaogeng; Xiong, Kerry; Wang, Hengbing; Prigge, Jennifer E.; Dash, Alexander S.; Kratzke, Andrea K.; Mesev, Emily; Etchison, Ryan; Federspiel, Mark J.; Russell, Stephen J.; Kratzke, Robert A.

    2015-01-01

    Vesicular stomatitis virus (VSV) is a potent oncolytic virus for many tumors. VSV that produces interferon-β (VSV-IFNβ) is now in early clinical testing for solid tumors. Here, the preclinical activity of VSV and VSV-IFNβ against non-small cell lung cancer (NSCLC) is reported. NSCLC cell lines were treated in vitro with VSV expressing green fluorescence protein (VSV-GFP) and VSV-IFNβ. VSV-GFP and VSV-IFNβ were active against NSCLC cells. JAK/STAT inhibition with ruxolitinib re-sensitized resistant H838 cells to VSV-IFNβ mediated oncolysis. Intratumoral injections of VSV-GFP and VSV-IFNβ reduced tumor growth and weight in H2009 nude mouse xenografts (p VSV-IFNβ intratumorally. Treatment of LM2 tumors with VSV-IFNβ resulted in tumor regression, prolonged survival (p VSV-IFNβ resulted in decreased tumor-infiltrating regulatory T cells (Treg) and increased CD8+ T cells. Tumor cell expression of PDL-1 was increased after VSV-IFNβ treatment. VSV-IFNβ has potent antitumor effects and promotes systemic antitumor immunity. These data support further clinical investigation of VSV-IFNβ for NSCLC. PMID:26431376

  4. Resistance of human leukocytes to vesicular stomatitis virus infection as one of the innate antiviral immune activities; participation of cell subpopulations.

    Directory of Open Access Journals (Sweden)

    Danuta Duś

    2008-02-01

    Full Text Available Among reactions of innate immunity, resistance of human peripheral blood leukocytes (PBL to viral infection seems important. The purpose of our study was to find, which of the subpopulations of PBL is the most responsible for the innate antiviral immunity of these cells. The innate immunity was measured by using the direct method of infection of leukocytes with vesicular stomatitis virus (VSV. The lack of VSV replication by infected leukocytes (0-1 log TCID50 was taken as an indicator for complete immunity; a low level of VSV (2-3 log for partial immunity; and high VSV titer (more than 4 log for no immunity. The resistance/innate immunity of whole PBL and subpopulations such as: adherent cells, fractions enriched in lymphocytes T, and lymphocytes B (separated on column with nylon wool, NK(+ and NK(- (separated by microbeads activated cell sorting MACS differ from each other. All fractions express higher resistance/innate immunity than the whole PBL. NK(+ cells were found the most resistant fraction of PBL to VSV infection. The results indicate that among the leukocytes in PBL the regulation mechanisms of innate immunity exist. The study on the mechanism of innate immunity regulation as well as the role of NK in innate immunity of PBL must be continued.

  5. Radioiodide imaging and radiovirotherapy of multiple myeloma using VSV(Delta51)-NIS, an attenuated vesicular stomatitis virus encoding the sodium iodide symporter gene.

    Science.gov (United States)

    Goel, Apollina; Carlson, Stephanie K; Classic, Kelly L; Greiner, Suzanne; Naik, Shruthi; Power, Anthony T; Bell, John C; Russell, Stephen J

    2007-10-01

    Multiple myeloma is a radiosensitive malignancy that is currently incurable. Here, we generated a novel recombinant vesicular stomatitis virus [VSV(Delta51)-NIS] that has a deletion of methionine 51 in the matrix protein and expresses the human sodium iodide symporter (NIS) gene. VSV(Delta51)-NIS showed specific oncolytic activity against myeloma cell lines and primary myeloma cells and was able to replicate to high titers in myeloma cells in vitro. Iodide uptake assays showed accumulation of radioactive iodide in VSV(Delta51)-NIS-infected myeloma cells that was specific to the function of the NIS transgene. In bg/nd/xid mice with established subcutaneous myeloma tumors, administration of VSV(Delta51)-NIS resulted in high intratumoral virus replication and tumor regression. VSV-associated neurotoxicity was not observed. Intratumoral spread of the infection was monitored noninvasively by serial gamma camera imaging of (123)I-iodide biodistribution. Dosimetry calculations based on these images pointed to the feasibility of combination radiovirotherapy with VSV(Delta51)-NIS plus (131)I. Immunocompetent mice with syngeneic 5TGM1 myeloma tumors (either subcutaneous or orthotopic) showed significant enhancements of tumor regression and survival when VSV(Delta51)-NIS was combined with (131)I. These results show that VSV(Delta51)-NIS is a safe oncolytic agent with significant therapeutic potential in multiple myeloma.

  6. Cholesterol Supplementation During Production Increases the Infectivity of Retroviral and Lentiviral Vectors Pseudotyped with the Vesicular Stomatitis Virus Glycoprotein (VSV-G).

    Science.gov (United States)

    Chen, Yong; Ott, Christopher J; Townsend, Kay; Subbaiah, Papasani; Aiyar, Ashok; Miller, William M

    2009-05-15

    Cholesterol, a major component of plasma membrane lipid rafts, is important for assembly and budding of enveloped viruses, including influenza and HIV-1. Cholesterol depletion impairs virus assembly and infectivity. This study examined the effects of exogenous cholesterol addition (delivered as a complex with methyl beta cyclodextrin) on the production of Molony murine leukemia virus retroviral vector and HIV-1-based lentiviral vector pseudotyped with the vesicular stomatitis virus glycoprotein (VSV-G). Cholesterol supplementation before and during vector production enhanced the infectivity of retroviral and lentiviral vectors up to 4-fold and 6-fold, respectively. In contrast, the amount of retroviral vector produced was unchanged, and that of lentiviral vector was increased less than two-fold. Both free cholesterol and cholesterol ester content in 293-gag-pol producer cells increased with cholesterol addition. In contrast, the phospholipids headgroup composition was essentially unchanged by cholesterol supplementation in 293-gag-pol packaging cells. Based on these results, it is proposed that cholesterol supplementation increases the infectivity of VSV-G-pseudotyped retroviral and lentiviral vectors, possibly by altering the composition of the producer cell membrane where the viral vectors are assembled and bud, and/or by changing the lipid composition of the viral vectors.

  7. Vesicular stomatitis virus expressing interferon-β is oncolytic and promotes antitumor immune responses in a syngeneic murine model of non-small cell lung cancer.

    Science.gov (United States)

    Patel, Manish R; Jacobson, Blake A; Ji, Yan; Drees, Jeremy; Tang, Shaogeng; Xiong, Kerry; Wang, Hengbing; Prigge, Jennifer E; Dash, Alexander S; Kratzke, Andrea K; Mesev, Emily; Etchison, Ryan; Federspiel, Mark J; Russell, Stephen J; Kratzke, Robert A

    2015-10-20

    Vesicular stomatitis virus (VSV) is a potent oncolytic virus for many tumors. VSV that produces interferon-β (VSV-IFNβ) is now in early clinical testing for solid tumors. Here, the preclinical activity of VSV and VSV-IFNβ against non-small cell lung cancer (NSCLC) is reported. NSCLC cell lines were treated in vitro with VSV expressing green fluorescence protein (VSV-GFP) and VSV-IFNβ. VSV-GFP and VSV-IFNβ were active against NSCLC cells. JAK/STAT inhibition with ruxolitinib re-sensitized resistant H838 cells to VSV-IFNβ mediated oncolysis. Intratumoral injections of VSV-GFP and VSV-IFNβ reduced tumor growth and weight in H2009 nude mouse xenografts (p VSV-IFNβ intratumorally. Treatment of LM2 tumors with VSV-IFNβ resulted in tumor regression, prolonged survival (p VSV-IFNβ resulted in decreased tumor-infiltrating regulatory T cells (Treg) and increased CD8+ T cells. Tumor cell expression of PDL-1 was increased after VSV-IFNβ treatment. VSV-IFNβ has potent antitumor effects and promotes systemic antitumor immunity. These data support further clinical investigation of VSV-IFNβ for NSCLC.

  8. Specific antiviral activity of a poly(L-lysine)-conjugated oligodeoxyribonucleotide sequence complementary to vesicular stomatitis virus N protein mRNA initiation site

    Energy Technology Data Exchange (ETDEWEB)

    Lemaitre, M.; Bayard, B.; Lebleu, B.

    1987-02-01

    Antisense oligonucleotides represent an interesting tool for selective inhibition of gene expression, but their efficient introduction within intact cells provide to be difficult to realize. As a step toward this goal, small (13- or 15-mer) synthetic (/sup 14/C)-oligodeoxyribonucleotides have been coupled at their 3' ends to epsilon-amino groups of lysine residues of poly(L-lysine) (M/sub r/, 14,000). A 15-mer oligonucleotide-poly(L-Lysine) conjugate complementary to the initiation region of vesicular stomatitis virus (VSV) N-protein mRNA specifically inhibits the synthesis of VSV proteins and exerts an antiviral activity against the VSV when added in the cell culture medium at doses as low as 100 nM. Neither synthesis of cellular proteins nor multiplication of encephalomyocarditis virus was affected significantly by this oligonucleotide conjugate. The data suggest that oligonucleotide-poly(L-lysine) conjugates might become effective for studies on gene expression regulation and for antiviral chemotherapy.

  9. Enhanced vesicular stomatitis virus (VSVΔ51 targeting of head and neck cancer in combination with radiation therapy or ZD6126 vascular disrupting agent

    Directory of Open Access Journals (Sweden)

    Alajez Nehad M

    2012-06-01

    Full Text Available Abstract Background Head and neck squamous cell carcinoma (HNSCC is the 5th most common cancer worldwide. Locally advanced HNSCC are treated with either radiation or chemo-radiotherapy, but still associated with high mortality rate, underscoring the need to develop novel therapies. Oncolytic viruses have been garnering increasing interest as anti-cancer agents due to their preferential killing of transformed cells. In this study, we evaluated the therapeutic potential of mutant vesicular stomatitis virus (VSVΔ51 against the human hypopharyngeal FaDu tumour model in vitro and in vivo. Results Our data demonstrated high toxicity of the virus against FaDu cells in vitro, which was associated with induction of apoptosis. In vivo, systemic injection of 1 × 109 pfu had minimal effect on tumour growth; however, when combined with two doses of ionizing radiation (IR; 5 Gy each or a single injection of the vascular disrupting agent (ZD6126, the virus exhibited profound suppression of tumour growth, which translated to a prolonged survival in the treated mice. Concordantly, VSVΔ51 combined with ZD6126 led to a significant increase in viral replication in these tumours. Conclusions Our data suggest that the combinations of VSVΔ51 with either IR or ZD6126 are potentially novel therapeutic opportunities for HNSCC.

  10. N-Terminal Phosphorylation of Phosphoprotein of Vesicular Stomatitis Virus Is Required for Preventing Nucleoprotein from Binding to Cellular RNAs and for Functional Template Formation

    Science.gov (United States)

    Chen, Longyun; Zhang, Shengwei; Banerjee, Amiya K.

    2013-01-01

    The phosphoprotein (P) of vesicular stomatitis virus (VSV) plays essential roles in viral RNA synthesis. It associates with nascent nucleoprotein (N) to form N0-P (free of RNAs), thereby preventing the N from binding to cellular RNAs and maintaining the N in a viral genomic RNA encapsidation-competent form for transcription and replication. The contributions of phosphorylation of P to transcription and replication have been studied intensively, but a concrete mechanism of action still remains unclear. In this study, using a VSV minigenome system, we demonstrated that a mutant of P lacking N-terminal phosphorylation (P3A), in which the N-terminal phosphate acceptor sites are replaced with alanines (S60/A, T62/A, and S64/A), does not support transcription and replication. However, results from protein interaction assays showed that P3A self-associates and interacts with N and the large protein (L) as efficiently as P does. Furthermore, purified recombinant P3A from Sf21 cells supported transcription in an in vitro transcription reconstitution assay. We also proved that P3A is not distributed intranuclearly in vivo. CsCl gradient centrifugation showed that P3A is incapable of preventing N from binding to cellular RNAs and therefore prevents functional template formation. Taken together, our results demonstrate that N-terminal phosphorylation is indispensable for P to prevent N from binding to nonviral RNAs and to maintain the N-specific encapsidation of viral genomic RNA for functional template formation. PMID:23283948

  11. Breaking resistance of pancreatic cancer cells to an attenuated vesicular stomatitis virus through a novel activity of IKK inhibitor TPCA-1.

    Science.gov (United States)

    Cataldi, Marcela; Shah, Nirav R; Felt, Sébastien A; Grdzelishvili, Valery Z

    2015-11-01

    Vesicular stomatitis virus (VSV) is an effective oncolytic virus against most human pancreatic ductal adenocarcinoma (PDAC) cell lines. However, some PDAC cell lines are highly resistant to oncolytic VSV-ΔM51 infection. To better understand the mechanism of resistance, we tested a panel of 16 small molecule inhibitors of different cellular signaling pathways, and identified TPCA-1 (IKK-β inhibitor) and ruxolitinib (JAK1/2 inhibitor), as strong enhancers of VSV-ΔM51 replication and virus-mediated oncolysis in all VSV-resistant PDAC cell lines. Both TPCA-1 and ruxolitinib similarly inhibited STAT1 and STAT2 phosphorylation and decreased expression of antiviral genes MxA and OAS. Moreover, an in situ kinase assay provided biochemical evidence that TPCA-1 directly inhibits JAK1 kinase activity. Together, our data demonstrate that TPCA-1 is a unique dual inhibitor of IKK-β and JAK1 kinase, and provide a new evidence that upregulated type I interferon signaling plays a major role in resistance of pancreatic cancer cells to oncolytic viruses. Copyright © 2015 Elsevier Inc. All rights reserved.

  12. A recombinant vesicular stomatitis virus-based Lassa fever vaccine protects guinea pigs and macaques against challenge with geographically and genetically distinct Lassa viruses.

    Science.gov (United States)

    Safronetz, David; Mire, Chad; Rosenke, Kyle; Feldmann, Friederike; Haddock, Elaine; Geisbert, Thomas; Feldmann, Heinz

    2015-04-01

    Lassa virus (LASV) is endemic in several West African countries and is the etiological agent of Lassa fever. Despite the high annual incidence and significant morbidity and mortality rates, currently there are no approved vaccines to prevent infection or disease in humans. Genetically, LASV demonstrates a high degree of diversity that correlates with geographic distribution. The genetic heterogeneity observed between geographically distinct viruses raises concerns over the potential efficacy of a "universal" LASV vaccine. To date, several experimental LASV vaccines have been developed; however, few have been evaluated against challenge with various genetically unique Lassa virus isolates in relevant animal models. Here we demonstrate that a single, prophylactic immunization with a recombinant vesicular stomatitis virus (VSV) expressing the glycoproteins of LASV strain Josiah from Sierra Leone protects strain 13 guinea pigs from infection / disease following challenge with LASV isolates originating from Liberia, Mali and Nigeria. Similarly, the VSV-based LASV vaccine yields complete protection against a lethal challenge with the Liberian LASV isolate in the gold-standard macaque model of Lassa fever. Our results demonstrate the VSV-based LASV vaccine is capable of preventing morbidity and mortality associated with non-homologous LASV challenge in two animal models of Lassa fever. Additionally, this work highlights the need for the further development of disease models for geographical distinct LASV strains, particularly those from Nigeria, in order to comprehensively evaluate potential vaccines and therapies against this prominent agent of viral hemorrhagic fever.

  13. A recombinant vesicular stomatitis virus-based Lassa fever vaccine protects guinea pigs and macaques against challenge with geographically and genetically distinct Lassa viruses.

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    David Safronetz

    2015-04-01

    Full Text Available Lassa virus (LASV is endemic in several West African countries and is the etiological agent of Lassa fever. Despite the high annual incidence and significant morbidity and mortality rates, currently there are no approved vaccines to prevent infection or disease in humans. Genetically, LASV demonstrates a high degree of diversity that correlates with geographic distribution. The genetic heterogeneity observed between geographically distinct viruses raises concerns over the potential efficacy of a "universal" LASV vaccine. To date, several experimental LASV vaccines have been developed; however, few have been evaluated against challenge with various genetically unique Lassa virus isolates in relevant animal models.Here we demonstrate that a single, prophylactic immunization with a recombinant vesicular stomatitis virus (VSV expressing the glycoproteins of LASV strain Josiah from Sierra Leone protects strain 13 guinea pigs from infection / disease following challenge with LASV isolates originating from Liberia, Mali and Nigeria. Similarly, the VSV-based LASV vaccine yields complete protection against a lethal challenge with the Liberian LASV isolate in the gold-standard macaque model of Lassa fever.Our results demonstrate the VSV-based LASV vaccine is capable of preventing morbidity and mortality associated with non-homologous LASV challenge in two animal models of Lassa fever. Additionally, this work highlights the need for the further development of disease models for geographical distinct LASV strains, particularly those from Nigeria, in order to comprehensively evaluate potential vaccines and therapies against this prominent agent of viral hemorrhagic fever.

  14. Vesicular stomatitis virus replicon expressing the VP2 outer capsid protein of bluetongue virus serotype 8 induces complete protection of sheep against challenge infection.

    Science.gov (United States)

    Kochinger, Stefanie; Renevey, Nathalie; Hofmann, Martin A; Zimmer, Gert

    2014-06-13

    Bluetongue virus (BTV) is an arthropod-borne pathogen that causes an often fatal, hemorrhagic disease in ruminants. Different BTV serotypes occur throughout many temperate and tropical regions of the world. In 2006, BTV serotype 8 (BTV-8) emerged in Central and Northern Europe for the first time. Although this outbreak was eventually controlled using inactivated virus vaccines, the epidemic caused significant economic losses not only from the disease in livestock but also from trade restrictions. To date, BTV vaccines that allow simple serological discrimination of infected and vaccinated animals (DIVA) have not been approved for use in livestock. In this study, we generated recombinant RNA replicon particles based on single-cycle vesicular stomatitis virus (VSV) vectors. Immunization of sheep with infectious VSV replicon particles expressing the outer capsid VP2 protein of BTV-8 resulted in induction of BTV-8 serotype-specific neutralizing antibodies. After challenge with a virulent BTV-8 strain, the vaccinated animals neither developed signs of disease nor showed viremia. In contrast, immunization of sheep with recombinant VP5 - the second outer capsid protein of BTV - did not confer protection. Discrimination of infected from vaccinated animals was readily achieved using an ELISA for detection of antibodies against the VP7 antigen. These data indicate that VSV replicon particles potentially represent a safe and efficacious vaccine platform with which to control future outbreaks by BTV-8 or other serotypes, especially in previously non-endemic regions where discrimination between vaccinated and infected animals is crucial.

  15. Vesicular Stomatitis Virus G Glycoprotein and ATRA Enhanced Bystander Killing of Chemoresistant Leukemic Cells by Herpes Simplex Virus Thymidine Kinase/Ganciclovir.

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    Hu, Chenxi; Chen, Zheng; Zhao, Wenjun; Wei, Lirong; Zheng, Yanwen; He, Chao; Zeng, Yan; Yin, Bin

    2014-02-01

    Refractoriness of acute myeloid leukemia (AML) cells to chemotherapeutics represents a major clinical barrier. Suicide gene therapy for cancer has been attractive but with limited clinical efficacy. In this study, we investigated the potential application of herpes simplex virus thymidine kinase/ganciclovir (HSV-TK/GCV) based system to inhibit chemoresistant AML cells. We first generated Ara-C resistant K562 cells and doxorubicin-resistant THP-1 cells. We found that the HSV-TK/GCV anticancer system suppressed drug resistant leukemic cells in culture. Chemoresistant AML cell lines displayed similar sensitivity to HSV-TK/GCV. Moreover, HSV-TK/GCV killing of leukemic cells was augmented to a mild but significant extent by all-trans retinoic acid (ATRA) with concomitant upregulation of Connexin 43, a major component of gap junctions. Interestingly, HSV-TK/GCV killing was enhanced by expression of vesicular stomatitis virus G glycoprotein (VSV-G), a fusogenic membrane protein, which also increased leukemic cell fusion. Co-culture resistant cells expressing HSV-TK and cells stably transduced with VSV-G showed that expression of VSV-G could promote the bystander killing effect of HSV-TK/GCV. Furthermore, combination of HSV-TK/GCV with VSV-G plus ATRA produced more pronounced antileukemia effect. These results suggest that the HSV-TK/GCV system in combination with fusogenic membrane proteins and/or ATRA could provide a strategy to mitigate the chemoresistance of AML.

  16. Plasma membrane phosphatidylinositol 4,5 bisphosphate is required for internalization of foot-and-mouth disease virus and vesicular stomatitis virus.

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    Angela Vázquez-Calvo

    Full Text Available Phosphatidylinositol-4,5-bisphosphate, PI(4,5P(2, is a phospholipid which plays important roles in clathrin-mediated endocytosis. To investigate the possible role of this lipid on viral entry, two viruses important for animal health were selected: the enveloped vesicular stomatitis virus (VSV - which uses a well characterized clathrin mediated endocytic route - and two different variants of the non-enveloped foot-and-mouth disease virus (FMDV with distinct receptor specificities. The expression of a dominant negative dynamin, a PI(4,5P(2 effector protein, inhibited the internalization and infection of VSV and both FMDV isolates. Depletion of PI(4,5P(2 from plasma membrane using ionomycin or an inducible system, and inhibition of its de novo synthesis with 1-butanol revealed that VSV as well as FMDV C-S8c1, which uses integrins as receptor, displayed a high dependence on PI(4,5P(2 for internalization. Expression of a kinase dead mutant (KD of phosphatidylinositol-4-phosphate-5-kinase Iα (PIP5K-Iα, an enzyme responsible for PI(4,5P(2 synthesis that regulates clathrin-dependent endocytosis, also impaired entry and infection of VSV and FMDV C-S8c1. Interestingly FMDV MARLS variant that uses receptors other than integrins for cell entry was less sensitive to PI(4,5P(2 depletion, and was not inhibited by the expression of the KD PIP5K-Iα mutant suggesting the involvement of endocytic routes other than the clathrin-mediated on its entry. These results highlight the role of PI(4,5P(2 and PIP5K-Iα on clathrin-mediated viral entry.

  17. A vesicular stomatitis virus replicon-based bioassay for the rapid and sensitive determination of multi-species type I interferon.

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    Marianne Berger Rentsch

    Full Text Available Type I interferons (IFN comprise a family of cytokines that signal through a common cellular receptor to induce a plethora of genes with antiviral and other activities. Recombinant IFNs are used for the treatment of hepatitis C virus infection, multiple sclerosis, and certain malignancies. The capability of type I IFN to suppress virus replication and resultant cytopathic effects is frequently used to measure their bioactivity. However, these assays are time-consuming and require appropriate biosafety containment. In this study, an improved IFN assay is presented which is based on a recombinant vesicular stomatitis virus (VSV replicon encoding two reporter proteins, firefly luciferase and green fluorescent protein. The vector lacks the essential envelope glycoprotein (G gene of VSV and is propagated on a G protein-expressing transgenic cell line. Several mammalian and avian cells turned out to be susceptible to infection with the complemented replicon particles. Infected cells readily expressed the reporter proteins at high levels five hours post infection. When human fibroblasts were treated with serial dilutions of human IFN-β prior to infection, reporter expression was accordingly suppressed. This method was more sensitive and faster than a classical IFN bioassay based on VSV cytopathic effects. In addition, the antiviral activity of human IFN-λ (interleukin-29, a type III IFN, was determined on Calu-3 cells. Both IFN-β and IFN-λ were acid-stable, but only IFN-β was resistant to alkaline treatment. The antiviral activities of canine, porcine, and avian type I IFN were analysed with cell lines derived from the corresponding species. This safe bioassay will be useful for the rapid and sensitive quantification of multi-species type I IFN and potentially other antiviral cytokines.

  18. A novel, live-attenuated vesicular stomatitis virus vector displaying conformationally intact, functional HIV-1 envelope trimers that elicits potent cellular and humoral responses in mice.

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    Svetlana Rabinovich

    Full Text Available Though vaccination with live-attenuated SIV provides the greatest protection from progressive disease caused by SIV challenge in rhesus macaques, attenuated HIV presents safety concerns as a vaccine; therefore, live viral vectors carrying HIV immunogens must be considered. We have designed a replication-competent vesicular stomatitis virus (VSV displaying immunogenic HIV-1 Env trimers and attenuating quantities of the native surface glycoprotein (G. The clade B Env immunogen is an Env-VSV G hybrid (EnvG in which the transmembrane and cytoplasmic tail regions are derived from G. Relocation of the G gene to the 5'terminus of the genome and insertion of EnvG into the natural G position induced a ∼1 log reduction in surface G, significant growth attenuation compared to wild-type, and incorporation of abundant EnvG. Western blot analysis indicated that ∼75% of incorporated EnvG was a mature proteolytically processed form. Flow cytometry showed that surface EnvG bound various conformationally- and trimer-specific antibodies (Abs, and in-vitro growth assays on CD4+CCR5+ cells demonstrated EnvG functionality. Neither intranasal (IN or intramuscular (IM administration in mice induced any observable pathology and all regimens tested generated potent Env-specific ELISA titers of 10(4-10(5, with an IM VSV prime/IN VSV boost regimen eliciting the highest binding and neutralizing Ab titers. Significant quantities of Env-specific CD4+ T cells were also detected, which were augmented as much as 70-fold by priming with IM electroporated plasmids encoding EnvG and IL-12. These data suggest that our novel vector can achieve balanced safety and immunogenicity and should be considered as an HIV vaccine platform.

  19. The inhibition of mouse L-cell 45 S ribosomal RNA processing is a highly uv-resistant property of vesicular stomatitis virus

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    Zan, M.; Evans, P.; Lucas-Lenard, J. (Univ. of Connecticut, Storrs (USA))

    1990-07-01

    In mouse L cells infected with vesicular stomatitis virus (VSV), the synthesis of 45 S rRNA and its conversion to 28 S and 18 S rRNA are inhibited during the course of infection. Evidence is presented that the lack of accumulation of stable rRNA species results not only from the decreased transcription and processing of 45 S rRNA, but also from an increased breakdown of pre-rRNA or stable rRNA during processing. In cells prelabeled with (3H)uridine and then infected, the 28 S and 18 S rRNA species remain unaffected. Studies using uv-irradiated VSV indicate that the viral function involved in rRNA synthesis inhibition is slightly more sensitive to uv irradiation than the function involved in processing inhibition. These results suggest that the VSV functions involved in 45 S rRNA synthesis and processing inhibition may be related, or overlapping, but not identical. In cells infected by VSV mutant T1026R1, total RNA synthesis is inhibited, but the distribution of precursor and stable rRNA species remains nearly normal for up to 5 hr after infection. The function of the mutant virus involved in the inhibition of rRNA processing appears to be defective. In mengovirus-infected L cells, 45 S rRNA synthesis, but not processing, is severely inhibited soon after infection, indicating that a decrease in rRNA transcription is not necessarily accompanied by a decrease in processing.

  20. In vivo Biodistribution of a Highly Attenuated Recombinant Vesicular Stomatitis Virus Expressing HIV-1 Gag Following Intramuscular, Intranasal, or Intravenous Inoculation

    Science.gov (United States)

    Johnson, J. Erik; Coleman, John W.; Kalyan, Narender K.; Calderon, Priscilla; Wright, Kevin J.; Obregon, Jennifer; Ogin-Wilson, Eleanor; Natuk, Robert J.; Clarke, David K.; Udem, Stephen A.; Cooper, David; Hendry, R. Michael

    2009-01-01

    Recombinant vesicular stomatitis viruses (rVSVs) are being developed as potential HIV-1 vaccine candidates. To characterize the in vivo replication and dissemination of rVSV vectors in mice, high doses of a highly attenuated vector expressing HIV-1 Gag, rVSVIN- N4CT9-Gag1, and a prototypic reference virus, rVSVIN-HIVGag5, were delivered intramuscularly (IM), intranasally (IN), or intravenously (IV). We used quantitative, real-time RT-PCR (Q-PCR) and standard plaque assays to measure the temporal dissemination of these viruses to various tissues. Following IM inoculation, both viruses were detected primarily at the injection site as well as in draining lymph nodes; neither virus induced significant weight loss, pathologic signs, or evidence of neuroinvasion. In contrast, following IN inoculation, the prototypic virus was detected in all tissues tested and caused significant weight loss leading to death. IN administration of rVSVIN- N4CT9-Gag1 resulted in detection in numerous tissues (brain, lung, nasal turbinates, and lymph nodes) albeit in significantly reduced levels, which caused little or no weight loss nor any mortality. Following IV inoculation, both prototypic and attenuated viruses were detected by Q-PCR in all tissues tested. In contrast to the prototype, rVSVIN-N4CT9-Gag1 viral loads were significantly lower in all organs tested, and no infectious virus was detected in the brain following IV inoculation, despite the presence of viral RNA. These studies demonstrated significant differences in the biodistribution patterns of and the associated pathogenicity engendered by the prototypic and attenuated vectors in a highly susceptible host. PMID:19428903

  1. Attenuation of Recombinant Vesicular Stomatitis Virus-Human Immunodeficiency Virus Type 1 Vaccine Vectors by Gene Translocations and G Gene Truncation Reduces Neurovirulence and Enhances Immunogenicity in Mice▿

    Science.gov (United States)

    Cooper, David; Wright, Kevin J.; Calderon, Priscilla C.; Guo, Min; Nasar, Farooq; Johnson, J. Erik; Coleman, John W.; Lee, Margaret; Kotash, Cheryl; Yurgelonis, Irene; Natuk, Robert J.; Hendry, R. Michael; Udem, Stephen A.; Clarke, David K.

    2008-01-01

    Recombinant vesicular stomatitis virus (rVSV) has shown great potential as a new viral vector for vaccination. However, the prototypic rVSV vector described previously was found to be insufficiently attenuated for clinical evaluation when assessed for neurovirulence in nonhuman primates. Here, we describe the attenuation, neurovirulence, and immunogenicity of rVSV vectors expressing human immunodeficiency virus type 1 Gag. These rVSV vectors were attenuated by combinations of the following manipulations: N gene translocations (N4), G gene truncations (CT1 or CT9), noncytopathic M gene mutations (Mncp), and positioning of the gag gene into the first position of the viral genome (gag1). The resulting N4CT1-gag1, N4CT9-gag1, and MncpCT1-gag1 vectors demonstrated dramatically reduced neurovirulence in mice following direct intracranial inoculation. Surprisingly, in spite of a very high level of attenuation, the N4CT1-gag1 and N4CT9-gag1 vectors generated robust Gag-specific immune responses following intramuscular immunization that were equivalent to or greater than immune responses generated by the more virulent prototypic vectors. MncpCT1-gag1 also induced Gag-specific immune responses following intramuscular immunization that were equivalent to immune responses generated by the prototypic rVSV vector. Placement of the gag gene in the first position of the VSV genome was associated with increased in vitro expression of Gag protein, in vivo expression of Gag mRNA, and enhanced immunogenicity of the vector. These findings demonstrate that through directed manipulation of the rVSV genome, vectors that have reduced neurovirulence and enhanced immunogenicity can be made. PMID:17942549

  2. Live virus vaccines based on a vesicular stomatitis virus (VSV) backbone: Standardized template with key considerations for a risk/benefit assessment.

    Science.gov (United States)

    Clarke, David K; Hendry, R Michael; Singh, Vidisha; Rose, John K; Seligman, Stephen J; Klug, Bettina; Kochhar, Sonali; Mac, Lisa Marie; Carbery, Baevin; Chen, Robert T

    2016-12-12

    The Brighton Collaboration Viral Vector Vaccines Safety Working Group (V3SWG) was formed to evaluate the safety of live, recombinant viral vaccines incorporating genes from heterologous viral and other microbial pathogens in their genome (so-called "chimeric virus vaccines"). Many such viral vector vaccines are now at various stages of clinical evaluation. Here, we introduce an attenuated form of recombinant vesicular stomatitis virus (rVSV) as a potential chimeric virus vaccine for HIV-1, with implications for use as a vaccine vector for other pathogens. The rVSV/HIV-1 vaccine vector was attenuated by combining two major genome modifications. These modifications acted synergistically to greatly enhance vector attenuation and the resulting rVSV vector demonstrated safety in sensitive mouse and non-human primate neurovirulence models. This vector expressing HIV-1 gag protein has completed evaluation in two Phase I clinical trials. In one trial the rVSV/HIV-1 vector was administered in a homologous two-dose regimen, and in a second trial with pDNA in a heterologous prime boost regimen. No serious adverse events were reported nor was vector detected in blood, urine or saliva post vaccination in either trial. Gag specific immune responses were induced in both trials with highest frequency T cell responses detected in the prime boost regimen. The rVSV/HIV-1 vector also demonstrated safety in an ongoing Phase I trial in HIV-1 positive participants. Additionally, clinical trial material has been produced with the rVSV vector expressing HIV-1 env, and Phase I clinical evaluation will initiate in the beginning of 2016. In this paper, we use a standardized template describing key characteristics of the novel rVSV vaccine vectors, in comparison to wild type VSV. The template facilitates scientific discourse among key stakeholders by increasing transparency and comparability of information. The Brighton Collaboration V3SWG template may also be useful as a guide to the

  3. Enhancing the Oncolytic Activity of CD133-Targeted Measles Virus: Receptor Extension or Chimerism with Vesicular Stomatitis Virus Are Most Effective

    Science.gov (United States)

    Kleinlützum, Dina; Hanauer, Julia D. S.; Muik, Alexander; Hanschmann, Kay-Martin; Kays, Sarah-Katharina; Ayala-Breton, Camilo; Peng, Kah-Whye; Mühlebach, Michael D.; Abel, Tobias; Buchholz, Christian J.

    2017-01-01

    Therapy resistance and tumor recurrence are often linked to a small refractory and highly tumorigenic subpopulation of neoplastic cells, known as cancer stem cells (CSCs). A putative marker of CSCs is CD133 (prominin-1). We have previously described a CD133-targeted oncolytic measles virus (MV-CD133) as a promising approach to specifically eliminate CD133-positive tumor cells. Selectivity was introduced at the level of cell entry by an engineered MV hemagglutinin (H). The H protein was blinded for its native receptors and displayed a CD133-specific single-chain antibody fragment (scFv) as targeting domain. Interestingly, MV-CD133 was more active in killing CD133-positive tumors than the unmodified MV-NSe despite being highly selective for its target cells. To further enhance the antitumoral activity of MV-CD133, we here pursued arming technologies, receptor extension, and chimeras between MV-CD133 and vesicular stomatitis virus (VSV). All newly generated viruses including VSV-CD133 were highly selective in eliminating CD133-positive cells. MV-CD46/CD133 killed in addition CD133-negative cells being positive for the MV receptors. In an orthotopic glioma model, MV-CD46/CD133 and MVSCD-CD133, which encodes the super cytosine deaminase, were most effective. Notably, VSV-CD133 caused fatal neurotoxicity in this tumor model. Use of CD133 as receptor could be excluded as being causative. In a subcutaneous tumor model of hepatocellular cancer, VSV-CD133 revealed the most potent oncolytic activity and also significantly prolonged survival of the mice when injected intravenously. Compared to MV-CD133, VSV-CD133 infected a more than 104-fold larger area of the tumor within the same time period. Our data not only suggest new concepts and approaches toward enhancing the oncolytic activity of CD133-targeted oncolytic viruses but also raise awareness about careful toxicity testing of novel virus types. PMID:28695108

  4. Ribose 2'-O methylation of the vesicular stomatitis virus mRNA cap precedes and facilitates subsequent guanine-N-7 methylation by the large polymerase protein.

    Science.gov (United States)

    Rahmeh, Amal A; Li, Jianrong; Kranzusch, Philip J; Whelan, Sean P J

    2009-11-01

    During conventional mRNA cap formation, two separate methyltransferases sequentially modify the cap structure, first at the guanine-N-7 (G-N-7) position and subsequently at the ribose 2'-O position. For vesicular stomatitis virus (VSV), a prototype of the nonsegmented negative-strand RNA viruses, the two methylase activities share a binding site for the methyl donor S-adenosyl-l-methionine and are inhibited by individual amino acid substitutions within the C-terminal domain of the large (L) polymerase protein. This led to the suggestion that a single methylase domain functions for both 2'-O and G-N-7 methylations. Here we report a trans-methylation assay that recapitulates both ribose 2'-O and G-N-7 modifications by using purified recombinant L and in vitro-synthesized RNA. Using this assay, we demonstrate that VSV L typically modifies the 2'-O position of the cap prior to the G-N-7 position and that G-N-7 methylation is diminished by pre-2'-O methylation of the substrate RNA. Amino acid substitutions in the C terminus of L that prevent all cap methylation in recombinant VSV (rVSV) partially retain the ability to G-N-7 methylate a pre-2'-O-methylated RNA, therefore uncoupling the effect of substitutions in the C terminus of the L protein on the two methylations. In addition, we show that the 2'-O and G-N-7 methylase activities act specifically on RNA substrates that contain the conserved elements of a VSV mRNA start at the 5' terminus. This study provides new mechanistic insights into the mRNA cap methylase activities of VSV L, demonstrates that 2'-O methylation precedes and facilitates subsequent G-N-7 methylation, and reveals an RNA sequence and length requirement for the two methylase activities. We propose a model of regulation of the activity of the C terminus of L protein in 2'-O and G-N-7 methylation of the cap structure.

  5. A Confocal and Electron Microscopic Comparison of Interferon β–Induced Changes in Vesicular Stomatitis Virus Infection of Neuroblastoma and Nonneuronal Cells

    Science.gov (United States)

    D'Agostino, Paul M.

    2010-01-01

    Vesicular stomatitis virus (VSV) replication is highly sensitive to interferon (IFN)-induced antiviral responses. Pretreatment of sensitive cultured cells with IFNβ results in a 104-fold reduction in the release of infectious VSV particles. However, differences exist between the mechanisms of reduced infectious particle titers in cell lines of neuroblastoma and nonneuronal lineage. In L929-fibroblast-derived cells, using immunofluorescence confocal microscopy, infection under control conditions reveals the accumulation of VSV matrix, phosphoprotein (P), and nucleocapsid (N) proteins over time, with induced cellular morphological changes indicative of cytopathic effects (CPEs). Upon observing L929 cells that had been pretreated with IFNβ, neither detectable VSV proteins nor CPEs were seen, consistent with type I IFN antiviral protection. When using the same techniques to observe VSV infections of NB41A3 cells, a neuroblastoma cell line, aside from similar viral progression in the untreated control cells, IFNβ-treated cells illustrated a severely attenuated VSV infection. Attenuated VSV progression was observed through detection of VSV matrix, P, and N proteins in isolated cells during the first 8 h of infection. However, by 18–24 h postinfection all neuroblastomas had succumbed to the viral infection. Finally, upon closer inspection of IFNβ-treated NB41A3 cells, no detectable changes in VSV protein localization were identified compared with untreated, virally infected neuroblastomas. Next, to extend our study to test our hypothesis that virion assembly is compromised within type I IFN-treated neuroblastoma cells, we employed electron microscopy to examine our experimental conditions at the ultrastructural level. Using VSV-specific antibodies in conjunction with immuno-gold reagents, we observed several similarities between the two cell lines, such as identification of viroplasmic regions containing VSV N and P proteins and signs of stress-induced CPEs of

  6. Rapid diagnosis of vesicular stomatitis virus in Ecuador by the use of polymerase chain reaction Diagnóstico rápido do vírus da estomatite vesicular no Equador mediante o uso da reação em cadeia da polimerase

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    Lya Madureira Sepúlveda

    2007-09-01

    Full Text Available Vesicular Stomatitis (VS is a viral disease that has a great impact in animal health, as infected animals present marked decrease in meat and milk production. Its presence is a limiting factor for international animal trade. Besides the damage in the livestock productivity, such disease assumes an important role in animal health programs since it is clinically indistinguishable from Foot-and-Mouth Disease. The diagnosis of the VS has been made, mainly, through Complement Fixation, ELISA and Virus Neutralization tests, assays that allow not only for viral detection but also for differentiation of the two serotypes described for Vesicular Stomatitis Virus (VSV: New Jersey (NJ and Indiana (Ind. In this work, a molecular diagnostic approach, the polymerase chain reaction performed after reverse transcription (RT - PCR, based on the specific partial amplification of NS gene of VSV was used, as an alternative method for the detection of the virus. A total of 10 VSV reference samples and 12 specimens collected from animals with clinical signs of vesicular disease obtained from field episodes in Ecuador were tested. The method allowed for the specific partial amplification of the region coding for protein P, both for VSV serotypes New Jersey (642 bp and Indiana 1 (614 bp. The results were compatible with data obtained by Complement Fixation test and the identity of the amplified products was confirmed by nucleotide sequencing.A Estomatite Vesicular (EV é uma enfermidade viral de grande impacto na saúde animal. O animal enfermo apresenta queda na produtividade em rebanho de carne e na produção leiteira, sendo um fator limitante para o comércio internacional de animais. Além dos danos à produtividade essa enfermidade assume importante papel nos programas de saúde animal por ser indistinguível clinicamente da Febre Aftosa. As técnicas para o diagnóstico da EV são, principalmente, a Fixação de Complemento, a ELISA e a Virusneutraliza

  7. Immunogenicity of viral vector, prime-boost SIV vaccine regimens in infant rhesus macaques: attenuated vesicular stomatitis virus (VSV) and modified vaccinia Ankara (MVA) recombinant SIV vaccines compared to live-attenuated SIV.

    Science.gov (United States)

    Van Rompay, Koen K A; Abel, Kristina; Earl, Patricia; Kozlowski, Pamela A; Easlick, Juliet; Moore, Joseph; Buonocore-Buzzelli, Linda; Schmidt, Kimberli A; Wilson, Robert L; Simon, Ian; Moss, Bernard; Rose, Nina; Rose, John; Marthas, Marta L

    2010-02-10

    In a previously developed infant macaque model mimicking HIV infection by breast-feeding, we demonstrated that intramuscular immunization with recombinant poxvirus vaccines expressing simian immunodeficiency virus (SIV) structural proteins provided partial protection against infection following oral inoculation with virulent SIV. In an attempt to further increase systemic but also local antiviral immune responses at the site of viral entry, we tested the immunogenicity of different orally administered, replicating vaccines. One group of newborn macaques received an oral prime immunization with a recombinant vesicular stomatitis virus expressing SIVmac239 gag, pol and env (VSV-SIVgpe), followed 2 weeks later by an intramuscular boost immunization with MVA-SIV. Another group received two immunizations with live-attenuated SIVmac1A11, administered each time both orally and intravenously. Control animals received mock immunizations or non-SIV VSV and MVA control vectors. Analysis of SIV-specific immune responses in blood and lymphoid tissues at 4 weeks of age demonstrated that both vaccine regimens induced systemic antibody responses and both systemic and local cell-mediated immune responses. The safety and immunogenicity of the VSV-SIVgpe+MVA-SIV immunization regimen described in this report provide the scientific incentive to explore the efficacy of this vaccine regimen against virulent SIV exposure in the infant macaque model. Copyright (c) 2009 Elsevier Ltd. All rights reserved.

  8. A single amino acid change resulting in loss of fluorescence of eGFP in a viral fusion protein confers fitness and growth advantage to the recombinant vesicular stomatitis virus

    Science.gov (United States)

    Dinh, Phat X.; Panda, Debasis; Das, Phani B.; Das, Subash C.; Das, Anshuman; Pattnaik, Asit K.

    2012-01-01

    Using a recombinant vesicular stomatitis virus encoding eGFP fused in-frame with an essential viral replication protein, the phosphoprotein P, we show that during passage in culture, the virus mutates the nucleotide C289 within eGFP of the fusion protein PeGFP to A or T, resulting in R97S/C amino acid substitution and loss of fluorescence. The resultant non-fluorescent virus exhibits increased fitness and growth advantage over its fluorescent counterpart. The growth advantage of the non-fluorescent virus appears to be due to increased transcription and replication activities of the PeGFP protein carrying the R97S/C substitution. Further, our results show that the R97S/C mutation occurs prior to accumulation of mutations that can result in loss of expression of the gene inserted at the G-L gene junction. These results suggest that fitness gain is more important for the recombinant virus than elimination of expression of the heterologous gene. PMID:22832124

  9. Translation of mRNAs from vesicular stomatitis virus and vaccinia virus is differentially blocked in cells with depletion of eIF4GI and/or eIF4GII.

    Science.gov (United States)

    Welnowska, Ewelina; Castelló, Alfredo; Moral, Pablo; Carrasco, Luis

    2009-12-04

    Cytolytic viruses abrogate host protein synthesis to maximize the translation of their own mRNAs. In this study, we analyzed the eukaryotic initiation factor (eIF) 4G requirement for translation of vesicular stomatitis virus (VSV) and vaccinia virus (VV) mRNAs in HeLa cells using two different strategies: eIF4G depletion by small interfering RNAs or cleavage of eIF4G by expression of poliovirus 2A protease. Depletion of eIF4GI or eIF4GII moderately inhibits cellular protein synthesis, whereas silencing of both factors has only a slightly higher effect. Under these conditions, the extent of VSV protein synthesis is similar to that of nondepleted control cells, whereas VV expression is substantially reduced. Similar results were obtained when eIF4E was depleted. On the other hand, eIF4G cleavage by poliovirus 2A protease strongly inhibits translation of VV protein expression, whereas translation directed by VSV mRNAs is not abrogated, even though VSV mRNAs are capped. Therefore, the requirement for eIF4F activity is different for VV and VSV, suggesting that the molecular mechanism by which their mRNAs initiate their translation is also different. Consistent with these findings, eIF4GI does not colocalize with ribosomes in VSV-infected cells, while eIF2alpha locates at perinuclear sites coincident with ribosomes.

  10. Lack of Correlation between Virus Barosensitivity and the Presence of a Viral Envelope during Inactivation of Human Rotavirus, Vesicular Stomatitis Virus, and Avian Metapneumovirus by High-Pressure Processing▿

    Science.gov (United States)

    Lou, Fangfei; Neetoo, Hudaa; Li, Junan; Chen, Haiqiang; Li, Jianrong

    2011-01-01

    High-pressure processing (HPP) is a nonthermal technology that has been shown to effectively inactivate a wide range of microorganisms. However, the effectiveness of HPP on inactivation of viruses is relatively less well understood. We systematically investigated the effects of intrinsic (pH) and processing (pressure, time, and temperature) parameters on the pressure inactivation of a nonenveloped virus (human rotavirus [HRV]) and two enveloped viruses (vesicular stomatitis virus [VSV] and avian metapneumovirus [aMPV]). We demonstrated that HPP can efficiently inactivate all tested viruses under optimal conditions, although the pressure susceptibilities and the roles of temperature and pH substantially varied among these viruses regardless of the presence of a viral envelope. We found that VSV was much more stable than most food-borne viruses, whereas aMPV was highly susceptible to HPP. When viruses were held for 2 min under 350 MPa at 4°C, 1.1-log, 3.9-log, and 5.0-log virus reductions were achieved for VSV, HRV, and aMPV, respectively. Both VSV and aMPV were more susceptible to HPP at higher temperature and lower pH. In contrast, HRV was more easily inactivated at higher pH, although temperature did not have a significant impact on inactivation. Furthermore, we demonstrated that the damage of virion structure by disruption of the viral envelope and/or capsid is the primary mechanism underlying HPP-induced viral inactivation. In addition, VSV glycoprotein remained antigenic although VSV was completely inactivated. Taken together, our findings suggest that HPP is a promising technology to eliminate viral contaminants in high-risk foods, water, and other fomites. PMID:22003028

  11. Ribose 2′-O Methylation of the Vesicular Stomatitis Virus mRNA Cap Precedes and Facilitates Subsequent Guanine-N-7 Methylation by the Large Polymerase Protein▿

    Science.gov (United States)

    Rahmeh, Amal A.; Li, Jianrong; Kranzusch, Philip J.; Whelan, Sean P. J.

    2009-01-01

    During conventional mRNA cap formation, two separate methyltransferases sequentially modify the cap structure, first at the guanine-N-7 (G-N-7) position and subsequently at the ribose 2′-O position. For vesicular stomatitis virus (VSV), a prototype of the nonsegmented negative-strand RNA viruses, the two methylase activities share a binding site for the methyl donor S-adenosyl-l-methionine and are inhibited by individual amino acid substitutions within the C-terminal domain of the large (L) polymerase protein. This led to the suggestion that a single methylase domain functions for both 2′-O and G-N-7 methylations. Here we report a trans-methylation assay that recapitulates both ribose 2′-O and G-N-7 modifications by using purified recombinant L and in vitro-synthesized RNA. Using this assay, we demonstrate that VSV L typically modifies the 2′-O position of the cap prior to the G-N-7 position and that G-N-7 methylation is diminished by pre-2′-O methylation of the substrate RNA. Amino acid substitutions in the C terminus of L that prevent all cap methylation in recombinant VSV (rVSV) partially retain the ability to G-N-7 methylate a pre-2′-O-methylated RNA, therefore uncoupling the effect of substitutions in the C terminus of the L protein on the two methylations. In addition, we show that the 2′-O and G-N-7 methylase activities act specifically on RNA substrates that contain the conserved elements of a VSV mRNA start at the 5′ terminus. This study provides new mechanistic insights into the mRNA cap methylase activities of VSV L, demonstrates that 2′-O methylation precedes and facilitates subsequent G-N-7 methylation, and reveals an RNA sequence and length requirement for the two methylase activities. We propose a model of regulation of the activity of the C terminus of L protein in 2′-O and G-N-7 methylation of the cap structure. PMID:19710136

  12. DNA Priming Increases Frequency of T-Cell Responses to a Vesicular Stomatitis Virus HIV Vaccine with Specific Enhancement of CD8+ T-Cell Responses by Interleukin-12 Plasmid DNA.

    Science.gov (United States)

    Li, Shuying S; Kochar, Nidhi K; Elizaga, Marnie; Hay, Christine M; Wilson, Gregory J; Cohen, Kristen W; De Rosa, Stephen C; Xu, Rong; Ota-Setlik, Ayuko; Morris, Daryl; Finak, Greg; Allen, Mary; Tieu, Hong-Van; Frank, Ian; Sobieszczyk, Magdalena E; Hannaman, Drew; Gottardo, Raphael; Gilbert, Peter B; Tomaras, Georgia D; Corey, Lawrence; Clarke, David K; Egan, Michael A; Eldridge, John H; McElrath, M Juliana; Frahm, Nicole

    2017-11-01

    The HIV Vaccine Trials Network (HVTN) 087 vaccine trial assessed the effect of increasing doses of pIL-12 (interleukin-12 delivered as plasmid DNA) adjuvant on the immunogenicity of an HIV-1 multiantigen (MAG) DNA vaccine delivered by electroporation and boosted with a vaccine comprising an attenuated vesicular stomatitis virus expressing HIV-1 Gag (VSV-Gag). We randomized 100 healthy adults to receive placebo or 3 mg HIV-MAG DNA vaccine (ProfectusVax HIV-1 gag/pol or ProfectusVax nef/tat/vif, env) coadministered with pIL-12 at 0, 250, 1,000, or 1,500 μg intramuscularly by electroporation at 0, 1, and 3 months followed by intramuscular inoculation with 3.4 × 107 PFU VSV-Gag vaccine at 6 months. Immune responses were assessed after the prime and boost and 6 months after the last vaccination. High-dose pIL-12 increased the magnitude of CD8+ T-cell responses postboost compared to no pIL-12 (P = 0.02), while CD4+ T-cell responses after the prime were higher in the absence of pIL-12 than with low- and medium-dose pIL-12 (P ≤ 0.05). The VSV boost increased Gag-specific CD4+ and CD8+ T-cell responses in all groups (P T cells), inducing a median of four Gag epitopes in responders. Six to 9 months after the boost, responses decreased in magnitude, but CD8+ T-cell response rates were maintained. The addition of a DNA prime dramatically improved responses to the VSV vaccine tested previously in the HVTN 090 trial, leading to broad epitope targeting and maintained CD8+ T-cell response rates at early memory. The addition of high-dose pIL-12 given with a DNA prime by electroporation and boosted with VSV-Gag increased the CD8+ T-cell responses but decreased the CD4+ responses. This approach may be advantageous in reshaping the T-cell responses to a variety of chronic infections or tumors. (This study has been registered at ClinicalTrials.gov under registration no. NCT01578889.). Copyright © 2017 American Society for Microbiology.

  13. O polietilenoglicol aumenta a penetração do vírus da diarréia viral bovina, do vírus da estomatite vesicular e do vírus sincicial respiratório bovino em células de cultivo Polyethylene glycol increases the penetration of bovine viral diarrhea virus, vesicular stomatitis virus and bovine respiratory syncytial virus in cultured cells

    Directory of Open Access Journals (Sweden)

    Renata Dezengrini

    2009-06-01

    bovine enveloped viruses in culture cells. Penetration efficiency was measured by counting the number of viral plaques produced in bovine kidney cells (MDBK. The addition of 5% PEG (molecular weight 6.000 to the viral inoculum containing 100 TCID50 mL-1 (tissue culture median infectious dosis of each virus, during adsorption for 2h at 37°C, resulted in a significant increase in the number of plaques for bovine viral diarrhea virus (BVDV (increase of 3.4-fold, vesicular stomatitis virus (VSV (2.2-fold and bovine respiratory syncytial virus (BRSV (1.5-fold. The addition of 5% PEG to the inoculum of bovine herpesviruses 1, 2 and 5 (BoHV-1, BoHV-2 and BoHV-5 did not increase the number of viral plaques. On the other hand, PEG produced a reduction in the number of plaques by bovine parainfluenza virus (bPI-3V (1.4-fold. Furthermore, the addition of 5% PEG produced a 10- to 1000-fold increase in the sensitivity of BVDV detection in the serum of three persistently infected calves; and doubled the sensitivity of detection of BRSV in nasal secretions of two experimentally infected sheep. These results demonstrate that PEG enhances the efficiency of infection by BVDV, VSV and BRSV in cultured bovine cells and therefore may be used to increase the sensitivity of virus detection in clinical samples (viral isolation, and/or to increase virus titers in cell cultures.

  14. Effect of proteolytic digestion on the function of vesicular stomatitis ...

    Indian Academy of Sciences (India)

    tribpo

    globulin-G (IgG) followed by antibody to swine-antigoat IgG conjugated to horseradish peroxidase. The proteins were visualized by the color reaction with the substrate 4- chloro-1-napthol. Protein kinase assay. The reaction mixture contained chymotrypsin-treated or untreated N-RNA template. (27 µg), Tris-HCl, pH 8·0, ...

  15. ACUTE STOMATITIS IN CHILDREN

    Directory of Open Access Journals (Sweden)

    L.N. Drobot’ko

    2010-01-01

    Full Text Available The problem of diseases of mucous membrane of oral cavity is one of the main ones in dentistry. Special attention in this problem should be given to the acute herpetic stomatitis. 80% of all cases of mucous membrane of oral cavity in children are herpetic stomatitis. Local immunity in children with acute stomatitis is closely related to the character of course of pathology. An administration of immunomodulatory treatment is pathogenetically grounded. Bacterial lysates mixture causes etiotropical and pathogenetical effect and increases the activity of immune system resulting in relapses prophylaxis.Key words: children, acute herpetic stomatitis, bacterial lysates mixture.(Voprosy sovremennoi pediatrii — Current Pediatrics. 2010;9(2:146-149

  16. Buckwheat stomatal traits under aluminium toxicity

    OpenAIRE

    Oleksandr E. Smirnov; Anatoliy M. Kosyan; Oksana I. Kosyk; Natalia Yu. Taran

    2014-01-01

    Aluminium influence on some stomatal parameters of common buckwheat (Fagopyrum esculentum Moench.) was studied. Significant changes in stomatal density, stomatal index and stomatal shape coefficient under aluminium treatment were revealed. Stomatal closure and no difference in total stomatal potential conductance index of treatment plants were suggested as aluminium resistance characteristics.

  17. Buckwheat stomatal traits under aluminium toxicity

    Directory of Open Access Journals (Sweden)

    Oleksandr E. Smirnov

    2014-04-01

    Full Text Available Aluminium influence on some stomatal parameters of common buckwheat (Fagopyrum esculentum Moench. was studied. Significant changes in stomatal density, stomatal index and stomatal shape coefficient under aluminium treatment were revealed. Stomatal closure and no difference in total stomatal potential conductance index of treatment plants were suggested as aluminium resistance characteristics.

  18. Recurrent Aphthous Stomatitis: A Review.

    Science.gov (United States)

    Edgar, Natalie Rose; Saleh, Dahlia; Miller, Richard A

    2017-03-01

    Aphthous stomatitis is a painful and often recurrent inflammatory process of the oral mucosa that can appear secondary to various well-defined disease processes. Idiopathic recurrent aphthous stomatitis is referred to as recurrent aphthous stomatitis. The differential diagnosis for recurrent aphthous ulcerations is extensive and ranges from idiopathic benign causes to inherited fever syndromes, to connective tissue disease, or even inflammatory bowel diseases. A thorough history and review of systems can assist the clinician in determining whether it is related to a systemic inflammatory process or truly idiopathic. Management of aphthous stomatitis is challenging. For recurrent aphthous stomatitis or recalcitrant aphthous stomatitis from underlying disease, first-line treatment consists of topical medications with use of systemic medications as necessary. Herein, the authors discuss the differential diagnosis and treatment ladder of aphthous stomatitis as described in the literature.

  19. Recurrent aphthous stomatitis.

    Science.gov (United States)

    Akintoye, Sunday O; Greenberg, Martin S

    2005-01-01

    The cause of recurrent aphthous stomatitis (RAS) remains unknown despite considerable research. This article reviews the evidence for current theories regarding this disorder, including possible suspected relationships with microbial and immunologic factors, and presents medical diseases that mimic RAS lesions in certain patients. Topical management of the common form of minor RAS is described along with systemic therapy currently available to patients with severe forms of this disease.

  20. Stomatal Development in Arabidopsis

    Science.gov (United States)

    Pillitteri, Lynn Jo; Dong, Juan

    2013-01-01

    Stomata consist of two guard cells that function as turgor-operated valves that regulate gas exchange in plants. In Arabidopsis, a dedicated cell lineage is initiated and undergoes a series of cell divisions and cell-state transitions to produce a stoma. A set of basic helix-loop-helix (bHLH) transcription factors regulates the transition and differentiation events through the lineage, while the placement of stomata relative to each other is controlled by intercellular signaling via peptide ligands, transmembrane receptors, and mitogen-activated protein kinase (MAPK) modules. Some genes involved in regulating stomatal differentiation or density are also involved in hormonal and environmental stress responses, which may provide a link between modulation of stomatal development or function in response to changes in the environment. Premitotic polarlylocalized proteins provide an added layer of regulation, which can be addressed more thoroughly with the identification of additional proteins in this pathway. Linking the networks that control stomatal development promises to bring advances to our understanding of signal transduction, cell polarity, and cell-fate specification in plants. PMID:23864836

  1. Recurrent Aphthous Stomatitis: A Review

    OpenAIRE

    Edgar, Natalie Rose; Saleh, Dahlia; Miller, Richard A.

    2017-01-01

    Aphthous stomatitis is a painful and often recurrent inflammatory process of the oral mucosa that can appear secondary to various well-defined disease processes. Idiopathic recurrent aphthous stomatitis is referred to as recurrent aphthous stomatitis. The differential diagnosis for recurrent aphthous ulcerations is extensive and ranges from idiopathic benign causes to inherited fever syndromes, to connective tissue disease, or even inflammatory bowel diseases. A thorough history and review of...

  2. [Recurrent aphthous stomatitis in Rheumatology].

    Science.gov (United States)

    Riera Matute, Gabriel; Riera Alonso, Elena

    2011-01-01

    Recurrent aphthous stomatitis consists on recurring oral ulcers of unknown etiology. Oral ulcers may be different in number and size depending on the clinical presentation, which also determines the time needed for healing. Moreover, there are factors associated to outbreaks but not implicated in its etiopathogenesis. When oral aphthosis has a known etiology, it is not considered as recurrent aphthous stomatitis. The severity and the clinical presentation helps in the differential diagnosis. Treatment is symptomatic in recurrent aphthous stomatitis while, if there is an underlying systemic disease, the treatment of such disease is need in addition to topical treatment. Copyright © 2011 Elsevier España, S.L. All rights reserved.

  3. Recurrent aphthous stomatitis.

    Science.gov (United States)

    Cui, Ricky Z; Bruce, Alison J; Rogers, Roy S

    2016-01-01

    Recurrent aphthous stomatitis (RAS) is the most common acute oral ulcerative condition in North America. RAS is divided into a mild, common form, simple aphthosis, and a severe, less common form, complex aphthosis. Aphthosis is a reactive condition. The lesions of RAS can represent the mucosal manifestation of a variety of conditions. These include conditions with oral and genital aphthae such as ulcus vulvae acutum, reactive nonsexually related acute genital ulcers, and Behçet disease. The mouth is the beginning of the gastrointestinal (GI) tract, and the lesions of RAS can be a manifestation of GI diseases such as gluten-sensitive enteropathy, ulcerative colitis, and Crohn disease. Complex aphthosis may also have correctable causes. The clinician should seek these in a careful evaluation. Successful management of both simple and complex aphthosis depends on accurate diagnosis, proper classification, recognition of provocative factors, and the identification of associated diseases. The outlook for patients with both simple and complex aphthosis is positive. Copyright © 2016 Elsevier Inc. All rights reserved.

  4. Effects of stomatal development on stomatal conductance and on stomatal limitation of photosynthesis in Syringa oblata and Euonymus japonicus Thunb.

    Science.gov (United States)

    Wu, Bing-Jie; Chow, Wah Soon; Liu, Yu-Jun; Shi, Lei; Jiang, Chuang-Dao

    2014-12-01

    During leaf development, the increase in stomatal conductance cannot meet photosynthetic demand for CO2, thus leading to stomatal limitation of photosynthesis (Ls). Considering the crucial influences of stomatal development on stomatal conductance, we speculated whether stomatal development limits photosynthesis to some extent. To test this hypothesis, stomatal development, stomatal conductance and photosynthesis were carefully studied in both Syringa oblata (normal greening species) and Euonymus japonicus Thunb (delayed greening species). Our results show that the size of stomata increased gradually with leaf expansion, resulting in increased stomatal conductance up to the time of full leaf expansion. During this process, photosynthesis also increased steadily. Compared to that in S. oblata, the development of chloroplasts in E. japonicus Thunb was obviously delayed, leading to a delay in the improvement of photosynthetic capacity. Further analysis revealed that before full leaf expansion, stomatal limitation increased rapidly in both S. oblata and E. japonicus Thunb; after full leaf expansion, stomatal limitation continually increased in E. japonicus Thunb. Accordingly, we suggested that the enhancement of photosynthetic capacity is the main factor leading to stomatal limitation during leaf development but that stomatal development can alleviate stomatal limitation with the increase of photosynthesis by controlling gas exchange. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  5. A spatio-temporal analysis of matrix protein and nucleocapsid trafficking during vesicular stomatitis virus uncoating.

    Directory of Open Access Journals (Sweden)

    Chad E Mire

    2010-07-01

    Full Text Available To study VSV entry and the fate of incoming matrix (M protein during virus uncoating we used recombinant viruses encoding M proteins with a C-terminal tetracysteine tag that could be fluorescently labeled using biarsenical (Lumio compounds. We found that uncoating occurs early in the endocytic pathway and is inhibited by expression of dominant-negative (DN Rab5, but is not inhibited by DN-Rab7 or DN-Rab11. Uncoating, as defined by the separation of nucleocapsids from M protein, occurred between 15 and 20 minutes post-entry and did not require microtubules or an intact actin cytoskeleton. Unexpectedly, the bulk of M protein remained associated with endosomal membranes after uncoating and was eventually trafficked to recycling endosomes. Another small, but significant fraction of M distributed to nuclear pore complexes, which was also not dependent on microtubules or polymerized actin. Quantification of fluorescence from high-resolution confocal micrographs indicated that after membrane fusion, M protein diffuses across the endosomal membrane with a concomitant increase in fluorescence from the Lumio label which occurred soon after the release of RNPs into the cytoplasm. These data support a new model for VSV uncoating in which RNPs are released from M which remains bound to the endosomal membrane rather than the dissociation of M protein from RNPs after release of the complex into the cytoplasm following membrane fusion.

  6. Methylation of vesicular stomatitis virus (VSV) mRNA 5'-cap structures in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Hammond, D.C.; Lesnaw, J.A.

    1987-05-01

    Monocistronic VSV mRNAs synthesized by subviral particles in vitro display the methylated 5'-cap structure m'G(5')ppp(5')Am. The authors have detected both monomethylated cap structures, m/sup 7/G(5')ppp(5')A and G(5')Am, in reactions containing suboptimal concentrations of AdoMet. To assess the putative precursor roles of these cap structures the authors devised dual label pulse-chase analyses employing S-(CH/sub 3/-/sup 3/H)-AdoMet and (..beta..-/sup 32/P)GTP. The labeled cap structures were analyzed by HPLC. The simultaneous chasing of both radiolabeled substrates allowed 1) the isolation of a specific set of caps labeled as (..beta..-/sup 32/P)-R/sup 7/G(5')ppp(5')AR (R=H or CH/sub 3/) and 2) the determination of the transcriptive fate of each intermediate cap structure within the set. The results demonstrated that both monomethylated cap structures serve as intermediates for the dimethylated cap and that the order of cap methylation is non-compulsory. These data, coupled with previous observations of hypomethylated cap structures in polyadenylated RNAs, have suggested that methylation occurs in a chain length dependent window.

  7. Interactions of macrophages with probiotic bacteria lead to increased antiviral response against vesicular stomatitis virus

    DEFF Research Database (Denmark)

    Ivec, Martin; Botic, Tanja; Koren, Srecko

    2007-01-01

    and by producing chemokines and immunoregulatory cytokines that enable the adaptive immune response to recognize infected cells and perform antiviral effector functions. Probiotics, as a part of the normal gut intestinal flora, are important in supporting a functional yet balanced immune system. Improving our...

  8. Recurrent aphthous stomatitis. An update.

    Science.gov (United States)

    Ship, J A

    1996-02-01

    Recurrent aphthous ulceration or recurrent aphthous stomatitis is the most common oral mucosal disease known to human beings. Despite much clinical and research attention, the causes remain poorly understood, the ulcers are not preventable, and treatment is symptomatic. The most common presentation is minor recurrent aphthous stomatitis: recurrent, round, clearly defined, small, painful ulcers that heal in 10 to 14 days without scarring. Major recurrent aphthous stomatitis lesions are larger (greater than 5 mm), can last for 6 weeks or longer, and frequently scar. The third variety of recurrent aphthous stomatitis is herpetiform ulcers, which present as multiple small clusters of pinpoint lesions that can coalesce to form large irregular ulcers and last 7 to 10 days. Diagnosis of all varieties is usually made after clinical examination. Many local and systemic factors have been associated with these conditions, and there is evidence that there may be a genetic and immunopathogenic basis for recurrent aphthous ulceration. Management of this condition depends on the clinical presentation and symptoms and includes analgesic, antimicrobial, and immunomodulatory drugs. As dental clinicians and researchers become better trained in oral medicine and stomatology, it is anticipated that the pathophysiology, prevention, and treatment of recurrent aphthous ulceration will improve in the future.

  9. Prosthetic stomatitis with removable dentures

    Directory of Open Access Journals (Sweden)

    Rozalieva Yu.Yu.

    2012-06-01

    Full Text Available The Research Objective: To study patients with prosthetic stomatitis, who use the removable laminar dentures. Materials: The consultations and treatment of 79 patients aged 47-65 years have been conducted. The patients have been divided into two clinical groups. The first clinical group (39 persons with the performance of immediate prosthet-ics; the second control clinical group (40 persons — the permanent dentures were produced without the preliminary instruction. Results: All the patients, having the laminar dentures without the preliminary use of immediate constructions of dentures, in spite of repeated correction of them, have had changes of dentures and transitory fold. Patients have been exposed to prosthetic stomatitis of different etiology (without trauma; the single-shot or multiple correction of dentures by the method of rebasing with using of cold cure plastics has been made. Conclusion: Structural and functional changes of dentition during the prosthetic stomatitis lead to disorders, associated by the mucositis. Use of the term of «prosthetic stomatitis» reflects etiological and pathogenetic component of changes in the denture-supporting tissues

  10. Stomatal complex types, stomatal density, and the stomatal index in some species of dioscorea

    Directory of Open Access Journals (Sweden)

    Abdulrahaman A.A.

    2009-01-01

    Full Text Available Dioscorea alata L. has three stomatal complex types, namely, paracytic, anisocytic, and tetracytic stomata, with percentage frequency values of 50, 18, and 32, respectively. Dioscorea bulbifera has paracytic and anisocytic stomata, with percentage frequency values of 87.60 and 12.40, respectively. Dioscorea cayenensis has anisocytic stomata, with a percent­age frequency value of 100. Dioscorea dumetorum has tetracytic and paractytic stomata, with percentage frequency values of 91.05 and 8.95, respectively. Both D. esculenta and D. rotundata have paracytic stomata, with a percentage frequency of 100. The range of variation of stomatal density is from 10 (lowest value in D. alata and D. dumentorum to 27 (highest value in D. bulbifera. The stomatal index also varies, from 24 in D. alata to 47 in D. cayenensis. The size of stomata in all species is small, varying in length from 0.74 μm in D. alata to 1.79 μm in D. dumentorum. An indented dichotomous key based on stomatal features was constructed to distinguish and identify the species.

  11. Allergic Stomatitis From Orthodontic Adhesives.

    Science.gov (United States)

    Peterson, Mark R; Wong, Priscilla H; Dickson, Scott D; Coop, Christopher A

    2017-03-01

    We report a case of a type IV hypersensitivity reaction causing oral stomatitis, presumed to be the result of common dental adhesives. The case was diagnosed using patch testing to the dental adhesives that were used in the patient. Both of the adhesives tested contained a form of acrylate that is being seen more frequently in the literature as a cause of type IV hypersensitivity reactions. Metals can cause allergic reactions; however, other contact items need to be considered as a cause of oral allergic reactions. Cases of allergic stomatitis are rising and there is question if all-in-one adhesives may be contributing to this rise. Reprint & Copyright © 2017 Association of Military Surgeons of the U.S.

  12. An Attempt to Partition Stomatal and Non-stomatal Ozone Deposition Parts on a Short Grassland

    Science.gov (United States)

    Horváth, L.; Koncz, P.; Móring, A.; Nagy, Z.; Pintér, K.; Weidinger, T.

    2017-10-01

    To evaluate the damaging effect of tropospheric ozone on vegetation, it is important to evaluate the stomatal uptake of ozone. Although the stomatal flux is a dominant pathway of ozone deposition onto vegetated surfaces, non-stomatal uptake mechanisms such as soil and cuticular deposition also play a vital role, especially when the leaf area index {LAI}model. We then derived the stomatal conductance of ozone using the Penman-Monteith (PM) theory based on the similarity to water vapour conductance. The non-stomatal conductance was calculated by subtracting the stomatal conductance from the canopy conductance derived from directly-measured fluxes. Our results show that for short vegetation (LAI = 0.25) dry deposition of ozone was dominated by the non-stomatal flux, which exceeded the stomatal flux even during the daytime. At night the stomatal uptake of ozone was found to be negligibly small. In the case of vegetation with {LAI}≈ 1 , the daytime stomatal and non-stomatal fluxes were of the same order of magnitude. These results emphasize that non-stomatal processes must be considered even in the case of well-developed vegetation where cuticular uptake is comparable in magnitude with stomatal uptake, and especially in the case of vegetated surfaces with {LAI}ozone deposition.

  13. Protons Regulate Vesicular Glutamate Transporters through an Allosteric Mechanism.

    Science.gov (United States)

    Eriksen, Jacob; Chang, Roger; McGregor, Matt; Silm, Katlin; Suzuki, Toshiharu; Edwards, Robert H

    2016-05-18

    The quantal nature of synaptic transmission requires a mechanism to transport neurotransmitter into synaptic vesicles without promoting non-vesicular efflux across the plasma membrane. Indeed, the vesicular transport of most classical transmitters involves a mechanism of H(+) exchange, which restricts flux to acidic membranes such as synaptic vesicles. However, vesicular transport of the principal excitatory transmitter glutamate depends primarily on membrane potential, which would drive non-vesicular efflux, and the role of protons is unclear. Adapting electrophysiology to record currents associated with the vesicular glutamate transporters (VGLUTs), we characterize a chloride conductance that is gated by lumenal protons and chloride and supports glutamate uptake. Rather than coupling stoichiometrically to glutamate flux, lumenal protons and chloride allosterically activate vesicular glutamate transport. Gating by protons serves to inhibit what would otherwise be substantial non-vesicular glutamate efflux at the plasma membrane, thereby restricting VGLUT activity to synaptic vesicles. Copyright © 2016 Elsevier Inc. All rights reserved.

  14. Microtubule arrays and Arabidopsis stomatal development

    National Research Council Canada - National Science Library

    Jessica R. Lucas; Jeanette A. Nadeau; Fred D. Sack

    Microtubule arrays in living cells were analysed during Arabidopsis stomatal development in order to more closely define stages in the pathway and contexts where intercellular signalling might operate...

  15. Identification of a vesicular aspartate transporter

    OpenAIRE

    Miyaji, Takaaki; Echigo, Noriko; Hiasa, Miki; Senoh, Shigenori; Omote, Hiroshi; Moriyama, Yoshinori

    2008-01-01

    Aspartate is an excitatory amino acid that is costored with glutamate in synaptic vesicles of hippocampal neurons and synaptic-like microvesicles (SLMVs) of pinealocytes and is exocytosed and stimulates neighboring cells by binding to specific cell receptors. Although evidence increasingly supports the occurrence of aspartergic neurotransmission, this process is still debated because the mechanism for the vesicular storage of aspartate is unknown. Here, we show that sialin, a lysosomal H+/sia...

  16. Optimal stomatal behaviour around the world

    DEFF Research Database (Denmark)

    Lin, Yan-Shih; Medlyn, Belinda E.; Duursma, Remko A.

    2015-01-01

    , a globalscale database and an associated globally applicable model of gs that allow predictions of stomatal behaviour are lacking. Here,we present a database of globally distributed gs obtained in the field for a wide range of plant functional types (PFTs) and biomes. We find that stomatal behaviour diers among...

  17. Urban legends: recurrent aphthous stomatitis.

    Science.gov (United States)

    Baccaglini, L; Lalla, R V; Bruce, A J; Sartori-Valinotti, J C; Latortue, M C; Carrozzo, M; Rogers, R S

    2011-11-01

    Recurrent aphthous stomatitis (RAS) is the most common idiopathic intraoral ulcerative disease in the USA. Aphthae typically occur in apparently healthy individuals, although an association with certain systemic diseases has been reported. Despite the unclear etiopathogenesis, new drug trials are continuously conducted in an attempt to reduce pain and dysfunction. We investigated four controversial topics: (1) Is complex aphthosis a mild form of Behçet's disease (BD)? (2) Is periodic fever, aphthous stomatitis, pharyngitis, and adenitis (PFAPA) syndrome a distinct medical entity? (3) Is RAS associated with other systemic diseases [e.g., celiac disease (CD) and B12 deficiency]? (4) Are there any new RAS treatments? Results from extensive literature searches, including a systematic review of RAS trials, suggested the following: (1) Complex aphthosis is not a mild form of BD in North America or Western Europe; (2) Diagnostic criteria for PFAPA have low specificity and the characteristics of the oral ulcers warrant further studies; (3) Oral ulcers may be associated with CD; however, these ulcers may not be RAS; RAS is rarely associated with B12 deficiency; nevertheless, B12 treatment may be beneficial, via mechanisms that warrant further study; (4) Thirty-three controlled trials published in the past 6 years reported some effectiveness, although potential for bias was high. © 2011 John Wiley & Sons A/S.

  18. Histopathological study of stomatitis nicotina.

    Science.gov (United States)

    Reddy, C R; Kameswari, V R; Ramulu, C; Reddy, P G

    1971-09-01

    One hundred and thirteen biopsies of the palate in people accustomed to smoking cigars, most of them with the burning end of the cigar inside the mouth, have been studied.Thirty-eight of these showed mild to severe atypical changes in the epithelium. There were 19 lesions showing orthokeratosis and 53 showing hyperorthokeratosis.The earliest atypical change is seen in the mouths of the ducts of the glands.There were 3 cases showing microinvasive carcinomas.Pigmentation is a prominent feature in these cases.The papules with umbilication could be due to hyperplasia of the mucous glands.It is suggested that stomatitis nicotina occurring in men and women with the habit of reverse smoking is probably precancerous because of the presence of atypical changes in the epithelium and also the finding of 3 microinvasive carcinomas without any macroscopic evidence.There is no acceptable explanation why the soft palate escapes getting either stomatitis nicotina lesion or carcinoma in reverse smokers.

  19. Stomatal conductance increases with rising temperature.

    Science.gov (United States)

    Urban, Josef; Ingwers, Miles; McGuire, Mary Anne; Teskey, Robert O

    2017-08-03

    Stomatal conductance directly modifies plant water relations and photosynthesis. Many environmental factors affecting the stomatal conductance have been intensively studied but temperature has been largely neglected, even though it is one of the fastest changing environmental variables and it is rising due to climate change. In this study, we describe how stomata open when the temperature increases. Stomatal conductance increased by ca 40% in a broadleaf and a coniferous species, poplar (Populus deltoides x nigra) and loblolly pine (Pinus taeda) when temperature was increased by 10 °C, from 30 °C to 40 °C at a constant vapor pressure deficit of 1 kPa. The mechanism of regulating stomatal conductance by temperature was, at least partly, independent of other known mechanisms linked to water status and carbon metabolism. Stomatal conductance increased with rising temperature despite the decrease in leaf water potential, increase in transpiration, increase in intercellular CO2 concentration and was decoupled from photosynthesis. Increase in xylem and mesophyll hydraulic conductance coming from lower water viscosity may to some degree explain temperature dependent opening of stomata. The direct stomatal response to temperature allows plants to benefit from increased evaporative cooling during the heat waves and from lower stomatal limitations to photosynthesis but they may be jeopardized by faster depletion of soil water.

  20. Identification of a vesicular aspartate transporter

    Science.gov (United States)

    Miyaji, Takaaki; Echigo, Noriko; Hiasa, Miki; Senoh, Shigenori; Omote, Hiroshi; Moriyama, Yoshinori

    2008-01-01

    Aspartate is an excitatory amino acid that is costored with glutamate in synaptic vesicles of hippocampal neurons and synaptic-like microvesicles (SLMVs) of pinealocytes and is exocytosed and stimulates neighboring cells by binding to specific cell receptors. Although evidence increasingly supports the occurrence of aspartergic neurotransmission, this process is still debated because the mechanism for the vesicular storage of aspartate is unknown. Here, we show that sialin, a lysosomal H+/sialic acid cotransporter, is present in hippocampal synaptic vesicles and pineal SLMVs. RNA interference of sialin expression decreased exocytosis of aspartate and glutamate in pinealocytes. Proteoliposomes containing purified sialin actively accumulated aspartate and glutamate to a similar extent when inside positive membrane potential is imposed as the driving force. Sialin carrying a mutation found in people suffering from Salla disease (R39C) was completely devoid of aspartate and glutamate transport activity, although it retained appreciable H+/sialic acid cotransport activity. These results strongly suggest that sialin possesses dual physiological functions and acts as a vesicular aspartate/glutamate transporter. It is possible that people with Salla disease lose aspartergic (and also the associated glutamatergic) neurotransmission, and this could provide an explanation for why Salla disease causes severe neurological defects. PMID:18695252

  1. TREATMENT OF APHTHOUS STOMATITIS IN ADOLESCENTS

    National Research Council Canada - National Science Library

    L. M. Kozlova; O. A. Zorina; N. B. Petrukhina

    2014-01-01

    ...) has been in use in clinical practice for years. The study was aimed at comparing the clinical efficacy of the drug based on the listed active substances and other stomatological agents in adolescents with aphthous stomatitis. Results...

  2. Relating Stomatal Conductance to Leaf Functional Traits.

    Science.gov (United States)

    Kröber, Wenzel; Plath, Isa; Heklau, Heike; Bruelheide, Helge

    2015-10-12

    Leaf functional traits are important because they reflect physiological functions, such as transpiration and carbon assimilation. In particular, morphological leaf traits have the potential to summarize plants strategies in terms of water use efficiency, growth pattern and nutrient use. The leaf economics spectrum (LES) is a recognized framework in functional plant ecology and reflects a gradient of increasing specific leaf area (SLA), leaf nitrogen, phosphorus and cation content, and decreasing leaf dry matter content (LDMC) and carbon nitrogen ratio (CN). The LES describes different strategies ranging from that of short-lived leaves with high photosynthetic capacity per leaf mass to long-lived leaves with low mass-based carbon assimilation rates. However, traits that are not included in the LES might provide additional information on the species' physiology, such as those related to stomatal control. Protocols are presented for a wide range of leaf functional traits, including traits of the LES, but also traits that are independent of the LES. In particular, a new method is introduced that relates the plants' regulatory behavior in stomatal conductance to vapor pressure deficit. The resulting parameters of stomatal regulation can then be compared to the LES and other plant functional traits. The results show that functional leaf traits of the LES were also valid predictors for the parameters of stomatal regulation. For example, leaf carbon concentration was positively related to the vapor pressure deficit (vpd) at the point of inflection and the maximum of the conductance-vpd curve. However, traits that are not included in the LES added information in explaining parameters of stomatal control: the vpd at the point of inflection of the conductance-vpd curve was lower for species with higher stomatal density and higher stomatal index. Overall, stomata and vein traits were more powerful predictors for explaining stomatal regulation than traits used in the LES.

  3. AIDS and Recurrent Aphthous Stomatitis.

    Science.gov (United States)

    Miziara, Ivan Dieb; Araujo Filho, Bernardo Cunha; Weber, Raimar

    2005-01-01

    The immunodeficiency state in HIV infected patients has been the cause of severe episodes of Recurrent Aphthous Stomatitis (RAS). Our study aims to establish correlation between the manifestations of RAS and the immunosuppression state caused by HIV infection, through counting of CD4+ cells, CD8+ cells, CD4+:CD8+ cells ratio and viral load. Series study. Ninety-four HIV infected patients (25 women and 69 men) with RAS were evaluated in the ENT Department of the University of Sao Paulo-Medical School from January 1998 to December 2003. The age ranged between 19 and 63 years (mean = 35.3 years). The patients were divided in two groups: AIDS group and HIV infected group. The patients with AIDS and HIV infection presented, respectively, eight ulcers and two ulcers by outbreaks. Similarly, patients with major RAS presented smaller counting of cells CD8+, CD4+ and CD4+/CD8+ cells, and higher mean value of viral load than the patients with herpetiform and minor RAS. Between patients with minor and herpetiform RAS there were no statistical differences. The emergence of the lesions, mainly in major RAS, is directly related to the immunological state of the HIV infected patient. These patients frequently present nutritional deficits and worsening in life style. Thus, diagnosis and treatment of RAS is a challenge that should not be neglected.

  4. Role of Intermediate Filaments in Vesicular Traffic

    Directory of Open Access Journals (Sweden)

    Azzurra Margiotta

    2016-04-01

    Full Text Available Intermediate filaments are an important component of the cellular cytoskeleton. The first established role attributed to intermediate filaments was the mechanical support to cells. However, it is now clear that intermediate filaments have many different roles affecting a variety of other biological functions, such as the organization of microtubules and microfilaments, the regulation of nuclear structure and activity, the control of cell cycle and the regulation of signal transduction pathways. Furthermore, a number of intermediate filament proteins have been involved in the acquisition of tumorigenic properties. Over the last years, a strong involvement of intermediate filament proteins in the regulation of several aspects of intracellular trafficking has strongly emerged. Here, we review the functions of intermediate filaments proteins focusing mainly on the recent knowledge gained from the discovery that intermediate filaments associate with key proteins of the vesicular membrane transport machinery. In particular, we analyze the current understanding of the contribution of intermediate filaments to the endocytic pathway.

  5. [Stomatitis in childhood, not always benign].

    Science.gov (United States)

    Oudshoorn, A M; Ramaker, C

    2000-10-14

    Two boys of 1 and 16 year had painful buccal lesions and were admitted for dehydration. The younger had finger and toe blisters; the older, severely ill, had conjunctivitis, urethritis and skin lesions. Only symptomatic treatment with lidocaine gel and paracetamol gave good recovery. A 5-year-old Turkish girl had recurrent painful buccal ulcers which each time cleared up spontaneously. Stomatitis is common in childhood. Viral infections are the most common causes of stomatitis, in particular infections with herpes simplex virus (herpes gingivostomatitis), Coxsackie virus (herpangina, hand-foot-mouth-disease), chickenpox and infectious mononucleosis. Bacterial infections are rare and mostly secondary to the viral infections. In infants oral candidiasis (thrush) is a common cause of stomatitis. Most infections are self-limiting and reassurance of parents is important. Dehydration is a common complication and admission to hospital can be prevented by analgesics. The most important non-infectious conditions that cause stomatitis in children are recurrent aphthous stomatitis, erythema multiforme major (Stevens-Johnson syndrome), Behçet's disease, malignancy (leukaemia), immune-mediated disorders (agranulocytosis, cyclic neutropenia), traumata, blistering disorders of the skin and lichen planus. A complete history and a thorough physical examination usually give the correct diagnosis and further investigations are seldom necessary.

  6. Stomatal and non-stomatal factors regulated the photosynthesis of soybean seedlings in the present of exogenous bisphenol A.

    Science.gov (United States)

    Jiao, Liya; Wang, Lihong; Zhou, Qing; Huang, Xiaohua

    2017-11-01

    Bisphenol A (BPA) is an emerging environmental endocrine disruptor that has toxic effects on plants growth. Photosynthesis supplies the substances and energy required for plant growth, and regulated by stomatal and non-stomatal factors. Therefore, in this study, to reveal how BPA affects photosynthesis in soybean seedlings (Glycine max L.) from the perspective of stomatal and non-stomatal factors, the stomatal factors (stomatal conductance and behaviours) and non-stomatal factors (Hill reaction, apparent quantum efficiency, Rubisco activity, carboxylation efficiency, the maximum Rubisco carboxylation velocity, ribulose-1,5-bisphospate regeneration capacities mediated by maximum electron transport rates, and triose phosphate utilization rate) were investigated using a portable photosynthesis system. Moreover, the pollution of BPA in the environment was simulated. The results indicate that low-dose BPA enhanced net photosynthetic rate (Pn) primarily by promoting stomatal factors, resulting in increased relative growth rates and accelerated soybean seedling growth. High-dose BPA decreases the Pn by simultaneously inhibiting stomatal and non-stomatal factors, and this inhibition decreases the relative growth rates further reducing soybean seedling growth. Following the withdrawal of BPA, all of the indices were restored to varying degrees. In conclusion, low-dose BPA increased the Pn by promoting stomatal factors while high-dose BPA decreased the Pn by simultaneously inhibiting stomatal and non-stomatal factors. These findings provide a model (or, hypothesis) for the effects of BPA on plant photosynthesis. Copyright © 2017 Elsevier Inc. All rights reserved.

  7. Vesicular secretion of auxin: Evidences and implications.

    Science.gov (United States)

    Baluska, Frantisek; Schlicht, Markus; Volkmann, Dieter; Mancuso, Stefano

    2008-04-01

    The plant hormone auxin is secreted in root apices via phospholipase Dzeta2 (PLDzeta2) activity which produces specific population of phosphatidic acid that stimulates secretion of vesicles enriched with auxin. These vesicles were reported to be localized at plant synapses which are active in auxin secretion, especially at the transition zone of the root apex. There are several implications of this vesicular secretion of auxin. In root apices, auxin emerges as plant neurotransmitter-like signal molecule which coordinates activities of adjacent cells via electric and chemical signaling. Putative quantal release of auxin after electrical stimulation, if confirmed, would be part of neuronal communication between plant cells. As auxin transport across plant synapses is tightly linked with integrated sensory perception of environment, especially of omnipresent gravity and light, this process is proposed to mediate the plant perception of environment. These neuronal features allow sessile plants to integrate multitude of sensory signals into the adaptive behavior of whole plants and the animal-like exploratory behavior of growing roots.

  8. Tratamiento actual de la litiasis vesicular Current treatment of vesicular lithiasis

    Directory of Open Access Journals (Sweden)

    Oscar García Rodríguez

    2010-06-01

    Full Text Available El tratamiento quirúrgico de la litiasis vesicular ha cambiado en los últimos años. La incorporación de las nuevas conductas en la práctica médica diaria no siempre es inmediata. Se argumentan las razones relativas a cuándo operar a un paciente con cálculos en la vesícula biliar, y se documenta cómo este procedimiento se reserva fundamentalmente para los pacientes sintomáticos, considerando el dolor como el síntoma por excelencia. También se expone cómo se ha enfrentado este cambio.Surgical treatment of vesicular lithiasis has changed in past years. The addition of the new techniques in daily medical practice not always is immediate. Reasons relative to when to operate a patient presenting with gall bladder calculi are argued and documenting how this procedure is mainly reserved for symptomatic patients where pain is considered as a symptom par excellence . Also, it is exposed how this change has been faced.

  9. In Vivo Replication and Pathogenesis of Vesicular Stomatitis Virus Recombinant M40 Containing Ebola Virus L-Domain Sequences

    Directory of Open Access Journals (Sweden)

    Takashi Irie

    2012-01-01

    Full Text Available The M40 VSV recombinant was engineered to contain overlapping PTAP and PPxY L-domain motifs and flanking residues from the VP40 protein of Ebola virus. Replication of M40 in cell culture is virtually indistinguishable from that of control viruses. However, the presence of the Ebola PTAP motif in the M40 recombinant enabled this virus to interact with and recruit host Tsg101, which was packaged into M40 virions. In this brief report, we compared replication and the pathogenic profiles of M40 and the parental virus M51R in mice to determine whether the presence of the Ebola L-domains and flanking residues altered in vivo characteristics of the virus. Overall, the in vivo characteristics of M40 were similar to those of the parental M51R virus, indicating that the Ebola sequences did not alter pathogenesis of VSV in this small animal model of infection.

  10. Interferon induction by viruses. VIII. Vesicular stomatitis virus: (+-)DI-011 particles induce interferon in the absence of standard virions

    Energy Technology Data Exchange (ETDEWEB)

    Sekellick, M.J.; Marcus, P.I.

    1982-02-01

    Evidence was presented that VSV (+-)DI-011 particles, which contain a genome of covalently linked totally self-complementary RNA, were excellent inducers of interferon (IFN) - by virtue of the dsRNA presumed to form within an infected cell, and that one molecule of that dsRNA per cell sufficed to induce a quantum yield of IFN. While the IFN-inducing capacity of (+-)DI-011 particle preparations has been confirmed, some researchers contend that DIP by themselves cannot induce IFN and that induction requires the presence of coinfecting (contaminating) standard VSV PFP. Consequently, we reexamined this question and now report that under five different conditions where the function of contaminating standard virus is reduced markedly, or eliminated, there was no diminution of the interferon-inducing particle (IFP) activity in preparations of (+-)DI-011 particles. Thus, inactivation of contaminating PFP by uv radiation or heat, the elimination (during induction) of cycling infection through the use of anti-serum (in the case of mouse L cells), and the reduction of PFP by four successive velocity sedimentation-gradient purifications had no adverse affect on the IFN-inducing capacity of DI-011 in either ''aged'' primary chick embryo cells or in mouse L(Y) cells. Furthermore, dilutions of DI-011 stocks which precluded the presence of even a single PFP still induced IFN in ''aged'' chick embryo cells. In concert these data demonstrate convincingly that IFN induction by DI-011 particles does not require coinfection with standard virus. It follows that DI-011 particles are intrinsically capable of inducing IFN.

  11. In silico and in vitro studies of cytotoxic activity of different peptides derived from vesicular stomatitis virus G protein

    Directory of Open Access Journals (Sweden)

    Fereshte Ghandehari

    2015-01-01

    Conclusion: The results confirmed that P26 and P7 peptides might induce membrane damage and initiate apoptosis. The present study suggested that P26 and P7 peptides could be appropriate candidates for further studies as cytotoxic agents and modifications in the residue at positions 70-280 might potentially produce a more efficient VSVG protein in gene therapy.

  12. Inactivation of a Human Norovirus Surrogate, Human Norovirus Virus-Like Particles, and Vesicular Stomatitis Virus by Gamma Irradiation ▿

    OpenAIRE

    Feng, Kurtis; Divers, Erin; Ma, Yuanmei; Li, Jianrong

    2011-01-01

    Gamma irradiation is a nonthermal processing technology that has been used for the preservation of a variety of food products. This technology has been shown to effectively inactivate bacterial pathogens. Currently, the FDA has approved doses of up to 4.0 kGy to control food-borne pathogens in fresh iceberg lettuce and spinach. However, whether this dose range effectively inactivates food-borne viruses is less understood. We have performed a systematic study on the inactivation of a human nor...

  13. Optimal stomatal behaviour around the world

    Science.gov (United States)

    Lin, Yan-Shih; Medlyn, Belinda E.; Duursma, Remko A.; Prentice, I. Colin; Wang, Han; Baig, Sofia; Eamus, Derek; de Dios, Victor Resco; Mitchell, Patrick; Ellsworth, David S.; de Beeck, Maarten Op; Wallin, Göran; Uddling, Johan; Tarvainen, Lasse; Linderson, Maj-Lena; Cernusak, Lucas A.; Nippert, Jesse B.; Ocheltree, Troy W.; Tissue, David T.; Martin-Stpaul, Nicolas K.; Rogers, Alistair; Warren, Jeff M.; de Angelis, Paolo; Hikosaka, Kouki; Han, Qingmin; Onoda, Yusuke; Gimeno, Teresa E.; Barton, Craig V. M.; Bennie, Jonathan; Bonal, Damien; Bosc, Alexandre; Löw, Markus; Macinins-Ng, Cate; Rey, Ana; Rowland, Lucy; Setterfield, Samantha A.; Tausz-Posch, Sabine; Zaragoza-Castells, Joana; Broadmeadow, Mark S. J.; Drake, John E.; Freeman, Michael; Ghannoum, Oula; Hutley, Lindsay B.; Kelly, Jeff W.; Kikuzawa, Kihachiro; Kolari, Pasi; Koyama, Kohei; Limousin, Jean-Marc; Meir, Patrick; Lola da Costa, Antonio C.; Mikkelsen, Teis N.; Salinas, Norma; Sun, Wei; Wingate, Lisa

    2015-05-01

    Stomatal conductance (gs) is a key land-surface attribute as it links transpiration, the dominant component of global land evapotranspiration, and photosynthesis, the driving force of the global carbon cycle. Despite the pivotal role of gs in predictions of global water and carbon cycle changes, a global-scale database and an associated globally applicable model of gs that allow predictions of stomatal behaviour are lacking. Here, we present a database of globally distributed gs obtained in the field for a wide range of plant functional types (PFTs) and biomes. We find that stomatal behaviour differs among PFTs according to their marginal carbon cost of water use, as predicted by the theory underpinning the optimal stomatal model and the leaf and wood economics spectrum. We also demonstrate a global relationship with climate. These findings provide a robust theoretical framework for understanding and predicting the behaviour of gs across biomes and across PFTs that can be applied to regional, continental and global-scale modelling of ecosystem productivity, energy balance and ecohydrological processes in a future changing climate.

  14. Brassinosteroids modulate ABA-induced stomatal closure in Arabidopsis.

    Science.gov (United States)

    Ha, Yunmi; Shang, Yun; Nam, Kyoung Hee

    2016-12-01

    Stomatal movement in response to water availability is an important physiological process in the survival of land plants. The plant hormone abscisic acid (ABA) and brassinosteroids (BRs) regulate stomatal closure. The physiological functions of ABA and BRs, including germination, cell elongation and stomatal movement, are generally known to be antagonistic. Here, we investigated how BRs affect stomatal movement alone and in combination with ABA. We demonstrate that brassinoslide (BL), the most active BR, promotes stomatal closure in an ABA-independent manner. Interestingly, BL also inhibited ABA-induced stomatal closure when a high concentration of BL was added to ABA. Furthermore, we found that the induction of some genes for reactive oxygen species (ROS) generation by ABA (AtrbohD, NIA1 and NIA2) and subsequent ROS production were repressed by BL treatment. The BR signaling mutant bri1-301 failed to inhibit ABA-induced stomatal closure upon BL treatment. However, BRI1-overexpressing transgenic plants were hypersensitive to ABA during stomatal closure, and BL reversed ABA-induced stomatal closure more completely than in wild type plants. Taken together, these results suggest that BRs can positively and negatively modulate ABA-induced stomatal closure. Therefore, interactions between ABA and BR signaling are important for the regulation of stomatal closure. © The Author 2016. Published by Oxford University Press on behalf of the Society for Experimental Biology.

  15. Tratamiento actual de la litiasis vesicular Current treatment of vesicular lithiasis

    OpenAIRE

    Oscar García Rodríguez

    2010-01-01

    El tratamiento quirúrgico de la litiasis vesicular ha cambiado en los últimos años. La incorporación de las nuevas conductas en la práctica médica diaria no siempre es inmediata. Se argumentan las razones relativas a cuándo operar a un paciente con cálculos en la vesícula biliar, y se documenta cómo este procedimiento se reserva fundamentalmente para los pacientes sintomáticos, considerando el dolor como el síntoma por excelencia. También se expone cómo se ha enfrentado este cambio.Surgical t...

  16. Molecular physiology of vesicular glutamate transporters in the digestive system

    Institute of Scientific and Technical Information of China (English)

    Tao Li; Fayez K. Ghishan; Liqun Bai

    2005-01-01

    Glutamate is the major excitatory neurotransmitter in the mammalian central nervous system (CNS). Packaging and storage of glutamate into glutamatergic neuronal vesicles require ATP-dependent vesicular glutamate uptake systems, which utilize the electrochemical proton gradient as a driving force. Three vesicular glutamate transporters (VGLUT1-3) have been recently identified from neuronal tissue where they play a key role to maintain the vesicular glutamate level. Recently, it has been demonstrated that glutamate signaling is also functional in peripheral neuronal and non-neuronal tissues, and occurs in sites of pituitary, adrenal, pineal glands, bone, GI tract, pancreas,skin, and testis. The glutamate receptors and VGLUTs in digestivesystem have been found in both neuronal and endocrinal cells. The glutamate signaling in the digestive system may have significant relevance to diabetes and GI tract motility disorders. This review will focus on the most recent update of molecular physiology of digestive VGLUTs.

  17. [How to cope with recurrent aphthous stomatitis].

    Science.gov (United States)

    Madrid, C; Jaques, B; Bouferrrache, K; Broome, M

    2010-10-06

    Recurrent aphthous stomatitis (RAS) is the most common oral mucosa ailment. This condition is frequently considered as idiopathic due to the doubts about its etiology, probably related to a minor immunological dysregulation in a context of genetic predisposition. However, ulcers that resemble recurrent aphthous stomatitis in some respects can be found in systemic disorders that must be ruled out for the differential diagnosis of SAR, particularly when they appear after adolescence and/or when associated lesions exist out of the oral cavity. SAR management lies on the elimination of predisposing factors (drugs, oral trauma, food allergies...) and if needed, topical corticosteroids are the first choice regimen. More severe cases may require systemic regimens.

  18. Recurrent aphthous stomatitis: genetic aspects of etiology

    Science.gov (United States)

    Szponar, Elżbieta; Kowalska, Anna

    2013-01-01

    Recurrent aphthous stomatitis (RAS; recurrent aphthous ulcers – RAU; canker sores) is a chronic inflammatory, ulcerative condition of the oral mucosa. Its prevalence in the general population ranges between 5% and 20%, depending on the method and group studied. The etiopathogenesis of the disease is considered to be multifactorial, but remains still not fully understood. In patients with RAS, an enhanced immunologic response occurs to some trigger factors that may include: mechanical injury, stress or bacterial and viral antigens. Higher prevalence of aphthae in relatives may also indicate the genetic background of the condition. The inheritance of some specific gene polymorphisms, especially those encoding proinflammatory cytokines, which play a role in the formation of aphthous ulcer, may predispose family members to RAS. The purpose of this paper was to present the main clinical features of recurrent aphthous stomatitis, epidemiologic data and crucial etiopathogenetic factors with a special emphasis on genetic background of the condition. PMID:24278055

  19. STUDY REGARDING STOMATAL DENSITY IN MAGNOLIA SP.

    Directory of Open Access Journals (Sweden)

    Nicoleta-Valentina GROZA

    2013-06-01

    Full Text Available The purpose of this study is to reveal the structural aspects of the leaf as occurring in the genus Magnolia. The leaves are bifacial and hypostomatic. Secretory oil cells are a constant presence. We have revealed significant dissimilarities in stomatal density and size as occurring in three ornamental species: Magnolia kobus, Magnolia x soulangeana “Soulange-Bodin” (M. denudata x M. liliiflora and Magnolia x “Susan” (M. kobus var. stellata “Rosea” x M. liliiflora “Nigra”. The highest stomatal density was recorded in the diploid species Magnolia kobus. The stomata are significantly elongated in Magnolia x soulangeana “Soul.-Bod.” and wide in Magnolia kobus.

  20. Stochastic Model of Maturation and Vesicular Exchange in Cellular Organelles

    CERN Document Server

    Vagne, Quentin

    2016-01-01

    The dynamical organization of membrane-bound organelles along intracellular transport pathways relies on vesicular exchange between organelles and on biochemical maturation of the organelle content by specific enzymes. The relative importance of each mechanism in controlling organelle dynamics remains controversial, in particular for transport through the Golgi apparatus. Using a stochastic model, we show that full maturation of membrane-bound compartments can be seen as the stochastic escape from a steady-state in which export is dominated by vesicular exchange. We show that full maturation can contribute a significant fraction of the total out-flux for small organelles such as endosomes and Golgi cisternae.

  1. Plant virus infections control stomatal development

    Science.gov (United States)

    Murray, Rose R.; Emblow, Mark S. M.; Hetherington, Alistair M.; Foster, Gary D.

    2016-09-01

    Stomata are important regulators of carbon dioxide uptake and transpirational water loss. They also represent points of vulnerability as bacterial and fungal pathogens utilise this natural opening as an entry portal, and thus have an increasingly complex relationship. Unlike the situation with bacterial and fungal pathogens, we know very little about the role of stomata in viral infection. Here we report findings showing that viral infection influences stomatal development in two susceptible host systems (Nicotiana tabacum with TMV (Tobacco mosaic virus), and Arabidopsis thaliana with TVCV (Turnip vein-clearing virus)), but not in resistant host systems (Nicotiana glutinosa and Chenopodium quinoa with TMV). Virus infected plants had significantly lower stomatal indices in systemic leaves of susceptible systems; N. tabacum 9.8% reduction and A. thaliana 12.3% reduction, but not in the resistant hosts. Stomatal density in systemic leaves was also significantly reduced in virus infected A. thaliana by 19.6% but not in N. tabacum or the resistant systems. In addition, transpiration rate was significantly reduced in TMV infected N. tabacum.

  2. MINYAK GOSOK DAPAT MENGOBATI STOMATITIS APTOSA REKUREN SECARA TOPIKAL

    OpenAIRE

    Ali Yusran; Donald RN

    2016-01-01

    Many studies have been done to get the right material in the treatment of recurrent minor aphthous stomatitis, but the result still unsatisfied. The aim ofthistudy was to know the influence application of rubbing oil against the healing and comportable at recurrent minor aphthous stomatitis. This study was a clinical observasionally and healing was resulted in about four days. The use of topical rubbing oil in the treatment of recurrent minor aphthous stomatitis could...

  3. Vesicular signalling and immune modulation as hedonic fingerprints

    DEFF Research Database (Denmark)

    Bisgaard, Christina F; Bak, Steffen; Christensen, Trine

    2012-01-01

    ) differential gel electrophoresis (DIGE) and tandem mass spectrometry (MS/MS). The majority of the proteins we identified were enzymes involved in different metabolic activities. Additional proteins were functionally classified as vesicular proteins and immune system proteins. Rab GDP dissociation inhibitor...

  4. Formulation and evaluation of a transfersomal vesicular carrier ...

    African Journals Online (AJOL)

    AF1 was investigated. NIPRD-AF1 is a phytomedicine derived from the leaves of an indigenous plant, for use in the treatment of fungal infections. A transfersomal vesicular carrier system of NIPRD-AF1 was formulated and evaluated for topical ...

  5. Effect of vesicular arbuscular mycorrhizal fungus on the ...

    African Journals Online (AJOL)

    The symbiotic association between certain plants and microorganisms plays an important role in soil fertilization, and improves their growth and mineral nutrition. The symbiotic association between vesicular arbuscular mycorrhizal (VAM) fungi and roots provides a significant contribution to plant nutrition and growth.

  6. Renal epithelial cells can release ATP by vesicular fusion

    Directory of Open Access Journals (Sweden)

    Randi G Bjaelde

    2013-09-01

    Full Text Available Renal epithelial cells have the ability to release nucleotides as paracrine factors. In the intercalated cells of the collecting duct, ATP is released by connexin30 (cx30, which is selectively expressed in this cell type. However, ATP is released by virtually all renal epithelia and the aim of the present study was to identify possible alternative nucleotide release pathways in a renal epithelial cell model. We used MDCK (type1 cells to screen for various potential ATP release pathways. In these cells, inhibition of the vesicular H+-ATPases (bafilomycin reduced both the spontaneous and hypotonically (80%-induced nucleotide release. Interference with vesicular fusion using N-ethylamide markedly reduced the spontaneous nucleotide release, as did interference with trafficking from the endoplasmic reticulum to the Golgi apparatus (brefeldin A1 and vesicular transport (nocodazole. These findings were substantiated using a siRNA directed against SNAP-23, which significantly reduced spontaneous ATP release. Inhibition of pannexin and connexins did not affect the spontaneous ATP release in this cell type, which consists of ∼90% principal cells. TIRF-microscopy of either fluorescently-labeled ATP (MANT-ATP or quinacrine-loaded vesicles, revealed that spontaneous release of single vesicles could be promoted by either hypoosmolality (50% or ionomycin. This vesicular release decreased the overall cellular fluorescence by 5.8% and 7.6% respectively. In summary, this study supports the notion that spontaneous and induced ATP release can occur via exocytosis in renal epithelial cells.

  7. The vesicular-arbuscular mycorrhizal symbiosis | Quilambo | African ...

    African Journals Online (AJOL)

    Vesicular-arbuscular mycorrhiza fungi are associated with the majority ot the terrestrial plants. Their function ranges from stress alleviation to bioremediation in soils polluted with heavy metals. However, our knowledge about this symbiosis is still limited. For the semi-arid tropics, where some african countries are located, ...

  8. Influence of vesicular arbuscular mycorrhiza (VAM) and phosphate ...

    African Journals Online (AJOL)

    A field experiment was carried out to find out the effect of biofertilizers, vesicular arbuscular mycorrhiza (VAM), and phosphate solubilising bacteria (PSB) individually and in combination on growth and physiological attributing properties of Marsdenia volubilis plant under nursery conditions. The plant seedlings were ...

  9. Drought induces alterations in the stomatal development program in Populus

    Science.gov (United States)

    Campbell, Malcolm M

    2012-01-01

    Much is known about the physiological control of stomatal aperture as a means by which plants adjust to water availability. By contrast, the role played by the modulation of stomatal development to limit water loss has received much less attention. The control of stomatal development in response to water deprivation in the genus Populus is explored here. Drought induced declines in stomatal conductance as well as an alteration in stomatal development in two genotypes of Populus balsamifera. Leaves that developed under water-deficit conditions had lower stomatal indices than leaves that developed under well-watered conditions. Transcript abundance of genes that could hypothetically underpin drought-responsive changes in stomatal development was examined, in two genotypes, across six time points, under two conditions, well-watered and with water deficit. Populus homologues of STOMAGEN, ERECTA (ER), STOMATA DENSITY AND DISTRIBUTION 1 (SDD1), and FAMA had variable transcript abundance patterns congruent with their role in the modulation of stomatal development in response to drought. Conversely, there was no significant variation in transcript abundance between genotypes or treatments for the Populus homologues of YODA (YDA) and TOO MANY MOUTHS (TMM). The findings highlight the role that could be played by stomatal development during leaf expansion as a longer term means by which to limit water loss from leaves. Moreover, the results point to the key roles played by the regulation of the homologues of STOMAGEN, ER, SDD1, and FAMA in the control of this response in poplar. PMID:22760471

  10. Plant water potential improves prediction of empirical stomatal models.

    Directory of Open Access Journals (Sweden)

    William R L Anderegg

    Full Text Available Climate change is expected to lead to increases in drought frequency and severity, with deleterious effects on many ecosystems. Stomatal responses to changing environmental conditions form the backbone of all ecosystem models, but are based on empirical relationships and are not well-tested during drought conditions. Here, we use a dataset of 34 woody plant species spanning global forest biomes to examine the effect of leaf water potential on stomatal conductance and test the predictive accuracy of three major stomatal models and a recently proposed model. We find that current leaf-level empirical models have consistent biases of over-prediction of stomatal conductance during dry conditions, particularly at low soil water potentials. Furthermore, the recently proposed stomatal conductance model yields increases in predictive capability compared to current models, and with particular improvement during drought conditions. Our results reveal that including stomatal sensitivity to declining water potential and consequent impairment of plant water transport will improve predictions during drought conditions and show that many biomes contain a diversity of plant stomatal strategies that range from risky to conservative stomatal regulation during water stress. Such improvements in stomatal simulation are greatly needed to help unravel and predict the response of ecosystems to future climate extremes.

  11. Stomatal Blue Light Response Is Present in Early Vascular Plants.

    Science.gov (United States)

    Doi, Michio; Kitagawa, Yuki; Shimazaki, Ken-ichiro

    2015-10-01

    Light is a major environmental factor required for stomatal opening. Blue light (BL) induces stomatal opening in higher plants as a signal under the photosynthetic active radiation. The stomatal BL response is not present in the fern species of Polypodiopsida. The acquisition of a stomatal BL response might provide competitive advantages in both the uptake of CO2 and prevention of water loss with the ability to rapidly open and close stomata. We surveyed the stomatal opening in response to strong red light (RL) and weak BL under the RL with gas exchange technique in a diverse selection of plant species from euphyllophytes, including spermatophytes and monilophytes, to lycophytes. We showed the presence of RL-induced stomatal opening in most of these species and found that the BL responses operated in all euphyllophytes except Polypodiopsida. We also confirmed that the stomatal opening in lycophytes, the early vascular plants, is driven by plasma membrane proton-translocating adenosine triphosphatase and K(+) accumulation in guard cells, which is the same mechanism operating in stomata of angiosperms. These results suggest that the early vascular plants respond to both RL and BL and actively regulate stomatal aperture. We also found three plant species that absolutely require BL for both stomatal opening and photosynthetic CO2 fixation, including a gymnosperm, C. revoluta, and the ferns Equisetum hyemale and Psilotum nudum. © 2015 American Society of Plant Biologists. All Rights Reserved.

  12. Analysis of Stomatal Patterning in Selected Mutants of MAPK Pathways

    KAUST Repository

    Felemban, Abrar

    2016-05-01

    Stomata are cellular valves in plants that play an essential role in the regulation of gas exchange and are distributed in the epidermis of aerial organs. In Arabidopsis thaliana, stomatal production and development are coordinated by the mitogen-activated protein kinase (MAPK) signalling pathway, which modulates a variety of other processes, including cell proliferation, regulation of cytokinesis, programed cell death, and response to abiotic and biotic stress. The environment also plays a role in stomatal development, by influencing the frequency at which stomata develop in leaves. This thesis presents an analysis of stomatal development in Arabidopsis mutants in two MAPK pathways: MEKK1-MKK1/MKK2-MPK4, and MAP3K17/18-MKK3. Obtained results demonstrate the effect of stress conditions on stomatal development and specify the involvement of analysed MAPK in stomatal patterning. First, both analysed pathways modulate stomatal patterning in Arabidopsis cotyledons. Second, plant growth-promoting bacteria tested enhance stomatal density and affect guard cell morphology. Third, the sucrose or mannitol treatment increases defects in stomatal patterning. Finally, salt stress or high temperature can suppress stomatal defects in mutants of the MEKK1-MKK1/MKK2-MPK4 pathway.

  13. Foot & Mouth Disease & Ulcerative/Vesicular Rule-outs: Challenges Encountered in Recent Outbreaks

    Energy Technology Data Exchange (ETDEWEB)

    Hullinger, P

    2008-01-28

    development and subsequent rupturing of vesicles at the coronary band and in the oral cavity. Vesicles and ulcerations can also occur on the mammary gland. Recovery in adult animals usually occurs in 8-15 days. Clinical signs for most serotypes are less dramatic in sheep and goats. Swine can develop very severe coronary band lesions and high mortality in piglets has been observed. One of the challenges of diagnosing FMD is that it may be clinically similar to several other vesicular or ulcerative diseases. FMD is clinically indistinguishable from Vesicular stomatitis, Swine vesicular disease and Vesicular exanthema of swine. It may also resemble Bovine viral diarrhea, Mucosal disease, Infectious bovine rhinotracheitis, Bluetongue, Bovine papular stomatitis, Bovine mammillitis and Rinderpest.

  14. Total antioxidant status and oxidative stress in recurrent aphthous stomatitis.

    Science.gov (United States)

    Tugrul, Selahattin; Koçyiğit, Abdurrahim; Doğan, Remzi; Eren, Sabri Baki; Senturk, Erol; Ozturan, Orhan; Ozar, Omer Faruk

    2016-03-01

    Recurrent aphthous stomatitis is an idiopathic, chronic, recurrent inflammatory disease of the oral mucosa. It is thought that oxidative stress caused by systemic inflammation plays a basic role in the etiopathogenesis of recurrent aphthous stomatitis. The aim of this study is to review oxidative status and DNA damage in recurrent aphthous stomatitis. The study included 42 patients with an active recurrent aphthous stomatitis lesion and 39 healthy volunteers with similar demographic characteristics. DNA damage was analyzed using alkaline single cell gel electrophoresis (comet assay). Plasma levels of total antioxidant status and total oxidative status were determined by using an automated measurement method. Oxidative stress index was calculated as total oxidative status/total antioxidant status and × 100. The total oxidative status and oxidative stress index values were significantly higher in the recurrent aphthous stomatitis group compared to the control group, while total antioxidant status values were significantly lower. In the recurrent aphthous stomatitis group, DNA damage was observed to be significantly higher than the control group. In correlation analysis, significant correlation was found between DNA damage and the oxidative stress index and total oxidative status values in the recurrent aphthous stomatitis group. This is the first report in the literature that demonstrates association of recurrent aphthous stomatitis with increased oxidative status. © 2015 The International Society of Dermatology.

  15. Stomatal characteristics of Eucalyptus grandis clonal hybrids in ...

    African Journals Online (AJOL)

    The aim was to investigate the degree to which stomatal conductance (gs) and stomatal density differ between the clonal hybrids across seasons and in response to water stress. Plants from one E. grandis x E. camaldulensis (GC) and two E. grandis x E. urophylla (GU1 and GU2) clones were grown for 18 months in 80 l ...

  16. Dynamic changes of stomatal characteristics during the flower, fruit ...

    African Journals Online (AJOL)

    The results showed that there were stomata on the median region of exocarps, adaxial and abaxial epidermis of the petals and leaf midribs. The petal and fruit epidermal cells were polygonal in shape, while leaf epidermal cells were strip. The leaf stomatal index and stomatal density were the largest in the surfaces ...

  17. Quantitative trait loci mapping for stomatal traits in interspecific ...

    Indian Academy of Sciences (India)

    Dr.YASODHA

    Cartographer v 2.5 (Wang et al. 2007) to identify QTLs for stomatal density, stomatal area and pore length in adaxial and abaxial leaf surfaces adopting backcross model. The LOD threshold was determined by permutation analysis with 1000 repetitions. The size of the analysis window was maintained at 10 cM with a walk ...

  18. Urban Legends Series: Recurrent Aphthous Stomatitis

    Science.gov (United States)

    Baccaglini, Lorena; Lalla, Rajesh V.; Bruce, Alison J.; Sartori-Valinotti, Julio C.; Latortue, Marie C.; Carrozzo, Marco; Rogers, Roy S.

    2011-01-01

    Recurrent aphthous stomatitis (RAS) is the most common idiopathic intraoral ulcerative disease in the USA. Aphthae typically occur in apparently healthy individuals, although an association with certain systemic diseases has been reported. Despite the unclear etiopathogenesis, new drug trials are continuously conducted in an attempt to reduce pain and dysfunction. We investigated four controversial topics: (1) Is complex aphthosis a mild form of Behçet’s disease (BD)? (2) Is periodic fever, aphthous stomatitis, pharyngitis and adenitis (PFAPA) syndrome a distinct medical entity? (3) Is RAS associated with other systemic diseases (e.g., celiac disease and B12 deficiency)? (4) Are there any new RAS treatments? Results from extensive literature searches, including a systematic review of RAS trials, suggested that: (1) Complex aphthosis is not a mild form of BD in North America or Western Europe; (2) Diagnostic criteria for PFAPA have low specificity and the characteristics of the oral ulcers warrant further studies; (3) Oral ulcers may be associated with celiac disease; however, these ulcers may not be RAS; RAS is rarely associated with B12 deficiency; nevertheless, B12 treatment may be beneficial, via mechanisms that warrant further study; (4) Thirty-three controlled trials published in the past 6 years reported some effectiveness, though potential for bias was high. PMID:21812866

  19. [Vesicular and pronuclear glycoproteins in the pathogenesis of cholesterol lithiasis].

    Science.gov (United States)

    Jirsa, M; Smíd, F; Marecek, Z

    1998-01-26

    Several biliary proteins have been known to accelerate fusion of cholesterol rich phospholipid vesicles. Some of them are present in vesicular membrane, localisation of other proteins is unknown. Biliary glycoprotein has not been studied in consequence with pathogenesis of cholesterol lithiasis. Low molecular extravesicular proteins were separated from vesicles by gel filtration on a 1200mm column of Sephacryl S-300 HR. Immunoglobulins IgM, IgA, haptoglobin, biliary glycoprotein I (BGP I) and nonspecific crossreactive antigen were eluted along with vesicles. Albumin and alpha 1-acid glycoprotein were eluted later and must be extravesicular. Fact that BGP I (85 kDa membrane glycoprotein) eluted along with vesicles and not in albumin fraction suggests that it might be bound in vesicular membrane. As a known adhesion molecule it could thus play an important role in pathogenesis of cholesterol cholelithiasis.

  20. Expression of Vesicular Nucleotide Transporter in Rat Odontoblasts

    OpenAIRE

    Ikeda, Erina; Goto, Tetsuya; Gunjigake, Kaori; Kuroishi, Kayoko; Ueda, Masae; Kataoka, Shinji; Toyono, Takashi; Nakatomi, Mitsushiro; Seta, Yuji; Kitamura, Chiaki; Nishihara, Tatsuji; Kawamoto, Tatsuo

    2016-01-01

    Several theories have been proposed regarding pain transmission mechanisms in tooth. However, the exact signaling mechanism from odontoblasts to pulp nerves remains to be clarified. Recently, ATP-associated pain transmission has been reported, but it is unclear whether ATP is involved in tooth pain transmission. In the present study, we focused on the vesicular nucleotide transporter (VNUT), a transporter of ATP into vesicles, and examined whether VNUT was involved in ATP release from odontob...

  1. Influenza infection modulates vesicular trafficking and induces Golgi complex disruption.

    Science.gov (United States)

    Yadav, Vibha; Panganiban, Antonito T; Honer Zu Bentrup, Kerstin; Voss, Thomas G

    2016-12-01

    Influenza A virus (IFV) replicates its genome in the nucleus of infected cells and uses the cellular protein transport system for genome trafficking from the nucleus to the plasma membrane. However, many details of the mechanism of this process, and its relationship to subsequent cytoplasmic virus trafficking, have not been elucidated. We examined the effect of nuclear transport inhibitors Leptomycin B (LB), 5,6 dichloro-1-β-d-ribofuranosyl-benzimidazole (DRB), the vesicular transport inhibitor Brefeldin A (BFA), the caspase inhibitor ZWEHD, and microtubule inhibitor Nocodazole (NOC) on virus replication and intracellular trafficking of viral nucleoprotein (NP) from the nucleus to the ER and Golgi. Also, we carried out complementary studies to determine the effect of IFV on intracellular membranes. Inhibition of the CRM1 and TAP-P15 nuclear transport pathways by DRB and LB blocked completely the export of virus. Inhibition of vesicular trafficking by BFA, NOC, and ZWEHD also affected influenza infection. Interestingly, IFV infection induced fragmentation of the Golgi complex resulting in diffuse distribution of large and small vesicles throughout the cytoplasm. Live-cell microscopy revealed expansion of Golgi localization signals indicating progressive dispersion of Golgi positive structures, resulting in the disassembly of the Golgi ribbon structure. Other vesicular components (Rab1b, ARF1 and GBF1) were also found to be required for IFV infection. Furthermore, the exact step at which IFV infection disrupts vesicle trafficking was identified as the ER-Golgi intermediate compartment. These findings suggest that IFV NP is trafficked from the nucleus via the CRM1 and TAP pathways. IFV modulates vesicular trafficking inducing disruption of the Golgi complex. These studies provide insight on the ways in which IFV affects intracellular trafficking of different host proteins and will facilitate identification of useful pharmaceutical targets to abrogate virus

  2. Management of Recurrent Aphthous Stomatitis in Children.

    Science.gov (United States)

    Montgomery-Cranny, Jodie A; Wallace, Ann; Rogers, Helen J; Hughes, Sophie C; Hegarty, Anne M; Zaitoun, Halla

    2015-01-01

    Recurrent oral ulceration is common and may present in childhood. Causes of recurrent oral ulceration are numerous and there may be an association with underlying systemic disease. Recurrent aphthous stomatitis (RAS) is the most common underlying diagnosis in children. The discomfort of oral ulcers can impact negatively on quality of life of a child, interfering with eating, speaking and may result in missed school days. The role of the general dental practitioner is to identify patients who can be treated with simple measures in primary dental care and those who require assessment and treatment in secondary care. Management may include topical agents for symptomatic relief, topical corticosteroids and, in severe recalcitrant cases, systemic agents may be necessary.

  3. Haematological parameters and recurrent aphthous stomatitis.

    Science.gov (United States)

    Khan, Nabiha Farasat; Saeed, Mohammad; Chaudhary, Saima; Khan, Nabiha Farasat

    2013-02-01

    To find out the relationship between recurrent aphthous stomatitis (RAS) with deficiencies of haemoglobin, haematocrit, serum vitamin B12, serum Ferritin and red blood cells (RBC) Folate level. An analytical cross-sectional study. Department of Oral Health Sciences, Shaikh Zayed Federal Postgraduate Medical Complex, Lahore, from February to July 2008. Sixty consecutive subjects with active RAS were taken as the aphthous group; 60 age and gender matched subjects without RAS were as the Non-Aphthous group. Five milliliter blood was taken from both groups to evaluate the levels of serum B12, and RBC Folate through radio immuno assay and serum ferritin with enzyme linked immuno-sorbent assay tests. Complete blood count was carried out to determine the level of haemoglobin and haematocrit in both groups. Proportion of subjects with lower values was compared using 2 text of proportions with significance at p aphthous group.

  4. Recurrent aphthous stomatitis and Helicobacter pylori.

    Science.gov (United States)

    Gomes, Carolina-Cavaliéri; Gomez, Ricardo-Santiago; Zina, Lívia-Guimarães; Amaral, Fabrício-Rezende

    2016-03-01

    Recurrent aphthous stomatitis (RAS) is a recurrent painful ulcerative disorder that commonly affects the oral mucosa. Local and systemic factors such as trauma, food sensitivity, nutritional deficiencies, systemic conditions, immunological disorders and genetic polymorphisms are associated with the development of the disease. Helicobacter pylori (H. pylori) is a gram-negative, microaerophile bacteria, that colonizes the gastric mucosa and it was previously suggested to be involved in RAS development. In the present paper we reviewed all previous studies that investigated the association between RAS and H. pylori. A search in Pubmed (MEDLINE) databases was made of articles published up until July 2015 using the following keywords: Helicobacter Pylori or H. pylori and RAS or Recurrent aphthous stomatitis. Fifteen experimental studies that addressed the relationship between infection with H. pylori and the presence of RAS and three reviews, including a systematic review and a meta-analysis were included in this review. The studies reviewed used different methods to assess this relationship, including PCR, nested PCR, culture, ELISA and urea breath test. A large variation in the number of patients included in each study, as well as inclusion criteria and laboratorial methods was observed. H. pylori can be detected in the oral mucosa or ulcerated lesion of some patients with RAS. The quality of the all studies included in this review was assessed using levels of evidence based on the University of Oxford's Center for Evidence Based Medicine Criteria. Although the eradication of the infection may affect the clinical course of the oral lesions by undetermined mechanisms, RAS ulcers are not associated with the presence of the bacteria in the oral cavity and there is no evidence that H. pylori infection drives RAS development.

  5. Origins and Evolution of Stomatal Development1[OPEN

    Science.gov (United States)

    2017-01-01

    The fossil record suggests stomata-like pores were present on the surfaces of land plants over 400 million years ago. Whether stomata arose once or whether they arose independently across newly evolving land plant lineages has long been a matter of debate. In Arabidopsis, a genetic toolbox has been identified that tightly controls stomatal development and patterning. This includes the basic helix-loop-helix (bHLH) transcription factors SPEECHLESS (SPCH), MUTE, FAMA, and ICE/SCREAMs (SCRMs), which promote stomatal formation. These factors are regulated via a signaling cascade, which includes mobile EPIDERMAL PATTERNING FACTOR (EPF) peptides to enforce stomatal spacing. Mosses and hornworts, the most ancient extant lineages to possess stomata, possess orthologs of these Arabidopsis (Arabidopsis thaliana) stomatal toolbox genes, and manipulation in the model bryophyte Physcomitrella patens has shown that the bHLH and EPF components are also required for moss stomatal development and patterning. This supports an ancient and tightly conserved genetic origin of stomata. Here, we review recent discoveries and, by interrogating newly available plant genomes, we advance the story of stomatal development and patterning across land plant evolution. Furthermore, we identify potential orthologs of the key toolbox genes in a hornwort, further supporting a single ancient genetic origin of stomata in the ancestor to all stomatous land plants. PMID:28356502

  6. Vesicular exanthema of swine virus: isolation and serotyping of field samples.

    OpenAIRE

    Edwards, J F; Yedloutschnig, R J; Dardiri, A H; Callis, J. J.

    1987-01-01

    Virus isolation was attempted from 262 field samples of vesicular material collected during the outbreaks of vesicular exanthema of swine in the U.S.A. from 1952-54. Using primary swine kidney culture, viral cytopathogenic agents were isolated from 76.3% of the samples. However, an overall recovery rate of 82.1% was obtained after samples negative in tissue culture were inoculated intradermally in susceptible swine. All vesicular exanthema of swine virus isolates were identified as serotype B...

  7. Role of Sucrose in Emerging Mechanisms of Stomatal Aperture Regulation.

    Energy Technology Data Exchange (ETDEWEB)

    Outlaw, W. H.

    2000-09-15

    Focused on the second of 2 hypotheses that were proposed for testing that transpiration rate determines the extent to which suc accumulates in the GC wall providing a mechanism for regulating stomatal aperture size.

  8. Endobronchial tuberculosis presented as multiple endobronchial vesicular lesions

    Directory of Open Access Journals (Sweden)

    Farah Idrees

    2015-01-01

    Full Text Available Endobronchial tuberculosis (EBTB is a tuberculous infection of the tracheobronchial tree with microbiological and histopathological evidence, with or without parenchymal involvement. EBTB commonly presents as acute or insidious onset cough, wheeze, low grade fever, and constitutional symptoms. In elderly patients, other differentials like malignancy and pneumonia may lead to misdiagnosis. Hence, bronchoscopy is essential for confirmation of EBTB. Here we report a rare presentation of EBTB in a 65 year old patient who presented with 3 months history of fever and cough and have multiple endobronchial vesicular lesions on bronchoscopy.

  9. Vesicular-Arbuscular Mycorrhiza in Field-Grown Crops

    DEFF Research Database (Denmark)

    Jakobsen, Iver

    1986-01-01

    The importance of vesicular-arbuscular mycorrhiza (VAM) and P fertilizer for P nutrition and dry matter production in field peas (Pisum sativum L.) was studied in moderately P-deficient soil. Half of the experimental plots were fumigated to reduce the level of VAM infection. Shoots and 0 to 30 cm...... in fumigated plots, although both it and P uptake were increased by adding P fertilizer. The possible reasons for this discrepancy are discussed. A supplementary survey on infection development at five other field sites showed that peas are extensively colonized by VAM fungi, even in soils where a standard...

  10. The acclimation of Tilia cordata stomatal opening in response to light, and stomatal anatomy to vegetational shade and its components.

    Science.gov (United States)

    Aasamaa, Krõõt; Aphalo, Pedro José

    2017-02-01

    Stomatal anatomical traits and rapid responses to several components of visible light were measured in Tilia cordata Mill. seedlings grown in an open, fully sunlit field (C-set), or under different kinds of shade. The main questions were: (i) stomatal responses to which visible light spectrum regions are modified by growth-environment shade and (ii) which separate component of vegetational shade is most effective in eliciting the acclimation effects of the full vegetational shade. We found that stomatal opening in response to red or green light did not differ between the plants grown in the different environments. Stomatal response to blue light was increased (in comparison with that of C-set) in the leaves grown in full vegetational shade (IABW-set), in attenuated UVAB irradiance (AB-set) or in decreased light intensity (neutral shade) plus attenuated UVAB irradiance (IAB-set). In all sets, the addition of green light-two or four times stronger-into induction light barely changed the rate of the blue-light-stimulated stomatal opening. In the AB-set, stomatal response to blue light equalled the strong IABW-set response. In attenuated UVB-grown leaves, stomatal response fell midway between IABW- and C-set results. Blue light response by neutral shade-grown leaves did not differ from that of the C-set, and the response by the IAB-set did not differ from that of the AB-set. Stomatal size was not modified by growth environments. Stomatal density and index were remarkably decreased only in the IABW- and IAB-sets. It was concluded that differences in white light responses between T. cordata leaves grown in different light environments are caused only by their different blue light response. Differences in stomatal sensitivity are not dependent on altered stomatal anatomy. Attenuated UVAB irradiance is the most efficient component of vegetational shade in stimulating acclimation of stomata, whereas decreased light intensity plays a minor role. © The Author 2016. Published

  11. Helicobacter pylori DNA in recurrent aphthous stomatitis.

    Science.gov (United States)

    Victória, Júnia Maria Netto; Kalapothakis, Evanguedes; Silva, Jeane de Fátima Correia; Gomez, Ricardo Santiago

    2003-04-01

    Considering not only the fact that recurrent aphthous stomatitis (RAS) and stomach ulcers are immunologically mediated ulcers associated with Helicobacter pylori, but also the recent evidence that anaemia can be associated with both diseases, and the discovery of H. pylori in the oral mucosa led us to hypothesize that this bacteria may be related to RAS pathogenesis. Thirty-six consecutive subjects affected by minor and major forms of RAS and 48 healthy volunteers were included in the present study. The nested polymerase chain reaction (PCR) technique was used to detect the presence of H. pylori in the oral lesion, the normal contralateral mucosa of patients affected by RAS and the oral mucosa of control subjects. The chi2- and Fisher's tests were used for statistical analysis. No association between RAS lesions and H. pylori was observed. However, 14 out of 36 (38.9%) of the patients with RAS were found to show the presence of H. pylori DNA in the lesion and/or contralateral mucosa. Sixteen out of 48 (33.3%) of the patients without RAS (control subjects) were positive (P > 0.05). The present study does not give support to the assumption that H. pylori could be involved in RAS development.

  12. Oxidative stress and recurrent aphthous stomatitis.

    Science.gov (United States)

    Bagan, Jose; Saez, Guillermo; Tormos, Carmen; Gavalda, Carmen; Sanchis, Jose M; Bagan, Leticia; Scully, Crispian

    2014-11-01

    This study analyzed the oxidative stress status in patients with recurrent aphthous stomatitis (RAS) in the presence and absence of active ulceration. Oxidative stress was analyzed in peripheral mononuclear cells of 28 RAS patients with active ulceration and 29 controls. A further blood sample was collected from nine subjects randomly selected from the 28 RAS cases, during the period in which the patients did not have active oral ulceration. The reduced glutathione (GSH), malondialdehyde (MDA), and oxidized glutathione (GSSG) levels were measured in these samples. The mean MDA and GSSG levels were significantly higher in patients with active RAS than in the controls, while GSH was lower in the RAS group (p < 0.01). There was a nonsignificant tendency toward higher MDA and GSSG levels in patients with major RAS compared with minor RAS. On comparing the serum findings in the nine RAS patients in the presence and absence of lesions, the presence of ulceration was associated with even higher MDA and GSSG levels and lower GSH concentrations (p < 0.05) CONCLUSIONS: Oxidative stress was detected in our RAS patients.

  13. Psychological stress and recurrent aphthous stomatitis

    Directory of Open Access Journals (Sweden)

    Camila de Barros Gallo

    2009-01-01

    Full Text Available INTRODUCTION AND OBJECTIVES: Recurrent aphthous stomatitis (RAS is the most common type of ulcerative disease of the oral mucosa. Despite its worldwide occurrence and the extensive amount of research that has been devoted to the subject, the etiology of RAS remains unclear. Nevertheless, several hereditary, nutritional, infectious and psychological factors have been associated with RAS. The aim of this case-control study was to assess the influence of psychological stress on the manifestation of RAS. METHOD: Fifty patients were enrolled in the trial. Twenty-five RAS patients constituted the study group and another 25 non-RAS patients who were similarly matched for sex, age and socioeconomic status constituted the control group. Each patient was evaluated in terms of the four domains of stress (emotional, physical, social and cognitive using an internationally validated questionnaire, which was comprised of 59 items and measured the frequency and intensity of stress symptoms. The RAS group was interviewed during an active RAS episode. Completed questionnaires were submitted to proper analytical software and interpreted by an expert psychologist. RESULTS: There was a higher level of psychological stress among RAS group patients when compared to the control group (P < 0.05. CONCLUSION: Psychological stress may play a role in the manifestation of RAS; it may serve as a trigger or a modifying factor rather than being a cause of the disease.

  14. Recurrent aphthous stomatitis: a case report

    Directory of Open Access Journals (Sweden)

    Xiomara Serpa-Romero

    2016-07-01

    Full Text Available Recurrent aphthosus stomatitis is an alteration of the oral mucosa in some cases associated with depression of the immune system that affects the tissue response at the level of the epithelium, triggering repetitive clinical picture of small and medium ulcers (3-5 mm which necrotic presented erythematous background and lasting no more than 15 days. The picture becomes recurrent, symptomatic, compromising the health of the patient who consults again with the same characteristics in oral cavity. The literature associates the process with hormonal changes, trauma, prolonged intake of medications, and stress. A case of female patient 53, who attends the service of dentistry to present multiple oral thrush that hard to swallow, drooling and feverish marked presents in Santa Marta, at the Center for Implantology and Oral Rehabilitation. According to the interrogation and clinical examination it is associated with a reactive inflammatory process caused by the intake of drugs to treat infectious or viral process, which is given the presumptive diagnosis of erythema drug. Any medication intake was suspended and additional tests are ordered antinuclear antibodies

  15. Stomatal malfunctioning under low VPD conditions: induced by alterations in stomatal morphology and leaf anatomy or in the ABA signaling?

    Science.gov (United States)

    Aliniaeifard, Sasan; Malcolm Matamoros, Priscila; van Meeteren, Uulke

    2014-12-01

    Exposing plants to low VPD reduces leaf capacity to maintain adequate water status thereafter. To find the impact of VPD on functioning of stomata, stomatal morphology and leaf anatomy, fava bean plants were grown at low (L, 0.23 kPa) or moderate (M, 1.17 kPa) VPDs and some plants that developed their leaves at moderate VPD were then transferred for 4 days to low VPD (M→L). Part of the M→L-plants were sprayed with ABA (abscisic acid) during exposure to L. L-plants showed bigger stomata, larger pore area, thinner leaves and less spongy cells compared with M-plants. Stomatal morphology (except aperture) and leaf anatomy of the M→L-plants were almost similar to the M-plants, while their transpiration rate and stomatal conductance were identical to that of L-plants. The stomatal response to ABA was lost in L-plants, but also after 1-day exposure of M-plants to low VPD. The level of foliar ABA sharply decreased within 1-day exposure to L, while the level of ABA-GE (ABA-glucose ester) was not affected. Spraying ABA during the exposure to L prevented loss of stomatal closing response thereafter. The effect of low VPD was largely depending on exposure time: the stomatal responsiveness to ABA was lost after 1-day exposure to low VPD, while the responsiveness to desiccation was gradually lost during 4-day exposure to low VPD. Leaf anatomical and stomatal morphological alterations due to low VPD were not the main cause of loss of stomatal closure response to closing stimuli. © 2014 Scandinavian Plant Physiology Society.

  16. Distribution of vesicular glutamate transporters in the human brain

    Directory of Open Access Journals (Sweden)

    Erika eVigneault

    2015-03-01

    Full Text Available Glutamate is the major excitatory transmitter in the brain. Vesicular glutamate transporters (VGLUT1-3 are responsible for uploading glutamate into synaptic vesicles. VGLUT1 and VGLUT2 are considered as specific markers of canonical glutamatergic neurons, while VGLUT3 is found in neurons previously shown to use other neurotransmitters than glutamate. Although there exists a rich literature on the localization of these glutamatergic markers in the rodent brain, little is currently known about the distribution of VGLUT1-3 in the human brain. In the present study, using subtype specific probes and antisera, we examined the localization of the three vesicular glutamate transporters in the human brain by in situ hybridization, immunoautoradiography and immunohistochemistry. We found that the VGLUT1 transcript was highly expressed in the cerebral cortex, hippocampus and cerebellum, whereas VGLUT2 mRNA was mainly found in the thalamus and brainstem. VGLUT3 mRNA was localized in scarce neurons within the cerebral cortex, hippocampus, striatum and raphe nuclei. Following immunoautoradiographic labeling, intense VGLUT1- and VGLUT2-immunoreactivities were observed in all regions investigated (cerebral cortex, hippocampus, caudate-putamen, cerebellum, thalamus, amygdala, substantia nigra, raphe while VGLUT3 was absent from the thalamus and cerebellum. This extensive mapping of VGLUT1-3 in human brain reveals distributions that correspond for the most part to those previously described in rodent brains.

  17. Comparación de la técnica seroneutralización con la inmunoenzimática competitiva en la detección de anticuerpos contra el virus de la estomatitis vesicular (serotipos New Jersey e Indiana) en equinos de Costa Rica

    OpenAIRE

    von Koeckritz-Blickwede, Maren; Dolz Wiedner, Gaby; Vinicio Herrero, Marco

    2015-01-01

    A competitive enzyme-linked immunosorbent assay (c-ELISA) was compared to the serum neutralization test (SNT) for the detection of antibodies against vesicular stomatitis virus, New Jersey (VSV-NJ) and Indiana (VSV-IN) serotypes, using 214 equine serum samples from Costa Rica. In the c-ELISA for VSV-NJ 109 (50.93%) equine sera reacted positive, whereas in the SNT 138 (64.48%) serum samples reacted positive to VSV-NJ. In the c-ELISA for VSV-IN 26 (12.15%) sera reacted positive, whereas in the ...

  18. Vesicular disease in 9-week-old pigs experimentally infected with Senecavirus A

    Science.gov (United States)

    Introduction: Senecavirus A (SVA), a picornavirus, has been infrequently associated with cases of idiopathic vesicular disease (IVD) in pigs in the US and Canada since 1988. In 2014 and 2015 there was surge of IVD cases in Brazil and US, respectively. SVA was identified in serum, vesicular fluid, an...

  19. H-aggregation of azobenzene-substituted amphiphiles in vesicular membranes

    NARCIS (Netherlands)

    Kuiper, JM; Engberts, JBFN

    2004-01-01

    Photochemical switching has been studied of double-tailed phosphate amphiphiles containing azobenzene units in both tails in aqueous vesicular dispersions and in mixed vesicular systems with 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC). Since the ease of switching depends on the strength of the

  20. Chronic gingivitis and aphthous stomatitis relationship hypothesis: A neuroimmunobiological approach

    Directory of Open Access Journals (Sweden)

    Chiquita Prahasanti

    2009-03-01

    Full Text Available Background: Traumatic injuries to the oral mucosa in fixed orthodontic patients are common, especially in the first week of bracket placement, and occasionally lead to the development of aphthous stomatitis or ulcers. Nevertheless, these lesions are selflimiting. Purpose: The objective of this study is to reveal the connection between chronic gingivitis and aphthous stomatitis which is still unclear. Case: A patient with a persistent lesion for more than six months. Case Management: RAS was treated with scaling procedure, the gingival inflammation was healed. However, in this case report, despite the appropriate management procedures had been done, the lesion still worsen and became more painful. Moreover, the symptoms did not heal for more than two weeks. Actually, they had been undergone orthodontic treatment more than six months and rarely suffered from aphthous stomatitis. Coincidentally, at that time they also suffered from chronic gingivitis. It was interesting that after scaling procedures, the ulcer subsides in two days. Conclusion: Recently, the neuroimmunobiological researches which involved neurotransmitters and cytokines on cell-nerve signaling, and heat shock proteins in gingivitis and stomatitis are in progress. Nevertheless, they were done separately, thus do not explain the interrelationship. This proposed new concept which based on an integrated neuroimmunobiological approach could explain the benefit of periodontal treatment, especially scaling procedures, for avoiding prolonged painful episodes and unnecessary medications in aphthous stomatitis. However, for widely acceptance of the chronic gingivitis and aphthous stomatitis relationship, further clinical and laboratory study should be done. Regarding to the relatively fast healing after scaling procedures in this case report; it was concluded that the connection between chronic gingivitis and aphthous stomatitis is possible.

  1. Multivitamin therapy for recurrent aphthous stomatitis

    Science.gov (United States)

    Lalla, Rajesh V.; Choquette, Linda E.; Feinn, Richard S.; Zawistowski, Harriet; Latortue, Marie C.; Kelly, Edward T.; Baccaglini, Lorena

    2013-01-01

    Background Recurrent aphthous stomatitis (RAS) is a painful condition of unknown etiology, affecting more than 2.5 billion people worldwide. Vitamin deficiencies have been implicated as a possible cause. Methods The authors conducted a single-center, randomized, parallel-arm, double-masked, placebo-controlled study to examine the effect of daily multivitamin supplementation on the number and duration of RAS episodes. The authors randomly assigned 160 adults who had a validated history of at least three episodes of idiopathic minor RAS within the previous 12 months to one of two groups: the first group (n = 83) received a once-daily multivitamin containing 100 percent of the U.S. reference daily intake (RDI) of essential vitamins, and the second group (n = 77) received once-daily placebo for up to 365 days. Results The results showed no significant difference in the mean number of new RAS episodes between the multivitamin (4.19 episodes) and placebo (4.60 episodes) arms during the study period (P = .69). The mean duration of new RAS episodes also was similar for the multivitamin (8.66 days) and placebo (8.99 days) arms (P = .60). Furthermore, the authors found no differences between the two arms with regard to mouth pain, normalcy of diet or compliance with the study medication regimen. Conclusion Daily multivitamin supplementation, with the RDI of essential vitamins, did not result in a reduction in the number or duration of RAS episodes. Clinical Implications Clinicians should not recommend multi-vitamin supplementation routinely as prophylaxis for RAS. PMID:22467697

  2. A putative vesicular transporter expressed in Drosophila mushroom bodies that mediates sexual behavior may define a novel neurotransmitter system

    OpenAIRE

    Brooks, Elizabeth S.; Greer, Christina L.; Romero-Calderón, Rafael; Serway, Christine N.; Grygoruk, Anna; Haimovitz, Jasmine M.; Bac T. Nguyen; Najibi, Rod; Tabone, Christopher J; de Belle, J. Steven; Krantz, David E.

    2011-01-01

    Storage and release of classical and amino acid neurotransmitters requires vesicular transporters. Some neurons lack known vesicular transporters, suggesting additional neurotransmitter systems remain unidentified. Insect mushroom bodies (MBs) are critical for several behaviors, including learning, but the neurotransmitters released by the intrinsic Kenyon cells (KCs) remain unknown. Likewise, KCs do not express a known vesicular transporter. We report the identification of a novel Drosophila...

  3. Evolutionary Conservation of ABA Signaling for Stomatal Closure1[OPEN

    Science.gov (United States)

    Huang, Yuqing; Dai, Fei; Franks, Peter J.; Nevo, Eviatar; Soltis, Douglas E.; Soltis, Pamela S.; Xue, Dawei; Zhang, Guoping; Pogson, Barry J.

    2017-01-01

    Abscisic acid (ABA)-driven stomatal regulation reportedly evolved after the divergence of ferns, during the early evolution of seed plants approximately 360 million years ago. This hypothesis is based on the observation that the stomata of certain fern species are unresponsive to ABA, but exhibit passive hydraulic control. However, ABA-induced stomatal closure was detected in some mosses and lycophytes. Here, we observed that a number of ABA signaling and membrane transporter protein families diversified over the evolutionary history of land plants. The aquatic ferns Azolla filiculoides and Salvinia cucullata have representatives of 23 families of proteins orthologous to those of Arabidopsis (Arabidopsis thaliana) and all other land plant species studied. Phylogenetic analysis of the key ABA signaling proteins indicates an evolutionarily conserved stomatal response to ABA. Moreover, comparative transcriptomic analysis has identified a suite of ABA-responsive genes that differentially expressed in a terrestrial fern species, Polystichum proliferum. These genes encode proteins associated with ABA biosynthesis, transport, reception, transcription, signaling, and ion and sugar transport, which fit the general ABA signaling pathway constructed from Arabidopsis and Hordeum vulgare. The retention of these key ABA-responsive genes could have had a profound effect on the adaptation of ferns to dry conditions. Furthermore, stomatal assays have shown the primary evidence for ABA-induced closure of stomata in two terrestrial fern species P. proliferum and Nephrolepis exaltata. In summary, we report, to our knowledge, new molecular and physiological evidence for the presence of active stomatal control in ferns. PMID:28232585

  4. A new role for human dyskerin in vesicular trafficking.

    Science.gov (United States)

    Di Maio, Nunzia; Vicidomini, Rosario; Angrisani, Alberto; Belli, Valentina; Furia, Maria; Turano, Mimmo

    2017-10-01

    Dyskerin is an essential, conserved, multifunctional protein found in the nucleolus, whose loss of function causes the rare genetic diseases X-linked dyskeratosis congenita and Hoyeraal-Hreidarsson syndrome. To further investigate the wide range of dyskerin's biological roles, we set up stable cell lines able to trigger inducible protein knockdown and allow a detailed analysis of the cascade of events occurring within a short time frame. We report that dyskerin depletion quickly induces cytoskeleton remodeling and significant alterations in endocytic Ras-related protein Rab-5A/Rab11 trafficking. These effects arise in different cell lines well before the onset of telomere shortening, which is widely considered the main cause of dyskerin-related diseases. Given that vesicular trafficking affects many homeostatic and differentiative processes, these findings add novel insights into the molecular mechanisms underlining the pleiotropic manifestation of the dyskerin loss-of-function phenotype.

  5. [CORRELATION MATRIX OF CHARACTERISTICS OF CHRONIC RECURRENT APHTHOUS STOMATITIS].

    Science.gov (United States)

    Koridze, Kh; Aladashvili, L; Taboridze, I

    2015-09-01

    The purpose of the present work is to study the correlation between the risk factors of chronic recurrent aphthous stomatitis. The research was conducted on 62 patients between ages of 40 and 70 years at Tbilisi Hospital for Veterans of War. The analysis was carried out by Spearman's Rank Correlation method using the statistical package SPSS 11.5. We investigated: harmful habits, professional factors, background and accompanying illnesses, pathology of teeth, focal infection, emotional stress, genetic factors. Correlation matrix between the significant risk factors of chronic recurrent aphthous stomatitis is defined. Multiple correlations have the following factors: industrial dust, focal infections, emotional stress, anemia. Correlation diagram of etiological factors of chronic recurrent aphthous stomatitis is helpful for providing professional and expert services.

  6. Incidence of Helicobacter Pylori in oral aphthous stomatitis

    Directory of Open Access Journals (Sweden)

    Surender Sharma

    2016-01-01

    Full Text Available Background/Aims: The aim of this study was to determine probable HP infection in oral aphthous samples by RUT in patients with recurrent aphthous stomatitis. Materials and Methods: This in vivo cross-sectional study was approved by the Ethics Committee of Swami Devi Dayal Dental College and Hospital according to the ethical standards. A total of 30 patients with minor recurrent aphthous stomatitis and 20 healthy control groups were included in the study. Results: Out of 30 patients with minor recurrent aphthous stomatitis, including 17 male and 13 female patients, with mean age of 47 and 38 years respectively, 21 patients (70% were RUT (positive. Out of 20 healthy control groups, 12 males and 8 females, 2 (10% were RUT (positive [Table 1]. Conclusion: HP may play a role in the etiology of RAS; also it is likely that RUT may be rapid and reliable for investigation of HP in RAS lesions.

  7. Treatment of radiation- and chemotherapy-induced stomatitis

    Energy Technology Data Exchange (ETDEWEB)

    Carnel, S.B.; Blakeslee, D.B.; Oswald, S.G.; Barnes, M. (Fitzsimons Army Medical Center, Aurora, CO (USA))

    1990-04-01

    Severe stomatitis is a common problem encountered during either radiation therapy or chemotherapy. Most therapeutic regimens are empirical, with no scientific basis. The purpose of this study is to determine the efficacy of various topical solutions in the treatment of radiation- or chemotherapy-induced stomatitis. Eighteen patients were entered into a prospective double-blinded study to test several topical solutions: (1) viscous lidocaine with 1% cocaine; (2) dyclonine hydrochloride 1.0% (Dyclone); (3) kaolin-pectin solution, diphenhydramine plus saline (KBS); and (4) a placebo solution. Degree of pain relief, duration of relief, side effects, and palatability were evaluated. The results showed that Dyclone provided the most pain relief. Dyclone and viscous lidocaine with 1% cocaine provided the longest pain relief, which averaged 50 minutes This study provides objective data and defines useful guidelines for treatment of stomatitis.

  8. Are vesicular neurotransmitter transporters potential treatment targets for temporal lobe epilepsy?

    Directory of Open Access Journals (Sweden)

    Joeri eVan Liefferinge

    2013-08-01

    Full Text Available The vesicular neurotransmitter transporters (VNTs are small proteins responsible for packing synaptic vesicles with neurotransmitters thereby determining the amount of neurotransmitter released per vesicle through fusion in both neurons and glial cells. Each transporter subtype was classically seen as a specific neuronal marker of the respective nerve cells containing that particular neurotransmitter or structurally related neurotransmitters. More recently, however, it has become apparent that common neurotransmitters can also act as co-transmitters, adding complexity to neurotransmitter release and suggesting intriguing roles for VNTs therein. We will first describe the current knowledge on vesicular glutamate transporters (VGLUT1/2/3, the vesicular excitatory amino acid transporter (VEAT, the vesicular nucleotide transporter (VNUT, vesicular monoamine transporters (VMAT1/2, the vesicular acetylcholine transporter (VAChT and the vesicular γ-aminobutyric acid (GABA transporter (VGAT in the brain. We will focus on evidence regarding transgenic mice with disruptions in VNTs in different models of seizures and epilepsy. We will also describe the known alterations and reorganizations in the expression levels of these VNTs in rodent models for temporal lobe epilepsy (TLE and in human tissue resected for epilepsy surgery. Finally, we will discuss perspectives on opportunities and challenges for VNTs as targets for possible future epilepsy therapies.

  9. Vesicular nucleotide transporter (VNUT): appearance of an actress on the stage of purinergic signaling.

    Science.gov (United States)

    Moriyama, Yoshinori; Hiasa, Miki; Sakamoto, Shohei; Omote, Hiroshi; Nomura, Masatoshi

    2017-06-14

    Vesicular storage of ATP is one of the processes initiating purinergic chemical transmission. Although an active transport mechanism was postulated to be involved in the processes, a transporter(s) responsible for the vesicular storage of ATP remained unidentified for some time. In 2008, SLC17A9, the last identified member of the solute carrier 17 type I inorganic phosphate transporter family, was found to encode the vesicular nucleotide transporter (VNUT) that is responsible for the vesicular storage of ATP. VNUT transports various nucleotides in a membrane potential-dependent fashion and is expressed in the various ATP-secreting cells. Mice with knockout of the VNUT gene lose vesicular storage and release of ATP from neurons and neuroendocrine cells, resulting in blockage of the initiation of purinergic chemical transmission. Thus, VNUT plays an essential role in the vesicular storage and release of ATP. The VNUT knockout mice exhibit resistance for neuropathic pain and a therapeutic effect against diabetes by way of increased insulin sensitivity. Thus, VNUT inhibitors and suppression of VNUT gene expression may be used for therapeutic purposes through suppression of purinergic chemical transmission. This review summarizes the studies to date on VNUT and discusses what we have learned about the relevance of vesicular ATP release as a potential drug target.

  10. Stomatal Opening Involves Polar, Not Radial, Stiffening Of Guard Cells.

    Science.gov (United States)

    Carter, Ross; Woolfenden, Hugh; Baillie, Alice; Amsbury, Sam; Carroll, Sarah; Healicon, Eleanor; Sovatzoglou, Spyros; Braybrook, Sioban; Gray, Julie E; Hobbs, Jamie; Morris, Richard J; Fleming, Andrew J

    2017-10-09

    It has long been accepted that differential radial thickening of guard cells plays an important role in the turgor-driven shape changes required for stomatal pore opening to occur [1-4]. This textbook description derives from an original interpretation of structure rather than measurement of mechanical properties. Here we show, using atomic force microscopy, that although mature guard cells display a radial gradient of stiffness, this is not present in immature guard cells, yet young stomata show a normal opening response. Finite element modeling supports the experimental observation that radial stiffening plays a very limited role in stomatal opening. In addition, our analysis reveals an unexpected stiffening of the polar regions of the stomata complexes, both in Arabidopsis and other plants, suggesting a widespread occurrence. Combined experimental data (analysis of guard cell wall epitopes and treatment of tissue with cell wall digesting enzymes, coupled with bioassay of guard cell function) plus modeling lead us to propose that polar stiffening reflects a mechanical, pectin-based pinning down of the guard cell ends, which restricts increase of stomatal complex length during opening. This is predicted to lead to an improved response sensitivity of stomatal aperture movement with respect to change of turgor pressure. Our results provide new insight into the mechanics of stomatal function, both negating an established view of the importance of radial thickening and providing evidence for a significant role for polar stiffening. Improved stomatal performance via altered cell-wall-mediated mechanics is likely to be of evolutionary and agronomic significance. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  11. Modeling stomatal conductance and ozone uptake of Fagus crenata grown under different nitrogen loads.

    Science.gov (United States)

    Azuchi, Fumika; Kinose, Yoshiyuki; Matsumura, Tomoe; Kanomata, Tomoaki; Uehara, Yui; Kobayashi, Ayumi; Yamaguchi, Masahiro; Izuta, Takeshi

    2014-01-01

    A multiplicative stomatal conductance model was constructed to estimate stomatal O3 uptake of Fagus crenata exposed to O3 under different N loads to the soil. Our stomatal conductance model included environmental functions such as the stomatal responses of F. crenata to diurnal changes, chronic O3 stress (AOT0), acute O3 stress (O3 concentration), and nitrogen load to soil. The model could explain 62% of the variability in stomatal conductance. We suggest therefore that stomatal closure induced by O3 and N load-induced soil acidification must be taken into account in developing a stomatal conductance model for estimating stomatal O3 uptake for future risk assessment of O3 impact on Japanese forest tree species such as F. crenata. Copyright © 2013 Elsevier Ltd. All rights reserved.

  12. Measurement of Stomatal Aperture by Digital Image Processing

    OpenAIRE

    Kenji, Omasa; Morio, Onoe; Division of Engineering The National Institute for Environmental Studies; Institute of Industrial Science, University of Tokyo

    1984-01-01

    We developed a new digital image processing technique for exactly measuring the degree of stomatal opening, that is, the ratio of the width to the maximum length of a stomatal pore, and the pore area. We applied this technique to evaluate responses to SO_2 of neighboring stomata in a small region of an intact attached leaf, with the following results: 1) The pore region could be exactly extracted even when the original digital image was of poor quality. The standard errors in the evaluation o...

  13. Optimal Stomatal Behaviour Around the World: Synthesis of a Global Stomatal Conductance Database and Scaling from Leaf to Ecosystem

    Science.gov (United States)

    Lin, Y. S.; Medlyn, B. E.; Duursma, R.; Prentice, I. C.; Wang, H.

    2014-12-01

    Stomatal conductance (gs) is a key land surface attribute as it links transpiration, the dominant component of global land evapotranspiration and a key element of the global water cycle, and photosynthesis, the driving force of the global carbon cycle. Despite the pivotal role of gs in predictions of global water and carbon cycles, a global scale database and an associated globally applicable model of gs that allow predictions of stomatal behaviour are lacking. We present a unique database of globally distributed gs obtained in the field for a wide range of plant functional types (PFTs) and biomes. We employed a model of optimal stomatal conductance to assess differences in stomatal behaviour, and estimated the model slope coefficient, g1, which is directly related to the marginal carbon cost of water, for each dataset. We found that g1 varies considerably among PFTs, with evergreen savanna trees having the largest g1 (least conservative water use), followed by C3 grasses and crops, angiosperm trees, gymnosperm trees, and C4 grasses. Amongst angiosperm trees, species with higher wood density had a higher marginal carbon cost of water, as predicted by the theory underpinning the optimal stomatal model. There was an interactive effect between temperature and moisture availability on g1: for wet environments, g1 was largest in high temperature environments, indicated by high mean annual temperature during the period when temperature above 0oC (Tm), but it did not vary with Tm across dry environments. We examine whether these differences in leaf-scale behaviour are reflected in ecosystem-scale differences in water-use efficiency. These findings provide a robust theoretical framework for understanding and predicting the behaviour of stomatal conductance across biomes and across PFTs that can be applied to regional, continental and global-scale modelling of productivity and ecohydrological processes in a future changing climate.

  14. Spatial heterogeneity in stomatal features during leaf elongation: an analysis using Rosa hybrida

    NARCIS (Netherlands)

    Fanourakis, D.; Heuvelink, E.; Carvalho, S.M.P.

    2015-01-01

    Within-leaf heterogeneity in stomatal traits poses a key uncertainty in determining a representative value for the whole leaf. Accounting for this heterogeneity, we studied stomatal initiation on expanding leaves and estimated stomatal conductance (gs) of mature leaves. The entire lamina was

  15. Jasmonate-mediated stomatal closure under elevated CO2 revealed by time-resolved metabolomics

    Science.gov (United States)

    Foliar stomatal movements are critical for regulating plant water status and gas exchange. Elevated carbon dioxide (CO2) concentrations are known to induce stomatal closure. However, current knowledge on CO2 signal transduction in stomatal guard cells is limited. Here we report the metabolomic respo...

  16. Mammalian target of rapamycin inhibitor-associated stomatitis

    NARCIS (Netherlands)

    Boers-Doets, Christine B.; Raber-Durlacher, Judith E.; Treister, Nathaniel S.; Epstein, Joel B.; Arends, Anniek B. P.; Wiersma, Diede R.; Lalla, Rajesh V.; Logan, Richard M.; van Erp, Nielka P.; Gelderblom, Hans

    2013-01-01

    With the recent introduction of inhibitors of mammalian target of rapamycin (mTOR) in oncology, distinct cutaneous and oral adverse events have been identified. In fact, stomatitis and rash are documented as the most frequent and potentially dose-limiting side effects. Clinically, mTOR

  17. Detection of salivary interleukin-2 in recurrent aphthous stomatitis.

    Science.gov (United States)

    Kalpana, R; Thubashini, M; Sundharam, B Sivapatha

    2014-01-01

    The present study was undertaken to estimate and compare salivary interleukin-2 (IL-2) levels in patients with recurrent aphthous stomatitis, among healthy controls and their variation with age and sex. Saliva was collected from 60 patients within the age range of 16-60 years which included 30 patients (17 Females and 13 Males) with recurrent aphthous stomatitis and healthy control group consisted of 30 participants (18 Females and 12 Males). IL-2 estimation was done in both the groups using enzyme linked immunosorbent assay (ELISA). Statistical analysis of the data was done using Independent 't' test. The results showed increased salivary IL-2 levels in patients with recurrent aphthous stomatitis compared to the healthy controls. The IL-2 levels were also increased in patients with the age group of 16-30 years compared to other age groups. Similar increase of IL-2 was also seen in female patients. Age related and sex related alterations of IL-2 in recurrent aphthous stomatitis patients were observed.

  18. mechanisms of drought resistance in grain ii:.stomatal regulation ...

    African Journals Online (AJOL)

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    whereby photosythesis was reduced by 75% in common bean but only by 20% in chickpea. Significant ... induced stomatal closure, thereby a decrease in the ... 7 or 15 days at soil water content of 40% (-0.15 MPa. SWP). ... chamber were kept constant with a light intensity ... determination of highway distances on maps. At.

  19. Influence of climate variables on Cyperus papyrus stomatal ...

    African Journals Online (AJOL)

    Cyperus papyrus forms highly productive wetlands in tropical Africa, but the environmental control of transpirational water loss in wetlands is poorly understood. The influence of climate variables on papyrus stomatal conductance in dry and wet seasons of the year was investigated in a wetland in Kampala, Uganda, ...

  20. Effects of Moisture and Mycorrhiza on Stomatal Conductance and ...

    African Journals Online (AJOL)

    acer

    Both the ecto and endo mycorrhizae significantly (P<0.05)affected the Stomatal Conductance and Xylem Pressure Potential of the plant even under stressed conditions. The use of mycorrhiza is thus recommended as a strategy for efficient water utilization and water conservation. KEYWORDS: Faidherbia albida, mycorrhiza, ...

  1. Linking stomatal sensitivity and whole-tree hydraulic architecture

    Science.gov (United States)

    Katherine A. McCulloh; David R. Woodruff

    2012-01-01

    Despite the complexity of the relationship between stomatal sensitivity, water loss and vulnerability to embolism, the goal of teasing apart the subtleties is a necessary one. As Litvak et al. (2012) mention, determining transpiration patterns based on vulnerability to embolism would be much easier than the lengthy and potentially expensive processes involved in sap...

  2. Improvement of herpetic stomatitis therapy in patients with chronic tonsillitis

    Directory of Open Access Journals (Sweden)

    Lepilin А.V.

    2011-12-01

    Full Text Available The research goal is to determine the clinical and pathogenetic efficacy of Cycloferon liniment in the combined therapy in patients with herpetic stomatitis accompanied by chronic tonsillitis. Materials and methods: Medical examination and treatment of 60 patients have been carried out. The marker of endogenous intoxication, infectious severity and immunity has been investigated. Results. It has been established that use of Cycloferon liniment in the combined therapy in patients with herpetic stomatitis accompanied by chronic tonsillitis has allowed to decrease infectious severity in par-odontal recess and evidence of local inflammation, to normalize immunity indices and reduce the level of endogenous intoxication that has been liable for acceleration of recuperation processes and lowering of frequency of stomatitis recurrences. Conclusion. The clinical efficacy of Cycloferon liniment in the therapy in patients with herpetic stomatitis accompanied by chronic tonsillitis conditioned by the decreasing of activity of local inflammatory process according to the reducing of level pro-inflammatory cytokines, infectious burden of the mouth cavity, endogenous intoxication

  3. Comparison of different stomatal conductance algorithms for ozone flux modelling

    NARCIS (Netherlands)

    Büker, P.; Emberson, L.D.; Ashmore, M.R.; Gerosa, G.; Jacobs, C.M.J.; Massman, W.J.; Müller, J.; Nikolov, N.; Novak, K.; Oksanen, E.; Torre, de la D.; Tuovinen, J.P.

    2007-01-01

    A multiplicative and a semi-mechanistic, BWB-type [Ball, J.T., Woodrow, I.E., Berry, J.A., 1987. A model predicting stomatal conductance and its contribution to the control of photosynthesis under different environmental conditions. In: Biggens, J. (Ed.), Progress in Photosynthesis Research, vol.

  4. Comparison of different stomatal conductance algorithms for ozone flux modelling

    Science.gov (United States)

    P. Buker; L.D. Emberson; M. R. Ashmore; H. M. Cambridge; C. M. Jacobs; W. J. Massman; J. Muller; N. Nikolov; K. Novak; E. Oksanen; M. Schaub; D. de la Torre

    2007-01-01

    A multiplicative and a semi-mechanistic, BWB-type [Ball, J.T., Woodrow, I.E., Berry, J.A., 1987. A model predicting stomatal conductance and its contribution to the control of photosynthesis under different environmental conditions. In: Biggens, J. (Ed.), Progress in Photosynthesis Research, vol. IV. Martinus Nijhoff, Dordrecht, pp. 221-224.] algorithm for calculating...

  5. Evolutionary Conservation of ABA Signaling for Stomatal Closure.

    Science.gov (United States)

    Cai, Shengguan; Chen, Guang; Wang, Yuanyuan; Huang, Yuqing; Marchant, D Blaine; Wang, Yizhou; Yang, Qian; Dai, Fei; Hills, Adrian; Franks, Peter J; Nevo, Eviatar; Soltis, Douglas E; Soltis, Pamela S; Sessa, Emily; Wolf, Paul G; Xue, Dawei; Zhang, Guoping; Pogson, Barry J; Blatt, Michael R; Chen, Zhong-Hua

    2017-06-01

    Abscisic acid (ABA)-driven stomatal regulation reportedly evolved after the divergence of ferns, during the early evolution of seed plants approximately 360 million years ago. This hypothesis is based on the observation that the stomata of certain fern species are unresponsive to ABA, but exhibit passive hydraulic control. However, ABA-induced stomatal closure was detected in some mosses and lycophytes. Here, we observed that a number of ABA signaling and membrane transporter protein families diversified over the evolutionary history of land plants. The aquatic ferns Azolla filiculoides and Salvinia cucullata have representatives of 23 families of proteins orthologous to those of Arabidopsis (Arabidopsis thaliana) and all other land plant species studied. Phylogenetic analysis of the key ABA signaling proteins indicates an evolutionarily conserved stomatal response to ABA. Moreover, comparative transcriptomic analysis has identified a suite of ABA-responsive genes that differentially expressed in a terrestrial fern species, Polystichum proliferum These genes encode proteins associated with ABA biosynthesis, transport, reception, transcription, signaling, and ion and sugar transport, which fit the general ABA signaling pathway constructed from Arabidopsis and Hordeum vulgare The retention of these key ABA-responsive genes could have had a profound effect on the adaptation of ferns to dry conditions. Furthermore, stomatal assays have shown the primary evidence for ABA-induced closure of stomata in two terrestrial fern species Pproliferum and Nephrolepis exaltata In summary, we report, to our knowledge, new molecular and physiological evidence for the presence of active stomatal control in ferns. © 2017 American Society of Plant Biologists. All Rights Reserved.

  6. Expression of Vesicular Nucleotide Transporter in Rat Odontoblasts.

    Science.gov (United States)

    Ikeda, Erina; Goto, Tetsuya; Gunjigake, Kaori; Kuroishi, Kayoko; Ueda, Masae; Kataoka, Shinji; Toyono, Takashi; Nakatomi, Mitsushiro; Seta, Yuji; Kitamura, Chiaki; Nishihara, Tatsuji; Kawamoto, Tatsuo

    2016-02-27

    Several theories have been proposed regarding pain transmission mechanisms in tooth. However, the exact signaling mechanism from odontoblasts to pulp nerves remains to be clarified. Recently, ATP-associated pain transmission has been reported, but it is unclear whether ATP is involved in tooth pain transmission. In the present study, we focused on the vesicular nucleotide transporter (VNUT), a transporter of ATP into vesicles, and examined whether VNUT was involved in ATP release from odontoblasts. We examined the expression of VNUT in rat pulp by RT-PCR and immunostaining. ATP release from cultured odontoblast-like cells with heat stimulation was evaluated using ATP luciferase methods. VNUT was expressed in pulp tissue, and the distribution of VNUT-immunopositive vesicles was confirmed in odontoblasts. In odontoblasts, some VNUT-immunopositive vesicles were colocalized with membrane fusion proteins. Additionally P2X3, an ATP receptor, immunopositive axons were distributed between odontoblasts. The ATP release by thermal stimulation from odontoblast-like cells was inhibited by the addition of siRNA for VNUT. These findings suggest that cytosolic ATP is transported by VNUT and that the ATP in the vesicles is then released from odontoblasts to ATP receptors on axons. ATP vesicle transport in odontoblasts seems to be a key mechanism for signal transduction from odontoblasts to axons in the pulp.

  7. Transferosomes - A vesicular transdermal delivery system for enhanced drug permeation

    Directory of Open Access Journals (Sweden)

    Reshmy Rajan

    2011-01-01

    Full Text Available Transdermal administration of drugs is generally limited by the barrier function of the skin. Vesicular systems are one of the most controversial methods for transdermal delivery of active substances. The interest in designing transdermal delivery systems was relaunched after the discovery of elastic vesicles like transferosomes, ethosomes, cubosomes, phytosomes, etc. This paper presents the composition, mechanisms of penetration, manufacturing and characterization methods of transferosomes as transdermal delivery systems of active substances. For a drug to be absorbed and distributed into organs and tissues and eliminated from the body, it must pass through one or more biological membranes/barriers at various locations. Such a movement of drug across the membrane is called as drug transport. For the drugs to be delivered to the body, they should cross the membranous barrier. The concept of these delivery systems was designed in an attempt to concentrate the drug in the tissues of interest, while reducing the amount of drug in the remaining tissues. Hence, surrounding tissues are not affected by the drug. In addition, loss of drug does not happen due to localization of drug, leading to get maximum efficacy of the medication. Therefore, the phospholipid based carrier systems are of considerable interest in this era.

  8. Transferosomes - A vesicular transdermal delivery system for enhanced drug permeation

    Science.gov (United States)

    Rajan, Reshmy; Jose, Shoma; Mukund, V. P. Biju; Vasudevan, Deepa T.

    2011-01-01

    Transdermal administration of drugs is generally limited by the barrier function of the skin. Vesicular systems are one of the most controversial methods for transdermal delivery of active substances. The interest in designing transdermal delivery systems was relaunched after the discovery of elastic vesicles like transferosomes, ethosomes, cubosomes, phytosomes, etc. This paper presents the composition, mechanisms of penetration, manufacturing and characterization methods of transferosomes as transdermal delivery systems of active substances. For a drug to be absorbed and distributed into organs and tissues and eliminated from the body, it must pass through one or more biological membranes/barriers at various locations. Such a movement of drug across the membrane is called as drug transport. For the drugs to be delivered to the body, they should cross the membranous barrier. The concept of these delivery systems was designed in an attempt to concentrate the drug in the tissues of interest, while reducing the amount of drug in the remaining tissues. Hence, surrounding tissues are not affected by the drug. In addition, loss of drug does not happen due to localization of drug, leading to get maximum efficacy of the medication. Therefore, the phospholipid based carrier systems are of considerable interest in this era. PMID:22171309

  9. [Safety of laparoscopic cholecystectomy in complicated vesicular disease].

    Science.gov (United States)

    Pérez-Morales, Alfonso; Roesch-Dietlen, Federico; Díaz-Blanco, Fernando; Martínez-Fernández, Silvia

    2005-01-01

    We assessed the experience and safety of cholecystectomy through laparoscopic approach in patients with complicated biliary gallbladder disease who were attended at the Hospital Español, Veracruz, Mexico, during a 10-year period. A prospective, longitudinal, comparative study with a control group was designed. We studied a group of patients with complicated cholelithiasis disease (Group I) and compared them with patients without complicated diseases. We analyzed the following variables: age, gender, risk factors, associated trans-operative pathology and accidents, surgical time, rate of conversion to open procedure, length of hospital stay, complications and evolution. During the 10-year period, 733 cholecystectomies were performed, 245 (33.42%) to treat complicated cholelithiasis and 488 (66.58%) uncomplicated. There were no differences regarding gender, age, risk factors, hospital stay, complications, morbidity and mortality, and iatrogenic lesions of the biliary tree, postoperative morbidity and mortality. In all patients of Group I (245), we found complications of cholelithiasis (acute cholecystitis, choledochal gallstones, vesicular adherences, and cholecystocolonic fistula). Surgical time was longer in Group I and surgical accidents made the surgical procedure more difficult. The overall results established that laparoscopic cholecystectomy in our institution is a safe procedure in patients with complicated gallbladder disease and can be performed by experienced surgeons.

  10. The BIG protein distinguishes the process of CO2-induced stomatal closure from the inhibition of stomatal opening by CO2.

    Science.gov (United States)

    He, Jingjing; Zhang, Ruo-Xi; Peng, Kai; Tagliavia, Cecilia; Li, Siwen; Xue, Shaowu; Liu, Amy; Hu, Honghong; Zhang, Jingbo; Hubbard, Katharine E; Held, Katrin; McAinsh, Martin R; Gray, Julie E; Kudla, Jörg; Schroeder, Julian I; Liang, Yun-Kuan; Hetherington, Alistair M

    2018-04-01

    We conducted an infrared thermal imaging-based genetic screen to identify Arabidopsis mutants displaying aberrant stomatal behavior in response to elevated concentrations of CO 2 . This approach resulted in the isolation of a novel allele of the Arabidopsis BIG locus (At3g02260) that we have called CO 2 insensitive 1 (cis1). BIG mutants are compromised in elevated CO 2 -induced stomatal closure and bicarbonate activation of S-type anion channel currents. In contrast with the wild-type, they fail to exhibit reductions in stomatal density and index when grown in elevated CO 2 . However, like the wild-type, BIG mutants display inhibition of stomatal opening when exposed to elevated CO 2 . BIG mutants also display wild-type stomatal aperture responses to the closure-inducing stimulus abscisic acid (ABA). Our results indicate that BIG is a signaling component involved in the elevated CO 2 -mediated control of stomatal development. In the control of stomatal aperture by CO 2 , BIG is only required in elevated CO 2 -induced closure and not in the inhibition of stomatal opening by this environmental signal. These data show that, at the molecular level, the CO 2 -mediated inhibition of opening and promotion of stomatal closure signaling pathways are separable and BIG represents a distinguishing element in these two CO 2 -mediated responses. © 2018 The Authors. New Phytologist © 2018 New Phytologist Trust.

  11. Rescue at nonpermissive temperature of complementation group II temperature-sensitive mutants of vesicular stomatitis virus by uv-irradiated VSV

    Energy Technology Data Exchange (ETDEWEB)

    Deutsch, V.; Brun, G.

    1978-06-01

    Rescue is group-characteristic. The helper virus can be either the wt strain or a mutant belonging to any group of ts mutants except group II. With regard to genotype, the rescue progeny virus is temperature-sensitive and belongs to group II, and its ts II parent (ts O52(II)) can be characterized. As for phenotype, the in vitro thermal stability of rescue virions is intermediate between that of parental ts O52(II) and uv-irradiated wt virus, suggesting incorporation of some wt protein II molecules in the rescue virions. Different slopes (zero or different from zero) were seen in dose-effect curves representing rescue obtained by structural protein molecules, suggesting that protein II structural role could be distinguished from its functional role(s) by uv sensitivity. Differences in efficiency of the rescue of ts O52(II) by ts I mutants irradiated with low uv fluence may reflect their different transcribing capabilities at 39.6/sup 0/. The results are discussed taking into account the fact that the phenotype of group II mutants is characterized by an unstable nucleocapsid.

  12. Inhibition of EHMT2 induces a robust antiviral response against Foot-and-Mouth Disease and Vesicular Stomatitis Virus infections in bovine cells

    Science.gov (United States)

    The genetic regulatory network controlling the innate immune system is well understood in many species. However, the role of the epigenetic mechanisms underlying the expression of immunoregulatory genes is less clear, especially in livestock species. Histone H3 lysine 9 dimethylation (H3K9me2) is an...

  13. Stomatal malfunctioning under low VPD conditions: induced by alterations in stomatal morphology and leaf anatomy or in the ABA signaling?

    NARCIS (Netherlands)

    Ali Niaei Fard, S.; Malcolm Matamoros, P.; Meeteren, van U.

    2014-01-01

    Exposing plants to low VPD reduces leaf capacity to maintain adequate water status thereafter. To find the impact of VPD on functioning of stomata, stomatal morphology and leaf anatomy, fava bean plants were grown at low (L, 0.23 kPa) or moderate (M, 1.17 kPa) VPDs and some plants that developed

  14. Ozone-induced stomatal sluggishness changes carbon and water balance of temperate deciduous forests.

    Science.gov (United States)

    Hoshika, Yasutomo; Katata, Genki; Deushi, Makoto; Watanabe, Makoto; Koike, Takayoshi; Paoletti, Elena

    2015-05-06

    Tropospheric ozone concentrations have increased by 60-100% in the Northern Hemisphere since the 19(th) century. The phytotoxic nature of ozone can impair forest productivity. In addition, ozone affects stomatal functions, by both favoring stomatal closure and impairing stomatal control. Ozone-induced stomatal sluggishness, i.e., a delay in stomatal responses to fluctuating stimuli, has the potential to change the carbon and water balance of forests. This effect has to be included in models for ozone risk assessment. Here we examine the effects of ozone-induced stomatal sluggishness on carbon assimilation and transpiration of temperate deciduous forests in the Northern Hemisphere in 2006-2009 by combining a detailed multi-layer land surface model and a global atmospheric chemistry model. An analysis of results by ozone FACE (Free-Air Controlled Exposure) experiments suggested that ozone-induced stomatal sluggishness can be incorporated into modelling based on a simple parameter (gmin, minimum stomatal conductance) which is used in the coupled photosynthesis-stomatal model. Our simulation showed that ozone can decrease water use efficiency, i.e., the ratio of net CO2 assimilation to transpiration, of temperate deciduous forests up to 20% when ozone-induced stomatal sluggishness is considered, and up to only 5% when the stomatal sluggishness is neglected.

  15. Vesicular Contact Reaction May Progress into Erythema Multiforme.

    Science.gov (United States)

    Czarnecka-Operacz, Magdalena; Jenerowicz, Dorota; Szulczyńska-Gabor, Joanna; Teresiak-Mikołajczak, Ewa; Szyfter-Harris, Joanna; Bowszyc-Dmochowska, Monika

    2016-12-01

    Dear Editor, Erythema multiforme is considered an acute skin condition, characterized by a self-limiting and sometimes recurrent course. It is regarded as a type IV hypersensitivity reaction associated with certain infections, medications, and other various triggers. Allergic contact dermatitis is in turn a delayed type of induced allergy as a result of cutaneous contact with a specific allergen to which the patient develops specific sensitivity. This type of cutaneous reaction is associated with inflammation manifesting with erythema, edema, and vesicles. A 27-year old female patient presented with a 3-day history of erythematous and vesicular lesions which developed 24 hours after cesarean section. Initially the lesions were localized in the area of surgery (mainly the abdomen and upper thighs) and on the next day progressed to the buttocks and lumbar area. The patient was referred to the Outpatient Clinic and was treated with antihistamines, but her dermatological state deteriorated rapidly. At the day of admission to the Department of Dermatology, numerous erythematous and vesicular lesions were present on the skin of the abdomen, thighs, and back (Figure 1, a), but the skin of the neck, chest, and extremities was also covered with erythematous and edematous patches. On the second day of hospitalization, we observed the evolution of lesions localized within the chest and extremities into an erythema multiforme-like targetoid eruption (Figure 1, b). Initially the patient was treated with intravenous injections of dexamethasone and ceftriaxone and orally with second-generation antihistamines (in four-fold doses), followed by intravenous metyloprednisolone pulse-therapy (total dose of 3 g). As the new vesicobullous lesions started to appear on the face and arms, we introduced cyclosporine A orally 400 mg daily. We could then observe gradual remission, but on the seventh day of hospitalization the patient developed a massive labial herpes simplex infection and had

  16. [Treatment of chronic aphthous stomatitis combined with duodenal ulcer].

    Science.gov (United States)

    Dudchenko, M A; Skrypnikova, T P; Dudchenko, M A

    2014-01-01

    It is currently proved ulcerous stomatitis and duodenal ulcer to have common pathogenetic infectious link (the most studied agent being Helicobacter pylori) by concominant decrease of local and general immunity with hyperoxidation events. Eighty patients (44 female and 36 male aged 15-60) with chronic aphthous stomatitis (AS) combined with duodenal ulcer were included in the study and divided in two equal groups according to treatment received (control group of 40 patients was treated according to conventional scheme, while in 40 patients a new formulation Vipromak was added to treatment protocol). The symptoms of AS tend to resolve faster in vipromak group thus proving its efficiency in treatment of AS and duodenal ulcer.

  17. Remission of severe aphthous stomatitis of celiac disease with etanercept

    Science.gov (United States)

    2013-01-01

    Celiac disease is a common autoimmune disease triggered by gluten-containing foods (wheat, barley and rye) in genetically predisposed individuals. We present a patient with celiac disease complicated by severe aphthous stomatitis resulting in impairing swallowing, chewing and speaking. This led to weight loss, psychosocial problems as well as inability to perform her work. A variety of topical and systemic medications used resulted in either no improvement or only partial alleviation of the patient’s symptoms. After informed consent, etanercept was initiated and resulted in complete remission of aphthous stomatitis, decrease in arthralgia and fatigue and considerable improvement in her quality of life. The use of newer biological agents for selected and severe manifestations of celiac disease may lead to improved morbidity in these patients, but more studies are needed to determine long-term efficacy as well as safety of these drugs in the mucosal and/or systemic complications of this disease. PMID:24365222

  18. Recurrent Aphthous Stomatitis: Diagnosis and Management in Primary Care

    Directory of Open Access Journals (Sweden)

    Jillian Hudson

    2014-11-01

    Full Text Available Recurrent aphthous stomatitis (canker sores is a very common oral condition that remains incompletely understood. Presentation has been well-classified into minor, major or herpetiform subcategories based on clinical features, but exact etiology is unknown. Because etiology is unclear, treatments are primarily empiric and aimed at symptom reduction rather than prevention or cure. However, there are several methods, both topical and systemic, that can be easily and affordably utilized in the primary care setting.

  19. Recurrent Aphthous Stomatitis: Diagnosis and Management in Primary Care

    OpenAIRE

    Jillian Hudson

    2014-01-01

    Recurrent aphthous stomatitis (canker sores) is a very common oral condition that remains incompletely understood. Presentation has been well-classified into minor, major or herpetiform subcategories based on clinical features, but exact etiology is unknown. Because etiology is unclear, treatments are primarily empiric and aimed at symptom reduction rather than prevention or cure. However, there are several methods, both topical and systemic, that can be easily and affordably utilized in the ...

  20. Increasing stomatal conductance in response to rising atmospheric CO2.

    Science.gov (United States)

    Purcell, C; Batke, S P; Yiotis, C; Caballero, R; Soh, W K; Murray, M; McElwain, J C

    2018-01-31

    Studies have indicated that plant stomatal conductance (gs) decreases in response to elevated atmospheric CO2, a phenomenon of significance for the global hydrological cycle. However, gs increases across certain CO2 ranges have been predicted by optimization models. The aim of this work was to demonstrate that under certain environmental conditions, gs can increase in response to elevated CO2. Using (1) an extensive, up-to-date synthesis of gs responses in free air CO2 enrichment (FACE)experiments, (2) in situ measurements across four biomes showing dynamic gs responses to a CO2 rise of ~50 ppm (characterizing the change in this greenhouse gas over the past three decades) and (3) a photosynthesis-stomatal conductance model, it is demonstrated that gs can in some cases increase in response to increasing atmospheric CO2. Field observations are corroborated by an extensive synthesis of gs responses in FACE experiments showing that 11.8 % of gs responses under experimentally elevated CO2 are positive. They are further supported by a strong data-model fit (r2 = 0.607) using a stomatal optimization model applied to the field gs dataset. A parameter space identified in the Farquhar-Ball-Berry photosynthesis-stomatal conductance model confirms field observations of increasing gs under elevated CO2 in hot dry conditions. Contrary to the general assumption, positive gs responses to elevated CO2, although relatively rare, are a feature of woody taxa adapted to warm, low-humidity conditions, and this response is also demonstrated in global simulations using the Community Land Model (CLM4). The results contradict the over-simplistic notion that global vegetation always responds with decreasing gs to elevated CO2, a finding that has important implications for predicting future vegetation feedbacks on the hydrological cycle at the regional level.

  1. Ozone treatment of recurrent aphthous stomatitis: a double blinded study

    OpenAIRE

    AL-Omiri, Mahmoud K.; Mohannad Alhijawi; AlZarea, Bader K.; Ra’ed S. Abul Hassan; Edward Lynch

    2016-01-01

    This study aimed to evaluate the use of ozone to treat recurrent aphthous stomatitis (RAS). Consecutive sixty-nine participants with RAS were recruited into this non-randomized double blind, controlled cohort observational study (test group). A control group of 69 RAS patients who matched test group with age and gender was recruited. RAS lesions in test group were exposed to ozone in air for 60?seconds while controls received only air. Ulcer size and pain were recorded for each participant at...

  2. Oral exfoliative cytology in female reverse smokers having stomatitis nicotina.

    Science.gov (United States)

    Reddy, C R; Sarma, P R; Kameswari, V R

    1975-01-01

    1. The Karyopyknotic index of the palatal and lingual mucosa is increased in female reverse smokers when compared to non-smoking females. 2. The Karyopyknotic index of the buccal mucosa did not show any change in female reverse smokers when compared to non-smoking females. 3. The Karyopyknotic index did not show any change with age in the non-smoking females. 4. Very few cases show epithelial atypia in palatal smears from female reverse smokers having stomatitis nicotina.

  3. Evaluation of Denture Stomatitis in Croatian Adult Population

    OpenAIRE

    Ćelić, R.; Knezović Zlatarić, D.; Baučić, I.

    2001-01-01

    Denture stomatitis (DS) is often found under the removable partial dentures (RPDs). There are many factors influencing it, such as patient’s age and gender, smoking habits, denture age, denture material, denture wearing habits, denture hygiene habits, oral hygiene instruction, denture cleanness and denture plaque accumulation. The aim of this study was to find out the influence these factors have on the prevalence of DS under RPDs and complete dentures (CDs). A total of 200 pat...

  4. Physiology and Regulation of Calcium Channels in Stomatal Guard Cells

    Energy Technology Data Exchange (ETDEWEB)

    Schroeder, Julian I.

    2007-05-02

    Stomatal pores in the epidermis of leaves regulate the diffusion of CO2 into leaves for photosynthetic carbon fixation and control water loss of plants during drought periods. Guard cells sense CO2, water status, light and other environmental conditions to regulate stomatal apertures for optimization of CO2 intake and plant growth under drought stress. The cytosolic second messenger calcium contributes to stomatal movements by transducing signals and regulating ion channels in guard cells. Studies suggest that both plasma membrane Ca2+ influx channels and vacuolar/organellar Ca2+ release channels contribute to ABA-induced Ca2+ elevations in guard cells. Recent research in the P.I.'s laboratory has led to identification of a novel major cation-selective Ca2+-permeable influx channel (Ica) in the plasma membrane of Arabidopsis guard cells. These advances will allow detailed characterization of Ica plasma membrane Ca2+ influx channels in guard cells. The long term goal of this research project is to gain a first detailed characterization of these novel plasma membrane Ca2+-permeable channel currents in Arabidopsis guard cells. The proposed research will investigate the hypothesis that Ica represents an important Ca2+ influx pathway for ABA and CO2 signal transduction in Arabidopsis guard cells. These studies will lead to elucidation of key signal transduction mechanisms by which plants balance CO2 influx into leaves and transpirational water loss and may contribute to future strategies for manipulating gas exchange for improved growth of crop plants and for biomass production.

  5. Silver nitrate cauterization: a treatment option for aphthous stomatitis.

    Science.gov (United States)

    Soylu Özler, Gül

    2014-07-01

    In this study we compared silver nitrate cautery with placebo to assess the effect of silver nitrate cautery in aphthous stomatitis for pain relief and healing time. In this study, sixty-five patients with aphthous stomatitis were assessed. Silver nitrate sticks were used in group A (treatment group) and placebo sticks were used for group B (control group). Change in the severity of pain, change in the size of the ulcers, healing time, side effects of the procedure were assessed. Although the mean value of pain scores before the procedure was similar in both of the groups, there were statistically significant differences between two groups after the procedure on the first to the seventh day. On the seventh day after the procedure, the ulcers were completely reepithelialized in 21 patients (60%) in the treatment group and in 10 patients (32%) in the placebo group. The difference was statistically significant (p aphthous stomatitis. Also this treatment shortens the healing time of ulcers. Copyright © 2013 European Association for Cranio-Maxillo-Facial Surgery. Published by Elsevier Ltd. All rights reserved.

  6. Oral mucosa and therapy of recurrent aphthous stomatitis.

    Science.gov (United States)

    Landová, Hana; Daněk, Zdeněk; Gajdziok, Jan; Vetchý, David; Stembírek, Jan

    2013-02-01

    Oral mucosa is one of the specific surfaces of the human body, which is permanently exposed to external factors related with food intake, breathing and speaking processes, which can lead to the onset of some problems. Disorders of the oral mucosa are a group of diseases, affecting, in the course of life, the majority of the population. Many of the oral mucosa ailments are manifested by lesions. Recurrent aphthous stomatitis (RAS) is the most common of these diseases. Despite much clinical and research attention, its causes remain poorly understood and treatment is only symptomatic. RAS is reported to affect up to 25% of the population worldwide. Topical or systemic therapy (corticosteroids, antiseptics, anti-inflamatory drugs, immunomodulating agents, etc.) can be used for treatment of RAS-associated symptoms. In general, topical therapy should be preferred due to the smaller drug load of the organism. In both cases, the active substance has to be in suitable dosage form. Recently, besides the conventional ways of application (rinses), the main disadvantage of which is the short time of resistance in the oral cavity, mucoadhesive dosage forms are used. The aim of this article is to give a theoretical overview of the oral mucosa topic and its most frequent disease - recurrent aphthous stomatitis in terms of various types of the disease classification, diagnosis and therapy, and in terms of the usage of various types of active substances and medical forms. oral mucosa recurrent aphthous stomatitis therapy mucoadhesive dosage forms.

  7. Ozone exposure and stomatal sluggishness in different plant physiognomic classes

    Energy Technology Data Exchange (ETDEWEB)

    Paoletti, Elena, E-mail: e.paoletti@ipp.cnr.i [IPP-CNR, Via Madonna del Piano 10, I-50019 Sesto Fiorentino, Florence (Italy); Grulke, Nancy E. [US Forest Service, 4955 Canyon Crest Drive, Riverside, CA 92507 (United States)

    2010-08-15

    Gas exchange responses to static and variable light were tested in three species: snap bean (Phaseolus vulgaris, two cultivars), California black oak (Quercus kelloggii), and blue oak (Q. douglasii). The effects of 1-month (snap beans) and 2-month (oaks) O{sub 3} (ozone) exposure (70 ppb over 8 h per day in open-top chambers) were investigated. A delay in stomatal responses (i.e., 'sluggish' responses) to variable light was found to be both an effect of O{sub 3} exposure and a reason for increased O{sub 3} sensitivity in snap bean cultivars, as it implied higher O{sub 3} uptake during times of disequilibrium. Sluggishness increased the time to open (thus limiting CO{sub 2} uptake) and close stomata (thus increasing transpirational water loss) after abrupt changes in light level. Similar responses were shown by snap beans and oaks, suggesting that O{sub 3}-induced stomatal sluggishness is a common trait among different plant physiognomic classes. - Sluggish stomatal responses are suggested to be both an effect of O{sub 3} exposure and a reason of increased O{sub 3} sensitivity in plants.

  8. Mapping the distribution of vesicular textures on silicic lavas using the Thermal Infrared Multispectral Scanner

    Science.gov (United States)

    Ondrusek, Jaime; Christensen, Philip R.; Fink, Jonathan H.

    1993-01-01

    To investigate the effect of vesicularity on TIMS (Thermal Infrared Multispectral Scanner) imagery independent of chemical variations, we studied a large rhyolitic flow of uniform composition but textural heterogeneity. The imagery was recalibrated so that the digital number values for a lake in the scene matched a calculated ideal spectrum for water. TIMS spectra for the lava show useful differences in coarsely and finely vesicular pumice data, particularly in TIMS bands 3 and 4. Images generated by ratioing these bands accurately map out those areas known from field studies to be coarsely vesicular pumice. These texture-related emissivity variations are probably due to the larger vesicles being relatively deeper and separated by smaller septa leaving less smooth glass available to give the characteristic emission of the lava. In studies of inaccessible lava flows (as on Mars) areas of coarsely vesicular pumice must be identified and avoided before chemical variations can be interpreted. Remotely determined distributions of vesicular and glassy textures can also be related to the volatile contents and potential hazards associated with the emplacement of silicic lava flows on Earth.

  9. Vesicular trafficking of immune mediators in human eosinophils revealed by immunoelectron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Melo, Rossana C.N., E-mail: rossana.melo@ufjf.edu.br [Laboratory of Cellular Biology, Department of Biology, ICB, Federal University of Juiz de Fora, UFJF, Rua José Lourenço Kelmer, Juiz de Fora, MG 36036-900 (Brazil); Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, 330 Brookline Avenue, CLS 943, Boston, MA 02215 (United States); Weller, Peter F. [Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, 330 Brookline Avenue, CLS 943, Boston, MA 02215 (United States)

    2016-10-01

    Electron microscopy (EM)-based techniques are mostly responsible for our current view of cell morphology at the subcellular level and continue to play an essential role in biological research. In cells from the immune system, such as eosinophils, EM has helped to understand how cells package and release mediators involved in immune responses. Ultrastructural investigations of human eosinophils enabled visualization of secretory processes in detail and identification of a robust, vesicular trafficking essential for the secretion of immune mediators via a non-classical secretory pathway associated with secretory (specific) granules. This vesicular system is mainly organized as large tubular-vesicular carriers (Eosinophil Sombrero Vesicles – EoSVs) actively formed in response to cell activation and provides a sophisticated structural mechanism for delivery of granule-stored mediators. In this review, we highlight the application of EM techniques to recognize pools of immune mediators at vesicular compartments and to understand the complex secretory pathway within human eosinophils involved in inflammatory and allergic responses. - Highlights: • Application of EM to understand the complex secretory pathway in human eosinophils. • EM techniques reveal an active vesicular system associated with secretory granules. • Tubular vesicles are involved in the transport of granule-derived immune mediators.

  10. Vesicular exanthema of swine virus: isolation and serotyping of field samples.

    Science.gov (United States)

    Edwards, J F; Yedloutschnig, R J; Dardiri, A H; Callis, J J

    1987-01-01

    Virus isolation was attempted from 262 field samples of vesicular material collected during the outbreaks of vesicular exanthema of swine in the U.S.A. from 1952-54. Using primary swine kidney culture, viral cytopathogenic agents were isolated from 76.3% of the samples. However, an overall recovery rate of 82.1% was obtained after samples negative in tissue culture were inoculated intradermally in susceptible swine. All vesicular exanthema of swine virus isolates were identified as serotype B51 using complement fixation and serum neutralization tests. Two isolates did not react with antisera to known vesicular agents of swine and failed to produce vesicles or clinical signs of disease upon inoculation in swine. One vesicular exanthema of swine virus isolate from tissue of equine origin was pathogenic for swine but produced limited vesiculation at the site of intradermalingual inoculation in the tongue of a pony infected experimentally. Type B51 virus was reisolated from lesions produced in the pony and the pony became seropositive for virus type B51. PMID:3651889

  11. Moderate water stress causes different stomatal and non-stomatal changes in the photosynthetic functioning of Phaseolus vulgaris L. genotypes.

    Science.gov (United States)

    Ramalho, J C; Zlatev, Z S; Leitão, A E; Pais, I P; Fortunato, A S; Lidon, F C

    2014-01-01

    The impact of moderate water deficit on the photosynthetic apparatus of three Phaseolus vulgaris L. cultivars, Plovdiv 10 (P10), Dobrudjanski Ran (DR) and Prelom (Prel), was investigated. Water shortage had less impact on leaf hydration, RWC (predawn and midday) and predawn water potential in Prel. RWC and Ψ(p) were more reduced in P10, while there was no osmotic adjustment in any cultivar. Although drought drastically reduced stomatal opening in P10 and DR, reduced A(max) indicated non-stomatal limitations that contributed to the negligible P(n). These limitations were on potential thylakoid electron transport rates of PSI and II, pointing to photosystem functioning as a major limiting step in photosynthesis. This agrees with decreases in actual photochemical efficiency of PSII (F(v)'/F(m)'), quantum yield of photosynthetic non-cyclic electron transport (ϕ(e)) and energy-driven photochemical events (q(P)), although the impact on these parameters would also include down-regulation processes. When compared to DR, Prel retained a higher functional state of the photosynthetic machinery, justifying reduced need for photoprotective mechanisms (non-photochemical quenching, zeaxanthin, lutein, β-carotene) and maintenance of the balance between energy capture and dissipative pigments. The highest increases in fructose, glucose, arabinose and sorbitol in Prel might be related to tolerance to a lower oxidative state. All cultivars had reduced A(max) due to daytime stomatal closure in well-watered conditions. Under moderate drought, Prel had highest tolerance, higher leaf hydration and maintenance of important photochemical use of energy. However, water shortage caused appreciable non-stomatal limitations to photosynthesis linked to regulation/imbalance at the metabolic level (and growth) in all cultivars. This included damage, as reflected in decreased potential photosystem functioning, pointing to higher sensitivity of photosynthesis to drought than is commonly assumed

  12. Diagnosis and management of recurrent herpetiform stomatitis and Behçet syndrome like recurrent aphthous stomatitis herpetiform type

    OpenAIRE

    Endah Ayu Tri Wulandari; Gus Permana Subita

    2008-01-01

    Recurrent Aphthous Stomatitis (RAS) is a common inflammatory condition of the oral mucosa. The aetiology of RAS remains unclear, yet there are several predisposing factors which could be involved in the onset of the lesion. The herpetiform type of RAS appeared to be similar to recurrent oral Herpes Simplex infection and also could be part of Behçet Syndrome. This case report discussed a patient suffering from a herpetiform type of RAS with its clinical appearance resembling recurrent oral Her...

  13. Quantitative Chemical Measurements of Vesicular Transmitters with Electrochemical Cytometry.

    Science.gov (United States)

    Li, Xianchan; Dunevall, Johan; Ewing, Andrew G

    2016-10-18

    Electrochemical cytometry adds a new dimension to our ability to study the chemistry and chemical storage of transmitter molecules stored in nanometer vesicles. The approach involves the adsorption and subsequent rupture of vesicles on an electrode surface during which the electroactive contents are quantitatively oxidized (or reduced). The measured current allows us to count the number of molecules in the vesicles using Faraday's law and to correlate this to the amount of molecules released when single exocytosis events take place at communicating cells. The original format for this method involved a capillary electrophoresis separation step to singly address each vesicle, but we have more recently discovered that cellular vesicles tend to adsorb to carbon electrodes and spontaneously as well as stochastically rupture to give mostly single vesicle events. This approach, called impact electrochemical cytometry, even though the impact is perhaps not the important part of this process, has been studied and the vesicle rupture appears to be at the interface between the vesicle and the electrode and is probably driven by electroporation. The pore size and rate of content electrolysis are a function of the pore diameter and the presence of a protein core in the vesicles. In model liposomes with no protein, events appear extremely rapidly as the soft nanoparticles impact the electrode and the contents are oxidized. It appears that the proteins decorating the surface of the vesicle are important in maintaining a gap from the electrode and when this gap is closed electroporation takes place. Models of the event response times suggest the pores formed are small enough so we can carry out these measurements at nanotip electrodes and we have used this to quantify the vesicle content in living cells in a mode we call intracellular impact electrochemical cytometry. The development of electrochemical cytometry allows comparison between vesicle content and vesicular release and

  14. CO2 sensing and CO2 regulation of stomatal conductance: advances and open questions

    Science.gov (United States)

    Engineer, Cawas; Hashimoto-Sugimoto, Mimi; Negi, Juntaro; Israelsson-Nordstrom, Maria; Azoulay-Shemer, Tamar; Rappel, Wouter-Jan; Iba, Koh; Schroeder, Julian

    2015-01-01

    Guard cells form epidermal stomatal gas exchange valves in plants and regulate the aperture of stomatal pores in response to changes in the carbon dioxide (CO2) concentration in leaves. Moreover, the development of stomata is repressed by elevated CO2 in diverse plant species. Evidence suggests that plants can sense CO2 concentration changes via guard cells and via mesophyll tissues in mediating stomatal movements. We review new discoveries and open questions on mechanisms mediating CO2-regulated stomatal movements and CO2 modulation of stomatal development, which together function in CO2-regulation of stomatal conductance and gas exchange in plants. Research in this area is timely in light of the necessity of selecting and developing crop cultivars which perform better in a shifting climate. PMID:26482956

  15. Effect of SO/sub 2/ on stomatal aperture and sulfur uptake of woody angiosperm seedlings

    Energy Technology Data Exchange (ETDEWEB)

    Noland, T.L.; Kozlowski, T.T.

    1979-01-01

    Effects of SO/sub 2/ pollution on stomatal aperture and sulfur uptake varied with SO/sub 2/ dosage and plant species. Fumigation of Ulmus americana L. seedlings with 1 ppm SO/sub 2/ for 8 h inhibited stomatal closure and fumigation with 2 ppm SO/sub 2/ for 12 h induced stomatal closure. Sulfur uptake of fumigated Ulmus americana seedlings depended on stomatal aperture and was much higher in the light than in the dark. Fumigation of water-stressed Ginkgo biloba L. seedlings with 2 ppm SO/sub 2/ for 6.5 h tended to prevent stomatal closure. However, the effects of SO/sub 2/ on stomatal aperture were modulated and often overridden by environmental stresses such as low light intensity and drought.

  16. Stomatitis-Related Pain in Women with Breast Cancer Undergoing Autologous Hematopoietic Stem Cell Transplant

    OpenAIRE

    Fall-Dickson, Jane M.; Mock, Victoria; Berk, Ronald A.; Grimm, Patricia M.; Davidson, Nancy; Gaston-Johansson, Fannie

    2008-01-01

    The purpose of this cross-sectional, correlational study was to describe stomatitis-related pain in women with breast cancer undergoing autologous hematopoietic stem cell transplant. Hypotheses tested were that significant, positive relationships would exist between oral pain and stomatitis, state anxiety, depression, and alteration in swallowing. Stomatitis, sensory dimension of oral pain, and state anxiety were hypothesized to most accurately predict oral pain overall intensity. Thirty-two ...

  17. Responses of leaf stomatal density to water status and its relationship with photosynthesis in a grass

    OpenAIRE

    Xu, Zhenzhu; Zhou, Guangsheng

    2008-01-01

    Responses of plant leaf stomatal conductance and photosynthesis to water deficit have been extensively reported; however, little is known concerning the relationships of stomatal density with regard to water status and gas exchange. The responses of stomatal density to leaf water status were determined, and correlation with specific leaf area (SLA) in a photosynthetic study of a perennial grass, Leymus chinensis, subjected to different soil moisture contents. Moderate water deficits had posit...

  18. Évaluation des caractéristiques des stomates chez le palmier à huile ...

    African Journals Online (AJOL)

    Les résultats ont montré que, sur la face inférieure des feuilles, la densité stomatique est plus importante chez le matériel adulte 50,32 stomates par mm2 que chez le matériel jeune 23,88 stomates par mm2 et, sur la face supérieure des feuilles, cette situation s'inverse 4,77 stomates par mm2 contre 9,21 stomates par mm2.

  19. Crosstalk among hormones and signaling networks during stomatal development in Arabidopsis hypocotyls

    Directory of Open Access Journals (Sweden)

    Laura Serna

    2016-09-01

    Full Text Available During development, signaling networks specify stomatal cell fate and patterning in response to phytohormones. A number of studies in the past few years have revealed that brassinosteroids repress the signaling pathway that inactivates SPEECHLESS (SPCH, promoting stomatal cell fate determination in the hypocotyl. These plant hormones also control stomatal patterning specification by regulating genes in the TTG/BHLHs/MYBs/GL2 network. Gibberellins, like brassinosteroids, promote stomatal formation in the embryonic stem, which suggests that their signaling pathways may converge. These phytohormones also regulate LLM-domain B-GATA factors. The involvement of these factors as positive regulators of stomatal formation, which function upstream of SPCH, suggests that the brassinosteroid and gibberellin signaling pathways may converge to control stomatal cell fate specification. In addition, the leucine-rich repeat-containing receptor-like protein TOO MANY MOUTHS acts later than these hormones in the cell division sequence that triggers stomatal formation, and it also appears to control stomatal initiation in response to brassinosteroids. The emerging picture suggests that crosstalk among hormones and signaling networks guides stomatal cell fate determination and patterning in the hypocotyl.

  20. Evolutionary association of stomatal traits with leaf vein density in Paphiopedilum, Orchidaceae.

    Directory of Open Access Journals (Sweden)

    Shi-Bao Zhang

    Full Text Available BACKGROUND: Both leaf attributes and stomatal traits are linked to water economy in land plants. However, it is unclear whether these two components are associated evolutionarily. METHODOLOGY/PRINCIPAL FINDINGS: In characterizing the possible effect of phylogeny on leaf attributes and stomatal traits, we hypothesized that a correlated evolution exists between the two. Using a phylogenetic comparative method, we analyzed 14 leaf attributes and stomatal traits for 17 species in Paphiopedilum. Stomatal length (SL, stomatal area (SA, upper cuticular thickness (UCT, and total cuticular thickness (TCT showed strong phylogenetic conservatism whereas stomatal density (SD and stomatal index (SI were significantly convergent. Leaf vein density was correlated with SL and SD whether or not phylogeny was considered. The lower epidermal thickness (LET was correlated positively with SL, SA, and stomatal width but negatively with SD when phylogeny was not considered. When this phylogenetic influence was factored in, only the significant correlation between SL and LET remained. CONCLUSION/SIGNIFICANCE: Our results support the hypothesis for correlated evolution between stomatal traits and vein density in Paphiopedilum. However, they do not provide evidence for an evolutionary association between stomata and leaf thickness. These findings lend insight into the evolution of traits related to water economy for orchids under natural selection.

  1. Stomatal Blue Light Response Is Present in Early Vascular Plants1[OPEN

    Science.gov (United States)

    Doi, Michio; Kitagawa, Yuki; Shimazaki, Ken-ichiro

    2015-01-01

    Light is a major environmental factor required for stomatal opening. Blue light (BL) induces stomatal opening in higher plants as a signal under the photosynthetic active radiation. The stomatal BL response is not present in the fern species of Polypodiopsida. The acquisition of a stomatal BL response might provide competitive advantages in both the uptake of CO2 and prevention of water loss with the ability to rapidly open and close stomata. We surveyed the stomatal opening in response to strong red light (RL) and weak BL under the RL with gas exchange technique in a diverse selection of plant species from euphyllophytes, including spermatophytes and monilophytes, to lycophytes. We showed the presence of RL-induced stomatal opening in most of these species and found that the BL responses operated in all euphyllophytes except Polypodiopsida. We also confirmed that the stomatal opening in lycophytes, the early vascular plants, is driven by plasma membrane proton-translocating adenosine triphosphatase and K+ accumulation in guard cells, which is the same mechanism operating in stomata of angiosperms. These results suggest that the early vascular plants respond to both RL and BL and actively regulate stomatal aperture. We also found three plant species that absolutely require BL for both stomatal opening and photosynthetic CO2 fixation, including a gymnosperm, C. revoluta, and the ferns Equisetum hyemale and Psilotum nudum. PMID:26307440

  2. Evolutionary association of stomatal traits with leaf vein density in Paphiopedilum, Orchidaceae.

    Science.gov (United States)

    Zhang, Shi-Bao; Guan, Zhi-Jie; Sun, Mei; Zhang, Juan-Juan; Cao, Kun-Fang; Hu, Hong

    2012-01-01

    Both leaf attributes and stomatal traits are linked to water economy in land plants. However, it is unclear whether these two components are associated evolutionarily. In characterizing the possible effect of phylogeny on leaf attributes and stomatal traits, we hypothesized that a correlated evolution exists between the two. Using a phylogenetic comparative method, we analyzed 14 leaf attributes and stomatal traits for 17 species in Paphiopedilum. Stomatal length (SL), stomatal area (SA), upper cuticular thickness (UCT), and total cuticular thickness (TCT) showed strong phylogenetic conservatism whereas stomatal density (SD) and stomatal index (SI) were significantly convergent. Leaf vein density was correlated with SL and SD whether or not phylogeny was considered. The lower epidermal thickness (LET) was correlated positively with SL, SA, and stomatal width but negatively with SD when phylogeny was not considered. When this phylogenetic influence was factored in, only the significant correlation between SL and LET remained. Our results support the hypothesis for correlated evolution between stomatal traits and vein density in Paphiopedilum. However, they do not provide evidence for an evolutionary association between stomata and leaf thickness. These findings lend insight into the evolution of traits related to water economy for orchids under natural selection.

  3. Historia natural del virus de la estomatitis vesicular en zonas enzoóticas de Antioquia

    OpenAIRE

    John Arboleda; Andrés Londoño; Víctor Quiroz; Carlos Trujillo

    2003-01-01

    La Estomatitis Vesicular (EV) es una enfermedad producida
    por el virus de la Estomatitis Vesicular, serotipos New Jersey (VSV-NJ) e Indiana (VSV-IN), afecta bovinos y equinos, porcinos y causa infección natural en humanos, principalmente granjeros, ordeñadores y personal de laboratorio.
    Se caracteriza por producir vesículas en las membranas mucosas
    de la boca (epitelio de la lengua y el paladar), bandas coronarias,
    pezones y tejidos blandos...

  4. Guard cell photosynthesis is critical for stomatal turgor production, yet does not directly mediate CO2- and ABA-induced stomatal closing

    Science.gov (United States)

    Azoulay-Shemer, Tamar; Palomares, Axxell; Bagheri, Andish; Israelsson-Nordstrom, Maria; Engineer, Cawas B.; Bargmann, Bastiaan O.R.; Stephan, Aaron B.; Schroeder, Julian I.

    2015-01-01

    SUMMARY Stomata mediate gas exchange between the inter-cellular spaces of leaves and the atmosphere. CO2 levels in leaves (Ci) are determined by respiration, photosynthesis, stomatal conductance and atmospheric [CO2]. [CO2] in leaves mediates stomatal movements. The role of guard-cell photosynthesis in stomatal conductance responses is a matter of debate, and genetic approaches are needed. We have generated transgenic Arabidopsis plants that are chlorophyll-deficient in guard cells only, expressing a constitutively active chlorophyllase in a guard-cell specific enhancer trap-line. Our data show that more than 90% of guard cells were chlorophyll-deficient. Interestingly, approximately ~ 45% of stomata had an unusual, previously not-described, morphology of thin-shaped chlorophyll-less stomata. Nevertheless, stomatal size, stomatal index, plant morphology, and whole-leaf photosynthetic parameters (PSII, qP, qN, FV′/FM′) were comparable to wild-type plants. Time-resolved intact leaf gas exchange analyses showed a reduction in stomatal conductance and carbon assimilation rates of the transgenic plants. Normalization of CO2 responses showed that stomata of transgenic plants respond to [CO2] shifts. Detailed stomatal aperture measurements of normal kidney-shaped stomata, which lack chlorophyll, showed stomatal closing responses to [CO2] elevation and abscisic acid (ABA), while thin-shaped stomata were continuously closed. Our present findings show that stomatal movement responses to [CO2] and ABA are functional in guard cells that lack chlorophyll. These data suggest that guard-cell CO2 and ABA signal transduction are not directly modulated by guard-cell photosynthesis/electron transport. Moreover, the finding that chlorophyll-less stomata cause a “deflated” thin-shaped phenotype, suggests that photosynthesis in guard cells is critical for energization and guard-cell turgor production. PMID:26096271

  5. Guard cell photosynthesis is critical for stomatal turgor production, yet does not directly mediate CO2 - and ABA-induced stomatal closing.

    Science.gov (United States)

    Azoulay-Shemer, Tamar; Palomares, Axxell; Bagheri, Andisheh; Israelsson-Nordstrom, Maria; Engineer, Cawas B; Bargmann, Bastiaan O R; Stephan, Aaron B; Schroeder, Julian I

    2015-08-01

    Stomata mediate gas exchange between the inter-cellular spaces of leaves and the atmosphere. CO2 levels in leaves (Ci) are determined by respiration, photosynthesis, stomatal conductance and atmospheric [CO2 ]. [CO2 ] in leaves mediates stomatal movements. The role of guard cell photosynthesis in stomatal conductance responses is a matter of debate, and genetic approaches are needed. We have generated transgenic Arabidopsis plants that are chlorophyll-deficient in guard cells only, expressing a constitutively active chlorophyllase in a guard cell specific enhancer trap line. Our data show that more than 90% of guard cells were chlorophyll-deficient. Interestingly, approximately 45% of stomata had an unusual, previously not-described, morphology of thin-shaped chlorophyll-less stomata. Nevertheless, stomatal size, stomatal index, plant morphology, and whole-leaf photosynthetic parameters (PSII, qP, qN, FV '/FM' ) were comparable with wild-type plants. Time-resolved intact leaf gas-exchange analyses showed a reduction in stomatal conductance and CO2 -assimilation rates of the transgenic plants. Normalization of CO2 responses showed that stomata of transgenic plants respond to [CO2 ] shifts. Detailed stomatal aperture measurements of normal kidney-shaped stomata, which lack chlorophyll, showed stomatal closing responses to [CO2 ] elevation and abscisic acid (ABA), while thin-shaped stomata were continuously closed. Our present findings show that stomatal movement responses to [CO2 ] and ABA are functional in guard cells that lack chlorophyll. These data suggest that guard cell CO2 and ABA signal transduction are not directly modulated by guard cell photosynthesis/electron transport. Moreover, the finding that chlorophyll-less stomata cause a 'deflated' thin-shaped phenotype, suggests that photosynthesis in guard cells is critical for energization and guard cell turgor production. © 2015 The Authors The Plant Journal © 2015 John Wiley & Sons Ltd.

  6. A putative vesicular transporter expressed in Drosophila mushroom bodies that mediates sexual behavior may define a novel neurotransmitter system

    Science.gov (United States)

    Brooks, Elizabeth S.; Greer, Christina L.; Romero-Calderón, Rafael; Serway, Christine N.; Grygoruk, Anna; Haimovitz, Jasmine M.; Nguyen, Bac T.; Najibi, Rod; Tabone, Christopher J.; de Belle, J. Steven; Krantz, David E.

    2011-01-01

    Summary Storage and release of classical and amino acid neurotransmitters requires vesicular transporters. Some neurons lack known vesicular transporters, suggesting additional neurotransmitter systems remain unidentified. Insect mushroom bodies (MBs) are critical for several behaviors, including learning, but the neurotransmitters released by the intrinsic Kenyon cells (KCs) remain unknown. Likewise, KCs do not express a known vesicular transporter. We report the identification of a novel Drosophila gene portabella (prt) that is structurally similar to known vesicular transporters. Both larval and adult brains express PRT in the KCs of the MBs. Additional PRT cells project to the central complex and optic ganglia. prt mutation causes an olfactory learning deficit and an unusual defect in the male’s position during copulation that is rescued by expression in KCs. Since prt is expressed in neurons that lack other known vesicular transporters or neurotransmitters, it may define a previously unknown neurotransmitter system responsible for sexual behavior and a component of olfactory learning. PMID:22017990

  7. The beneficial effect of dual inoculation of vesicular-arbuscular mycorrhizae + rhizobium on growth of white clover

    Directory of Open Access Journals (Sweden)

    Lin, XG.

    1993-01-01

    Full Text Available Investigation on the effect of phosphorus on vesicular-arbuscular mycorrhizal infection, and dual inoculation of vesicular-arbuscular mycorrhizae + rhizobium on growth of white clover under field microplots and pot experiments was conducted on fluvo-aquic soils of semi-arid region in north China. The results showed that 60 kg P205 ha in form of superphosphate was the most favorable phosphorus level for vesicular-arbuscular mycorrhizal infection ; mycorrhizal infection, nodulation, dry weight of shoots and roots, total uptake of nitrogen, phosphorus and other elements, the final yields and recovery of phosphorus of white clover were significantly increased by vesicular-arbuscular mycorrhizal inoculation and dual inoculation with vesicular-arbuscular mycorrhizal fungi and rhizobium. The highest response of inoculation was obtained by adding fertilizer phosphorus at the level of 60 kg P205 ha in form of superphosphate.

  8. Association between serotonin transporter gene polymorphism and recurrent aphthous stomatitis.

    Science.gov (United States)

    Manchanda, Aastha; Iyengar, Asha R; Patil, Seema

    2016-01-01

    Anxiety-related traits have been attributed to sequence variability in the genes coding for serotonin transmission in  the brain. Two alleles, termed long (L) and short (S) differing by 44 base pairs, are found in a polymorphism identified in the promoter region of serotonin transporter gene. The presence of the short allele  and SS and LS genotypes is found to be associated with the reduced expression of this gene decreasing the uptake of serotonin in the brain leading to various anxiety-related traits. Recurrent aphthous stomatitis (RAS) is an oral mucosal disease with varied etiology including the presence of stress, anxiety, and genetic influences. The present study aimed to determine this serotonin transporter gene polymorphism in patients with RAS and compare it with normal individuals. This study included 20 subjects with various forms of RAS and 20 normal healthy age- and gender-matched individuals. Desquamated oral mucosal cells were collected for DNA extraction and subjected to polymerase chain reaction for studying insertion/deletion in the 5-HTT gene-linked polymorphic region. Cross tabulations followed by Chi-square tests were performed to compare the significance of findings, P aphthous stomatitis (60%) and controls (40%). The total percentage of LS and SS genotypes and the frequency of S allele were found to be higher in the subjects with aphthous stomatitis as compared to the control group although a statistically significant correlation could not be established, P = 0.144 and 0.371, respectively. Within the limitations of this study, occurrence of RAS was not found to be associated with polymorphic promoter region in serotonin transporter gene.

  9. Modelling annual pasture dynamics: Application to stomatal ozone deposition

    Science.gov (United States)

    González-Fernández, Ignacio; Bermejo, Victoria; Elvira, Susana; Sanz, Javier; Gimeno, Benjamín S.; Alonso, Rocío

    2010-07-01

    Modelling ozone (O 3) deposition for impact risk assessment is still poorly developed for herbaceous vegetation, particularly for Mediterranean annual pastures. High inter-annual climatic variability in the Mediterranean area makes it difficult to develop models characterizing gas exchange behaviour and air pollutant absorption suitable for risk assessment. This paper presents a new model to estimate stomatal conductance (g s) of Trifolium subterraneum, a characteristic species of dehesa pastures. The MEDPAS (MEDiterranean PAStures) model couples 3 modules estimating soil water content (SWC), vegetation growth and gs. The gs module is a reparameterized version of the stomatal component of the EMEP DO 3SE O 3 deposition model. The MEDPAS model was applied to two contrasting years representing typical dry and humid springs respectively and with different O 3 exposures. The MEDPAS model reproduced realistically the gs seasonal and inter-annual variations observed in the field. SWC was identified as the major driver of differences across years. Despite the higher O 3 exposure in the dry year, meteorological conditions favoured 2.1 times higher gs and 56 day longer growing season in the humid year compared to the dry year. This resulted in higher ozone fluxes absorbed by T. subterraneum in the humid year. High inter-family variability was found in gas exchange rates, therefore limiting the relevance of single species O 3 deposition flux modelling for dehesa pastures. Stomatal conductance dynamics at the canopy level need to be considered for more accurate O 3 flux modelling for present and future climate scenarios in the Mediterranean area.

  10. STOMATITIS AFTOSA REKUREN DAN GANGGUAN FUNGSI OVARIUM (Laporan Kasus

    Directory of Open Access Journals (Sweden)

    Farida Soetiarto

    2015-08-01

    Full Text Available Recurrent aphthous stomatitis (RAS is a common disease of undefine etiology. Many studies have been conducted but the result remain unsatisfied. Fortunately some predisposing factors have been identified and one of predisposing factors is hormonal imbalance. The cases report showed that RAS in ovary disorder and disappear after ovary was amputated, and a case of infertility. IN case of RAS with hormone imbalance as predisposing factor, it is suggested to consider the condition of ovary. This suggestion is based on fact that ovary is producing hormone (estrogen, progesteron whihc influence to oral mucous.

  11. Effects of ozone-induced stomatal closure on ozone uptake and its changes due to leaf age in sun and shade leaves of Siebold's beech

    OpenAIRE

    Hoshika, Yasutomo; Watanabe, Makoto; Inada, Naoki; Koike, Takayoshi

    2015-01-01

    An estimation of stomatal ozone uptake for the assessment of ozone risks in forest trees can be modified by ozone-induced stomatal closure. We thus examined a seasonal course of stomatal conductance in sun and shade leaves of Siebold's beech native to northern Japan (Fagus crenata) grown under free-air ozone exposure. A performance of multiplicative stomatal conductance model was also tested, when considering ozone-induced stomatal closure into the model. Ozone caused stomatal closure in both...

  12. Increase in vesicular hand eczema after house dust mite inhalation provocation : a double-blind, placebo-controlled, cross-over study

    NARCIS (Netherlands)

    Schuttelaar, Marielouise; Coenraads, Pieter Jan; Huizinga, Janneke; De Monchy, Jan G; Vermeulen, Karin M

    BACKGROUND: It is unclear whether the respiratory tract is involved in eliciting or aggravating eczematous lesions in patients with vesicular hand eczema. Objectives. To investigate the effect of inhalation of house dust mite (HDM) on vesicular hand eczema. METHODS: Eighteen patients with vesicular

  13. Detection of three porcine vesicular viruses using multiplex real-time primer-probe energy transfer

    DEFF Research Database (Denmark)

    Rasmussen, Thomas Bruun; Uttenthal, Åse; Aguero, M.

    2006-01-01

    Rapid identification of the etiologic agent in infected animals is important for the control of an outbreak of vesicular disease in livestock. We have in the present study developed a multiplex real-time reverse transcription-PCR, based on primer-probe energy transfer (PriProET), for simultaneous...

  14. Toluene-induced, Ca2+-dependent vesicular catecholamine release in rat PC12 cells

    NARCIS (Netherlands)

    Westerink, R.H.S.|info:eu-repo/dai/nl/239425952; Vijverberg, H.P.M.|info:eu-repo/dai/nl/068856474

    2002-01-01

    Acute effects of toluene on vesicular catecholamine release from intact PC12 phaeochromocytoma cells have been investigated using carbon fiber microelectrode amperometry. The frequency of vesicles released is low under basal conditions and is enhanced by depolarization. Toluene causes an increase in

  15. Altered vesicular glutamate transporter expression in human temporal lobe epilepsy with hippocampal sclerosis

    NARCIS (Netherlands)

    Van Liefferinge, J.; Jensen, C.J.; Albertini, G.; Bentea, E.; Demuyser, T.; Merckx, E.; Aronica, E.; Smolders, I.; Massie, A.

    2015-01-01

    Vesicular glutamate transporters (VGLUTs) are responsible for loading glutamate into synaptic vesicles. Altered VGLUT protein expression has been suggested to affect quantal size and glutamate release under both physiological and pathological conditions. In this study, we investigated mRNA and

  16. Influence of the preparation route on the supramolecular organization of lipids in a vesicular system

    DEFF Research Database (Denmark)

    Elizondo, Elisa; Larsen, Jannik; Hatzakis, Nikos

    2012-01-01

    A confocal fluorescence microscopy-based assay was used for studying the influence of the preparation route on the supramolecular organization of lipids in a vesicular system. In this work, vesicles composed of cholesterol and CTAB (1/1 mol %) or cholesterol and DOPC (2/8 mol %) and incorporating...

  17. [Seminal vesicular cysts associated with renal agenesis, ipsilateral ureter and hemitrigone. Report of a case].

    Science.gov (United States)

    Llopis Mínguez, B; Ferrutxe Frau, J; Moreno Pardo, B; Baixauli Martínez, J M; Moreno Barrachina, E; Rodríguez Hernández, J H

    1979-01-01

    A case is presented of seminal vesicular cyst associated with kidney agenesia, ipsilateral ureter and hemitrigon; this is the 17th case presented in the world literature reviewed. A study is made of all the cases published and the authors recommend deferento-vesiculography as the best means of diagnosis and total excision of the cyst as the most effective treatment.

  18. 9 CFR 94.12 - Pork and pork products from regions where swine vesicular disease exists.

    Science.gov (United States)

    2010-01-01

    ... exists. (a) Swine vesicular disease is considered to exist in all regions of the world except Australia..., Latvia, Lithuania, Luxembourg, Mexico, the Netherlands, New Zealand, Norway, Panama, Poland, Portugal..., the pork or pork products must be moved under Department seals or seals of the U.S. Customs Service...

  19. The development and significance of vesicular-arbuscular mycorrhizas as influenced by agricultural practices

    NARCIS (Netherlands)

    Ruissen, M.A.

    1982-01-01

    The development and significance of vesicular- arbuscular mycorrhizas (VAM) in wheat and potatoes have been studied in relation to various farming systems and agricultural practices. The effects of farming systems on VAM have been observed on three neighbouring experimental farms in the vicinity of

  20. Nanoscale distribution of presynaptic Ca(2+) channels and its impact on vesicular release during development.

    Science.gov (United States)

    Nakamura, Yukihiro; Harada, Harumi; Kamasawa, Naomi; Matsui, Ko; Rothman, Jason S; Shigemoto, Ryuichi; Silver, R Angus; DiGregorio, David A; Takahashi, Tomoyuki

    2015-01-07

    Synaptic efficacy and precision are influenced by the coupling of voltage-gated Ca(2+) channels (VGCCs) to vesicles. But because the topography of VGCCs and their proximity to vesicles is unknown, a quantitative understanding of the determinants of vesicular release at nanometer scale is lacking. To investigate this, we combined freeze-fracture replica immunogold labeling of Cav2.1 channels, local [Ca(2+)] imaging, and patch pipette perfusion of EGTA at the calyx of Held. Between postnatal day 7 and 21, VGCCs formed variable sized clusters and vesicular release became less sensitive to EGTA, whereas fixed Ca(2+) buffer properties remained constant. Experimentally constrained reaction-diffusion simulations suggest that Ca(2+) sensors for vesicular release are located at the perimeter of VGCC clusters (<30 nm) and predict that VGCC number per cluster determines vesicular release probability without altering release time course. This "perimeter release model" provides a unifying framework accounting for developmental changes in both synaptic efficacy and time course. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  1. Les caractéristiques des stomates des feuilles de Ficus benjamina L ...

    African Journals Online (AJOL)

    Objective: The main objective of this study is to assess the potential of Ficus benjamina stomata to be used as indicators of local air pollution. Methodology: Stomatal prints were taken from the species of study in the vicinity of roads, in residential and industrial areas and parks. Density, pore surface and stomatal resistance ...

  2. Clinical importance of celiac disease in patients with recurrent aphthous stomatitis.

    Science.gov (United States)

    Yaşar, Sirin; Yaşar, Bülent; Abut, Evren; Aşiran Serdar, Zehra

    2012-02-01

    Recurrent aphthous stomatitis is a common disease of the oral mucosa that is characterized by recurrent, painful ulcers of unknown etiology. The association between celiac disease and recurrent aphthous stomatitis has been evaluated in several studies, but variable results have been reported. The purpose of this study was to determine the prevalence of celiac disease in patients with recurrent aphthous stomatitis. The study group consisted of 82 patients, all of whom had a history of recurrent aphthous stomatitis. The control group included 82 patients who did not have aphthous stomatitis. Patients were screened for IgA anti-endomysial antibodies, IgG anti-endomysial antibodies, IgA anti-gliadin antibodies, and IgG anti-gliadin antibodies. Patients with positive serology underwent endoscopic biopsies of the duodenal mucosa. Patients in both groups were also questioned regarding gastrointestinal symptoms. One patient (1.2%) out of 82 in the study group was diagnosed with celiac disease by biopsy. Gastroesophageal reflux disease symptoms, heartburn and regurgitation were determined to be of higher incidence in the study group (paphthous stomatitis and celiac disease and that screening recurrent aphthous stomatitis patients for celiac disease has little clinical value. Additionally, regurgitation of gastric acid to the oral cavity may precipitate the formation of aphthous stomatitis.

  3. Familial cases of periodic fever with aphthous stomatitis, pharyngitis, and cervical adenitis syndrome.

    Science.gov (United States)

    Adachi, Masao; Watanabe, Aika; Nishiyama, Atsushi; Oyazato, Yoshinobu; Kamioka, Ichiro; Murase, Masanori; Ishida, Akihito; Sakai, Hidemasa; Nishikomori, Ryuta; Heike, Toshio

    2011-01-01

    We report three familial cases of periodic fever with aphthous stomatitis, pharyngitis, and cervical adenitis syndrome, including a pair of monozygotic twins and their mother. It suggests that periodic fever with aphthous stomatitis, pharyngitis, and cervical adenitis syndrome may have a certain monogenetic background. Copyright © 2011 Mosby, Inc. All rights reserved.

  4. Genetic analysis of differences in stomatal guard cell lengths of bread wheat

    Directory of Open Access Journals (Sweden)

    Наталия Петровна Ламари

    2015-05-01

    Full Text Available Variation in stomatal guard cell length of parental cultivars and its inheritance in F1 and F2 hybrids have been studied after crossing between contrast genotypes of winter wheat (Triticum aestivum L.. Analysis of F2 populations has shown the action of three non-allelic genes in control of stomatal guard cell length of parental cultivars

  5. [The laser therapy and laser acupunture of patients with chronic recurrent aphthous stomatitis].

    Science.gov (United States)

    Mikhaĭlova, R I; Terekhova, N V; Zemskaia, E A; Melkadze, N

    1992-01-01

    Laser therapy and laser acupuncture of the biologically active sites were administered to 24 patients with chronic recurrent aphthous stomatitis. The biologically active sites were selected individually with due consideration for the underlying somatic condition. Good results were achieved in the patients with the fibrous form of chronic aphthous stomatitis. Secretory and serum immunoglobulin levels were monitored over the course of laser treatment.

  6. Stomatal response characteristics of Tradescantia virginiana grown at high relative air humidity

    NARCIS (Netherlands)

    Rezaei Nejad, A.; Meeteren, van U.

    2005-01-01

    Plants produced at high relative air humidity (RH) show poor control of water loss after transferring to low RH, a phenomenon which is thought to be due to their stomatal behaviour. The stomatal anatomy and responses of moderate (55%) and high (90%) RH grown Tradescantia virginiana plants to

  7. Cultivar Differences in the Stomatal Characteristics of Cut Roses Grown at High Relative Humidity

    NARCIS (Netherlands)

    Fanourakis, D.; Tapia, A.; Heuvelink, E.; Pinto De Carvalho, S.M.

    2009-01-01

    High relative air humidity (RH>85%) during cultivation is known to reduce the vase life of cut roses, but the magnitude of such effect is cultivar dependent. The reasons behind this genotypic variation are not yet known. In this study, the stomatal density and stomatal responses to two closing

  8. Moderate salinity improves stomatal functioning in rose plants grown at high relative air humidity

    NARCIS (Netherlands)

    Carvalho, Dália R.A.; Vasconcelos, Marta W.; Lee, Sang; Vreugdenhil, Dick; Heuvelink, Ep; Pinto de Carvalho, Susana

    2017-01-01

    Plants grown at high relative air humidity (RH ≥ 85%) show hampered stomatal closure in response to closing stimuli. We hypothesized that a moderate salinity during growth could trigger a stress response and stimulate stomatal functioning due to an increased leaf abscisic acid concentration ([ABA]).

  9. Canopy Transpiration and Stomatal Responses to Prolonged Drought by a Dominant Desert Species in Central Asia

    Directory of Open Access Journals (Sweden)

    Daxing Gu

    2017-06-01

    Full Text Available In arid and semiarid lands, canopy transpiration and its dynamics depend largely on stomatal sensitivity to drought. In this study, the sap flow of a dominant species, Haloxylon ammodendron growing in Central Asian deserts, was monitored using Granier-type sensors, from which the canopy stomatal conductance was derived. The responses of canopy transpiration and stomatal conductance to environmental variables during the second half of the growing season, when annual prolonged drought occurred, was analyzed for four continuous years, from 2013 to 2016. A soil water content (SWC of 3% was identified as the lower soil water threshold for this species, below which the plant lost the ability for stomatal regulation on water loss and suffered the risk of mortality. Above this threshold, the sensitivity of canopy transpiration to vapor pressure deficit, VPD (K, was linearly correlated with SWC, which mainly resulted from different stomatal behaviors at varying drought intensities. Stomatal sensitivity to VPD (m/Gsref increased linearly with soil moisture deficit, inducing a shift from more anisohydric to a more isohydric stomatal behavior. The flexibility of stomatal behavior regarding soil drought was one key element facilitating the survival of H. ammodendron in such an extreme dry environment.

  10. Comparison of different stomatal conductance algorithms for ozone flux modelling [Proceedings

    Science.gov (United States)

    P. Buker; L. D. Emberson; M. R. Ashmore; G. Gerosa; C. Jacobs; W. J. Massman; J. Muller; N. Nikolov; K. Novak; E. Oksanen; D. De La Torre; J. -P. Tuovinen

    2006-01-01

    The ozone deposition model (D03SE) that has been developed and applied within the EMEP photooxidant model (Emberson et al., 2000, Simpson et al. 2003) currently estimates stomatal ozone flux using a stomatal conductance (gs) model based on the multiplicative algorithm initially developed by Jarvis (1976). This model links gs to environmental and phenological parameters...

  11. Comorbidity of recurrent aphthous stomatitis and polyps ventriculi.

    Science.gov (United States)

    Pokupec, Josipa Sanja Gruden; Lukenda, Dolores Biocina

    2013-03-01

    As it is known, many diseases of gastric system cause changes in the oral cavity, with either pathological findings or subjective impressions. When these changes are of pathological nature, the most common finding is recurrent aphthous stomatitis on the tongue, which emerges as a consequence of gastric diseases. Recurrent aphthous stomatitis is a disorder characterised by recurrent ulcerations limited to the oral mucosa, without any other signs of diseases. According to their clinical form, they may be big, small and hyperform. Etiology of recurrent aphthae is genetic predisposition, systemic diseases (virus, certain vitamin deficiency, gastric disorders), and autoimmune disorder and psychogenesis. The symptoms include a prodromal burning sensation and ulceration emerging within 24-48 hours as round symmetrical lesions inflicting the entire oral cavity except for palate and gingiva. Polyps ventriculi are tumours on the gastric mucosa. They can lie on a broad background or hang on the stem, and may be both individual and clustered at the same time. They are more common with elderly male population. They may have a malignant alteration. According to WHO, they have been classified as hyperplastic and neoplastic polyps. Etiology of polyps is atrophic gastritis or H. pylori.

  12. Triamcinolone Acetonide Oromucoadhesive Paste for Treatment of Aphthous Stomatitis

    Directory of Open Access Journals (Sweden)

    Hamed Hamishehkar

    2015-06-01

    Full Text Available Purpose: The aim of this study was to prepare the optimized oral paste formulation of Triamcinolone acetonide intended to be used in aphtous stomatitis. Methods: Plastibases were prepared using mineral oil and polyethylene (95:5. Oral paste formulations were prepared with different mixtures of three hydrocolloids solids, including gelatin, pectin and sodium carboxymethylcellulose, with different ratios, as well as Plastibase. Long-term and short-term stability of prepared formulations were studied in the case of color and consistency of pastes. Franz diffusion cell and dialysis membrane were employed for release study. Release data were fitted in the kinetic models to find out the mechanism of drug release. Results: Formulation containing 60% plastibase, 3.3% pectin, 6.6% gelatin and 30% carboxymethylcellulose showed desired durability of adhesion, spreadability and rheology property in healthy volunteers and was compared with reference formulation (Adcortyl® in the case of release profile. Although, optimized formulation and Adcortyl followed the Higuchi and first order release kinetics respectively, optimized formulation showed similar release profile to reference formulation. Conclusion: Optimized oral paste formulation of Triamcinolone Acetonide showed similar characteristics with reference formulation and could be used as an effective drug delivery system for the treatment of recurrent aphthous stomatitis.

  13. Periodic fever, aphthous stomatitis, pharyngitis, and adenitis (PFAPA) syndrome.

    Science.gov (United States)

    Ali, Nora S; Sartori-Valinotti, Julio C; Bruce, Alison J

    2016-01-01

    Periodic fever, aphthous stomatitis, pharyngitis, and adenitis (PFAPA) syndrome, the most common periodic disorder of childhood, presents with the cardinal symptoms of periodic fever, aphthous stomatitis, pharyngitis, and adenitis typically before age 5. This review presents the recent literature on PFAPA and summarizes key findings in the pathogenesis, evaluation, and treatment of the disease. Theories surrounding the pathogenesis of PFAPA include a faulty innate immunologic response in conjunction with dysregulated T-cell activation. A potential genetic link is also under consideration. Mediterranean fever (MEFV) gene variants have been implicated and appear to modify disease severity. In individuals with the heterozygous variant, PFAPA episodes are milder and shorter in duration. Diagnostic criteria include the traditional clinical signs, in addition to the following biomarkers: elevated C-reactive protein in the absence of elevated procalcitonin, vitamin D, CD64, mean corpuscular volume, and other nonspecific inflammatory mediators in the absence of an infectious explanation for fever. Treatment of PFAPA includes tonsillectomy, a single dose of corticosteroids, and, most recently, interleukin 1 blockers such as anakinra, rilonacept, and canakinumab. Tonsillectomy remains the only permanent treatment modality. Copyright © 2016 Elsevier Inc. All rights reserved.

  14. Stomatal design principles in synthetic and real leaves.

    Science.gov (United States)

    Zwieniecki, Maciej A; Haaning, Katrine S; Boyce, C Kevin; Jensen, Kaare H

    2016-11-01

    Stomata are portals in plant leaves that control gas exchange for photosynthesis, a process fundamental to life on Earth. Gas fluxes and plant productivity depend on external factors such as light, water and CO2 availability and on the geometrical properties of the stoma pores. The link between stoma geometry and environmental factors has informed a wide range of scientific fields-from agriculture to climate science, where observed variations in stoma size and density are used to infer prehistoric atmospheric CO2 content. However, the physical mechanisms and design principles responsible for major trends in stomatal patterning are not well understood. Here, we use a combination of biomimetic experiments and theory to rationalize the observed changes in stoma geometry. We show that the observed correlations between stoma size and density are consistent with the hypothesis that plants favour efficient use of space and maximum control of dynamic gas conductivity, and that the capacity for gas exchange in plants has remained constant over at least the last 325 Myr. Our analysis provides a new measure to gauge the relative performance of species based on their stomatal characteristics. © 2016 The Author(s).

  15. Denture-Related Stomatitis Is Associated with Endothelial Dysfunction

    Directory of Open Access Journals (Sweden)

    Joanna Maciąg

    2014-01-01

    Full Text Available Oral inflammation, such as periodontitis, can lead to endothelial dysfunction, accelerated atherosclerosis, and vascular dysfunction. The relationship between vascular dysfunction and other common forms of oral infections such as denture-related stomatitis (DRS is unknown. Similar risk factors predispose to both conditions including smoking, diabetes, age, and obesity. Accordingly, we aimed to investigate endothelial function and major vascular disease risk factors in 44 consecutive patients with dentures with clinical and microbiological features of DRS (n=20 and without DRS (n=24. While there was a tendency for higher occurrence of diabetes and smoking, groups did not differ significantly in respect to major vascular disease risk factors. Groups did not differ in main ambulatory blood pressure, total cholesterol, or even CRP. Importantly, flow mediated dilatation (FMD was significantly lower in DRS than in non-DRS subjects, while nitroglycerin induced vasorelaxation (NMD or intima-media thickness (IMT was similar. Interestingly, while triglyceride levels were normal in both groups, they were higher in DRS subjects, although they did not correlate with either FMD or NMD. Conclusions. Denture related stomatitis is associated with endothelial dysfunction in elderly patients with dentures. This is in part related to the fact that diabetes and smoking increase risk of both DRS and cardiovascular disease.

  16. Stomatal Responses to CO2 in Paphiopedilum and Phragmipedium1

    Science.gov (United States)

    Assmann, Sarah M.; Zeiger, Eduardo

    1985-01-01

    A role of the guard cell chloroplasts in the CO2 response of stomata was investigated through a comparison of the leaf gas exchange characteristics of two closely related orchids: Paphiopedilum harrisianum, which lacks guard cell chloroplasts and Phragmipedium longifolium, which has chlorophyllous guard cells. Leaves of both species had an apparent quantum yield for assimilation of about 0.05, with photosynthesis saturating at 0.300 to 0.400 millimoles per square meter per second. CO2 curves were obtained by measuring steady-state assimilation and stomatal conductance under 0.180 or 0.053 millimoles per square meter per second white light, or darkness, at 0 to 400 microliters per liter ambient CO2. The response of assimilation to changes in CO2 was similar in the two species, but the response of conductance was consistently weaker in Paphiopedilum than in Phragmipedium. The data suggest involvement of guard cell chloroplasts in the stomatal response to CO2 and in the coupling of assimilation and conductance in the intact leaf. PMID:16664075

  17. Stomatal development in new leaves is related to the stomatal conductance of mature leaves in poplar (Populus trichocarpaxP. deltoides).

    Science.gov (United States)

    Miyazawa, Shin-Ichi; Livingston, Nigel J; Turpin, David H

    2006-01-01

    In general, stomatal density (SD) decreases when plants are grown at high CO2 concentrations. Recent studies suggest that signals produced from mature leaves regulate the SD of expanding leaves. To determine the underlying driver of these signals in poplar (Populus trichocarpaxP. deltoides) saplings, a cuvette system was used whereby the environment around mature (lower) leaves could be controlled independently of that around developing (upper) leaves. A series of experiments were performed in which the CO2 concentration, vapour pressure deficit (D), and irradiance (Q) around the lower leaves were varied while the (ambient) conditions around the upper leaves were unchanged. The overall objective was to break the nexus between leaf stomatal conductance and transpiration and photosynthesis rates of lower leaves and determine which, if any, of these parameters regulate stomatal development in the upper expanding leaves. SD, stomatal index (SI), and epidermal cell density (ED) were measured on the adaxial and abaxial surfaces of fully expanded upper leaves. SD and SI decreased with increasing lower leaf CO2 concentration (150-780 ppm) at both ambient (1.3-1.6 kPa) and low (0.7-1.0 kPa) D. SD and SI at low D were generally higher than at ambient D. By contrast, ED was relatively insensitive to both vapour pressure and CO2 concentration. When lower leaves were shaded, upper leaf SD, SI, and ED decreased but did not change with varying CO2 concentration. These results suggest that epidermal cell development and stomatal development are regulated by different physiological mechanisms. SI of the upper leaves was positively and highly correlated (r2>0.84) with the stomatal conductance of the lower leaves independent of their net photosynthesis and transpiration rates, suggesting that the stomatal conductance of mature leaves has a regulatory effect on the stomatal development of expanding leaves.

  18. Modeling of Stomatal Conductance for Estimating Ozone Uptake of Fagus crenata Under Experimentally Enhanced Free-air Ozone Exposure

    OpenAIRE

    Hoshika, Yasutomo; Watanabe, Makoto; Inada, Naoki; Koike, Takayoshi

    2012-01-01

    We examined a performance of the multiplicative stomatal conductance model to estimate the stomatal ozone uptake for Fagus crenata. Parameterization of the model was carried out by in-situ measurements in a free-air ozone exposure experiment. The model performed fairly well under ambient conditions, with low ozone concentration. However, the model overestimated stomatal conductance under enhanced ozone condition due to ozone-induced stomatal closure. A revised model that included a parameter ...

  19. Balancing Water Uptake and Loss through the Coordinated Regulation of Stomatal and Root Development

    Science.gov (United States)

    Hepworth, Christopher; Turner, Carla; Landim, Marcela Guimaraes; Cameron, Duncan; Gray, Julie E.

    2016-01-01

    Root development is influenced by nutrient and water availabilities. Plants are able to adjust many attributes of their root in response to environmental signals including the size and shape of the primary root, lateral roots and root hairs. Here we investigated the response of roots to changes in the levels of leaf transpiration associated with altered stomatal frequency. We found that plants with high stomatal density and conductance produce a larger rooting area and as a result have enhanced phosphate uptake capacity whereas plants with low stomatal conductance produce a smaller root. Manipulating the growth environment of plants indicated that enhanced root growth is most likely a result of an increased demand for water rather than phosphate. Plants manipulated to have an increase or reduction in root hair growth show a reduction or increase respectively, in stomatal conductance and density. Our results demonstrate that plants can balance their water uptake and loss through coordinated regulation of both stomatal and root development. PMID:27275842

  20. Combining sap flow and eddy covariance approaches to derive stomatal and non-stomatal O{sub 3} fluxes in a forest stand

    Energy Technology Data Exchange (ETDEWEB)

    Nunn, A.J. [Ecophysiology of Plants, Technische Universitaet Muenchen, Am Hochanger 13, D-85354 Freising- Weihenstephan (Germany); Cieslik, S. [Institute for Environment and Sustainability, Joint Research Center, Ispra (Italy); Metzger, U. [Ecophysiology of Plants, Technische Universitaet Muenchen, Am Hochanger 13, D-85354 Freising- Weihenstephan (Germany); Wieser, G. [Federal Research and Training Centre for Forests, Natural Hazards and Landscape, Dept. of Alpine Timberline Ecophysiology, Rennweg 1, A - 6020 Innsbruck (Austria); Matyssek, R., E-mail: matyssek@wzw.tum.d [Ecophysiology of Plants, Technische Universitaet Muenchen, Am Hochanger 13, D-85354 Freising- Weihenstephan (Germany)

    2010-06-15

    Stomatal O{sub 3} fluxes to a mixed beech/spruce stand (Fagus sylvatica/Picea abies) in Central Europe were determined using two different approaches. The sap flow technique yielded the tree-level transpiration, whereas the eddy covariance method provided the stand-level evapotranspiration. Both data were then converted into stomatal ozone fluxes, exemplifying this novel concept for July 2007. Sap flow-based stomatal O{sub 3} flux was 33% of the total O{sub 3} flux, whereas derivation from evapotranspiration rates in combination with the Penman-Monteith algorithm amounted to 47%. In addition to this proportional difference, the sap flow-based assessment yielded lower levels of stomatal O{sub 3} flux and reflected stomatal regulation rather than O{sub 3} exposure, paralleling the daily courses of canopy conductance for water vapor and eddy covariance-based total stand-level O{sub 3} flux. The demonstrated combination of sap flow and eddy covariance approaches supports the development of O{sub 3} risk assessment in forests from O{sub 3} exposure towards flux-based concepts. - Combined tree sap flow and eddy covariance-based methodologies yield stomatal O{sub 3} flux as 33% in total stand flux.

  1. Does Size Matter? Atmospheric CO2 May Be a Stronger Driver of Stomatal Closing Rate Than Stomatal Size in Taxa That Diversified under Low CO2

    Science.gov (United States)

    Elliott-Kingston, Caroline; Haworth, Matthew; Yearsley, Jon M.; Batke, Sven P.; Lawson, Tracy; McElwain, Jennifer C.

    2016-01-01

    One strategy for plants to optimize stomatal function is to open and close their stomata quickly in response to environmental signals. It is generally assumed that small stomata can alter aperture faster than large stomata. We tested the hypothesis that species with small stomata close faster than species with larger stomata in response to darkness by comparing rate of stomatal closure across an evolutionary range of species including ferns, cycads, conifers, and angiosperms under controlled ambient conditions (380 ppm CO2; 20.9% O2). The two species with fastest half-closure time and the two species with slowest half-closure time had large stomata while the remaining three species had small stomata, implying that closing rate was not correlated with stomatal size in these species. Neither was response time correlated with stomatal density, phylogeny, functional group, or life strategy. Our results suggest that past atmospheric CO2 concentration during time of taxa diversification may influence stomatal response time. We show that species which last diversified under low or declining atmospheric CO2 concentration close stomata faster than species that last diversified in a high CO2 world. Low atmospheric [CO2] during taxa diversification may have placed a selection pressure on plants to accelerate stomatal closing to maintain adequate internal CO2 and optimize water use efficiency. PMID:27605929

  2. Does size matter? Atmospheric CO2 may be a stronger driver of stomatal closing rate than stomatal size in taxa that diversified under low CO2.

    Directory of Open Access Journals (Sweden)

    Caroline Elliott-Kingston

    2016-08-01

    Full Text Available (1 One strategy for plants to optimise stomatal function is to open and close their stomata quickly in response to environmental signals. It is generally assumed that small stomata can alter aperture faster than large stomata. (2 We tested the hypothesis that species with small stomata close faster than species with larger stomata in response to darkness by comparing rate of stomatal closure across an evolutionary range of species including ferns, cycads, conifers and angiosperms under controlled ambient conditions (380ppm CO2; 20.9% O2. (3 The two species with fastest half-closure time and the two species with slowest half-closure time had large stomata while the remaining three species had small stomata, implying that closing rate was not correlated with stomatal size in these species. Neither was response time correlated with stomatal density, phylogeny, functional group or life strategy. (4 Our results suggest that past atmospheric CO2 concentration during time of taxa diversification may influence stomatal response time. We show that species which last diversified under low or declining atmospheric CO2 concentration close stomata faster than species that last diversified in a high CO2 world. Low atmospheric [CO2] during taxa diversification may have placed a selection pressure on plants to accelerate stomatal closing to maintain adequate internal CO2 and optimise water use efficiency.

  3. Stomatal function, density and pattern, and CO2 assimilation in Arabidopsis thaliana tmm1 and sdd1-1 mutants.

    Science.gov (United States)

    Vráblová, M; Vrábl, D; Hronková, M; Kubásek, J; Šantrůček, J

    2017-09-01

    Stomata modulate the exchange of water and CO2 between plant and atmosphere. Although stomatal density is known to affect CO2 diffusion into the leaf and thus photosynthetic rate, the effect of stomatal density and patterning on CO2 assimilation is not fully understood. We used wild types Col-0 and C24 and stomatal mutants sdd1-1 and tmm1 of Arabidopsis thaliana, differing in stomatal density and pattern, to study the effects of these variations on both stomatal and mesophyll conductance and CO2 assimilation rate. Anatomical parameters of stomata, leaf temperature and carbon isotope discrimination were also assessed. Our results indicate that increased stomatal density enhanced stomatal conductance in sdd1-1 plants, with no effect on photosynthesis, due to both unchanged photosynthetic capacity and decreased mesophyll conductance. Clustering (abnormal patterning formed by clusters of two or more stomata) and a highly unequal distribution of stomata between the adaxial and abaxial leaf sides in tmm1 mutants also had no effect on photosynthesis. Except at very high stomatal densities, stomatal conductance and water loss were proportional to stomatal density. Stomatal formation in clusters reduced stomatal dynamics and their operational range as well as the efficiency of CO2 transport. © 2017 German Botanical Society and The Royal Botanical Society of the Netherlands.

  4. AMLEXANOX 5% SEBAGAI MODALITAS TERAPI STOMATITIS AFTOSA REKUREN TERKINI

    Directory of Open Access Journals (Sweden)

    Rusmawati Rusmawati

    2015-07-01

    Full Text Available Recurrent Aphthous Stomatitis (RAS is characterized by painful recurring ulceration of oral mucosa and is the most common oral ulcerative disease. The specific etiology of the disease remains unknown. This situation makes the therapy of RAS difficult and uncertain. This article will discuss the effectiveness and the safety of amlexanox 5% in relieving the symptoms and accelerate heling in RAS lesions. Amlexanox 5% is a new drug used topically in treatment of RAS. The mechanism of action by which alexanox accelerates healing of RAS in unknown. Clnincal study of efficacy amlexanox 5% has been demonstrated significantly accelerate healing and reduce pain of RAS, compared to amlexanox 1% vehicle and no treatment. 5% amlexanox oral paste has an advantageous pharmacological activity to accelerate healing and reduce the pain of RAS, thus it can be used as one of the modality in this therapy.

  5. Allergic contact stomatitis caused by acrylic monomer in a denture.

    Science.gov (United States)

    Koutis, D; Freeman, S

    2001-08-01

    A 71-year-old edentulous man developed a severely painful red mouth at sites of contact with a new denture. Patch testing showed allergy to samples of the denture material and to 2-hydroxyethyl methacrylate. Patch testing to methyl methacrylate was negative. Prolonged boiling of the denture resulted in reversal of his symptoms and samples of this fully cured denture material produced negative patch tests. While allergy to acrylates is a rare cause of stomatitis, this possibility must be considered in patients presenting with oral symptoms. Material safety data sheets are unreliable in providing information regarding the type of acrylate present in the material. Hence, patch testing should be performed with a battery of acrylate allergens as well as with small samples of the denture material.

  6. Effect of bedtime on recurrent aphthous stomatitis in college students.

    Science.gov (United States)

    Ma, Ruiyang; Chen, Hong; Zhou, Tengfei; Chen, Xiyan; Wang, Chaoling; Chen, Yijin; Rao, Songlin; Ge, Lin; Lin, Mei

    2015-02-01

    In this study, we conducted a questionnaire-based survey at Sichuan University to investigate the potential effects of bedtime on recurrent aphthous stomatitis (RAS). An anonymous self-designed questionnaire was adopted to investigate the association between bedtime and RAS in college students at Sichuan University. Statistical analyses were used to identify risk factors for RAS and to explore the relationship between bedtime and RAS. One thousand six students were investigated. High frequency of colds (odds ratio [OR] 2.17; 95% confidence interval [CI] 1.52-3.10; P factors for RAS recurrence, but family history, stress, bad relationships with roommates, and gastrointestinal disease were not. Moreover, increased frequency of bedtime after 11 p.m. (τ > 0; P 0; P factor but both its frequency and cumulative time were also associated with severity of RAS in college students. Copyright © 2015 Elsevier Inc. All rights reserved.

  7. Stomatal design principles in synthetic and real leaves

    DEFF Research Database (Denmark)

    Zwieniecki, Maciej A.; Haaning, Katrine S; Boyce, C. Kevin

    2016-01-01

    Stomata are portals in plant leaves that control gas exchange for photosynthesis, a process fundamental to life on Earth. Gas fluxes and plant productivity depend on external factors such as light, water and CO2 availability and on the geometrical properties of the stoma pores. The link between...... stoma geometry and environmental factors has informed a wide range of scientific fields-from agriculture to climate science, where observed variations in stoma size and density are used to infer prehistoric atmospheric CO2 content. However, the physical mechanisms and design principles responsible...... efficient use of space and maximum control of dynamic gas conductivity, and that the capacity for gas exchange in plants has remained constant over at least the last 325 Myr. Our analysis provides a new measure to gauge the relative performance of species based on their stomatal characteristics....

  8. Nonredundant functions of Arabidopsis LecRK-V.2 and LecRK-VII.1 in controlling stomatal immunity and jasmonate-mediated stomatal closure.

    Science.gov (United States)

    Yekondi, Shweta; Liang, Fu-Chun; Okuma, Eiji; Radziejwoski, Amandine; Mai, Hsien-Wei; Swain, Swadhin; Singh, Prashant; Gauthier, Mathieu; Chien, Hsiao-Chiao; Murata, Yoshiyuki; Zimmerli, Laurent

    2017-12-18

    Stomatal immunity restricts bacterial entry to leaves through the recognition of microbe-associated molecular patterns (MAMPs) by pattern-recognition receptors (PRRs) and downstream abscisic acid and salicylic acid signaling. Through a reverse genetics approach, we characterized the function of the L-type lectin receptor kinase-V.2 (LecRK-V.2) and -VII.1 (LecRK-VII.1). Analyses of interactions with the PRR FLAGELLIN SENSING2 (FLS2) were performed by co-immunoprecipitation and bimolecular fluorescence complementation and whole-cell patch-clamp analyses were used to evaluate guard cell Ca2+ -permeable cation channels. The Arabidopsis thaliana LecRK-V.2 and LecRK-VII.1 and notably their kinase activities were required for full activation of stomatal immunity. Knockout lecrk-V.2 and lecrk-VII.1 mutants were hyper-susceptible to Pseudomonas syringae infection and showed defective stomatal closure in response to bacteria or to the MAMPs flagellin and EF-Tu. By contrast, Arabidopsis over-expressing LecRK-V.2 or LecRK-VII.1 demonstrated a potentiated stomatal immunity. LecRK-V.2 and LecRK-VII.1 are shown to be part of the FLS2 PRR complex. In addition, LecRK-V.2 and LecRK-VII.1 were critical for methyl jasmonate (MeJA)-mediated stomatal closure, notably for MeJA-induced activation of guard cell Ca2+ -permeable cation channels. This study highlights the role of LecRK-V.2 and LecRK-VII.1 in stomatal immunity at the FLS2 PRR complex and in MeJA-mediated stomatal closure. © 2017 The Authors. New Phytologist © 2017 New Phytologist Trust.

  9. Does smoking really protect from recurrent aphthous stomatitis?

    Directory of Open Access Journals (Sweden)

    Faleh A Sawair

    2010-11-01

    Full Text Available Faleh A SawairFaculty of Dentistry, University of Jordan, Amman, JordanPurpose: To study the effect of smoking on the prevalence of recurrent aphthous stomatitis (RAS and to examine whether intensity and duration of smoking influence RAS lesions.Subjects and methods: A cross-sectional survey was conducted on a random sample of 1000 students of The University of Jordan, Amman, between May and September 2008. Sociodemographic factors and details about smoking habits and RAS in last 12 months were collected.Results: Annual prevalence (AP of RAS was 37.1%. Tobacco use was common among students: 30.2% were current smokers and 2.8% were exsmokers. AP was not significantly influenced by students’ age, gender, marital status, college, and household income but was significantly affected by place of living (P = 0.02 and presence of chronic diseases (P = 0.03. No significant difference in AP of RAS was found between smokers and nonsmokers. Cigarette smokers who smoked heavily and for a longer period of time had significantly less AP of RAS when compared to moderate smokers and those who smoked for a shorter period of time. The protective effect of smoking was only noticed when there was heavy cigarette smoking (>20 cigarettes/day (P = 0.021 or smoking for long periods of time (>5 years (P = 0.009. Nevertheless, no significant associations were found between intensity or duration of smoking and clinical severity of RAS lesions.Conclusion: The “protective effect” of smoking on RAS was dose- and time-dependent. When lesions are present, smoking had no effect on RAS severity.Keywords: recurrent aphthous stomatitis, smoking, prevalence

  10. Genetic variation in circadian regulation of nocturnal stomatal conductance enhances carbon assimilation and growth.

    Science.gov (United States)

    Resco de Dios, Víctor; Loik, Michael E; Smith, Renee; Aspinwall, Michael J; Tissue, David T

    2016-01-01

    Circadian resonance, whereby a plant's endogenous rhythms are tuned to match environmental cues, has been repeatedly shown to be adaptive, although the underlying mechanisms remain elusive. Concomitantly, the adaptive value of nocturnal transpiration in C3 plants remains unknown because it occurs without carbon assimilation. These seemingly unrelated processes are interconnected because circadian regulation drives temporal patterns in nocturnal stomatal conductance, with maximum values occurring immediately before dawn for many species. We grew individuals of six Eucalyptus camaldulensis genotypes in naturally lit glasshouses and measured sunset, predawn and midday leaf gas exchange and whole-plant biomass production. We tested whether sunrise anticipation by the circadian clock and subsequent increases in genotype predawn stomatal conductance led to rapid stomatal opening upon illumination, ultimately affecting genotype differences in carbon assimilation and growth. We observed faster stomatal responses to light inputs at sunrise in genotypes with higher predawn stomatal conductance. Moreover, early morning and midday stomatal conductance and carbon assimilation, leaf area and total plant biomass were all positively correlated with predawn stomatal conductance across genotypes. Our results lead to the novel hypothesis that genotypic variation in the circadian-regulated capacity to anticipate sunrise could be an important factor underlying intraspecific variation in tree growth. © 2015 John Wiley & Sons Ltd.

  11. Natural variation in stomatal responses to environmental changes among Arabidopsis thaliana ecotypes.

    Directory of Open Access Journals (Sweden)

    Sho Takahashi

    Full Text Available Stomata are small pores surrounded by guard cells that regulate gas exchange between plants and the atmosphere. Guard cells integrate multiple environmental signals and control the aperture width to ensure appropriate stomatal function for plant survival. Leaf temperature can be used as an indirect indicator of stomatal conductance to environmental signals. In this study, leaf thermal imaging of 374 Arabidopsis ecotypes was performed to assess their stomatal responses to changes in environmental CO2 concentrations. We identified three ecotypes, Köln (Kl-4, Gabelstein (Ga-0, and Chisdra (Chi-1, that have particularly low responsiveness to changes in CO2 concentrations. We next investigated stomatal responses to other environmental signals in these selected ecotypes, with Col-0 as the reference. The stomatal responses to light were also reduced in the three selected ecotypes when compared with Col-0. In contrast, their stomatal responses to changes in humidity were similar to those of Col-0. Of note, the responses to abscisic acid, a plant hormone involved in the adaptation of plants to reduced water availability, were not entirely consistent with the responses to humidity. This study demonstrates that the stomatal responses to CO2 and light share closely associated signaling mechanisms that are not generally correlated with humidity signaling pathways in these ecotypes. The results might reflect differences between ecotypes in intrinsic response mechanisms to environmental signals.

  12. Changes in stomatal traits and the covariation with other leaf traits along an altitude transect

    Science.gov (United States)

    Wang, Ruili; Yu, Guirui; He, Nianpeng; Ge, Jianping; Wang, Qiufeng; Zhao, Ning; Xu, Zhiwei

    2014-05-01

    Stomatal traits and their responses to the external environment have been intensively studied for individual plant species. However, little is known about general stomatal patterns along environmental gradients in a broad, interspecific context or about the relationship between stomatal traits and other leaf traits. Here, we measured the stomatal and leaf traits, including stomatal density (SD), stomatal length (SL), specific leaf area (SLA), leaf area (LA), leaf thickness (LT) and nitrogen concentration (mass- and area- base, Nmass and Narea) of 158 plant species along an altitudinal gradient on Changbai Mountain, China. Our results revealed that SD decreased and SL increased significantly with altitude for tree species, although no clear elevational trends were observed in SD and SL across species (including tree, shrub, and herbaceous plants). Plant growth forms (PGFs) were the most important driver of variation in SD and SL, and the contributions of the mean annual temperature, precipitation and soil water content were weak. In addition, a covarying relationship between stomatal and other leaf traits was observed, although this relationship changed with elevation. These findings reflect that the adaptive strategies of plant ecophysiological traits may be complex for alpine environmental gradients, combining the short-term plasticity to environmental changes and long-term convergent evolution.

  13. Stomatal limitation to carbon gain in Paphiopedilum sp. (Orchidaceae) and its reversal by blue light

    Energy Technology Data Exchange (ETDEWEB)

    Zeiger, E.; Grivet, C.; Assmann, S.M.; Dietzer, G.F.; Hannegan, M.W.

    1985-02-01

    Leaves from Paphiopedilum sp. (Orchidaceae) having achlorophyllous stomata, show reduced levels of stomatal conductance when irradiated with red light, as compared with either the related, chlorophyllous genus Phragmipedium or with their response to blue light. These reduced levels of stomatal conductance, and the failure of isolated Paphiopedilum stomata to open under red irradiation indicates that the small stomatal response measured in the intact leaf under red light is indirect. The overall low levels of stomatal conductance observed in Paphiopedilum leaves under most growing conditions and their capacity to increase stomatal conductance in response to blue light suggested that growth and carbon gain in Paphiopedilum could be enhanced in a blue light-enriched environment. To test that hypothesis, plants of Paphiopedilum acmodontum were grown in controlled growth chambers under daylight fluorescent light, with or without blue light supplementation. Blue light enrichment resulted in significantly higher growth rates over a 3 to 4 week growing period, with all evidence indicating that the blue light effect was a stomatal response. Manipulations of stomatal properties aimed at long-term carbon gains could have agronomic applications.

  14. Stomatal Limitation to Carbon Gain in Paphiopedilum sp. (Orchidaceae) and Its Reversal by Blue Light.

    Science.gov (United States)

    Zeiger, E; Grivet, C; Assmann, S M; Deitzer, G F; Hannegan, M W

    1985-02-01

    Leaves from Paphiopedilum sp. (Orchidaceae) having achlorophyllous stomata, show reduced levels of stomatal conductance when irradiated with red light, as compared with either the related, chlorophyllous genus Phragmipedium or with their response to blue light. These reduced levels of stomatal conductance, and the failure of isolated Paphiopedilum stomata to open under red irradiation indicates that the small stomatal response measured in the intact leaf under red light is indirect.The overall low levels of stomatal conductance observed in Paphiopedilum leaves under most growing conditions and their capacity to increase stomatal conductance in response to blue light suggested that growth and carbon gain in Paphiopedilum could be enhanced in a blue light-enriched environment. To test that hypothesis, plants of Paphiopedilum acmodontum were grown in controlled growth chambers under daylight fluorescent light, with or without blue light supplementation. Total photosynthetic photon flux density was kept constant in both conditions. Blue light enrichment resulted in significantly higher growth rates-of up to 77%-over a 3 to 4 week growing period, with all evidence indicating that the blue light effect was a stomatal response. Manipulations of stomatal properties aimed at long-term carbon gains could have agronomic applications.

  15. Stomatal Limitation to Carbon Gain in Paphiopedilum sp. (Orchidaceae) and Its Reversal by Blue Light 1

    Science.gov (United States)

    Zeiger, Eduardo; Grivet, C.; Assmann, Sarah M.; Deitzer, Gerald F.; Hannegan, M. W.

    1985-01-01

    Leaves from Paphiopedilum sp. (Orchidaceae) having achlorophyllous stomata, show reduced levels of stomatal conductance when irradiated with red light, as compared with either the related, chlorophyllous genus Phragmipedium or with their response to blue light. These reduced levels of stomatal conductance, and the failure of isolated Paphiopedilum stomata to open under red irradiation indicates that the small stomatal response measured in the intact leaf under red light is indirect. The overall low levels of stomatal conductance observed in Paphiopedilum leaves under most growing conditions and their capacity to increase stomatal conductance in response to blue light suggested that growth and carbon gain in Paphiopedilum could be enhanced in a blue light-enriched environment. To test that hypothesis, plants of Paphiopedilum acmodontum were grown in controlled growth chambers under daylight fluorescent light, with or without blue light supplementation. Total photosynthetic photon flux density was kept constant in both conditions. Blue light enrichment resulted in significantly higher growth rates—of up to 77%—over a 3 to 4 week growing period, with all evidence indicating that the blue light effect was a stomatal response. Manipulations of stomatal properties aimed at long-term carbon gains could have agronomic applications. PMID:16664074

  16. A rate equation model of stomatal responses to vapour pressure deficit and drought

    Directory of Open Access Journals (Sweden)

    Shanahan ST

    2002-08-01

    Full Text Available Abstract Background Stomata respond to vapour pressure deficit (D – when D increases, stomata begin to close. Closure is the result of a decline in guard cell turgor, but the link between D and turgor is poorly understood. We describe a model for stomatal responses to increasing D based upon cellular water relations. The model also incorporates impacts of increasing levels of water stress upon stomatal responses to increasing D. Results The model successfully mimics the three phases of stomatal responses to D and also reproduces the impact of increasing plant water deficit upon stomatal responses to increasing D. As water stress developed, stomata regulated transpiration at ever decreasing values of D. Thus, stomatal sensitivity to D increased with increasing water stress. Predictions from the model concerning the impact of changes in cuticular transpiration upon stomatal responses to increasing D are shown to conform to experimental data. Sensitivity analyses of stomatal responses to various parameters of the model show that leaf thickness, the fraction of leaf volume that is air-space, and the fraction of mesophyll cell wall in contact with air have little impact upon behaviour of the model. In contrast, changes in cuticular conductance and membrane hydraulic conductivity have significant impacts upon model behaviour. Conclusion Cuticular transpiration is an important feature of stomatal responses to D and is the cause of the 3 phase response to D. Feed-forward behaviour of stomata does not explain stomatal responses to D as feedback, involving water loss from guard cells, can explain these responses.

  17. Ultrastructure and development of non-contiguous stomatal clusters and helicocytic patterning in Begonia.

    Science.gov (United States)

    Rudall, Paula J; Julier, Adele C M; Kidner, Catherine A

    2017-11-23

    Helicocytic stomata are characterized by an inward spiral of mesogenous cells surrounding a central stomatal pore. They represent a relatively rare feature that occurs in some drought-tolerant angiosperm species. In some Begonia species with thick leaves, the stomata are not only helicocytic but also clustered into groups that are spaced apart by at least one cell. This paper presents a detailed ontogenetic study of this characteristic non-contiguous stomatal patterning in a developmental and phylogenetic context. Light microscopy and both scanning and transmission electron microscopy were used to examine stomatal development in several species of Begonia. Published reports of stomatal development in Begonia and other angiosperms were reviewed to provide a comprehensive discussion of the evolution of stomatal patterning. Helicocytic stomata develop from meristemoids that undergo a series of oriented asymmetric divisions to produce a spiral of mesogene stomatal lineage ground cells (SLGCs) surrounding a stoma. A clear developmental similarity between anisocytic and helicocytic stomata is positively correlated with the number of iterations of amplifying divisions that result in SLGCs. Stomatal clusters develop from asymmetric divisions in neighbouring SLGCs. Within each cluster, non-contiguous spacing of meristemoids is maintained by asymmetric divisions oriented away from each developing meristemoid. Formation of non-contiguous stomatal clusters in Begonia relies on two primary developmental factors in the epidermis: an inwardly spiralling series of amplifying divisions that result in helicocytic stomata, and the development of a variable number of meristemoids from neighbouring SLGCs within each cluster. Optimization of these features on an angiosperm phylogeny indicates that the occurrence of amplifying divisions could be pre-adaptive for these factors. Both factors have been thoroughly studied in terms of developmental genetics in Arabidopsis, suggesting gene

  18. Depletion of vesicular zinc in dorsal horn of spinal cord causes increased neuropathic pain in mice

    DEFF Research Database (Denmark)

    Jo, Seung; Danscher, Gorm; Schrøder, Henrik

    2008-01-01

    pain. The animals were then sacrificed 5 days later. The ZnT3 immunoreactivity was found to have decreased significantly in dorsal horn of fourth, fifth, and sixth lumbar segments. In parallel with the depressed ZnT3 immunoreactivity the amount of vesicular zinc decreased perceptibly in superficial...... to neuropathic pain we applied Chung's rodent pain model on BALB/c mice, and traced zinc transporter 3 (ZnT3) proteins and zinc ions with immunohistochemistry and autometallography (AMG), respectively. Under anesthesia the left fifth lumbar spinal nerve was ligated in male mice in order to produced neuropathic...... gray matters of especially layer I-IV of the same segments. The transection-induced reduction of vesicular zinc in ZEN terminals of the dorsal horn was synchronic to reduced pain threshold, as measured by von Frey method. In a separate study, we observed intensive zinc selenite precipitation in somata...

  19. Exclusion of close linkage between the synaptic vesicular monoamine transporter locus and schizophrenia spectrum disorders

    Energy Technology Data Exchange (ETDEWEB)

    Persico, A.M.; Uhl, G.R. [Johns Hopkins Univ. School of Medicine, Baltimore, MD (United States); Wang, Zhe Wu [Universitario Campus Bio-Medico, Rome (Italy)] [and others

    1995-12-18

    The principal brain synaptic vesicular monoamine transporter (VMAT2) is responsible for the reuptake of serotonin, dopamine, norepinephrine, epinephrine, and histamine from the cytoplasm into synaptic vesicles, thus contributing to determination of the size of releasable neurotransmitter vesicular pools. Potential involvement of VMAT2 gene variants in the etiology of schizophrenia and related disorders was tested using polymorphic VMAT2 gene markers in 156 subjects from 16 multiplex pedigrees with schizophrenia, schizophreniform, schizoaffective, and schizotypal disorders and mood incongruent psychotic depression. Assuming genetic homogeneity, complete ({theta} = 0.0) linkage to the schizophrenia spectrum was excluded under both dominant and recessive models. Allelic variants at the VMAT2 locus do not appear to provide major genetic contributions to the etiology of schizophrenia spectrum disorders in these pedigrees. 16 refs.

  20. A comparison of two stomatal conductance models for ozone flux modelling using data from two Brassica species

    Energy Technology Data Exchange (ETDEWEB)

    Op de Beeck, M., E-mail: maarten.opdebeeck@ua.ac.b [Research Group of Plant and Vegetation Ecology, University of Antwerp, Campus Drie Eiken, Department of Biology, Universiteitsplein 1, 2160 Wilrijk (Belgium); De Bock, M., E-mail: maarten.debock@ua.ac.b [Research Group of Molecular Plant Physiology and Biotechnology, University of Antwerp, Campus Groenenborger, Department of Biology, Groenenborgerlaan 171, 2020 Antwerpen (Belgium); Vandermeiren, K., E-mail: kavan@var.fgov.b [Veterinary and Agrochemical Research Centre (VAR), Leuvensesteenweg 17, 3080 Tervuren (Belgium); Temmerman, L. de, E-mail: ludet@var.fgov.b [Veterinary and Agrochemical Research Centre (VAR), Leuvensesteenweg 17, 3080 Tervuren (Belgium); Ceulemans, R., E-mail: reinhart.ceulemans@ua.ac.b [Research Group of Plant and Vegetation Ecology, University of Antwerp, Campus Drie Eiken, Department of Biology, Universiteitsplein 1, 2160 Wilrijk (Belgium)

    2010-10-15

    In this study we tested and compared a multiplicative stomatal model and a coupled semi-empirical stomatal-photosynthesis model in their ability to predict stomatal conductance to ozone (g{sub st}) using leaf-level data from oilseed rape (Brassica napus L.) and broccoli (Brassica oleracea L. var. italica Plenck). For oilseed rape, the multiplicative model and the coupled model were able to explain 72% and 73% of the observed g{sub st} variance, respectively. For broccoli, the models were able to explain 53% and 51% of the observed g{sub st} variance, respectively. These results support the coupled semi-empirical stomatal-photosynthesis model as a valid alternative to the multiplicative stomatal model for O{sub 3} flux modelling, in terms of predictive performance. - A multiplicative stomatal model and a coupled semi-empirical stomatal-photosynthesis model performed equally well when tested against leaf-level data for oilseed rape and broccoli.

  1. Recurrent aphthous stomatitis: topical treatment with minocycline and other evidence-based agents.

    Science.gov (United States)

    Zur, Eyal

    2012-01-01

    Tetracycline is known as an effective agent in the treatment of recurrent aphthous stomatitis. A less-known treatment option for this pathology is minocycline. Many articles have been published in the International Journal of Pharmaceutical Compounding about recurrent aphthous stomatitis, however, this is the first article published in the journal that concentrates on the evidence-based data of minocycline as a promising agent to treat this pathology in what some scientific reports indicate to be a more efficacious manner than tetracycline. Few options to compound topical preparations based on minocycline and other evidence-based agents are proposed in the treatment of recurrent aphthous stomatitis.

  2. Vesicular noradrenaline stores in peripheral nerves of the rat and their modification by tranylcypromine.

    OpenAIRE

    Fillenz, M; Stanford, S. C.

    1981-01-01

    1 Vesicular noradrenaline stores were compared in the heart, salivary gland and vas deferens of the rat. 2 Noradrenaline storage vesicles in nerve terminals of different organs differed with respect to the amount of noradrenaline they contain in the endogenous store (content), the amount of exogenous noradrenaline they can take up from the circulation (uptake) and the amount of noradrenaline they contain when they are saturated (total storage capacity). 3 The data suggest that the vesicles in...

  3. Endoplasmosis and exoplasmosis: the evolutionary principles underlying endocytosis, exocytosis, and vesicular transport

    OpenAIRE

    Schmid, Johannes A.

    2016-01-01

    Summary Eukaryotic cells are characterized by a multicompartmental structure with a?variety of organelles. Vesicular transport between these compartments requires membrane fusion events. Based on a?membrane topology view, we conclude that there are two basic mechanisms of membrane fusion, namely where the membranes first come in contact with the cis-side (the plasmatic phase of the lipid bilayer) or with the trans-side (the extra-plasmatic face). We propose to designate trans-membrane fusion ...

  4. Efficacy study of vesicular gel containing methotrexate and menthol combination on parakeratotic rat skin model.

    Science.gov (United States)

    Nagle, Amrita; Goyal, Amit K; Kesarla, Rajesh; Murthy, Rayasa R

    2011-06-01

    Methotrexate (MTX) is indicated in the symptomatic control of severe, recalcitrant, and disabling psoriasis. The oral or parenteral route of administration causes systemic toxicity. The topical route of delivery, though, reduces systemic toxicity and has limited applicability due to restricted permeability. Liposomal and niosomal MTX topical formulations have also been investigated with limited success to achieve drug localization in the skin. Menthol has been suggested in conditions of psoriasis, in addition to its skin-penetration-enhancing effect on drugs. The present work aimed at investigating the potential benefits of combining menthol with MTX in a vesicular gel base for not only improving the penetration and dermal availability of MTX, but also to render such a formulation more effective with greater patient acceptability. MTX liposomes were prepared by thin-film hydration, and the vesicles were characterized for drug-entrapment efficiency, size, and morphology. These liposomal vesicles were incorporated in a gel base, and this vesicular gel was evaluated for transdermal drug permeation and extent of drug accumulation in the skin, using a rat skin ex vivo model. Skin histology studies were carried out to investigate any structural changes caused by the permeation enhancers. Antipsoriatic efficacy of the formulations was tested in vivo, using the rat tail model. The results indicated that the vesicular gel containing menthol could cause maximum drug retention in the skin. The skin treated with menthol had a disrupted epidermis and microcavities. The in vivo studies also ascertained the effectiveness of the formulation in inducing a normal pattern of differentiation in the rat tail skin that initially showed parakeratosis, which is also characteristic of psoriatic epidermis. These results show the potential of vesicular gel containing MTX and menthol to improve penetration into the skin and cause drug retention in skin appendages.

  5. Formation and Stability of Prebiotically Relevant Vesicular Systems in Terrestrial Geothermal Environments

    OpenAIRE

    Manesh Prakash Joshi; Anupam Samanta; Gyana Ranjan Tripathy; Sudha Rajamani

    2017-01-01

    Terrestrial geothermal fields and oceanic hydrothermal vents are considered as candidate environments for the emergence of life on Earth. Nevertheless, the ionic strength and salinity of oceans present serious limitations for the self-assembly of amphiphiles, a process that is fundamental for the formation of first protocells. Consequently, we systematically characterized the efficiency of amphiphile assembly, and vesicular stability, in terrestrial geothermal environments, both, under simula...

  6. Combining sap flow and eddy covariance approaches to derive stomatal and non-stomatal O3 fluxes in a forest stand.

    Science.gov (United States)

    Nunn, A J; Cieslik, S; Metzger, U; Wieser, G; Matyssek, R

    2010-06-01

    Stomatal O3 fluxes to a mixed beech/spruce stand (Fagus sylvatica/Picea abies) in Central Europe were determined using two different approaches. The sap flow technique yielded the tree-level transpiration, whereas the eddy covariance method provided the stand-level evapotranspiration. Both data were then converted into stomatal ozone fluxes, exemplifying this novel concept for July 2007. Sap flow-based stomatal O3 flux was 33% of the total O3 flux, whereas derivation from evapotranspiration rates in combination with the Penman-Monteith algorithm amounted to 47%. In addition to this proportional difference, the sap flow-based assessment yielded lower levels of stomatal O3 flux and reflected stomatal regulation rather than O3 exposure, paralleling the daily courses of canopy conductance for water vapor and eddy covariance-based total stand-level O3 flux. The demonstrated combination of sap flow and eddy covariance approaches supports the development of O3 risk assessment in forests from O3 exposure towards flux-based concepts. Copyright 2009 Elsevier Ltd. All rights reserved.

  7. Consistent allometric scaling of stomatal sizes and densities across taxonomic ranks and geologic time

    Science.gov (United States)

    de Boer, H. J.; Price, C. A.; Wagner-Cremer, F.; Dekker, S. C.; Veneklaas, E. J.

    2013-12-01

    Stomatal pores on plants leaves are an important link in the chain of processes that determine biosphere fluxes of water and carbon. Stomatal density (i.e. the number of stomata per area) and the size of the stomatal pore at maximum aperture are particularly relevant traits in this context because they determine the theoretical maximum diffusive stomatal conductance (gsmax) and thereby set an upper limit for leaf gas exchange. Observations on (sub)fossil leaves revealed that changes in stomatal densities are anti-correlated with changes in stomatal sizes at developmental and evolutionary timescales. Moreover, this anti-correlation appears consistently within single species, across multiple species in the extant plant community and at evolutionary time scales. The consistency of the relation between stomatal densities and sizes suggests that common mechanisms constrain the adaptation of these traits across the plant community. In an attempt to identify such potential generic constraints, we investigated the allometry between stomatal densities and sizes in the extant plant community and across geological time. As the size of the stomatal pore at maximum aperture is typically derived from the length of the stomatal pore, we considered the allometric scaling of pore length (lp) with stomatal density (Ds) as the power law: lp = k . Dsa in which k is a normalization constant and the exponent a is the slope of the scaling relation. Our null-hypothesis predicts that stomatal density and pore length scale along a constant slope of -1/2 based on a scale-invariant relation between pore length and the distance between neighboring pores. Our alternative hypothesis predicts a constant slope of -1 based on the idea that stomatal density and pore length scale along an invariant gsmax. To explore these scaling hypotheses in the extant plant community we compiled a dataset of combined observations of stomatal density and pore length on 111 species from published literature and new

  8. Design, synthesis and biological evaluation of small-azo-dyes as potent Vesicular Glutamate Transporters inhibitors.

    Science.gov (United States)

    Favre-Besse, Franck-Cyril; Poirel, Odile; Bersot, Tiphaine; Kim-Grellier, Elodie; Daumas, Stephanie; El Mestikawy, Salah; Acher, Francine C; Pietrancosta, Nicolas

    2014-05-06

    Vesicular Glutamate Transporters (VGLUTs) allow the loading of presynapic glutamate vesicles and thus play a critical role in glutamatergic synaptic transmission. VGLUTs have proved to be involved in several major neuropathologies and directly correlated to clinical dementia in Alzheimer and Parkinson's disease. Accordingly VGLUT represent a key biological target or biomarker for neuropathology treatment or diagnostic. Yet, despite the pivotal role of VGLUTs, their pharmacology appears quite limited. Known competitive inhibitors are restricted to some dyes as Trypan Blue (TB) and glutamate mimics. This lack of pharmacological tools has heavily hampered VGLUT investigations. Here we report a rapid access to small molecules that combine benefits of TB and dicarboxylic quinolines (DCQs). Their ability to block vesicular glutamate uptake was evaluated. Several compounds displayed low micromolar inhibitory potency when size related compounds are thirty to forty times less potent (i.e. DCQ). We then confirmed the VGLUT selectivity by measuring the effect of the series on vesicular monoamine transport and on metabotropic glutamate receptor activity. These inhibitors are synthesized in only two steps and count among the best pharmacological tools for VGLUTs studies. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

  9. Inhibition of skin inflammation in mice by diclofenac in vesicular carriers: liposomes, ethosomes and PEVs.

    Science.gov (United States)

    Caddeo, Carla; Sales, Octavio Diez; Valenti, Donatella; Saurí, Amparo Ruiz; Fadda, Anna Maria; Manconi, Maria

    2013-02-25

    Diclofenac-loaded phospholipid vesicles, namely conventional liposomes, ethosomes and PEVs (penetration enhancer-containing vesicles) were developed and their efficacy in TPA (phorbol ester) induced skin inflammation was examined. Vesicles were made from a cheap and unpurified mixture of phospholipids and diclofenac sodium; Transcutol P and propylene glycol were added to obtain PEVs, and ethanol to produce ethosomes. The structure and lamellar organization of the vesicle bilayer were investigated by transmission electron microscopy and small and wide angle X-ray scattering, as well as the main physico-chemical features. The formulations, along with a diclofenac solution and commercial Voltaren Emulgel, were tested in a comparative trial for anti-inflammatory efficacy on TPA-treated mice dorsal skin. Vesicles were around 100 nm, negatively charged, able to encapsulate diclofenac in good yields, and disclosed different lamellarity, as a function of the formulation composition. Vesicular formulations promoted drug accumulation and reduced the permeation. Administration of vesicular diclofenac on TPA-inflamed skin resulted in marked attenuation of oedema and leucocyte infiltration, especially using PEVs. Histology confirmed the effectiveness of vesicles, since they provided an amelioration of the tissual damage induced by TPA. The proposed approach based on vesicular nanocarriers may hold promising therapeutic value for treating a variety of inflammatory skin disorders. Copyright © 2013 Elsevier B.V. All rights reserved.

  10. In vitro percutaneous permeation and skin accumulation of finasteride using vesicular ethosomal carriers.

    Science.gov (United States)

    Rao, Yuefeng; Zheng, Feiyue; Zhang, Xingguo; Gao, Jianqing; Liang, Wenquan

    2008-01-01

    In order to develop a novel transdermal drug delivery system that facilitates the skin permeation of finasteride encapsulated in novel lipid-based vesicular carriers (ethosomes)finasteride ethosomes were constructed and the morphological characteristics were studied by transmission electron microscopy. The particle size, zeta potential and the entrapment capacity of ethosome were also determined. In contrast to liposomes ethosomes were of more condensed vesicular structure and they were found to be oppositely charged. Ethosomes were found to be more efficient delivery carriers with high encapsulation capacities. In vitro percutaneous permeation experiments demonstrated that the permeation of finasteride through human cadaver skin was significantly increased when ethosomes were used. The finasteride transdermal fluxes from ethosomes containing formulation (1.34 +/- 0.11 microg/cm(2)/h) were 7.4, 3.2 and 2.6 times higher than that of finasteride from aqueous solution, conventional liposomes and hydroethanolic solution respectively (P ethosomes produced a significant (P ethosomes are promising vesicular carriers for enhancing percutaneous absorption of finasteride.

  11. Spiroindolines identify the vesicular acetylcholine transporter as a novel target for insecticide action.

    Directory of Open Access Journals (Sweden)

    Ann Sluder

    Full Text Available The efficacy of all major insecticide classes continues to be eroded by the development of resistance mediated, in part, by selection of alleles encoding insecticide insensitive target proteins. The discovery of new insecticide classes acting at novel protein binding sites is therefore important for the continued protection of the food supply from insect predators, and of human and animal health from insect borne disease. Here we describe a novel class of insecticides (Spiroindolines encompassing molecules that combine excellent activity against major agricultural pest species with low mammalian toxicity. We confidently assign the vesicular acetylcholine transporter as the molecular target of Spiroindolines through the combination of molecular genetics in model organisms with a pharmacological approach in insect tissues. The vesicular acetylcholine transporter can now be added to the list of validated insecticide targets in the acetylcholine signalling pathway and we anticipate that this will lead to the discovery of novel molecules useful in sustaining agriculture. In addition to their potential as insecticides and nematocides, Spiroindolines represent the only other class of chemical ligands for the vesicular acetylcholine transporter since those based on the discovery of vesamicol over 40 years ago, and as such, have potential to provide more selective tools for PET imaging in the diagnosis of neurodegenerative disease. They also provide novel biochemical tools for studies of the function of this protein family.

  12. Influence of the preparation route on the supramolecular organization of lipids in a vesicular system.

    Science.gov (United States)

    Elizondo, Elisa; Larsen, Jannik; Hatzakis, Nikos S; Cabrera, Ingrid; Bjørnholm, Thomas; Veciana, Jaume; Stamou, Dimitrios; Ventosa, Nora

    2012-02-01

    A confocal fluorescence microscopy-based assay was used for studying the influence of the preparation route on the supramolecular organization of lipids in a vesicular system. In this work, vesicles composed of cholesterol and CTAB (1/1 mol %) or cholesterol and DOPC (2/8 mol %) and incorporating two membrane dyes were prepared by either a compressed fluid (CF)-based method (DELOS-susp) or a conventional film hydration procedure. They were subsequently immobilized and imaged individually using a confocal fluorescence microscope. Two integrated fluorescence intensities, I(dye1) and I(dye2), were assigned to each tracked vesicle, and their ratio, I(dye1)/I(dye2), was used for quantifying the degree of membrane inhomogeneity between individual vesicles within each sample. A distribution of I(dye1)/I(dye2) values was obtained for all the studied vesicular systems, indicating intrasample heterogeneity. The degree of inhomogeneity (DI) was similar for Chol/DOPC vesicles prepared by both procedures. In contrast, DI was more than double for the hydration method compared to the CF-based method in the case of Chol/CTAB vesicles, which can suffer from lipid demixing during film formation. These findings reveal a more homogeneous vesicle formation path by CFs, which warranted good homogeneity of the vesicular system, independently of the lipid mixture used. © 2011 American Chemical Society

  13. Effect of two percent turmeric extract gel on minor recurrent aphthous stomatitis

    National Research Council Canada - National Science Library

    Nurdiana Nurdiana; Shamini Krishnasamy

    2017-01-01

    Minor recurrent aphthous stomatitis (RAS) is recurrent oral ulcer with clinical features of shallow ulcer, round or oval shape, measuring less than 10 mm, covered with yellowish white pseudomembrane and surrounded by erythematous halo...

  14. Evaluation of the therapeutic effects of Aloe vera gel on minor recurrent aphthous stomatitis

    Directory of Open Access Journals (Sweden)

    Neda Babaee

    2012-01-01

    Conclusion: It seems likely that A.V. 2% oral gel is not only effective in decreasing the recurrent aphthous stomatitis patients′ pain score and wound size but also decreases the aphthous wound healing period.

  15. Periodic fever accompanied by aphthous stomatitis, pharyngitis and cervical adenitis syndrome (PFAPA syndrome) in adults

    National Research Council Canada - National Science Library

    Padeh, Shai; Stoffman, Nava; Berkun, Yackov

    2008-01-01

    The new syndrome, known as PFAPA, of periodic fever characterized by abrupt onset of fever, malaise, aphthous stomatitis, tonsillitis, pharyngitis and cervical adenopathy has been described only in pediatric patients...

  16. Serum cytokine profile and clinicopathological findings in oral lichen planus, oral lichenoid lesions and stomatitis

    DEFF Research Database (Denmark)

    Larsen, Kristine Røn; Johansen, Jeanne Duus; Reibel, Jesper

    2017-01-01

    The objective of this study was to examine if clinical and histopathological variables in patients with oral lichen planus (OLP), oral lichenoid lesions (OLL), and generalized stomatitis display different cytokine profiles and if concomitant contact allergy influences this profile. Forty...

  17. The effect of dietary habits on the development of the recurrent aphthous stomatitis

    OpenAIRE

    Bassel Tarakji; Kusai Baroudi; Yaser Kharma

    2012-01-01

    Background: The aim was to assess the relationship between the dietary habits and development of recurrent aphthous stomatitis. Materials and Methods: Two groups (30 patients with RAS who have been following dietary habits and not associated with systemic disease or hematologic abnormalities, and the control group consist of 28 patients without recurrent aphthous stomatitis). Results: A Mann-Whitney test (P>0.05) shows no significance difference between the patients with RAS and the control g...

  18. Les caractéristiques des stomates des feuilles de Ficus benjamina L ...

    African Journals Online (AJOL)

    SARAH

    25 mars 2014 ... mm² d'épiderme (Hetherington & Woodward, 2003). La densité des stomates augmente avec l'intensité de la pollution de l'air en indiquant les zones les plus polluées et les zones les moins polluées (Woodward et al., 1995). La résistance des stomates (RS) exprime la mesure de l'inhibition de la diffusion ...

  19. Protective effect of new hygienic means on the oral mucous tunic at radial stomatitis

    OpenAIRE

    Ієвлєва, Юлія Валеріївна

    2015-01-01

    The radial stomatitis development is the one of the main problems in patients who underwent radial therapy of the malignant tumors in head and neck region that impedes the effective antitumor treatment. The means of prophylaxis and local treatment of radial injuries are not always effective.Aim of research. The determination of effectiveness of the new hygienic means based on antimatters on the state of oral mucous tunic at the radial stomatitis.Materials and methods of research. Experiments ...

  20. Stomatal VPD response: there is more to the story than ABA.

    Science.gov (United States)

    Merilo, Ebe; Yarmolinsky, Dmitry; Jalakas, Pirko; Parik, Helen; Tulva, Ingmar; Rasulov, Bakhtier; Kilk, Kalle; Kollist, Hannes

    2017-10-06

    Guard cells shrink and close stomatal pores when air humidity decreases, i.e. when the difference between the vapor pressures of leaf and atmosphere (VPD) increases. The role of abscisic acid (ABA) in VPD-induced stomatal closure has been studied using ABA-related mutants that respond to VPD in some studies and not in others. The importance of ABA biosynthesis in guard cells versus vasculature for the whole-plant stomatal regulation is also unclear. Here we show that Arabidopsis lines carrying mutations in different steps of ABA biosynthesis as well as pea wilty and tomato flacca ABA-deficient mutants had higher stomatal conductance compared to wildtype plants. To characterize the role of ABA production in different cells, we generated transgenic plants where ABA biosynthesis was rescued in guard cells or phloem companion cells of an ABA-deficient mutant. In both cases, the whole-plant stomatal conductance, stunted growth phenotype and leaf ABA level were restored to wildtype values, pointing to the redundancy of ABA sources and to the effectiveness of leaf ABA transport. All ABA-deficient lines closed their stomata rapidly and extensively in response to high VPD, whereas plants with mutated protein kinase OST1 showed stunted VPD-induced responses. Another strongly ABA-insensitive mutant defective in the six ABA PYR/RCAR receptors, responded to changes in VPD in both directions strongly and symmetrically, indicating that its VPD-induced closure could be passive hydraulic. We discuss that both the VPD-induced passive hydraulic stomatal closure and the stomatal VPD-regulation of ABA-deficient mutants may be conditional on the initial pretreatment stomatal conductance. {copyright, serif} 2017 American Society of Plant Biologists. All rights reserved.

  1. Level of Denture Cleanliness Influences the Presence of Denture Stomatitis on Maxillary Denture Bearing-Mucosa

    OpenAIRE

    Winatty Krisma; Martha Mozartha; Rani Purba

    2014-01-01

    Plaque accumulation on internal surface of denture is a common problem among removable denture wearers. Poor denture cleanliness can increase colonization of Candida albicans and cause inflammatory reaction of denture-bearing mucosa, i.e. denture stomatitis. Objective: To find out the effect of denture cleanliness level on denture stomatitis on maxillary denture-bearing mucosa in a group of removable denture wearers who received prosthodontic treatment at Poliklinik Gigi RSMH Palembang and to...

  2. Anamnestic findings from patients with recurrent aphthous stomatitis.

    Science.gov (United States)

    Bratel, John; Hakeberg, Magnus

    2014-01-01

    Recurrent aphthous stomatitis (RAS) is a common oral disorder with a prevalence varying between 5% and 66%. RAS appears in three forms; minor, major and herpetiform. The aetiology is unknown.The aim of this study was to evaluate associations between specific anamnestic information and different types of recurrent aphthous stomatitis (RAS). A group of 177 patients (mean age = 42.8 years; SD = 14.3; range 17-79 years) participated. Data were collected from a structured interview, consisting of 22 questions. Information about i) health status and medication, ii) predisposing factors, iii) RAS experience, iv) previous treatment methods and v) brand of toothpaste was collected. Sixty-eight per cent of the patients were healthy and 44% of the patients were not taking any medication. Forty-one per cent of the patients did not have any apprehension of the reason for their RAS, while stress (15.8%) was the most common apprehended aetiological factor. Sixty-two per cent had one to three minor ulcers at one time. Forty-eight per cent reported having had a major aphthous ulcer at least once.The most frequent symptom reported was pain (53.7%), followed by a smarting sensation (18.6%) and tenderness (4%). The most common treatment for RAS was Zendium™ toothpaste/mouthrinse (28%), followed by corticosteroids (25%). Fifty-four per cent of the patients experienced no relief from the treatment. When toothpaste habits were investigated, Zendium™ was used by 32% of the patients and toothpaste containing sodium-lauryl-sulfatase was used by 32%.There was no positive correlation between the use of Zendium™ toothpaste and the relief of symptoms or the size, number or frequency of the aphthous ulcers. Sixty-four per cent of the patients had never smoked, while 7% were smokers. No positive correlation was found when age, gender, allergy, medication and smoking were correlated to the frequency, number and size of the aphthous ulcers. In conclusion, we found that the aetiology behind

  3. Predicting photosynthesis and transpiration responses to ozone: decoupling modeled photosynthesis and stomatal conductance

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    D. Lombardozzi

    2012-08-01

    Full Text Available Plants exchange greenhouse gases carbon dioxide and water with the atmosphere through the processes of photosynthesis and transpiration, making them essential in climate regulation. Carbon dioxide and water exchange are typically coupled through the control of stomatal conductance, and the parameterization in many models often predict conductance based on photosynthesis values. Some environmental conditions, like exposure to high ozone (O3 concentrations, alter photosynthesis independent of stomatal conductance, so models that couple these processes cannot accurately predict both. The goals of this study were to test direct and indirect photosynthesis and stomatal conductance modifications based on O3 damage to tulip poplar (Liriodendron tulipifera in a coupled Farquhar/Ball-Berry model. The same modifications were then tested in the Community Land Model (CLM to determine the impacts on gross primary productivity (GPP and transpiration at a constant O3 concentration of 100 parts per billion (ppb. Modifying the Vcmax parameter and directly modifying stomatal conductance best predicts photosynthesis and stomatal conductance responses to chronic O3 over a range of environmental conditions. On a global scale, directly modifying conductance reduces the effect of O3 on both transpiration and GPP compared to indirectly modifying conductance, particularly in the tropics. The results of this study suggest that independently modifying stomatal conductance can improve the ability of models to predict hydrologic cycling, and therefore improve future climate predictions.

  4. Quantification of stomatal uptake of ionic solutes using a new model system.

    Science.gov (United States)

    Eichert, T; Burkhardt, J

    2001-04-01

    Evidence for stomatal uptake of solutes by leaves without the application of surfactants or pressure has recently been provided (Eichert et al., 1998). In the present study, experimental conditions were refined in that the water potential was held at Allium porrum, Commelina communis and Sedum telephium. Uptake increased with humidity, stomatal aperture and stomatal density. It was restricted to stomatal areas, and was especially high below the rims of drying droplets. Again, penetration of stomatal pores was observed. Uptake was strongly correlated with the number of penetrated stomata, although usually less than 10% of the stomata contributed to uptake. The number of stomata that had been penetrated was highly variable, increasing extremely significantly with the number of repeated drying/ wetting cycles. These results indicate that stomatal uptake can be a major pathway for the foliar uptake of ionic solutes. It is a dynamic process, depending on environmental conditions and history of the residues on the leaf, aspects that had been neglected in previous concepts.

  5. Expression of Arabidopsis hexokinase in citrus guard cells controls stomatal aperture and reduces transpiration

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    Nitsan eLugassi

    2015-12-01

    Full Text Available Hexokinase (HXK is a sugar-phosphorylating enzyme involved in sugar-sensing. It has recently been shown that HXK in guard cells mediates stomatal closure and coordinates photosynthesis with transpiration in the annual species tomato and Arabidopsis. To examine the role of HXK in the control of the stomatal movement of perennial plants, we generated citrus plants that express Arabidopsis HXK1 (AtHXK1 under KST1, a guard cell-specific promoter. The expression of KST1 in the guard cells of citrus plants has been verified using GFP as a reporter gene. The expression of AtHXK1 in the guard cells of citrus reduced stomatal conductance and transpiration with no negative effect on the rate of photosynthesis, leading to increased water-use efficiency. The effects of light intensity and humidity on stomatal behavior were examined in rooted leaves of the citrus plants. The optimal intensity of photosynthetically active radiation and lower humidity enhanced stomatal closure of AtHXK1-expressing leaves, supporting the role of sugar in the regulation of citrus stomata. These results suggest that HXK coordinates photosynthesis and transpiration and stimulates stomatal closure not only in annual species, but also in perennial species.

  6. Salicaceae Endophytes Modulate Stomatal Behavior and Increase Water Use Efficiency in Rice

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    Hyungmin Rho

    2018-03-01

    Full Text Available Bacterial and yeast endophytes isolated from the Salicaceae family have been shown to promote growth and alleviate stress in plants from different taxa. To determine the physiological pathways through which endophytes affect plant water relations, we investigated leaf water potential, whole-plant water use, and stomatal responses of rice plants to Salicaceae endophyte inoculation under CO2 enrichment and water deficit. Daytime stomatal conductance and stomatal density were lower in inoculated plants compared to controls. Leaf ABA concentrations increased with endophyte inoculation. As a result, transpirational water use decreased significantly with endophyte inoculation while biomass did not change or slightly increased. This response led to a significant increase in cumulative water use efficiency at harvest. Different endophyte strains produced the same results in host plant water relations and stomatal responses. These stomatal responses were also observed under elevated CO2 conditions, and the increase in water use efficiency was more pronounced under water deficit conditions. The effect on water use efficiency was positively correlated with daily light integrals across different experiments. Our results provide insights on the physiological mechanisms of plant-endophyte interactions involving plant water relations and stomatal functions.

  7. Putting the brakes on: abscisic acid as a central environmental regulator of stomatal development.

    Science.gov (United States)

    Chater, Caspar C C; Oliver, James; Casson, Stuart; Gray, Julie E

    2014-04-01

    Stomata are produced by a controlled series of epidermal cell divisions. The molecular underpinnings of this process are becoming well understood, but mechanisms that determine plasticity of stomatal patterning to many exogenous and environmental cues remain less clear. Light quantity and quality, vapour pressure deficit, soil water content, and CO2 concentration are detected by the plant, and new leaves adapt their stomatal densities accordingly. Mature leaves detect these environmental signals and relay messages to immature leaves to tell them how to adapt and grow. Stomata on mature leaves may act as stress signal-sensing and transduction centres, locally by aperture adjustment, and at long distance by optimizing stomatal density to maximize future carbon gain while minimizing water loss. Although mechanisms of stomatal aperture responses are well characterized, the pathways by which mature stomata integrate environmental signals to control immature epidermal cell fate, and ultimately stomatal density, are not. Here we evaluate current understanding of the latter through the influence of the former. We argue that mature stomata, as key portals by which plants coordinate their carbon and water relations, are controlled by abscisic acid (ABA), both metabolically and hydraulically, and that ABA is also a core regulator of environmentally determined stomatal development. © 2014 The Authors. New Phytologist © 2014 New Phytologist Trust.

  8. Brazilian Green Propolis Compared to Miconazole Gel in the Treatment of Candida-Associated Denture Stomatitis

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    Hermínia Marques Capistrano

    2013-01-01

    Full Text Available Aim. To evaluate the efficacy of Brazilian green propolis in comparison to miconazole gel in the treatment of Candida-associated denture stomatitis. Methods. Forty-five denture stomatitis patients, with palatal mucosa erythema levels classified according to Newtons’s criteria and with positive culture to Candida spp., were randomly divided into three treatment groups: 15 received miconazole gel 2%, 15 received propolis gel 2,5%, and 15 received propolis 24% for mouthwash. After four daily use lasting two weeks, they were reexamined for the denture stomatitis degree and for a second culture of Candida. The Wilcoxon’s test was applied to compare the results of clinical classification of the denture stomatitis and the Candida spp. colonies numbers, before and after each treatment. The Kruskall-Wallis’s test was used to compare efficacy among the three treatment groups. Results. There were a significant reduction or complete remission of denture stomatitis (P0.05. Conclusion. Brazilian green propolis has a similar effect as miconazole in the treatment of Candida-associated denture stomatitis being an alternative in the therapeutics of this condition.

  9. Level of Denture Cleanliness Influences the Presence of Denture Stomatitis on Maxillary Denture Bearing-Mucosa

    Directory of Open Access Journals (Sweden)

    Winatty Krisma

    2014-12-01

    Full Text Available Plaque accumulation on internal surface of denture is a common problem among removable denture wearers. Poor denture cleanliness can increase colonization of Candida albicans and cause inflammatory reaction of denture-bearing mucosa, i.e. denture stomatitis. Objective: To find out the effect of denture cleanliness level on denture stomatitis on maxillary denture-bearing mucosa in a group of removable denture wearers who received prosthodontic treatment at Poliklinik Gigi RSMH Palembang and to investigate the denture hygiene habits of removable denture wearers. Methods: Thirty subjects participated in this study. Denture cleanliness level was assessed with disclosing solution to disclose denture plaque on internal surface of maxillary denture. Cleanliness level was graded according to Budtz-Jorgensen. Intraoral examination was done to determine any visible signs of denture stomatitis. Data referring to denture hygiene habits of removable denture wearers was collected from interview using questionnaire. Data were analyzed using the Komolgorov-Smirnov test. Results: Result of the study showed that 40% subjects had poor upper denture cleanliness. Denture stomatitis was observed on maxillary denture-bearing mucosa in 43.3% subjects. Kolmogorov-Smirnov test showed that there was a significant effect of denture cleanliness level on denture stomatitis on maxillary denture-bearing mucosa (p<0.05. Conclusion: Denture cleanliness level influence the occurence of denture stomatitis on maxillary denture bearing-mucosa in a group of removable denture wearers who received prosthodontic treatment.

  10. The effect of ascorbate on minor recurrent aphthous stomatitis.

    Science.gov (United States)

    Yasui, K; Kurata, T; Yashiro, M; Tsuge, M; Ohtsuki, S; Morishima, T

    2010-03-01

    Minor recurrent aphthous stomatitis (MRAS) is a common, painful and inflammatory ailment of the oral cavity with juvenile onset and unknown aetiology. The purpose of this study was to evaluate the potential of ascorbate (vitamin C) to reduce the frequency of MRAS and severity of pain. Sixteen MRAS patients (9 boys and 7 girls: mean age, 12.0 +/- 2.4 years old) were assigned to take an oral dosage of 2000 mg/m(2)/day ascorbate. Their baseline frequency of outbreaks and the level of pains were compared during the treatment; in addition, a crossover clinical trial was performed. Polymorphonuclear leucocytes play a role in the pathogenesis, and then superoxide anion production was evaluated in prior to ascorbate treatment. The data indicated a statistically significant 50% reduction in oral ulcer outbreaks and a decline of pain level. Neutrophils were primed for superoxide anion production in the patients with MRAS. Ascorbate may modulate the generation of reactive oxygen species and augment neutrophil apoptosis, which could prevent neutrophil-mediated inflammation. Ascorbate seems to be effective, but the findings of our study were preliminary and it should be re-evaluated with a larger randomized controlled clinical trials.

  11. Management strategies for HIV-associated aphthous stomatitis.

    Science.gov (United States)

    Kerr, A Ross; Ship, Jonathan A

    2003-01-01

    Recurrent aphthous stomatitis (RAS) is the most common oral mucosal disorder found in men and women of all ages, races, and geographic regions. There are three forms of the lesions (minor, major, and herpetiform), with major aphthous ulcers causing significant pain and potential for scarring. In HIV-infected individuals, these ulcers occur more frequently, last longer, and produce more painful symptoms than in immunocompetent persons. In addition, they may be associated with similar ulcerations involving the esophagus, rectum, anus, and genitals. The diagnosis of HIV-induced RAS requires a careful history of the condition, and a thorough extra- and intra-oral examination. Oral mucosal biopsies are required for non-healing ulcers in order to exclude the possibility of deep fungal infections, viral infections, and neoplasms. The cause of the ulcers in HIV-positive persons has not been elucidated--local diseases, genetic, immunologic, and infectious factors all probably play a role. The goals of current treatments are to promote ulcer healing, to reduce ulcer duration and pain while maintaining nutritional intake, and to prevent or diminish the frequency of recurrence. Initial therapy for infrequent RAS recurrences includes over-the-counter topical protective and analgesic products. Initial therapy for frequent RAS outbreaks requires topical anesthetics, binding agents, and corticosteroids. Major RAS and non-healing minor or herpetiform RAS may require intralesional corticosteroids and systemic prednisone. Second-line immunomodulators for frequent and non-healing ulcers includes thalidomide and other immunomodulators.

  12. The oral microbiota of patients with recurrent aphthous stomatitis

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    Maria Bankvall

    2014-10-01

    Full Text Available Background: Specific pathogenic bacteria have been implicated in recurrent aphthous stomatitis (RAS, a chronic inflammatory condition characterised by ulcerations in the oral mucosa. However, the aetiology behind this condition still remains unclear. Objective: The buccal microbiota of patients with RAS was compared to that of control subjects to investigate its potential role for this condition. Design: Buccal swabs were obtained from non-ulcerative areas of 60 patients, of whom 42 patients had lesions at the time of sampling, and 60 healthy age- and gender-matched controls. Bacterial DNA was extracted and analysed by Terminal-Restriction Fragment Length Polymorphism, using enzymatic digestion of the polymerase chain reaction-amplified 16S rRNA gene, yielding a series of peaks, each representing a bacterial taxon. Results: Two peaks, 60 and 489, were more prevalent in patients with RAS than controls. Conversely, peaks 58 and 490 were less common in patients than controls. When the patients were divided into subgroups, we found that the observed differences in peak-pattern were related to the presence of lesions during sampling. Conclusions: The microbiota of the non-inflamed buccal mucosa differed between patients and controls. The differences were most pronounced in patients who presented with lesions during sampling, suggesting that a disturbance in the normal buccal microbiota triggers the presence of lesions or that presence of lesions alters the microbiota.

  13. Subclinical atherosclerosis in patients with recurrent aphthous stomatitis.

    Science.gov (United States)

    Sereflican, M; Sereflican, B; Dagistan, E; Goksugur, N; Kizildag, B

    2016-09-01

    Recurrent aphtous stomatitis (RAS) is an inflammatory oral mucosal disease. It has been known that inflammatory cascade plays important role in the atherosclerotic process. The aim of the study was to investigate the relationship between subclinical atherosclerotic findings and a systemic inflammatory disease, RAS. In total, 32 patients with RAS were matched with 30 control subjects on the basis of age, sex, and major cardiovascular risk factors. Laboratory parameters including lipid profiles were determined for patients and controls. B-mode ultrasonography was used to assess carotid extra-medial thickness (cEMT) and carotid intima-media thickness (cIMT). Both cEMT and cIMT in the RAS group were significantly higher than in the control group (P = 0.002 and 0.013, respectively). There was a significant positive correlation between cIMT and cEMT (r = 0.381, P = 0.034). cIMT was positively correlated with age, triglyceride levels, and systolic blood pressure, while cEMT was positively correlated with age in patients with RAS. To our knowledge, this is the first reported study to evaluate cEMT and cIMT in patients with RAS. This study presents morphological evidence of subclinical atherosclerosis in patients with RAS. Further studies investigating the relationship between atherosclerosis and RAS are needed. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  14. The oral microbiota of patients with recurrent aphthous stomatitis

    Science.gov (United States)

    Bankvall, Maria; Sjöberg, Fei; Gale, Gita; Wold, Agnes; Jontell, Mats; Östman, Sofia

    2014-01-01

    Background Specific pathogenic bacteria have been implicated in recurrent aphthous stomatitis (RAS), a chronic inflammatory condition characterised by ulcerations in the oral mucosa. However, the aetiology behind this condition still remains unclear. Objective The buccal microbiota of patients with RAS was compared to that of control subjects to investigate its potential role for this condition. Design Buccal swabs were obtained from non-ulcerative areas of 60 patients, of whom 42 patients had lesions at the time of sampling, and 60 healthy age- and gender-matched controls. Bacterial DNA was extracted and analysed by Terminal-Restriction Fragment Length Polymorphism, using enzymatic digestion of the polymerase chain reaction-amplified 16S rRNA gene, yielding a series of peaks, each representing a bacterial taxon. Results Two peaks, 60 and 489, were more prevalent in patients with RAS than controls. Conversely, peaks 58 and 490 were less common in patients than controls. When the patients were divided into subgroups, we found that the observed differences in peak-pattern were related to the presence of lesions during sampling. Conclusions The microbiota of the non-inflamed buccal mucosa differed between patients and controls. The differences were most pronounced in patients who presented with lesions during sampling, suggesting that a disturbance in the normal buccal microbiota triggers the presence of lesions or that presence of lesions alters the microbiota. PMID:25626771

  15. Treatment of recurrent aphthous stomatitis major with metronidazole and ciprofloxacin

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    M. Jusri

    2009-09-01

    Full Text Available Background: Recurrent aphthous stomatitis (RAS are painful oral ulcerations that recur from days to months or even years. It represents the most common lesion of the oral mucosa with prevalence ranging from 15% to 30%. Although the clinical characteristics of RAS are well defined, the precise etiopathogenesis of RAS remains unclear. Since the etiology of RAS remains unknown, there is no definitive treatment. RAS responds quite well to the use of topical or systemic antiinflammatory drugs, particularly corticosteroids. Purpose: The objective of this paper is to discuss the treatment of RAS with secondary infection. Case: This paper reported a case of 22-year-old man with multiple oral ulcers that did not heal for 7 months. Case Management: These ulcers were diagnosed as RAS major with secondary infection that caused by normal oral flora (aerobic and anaerobic bacteria and treated with metronidazole (topical and oral and ciprofloxacin (oral. These lesions healed in 3 weeks with scars. Conclusion: Large ulcer without signs of malignancy that contaminated with normal oral flora will delayed in healing, but with rational treatment RAS mayor with secondary infection has good prognosis.

  16. Efficacy of alum for treatment of recurrent aphthous stomatitis.

    Science.gov (United States)

    Rafieian, Nasrin; Abdolsamadi, Hamidreza; Moghadamnia, Aliakbar; Jazayeri, Mina; Seif-Rabiee, Mohammadali; Salmanzadeh, Mina; Radi, Shahrbanoo

    2016-01-01

    Recurrent aphthous stomatitis (RAS) is the most common painful ulcers of oral mucosal which can cause many sufferings. Treatment of RAS often includes administration of corticosteroids, analgesics and regulators of the immune system. However, considering the side effects of these medications, even their topical application must be done with caution. Alum is used in traditional medicine for treatment of oral ulcers without significant side effect. This study sought to assess the effect of topical application of alum on aphthous ulcers. This clinical randomized double-blind placebo-controlled study was conducted on 50 females aged 21 to 27 years. Mucosal adhesive patches were prepared in two forms of basic and 7% alum-containing patches. Subjects in two groups of case and control randomly received the mucosal adhesive patches containing alum and the basic patches, respectively three times in five days. Duration of recovery, changes in size of lesion and severity of pain were recorded. Data were entered into SPSS Version 16 and analyzed using t-test. The average period of full recovery was 7.52 days in the case and 12.2 days in the control groups; which was significantly different (paphthous lesions, severity of pain and expedite the recovery of patients with RAS.

  17. Validation of Anamnestic Diagnostic Criteria for Recurrent Aphthous Stomatitis

    Science.gov (United States)

    Baccaglini, Lorena; Theriaque, Douglas W.; Shuster, Jonathan J.; Serrano, Giselle; Lalla, Rajesh V.

    2014-01-01

    BACKGROUND Recurrent aphthous stomatitis (RAS) is characterized by painful recurrent oral ulcers and is typically diagnosed via history and clinical examination. Our aim was to validate a set of anamnestic diagnostic criteria (RASDX) to increase the accuracy of RAS diagnosis, particularly when a clinical examination is not feasible. METHODS Participants were enrolled during an unmatched case-control study. RASDX consisted of an initial phone screening using standardized questionnaires and recognition of RAS photos in the clinic. The proportion of agreement with an examination by an oral medicine expert was calculated. RESULTS A total of 115 participants were scheduled for a clinical diagnostic visit and 11 were withdrawn. The remaining 104 participants were aged 18–50 years, 54% female, 64% White and 20% Hispanic. Of these, all 49 controls with negative RASDX had no clinical ulcers. Of the 54 cases diagnosed with RAS by RASDX, 53 were clinically confirmed to have RAS lesions (99% agreement; exact 1-sided 95% CI=95–100%). CONCLUSIONS RASDX, based on a combination of history and photograph recognition, was highly accurate compared to a diagnosis that employed an oral examination. PMID:23106421

  18. A review on the oxidative stress in recurrent aphtous stomatitis

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    Mehryari Mahsa

    2015-01-01

    Full Text Available   Background and Aims: The purpose of the present study was to review the studies regarding serological and salivary oxidant / antioxidant status in patients with recurrent aphthous stomatitis (RAS.   Materials and Methods: The literature was searched using key words RAS, Antioxidants, Saliva, Hematinic (s and Hematinic Acid in the last 10 years, particularly the recent 3 years (2010-2013. At total of 37 clinical trials, 18 case-control articles were selected and evaluated; fulfilling the requirements as the RAS patients having at least 3 oral aphthous attack per year. The exclusion criteria included systemic as well as periodontal diseases, iron deficiency associated anemia, medication usage and smoking.   Conclusion: Almost all lipid-peroxidation studies in serum and saliva were manifested by an increase of malondialdehyde (MDA concentration in RAS patients compared with controls. This would indicate the role of reactive oxygen species (ROS in the etiology of the disease. Serum trace elements (Zn, Se were reduced and Cu was increased in RAS patients in comparison with control individuals. A decreased serum superoxide dismutase (SOD and an increased salivary SOD were observed in all RAS patients. Catalase (CAT and uric acid (UA analyses were non-inclusive. Levels of paraoxonase and arylesterase as well as antioxidant vitamins (A, E, C were lower in RAS patients than that of controls.

  19. Ozone treatment of recurrent aphthous stomatitis: a double blinded study.

    Science.gov (United States)

    Al-Omiri, Mahmoud K; Alhijawi, Mohannad; AlZarea, Bader K; Abul Hassan, Ra'ed S; Lynch, Edward

    2016-06-15

    This study aimed to evaluate the use of ozone to treat recurrent aphthous stomatitis (RAS). Consecutive sixty-nine participants with RAS were recruited into this non-randomized double blind, controlled cohort observational study (test group). A control group of 69 RAS patients who matched test group with age and gender was recruited. RAS lesions in test group were exposed to ozone in air for 60 seconds while controls received only air. Ulcer size and pain were recorded for each participant at baseline and daily for 15 days. Ulcer duration was determined by recording the time taken for ulcers to disappear. The main outcome measures were pain due to the ulcer, ulcer size and ulcer duration. 138 RAS participants (69 participants and 69 controls) were analyzed. Ulcer size was reduced starting from the second day in test group and from the fourth day in controls (p ≤ 0.004). Pain levels were reduced starting from the first day in the test group and from the third day in controls (p ≤ 0.001). Ulcer duration, ulcer size after day 2 and pain levels were more reduced in the test group. In conclusion, application of ozone on RAS lesions for 60 seconds reduced pain levels and enhanced ulcers' healing by reducing ulcers' size and duration.

  20. A photosynthesis-based two-leaf canopy stomatal ...

    Science.gov (United States)

    A coupled photosynthesis-stomatal conductance model with single-layer sunlit and shaded leaf canopy scaling is implemented and evaluated in a diagnostic box model with the Pleim-Xiu land surface model (PX LSM) and ozone deposition model components taken directly from the meteorology and air quality modeling system—WRF/CMAQ (Weather Research and Forecast model and Community Multiscale Air Quality model). The photosynthesis-based model for PX LSM (PX PSN) is evaluated at a FLUXNET site for implementation against different parameterizations and the current PX LSM approach with a simple Jarvis function (PX Jarvis). Latent heat flux (LH) from PX PSN is further evaluated at five FLUXNET sites with different vegetation types and landscape characteristics. Simulated ozone deposition and flux from PX PSN are evaluated at one of the sites with ozone flux measurements. Overall, the PX PSN simulates LH as well as the PX Jarvis approach. The PX PSN, however, shows distinct advantages over the PX Jarvis approach for grassland that likely result from its treatment of C3 and C4 plants for CO2 assimilation. Simulations using Moderate Resolution Imaging Spectroradiometer (MODIS) leaf area index (LAI) rather than LAI measured at each site assess how the model would perform with grid averaged data used in WRF/CMAQ. MODIS LAI estimates degrade model performance at all sites but one site having exceptionally old and tall trees. Ozone deposition velocity and ozone flux along with LH

  1. Modeling of stomatal conductance to estimate stomatal ozone uptake by Fagus crenata, Quercus serrata, Quercus mongolica var. crispula and Betula platyphylla.

    Science.gov (United States)

    Kinose, Yoshiyuki; Azuchi, Fumika; Uehara, Yui; Kanomata, Tomoaki; Kobayashi, Ayumi; Yamaguchi, Masahiro; Izuta, Takeshi

    2014-11-01

    To construct stomatal conductance models and estimate stomatal O3 uptake for Fagus crenata, Quercus serrata, Quercus mongolica var. crispula and Betula platyphylla, stomatal conductance (gs) was measured in seedlings of the four tree species. Better estimates of gs were made by incorporating the acute effects of O3 on gs into the models and the models could explain 34-52% of the variability in gs. Although the O3 concentration was relatively high in spring from April to May, COU of F. crenata, Q. serrata and Q. mongolica var. crispula were relatively low and the ratios of COU in spring to total COU in one year were 16.8% in all tree species because of low gs limited mainly by leaf pre-maturation and/or low temperature. The COU of B. platyphylla were relatively high mainly because of rapid leaf maturation and lower optimal temperature for stomatal opening. Copyright © 2014 Elsevier Ltd. All rights reserved.

  2. Diagnosis and management of recurrent herpetiform stomatitis and Behçet syndrome like recurrent aphthous stomatitis herpetiform type

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    Endah Ayu Tri Wulandari

    2008-11-01

    Full Text Available Recurrent Aphthous Stomatitis (RAS is a common inflammatory condition of the oral mucosa. The aetiology of RAS remains unclear, yet there are several predisposing factors which could be involved in the onset of the lesion. The herpetiform type of RAS appeared to be similar to recurrent oral Herpes Simplex infection and also could be part of Behçet Syndrome. This case report discussed a patient suffering from a herpetiform type of RAS with its clinical appearance resembling recurrent oral Herpes Simplex infection and Behçet syndrome. Initial treatment was undertaken based on the empirical treatment, yet the respond was not satisfactory. Then, laboratory tests were undertaken, including complete blood count, the total population of T lymphocyte, B lymphocyte, T helper, T suppressor, NK cells, T helper/T suppressor ratio, C3, C4, IgG, IgA, and IgM. These tests showed that there were immune and hematinic deficiency condition. Nevertheless, the clinical appearance, laboratory findings and consultation did not support the diagnosis of recurrent oral Herpes Simplex infection and Behçet Syndrome, thus, enhancing the definite diagnosis of the herpetiform type of RAS with immune and hematinic deficiency as the underlying condition. Based on the definite diagnosis, treatment plan was then revised to target the underlying condition.

  3. Coxiella burnetii effector protein subverts clathrin-mediated vesicular trafficking for pathogen vacuole biogenesis

    Science.gov (United States)

    Larson, Charles L.; Beare, Paul A.; Howe, Dale; Heinzen, Robert A.

    2013-01-01

    Successful macrophage colonization by Coxiella burnetii, the cause of human Q fever, requires pathogen-directed biogenesis of a large, growth-permissive parasitophorous vacuole (PV) with phagolysosomal characteristics. The vesicular trafficking pathways co-opted by C. burnetii for PV development are poorly defined; however, it is predicted that effector proteins delivered to the cytosol by a defective in organelle trafficking/intracellular multiplication (Dot/Icm) type 4B secretion system are required for membrane recruitment. Here, we describe involvement of clathrin-mediated vesicular trafficking in PV generation and the engagement of this pathway by the C. burnetii type 4B secretion system substrate Coxiella vacuolar protein A (CvpA). CvpA contains multiple dileucine [DERQ]XXXL[LI] and tyrosine (YXXΦ)-based endocytic sorting motifs like those recognized by the clathrin adaptor protein (AP) complexes AP1, AP2, and AP3. A C. burnetii ΔcvpA mutant exhibited significant defects in replication and PV development, confirming the importance of CvpA in infection. Ectopically expressed mCherry-CvpA localized to tubular and vesicular domains of pericentrosomal recycling endosomes positive for Rab11 and transferrin receptor, and CvpA membrane interactions were lost upon mutation of endocytic sorting motifs. Consistent with CvpA engagement of the endocytic recycling system, ectopic expression reduced uptake of transferrin. In pull-down assays, peptides containing CvpA-sorting motifs and full-length CvpA interacted with AP2 subunits and clathrin heavy chain. Furthermore, depletion of AP2 or clathrin by siRNA treatment significantly inhibited C. burnetii replication. Thus, our results reveal the importance of clathrin-coated vesicle trafficking in C. burnetii infection and define a role for CvpA in subverting these transport mechanisms. PMID:24248335

  4. Scaling of stomatal size and density optimizes allocation of leaf epidermal space for gas exchange in angiosperms

    Science.gov (United States)

    de Boer, Hugo Jan; Price, Charles A.; Wagner-Cremer, Friederike; Dekker, Stefan C.; Franks, Peter J.; Veneklaas, Erik J.

    2015-04-01

    Stomata on plant leaves are key traits in the regulation of terrestrial fluxes of water and carbon. The basic morphology of stomata consists of a diffusion pore and two guard cells that regulate the exchange of CO2 and water vapour between the leaf interior and the atmosphere. This morphology is common to nearly all land plants, yet stomatal size (defined as the area of the guard cell pair) and stomatal density (the number of stomata per unit area) range over three orders of magnitude across species. Evolution of stomatal sizes and densities is driven by selection pressure on the anatomical maximum stomatal conductance (gsmax), which determines the operational range of leaf gas exchange. Despite the importance of stomata traits for regulating leaf gas exchange, a quantitative understanding of the relation between adaptation of gsmax and the underlying co-evolution of stomatal sizes and densities is still lacking. Here we develop a theoretical framework for a scaling relationship between stomatal sizes and densities within the constraints set by the allocation of epidermal space and stomatal gas exchange. Our theory predicts an optimal scaling relationship that maximizes gsmax and minimizes epidermal space allocation to stomata. We test whether stomatal sizes and densities reflect this optimal scaling with a global compilation of stomatal trait data on 923 species reflecting most major clades. Our results show optimal scaling between stomatal sizes and densities across all species in the compiled data set. Our results also show optimal stomatal scaling across angiosperm species, but not across gymnosperm and fern species. We propose that the evolutionary flexibility of angiosperms to adjust stomatal sizes underlies their optimal allocation of leaf epidermal space to gas exchange.

  5. A putative vesicular transporter expressed in Drosophila mushroom bodies that mediates sexual behavior may define a neurotransmitter system.

    Science.gov (United States)

    Brooks, Elizabeth S; Greer, Christina L; Romero-Calderón, Rafael; Serway, Christine N; Grygoruk, Anna; Haimovitz, Jasmine M; Nguyen, Bac T; Najibi, Rod; Tabone, Christopher J; de Belle, J Steven; Krantz, David E

    2011-10-20

    Vesicular transporters are required for the storage of all classical and amino acid neurotransmitters in synaptic vesicles. Some neurons lack known vesicular transporters, suggesting additional neurotransmitter systems remain unidentified. Insect mushroom bodies (MBs) are critical for several behaviors, including learning, but the neurotransmitters released by the intrinsic Kenyon cells (KCs) remain unknown. Likewise, KCs do not express a known vesicular transporter. We report the identification of a novel Drosophila gene portabella (prt) that is structurally similar to known vesicular transporters. Both larval and adult brains express PRT in the KCs of the MBs. Additional PRT cells project to the central complex and optic ganglia. prt mutation causes an olfactory learning deficit and an unusual defect in the male's position during copulation that is rescued by expression in KCs. Because prt is expressed in neurons that lack other known vesicular transporters or neurotransmitters, it may define a previously unknown neurotransmitter system responsible for sexual behavior and a component of olfactory learning. Copyright © 2011 Elsevier Inc. All rights reserved.

  6. ANTICUERPOS CONTRA EL VIRUS DE ESTOMATITIS VESICULAR EN HUANGANAS (Tayassu pecari) EN MADRE DE DIOS, PERÚ

    OpenAIRE

    Carruitero H., Susan; Laboratorio de Microbiología y Parasitología Veterinaria, Facultad de Medicina Veterinaria, Universidad Nacional Mayor de San Marcos, Lima; Rivera G., Hermelinda; Laboratorio de Microbiología y Parasitología Veterinaria, Facultad de Medicina Veterinaria, Universidad Nacional Mayor de San Marcos, Lima Perú.; Ramírez V., Mercy; Laboratorio de Microbiología y Parasitología Veterinaria, Facultad de Medicina Veterinaria, Universidad Nacional Mayor de San Marcos, Lima; More B., Juan; Laboratorio de Microbiología y Parasitología Veterinaria, Facultad de Medicina Veterinaria, Universidad Nacional Mayor de San Marcos, Lima.; Zúñiga H., Alfonso; Proyecto ÁREAS–Amazonía de la World Wildlife Fund (WWF-Peru); Romero S., Mónica; Proyecto ÁREAS–Amazonía de la World Wildlife Fund (WWF-Peru)

    2013-01-01

    El objetivo del presente estudio fue determinar la presencia de anticuerpos neutralizantes contra los serotipos New Jersey (NJ) e Indiana subtipo 1 (IND-1) del virus Estomatitis Vesicular (VEV) en huanganas (Tayassu pecari) de vida libre de las localidades de Boca de Manu (n=30), Concesión para la Conservación Los Amigos (n=10) y La Reserva Nacional Tambopata/Parque Nacional Bahuaja Sonene (n=48) en el departamento de Madre de Dios. La presencia de anticuerpos contra el VEV fue determinado me...

  7. MEJORAMIENTO DE LA PRODUCCIÓN DE UNA VACUNA OLEOSA CONTRA ESTOMATITIS VESICULAR BIVALENTE

    OpenAIRE

    Arbeláez, Gustavo; Departamento de Microbiología, Facultad de Ciencias, Pontificia Universidad Javeriana, Cra. 7 Nº 43 - 82, Bogotá; Mondragón, Nestor; Empresa Colombiana de Productos Veterinarios Vecol S.A., Bogotá; Turriago, Clara; Empresa Colombiana de Productos Veterinarios Vecol S.A., Bogotá; Mora, Nelson; Empresa Colombiana de Productos Veterinarios Vecol S.A., Bogotá; Méndez, María; Empresa Colombiana de Productos Veterinarios Vecol S.A., Bogotá

    2008-01-01

    El presente estudio calculó diferentes MI (Multiplicidad de Infección) para la producción de cultivos industriales de virus de Estomatitis Vesicular (EV) y evaluó el efecto de la cantidad de glicoproteína G en la inducción de respuesta de anticuerpos neutralizantes contra el virus de EV en cobayos inmunizados con una vacuna oleosa bivalente (Indiana (I) y New Jersey (NJ)). Al establecer el MI más eficiente se logró mejorar la cinética de infección de los cultivos industriales disminuyendo los...

  8. The Effects of Vesicular-Arbuscular Mycorrhizae on the Plant Growth and Nutrient Uptake of Cucumber

    OpenAIRE

    ÇIĞŞAR, Sibel; Sari, Nebahat

    2014-01-01

    This study was conducted to investigate the effect of vesicular-arbuscular (VA) mycorrhizae on plant growth of cucumber. Yayla F 1 seeds were sown in sterile and non-sterile growing medium (organic manure:soil:mix of sand; v:v:v 1:1:1). The mix inoculum of Glomus mosseaand Glomus fasciculatumspores (10 g/plant) was placed 5 cm below the cucumber seed before sowing. In order to investigate the effects of VA mycorrhizae on plant growth, plant height, diameter, number of nodes were measured ...

  9. Stomatal cell wall composition: distinctive structural patterns associated with different phylogenetic groups

    Science.gov (United States)

    Shtein, Ilana; Shelef, Yaniv; Marom, Ziv; Zelinger, Einat; Schwartz, Amnon; Popper, Zoë A.; Bar-On, Benny

    2017-01-01

    Background and Aims Stomatal morphology and function have remained largely conserved throughout ∼400 million years of plant evolution. However, plant cell wall composition has evolved and changed. Here stomatal cell wall composition was investigated in different vascular plant groups in attempt to understand their possible effect on stomatal function. Methods A renewed look at stomatal cell walls was attempted utilizing digitalized polar microscopy, confocal microscopy, histology and a numerical finite-elements simulation. The six species of vascular plants chosen for this study cover a broad structural, ecophysiological and evolutionary spectrum: ferns (Asplenium nidus and Platycerium bifurcatum) and angiosperms (Arabidopsis thaliana and Commelina erecta) with kidney-shaped stomata, and grasses (angiosperms, family Poaceae) with dumbbell-shaped stomata (Sorghum bicolor and Triticum aestivum). Key Results Three distinct patterns of cellulose crystallinity in stomatal cell walls were observed: Type I (kidney-shaped stomata, ferns), Type II (kidney-shaped stomata, angiosperms) and Type III (dumbbell-shaped stomata, grasses). The different stomatal cell wall attributes investigated (cellulose crystallinity, pectins, lignin, phenolics) exhibited taxon-specific patterns, with reciprocal substitution of structural elements in the end-walls of kidney-shaped stomata. According to a numerical bio-mechanical model, the end walls of kidney-shaped stomata develop the highest stresses during opening. Conclusions The data presented demonstrate for the first time the existence of distinct spatial patterns of varying cellulose crystallinity in guard cell walls. It is also highly intriguing that in angiosperms crystalline cellulose appears to have replaced lignin that occurs in the stomatal end-walls of ferns serving a similar wall strengthening function. Such taxon-specific spatial patterns of cell wall components could imply different biomechanical functions, which in turn

  10. Stomatal cell wall composition: distinctive structural patterns associated with different phylogenetic groups.

    Science.gov (United States)

    Shtein, Ilana; Shelef, Yaniv; Marom, Ziv; Zelinger, Einat; Schwartz, Amnon; Popper, Zoë A; Bar-On, Benny; Harpaz-Saad, Smadar

    2017-04-01

    Stomatal morphology and function have remained largely conserved throughout ∼400 million years of plant evolution. However, plant cell wall composition has evolved and changed. Here stomatal cell wall composition was investigated in different vascular plant groups in attempt to understand their possible effect on stomatal function. A renewed look at stomatal cell walls was attempted utilizing digitalized polar microscopy, confocal microscopy, histology and a numerical finite-elements simulation. The six species of vascular plants chosen for this study cover a broad structural, ecophysiological and evolutionary spectrum: ferns ( Asplenium nidus and Platycerium bifurcatum ) and angiosperms ( Arabidopsis thaliana and Commelina erecta ) with kidney-shaped stomata, and grasses (angiosperms, family Poaceae) with dumbbell-shaped stomata ( Sorghum bicolor and Triticum aestivum ). Three distinct patterns of cellulose crystallinity in stomatal cell walls were observed: Type I (kidney-shaped stomata, ferns), Type II (kidney-shaped stomata, angiosperms) and Type III (dumbbell-shaped stomata, grasses). The different stomatal cell wall attributes investigated (cellulose crystallinity, pectins, lignin, phenolics) exhibited taxon-specific patterns, with reciprocal substitution of structural elements in the end-walls of kidney-shaped stomata. According to a numerical bio-mechanical model, the end walls of kidney-shaped stomata develop the highest stresses during opening. The data presented demonstrate for the first time the existence of distinct spatial patterns of varying cellulose crystallinity in guard cell walls. It is also highly intriguing that in angiosperms crystalline cellulose appears to have replaced lignin that occurs in the stomatal end-walls of ferns serving a similar wall strengthening function. Such taxon-specific spatial patterns of cell wall components could imply different biomechanical functions, which in turn could be a consequence of differences in

  11. Auxin represses stomatal development in dark-grown seedlings via Aux/IAA proteins.

    Science.gov (United States)

    Balcerowicz, Martin; Ranjan, Aashish; Rupprecht, Laura; Fiene, Gabriele; Hoecker, Ute

    2014-08-01

    Stomatal development is tightly regulated through internal and external factors that are integrated by a complex signalling network. Light represents an external factor that strongly promotes stomata formation. Here, we show that auxin-resistant aux/iaa mutants, e.g. axr3-1, exhibit a de-repression of stomata differentiation in dark-grown seedlings. The higher stomatal index in dark-grown axr3-1 mutants when compared with the wild type is due to increased cell division in the stomatal lineage. Excessive stomata in dark-grown seedlings were also observed in mutants defective in auxin biosynthesis or auxin perception and in seedlings treated with the polar auxin transport inhibitor NPA. Consistent with these findings, exogenous auxin repressed stomata formation in light-grown seedlings. Taken together, these results indicate that auxin is a negative regulator of stomatal development in dark-grown seedlings. Epistasis analysis revealed that axr3-1 acts genetically upstream of the bHLH transcription factors SPCH, MUTE and FAMA, as well as the YDA MAP kinase cascade, but in parallel with the repressor of photomorphogenesis COP1 and the receptor-like protein TMM. The effect of exogenous auxin required the ER family of leucine-rich repeat receptor-like kinases, suggesting that auxin acts at least in part through the ER family. Expression of axr3-1 in the stomatal lineage was insufficient to alter the stomatal index, implying that cell-cell communication is necessary to mediate the effect of auxin. In summary, our results show that auxin signalling contributes to the suppression of stomatal differentiation observed in dark-grown seedlings. © 2014. Published by The Company of Biologists Ltd.

  12. Clinical study on thermography, as modern investigation method for Candida-associated denture stomatitis.

    Science.gov (United States)

    Iosif, Laura; Preoteasa, Cristina Teodora; Murariu-Măgureanu, Cătălina; Preoteasa, Elena

    2016-01-01

    Candida-associated denture stomatitis is an infectious inflammatory condition of the oral mucosa, with frequent recurrences. The aim of this study was to assess the use of infrared thermography as investigation method for Candida-associated denture stomatitis (as inflammatory disorder of the maxillary denture bearing area), by comparing disease and non-disease groups. An observational study was conducted on maxillary edentulous patients treated by acrylic dentures, with and without Candida-associated denture stomatitis. Diagnostic test methods used were clinical examination for denture stomatitis and conventional microbiological culture method for oral candidiasis. Thermography analysis of the maxillary denture bearing area was made using the ThermaCAM PM350 infrared camera (Inframetrics, Flir Systems) and ThermaGram Pro 95 software, data being acquired by usage of standard protocol of thermographic registrations. The sample included 52 patients, 21 with and 31 without Candida-associated denture stomatitis. The temperature of the maxillary mucosa corresponding to the denture bearing area was found to be statistically significantly higher in Candida-associated denture stomatitis (mean 36.20°C), compared to healthy oral mucosa (mean 34.85°C). The thermal threshold value of 35.44°C was identified as best differentiating a pathological from normal state of the maxillary mucosa corresponding to the denture bearing area. In conclusion, infrared thermography, a rapid non-invasive investigation method, has the premises to bring valuable data in inflammatory disorders of the maxillary denture bearing area, as Candida-associated denture stomatitis that may be used for screening, diagnostic or monitoring purposes.

  13. Contrasting responses of leaf stomatal characteristics to climate change: a considerable challenge to predict carbon and water cycles.

    Science.gov (United States)

    Yan, Weiming; Zhong, Yangquanwei; Shangguan, Zhouping

    2017-09-01

    Stomata control the cycling of water and carbon between plants and the atmosphere; however, no consistent conclusions have been drawn regarding the response of stomatal frequency to climate change. Here, we conducted a meta-analysis of 1854 globally obtained data series to determine the response of stomatal frequency to climate change, which including four plant life forms (over 900 species), at altitudes ranging from 0 to 4500 m and over a time span of more than one hundred thousand years. Stomatal frequency decreased with increasing CO2 concentration and increased with elevated temperature and drought stress; it was also dependent on the species and experimental conditions. The response of stomatal frequency to climate change showed a trade-off between stomatal control strategies and environmental factors, such as the CO2 concentration, temperature, and soil water availability. Moreover, threshold effects of elevated CO2 and temperature on stomatal frequency were detected, indicating that the response of stomatal density to increasing CO2 concentration will decrease over the next few years. The results also suggested that the stomatal index may be more reliable than stomatal density for determination of the historic CO2 concentration. Our findings indicate that the contrasting responses of stomata to climate change bring a considerable challenge in predicting future water and carbon cycles. © 2017 John Wiley & Sons Ltd.

  14. Development of Candida-associated denture stomatitis: new insights

    Directory of Open Access Journals (Sweden)

    Tatiana Pereira-Cenci

    2008-04-01

    Full Text Available Despite therapeutic progress, opportunistic oral fungal infectious diseases have increased in prevalence, especially in denture wearers. The combination of entrapment of yeast cells in irregularities in denture-base and denture-relining materials, poor oral hygiene and several systemic factors is the most probable cause for the onset of this infectious disease. Hence colonization and growth on prostheses by Candida species are of clinical importance. The purpose of this review is to critically discuss several key factors controlling the adhesion of Candida species which are relevant to denture-associated stomatitis. Although there is some consensus on the role of surface properties, studies on several other factors, as the use of denture liners, salivary properties and yeast-bacterial interactions, have shown contradictory findings. A comprehensive fundamental understanding is hampered by conflicting findings due to the large variations in experimental protocols, while other factors have never been thoroughly studied. Surface free energy and surface roughness control the initial adherence, but temporal changes have not been reported. Neither have in vivo studies shown if the substratum type is critical in dictating biofilm accumulation during longer periods in the oral environment. The contribution of saliva is unclear due to factors like variations in its collection and handling. Initial findings have disclosed that also bacteria are crucial for the successful establishment of Candida in biofilms, but the clinical significance of this observation is yet to be confirmed. In conclusion, there is a need to standardize experimental procedures, to bridge the gap between laboratory and in vivo methodologies and findings and - in general - to thoroughly investigate the factors that modulate the initial attachment and subsequent colonization of denture-base materials and the oral mucosa of patients subjected to Candida infections. Information on how

  15. Enzymatic antioxidants status in patients with recurrent aphthous stomatitis.

    Science.gov (United States)

    Zhang, Zichuan; Li, Shan; Fang, Huiqing

    2017-10-01

    Oxidative stress (OS) has been thought to play a main role in the etiopathogenesis of recurrent aphthous stomatitis (RAS), which is one of the most common oral mucosal diseases characterized by recurrent and painful oral ulcers. The aim of this investigation was to evaluate the enzymatic antioxidants status in patients with RAS in the active stage and remission stage. Ninety-seven patients with idiopathic minor RAS and 102 race-, age- and gender-matched healthy individuals were recruited. All these subjects were allocated to three groups: RAS patients with active lesion (group A); the same patients in group A in the remission stage of RAS (group B); and healthy individuals without RAS (group C). Following an overnight fast, blood samples were obtained. The serum levels of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSHPx) were measured by the spectrophotometric method. Independent-samples t-test and paired t-test were performed for statistical evaluation. The serum levels of SOD, GSHPx, and CAT (83.9 ± 17.1 U/ml, 6687.2 ± 2629.2 U/ml, 1789.7 ± 593.8 U/l) were found to be significantly lower in group A as compared to those of group B (99.8 ± 11.1 U/ml, 9364.1 ± 1607.9 U/ml, 2789.1 ± 1113.4 U/l; P 0.05). Our results indicate that enzymatic antioxidant defense system is impaired in RAS patients with active lesion and seems to play a crucial role in its pathogenesis. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  16. Silver nitrate cautery in aphthous stomatitis: a randomized controlled trial.

    Science.gov (United States)

    Alidaee, M R; Taheri, A; Mansoori, P; Ghodsi, S Z

    2005-09-01

    Aphthous stomatitis is a painful, recurrent disease of the oral mucous membrane. Silver nitrate sticks have been used for a long time to provide pain relief for the duration of an aphthous ulceration, with only one application. Silver nitrate causes chemical cauterization and increases the depth of injury. To study the effect of chemical cautery with silver nitrate in reducing pain of aphthous ulceration and to determine if this treatment shortens or prolongs healing. In a randomized, patient-blinded, placebo-controlled study, 97 patients with painful minor oral aphthous ulceration were randomized to receive silver nitrate cautery or placebo. The severity of pain was rated on a three-category scale (severe, mild, none) and was recorded each day until the seventh day after the procedure. The lesion size was recorded at the time of the procedure and on the seventh day afterwards. In the treatment group, the ulcer was gently painted with a silver nitrate stick until it turned white. In the placebo group, the ulcer was gently painted with a placebo stick. In the treatment group, 33 of 47 patients (70%) evaluated and in the placebo group, four of 38 patients (11%) evaluated had reduction in severity of pain 1 day after the procedure. The difference was statistically significant (P aphthous ulceration without significantly shortening or prolonging healing time. We did not observe any side-effects in our study. The effect is rapid and lasts for the duration of the lesion. The treatment is simple and cost-effective in patients with infrequent recurrences.

  17. Mucosal and salivary microbiota associated with recurrent aphthous stomatitis.

    Science.gov (United States)

    Kim, Yun-Ji; Choi, Yun Sik; Baek, Keum Jin; Yoon, Seok-Hwan; Park, Hee Kyung; Choi, Youngnim

    2016-04-01

    Recurrent aphthous stomatitis (RAS) is a common oral mucosal disorder of unclear etiopathogenesis. Although recent studies of the oral microbiota by high-throughput sequencing of 16S rRNA genes have suggested that imbalances in the oral microbiota may contribute to the etiopathogenesis of RAS, no specific bacterial species associated with RAS have been identified. The present study aimed to characterize the microbiota in the oral mucosa and saliva of RAS patients in comparison with control subjects at the species level. The bacterial communities of the oral mucosa and saliva from RAS patients with active lesions (RAS, n = 18 for mucosa and n = 8 for saliva) and control subjects (n = 18 for mucosa and n = 7 for saliva) were analyzed by pyrosequencing of the 16S rRNA genes. There were no significant differences in the alpha diversity between the controls and the RAS, but the mucosal microbiota of the RAS patients showed increased inter-subject variability. A comparison of the relative abundance of each taxon revealed decreases in the members of healthy core microbiota but increases of rare species in the mucosal and salivary microbiota of RAS patients. Particularly, decreased Streptococcus salivarius and increased Acinetobacter johnsonii in the mucosa were associated with RAS risk. A dysbiosis index, which was developed using the relative abundance of A. johnsonii and S. salivarius and the regression coefficients, correctly predicted 83 % of the total cases for the absence or presence of RAS. Interestingly, A. johnsonii substantially inhibited the proliferation of gingival epithelial cells and showed greater cytotoxicity against the gingival epithelial cells than S. salivarius. RAS is associated with dysbiosis of the mucosal and salivary microbiota, and two species associated with RAS have been identified. This knowledge may provide a diagnostic tool and new targets for therapeutics for RAS.

  18. A Retrospective Evaluation of Patients with Recurrent Aphthous Stomatitis

    Directory of Open Access Journals (Sweden)

    Filiz Topaloğlu Demir

    2017-03-01

    Full Text Available Aim: To determine the factors in the etiology of recurrent aphthous stomatitis (RAS and to evaluate patients in the terms of RAS associated systemic disorders especially Behçet’s disease. Methods: Patients with RAS, who were followed up in Bartın State Hospital Dermatology Clinic between July 2013 and April 2015, were retrospectively evaluated. Results: A total of 123 patients (86 female, 37 male were included in this study. Thirteen (106% patients were children. The mean age of patients was 34.5±14.7 years (range: 8-69 years. Minor aphthous somatitis was the most frequent clinical type (68.3%. Family history was positive in 52.8% of patients. The triggering factors in the etiology of RAS were stress (54.5%, trauma (40.2%, gingivitis (29.3%, food (9.8%, medicines (5.7%, menstruation in female patients (3.3%, and throat infections (2.4%. Nutritional deficiencies were found in 39% of patients. There was a statistically significant difference in attack frequency (p=0.017 and throat infection history (p=0.029 between adults and pediatric patients. Fourteen (11.4% patients were diagnosed with Behçet’s disease. When we compared the RAS patients diagnosed with Behçet’s disease and the other RAS patients, a significant difference was found in pathergy test (p<0.001 and ferritin levels (p=0.020. Conclusion: Patients with RAS should be followed up for a long time for systemic disorders, especially for Behçet’s disease, accompanying RAS.

  19. Mucosal Microbiome in Patients with Recurrent Aphthous Stomatitis

    Science.gov (United States)

    Hijazi, K.; Lowe, T.; Meharg, C.; Berry, S.H.; Foley, J.; Hold, G.L.

    2015-01-01

    Recurrent aphthous stomatitis (RAS) is the most common disease affecting oral mucosae. Etiology is unknown, but several factors have been implicated, all of which influence the composition of microbiota residing on oral mucosae, which in turn modulates immunity and thereby affects disease progression. Although no individual pathogens have been conclusively shown to be causative agents of RAS, imbalanced composition of the oral microbiota may play a key role. In this study, we sought to determine composition profiles of bacterial microbiota in the oral mucosa associated with RAS. Using high-throughput 16S rRNA gene sequencing, we characterized the most abundant bacterial populations residing on healthy and ulcerated mucosae in patients with RAS (recruited using highly stringent criteria) and no associated medical conditions; we also compared these to the bacterial microbiota of healthy controls (HCs). Phylum-level diversity comparisons revealed decreased Firmicutes and increased Proteobacteria in ulcerated sites, as compared with healthy sites in RAS patients, and no differences between RAS patients with healthy sites and HCs. Genus-level analysis demonstrated higher abundance of total Bacteroidales in RAS patients with healthy sites over HCs. Porphyromonadaceae comprising species associated with periodontal disease and Veillonellaceae predominated in ulcerated sites over HCs, while no quantitative differences of these families were observed between healthy sites in RAS patients and HCs. Streptococcaceae comprising species associated with oral health predominated in HCs over ulcerated sites but not in HCs over healthy sites in RAS patients. This study demonstrates that mucosal microbiome changes in patients with idiopathic RAS—namely, increased Bacteroidales species in mucosae of RAS patients not affected by active ulceration. While these changes suggest a microbial role in initiation of RAS, this study does not provide data on causality. Within this limitation

  20. Clinical assessment of disease severity in recurrent aphthous stomatitis.

    Science.gov (United States)

    Tappuni, Anwar R; Kovacevic, Tatjana; Shirlaw, Penelope J; Challacombe, Stephen J

    2013-09-01

    Recurrent aphthous stomatitis (RAS) is one of the most common oral mucosal diseases in many parts of the world. However, there is very limited published clinical evidence for the therapies used in this condition. This could be partly due to the difficulty in evaluating the efficacy of oral ulcer treatment objectively. In this paper, we present a method for assessing and monitoring the severity of oral ulcers before and after treatment. Six ulcer characteristics, number, size, duration, ulcer-free period, site and pain, were used to generate an ulcer severity score (USS). The scores for 223 RAS patients were determined, and 79 were scored again after 3-month therapy with topical betamethasone. The scores for the minor RAS group were between 18 and 43 (mean 29.2 ± 5.3). The mean score in the major ulcers group (range: 28-60, mean 39.9 ± 6.1) was significantly greater than in the minor group (P < 0.001). The herpetiform recurrent ulcers score range was wide (range: 18-57, mean 36.6 ± 8.4). The mean severity score decreased significantly after treatment (P < 0.001). The USS was indicative of the disease activity in recurrent oral ulceration. It helped in assessing the efficacy of therapy, as the change in the numerical score reflected the change in ulcer severity in response to treatment. This tool may well prove to be of value in clinical management, research and in clinical trials. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  1. Periodic fever associated with aphthous stomatitis, pharyngitis and cervical adenitis.

    Science.gov (United States)

    Kyvsgaard, Nini; Mikkelsen, Torben; Korsholm, Jakob; Veirum, Jens Erik; Herlin, Troels

    2012-07-01

    The periodic fever, aphthous stomatitis, pharyngitis and cervical adenitis (PFAPA) syndrome is a non-hereditary idiopathic febrile syndrome belonging to the group of autoinflammatory diseases. PFAPA does not cause long-lasting sequelae. An early diagnosis provides treatment possibilities for the patient and comfort to the family. This study is a retrospective review of the medical records of patients diagnosed with PFAPA and admitted to our clinic from January 1999 to January 2010 (n = 31). The study population (n = 31) consisted of 21 males and ten females: 30 Caucasians and 1 Asian. Normal growth was seen in 30 patients. The median age at onset was 33 months. The mean duration of fever episodes was 4.45 days (95% confidence interval (CI): 3.92-4.98 days), and the mean duration of intervals between fever episodes was 29.66 days (95% CI: 25.31-34.01 days). Concomitantly with the fever, all patients had characteristic symptoms. All patients were asymptomatic in between their fever episodes. Prodromal symptoms were seen in 12 patients. Oral prednisolone was used in 24 patients and caused immediate fever reduction in 87.5%. A reduction in the duration of the asymptomatic interval after treatment was seen in 75.0%. Tonsillectomy was performed in 20 of the 31 patients causing cessation of fever episodes in 70%. Fever episodes continued in 15%, and the postoperative status remained unknown in the last 15%. Spontaneous resolution was seen in four patients. The diagnostic delay had a median duration of 28 months (range 2-160 months). The long diagnostic delay of PFAPA gives cause for concern and it indicates a need for greater awareness of the disease so that the diagnosis may be made earlier. not relevant. not relevant.

  2. Stomatal characterization of five species of the genus Vanilla.

    Directory of Open Access Journals (Sweden)

    Delfino Reyes-López

    2015-06-01

    Full Text Available The objective was to characterize the stomata of five species of vanilla. Throughout 2012, leaf samples of V. planifolia G. Jackson, V. pompona Schiede, V. indora Schiede, V. insignis Ames and V. odorota Presl were taken from the vanilla germplasm bank at the Benemérita Universidad Autónoma de Puebla. The stomata size was obtained considering their length and width, as well as the index and stomata number of the abaxial and adaxial leaf surfaces in a randomized complete block design with three replications. V. pompona Schiede and V. inodora Schiede showed the highest stomatal index with 8713 and 8246 stomata per mm2, respectively, followed by V. odorata Presl with 4412 stomata per mm2. V. insignis Ames and V. planifolia G. Jackson showed the lowest stomata index with 2968 and 1378 stomata per mm2, respectively, in the abaxial leaf surface, these differences were statistically significant (P≤0.05. According to the position of the leaf stomata, V. planifolia G. Jackson and V. inodora Schiede can be considered to be hypostomatics since they showed stomata only in the abaxial leaf surface. V. insignis Ames, V. inodora Schiede and V. odorata Presl. can be considered to be anfiestomatic because they showed stomata in both the abaxial and adaxial leaf surfaces. V. inodora Schiede had smaller stomata compared with the other species.That is an important feature to be included in the genetic improvement of the genus Vanilla, because due to climate change, temperature will increase and precipitation will decrease, so Vainilla will require more efficient genotypes for water use.

  3. Sociodemographic and clinical characteristics of patients with recurrent aphthous stomatitis

    Directory of Open Access Journals (Sweden)

    Anıl Gülsel Bahalı

    2014-12-01

    Full Text Available Background and Design: The purpose of this study was to obtain data that may provide an insight into the etiopathogenesis of recurrent aphtous stomatitis (RAS by the way of analysing the sociodemographic and clinical characteristics of patients who had been diagnosed with RAS. Materials and Metods: The patients, who were diagnosed with RAS in the dermatology outpatient clinic, between May 2007 and May 2010, were evaluated retrospectively. The data including sociodemografic and clinical characteristics, and treatment options were recorded. Results: A hundred patients (68 women, 32 men were included in this study. The average age was 40±13.6 years. RAS was more common in patients with middle-income and low education. The most common type of RAS was minor aphtous ulcers (88%. The lesions were most frequently seen on the lateral side of the tongue (34% and cheek (34%. Sixty percent of patients had a positive family history. Some factors such as biting (12%, tooth brushing (18%, dental disease presence (82%, food (39%, menstruation (10.3%, stress (76%, iron deficiency (16.7%, vitamin B12 deficiency (22.4%, low serum ferritin levels (18%, and seasonal variability (32% showed positive correlation with RAS. A negative correlation was found between RAS and smoking. Forty-nine percent of patients had used alternative therapies in addition to drug therapy. The most frequently used alternative method was consumption of sumac (26.5%. Conlucions: In contrast to the literature, our study found that RAS is started in the third decade of life and, approximately 50% of patients prefered alternative treatment methods, particularly sumac. Nowadays, discussions about the etiopathogenesis of RAS continue. In this study, we found that different sociodemographic and clinical factors may be associated with the etiopathogenesis of the disease. Our study will be followed by further studies using prospective design to identify the the etiopathogenesis of RAS.

  4. Modelling stomatal conductance in Acacia caven: A two way approach to understand vapor fluxes

    Science.gov (United States)

    Raab, N.; Meza, F. J.

    2012-12-01

    Evapotranspiration fluxes from semi arid ecosystems show a strong interannual variability and dependence on water availability. Usually this variable is regarded as very small but at local scale could substantially affect water balance at basin level. Climate Change scenarios for these regions are a source of concern as they project an increase in temperature, leading to a greater atmospheric water demand. In addition, precipitation is expected to decrease, increasing pressure for this kind of ecosystems. At a plant level, a rise on the actual atmospheric CO2 concentration is expected to improve photosynthetic performance and water use efficiency. However, as stomatal conductance is the main pathway for water vapor flux, from the leaf to the atmosphere, and CO2 entrance to the substomatal cavity, a larger control of the stomatal opening, due to a severe water control lost from the plant, could lead to shortages in net assimilation, jeopardizing the behavior of Semi Arid ecosystems as natural carbon sinks. Stoma is also one of the main lock of the soil-plant-water continuum, thus finally controlling the rate of soil water depletion. Its modeling presents a key role in determining future groundwater availability and net ecosystem exchange. There are several approaches for stomatal conductance modeling, from mechanistic models, based on the physiological functioning of the stomata, to empirical models where the stomatal behavior is correlated with environmental conditions. We modeled stomatal conductance for a Chilean typical Mediterranean Savannanh, dominated by Acacia caven, comparing two different empirical approaches. We used a Shuttleworth and Wallace model for sparse canopies combined with an inversion of the Penman-Monteith equation. This model allowed us to link stomatal conductance to evapotranspiration. The second approach was based on a multiplicative model for stomatal conductance based on environmental limitation, following Jarvis's model

  5. Salivary mucin MUC7 oligosaccharides in patients with recurrent aphthous stomatitis.

    Science.gov (United States)

    Zad, Mikael; Flowers, Sarah A; Bankvall, Maria; Jontell, Mats; Karlsson, Niclas G

    2015-11-01

    The aetiology of recurrent aphthous stomatitis remains unknown. In this study, we investigate the composition of oligosaccharides from mucin MUC7 in recurrent aphthous stomatitis as these heavily O-glycosylated mucins confer many of saliva's protective properties such as defence against mucosal pathogens. Unstimulated whole saliva samples were collected from six individuals, three with recurrent aphthous stomatitis and three corresponding sibling, without this condition. Oligosaccharides from salivary MUC7 were isolated and analysed by liquid chromatography-tandem mass spectrometry. The types of oligosaccharides identified in the patients and control subjects were similar; however, statistical evaluation indicated semi-quantitative differences between specific oligosaccharide classes. These changes focused on a reduction in terminal glycan residues including fucosylation, sialylation and sulfation on galactose. This study was able to show differential MUC7 glycosylation in the patients suggesting functional changes to salivary mucins in this condition. The terminal glycans altered in disease have been shown to be important for a range of immunological and bacterial binding roles. Further investigation of these epitopes in a larger study may provide critical insights into the pathology of recurrent aphthous stomatitis. MUC7 glycosylation is altered in recurrent aphthous stomatitis. This may change the protective properties of this mucin against mucosal pathogens, which may effect this condition.

  6. Correlation between histocompatibility antigens and recurrent aphthous stomatitis in the Brazilian population.

    Science.gov (United States)

    Wilhelmsen, Niels Salles Willo; Weber, Raimar; Monteiro, Francisco; Kalil, Jorge; Miziara, Ivan Dieb

    2009-01-01

    Recurrent aphthous stomatitis is a common oral mucosa disorder that affects 20% of the world's population, characterized by recurring painful ulcers in the mouth. The diagnosis is primarily based on the patient's clinical history. Inheritance may pose as a risk factor for the disease; however, the studies available are inconclusive as to the results attained, and they vary according to the population studied. to typify class I and class II HLA molecules and to assess how frequent these molecules are present in the Brazilian population with Recurrent Aphthous Stomatitis, compared to healthy controls. In this prospective, cross-sectional and investigative study, thirty one patients with diagnostic hypothesis of recurrent aphthous stomatitis were seen from February of 2004 to May of 2006. We obtained the DNA from those patients who matched the inclusion criteria and typified their HLA by PCR. In those patients with Recurrent Minor Aphthous Stomatitis we found statistically significant occurrences of HLA-A33 and HLA-B35. HLA-A33 and HLA-B35 may be associated with recurrent minor aphthous stomatitis in the Brazilian's population.

  7. INTERMEDIATE UVEITIS ASSOCIATED WITH PERIODIC FEVER, APHTHOUS STOMATITIS, PHARYNGITIS, AND CERVICAL ADENITIS SYNDROME.

    Science.gov (United States)

    Choi, Rene Y; Shakoor, Akbar; Bohnsack, John; Vitale, Albert T

    2017-05-29

    To report two novel cases of intermediate uveitis associated with Periodic Fever, Aphthous Stomatitis, Pharyngitis, and Cervical Adenitis syndrome. Observational case reports and review of the literature. Both patients in this report had an established diagnosis of Periodic Fever, Aphthous Stomatitis, Pharyngitis, and Cervical Adenitis syndrome before the onset of ocular inflammation. Infectious and noninfectious systemic conditions known to be associated with intermediate uveitis were excluded. Intermediate uveitis was confirmed clinically in both patients by the presence of vitritis, snowballs, and peripheral snowbanks in the region of the pars plana. Both cases had a course characterized by recurrent inflammation; in which systemic steroid treatment, and in one case, immunomodulatory therapy was necessary. Periodic Fever, Aphthous Stomatitis, Pharyngitis, and Cervical Adenitis syndrome is an auto-inflammatory fever disorder in childhood. Although other auto-inflammatory disorders such as, Blau syndrome, Muckle-Wells syndrome, and Behcets disease have been associated with various forms of uveitis, Periodic Fever, Aphthous Stomatitis, Pharyngitis, and Cervical Adenitis has never been reported to be associated with any type of ocular inflammation. We describe for the first time, two cases of Periodic Fever, Aphthous Stomatitis, Pharyngitis, and Cervical Adenitis syndrome presenting with intermediate uveitis.

  8. Zinc deficiency in patients with recurrent aphthous stomatitis: a pilot study.

    Science.gov (United States)

    Ozler, G S

    2014-06-01

    Recurrent aphthous stomatitis is a common disease of the oral mucosa, affecting 20 per cent of the general population.1 However, the aetiology of this disease is unknown. This is the first controlled study to compare zinc levels in recurrent aphthous stomatitis patients with those of a well-matched, healthy, control population. Twenty-five patients with a history of recurrent aphthous stomatitis and 25 healthy people (control group) took part in the study. Patients aged between 20 and 40 years with recurrent oral aphthous ulcers less than 1 cm in diameter were included. Exclusion criteria were pregnancy and lactation, systemic disease (ulcerative colitis, Crohn's disease, Behçet's disease), any current medication (topical or systemic), dental surgery during the previous month and deficiencies in iron, folic acid or vitamin B1, B2, B6 or B12. Serum zinc levels were compared between patient and control groups. Zinc deficiency was detected in 28 per cent of recurrent aphthous stomatitis patients and in 4 per cent of controls. The mean serum zinc level in the patient group was significantly lower than in the control group. These results suggest an association between zinc deficiency and recurrent aphthous stomatitis.

  9. Carbonic anhydrases are upstream regulators of CO2-controlled stomatal movements in guard cells

    KAUST Repository

    Hu, Honghong

    2009-12-13

    The continuing rise in atmospheric CO2 causes stomatal pores in leaves to close and thus globally affects CO2 influx into plants, water use efficiency and leaf heat stress. However, the CO2-binding proteins that control this response remain unknown. Moreover, which cell type responds to CO2, mesophyll or guard cells, and whether photosynthesis mediates this response are matters of debate. We demonstrate that Arabidopsis thaliana double-mutant plants in the beta-carbonic anhydrases betaCA1 and betaCA4 show impaired CO2-regulation of stomatal movements and increased stomatal density, but retain functional abscisic-acid and blue-light responses. betaCA-mediated CO2-triggered stomatal movements are not, in first-order, linked to whole leaf photosynthesis and can function in guard cells. Furthermore, guard cell betaca-overexpressing plants exhibit instantaneous enhanced water use efficiency. Guard cell expression of mammalian alphaCAII complements the reduced sensitivity of ca1 ca4 plants, showing that carbonic anhydrase-mediated catalysis is an important mechanism for betaCA-mediated CO2-induced stomatal closure and patch clamp analyses indicate that CO2/HCO3- transfers the signal to anion channel regulation. These findings, together with ht1-2 (ref. 9) epistasis analysis demonstrate that carbonic anhydrases function early in the CO2 signalling pathway, which controls gas-exchange between plants and the atmosphere.

  10. Differential Effects of Ozone Exposure on Carbon Assimilation and Stomatal Conductance

    Science.gov (United States)

    Lombardozzi, D.; Bonan, G. B.; Levis, S.; Sparks, J. P.

    2009-12-01

    Humans are indirectly increasing concentrations of surface ozone through industrial processes. Ozone is known to have negative impacts on plants, including reductions in crop yields, plant growth, and visible leaf injury. Ozone directly influences photosynthesis via two mechanisms: 1) the oxidation of cellular components (i.e., influencing leaf internal biochemistry and transport) and 2) altering stomatal functioning, ultimately changing conductance. Carbon exchange at the leaf level is governed by both conductance and carboxylation processes, but water exchange depends primarily on the size of the stomatal aperture. Thus, the possibility exists that ozone exposure will differentially affect plant-mediated carbon and water fluxes. Further, these differential effects of ozone are not explicitly expressed in most modeling efforts. We investigated how ozone changes both stomatal conductance and carbon assimilation using controlled open-top chamber experiments and then incorporated our experimental findings into modified Farquhar and Ball-Berry based photosynthesis and stomatal conductance models. In experiments, we observed carbon assimilation and conductance decreases in response to ozone. However, the decrease in carbon assimilation was larger than the decrease in conductance to water vapor, thereby changing the relationship between carbon gain and water loss at the leaf level. In addition, the relationship between photosynthesis and transpiration weakened significantly after 12 weeks of ozone exposure, suggesting a decoupling of photosynthesis and stomatal conductance. We used this information to modify biochemical parameters in the Farquhar model and the Ball-Berry coefficient to determine whether these models are able to simulate plant performance under ozone exposure.

  11. Exposure to moderate concentrations of tropospheric ozone impairs tree stomatal response to carbon dioxide

    Energy Technology Data Exchange (ETDEWEB)

    Onandia, Gabriela [Department of Plant and Environmental Sciences, University of Gothenburg, P. O. Box 461, SE-405 30 Goeteborg (Sweden); Cavanilles Institute of Biodiversity and Evolutionary Biology, Department of Microbiology and Ecology, University of Valencia, E-46100 Burjassot, Valencia (Spain); Olsson, Anna-Karin; Barth, Sabine [Department of Plant and Environmental Sciences, University of Gothenburg, P. O. Box 461, SE-405 30 Goeteborg (Sweden); King, John S. [Department of Forestry and Environmental Resources, Campus Box 8002, North Carolina State University, Raleigh, NC 27695 (United States); Uddling, Johan, E-mail: johan.uddling@dpes.gu.se [Department of Plant and Environmental Sciences, University of Gothenburg, P. O. Box 461, SE-405 30 Goeteborg (Sweden)

    2011-10-15

    With rising concentrations of both atmospheric carbon dioxide (CO{sub 2}) and tropospheric ozone (O{sub 3}), it is important to better understand the interacting effects of these two trace gases on plant physiology affecting land-atmosphere gas exchange. We investigated the effect of growth under elevated CO{sub 2} and O{sub 3}, singly and in combination, on the primary short-term stomatal response to CO{sub 2} concentration in paper birch at the Aspen FACE experiment. Leaves from trees grown in elevated CO{sub 2} and/or O{sub 3} exhibited weaker short-term responses of stomatal conductance to both an increase and a decrease in CO{sub 2} concentration from current ambient level. The impairement of the stomatal CO{sub 2} response by O{sub 3} most likely developed progressively over the growing season as assessed by sap flux measurements. Our results suggest that expectations of plant water-savings and reduced stomatal air pollution uptake under rising atmospheric CO{sub 2} may not hold for northern hardwood forests under concurrently rising tropospheric O{sub 3}. - Exposure to moderate concentrations of tropospheric ozone impairs stomatal CO{sub 2} responsiveness of birch in the Aspen FACE experiment.

  12. Balancing Water Uptake and Loss through the Coordinated Regulation of Stomatal and Root Development.

    Directory of Open Access Journals (Sweden)

    Christopher Hepworth

    Full Text Available Root development is influenced by nutrient and water availabilities. Plants are able to adjust many attributes of their root in response to environmental signals including the size and shape of the primary root, lateral roots and root hairs. Here we investigated the response of roots to changes in the levels of leaf transpiration associated with altered stomatal frequency. We found that plants with high stomatal density and conductance produce a larger rooting area and as a result have enhanced phosphate uptake capacity whereas plants with low stomatal conductance produce a smaller root. Manipulating the growth environment of plants indicated that enhanced root growth is most likely a result of an increased demand for water rather than phosphate. Plants manipulated to have an increase or reduction in root hair growth show a reduction or increase respectively, in stomatal conductance and density. Our results demonstrate that plants can balance their water uptake and loss through coordinated regulation of both stomatal and root development.

  13. Regulation of acetylcholine receptor clustering by ADF/cofilin-directed vesicular trafficking.

    Science.gov (United States)

    Lee, Chi Wai; Han, Jianzhong; Bamburg, James R; Han, Liang; Lynn, Rachel; Zheng, James Q

    2009-07-01

    Postsynaptic receptor localization is crucial for synapse development and function, but the underlying cytoskeletal mechanisms remain elusive. Using Xenopus neuromuscular junctions as a model, we found that actin depolymerizing factor (ADF)/cofilin regulated actin-dependent vesicular trafficking of acetylcholine receptors (AChRs) to the postsynaptic membrane. Active ADF/cofilin was concentrated in small puncta adjacent to AChR clusters and was spatiotemporally correlated with the formation and maintenance of surface AChR clusters. Notably, increased actin dynamics, vesicular markers and intracellular AChRs were all enriched at the sites of ADF/cofilin localization. Furthermore, a substantial amount of new AChRs was detected at these ADF/cofilin-enriched sites. Manipulation of either ADF/cofilin activity through its serine-3 phosphorylation or ADF/cofilin localization via 14-3-3 proteins markedly attenuated AChR insertion and clustering. These results suggest that spatiotemporally restricted ADF/cofilin-mediated actin dynamics regulate AChR trafficking during the development of neuromuscular synapses.

  14. Ethosomes: versatile vesicular carriers for efficient transdermal delivery of therapeutic agents.

    Science.gov (United States)

    Pandey, Vikas; Golhani, Dilip; Shukla, Rajesh

    2015-12-01

    Delivery across skin is attractive due to its easy accessibility. However, drug delivery across skin is still a challenge in biomedical sciences. Over the past few decades, various successful novel devices and techniques have emerged to optimize drug delivery across skin whose obstructing behavior constricts entry of most of the therapeutic agents. Inability of various conventional vesicular formulations, e.g. liposomes to pass through the tapered (>30 nm) intercellular channels of stratum corneum, rendered invention of some lipid based vesicular carrier systems such as ethosomes which consist of phospholipid, ethanol and water. Ethosomes are non-invasive delivery carriers that enable drugs to reach the deep skin layers and/or the systemic circulation. In spite of their sophistication in conceptuality, they are exemplified by easiness in their preparation, safety and efficacy - a combination that can highly inflate their application. This review attempts to describe all aspects of ethosomes including roles and upshots of different excipients, various methods of preparation and characterizations, research reports on various drug deliveries, patent reports and future prospects.

  15. 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

    2017-03-04

    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 3 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 uses 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.

  16. Vesicular calcium regulates coat retention, fusogenicity, and size of pre-Golgi intermediates.

    Science.gov (United States)

    Bentley, Marvin; Nycz, Deborah C; Joglekar, Ashwini; Fertschai, Ismene; Malli, Roland; Graier, Wolfgang F; Hay, Jesse C

    2010-03-15

    The significance and extent of Ca(2+) regulation of the biosynthetic secretory pathway have been difficult to establish, and our knowledge of regulatory relationships integrating Ca(2+) with vesicle coats and function is rudimentary. Here, we investigated potential roles and mechanisms of luminal Ca(2+) in the early secretory pathway. Specific depletion of luminal Ca(2+) in living normal rat kidney cells using cyclopiazonic acid (CPA) resulted in the extreme expansion of vesicular tubular cluster (VTC) elements. Consistent with this, a suppressive role for vesicle-associated Ca(2+) in COPII vesicle homotypic fusion was demonstrated in vitro using Ca(2+) chelators. The EF-hand-containing protein apoptosis-linked gene 2 (ALG-2), previously implicated in the stabilization of sec31 at endoplasmic reticulum exit sites, inhibited COPII vesicle fusion in a Ca(2+)-requiring manner, suggesting that ALG-2 may be a sensor for the effects of vesicular Ca(2+) on homotypic fusion. Immunoisolation established that Ca(2+) chelation inhibits and ALG-2 specifically favors residual retention of the COPII outer shell protein sec31 on pre-Golgi fusion intermediates. We conclude that vesicle-associated Ca(2+), acting through ALG-2, favors the retention of residual coat molecules that seem to suppress membrane fusion. We propose that in cells, these Ca(2+)-dependent mechanisms temporally regulate COPII vesicle interactions, VTC biogenesis, cargo sorting, and VTC maturation.

  17. GLTP mediated non-vesicular GM1 transport between native membranes.

    Directory of Open Access Journals (Sweden)

    Ines Lauria

    Full Text Available Lipid transfer proteins (LTPs are emerging as key players in lipid homeostasis by mediating non-vesicular transport steps between two membrane surfaces. Little is known about the driving force that governs the direction of transport in cells. Using the soluble LTP glycolipid transfer protein (GLTP, we examined GM1 (monosialotetrahexosyl-ganglioside transfer to native membrane surfaces. With artificial GM1 donor liposomes, GLTP can be used to increase glycolipid levels over natural levels in either side of the membrane leaflet, i.e., external or cytosolic. In a system with native donor- and acceptor-membranes, we find that GLTP balances highly variable GM1 concentrations in a population of membranes from one cell type, and in addition, transfers lipids between membranes from different cell types. Glycolipid transport is highly efficient, independent of cofactors, solely driven by the chemical potential of GM1 and not discriminating between the extra- and intracellular membrane leaflet. We conclude that GLTP mediated non-vesicular lipid trafficking between native membranes is driven by simple thermodynamic principles and that for intracellular transport less than 1 µM GLTP would be required in the cytosol. Furthermore, the data demonstrates the suitability of GLTP as a tool for artificially increasing glycolipid levels in cellular membranes.

  18. Vibrio tapetis isolated from vesicular skin lesions in Dover sole Solea solea.

    Science.gov (United States)

    Declercq, A M; Chiers, K; Soetaert, M; Lasa, A; Romalde, J L; Polet, H; Haesebrouck, F; Decostere, A

    2015-06-29

    Vibrio tapetis is primarily known as the causative agent for brown ring disease in bivalves, although it has been isolated from cultivated fish during mortalities on farms. Here we describe the first isolation of V. tapetis from wild-caught and subsequently captive-held Dover sole Solea solea. Pathological features consisted of multifocal circular greyish-white skin discolourations evolving into vesicular lesions and subsequent ulcerations on the pigmented side. On the non-pigmented side, multiple circular lesions-white at the center and red at the edges-were evident. Histological examination of the vesicular lesions revealed dermal fluid-filled spaces, collagen tissue necrosis and a mixed inflammatory infiltrate, with large numbers of small rod-shaped bacteria. In the deep skin lesions, loss of scales and dermal connective tissue, with degeneration and fragmentation of the myofibres bordering the ulceration, were noted. Serotyping, DNA-DNA hybridization and REP- and ERIC-PCR techniques showed that the retrieved isolates displayed a profile similar to the representative strain of genotype/serotype O2 which originally was isolated from carpet-shell clam Venerupis decussata and to which isolates obtained from wedge sole Dicologoglossa cuneata were also closely related.

  19. Orf virus interferes with MHC class I surface expression by targeting vesicular transport and Golgi

    Directory of Open Access Journals (Sweden)

    Rohde Jörg

    2012-07-01

    Full Text Available Abstract Background The Orf virus (ORFV, a zoonotic Parapoxvirus, causes pustular skin lesions in small ruminants (goat and sheep. Intriguingly, ORFV can repeatedly infect its host, despite the induction of a specific immunity. These immune modulating and immune evading properties are still unexplained. Results Here, we describe that ORFV infection of permissive cells impairs the intracellular transport of MHC class I molecules (MHC I as a result of structural disruption and fragmentation of the Golgi apparatus. Depending on the duration of infection, we observed a pronounced co-localization of MHC I and COP-I vesicular structures as well as a reduction of MHC I surface expression of up to 50%. These subversion processes are associated with early ORFV gene expression and are accompanied by disturbed carbohydrate trimming of post-ER MHC I. The MHC I population remaining on the cell surface shows an extended half-life, an effect that might be partially controlled also by late ORFV genes. Conclusions The presented data demonstrate that ORFV down-regulates MHC I surface expression in infected cells by targeting the late vesicular export machinery and the structure and function of the Golgi apparatus, which might aid to escape cellular immune recognition.

  20. Stomatal and non-stomatal limitations on leaf carbon assimilation in beech (Fagus sylvatica L.) seedlings under natural conditions

    Energy Technology Data Exchange (ETDEWEB)

    Aranda, I.; Rodriguez-Calcerrada, J.; Robson, T. M.; Cano, F. J.; Alte, L.; Sanchez-Gomez, D.

    2012-07-01

    Limitations to diffusion and biochemical factors affecting leaf carbon uptake were analyzed in young beech seedlings (Fagus sylvtica L.) growing in natural gaps of a beech-wood at the southern limit of the species. Half of the seedlings received periodic watering in addition to natural rainfall to reduce the severity of the summer drought. Plant water status was evaluated by measuring predawn water potential. Basic biochemical parameters were inferred from chlorophyll fluorescence and photosynthesis-CO{sub 2} curves (A-C{sub c}) under saturating light. The curves were established on three dates during the summer months. The main variables studied included: stomatal and mesophyll conductance to CO{sub 2} (g{sub s} and g{sub m} respectively), maximum velocity of carboxylation (V{sub c}max) and maximum electron transport capacity (J{sub m}ax). The gm was estimated by two methodologies: the curve-fitting and J constant methods. Seedlings withstood moderate water stress, as the leaf predawn water potential ({Psi}{sub p}d) measured during the study was within the range -0.2 to -0.5 MPa. Mild drought caused gs and gm to decrease only slightly in response to {Psi}{sub p}d. However both diffusional parameters explained most of the limitations to CO{sub 2} uptake. In addition, it should be highlighted that biochemical limitations, prompted by V{sub c}max and J{sub m}ax, were related mainly to ontogenic factors, without any clear relationship with drought under the moderate water stress experienced by beech seedlings through the study. The results may help to further understanding of the functional mechanisms influencing the carbon fixation capacity of beech seedlings under natural conditions. (Author) 68 refs.

  1. Fístula colecistoduodenal, complicación infrecuente de litiasis vesicular: nuestra experiencia en su manejo quirúrgico

    OpenAIRE

    F. Aguilar-Espinosa; R. Maza-Sánchez; F. Vargas-Solís; G.A. Guerrero-Martínez; J.L. Medina-Reyes; P.I. Flores-Quiroz

    2017-01-01

    Introducción: Las fístulas bilioentéricas son la comunicación anormal entre el sistema biliar y el tracto gastrointestinal, que ocurre de manera espontánea y en la mayoría de los casos es una complicación rara de la litiasis vesicular no tratada. Pueden provocar consecuencias clínicas diversas que, en algunas situaciones, ponen en peligro la vida del paciente. Objetivo: Identificar la incidencia de fístula bilioentérica en pacientes con litiasis vesicular, su presentación clínica, diagnóst...

  2. Didelphis marsupialis como un reservorio potencial u hospedero amplificador del virus de la estomatitis vesicular, serotipo new jersey en Antioquia

    OpenAIRE

    John Arboleda; Carlos Trujillo

    2004-01-01

    La Estomatitis Vesicular (EV) es una enfermedad viral, aguda
    y autolimitante que afecta principalmente bovinos, equinos y
    porcinos. Es producida por el virus de estomatitis vesicular (VEV), serotipos New Jersey (VEV-NJ) e Indiana (VEV-IN), que son los as importantes epidemiológicamente (1). Los estudios serológicos demuestran que VEV-NJ y VEV-IN infectan en forma natural una gran variedad de animales silvestres, que están posiblemente implicados en la  coepizoot...

  3. Investigations on a Flaps-Like Disease of Cattle (’Benign Flaps,’ Stomatitis Papulosa Bovis Specifica) (Untersuchungen uber eine Maulseucheahnliche Erkrankung des Rindes (’Gutartige Maulseuche,’ Stomatitis Papulosa Bovis Specifica))

    Science.gov (United States)

    Stomatitis papulosa infections of cattle can be differentiated from so-called sporadic aphtha and from aphthous fever. Stomatitis papulosa can be...distinguished from both diseases by the absence of blisters and pustules. Moreover, it can also be differentiated from aphthous fever in that it involves only the oral cavity and not the skin and hooves.

  4. Oral symptoms and salivary findings in oral lichen planus, oral lichenoid lesions and stomatitis

    DEFF Research Database (Denmark)

    Larsen, Kristine Roen; Johansen, Jeanne Duus; Reibel, Jesper

    2017-01-01

    BACKGROUND: To examine if patients with oral lichen planus, oral lichenoid lesions and generalised stomatitis and concomitant contact allergy have more frequent and severe xerostomia, lower unstimulated and chewing-stimulated saliva and citric-acid-stimulated parotid saliva flow rates, and higher...... of xerostomia, clinical examination, sialometry, mucosal biopsy and contact allergy testing. RESULTS: Nineteen patients had oral lichen planus, 19 patients had oral lichenoid lesions and 11 patients had generalised stomatitis. 38.8% had contact allergy. Xerostomia was significantly more common and severe...... in the chewing stimulated saliva samples from patients when compared to healthy controls. The differences were not significant and they were irrespective of the presence of contact allergy. CONCLUSION: Xerostomia is prevalent in patients with oral lichen planus, lichenoid lesions and generalised stomatitis...

  5. Oral symptoms and salivary findings in oral lichen planus, oral lichenoid lesions and stomatitis

    DEFF Research Database (Denmark)

    Larsen, Kristine Røn; Johansen, Jeanne Duus; Reibel, Jesper

    2017-01-01

    Background: To examine if patients with oral lichen planus, oral lichenoid lesions and generalised stomatitis and concomitant contact allergy have more frequent and severe xerostomia, lower unstimulated and chewing-stimulated saliva and citric-acid-stimulated parotid saliva flow rates, and higher......, clinical examination, sialometry, mucosal biopsy and contact allergy testing. Results: 19 patients had oral lichen planus, 19 patients had oral lichenoid lesions and 11 patients had generalised stomatitis. 38.8% had contact allergy. Xerostomia was significantly more common and severe in patients (46...... in the chewing stimulated saliva samples from patients when compared to healthy controls. The differences were not significant and they were irrespective of the presence of contact allergy. Conclusion: Xerostomia is prevalent in patients with oral lichen planus, lichenoid lesions and generalised stomatitis...

  6. Ulcerative Uremic Stomatitis - Review of the Literature and A Rare Case Report

    Directory of Open Access Journals (Sweden)

    Shantala Arunkumar

    2015-01-01

    Full Text Available Uremic Stomatitis (US represents a comparatively uncommon intraoral complication seen, mostly, in cases of end-stage renal disease or undiagnosed or untreated chronic renal failure. Its frequency has diminished due to the advent of renal dialysis. Clinically uremic stomatitis is characterized by the presence of painful plaques and crusts that are usually distributed on the buccal and labial mucosa, dorsal or ventral surface of the tongue, gingiva, and floor of the mouth. Ultimate treatment consists of improvement of blood urea concentration and underlying renal failure is supported by enhancement of oral hygiene with antiseptic mouthwashes and antimicrobial/antifungal agents, if necessary. Here we report a rare case of ulcerative type of uremic stomatitis occurring in a patient of chronic renal failure due to sudden relapse of uremia and reviewed the possible pathophysiology of oral symptoms of chronic renal failure.

  7. Guard cell sensory systems: recent insights on stomatal responses to light, abscisic acid, and CO2.

    Science.gov (United States)

    Assmann, Sarah M; Jegla, Timothy

    2016-10-01

    By controlling the opening and closure of the stomatal pores through which gas exchange occurs, guard cells regulate two of the most important plant physiological processes: photosynthesis and transpiration. Accordingly, guard cells have evolved exquisite sensory systems. Here we summarize recent literature on guard cell sensing of light, drought (via the phytohormone abscisic acid (ABA)), and CO2. New advances in our understanding of how guard cells satisfy the energetic and osmotic requirements of stomatal opening and utilize phosphorylation to regulate the anion channels and aquaporins involved in ABA-stimulated stomatal closure are highlighted. Omics and modeling approaches are providing new information that will ultimately allow an integrated understanding of guard cell physiology. Copyright © 2016. Published by Elsevier Ltd.

  8. Gluten sensitivity enteropathy in patients with recurrent aphthous stomatitis

    Directory of Open Access Journals (Sweden)

    Davatchi Fereydoun

    2009-06-01

    Full Text Available Abstract Background Gluten sensitive enteropathy (GSE is an autoimmune enteropathy triggered by the ingestion of gluten-containing grains in susceptible individuals. Recurrent aphthous stomatitis (RAS may be the sole manifestation of GSE. The aim of this study was to determine the prevalence of gluten sensitivity enteropathy (GSE in a large group of patients with RAS and assess the efficacy of gluten free diet (GFD on the improvement of aphthous lesions in those who were diagnosed with GSE. Methods Two hundred and forty seven patients with RAS were included. The patients had at least three aphthous attacks per year. Patients were screened by IgA anti-endomysial antibody (EMA, IgA anti tissue transglutaminase (TTG and serum IgA level. Those with a positive serology underwent endoscopic biopsies of the duodenal mucosa and patients with negative serology were excluded. The diagnosis of GSE was based on a positive serological test and abnormal duodenal histology. For patients with GSE, gluten free diet was recommended. Results Six out of 247 RAS patients had positive TTG test alone, and one had positive EMA and TTG. All 7 patients with positive serologic tests underwent duodenal biopsies. Histological findings were compatible with GSE in all of them (Marsh I in four patients, Marsh II in two patients and Marsh IIIB in one another.. The mean age of GSE patients was 27.42 ± 10.56 (range, 13 to 40 years old. They were suffering from RAS for an average duration of 4.5 years. All of the 7 GSE patients had not responded to the routine anti-aphthae medications, including topical corticosteroids, tetracycline and colchicine. Four patients who adhered to a strict gluten-free diet showed noticeable improvement in their aphthous lesions over a period of 6 months. Conclusion A significant minority (e.g. 2.83% of RAS patients have GSE. This could be compared with the 0.9% prevalence of GSE in the general population of Iran. This study suggests that evaluation

  9. Canopy Stomatal Conductance Unlocks Partitioning of Ecosystem-Atmosphere Carbon and Water Exchanges

    Science.gov (United States)

    Wehr, R. A.; Munger, J. W.; McManus, J. B.; Nelson, D. D.; Zahniser, M. S.; Davidson, E. A.; Wofsy, S. C.; Saleska, S. R.

    2016-12-01

    Stomata are a key nexus in biosphere-atmosphere interactions: the gateway for both carbon gain and water loss by plant canopies. Accurate quantification of canopy stomatal conductance enables partitioning of both evapotranspiration (ET) and net ecosystem-atmosphere CO2 exchange (NEE)—the latter via CO2 isotope flux measurements. To those ends, we determined the behavior of canopy stomatal conductance in a temperate deciduous forest based on heat and water vapor flux measurements, and validated that determination based on uptake of carbonyl sulfide, which also passes through the stomata. We found that the canopy stomatal conductance followed a simple empirical function of leaf area index, light intensity, diffuse light fraction, and leaf-air water vapor gradient. The dependence on light intensity was highly linear, in contrast to the leaf scale, and in contrast to the behavior of canopy photosynthesis. Using canopy stomatal conductance, we partitioned ET and found that evaporation in this ecosystem peaks at the time of the year when soils are driest and atmospheric vapor pressure deficit is low—because soil temperature is an important driver. As stomatal conductance impacts not only the rate of photosynthesis but also the fractionation of carbon isotopes by photosynthesis, we were also able to combine canopy stomatal conductance with CO2 isotope flux measurements in order to partition NEE. We found that: (1) canopy respiration is much less during the day than at night, likely due to the inhibition of leaf respiration by light (that is, the Kok effect), and (2) canopy photosynthetic light-use efficiency does not decline through the summer, in contrast to standard estimates. These results clarify how leaf-level physiological dynamics impact ecosystem-atmosphere gas exchange, and demonstrate the utility of combining multiple tracers to constrain the processes underlying that exchange.

  10. Carbon and hydrogen isotopic effects of stomatal density in Arabidopsis thaliana

    Science.gov (United States)

    Lee, Hyejung; Feakins, Sarah J.; Sternberg, Leonel da S. L.

    2016-04-01

    Stomata are key gateways mediating carbon uptake and water loss from plants. Varied stomatal densities in fossil leaves raise the possibility that isotope effects associated with the openness of exchange may have mediated plant wax biomarker isotopic proxies for paleovegetation and paleoclimate in the geological record. Here we use Arabidopsis thaliana, a widely used model organism, to provide the first controlled tests of stomatal density on carbon and hydrogen isotopic compositions of cuticular waxes. Laboratory grown wildtype and mutants with suppressed and overexpressed stomatal densities allow us to directly test the isotope effects of stomatal densities independent of most other environmental or biological variables. Hydrogen isotope (D/H) measurements of both plant waters and plant wax n-alkanes allow us to directly constrain the isotopic effects of leaf water isotopic enrichment via transpiration and biosynthetic fractionations, which together determine the net fractionation between irrigation water and n-alkane hydrogen isotopic composition. We also measure carbon isotopic fractionations of n-alkanes and bulk leaf tissue associated with different stomatal densities. We find offsets of +15‰ for δD and -3‰ for δ13C for the overexpressed mutant compared to the suppressed mutant. Since the range of stomatal densities expressed is comparable to that found in extant plants and the Cenozoic fossil record, the results allow us to consider the magnitude of isotope effects that may be incurred by these plant adaptive responses. This study highlights the potential of genetic mutants to isolate individual isotope effects and add to our fundamental understanding of how genetics and physiology influence plant biochemicals including plant wax biomarkers.

  11. Stomatal kinetics and photosynthetic gas exchange along a continuum of isohydric to anisohydric regulation of plant water status

    Science.gov (United States)

    Frederick C. Meinzer; Duncan D. Smith; David R. Woodruff; Danielle E. Marias; Katherine A. McCulloh; Ava R. Howard; Alicia L. Magedman

    2017-01-01

    Species’ differences in the stringency of stomatal control of plant water potential represent a continuum of isohydric to anisohydric behaviours. However, little is known about how quasi-steady-state stomatal regulation of water potential may relate to dynamic behaviour of stomata and photosynthetic gas exchange in species operating at different positions along this...

  12. Hypofunction of the Sympathetic Nervous System as a Possible Etiologic Cause of Recurrent Aphthous Stomatitis.

    Science.gov (United States)

    Present, Steven I; Check, Jerome H

    2016-06-01

    Recurrent aphthous stomatitis is a common disorder of the oral mucosa. The symptoms can range from a minor nuisance to severe forms that can be extremely debilitating. Two cases of chronic aphthous stomatitis are described. The patients sought help to ameliorate vasomotor symptoms. A diagnosis of sympathetic nervous system hypofunction was established. Treatment was aimed at restoring normal sympathetic function by the administration of dextroamphetamine sulfate. Since the patients have been on the amphetamine salts, neither their vasomotor symptoms nor their aphthous lesions have returned. Hypofunction of the sympathetic nervous system should be considered as a possible etiologic factor in patients with recurrent oral ulcers when not associated with known systemic diseases.

  13. Heterotrimeric G protein mediates ethylene-induced stomatal closure via hydrogen peroxide synthesis in Arabidopsis.

    Science.gov (United States)

    Ge, Xiao-Min; Cai, Hong-Li; Lei, Xue; Zhou, Xue; Yue, Ming; He, Jun-Min

    2015-04-01

    Heterotrimeric G proteins function as key players in hydrogen peroxide (H2O2) production in plant cells, but whether G proteins mediate ethylene-induced H2O2 production and stomatal closure are not clear. Here, evidences are provided to show the Gα subunit GPA1 as a missing link between ethylene and H2O2 in guard cell ethylene signalling. In wild-type leaves, ethylene-triggered H2O2 synthesis and stomatal closure were dependent on activation of Gα. GPA1 mutants showed the defect of ethylene-induced H2O2 production and stomatal closure, whereas wGα and cGα overexpression lines showed faster stomatal closure and H2O2 production in response to ethylene. Ethylene-triggered H2O2 generation and stomatal closure were impaired in RAN1, ETR1, ERS1 and EIN4 mutants but not impaired in ETR2 and ERS2 mutants. Gα activator and H2O2 rescued the defect of RAN1 and EIN4 mutants or etr1-3 in ethylene-induced H2O2 production and stomatal closure, but only rescued the defect of ERS1 mutants or etr1-1 and etr1-9 in ethylene-induced H2O2 production. Stomata of CTR1 mutants showed constitutive H2O2 production and stomatal closure, but which could be abolished by Gα inhibitor. Stomata of EIN2, EIN3 and ARR2 mutants did not close in responses to ethylene, Gα activator or H2O2, but do generate H2O2 following challenge of ethylene or Gα activator. The data indicate that Gα mediates ethylene-induced stomatal closure via H2O2 production, and acts downstream of RAN1, ETR1, ERS1, EIN4 and CTR1 and upstream of EIN2, EIN3 and ARR2. The data also show that ETR1 and ERS1 mediate both ethylene and H2O2 signalling in guard cells. © 2015 The Authors The Plant Journal © 2015 John Wiley & Sons Ltd.

  14. Plasticity in stomatal size and density of potato leaves under different irrigation and phosphorus regimes

    DEFF Research Database (Denmark)

    Sun, Yanqi; Yan, Fei; Cui, Xiaoyong

    2014-01-01

    The morphological features of stomata including their size and density could be modulated by environmental cues; however, the underlying mechanisms remain largely elusive. Here, the effect of different irrigation and phosphorus (P) regimes on stomatal size (SS) and stomatal density (SD) of potato...... leaves was investigated. The plants were grown in split-root pots under two P fertilization rates (viz., 0 and 100mgkg-1 soil, denoted as P0 and P1, respectively) and subjected to full (FI), deficit (DI), and partial root-zone drying (PRD) irrigation regimes. Results showed that SS and SD were...

  15. Stomatal Conductance, Plant Species Distribution, and an Exploration of Rhizosphere Microbes and Mycorrhizae at a Deliberately Leakimg Experimental Carbon Sequestration Field (ZERT)

    Science.gov (United States)

    Sharma, B.; Apple, M. E.; Morales, S.; Zhou, X.; Holben, B.; Olson, J.; Prince, J.; Dobeck, L.; Cunningham, A. B.; Spangler, L.

    2010-12-01

    One measure to reduce atmospheric CO2 is to sequester it in deep geological formations. Rapid surface detection of any CO2 leakage is crucial. CO2 leakage rapidly affects vegetation above sequestration fields. Plant responses to high CO2 are valuable tools in surface detection of leaking CO2. The Zero Emission Research Technology (ZERT) site in Bozeman, MT is an experimental field for surface detection of CO2 where 0.15 ton/day of CO2 was released (7/19- 8/15/2010) from a 100m horizontal injection well, HIW, 1.5 m underground with deliberate leaks of CO2 at intervals, and from a vertical injector, VI, (6/3-6/24/2010). The vegetation includes Taraxacum officinale (Dandelion), Dactylis glomerata (Orchard Grass), and other herbaceous plants. We collected soil and roots 1, 3 and 5 m from the VI to determine the responses of mycorrhizal fungi and rhizosphere microbes to high CO2. Mycorrhizal fungi obtain C from root exudates, increase N and P availability, and reduce desiccation, while prokaryotic rhizosphere microbes fix atmospheric N and will be examined for abundance and expression of carbon and nitrogen cycling genes. We are quantifying mycorrhizal colonization and the proportion of spores, hyphae, and arbuscules in vesicular-arbuscular mycorrhizae (VAM) in cleared and stained roots. Stomatal conductance is an important measure of CO2 uptake and water loss via transpiration. We used a porometer (5-40°C, 0-90% RH, Decagon) to measure stomatal conductivity in dandelion and orchard grass at 1, 3, and 5 m from the VI and along a transect perpendicular to the HIW. Dandelion conductance was highest close to the VI and almost consistently higher close to hot spots (circular regions with maximum CO2 and leaf dieback) at the HIW, with 23.2 mmol/m2/s proximal to the hot spot, and 10.8 mmol/m2/s distally. Average conductance in grass (50.3 mmol/m2/s) was higher than in dandelion, but grass did not have high conductance near hot spots. Stomata generally close at elevated CO2

  16. Impaired stomatal control is associated with reduced photosynthetic physiology in crop species grown at elevated [CO2

    Directory of Open Access Journals (Sweden)

    Matthew Haworth

    2016-10-01

    Full Text Available Physiological control of stomatal conductance (Gs permits plants to balance CO2-uptake for photosynthesis (PN against water-loss, so optimising water use efficiency (WUE. An increase in the atmospheric concentration of carbon dioxide ([CO2] will result in a stimulation of PN and reduction of Gs in many plants, enhancing carbon gain while reducing water-loss. It has also been hypothesised that the increase in WUE associated with lower Gs at elevated [CO2] would reduce the negative impacts of drought on many crops. Despite the large number of CO2-enrichment studies to date, there is relatively little information regarding the effect of elevated [CO2] on stomatal control. Five crop species with active physiological stomatal behaviour were grown at ambient (400 ppm and elevated (2000 ppm [CO2]. We investigated the relationship between stomatal function, stomatal size and photosynthetic capacity in the five species, and then assessed the mechanistic effect of elevated [CO2] on photosynthetic physiology, stomatal sensitivity to [CO2] and the effectiveness of stomatal closure to darkness. We observed positive relationships between the speed of stomatal response and the maximum rates of PN and Gs sustained by the plants; indicative of close co-ordination of stomatal behaviour and PN. In contrast to previous studies we did not observe a negative relationship between speed of stomatal response and stomatal size. The sensitivity of stomata to [CO2] declined with the ribulose-1,5-bisphosphate limited rate of PN at elevated [CO2]. The effectiveness of stomatal closure was also impaired at high [CO2]. Growth at elevated [CO2] did not affect the performance of photosystem II indicating that high [CO2] had not induced damage to the photosynthetic physiology, and suggesting that photosynthetic control of Gs is either directly impaired at high [CO2], sensing/signalling of environmental change is disrupted or elevated [CO2] causes some physical effect that

  17. Carbon fiber ultramicrodic electrode electrodeposited with over-oxidized polypyrrole for amperometric detection of vesicular exocytosis from pheochromocytoma cell.

    Science.gov (United States)

    Wang, Li; Xu, Huiren; Song, Yilin; Luo, Jinping; Xu, Shengwei; Zhang, Song; Liu, Juntao; Cai, Xinxia

    2015-01-06

    Vesicular exocytosis is ubiquitous, but it is difficult to detect within the cells' communication mechanism. For this purpose, a 2 µm ultramicrodic carbon fiber electrode was fabricated in this work based on electrodeposition with over-oxidized polypyrrole nanoparticle (PPyox-CFE), which was applied successfully for real-time monitoring of quantal exocytosis from individual pheochromocytoma (PC12) cells. PPyox-CFE was evaluated by dopamine (DA) solutions through cyclic voltammetry and amperometry electrochemical methods, and results revealed that PPyox-CFE improved the detection limit of DA. In particular, the sensitivity of DA was improved to 24.55 µA·µM(-1)·µm(-2) using the PPyox-CFE. The ultramicrodic electrode combined with the patch-clamp system was used to detect vesicular exocytosis of DA from individual PC12 cells with 60 mM K+ stimulation. A total of 287 spikes released from 7 PC12 cells were statistically analyzed. The current amplitude (Imax) and the released charge (Q) of the amperometric spikes from the DA release by a stimulated PC12 cell is 45.1 ± 12.5 pA and 0.18 ± 0.04 pC, respectively. Furthermore, on average ~562,000 molecules were released in each vesicular exocytosis. PPyox-CFE, with its capability of detecting vesicular exocytosis, has potential application in neuron communication research.

  18. Carbon Fiber Ultramicrodic Electrode Electrodeposited with Over-Oxidized Polypyrrole for Amperometric Detection of Vesicular Exocytosis from Pheochromocytoma Cell

    Directory of Open Access Journals (Sweden)

    Li Wang

    2015-01-01

    Full Text Available Vesicular exocytosis is ubiquitous, but it is difficult to detect within the cells’ communication mechanism. For this purpose, a 2 µm ultramicrodic carbon fiber electrode was fabricated in this work based on electrodeposition with over-oxidized polypyrrole nanoparticle (PPyox-CFE, which was applied successfully for real-time monitoring of quantal exocytosis from individual pheochromocytoma (PC12 cells. PPyox-CFE was evaluated by dopamine (DA solutions through cyclic voltammetry and amperometry electrochemical methods, and results revealed that PPyox-CFE improved the detection limit of DA. In particular, the sensitivity of DA was improved to 24.55 µA·µM−1·µm−2 using the PPyox-CFE. The ultramicrodic electrode combined with the patch-clamp system was used to detect vesicular exocytosis of DA from individual PC12 cells with 60 mM K+ stimulation. A total of 287 spikes released from 7 PC12 cells were statistically analyzed. The current amplitude (Imax and the released charge (Q of the amperometric spikes from the DA release by a stimulated PC12 cell is 45.1 ± 12.5 pA and 0.18 ± 0.04 pC, respectively. Furthermore, on average ~562,000 molecules were released in each vesicular exocytosis. PPyox-CFE, with its capability of detecting vesicular exocytosis, has potential application in neuron communication research.

  19. Contribution of vesicular and cytosolic dopamine to the increased striatal dopamine efflux elicited by intrastriatal injection of dexamphetamine.

    NARCIS (Netherlands)

    Watanabe, S.; Aono, Y.; Fusa, K.; Takada, K.; Saigusa, T.; Koshikawa, N.; Cools, A.R.

    2005-01-01

    Systemic administration of high doses of dexamphetamine induces a dopamine efflux that has its intracellular origin in both the vesicular, reserpine-sensitive dopamine pool and the cytosolic, alpha-methyl-para-tyrosine-sensitive, newly synthesized dopamine pool. It remains unknown whether locally

  20. Contribution of vesicular and cytosolic dopamine to the increased striatal dopamine efflux elicited by intrastriatal injection of SKF38393.

    NARCIS (Netherlands)

    Saigusa, T.; Aono, Y.; Sekino, R.; Uchida, T.; Takada, K.; Oi, Y.; Koshikawa, N.; Cools, A.R.

    2009-01-01

    Like dexamphetamine, SKF38393 induces an increase in striatal dopamine efflux which is insensitive for tetrodotoxin, Ca(2+) independent and prevented by a dopamine transporter inhibitor. The dexamphetamine-induced striatal dopamine efflux originates from both the reserpine-sensitive vesicular

  1. Reduction of bacterial growth by a vesicular-arbuscular mycorrhizal fungus in the rhizosphere of cucumber (Cucumis sativus L.)

    DEFF Research Database (Denmark)

    Christensen, H.; Jakobsen, I.

    1993-01-01

    Cucumber was grown in a partially sterilized sand-soil mixture with the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus fasciculatum or left uninoculated. Fresh soil extract was places in polyvinyl chloride tubes without propagules of mycorrhizal fungi. Root tips and root segments...

  2. Carbon Fiber Ultramicrodic Electrode Electrodeposited with Over-Oxidized Polypyrrole for Amperometric Detection of Vesicular Exocytosis from Pheochromocytoma Cell

    Science.gov (United States)

    Wang, Li; Xu, Huiren; Song, Yilin; Luo, Jinping; Xu, Shengwei; Zhang, Song; Liu, Juntao; Cai, Xinxia

    2015-01-01

    Vesicular exocytosis is ubiquitous, but it is difficult to detect within the cells' communication mechanism. For this purpose, a 2 μm ultramicrodic carbon fiber electrode was fabricated in this work based on electrodeposition with over-oxidized polypyrrole nanoparticle (PPyox-CFE), which was applied successfully for real-time monitoring of quantal exocytosis from individual pheochromocytoma (PC12) cells. PPyox-CFE was evaluated by dopamine (DA) solutions through cyclic voltammetry and amperometry electrochemical methods, and results revealed that PPyox-CFE improved the detection limit of DA. In particular, the sensitivity of DA was improved to 24.55 μA·μM−1·μm−2 using the PPyox-CFE. The ultramicrodic electrode combined with the patch-clamp system was used to detect vesicular exocytosis of DA from individual PC12 cells with 60 mM K+ stimulation. A total of 287 spikes released from 7 PC12 cells were statistically analyzed. The current amplitude (Imax) and the released charge (Q) of the amperometric spikes from the DA release by a stimulated PC12 cell is 45.1 ± 12.5 pA and 0.18 ± 0.04 pC, respectively. Furthermore, on average ∼562,000 molecules were released in each vesicular exocytosis. PPyox-CFE, with its capability of detecting vesicular exocytosis, has potential application in neuron communication research. PMID:25569759

  3. APPARENT LACK OF VESICULAR-ARBUSCULAR MYCORRHIZA (VAM) IN SEAGRASSES ZOSTERA MARINA L. AND THALASSIA TESTUDIUM BANKS EX KONIG

    Science.gov (United States)

    We examined two populations of Zostera marina L. and one of Thalassia testudinum Banks ex Konig for presence of vesicular-arbuscular mycorrhiza (VAM). None of these plants showed any VAM colonization. In addition, we were unable to find any literature references on the presence o...

  4. Vesicular monoamine transporter 1 gene polymorphism and white matter integrity in major depressive disorder.

    Science.gov (United States)

    Won, Eunsoo; Han, Kyu-Man; Kang, June; Kim, Aram; Yoon, Ho-Kyoung; Chang, Hun Soo; Park, Ji-Young; Lee, Min-Soo; Greenberg, Tsafrir; Tae, Woo-Suk; Ham, Byung-Joo

    2017-07-03

    The genetic variant of the vesicular monoamine transporter 1 gene (VMAT1) has been suggested to be associated with monoaminergic signaling and neural circuit activity related to emotion processing. We aimed to investigate microstructural changes in white matter tracts of patients with major depressive disorder (MDD), and examined the interaction effect between VMAT1 Thr136Ile (rs1390938) polymorphism and MDD on white matter integrity. Diffusion tensor imaging (DTI) and VMAT1 Thr136Ile (rs1390938) genotyping were performed on 103 patients diagnosed with MDD and 83 healthy control participants. DTI was used to investigate microstructural changes in white matter tracts in patients compared to healthy controls. The possible interaction effect between rs1390938 and MDD on white matter integrity was also assessed. Patients with MDD exhibited lower fractional anisotropy (FA) values of the forceps major (pdepression. Copyright © 2017 Elsevier Inc. All rights reserved.

  5. Polysaccharide nano-vesicular multidrug carriers for synergistic killing of cancer cells

    Science.gov (United States)

    Pramod, P. S.; Shah, Ruchira; Chaphekar, Sonali; Balasubramanian, Nagaraj; Jayakannan, Manickam

    2014-09-01

    Multi-drug delivery based on polymer nano-scaffolds is an essential protocol to be developed for better administration of anticancer drugs to enhance their therapeutic efficacies against cancer cells. Here, we report dual delivery polysaccharide nano-vesicles that are capable of loading and delivering both water soluble and water insoluble drugs together in a single polymer scaffold. The selective rupture of the nano-vesicular assembly under intracellular enzyme conditions allowed the simultaneous delivery of a hydrophobic drug camptothecin (CPT) and hydrophilic drug doxorubicin (DOX) supporting their synergistic killing of breast and colon cancer cells. The polysaccharide nano-vesicles have allowed us to address a few important questions regarding the need for multiple drug administration in cancer cells including (a) the role of simultaneous drug release, (b) antagonistic versus synergistic effects of drug combinations and (c) how these are affected by the ratio of drugs. Further, evaluation of the role of caveolae in endocytosis of these polymer scaffolds was also made. The vesicular scaffolds were found to preserve and deliver DOX resulting in 50-60% better killing of cancer cells than the free drug. Additionally, dual loaded nano-vesicles when compared to drug cocktails with individual drugs in separate nano-vesicles (at comparable molar ratios) suggest the relative drug concentration following release and mode of delivery to be both important in cancer cell killing. Results from these experiments have revealed newly developed polysaccharide nano-vesicles loaded with DOX and CPT drugs as potential candidates for improved breast cancer cell killing. Thus, these custom-designed polysaccharide nano-vesicles provide a new perspective on multi-anticancer drug delivery systems and their efficacy.Multi-drug delivery based on polymer nano-scaffolds is an essential protocol to be developed for better administration of anticancer drugs to enhance their therapeutic

  6. `Full fusion' is not ineluctable during vesicular exocytosis of neurotransmitters by endocrine cells

    Science.gov (United States)

    Oleinick, Alexander; Svir, Irina; Amatore, Christian

    2017-01-01

    Vesicular exocytosis is an essential and ubiquitous process in neurons and endocrine cells by which neurotransmitters are released in synaptic clefts or extracellular fluids. It involves the fusion of a vesicle loaded with chemical messengers with the cell membrane through a nanometric fusion pore. In endocrine cells, unless it closes after some flickering (`Kiss-and-Run' events), this initial pore is supposed to expand exponentially, leading to a full integration of the vesicle membrane into the cell membrane-a stage called `full fusion'. We report here a compact analytical formulation that allows precise measurements of the fusion pore expansion extent and rate to be extracted from individual amperometric spike time courses. These data definitively establish that, during release of catecholamines, fusion pores enlarge at most to approximately one-fifth of the radius of their parent vesicle, hence ruling out the ineluctability of `full fusion'.

  7. The Dynamic Control of Kiss-And-Run and Vesicular Reuse Probed with Single Nanoparticles

    Science.gov (United States)

    Zhang, Qi; Li, Yulong; Tsien, Richard W.

    2009-01-01

    Summary Vesicular secretion of neurotransmitter is essential for neuronal communication. Kiss-and-run is a mode of membrane fusion and retrieval without the full collapse of the vesicle into the plasma membrane and de novo regeneration. The significance of kiss-and-run during efficient neurotransmission has remained in doubt. We developed an approach for loading individual synaptic vesicles with single quantum dots. Their size and pH-dependent photoluminescence change allowed us to distinguish kiss-and-run from full-collapse fusion and to track single vesicles through multiple rounds of kiss-and-run and reuse, without perturbing vesicle cycling. Kiss-and-run dominated at the beginning of stimulus trains, reflecting the preference of vesicles with high release probability. Its incidence was increased by rapid firing, a response appropriate to meet the dynamic demands of neurotransmission. PMID:19213879

  8. Unusual armadillo fold in the human general vesicular transport factor p115.

    Directory of Open Access Journals (Sweden)

    Harald Striegl

    Full Text Available The golgin family gives identity and structure to the Golgi apparatus and is part of a complex protein network at the Golgi membrane. The golgin p115 is targeted by the GTPase Rab1a, contains a large globular head region and a long region of coiled-coil which forms an extended rod-like structure. p115 serves as vesicle tethering factor and plays an important role at different steps of vesicular transport. Here we present the 2.2 A-resolution X-ray structure of the globular head region of p115. The structure exhibits an armadillo fold that is decorated by elongated loops and carries a C-terminal non-canonical repeat. This terminal repeat folds into the armadillo superhelical groove and allows homodimeric association with important implications for p115 mediated multiple protein interactions and tethering.

  9. Pesquisaje de litiasis vesicular en un sector de población supuestamente sana

    OpenAIRE

    Lázaro Yera Abreus; Mercedes Cárdenas Drake; Angel Gutiérrez Rojas

    1997-01-01

    Se hace un estudio de frecuencia de litiasis vesicular en un sector de población supuestamente sana, en el que se encontró una frecuencia de la afección de un 6,2 %, el predominio de las personas de la raza blanca sobre las de la raza negra fue de sólo 1,8:1; la afección es mucho más frecuente en el sexo femenino que en el masculino (proporción de 9:1) y en personas mayores de 40 años (60 %) con sobrepeso u obesas (85 %), y puede cursar de forma totalmente asintomática en el 50 % de los casos...

  10. BIOCHEMICAL CHARACTERIZATION OF NATIVE USHER PROTEIN COMPLEXES FROM A VESICULAR SUBFRACTION OF TRACHEAL EPITHELIAL CELLS†

    Science.gov (United States)

    Zallocchi, Marisa; Sisson, Joseph H.; Cosgrove, Dominic

    2010-01-01

    Usher syndrome is the major cause of deaf/blindness in the world. It is a genetic heterogeneous disorder, with nine genes already identified as causative for the disease. We noted expression of all known Usher proteins in bovine tracheal epithelial cells, and exploited this system for large-scale biochemical analysis of Usher protein complexes. The dissected epithelia were homogenized in non-detergent buffer, and sedimented on sucrose gradients. At least two complexes were evident after the first gradient: one formed by specific isoforms of CDH23, PCDH15 and VLGR-1, and a different one at the top of the gradient that included all the Usher proteins and rab5, a transport vesicle marker. TEM analysis of these top fractions found them enriched in 100–200 nm vesicles, confirming a vesicular association of the Usher complex(es). Immunoisolation of these vesicles confirmed some of the associations already predicted and identified novel interactions. When the vesicles are lysed in the presence of phenylbutyrate, most of the Usher proteins co-sediment into the gradient at a sedimentation coefficient of approximately 50S, correlating with a predicted molecular mass of 2 × 106 Daltons. Although it is still unclear whether there is only one complex or several independent complexes that are trafficked within distinct vesicular pools, this work shows for the first time that native Usher proteins complexes occur in vivo. This complex(es) is present primarily in transport vesicles at the apical pole of tracheal epithelial cells, predicting that Usher proteins may be directionally transported as complexes in hair cells and photoreceptors. PMID:20058854

  11. Topical vesicular formulations of Curcuma longa extract on recuperating the ultraviolet radiation-damaged skin.

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    Kaur, Chanchal Deep; Saraf, Swarnlata

    2011-12-01

      Ultraviolet radiations generate reactive oxygen species, leading to adverse effects on skin properties. Botanical extracts are multifunctional in nature having various properties like photoprotection, anti-aging, moisturizing, antioxidant, astringent, anti-irritant, and antimicrobial activity.   The aim of this study was to formulate creams having Curcuma longa extract loaded novel vesicular systems (liposomes, ethosomes, and transfersomes) and study their photoprotective effect by assessment of skin hydration (Cutometer) and sebum content (Sebumeter).   The alcoholic C. longa extract loaded liposomes, ethosomes, and transfersomes having 0.5-2.0% w/w extract were prepared, evaluated for size, entrapment efficiency, and incorporated into the cream. Their long-term interaction with skin (6 weeks) was compared in terms of their effects on skin hydration and sebum content.   Vesicular size obtained was in the range 167.3 ± 3.0 to 262.4 ± 2.4 nm with low polydispersity index (0.2-0.3) and high entrapment efficiency. The efficacy was in the order C. longa extract loaded transfersomal creams > C. longa extract loaded ethosomal creams > C. longa extract loaded liposomal creams > C. longa extract loaded creams > Empty transfersome loaded cream > Empty ethosome loaded cream > Empty liposome loaded cream > Base cream.   The photoprotective properties of the constituents of C. longa extract and hydrant, moisturizing lipid components of nano vesicles with better skin penetration resulted in improvement in skin properties like skin hydration and sebum content. The herbal extract loaded nano vesicles incorporated in cream could be used as photoprotective formulations. © 2011 Wiley Periodicals, Inc.

  12. Potential fossil endoliths in vesicular pillow basalt, Coral Patch Seamount, eastern North Atlantic Ocean.

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    Cavalazzi, Barbara; Westall, Frances; Cady, Sherry L; Barbieri, Roberto; Foucher, Frédéric

    2011-09-01

    The chilled rinds of pillow basalt from the Ampère-Coral Patch Seamounts in the eastern North Atlantic were studied as a potential habitat of microbial life. A variety of putative biogenic structures, which include filamentous and spherical microfossil-like structures, were detected in K-phillipsite-filled amygdules within the chilled rinds. The filamentous structures (∼2.5 μm in diameter) occur as K-phillipsite tubules surrounded by an Fe-oxyhydroxide (lepidocrocite) rich membranous structure, whereas the spherical structures (from 4 to 2 μm in diameter) are associated with Ti oxide (anatase) and carbonaceous matter. Several lines of evidence indicate that the microfossil-like structures in the pillow basalt are the fossilized remains of microorganisms. Possible biosignatures include the carbonaceous nature of the spherical structures, their size distributions and morphology, the presence and distribution of native fluorescence, mineralogical and chemical composition, and environmental context. When taken together, the suite of possible biosignatures supports the hypothesis that the fossil-like structures are of biological origin. The vesicular microhabitat of the rock matrix is likely to have hosted a cryptoendolithic microbial community. This study documents a variety of evidence for past microbial life in a hitherto poorly investigated and underestimated microenvironment, as represented by the amygdules in the chilled pillow basalt rinds. This kind of endolithic volcanic habitat would have been common on the early rocky planets in our Solar System, such as Earth and Mars. This study provides a framework for evaluating traces of past life in vesicular pillow basalts, regardless of whether they occur on early Earth or Mars.

  13. Motor dysfunction in cerebellar Purkinje cell-specific vesicular GABA transporter knockout mice

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    Mikiko eKayakabe

    2014-01-01

    Full Text Available γ-Aminobutyric acid (GABA is a major inhibitory neurotransmitter in the adult mammalian central nervous system and plays modulatory roles in neural development. The vesicular GABA transporter (VGAT is an essential molecule for GABAergic neurotransmission due to its role in vesicular GABA release. Cerebellar Purkinje cells (PCs are GABAergic projection neurons that are indispensable for cerebellar function. To elucidate the significance of VGAT in cerebellar PCs, we generated and characterized PC-specific VGAT knockout (L7-VGAT mice. VGAT mRNAs and proteins were specifically absent in the 40-week-old L7-VGAT PCs. The morphological charactereistics, such as lamination and foliation of the cerebellar cortex, of the L7-VGAT mice were similar to those of the control littermate mice. Moreover, the protein expression levels and patterns of pre- (calbindin and parvalbumin and postsynaptic (GABA-A receptor α1 subunit (GABAARα1 and gephyrin molecules between the L7-VGAT and control mice were similar in the deep cerebellar nuclei that receive PC projections. However, the L7-VGAT mice performed poorly in the accelerating rotarod test and displayed ataxic gait in the footprint test. The L7-VGAT mice also exhibited severer ataxia as VGAT deficits progressed. These results suggest that VGAT in cerebellar Purkinje cells is not essential for the rough maintenance of cerebellar structure, but does play an important role in motor coordination. The L7-VGAT mice are a novel model of ataxia without PC degeneration, and would also be useful for studying the role of Purkinje cells in cognition and emotion.

  14. Molecular mechanisms of Sar/Arf GTPases in vesicular trafficking in yeast and plants.

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    Yorimitsu, Tomohiro; Sato, Ken; Takeuchi, Masaki

    2014-01-01

    Small GTPase proteins play essential roles in the regulation of vesicular trafficking systems in eukaryotic cells. Two types of small GTPases, secretion-associated Ras-related protein (Sar) and ADP-ribosylation factor (Arf), act in the biogenesis of transport vesicles. Sar/Arf GTPases function as molecular switches by cycling between active, GTP-bound and inactive, GDP-bound forms, catalyzed by guanine nucleotide exchange factors and GTPase-activating proteins, respectively. Activated Sar/Arf GTPases undergo a conformational change, exposing the N-terminal amphipathic α-helix for insertion into membranes. The process triggers the recruitment and assembly of coat proteins to the membranes, followed by coated vesicle formation and scission. In higher plants, Sar/Arf GTPases also play pivotal roles in maintaining the dynamic identity of organelles in the secretory pathway. Sar1 protein strictly controls anterograde transport from the endoplasmic reticulum (ER) through the recruitment of plant COPII coat components onto membranes. COPII vesicle transport is responsible for the organization of highly conserved polygonal ER networks. In contrast, Arf proteins contribute to the regulation of multiple trafficking routes, including transport through the Golgi complex and endocytic transport. These transport systems have diversified in the plant kingdom independently and exhibit several plant-specific features with respect to Golgi organization, endocytic cycling, cell polarity and cytokinesis. The functional diversification of vesicular trafficking systems ensures the multicellular development of higher plants. This review focuses on the current knowledge of Sar/Arf GTPases, highlighting the molecular details of GTPase regulation in vesicle formation in yeast and advances in knowledge of the characteristics of vesicle trafficking in plants.

  15. Molecular mechanisms of Sar/Arf GTPases in vesicular trafficking in yeast and plants

    Directory of Open Access Journals (Sweden)

    Tomohiro eYorimitsu

    2014-08-01

    Full Text Available Small GTPase proteins play essential roles in the regulation of vesicular trafficking systems in eukaryotic cells. Two types of small GTPases, secretion-associated Ras-related protein (Sar and ADP-ribosylation factor (Arf, act in the biogenesis of transport vesicles. Sar/Arf GTPases function as molecular switches by cycling between active, GTP-bound and inactive, GDP-bound forms, catalyzed by guanine nucleotide exchange factors and GTPase-activating proteins, respectively. Activated Sar/Arf GTPases undergo a conformational change, exposing the N-terminal amphipathic α-helix for insertion into membranes. The process triggers the recruitment and assembly of coat proteins to the membranes, followed by coated vesicle formation and scission. In higher plants, Sar/Arf GTPases also play pivotal roles in maintaining the dynamic identity of organelles in the secretory pathway. Sar1 GTPase strictly controls anterograde transport from the endoplasmic reticulum (ER through the recruitment of plant COPII coat components onto membranes. COPII vesicle transport is responsible for the organization of highly conserved polygonal ER networks. In contrast, Arf GTPases contribute to the regulation of multiple trafficking routes, including transport through the Golgi complex and endocytic transport. These transport systems have diversified in the plant kingdom independently and exhibit several plant-specific features with respect to Golgi organization, endocytic cycling, cell polarity and cytokinesis. The functional diversification of vesicular trafficking systems ensures the multicellular development of higher plants. This review focuses on the current knowledge of Sar/Arf GTPases, highlighting the molecular details of GTPase regulation in vesicle formation in yeast and advances in knowledge of the characteristics of vesicle trafficking in plants.

  16. Developing ultra deformable vesicular transportation of a bioactive alkaloid in pursuit of vitiligo therapy

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    Vinod KR

    2012-08-01

    Full Text Available Objective: To develop transfersomal formulation integrated with piperine intended for vitiligo. Methods: Film hydration technique was employed in the preparation of transfersomes. Modified diffusion cell, consistency tester were fabricated for ex vivo diffusion studies and spreadability studies respectively while tape stripping method was integrated with tissue extraction in the determination of tissue drug concentration. Results: When film hydration technique was used for, ultradeformable vesicles (transfersomes of piperine in soabean phosphatidylcholine was formed with (67.11依0.22 to (70.55依3.62 and (60.12依1.04 to (80.43依0.14 mean size (毺 m and entrapment efficiency (% respectively. Transfersomes are capable of crossing the pores in permeability barriers extremely efficient even if the transfersome radius (tr is much greater than the pore size (rpore ie., tr/rpore曒0.25 the driven flux rate depends on the transdermal osmotic gradient. The vesicles describes elasto-mechanical character of vesicles while penetrating through the pores. The proviso is that the vesicular membrane elasticity is dynamically to the local stress by the external. Diffusion and Spreadability studies showed maximum diffusion when the lipid was kept minimum. Tape stripping and tissue extraction method for the tissue drug retention showed that (75.25依1.72% drug was retained in the dermis. Conclusions: Span 80 was preferred over tween 80 in terms of dermal retention. Size and encapsulation was slightly altered by phosphatidylcholine concentration. The kinetics, efficiency and the transfersome mediated transport can be tailored for trans-epidermal, deep tissues and systemic depending on the vesicular composition, dose and form. Thus we have offered a successful drug delivery of piperine targeting the deep epidermis.

  17. [A PERSPECTIVE CULTURAL MODEL FOR CONTROL OF BIOLOGICAL ACTIVITY OF HUMAN INTERFERONS].

    Science.gov (United States)

    Nagieva, F G; Barkova, E P; Fedotov, A Yu; Gaiderova, L A; Lisakov, A N; Nochevny, V T

    2015-01-01

    Study species specificity of human lymphocyte interferon alpha in vitro in cell cultures of swine origin for expansion of cell line spectrum for interferon titration and control of newly created interferons and interferon-like preparations in vivo in mini-pig model. Cell cultures of various species origin were used: Vero (monkey kidney), MDBK (bull kidney), HEK 293T (human embryo kidney), PK-15 (swine kidney), SPEV(swine embryo kidney), PTP (swine testicles), MDCK (canine kidney), RK-13 (rabbit kidney). Human lymphocyte interferon alpha (hINF-alpha) from Biomed company (1000 IU/ml), established in MDBK cells, was tested. Vesicular stomatitis virus (Indiana strain) was used. Human plasma was obtained from heparin-treated venous blood in the process of human peripheral blood lymphocyte isolation in medium for lymphocyte separation (Ficoll with a density of 1.077 g/cm3). Vesicular stomatitis virus, adapted to Vero cells, was established to have the least active reproduction in Vero and MDBK and reproduces more actively in cell of swine origin by 0.25 - 0.75 lg TCD50. At the same time, virus, adapted to cells of swine origin, reproduces more actively by 2 - 3 lg TCD50 in both cells of swine origin and Vero and MDBK. A possibility of titration of hINF-alpha in cells of swine origin was shown for both 100 doses of the indicator virus and low virus doses (5 and 10). This allows to determine low titers of hINF-alpha in blood plasma as one of the important indicators of interferon status--sera hINF-alpha.

  18. Results of the Study of Helminths-Carrying as a Comorbidity in Children with Herpetic Stomatitis

    Directory of Open Access Journals (Sweden)

    E.S. Suerkulov

    2016-11-01

    Full