Oncotargeting by Vesicular Stomatitis Virus (VSV: Advances in Cancer Therapy

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Suman Bishnoi

2018-02-01

Full Text Available Modern oncotherapy approaches are based on inducing controlled apoptosis in tumor cells. Although a number of apoptosis-induction approaches are available, site-specific delivery of therapeutic agents still remain the biggest hurdle in achieving the desired cancer treatment benefit. Additionally, systemic treatment-induced toxicity remains a major limiting factor in chemotherapy. To specifically address drug-accessibility and chemotherapy side effects, oncolytic virotherapy (OV has emerged as a novel cancer treatment alternative. In OV, recombinant viruses with higher replication capacity and stronger lytic properties are being considered for tumor cell-targeting and subsequent cell lysing. Successful application of OVs lies in achieving strict tumor-specific tropism called oncotropism, which is contingent upon the biophysical interactions of tumor cell surface receptors with viral receptors and subsequent replication of oncolytic viruses in cancer cells. In this direction, few viral vector platforms have been developed and some of these have entered pre-clinical/clinical trials. Among these, the Vesicular stomatitis virus (VSV-based platform shows high promise, as it is not pathogenic to humans. Further, modern molecular biology techniques such as reverse genetics tools have favorably advanced this field by creating efficient recombinant VSVs for OV; some have entered into clinical trials. In this review, we discuss the current status of VSV based oncotherapy, challenges, and future perspectives regarding its therapeutic applications in the cancer treatment.

Oncotargeting by Vesicular Stomatitis Virus (VSV): Advances in Cancer Therapy.

Science.gov (United States)

Bishnoi, Suman; Tiwari, Ritudhwaj; Gupta, Sharad; Byrareddy, Siddappa N; Nayak, Debasis

2018-02-23

Modern oncotherapy approaches are based on inducing controlled apoptosis in tumor cells. Although a number of apoptosis-induction approaches are available, site-specific delivery of therapeutic agents still remain the biggest hurdle in achieving the desired cancer treatment benefit. Additionally, systemic treatment-induced toxicity remains a major limiting factor in chemotherapy. To specifically address drug-accessibility and chemotherapy side effects, oncolytic virotherapy (OV) has emerged as a novel cancer treatment alternative. In OV, recombinant viruses with higher replication capacity and stronger lytic properties are being considered for tumor cell-targeting and subsequent cell lysing. Successful application of OVs lies in achieving strict tumor-specific tropism called oncotropism, which is contingent upon the biophysical interactions of tumor cell surface receptors with viral receptors and subsequent replication of oncolytic viruses in cancer cells. In this direction, few viral vector platforms have been developed and some of these have entered pre-clinical/clinical trials. Among these, the Vesicular stomatitis virus (VSV)-based platform shows high promise, as it is not pathogenic to humans. Further, modern molecular biology techniques such as reverse genetics tools have favorably advanced this field by creating efficient recombinant VSVs for OV; some have entered into clinical trials. In this review, we discuss the current status of VSV based oncotherapy, challenges, and future perspectives regarding its therapeutic applications in the cancer treatment.

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

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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.

Analysis of the RNA species isolated from defective particles of vesicular stomatitis virus.

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Adler, R; Banerjee, A K

1976-10-01

Serial high multiplicity passage of a cloned stock of vesicular stomatitis virus was found to generate defective interfering particles containing three size classes of RNA, with sedimentaiton coefficients of 31 S, 23 S and 19 S. The 31 S and 23 S RNA species were found to be complementary to both the 12 to 18 S and 31 S size classes of VSV mRNAs. The 19 S class of RNA was found to be partially base-paired. All three RNA species were found to contain ppAp at their 5' termini.

Anterograde or Retrograde Transsynaptic Circuit Tracing in Vertebrates with Vesicular Stomatitis Virus Vectors.

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Beier, Kevin T; Mundell, Nathan A; Pan, Y Albert; Cepko, Constance L

2016-01-04

Viruses have been used as transsynaptic tracers, allowing one to map the inputs and outputs of neuronal populations, due to their ability to replicate in neurons and transmit in vivo only across synaptically connected cells. To date, their use has been largely restricted to mammals. In order to explore the use of such viruses in an expanded host range, we tested the transsynaptic tracing ability of recombinant vesicular stomatitis virus (rVSV) vectors in a variety of organisms. Successful infection and gene expression were achieved in a wide range of organisms, including vertebrate and invertebrate model organisms. Moreover, rVSV enabled transsynaptic tracing of neural circuitry in predictable directions dictated by the viral envelope glycoprotein (G), derived from either VSV or rabies virus (RABV). Anterograde and retrograde labeling, from initial infection and/or viral replication and transmission, was observed in Old and New World monkeys, seahorses, jellyfish, zebrafish, chickens, and mice. These vectors are widely applicable for gene delivery, afferent tract tracing, and/or directional connectivity mapping. Here, we detail the use of these vectors and provide protocols for propagating virus, changing the surface glycoprotein, and infecting multiple organisms using several injection strategies. Copyright © 2016 John Wiley & Sons, Inc.

Long-Term Single-Dose Efficacy of a Vesicular Stomatitis Virus-Based Andes Virus Vaccine in Syrian Hamsters

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Joseph Prescott

2014-01-01

Full Text Available Andes virus (ANDV is highly pathogenic in humans and is the primary etiologic agent of hantavirus cardiopulmonary syndrome (HCPS in South America. Case-fatality rates are as high as 50% and there are no approved vaccines or specific therapies for infection. Our laboratory has recently developed a replication-competent recombinant vesicular stomatitis virus (VSV-based vaccine that expressed the glycoproteins of Andes virus in place of the native VSV glycoprotein (G. This vaccine is highly efficacious in the Syrian hamster model of HCPS when given 28 days before challenge with ANDV, or when given around the time of challenge (peri-exposure, and even protects when administered post-exposure. Herein, we sought to test the durability of the immune response to a single dose of this vaccine in Syrian hamsters. This vaccine was efficacious in hamsters challenged intranasally with ANDV 6 months after vaccination (p = 0.025, but animals were not significantly protected following 1 year of vaccination (p = 0.090. The decrease in protection correlated with a reduction of measurable neutralizing antibody responses, and suggests that a more robust vaccination schedule might be required to provide long-term immunity.

A Recombinant Vesicular Stomatitis Virus Ebola Vaccine.

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Regules, Jason A; Beigel, John H; Paolino, Kristopher M; Voell, Jocelyn; Castellano, Amy R; Hu, Zonghui; Muñoz, Paula; Moon, James E; Ruck, Richard C; Bennett, Jason W; Twomey, Patrick S; Gutiérrez, Ramiro L; Remich, Shon A; Hack, Holly R; Wisniewski, Meagan L; Josleyn, Matthew D; Kwilas, Steven A; Van Deusen, Nicole; Mbaya, Olivier Tshiani; Zhou, Yan; Stanley, Daphne A; Jing, Wang; Smith, Kirsten S; Shi, Meng; Ledgerwood, Julie E; Graham, Barney S; Sullivan, Nancy J; Jagodzinski, Linda L; Peel, Sheila A; Alimonti, Judie B; Hooper, Jay W; Silvera, Peter M; Martin, Brian K; Monath, Thomas P; Ramsey, W Jay; Link, Charles J; Lane, H Clifford; Michael, Nelson L; Davey, Richard T; Thomas, Stephen J

2017-01-26

The worst Ebola virus disease (EVD) outbreak in history has resulted in more than 28,000 cases and 11,000 deaths. We present the final results of two phase 1 trials of an attenuated, replication-competent, recombinant vesicular stomatitis virus (rVSV)-based vaccine candidate designed to prevent EVD. We conducted two phase 1, placebo-controlled, double-blind, dose-escalation trials of an rVSV-based vaccine candidate expressing the glycoprotein of a Zaire strain of Ebola virus (ZEBOV). A total of 39 adults at each site (78 participants in all) were consecutively enrolled into groups of 13. At each site, volunteers received one of three doses of the rVSV-ZEBOV vaccine (3 million plaque-forming units [PFU], 20 million PFU, or 100 million PFU) or placebo. Volunteers at one of the sites received a second dose at day 28. Safety and immunogenicity were assessed. The most common adverse events were injection-site pain, fatigue, myalgia, and headache. Transient rVSV viremia was noted in all the vaccine recipients after dose 1. The rates of adverse events and viremia were lower after the second dose than after the first dose. By day 28, all the vaccine recipients had seroconversion as assessed by an enzyme-linked immunosorbent assay (ELISA) against the glycoprotein of the ZEBOV-Kikwit strain. At day 28, geometric mean titers of antibodies against ZEBOV glycoprotein were higher in the groups that received 20 million PFU or 100 million PFU than in the group that received 3 million PFU, as assessed by ELISA and by pseudovirion neutralization assay. A second dose at 28 days after dose 1 significantly increased antibody titers at day 56, but the effect was diminished at 6 months. This Ebola vaccine candidate elicited anti-Ebola antibody responses. After vaccination, rVSV viremia occurred frequently but was transient. These results support further evaluation of the vaccine dose of 20 million PFU for preexposure prophylaxis and suggest that a second dose may boost antibody responses

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...... understanding of their role in the activation of macrophages and their stimulation of proinflammatory cytokine production in early viral infection was the main goal of this study. Our in vitro model study showed that probiotic bacteria, either from the species Lactobacillus or Bifidobacteria have the ability...... dehydrogenases activity could be implied as the first indicator of potential inhibitory effects of the probiotics on virus replication. The interactions between probiotic bacteria, macrophages and vesicular stomatitis virus (VSV), markedly depended on the bacterial strain studied....

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

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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.

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

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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.

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 V_cmax 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.

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

International Nuclear Information System (INIS)

Whitt, M.A.; Chong, L.; Rose, J.K.

1989-01-01

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

Infection of Melanoplus sanguinipes grasshoppers following ingestion of rangeland plant species harboring vesicular stomatitis virus.

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Drolet, Barbara S; Stuart, Melissa A; Derner, Justin D

2009-05-01

Knowledge of the many mechanisms of vesicular stomatitis virus (VSV) transmission is critical for understanding of the epidemiology of sporadic disease outbreaks in the western United States. Migratory grasshoppers [Melanoplus sanguinipes (Fabricius)] have been implicated as reservoirs and mechanical vectors of VSV. The grasshopper-cattle-grasshopper transmission cycle is based on the assumptions that (i) virus shed from clinically infected animals would contaminate pasture plants and remain infectious on plant surfaces and (ii) grasshoppers would become infected by eating the virus-contaminated plants. Our objectives were to determine the stability of VSV on common plant species of U.S. Northern Plains rangelands and to assess the potential of these plant species as a source of virus for grasshoppers. Fourteen plant species were exposed to VSV and assayed for infectious virus over time (0 to 24 h). The frequency of viable virus recovery at 24 h postexposure was as high as 73%. The two most common plant species in Northern Plains rangelands (western wheatgrass [Pascopyrum smithii] and needle and thread [Hesperostipa comata]) were fed to groups of grasshoppers. At 3 weeks postfeeding, the grasshopper infection rate was 44 to 50%. Exposure of VSV to a commonly used grasshopper pesticide resulted in complete viral inactivation. This is the first report demonstrating the stability of VSV on rangeland plant surfaces, and it suggests that a significant window of opportunity exists for grasshoppers to ingest VSV from contaminated plants. The use of grasshopper pesticides on pastures would decrease the incidence of a virus-amplifying mechanical vector and might also decontaminate pastures, thereby decreasing the inter- and intraherd spread of VSV.

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

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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; Cepko, Constance L

2015-08-01

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. © 2015 Wiley Periodicals, Inc.

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

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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.

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

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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.

Location of the binding domains for the RNA polymerase L and the ribonucleocapsid template within different halves of the NS phosphoprotein of vesicular stomatitis virus

International Nuclear Information System (INIS)

Emerson, S.U.; Schubert, M.

1987-01-01

Recombinant DNA techniques were used to delete regions of a cDNA clone of the phosphoprotein NS gene of vesicular stomatitis virus. The complete NS gene and four mutant genes containing internal or terminal deletions were inserted into a modified pGem4 vector under the transcriptional control of the page T7 promoter. Run-off transcripts were synthesized and translated in vitro to provide [ 35 S]methionine-labeled complete NS or deletion mutant NS proteins. Immune coprecipitation assays involving these proteins were developed to map the regions of the NS protein responsible for binding to the structural viral nucleocapsid protein N and the catalytic RNA polymerase protein L. The data indicate the NS protein is a bivalent protein consisting of two discrete functional domains. Contrary to previous suggestions, the negatively charged amino-terminal half of NS protein binds to L protein, while the carboxyl-terminal half of NS protein binds to both soluble recombinant nucleocapsid protein N and viral ribonucleocapsid template

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

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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

Subcellular distribution of swine vesicular disease virus proteins and alterations induced in infected cells: A comparative study with foot-and-mouth disease virus and vesicular stomatitis virus

International Nuclear Information System (INIS)

Martin-Acebes, Miguel A.; Gonzalez-Magaldi, Monica; Rosas, Maria F.; Borrego, Belen; Brocchi, Emiliana; Armas-Portela, Rosario; Sobrino, Francisco

2008-01-01

The intracellular distribution of swine vesicular disease virus (SVDV) proteins and the induced reorganization of endomembranes in IBRS-2 cells were analyzed. Fluorescence to new SVDV capsids appeared first upon infection, concentrated in perinuclear circular structures and colocalized to dsRNA. As in foot-and-mouth disease virus (FMDV)-infected cells, a vesicular pattern was predominantly found in later stages of SVDV capsid morphogenesis that colocalized with those of non-structural proteins 2C, 2BC and 3A. These results suggest that assembly of capsid proteins is associated to the replication complex. Confocal microscopy showed a decreased fluorescence to ER markers (calreticulin and protein disulfide isomerase), and disorganization of cis-Golgi gp74 and trans-Golgi caveolin-1 markers in SVDV- and FMDV-, but not in vesicular stomatitis virus (VSV)-infected cells. Electron microscopy of SVDV-infected cells at an early stage of infection revealed fragmented ER cisternae with expanded lumen and accumulation of large Golgi vesicles, suggesting alterations of vesicle traffic through Golgi compartments. At this early stage, FMDV induced different patterns of ER fragmentation and Golgi alterations. At later stages of SVDV cytopathology, cells showed a completely vacuolated cytoplasm containing vesicles of different sizes. Cell treatment with brefeldin A, which disrupts the Golgi complex, reduced SVDV (∼ 5 log) and VSV (∼ 4 log) titers, but did not affect FMDV growth. Thus, three viruses, which share target tissues and clinical signs in natural hosts, induce different intracellular effects in cultured cells

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.

Activation of Nrf2 Signaling Augments Vesicular Stomatitis Virus Oncolysis via Autophagy-Driven Suppression of Antiviral Immunity.

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Olagnier, David; Lababidi, Rassin R; Hadj, Samar Bel; Sze, Alexandre; Liu, Yiliu; Naidu, Sharadha Dayalan; Ferrari, Matteo; Jiang, Yuan; Chiang, Cindy; Beljanski, Vladimir; Goulet, Marie-Line; Knatko, Elena V; Dinkova-Kostova, Albena T; Hiscott, John; Lin, Rongtuan

2017-08-02

Oncolytic viruses (OVs) offer a promising therapeutic approach to treat multiple types of cancer. In this study, we show that the manipulation of the antioxidant network via transcription factor Nrf2 augments vesicular stomatitis virus Δ51 (VSVΔ51) replication and sensitizes cancer cells to viral oncolysis. Activation of Nrf2 signaling by the antioxidant compound sulforaphane (SFN) leads to enhanced VSVΔ51 spread in OV-resistant cancer cells and improves the therapeutic outcome in different murine syngeneic and xenograft tumor models. Chemoresistant A549 lung cancer cells that display constitutive dominant hyperactivation of Nrf2 signaling are particularly vulnerable to VSVΔ51 oncolysis. Mechanistically, enhanced Nrf2 signaling stimulated viral replication in cancer cells and disrupted the type I IFN response via increased autophagy. This study reveals a previously unappreciated role for Nrf2 in the regulation of autophagy and the innate antiviral response that complements the therapeutic potential of VSV-directed oncolysis against multiple types of OV-resistant or chemoresistant cancer. Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.

Modified vaccinia virus Ankara protects macaques against respiratory challenge with monkeypox virus.

NARCIS (Netherlands)

K.J. Stittelaar (Koert); G. van Amerongen (Geert); I. Kondova (Ivanela); R.F. van Lavieren (Rob); F.H. Pistoor (Frank); H.G.M. Niesters (Bert); G.J.J. van Doornum (Gerard); B.A.M. van der Zeijst (Ben); L. Mateo (Luis); P.J. Chaplin (Paul); A.D.M.E. Osterhaus (Albert); T. Kuiken (Thijs)

2005-01-01

textabstractThe use of classical smallpox vaccines based on vaccinia virus (VV) is associated with severe complications in both naive and immune individuals. Modified vaccinia virus Ankara (MVA), a highly attenuated replication-deficient strain of VV, has been proven to be safe in humans and

Curcumin and Boswellia serrata gum resin extract inhibit chikungunya and vesicular stomatitis virus infections in vitro.

Science.gov (United States)

von Rhein, Christine; Weidner, Tatjana; Henß, Lisa; Martin, Judith; Weber, Christopher; Sliva, Katja; Schnierle, Barbara S

2016-01-01

Chikungunya virus (CHIKV) is a mosquito-transmitted alphavirus that causes chikungunya fever and has infected millions of people mainly in developing countries. The associated disease is characterized by rash, high fever, and severe arthritis that can persist for years. CHIKV has adapted to Aedes albopictus, which also inhabits temperate regions including Europe and the United States of America. CHIKV has recently caused large outbreaks in Latin America. No treatment or licensed CHIKV vaccine exists. Traditional medicines are known to have anti-viral effects; therefore, we examined whether curcumin or Boswellia serrata gum resin extract have antiviral activity against CHIKV. Both compounds blocked entry of CHIKV Env-pseudotyped lentiviral vectors and inhibited CHIKV infection in vitro. In addition, vesicular stomatitis virus vector particles and viral infections were also inhibited to the same extent, indicating a broad antiviral activity. Although the bioavailability of these compounds is rather poor, they might be used as a lead structure to develop more effective antiviral drugs or might be used topically to prevent CHIKV spread in the skin after mosquito bites. Copyright © 2015 Elsevier B.V. All rights reserved.

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

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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.

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.

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.

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

2011-12-01

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.

Stomatal and Non-Stomatal Turbulent Deposition Flux of Ozone to a Managed Peatland

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Tarek S. El-Madany

2017-09-01

Full Text Available Ozone is a key trace gas in the troposphere; because it is a greenhouse gas, it is very reactive, and it is potentially toxic to humans, fauna, and vegetation. The main sink processes for ozone are chemical reactions and the turbulent deposition flux to the earth’s surface. The deposition process itself is rather complex: The interactions between co-varying drivers such as the tropospheric ozone concentration, turbulence, and chemical reactions are not well understood. In the case of ozone deposition to vegetation, another aspect that must be studied is the role of stomatal regulation for a wide range of conditions. Therefore, we measured turbulent deposition fluxes of ozone with the eddy covariance technique during the peak of the growing season in 2014 over a managed, rewetted peatland in NW Germany. The deposition flux was large during the day (up to −15 nmol m−2 s−1 and relatively small during the night (between −1 and −2 nmol m−2 s−1. Flux partitioning by applying the surface resistance analogy and further analysis showed that the stomatal uptake was smaller than non-stomatal deposition. The correction of stomatal conductance with the gross primary production (GPP improved the estimation of day- and nighttime stomatal deposition fluxes. Statistical analysis confirmed that the friction velocity (u* was the single most important driver of non-stomatal ozone deposition and that relationships with other environmental drivers are not linear and highly variable. Further research is needed to develop a better process understanding of non-stomatal ozone deposition, to quantify the role of surface deposition to the ozone budget of the atmospheric boundary layer, and to estimate uncertainties associated with the partitioning of ozone deposition into stomatal and non-stomatal fluxes.

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

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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.

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

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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 (P n ) primarily by promoting stomatal factors, resulting in increased relative growth rates and accelerated soybean seedling growth. High-dose BPA decreases the P n 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 P n by promoting stomatal factors while high-dose BPA decreased the P n 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.

A novel method for analysis of membrane microdomains: vesicular stomatitis virus glycoprotein microdomains change in size during infection, and those outside of budding sites resemble sites of virus budding

International Nuclear Information System (INIS)

Brown, Erica L.; Lyles, Douglas S.

2003-01-01

Membrane proteins, including viral envelope glycoproteins, may be organized into areas of locally high concentration, commonly referred to as membrane microdomains. Some viruses bud from detergent-resistant microdomains referred to as lipid rafts. However, vesicular stomatitis virus (VSV) serves as a prototype for viruses that bud from areas of plasma membrane that are not detergent resistant. We developed a new analytical method for immunoelectron microscopy data to determine whether the VSV envelope glycoprotein (G protein) is organized into plasma membrane microdomains. This method was used to quantify the distribution of the G protein in microdomains in areas of plasma membrane that did not contain budding sites. These microdomains were compared to budding virus envelopes to address the question of whether G protein-containing microdomains were formed only at the sites of budding. At early times postinfection, most of the G protein was organized into membrane microdomains outside of virus budding sites that were approximately 100-150 nm, with smaller amounts distributed into larger microdomains. In contrast to early times postinfection, the increased level of G protein in the host plasma membrane at later times postinfection led to distribution of G protein among membrane microdomains of a wider variety of sizes, rather than a higher G protein concentration in the 100- to 150-nm microdomains. VSV budding occurred in G protein-containing microdomains with a range of sizes, some of which were smaller than the virus envelope. These microdomains extended in size to a maximum of 300-400 nm from the tip of the budding virion. The data support a model for virus assembly in which G protein organizes into membrane microdomains that resemble virus envelopes prior to formation of budding sites, and these microdomains serve as the sites of assembly of internal virion components

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

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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.

CD4(+) T cell-mediated protection against a lethal outcome of systemic infection with vesicular stomatitis virus requires CD40 ligand expression, but not IFN-gamma or IL-4

DEFF Research Database (Denmark)

Andersen, C; Jensen, T; Nansen, A

1999-01-01

experiments using B cell- and T cell-deficient recipients revealed that no protection could be obtained in the absence of B cells, whereas treatment with virus-specific immune (IgG) serum controlled viral spreading to the central nervous system (CNS), but did not necessarily accomplish virus elimination......To investigate the mechanism(s) whereby T cells protect against a lethal outcome of systemic infection with vesicular stomatitis virus, mice with targeted defects in genes central to T cell function were tested for resistance to i.v. infection with this virus. Our results show that mice lacking...... the capacity to secrete both IFN-gamma and perforin completely resisted disease. Similar results were obtained using IL-4 knockout mice, indicating that neither cell-mediated nor T(h)2-dependent effector systems were required. In contrast, mice deficient in expression of CD40 ligand were more susceptible than...

Safety studies on intravenous administration of oncolytic recombinant vesicular stomatitis virus in purpose-bred beagle dogs.

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LeBlanc, Amy K; Naik, Shruthi; Galyon, Gina D; Jenks, Nathan; Steele, Mike; Peng, Kah-Whye; Federspiel, Mark J; Donnell, Robert; Russell, Stephen J

2013-12-01

VSV-IFNβ-NIS is a novel recombinant oncolytic vesicular stomatitis virus (VSV) with documented efficacy and safety in preclinical murine models of cancer. To facilitate clinical translation of this promising oncolytic therapy in patients with disseminated cancer, we are utilizing a comparative oncology approach to gather data describing the safety and efficacy of systemic VSV-IFNβ-NIS administration in dogs with naturally occurring cancer. In support of this, we executed a dose-escalation study in purpose-bred dogs to determine the maximum tolerated dose (MTD) of systemic VSV-hIFNβ-NIS, characterize the adverse event profile, and describe routes and duration of viral shedding in healthy, immune-competent dogs. The data indicate that an intravenous dose of 10(10) TCID50 is well tolerated in dogs. Expected adverse events were mild to moderate fever, self-limiting nausea and vomiting, lymphopenia, and oral mucosal lesions. Unexpected adverse events included prolongation of partial thromboplastin time, development of bacterial urinary tract infection, and scrotal dermatitis, and in one dog receiving 10(11) TCID50 (10 × the MTD), the development of severe hepatotoxicity and symptoms of shock leading to euthanasia. Viral shedding data indicate that detectable viral genome in blood diminishes rapidly with anti-VSV neutralizing antibodies detectable in blood as early as day 5 postintravenous virus administration. While low levels of viral genome copies were detectable in plasma, urine, and buccal swabs of dogs treated at the MTD, no infectious virus was detectable in plasma, urine, or buccal swabs at any of the doses tested. These studies confirm that VSV can be safely administered systemically in dogs, justifying the use of oncolytic VSV as a novel therapy for the treatment of canine cancer.

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

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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.

Pesti Des Petits ruminants virus infection in animals

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Chauhan H.C.

2009-08-01

Full Text Available For centuries morbillivirus infections have had a huge impact on both human beings and animals. Morbilliviruses are highly contagious pathogens that cause some of the most devastating viral diseases of humans and animals world wide. They include measles virus (MV, canine distemper virus (CDV, rinderpest virus (RPV and peste des petits ruminants (PPRV virus. Furthermore, new emerging infectious diseases of morbilliviruses with significant ecological consequences of marine mammals have been discovered in the past decades. Phocid distemper virus (PDV in seals and the cetacean morbillivirus (CMV have been found in dolphins, whales and porpoises. Peste des petits ruminants (PPR is a highly contagious ,infectious , an acute or sub acute viral disease of domestic and wild small ruminants characterized by fever, oculonasal discharges, stomatitis, conjunctivitis, gastroenteritis and pneumonia. Goats are more severely affected than sheep. It is also known as pseudorinderpest of small ruminants, pest of small ruminants, pest of sheep and goats, kata, stomatitis- pneumoentritis syndrome, contagious pustular stomatitis and pneumoentritis complex. It is one of the major notifiable diseases of the World Organization for Animal Health (OIE. [Vet. World 2009; 2(4.000: 150-155

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...... probes. The limits of detection ware found to be less than 10 50% tissue culture infective doses/ml for both serotypes. The diagnostic value of the new method was tested with clinical materials from experimentally infected pigs, and it is concluded that the method is a powerful tool for the rapid...

Immunogenicity in African Green Monkeys of M Protein Mutant Vesicular Stomatitis Virus Vectors and Contribution of Vector-Encoded Flagellin

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Marlena M. Westcott

2018-03-01

Full Text Available Recombinant vesicular stomatitis virus (VSV is a promising platform for vaccine development. M51R VSV, an attenuated, M protein mutant strain, is an effective inducer of Type I interferon and dendritic cell (DC maturation, which are desirable properties to exploit for vaccine design. We have previously evaluated M51R VSV (M51R and M51R VSV that produces flagellin (M51R-F as vaccine vectors using murine models, and found that flagellin enhanced DC activation and VSV-specific antibody production after low-dose vaccination. In this report, the immunogenicity of M51R vectors and the adjuvant effect of virus-produced flagellin were evaluated in nonhuman primates following high-dose (108 pfu and low-dose (105 pfu vaccination. A single intramuscular vaccination of African green monkeys with M51R or M51R-F induced VSV-specific, dose-dependent humoral immune responses. Flagellin induced a significant increase in antibody production (IgM, IgG and neutralizing antibody at the low vaccination dose. A VSV-specific cellular response was detected at 6 weeks post-vaccination, but was neither dose-dependent nor enhanced by flagellin; similar numbers of VSV-specific, IFNγ-producing cells were detected in lymph node and spleen of all animals. These results indicate that virus-directed, intracellular flagellin production may improve VSV-based vaccines encoding heterologous antigens by lowering the dose required to achieve humoral immunity.

Permissivity of the NCI-60 cancer cell lines to oncolytic Vaccinia Virus GLV-1h68

International Nuclear Information System (INIS)

Ascierto, Maria Libera; Bedognetti, Davide; Uccellini, Lorenzo; Rossano, Fabio; Ascierto, Paolo A; Stroncek, David F; Restifo, Nicholas P; Wang, Ena; Szalay, Aladar A; Marincola, Francesco M; Worschech, Andrea; Yu, Zhiya; Adams, Sharon; Reinboth, Jennifer; Chen, Nanhai G; Pos, Zoltan; Roychoudhuri, Rahul; Di Pasquale, Giovanni

2011-01-01

Oncolytic viral therapy represents an alternative therapeutic strategy for the treatment of cancer. We previously described GLV-1h68, a modified Vaccinia Virus with exclusive tropism for tumor cells, and we observed a cell line-specific relationship between the ability of GLV-1h68 to replicate in vitro and its ability to colonize and eliminate tumor in vivo. In the current study we surveyed the in vitro permissivity to GLV-1h68 replication of the NCI-60 panel of cell lines. Selected cell lines were also tested for permissivity to another Vaccinia Virus and a vesicular stomatitis virus (VSV) strain. In order to identify correlates of permissity to viral infection, we measured transcriptional profiles of the cell lines prior infection. We observed highly heterogeneous permissivity to VACV infection amongst the cell lines. The heterogeneity of permissivity was independent of tissue with the exception of B cell derivation. Cell lines were also tested for permissivity to another Vaccinia Virus and a vesicular stomatitis virus (VSV) strain and a significant correlation was found suggesting a common permissive phenotype. While no clear transcriptional pattern could be identified as predictor of permissivity to infection, some associations were observed suggesting multifactorial basis permissivity to viral infection. Our findings have implications for the design of oncolytic therapies for cancer and offer insights into the nature of permissivity of tumor cells to viral infection

Modelling stomatal ozone flux and deposition to grassland communities across Europe

International Nuclear Information System (INIS)

Ashmore, M.R.; Bueker, P.; Emberson, L.D.; Terry, A.C.; Toet, S.

2007-01-01

Regional scale modelling of both ozone deposition and the risk of ozone impacts is poorly developed for grassland communities. This paper presents new predictions of stomatal ozone flux to grasslands at five different locations in Europe, using a mechanistic model of canopy development for productive grasslands to generate time series of leaf area index and soil water potential as inputs to the stomatal component of the DO 3 SE ozone deposition model. The parameterisation of both models was based on Lolium perenne, a dominant species of productive pasture in Europe. The modelled seasonal time course of stomatal ozone flux to both the whole canopy and to upper leaves showed large differences between climatic zones, which depended on the timing of the start of the growing season, the effect of soil water potential, and the frequency of hay cuts. Values of modelled accumulated flux indices and the AOT40 index showed a five-fold difference between locations, but the locations with the highest flux differed depending on the index used; the period contributing to the accumulation of AOT40 did not always coincide with the modelled period of active ozone canopy uptake. Use of a fixed seasonal profile of leaf area index in the flux model produced very different estimates of annual accumulated total canopy and leaf ozone flux when compared with the flux model linked to a simulation of canopy growth. Regional scale model estimates of both the risks of ozone impacts and of total ozone deposition will be inaccurate unless the effects of climate and management in modifying grass canopy growth are incorporated. - Modelled stomatal flux of ozone to productive grasslands in Europe shows different spatial and temporal variation to AOT40, and is modified by management and soil water status

Inactivation by gamma irradiation of animal viruses in simulated laboratory effluent

International Nuclear Information System (INIS)

Thomas, F.C.; Ouwerkerk, T.; McKercher, P.

1982-01-01

Several animal viruses were treated with gamma radiation from a 60 Co source under conditions which might be found in effluent from an animal disease laboratory. Swine vesicular disease virus, vesicular stomatitis virus, and blue-tongue virus were irradiated in tissues from experimentally infected animals. Pseudorabies virus, fowl plague virus, swine vesicular disease virus, and vesicular stomatitis virus were irradiated in liquid animal feces. All were tested in animals and in vitro. The D 10 values, that is, the doses required to reduce infectivity by 1 log 10 , were not apparently different from those expected from predictions based on other data and theoretical considerations. The existence of the viruses in pieces of tissues or in liquid feces made no differences in the efficacy of the gamma radiation for inactivating them. Under the ''worst case'' conditions (most protective for virus) simulated in this study, no infectious agents would survive 4.0 Mrads

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.

Characterization of pH-sensitive molecular switches that trigger the structural transition of vesicular stomatitis virus glycoprotein from the postfusion state toward the prefusion state.

Science.gov (United States)

Ferlin, Anna; Raux, Hélène; Baquero, Eduard; Lepault, Jean; Gaudin, Yves

2014-11-01

Vesicular stomatitis virus (VSV; the prototype rhabdovirus) fusion is triggered at low pH and mediated by glycoprotein G, which undergoes a low-pH-induced structural transition. A unique feature of rhabdovirus G is that its conformational change is reversible. This allows G to recover its native prefusion state at the viral surface after its transport through the acidic Golgi compartments. The crystal structures of G pre- and postfusion states have been elucidated, leading to the identification of several acidic amino acid residues, clustered in the postfusion trimer, as potential pH-sensitive switches controlling the transition back toward the prefusion state. We mutated these residues and produced a panel of single and double mutants whose fusion properties, conformational change characteristics, and ability to pseudotype a virus lacking the glycoprotein gene were assayed. Some of these mutations were also introduced in the genome of recombinant viruses which were further characterized. We show that D268, located in the segment consisting of residues 264 to 273, which refolds into postfusion helix F during G structural transition, is the major pH sensor while D274, D395, and D393 have additional contributions. Furthermore, a single passage of recombinant virus bearing the mutation D268L (which was demonstrated to stabilize the G postfusion state) resulted in a pseudorevertant with a compensatory second mutation, L271P. This revealed that the propensity of the segment of residues 264 to 273 to refold into helix F has to be finely tuned since either an increase (mutation D268L alone) or a decrease (mutation L271P alone) of this propensity is detrimental to the virus. Vesicular stomatitis virus enters cells via endocytosis. Endosome acidification induces a structural transition of its unique glycoprotein (G), which mediates fusion between viral and endosomal membranes. G conformational change is reversible upon increases in pH. This allows G to recover its native

Epizootic vesicular stomatitis in Colorado, 1982: epidemiologic and entomologic studies.

Science.gov (United States)

Walton, T E; Webb, P A; Kramer, W L; Smith, G C; Davis, T; Holbrook, F R; Moore, C G; Schiefer, T J; Jones, R H; Janney, G C

1987-01-01

An epizootic of vesicular stomatitis (VS) caused by the New Jersey serotype of VS virus affected livestock and humans in 14 western states in 1982-1983. Epidemiological observations were made on at least 10% of the cattle in 4 dairy herds that were located in the vicinity of Grand Junction, Colorado. High rates of neutralizing antibody to the New Jersey serotype were seen in all cattle regardless of whether livestock in the dairy had clinical VS or a decrease in mild production. Antibody titers remained high in these cattle for as long as 2 years after the epizootic. No virus isolations were made from 32 humans with clinical signs compatible with viral disease. Entomological information was obtained during the epizootic from 23 premises in northwestern Colorado. Insect collections yielded 4 isolates from Culicoides spp. midges, 2 from C. variipennis, and 1 each from C. stellifer and C. (Selfia) spp. This is the first report of VS virus isolations from field-collected Culicoides.

Heat Shock Protein 70 Enhances Mucosal Immunity against Human Norovirus When Coexpressed from a Vesicular Stomatitis Virus Vector

Science.gov (United States)

Ma, Yuanmei; Duan, Yue; Wei, Yongwei; Liang, Xueya; Niewiesk, Stefan; Oglesbee, Michael

2014-01-01

ABSTRACT Human norovirus (NoV) accounts for 95% of nonbacterial gastroenteritis worldwide. Currently, there is no vaccine available to combat human NoV as it is not cultivable and lacks a small-animal model. Recently, we demonstrated that recombinant vesicular stomatitis virus (rVSV) expressing human NoV capsid protein (rVSV-VP1) induced strong immunities in mice (Y. Ma and J. Li, J. Virol. 85:2942–2952, 2011). To further improve the safety and efficacy of the vaccine candidate, heat shock protein 70 (HSP70) was inserted into the rVSV-VP1 backbone vector. A second construct was generated in which the firefly luciferase (Luc) gene was inserted in place of HSP70 as a control for the double insertion. The resultant recombinant viruses (rVSV-HSP70-VP1 and rVSV-Luc-VP1) were significantly more attenuated in cell culture and viral spread in mice than rVSV-VP1. At the inoculation dose of 1.0 × 106 PFU, rVSV-HSP70-VP1 triggered significantly higher vaginal IgA than rVSV-VP1 and significantly higher fecal and vaginal IgA responses than rVSV-Luc-VP1, although serum IgG and T cell responses were similar. At the inoculation dose of 5.0 × 106 PFU, rVSV-HSP70-VP1 stimulated significantly higher T cell, fecal, and vaginal IgA responses than rVSV-VP1. Fecal and vaginal IgA responses were also significantly increased when combined vaccination of rVSV-VP1 and rVSV-HSP70 was used. Collectively, these data indicate that (i) insertion of an additional gene (HSP70 or Luc) into the rVSV-VP1 backbone further attenuates the VSV-based vaccine in vitro and in vivo, thus improving the safety of the vaccine candidate, and (ii) HSP70 enhances the human NoV-specific mucosal and T cell immunities triggered by a VSV-based human NoV vaccine. IMPORTANCE Human norovirus (NoV) is responsible for more than 95% of acute nonbacterial gastroenteritis worldwide. Currently, there is no vaccine for this virus. Development of a live attenuated vaccine for human NoV has not been possible because it is

Immunogenicity and protective efficacy of recombinant Modified Vaccinia virus Ankara candidate vaccines delivering West Nile virus envelope antigens

NARCIS (Netherlands)

Volz, Asisa; Lim, Stephanie; Kaserer, Martina; Pijlman, Gorben P.

2016-01-01

West Nile virus (WNV) cycles between insects and wild birds, and is transmitted via mosquito vectors to horses and humans, potentially causing severe neuroinvasive disease. Modified Vaccinia virus Ankara (MVA) is an advanced viral vector for developing new recombinant vaccines against infectious

Chikungunya Virus Vaccines: Viral Vector-Based Approaches.

Science.gov (United States)

Ramsauer, Katrin; Tangy, Frédéric

2016-12-15

In 2013, a major chikungunya virus (CHIKV) epidemic reached the Americas. In the past 2 years, >1.7 million people have been infected. In light of the current epidemic, with millions of people in North and South America at risk, efforts to rapidly develop effective vaccines have increased. Here, we focus on CHIKV vaccines that use viral-vector technologies. This group of vaccine candidates shares an ability to potently induce humoral and cellular immune responses by use of highly attenuated and safe vaccine backbones. So far, well-described vectors such as modified vaccinia virus Ankara, complex adenovirus, vesicular stomatitis virus, alphavirus-based chimeras, and measles vaccine Schwarz strain (MV/Schw) have been described as potential vaccines. We summarize here the recent data on these experimental vaccines, with a focus on the preclinical and clinical activities on the MV/Schw-based candidate, which is the first CHIKV-vectored vaccine that has completed a clinical trial. © The Author 2016. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail journals.permissions@oup.com.

A recombinant canine distemper virus expressing a modified rabies virus glycoprotein induces immune responses in mice.

Science.gov (United States)

Li, Zhili; Wang, Jigui; Yuan, Daoli; Wang, Shuang; Sun, Jiazeng; Yi, Bao; Hou, Qiang; Mao, Yaping; Liu, Weiquan

2015-06-01

Canine distemper virus (CDV) and rabies virus (RV) are two important pathogens of the dog. CDV, a member of the morbillivirus genus, has shown promise as an expression vector. The glycoprotein from RV is a main contributor to protective immunity and capable of eliciting the production of virus-neutralizing antibodies. In this study, we recovered an attenuated strain of canine distemper virus and constructed a recombinant virus, rCDV-RV-G, expressing a modified (R333Q) rabies virus glycoprotein (RV-G) of RV Flury strain LEP. RV-G expression by the recombinant viruses was confirmed. Furthermore, G was proved to be incorporated into the surface of CDV particles. While replication of the recombinant virus was slightly reduced compared with the parental CDV, it stably expressed the RV-G over ten serial passages. Inoculation of mice induced specific neutralizing antibodies against both RV-G and CDV. Therefore, the rCDV-RV-G has the potential as a vaccine that may be used to control rabies virus infection in dogs and other animals.

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 (m 7 G) diphosphates efficiently accepted pRNA, resulting in the formation of DAPpppA, 7-deaza-GpppA, and m 7 GpppA (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

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.

Improved virus removal in ceramic depth filters modified with MgO.

Science.gov (United States)

Michen, Benjamin; Fritsch, Johannes; Aneziris, Christos; Graule, Thomas

2013-02-05

Ceramic filters, working on the depth filtration principle, are known to improve drinking water quality by removing human pathogenic microorganisms from contaminated water. However, these microfilters show no sufficient barrier for viruses having diameters down to 20 nm. Recently, it was shown that the addition of positively charged materials, for example, iron oxyhydroxide, can improve virus removal by adsorption mechanisms. In this work, we modified a common ceramic filter based on diatomaceous earth by introducing a novel virus adsorbent material, magnesium oxyhydroxide, into the filter matrix. Such filters showed an improved removal of about 4-log in regard to bacteriophages MS2 and PhiX174. This is explained with the electrostatic enhanced adsorption approach that is the favorable adsorption of negatively charged viruses onto positively charged patches in an otherwise negatively charged filter matrix. Furthermore, we provide theoretical evidence applying calculations according to Derjaguin-Landau-Verwey-Overbeek theory to strengthen our experimental results. However, modified filters showed a significant variance in virus removal efficiency over the course of long-term filtration experiments with virus removal increasing with filter operation time (or filter aging). This is explained by transformational changes of MgO in the filter upon contact with water. It also demonstrates that filter history is of great concern when filters working on the adsorption principles are evaluated in regard to their retention performance as their surface characteristics may alter with use.

The Ecophysiology Of A Pinus Ponderosa Ecosystem Exposed To High Tropospheric Ozone: Implications For Stomatal And Non-Stomatal Ozone Fluxes

Science.gov (United States)

Fares, S.; McKay, M.; Goldstein, A.

2008-12-01

Ecosystems remove ozone from the troposphere through both stomatal and non-stomatal deposition. The portion of ozone taken up through stomata has an oxidative effect causing damage. We used a multi-year dataset to assess the physiological controls over ozone deposition. Environmental parameters, CO2 and ozone fluxes were measured continuously from January 2001 to December 2006 above a ponderosa pine plantation near Blodgett Forest, Georgetown, California. We studied the dynamic of NEE (Net Ecosystem Exchange, -838 g C m-2 yr-1) and water evapotranspiration on an annual and daily basis. These processes are tightly coupled to stomatal aperture which also controlled ozone fluxes. High levels of ozone concentrations (~ 100 ppb) were observed during the spring-summer period, with corresponding high levels of ozone fluxes (~ 30 μmol m-2 h-1). During the summer season, a large portion of the total ozone flux was due to non-stomatal processes, and we propose that a plant physiological control, releasing BVOC (Biogenic Volatile Organic Compounds), is mainly responsible. We analyzed the correlations of common ozone exposure metrics based on accumulation of concentrations (AOT40 and SUM0) with ozone fluxes (total, stomatal and non-stomatal). Stomatal flux showed poorer correlation with ozone concentrations than non-stomatal flux during summer and fall seasons, which largely corresponded to the growing period. We therefore suggest that AOT40 and SUM0 are poor predictors of ozone damage and that a physiologically based metric would be more effective.

Influence of nonsystemic transmission on the epidemiology of insect borne arboviruses: a case study of vesicular stomatitis epidemiology in the western United States.

Science.gov (United States)

Lord, Cynthia C; Tabachnick, Walter J

2002-05-01

Nonsystemic transmission, where a pathogen is transmitted between infected and uninfected vectors without the vertebrate host becoming viremic, may provide an explanation for transmission in systems where the vertebrate hosts have been difficult to identify. This transmission pathway had been previously demonstrated for tick-borne viruses and bacteria, but the recent demonstration for Simulium and vesicular stomatitis virus is the first for a blood-feeding insect. The epidemiology of vesicular stomatitis viruses has been difficult to understand, and nonsystemic transmission may be important. We use mathematical formulations of the basic reproduction number, R(0), to compare systemic and nonsystemic transmission. The absence of a latent period before host infectiousness in nonsystemic transmission may allow a more rapid increase in prevalence in the biting flies early in the development of a new outbreak. Aggregation of flies between hosts and at favored feeding sites on hosts will be important, but further data on nonsystemic transmission as a function of space and time are required to fully assess this pathway. The data needed to compare the two pathways and their relative roles in virus epidemiology are discussed.

Positive and negative peptide signals control stomatal density.

Science.gov (United States)

Shimada, Tomoo; Sugano, Shigeo S; Hara-Nishimura, Ikuko

2011-06-01

The stoma is a micro valve found on aerial plant organs that promotes gas exchange between the atmosphere and the plant body. Each stoma is formed by a strict cell lineage during the early stages of leaf development. Molecular genetics research using the model plant Arabidopsis has revealed the genes involved in stomatal differentiation. Cysteine-rich secretory peptides of the EPIDERMAL PATTERNING FACTOR-LIKE (EPFL) family play crucial roles as extracellular signaling factors. Stomatal development is orchestrated by the positive factor STOMAGEN/EPFL9 and the negative factors EPF1, EPF2, and CHALLAH/EPFL6 in combination with multiple receptors. EPF1 and EPF2 are produced in the stomatal lineage cells of the epidermis, whereas STOMAGEN and CHALLAH are derived from the inner tissues. These findings highlight the complex cell-to-cell and intertissue communications that regulate stomatal development. To optimize gas exchange, particularly the balance between the uptake of carbon dioxide (CO(2)) and loss of water, plants control stomatal activity in response to environmental conditions. The CO(2) level and light intensity influence stomatal density. Plants sense environmental cues in mature leaves and adjust the stomatal density of newly forming leaves, indicating the involvement of long-distance systemic signaling. This review summarizes recent research progress in the peptide signaling of stomatal development and discusses the evolutionary model of the signaling machinery.

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.

Stomatal characteristics of Eucalyptus grandis clonal hybrids in ...

African Journals Online (AJOL)

This study describes the stomatal response occurring during water stress and subsequent recovery of three Eucalyptus grandis clonal hybrids. 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.

Separating active and passive influences on stomatal control of transpiration.

Science.gov (United States)

McAdam, Scott A M; Brodribb, Timothy J

2014-04-01

Motivated by studies suggesting that the stomata of ferns and lycophytes do not conform to the standard active abscisic acid (ABA) -mediated stomatal control model, we examined stomatal behavior in a conifer species (Metasequoia glyptostroboides) that is phylogenetically midway between the fern and angiosperm clades. Similar to ferns, daytime stomatal closure in response to moderate water stress seemed to be a passive hydraulic process in M. glyptostroboides immediately alleviated by rehydrating excised shoots. Only after prolonged exposure to more extreme water stress did active ABA-mediated stomatal closure become important, because foliar ABA production was triggered after leaf turgor loss. The influence of foliar ABA on stomatal conductance and stomatal aperture was highly predictable and additive with the passive hydraulic influence. M. glyptostroboides thus occupies a stomatal behavior type intermediate between the passively controlled ferns and the characteristic ABA-dependent stomatal closure described in angiosperm herbs. These results highlight the importance of considering phylogeny as a major determinant of stomatal behavior.

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.

5′-Phospho-RNA Acceptor Specificity of GDP Polyribonucleotidyltransferase of Vesicular Stomatitis Virus in mRNA Capping

Science.gov (United States)

Ogino, Minako

2017-01-01

ABSTRACT 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

Isolation of a virus with rhabdovirus morphology from a white-beaked dolphin (Lagenorhynchus albirostris).

NARCIS (Netherlands)

A.D.M.E. Osterhaus (Albert); H.W.J. Broeders; K.S. Teppema; T. Kuiken (Thijs); J.A. House; H.W. Vos (Helma); I.K.G. Visser (Ilona)

1993-01-01

textabstractA virus with rhabdovirus morphology which proved to be antigenically distinct from rabies virus and vesicular stomatitis virus was isolated from a dolphin that had beached on the Dutch coast. Neutralizing antibodies to this virus were found in several European marine mammal species.

The evolution of RNA viruses: A population genetics view

Science.gov (United States)

Moya, Andrés; Elena, Santiago F.; Bracho, Alma; Miralles, Rosario; Barrio, Eladio

2000-01-01

RNA viruses are excellent experimental models for studying evolution under the theoretical framework of population genetics. For a proper justification of this thesis we have introduced some properties of RNA viruses that are relevant for studying evolution. On the other hand, population genetics is a reductionistic theory of evolution. It does not consider or make simplistic assumptions on the transformation laws within and between genotypic and phenotypic spaces. However, such laws are minimized in the case of RNA viruses because the phenotypic space maps onto the genotypic space in a much more linear way than on higher DNA-based organisms. Under experimental conditions, we have tested the role of deleterious and beneficial mutations in the degree of adaptation of vesicular stomatitis virus (VSV), a nonsegmented virus of negative strand. We also have studied how effective population size, initial genetic variability in populations, and environmental heterogeneity shapes the impact of mutations in the evolution of vesicular stomatitis virus. Finally, in an integrative attempt, we discuss pros and cons of the quasispecies theory compared with classic population genetics models for haploid organisms to explain the evolution of RNA viruses. PMID:10860958

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.

Combining sap flow and eddy covariance approaches to derive stomatal and non-stomatal O3 fluxes in a forest stand

International Nuclear Information System (INIS)

Nunn, A.J.; Cieslik, S.; Metzger, U.; Wieser, G.; Matyssek, R.

2010-01-01

Stomatal O 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 3 flux was 33% of the total O 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 3 flux and reflected stomatal regulation rather than O 3 exposure, paralleling the daily courses of canopy conductance for water vapor and eddy covariance-based total stand-level O 3 flux. The demonstrated combination of sap flow and eddy covariance approaches supports the development of O 3 risk assessment in forests from O 3 exposure towards flux-based concepts. - Combined tree sap flow and eddy covariance-based methodologies yield stomatal O 3 flux as 33% in total stand flux.

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.

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...

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.

Separating Active and Passive Influences on Stomatal Control of Transpiration[OPEN

Science.gov (United States)

McAdam, Scott A.M.; Brodribb, Timothy J.

2014-01-01

Motivated by studies suggesting that the stomata of ferns and lycophytes do not conform to the standard active abscisic acid (ABA) -mediated stomatal control model, we examined stomatal behavior in a conifer species (Metasequoia glyptostroboides) that is phylogenetically midway between the fern and angiosperm clades. Similar to ferns, daytime stomatal closure in response to moderate water stress seemed to be a passive hydraulic process in M. glyptostroboides immediately alleviated by rehydrating excised shoots. Only after prolonged exposure to more extreme water stress did active ABA-mediated stomatal closure become important, because foliar ABA production was triggered after leaf turgor loss. The influence of foliar ABA on stomatal conductance and stomatal aperture was highly predictable and additive with the passive hydraulic influence. M. glyptostroboides thus occupies a stomatal behavior type intermediate between the passively controlled ferns and the characteristic ABA-dependent stomatal closure described in angiosperm herbs. These results highlight the importance of considering phylogeny as a major determinant of stomatal behavior. PMID:24488969

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.

Polyelectrolyte-modified cowpea mosaic virus for the synthesis of gold nanoparticles.

Science.gov (United States)

Aljabali, Alaa A A; Evans, David J

2014-01-01

Polyelectrolyte surface-modified cowpea mosaic virus (CPMV) can be used for the templated synthesis of narrowly dispersed gold nanoparticles. Cationic polyelectrolyte, poly(allylamine) hydrochloride, is electrostatically bound to the external surface of the virus capsid. The polyelectrolyte-coated CPMV promotes adsorption of aqueous gold hydroxide anionic species, prepared from gold(III) chloride and potassium carbonate, that are easily reduced to form CPMV-templated gold nanoparticles. The process is simple and environmentally benign using only water as solvent at ambient temperature.

Single-Domain Antibodies as Tools to Perturb and Study RNA Viruses

NARCIS (Netherlands)

Hanke, Leo

2017-01-01

In this thesis, I describe the generation and characterization of alpaca-derived, antiviral, single-domain antibody fragments (VHHs). The antiviral targets of the described VHHs are the nuclear proteins of influenza A virus (IAV) and vesicular stomatitis virus (VSV). The described VHHs protect cells

Stomatal uptake and stomatal deposition of ozone in isoprene and monoterpene emitting plants.

Science.gov (United States)

Fares, S; Loreto, F; Kleist, E; Wildt, J

2008-01-01

Volatile isoprenoids were reported to protect plants against ozone. To understand whether this could be the result of a direct scavenging of ozone by these molecules, the stomatal and non-stomatal uptake of ozone was estimated in plants emitting isoprene or monoterpenes. Ozone uptake by holm oak (Quercus ilex, a monoterpene emitter) and black poplar (Populus nigra, an isoprene emitter) was studied in whole plant enclosures (continuously stirred tank reactors, CSTR). The ozone uptake by plants was estimated measuring ozone concentration at the inlet and outlet of the reactors, after correcting for the uptake of the enclosure materials. Destruction of ozone at the cuticle or at the plant stems was found to be negligible compared to the ozone uptake through the stomata. For both plant species, a relationship between stomatal conductance and ozone uptake was found. For the poplar, the measured ozone losses were explained by the uptake of ozone through the stomata only, and ozone destruction by gas phase reactions with isoprene was negligible. For the oak, gas phase reactions of ozone with the monoterpenes emitted by the plants contributed significantly to ozone destruction. This was confirmed by two different experiments showing a) that in cases of high stomatal conductance but under low CO(2) concentration, a reduction of monoterpene emission was still associated with reduced O(3) uptake; and b) that ozone losses due to the gas phase reactions only can be measured when using the exhaust from a plant chamber to determine the gas phase reactivity in an empty reaction chamber. Monoterpenes can therefore relevantly scavenge ozone at leaf level contributing to protection against ozone.

Sleep and behavior during vesicular stomatitis virus induced encephalitis in BALB/cJ and C57BL/6J mice

Science.gov (United States)

Machida, Mayumi; Ambrozewicz, Marta A.; Breving, Kimberly; Wellman, Laurie L.; Yang, Linghui; Ciavarra, Richard P.; Sanford, Larry D.

2013-01-01

Intranasal application of vesicular stomatitis virus (VSV) produces a well-characterized model of viral encephalitis in mice. Within one day post-infection (PI), VSV travels to the olfactory bulb and, over the course of 7 days, it infects regions and tracts extending into the brainstem followed by clearance and recovery in most mice by PI day 14 (PI 14). Infectious diseases are commonly accompanied by excessive sleepiness; thus, sleep is considered a component of the acute phase response to infection. In this project, we studied the relationship between sleep and VSV infection using C57BL/6 (B6) and BALB/c mice. Mice were implanted with transmitters for recording EEG, activity and temperature by telemetry. After uninterrupted baseline recordings were collected for 2 days, each animal was infected intranasally with a single low dose of VSV (5 × 104 PFU). Sleep was recorded for 15 consecutive days and analyzed on PI 0, 1, 3, 5, 7, 10, and 14. Compared to baseline, amounts of non-rapid eye movement sleep (NREM) were increased in B6 mice during the dark period of PI 1–5, whereas rapid eye movement sleep (REM) was significantly reduced during the light periods of PI 0–14. In contrast, BALB/c mice showed significantly fewer changes in NREM and REM. These data demonstrate sleep architecture is differentially altered in these mouse strains and suggests that, in B6 mice, VSV can alter sleep before virus progresses into brain regions that control sleep. PMID:24055862

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.

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

Membrane fusion activity of vesicular stomatitis virus glycoprotein G is induced by low pH but not by heat or denaturant

International Nuclear Information System (INIS)

Yao Yi; Ghosh, Kakoli; Epand, Raquel F.; Epand, Richard M.; Ghosh, Hara P.

2003-01-01

The fusogenic envelope glycoprotein G of the rhabdovirus vesicular stomatitis virus (VSV) induces membrane fusion at acidic pH. At acidic pH the G protein undergoes a major structural reorganization leading to the fusogenic conformation. However, unlike other viral fusion proteins, the low-pH-induced conformational change of VSV G is completely reversible. As well, the presence of an α-helical coiled-coil motif required for fusion by a number of viral and cellular fusion proteins was not predicted in VSV G protein by using a number of algorithms. Results of pH dependence of the thermal stability of G protein as determined by intrinsic Trp fluorescence and circular dichroism (CD) spectroscopy show that the G protein is equally stable at neutral or acidic pH. Destabilization of G structure at neutral pH with either heat or urea did not induce membrane fusion or conformational change(s) leading to membrane fusion. Taken together, these data suggest that the mechanism of VSV G-induced fusion is distinct from the fusion mechanism of fusion proteins that involve a coiled-coil motif

A plasma membrane localization signal in the HIV-1 envelope cytoplasmic domain prevents localization at sites of vesicular stomatitis virus budding and incorporation into VSV virions.

Science.gov (United States)

Johnson, J E; Rodgers, W; Rose, J K

1998-11-25

Previous studies showed that the HIV-1 envelope (Env) protein was not incorporated into vesicular stomatitis virus (VSV) virions unless its cytoplasmic tail was replaced with that of the VSV glycoprotein (G). To determine whether the G tail provided a positive incorporation signal for Env, or if sequences in the Env tail prevented incorporation, we generated mutants of Env with its 150-amino-acid tail shortened to 29, 10, or 3 amino acids (Envtr mutants). Cells infected with VSV recombinants expressing these proteins or an Env-G tail hybrid showed similar amounts of Env protein at the surface. The Env-G tail hybrid or the Envtr3 mutant were incorporated at the highest levels into budding VSV virions. In contrast, the Envtr29 or Envtr10 mutants were incorporated poorly. These results defined a signal preventing incorporation within the 10 membrane-proximal amino acids of the Env tail. Confocal microscopy revealed that this signal functioned by causing localization of human immunodeficiency virus type 1 Env to plasma membrane domains distinct from the VSV budding sites, where VSV proteins were concentrated. Copyright 1998 Academic Press.

Efficient removal of pathogenic bacteria and viruses by multifunctional amine-modified magnetic nanoparticles.

Science.gov (United States)

Zhan, Sihui; Yang, Yang; Shen, Zhiqiang; Shan, Junjun; Li, Yi; Yang, Shanshan; Zhu, Dandan

2014-06-15

A novel amine-functionalized magnetic Fe3O4-SiO2-NH2 nanoparticle was prepared by layer-by-layer method and used for rapid removal of both pathogenic bacteria and viruses from water. The nanoparticles were characterized by TEM, EDS, XRD, XPS, FT-IR, BET surface analysis, magnetic property tests and zeta-potential measurements, respectively, which demonstrated its well-defined core-shell structures and strong magnetic responsivity. Pathogenic bacteria and viruses are often needed to be removed conveniently because of a lot of co-existing conditions. The amine-modified nanoparticles we prepared were attractive for capturing a wide range of pathogens including not only bacteriophage f2 and virus (Poliovirus-1), but also various bacteria such as S. aureus, E. coli O157:H7, P. aeruginosa, Salmonella, and B. subtilis. Using as-prepared amine-functionalized MNPs as absorbent, the nonspecific removal efficiency of E. coli O157:H7 or virus was more than 97.39%, while it is only 29.8% with Fe3O4-SiO2 particles. From joint removal test of bacteria and virus, there are over 95.03% harmful E. coli O157:H7 that can be removed from mixed solution with polyclonal anti-E. coli O157:H7 antibody modified nanoparticles. Moreover, the synergy effective mechanism has also been suggested. Copyright © 2014 Elsevier B.V. All rights reserved.

Interferon induction in bovine and feline monolayer cultures by four bluetongue virus serotypes.

OpenAIRE

Fulton, R W; Pearson, N J

1982-01-01

The interferon inducing ability of bluetongue viruses was studied in bovine and feline monolayer cultures inoculated with each of four bluetongue virus serotypes. Interferon was assayed by a plaque reduction method in monolayer cultures with vesicular stomatitis virus as challenge virus. Interferon was produced by bovine turbinate, Georgia bovine kidney, and Crandell feline kidney monolayer cultures in response to bluetongue virus serotypes 10, 11, 13 and 17. The antiviral substances produced...

Competitive virus assay method for titration of noncytopathogenic bovine viral diarrhea viruses (END⁺ and END⁻ viruses).

Science.gov (United States)

Muhsen, Mahmod; Ohi, Kota; Aoki, Hiroshi; Ikeda, Hidetoshi; Fukusho, Akio

2013-03-01

A new, reliable and secure virus assay method, named the competitive virus assay (CVA) method, has been established for the titration of bovine viral diarrhea viruses (BVDVs) that either show the exaltation of Newcastle disease virus (END) phenomenon or heterologous interference phenomenon (but not the END phenomenon). This method is based on the principle of (1) homologous interference between BVDVs, by using BVDV RK13/E(-) or BVDV RK13/E(+) strains as competitor virus, and (2) END phenomenon and heterologous interference, by using attenuated Newcastle disease virus (NDV) TCND strain as challenge virus. In titration of BVDV END(+) and BVDV END(-) viruses, no significant difference in estimated virus titer was observed between CVA and conventional methods. CVA method demonstrated comparable levels of sensitivity and accuracy as conventional END and interference methods, which require the use of a velogenic Miyadera strain of NDV and vesicular stomatitis virus (VSV), both of which are agents of high-risk diseases. As such, the CVA method is a safer alternative, with increased bio-safety and bio-containment, through avoidance of virulent strains that are commonly employed with conventional methods. Copyright © 2012 Elsevier B.V. All rights reserved.

Periodic fever, aphthous stomatitis, pharyngitis and cervical adenitis (pfapa) syndrome in children.

Science.gov (United States)

Semianchuk, Vira B

Periodic fever, aphthous stomatitis, pharyngitis and cervical adenitis (PFAPA) syndrome refers to a group of primary immunodeficiencies, namely autoinflammatory diseases. Most pediatricians and otolaryngologists do not suspect PFAPA syndrome when treating recurrent pharyngitis (according to Ukrainian classification - tonsillitis) and stomatitis. Therefore, patients with a given syndrome receive unnecessary treatment (antibiotic therapy or antiviral drugs) and the diagnosis is made late. The aim of the research was to provide pediatricians, family physicians and otolaryngologists with information on the importance of early diagnosis of PFAPA syndrome. The analysis of the prevalence and diagnosis of PFAPA syndrome in Ukraine and worldwide has been made as well as a late diagnosis of PFAPA syndrome in a child living in Ivano-Frankivsk, Ukraine has been described (case report). The Сase report 7-year-old boy, who grows and develops normally. The symptoms of pharyngitis including high body temperature (>40 º С), sore throat and white spots on the tonsils appeared for the first time at the age of two years. The boy received antibacterial drugs about 10 times a year. During a four-year period of recurrent episodes of the disease antimicrobial susceptibility testing to determine susceptibility of the oropharyngeal flora to the antibiotics were continuously performed, different blood tests for herpes viruses, Epstein-Barr virus infection and cytomegalovirus in particular were made using the enzyme immunoassay (EIA) and polymerase chain reaction (PCR) in addition to long-term treatment. An example of late diagnosing PFAPA syndrome (four years after the onset of first symptoms) resulting in regular examinations, medical manoeuvres, outpatient and inpatient treatment, use of antibiotic therapy including intravenous injections on a monthly basis has been studied.

Ecology of Candida-associated Denture Stomatitis

OpenAIRE

Budtz-Jørgensen, Ejvind

2011-01-01

Introduction of a prosthesis into the oral cavity results in profound alterations of the environmental conditions as the prosthesis and the underlying mucosa become colonized with oral microorganisms, including Candida spp. This may lead to denture stomatitis, a non-specific inflammatory reaction against microbial antigens, toxins and enzymes produced by the colonizing microorganisms. The role of Candida in the etiology of denture stomatitis is indicated by an increased number of yeasts on th...

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.

Middle east respiratory syndrome coronavirus spike protein delivered by modified vaccinia virus ankara efficiently induces virus-neutralizing antibodies

NARCIS (Netherlands)

F. Song (Fei); R. Fux (Robert); L.B.V. Provacia (Lisette); A. Volz (Asisa); M. Eickmann; S. Becker (Stephan); A.D.M.E. Osterhaus (Albert); B.L. Haagmans (Bart); G. Suttera (Gerd)

2013-01-01

textabstractMiddle East respiratory syndrome coronavirus (MERS-CoV) has recently emerged as a causative agent of severe respiratory disease in humans. Here, we constructed recombinant modified vaccinia virus Ankara (MVA) expressing full-length MERS-CoV spike (S) protein (MVA-MERS-S). The genetic

Virus-induced down-regulation of GmERA1A and GmERA1B genes enhances the stomatal response to abscisic acid and drought resistance in soybean.

Directory of Open Access Journals (Sweden)

Takuya Ogata

Full Text Available Drought is a major threat to global soybean production. The limited transformation potential and polyploid nature of soybean have hindered functional analysis of soybean genes. Previous research has implicated farnesylation in the plant's response to abscisic acid (ABA and drought tolerance. We therefore used virus-induced gene silencing (VIGS to evaluate farnesyltransferase genes, GmERA1A and GmERA1B (Glycine max Enhanced Response to ABA1-A and -B, as potential targets for increasing drought resistance in soybean. Apple latent spherical virus (ALSV-mediated GmERA1-down-regulated soybean leaves displayed an enhanced stomatal response to ABA and reduced water loss and wilting under dehydration conditions, suggesting that GmERA1A and GmERA1B negatively regulate ABA signaling in soybean guard cells. The findings provide evidence that the ALSV-VIGS system, which bypasses the need to generate transgenic plants, is a useful tool for analyzing gene function using only a single down-regulated leaf. Thus, the ALSV-VIGS system could constitute part of a next-generation molecular breeding pipeline to accelerate drought resistance breeding in soybean.

Multiplex RT-PCR and Automated Microarray for Detection of Eight Bovine Viruses.

Science.gov (United States)

Lung, O; Furukawa-Stoffer, T; Burton Hughes, K; Pasick, J; King, D P; Hodko, D

2017-12-01

Microarrays can be a useful tool for pathogen detection as it allow for simultaneous interrogation of the presence of a large number of genetic sequences in a sample. However, conventional microarrays require extensive manual handling and multiple pieces of equipment for printing probes, hybridization, washing and signal detection. In this study, a reverse transcription (RT)-PCR with an accompanying novel automated microarray for simultaneous detection of eight viruses that affect cattle [vesicular stomatitis virus (VSV), bovine viral diarrhoea virus type 1 and type 2, bovine herpesvirus 1, bluetongue virus, malignant catarrhal fever virus, rinderpest virus (RPV) and parapox viruses] is described. The assay accurately identified a panel of 37 strains of the target viruses and identified a mixed infection. No non-specific reactions were observed with a panel of 23 non-target viruses associated with livestock. Vesicular stomatitis virus was detected as early as 2 days post-inoculation in oral swabs from experimentally infected animals. The limit of detection of the microarray assay was as low as 1 TCID 50 /ml for RPV. The novel microarray platform automates the entire post-PCR steps of the assay and integrates electrophoretic-driven capture probe printing in a single user-friendly instrument that allows array layout and assay configuration to be user-customized on-site. © 2016 Her Majesty the Queen in Right of Canada.

Live Attenuated Recombinant Vaccine Protects Nonhuman Primates Against Ebola and Marburg Viruses

National Research Council Canada - National Science Library

Jones, Steven M; Feldmann, Heinz; Stroher, Ute; Geisbert, Joan B; Fernando, Lisa; Grolla, Allen; Klenk, Hans-Dieter; Sullivan, Nancy J; Volchkov, Viktor E; Fritz, Elizabeth A; Daddario, Kathleen M; Hensley, Lisa E; Jahrling, Peter B; Geisbert, Thomas W

2005-01-01

...). Here, we developed replication-competent vaccines against EBOV and MARV based on attenuated recombinant vesicular stomatitis virus vectors expressing either the EBOV glycoprotein or MARV glycoprotein...

Immunogenicity of a modified-live virus vaccine against bovine viral diarrhea virus types 1 and 2, infectious bovine rhinotracheitis virus, bovine parainfluenza-3 virus, and bovine respiratory syncytial virus when administered intranasally in young calves.

Science.gov (United States)

Xue, Wenzhi; Ellis, John; Mattick, Debra; Smith, Linda; Brady, Ryan; Trigo, Emilio

2010-05-14

The immunogenicity of an intranasally-administered modified-live virus (MLV) vaccine in 3-8 day old calves was evaluated against bovine viral diarrhea virus (BVDV) types 1 and 2, infectious bovine rhinotracheitis (IBR) virus, parainfluenza-3 (PI-3) virus and bovine respiratory syncytial virus (BRSV). Calves were intranasally vaccinated with a single dose of a multivalent MLV vaccine and were challenged with one of the respective viruses three to four weeks post-vaccination in five separate studies. There was significant sparing of diseases in calves intranasally vaccinated with the MLV vaccine, as indicated by significantly fewer clinical signs, lower rectal temperatures, reduced viral shedding, greater white blood cell and platelet counts, and less severe pulmonary lesions than control animals. This was the first MLV combination vaccine to demonstrate efficacy against BVDV types 1 and 2, IBR, PI-3 and BRSV in calves 3-8 days of age. Copyright 2010 Elsevier Ltd. All rights reserved.

Expected Net Benefit of Vaccinating Rangeland Sheep against Bluetongue Virus Using a Modified-Live versus Killed Virus Vaccine.

Science.gov (United States)

Munsick, Tristram R; Peck, Dannele E; Ritten, John P; Jones, Randall; Jones, Michelle; Miller, Myrna M

2017-01-01

Recurring outbreaks of bluetongue virus in domestic sheep of the US Intermountain West have prompted questions about the economic benefits and costs of vaccinating individual flocks against bluetongue (BT) disease. We estimate the cost of a BT outbreak on a representative rangeland sheep operation in the Big Horn Basin of the state of Wyoming using enterprise budgets and stochastic simulation. The latter accounts for variability in disease severity and lamb price, as well as uncertainty about when an outbreak will occur. We then estimate the cost of purchasing and administering a BT vaccine. Finally, we calculate expected annual net benefit of vaccinating under various outbreak intervals. Expected annual net benefit is calculated for both a killed virus (KV) vaccine and modified-live virus vaccine, using an observed price of $0.32 per dose for modified-live and an estimated price of $1.20 per dose for KV. The modified-live vaccine's expected annual net benefit has a 100% chance of being positive for an outbreak interval of 5, 10, or 20 years, and a 77% chance of being positive for a 50-year interval. The KV vaccine's expected annual net benefit has a 97% chance of being positive for a 5-year outbreak interval, and a 42% chance of being positive for a 10-year interval. A KV vaccine is, therefore, unlikely to be economically attractive to producers in areas exposed less frequently to BT disease. A modified-live vaccine, however, requires rigorous authorization before legal use can occur in Wyoming. To date, no company has requested to manufacture a modified-live vaccine for commercial use in Wyoming. The KV vaccine poses less risk to sheep reproduction and less risk of unintentional spread, both of which facilitate approval for commercial production. Yet, our results show an economically consequential tradeoff between a KV vaccine's relative safety and higher cost. Unless the purchase price is reduced below our assumed $1.20 per dose, producer adoption of a KV

mechanisms of drought resistance in grain ii:.stomatal regulation

African Journals Online (AJOL)

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STOMATAL REGULATION AND ROOT GROWTH ... maintenance of high plant water potential in common bean under stress was the function of stomatal regulation and/or root ... disadvantage since it will reduce CO2 fixation and hence may ...

Evaluation of a genetically modified foot-and-mouth disease virus vaccine candidate generated by reverse genetics

Science.gov (United States)

2012-01-01

Background Foot-and-mouth disease (FMD) is the most economically important and highly contagious disease of cloven-hoofed animals worldwide. Control of the disease has been mainly based on large-scale vaccinations with whole-virus inactivated vaccines. In recent years, a series of outbreaks of type O FMD occurred in China (including Chinese Taipei, Chinese Hong Kong) posed a tremendous threat to Chinese animal husbandry. Its causative agent, type O FMDV, has evolved into three topotypes (East–South Asia (ME-SA), Southeast Asia (SEA), Cathay (CHY)) in these regions, which represents an important obstacle to disease control. The available FMD vaccine in China shows generally good protection against ME-SA and SEA topotype viruses infection, but affords insufficient protection against some variants of the CHY topotype. Therefore, the choice of a new vaccine strain is of fundamental importance. Results The present study describes the generation of a full-length infectious cDNA clone of FMDV vaccine strain and a genetically modified virus with some amino acid substitutions in antigenic sites 1, 3, and 4, based on the established infectious clone. The recombinant viruses had similar growth properties to the wild O/HN/CHA/93 virus. All swine immunized with inactivated vaccine prepared from the O/HN/CHA/93 were fully protected from challenge with the viruses of ME-SA and SEA topotypes and partially protected against challenge with the virus of CHY topotype at 28 days post-immunization. In contrast, the swine inoculated with the genetically modified vaccine were completely protected from the infection of viruses of the three topotypes. Conclusions Some amino acid substitutions in the FMDV vaccine strain genome did not have an effect on the ability of viral replication in vitro. The vaccine prepared from genetically modified FMDV by reverse genetics significantly improved the protective efficacy to the variant of the CHY topotype, compared with the wild O/HN/CHA/93 virus

Evaluation of a genetically modified foot-and-mouth disease virus vaccine candidate generated by reverse genetics

Directory of Open Access Journals (Sweden)

Li Pinghua

2012-05-01

Full Text Available Abstract Background Foot-and-mouth disease (FMD is the most economically important and highly contagious disease of cloven-hoofed animals worldwide. Control of the disease has been mainly based on large-scale vaccinations with whole-virus inactivated vaccines. In recent years, a series of outbreaks of type O FMD occurred in China (including Chinese Taipei, Chinese Hong Kong posed a tremendous threat to Chinese animal husbandry. Its causative agent, type O FMDV, has evolved into three topotypes (East–South Asia (ME-SA, Southeast Asia (SEA, Cathay (CHY in these regions, which represents an important obstacle to disease control. The available FMD vaccine in China shows generally good protection against ME-SA and SEA topotype viruses infection, but affords insufficient protection against some variants of the CHY topotype. Therefore, the choice of a new vaccine strain is of fundamental importance. Results The present study describes the generation of a full-length infectious cDNA clone of FMDV vaccine strain and a genetically modified virus with some amino acid substitutions in antigenic sites 1, 3, and 4, based on the established infectious clone. The recombinant viruses had similar growth properties to the wild O/HN/CHA/93 virus. All swine immunized with inactivated vaccine prepared from the O/HN/CHA/93 were fully protected from challenge with the viruses of ME-SA and SEA topotypes and partially protected against challenge with the virus of CHY topotype at 28 days post-immunization. In contrast, the swine inoculated with the genetically modified vaccine were completely protected from the infection of viruses of the three topotypes. Conclusions Some amino acid substitutions in the FMDV vaccine strain genome did not have an effect on the ability of viral replication in vitro. The vaccine prepared from genetically modified FMDV by reverse genetics significantly improved the protective efficacy to the variant of the CHY topotype, compared with the

Matrix protein of vesicular stomatitis virus: a potent inhibitor of vascular endothelial growth factor and malignant ascites formation.

Science.gov (United States)

Zhou, Y; Wen, F; Zhang, P; Tang, R; Li, Q

2013-03-01

Malignant ascites is common in various types of cancers and is difficult to manage. Vascular endothelial growth factor (VEGF) has a pivotal role in malignant ascites. The matrix protein of vesicular stomatitis virus (VSVMP) has been shown to inhibit host gene expression and induce the apoptosis of cancer cells. The present study was designed to determine whether VSVMP suppresses the formation of ascites in ascites-producing peritoneal carcinomatosis. BALB/c female mice, 6-8 weeks old, bearing peritoneal tumors of H22 or MethA cells received an intraperitoneal administration of 50 μg VSVMP/250 μg liposome complexes, 50 μg empty plasmid/250 μg liposome complexes or 0.9% NaCl solution, respectively, every 2 days for 3 weeks. Administration of VSVMP resulted in a significant inhibition in ascites formation, improvement in health condition and prolonged survival of the treated mice. Decreased peritoneum osmolarity and reduced tumor vascularity coincided with dramatic reductions in the VEGF level in ascites fluid and plasma. Examination of floating tumor cells collected from the peritoneal wash revealed an apparently increased number of apoptotic cells and profound downregulation of VEGF mRNA in the VSVMP-treated mice. Our data indicate for the first time that in BALB/c mice bearing H22 or MethA cell peritoneal tumors, VSVMP may inhibit VEGF production and suppress angiogenesis, consequently abolishing ascites formation.

Mix-and-match: ligand-receptor pairs in stomatal development and beyond.

Science.gov (United States)

Torii, Keiko U

2012-12-01

Stomata are small valves on the plant epidermis balancing gas exchange and water loss. Stomata are formed according to positional cues. In Arabidopsis, two EPIDERMAL PATTERNING FACTOR (EPF) peptides, EPF1 and EPF2, are secreted from stomatal precursors enforcing proper stomatal patterning. Here, I review recent studies revealing the ligand-receptor pairs and revising the previously predicted relations between receptors specifying stomatal patterning: ERECTA-family and TOO MANY MOUTHS (TMM). Furthermore, EPF-LIKE9 (EPFL9/Stomagen) promotes stomatal differentiation from internal tissues. Two EPFL peptides specify inflorescence architecture, a process beyond stomatal development, as ligands for ERECTA. Thus, broadly expressed receptor kinases may regulate multiple developmental processes through perceiving different peptide ligands, each with a specialized expression pattern. TMM in the epidermis may fine-tune multiple EPF/EPFL signals to prevent signal interference. Copyright © 2012 Elsevier Ltd. All rights reserved.

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.

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.

Cross-Protection against Marburg Virus Strains by Using a Live, Attenuated Recombinant Vaccine

National Research Council Canada - National Science Library

Daddario-DiCaprio, Kathleen M; Geisbert, Thomas W; Geisbert, Joan B; Stroeher, Ute; Hensley, Lisa E; Grolla, Allen; Fritz, Elizabeth A; Feldmann, Friederike; Feldmann, Heinz; Jones, Steven M

2006-01-01

.... MARV is also considered to have potential as a biological weapon. Recently, we reported the development of a promising attenuated, replication-competent vaccine against MARV based on recombinant vesicular stomatitis virus (VSV...

Reconstructing Atmospheric CO2 Through The Paleocene-Eocene Thermal Maximum Using Stomatal Index and Stomatal Density Values From Ginkgo adiantoides

Science.gov (United States)

Barclay, R. S.; Wing, S. L.

2013-12-01

The Paleocene-Eocene Thermal Maximum (PETM) was a geologically brief interval of intense global warming 56 million years ago. It is arguably the best geological analog for a worst-case scenario of anthropogenic carbon emissions. The PETM is marked by a ~4-6‰ negative carbon isotope excursion (CIE) and extensive marine carbonate dissolution, which together are powerful evidence for a massive addition of carbon to the oceans and atmosphere. In spite of broad agreement that the PETM reflects a large carbon cycle perturbation, atmospheric concentrations of CO2 (pCO2) during the event are not well constrained. The goal of this study is to produce a high resolution reconstruction of pCO2 using stomatal frequency proxies (both stomatal index and stomatal density) before, during, and after the PETM. These proxies rely upon a genetically controlled mechanism whereby plants decrease the proportion of gas-exchange pores (stomata) in response to increased pCO2. Terrestrial sections in the Bighorn Basin, Wyoming, contain macrofossil plants with cuticle immediately bracketing the PETM, as well as dispersed plant cuticle from within the body of the CIE. These fossils allow for the first stomatal-based reconstruction of pCO2 near the Paleocene-Eocene boundary; we also use them to determine the relative timing of pCO2 change in relation to the CIE that defines the PETM. Preliminary results come from macrofossil specimens of Ginkgo adiantoides, collected from an ~200ka interval prior to the onset of the CIE (~230-30ka before), and just after the 'recovery interval' of the CIE. Stomatal index values decreased by 37% within an ~70ka time interval at least 100ka prior to the onset of the CIE. The decrease in stomatal index is interpreted as a significant increase in pCO2, and has a magnitude equivalent to the entire range of stomatal index adjustment observed in modern Ginkgo biloba during the anthropogenic CO2 rise during the last 150 years. The inferred CO2 increase prior to the

Recent advances in the development of vaccines for Ebola virus disease.

Science.gov (United States)

Ohimain, Elijah Ige

2016-01-04

Ebola virus is one of the most dangerous microorganisms in the world causing hemorrhagic fevers in humans and non-human primates. Ebola virus (EBOV) is a zoonotic infection, which emerges and re-emerges in human populations. The 2014 outbreak was caused by the Zaire strain, which has a kill rate of up to 90%, though 40% was recorded in the current outbreak. The 2014 outbreak is larger than all 20 outbreaks that have occurred since 1976, when the virus was first discovered. It is the first time that the virus was sustained in urban centers and spread beyond Africa into Europe and USA. Thus far, over 22,000 cases have been reported with about 50% mortality in one year. There are currently no approved therapeutics and preventive vaccines against Ebola virus disease (EVD). Responding to the devastating effe1cts of the 2014 outbreak and the potential risk of global spread, has spurred research for the development of therapeutics and vaccines. This review is therefore aimed at presenting the progress of vaccine development. Results showed that conventional inactivated vaccines produced from EBOV by heat, formalin or gamma irradiation appear to be ineffective. However, novel vaccines production techniques have emerged leading to the production of candidate vaccines that have been demonstrated to be effective in preclinical trials using small animal and non-human primates (NHP) models. Some of the promising vaccines have undergone phase 1 clinical trials, which demonstrated their safety and immunogenicity. Many of the candidate vaccines are vector based such as Vesicular Stomatitis Virus (VSV), Rabies Virus (RABV), Adenovirus (Ad), Modified Vaccinia Ankara (MVA), Cytomegalovirus (CMV), human parainfluenza virus type 3 (HPIV3) and Venezuelan Equine Encephalitis Virus (VEEV). Other platforms include virus like particle (VLP), DNA and subunit vaccines. Copyright © 2015 Elsevier B.V. All rights reserved.

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.

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.

Improving stomatal functioning at elevated growth air humidity: A review.

Science.gov (United States)

Fanourakis, Dimitrios; Bouranis, Dimitrios; Giday, Habtamu; Carvalho, Dália R A; Rezaei Nejad, Abdolhossein; Ottosen, Carl-Otto

2016-12-01

Plants grown at high relative air humidity (RH≥85%) are prone to lethal wilting upon transfer to conditions of high evaporative demand. The reduced survival of these plants is related to (i) increased cuticular permeability, (ii) changed anatomical features (i.e., longer pore length and higher stomatal density), (iii) reduced rehydration ability, (iv) impaired water potential sensitivity to leaf dehydration and, most importantly, (v) compromised stomatal closing ability. This review presents a critical analysis of the strategies which stimulate stomatal functioning during plant development at high RH. These include (a) breeding for tolerant cultivars, (b) interventions with respect to the belowground environment (i.e., water deficit, increased salinity, nutrient culture and grafting) as well as (c) manipulation of the aerial environment [i.e., increased proportion of blue light, increased air movement, temporal temperature rise, and spraying with abscisic acid (ABA)]. Root hypoxia, mechanical disturbance, as well as spraying with compounds mimicking ABA, lessening its inactivation or stimulating its within-leaf redistribution are also expected to improve stomatal functioning of leaves expanded in humid air. Available evidence leaves little doubt that genotypic and phenotypic differences in stomatal functioning following cultivation at high RH are realized through the intermediacy of ABA. Copyright © 2016 Elsevier GmbH. All rights reserved.

Plant twitter: ligands under 140 amino acids enforcing stomatal patterning.

Science.gov (United States)

Rychel, Amanda L; Peterson, Kylee M; Torii, Keiko U

2010-05-01

Stomata are an essential land plant innovation whose patterning and density are under genetic and environmental control. Recently, several putative ligands have been discovered that influence stomatal density, and they all belong to the epidermal patterning factor-like family of secreted cysteine-rich peptides. Two of these putative ligands, EPF1 and EPF2, are expressed exclusively in the stomatal lineage cells and negatively regulate stomatal density. A third, EPFL6 or CHALLAH, is also a negative regulator of density, but is expressed subepidermally in the hypocotyl. A fourth, EPFL9 or STOMAGEN, is expressed in the mesophyll tissues and is a positive regulator of density. Genetic evidence suggests that these ligands may compete for the same receptor complex. Proper stomatal patterning is likely to be an intricate process involving ligand competition, regional specificity, and communication between tissue layers. EPFL-family genes exist in the moss Physcomitrella patens, the lycophyte Selaginella moellendorffii, and rice, Oryza sativa, and their sequence analysis yields several genes some of which are related to EPF1, EPF2, EPFL6, and EPFL9. Presence of these EPFL family members in the basal land plants suggests an exciting hypothesis that the genetic components for stomatal patterning originated early in land plant evolution.

Natural variation in stomatal response to closing stimuli among Arabidopsis thaliana accessions after exposure to low VPD as a tool to recognize the mechanism of disturbed stomatal functioning.

Science.gov (United States)

Aliniaeifard, Sasan; van Meeteren, Uulke

2014-12-01

Stomatal responses to closing stimuli are disturbed after long-term exposure of plants to low vapour pressure deficit (VPD). The mechanism behind this disturbance is not fully understood. Genetic variation between naturally occurring ecotypes can be helpful to elucidate the mechanism controlling stomatal movements in different environments. We characterized the stomatal responses of 41 natural accessions of Arabidopsis thaliana to closing stimuli (ABA and desiccation) after they had been exposed for 4 days to moderate VPD (1.17 kPa) or low VPD (0.23 kPa). A fast screening system was used to test stomatal response to ABA using chlorophyll fluorescence imaging under low O2 concentrations of leaf discs floating on ABA solutions. In all accessions stomatal conductance (gs) was increased after prior exposure to low VPD. After exposure to low VPD, stomata of 39 out of 41 of the accessions showed a diminished ABA closing response; only stomata of low VPD-exposed Map-42 and C24 were as responsive to ABA as moderate VPD-exposed plants. In response to desiccation, most of the accessions showed a normal stomata closing response following low VPD exposure. Only low VPD-exposed Cvi-0 and Rrs-7 showed significantly less stomatal closure compared with moderate VPD-exposed plants. Using principle component analysis (PCA), accessions could be categorized to very sensitive, moderately sensitive, and less sensitive to closing stimuli. In conclusion, we present evidence for different stomatal responses to closing stimuli after long-term exposure to low VPD across Arabidopsis accessions. The variation can be a useful tool for finding the mechanism of stomatal malfunctioning. © The Author 2014. Published by Oxford University Press on behalf of the Society for Experimental Biology.

Salicaceae Endophytes Modulate Stomatal Behavior and Increase Water Use Efficiency in Rice

Directory of Open Access Journals (Sweden)

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.

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

[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.

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

Vesicular stomatitis virus modified with single chain IL-23 exhibits oncolytic activity against tumor cells in vitro and in vivo

OpenAIRE

Reiss, Carol Shoshkes

2010-01-01

James M Miller1, Sarah McNulty Bidula1,5, Troels Mygind Jensen1,6, Carol Shoshkes Reiss1,2,3,41Department of Biology, New York University, New York, NY, USA; 2Center for Neural Science, NYU; 3NYU Cancer Institute; 4Departments of Microbiology, NYU School of Medicine and Mt Sinai School of Medicine, New York, NY, USA; 5Present address Graduate Program, University of Pittsburgh School of Medicine, Pittsburgh, PA,USA 6Present address: Univercity of Copenhagen, Copenhagen, DenmarkAbstract: Viruse...

Infection of Melanoplus sanguinipes Grasshoppers following Ingestion of Rangeland Plant Species Harboring Vesicular Stomatitis Virus▿

Science.gov (United States)

Drolet, Barbara S.; Stuart, Melissa A.; Derner, Justin D.

2009-01-01

Knowledge of the many mechanisms of vesicular stomatitis virus (VSV) transmission is critical for understanding of the epidemiology of sporadic disease outbreaks in the western United States. Migratory grasshoppers [Melanoplus sanguinipes (Fabricius)] have been implicated as reservoirs and mechanical vectors of VSV. The grasshopper-cattle-grasshopper transmission cycle is based on the assumptions that (i) virus shed from clinically infected animals would contaminate pasture plants and remain infectious on plant surfaces and (ii) grasshoppers would become infected by eating the virus-contaminated plants. Our objectives were to determine the stability of VSV on common plant species of U.S. Northern Plains rangelands and to assess the potential of these plant species as a source of virus for grasshoppers. Fourteen plant species were exposed to VSV and assayed for infectious virus over time (0 to 24 h). The frequency of viable virus recovery at 24 h postexposure was as high as 73%. The two most common plant species in Northern Plains rangelands (western wheatgrass [Pascopyrum smithii] and needle and thread [Hesperostipa comata]) were fed to groups of grasshoppers. At 3 weeks postfeeding, the grasshopper infection rate was 44 to 50%. Exposure of VSV to a commonly used grasshopper pesticide resulted in complete viral inactivation. This is the first report demonstrating the stability of VSV on rangeland plant surfaces, and it suggests that a significant window of opportunity exists for grasshoppers to ingest VSV from contaminated plants. The use of grasshopper pesticides on pastures would decrease the incidence of a virus-amplifying mechanical vector and might also decontaminate pastures, thereby decreasing the inter- and intraherd spread of VSV. PMID:19286779

Detection of antibodies to transmissible gastroenteritis virus of swine by modified autoradiographic test

Energy Technology Data Exchange (ETDEWEB)

Stepanek, J; Hampl, J; Franz, J; Mensik, P; Skrobak, F [Vyzkumny Ustav Veterinarniho Lekarstvi, Brno-Medlanky (Czechoslovakia)

1982-08-01

A modified method of autoradiographic determination of virus antibodies of gastroenteritis of swine was developed. It is based on the actual reaction between antigen bound in cells of the infected cell cultures and antibodies in tested sera, which is visualized by rabbit antibodies labelled with /sup 125/I (/sup 125/I RaSw IgG antibody) to porcine IgG, on a sensitive radiograph and evaluated by darkening at the point of positive immunological reaction. Specificity of the test and mutual comparability and reproducibility of the results were confirmed by examining the known positive and negative sera and by a comparison with the results of the virus-neutralization test. Of the 36 examined porcine blood sera, antibodies were only proved autoradiographically in the samples positive also by virus-neutralization. In experimentally infected pigs, the same dynamics of antibody production was recorded by the two tests. They were, however, demonstrated autoradiographically the eighth day after infection, while by virus neutralization test as late as 14th day. Their level increased gradually till 35th day after infection.

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

Single-Injection Trivalent Filovirus 428 Vaccine: Proof of Concept Study in Outbred Guinea Pigs . J Infect Dis. 429 29. Murin, C. D., M. L. Fusco, Z...Jahrling, and J. F. Smith. 2000. Recombinant RNA replicons derived from attenuated 442 Venezuelan equine encephalitis virus protect guinea pigs and...platform, 65 including ease of production and characterization, absence of virus replication concerns and the 66 robust immune stimulatory activity

Distinction between infections with European and American/vaccine type PRRS virus after vaccination with a modified-live PRRS virus vaccine

DEFF Research Database (Denmark)

Bøtner, Anette; Strandbygaard, Bertel; Sørensen, K. J.

2000-01-01

In July 1996 a modified live Porcine reproductive and respiratory syndrome (PRRS) vaccine, based on an American (US) strain of the PRRS virus (PRRSV), was licensed in Denmark. The vaccine was licensed for use in 3-18 week old pigs, exclusively. Starting during the middle of October 1996, several...... herds who had recently begun vaccination, experienced acute PRRS-like symptoms including an increasing number of abortions and stillborn piglets and an increasing mortality in the nursing period. During the period from October 1996 until May 1997, the PRRS virus (PRRSV), identified as the vaccine....../US type of PRRSV, was isolated from fetuses, dead piglets, pleural fluids and/or lung tissues from 114 of such herds. These findings indicated the spread of the vaccine virus to non-vaccinated sows followed by transplacental infection of fetuses. Also, a number of not previously PRRSV infected and non...

Assessing urban habitat quality based on specific leaf area and stomatal characteristics of Plantago lanceolata L

International Nuclear Information System (INIS)

Kardel, F.; Wuyts, K.; Babanezhad, M.; Vitharana, U.W.A.; Wuytack, T.; Potters, G.; Samson, R.

2010-01-01

This study has evaluated urban habitat quality by studying specific leaf area (SLA) and stomatal characteristics of the common herb Plantago lanceolata L. SLA and stomatal density, pore surface and resistance were measured at 169 locations in the city of Gent (Belgium), distributed over four land use classes, i.e., sub-urban green, urban green, urban and industry. SLA and stomatal density significantly increased from sub-urban green towards more urbanised land use classes, while the reverse was observed for stomatal pore surface. Stomatal resistance increased in the urban and industrial land use class in comparison with the (sub-) urban green, but differences between land use classes were less pronounced. Spatial distribution maps for these leaf characteristics showed a high spatial variation, related to differences in habitat quality within the city. Hence, stomatal density and stomatal pore surface are assumed to be potentially good bio-indicators for urban habitat quality. - Stomatal characteristics of Plantago lanceolata can be used for biomonitoring of urban habitat quality.

Sophoraflavenone G Restricts Dengue and Zika Virus Infection via RNA Polymerase Interference.

Science.gov (United States)

Sze, Alexandre; Olagnier, David; Hadj, Samar Bel; Han, Xiaoying; Tian, Xiao Hong; Xu, Hong-Tao; Yang, Long; Shi, Qingwen; Wang, Penghua; Wainberg, Mark A; Wu, Jian Hui; Lin, Rongtuan

2017-10-03

Flaviviruses including Zika, Dengue and Hepatitis C virus cause debilitating diseases in humans, and the former are emerging as global health concerns with no antiviral treatments. We investigated Sophora Flavecens , used in Chinese medicine, as a source for antiviral compounds. We isolated Sophoraflavenone G and found that it inhibited Hepatitis C replication, but not Sendai or Vesicular Stomatitis Virus. Pre- and post-infection treatments demonstrated anti-flaviviral activity against Dengue and Zika virus, via viral RNA polymerase inhibition. These data suggest that Sophoraflavenone G represents a promising candidate regarding anti-Flaviviridae research.

Preventive effects of Ancer 20 injection against radiation stomatitis

Energy Technology Data Exchange (ETDEWEB)

Watanabe, Naohiko; Nomura, Yasuya; Takano, Shinya (Showa Univ., Tokyo (Japan). School of Medicine) (and others)

1993-10-01

Ancer 20 was injected subcutaneously twice a day into 23 patients during the couse of radiation therapy for head and neck cancer, with the aim of preventing radiation stomatitis. Oral mucosa was assessed both subjectively and objectively, in addition to white blood cell counts. Objective findings of oral mucosa revealed grade I in 71%, grade II in 52%, grade III in 14%, and grade IV in 5%. The dose of irradiation needed to produce grade I in 50% was 22.8 Gy. Subjective findings revealed grade I in 67%, grade II in 33%, and grade III in 10%. Irradiation dose needed to produce grade I in 50% was 23.9 Gy. Mucosous damage was slight when the white blood cell count of 6,000/mm[sup 3] was maintained. According to the rate of leukopenia, this drug was effective in 86.4%. These findings showed that Ancer 20 injection is useful in maintaining white blood cell counts and in preventing radiation stomatitis associated with radiation therapy especially to the field of mucous membrane. There was inverse correlation between white blood cell counts and both the occurrence rate and degree of radiation stomatitis. It seemed necessary to maintain white blood cell counts to prevent radiation stomatitis. (N.K.).

Pharmacological factors in the saliva of blood-feeding insects. Implications for vesicular stomatitis epidemiology.

Science.gov (United States)

Tabachnick, W J

2000-01-01

Vesicular stomatitis (VS) epizootics in the Western United States have caused substantial economic losses to U.S. livestock industries in 1995, 1997, and 1998. The role of arthropods in transmitting VS to U.S. livestock is unclear. In particular, the impact of arthropod salivary gland factors in VS infections in livestock needs study. Pharmacological effects of arthropod salivary gland factors on animals are reviewed. The potential effects of arthropod saliva on the transmission and spread of VS virus to livestock in the Western U.S. is presented with emphasis on the biting midge, Culicoides sonorensis. Information is discussed with attention to vector potential of C. sonorensis, and its use as a model for evaluating insect salivary gland pharmacology on livestock response to VS.

First case report of vesicular stomatitis in the State of Paraíba, Brazil

Directory of Open Access Journals (Sweden)

Inácio José Clementino

2014-10-01

Full Text Available The present report describes the first case of vesicular stomatitis in the State of Paraíba, Brazil. Records from the Official Veterinary Services of the State of Paraíba were analyzed while responding to a suspected case of vesicular disease at a property (property I in the municipality of Pombal in which the cattle showed clinical signs and lesions of vesicular disease. Surveillance in the surrounding area revealed similar lesions in cattle at two other properties (II and III. Based on these events, the suspicion was considered well founded, and samples were collected for evaluation at the National Agricultural Laboratory of the State of Pará. The property was interdicted, and those located within a 3 km radius (perifocal from the focus were inspected. At property I, 42.86% (6/14 of the cattle showed vesicular disease lesions characterized by intense sialorrhea, ruptured oral vesicles, epithelial detachment of the tongue and muzzle, and vesicular lesions in the udder and interdigital space. Similar lesions were detected in cattle at properties II and III, affecting 80.95% (34/42 and 11.54% (3/26 of the animals, respectively. Tissue samples collected from the three properties were positive for the vesicular stomatitis virus (Indiana 3 subtype. The properties were monitored for 21 days after the last infected animal was cured, and afterwards, the three properties were released.

Solubilization of glycoproteins of envelope viruses by detergents

International Nuclear Information System (INIS)

Berezin, V.E.; Zaides, V.M.; Artamsnov, A.F.; Isaeva, E.S.; Zhdanov, V.M.

1986-01-01

The action of a number of known ionic and nonionic detergents, as well as the new nonionic detergent MESK, on envelope viruses was investigated. It was shown that the nonionic detergents MESK, Triton X-100, and octyl-β-D-glucopyranoside selectively solubilize the outer glycoproteins of the virus particles. The nonionic detergent MESK has the mildest action. Using MESK, purified glycoproteins of influenza, parainfluenza, Venezuelan equine encephalomyelitis, vesicular stomatitis, rabies, and herpes viruses were obtained. The procedure for obtaining glycoproteins includes incubation of the virus suspension with the detergent MESK, removal of subvirus structures by centrifuging, and purification of glycoproteins from detergents by dialysis. Isolated glycoproteins retain a native structure and biological activity and possess high immunogenicity. The detergent MESK is promising for laboratory tests and with respect to the production of subunit vaccines

Reviewing host proteins of Rhabdoviridae: possible leads for lesser studied viruses.

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Guleria, A; Kiranmayi, M; Sreejith, R; Kumar, K; Sharma, S K; Gupta, S

2011-12-01

Rhabdoviridae, characterized by bullet-shaped viruses, is known for its diverse host range, which includes plants, arthropods, fishes and humans. Understanding the viral-host interactions of this family can prove beneficial in developing effective therapeutic strategies. The host proteins interacting with animal rhabdoviruses have been reviewed in this report. Several important host proteins commonly interacting with animal rhabdoviruses are being reported, some of which, interestingly, have molecular features, which can serve as potential antiviral targets. This review not only provides the generalized importance of the functions of animal rhabdovirus-associated host proteins for the first time but also compares them among the two most studied viruses, i.e. Rabies virus (RV) and Vesicular Stomatitis virus (VSV). The comparative data can be used for studying emerging viruses such as Chandipura virus (CHPV) and the lesser studied viruses such as Piry virus (PIRYV) and Isfahan virus (ISFV) of the Rhabdoviridae family.

Perforin and IFN-gamma do not significantly regulate the virus-specific CD8+ T cell response in the absence of antiviral effector activity

DEFF Research Database (Denmark)

Christensen, Jeanette Erbo; Wodarz, Dominik; Christensen, Jan P

2004-01-01

Using gene-targeted mice we have investigated whether perforin and/or interferon-gamma exert a direct regulatory effect on the expansion and contraction of antigen-specific CD8(+) T cells following infection with a virus (vesicular stomatitis virus) which is not controlled through these molecular...

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.

Curcumin modified silver nanoparticles for highly efficient inhibition of respiratory syncytial virus infection

Science.gov (United States)

Yang, Xiao Xi; Li, Chun Mei; Huang, Cheng Zhi

2016-01-01

Interactions between nanoparticles and viruses have attracted increasing attention due to the antiviral activity of nanoparticles and the resulting possibility to be employed as biomedical interventions. In this contribution, we developed a very simple route to prepare uniform and stable silver nanoparticles (AgNPs) with antiviral properties by using curcumin, which is a member of the ginger family isolated from rhizomes of the perennial herb Curcuma longa and has a wide range of biological activities like antioxidant, antifungal, antibacterial and anti-inflammatory effects, and acts as reducing and capping agents in this synthetic route. The tissue culture infectious dose (TCID50) assay showed that the curcumin modified silver nanoparticles (cAgNPs) have a highly efficient inhibition effect against respiratory syncytial virus (RSV) infection, giving a decrease of viral titers about two orders of magnitude at the concentration of cAgNPs under which no toxicity was found to the host cells. Mechanism investigations showed that cAgNPs could prevent RSV from infecting the host cells by inactivating the virus directly, indicating that cAgNPs are a novel promising efficient virucide for RSV.Interactions between nanoparticles and viruses have attracted increasing attention due to the antiviral activity of nanoparticles and the resulting possibility to be employed as biomedical interventions. In this contribution, we developed a very simple route to prepare uniform and stable silver nanoparticles (AgNPs) with antiviral properties by using curcumin, which is a member of the ginger family isolated from rhizomes of the perennial herb Curcuma longa and has a wide range of biological activities like antioxidant, antifungal, antibacterial and anti-inflammatory effects, and acts as reducing and capping agents in this synthetic route. The tissue culture infectious dose (TCID50) assay showed that the curcumin modified silver nanoparticles (cAgNPs) have a highly efficient inhibition

Stomatal Function Requires Pectin De-methyl-esterification of the Guard Cell Wall.

Science.gov (United States)

Amsbury, Sam; Hunt, Lee; Elhaddad, Nagat; Baillie, Alice; Lundgren, Marjorie; Verhertbruggen, Yves; Scheller, Henrik V; Knox, J Paul; Fleming, Andrew J; Gray, Julie E

2016-11-07

Stomatal opening and closure depends on changes in turgor pressure acting within guard cells to alter cell shape [1]. The extent of these shape changes is limited by the mechanical properties of the cells, which will be largely dependent on the structure of the cell walls. Although it has long been observed that guard cells are anisotropic due to differential thickening and the orientation of cellulose microfibrils [2], our understanding of the composition of the cell wall that allows them to undergo repeated swelling and deflation remains surprisingly poor. Here, we show that the walls of guard cells are rich in un-esterified pectins. We identify a pectin methylesterase gene, PME6, which is highly expressed in guard cells and required for stomatal function. pme6-1 mutant guard cells have walls enriched in methyl-esterified pectin and show a decreased dynamic range in response to triggers of stomatal opening/closure, including elevated osmoticum, suggesting that abrogation of stomatal function reflects a mechanical change in the guard cell wall. Altered stomatal function leads to increased conductance and evaporative cooling, as well as decreased plant growth. The growth defect of the pme6-1 mutant is rescued by maintaining the plants in elevated CO 2 , substantiating gas exchange analyses, indicating that the mutant stomata can bestow an improved assimilation rate. Restoration of PME6 rescues guard cell wall pectin methyl-esterification status, stomatal function, and plant growth. Our results establish a link between gene expression in guard cells and their cell wall properties, with a corresponding effect on stomatal function and plant physiology. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

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

Organ-specific effects of brassinosteroids on stomatal production coordinate with the action of Too Many Mouths.

Science.gov (United States)

Wang, Ming; Yang, Kezhen; Le, Jie

2015-03-01

In Arabidopsis, stomatal development initiates after protodermal cells acquire stomatal lineage cell fate. Stomata or their precursors communicate with their neighbor epidermal cells to ensure the "one cell spacing" rule. The signals from EPF/EPFL peptide ligands received by Too Many Mouths (TMM) and ERECTA-family receptors are supposed to be transduced by YODA MAPK cascade. A basic helix-loop-helix transcription factor SPEECHLESS (SPCH) is another key regulator of stomatal cell fate determination and asymmetric entry divisions, and SPCH activity is regulated by YODA MAPK cascade. Brassinosteroid (BR) signaling, one of the most well characterized signal transduction pathways in plants, contributes to the control of stomatal production. But opposite organ-specific effects of BR on stomatal production were reported. Here we confirm that stomatal production in hypocotyls is controlled by BR levels. YODA and CYCD4 are not essential for BR stomata-promoting function. Furthermore, we found that BR could confer tmm hypocotyls clustered stomatal phenotype, indicating that the BR organ-specific effects on stomatal production might coordinate with the TMM organ-specific actions. © 2014 Institute of Botany, Chinese Academy of Sciences.

Simulation of Stomatal Conductance and Water Use Efficiency of Tomato Leaves Exposed to Different Irrigation Regimes and Air CO2 Concentrations by a Modified "Ball-Berry" Model.

Science.gov (United States)

Wei, Zhenhua; Du, Taisheng; Li, Xiangnan; Fang, Liang; Liu, Fulai

2018-01-01

Stomatal conductance ( g s ) and water use efficiency ( WUE ) of tomato leaves exposed to different irrigation regimes and at ambient CO 2 ( a [CO 2 ], 400 ppm) and elevated CO 2 ( e [CO 2 ], 800 ppm) environments were simulated using the "Ball-Berry" model (BB-model). Data obtained from a preliminary experiment (Exp. I) was used for model parameterization, where measurements of leaf gas exchange of potted tomatoes were done during progressive soil drying for 5 days. The measured photosynthetic rate ( P n ) was used as an input for the model. Considering the effect of soil water deficits on g s , an equation modifying the slope ( m ) based on the mean soil water potential (Ψ s ) in the whole root zone was introduced. Compared to the original BB-model, the modified model showed greater predictability for both g s and WUE of tomato leaves at each [CO 2 ] growth environment. The models were further validated with data obtained from an independent experiment (Exp. II) where plants were subjected to three irrigation regimes: full irrigation (FI), deficit irrigation (DI), and alternative partial root-zone irrigation (PRI) for 40 days at both a [CO 2 ] and e [CO 2 ] environment. The simulation results indicated that g s was independently acclimated to e [CO 2 ] from P n . The modified BB-model performed better in estimating g s and WUE , especially for PRI strategy at both [CO 2 ] environments. A greater WUE could be seen in plants grown under e [CO 2 ] associated with PRI regime. Conclusively, the modified BB-model was capable of predicting g s and WUE of tomato leaves in various irrigation regimes at both a [CO 2 ] and e [CO 2 ] environments. This study could provide valuable information for better predicting plant WUE adapted to the future water-limited and CO 2 enriched environment.

An unexpected antibody response to an engineered influenza virus modifies CD8+ T cell responses.

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Thomas, Paul G; Brown, Scott A; Yue, Wen; So, Jenny; Webby, Richard J; Doherty, Peter C

2006-02-21

The ovalbumin(323-339) peptide that binds H2I-A(b) was engineered into the globular heads of hemagglutinin (H) molecules from serologically non-cross-reactive H1N1 and H3N2 influenza A viruses, the aim being to analyze recall CD4+ T cell responses in a virus-induced respiratory disease. Prime/challenge experiments with these H1ova and H3ova viruses in H2(b) mice gave the predicted, ovalbumin-specific CD4+ T cell response but showed an unexpectedly enhanced, early expansion of viral epitope-specific CD8+ T cells in spleen and a greatly diminished inflammatory process in the virus-infected respiratory tract. At the same time, the primary antibody response to the H3N2 challenge virus was significantly reduced, an effect that has been associated with preexisting neutralizing antibody in other experimental systems. Analysis of serum from the H1ova-primed mice showed low-level binding to H3ova but not to the wild-type H3N2 virus. Experiments with CD4+ T cell-depleted and Ig-/- mice indicated that this cross-reactive Ig is indeed responsible for the modified pathogenesis after respiratory challenge. Furthermore, the effect does not seem to be virus-dose related, although it does require infection. These findings suggest intriguing possibilities for vaccination and, at the same time, emphasize that engineered modifications in viruses may have unintended immunological consequences.

Fern Stomatal Responses to ABA and CO2 Depend on Species and Growth Conditions.

Science.gov (United States)

Hõrak, Hanna; Kollist, Hannes; Merilo, Ebe

2017-06-01

Changing atmospheric CO 2 levels, climate, and air humidity affect plant gas exchange that is controlled by stomata, small pores on plant leaves and stems formed by guard cells. Evolution has shaped the morphology and regulatory mechanisms governing stomatal movements to correspond to the needs of various land plant groups over the past 400 million years. Stomata close in response to the plant hormone abscisic acid (ABA), elevated CO 2 concentration, and reduced air humidity. Whether the active regulatory mechanisms that control stomatal closure in response to these stimuli are present already in mosses, the oldest plant group with stomata, or were acquired more recently in angiosperms remains controversial. It has been suggested that the stomata of the basal vascular plants, such as ferns and lycophytes, close solely hydropassively. On the other hand, active stomatal closure in response to ABA and CO 2 was found in several moss, lycophyte, and fern species. Here, we show that the stomata of two temperate fern species respond to ABA and CO 2 and that an active mechanism of stomatal regulation in response to reduced air humidity is present in some ferns. Importantly, fern stomatal responses depend on growth conditions. The data indicate that the stomatal behavior of ferns is more complex than anticipated before, and active stomatal regulation is present in some ferns and has possibly been lost in others. Further analysis that takes into account fern species, life history, evolutionary age, and growth conditions is required to gain insight into the evolution of land plant stomatal responses. © 2017 American Society of Plant Biologists. All Rights Reserved.

Expression of Arabidopsis Hexokinase in Citrus Guard Cells Controls Stomatal Aperture and Reduces Transpiration.

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Lugassi, Nitsan; Kelly, Gilor; Fidel, Lena; Yaniv, Yossi; Attia, Ziv; Levi, Asher; Alchanatis, Victor; Moshelion, Menachem; Raveh, Eran; Carmi, Nir; Granot, David

2015-01-01

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.

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.

Stomatal vs. genome size in angiosperms: the somatic tail wagging the genomic dog?

Science.gov (United States)

Hodgson, J G; Sharafi, M; Jalili, A; Díaz, S; Montserrat-Martí, G; Palmer, C; Cerabolini, B; Pierce, S; Hamzehee, B; Asri, Y; Jamzad, Z; Wilson, P; Raven, J A; Band, S R; Basconcelo, S; Bogard, A; Carter, G; Charles, M; Castro-Díez, P; Cornelissen, J H C; Funes, G; Jones, G; Khoshnevis, M; Pérez-Harguindeguy, N; Pérez-Rontomé, M C; Shirvany, F A; Vendramini, F; Yazdani, S; Abbas-Azimi, R; Boustani, S; Dehghan, M; Guerrero-Campo, J; Hynd, A; Kowsary, E; Kazemi-Saeed, F; Siavash, B; Villar-Salvador, P; Craigie, R; Naqinezhad, A; Romo-Díez, A; de Torres Espuny, L; Simmons, E

2010-04-01

Genome size is a function, and the product, of cell volume. As such it is contingent on ecological circumstance. The nature of 'this ecological circumstance' is, however, hotly debated. Here, we investigate for angiosperms whether stomatal size may be this 'missing link': the primary determinant of genome size. Stomata are crucial for photosynthesis and their size affects functional efficiency. Stomatal and leaf characteristics were measured for 1442 species from Argentina, Iran, Spain and the UK and, using PCA, some emergent ecological and taxonomic patterns identified. Subsequently, an assessment of the relationship between genome-size values obtained from the Plant DNA C-values database and measurements of stomatal size was carried out. Stomatal size is an ecologically important attribute. It varies with life-history (woody species < herbaceous species < vernal geophytes) and contributes to ecologically and physiologically important axes of leaf specialization. Moreover, it is positively correlated with genome size across a wide range of major taxa. Stomatal size predicts genome size within angiosperms. Correlation is not, however, proof of causality and here our interpretation is hampered by unexpected deficiencies in the scientific literature. Firstly, there are discrepancies between our own observations and established ideas about the ecological significance of stomatal size; very large stomata, theoretically facilitating photosynthesis in deep shade, were, in this study (and in other studies), primarily associated with vernal geophytes of unshaded habitats. Secondly, the lower size limit at which stomata can function efficiently, and the ecological circumstances under which these minute stomata might occur, have not been satisfactorally resolved. Thus, our hypothesis, that the optimization of stomatal size for functional efficiency is a major ecological determinant of genome size, remains unproven.

A comparison of two stomatal conductance models for ozone flux modelling using data from two Brassica species

International Nuclear Information System (INIS)

Op de Beeck, M.; De Bock, M.; Vandermeiren, K.; Temmerman, L. de; Ceulemans, R.

2010-01-01

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 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 st variance, respectively. For broccoli, the models were able to explain 53% and 51% of the observed g st variance, respectively. These results support the coupled semi-empirical stomatal-photosynthesis model as a valid alternative to the multiplicative stomatal model for O 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.

Vesicular stomatitis forecasting based on Google Trends.

Science.gov (United States)

Wang, JianYing; Zhang, Tong; Lu, Yi; Zhou, GuangYa; Chen, Qin; Niu, Bing

2018-01-01

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. 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. 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. 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.

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

Vesicular stomatitis forecasting based on Google Trends.

Directory of Open Access Journals (Sweden)

JianYing Wang

Full Text Available 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.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.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.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.

Use of modified live feline panleukopenia virus vaccine to immunize dogs against canine parvovirus.

Science.gov (United States)

Pollock, R V; Carmichael, L E

1983-02-01

Modified live feline panleukopenia virus (FPLV) vaccine protected dogs against canine parvovirus (CPV) infection. However, unlike the long-lived (greater than or equal to 20-month) immunity engendered by CPV infection, the response of dogs to living FPLV was variable. Doses of FPLV (snow leopard strain) in excess of 10(5.7) TCID50 were necessary for uniform immunization; smaller inocula resulted in decreased success rates. The duration of immunity, as measured by the persistence of hemagglutination-inhibiting antibody, was related to the magnitude of the initial response to vaccination; dogs with vigorous initial responses resisted oronasal CPV challenge exposure 6 months after vaccination, and hemagglutination-inhibiting antibodies persisted in such dogs for greater than 1 year. Limited replication of FPLV in dogs was demonstrated, but unlike CPV, the feline virus did not spread to contact dogs or cats. Adverse reactions were not associated with living FPLV vaccination, and FPLV did not interfere with simultaneous response to attenuated canine distemper virus.

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.

Modified vaccinia virus Ankara expressing the hemagglutinin of pandemic (H1N1) 2009 virus induces cross-protective immunity against Eurasian 'avian-like' H1N1 swine viruses in mice.

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Castrucci, Maria R; Facchini, Marzia; Di Mario, Giuseppina; Garulli, Bruno; Sciaraffia, Ester; Meola, Monica; Fabiani, Concetta; De Marco, Maria A; Cordioli, Paolo; Siccardi, Antonio; Kawaoka, Yoshihiro; Donatelli, Isabella

2014-05-01

To examine cross-reactivity between hemagglutinin (HA) derived from A/California/7/09 (CA/09) virus and that derived from representative Eurasian "avian-like" (EA) H1N1 swine viruses isolated in Italy between 1999 and 2008 during virological surveillance in pigs. Modified vaccinia virus Ankara (MVA) expressing the HA gene of CA/09 virus (MVA-HA-CA/09) was used as a vaccine to investigate cross-protective immunity against H1N1 swine viruses in mice. Two classical swine H1N1 (CS) viruses and four representative EA-like H1N1 swine viruses previously isolated during outbreaks of respiratory disease in pigs on farms in Northern Italy were used in this study. Female C57BL/6 mice were vaccinated with MVA/HA/CA/09 and then challenged intranasally with H1N1 swine viruses. Cross-reactive antibody responses were determined by hemagglutination- inhibition (HI) and virus microneutralizing (MN) assays of sera from MVA-vaccinated mice. The extent of protective immunity against infection with H1N1 swine viruses was determined by measuring lung viral load on days 2 and 4 post-challenge. Systemic immunization of mice with CA/09-derived HA, vectored by MVA, elicited cross-protective immunity against recent EA-like swine viruses. This immune protection was related to the levels of cross-reactive HI antibodies in the sera of the immunized mice and was dependent on the similarity of the antigenic site Sa of H1 HAs. Our findings suggest that the herd immunity elicited in humans by the pandemic (H1N1) 2009 virus could limit the transmission of recent EA-like swine HA genes into the influenza A virus gene pool in humans. © 2013 The Authors Influenza and Other Respiratory Viruses Published by John Wiley & Sons Ltd.

A steady-state stomatal model of balanced leaf gas exchange, hydraulics and maximal source-sink flux.

Science.gov (United States)

Hölttä, Teemu; Lintunen, Anna; Chan, Tommy; Mäkelä, Annikki; Nikinmaa, Eero

2017-07-01

Trees must simultaneously balance their CO2 uptake rate via stomata, photosynthesis, the transport rate of sugars and rate of sugar utilization in sinks while maintaining a favourable water and carbon balance. We demonstrate using a numerical model that it is possible to understand stomatal functioning from the viewpoint of maximizing the simultaneous photosynthetic production, phloem transport and sink sugar utilization rate under the limitation that the transpiration-driven hydrostatic pressure gradient sets for those processes. A key feature in our model is that non-stomatal limitations to photosynthesis increase with decreasing leaf water potential and/or increasing leaf sugar concentration and are thus coupled to stomatal conductance. Maximizing the photosynthetic production rate using a numerical steady-state model leads to stomatal behaviour that is able to reproduce the well-known trends of stomatal behaviour in response to, e.g., light, vapour concentration difference, ambient CO2 concentration, soil water status, sink strength and xylem and phloem hydraulic conductance. We show that our results for stomatal behaviour are very similar to the solutions given by the earlier models of stomatal conductance derived solely from gas exchange considerations. Our modelling results also demonstrate how the 'marginal cost of water' in the unified stomatal conductance model and the optimal stomatal model could be related to plant structural and physiological traits, most importantly, the soil-to-leaf hydraulic conductance and soil moisture. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

1995 epizootic of vesicular stomatitis (New Jersey serotype) in the western United States: an entomologic perspective.

Science.gov (United States)

Schmidtmann, E T; Tabachnick, W J; Hunt, G J; Thompson, L H; Hurd, H S

1999-01-01

Entomologic and epizootic data are reviewed concerning the potential for transmission of vesicular stomatitis (VS) virus by insects, including field data from case-positive premises in New Mexico and Colorado during the 1995 outbreak of the New Jersey serotype (VSNJ). As with previous outbreaks of VSNJ in the western United States, the 1995 epizootic illustrated that risk of exposure is seasonal, increasing during warm weather and decreasing with onset of cool weather; virus activity spread from south to north along river valleys of the southwestern and Rocky Mountain states; clinical disease was detected most commonly in horses, but also occurred in cattle and 1 llama; and most infections were subclinical. Overall, 367 case-positive premises were identified during the 1995 outbreak, with foci of virus activity along the Rio Grande River south of Albuquerque, NM, in southwestern Colorado, and along the Colorado River near Grand Junction, CO. The establishment of a 16-km (10-mile) radius zone of restricted animal movement around confirmed positive premises, along with imposition of state and international embargoes, created economic hardship for livestock owners and producers. The importance of defining the role of blood-feeding insects as biological vectors of VSNJ virus relative to risk factors that promote high levels of insect transmission, such as the presence of livestock along western river valleys, blood feeding activity, and frequent transport of animals for recreational purposes, is emphasized as a basis for developing effective disease management.

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.

Role of CD28 co-stimulation in generation and maintenance of virus-specific T cells

DEFF Research Database (Denmark)

Christensen, Jeanette Erbo; Christensen, Jan P; Kristensen, Nanna N

2002-01-01

Efficient induction of T cell responses is normally assumed to require both TCR-mediated signaling and engagement of co-stimulatory molecules, in particular CD28. However, the importance of CD28 co-stimulation in induction and maintenance of antiviral T cell responses is not clearly established....... For this reason antiviral CD4(+) and CD8(+) T cell responses in CD28-deficient mice were studied using two different viruses [vesicular stomatitis virus and lymphocytic choriomeningitis virus (LCMV)]. Intracellular cytokine staining and/or MHC-peptide tetramers were used to enumerate antigen-specific T cells...

Modified Vaccinia Virus Ankara: History, Value in Basic Research, and Current Perspectives for Vaccine Development.

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Volz, A; Sutter, G

2017-01-01

Safety tested Modified Vaccinia virus Ankara (MVA) is licensed as third-generation vaccine against smallpox and serves as a potent vector system for development of new candidate vaccines against infectious diseases and cancer. Historically, MVA was developed by serial tissue culture passage in primary chicken cells of vaccinia virus strain Ankara, and clinically used to avoid the undesirable side effects of conventional smallpox vaccination. Adapted to growth in avian cells MVA lost the ability to replicate in mammalian hosts and lacks many of the genes orthopoxviruses use to conquer their host (cell) environment. As a biologically well-characterized mutant virus, MVA facilitates fundamental research to elucidate the functions of poxvirus host-interaction factors. As extremely safe viral vectors MVA vaccines have been found immunogenic and protective in various preclinical infection models. Multiple recombinant MVA currently undergo clinical testing for vaccination against human immunodeficiency viruses, Mycobacterium tuberculosis or Plasmodium falciparum. The versatility of the MVA vector vaccine platform is readily demonstrated by the swift development of experimental vaccines for immunization against emerging infections such as the Middle East Respiratory Syndrome. Recent advances include promising results from the clinical testing of recombinant MVA-producing antigens of highly pathogenic avian influenza virus H5N1 or Ebola virus. This review summarizes our current knowledge about MVA as a unique strain of vaccinia virus, and discusses the prospects of exploiting this virus as research tool in poxvirus biology or as safe viral vector vaccine to challenge existing and future bottlenecks in vaccinology. © 2017 Elsevier Inc. All rights reserved.

A fusion-inhibiting peptide against Rift Valley fever virus inhibits multiple, diverse viruses.

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Jeffrey W Koehler

Full Text Available For enveloped viruses, fusion of the viral envelope with a cellular membrane is critical for a productive infection to occur. This fusion process is mediated by at least three classes of fusion proteins (Class I, II, and III based on the protein sequence and structure. For Rift Valley fever virus (RVFV, the glycoprotein Gc (Class II fusion protein mediates this fusion event following entry into the endocytic pathway, allowing the viral genome access to the cell cytoplasm. Here, we show that peptides analogous to the RVFV Gc stem region inhibited RVFV infectivity in cell culture by inhibiting the fusion process. Further, we show that infectivity can be inhibited for diverse, unrelated RNA viruses that have Class I (Ebola virus, Class II (Andes virus, or Class III (vesicular stomatitis virus fusion proteins using this single peptide. Our findings are consistent with an inhibition mechanism similar to that proposed for stem peptide fusion inhibitors of dengue virus in which the RVFV inhibitory peptide first binds to both the virion and cell membranes, allowing it to traffic with the virus into the endocytic pathway. Upon acidification and rearrangement of Gc, the peptide is then able to specifically bind to Gc and prevent fusion of the viral and endocytic membranes, thus inhibiting viral infection. These results could provide novel insights into conserved features among the three classes of viral fusion proteins and offer direction for the future development of broadly active fusion inhibitors.

Simulation of Stomatal Conductance and Water Use Efficiency of Tomato Leaves Exposed to Different Irrigation Regimes and Air CO2 Concentrations by a Modified “Ball-Berry” Model

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Zhenhua Wei

2018-04-01

Full Text Available Stomatal conductance (gs and water use efficiency (WUE of tomato leaves exposed to different irrigation regimes and at ambient CO2 (a[CO2], 400 ppm and elevated CO2 (e[CO2], 800 ppm environments were simulated using the “Ball-Berry” model (BB-model. Data obtained from a preliminary experiment (Exp. I was used for model parameterization, where measurements of leaf gas exchange of potted tomatoes were done during progressive soil drying for 5 days. The measured photosynthetic rate (Pn was used as an input for the model. Considering the effect of soil water deficits on gs, an equation modifying the slope (m based on the mean soil water potential (Ψs in the whole root zone was introduced. Compared to the original BB-model, the modified model showed greater predictability for both gs and WUE of tomato leaves at each [CO2] growth environment. The models were further validated with data obtained from an independent experiment (Exp. II where plants were subjected to three irrigation regimes: full irrigation (FI, deficit irrigation (DI, and alternative partial root-zone irrigation (PRI for 40 days at both a[CO2] and e[CO2] environment. The simulation results indicated that gs was independently acclimated to e[CO2] from Pn. The modified BB-model performed better in estimating gs and WUE, especially for PRI strategy at both [CO2] environments. A greater WUE could be seen in plants grown under e[CO2] associated with PRI regime. Conclusively, the modified BB-model was capable of predicting gs and WUE of tomato leaves in various irrigation regimes at both a[CO2] and e[CO2] environments. This study could provide valuable information for better predicting plant WUE adapted to the future water-limited and CO2 enriched environment.

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.

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 ...

Enhanced light microscopy visualization of virus particles from Zika virus to filamentous ebolaviruses.

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George G Daaboul

Full Text Available Light microscopy is a powerful tool in the detection and analysis of parasites, fungi, and prokaryotes, but has been challenging to use for the detection of individual virus particles. Unlabeled virus particles are too small to be visualized using standard visible light microscopy. Characterization of virus particles is typically performed using higher resolution approaches such as electron microscopy or atomic force microscopy. These approaches require purification of virions away from their normal millieu, requiring significant levels of expertise, and can only enumerate small numbers of particles per field of view. Here, we utilize a visible light imaging approach called Single Particle Interferometric Reflectance Imaging Sensor (SP-IRIS that allows automated counting and sizing of thousands of individual virions. Virions are captured directly from complex solutions onto a silicon chip and then detected using a reflectance interference imaging modality. We show that the use of different imaging wavelengths allows the visualization of a multitude of virus particles. Using Violet/UV illumination, the SP-IRIS technique is able to detect individual flavivirus particles (~40 nm, while green light illumination is capable of identifying and discriminating between vesicular stomatitis virus and vaccinia virus (~360 nm. Strikingly, the technology allows the clear identification of filamentous infectious ebolavirus particles and virus-like particles. The ability to differentiate and quantify unlabeled virus particles extends the usefulness of traditional light microscopy and can be embodied in a straightforward benchtop approach allowing widespread applications ranging from rapid detection in biological fluids to analysis of virus-like particles for vaccine development and production.

Functional Analysis of Cellulose and Xyloglucan in the Walls of Stomatal Guard Cells of Arabidopsis1[OPEN

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Rui, Yue; Anderson, Charles T.

2016-01-01

Stomatal guard cells are pairs of specialized epidermal cells that control water and CO2 exchange between the plant and the environment. To fulfill the functions of stomatal opening and closure that are driven by changes in turgor pressure, guard cell walls must be both strong and flexible, but how the structure and dynamics of guard cell walls enable stomatal function remains poorly understood. To address this question, we applied cell biological and genetic analyses to investigate guard cell walls and their relationship to stomatal function in Arabidopsis (Arabidopsis thaliana). Using live-cell spinning disk confocal microscopy, we measured the motility of cellulose synthase (CESA)-containing complexes labeled by green fluorescent protein (GFP)-CESA3 and observed a reduced proportion of GFP-CESA3 particles colocalizing with microtubules upon stomatal closure. Imaging cellulose organization in guard cells revealed a relatively uniform distribution of cellulose in the open state and a more fibrillar pattern in the closed state, indicating that cellulose microfibrils undergo dynamic reorganization during stomatal movements. In cesa3je5 mutants defective in cellulose synthesis and xxt1 xxt2 mutants lacking the hemicellulose xyloglucan, stomatal apertures, changes in guard cell length, and cellulose reorganization were aberrant during fusicoccin-induced stomatal opening or abscisic acid-induced stomatal closure, indicating that sufficient cellulose and xyloglucan are required for normal guard cell dynamics. Together, these results provide new insights into how guard cell walls allow stomata to function as responsive mediators of gas exchange at the plant surface. PMID:26729799

Drought limitations to leaf-level gas exchange: results from a model linking stomatal optimization and cohesion-tension theory.

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Novick, Kimberly A; Miniat, Chelcy F; Vose, James M

2016-03-01

We merge concepts from stomatal optimization theory and cohesion-tension theory to examine the dynamics of three mechanisms that are potentially limiting to leaf-level gas exchange in trees during drought: (1) a 'demand limitation' driven by an assumption of optimal stomatal functioning; (2) 'hydraulic limitation' of water movement from the roots to the leaves; and (3) 'non-stomatal' limitations imposed by declining leaf water status within the leaf. Model results suggest that species-specific 'economics' of stomatal behaviour may play an important role in differentiating species along the continuum of isohydric to anisohydric behaviour; specifically, we show that non-stomatal and demand limitations may reduce stomatal conductance and increase leaf water potential, promoting wide safety margins characteristic of isohydric species. We used model results to develop a diagnostic framework to identify the most likely limiting mechanism to stomatal functioning during drought and showed that many of those features were commonly observed in field observations of tree water use dynamics. Direct comparisons of modelled and measured stomatal conductance further indicated that non-stomatal and demand limitations reproduced observed patterns of tree water use well for an isohydric species but that a hydraulic limitation likely applies in the case of an anisohydric species. Published 2015. This article is a US Government work and is in the public domain in the USA.

Contrasting responses of leaf stomatal characteristics to climate change: a considerable challenge to predict carbon and water cycles.

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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 CO 2 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 CO 2 concentration, temperature, and soil water availability. Moreover, threshold effects of elevated CO 2 and temperature on stomatal frequency were detected, indicating that the response of stomatal density to increasing CO 2 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 CO 2 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.

Reactive oxygen species signaling and stomatal movement: Current updates and future perspectives

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Rachana Singh

2017-04-01

Full Text Available Reactive oxygen species (ROS, a by-product of aerobic metabolism were initially studied in context to their damaging effect but recent decades witnessed significant advancements in understanding the role of ROS as signaling molecules. Contrary to earlier views, it is becoming evident that ROS production is not necessarily a symptom of cellular dysfunction but it might represent a necessary signal in adjusting the cellular machinery according to the altered conditions. Stomatal movement is controlled by multifaceted signaling network in response to endogenous and environmental signals. Furthermore, the stomatal aperture is regulated by a coordinated action of signaling proteins, ROS-generating enzymes, and downstream executors like transporters, ion pumps, plasma membrane channels, which control the turgor pressure of the guard cell. The earliest hallmarks of stomatal closure are ROS accumulation in the apoplast and chloroplasts and thereafter, there is a successive increase in cytoplasmic Ca2+ level which rules the multiple kinases activity that in turn regulates the activity of ROS-generating enzymes and various ion channels. In addition, ROS also regulate the action of multiple proteins directly by oxidative post translational modifications to adjust guard cell signaling. Notwithstanding, an active progress has been made with ROS signaling mechanism but the regulatory action for ROS signaling processes in stomatal movement is still fragmentary. Therefore, keeping in view the above facts, in this mini review the basic concepts and role of ROS signaling in the stomatal movement have been presented comprehensively along with recent highlights.

Ebola virus (EBOV) infection: Therapeutic strategies.

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De Clercq, Erik

2015-01-01

Within less than a year after its epidemic started (in December 2013) in Guinea, Ebola virus (EBOV), a member of the filoviridae, has spread over a number of West-African countries (Guinea, Sierra Leone and Liberia) and gained allures that have been unprecedented except by human immunodeficiency virus (HIV). Although EBOV is highly contagious and transmitted by direct contact with body fluids, it could be counteracted by the adequate chemoprophylactic and -therapeutic interventions: vaccines, antibodies, siRNAs (small interfering RNAs), interferons and chemical substances, i.e. neplanocin A derivatives (i.e. 3-deazaneplanocin A), BCX4430, favipiravir (T-705), endoplasmic reticulum (ER) α-glucosidase inhibitors and a variety of compounds that have been found to inhibit EBOV infection blocking viral entry or by a mode of action that still has to be resolved. Much has to be learned from the mechanism of action of the compounds active against VSV (vesicular stomatitis virus), a virus belonging to the rhabdoviridae, that in its mode of replication could be exemplary for the replication of filoviridae. Copyright © 2014 Elsevier Inc. All rights reserved.

Nitric oxide in guard cells as an important secondary messenger during stomatal closure

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Gunja eGayatri

2013-10-01

Full Text Available he modulation of guard cell function is the basis of stomatal closure, essential for optimizing water use and CO2 uptake by leaves. Nitric oxide (NO in guard cells plays a very important role as a secondary messenger during stomatal closure induced by effectors, including hormones. For example, exposure to abscisic acid (ABA triggers a marked increase in NO of guard cells, well before stomatal closure. In guard cells of multiple species, like Arabidopsis, Vicia and pea, exposure to ABA or methyl jasmonate or even microbial elicitors (e.g. chitosan induces production of NO as well as reactive oxygen species (ROS. The role of NO in stomatal closure has been confirmed by using NO donors (e.g. SNP and NO scavengers (like cPTIO and inhibitors of NOS (L-NAME or NR (tungstate. Two enzymes: a L-NAME-sensitive, nitric oxide synthase (NOS-like enzyme and a tungstate-sensitive nitrate reductase (NR, can mediate ABA-induced NO rise in guard cells. However, the existence of true NOS in plant tissues and its role in guard cell NO-production are still a matter of intense debate. Guard cell signal transduction leading to stomatal closure involves the participation of several components, besides NO, such as cytosolic pH, ROS, free Ca2+ and phospholipids. Use of fluorescent dyes has revealed that the rise in NO of guard cells occurs after the increase in cytoplasmic pH and ROS. The rise in NO causes an elevation in cytosolic free Ca2+ and promotes the efflux of cations as well as anions from guard cells. Stomatal guard cells have become a model system to study the signalling cascade mechanisms in plants, particularly with NO as a dominant component. The interrelationships and interactions of NO with cytosolic pH, ROS, and free Ca2+ are quite complex and need further detailed examination. While assessing critically the available literature, the present review projects possible areas of further work related to NO-action in stomatal guard cells.

Constitutive activation of a plasma membrane H+-ATPase prevents abscisic acid-mediated stomatal closure

Science.gov (United States)

Merlot, Sylvain; Leonhardt, Nathalie; Fenzi, Francesca; Valon, Christiane; Costa, Miguel; Piette, Laurie; Vavasseur, Alain; Genty, Bernard; Boivin, Karine; Müller, Axel; Giraudat, Jérôme; Leung, Jeffrey

2007-01-01

Light activates proton (H+)-ATPases in guard cells, to drive hyperpolarization of the plasma membrane to initiate stomatal opening, allowing diffusion of ambient CO2 to photosynthetic tissues. Light to darkness transition, high CO2 levels and the stress hormone abscisic acid (ABA) promote stomatal closing. The overall H+-ATPase activity is diminished by ABA treatments, but the significance of this phenomenon in relationship to stomatal closure is still debated. We report two dominant mutations in the OPEN STOMATA2 (OST2) locus of Arabidopsis that completely abolish stomatal response to ABA, but importantly, to a much lesser extent the responses to CO2 and darkness. The OST2 gene encodes the major plasma membrane H+-ATPase AHA1, and both mutations cause constitutive activity of this pump, leading to necrotic lesions. H+-ATPases have been traditionally assumed to be general endpoints of all signaling pathways affecting membrane polarization and transport. Our results provide evidence that AHA1 is a distinct component of an ABA-directed signaling pathway, and that dynamic downregulation of this pump during drought is an essential step in membrane depolarization to initiate stomatal closure. PMID:17557075

TCR Down-Regulation Controls Virus-Specific CD8+ T Cell Responses

DEFF Research Database (Denmark)

Bonefeld, Charlotte Menné; Haks, Mariëlle; Nielsen, Bodil

2008-01-01

The CD3gamma di-leucine-based motif plays a central role in TCR down-regulation. However, little is understood about the role of the CD3gamma di-leucine-based motif in physiological T cell responses. In this study, we show that the expansion in numbers of virus-specific CD8(+) T cells is impaired...... in mice with a mutated CD3gamma di-leucine-based motif. The CD3gamma mutation did not impair early TCR signaling, nor did it compromise recruitment or proliferation of virus-specific T cells, but it increased the apoptosis rate of the activated T cells by increasing down-regulation of the antiapoptotic...... molecule Bcl-2. This resulted in a 2-fold reduction in the clonal expansion of virus-specific CD8(+) T cells during the acute phase of vesicular stomatitis virus and lymphocytic choriomeningitis virus infections. These results identify an important role of CD3gamma-mediated TCR down-regulation in virus...

Effects of a Heat Wave on Nocturnal Stomatal Conductance in Eucalyptus camaldulensis

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Víctor Resco de Dios

2018-06-01

Full Text Available Nocturnal transpiration constitutes a significant yet poorly understood component of the global water cycle. Modeling nocturnal transpiration has been complicated by recent findings showing that stomata respond differently to environmental drivers over day- vs. night-time periods. Here, we propose that nocturnal stomatal conductance depends on antecedent daytime conditions. We tested this hypothesis across six genotypes of Eucalyptus camaldulensis Dehnh. growing under different CO2 concentrations (ambient vs. elevated and exposed to contrasting temperatures (ambient vs. heat wave for four days prior to the night of measurements, when all plants experienced ambient temperature conditions. We observed significant effects after the heat wave that led to 36% reductions in nocturnal stomatal conductance. The response was partly driven by changes in daytime stomatal behavior but additional factors may have come into play. We also observed significant differences in response to the heat wave across genotypes, likely driven by local adaptation to their climate of origin, but CO2 played no effect. Stomatal models may need to incorporate the role of antecedent effects to improve projections particularly after drastic changes in the environment such as heat waves.

Stomatal response of Pinus sylvestriformis to elevated CO2 concentrations during the four years of exposure

Institute of Scientific and Technical Information of China (English)

ZHOU Yu-mei; HAN Shi-jie; LIU Ying; JIA Xia

2005-01-01

Four-year-old Pinus sylvestriformis were exposed for four growing seasons in open top chambers to ambient CO2 concentration (approx. 350 μmol·mol-1) and high CO2 concentrations (500 and 700 μmol·mol-1) at Research Station of Changbai Mountain Forest Ecosystems, Chinese Academy of Sciences at Antu Town, Jilin Province, China (42oN, 128oE). Stomatal response to elevated CO2 concentrations was examined by stomatal conductance (gs), ratio of intercellular to ambient CO2 concentration (ci/ca) and stomatal number. Reciprocal transfer experiments of stomatal conductance showed that stomatal conductance in high-[CO2]-grown plants increased in comparison with ambient-[CO2]-grown plants when measured at their respective growth CO2 concentration and at the same measurement CO2 concentration (except a reduction in 700 μmol·mol-1 CO2 grown plants compared with plants on unchambered field when measured at growth CO2 concentration and 350 μmol·mol-1CO2). High-[CO2]-grown plants exhibited lower ci/ca ratios than ambient-[CO2]-grown plants when measured at their respective growth CO2 concentration. However, ci/ca ratios increased for plants grown in high CO2 concentrations compared with control plants when measured at the same CO2 concentration. There was no significant difference in stomatal number per unit long needle between elevated and ambient CO2. However, elevated CO2 concentrations reduced the total stomatal number of whole needle by the decline of stomatal line and changed the allocation pattern of stomata between upper and lower surface of needle.

Differential sensitivity of bat cells to infection by enveloped RNA viruses: coronaviruses, paramyxoviruses, filoviruses, and influenza viruses.

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Markus Hoffmann

Full Text Available Bats (Chiroptera host major human pathogenic viruses including corona-, paramyxo, rhabdo- and filoviruses. We analyzed six different cell lines from either Yinpterochiroptera (including African flying foxes and a rhinolophid bat or Yangochiroptera (genera Carollia and Tadarida for susceptibility to infection by different enveloped RNA viruses. None of the cells were sensitive to infection by transmissible gastroenteritis virus (TGEV, a porcine coronavirus, or to infection mediated by the Spike (S protein of SARS-coronavirus (SARS-CoV incorporated into pseudotypes based on vesicular stomatitis virus (VSV. The resistance to infection was overcome if cells were transfected to express the respective cellular receptor, porcine aminopeptidase N for TGEV or angiotensin-converting enzyme 2 for SARS-CoV. VSV pseudotypes containing the S proteins of two bat SARS-related CoV (Bg08 and Rp3 were unable to infect any of the six tested bat cell lines. By contrast, viral pseudotypes containing the surface protein GP of Marburg virus from the family Filoviridae infected all six cell lines though at different efficiency. Notably, all cells were sensitive to infection by two paramyxoviruses (Sendai virus and bovine respiratory syncytial virus and three influenza viruses from different subtypes. These results indicate that bat cells are more resistant to infection by coronaviruses than to infection by paramyxoviruses, filoviruses and influenza viruses. Furthermore, these results show a receptor-dependent restriction of the infection of bat cells by CoV. The implications for the isolation of coronaviruses from bats are discussed.

Curcumin inhibits Zika and chikungunya virus infection by inhibiting cell binding.

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Mounce, Bryan C; Cesaro, Teresa; Carrau, Lucia; Vallet, Thomas; Vignuzzi, Marco

2017-06-01

Several compounds extracted from spices and herbs exhibit antiviral effects in vitro, suggesting potential pharmacological uses. Curcumin, a component of turmeric, has been used as a food additive and herbal supplement due to its potential medicinal properties. Previously, curcumin exhibited antiviral properties against several viruses, including dengue virus and hepatitis C virus, among others. Here, we describe the antiviral effect of curcumin on Zika and chikungunya viruses, two mosquito-borne outbreak viruses. Both viruses responded to treatment of cells with up to 5 μM curumin without impacting cellular viability. We observed that direct treatment of virus with curcumin reduced infectivity of virus in a dose- and time-dependent manner for these enveloped viruses, as well as vesicular stomatitis virus. In contrast, we found no change in infectivity for Coxsackievirus B3, a non-enveloped virus. Derivatives of curcumin also exhibited antiviral activity against enveloped viruses. Further examination revealed that curcumin interfered with the binding of the enveloped viruses to cells in a dose-dependent manner, though the integrity of the viral RNA was maintained. Together, these results expand the family of viruses sensitive to curcumin and provide a mechanism of action for curcumin's effect on these enveloped viruses. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

Ozone slows stomatal response to light and leaf wounding in a Mediterranean evergreen broadleaf, Arbutus unedo.

Science.gov (United States)

Paoletti, Elena

2005-04-01

The effect of a 90-d ozone exposure (charcoal-filtered air or 110 nmol mol(-1) O3) on stomatal conductance (gs) was investigated in the Mediterranean evergreen broadleaf Arbutus unedo L. Ozone did not significantly reduce midday steady-state gs compared to controls. However, it slowed stomatal response to abrupt reduction of light intensity and to increasing water stress, applied by severing the leaf midrib. Ozone slowed stomatal closure, rather than aperture. Nevertheless, vein-cutting did not allow ozonated leaves to reach the pre-injury gs levels, like controls did, suggesting re-opening was still, slowly in progress. The sluggish behaviour was recorded 10 days after cessation of O3 exposure ("memory effect") and may affect stomatal control in response to sunflecks and leaf wounding. Mediterranean evergreen broadleaves are regarded as tolerant to O3 exposure. Nevertheless, measurements of steady-state gs at midday may not account for altered stomatal responses to stressors.

Ethylene limits abscisic acid- or soil drying-induced stomatal closure in aged wheat leaves.

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Chen, Lin; Dodd, Ian C; Davies, William J; Wilkinson, Sally

2013-10-01

The mechanism of age-induced decreased stomatal sensitivity to abscisic acid (ABA) and soil drying has been explored here. Older, fully expanded leaves partly lost their ability to close stomata in response to foliar ABA sprays, and soil drying which stimulated endogenous ABA production, while young fully expanded leaves closed their stomata more fully. However, ABA- or soil drying-induced stomatal closure of older leaves was partly restored by pretreating plants with 1-methylcyclopropene (1-MCP), which can antagonize ethylene receptors, or by inoculating soil around the roots with the rhizobacterium Variovorax paradoxus 5C-2, which contains 1-aminocyclopropane-1-carboxylic acid (ACC)-deaminase. ACC (the immediate biosynthetic precursor of ethylene) sprays revealed higher sensitivity of stomata to ethylene in older leaves than younger leaves, despite no differences in endogenous ACC concentrations or ethylene emission. Taken together, these results indicate that the relative insensitivity of stomatal closure to ABA and soil drying in older leaves is likely due to altered stomatal sensitivity to ethylene, rather than ethylene production. To our knowledge, this is the first study to mechanistically explain diminished stomatal responses to soil moisture deficit in older leaves, and the associated reduction in leaf water-use efficiency. © 2013 John Wiley & Sons Ltd.

The effect of competition from neighbours on stomatal conductance in lettuce and tomato plants.

Science.gov (United States)

Vysotskaya, Lidiya; Wilkinson, Sally; Davies, William J; Arkhipova, Tatyana; Kudoyarova, Guzel

2011-05-01

Competition decreased transpiration from young lettuce plants after 2 days, before any reductions in leaf area became apparent, and stomatal conductance (g(s) ) of lettuce and tomato plants was also reduced. Stomatal closure was not due to hydraulic signals or competition for nutrients, as soil water content, leaf water status and leaf nitrate concentrations were unaffected by neighbours. Competition-induced stomatal closure was absent in an abscisic acid (ABA)-deficient tomato mutant, flacca, indicating a fundamental involvement of ABA. Although tomato xylem sap ABA concentrations were unaffected by the presence of neighbours, ABA/pH-based stomatal modulation is still likely to underlie the response to competition, as soil and xylem sap alkalization was observed in competing plants. Competition also modulated leaf ethylene production, and treatment of lettuce plants with an ethylene perception inhibitor (1-methylcyclopropene) diminished the difference in g(s) between single and competing plants grown in a controlled environment room, but increased it in plants grown in the greenhouse: ethylene altered the extent of the stomatal response to competition. Effects of competition on g(s) are discussed in terms of the detection of the absence of neighbours: increases in g(s) and carbon fixation may allow faster initial space occupancy within an emerging community/crop. © 2011 Blackwell Publishing Ltd.

Early transduction produces highly functional chimeric antigen receptor-modified virus-specific T-cells with central memory markers: a Production Assistant for Cell Therapy (PACT) translational application

OpenAIRE

Sun, Jiali; Huye, Leslie E; Lapteva, Natalia; Mamonkin, Maksim; Hiregange, Manasa; Ballard, Brandon; Dakhova, Olga; Raghavan, Darshana; Durett, April G; Perna, Serena K; Omer, Bilal; Rollins, Lisa A; Leen, Ann M; Vera, Juan F; Dotti, Gianpietro

2015-01-01

Background Virus-specific T-cells (VSTs) proliferate exponentially after adoptive transfer into hematopoietic stem cell transplant (HSCT) recipients, eliminate virus infections, then persist and provide long-term protection from viral disease. If VSTs behaved similarly when modified with tumor-specific chimeric antigen receptors (CARs), they should have potent anti-tumor activity. This theory was evaluated by Cruz et al. in a previous clinical trial with CD19.CAR-modified VSTs, but there was ...

A thiophene-modified screen printed electrode for detection of dengue virus NS1 protein.

Science.gov (United States)

Silva, M M S; Dias, A C M S; Cordeiro, M T; Marques, E; Goulart, M O F; Dutra, R F

2014-10-01

A thiophene-modified screen printed electrode (SPE) for detection of the Dengue virus non-structural protein 1 (NS1), an important marker for acute phase diagnosis, is described. A sulfur-containing heterocyclic compound, the thiophene was incorporated to a carbon ink to prepare reproducible screen printed electrodes. After cured, the thiophene SPE was coated by gold nanoparticles conjugated to Protein A to form a nanostrutured surface. The Anti-NS1 antibodies immobilized via their Fc portions via Protein A, leaving their antigen specific sites free circumventing the problem of a random antibodies immobilization. Amperometric responses to the NS1 protein of dengue virus were obtained by cyclic voltammetries performed in presence of ferrocyanide/ferricyanide as redox probe. The calibration curve of immunosensor showed a linear response from 0.04 µg mL(-1) to 0.6 µg mL(-1) of NS1 with a good linear correlation (r=0.991, pink enhanced the electroanalytical properties of the SPEs, increasing their reproducibility and sensitivity. This point-of-care testing represents a great potential for use in epidemic situations, facilitating the early diagnosis in acute phase of dengue virus. Copyright © 2014 Elsevier B.V. All rights reserved.

Mutagenesis-mediated virus extinction: virus-dependent effect of viral load on sensitivity to lethal defection.

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Héctor Moreno

Full Text Available BACKGROUND: Lethal mutagenesis is a transition towards virus extinction mediated by enhanced mutation rates during viral genome replication, and it is currently under investigation as a potential new antiviral strategy. Viral load and virus fitness are known to influence virus extinction. Here we examine the effect or the multiplicity of infection (MOI on progeny production of several RNA viruses under enhanced mutagenesis. RESULTS: The effect of the mutagenic base analogue 5-fluorouracil (FU on the replication of the arenavirus lymphocytic choriomeningitis virus (LCMV can result either in inhibition of progeny production and virus extinction in infections carried out at low multiplicity of infection (MOI, or in a moderate titer decrease without extinction at high MOI. The effect of the MOI is similar for LCMV and vesicular stomatitis virus (VSV, but minimal or absent for the picornaviruses foot-and-mouth disease virus (FMDV and encephalomyocarditis virus (EMCV. The increase in mutation frequency and Shannon entropy (mutant spectrum complexity as a result of virus passage in the presence of FU was more accentuated at low MOI for LCMV and VSV, and at high MOI for FMDV and EMCV. We present an extension of the lethal defection model that agrees with the experimental results. CONCLUSIONS: (i Low infecting load favoured the extinction of negative strand viruses, LCMV or VSV, with an increase of mutant spectrum complexity. (ii This behaviour is not observed in RNA positive strand viruses, FMDV or EMCV. (iii The accumulation of defector genomes may underlie the MOI-dependent behaviour. (iv LCMV coinfections are allowed but superinfection is strongly restricted in BHK-21 cells. (v The dissimilar effects of the MOI on the efficiency of mutagenic-based extinction of different RNA viruses can have implications for the design of antiviral protocols based on lethal mutagenesis, presently under development.

Elevated air movement enhances stomatal sensitivity to abscisic acid in leaves developed at high relative air humidity

Directory of Open Access Journals (Sweden)

Dália R.A. Carvalho

2015-05-01

Full Text Available High relative air humidity (RH ≥ 85% during growth leads to stomata malfunctioning, resulting in water stress when plants are transferred to conditions of high evaporative demand. In this study, we hypothesized that an elevated air movement (MOV 24 h per day, during the whole period of leaf development would increase abscisic acid concentration ([ABA] enhancing stomatal functioning. Pot rose ‘Toril’ was grown at moderate (61% or high (92% RH combined with a negligible MOV or with a continuous MOV of 0.92 m s-1. High MOV reduced stomatal pore length and aperture in plants developed at high RH. Moreover, stomatal function improved when high MOV-treated plants were subjected to leaflet desiccation and ABA feeding. Endogenous concentration of ABA and its metabolites in the leaves was reduced by 35% in high RH, but contrary to our hypothesis this concentration was not significantly affected by high MOV. Interestingly, in detached leaflets grown at high RH, high MOV increased stomatal sensitivity to ABA since the amount of exogenous ABA required to decrease the transpiration rate was significantly reduced. This is the first study to show that high MOV increases stomatal functionality in leaves developed at high RH by reducing the stomatal pore length and aperture and enhancing stomatal sensitivity to ABA rather than increasing leaf [ABA].

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...... 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....

Tannic acid modified silver nanoparticles show antiviral activity in herpes simplex virus type 2 infection.

Directory of Open Access Journals (Sweden)

Piotr Orlowski

Full Text Available The interaction between silver nanoparticles and herpesviruses is attracting great interest due to their antiviral activity and possibility to use as microbicides for oral and anogenital herpes. In this work, we demonstrate that tannic acid modified silver nanoparticles sized 13 nm, 33 nm and 46 nm are capable of reducing HSV-2 infectivity both in vitro and in vivo. The antiviral activity of tannic acid modified silver nanoparticles was size-related, required direct interaction and blocked virus attachment, penetration and further spread. All tested tannic acid modified silver nanoparticles reduced both infection and inflammatory reaction in the mouse model of HSV-2 infection when used at infection or for a post-infection treatment. Smaller-sized nanoparticles induced production of cytokines and chemokines important for anti-viral response. The corresponding control buffers with tannic acid showed inferior antiviral effects in vitro and were ineffective in blocking in vivo infection. Our results show that tannic acid modified silver nanoparticles are good candidates for microbicides used in treatment of herpesvirus infections.

Curcumin is a promising inhibitor of genotype 2 porcine reproductive and respiratory syndrome virus infection.

Science.gov (United States)

Du, Taofeng; Shi, Yunpeng; Xiao, Shuqi; Li, Na; Zhao, Qin; Zhang, Angke; Nan, Yuchen; Mu, Yang; Sun, Yani; Wu, Chunyan; Zhang, Hongtao; Zhou, En-Min

2017-10-10

Porcine reproductive and respiratory syndrome virus (PRRSV) could lead to pandemic diseases and huge financial losses to the swine industry worldwide. Curcumin, a natural compound, has been reported to serve as an entry inhibitor of hepatitis C virus, chikungunya virus and vesicular stomatitis virus. In this study, we investigated the potential effect of curcumin on early stages of PRRSV infection. Curcumin inhibited infection of Marc-145 cells and porcine alveolar macrophages (PAMs) by four different genotype 2 PRRSV strains, but had no effect on the levels of major PRRSV receptor proteins on Marc-145 cells and PAMs or on PRRSV binding to Marc-145 cells. However, curcumin did block two steps of the PRRSV infection process: virus internalization and virus-mediated cell fusion. Our results suggested that an inhibition of genotype 2 PRRSV infection by curcumin is virus strain-independent, and mainly inhibited by virus internalization and cell fusion mediated by virus. Collectively, these results demonstrate that curcumin holds promise as a new anti-PRRSV drug.

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.

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.

Daya Hambat Infusum Daun Sirih Terhadap Pertumbuhan Staphylococcus aureus Yang Diisolasi Dari Denture Stomatitis ; Penelitian In Vitro.

OpenAIRE

bin Abdullah, Muhammad Naim

2011-01-01

Denture Stomatitis merupakan lesi mukosa oral berwarna merah, sakit, dan bengkak, kondisi ini karena kebiasaan jelek pada pemakai gigitiruan yang tidak mumbuka protesa pada malam hari dan jarang dibersihkan. Faktor sistemik yang mendukung terjadinya Denture Stomatitis dapat disebabkan oleh beberapa bakteri, salah satunya Staphylococcus aureus. Pencegahan Denture Stomatitis dapat dilakukan dengan sering membersihkan gigitiruan dan pemakaian obat kumur. Tujuan penelitian ini adalah untuk menguj...

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.

A genetic screen reveals Arabidopsis stomatal and/or apoplastic defenses against Pseudomonas syringae pv. tomato DC3000.

Directory of Open Access Journals (Sweden)

Weiqing Zeng

2011-10-01

Full Text Available Bacterial infection of plants often begins with colonization of the plant surface, followed by entry into the plant through wounds and natural openings (such as stomata, multiplication in the intercellular space (apoplast of the infected tissues, and dissemination of bacteria to other plants. Historically, most studies assess bacterial infection based on final outcomes of disease and/or pathogen growth using whole infected tissues; few studies have genetically distinguished the contribution of different host cell types in response to an infection. The phytotoxin coronatine (COR is produced by several pathovars of Pseudomonas syringae. COR-deficient mutants of P. s. tomato (Pst DC3000 are severely compromised in virulence, especially when inoculated onto the plant surface. We report here a genetic screen to identify Arabidopsis mutants that could rescue the virulence of COR-deficient mutant bacteria. Among the susceptible to coronatine-deficient Pst DC3000 (scord mutants were two that were defective in stomatal closure response, two that were defective in apoplast defense, and four that were defective in both stomatal and apoplast defense. Isolation of these three classes of mutants suggests that stomatal and apoplastic defenses are integrated in plants, but are genetically separable, and that COR is important for Pst DC3000 to overcome both stomatal guard cell- and apoplastic mesophyll cell-based defenses. Of the six mutants defective in bacterium-triggered stomatal closure, three are defective in salicylic acid (SA-induced stomatal closure, but exhibit normal stomatal closure in response to abscisic acid (ABA, and scord7 is compromised in both SA- and ABA-induced stomatal closure. We have cloned SCORD3, which is required for salicylic acid (SA biosynthesis, and SCORD5, which encodes an ATP-binding cassette (ABC protein, AtGCN20/AtABCF3, predicted to be involved in stress-associated protein translation control. Identification of SCORD5 begins to

Ozone slows stomatal response to light and leaf wounding in a Mediterranean evergreen broadleaf, Arbutus unedo

Energy Technology Data Exchange (ETDEWEB)

Paoletti, Elena [Istituto Protezione Piante, Consiglio Nazionale delle Ricerche, Via Madonna del Piano, I-50019 Sesto Fiorentino (Italy)]. E-mail: e.paoletti@ipp.cnr.it

2005-04-01

The effect of a 90-d ozone exposure (charcoal-filtered air or 110 nmol mol{sup -1} O{sub 3}) on stomatal conductance (g{sub s}) was investigated in the Mediterranean evergreen broadleaf Arbutus unedo L. Ozone did not significantly reduce midday steady-state g{sub s} compared to controls. However, it slowed stomatal response to abrupt reduction of light intensity and to increasing water stress, applied by severing the leaf midrib. Ozone slowed stomatal closure, rather than aperture. Nevertheless, vein-cutting did not allow ozonated leaves to reach the pre-injury g{sub s} levels, like controls did, suggesting re-opening was still, slowly in progress. The sluggish behaviour was recorded 10 days after cessation of O{sub 3} exposure ('memory effect') and may affect stomatal control in response to sunflecks and leaf wounding. Mediterranean evergreen broadleaves are regarded as tolerant to O{sub 3} exposure. Nevertheless, measurements of steady-state g{sub s} at midday may not account for altered stomatal responses to stressors. - In response to ozone exposure, stomata were slower in closing rather than in opening.

Ozone slows stomatal response to light and leaf wounding in a Mediterranean evergreen broadleaf, Arbutus unedo

International Nuclear Information System (INIS)

Paoletti, Elena

2005-01-01

The effect of a 90-d ozone exposure (charcoal-filtered air or 110 nmol mol -1 O 3 ) on stomatal conductance (g s ) was investigated in the Mediterranean evergreen broadleaf Arbutus unedo L. Ozone did not significantly reduce midday steady-state g s compared to controls. However, it slowed stomatal response to abrupt reduction of light intensity and to increasing water stress, applied by severing the leaf midrib. Ozone slowed stomatal closure, rather than aperture. Nevertheless, vein-cutting did not allow ozonated leaves to reach the pre-injury g s levels, like controls did, suggesting re-opening was still, slowly in progress. The sluggish behaviour was recorded 10 days after cessation of O 3 exposure ('memory effect') and may affect stomatal control in response to sunflecks and leaf wounding. Mediterranean evergreen broadleaves are regarded as tolerant to O 3 exposure. Nevertheless, measurements of steady-state g s at midday may not account for altered stomatal responses to stressors. - In response to ozone exposure, stomata were slower in closing rather than in opening

ABA-Induced Stomatal Closure Involves ALMT4, a Phosphorylation-Dependent Vacuolar Anion Channel of Arabidopsis[OPEN

Science.gov (United States)

Baetz, Ulrike; Huck, Nicola V.; Zhang, Jingbo

2017-01-01

Stomatal pores are formed between a pair of guard cells and allow plant uptake of CO2 and water evaporation. Their aperture depends on changes in osmolyte concentration of guard cell vacuoles, specifically of K+ and Mal2−. Efflux of Mal2− from the vacuole is required for stomatal closure; however, it is not clear how the anion is released. Here, we report the identification of ALMT4 (ALUMINUM ACTIVATED MALATE TRANSPORTER4) as an Arabidopsis thaliana ion channel that can mediate Mal2− release from the vacuole and is required for stomatal closure in response to abscisic acid (ABA). Knockout mutants showed impaired stomatal closure in response to the drought stress hormone ABA and increased whole-plant wilting in response to drought and ABA. Electrophysiological data show that ALMT4 can mediate Mal2− efflux and that the channel activity is dependent on a phosphorylatable C-terminal serine. Dephosphomimetic mutants of ALMT4 S382 showed increased channel activity and Mal2− efflux. Reconstituting the active channel in almt4 mutants impaired growth and stomatal opening. Phosphomimetic mutants were electrically inactive and phenocopied the almt4 mutants. Surprisingly, S382 can be phosphorylated by mitogen-activated protein kinases in vitro. In brief, ALMT4 likely mediates Mal2− efflux during ABA-induced stomatal closure and its activity depends on phosphorylation. PMID:28874508

Evidence-based modelling of diverse plant water use strategies on stomatal and non-stomatal components under drought

Science.gov (United States)

zhou, S.; Prentice, C.; Medlyn, B. E.; Sabaté, S.

2013-12-01

Models disagree on how to represent effects of drought stress on plant gas exchange. Some models assume drought stress affects the marginal water use efficiency of plants (marginal WUE; i.e. the change in photosynthesis per unit of change in transpiration) whereas others assume drought stress acts directly on photosynthetic capacity. It is not clear whether either of these approaches is sufficient to capture the drought response, or whether the effect of drought varies among species and functional types. A collection of Eucalyptus and Quercus species derived from different hydro-climate habitats, in together with two European riparian species, were conducted with drought treatments respectively in Australia and Spain for three months. Measurements included net CO2 assimilation rate versus substomatal CO2 concentration (A-Ci) curves, fluorescence, and predawn leaf water potential at increasing levels of water stress. The correlations with quantitative plant traits of leaf, stomata, vessel, and wood density, leaf nitrogen content and 13C discrimination were also explored. We analysed the effect of drought effect on leaf gas exchange with a recently developed stomatal model that reconciles the empirical and optimal approaches on predicting optimal stomatal conductance. The model's single parameter g1 is a decreasing function of marginal WUE. The two genera showed consistence on the contrasting response patterns between species derived from mesic and arid habitats, which differed greatly in their estimated g1 values under moist conditions, and in the rate at which g1 declined with water stress. They also differed greatly in the predawn water potential at which apparent carboxylation capacity (apparent Vcmax) and mesophyll conductance (gm) declined most steeply, and in the steepness of this decline. Principal components analysis revealed a gradient in water relation strategies from sclerophyll species to malacophyll species. Malacophylls had higher g1, apparent Vcmax

Development and Characterization of a Multiplexed RT-PCR Species Specific Assay for Bovine and one for Porcine Foot-and-Mouth Disease Virus Rule-Out Supplemental Materials

Energy Technology Data Exchange (ETDEWEB)

Smith, S; Danganan, L; Tammero, L; Lenhoff, R; Naraghi-arani, P; Hindson, B

2007-08-06

Lawrence Livermore National Laboratory (LLNL), in collaboration with the Department of Homeland Security (DHS) and the United States Department of Agriculture (USDA), Animal and Plant Health Inspection Services (APHIS) has developed advanced rapid diagnostics that may be used within the National Animal Health Laboratory Network (NAHLN), the National Veterinary Services Laboratory (Ames, Iowa) and the Plum Island Animal Disease Center (PIADC). This effort has the potential to improve our nation's ability to discriminate between foreign animal diseases and those that are endemic using a single assay, thereby increasing our ability to protect animal populations of high economic importance in the United States. Under 2005 DHS funding we have developed multiplexed (MUX) nucleic-acid-based PCR assays that combine foot-and-mouth disease virus (FMDV) detection with rule-out tests for two other foreign animal diseases Vesicular Exanthema of Swine (VESV) and Swine Vesicular Disease (SVD) and four other domestic viral diseases Bovine Viral Diarrhea Virus (BVDV), Bovine Herpes Virus 1 (BHV-1 or Infectious Bovine Rhinotracheitus IBR), Bluetongue virus (BTV) and Parapox virus complex (which includes Bovine Papular Stomatitis Virus BPSV, Orf of sheep, and Pseudocowpox). Under 2006 funding we have developed a Multiplexed PCR [MUX] porcine assay for detection of FMDV with rule out tests for VESV and SVD foreign animal diseases in addition to one other domestic vesicular animal disease vesicular stomatitis virus (VSV) and one domestic animal disease of swine porcine reproductive and respiratory syndrome (PRRS). We have also developed a MUX bovine assay for detection of FMDV with rule out tests for the two bovine foreign animal diseases malignant catarrhal fever (MCF), rinderpest virus (RPV) and the domestic diseases vesicular stomatitis virus (VSV), bovine viral diarrhea virus (BVDV), infectious bovine rhinotracheitus virus (BHV-1), bluetongue virus (BTV), and the Parapox

Observations of leaf stomatal conductance at the canopy scale: an atmospheric modeling perspective

International Nuclear Information System (INIS)

Avissar, R.

1993-01-01

Plant stomata play a key role in the redistribution of energy received on vegetated land into sensible and latent heat. As a result, they have a considerable impact on the atmospheric planetary boundary layer, the hydrologic cycle, the climate, and the weather. Current parameterizations of the stomatal mechanism in state-of-the-art atmospheric models are based on empirical relations that are established at the leaf scale between stomatal conductance and environmental conditions. In order to evaluate these parameterizations, an experiment was carried out on a potato field in New Jersey during the summer of 1989. Stomatal conductances were measured within a small homogeneous area in the middle of the potato field and under a relatively broad range of atmospheric conditions. A large variability of stomatal conductances was observed. This variability, which was associated with the variability of micro-environmental and physiological conditions that is found even in a homogeneous canopy, cannot be simulated explicitly on the scale of a single agricultural field and,a fortiori, on the scale of atmospheric models. Furthermore, this variability could not be related to the environmental conditions measured at a height of 2 m above the plant canopy simultaneously with the conductances, reinforcing the concept of scale decoupling suggested by Jarvis and McNaughton (1986) and McNaughton and Jarvis (1991). Thus, for atmospheric modeling purposes, a parameterization of stomatal conductance at the canopy scale using external environmental forcing conditions seems more appropriate than a parameterization based on leaf-scale stomatal conductance, as currently adopted in state-of-the-art atmospheric models. The measured variability was characterized by a lognormal probability density function (pdf) that remained relatively stable during the entire measuring period. These observations support conclusions by McNaughton and Jarvis (1991) that, unlike current parameterizations, a

Observations of leaf stomatal conductance at the canopy scale: an atmospheric modeling perspective

International Nuclear Information System (INIS)

Avissar, R.

1993-01-01

Plant stomata play a key role in the redistribution of energy received on vegetated land into sensible and latent heat. As a result, they have a considerable impact on the atmospheric planetary boundary layer, the hydrologic cycle, the climate, and the weather. Current parameterizations of the stomatal mechanism in state-of-the-art atmospheric models are based on empirical relations that are established at the leaf scale between stomatal conductance and environmental conditions. In order to evaluate these parameterizations, an experiment was carried out on a potato field in New Jersey during the summer of 1989. Stomatal conductances were measured within a small homogeneous area in the middle of the potato field and under a relatively broad range of atmospheric conditions. A large variability of stomatal conductances was observed. This variability, which was associated with the variability of micro-environmental and physiological conditions that is found even in a homogeneous canopy, cannot be simulated explicitly on the scale of a single agricultural field and, a fortiori, on the scale of atmospheric models. Furthermore, this variability could not be related to the environmental conditions measured at a height of 2 m above the plant canopy simultaneously with the conductances, reinforcing the concept of scale decoupling suggested by Jarvis and McNaughton (1986) and McNaughton and Jarvis (1991). Thus, for atmospheric modeling purposes, a parameterization of stomatal conductance at the canopy scale using external environmental forcing conditions seems more appropriate than a parameterization based on leaf-scale stomatal conductance, as currently adopted in state-of-the-art atmospheric models. The measured variability was characterized by a lognormal probability density function (pdf) that remained relatively stable during the entire measuring period. These observations support conclusions by McNaughton and Jarvis (1991) that, unlike current parameterizations, a

Thermal infrared imaging of the temporal variability in stomatal conductance for fruit trees

Science.gov (United States)

Struthers, Raymond; Ivanova, Anna; Tits, Laurent; Swennen, Rony; Coppin, Pol

2015-07-01

Repeated measurements using thermal infrared remote sensing were used to characterize the change in canopy temperature over time and factors that influenced this change on 'Conference' pear trees (Pyrus communis L.). Three different types of sensors were used, a leaf porometer to measure leaf stomatal conductance, a thermal infrared camera to measure the canopy temperature and a meteorological sensor to measure weather variables. Stomatal conductance of water stressed pear was significantly lower than in the control group 9 days after stress began. This decrease in stomatal conductance reduced transpiration, reducing evaporative cooling that increased canopy temperature. Using thermal infrared imaging with wavelengths between 7.5 and13 μm, the first significant difference was measured 18 days after stress began. A second order derivative described the average rate of change of the difference between the stress treatment and control group. The average rate of change for stomatal conductance was 0.06 (mmol m-2 s-1) and for canopy temperature was -0.04 (°C) with respect to days. Thermal infrared remote sensing and data analysis presented in this study demonstrated that the differences in canopy temperatures between the water stress and control treatment due to stomata regulation can be validated.

Simultaneous requirement of carbon dioxide and abscisic acid for stomatal closing in Xanthium strumarium L.

Science.gov (United States)

Raschke, K

1975-01-01

Open stomata of detached leaves of Xanthium strumarium L. closed only when carbon dioxide and abscisic acid (ABA) were presented simultaneously. Three parameters of stomatal closing were determined after additions of ABA to the irrigation water of detached leaves, while the leaves were exposed to various CO2 concentrations ([CO2]s) in the air; a) the delay between addition of ABA and a reduction of stomatal conductance by 5%, b) the velocity of stomatal closing, and c) the new conductance. Changes in all three parameters showed that stomatal responses to ABA were enhanced by CO2; this effect followed saturation kinetics. Half saturation occurred at an estimated [CO2] in the stomatal pore of 200 μl l(-1). With respect to ABA, stomata responded in normal air with half their maximal amplitude at [ABA]s between 10(-6) and 10(-5) M(+-)-ABA. The amounts of ABA taken up by the leaves during the delay increased with a power strumarium.Based on earlier findings and on the results of this investigation it is suggested that stomata close if the cytoplasm of the guard cells contains much malate and H(+). The acid content in turn is determined by the relative rates of production of malic acid (from endogenous as well as exogenous CO2) and its removal (by transport of the anion into the vacuole and exchange of the H(+) for K(+) with the environment of the guard cells). The simultaneous requirement of CO2 and ABA for stomatal closure leads to the inference that ABA inhibits the expulsion of H(+) from guard cells.

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...

Does Toll-like receptor 3 play a biological role in virus infections?

International Nuclear Information System (INIS)

Edelmann, Kurt H.; Richardson-Burns, Sarah; Alexopoulou, Lena; Tyler, Kenneth L.; Flavell, Richard A.; Oldstone, Michael B.A.

2004-01-01

The Toll-like receptor (TLR) family functions to recognize conserved microbial and viral structures with the purpose of activating signal pathways to instigate immune responses against infections by these organisms. For example, in vitro studies reveal that the TLR3 ligand is a double-stranded RNA (dsRNA), a product of viral infections. From this observation, it has been proposed that TLR3 is likely an important first signal for virus infections. We approached this issue by investigating the role of TLR3 in four different infectious viral models (lymphocytic choriomeningitis virus (LCMV), vesicular stomatitis virus (VSV), murine cytomegalovirus (MCMV), and reovirus) and in TLR3 genetically deficient ( -/- ) mice. Our results indicate that TLR3 is not universally required for the generation of effective antiviral responses because the absence of TLR3 does not alter either viral pathogenesis or impair host's generation of adaptive antiviral responses to these viruses

Sensitivity analysis of a parameterization of the stomatal component of the DO3SE model for Quercus ilex to estimate ozone fluxes

International Nuclear Information System (INIS)

Alonso, Rocio; Elvira, Susana; Sanz, Maria J.; Gerosa, Giacomo; Emberson, Lisa D.; Bermejo, Victoria; Gimeno, Benjamin S.

2008-01-01

A sensitivity analysis of a proposed parameterization of the stomatal conductance (g s ) module of the European ozone deposition model (DO 3 SE) for Quercus ilex was performed. The performance of the model was tested against measured g s in the field at three sites in Spain. The best fit of the model was found for those sites, or during those periods, facing no or mild stress conditions, but a worse performance was found under severe drought or temperature stress, mostly occurring at continental sites. The best performance was obtained when both f phen and f SWP were included. A local parameterization accounting for the lower temperatures recorded in winter and the higher water shortage at the continental sites resulted in a better performance of the model. The overall results indicate that two different parameterizations of the model are needed, one for marine-influenced sites and another one for continental sites. - No redundancy between phenological and water-related modifying functions was found when estimating stomatal behavior of Holm oak

Aquaporins Contribute to ABA-Triggered Stomatal Closure through OST1-Mediated Phosphorylation

Science.gov (United States)

Grondin, Alexandre; Rodrigues, Olivier; Verdoucq, Lionel; Merlot, Sylvain; Leonhardt, Nathalie; Maurel, Christophe

2015-01-01

Stomatal movements in response to environmental stimuli critically control the plant water status. Although these movements are governed by osmotically driven changes in guard cell volume, the role of membrane water channels (aquaporins) has remained hypothetical. Assays in epidermal peels showed that knockout Arabidopsis thaliana plants lacking the Plasma membrane Intrinsic Protein 2;1 (PIP2;1) aquaporin have a defect in stomatal closure, specifically in response to abscisic acid (ABA). ABA induced a 2-fold increase in osmotic water permeability (Pf) of guard cell protoplasts and an accumulation of reactive oxygen species in guard cells, which were both abrogated in pip2;1 plants. Open stomata 1 (OST1)/Snf1-related protein kinase 2.6 (SnRK2.6), a protein kinase involved in guard cell ABA signaling, was able to phosphorylate a cytosolic PIP2;1 peptide at Ser-121. OST1 enhanced PIP2;1 water transport activity when coexpressed in Xenopus laevis oocytes. Upon expression in pip2;1 plants, a phosphomimetic form (Ser121Asp) but not a phosphodeficient form (Ser121Ala) of PIP2;1 constitutively enhanced the Pf of guard cell protoplasts while suppressing its ABA-dependent activation and was able to restore ABA-dependent stomatal closure in pip2;1. This work supports a model whereby ABA-triggered stomatal closure requires an increase in guard cell permeability to water and possibly hydrogen peroxide, through OST1-dependent phosphorylation of PIP2;1 at Ser-121. PMID:26163575

Phytomelatonin receptor PMTR1-mediated signaling regulates stomatal closure in Arabidopsis thaliana.

Science.gov (United States)

Wei, Jian; Li, Dong-Xu; Zhang, Jia-Rong; Shan, Chi; Rengel, Zed; Song, Zhong-Bang; Chen, Qi

2018-04-27

Melatonin has been detected in plants in 1995; however, the function and signaling pathway of this putative phytohormone are largely undetermined due to a lack of knowledge about its receptor. Here, we discovered the first phytomelatonin receptor (CAND2/PMTR1) in Arabidopsis thaliana and found that melatonin governs the receptor-dependent stomatal closure. The application of melatonin induced stomatal closure through the heterotrimeric G protein α subunit-regulated H 2 O 2 and Ca 2+ signals. The Arabidopsis mutant lines lacking AtCand2 that encodes a candidate G protein-coupled receptor were insensitive to melatonin-induced stomatal closure. Accordingly, the melatonin-induced H 2 O 2 production and Ca 2+ influx were completely abolished in cand2. CAND2 is a membrane protein that interacts with GPA1 and the expression of AtCand2 was tightly regulated by melatonin in various organs and guard cells. CAND2 showed saturable and specific 125 I-melatonin binding, with apparent K d (dissociation constant) of 0.73 ± 0.10 nmol/L (r 2  = .99), demonstrating this protein is a phytomelatonin receptor (PMTR1). Our results suggest that the phytomelatonin regulation of stomatal closure is dependent on its receptor CAND2/PMTR1-mediated H 2 O 2 and Ca 2+ signaling transduction cascade. © 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

The Arabidopsis lectin receptor kinase LecRK-V.5 represses stomatal immunity induced by Pseudomonas syringae pv. tomato DC3000.

Directory of Open Access Journals (Sweden)

Marie Desclos-Theveniau

2012-02-01

Full Text Available Stomata play an important role in plant innate immunity by limiting pathogen entry into leaves but molecular mechanisms regulating stomatal closure upon pathogen perception are not well understood. Here we show that the Arabidopsis thaliana L-type lectin receptor kinase-V.5 (LecRK-V.5 negatively regulates stomatal immunity. Loss of LecRK-V.5 function increased resistance to surface inoculation with virulent bacteria Pseudomonas syringae pv tomato DC3000. Levels of resistance were not affected after infiltration-inoculation, suggesting that LecRK-V.5 functions at an early defense stage. By contrast, lines overexpressing LecRK-V.5 were more susceptible to Pst DC3000. Enhanced resistance in lecrk-V.5 mutants was correlated with constitutive stomatal closure, while increased susceptibility phenotypes in overexpression lines were associated with early stomatal reopening. Lines overexpressing LecRK-V.5 also demonstrated a defective stomatal closure after pathogen-associated molecular pattern (PAMP treatments. LecRK-V.5 is rapidly expressed in stomatal guard cells after bacterial inoculation or treatment with the bacterial PAMP flagellin. In addition, lecrk-V.5 mutants guard cells exhibited constitutive accumulation of reactive oxygen species (ROS and inhibition of ROS production opened stomata of lecrk-V.5. LecRK-V.5 is also shown to interfere with abscisic acid-mediated stomatal closure signaling upstream of ROS production. These results provide genetic evidences that LecRK-V.5 negatively regulates stomatal immunity upstream of ROS biosynthesis. Our data reveal that plants have evolved mechanisms to reverse bacteria-mediated stomatal closure to prevent long-term effect on CO(2 uptake and photosynthesis.

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-nine pat......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...... analyzed and compared between groups. Nineteen patients had OLP, primarily with ulcerative lesions on the buccal mucosa, 19 patients had OLL, and 11 patients had generalized stomatitis. All patients had oral symptoms, mainly stinging and burning. Nineteen patients and 10 healthy subjects had contact...... higher levels of IL-6 than the healthy subjects. Interferon-γ, IL-12p40, and IL-12p70 were below detection limit. Our findings indicate that OLP, OLL, and generalized stomatitis cannot be discriminated by means of the selected serum cytokines, and that the presence of concomitant contact allergy does...

Evaluation of the stomatal conductance formulation in the EMEP ozone deposition model for Picea abies

Science.gov (United States)

Wieser, G.; Emberson, L. D.

It is widely acknowledged that the possible impacts of ozone on forest trees are more closely related to ozone flux through the stomata than to external ozone exposure. However, the application of the flux approach on a European scale requires the availability of appropriate models, such as the European Monitoring and Evaluation Programme (EMEP) ozone deposition model, for estimating ozone flux and cumulative ozone uptake. Within this model stomatal conductance is the key variable, since it determines the amount of ozone absorbed by the leaves. This paper describes the suitability of the existing EMEP ozone deposition model parameterisation and formulation to represent stomatal behaviour determined from field measurements on adult Norway spruce ( Picea abies (L.) Karst.) trees in the Central European Alps. Parameters affecting maximum stomatal conductance (e.g. seasonal phenology, needle position, needle age, nutrient deficiency and ozone itself) and stomatal response functions to temperature, irradiance, vapour pressure deficit, and soil water content are investigated. Finally, current limitations and possible alterations of the EMEP model will be discussed with respect to spatial scales of available input data for future flux modelling.

Modeling stomatal conductance in the earth system: linking leaf water-use efficiency and water transport along the soil-plant-atmosphere continuum

Science.gov (United States)

Bonan, G. B.; Williams, M.; Fisher, R. A.; Oleson, K. W.

2014-09-01

The Ball-Berry stomatal conductance model is commonly used in earth system models to simulate biotic regulation of evapotranspiration. However, the dependence of stomatal conductance (gs) on vapor pressure deficit (Ds) and soil moisture must be empirically parameterized. We evaluated the Ball-Berry model used in the Community Land Model version 4.5 (CLM4.5) and an alternative stomatal conductance model that links leaf gas exchange, plant hydraulic constraints, and the soil-plant-atmosphere continuum (SPA). The SPA model simulates stomatal conductance numerically by (1) optimizing photosynthetic carbon gain per unit water loss while (2) constraining stomatal opening to prevent leaf water potential from dropping below a critical minimum. We evaluated two optimization algorithms: intrinsic water-use efficiency (ΔAn /Δgs, the marginal carbon gain of stomatal opening) and water-use efficiency (ΔAn /ΔEl, the marginal carbon gain of transpiration water loss). We implemented the stomatal models in a multi-layer plant canopy model to resolve profiles of gas exchange, leaf water potential, and plant hydraulics within the canopy, and evaluated the simulations using leaf analyses, eddy covariance fluxes at six forest sites, and parameter sensitivity analyses. The primary differences among stomatal models relate to soil moisture stress and vapor pressure deficit responses. Without soil moisture stress, the performance of the SPA stomatal model was comparable to or slightly better than the CLM Ball-Berry model in flux tower simulations, but was significantly better than the CLM Ball-Berry model when there was soil moisture stress. Functional dependence of gs on soil moisture emerged from water flow along the soil-to-leaf pathway rather than being imposed a priori, as in the CLM Ball-Berry model. Similar functional dependence of gs on Ds emerged from the ΔAn/ΔEl optimization, but not the ΔAn /gs optimization. Two parameters (stomatal efficiency and root hydraulic

Overexpression of plasma membrane H+-ATPase in guard cells promotes light-induced stomatal opening and enhances plant growth.

Science.gov (United States)

Wang, Yin; Noguchi, Ko; Ono, Natsuko; Inoue, Shin-ichiro; Terashima, Ichiro; Kinoshita, Toshinori

2014-01-07

Stomatal pores surrounded by a pair of guard cells in the plant epidermis control gas exchange between plants and the atmosphere in response to light, CO2, and the plant hormone abscisic acid. Light-induced stomatal opening is mediated by at least three key components: the blue light receptor phototropin (phot1 and phot2), plasma membrane H(+)-ATPase, and plasma membrane inward-rectifying K(+) channels. Very few attempts have been made to enhance stomatal opening with the goal of increasing photosynthesis and plant growth, even though stomatal resistance is thought to be the major limiting factor for CO2 uptake by plants. Here, we show that transgenic Arabidopsis plants overexpressing H(+)-ATPase using the strong guard cell promoter GC1 showed enhanced light-induced stomatal opening, photosynthesis, and plant growth. The transgenic plants produced larger and increased numbers of rosette leaves, with ∼42-63% greater fresh and dry weights than the wild type in the first 25 d of growth. The dry weights of total flowering stems of 45-d-old transgenic plants, including seeds, siliques, and flowers, were ∼36-41% greater than those of the wild type. In addition, stomata in the transgenic plants closed normally in response to darkness and abscisic acid. In contrast, the overexpression of phototropin or inward-rectifying K(+) channels in guard cells had no effect on these phenotypes. These results demonstrate that stomatal aperture is a limiting factor in photosynthesis and plant growth, and that manipulation of stomatal opening by overexpressing H(+)-ATPase in guard cells is useful for the promotion of plant growth.

Impact of Canopy Coupling on Canopy Average Stomatal Conductance Across Seven Tree Species in Northern Wisconsin

Science.gov (United States)

Ewers, B. E.; Mackay, D. S.; Samanta, S.; Ahl, D. E.; Burrows, S. S.; Gower, S. T.

2001-12-01

Land use changes over the last century in northern Wisconsin have resulted in a heterogeneous landscape composed of the following four main forest types: northern hardwoods, northern conifer, aspen/fir, and forested wetland. Based on sap flux measurements, aspen/fir has twice the canopy transpiration of northern hardwoods. In addition, daily transpiration was only explained by daily average vapor pressure deficit across the cover types. The objective of this study was to determine if canopy average stomatal conductance could be used to explain the species effects on tree transpiration. Our first hypothesis is that across all of the species, stomatal conductance will respond to vapor pressure deficit so as to maintain a minimum leaf water potential to prevent catostrophic cavitiation. The consequence of this hypothesis is that among species and individuals there is a proportionality between high stomatal conductance and the sensitivity of stomatal conductance to vapor pressure deficit. Our second hypothesis is that species that do not follow the proportionality deviate because the canopies are decoupled from the atmosphere. To test our two hypotheses we calculated canopy average stomatal conductance from sap flux measurements using an inversion of the Penman-Monteith equation. We estimated the canopy coupling using a leaf energy budget model that requires leaf transpiration and canopy aerodynamic conductance. We optimized the parameters of the aerodynamic conductance model using a Monte Carlo technique across six parameters. We determined the optimal model for each species by selecting parameter sets that resulted in the proportionality of our first hypothesis. We then tested the optimal energy budget models of each species by comparing leaf temperature and leaf width predicted by the models to measurements of each tree species. In red pine, sugar maple, and trembling aspen trees under high canopy coupling conditions, we found the hypothesized proportionality

Ebolavirus Vaccines: Progress in the Fight Against Ebola Virus Disease

Directory of Open Access Journals (Sweden)

Xiao-Xin Wu

2015-11-01

Full Text Available Ebolaviruses are highly infectious pathogens that cause lethal Ebola virus disease (EVD in humans and non-human primates (NHPs. Due to their high pathogenicity and transmissibility, as well as the potential to be misused as a bioterrorism agent, ebolaviruses would threaten the health of global populations if not controlled. In this review, we describe the origin and structure of ebolaviruses and the development of vaccines from the beginning of the 1980s, including conventional ebolavirus vaccines, DNA vaccines, Ebola virus-like particles (VLPs, vaccinia virus-based vaccines, Venezuelan equine encephalitis virus (VEEV-like replicon particles, Kunjin virus-based vaccine, recombinant Zaire Ebolavirus∆VP30, recombinant cytomegalovirus (CMV-based vaccines, recombinant rabies virus (RABV-based vaccines, recombinant paramyxovirus-based vaccines, adenovirus-based vaccines and vesicular stomatitis virus (VSV-based vaccines. No licensed vaccine or specific treatment is currently available to counteract ebolavirus infection, although DNA plasmids and several viral vector approaches have been evaluated as promising vaccine platforms. These vaccine candidates have been confirmed to be successful in protecting NHPs against lethal infection. Moreover, these vaccine candidates were successfully advanced to clinical trials. The present review provides an update of the current research on Ebola vaccines, with the aim of providing an overview on current prospects in the fight against EVD.

Sensitivity analysis of a parameterization of the stomatal component of the DO{sub 3}SE model for Quercus ilex to estimate ozone fluxes

Energy Technology Data Exchange (ETDEWEB)

Alonso, Rocio [Ecotoxicology of Air Pollution, CIEMAT, Avenida Complutense 22, 28040 Madrid (Spain)], E-mail: rocio.alonso@ciemat.es; Elvira, Susana [Ecotoxicology of Air Pollution, CIEMAT, Avenida Complutense 22, 28040 Madrid (Spain)], E-mail: susana.elvira@ciemat.es; Sanz, Maria J. [Fundacion CEAM, Charles Darwin 14, 46980 Paterna, Valencia (Spain)], E-mail: mjose@ceam.es; Gerosa, Giacomo [Department of Mathematics and Physics, Universita Cattolica del Sacro Cuore, via Musei 41, 25121 Brescia (Italy)], E-mail: giacomo.gerosa@unicatt.it; Emberson, Lisa D. [Stockholm Environment Institute, University of York, York YO 10 5DD (United Kingdom)], E-mail: lde1@york.ac.uk; Bermejo, Victoria [Ecotoxicology of Air Pollution, CIEMAT, Avenida Complutense 22, 28040 Madrid (Spain)], E-mail: victoria.bermejo@ciemat.es; Gimeno, Benjamin S. [Ecotoxicology of Air Pollution, CIEMAT, Avenida Complutense 22, 28040 Madrid (Spain)], E-mail: benjamin.gimeno@ciemat.es

2008-10-15

A sensitivity analysis of a proposed parameterization of the stomatal conductance (g{sub s}) module of the European ozone deposition model (DO{sub 3}SE) for Quercus ilex was performed. The performance of the model was tested against measured g{sub s} in the field at three sites in Spain. The best fit of the model was found for those sites, or during those periods, facing no or mild stress conditions, but a worse performance was found under severe drought or temperature stress, mostly occurring at continental sites. The best performance was obtained when both f{sub phen} and f{sub SWP} were included. A local parameterization accounting for the lower temperatures recorded in winter and the higher water shortage at the continental sites resulted in a better performance of the model. The overall results indicate that two different parameterizations of the model are needed, one for marine-influenced sites and another one for continental sites. - No redundancy between phenological and water-related modifying functions was found when estimating stomatal behavior of Holm oak.

The glycoprotein genes and gene junctions of the fish rhabdoviruses spring viremia of carp virus and hirame rhabdovirus: Analysis of relationships with other rhabdoviruses

Science.gov (United States)

Bjorklund, H.V.; Higman, K.H.; Kurath, G.

1996-01-01

The nucleotide sequences of the glycoprotein genes and all of the internal gene junctions of the fish pathogenic rhabdoviruses spring viremia of carp virus (SVCV) and hirame rhabdovirus (HIRRV) have been determined from cDNA clones generated from viral genomic RNA. The SVCV glycoprotein gene sequence is 1588 nucleotides (nt) long and encodes a 509 amino acid (aa) protein. The HIRRV glycoprotein gene sequence comprises 1612 nt, coding for a 508 aa protein. In sequence comparisons of 15 rhabdovirus glycoproteins, the SVCV glycoprotein gene showed the highest amino acid sequence identity (31.2–33.2%) with vesicular stomatitis New Jersey virus (VSNJV), Chandipura virus (CHPV) and vesicular stomatitis Indiana virus (VSIV). The HIRRV glycoprotein gene showed a very high amino acid sequence identity (74.3%) with the glycoprotein gene of another fish pathogenic rhabdovirus, infectious hematopoietic necrosis virus (IHNV), but no significant similarity with glycoproteins of VSIV or rabies virus (RABV). In phylogenetic analyses SVCV was grouped consistently with VSIV, VSNJV and CHPV in the Vesiculovirus genus of Rhabdoviridae. The fish rhabdoviruses HIRRV, IHNV and viral hemorrhagic septicemia virus (VHSV) showed close relationships with each other, but only very distant relationships with mammalian rhabdoviruses. The gene junctions are highly conserved between SVCV and VSIV, well conserved between IHNV and HIRRV, but not conserved between HIRRV/IHNV and RABV. Based on the combined results we suggest that the fish lyssa-type rhabdoviruses HIRRV, IHNV and VHSV may be grouped in their own genus within the family Rhabdoviridae. Aquarhabdovirus has been proposed for the name of this new genus.

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...

Stomatal development in barley as a bioassay for cell differentation: its use with X-rays and gibberellic acid

Energy Technology Data Exchange (ETDEWEB)

Zeiger, E; Rafalowsky, J [Chile Univ., Santiago. Departamento de Biologia y Genetica

1976-01-01

A bioassay for cell differentiation during stomatal development in barley (Hordeum vulgare L.) has been defined. It uses cell kinetics analysis to follow the temporal course of cell divisions in the developmental sequence. The rate of displacement of the divisions along the stomatal rows provides a measure of differentiation. Physical factors affecting differentiation may be tested with intact seedlings. The bioassay showed that X-ray irradiation inhibited the divisions leading to stomatal formation. The inhibition kinetics was similar to the one observed in root meristems. Chemical substances are tested by culturing excised shoots in a synthetic medium. Detached leaves responded to sucrose and light with increasing rates of stomatal divisions. Gibberellic acid (GA/sub 3/) was assayed for its effects on the growth of the leaf and the differentiation of stomata. GA/sub 3/ increased the overall length of the leaves without affecting the rates of cell division. The treated cells responded with increased elongation rates and a precocious initiation and completion of cell enlargement. GA/sub 3/ had no specific effect on stomatal differentiation.

Sensitivity of stomatal conductance to soil moisture: implications for tropospheric ozone

Directory of Open Access Journals (Sweden)

A. Anav

2018-04-01

Full Text Available Soil moisture and water stress play a pivotal role in regulating stomatal behaviour of plants; however, in the last decade, the role of water availability has often been neglected in atmospheric chemistry modelling studies as well as in integrated risk assessments, despite the fact that plants remove a large amount of atmospheric compounds from the lower troposphere through stomata. The main aim of this study is to evaluate, within the chemistry transport model CHIMERE, the effect of soil water limitation on stomatal conductance and assess the resulting changes in atmospheric chemistry testing various hypotheses of water uptake by plants in the rooting zone. Results highlight how dry deposition significantly declines when soil moisture is used to regulate the stomatal opening, mainly in the semi-arid environments: in particular, over Europe the amount of ozone removed by dry deposition in one year without considering any soil water limitation to stomatal conductance is about 8.5 TgO3, while using a dynamic layer that ensures that plants maximize the water uptake from soil, we found a reduction of about 10 % in the amount of ozone removed by dry deposition ( ∼  7.7 TgO3. Although dry deposition occurs from the top of canopy to ground level, it affects the concentration of gases remaining in the lower atmosphere, with a significant impact on ozone concentration (up to 4 ppb extending from the surface to the upper troposphere (up to 650 hPa. Our results shed light on the importance of improving the parameterizations of processes occurring at plant level (i.e. from the soil to the canopy as they have significant implications for concentration of gases in the lower troposphere and resulting risk assessments for vegetation or human health.

Sensitivity of stomatal conductance to soil moisture: implications for tropospheric ozone

Science.gov (United States)

Anav, Alessandro; Proietti, Chiara; Menut, Laurent; Carnicelli, Stefano; De Marco, Alessandra; Paoletti, Elena

2018-04-01

Soil moisture and water stress play a pivotal role in regulating stomatal behaviour of plants; however, in the last decade, the role of water availability has often been neglected in atmospheric chemistry modelling studies as well as in integrated risk assessments, despite the fact that plants remove a large amount of atmospheric compounds from the lower troposphere through stomata. The main aim of this study is to evaluate, within the chemistry transport model CHIMERE, the effect of soil water limitation on stomatal conductance and assess the resulting changes in atmospheric chemistry testing various hypotheses of water uptake by plants in the rooting zone. Results highlight how dry deposition significantly declines when soil moisture is used to regulate the stomatal opening, mainly in the semi-arid environments: in particular, over Europe the amount of ozone removed by dry deposition in one year without considering any soil water limitation to stomatal conductance is about 8.5 TgO3, while using a dynamic layer that ensures that plants maximize the water uptake from soil, we found a reduction of about 10 % in the amount of ozone removed by dry deposition ( ˜ 7.7 TgO3). Although dry deposition occurs from the top of canopy to ground level, it affects the concentration of gases remaining in the lower atmosphere, with a significant impact on ozone concentration (up to 4 ppb) extending from the surface to the upper troposphere (up to 650 hPa). Our results shed light on the importance of improving the parameterizations of processes occurring at plant level (i.e. from the soil to the canopy) as they have significant implications for concentration of gases in the lower troposphere and resulting risk assessments for vegetation or human health.

Stomatal conductance, mesophyll conductance, and transpiration efficiency in relation to leaf anatomy in rice and wheat genotypes under drought.

Science.gov (United States)

Ouyang, Wenjing; Struik, Paul C; Yin, Xinyou; Yang, Jianchang

2017-11-02

Increasing leaf transpiration efficiency (TE) may provide leads for growing rice like dryland cereals such as wheat (Triticum aestivum). To explore avenues for improving TE in rice, variations in stomatal conductance (gs) and mesophyll conductance (gm) and their anatomical determinants were evaluated in two cultivars from each of lowland, aerobic, and upland groups of Oryza sativa, one cultivar of O. glaberrima, and two cultivars of T. aestivum, under three water regimes. The TE of upland rice, O. glaberrima, and wheat was more responsive to the gm/gs ratio than that of lowland and aerobic rice. Overall, the explanatory power of the particular anatomical trait varied among species. Low stomatal density mostly explained the low gs in drought-tolerant rice, whereas rice genotypes with smaller stomata generally responded more strongly to drought. Compared with rice, wheat had a higher gm, which was associated with thicker mesophyll tissue, mesophyll and chloroplasts more exposed to intercellular spaces, and thinner cell walls. Upland rice, O. glaberrima, and wheat cultivars minimized the decrease in gm under drought by maintaining high ratios of chloroplasts to exposed mesophyll cell walls. Rice TE could be improved by increasing the gm/gs ratio via modifying anatomical traits. © The Author 2017. Published by Oxford University Press on behalf of the Society for Experimental Biology.

Effect of nitrogen supply on transpiration and stomatal behaviour of beans (Phaseolus vulgaris L. )

Energy Technology Data Exchange (ETDEWEB)

Shimshi, D

1970-01-01

The effect of nitrogen supply on the transpiration rate and stomatal opening of potted bean plants was studied in a series of experiments. The transpiration rates of N-supplied plants were higher than those of N-deficient plants when soil moisture was relatively high; as soil moisture approached the wilting range, the transpiration rates of N-supplied plants dropped to below those of N-deficient plants. In spite of the marked differences in transpiration rates, as influenced by soil moisture and nitrogen supply, the stomata appeared closed. By coating the upper or lower surfaces of the leaves with a vapor-impervious compound it was shown that stomatal apertures below the limit of microscopic resolution control the rate of transpiration. Under conditions that encourage stomatal opening (covering the plants with transparent plastic bags), the stomata of the N-deficient plants opened to a lesser degree than those of N-supplied plants. There was some evidence that when stomata were visibly open, transpiration rates were regulated by the degree of plant hydration rather than by the degree of stomatal opening. It is concluded that N-deficient plants fail to open their stomata as widely and to close them as tightly as N-supplied plants. 8 references, 2 tables.

Using Plant Temperature to Evaluate the Response of Stomatal Conductance to Soil Moisture Deficit

Directory of Open Access Journals (Sweden)

Ming-Han Yu

2015-10-01

Full Text Available Plant temperature is an indicator of stomatal conductance, which reflects soil moisture stresses. We explored the relationship between plant temperature and soil moisture to optimize irrigation schedules in a water-stress experiment using Firmiana platanifolia (L. f. Marsili in an incubator. Canopy temperature, leaf temperature, and stomatal conductance were measured using thermal imaging and a porometer. The results indicated that (1 stomatal conductance decreased with declines in soil moisture, and reflected average canopy temperature; (2 the variation of the leaf temperature distribution was a reliable indicator of soil moisture stress, and the temperature distribution in severely water-stressed leaves exhibited greater spatial variation than that in the presence of sufficient irrigation; (3 thermal indices (Ig and crop water stress index (CWSI were theoretically proportional to stomatal conductance (gs, Ig was certified to have linearity relationship with gs and CWSI have a logarithmic relationship with gs, and both of the two indices can be used to estimate soil moisture; and (4 thermal imaging data can reflect water status irrespective of long-term water scarcity or lack of sudden rainfall. This study applied thermal imaging methods to monitor plants and develop adaptable irrigation scheduling, which are important for the formulation of effective and economical agriculture and forestry policy.

Solar-induced chlorophyll fluorescence tracks the trend of canopy stomatal conductance and transpiration at diurnal and seasonal scales

Science.gov (United States)

Zhang, Y.; Shan, N.; Ju, W.; Chen, J.

2017-12-01

Transpiration is the process of plant water loss through the stomata on the leaf surface and plays a key role in the energy and water balance of the land surface. Plant stomata function as a control interface for regulating photosynthetic uptake of CO2 and transpiration, strongly linked to plant productivity. Stomatal conductance is fundamental to larger-scale regional prediction of carbon-water cycles and their feedbacks to climate. The widely used Ball-Berry model coupled photosynthesis to a semi-empirical model of stomatal conductance. However large uncertainties remain in simulation of carbon assimilation rate in ecosystem and regional scales. The strong correlations of solar-induced fluorescence (SIF) and GPP have been demonstrated and provides an important opportunity to accurately monitor photosynthetic activity and water exchange. In this presentation, we compared both canopy-observed SIF and satellite-derived SIF with tower-based canopy stomatal conductance from hourly to 8-day scales in forest and cropland ecosystem. Using the model of stomatal conductance based on SIF, the transpiration was estimated at hourly and daily scales and compared with flux tower measurements. The results showed that the seasonal pattern of canopy stomatal conductance agreed better with SIF compared to NDVI and their relationship was higher during sunny days for forest ecosystem. Canopy stomatal conductance correlated with both tower-observed SIF and SIF from the Global Ozone Monitoring Experiment-2. Estimation of transpiration from SIF performed well in both forest and cropland ecosystem. This remotely sensed approaches from SIF for modelling stomatal conductance opens a new era to analysis and simulation of coupled carbon and water cycles under climate change.

Ferns are less dependent on passive dilution by cell expansion to coordinate leaf vein and stomatal spacing than angiosperms.

Directory of Open Access Journals (Sweden)

Madeline R Carins Murphy

Full Text Available Producing leaves with closely spaced veins is a key innovation linked to high rates of photosynthesis in angiosperms. A close geometric link between veins and stomata in angiosperms ensures that investment in enhanced venous water transport provides the strongest net carbon return to the plant. This link is underpinned by "passive dilution" via expansion of surrounding cells. However, it is not known whether this 'passive dilution' mechanism is present in plant lineages other than angiosperms and is another key feature of the angiosperms' evolutionary success. Consequently, we sought to determine whether the 'passive dilution' mechanism is; (i exclusive to the angiosperms, (ii a conserved mechanism that evolved in the common ancestor of ferns and angiosperms, or (iii has evolved continuously over time. To do this we first we assessed the plasticity of vein and stomatal density and epidermal cell size in ferns in response to light environment. We then compared the relationships between these traits found among ferns with modelled relationships that assume vein and stomatal density respond passively to epidermal cell expansion, and with those previously observed in angiosperms. Vein density, stomatal density and epidermal cell size were linked in ferns with remarkably similar relationships to those observed in angiosperms, except that fern leaves had fewer veins per stomata. However, plasticity was limited in ferns and stomatal spacing was dependent on active stomatal differentiation as well as passive cell expansion. Thus, ferns (like angiosperms appear to coordinate vein and stomatal density with epidermal cell expansion to some extent to maintain a constant ratio between veins and stomata in the leaf. The different general relationships between vein density and stomatal density in ferns and angiosperms suggests the groups have different optimum balances between the production of vein tissue dedicated to water supply and stomatal tissue for gas

Experience with TL-102M for the treatment of radiation stomatitis

International Nuclear Information System (INIS)

Nishio, Juntaro; Matsuya, Tokuzo; Inoue, Kazuo; Miyazaki, Tadashi; Maeda, Noriaki.

1984-01-01

TL-102M was administered to 14 patients who had radiation stomatitis following radiation therapy for malignant tumors in the oral cavity. Regarding the degree of overall improvement, one of the 14 patients was evaluated as ''extremely improved'', eight as ''improved'', four as ''slightly improved'', and one as ''unchanged''. None of the patients had side effects. Adherent, powdered TL-102M was easy to take for patients. Most of the patients desired to continue to take this drug because of having neither painfulness nor adhesive feeling. The usage of TL-102M could be helpful in promoting the treatment for cancer, thus suggesting that it is useful in treating radiation stomatitis. (Namekawa, K.)

Phenomics allows identification of genomic regions affecting maize stomatal conductance with conditional effects of water deficit and evaporative demand.

Science.gov (United States)

Prado, Santiago Alvarez; Cabrera-Bosquet, Llorenç; Grau, Antonin; Coupel-Ledru, Aude; Millet, Emilie J; Welcker, Claude; Tardieu, François

2018-02-01

Stomatal conductance is central for the trades-off between hydraulics and photosynthesis. We aimed at deciphering its genetic control and that of its responses to evaporative demand and water deficit, a nearly impossible task with gas exchanges measurements. Whole-plant stomatal conductance was estimated via inversion of the Penman-Monteith equation from data of transpiration and plant architecture collected in a phenotyping platform. We have analysed jointly 4 experiments with contrasting environmental conditions imposed to a panel of 254 maize hybrids. Estimated whole-plant stomatal conductance closely correlated with gas-exchange measurements and biomass accumulation rate. Sixteen robust quantitative trait loci (QTLs) were identified by genome wide association studies and co-located with QTLs of transpiration and biomass. Light, vapour pressure deficit, or soil water potential largely accounted for the differences in allelic effects between experiments, thereby providing strong hypotheses for mechanisms of stomatal control and a way to select relevant candidate genes among the 1-19 genes harboured by QTLs. The combination of allelic effects, as affected by environmental conditions, accounted for the variability of stomatal conductance across a range of hybrids and environmental conditions. This approach may therefore contribute to genetic analysis and prediction of stomatal control in diverse environments. © 2017 John Wiley & Sons Ltd.

Drought responses of two gymnosperm species with contrasting stomatal regulation strategies under elevated [CO2] and temperature.

Science.gov (United States)

Duan, Honglang; O'Grady, Anthony P; Duursma, Remko A; Choat, Brendan; Huang, Guomin; Smith, Renee A; Jiang, Yanan; Tissue, David T

2015-07-01

Future climate regimes characterized by rising [CO2], rising temperatures and associated droughts may differentially affect tree growth and physiology. However, the interactive effects of these three factors are complex because elevated [CO2] and elevated temperature may generate differential physiological responses during drought. To date, the interactive effects of elevated [CO2] and elevated temperature on drought-induced tree mortality remain poorly understood in gymnosperm species that differ in stomatal regulation strategies. Water relations and carbon dynamics were examined in two species with contrasting stomatal regulation strategies: Pinus radiata D. Don (relatively isohydric gymnosperm; regulating stomata to maintain leaf water potential above critical thresholds) and Callitris rhomboidea R. Br (relatively anisohydric gymnosperm; allowing leaf water potential to decline as the soil dries), to assess response to drought as a function of [CO2] and temperature. Both species were grown in two [CO2] (C(a) (ambient, 400 μl l(-1)) and C(e) (elevated, 640 μl l(-1))) and two temperature (T(a) (ambient) and T(e) (ambient +4 °C)) treatments in a sun-lit glasshouse under well-watered conditions. Drought plants were then exposed to a progressive drought until mortality. Prior to mortality, extensive xylem cavitation occurred in both species, but significant depletion of non-structural carbohydrates was not observed in either species. Te resulted in faster mortality in P. radiata, but it did not modify the time-to-mortality in C. rhomboidea. C(e) did not delay the time-to-mortality in either species under drought or T(e) treatments. In summary, elevated temperature (+4 °C) had greater influence than elevated [CO2] (+240 μl l(-1)) on drought responses of the two studied gymnosperm species, while stomatal regulation strategies did not generally affect the relative contributions of hydraulic failure and carbohydrate depletion to mortality under severe drought. �

Analysis of Stomatal Patterning in Selected Mutants of MAPK Pathways

KAUST Repository

Felemban, Abrar

2016-01-01

-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

Stomatal acclimation to vapour pressure deficit doubles transpiration of small tree seedlings with warming

DEFF Research Database (Denmark)

Marchin, Renée M.; Broadhead, Alice A.; Bostic, Laura E.

2016-01-01

chamber VPD. Warming increased mean water use of Carya by 140% and Quercus by 150%, but had no significant effect on water use of Acer. Increased water use of ring-porous species was attributed to (1) higher air T and (2) stomatal acclimation to VPD resulting in higher gs and more sensitive stomata......Future climate change is expected to increase temperature (T) and atmospheric vapour pressure deficit (VPD) in many regions, but the effect of persistent warming on plant stomatal behaviour is highly uncertain. We investigated the effect of experimental warming of 1.9-5.1 °C and increased VPD of 0.......5-1.3 kPa on transpiration and stomatal conductance (gs ) of tree seedlings in the temperate forest understory (Duke Forest, North Carolina, USA). We observed peaked responses of transpiration to VPD in all seedlings, and the optimum VPD for transpiration (Dopt ) shifted proportionally with increasing...

Differential effects of elevated air humidity on stomatal closing ability of Kalanchoë blossfeldiana between the C

NARCIS (Netherlands)

Fanourakis, Dimitrios; Hyldgaard, Benita; Gebraegziabher, Habtamu; Bouranis, Dimitris; Körner, Oliver; Nielsen, Kai Lønne; Ottosen, Carl-Otto

2017-01-01

High relative air humidity (RH ≥ 85%) impairs stomatal functionality, attenuating plant capacity to cope with abiotic stress. Previous studies were limited to C₃ species, so the RH effect on stomatal physiology of CAM plants remains unexplored. We addressed the topic through

The effect of denture stability, occlusion, oral hygiene and smoking on denture-induced stomatitis

International Nuclear Information System (INIS)

Nimri, Gadeer Mukatash

2008-01-01

This longitudinal clinical investigation was undertaken to find out the effect of denture wearing habit (day versus day and night), denture hygiene and cigarette smoking habit on the frequency of denture induced stomatitis. Comparisons were made between 240 complete denture wearers, half of whom were asked to wear their dentures at the daytime only and the other half to wear the denture day and night. All these participants were male patients with a mean age of 57.6 years who had received maxillary complete acrylic dentures for the first time. Fifty percent of the samples were smokers. A standard method for examination of the mouth and denture construction, insertion and follow up were employed. Putative risk factors (denture wearing habits, denture hygiene and smoking) were investigated. Subjects were recalled 12 months after insertion to examine the quality of the denture and the condition of the maxillary mucosa. No significant correlation was found between deterioration of stability or occlusion and type of habitual use of the dentures (P > 0.05). Fourteen percent of the cases reported with inflamed maxillary mucosa. Deterioration of retention or occlusion separately showed no correlation with the condition of the mucosa. However, associated deterioration of both stability and occlusion proved to be significantly correlated with the occurrence of denture stomatitis (P < 0.05). Denture stomatitis was significantly more frequently with subjects wearing their dentures overnight compared with those who removed them (P < 0.05). A significant correlation was also found between cigarette smoking, poor oral hygiene and the presence of denture induced stomatitis (P < 0.05). Nocturnal denture wearing habit, deficient oral and denture hygiene, and cigarette smoking are all important predisposing factors to denture-induced stomatitis, however, none of these factors was the sole cause of mucosal inflammation. (author)

Application of thermography for monitoring stomatal conductance of Coffea arabica under different shading systems.

Science.gov (United States)

Craparo, A C W; Steppe, K; Van Asten, P J A; Läderach, P; Jassogne, L T P; Grab, S W

2017-12-31

Stomatal regulation is a key process in the physiology of Coffea arabica (C. arabica). Intrinsically linked to photosynthesis and water relations, it provides insights into the plant's adaptive capacity, survival and growth. The ability to rapidly quantify this parameter for C. arabica under different agroecological systems would be an indispensable tool. Using a Flir E6 MIR Camera, an index that is equivalent to stomatal conductance (I g ) was compared with stomatal conductance measurements (g s ) in a mature coffee plantation. In order to account for varying meteorological conditions between days, the methods were also compared under stable meteorological conditions in a laboratory and I g was also converted to absolute stomatal conductance values (g 1 ). In contrast to typical plant-thermography methods which measure indices once per day over an extended time period, we used high resolution hourly measurements over daily time series with 9 sun and 9 shade replicates. Eight daily time series showed a strong correlation between methods, while the remaining 10 were not significant. Including several other meteorological parameters in the calculation of g 1 did not contribute to any stronger correlation between methods. Total pooled data (combined daily series) resulted in a correlation of ρ=0.66 (P≤2.2e-16), indicating that our approach is particularly useful for situations where absolute values of stomatal conductance are not required, such as for comparative purposes, screening or trend analysis. We use the findings to advance the protocol for a more accurate methodology which may assist in quantifying advantageous microenvironment designs for coffee, considering the current and future climates of coffee growing regions. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

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

Indian Academy of Sciences (India)

Dr.YASODHA

seedling raising, field planting and maintenance of the mapping population. ... tereticornis and production of interspecific hybrids displaying hybrid vigour in terms of .... A total of 114, 115 and 129 SSR, ISSR and SRAP markers were generated .... stomatal traits with yield and adaptability would help to improve productivity of ...

Inhibitory effects of bee venom and its components against viruses in vitro and in vivo.

Science.gov (United States)

Uddin, Md Bashir; Lee, Byeong-Hoon; Nikapitiya, Chamilani; Kim, Jae-Hoon; Kim, Tae-Hwan; Lee, Hyun-Cheol; Kim, Choul Goo; Lee, Jong-Soo; Kim, Chul-Joong

2016-12-01

Bee venom (BV) from honey bee (Apis Melifera L.) contains at least 18 pharmacologically active components including melittin (MLT), phospholipase A 2 (PLA 2 ), and apamin etc. BV is safe for human treatments dose dependently and proven to possess different healing properties including antibacterial and antiparasitidal properties. Nevertheless, antiviral properties of BV have not well investigated. Hence, we identified the potential antiviral properties of BV and its component against a broad panel of viruses. Co-incubation of non-cytotoxic amounts of BV and MLT, the main component of BV, significantly inhibited the replication of enveloped viruses such as Influenza A virus (PR8), Vesicular Stomatitis Virus (VSV), Respiratory Syncytial Virus (RSV), and Herpes Simplex Virus (HSV). Additionally, BV and MLT also inhibited the replication of non-enveloped viruses such as Enterovirus-71 (EV-71) and Coxsackie Virus (H3). Such antiviral properties were mainly explained by virucidal mechanism. Moreover, MLT protected mice which were challenged with lethal doses of pathogenic influenza A H1N1 viruses. Therefore, these results provides the evidence that BV and MLT could be a potential source as a promising antiviral agent, especially to develop as a broad spectrum antiviral agent.

High Resistance of Human Parainfluenza Type 2 Virus Protein-Expressing Cells to the Antiviral and Anti-Cell Proliferative Activities of Alpha/Beta Interferons: Cysteine-Rich V-Specific Domain Is Required for High Resistance to the Interferons

OpenAIRE

Nishio, Machiko; Tsurudome, Masato; Ito, Morihiro; Kawano, Mitsuo; Komada, Hiroshi; Ito, Yasuhiko

2001-01-01

Human parainfluenza type 2 virus (hPIV-2)-infected HeLa (HeLa-CA) cells and hPIV-2 V-expressing HeLa (HeLa-V) cells show high resistance to alpha/beta interferons (IFN-α/β) irrespective of whether vesicular stomatitis virus or Sindbis virus is used as a challenge virus. When Sindbis virus is used, these cells show high susceptibility to human IFN-γ. Furthermore, the multiplication of HeLa-V cells is not inhibited by IFN-α/β. HeLa cells expressing the N-terminally truncated V protein show resi...

Characterization of uncultivable bat influenza virus using a replicative synthetic virus.

Directory of Open Access Journals (Sweden)

Bin Zhou

2014-10-01

Full Text Available Bats harbor many viruses, which are periodically transmitted to humans resulting in outbreaks of disease (e.g., Ebola, SARS-CoV. Recently, influenza virus-like sequences were identified in bats; however, the viruses could not be cultured. This discovery aroused great interest in understanding the evolutionary history and pandemic potential of bat-influenza. Using synthetic genomics, we were unable to rescue the wild type bat virus, but could rescue a modified bat-influenza virus that had the HA and NA coding regions replaced with those of A/PR/8/1934 (H1N1. This modified bat-influenza virus replicated efficiently in vitro and in mice, resulting in severe disease. Additional studies using a bat-influenza virus that had the HA and NA of A/swine/Texas/4199-2/1998 (H3N2 showed that the PR8 HA and NA contributed to the pathogenicity in mice. Unlike other influenza viruses, engineering truncations hypothesized to reduce interferon antagonism into the NS1 protein didn't attenuate bat-influenza. In contrast, substitution of a putative virulence mutation from the bat-influenza PB2 significantly attenuated the virus in mice and introduction of a putative virulence mutation increased its pathogenicity. Mini-genome replication studies and virus reassortment experiments demonstrated that bat-influenza has very limited genetic and protein compatibility with Type A or Type B influenza viruses, yet it readily reassorts with another divergent bat-influenza virus, suggesting that the bat-influenza lineage may represent a new Genus/Species within the Orthomyxoviridae family. Collectively, our data indicate that the bat-influenza viruses recently identified are authentic viruses that pose little, if any, pandemic threat to humans; however, they provide new insights into the evolution and basic biology of influenza viruses.

Characterization of uncultivable bat influenza virus using a replicative synthetic virus.

Science.gov (United States)

Zhou, Bin; Ma, Jingjiao; Liu, Qinfang; Bawa, Bhupinder; Wang, Wei; Shabman, Reed S; Duff, Michael; Lee, Jinhwa; Lang, Yuekun; Cao, Nan; Nagy, Abdou; Lin, Xudong; Stockwell, Timothy B; Richt, Juergen A; Wentworth, David E; Ma, Wenjun

2014-10-01

Bats harbor many viruses, which are periodically transmitted to humans resulting in outbreaks of disease (e.g., Ebola, SARS-CoV). Recently, influenza virus-like sequences were identified in bats; however, the viruses could not be cultured. This discovery aroused great interest in understanding the evolutionary history and pandemic potential of bat-influenza. Using synthetic genomics, we were unable to rescue the wild type bat virus, but could rescue a modified bat-influenza virus that had the HA and NA coding regions replaced with those of A/PR/8/1934 (H1N1). This modified bat-influenza virus replicated efficiently in vitro and in mice, resulting in severe disease. Additional studies using a bat-influenza virus that had the HA and NA of A/swine/Texas/4199-2/1998 (H3N2) showed that the PR8 HA and NA contributed to the pathogenicity in mice. Unlike other influenza viruses, engineering truncations hypothesized to reduce interferon antagonism into the NS1 protein didn't attenuate bat-influenza. In contrast, substitution of a putative virulence mutation from the bat-influenza PB2 significantly attenuated the virus in mice and introduction of a putative virulence mutation increased its pathogenicity. Mini-genome replication studies and virus reassortment experiments demonstrated that bat-influenza has very limited genetic and protein compatibility with Type A or Type B influenza viruses, yet it readily reassorts with another divergent bat-influenza virus, suggesting that the bat-influenza lineage may represent a new Genus/Species within the Orthomyxoviridae family. Collectively, our data indicate that the bat-influenza viruses recently identified are authentic viruses that pose little, if any, pandemic threat to humans; however, they provide new insights into the evolution and basic biology of influenza viruses.

Stomatal structure and physiology do not explain differences in water use among montane eucalypts.

Science.gov (United States)

Gharun, Mana; Turnbull, Tarryn L; Pfautsch, Sebastian; Adams, Mark A

2015-04-01

Understanding the regulation of water use at the whole-tree scale is critical to advancing the utility of physiological ecology, for example in its role in predictive hydrology of forested catchments. For three eucalypt species that dominate high-elevation catchments in south-eastern Australia, we examined if whole-tree water use could be related to three widely discussed regulators of water use: stomatal anatomy, sensitivity of stomata [i.e. stomatal conductance (g(s))] to environmental influences, and sapwood area. While daily tree water use varied sixfold among species, sap velocity and sapwood area varied in parallel. Combined, stomatal structure and physiology could not explain differences in species-specific water use. Species which exhibited the fastest (Eucalyptus delegatensis) and slowest (Eucalyptus pauciflora) rates of water use both exhibited greater capacity for physiological control of g(s) [indicated by sensitivity to vapour pressure deficit (VPD)] and a reduced capacity to limit g(s) anatomically [indicated by greater potential g(s) (g(max))]. Conversely, g(s) was insensitive to VPD and g(max) was lowest for Eucalyptus radiata, the species showing intermediate rates of water use. Improved knowledge of stomatal anatomy will help us to understand the capacity of species to regulate leaf-level water loss, but seems likely to remain of limited use for explaining rates of whole-tree water use in montane eucalypts at the catchment scale.

Intraspecific variation in stomatal traits, leaf traits and physiology reflects adaptation along aridity gradients in a South African shrub.

Science.gov (United States)

Carlson, Jane E; Adams, Christopher A; Holsinger, Kent E

2016-01-01

Trait-environment relationships are commonly interpreted as evidence for local adaptation in plants. However, even when selection analyses support this interpretation, the mechanisms underlying differential benefits are often unknown. This study addresses this gap in knowledge using the broadly distributed South African shrub Protea repens. Specifically, the study examines whether broad-scale patterns of trait variation are consistent with spatial differences in selection and ecophysiology in the wild. In a common garden study of plants sourced from 19 populations, associations were measured between five morphological traits and three axes describing source climates. Trait-trait and trait-environment associations were analysed in a multi-response model. Within two focal populations in the wild, selection and path analyses were used to test associations between traits, fecundity and physiological performance. Across 19 populations in a common garden, stomatal density increased with the source population's mean annual temperature and decreased with its average amount of rainfall in midsummer. Concordantly, selection analysis in two natural populations revealed positive selection on stomatal density at the hotter, drier site, while failing to detect selection at the cooler, moister site. Dry-site plants with high stomatal density also had higher stomatal conductances, cooler leaf temperatures and higher light-saturated photosynthetic rates than those with low stomatal density, but no such relationships were present among wet-site plants. Leaf area, stomatal pore index and specific leaf area in the garden also co-varied with climate, but within-population differences were not associated with fitness in either wild population. The parallel patterns of broad-scale variation, differences in selection and differences in trait-ecophysiology relationships suggest a mechanism for adaptive differentiation in stomatal density. Densely packed stomata may improve performance by

Opinion: the red-light response of stomatal movement is sensed by the redox state of the photosynthetic electron transport chain.

Science.gov (United States)

Busch, Florian A

2014-02-01

Guard cells regulate CO2 uptake and water loss of a leaf by controlling stomatal movement in response to environmental factors such as CO2, humidity, and light. The mechanisms by which stomata respond to red light are actively debated in the literature, and even after decades of research it is still controversial whether stomatal movement is related to photosynthesis or not. This review summarizes the current knowledge of the red-light response of stomata. A comparison of published evidence suggests that stomatal movement is controlled by the redox state of photosynthetic electron transport chain components, in particular the redox state of plastoquinone. Potential consequences for the modeling of stomatal conductance are discussed.

Ebolavirus Vaccines: Progress in the Fight Against Ebola Virus Disease.

Science.gov (United States)

Wu, Xiao-Xin; Yao, Hang-Ping; Wu, Nan-Ping; Gao, Hai-Nv; Wu, Hai-Bo; Jin, Chang-Zhong; Lu, Xiang-Yun; Xie, Tian-Shen; Li, Lan-Juan

2015-01-01

Ebolaviruses are highly infectious pathogens that cause lethal Ebola virus disease (EVD) in humans and non-human primates (NHPs). Due to their high pathogenicity and transmissibility, as well as the potential to be misused as a bioterrorism agent, ebolaviruses would threaten the health of global populations if not controlled. In this review, we describe the origin and structure of ebolaviruses and the development of vaccines from the beginning of the 1980s, including conventional ebolavirus vaccines, DNA vaccines, Ebola virus-like particles (VLPs), vaccinia virus-based vaccines, Venezuelan equine encephalitis virus (VEEV)-like replicon particles, Kunjin virus-based vaccine, recombinant Zaire Ebolavirusx2206;VP30, recombinant cytomegalovirus (CMV)-based vaccines, recombinant rabies virus (RABV)-based vaccines, recombinant paramyxovirus-based vaccines, adenovirus-based vaccines and vesicular stomatitis virus (VSV)-based vaccines. No licensed vaccine or specific treatment is currently available to counteract ebolavirus infection, although DNA plasmids and several viral vector approaches have been evaluated as promising vaccine platforms. These vaccine candidates have been confirmed to be successful in protecting NHPs against lethal infection. Moreover, these vaccine candidates were successfully advanced to clinical trials. The present review provides an update of the current research on Ebola vaccines, with the aim of providing an overview on current prospects in the fight against EVD. © 2015 The Author(s) Published by S. Karger AG, Basel.

Development and Characterization of A Multiplexed RT-PCR Species Specific Assay for Bovine and one for Porcine Foot-and-Mouth Disease Virus Rule-Out

Energy Technology Data Exchange (ETDEWEB)

Smith, S M; Danganan, L; Tammero, L; Vitalis, B; Lenhoff, R; Naraghi-arani, P; Hindson, B

2007-08-06

Lawrence Livermore National Laboratory (LLNL), in collaboration with the Department of Homeland Security (DHS) and the United States Department of Agriculture (USDA), Animal and Plant Health Inspection Services (APHIS) has developed candidate multiplexed assays that may potentially be used within the National Animal Health Laboratory Network (NAHLN), the National Veterinary Services Laboratory (Ames, Iowa) and the Plum Island Animal Disease Center (PIADC). This effort has the ability to improve our nation's capability to discriminate between foreign animal diseases and those that are endemic using a single assay, thereby increasing our ability to protect food and agricultural resources with a diagnostic test which could enhance the nation's capabilities for early detection of a foreign animal disease. In FY2005 with funding from the DHS, LLNL developed the first version (Version 1.0) of a multiplexed (MUX) nucleic-acid-based RT-PCR assay that included signatures for foot-and-mouth disease virus (FMDV) detection with rule-out tests for two other foreign animal diseases (FADs) of swine, Vesicular Exanthema of Swine (VESV) and Swine Vesicular Disease Virus (SVDV), and four other domestic viral diseases Bovine Viral Diarrhea Virus (BVDV), Bovine Herpes Virus 1 (BHV-1), Bluetongue virus (BTV) and Parapox virus complex (which includes Bovine Papular Stomatitis Virus [BPSV], Orf of sheep, and Pseudocowpox). In FY06, LLNL has developed Bovine and Porcine species-specific panel which included existing signatures from Version 1.0 panel as well as new signatures. The MUX RT-PCR porcine assay for detection of FMDV includes the FADs, VESV and SVD in addition to vesicular stomatitis virus (VSV) and porcine reproductive and respiratory syndrome (PRRS). LLNL has also developed a MUX RT-PCR bovine assay for detection of FMDV with rule out tests for the two bovine FADs malignant catarrhal fever (MCF), rinderpest virus (RPV) and the domestic diseases vesicular stomatitis

Clinical evaluation of the essential oil of "Satureja Hortensis" for the treatment of denture stomatitis

Directory of Open Access Journals (Sweden)

Ali Mohammad Sabzghabaee

2012-01-01

Full Text Available Background: The prevalence of denture stomatitis has been shown to vary from 15 to 65% in complete denture wearers. Satureja hortensis L. has been considered to have antinociceptive, anti-inflammatory, antifungal and antimicrobial activities in vitro and exhibits strong inhibitory effect on the growth of periodontal bacteria. The aim of this study was to evaluate the efficacy of a 1% gel formulation of S. hortensis essential oil for the treatment of denture stomatitis. Materials and Methods: A randomized, controlled clinical trial study was conducted on 80 patients (mean age 62.91±7.34 in two parallel groups treated either with S. hortensis essential oil 1% gel or placebo applied two times daily for two weeks. Denture stomatitis was diagnosed by clinical examination and paraclinical confirmation with sampling the palatal mucosa for Candida albicans. Data were analyzed using Chi-squared or Student′s t tests. Results: The erythematous lesions of palatal area were significantly reduced (P<0.0001 in the treatment group who applied 1% topical gel of S. hortensis essential oil and Candida colonies count were reduced significantly (P=0.001. Conclusion: Topical application of the essential oil of S. hortensis could be considered as an effective agent for the treatment of denture stomatitis.

Bovine lactoferrin and piroxicam as an adjunct treatment for lymphocytic-plasmacytic gingivitis stomatitis in cats.

Science.gov (United States)

Hung, Yi-Ping; Yang, Yi-Ping; Wang, Hsien-Chi; Liao, Jiunn-Wang; Hsu, Wei-Li; Chang, Chao-Chin; Chang, Shih-Chieh

2014-10-01

Feline lymphocytic-plasmacytic gingivitis/stomatitis (LPGS) or caudal stomatitis is an inflammatory disease that causes painfully erosive lesions and proliferations of the oral mucosa. The disease is difficult to cure and can affect cats at an early age, resulting in lifetime therapy. In this study, a new treatment using a combination of bovine lactoferrin (bLf) oral spray and oral piroxicam was investigated using a randomized double-blinded clinical trial in 13 cats with caudal stomatitis. Oral lesion grading and scoring of clinical signs were conducted during and after the trial to assess treatment outcome. Oral mucosal biopsies were used to evaluate histological changes during and after treatment. Clinical signs were significantly improved in 77% of the cats. In a 4-week study, clinical signs were considerably ameliorated by oral piroxicam during the first 2 weeks. In a 12-week study, the combined bLf oral spray and piroxicam, when compared with piroxicam alone, exhibited an enhanced effect that reduced the severity of the oral lesions (P = 0.059), while also significantly improving clinical signs (P piroxicam was safe and might be used to decrease the clinical signs of caudal stomatitis in cats. Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.

Compound stress response in stomatal closure: a mathematical model of ABA and ethylene interaction in guard cells

Directory of Open Access Journals (Sweden)

Beguerisse-Dıaz Mariano

2012-11-01

Full Text Available Abstract Background Stomata are tiny pores in plant leaves that regulate gas and water exchange between the plant and its environment. Abscisic acid and ethylene are two well-known elicitors of stomatal closure when acting independently. However, when stomata are presented with a combination of both signals, they fail to close. Results Toshed light on this unexplained behaviour, we have collected time course measurements of stomatal aperture and hydrogen peroxide production in Arabidopsis thaliana guard cells treated with abscisic acid, ethylene, and a combination of both. Our experiments show that stomatal closure is linked to sustained high levels of hydrogen peroxide in guard cells. When treated with a combined dose of abscisic acid and ethylene, guard cells exhibit increased antioxidant activity that reduces hydrogen peroxide levels and precludes closure. We construct a simplified model of stomatal closure derived from known biochemical pathways that captures the experimentally observed behaviour. Conclusions Our experiments and modelling results suggest a distinct role for two antioxidant mechanisms during stomatal closure: a slower, delayed response activated by a single stimulus (abscisic acid ‘or’ ethylene and another more rapid ‘and’ mechanism that is only activated when both stimuli are present. Our model indicates that the presence of this rapid ‘and’ mechanism in the antioxidant response is key to explain the lack of closure under a combined stimulus.

[Suitability of four stomatal conductance models in agro-pastoral ecotone in North China: A case study for potato and oil sunflower.

Science.gov (United States)

Huang, Ming Xia; Wang, Jing; Tang, Jian Zhao; Yu, Qiang; Zhang, Jun; Xue, Qing Yu; Chang, Qing; Tan, Mei Xiu

2016-11-18

The suitability of four popular empirical and semi-empirical stomatal conductance models (Jarvis model, Ball-Berry model, Leuning model and Medlyn model) was evaluated based on para-llel observation data of leaf stomatal conductance, leaf net photosynthetic rate and meteorological factors during the vigorous growing period of potato and oil sunflower at Wuchuan experimental station in agro-pastoral ecotone in North China. It was found that there was a significant linear relationship between leaf stomatal conductance and leaf net photosynthetic rate for potato, whereas the linear relationship appeared weaker for oil sunflower. The results of model evaluation showed that Ball-Berry model performed best in simulating leaf stomatal conductance of potato, followed by Leuning model and Medlyn model, while Jarvis model was the last in the performance rating. The root-mean-square error (RMSE) was 0.0331, 0.0371, 0.0456 and 0.0794 mol·m -2 ·s -1 , the normalized root-mean-square error (NRMSE) was 26.8%, 30.0%, 36.9% and 64.3%, and R-squared (R 2 ) was 0.96, 0.61, 0.91 and 0.88 between simulated and observed leaf stomatal conductance of potato for Ball-Berry model, Leuning model, Medlyn model and Jarvis model, respectively. For leaf stomatal conductance of oil sunflower, Jarvis model performed slightly better than Leuning model, Ball-Berry model and Medlyn model. RMSE was 0.2221, 0.2534, 0.2547 and 0.2758 mol·m -2 ·s -1 , NRMSE was 40.3%, 46.0%, 46.2% and 50.1%, and R 2 was 0.38, 0.22, 0.23 and 0.20 between simulated and observed leaf stomatal conductance of oil sunflower for Jarvis model, Leuning model, Ball-Berry model and Medlyn model, respectively. The path analysis was conducted to identify effects of specific meteorological factors on leaf stomatal conductance. The diurnal variation of leaf stomatal conductance was principally affected by vapour pressure saturation deficit for both potato and oil sunflower. The model evaluation suggested that the stomatal

Capability of the "Ball-Berry" model for predicting stomatal conductance and water use efficiency of potato leaves under different irrigation regimes

DEFF Research Database (Denmark)

Liu, Fulai; Andersen, Mathias N.; Jensen, Christian Richardt

2009-01-01

was used for model parameterization, where measurements of midday leaf gas exchange of potted potatoes were done during progressive soil drying for 2 weeks at tuber initiation and earlier bulking stages. The measured photosynthetic rate (An) was used as an input for the model. To account for the effects......The capability of the ‘Ball-Berry' model (BB-model) in predicting stomatal conductance (gs) and water use efficiency (WUE) of potato (Solanum tuberosum L.) leaves under different irrigation regimes was tested using data from two independent pot experiments in 2004 and 2007. Data obtained from 2004...... of soil water deficits on gs, a simple equation modifying the slope (m) based on the mean soil water potential (Ψs) in the soil columns was incorporated into the original BB-model. Compared with the original BB-model, the modified BB-model showed better predictability for both gs and WUE of potato leaves...

Inhibition by acrolein of light-induced stomatal opening through inhibition of inward-rectifying potassium channels in Arabidopsis thaliana.

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Islam, Md Moshiul; Ye, Wenxiu; Matsushima, Daiki; Khokon, Md Atiqur Rahman; Munemasa, Shintaro; Nakamura, Yoshimasa; Murata, Yoshiyuki

2015-01-01

Acrolein is a reactive α,β-unsaturated aldehyde derived from lipid peroxides, which are produced in plants under a variety of stress. We investigated effects of acrolein on light-induced stomatal opening using Arabidopsis thaliana. Acrolein inhibited light-induced stomatal opening in a dose-dependent manner. Acrolein at 100 μM inhibited plasma membrane inward-rectifying potassium (Kin) channels in guard cells. Acrolein at 100 μM inhibited Kin channel KAT1 expressed in a heterologous system using Xenopus leaves oocytes. These results suggest that acrolein inhibits light-induced stomatal opening through inhibition of Kin channels in guard cells.

Stomatal kinetics and photosynthetic gas exchange along a continuum of isohydric to anisohydric regulation of plant water status.

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Meinzer, Frederick C; Smith, Duncan D; Woodruff, David R; Marias, Danielle E; McCulloh, Katherine A; Howard, Ava R; Magedman, Alicia L

2017-08-01

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 continuum. Here, we evaluated kinetics of light-induced stomatal opening, activation of photosynthesis and features of quasi-steady-state photosynthetic gas exchange in 10 woody species selected to represent different degrees of anisohydry. Based on a previously developed proxy for the degree of anisohydry, species' leaf water potentials at turgor loss, we found consistent trends in photosynthetic gas exchange traits across a spectrum of isohydry to anisohydry. More anisohydric species had faster kinetics of stomatal opening and activation of photosynthesis, and these kinetics were closely coordinated within species. Quasi-steady-state stomatal conductance and measures of photosynthetic capacity and performance were also greater in more anisohydric species. Intrinsic water-use efficiency estimated from leaf gas exchange and stable carbon isotope ratios was lowest in the most anisohydric species. In comparisons between gas exchange traits, species rankings were highly consistent, leading to species-independent scaling relationships over the range of isohydry to anisohydry observed. © 2017 John Wiley & Sons Ltd.

The dilemma of saving water or being cool: What determines the stomatal response under a changing climate?

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Haghighi, Erfan; Kirchner, James W.; Entekhabi, Dara

2017-04-01

Stomata play a critical role in terrestrial water and carbon cycles, regulating the trade-off between photosynthetic carbon gain and water loss in leaves. They adjust their aperture in response to a number of physiological and environmental factors, yet the mechanisms driving this response, particularly under climate extremes, remain poorly understood. Partial or complete stomatal closure reduces plant water stress under water-limited or high atmospheric evaporative demand conditions, but at the cost of reduced productivity, elevated heat, leaf shedding, and mortality. A proper account of such complex stomatal behavior is of particular importance for current ecosystem models that poorly capture observed vegetation responses in the context of climate change which is predicted to cause more frequent and intense temperature extremes along with an increase in the frequency of drought in many regions in the future. This study seeks to explore stomatal responses to environmental change accounted for by a varying soil-plant resistance under different atmospheric and soil moisture conditions. To this end, we developed a physically based transpiration model that couples stomatal control of leaf gas exchange to the leaf surface energy balance and the entire plant hydraulic system by considering the interdependence of the guard cell water potential (or turgor pressure) and transpiration rates. Model simulations of diurnal variations in transpiration rates were in good agreement with field observations, and facilitated quantitative prediction of stomatal and xylem flow regulation under a wide range of environmental conditions. Preliminary results demonstrate how soil and plant hydraulic conductances regulating stomatal opening and closure can help mitigate climatic water deficit (e.g., at midday) by boosting evaporative cooling. Our results are expected to advance physical understanding of the water cycle in the soil-plant-atmosphere continuum, and shed light on observed

Stomatal clustering in Begonia associates with the kinetics of leaf gaseous exchange and influences water use efficiency.

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Papanatsiou, Maria; Amtmann, Anna; Blatt, Michael R

2017-04-01

Stomata are microscopic pores formed by specialized cells in the leaf epidermis and permit gaseous exchange between the interior of the leaf and the atmosphere. Stomata in most plants are separated by at least one epidermal pavement cell and, individually, overlay a single substomatal cavity within the leaf. This spacing is thought to enhance stomatal function. Yet, there are several genera naturally exhibiting stomata in clusters and therefore deviating from the one-cell spacing rule with multiple stomata overlaying a single substomatal cavity. We made use of two Begonia species to investigate whether clustering of stomata alters guard cell dynamics and gas exchange under different light and dark treatments. Begonia plebeja, which forms stomatal clusters, exhibited enhanced kinetics of stomatal conductance and CO2 assimilation upon light stimuli that in turn were translated into greater water use efficiency. Our findings emphasize the importance of spacing in stomatal clusters for gaseous exchange and plant performance under environmentally limited conditions. © The Author 2017. Published by Oxford University Press on behalf of the Society for Experimental Biology.

ABA-Mediated Stomatal Response in Regulating Water Use during the Development of Terminal Drought in Wheat

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Renu Saradadevi

2017-07-01

Full Text Available End-of-season drought or “terminal drought,” which occurs after flowering, is considered the most significant abiotic stress affecting crop yields. Wheat crop production in Mediterranean-type environments is often exposed to terminal drought due to decreasing rainfall and rapid increases in temperature and evapotranspiration during spring when wheat crops enter the reproductive stage. Under such conditions, every millimeter of extra soil water extracted by the roots benefits grain filling and yield and improves water use efficiency (WUE. When terminal drought develops, soil dries from the top, exposing the top part of the root system to dry soil while the bottom part is in contact with available soil water. Plant roots sense the drying soil and produce signals, which on transmission to shoots trigger stomatal closure to regulate crop water use through transpiration. However, transpiration is linked to crop growth and productivity and limiting transpiration may reduce potential yield. While an early and high degree of stomatal closure affects photosynthesis and hence biomass production, a late and low degree of stomatal closure exhausts available soil water rapidly which results in yield losses through a reduction in post-anthesis water use. The plant hormone abscisic acid (ABA is considered the major chemical signal involved in stomatal regulation. Wheat genotypes differ in their ability to produce ABA under drought and also in their stomatal sensitivity to ABA. In this viewpoint article we discuss the possibilities of exploiting genotypic differences in ABA response to soil drying in regulating the use of water under terminal drought. Root density distribution in the upper drying layers of the soil profile is identified as a candidate trait that can affect ABA accumulation and subsequent stomatal closure. We also examine whether leaf ABA can be designated as a surrogate characteristic for improved WUE in wheat to sustain grain yield under

Oral cryotherapy for the prevention of high-dose melphalan-induced stomatitis in allogeneic hematopoietic stem cell transplant recipients.

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Aisa, Yoshinobu; Mori, Takehiko; Kudo, Masumi; Yashima, Tomoko; Kondo, Sakiko; Yokoyama, Akihiro; Ikeda, Yasuo; Okamoto, Shinichiro

2005-04-01

The purpose of this study was to evaluate the efficacy of oral cryotherapy to prevent high-dose melphalan-induced stomatitis. Eighteen consecutive recipients of allogeneic hematopoietic stem cell transplant conditioned with high-dose melphalan (140 mg/m2) in combination with fludarabine alone or with fludarabine and additional chemotherapy or radiation were enrolled. The severity of stomatitis was graded according to the National Cancer Institute Common Toxicity Criteria. Patients were kept on oral cryotherapy using ice chips and ice-cold water shortly before, during, and for additional 90 min after completion of melphalan administration. Only two of 18 patients (11.1%) developed grade 2 or 3 stomatitis while six of seven patients in the historical control developed it (85.7%; P=0.001). These results suggested that oral cryotherapy could effectively prevent stomatitis caused by high-dose melphalan, and we recommend that it should be incorporated into the conditioning regimen with high-dose melphalan.

Virus detection by PCR following vaccination of naive calves with intranasal or injectable multivalent modified-live viral vaccines.

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Walz, Paul H; Newcomer, Benjamin W; Riddell, Kay P; Scruggs, Daniel W; Cortese, Victor S

2017-09-01

We evaluated duration of PCR-positive results following administration of modified-live viral (MLV) vaccines to beef calves. Twenty beef calves were randomly assigned to either group 1 and vaccinated intranasally with a MLV vaccine containing bovine alphaherpesvirus 1 (BoHV-1), bovine respiratory syncytial virus (BRSV), and bovine parainfluenza virus 3 (BPIV-3), or to group 2 and vaccinated subcutaneously with a MLV vaccine containing bovine viral diarrhea virus 1 and 2 (BVDV-1, -2), BoHV-1, BRSV, and BPIV-3. Deep nasopharyngeal swabs (NPS) and transtracheal washes (TTW) were collected from all calves, and whole blood was collected from group 2 calves and tested by PCR. In group 1, the proportions of calves that tested PCR-positive to BVDV, BoHV-1, BRSV, and BPIV-3 on any sample at any time were 0%, 100%, 100%, and 10%, respectively. In group 1 calves, 100% of calves became PCR-positive for BoHV-1 by day 3 post-vaccination and 100% of calves became PCR-positive for BRSV by day 7 post-vaccination. In group 2, the proportions of calves that tested positive to BVDV, BoHV-1, BRSV, and BPIV-3 on any sample at any time were 50%, 40%, 10%, and 0%, respectively. All threshold cycle (Ct) values were >30 in group 2 calves, irrespective of virus; however, Ct values PCR-positive results for BoHV-1 and BRSV. All calves were PCR-negative for all viruses after day 28. Following intranasal MLV viral vaccination, PCR results and Ct values for BRSV and BoHV-1 suggest that attempts to differentiate vaccine virus from natural infection is unreliable.

A Two-Big-Leaf Model for Canopy Temperature, Photosynthesis, and Stomatal Conductance.

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Dai, Yongjiu; Dickinson, Robert E.; Wang, Ying-Ping

2004-06-01

The energy exchange, evapotranspiration, and carbon exchange by plant canopies depend on leaf stomatal control. The treatment of this control has been required by land components of climate and carbon models. Physiological models can be used to simulate the responses of stomatal conductance to changes in atmospheric and soil environments. Big-leaf models that treat a canopy as a single leaf tend to overestimate fluxes of CO2 and water vapor. Models that differentiate between sunlit and shaded leaves largely overcome these problems.A one-layered, two-big-leaf submodel for photosynthesis, stomatal conductance, leaf temperature, and energy fluxes is presented in this paper. It includes 1) an improved two stream approximation model of radiation transfer of the canopy, with attention to singularities in its solution and with separate integrations of radiation absorption by sunlit and shaded fractions of canopy; 2) a photosynthesis stomatal conductance model for sunlit and shaded leaves separately, and for the simultaneous transfers of CO2 and water vapor into and out of the leaf—leaf physiological properties (i.e., leaf nitrogen concentration, maximum potential electron transport rate, and hence photosynthetic capacity) vary throughout the plant canopy in response to the radiation weight time-mean profile of photosynthetically active radiation (PAR), and the soil water limitation is applied to both maximum rates of leaf carbon uptake by Rubisco and electron transport, and the model scales up from leaf to canopy separately for all sunlit and shaded leaves; 3) a well-built quasi-Newton Raphson method for simultaneous solution of temperatures of the sunlit and shaded leaves.The model was incorporated into the Common Land Model (CLM) and is denoted CLM 2L. It was driven with observational atmospheric forcing from two forest sites [Anglo-Brazilian Amazonian Climate Observation Study (ABRACOS) and Boreal Ecosystem Atmosphere Study (BOREAS)] for 2 yr of simulation. The

A fast method to detect the occurrence of nonhomogeneous distribution of stomatal aperture in heterobaric plant leaves : Experiments with Arbutus unedo L. during the diurnal course.

Science.gov (United States)

Beyschlag, W; Pfanz, H

1990-01-01

Pressure infiltration of water into a leaf via the stomatal pores can be used to quickly determine whether all stomata are open, or as recently described for several mesophytic and xerophytic species, whether there is a non-homogeneous distribution of stomatal opening (stomatal patchiness) on the leaf surface. Information about this phenomenon is important since the commonly used algorithms for calculation of leaf conductance from water vapor exchange measurements imply homogeneously open stomata, which in the occurrence of stomatal patchiness will lead to erroneous results. Infiltration experiments in a growth chamber with leaves of the Mediterranean evergreen shrub Arbutus unedo, carried out under simulated Mediterranean summer day conditions, where the species typically exhibits a strong midday stomatal closure, revealed a temporary occurrence of stomatal patchiness during the phase of stomatal closure in the late morning and during the stomatal reopening in the afternoon. Leaves were, however, found to be fully (i.e. homogeneously) infiltratable in the morning and in the evening. At midday during maximum stomatal closure, leaves were almost non-infiltratable. During the day, the infiltrated amount of water was found to be linearly correlated with porometer measurements of leaf conductance of the same leaves, carried out with the attached leaves immediately before infiltration.

Applicability of common stomatal conductance models in maize under varying soil moisture conditions.

Science.gov (United States)

Wang, Qiuling; He, Qijin; Zhou, Guangsheng

2018-07-01

In the context of climate warming, the varying soil moisture caused by precipitation pattern change will affect the applicability of stomatal conductance models, thereby affecting the simulation accuracy of carbon-nitrogen-water cycles in ecosystems. We studied the applicability of four common stomatal conductance models including Jarvis, Ball-Woodrow-Berry (BWB), Ball-Berry-Leuning (BBL) and unified stomatal optimization (USO) models based on summer maize leaf gas exchange data from a soil moisture consecutive decrease manipulation experiment. The results showed that the USO model performed best, followed by the BBL model, BWB model, and the Jarvis model performed worst under varying soil moisture conditions. The effects of soil moisture made a difference in the relative performance among the models. By introducing a water response function, the performance of the Jarvis, BWB, and USO models improved, which decreased the normalized root mean square error (NRMSE) by 15.7%, 16.6% and 3.9%, respectively; however, the performance of the BBL model was negative, which increased the NRMSE by 5.3%. It was observed that the models of Jarvis, BWB, BBL and USO were applicable within different ranges of soil relative water content (i.e., 55%-65%, 56%-67%, 37%-79% and 37%-95%, respectively) based on the 95% confidence limits. Moreover, introducing a water response function, the applicability of the Jarvis and BWB models improved. The USO model performed best with or without introducing the water response function and was applicable under varying soil moisture conditions. Our results provide a basis for selecting appropriate stomatal conductance models under drought conditions. Copyright © 2018 Elsevier B.V. All rights reserved.

Protective immunity and safety of a genetically modified influenza virus vaccine.

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Rafael Polidoro Alves Barbosa

Full Text Available Recombinant influenza viruses are promising viral platforms to be used as antigen delivery vectors. To this aim, one of the most promising approaches consists of generating recombinant viruses harboring partially truncated neuraminidase (NA segments. To date, all studies have pointed to safety and usefulness of this viral platform. However, some aspects of the inflammatory and immune responses triggered by those recombinant viruses and their safety to immunocompromised hosts remained to be elucidated. In the present study, we generated a recombinant influenza virus harboring a truncated NA segment (vNA-Δ and evaluated the innate and inflammatory responses and the safety of this recombinant virus in wild type or knock-out (KO mice with impaired innate (Myd88 -/- or acquired (RAG -/- immune responses. Infection using truncated neuraminidase influenza virus was harmless regarding lung and systemic inflammatory response in wild type mice and was highly attenuated in KO mice. We also demonstrated that vNA-Δ infection does not induce unbalanced cytokine production that strongly contributes to lung damage in infected mice. In addition, the recombinant influenza virus was able to trigger both local and systemic virus-specific humoral and CD8+ T cellular immune responses which protected immunized mice against the challenge with a lethal dose of homologous A/PR8/34 influenza virus. Taken together, our findings suggest and reinforce the safety of using NA deleted influenza viruses as antigen delivery vectors against human or veterinary pathogens.

Inactivation of RNA Viruses by Gamma Irradiation: A Study on Mitigating Factors

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Adam J. Hume

2016-07-01

Full Text Available Effective inactivation of biosafety level 4 (BSL-4 pathogens is vital in order to study these agents safely. Gamma irradiation is a commonly used method for the inactivation of BSL-4 viruses, which among other advantages, facilitates the study of inactivated yet morphologically intact virions. The reported values for susceptibility of viruses to inactivation by gamma irradiation are sometimes inconsistent, likely due to differences in experimental protocols. We analyzed the effects of common sample attributes on the inactivation of a recombinant vesicular stomatitis virus expressing the Zaire ebolavirus glycoprotein and green fluorescent protein. Using this surrogate virus, we found that sample volume and protein content of the sample modulated viral inactivation by gamma irradiation but that air volume within the sample container and the addition of external disinfectant surrounding the sample did not. These data identify several factors which alter viral susceptibility to inactivation and highlight the usefulness of lower biosafety level surrogate viruses for such studies. Our results underscore the need to validate inactivation protocols of BSL-4 pathogens using “worst-case scenario” procedures to ensure complete sample inactivation.

Study on preventive effects of i.v. administration of flavin adenine dinucleotide (FAD) before irradiation on radiation stomatitis

International Nuclear Information System (INIS)

Nagai, Masao; Houzawa, Jiro; Hakamada, Masaru

1984-01-01

In order to compare the preventive effect on radiation stomatitis, flavin adenine dinucleotide (FAD) or vitamin C was administered intravenously until the blood level reached the maximum at the time of irradiation. Thirtyfive patients with cranial or cervical tumors were allocated into the group with FAD (15), the group with vitamin C (10), and the group with irradiation alone (10). The incidence of stomititis was significantly lower and the number of patients in whom the drug was withdrawn due to stomatitis was extremely smaller in the group with FAD than in the other groups. FAD administered before irradiation was considered very useful in preventing radiation stomatitis. (Namekawa, K.)

Plester sariawan efektif dalam mempercepat penyembuhan stomatitis aftosa rekuren dan ulkus traumatikus

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Rahmi Amtha

2017-12-01

Full Text Available Mouth ulcer plaster is effective in accelerating the healing of recurrent aphthous stomatitis and traumatic ulcers. Recurrent aphthous stomatitis (RAS is one of the most commonly occurring oral diseases. The prevalence of oral ulceration worldwide is 4%, with RAS having the largest proportion (25%. Recurrent aphthous stomatitis is oral ulceration which has a self-limiting disease, but the specific medication to reduce pain caused by lesion is still less varied nowadays. This study aimed to examine the differences in the effectiveness between topical application of hyaluronic acid (HA, mouth ulcer plaster (MUP and 0.1% triamcinolone acetonide (TA as a positive control in the healing of RAS and traumatic ulcers (TU. This was a quasi-experimental study by measuring the lesion diameter as well as visual analogue scale (VAS pre- and post-administration of three types of medication. Kruskal-walis test results show that there are differences in effectiveness (p=0.000 of the three types of medication to cure RAS and TU. There are signicant differences in the reduction of RAS and TU lesion diameter (p = 0.015 and VAS (p = 0.038 with the use of HA and MUP on the 4th day. There is no signicant difference in effectiveness (diameter and VAS of MUP and TA medication on the fourth day (p = 0.880 and p = 1.000 respectively. There is no signicant difference among HA, MUP and TA on the healing of the lesions on the seventh day (p>0.05. It can be concluded that the effectiveness of MUP is similar to that of topical medications containing corticosteroids in the healing of RAS and traumatic ulcers. ABSTRAK Stomatitis aftosa rekuren (SAR merupakan salah satu penyakit mulut yang paling umum terjadi. Prevalensi ulserasi mulut di seluruh dunia adalah 4%, dengan SAR menempati urutan terbesar yaitu 25%. Stomatitis aftosa rekuren merupakan ulserasi mulut yang memiliki self-limiting disease, namun sediaan obat yang spesifik untuk mengurangi rasa sakit yang

Chlorella induces stomatal closure via NADPH oxidase-dependent ROS production and its effects on instantaneous water use efficiency in Vicia faba.

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Yan Li

Full Text Available Reactive oxygen species (ROS have been established to participate in stomatal closure induced by live microbes and microbe-associated molecular patterns (MAMPs. Chlorella as a beneficial microorganism can be expected to trigger stomatal closure via ROS production. Here, we reported that Chlorella induced stomatal closure in a dose-and time-dependent manner in epidermal peels of Vicia faba. Using pharmacological methods in this work, we found that the Chlorella-induced stomatal closure was almost completely abolished by a hydrogen peroxide (H2O2 scavenger, catalase (CAT, significantly suppressed by an NADPH oxidase inhibitor, diphenylene iodonium chloride (DPI, and slightly affected by a peroxidase inhibitor, salicylhydroxamic acid (SHAM, suggesting that ROS production involved in Chlorella-induced stomatal closure is mainly mediated by DPI-sensitive NADPH oxidase. Additionally, Exogenous application of optimal concentrations of Chlorella suspension improved instantaneous water use efficiency (WUEi in Vicia faba via a reduction in leaf transpiration rate (E without a parallel reduction in net photosynthetic rate (Pn assessed by gas-exchange measurements. The chlorophyll fluorescence and content analysis further demonstrated that short-term use of Chlorella did not influence plant photosynthetic reactions center. These results preliminarily reveal that Chlorella can trigger stomatal closure via NADPH oxidase-dependent ROS production in epidermal strips and improve WUEi in leave levels.

Host DNA synthesis-suppressing factor in culture fluid of tissue cultures infected with measles virus

International Nuclear Information System (INIS)

Minagawa, T.; Nakaya, C.; Iida, H.

1974-01-01

Host DNA synthesis is suppressed by the culture fluid of cell cultures infected with measles virus. This activity in the culture fluid is initiated somewhat later than the growth of infectious virus. Ninety percent of host DNA synthesis in HeLa cells is inhibited by culture fluid of 3-day-old cell cultures of Vero or HeLa cells infected with measles virus. This suppressing activity is not a property of the virion, but is due to nonvirion-associated componentnent which shows none of the activities of measles virus such as hemagglutination, hemolysis, or cell fusion nor does it have the antigenicity of measles virus as tested by complement-fixation or hemagglutination-inhibiting antibody blocking tests. Neutralization of the activity of this component is not attained with the pooled sera of convalescent measles patients. This component has molecular weights of about 45,000, 20,000, and 3,000 and appears to be a heat-stable protein. The production of host DNA suppressing factor (DSF) is blocked by cycloheximide. Neither uv-inactivated nor antiserum-neutralized measles virus produce DSF. Furthermore, such activity of nonvirion-associated component is not detected in the culture fluid of cultures infected with other RNA viruses such as poliovirus, vesicular stomatitis virus, or Sindbis virus. (auth)

Stomatal responses to CO2 during a diel Crassulacean acid metabolism cycle in Kalanchoe daigremontiana and Kalanchoe pinnata.

Science.gov (United States)

von Caemmerer, Susanne; Griffiths, Howard

2009-05-01

To investigate the diurnal variation of stomatal sensitivity to CO2, stomatal response to a 30 min pulse of low CO2 was measured four times during a 24 h time-course in two Crassulacean acid metabolism (CAM) species Kalanchoe daigremontiana and Kalanchoe pinnata, which vary in the degree of succulence, and hence, expression and commitment to CAM. In both species, stomata opened in response to a reduction in pCO2 in the dark and in the latter half of the light period, and thus in CAM species, chloroplast photosynthesis is not required for the stomatal response to low pCO2. Stomata did not respond to a decreased pCO2 in K. daigremontiana in the light when stomata were closed, even when the supply of internal CO2 was experimentally reduced. We conclude that stomatal closure during phase III is not solely mediated by high internal pCO2, and suggest that in CAM species the diurnal variability in the responsiveness of stomata to pCO2 could be explained by hypothesizing the existence of a single CO2 sensor which interacts with other signalling pathways. When not perturbed by low pCO2, CO2 assimilation rate and stomatal conductance were correlated both in the light and in the dark in both species.

Characterization of Dengue Virus Resistance to Brequinar in Cell Culture▿

Science.gov (United States)

Qing, Min; Zou, Gang; Wang, Qing-Yin; Xu, Hao Ying; Dong, Hongping; Yuan, Zhiming; Shi, Pei-Yong

2010-01-01

Brequinar is an inhibitor of dihydroorotate dehydrogenase, an enzyme that is required for de novo pyrimidine biosynthesis. Here we report that brequinar has activity against a broad spectrum of viruses. The compound not only inhibits flaviviruses (dengue virus, West Nile virus, yellow fever virus, and Powassan virus) but also suppresses a plus-strand RNA alphavirus (Western equine encephalitis virus) and a negative-strand RNA rhabdovirus (vesicular stomatitis virus). Using dengue virus serotype 2 (DENV-2) as a model, we found that brequinar suppressed the viral infection cycle mainly at the step of RNA synthesis. Supplementing the culture medium with pyrimidines (cytidine or uridine) but not purines (adenine or guanine) could be used to reverse the inhibitory effect of the compound. Continuous culturing of DENV-2 in the presence of brequinar generated viruses that were partially resistant to the inhibitor. Sequencing of the resistant viruses revealed two amino acid mutations: one mutation (M260V) located at a helix in the domain II of the viral envelope protein and another mutation (E802Q) located at the priming loop of the nonstructural protein 5 (NS5) polymerase domain. Functional analysis of the mutations suggests that the NS5 mutation exerts resistance through enhancement of polymerase activity. The envelope protein mutation reduced the efficiency of virion assembly/release; however, the mutant virus became less sensitive to brequinar inhibition at the step of virion assembly/release. Taken together, the results indicate that (i) brequinar blocks DENV RNA synthesis through depletion of intracellular pyrimidine pools and (ii) the compound may also exert its antiviral activity through inhibition of virion assembly/release. PMID:20606073

Differences in the response sensitivity of stomatal index to atmospheric CO2 among four genera of Cupressaceae conifers.

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Haworth, Matthew; Heath, James; McElwain, Jennifer C

2010-03-01

The inverse relationship between stomatal density (SD: number of stomata per mm(2) leaf area) and atmospheric concentration of CO2 ([CO2]) permits the use of plants as proxies of palaeo-atmospheric CO2. Many stomatal reconstructions of palaeo-[CO2] are based upon multiple fossil species. However, it is unclear how plants respond to [CO2] across genus, family or ecotype in terms of SD or stomatal index (SI: ratio of stomata to epidermal cells). This study analysed the stomatal numbers of conifers from the ancient family Cupressaceae, in order to examine the nature of the SI-[CO2] relationship, and potential implications for stomatal reconstructions of palaeo-[CO2]. Methods Stomatal frequency measurements were taken from historical herbarium specimens of Athrotaxis cupressoides, Tetraclinis articulata and four Callitris species, and live A. cupressoides grown under CO2-enrichment (370, 470, 570 and 670 p.p.m. CO2). T. articulata, C. columnaris and C. rhomboidea displayed significant reductions in SI with rising [CO2]; by contrast, A. cupressoides, C. preissii and C. oblonga show no response in SI. However, A. cupressoides does reduce SI to increases in [CO2] above current ambient (approx. 380 p.p.m. CO2). This dataset suggests that a shared consistent SI-[CO2] relationship is not apparent across the genus Callitris. Conclusions The present findings suggest that it is not possible to generalize how conifer species respond to fluctuations in [CO2] based upon taxonomic relatedness or habitat. This apparent lack of a consistent response, in conjunction with high variability in SI, indicates that reconstructions of absolute palaeo-[CO2] based at the genus level, or upon multiple species for discrete intervals of time are not as reliable as those based on a single or multiple temporally overlapping species.

Relationship between ammonia stomatal compensation point and nitrogen metabolism in arable crops: Current status of knowledge and potential modelling approaches

International Nuclear Information System (INIS)

Massad, Raia Silvia; Loubet, Benjamin; Tuzet, Andree; Cellier, Pierre

2008-01-01

The ammonia stomatal compensation point of plants is determined by leaf temperature, ammonium concentration ([NH 4 + ] apo ) and pH of the apoplastic solution. The later two depend on the adjacent cells metabolism and on leaf inputs and outputs through the xylem and phloem. Until now only empirical models have been designed to model the ammonia stomatal compensation point, except the model of Riedo et al. (2002. Coupling soil-plant-atmosphere exchange of ammonia with ecosystem functioning in grasslands. Ecological Modelling 158, 83-110), which represents the exchanges between the plant's nitrogen pools. The first step to model the ammonia stomatal compensation point is to adequately model [NH 4 + ] apo . This [NH 4 + ] apo has been studied experimentally, but there are currently no process-based quantitative models describing its relation to plant metabolism and environmental conditions. This study summarizes the processes involved in determining the ammonia stomatal compensation point at the leaf scale and qualitatively evaluates the ability of existing whole plant N and C models to include a model for [NH 4 + ] apo . - A model for ammonia stomatal compensation point at the leaf level scale was developed

Secreted herpes simplex virus-2 glycoprotein G modifies NGF-TrkA signaling to attract free nerve endings to the site of infection.

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Jorge Rubén Cabrera

2015-01-01

Full Text Available Herpes simplex virus type 1 (HSV-1 and HSV-2 are highly prevalent viruses that cause a variety of diseases, from cold sores to encephalitis. Both viruses establish latency in peripheral neurons but the molecular mechanisms facilitating the infection of neurons are not fully understood. Using surface plasmon resonance and crosslinking assays, we show that glycoprotein G (gG from HSV-2, known to modulate immune mediators (chemokines, also interacts with neurotrophic factors, with high affinity. In our experimental model, HSV-2 secreted gG (SgG2 increases nerve growth factor (NGF-dependent axonal growth of sympathetic neurons ex vivo, and modifies tropomyosin related kinase (TrkA-mediated signaling. SgG2 alters TrkA recruitment to lipid rafts and decreases TrkA internalization. We could show, with microfluidic devices, that SgG2 reduced NGF-induced TrkA retrograde transport. In vivo, both HSV-2 infection and SgG2 expression in mouse hindpaw epidermis enhance axonal growth modifying the termination zone of the NGF-dependent peptidergic free nerve endings. This constitutes, to our knowledge, the discovery of the first viral protein that modulates neurotrophins, an activity that may facilitate HSV-2 infection of neurons. This dual function of the chemokine-binding protein SgG2 uncovers a novel strategy developed by HSV-2 to modulate factors from both the immune and nervous systems.

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...

Variation in RNA virus mutation rates across host cells.

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Marine Combe

2014-01-01

Full Text Available It is well established that RNA viruses exhibit higher rates of spontaneous mutation than DNA viruses and microorganisms. However, their mutation rates vary amply, from 10(-6 to 10(-4 substitutions per nucleotide per round of copying (s/n/r and the causes of this variability remain poorly understood. In addition to differences in intrinsic fidelity or error correction capability, viral mutation rates may be dependent on host factors. Here, we assessed the effect of the cellular environment on the rate of spontaneous mutation of the vesicular stomatitis virus (VSV, which has a broad host range and cell tropism. Luria-Delbrück fluctuation tests and sequencing showed that VSV mutated similarly in baby hamster kidney, murine embryonic fibroblasts, colon cancer, and neuroblastoma cells (approx. 10(-5 s/n/r. Cell immortalization through p53 inactivation and oxygen levels (1-21% did not have a significant impact on viral replication fidelity. This shows that previously published mutation rates can be considered reliable despite being based on a narrow and artificial set of laboratory conditions. Interestingly, we also found that VSV mutated approximately four times more slowly in various insect cells compared with mammalian cells. This may contribute to explaining the relatively slow evolution of VSV and other arthropod-borne viruses in nature.

Hendra and Nipah viruses: pathogenesis, animal models and recent breakthroughs in vaccination

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Weingartl HM

2015-09-01

Full Text Available Hana M Weingartl National Centre for Foreign Animal Disease, Canadian Food Inspection Agency, Winnipeg, MB, Canada Abstract: Hendra and Nipah viruses are two highly pathogenic zoonotic members of the genus Henipavirus, family Paramyxoviridae, requiring work under biosafety level 4 conditions due to a lack of effective therapy and human vaccines. Several vaccine candidates were protective in animal models: recombinant vaccinia virus expressing Nipah virus (NiV F and G proteins in hamsters against NiV; recombinant ALVAC–NiV F and G in swine against NiV; recombinant Hendra virus (HeV soluble G protein (sGHeV against HeV and NiV in cats, ferrets, horses, and African green monkeys (AGM; recombinant vesicular stomatitis virus-based vectors expressing NiV F or G against NiV in hamsters and ferrets; measles virus-based NiV G vaccine candidate in hamsters and AGMs against NiV; and adenoassociated virus expressing NiG protein, which protected hamsters against NiV. The sGHeV was licensed for use in horses (Equivac HeV® in 2012. It is the first vaccine candidate licensed against a biosafety level 4 agent. With the development of suitable animal models (ferret, hamster and, importantly, AGM, progress can be made toward development of a human vaccine.Keywords: henipavirus, equine, swine, human infection, animal models, vaccine candidates

Stomatal distribution, stomatal density and daily leaf movement in Acacia aroma (Leguminosae Distribución y densidad estomática y movimiento diario de la hoja en Acacia aroma (Leguminosae

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Marcelo P. Hernández

2010-12-01

Full Text Available Acacia aroma Gillies ex Hook. & Arn. grows in the Chacoan and Yungas Biogeographic Provinces, Argentina. It has numerous medicinal applications, sweet and edible fruits, and it may be used as forage. The objective of the present contribution was to analyse the stomatal distribution and stomatal density on the secondary leaflet surfaces, in different parts of the leaf, and at different tree crown levels, establishing the leaf movement and environmental condition relationships. The work was performed with fresh material and herbarium specimens, using conventional anatomical techniques. Stomatal distribution on the secondary leaflet surfaces was established, and differences in stomatal density among basal, medium and apical leaflets were found. A decrease in stomatal density from the lower level to the upper level of the tree crown would be connected with that. The stomatal distribution and density appear related to the secondary leaflet shape and its position on the secondary rachis, interacting with the daily secondary leaflets and leaf movement, and the weather conditions. It is interesting that the medium value of stomata density were found in the middle part of the leaf and at the middle level of the tree crown. Original illustrations are given.Acacia aroma crece en las Provincias Biogeográficas Chaqueña y de las Yungas, Argentina. Este árbol posee numerosas aplicaciones en medicina popular, sus frutos son comestibles y puede ser usada como forraje. Los objetivos de la presente contribución fueron: establecer la distribución y densidad de los estomas en el folíolo secundario, en distintos folíolos secundarios de la misma hoja y en los folíolos secundarios de las hojas de la parte basal, media y superior de la copa del árbol, estableciendo relaciones con el movimiento diario de las hojas y condiciones ambientales. Para el estudio se utilizó material fresco y ejemplares de herbario empleando técnicas de anatomía convencionales. Se

Effect of Light Quality on Stomatal Opening in Leaves of Xanthium strumarium L.

Science.gov (United States)

Sharkey, T D; Raschke, K

1981-11-01

Flux response curves were determined at 16 wavelengths of light for the conductance for water vapor of the lower epidermis of detached leaves of Xanthium strumarium L. An action spectrum of stomatal opening resulted in which blue light (wavelengths between 430 and 460 nanometers) was nearly ten times more effective than red light (wavelengths between 630 and 680 nanometers) in producing a conductance of 15 centimoles per square meter per second. Stomata responded only slightly to green light. An action spectrum of stomatal responses to red light corresponded to that of CO(2) assimilation; the inhibitors of photosynthetic electron transport, cyanazine (2-chloro-4[1-cyano-1-methylethylamino]-6-ethylamino-s-triazine) and 3-(3,4-dichlorophenyl)-1,1-dimethylurea, eliminated the response to red light. This indicates that light absorption by chlorophyll is the cause of stomatal sensitivity to red light. Determination of flux response curves on leaves in the normal position (upper epidermis facing the light) or in the inverted position (lower epidermis facing the light) led to the conclusion that the photoreceptors for blue as well as for red light are located on or near the surfaces of the leaves; presumably they are in the guard cells themselves.

An ancestral stomatal patterning module revealed in the non-vascular land plant Physcomitrella patens

Science.gov (United States)

Chater, Caspar C.; Kamisugi, Yasuko

2016-01-01

The patterning of stomata plays a vital role in plant development and has emerged as a paradigm for the role of peptide signals in the spatial control of cellular differentiation. Research in Arabidopsis has identified a series of epidermal patterning factors (EPFs), which interact with an array of membrane-localised receptors and associated proteins (encoded by ERECTA and TMM genes) to control stomatal density and distribution. However, although it is well-established that stomata arose very early in the evolution of land plants, until now it has been unclear whether the established angiosperm stomatal patterning system represented by the EPF/TMM/ERECTA module reflects a conserved, universal mechanism in the plant kingdom. Here, we use molecular genetics to show that the moss Physcomitrella patens has conserved homologues of angiosperm EPF, TMM and at least one ERECTA gene that function together to permit the correct patterning of stomata and that, moreover, elements of the module retain function when transferred to Arabidopsis. Our data characterise the stomatal patterning system in an evolutionarily distinct branch of plants and support the hypothesis that the EPF/TMM/ERECTA module represents an ancient patterning system. PMID:27407102

Relationship between ammonia stomatal compensation point and nitrogen metabolism in arable crops: Current status of knowledge and potential modelling approaches

Energy Technology Data Exchange (ETDEWEB)

Massad, Raia Silvia [Institut National de la Recherche Agronomique (INRA), Environnement et Grandes Cultures, 78850 Thiverval-Grignon (France)], E-mail: massad@grignon.inra.fr; Loubet, Benjamin; Tuzet, Andree; Cellier, Pierre [Institut National de la Recherche Agronomique (INRA), Environnement et Grandes Cultures, 78850 Thiverval-Grignon (France)

2008-08-15

The ammonia stomatal compensation point of plants is determined by leaf temperature, ammonium concentration ([NH{sub 4}{sup +}]{sub apo}) and pH of the apoplastic solution. The later two depend on the adjacent cells metabolism and on leaf inputs and outputs through the xylem and phloem. Until now only empirical models have been designed to model the ammonia stomatal compensation point, except the model of Riedo et al. (2002. Coupling soil-plant-atmosphere exchange of ammonia with ecosystem functioning in grasslands. Ecological Modelling 158, 83-110), which represents the exchanges between the plant's nitrogen pools. The first step to model the ammonia stomatal compensation point is to adequately model [NH{sub 4}{sup +}]{sub apo}. This [NH{sub 4}{sup +}]{sub apo} has been studied experimentally, but there are currently no process-based quantitative models describing its relation to plant metabolism and environmental conditions. This study summarizes the processes involved in determining the ammonia stomatal compensation point at the leaf scale and qualitatively evaluates the ability of existing whole plant N and C models to include a model for [NH{sub 4}{sup +}]{sub apo}. - A model for ammonia stomatal compensation point at the leaf level scale was developed.

The biological effects of ozone on representative members of five groups of animal viruses

Energy Technology Data Exchange (ETDEWEB)

Bolton, D.C.; Zee, Y.C.; Osebold, J.W.

1982-04-01

In an effort to establish the biological relevance of the reactions of ozone with soluble proteins and lipid bilayer membrane systems, representative viruses from five major virus groups were exposed to moderate concentrations of ozone. The virus suspensions were exposed at 37/sup 0/C to 0.00, 0.16, and 0.64 ppm ozone in the gas phase. The ozone reacted with the virus suspensions as a thin film of fluid on the surface of a rotating culture bottle as the gas was drawn through the bottle at a flow rate of 2 liters/min. The three enveloped viruses tested exhibited different susceptibilities to ozone inactivation which correlated with their thermolability in the absence of ozone. The order of susceptibility to ozone inactivation of the enveloped viruses was vesicular stomatitis virus (VSV) (Rhabdoviridae) > influenza A virus (WSN strain) (Orthomyxoviridae) > infectious bovine rhinotracheitis virus (IBRV) (Herpesviridae). The inactivation reactions of the enveloped viruses with ozone showed pseudo-first-order kinetics. A simple reaction model was used to derive a reaction rate expression from which rate constrants and reaction stoichiometry were estimated. In contrast to the enveloped viruses, the two nonenveloped viruses examined were relatively resistant to ozone inactivation. Polio virus type I (Picornaviridae) was found to be completely resistant to ozone inactivation after 60 hr exposure to either ozone concentration, while infectious canine hepatitis virus (Adenoviridae) showed only slight inactivation after exposure to 0.64 ppm ozone for 66 hr. The significance of these results with regard to the reactions of ozone with cell membranes and other components is discussed.

Tree density and permafrost thaw depth influence water limitations on stomatal conductance in Siberian Arctic boreal forests

Science.gov (United States)

Kropp, H.; Loranty, M. M.; Natali, S.; Kholodov, A. L.; Alexander, H. D.; Zimov, N.

2017-12-01

Boreal forests may experience increased water stress under global climate change as rising air temperatures increase evaporative demand and decrease soil moisture. Increases in plant water stress can decrease stomatal conductance, and ultimately, decrease primary productivity. A large portion of boreal forests are located in Siberia, and are dominated by deciduous needleleaf trees, Larix spp. We investigated the variability and drivers of canopy stomatal conductance in upland Larix stands with different stand density that arose from differing fire severity. Our measurements focus on an open canopy stand with low tree density and deep permafrost thaw depth, and a closed canopy stand with high tree density and shallow permafrost thaw depth. We measured canopy stomatal conductance, soil moisture, and micrometeorological variables. Our results demonstrate that canopy stomatal conductance was significantly lower in the closed canopy stand with a significantly higher sensitivity to increases in atmospheric evaporative demand. Canopy stomatal conductance in both stands was tightly coupled to precipitation that occurred over the previous week; however, the closed canopy stand showed a significantly greater sensitivity to increases in precipitation compared to the open canopy stand. Differences in access to deep versus shallow soil moisture and the physical characteristics of the soil profile likely contribute to differences in sensitivity to precipitation between the two stands. Our results indicate that Larix primary productivity may be highly sensitive to changes in evaporative demand and soil moisture that can result of global climate change. However, the effect of increasing air temperatures and changes in precipitation will differ significantly depending on stand density, thaw depth, and the hydraulic characteristics of the soil profile.

[Comparison of protective properties of the smallpox DNA-vaccine based on the variola virus A30L gene and its variant with modified codon usage].

Science.gov (United States)

Maksiutov, R A; Shchelkunov, S N

2011-01-01

Efficacy of candidate DNA-vaccines based on the variola virus natural gene A30L and artificial gene A30Lopt with modified codon usage, optimized for expression in mammalian cells, was tested. The groups of mice were intracutaneously immunized three times with three-week intervals with candidate DNA-vaccines: pcDNA_A30L or pcDNA_A30Lopt, and in three weeks after the last immunization all mice in the groups were intraperitoneally infected by the ectromelia virus K1 strain in 10 LD50 dose for the estimation of protection. It was shown that the DNA-vaccines based on natural gene A30L and codon-optimized gene A30Lopt elicited virus, thereby neutralizing the antibody response and protected mice from lethal intraperitoneal challenge with the ectromelia virus with lack of statistically significant difference.

Natural variation in stomatal response to closing stimuli among Arabidopsis thaliana accessions after exposure to lowe VPD as a tool to recognize the mechanism of disturbed stomatal functioning

NARCIS (Netherlands)

Ali Niaei Fard, S.; Meeteren, van U.

2014-01-01

Stomatal responses to closing stimuli are disturbed after long-term exposure of plants to low vapour pressure deficit (VPD). The mechanism behind this disturbance is not fully understood. Genetic variation between naturally occurring ecotypes can be helpful to elucidate the mechanism controlling

An effective AIDS vaccine based on live attenuated vesicular stomatitis virus recombinants.

Science.gov (United States)

Rose, N F; Marx, P A; Luckay, A; Nixon, D F; Moretto, W J; Donahoe, S M; Montefiori, D; Roberts, A; Buonocore, L; Rose, J K

2001-09-07

We developed an AIDS vaccine based on attenuated VSV vectors expressing env and gag genes and tested it in rhesus monkeys. Boosting was accomplished using vectors with glycoproteins from different VSV serotypes. Animals were challenged with a pathogenic AIDS virus (SHIV89.6P). Control monkeys showed a severe loss of CD4+ T cells and high viral loads, and 7/8 progressed to AIDS with an average time of 148 days. All seven vaccinees were initially infected with SHIV89.6P but have remained healthy for up to 14 months after challenge with low or undetectable viral loads. Protection from AIDS was highly significant (p = 0.001). VSV vectors are promising candidates for human AIDS vaccine trials because they propagate to high titers and can be delivered without injection.

Computational fitness landscape for all gene-order permutations of an RNA virus.

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Kwang-il Lim

2009-02-01

Full Text Available How does the growth of a virus depend on the linear arrangement of genes in its genome? Answering this question may enhance our basic understanding of virus evolution and advance applications of viruses as live attenuated vaccines, gene-therapy vectors, or anti-tumor therapeutics. We used a mathematical model for vesicular stomatitis virus (VSV, a prototype RNA virus that encodes five genes (N-P-M-G-L, to simulate the intracellular growth of all 120 possible gene-order variants. Simulated yields of virus infection varied by 6,000-fold and were found to be most sensitive to gene-order permutations that increased levels of the L gene transcript or reduced levels of the N gene transcript, the lowest and highest expressed genes of the wild-type virus, respectively. Effects of gene order on virus growth also depended upon the host-cell environment, reflecting different resources for protein synthesis and different cell susceptibilities to infection. Moreover, by computationally deleting intergenic attenuations, which define a key mechanism of transcriptional regulation in VSV, the variation in growth associated with the 120 gene-order variants was drastically narrowed from 6,000- to 20-fold, and many variants produced higher progeny yields than wild-type. These results suggest that regulation by intergenic attenuation preceded or co-evolved with the fixation of the wild type gene order in the evolution of VSV. In summary, our models have begun to reveal how gene functions, gene regulation, and genomic organization of viruses interact with their host environments to define processes of viral growth and evolution.

Impaired Malate and Fumarate Accumulation Due to the Mutation of the Tonoplast Dicarboxylate Transporter Has Little Effects on Stomatal Behavior.

Science.gov (United States)

Medeiros, David B; Barros, Kallyne A; Barros, Jessica Aline S; Omena-Garcia, Rebeca P; Arrivault, Stéphanie; Sanglard, Lílian M V P; Detmann, Kelly C; Silva, Willian Batista; Daloso, Danilo M; DaMatta, Fábio M; Nunes-Nesi, Adriano; Fernie, Alisdair R; Araújo, Wagner L

2017-11-01

Malate is a central metabolite involved in a multiplicity of plant metabolic pathways, being associated with mitochondrial metabolism and playing significant roles in stomatal movements. Vacuolar malate transport has been characterized at the molecular level and is performed by at least one carrier protein and two channels in Arabidopsis ( Arabidopsis thaliana ) vacuoles. The absence of the Arabidopsis tonoplast Dicarboxylate Transporter (tDT) in the tdt knockout mutant was associated previously with an impaired accumulation of malate and fumarate in leaves. Here, we investigated the consequences of this lower accumulation on stomatal behavior and photosynthetic capacity as well as its putative metabolic impacts. Neither the stomatal conductance nor the kinetic responses to dark, light, or high CO 2 were highly affected in tdt plants. In addition, we did not observe any impact on stomatal aperture following incubation with abscisic acid, malate, or citrate. Furthermore, an effect on photosynthetic capacity was not observed in the mutant lines. However, leaf mitochondrial metabolism was affected in the tdt plants. Levels of the intermediates of the tricarboxylic acid cycle were altered, and increases in both light and dark respiration were observed. We conclude that manipulation of the tonoplastic organic acid transporter impacted mitochondrial metabolism, while the overall stomatal and photosynthetic capacity were unaffected. © 2017 American Society of Plant Biologists. All Rights Reserved.

Viral fusion efficacy of specific H3N2 influenza virus reassortant combinations at single-particle level

Science.gov (United States)

Hsu, Hung-Lun; Millet, Jean K.; Costello, Deirdre A.; Whittaker, Gary R.; Daniel, Susan

2016-01-01

Virus pseudotyping is a useful and safe technique for studying entry of emerging strains of influenza virus. However, few studies have compared different reassortant combinations in pseudoparticle systems, or compared entry kinetics of native viruses and their pseudotyped analogs. Here, vesicular stomatitis virus (VSV)-based pseudovirions displaying distinct influenza virus envelope proteins were tested for fusion activity. We produced VSV pseudotypes containing the prototypical X-31 (H3) HA, either alone or with strain-matched or mismatched N2 NAs. We performed single-particle fusion assays using total internal reflection fluorescence microscopy to compare hemifusion kinetics among these pairings. Results illustrate that matching pseudoparticles behaved very similarly to native virus. Pseudoparticles harboring mismatched HA-NA pairings fuse at significantly slower rates than native virus, and NA-lacking pseudoparticles exhibiting the slowest fusion rates. Relative viral membrane HA density of matching pseudoparticles was higher than in mismatching or NA-lacking pseudoparticles. An equivalent trend of HA expression level on cell membranes of HA/NA co-transfected cells was observed and intracellular trafficking of HA was affected by NA co-expression. Overall, we show that specific influenza HA-NA combinations can profoundly affect the critical role played by HA during entry, which may factor into viral fitness and the emergence of new pandemic influenza viruses. PMID:27752100

Influence of stomatic aperture on photosynthetic activity of bean-seedlings leaves

International Nuclear Information System (INIS)

Suarez Moya, J.; Fernandez Gonzalez, J.

1984-01-01

The present paper contains the data of photosynthetic activity and stomatic aperture of bean-seedlings Ieaves, and the relations obtained with both results. It has been observed that the product of photosynthetic activity by the resistance; to transpiration measured by a promoter ia a constant, between some limits. (Author) 45 refs

Drought limitations to leaf-level gas exchange: results from a model linking stomatal optimization and cohesion-tension theory

Science.gov (United States)

Kimberly A. Novick; Chelcy F. Miniat; James M. Vose

2016-01-01

We merge concepts from stomatal optimization theory and cohesion–tension theory to examine the dynamics of three mechanisms that are potentially limiting to leaf-level gas exchange in trees during drought: (1) a ‘demand limitation’ driven by an assumption of optimal stomatal functioning; (2) ‘hydraulic limitation’ of water movement from the roots to the leaves...

Oncolytic Viruses-Interaction of Virus and Tumor Cells in the Battle to Eliminate Cancer.

Science.gov (United States)

Howells, Anwen; Marelli, Giulia; Lemoine, Nicholas R; Wang, Yaohe

2017-01-01

Oncolytic viruses (OVs) are an emerging treatment option for many cancer types and have recently been the focus of extensive research aiming to develop their therapeutic potential. The ultimate aim is to design a virus which can effectively replicate within the host, specifically target and lyse tumor cells and induce robust, long lasting tumor-specific immunity. There are a number of viruses which are either naturally tumor-selective or can be modified to specifically target and eliminate tumor cells. This means they are able to infect only tumor cells and healthy tissue remains unharmed. This specificity is imperative in order to reduce the side effects of oncolytic virotherapy. These viruses can also be modified by various methods including insertion and deletion of specific genes with the aim of improving their efficacy and safety profiles. In this review, we have provided an overview of the various virus species currently being investigated for their oncolytic potential and the positive and negative effects of a multitude of modifications used to increase their infectivity, anti-tumor immunity, and treatment safety, in particular focusing on the interaction of tumor cells and OVs.

Differential Effects of Phosphatidylinositol 4-Kinase (PI4K and 3-Kinase (PI3K Inhibitors on Stomatal Responses to Environmental Signals

Directory of Open Access Journals (Sweden)

Koh Iba

2017-05-01

Full Text Available Specific cellular components including products of phosphatidylinositol (PI metabolism play an important role as signaling molecules in stomatal responses to environmental signals. In this study, pharmacological inhibitors of a set of cellular components, including PI4-kinase (PI4K and PI3K, were used to investigate stomatal closure in response to CO2, darkness, and abscisic acid (ABA. Treatment with PAO, a specific inhibitor of PI4K, specifically inhibited the stomatal response to CO2 compared with that to darkness and ABA. In contrast, treatment with LY294002, a PI3K-specific inhibitor, specifically inhibited the stomatal response to darkness compared with that to CO2 and ABA. The specific inhibitory effects of PAO and LY294002 were also observed as changes in the spatial density of dot-like structures labeled by green fluorescent protein-tagged PATROL1, a protein that controls stomatal aperture possibly via regulation of H+-ATPase amount in guard cell plasma membranes. Our results suggest an important role for PI4K and PI3K in the CO2 and darkness signal transduction pathways, respectively, that mediate PATROL1 dynamics.

Antiviral activity of gliotoxin, gentian violet and brilliant green against Nipah and Hendra virus in vitro

Directory of Open Access Journals (Sweden)

Meyer Adam G

2009-11-01

Full Text Available Abstract Background Using a recently described monolayer assay amenable to high throughput screening format for the identification of potential Nipah virus and Hendra virus antivirals, we have partially screened a low molecular weight compound library (>8,000 compounds directly against live virus infection and identified twenty eight promising lead molecules. Initial single blind screens were conducted with 10 μM compound in triplicate with a minimum efficacy of 90% required for lead selection. Lead compounds were then further characterised to determine the median efficacy (IC50, cytotoxicity (CC50 and the in vitro therapeutic index in live virus and pseudotype assay formats. Results While a number of leads were identified, the current work describes three commercially available compounds: brilliant green, gentian violet and gliotoxin, identified as having potent antiviral activity against Nipah and Hendra virus. Similar efficacy was observed against pseudotyped Nipah and Hendra virus, vesicular stomatitis virus and human parainfluenza virus type 3 while only gliotoxin inhibited an influenza A virus suggesting a non-specific, broad spectrum activity for this compound. Conclusion All three of these compounds have been used previously for various aspects of anti-bacterial and anti-fungal therapy and the current results suggest that while unsuitable for internal administration, they may be amenable to topical antiviral applications, or as disinfectants and provide excellent positive controls for future studies.

Effect of Light Quality on Stomatal Opening in Leaves of Xanthium strumarium L. 1

Science.gov (United States)

Sharkey, Thomas D.; Raschke, Klaus

1981-01-01

Flux response curves were determined at 16 wavelengths of light for the conductance for water vapor of the lower epidermis of detached leaves of Xanthium strumarium L. An action spectrum of stomatal opening resulted in which blue light (wavelengths between 430 and 460 nanometers) was nearly ten times more effective than red light (wavelengths between 630 and 680 nanometers) in producing a conductance of 15 centimoles per square meter per second. Stomata responded only slightly to green light. An action spectrum of stomatal responses to red light corresponded to that of CO2 assimilation; the inhibitors of photosynthetic electron transport, cyanazine (2-chloro-4[1-cyano-1-methylethylamino]-6-ethylamino-s-triazine) and 3-(3,4-dichlorophenyl)-1,1-dimethylurea, eliminated the response to red light. This indicates that light absorption by chlorophyll is the cause of stomatal sensitivity to red light. Determination of flux response curves on leaves in the normal position (upper epidermis facing the light) or in the inverted position (lower epidermis facing the light) led to the conclusion that the photoreceptors for blue as well as for red light are located on or near the surfaces of the leaves; presumably they are in the guard cells themselves. PMID:16662069

Identifying the tundra-forest border in the stomate record: an analysis of lake surface samples from the Yellowknife area, Northwest Territories, Canada

Energy Technology Data Exchange (ETDEWEB)

Hansen, B.C.S. [Minnesota Univ., Minneapolis, MN (United States). Limnological Research Center; MacDonald, G.M. [California Univ., Los Angeles, CA (United States). Dept. of Botanical Sciences; Moser, K.A. [McMaster Univ., Hamilton, ON (Canada)

1996-05-01

The relationship between conifer stomata and existing vegetation across tundra, forest-tundra, and closed zones in the Yellowknife area of the Northwest Territories was studied. Conifer stomata were identified in surface samples from lakes in the treeline zone, but were absent in samples from tundra lakes. Stomate analysis was recorded and the results were presented in a concentration diagram plotting stomate concentrations according to vegetation zone. Conifer stomate analysis was not able to resolve differences between forest-tundra and closed forest. Nevertheless, it was suggested that stomate analysis will become an important technique supplementing pollen analysis for reconstructing past tree-line changes since the presence of stomata in lakes make it possible to separate the tundra from forest-tundra and closed forest. The limited dispersal of conifer stomata permitted a better resolution of tree-line boundaries than did pollen. 13 refs., 3 figs.

Investigation of the effect of the rebamipide mouthwash on the crisis of the stomatitis induced by the cancer chemotherapy and/or radiotherapy

International Nuclear Information System (INIS)

Kawata, Keishi; Hanawa, Takehisa; Hanawa, Kazumi

2001-01-01

Stomatitis is well-known as one of the undesirable side effects induced by high and/or multiple dosing of cytotoxic drugs such as 5-fluorouracil (5-FU). Stomatitis causes pain in the oral cavity, impaired swallowing or loss of appetite, and finally, lowering of the quality of life (QOL) of patients. In this study, we attempted to apply a new mouthwash containing rebamipide (REB) which is known as the anti-activated oxygen agent. Rebamipide mouthwash (REB-M) showed the effectiveness to the crisis of the stomatitis during the cancer chemotherapy and/or radiotherapy. (author)

Nitric Oxide (NO) Measurements in Stomatal Guard Cells.

Science.gov (United States)

Agurla, Srinivas; Gayatri, Gunja; Raghavendra, Agepati S

2016-01-01

The quantitative measurement of nitric oxide (NO) in plant cells acquired great importance, in view of the multifaceted function and involvement of NO as a signal in various plant processes. Monitoring of NO in guard cells is quite simple because of the large size of guard cells and ease of observing the detached epidermis under microscope. Stomatal guard cells therefore provide an excellent model system to study the components of signal transduction. The levels and functions of NO in relation to stomatal closure can be monitored, with the help of an inverted fluorescence or confocal microscope. We can measure the NO in guard cells by using flouroprobes like 4,5-diamino fluorescein diacetate (DAF-2DA). This fluorescent dye, DAF-2DA, is cell permeable and after entry into the cell, the diacetate group is removed by the cellular esterases. The resulting DAF-2 form is membrane impermeable and reacts with NO to generate the highly fluorescent triazole (DAF-2T), with excitation and emission wavelengths of 488 and 530 nm, respectively. If time-course measurements are needed, the epidermis can be adhered to a cover-glass or glass slide and left in a small petri dishes. Fluorescence can then be monitored at required time intervals; with a precaution that excitation is done minimally, only when a fluorescent image is acquired. The present method description is for the epidermis of Arabidopsis thaliana and Pisum sativum and should work with most of the other dicotyledonous plants.

Felix Hoppe-Seyler Lecture 1997. Protective antibody responses against viruses.

Science.gov (United States)

Zinkernagel, R M

1997-08-01

Neutralizing antibody responses against the acute cytopathic vesicular stomatitis virus (VSV) have been studied in mice to evaluate their general characteristics including specificity, self-/non-self discrimination and memory. IgM responses are generated very early, by day 3 to 4, in a T helper cell-independent fashion and without VSV having polyclonal activating capacities. The order of the glycoprotein tips on the virus envelope (multiple, 8-10 nm distance, paracrystalline) exhibiting the neutralizing determinants are key to this prompt response. These paracrystalline identical multimeric antigens are characteristic of infectious agents and are always reacted against by B cells. Self-antigens that are accessible to B cells in the intact host are either monomeric in serum or mobile multimers on cell surfaces; these configurations need contact dependent or contact independent T help, respectively. Because T help is tolerant against self-antigens, no anti-self B cell responses are usually induced against monomeric self-antigens. If collagen or DNA (rigid multimeric self-antigens) become accessible, however, they may become targets of auto-antibody responses. The antibody repertoire against VSV is partially contained in the germline and partially is generated by somatic mutation; they seem not to undergo affinity-maturation. In any case protection against lethal infection is dependent upon strictly T helper cell dependent IgG generated by day 6 to 7 and reaches a protective level of about 1-10 micrograms/ml. Interesting affinity/avidity and onrate above a minimal threshold are of no apparent advantage for protection in vivo. Maintenance of these antibody levels by antigen depots, and not the presence of memory B cells alone, is key to providing protective immunological memory. Collectively these data suggest that studying biologically important protective antibody responses may modify some of the parameters that have been defined by studying hapten specific antibody

Stomatal and pavement cell density linked to leaf internal CO2 concentration

Czech Academy of Sciences Publication Activity Database

Šantrůček, Jiří; Vráblová, M.; Šimková, Marie; Hronková, Marie; Drtinová, M.; Květoň, J.; Vrábl, D.; Kubásek, J.; Macková, J.; Wiesnerová, Dana; Neuwithová, J.; Schreiber, L.

2014-01-01

Roč. 114, č. 2 (2014), s. 191-202 ISSN 0305-7364 R&D Projects: GA ČR(CZ) GAP501/12/1261 Institutional support: RVO:60077344 Keywords : Stomatal density * Stomata development * Pavement cells Subject RIV: CE - Biochemistry Impact factor: 3.654, year: 2014

Antiviral activity of Petiveria alliacea against the bovine viral diarrhea virus.

Science.gov (United States)

Ruffa, M J; Perusina, M; Alfonso, V; Wagner, M L; Suriano, M; Vicente, C; Campos, R; Cavallaro, L

2002-07-01

Natural products are a relevant source of antiviral drugs. Five medicinal plants used in Argentina have been assayed to detect inhibition of viral growth. Antiviral activity of the infusions and methanolic extracts of Aristolochia macroura, Celtis spinosa, Plantago major, Schinus areira, Petiveria alliacea and four extracts obtained from the leaves and stems of the last plant were evaluated by the plaque assay. P. alliacea, unlike A. macroura, C. spinosa, P. major and S. areira, inhibited bovine viral diarrhea virus (BVDV) replication. Neither P. alliacea nor the assays of the other plants were active against herpes simplex virus type 1, poliovirus type 1, adenovirus serotype 7 and vesicular stomatitis virus type 1. Four extracts of P. alliacea were assayed to detect anti-BVDV activity. Ethyl acetate (EC(50) of 25 microg/ml) and dichloromethane (EC(50) of 43 microg/ml) extracts were active; moreover, promising SI (IC(50)/EC(50)) values were obtained. BVDV is highly prevalent in the cattle population, there are no antiviral compounds available; additionally, it is a viral model of the hepatitis C virus. For these reasons and in view of the results obtained, the isolation and characterization of the antiviral components present in the P. alliacea extracts is worth carrying out in the future. Copyright 2002 S. Karger AG, Basel

Bat airway epithelial cells: a novel tool for the study of zoonotic viruses.

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Isabella Eckerle

Full Text Available Bats have been increasingly recognized as reservoir of important zoonotic viruses. However, until now many attempts to isolate bat-borne viruses in cell culture have been unsuccessful. Further, experimental studies on reservoir host species have been limited by the difficulty of rearing these species. The epithelium of the respiratory tract plays a central role during airborne transmission, as it is the first tissue encountered by viral particles. Although several cell lines from bats were established recently, no well-characterized, selectively cultured airway epithelial cells were available so far. Here, primary cells and immortalized cell lines from bats of the two important suborders Yangochiroptera and Yinpterochiroptera, Carollia perspicillata (Seba's short-tailed bat and Eidolon helvum (Straw-colored fruit bat, were successfully cultured under standardized conditions from both fresh and frozen organ specimens by cell outgrowth of organ explants and by the use of serum-free primary cell culture medium. Cells were immortalized to generate permanent cell lines. Cells were characterized for their epithelial properties such as expression of cytokeratin and tight junctions proteins and permissiveness for viral infection with Rift-Valley fever virus and vesicular stomatitis virus Indiana. These cells can serve as suitable models for the study of bat-borne viruses and complement cell culture models for virus infection in human airway epithelial cells.

Suramin is a potent inhibitor of Chikungunya and Ebola virus cell entry.

Science.gov (United States)

Henß, Lisa; Beck, Simon; Weidner, Tatjana; Biedenkopf, Nadine; Sliva, Katja; Weber, Christopher; Becker, Stephan; Schnierle, Barbara S

2016-08-31

Chikungunya virus (CHIKV) is a mosquito-transmitted alphavirus that causes high fever, rash, and recurrent arthritis in humans. It has efficiently adapted to Aedes albopictus, which also inhabits temperate regions and currently causes large outbreaks in the Caribbean and Latin America. Ebola virus (EBOV) is a member of the filovirus family. It causes the Ebola virus disease (EDV), formerly known as Ebola hemorrhagic fever in humans and has a mortality rate of up to 70 %. The last outbreak in Western Africa was the largest in history and has caused approximately 25,000 cases and 10,000 deaths. For both viral infections no specific treatment or licensed vaccine is currently available. The bis-hexasulfonated naphthylurea, suramin, is used as a treatment for trypanosome-caused African river blindness. As a competitive inhibitor of heparin, suramin has been described to have anti-viral activity. We tested the activity of suramin during CHIKV or Ebola virus infection, using CHIKV and Ebola envelope glycoprotein pseudotyped lentiviral vectors and wild-type CHIKV and Ebola virus. Suramin efficiently inhibited CHIKV and Ebola envelope-mediated gene transfer while vesicular stomatitis virus G protein pseudotyped vectors were only marginally affected. In addition, suramin was able to inhibit wild-type CHIKV and Ebola virus replication in vitro. Inhibition occurred at early time points during CHIKV infection. Suramin, also known as Germanin or Bayer-205, is a market-authorized drug, however shows significant side effects, which probably prevents its use as a CHIKV drug, but due to the high lethality of Ebola virus infections, suramin might be valuable against Ebola infections.

Estimating Latest Cretaceous and Tertiary Atmospheric PCO2 from Stomatal Indices

Science.gov (United States)

Royer, D. L.; Wing, S. L.; Beerling, D. J.

2001-05-01

Most modern C3 seed plants show an inverse relationship between PCO2 and stomatal index (SI), where SI is the proportion of epidermal cells that are stomatal packages. This plant-atmosphere response therefore provides a reliable approach for estimating paleo-CO2 levels. Since stomatal responses to CO2 are generally species-specific, one is limited in paleo-reconstructions to species that exist both in the fossil record and living today. Fossils morphologically similar to living Ginkgo biloba and Metasequoia glyptostroboides extend back to the early and late Cretaceous, respectively, indicating that the fossil and living forms are very closely related. Measurements of SI made on fossil Ginkgo and Metasequoia were calibrated with historical collections of G. biloba and M. glyptostroboides leaves from sites that developed during the anthropogenically-driven CO2 increases of the past 145 years (288-369 ppmv) and with saplings of G. biloba and M. glyptostroboides grown in CO2 controlled growth chambers (350-800 ppmv). Both nonlinear regressions are highly significant (Ginkgo: n = 40, r2 = 0.91; Metasequoia: n = 18; r2 = 0.85). Results from a sequence of 23 latest Cretaceous to early Eocene-aged Ginkgo-bearing sites indicate that CO2 remained between 300 and 450 ppmv with the exception of one high estimate ( ~800 ppmv) near the Paleocene/Eocene boundary, and results from 4 middle Miocene-aged Ginkgo- and Metasequoia-bearing sites indicate that CO2 was between 320 and 400 ppmv. If correct, the CO2 values estimated here are too low to explain via the CO2 greenhouse effect alone the higher global mean temperatures (e.g., 3-4 ° C for the early Eocene) inferred from models and geological data for these two intervals.

Selective elimination of HIV-1-infected cells by Env-directed, HIV-1-based virus-like particles

International Nuclear Information System (INIS)

Peretti, Silvia; Schiavoni, Ilaria; Pugliese, Katherina; Federico, Maurizio

2006-01-01

We recently showed that both replicating and resting cells cultivated with ganciclovir (GCV) were killed when challenged with vesicular stomatitis virus G glycoprotein pseudotyped HIV-1-based virus-like particles (VLPs) carrying the Nef7 (i.e., an HIV-1 Nef mutant incorporating in virions at high levels)/herpes simplex virus-1 thymidine kinase (HSV-TK) fusion product. On this basis, a novel anti-HIV therapeutic approach based on Nef7/TK VLPs expressing X4 or R5 HIV cell receptor complexes has been attempted. We here report that (CD4-CXCR4) and (CD4-CCR5) Nef7-based VLPs efficiently enter cells infected by X4- or R5-tropic HIV-1 strains, respectively. Importantly, the delivery of the VLP-associated Nef7/TK led to cell death upon GCV treatment. Of interest, VLPs were effective also against non-replicating, HIV-1-infected primary human monocyte-derived macrophages. HIV-targeted VLPs represent a promising candidate for the treatment of persistently HIV-1-infected cells that are part of virus reservoirs resistant to HAART therapies

Linking stomatal traits and expression of slow anion channel genes HvSLAH1 2 HvSLAC1 with grain yield for increasing salinity tolerance in barley

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Xiaohui eLiu

2014-11-01

Full Text Available Soil salinity is an environmental and agricultural problem in many parts of the world. One of the keys to breeding barley for adaptation to salinity lies in a better understanding of the genetic control of stomatal regulation. We have employed a range of physiological and molecular techniques (stomata assay, gas exchange, phylogenetic analysis, QTL analysis, and gene expression to investigate stomatal behaviour and genotypic variation in barley cultivars and a genetic population in four experimental trials. A set of relatively efficient and reliable methods were developed for the characterisation of stomatal behaviour of large numbers of varieties and genetic lines. Furthermore, we have found a large genetic variation of gas exchange and stomatal traits in barley in response to salinity stress. Salt-tolerant CM72 showed significantly larger stomatal aperture in 200 mM NaCl treatment than that of salt-sensitive Gairdner. Stomatal traits such as aperture width/length were found to significantly correlate with grain yield in salt treatment. Phenotypic characterisation and QTL analysis of a segregating double haploid population of the CM72/Gairdner resulted in the identification of significant stomatal traits-related QTLs for salt tolerance. Moreover, expression analysis of the slow anion channel genes HvSLAH1 and HvSLAC1 demonstrated that their up-regulation is linked to high barley grain yield in the field.

Recalibrating the Ginkgo Stomatal Index Proxy for Past CO2 with Herbarium Specimens

Science.gov (United States)

Conde, G. D.; Retallack, G.

2015-12-01

The stomatal index of plant cuticles is inversely related to atmospheric CO2 concentrations, as calibrated from greenhouse experiments and herbarium specimens. Such calibration data for Ginkgo biloba are available for the ongoing rise in atmospheric CO2 and for high levels of CO2 anticipated in the future, but lacking for low CO2 levels of preindustrial and glacial ages. The oldest modern specimen that we have been able to obtain consists of leaf fragments collected in 1829 and provided by Arne Anderberg from the Swedish Natural History Museum. The specimen was labeled "Argentina", but also "Hortus Botanicus Augustinus", a garden founded in 1638 in Amsterdam, Netherlands. Ginkgo has a very thin cuticle that is difficult to prepare, but images very similar to cuticular preparation can be obtained by backscatter SEM imagery. We also obtained secondary SEM images of the same areas and counted 13 images with 6,184 cells from five leaf fragments. Our analyses yield a stomatal index of 10.9 ± 0.9 % for an atmospheric CO2 of 286 ppm, as determined by ice core data from Ciais and Sabine for IPCC-2013. This value is lower than from previous calibration curves for this level of CO2 and changes their curvature. With additional late nineteenth, twentieth and twenty-first century leaves, we can improve both the precision and lower limits of the transfer function for atmospheric CO2 from Ginkgo stomatal index last revised in 2009.

[Validation of the modified algorithm for predicting host susceptibility to viruses taking into account susceptibility parameters of primary target cell cultures and natural immunity factors].

Science.gov (United States)

Zhukov, V A; Shishkina, L N; Safatov, A S; Sergeev, A A; P'iankov, O V; Petrishchenko, V A; Zaĭtsev, B N; Toporkov, V S; Sergeev, A N; Nesvizhskiĭ, Iu V; Vorob'ev, A A

2010-01-01

The paper presents results of testing a modified algorithm for predicting virus ID50 values in a host of interest by extrapolation from a model host taking into account immune neutralizing factors and thermal inactivation of the virus. The method was tested for A/Aichi/2/68 influenza virus in SPF Wistar rats, SPF CD-1 mice and conventional ICR mice. Each species was used as a host of interest while the other two served as model hosts. Primary lung and trachea cells and secretory factors of the rats' airway epithelium were used to measure parameters needed for the purpose of prediction. Predicted ID50 values were not significantly different (p = 0.05) from those experimentally measured in vivo. The study was supported by ISTC/DARPA Agreement 450p.

Production of Antigens and Antibodies for Diagnosis of Arbovirus Diseases.

Science.gov (United States)

1994-05-20

for Germiston, Qalyub, Sicilian, vesicular stomatitis Indiana, and Ganjam viruses. The antigens were inactivated with beta-propiolactone. Rabbits were...vesicular stomatitis Indiana, and Ganjam viruses. The antigens were inactivated with beta-propiolactone. Rabbits were immunized successfully intravenously...370 sm4 6 229 Sicilian Sabin sm37,Vero2 1 23 VS-Indiana Ind. Lab sm7 1 45 Ganjam IG 619 sm5 1 67 Additionally, 22 viruses were passaged in baby mice

The Arabidopsis homolog of human G3BP1 is a key regulator of stomatal and apoplastic immunity

KAUST Repository

Abulfaraj, Aala A.; Mariappan, Kiruthiga; Bigeard, Jean; Manickam, Prabhu; Blilou, Ikram; Guo, Xiujie; Al-Babili, Salim; Pflieger, Delphine; Hirt, Heribert; Rayapuram, Naganand

2018-01-01

Mammalian Ras-GTPase–activating protein SH3-domain–binding proteins (G3BPs) are a highly conserved family of RNA-binding proteins that link kinase receptor-mediated signaling to RNA metabolism. Mammalian G3BP1 is a multifunctional protein that functions in viral immunity. Here, we show that the Arabidopsis thaliana homolog of human G3BP1 negatively regulates plant immunity. Arabidopsis g3bp1 mutants showed enhanced resistance to the virulent bacterial pathogen Pseudomonas syringae pv. tomato. Pathogen resistance was mediated in Atg3bp1 mutants by altered stomatal and apoplastic immunity. Atg3bp1 mutants restricted pathogen entry into stomates showing insensitivity to bacterial coronatine–mediated stomatal reopening. AtG3BP1 was identified as a negative regulator of defense responses, which correlated with moderate up-regulation of salicylic acid biosynthesis and signaling without growth penalty.

The Arabidopsis homolog of human G3BP1 is a key regulator of stomatal and apoplastic immunity

KAUST Repository

Abulfaraj, Aala Abdulaziz Hussien

2018-05-31

Mammalian Ras-GTPase–activating protein SH3-domain–binding proteins (G3BPs) are a highly conserved family of RNA-binding proteins that link kinase receptor-mediated signaling to RNA metabolism. Mammalian G3BP1 is a multifunctional protein that functions in viral immunity. Here, we show that the Arabidopsis thaliana homolog of human G3BP1 negatively regulates plant immunity. Arabidopsis g3bp1 mutants showed enhanced resistance to the virulent bacterial pathogen Pseudomonas syringae pv. tomato. Pathogen resistance was mediated in Atg3bp1 mutants by altered stomatal and apoplastic immunity. Atg3bp1 mutants restricted pathogen entry into stomates showing insensitivity to bacterial coronatine–mediated stomatal reopening. AtG3BP1 was identified as a negative regulator of defense responses, which correlated with moderate up-regulation of salicylic acid biosynthesis and signaling without growth penalty.

RNAi-directed downregulation of vacuolar H(+ -ATPase subunit a results in enhanced stomatal aperture and density in rice.

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Huiying Zhang

Full Text Available Stomatal movement plays a key role in plant development and response to drought and salt stress by regulating gas exchange and water loss. A number of genes have been demonstrated to be involved in the regulation of this process. Using inverse genetics approach, we characterized the function of a rice (Oryza sativa L. vacuolar H(+-ATPase subunit A (OsVHA-A gene in stomatal conductance regulation and physiological response to salt and osmotic stress. OsVHA-A was constitutively expressed in different rice tissues, and the fusion protein of GFP-OsVHA-A was exclusively targeted to tonoplast when transiently expressed in the onion epidermal cells. Heterologous expression of OsVHA-A was able to rescue the yeast mutant vma1Δ (lacking subunit A activity phenotype, suggesting that it partially restores the activity of V-ATPase. Meanwhile, RNAi-directed knockdown of OsVHA-A led to a reduction of vacuolar H(+-ATPase activity and an enhancement of plasma membrane H(+-ATPase activity, thereby increasing the concentrations of extracellular H(+ and intracellular K(+ and Na(+ under stress conditions. Knockdown of OsVHA-A also resulted in the upregulation of PAM3 (plasma membrane H(+-ATPase 3 and downregulation of CAM1 (calmodulin 1, CAM3 (calmodulin 3 and YDA1 (YODA, a MAPKK gene. Altered level of the ion concentration and the gene expression by knockdown of OsVHA-A probably resulted in expanded aperture of stomatal pores and increased stomatal density. In addition, OsVHA-A RNAi plants displayed significant growth inhibition under salt and osmotic stress conditions. Taken together, our results suggest that OsVHA-A takes part in regulating stomatal density and opening via interfering with pH value and ionic equilibrium in guard cells and thereby affects the growth of rice plants.

Internal coordination between hydraulics and stomatal control in leaves.

Science.gov (United States)

Brodribb, Tim J; Jordan, Gregory J

2008-11-01

The stomatal response to changing leaf-atmospheric vapour pressure gradient (D(l)) is a crucial yet enigmatic process that defines the daily course of leaf gas exchange. Changes in the hydration of epidermal cells are thought to drive this response, mediated by the transpiration rate and hydraulic conductance of the leaf. Here, we examine whether species-specific variation in the sensitivity of leaves to perturbation of D(l) is related to the efficiency of water transport in the leaf (leaf hydraulic conductivity, K(leaf)). We found good correlation between maximum liquid (K(leaf)) and gas phase conductances (g(max)) in leaves, but there was no direct correlation between normalized D(l) sensitivity and K(leaf). The impact of K(leaf) on D(l) sensitivity in our diverse sample of eight species was important only after accounting for the strong relationship between K(leaf) and g(max). Thus, the ratio of g(max)/K(leaf) was strongly correlated with stomatal sensitivity to D(l). This ratio is an index of the degree of hydraulic buffering of the stomata against changes in D(l), and species with high g(max) relative to K(leaf) were the most sensitive to D(l) perturbation. Despite the potentially high adaptive significance of this phenomenon, we found no significant phylogenetic or ecological trend in our species.

Quantitative Evaluation of Stomatal Cytoskeletal Patterns during the Activation of Immune Signaling in Arabidopsis thaliana.

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Masaki Shimono

Full Text Available Historically viewed as primarily functioning in the regulation of gas and water vapor exchange, it is now evident that stomata serve an important role in plant immunity. Indeed, in addition to classically defined functions related to cell architecture and movement, the actin cytoskeleton has emerged as a central component of the plant immune system, underpinning not only processes related to cell shape and movement, but also receptor activation and signaling. Using high resolution quantitative imaging techniques, the temporal and spatial changes in the actin microfilament array during diurnal cycling of stomatal guard cells has revealed a highly orchestrated transition from random arrays to ordered bundled filaments. While recent studies have demonstrated that plant stomata close in response to pathogen infection, an evaluation of stimulus-induced changes in actin cytoskeletal dynamics during immune activation in the guard cell, as well as the relationship of these changes to the function of the actin cytoskeleton and stomatal aperture, remains undefined. In the current study, we employed quantitative cell imaging and hierarchical clustering analyses to define the response of the guard cell actin cytoskeleton to pathogen infection and the elicitation of immune signaling. Using this approach, we demonstrate that stomatal-localized actin filaments respond rapidly, and specifically, to both bacterial phytopathogens and purified pathogen elicitors. Notably, we demonstrate that higher order temporal and spatial changes in the filament array show distinct patterns of organization during immune activation, and that changes in the naïve diurnal oscillations of guard cell actin filaments are perturbed by pathogens, and that these changes parallel pathogen-induced stomatal gating. The data presented herein demonstrate the application of a highly tractable and quantifiable method to assign transitions in actin filament organization to the activation of

Modern pollen and stomate deposition in lake surface sediments from across the treeline on the Kola Peninsula, Russia.

Science.gov (United States)

Gervais, B R.; MacDonald, G M.

2001-04-01

We sampled and analyzed surface sediments from 31 lakes along a latitudinal transect crossing the coniferous treeline on the Kola Peninsula, Russia. The major vegetation zones along the transect were tundra, birch-forest tundra, pine-forest tundra, and forest. The results indicate that the major vegetation types in our study area have distinct pollen spectra. Sum-of-squares cluster analysis and principal components analysis (PCA) groupings of pollen sites correspond to the major vegetation zones. PCA ordination of taxa indicates that the first axis separates taxa typical of the forest zone (Pinus, Picea) from taxa typical of tundra and forest-tundra zones (Polypodiaceae, Ericaceae, and Betula). The current position of the coniferous treeline, defined in our region by Pinus sylvestris, occurs roughly where Pinus pollen values reach 35% or greater. Arboreal pollen (AP)/non-arboreal pollen (NAP) ratios were calculated for each site and plotted against geographic distance along the transect. AP/NAP ratios of 7 or greater are found within pine-forest tundra and forest vegetation zones. Pinus stomates (dispersed stomatal guard cells) are absent from sites north of the coniferous treeline and all but two samples from the forested sites contain stomates. Stomate concentrations among the samples are highly variable and range from 10 to 458 per ml and positively correlate with the changing Pinus pollen values.

Acquired changes in stomatal characteristics in response to ozone during plant growth and leaf development of bush beans (Phaseolus vulgaris L.) indicate phenotypic plasticity

International Nuclear Information System (INIS)

Elagoez, Vahram; Han, Susan S.; Manning, William J.

2006-01-01

Bush bean (Phaseolus vulgaris L.) lines 'S156' (O 3 -sensitive)/'R123' (O 3 -tolerant) and cultivars 'BBL 290' (O 3 -sensitive)/'BBL 274' (O 3 -tolerant) were used to study the effects of O 3 on stomatal conductance (g s ), density, and aperture size on leaf and pod surfaces with the objective of establishing links between the degree of plant sensitivity to O 3 and plasticity of stomatal properties in response to O 3 . Studies in open-top chambers (OTCs) and in continuously stirred tank reactors (CSTRs) established a clear relationship between plant developmental stages, degrees of O 3 sensitivity and g s : while 'S156' had higher g s rates than 'R123' earlier in development, similar differences between 'BBL 290' and 'BBL 274' were observed at later stages. G s rates on the abaxial leaf surfaces of 'S156' and 'BBL 290', accompanied by low leaf temperatures, were significantly higher than their O 3 -tolerant counterparts. Exposure to O 3 in CSTRs had greater and more consistent impacts on both stomatal densities and aperture sizes of O 3 -sensitive cultivars. Stomatal densities were highest on the abaxial leaf surfaces of 'S156' and 'BBL 290' at higher O 3 concentrations (60 ppb), but the largest aperture sizes were recorded on the adaxial leaf surfaces at moderate O 3 concentrations (30 ppb). Exposure to O 3 eliminated aperture size differences on the adaxial leaf surfaces between sensitive and tolerant cultivars. Regardless of sensitivity to O 3 and treatment regimes, the smallest aperture sizes and highest stomatal densities were found on the abaxial leaf surface. Our studies showed that O 3 has the potential to affect stomatal plasticity and confirmed the presence of different control mechanisms for stomatal development on each leaf surface. This appeared to be more evident in O 3 -sensitive cultivars. - O 3 has the potential to affect stomatal development and the presence of different control mechanisms on each leaf surface is confirmed

Updated stomatal flux and flux-effect models for wheat for quantifying effects of ozone on grain yield, grain mass and protein yield.

Science.gov (United States)

Grünhage, Ludger; Pleijel, Håkan; Mills, Gina; Bender, Jürgen; Danielsson, Helena; Lehmann, Yvonne; Castell, Jean-Francois; Bethenod, Olivier

2012-06-01

Field measurements and open-top chamber experiments using nine current European winter wheat cultivars provided a data set that was used to revise and improve the parameterisation of a stomatal conductance model for wheat, including a revised value for maximum stomatal conductance and new functions for phenology and soil moisture. For the calculation of stomatal conductance for ozone a diffusivity ratio between O(3) and H(2)O in air of 0.663 was applied, based on a critical review of the literature. By applying the improved parameterisation for stomatal conductance, new flux-effect relationships for grain yield, grain mass and protein yield were developed for use in ozone risk assessments including effects on food security. An example of application of the flux model at the local scale in Germany shows that negative effects of ozone on wheat grain yield were likely each year and on protein yield in most years since the mid 1980s. Copyright © 2012 Elsevier Ltd. All rights reserved.

Recombination-mediated genetic engineering of a bacterial artificial chromosome clone of modified vaccinia virus Ankara (MVA.

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Matthew G Cottingham

2008-02-01

Full Text Available The production, manipulation and rescue of a bacterial artificial chromosome clone of Vaccinia virus (VAC-BAC in order to expedite construction of expression vectors and mutagenesis of the genome has been described (Domi & Moss, 2002, PNAS99 12415-20. The genomic BAC clone was 'rescued' back to infectious virus using a Fowlpox virus helper to supply transcriptional machinery. We apply here a similar approach to the attenuated strain Modified Vaccinia virus Ankara (MVA, now widely used as a safe non-replicating recombinant vaccine vector in mammals, including humans. Four apparently full-length, rescuable clones were obtained, which had indistinguishable immunogenicity in mice. One clone was shotgun sequenced and found to be identical to the parent. We employed GalK recombination-mediated genetic engineering (recombineering of MVA-BAC to delete five selected viral genes. Deletion of C12L, A44L, A46R or B7R did not significantly affect CD8(+ T cell immunogenicity in BALB/c mice, but deletion of B15R enhanced specific CD8(+ T cell responses to one of two endogenous viral epitopes (from the E2 and F2 proteins, in accordance with published work (Staib et al., 2005, J. Gen. Virol.86, 1997-2006. In addition, we found a higher frequency of triple-positive IFN-gamma, TNF-alpha and IL-2 secreting E3-specific CD8+ T-cells 8 weeks after vaccination with MVA lacking B15R. Furthermore, a recombinant vaccine capable of inducing CD8(+ T cells against an epitope from Plasmodium berghei was created using GalK counterselection to insert an antigen expression cassette lacking a tandem marker gene into the traditional thymidine kinase locus of MVA-BAC. MVA continues to feature prominently in clinical trials of recombinant vaccines against diseases such as HIV-AIDS, malaria and tuberculosis. Here we demonstrate in proof-of-concept experiments that MVA-BAC recombineering is a viable route to more rapid and efficient generation of new candidate mutant and recombinant

Resource use and efficiency, and stomatal responses to environmental drivers of oak and pine species in an Atlantic Coastal Plain forest

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Heidi J Renninger

2015-05-01

Full Text Available Pine-oak ecosystems are globally distributed even though differences in anatomy and leaf habit between many co-occurring oaks and pines suggest different strategies for resource use, efficiency and stomatal behavior. The New Jersey Pinelands contain sandy soils with low water- and nutrient-holding capacity providing an opportunity to examine trade-offs in resource uptake and efficiency. Therefore, we compared resource use in terms of transpiration rates and leaf nitrogen content and resource-use efficiency including water-use efficiency (WUE via gas exchange and leaf carbon isotopes and photosynthetic nitrogen-use efficiency (PNUE between oaks (Quercus alba, Q. prinus, Q. velutina and pines (Pinus rigida, P. echinata. We also determined environmental drivers (vapor pressure deficit (VPD, soil moisture, solar radiation of canopy stomatal conductance (GS estimated via sap flow and stomatal sensitivity to light and soil moisture. Net assimilation rates were similar between genera, but oak leaves used about 10% more water and pine foliage contained about 20% more N per unit leaf area. Therefore, oaks exhibited greater PNUE while pines had higher WUE based on gas exchange, although WUE from carbon isotopes was not significantly different. For the environmental drivers of GS, oaks had about 10% lower stomatal sensitivity to VPD normalized by reference stomatal conductance compared with pines. Pines exhibited a significant positive relationship between shallow soil moisture and GS, but only GS in Q. velutina was positively related to soil moisture. In contrast, stomatal sensitivity to VPD was significantly related to solar radiation in all oak species but only pines at one site. Therefore, oaks rely more heavily on groundwater resources but have lower WUE, while pines have larger leaf areas and nitrogen acquisition but lower PNUE demonstrating a trade-off between using water and nitrogen efficiently in a resource-limited ecosystem.

Resource use and efficiency, and stomatal responses to environmental drivers of oak and pine species in an Atlantic Coastal Plain forest.

Science.gov (United States)

Renninger, Heidi J; Carlo, Nicholas J; Clark, Kenneth L; Schäfer, Karina V R

2015-01-01

Pine-oak ecosystems are globally distributed even though differences in anatomy and leaf habit between many co-occurring oaks and pines suggest different strategies for resource use, efficiency and stomatal behavior. The New Jersey Pinelands contain sandy soils with low water- and nutrient-holding capacity providing an opportunity to examine trade-offs in resource uptake and efficiency. Therefore, we compared resource use in terms of transpiration rates and leaf nitrogen content and resource-use efficiency including water-use efficiency (WUE) via gas exchange and leaf carbon isotopes and photosynthetic nitrogen-use efficiency (PNUE) between oaks (Quercus alba, Q. prinus, Q. velutina) and pines (Pinus rigida, P. echinata). We also determined environmental drivers [vapor pressure deficit (VPD), soil moisture, solar radiation] of canopy stomatal conductance (GS) estimated via sap flow and stomatal sensitivity to light and soil moisture. Net assimilation rates were similar between genera, but oak leaves used about 10% more water and pine foliage contained about 20% more N per unit leaf area. Therefore, oaks exhibited greater PNUE while pines had higher WUE based on gas exchange, although WUE from carbon isotopes was not significantly different. For the environmental drivers of GS, oaks had about 10% lower stomatal sensitivity to VPD normalized by reference stomatal conductance compared with pines. Pines exhibited a significant positive relationship between shallow soil moisture and GS, but only GS in Q. velutina was positively related to soil moisture. In contrast, stomatal sensitivity to VPD was significantly related to solar radiation in all oak species but only pines at one site. Therefore, oaks rely more heavily on groundwater resources but have lower WUE, while pines have larger leaf areas and nitrogen acquisition but lower PNUE demonstrating a trade-off between using water and nitrogen efficiently in a resource-limited ecosystem.

What determines the complex kinetics of stomatal conductance under blueless PAR in Festuca arundinacea? Subsequent effects on leaf transpiration.

Science.gov (United States)

Barillot, Romain; Frak, Ela; Combes, Didier; Durand, Jean-Louis; Escobar-Gutiérrez, Abraham J

2010-06-01

Light quality and, in particular, its content of blue light is involved in plant functioning and morphogenesis. Blue light variation frequently occurs within a stand as shaded zones are characterized by a simultaneous decrease of PAR and blue light levels which both affect plant functioning, for example, gas exchange. However, little is known about the effects of low blue light itself on gas exchange. The aims of the present study were (i) to characterize stomatal behaviour in Festuca arundinacea leaves through leaf gas exchange measurements in response to a sudden reduction in blue light, and (ii) to test the putative role of Ci on blue light gas exchange responses. An infrared gas analyser (IRGA) was used with light transmission filters to study stomatal conductance (gs), transpiration (Tr), assimilation (A), and intercellular concentration of CO(2) (Ci) responses to blueless PAR (1.80 mumol m(-2) s(-1)). The results were compared with those obtained under a neutral filter supplying a similar photosynthetic efficiency to the blueless PAR filter. It was shown that the reduction of blue light triggered a drastic and instantaneous decrease of gs by 43.2% and of Tr by 40.0%, but a gradual stomatal reopening began 20 min after the start of the low blue light treatment, thus leading to new steady-states. This new stomatal equilibrium was supposed to be related to Ci. The results were confirmed in more developed plants although they exhibited delayed and less marked responses. It is concluded that stomatal responses to blue light could play a key role in photomorphogenetic mechanisms through their effect on transpiration.

Contribution of competition for light to within-species variability in stomatal conductance

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Loranty, Michael M.; Mackay, D. Scott; Ewers, Brent E.; Traver, Elizabeth; Kruger, Eric L.

2010-05-01

Sap flux (JS) measurements were collected across two stands dominated by either trembling aspen or sugar maple in northern Wisconsin. Observed canopy transpiration (EC-obs) values derived from JS were used to parameterize the Terrestrial Regional Ecosystem Exchange Simulator ecosystem model. Modeled values of stomatal conductance (GS) were used to determine reference stomatal conductance (GSref), a proxy for GS that removes the effects of temporal responses to vapor pressure deficit (D) on spatial patterns of GS. Values of GSref were compared to observations of soil moisture, several physiological variables, and a competition index (CI) derived from a stand inventory, to determine the underlying cause of observed variability. Considerable variability in GSref between individual trees was found, with values ranging from 20 to 200 mmol m-2 s-1 and 20 to 100 mmol m-2 s-1 at the aspen and maple stands, respectively. Model-derived values of GSref and a sensitivity to D parameter (m) showed good agreement with a known empirical relationship for both stands. At both sites, GSref did not vary with topographic position, as indicated by surface soil moisture. No relationships were observed between GSref and tree height (HT), and a weak correlation with sapwood area (AS) was only significant for aspen. Significant nonlinear inverse relationships between GSref and CI were observed at both stands. Simulations with uniform reductions in incident photosynthetically active radiation (Q0) resulted in better agreement between observed and simulated EC. Our results suggest a link between photosynthesis and plant hydraulics whereby individual trees subject to photosynthetic limitation as a result of competitive shading exhibit a dynamic stomatal response resulting in a more conservative strategy for managing hydrologic resources.

Synthetically derived bat influenza A-like viruses reveal a cell type- but not species-specific tropism.

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Moreira, Étori Aguiar; Locher, Samira; Kolesnikova, Larissa; Bolte, Hardin; Aydillo, Teresa; García-Sastre, Adolfo; Schwemmle, Martin; Zimmer, Gert

2016-10-24

Two novel influenza A-like viral genome sequences have recently been identified in Central and South American fruit bats and provisionally designated "HL17NL10" and "HL18NL11." All efforts to isolate infectious virus from bats or to generate these viruses by reverse genetics have failed to date. Recombinant vesicular stomatitis virus (VSV) encoding the hemagglutinin-like envelope glycoproteins HL17 or HL18 in place of the VSV glycoprotein were generated to identify cell lines that are susceptible to bat influenza A-like virus entry. More than 30 cell lines derived from various species were screened but only a few cell lines were found to be susceptible, including Madin-Darby canine kidney type II (MDCK II) cells. The identification of cell lines susceptible to VSV chimeras allowed us to recover recombinant HL17NL10 and HL18NL11 viruses from synthetic DNA. Both influenza A-like viruses established a productive infection in MDCK II cells; however, HL18NL11 replicated more efficiently than HL17NL10 in this cell line. Unlike conventional influenza A viruses, bat influenza A-like viruses started the infection preferentially at the basolateral membrane of polarized MDCK II cells; however, similar to conventional influenza A viruses, bat influenza A-like viruses were released primarily from the apical site. The ability of HL18NL11 or HL17NL10 viruses to infect canine and human cells might reflect a zoonotic potential of these recently identified bat viruses.

Respiratory syncytial virus, pneumonia virus of mice, and influenza A virus differently affect respiratory allergy in mice

NARCIS (Netherlands)

Barends, M.; de Rond, L. G. H.; Dormans, J.; van Oosten, M.; Boelen, A.; Neijens, H. J.; Osterhaus, A. D. M. E.; Kimman, T. G.

2004-01-01

Respiratory viral infections in early childhood may interact with the immune system and modify allergen sensitization and/or allergic manifestations. In mice, respiratory syncytial virus (RSV) infection during allergic provocation aggravates the allergic T helper (Th) 2 immune response,

Antagonism of the Sodium-Potassium ATPase Impairs Chikungunya Virus Infection

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Alison W. Ashbrook

2016-05-01

Full Text Available Chikungunya virus (CHIKV is a reemerging alphavirus that has caused epidemics of fever, arthralgia, and rash worldwide. There are currently no licensed vaccines or antiviral therapies available for the prevention or treatment of CHIKV disease. We conducted a high-throughput, chemical compound screen that identified digoxin, a cardiac glycoside that blocks the sodium-potassium ATPase, as a potent inhibitor of CHIKV infection. Treatment of human cells with digoxin or a related cardiac glycoside, ouabain, resulted in a dose-dependent decrease in infection by CHIKV. Inhibition by digoxin was cell type-specific, as digoxin treatment of either murine or mosquito cells did not diminish CHIKV infection. Digoxin displayed antiviral activity against other alphaviruses, including Ross River virus and Sindbis virus, as well as mammalian reovirus and vesicular stomatitis virus. The digoxin-mediated block to CHIKV and reovirus infection occurred at one or more postentry steps, as digoxin inhibition was not bypassed by fusion of CHIKV at the plasma membrane or infection with cell surface-penetrating reovirus entry intermediates. Selection of digoxin-resistant CHIKV variants identified multiple mutations in the nonstructural proteins required for replication complex formation and synthesis of viral RNA. These data suggest a role for the sodium-potassium ATPase in promoting postentry steps of CHIKV replication and provide rationale for modulation of this pathway as a broad-spectrum antiviral strategy.

Structure of the Nucleoprotein Binding Domain of Mokola Virus Phosphoprotein▿

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Assenberg, René; Delmas, Olivier; Ren, Jingshan; Vidalain, Pierre-Olivier; Verma, Anil; Larrous, Florence; Graham, Stephen C.; Tangy, Frédéric; Grimes, Jonathan M.; Bourhy, Hervé

2010-01-01

Mokola virus (MOKV) is a nonsegmented, negative-sense RNA virus that belongs to the Lyssavirus genus and Rhabdoviridae family. MOKV phosphoprotein P is an essential component of the replication and transcription complex and acts as a cofactor for the viral RNA-dependent RNA polymerase. P recruits the viral polymerase to the nucleoprotein-bound viral RNA (N-RNA) via an interaction between its C-terminal domain and the N-RNA complex. Here we present a structure for this domain of MOKV P, obtained by expression of full-length P in Escherichia coli, which was subsequently truncated during crystallization. The structure has a high degree of homology with P of rabies virus, another member of Lyssavirus genus, and to a lesser degree with P of vesicular stomatitis virus (VSV), a member of the related Vesiculovirus genus. In addition, analysis of the crystal packing of this domain reveals a potential binding site for the nucleoprotein N. Using both site-directed mutagenesis and yeast two-hybrid experiments to measure P-N interaction, we have determined the relative roles of key amino acids involved in this interaction to map the region of P that binds N. This analysis also reveals a structural relationship between the N-RNA binding domain of the P proteins of the Rhabdoviridae and the Paramyxoviridae. PMID:19906936

Virus-Heat Shock Protein Interaction and a Novel Axis for Innate Antiviral Immunity

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Michael Oglesbee

2012-09-01

Full Text Available Virus infections induce heat shock proteins that in turn enhance virus gene expression, a phenomenon that is particularly well characterized for the major inducible 70 kDa heat shock protein (hsp70. However, hsp70 is also readily induced by fever, a phylogenetically conserved response to microbial infections, and when released from cells, hsp70 can stimulate innate immune responses through toll like receptors 2 and 4 (TLR2 and 4. This review examines how the virus-hsp70 relationship can lead to host protective innate antiviral immunity, and the importance of hsp70 dependent stimulation of virus gene expression in this host response. Beginning with the well-characterized measles virus-hsp70 relationship and the mouse model of neuronal infection in brain, we examine data indicating that the innate immune response is not driven by intracellular sensors of pathogen associated molecular patterns, but rather by extracellular ligands signaling through TLR2 and 4. Specifically, we address the relationship between virus gene expression, extracellular release of hsp70 (as a damage associated molecular pattern, and hsp70-mediated induction of antigen presentation and type 1 interferons in uninfected macrophages as a novel axis of antiviral immunity. New data are discussed that examines the more broad relevance of this protective mechanism using vesicular stomatitis virus, and a review of the literature is presented that supports the probable relevance to both RNA and DNA viruses and for infections both within and outside of the central nervous system.

The influence of virus infections on antioxidant levels in the genetically modified plum variety "Honeysweet" (Prunus domestica L.

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Jiri Sochor

2015-08-01

Full Text Available It is well-known that polyphenolic compounds are found abundantly in fruit, but various kinds of diseases  lower these levels. This work measures total polyphenolic content, antioxidant activity and the levels of specific important antioxidants in fruits of the genetically modified (GM plum variety HoneySweet, trees  which were previously inoculated with a range of different virus infections.  These were the Plum Pox virus (PPV, Prune Dwarf virus (PDV and Apple Chlorotic Leaf-Spot virus (ACLSV. Uninoculated trees were used as controls. Antioxidant activity was measured using four different photometric  methods – DPPH (2,2-diphenyl-1-picrylhydrazyl, DMPD (N-dimethyl-p-phenylenediamine, ABTS (2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid and FRAP (Ferric reducing antioxidant power. Total polyphenol content was measured using the Folin–Ciocalteau method. The profiles of 10 specific antioxidant constituents in the fruits of the GM plum variety HoneySweet were detected and analyzed, since these are of interest for their role in human diets and could play a role in the resistance of plants to viruses. Detection was made using HPLC with UV-VIS detection.  They were: gallic acid, p-coumaric acid, 4-aminobenzoic acid, chlorogenic acid, caffeic acid, ferulic acid, vanillin, rutin and quercetin. The compound with the highest concentration was chlorogenic acid (587 mg/100 g, and that with the lowest was p-coumaric acid (0.95 mg/100 g. Of the four methods of antioxidant activity used, in three the lowest levels of antioxidant activity were seen where the PPV virus was combined with ACLSV, and in three the highest levels were seen in the un-inoculated control without any infection. The highest values of total polyphenols were seen in the control (65.3 mg/100 g, followed by infection of PPV, then treatment PPV, PDV and ACLSV, then treatment PPV and PDV and finally the lowest levels were seen in treatment PPV and ACLSV (44.2 mg/100 g, which

Electrical signaling, stomatal conductance, ABA and Ethylene content in avocado trees in response to root hypoxia

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Gurovich, Luis; Schaffer, Bruce; García, Nicolás; Iturriaga, Rodrigo

2009-01-01

Avocado (Persea americana Mill.) trees are among the most sensitive of fruit tree species to root hypoxia as a result of flooded or poorly drained soil. Similar to drought stress, an early physiological response to root hypoxia in avocado is a reduction of stomatal conductance. It has been previously determined in avocado trees that an extracellular electrical signal between the base of stem and leaves is produced and related to reductions in stomatal conductance in response to drought stress. The current study was designed to determine if changes in the extracellular electrical potential between the base of the stem and leaves in avocado trees could also be detected in response to short-term (min) or long-term (days) root hypoxia, and if these signals could be related to stomatal conductance (gs), root and leaf ABA and ACC concentrations, ethylene emission from leaves and leaf abscission. In contrast to previous observations for drought-stressed trees, short-term or long-term root hypoxia did not stimulate an electrical potential difference between the base of the stem and leaves. Short-term hypoxia did not result in a significant decrease in gs compared with plants in the control treatment, and no differences in ABA concentration were found between plants subjected to hypoxia and control plants. Long-term hypoxia in the root zone resulted in a significant decrease in gs, increased leaf ethylene and increased leaf abscission. The results indicate that for avocado trees exposed to root hypoxia, electrical signals do not appear to be the primary root-to-shoot communication mechanism involved in signaling for stomatal closure as a result of hypoxia in the root zone. PMID:19649181

Membrane fusion-competent virus-like proteoliposomes and proteinaceous supported bilayers made directly from cell plasma membranes.

Science.gov (United States)

Costello, Deirdre A; Hsia, Chih-Yun; Millet, Jean K; Porri, Teresa; Daniel, Susan

2013-05-28

Virus-like particles are useful materials for studying virus-host interactions in a safe manner. However, the standard production of pseudovirus based on the vesicular stomatitis virus (VSV) backbone is an intricate procedure that requires trained laboratory personnel. In this work, a new strategy for creating virus-like proteoliposomes (VLPLs) and virus-like supported bilayers (VLSBs) is presented. This strategy uses a cell blebbing technique to induce the formation of nanoscale vesicles from the plasma membrane of BHK cells expressing the hemagglutinin (HA) fusion protein of influenza X-31. These vesicles and supported bilayers contain HA and are used to carry out single particle membrane fusion events, monitored using total internal reflection fluorescence microscopy. The results of these studies show that the VLPLs and VLSBs contain HA proteins that are fully competent to carry out membrane fusion, including the formation of a fusion pore and the release of fluorophores loaded into vesicles. This new strategy for creating spherical and planar geometry virus-like membranes has many potential applications. VLPLs could be used to study fusion proteins of virulent viruses in a safe manner, or they could be used as therapeutic delivery particles to transport beneficial proteins coexpressed in the cells to a target cell. VLSBs could facilitate high throughput screening of antiviral drugs or pathogen-host cell interactions.

Modeling daily gas exchange of a Douglas-fir forest : comparison of three stomatal conductance models with and without a soil water stress function

NARCIS (Netherlands)

Wijk, van M.T.; Dekker, S.C.; Bouten, W.; Bosveld, F.C.; Kohsiek, W.; Kramer, K.; Mohren, G.M.J.

2000-01-01

Modeling stomatal conductance is a key element in predicting tree growth and water use at the stand scale. We compared three commonly used models of stomatal conductance, the Jarvis-Loustau, Ball-Berry and Leuning models, for their suitability for incorporating soil water stress into their

Stomatal Conductance, Plant Hydraulics, and Multilayer Canopies: A New Paradigm for Earth System Models or Unnecessary Uncertainty

Science.gov (United States)

Bonan, G. B.

2016-12-01

Soil moisture stress is a key regulator of canopy transpiration, the surface energy budget, and land-atmosphere coupling. Many land surface models used in Earth system models have an ad-hoc parameterization of soil moisture stress that decreases stomatal conductance with soil drying. Parameterization of soil moisture stress from more fundamental principles of plant hydrodynamics is a key research frontier for land surface models. While the biophysical and physiological foundations of such parameterizations are well-known, their best implementation in land surface models is less clear. Land surface models utilize a big-leaf canopy parameterization (or two big-leaves to represent the sunlit and shaded canopy) without vertical gradients in the canopy. However, there are strong biometeorological and physiological gradients in plant canopies. Are these gradients necessary to resolve? Here, I describe a vertically-resolved, multilayer canopy model that calculates leaf temperature and energy fluxes, photosynthesis, stomatal conductance, and leaf water potential at each level in the canopy. In this model, midday leaf water stress manifests in the upper canopy layers, which receive high amounts of solar radiation, have high leaf nitrogen and photosynthetic capacity, and have high stomatal conductance and transpiration rates (in the absence of leaf water stress). Lower levels in the canopy become water stressed in response to longer-term soil moisture drying. I examine the role of vertical gradients in the canopy microclimate (solar radiation, air temperature, vapor pressure, wind speed), structure (leaf area density), and physiology (leaf nitrogen, photosynthetic capacity, stomatal conductance) in determining above canopy fluxes and gradients of transpiration and leaf water potential within the canopy.

Moderate restriction of macrophage-tropic human immunodeficiency virus type 1 by SAMHD1 in monocyte-derived macrophages.

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Taya, Kahoru; Nakayama, Emi E; Shioda, Tatsuo

2014-01-01

Macrophage-tropic human immunodeficiency virus type 1 (HIV-1) strains are able to grow to high titers in human monocyte-derived macrophages. However, it was recently reported that cellular protein SAMHD1 restricts HIV-1 replication in human cells of the myeloid lineage, including monocyte-derived macrophages. Here we show that degradation of SAMHD1 in monocyte-derived macrophages was associated with moderately enhanced growth of the macrophage-tropic HIV-1 strain. SAMHD1 degradation was induced by treating target macrophages with vesicular stomatitis virus glycoprotein-pseudotyped human immunodeficiency virus type 2 (HIV-2) particles containing viral protein X. For undifferentiated monocytes, HIV-2 particle treatment allowed undifferentiated monocytes to be fully permissive for productive infection by the macrophage-tropic HIV-1 strain. In contrast, untreated monocytes were totally resistant to HIV-1 replication. These results indicated that SAMHD1 moderately restricts even a macrophage-tropic HIV-1 strain in monocyte-derived macrophages, whereas the protein potently restricts HIV-1 replication in undifferentiated monocytes.

Global CO2 rise leads to reduced maximum stomatal conductance in Florida vegetation

NARCIS (Netherlands)

Lammertsma, E.I.; de Boer, H.J.; Dekker, S.C.; Dilcher, D.L.; Lotter, A.F.; Wagner-Cremer, F.

2011-01-01

A principle response of C3 plants to increasing concentrations of atmospheric CO2 (CO2) is to reduce transpirational water loss by decreasing stomatal conductance (gs) and simultaneously increase assimilation rates. Via this adaptation, vegetation has the ability to alter hydrology and climate.

Serologic survey for selected arboviruses and other potential pathogens in wildlife from Mexico.

Science.gov (United States)

Aguirre, A A; McLean, R G; Cook, R S; Quan, T J

1992-07-01

During 1988 and 1989, a serologic survey of wildlife was conducted in northeastern Mexico to determine the presence, prevalence, and distribution of arboviruses and other selected disease agents. Eighty mammal specimens were tested. Antibodies to vesicular stomatitis-Indiana, Venezuelan equine encephalitis-Mena II, Rio Grande virus, and vesicular stomatitis-New Jersey were detected predominantly in small mammals. Deer and mouflon (Ovis musimon) had antibodies to bluetongue and epizootic hemorrhagic disease. Two species had serologic evidence of recent exposure to Francisella tularensis. A white-tailed deer (Odocoileus virginianus) had antibodies to Anaplasma marginale. All specimens tested for antibodies against Yersinia pestis and Brucella abortus were negative. Sera from 315 birds were tested for antibody against five equine encephalitis viruses and six avian pathogens. During 1988, antibodies to Venezuelan equine encephalitis-Mena II, Venezuelan equine encephalitis-TC83, St. Louis encephalitis, eastern equine encephalitis, and western equine encephalitis were detected in birds of several species. Antibodies to Pasteurella multocida and Newcastle disease virus were also detected. Birds from five species presented antibodies to Mycoplasma meleagridis. Specimens tested for M. gallisepticum, M. synoviae, and Chlamydia psittaci were negative. To the best of our knowledge, this survey represents the first serologic evidence of bluetongue, Cache Valley virus, epizootic hemorrhagic disease, Jamestown Canyon virus, vesicular stomatitis-Indiana, vesicular stomatitis-New Jersey, Rio Grande virus, and tularemia reported among wildlife in Mexico.

Resistance of genetically modified potatoes to Potato virus Y under field conditions Resistência de plantas de batata geneticamente modificadas ao Potato virus Y em condições de campo

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André Nepomuceno Dusi

2009-09-01

Full Text Available The objective of this work was to evaluate the resistance of genetically modified clones of potato to Potato virus Y (PVY under field conditions. Genetically modified plants were compared with nontransformed plants of the same cultivar. The plots were flanked with potato plants infected with both PVYº and PVY N strains (spread lines, in order to provide the experimental area with the source of virus, which was naturally spread by the native aphid population. The experiment was weekly monitored by visual inspections and by DAS-Elisa in the plants produced from the harvested tubers, in order to evaluate the resistance of transgenic plants throughout the plant growth cycle. By the end of the third year, no infection symptoms were observed in the 1P clone; clone 63P showed 1% of infection, in contrast to about 90% of nontransformed plants infected. The stable expression of resistance to PVY provided by the coat protein gene was obtained in genetically modified clones of potato plants cultivar Achat under field conditions, during three consecutive years.O objetivo deste trabalho foi avaliar a resistência de clones geneticamente modificados de batata ao Potato virus Y (PVY em condições de campo. As plantas geneticamente modificadas foram comparadas com plantas não modificadas da mesma cultivar. As parcelas foram delimitadas com plantas infectadas com as estirpes PVYº e PVY N (linhas disseminadoras, para tornar disponível, na área experimental, a fonte de inóculo de vírus, que foi naturalmente disseminada pela população nativa de afídeos. O experimento foi monitorado semanalmente por inspeção visual e por DAS-Elisa nas plantas produzidas a partir dos tubérculos colhidos, para avaliar a resistência de plantas transgênicas ao longo do ciclo de crescimento. Ao final do terceiro ano, nenhum sintoma de infecção foi observado no clone 1P; o clone 63P apresentou 1% de infecção, em contraste com cerca de 90% de plantas-controle infectadas

Species climate range influences hydraulic and stomatal traits in Eucalyptus species.

Science.gov (United States)

Bourne, Aimee E; Creek, Danielle; Peters, Jennifer M R; Ellsworth, David S; Choat, Brendan

2017-07-01

Plant hydraulic traits influence the capacity of species to grow and survive in water-limited environments, but their comparative study at a common site has been limited. The primary aim of this study was to determine whether selective pressures on species originating in drought-prone environments constrain hydraulic traits among related species grown under common conditions. Leaf tissue water relations, xylem anatomy, stomatal behaviour and vulnerability to drought-induced embolism were measured on six Eucalyptus species growing in a common garden to determine whether these traits were related to current species climate range and to understand linkages between the traits. Hydraulically weighted xylem vessel diameter, leaf turgor loss point, the water potential at stomatal closure and vulnerability to drought-induced embolism were significantly ( P Eucalyptus trees has important implications for the limits of species responses to changing environmental conditions and thus for species survival and distribution into the future, and yields new information for physiological models. © The Author 2017. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For Permissions, please email: journals.permissions@oup.com

Three-Year Durability of Immune Responses Induced by HIV-DNA and HIV-Modified Vaccinia Virus Ankara and Effect of a Late HIV-Modified Vaccinia Virus Ankara Boost in Tanzanian Volunteers.

Science.gov (United States)

Joachim, Agricola; Munseri, Patricia J; Nilsson, Charlotta; Bakari, Muhammad; Aboud, Said; Lyamuya, Eligius F; Tecleab, Teghesti; Liakina, Valentina; Scarlatti, Gabriella; Robb, Merlin L; Earl, Patricia L; Moss, Bernard; Wahren, Britta; Mhalu, Fred; Ferrari, Guido; Sandstrom, Eric; Biberfeld, Gunnel

2017-08-01

We explored the duration of immune responses and the effect of a late third HIV-modified vaccinia virus Ankara (MVA) boost in HIV-DNA primed and HIV-MVA boosted Tanzanian volunteers. Twenty volunteers who had previously received three HIV-DNA and two HIV-MVA immunizations were given a third HIV-MVA immunization 3 years after the second HIV-MVA boost. At the time of the third HIV-MVA, 90% of the vaccinees had antibodies to HIV-1 subtype C gp140 (median titer 200) and 85% to subtype B gp160 (median titer 100). The majority of vaccinees had detectable antibody-dependent cellular cytotoxicity (ADCC)-mediating antibodies, 70% against CRF01_AE virus-infected cells (median titer 239) and 84% against CRF01_AE gp120-coated cells (median titer 499). A high proportion (74%) of vaccinees had IFN-γ ELISpot responses, 63% to Gag and 42% to Env, 3 years after the second HIV-MVA boost. After the third HIV-MVA, there was an increase in Env-binding antibodies and ADCC-mediating antibodies relative to the response seen at the time of the third HIV-MVA vaccination, p < .0001 and p < .05, respectively. The frequency of IFN-γ ELISpot responses increased to 95% against Gag or Env and 90% to both Gag and Env, p = .064 and p = .002, respectively. In conclusion, the HIV-DNA prime/HIV-MVA boost regimen elicited potent antibody and cellular immune responses with remarkable durability, and a third HIV-MVA immunization significantly boosted both antibody and cellular immune responses relative to the levels detected at the time of the third HIV-MVA, but not to higher levels than after the second HIV-MVA.

Rapid Titration of Measles and Other Viruses: Optimization with Determination of Replication Cycle Length

Science.gov (United States)

Grigorov, Boyan; Rabilloud, Jessica; Lawrence, Philip; Gerlier, Denis

2011-01-01

Background Measles virus (MV) is a member of the Paramyxoviridae family and an important human pathogen causing strong immunosuppression in affected individuals and a considerable number of deaths worldwide. Currently, measles is a re-emerging disease in developed countries. MV is usually quantified in infectious units as determined by limiting dilution and counting of plaque forming unit either directly (PFU method) or indirectly from random distribution in microwells (TCID50 method). Both methods are time-consuming (up to several days), cumbersome and, in the case of the PFU assay, possibly operator dependent. Methods/Findings A rapid, optimized, accurate, and reliable technique for titration of measles virus was developed based on the detection of virus infected cells by flow cytometry, single round of infection and titer calculation according to the Poisson's law. The kinetics follow up of the number of infected cells after infection with serial dilutions of a virus allowed estimation of the duration of the replication cycle, and consequently, the optimal infection time. The assay was set up to quantify measles virus, vesicular stomatitis virus (VSV), and human immunodeficiency virus type 1 (HIV-1) using antibody labeling of viral glycoprotein, virus encoded fluorescent reporter protein and an inducible fluorescent-reporter cell line, respectively. Conclusion Overall, performing the assay takes only 24–30 hours for MV strains, 12 hours for VSV, and 52 hours for HIV-1. The step-by-step procedure we have set up can be, in principle, applicable to accurately quantify any virus including lentiviral vectors, provided that a virus encoded gene product can be detected by flow cytometry. PMID:21915289

Acquired changes in stomatal characteristics in response to ozone during plant growth and leaf development of bush beans (Phaseolus vulgaris L.) indicate phenotypic plasticity

Energy Technology Data Exchange (ETDEWEB)

Elagoez, Vahram [Plant Biology Graduate Program, University of Massachusetts, Amherst, MA 01003 (United States)]. E-mail: velagoz@nsm.umass.edu; Han, Susan S. [Department of Plant, Soil and Insect Sciences, University of Massachusetts, Amherst, MA 01003 (United States); Manning, William J. [Department of Plant, Soil and Insect Sciences, University of Massachusetts, Amherst, MA 01003 (United States)

2006-04-15

Bush bean (Phaseolus vulgaris L.) lines 'S156' (O{sub 3}-sensitive)/'R123' (O{sub 3}-tolerant) and cultivars 'BBL 290' (O{sub 3}-sensitive)/'BBL 274' (O{sub 3}-tolerant) were used to study the effects of O{sub 3} on stomatal conductance (g {sub s}), density, and aperture size on leaf and pod surfaces with the objective of establishing links between the degree of plant sensitivity to O{sub 3} and plasticity of stomatal properties in response to O{sub 3}. Studies in open-top chambers (OTCs) and in continuously stirred tank reactors (CSTRs) established a clear relationship between plant developmental stages, degrees of O{sub 3} sensitivity and g {sub s}: while 'S156' had higher g {sub s} rates than 'R123' earlier in development, similar differences between 'BBL 290' and 'BBL 274' were observed at later stages. G {sub s} rates on the abaxial leaf surfaces of 'S156' and 'BBL 290', accompanied by low leaf temperatures, were significantly higher than their O{sub 3}-tolerant counterparts. Exposure to O{sub 3} in CSTRs had greater and more consistent impacts on both stomatal densities and aperture sizes of O{sub 3}-sensitive cultivars. Stomatal densities were highest on the abaxial leaf surfaces of 'S156' and 'BBL 290' at higher O{sub 3} concentrations (60 ppb), but the largest aperture sizes were recorded on the adaxial leaf surfaces at moderate O{sub 3} concentrations (30 ppb). Exposure to O{sub 3} eliminated aperture size differences on the adaxial leaf surfaces between sensitive and tolerant cultivars. Regardless of sensitivity to O{sub 3} and treatment regimes, the smallest aperture sizes and highest stomatal densities were found on the abaxial leaf surface. Our studies showed that O{sub 3} has the potential to affect stomatal plasticity and confirmed the presence of different control mechanisms for stomatal development on each leaf surface. This

Simulation of stomatal conductance for Aleppo pine to estimate its ozone uptake

Energy Technology Data Exchange (ETDEWEB)

Elvira, Susana [Ecotoxicology of Air Pollution, CIEMAT, Avda. Complutense 22 (ed. 70), 28040 Madrid (Spain); Alonso, Rocio [Ecotoxicology of Air Pollution, CIEMAT, Avda. Complutense 22 (ed. 70), 28040 Madrid (Spain); Gimeno, Benjamin S. [Ecotoxicology of Air Pollution, CIEMAT, Avda. Complutense 22 (ed. 70), 28040 Madrid (Spain)]. E-mail: benjamin.gimeno@ciemat.es

2007-04-15

The data from a previous experiment carried out in open-top chambers to assess the effects of ozone (O{sub 3}) exposure on growth and physiology of Aleppo pine (Pinus halepensis Mill.) were re-assessed to test the performance of the EMEP O{sub 3} stomatal conductance model used to estimate tree O{sub 3} uptake at a European scale. Aleppo pine seedlings were exposed during three consecutive years to three different O{sub 3} treatments: charcoal filtered air, non-filtered air and non-filtered air supplemented with 40 nl l{sup -1}. The results of the model using the default parameterisation already published for Mediterranean conifers showed a poor performance when compared to measured data. Therefore, modifications of g {sub max}, f {sub min}, and new f {sub VPD}, f {sub temp} and f {sub phen} functions were developed according to the observed data. This re-parameterisation resulted in a significant improvement of the performance of the model when compared to its original version. - Current EMEP stomatal uptake module needs to be re-parameterised for Mediterranean tree species.

Simulation of stomatal conductance for Aleppo pine to estimate its ozone uptake

International Nuclear Information System (INIS)

Elvira, Susana; Alonso, Rocio; Gimeno, Benjamin S.

2007-01-01

The data from a previous experiment carried out in open-top chambers to assess the effects of ozone (O 3 ) exposure on growth and physiology of Aleppo pine (Pinus halepensis Mill.) were re-assessed to test the performance of the EMEP O 3 stomatal conductance model used to estimate tree O 3 uptake at a European scale. Aleppo pine seedlings were exposed during three consecutive years to three different O 3 treatments: charcoal filtered air, non-filtered air and non-filtered air supplemented with 40 nl l -1 . The results of the model using the default parameterisation already published for Mediterranean conifers showed a poor performance when compared to measured data. Therefore, modifications of g max , f min , and new f VPD , f temp and f phen functions were developed according to the observed data. This re-parameterisation resulted in a significant improvement of the performance of the model when compared to its original version. - Current EMEP stomatal uptake module needs to be re-parameterised for Mediterranean tree species

Stomatal vs. genome size in angiosperms: the somatic tail wagging the genomic dog?

NARCIS (Netherlands)

Hodgson, J.G.; Sharafi, M.; Jalili, A.; Diaz, S.; Montserrat-Marti, G.; Palmer, C.; Cerabolini, B.; Pierce, S.; Hamzehee, B.; Asri, Y.; Jamzad, Z.; Wilson, P.; Zarrinkamar, F.; Raven, J.; Band, S.R.; Basconcelo, S.; Bogard, A.; Carter, G.; Charles, M.; Castro-Diez, P.; Cornelissen, J.H.C.; Funes, G.; Jones, M.; Khoshnevis, M.; Perez-Harguindeguy, N.; Perez-Rontome, M.C.; Shirvany, F.A.; Vendramini, F.; Yazdani, S.; Abbas-Azimi, R.; Boustani, S.; Dehghan, M.; Hynd, F.A.; Kowsary, E.; Kazemi-Saeed, F.; Siavash, B.; Villar-Salvador, P.; Cragie, R.; Naqinezhad, A.; Romo-Diez, A.; De Torres Espuny, L.; Simmons, E.

2010-01-01

Background and Aims Genome size is a function, and the product, of cell volume. As such it is contingent on ecological circumstance. The nature of 'this ecological circumstance' is, however, hotly debated. Here, we investigate for angiosperms whether stomatal size may be this 'missing link': the

Canine Parvovirus (CPV) Vaccination: Comparison of Neutralizing Antibody Responses in Pups after Inoculation with CPV2 or CPV2b Modified Live Virus Vaccine

Science.gov (United States)

Pratelli, Annamaria; Cavalli, Alessandra; Martella, Vito; Tempesta, Maria; Decaro, Nicola; Carmichael, Leland Eugene; Buonavoglia, Canio

2001-01-01

Canine parvovirus type 2 (CPV2) emerged in 1978 as causative agent of a new disease of dogs. New antigenic variants (biotypes), designated CPV2a and CPV2b, became widespread during 1979 to 1980 and 1984, respectively. At the present time the original CPV2 has disappeared in the dog population and has been replaced by the two new viruses. In the present study the comparison of neutralizing antibody titers in two groups of pups (18 pups in each group) inoculated with CPV2 and CPV2b modified live virus vaccines is reported. Using the hemagglutination inhibition (HI) test, relevant differences between antibody titers, against either the homologous or the heterologous virus, were not constantly observed. Using the neutralization (Nt) test, however, the pups inoculated with CPV2 had antibody titers which were approximately 30 times higher to the homologous virus (mean, 4,732) than to the heterologous virus (CPV2b) (mean, 162). The results of these experiments support two conclusions: (i) the HI test may not always accurately evaluate the true immune status of dogs with respect to CPV, and (ii) dogs inoculated with CPV2 vaccine develop relatively low Nt antibody titers against the heterologous virus (CPV2b). These data may suggest an advantage for new vaccines, considering that most presently licensed vaccines are produced with CPV2, which no longer exists in the dog population. PMID:11329467

Canine parvovirus (CPV) vaccination: comparison of neutralizing antibody responses in pups after inoculation with CPV2 or CPV2b modified live virus vaccine.

Science.gov (United States)

Pratelli, A; Cavalli, A; Martella, V; Tempesta, M; Decaro, N; Carmichael, L E; Buonavoglia, C

2001-05-01

Canine parvovirus type 2 (CPV2) emerged in 1978 as causative agent of a new disease of dogs. New antigenic variants (biotypes), designated CPV2a and CPV2b, became widespread during 1979 to 1980 and 1984, respectively. At the present time the original CPV2 has disappeared in the dog population and has been replaced by the two new viruses. In the present study the comparison of neutralizing antibody titers in two groups of pups (18 pups in each group) inoculated with CPV2 and CPV2b modified live virus vaccines is reported. Using the hemagglutination inhibition (HI) test, relevant differences between antibody titers, against either the homologous or the heterologous virus, were not constantly observed. Using the neutralization (Nt) test, however, the pups inoculated with CPV2 had antibody titers which were approximately 30 times higher to the homologous virus (mean, 4,732) than to the heterologous virus (CPV2b) (mean, 162). The results of these experiments support two conclusions: (i) the HI test may not always accurately evaluate the true immune status of dogs with respect to CPV, and (ii) dogs inoculated with CPV2 vaccine develop relatively low Nt antibody titers against the heterologous virus (CPV2b). These data may suggest an advantage for new vaccines, considering that most presently licensed vaccines are produced with CPV2, which no longer exists in the dog population.

Disruption of stomatal lineage signaling or transcriptional regulators has differential effects on mesophyll development, but maintains coordination of gas exchange.

Science.gov (United States)

Dow, Graham J; Berry, Joseph A; Bergmann, Dominique C

2017-10-01

Stomata are simultaneously tasked with permitting the uptake of carbon dioxide for photosynthesis while limiting water loss from the plant. This process is mainly regulated by guard cell control of the stomatal aperture, but recent advancements have highlighted the importance of several genes that control stomatal development. Using targeted genetic manipulations of the stomatal lineage and a combination of gas exchange and microscopy techniques, we show that changes in stomatal development of the epidermal layer lead to coupled changes in the underlying mesophyll tissues. This coordinated response tends to match leaf photosynthetic potential (V cmax ) with gas-exchange capacity (g smax ), and hence the uptake of carbon dioxide for water lost. We found that different genetic regulators systematically altered tissue coordination in separate ways: the transcription factor SPEECHLESS (SPCH) primarily affected leaf size and thickness, whereas peptides in the EPIDERMAL PATTERNING FACTOR (EPF) family altered cell density in the mesophyll. It was also determined that interlayer coordination required the cell-surface receptor TOO MANY MOUTHS (TMM). These results demonstrate that stomata-specific regulators can alter mesophyll properties, which provides insight into how molecular pathways can organize leaf tissues to coordinate gas exchange and suggests new strategies for improving plant water-use efficiency. © 2017 The Authors. New Phytologist © 2017 New Phytologist Trust.

The seroprevalence and salivary shedding of herpesviruses in Behçet's syndrome and recurrent aphthous stomatitis

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Noha Seoudi

2015-06-01

Full Text Available Background: Behçet's syndrome (BS is one of the multisystemic diseases that presents with oral ulceration and several other systemic manifestations including genital ulceration, folliculitis, erythema nodosum-like lesions, uveitis, and arthropathy. Ocular manifestation, central nervous system involvement, and gastrointestinal manifestation account for most of the complications of this disease, whereas orogenital ulceration and dermatological involvement affects the quality of life. The cause of the disease is not fully elucidated; however, herpesviruses have long been thought to play a pivotal role in the disease pathogenesis. Objective: To investigate the seroprevalence and salivary shedding of herpesviruses in BS. Method: The levels of specific immunoglobulin G in six different herpesviruses in serum samples collected from 54 BS, 28 healthy controls (HC, and 7 recurrent aphthous stomatitis (RAS patients were investigated. Salivary viral load was also quantified for these viruses in matched saliva samples using quantitative real-time polymerase chain reaction. Results: The BS had lower cytomegalovirus (CMV IgG level in comparison to HC (p=0.0226 and RAS (p=0.0450. There was statistically significant higher salivary shedding of Epstein-Barr virus (EBV in BS in comparison to HC (p=0.0052, but not RAS (p=0.3318. Conclusions: A high EBV shedding was observed in both BS and RAS and a lower level of CMV IgG was observed in BS only. The reason for the observed lower level of CMV IgG in BS is not clear. However, one explanation might be a defect in the cross-talk between innate and adaptive immune responses which was suggested by a previously described defect in the toll-like receptor 1 and 2 heterodimer formation and function, this being the initial receptor sensing of CMV.

Haiku: New paradigm for the reverse genetics of emerging RNA viruses.

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Thérèse Atieh

Full Text Available Reverse genetics is key technology for producing wild-type and genetically modified viruses. The ISA (Infectious Subgenomic Amplicons method is a recent versatile and user-friendly reverse genetics method to rescue RNA viruses. The main constraint of its canonic protocol was the requirement to produce (e.g., by DNA synthesis or fusion PCR 5' and 3' modified genomic fragments encompassing the human cytomegalovirus promoter (pCMV and the hepatitis delta virus ribozyme/simian virus 40 polyadenylation signal (HDR/SV40pA, respectively. Here, we propose the ultimately simplified "Haiku" designs in which terminal pCMV and HDR/SV40pA sequences are provided as additional separate DNA amplicons. This improved procedure was successfully applied to the rescue of a wide range of viruses belonging to genera Flavivirus, Alphavirus and Enterovirus in mosquito or mammalian cells using only standard PCR amplification techniques and starting from a variety of original materials including viral RNAs extracted from cell supernatant media or animal samples. We also demonstrate that, in specific experimental conditions, the presence of the HDR/SV40pA is not necessary to rescue the targeted viruses. These ultimately simplified "Haiku" designs provide an even more simple, rapid, versatile and cost-effective tool to rescue RNA viruses since only generation of overlapping amplicons encompassing the entire viral genome is now required to generate infectious virus. This new approach may completely modify our capacity to obtain infectious RNA viruses.

Utility of two modified-live virus canine distemper vaccines in wild-caught fishers (Martes pennanti).

Science.gov (United States)

Peper, Steven T; Peper, Randall L; Mitcheltree, Denise H; Kollias, George V; Brooks, Robert P; Stevens, Sadie S; Serfass, Thomas L

2016-12-01

Canine distemper virus (CDV) infects families in the order Carnivora. As a preventive measure, vaccinations against CDV are frequently given to mustelids in captive environments. Our objectives were to compare the utility between two modified-live virus canine distemper vaccines (MLV CDV's), Fervac-D® (no longer manufactured) and Galaxy-D® (now manufactured by MSD Animal Health as part of a multivalent vaccine), in developing an immune response in wild-caught fishers. The Pennsylvania Fisher Reintroduction Project (PFRP) used 14 wild-caught fishers during one year of the project to evaluate the utility of vaccinations against CDV as part of any reintroduction project. Fishers were injected subcutaneously in the nape of the neck with their designated vaccine. Fervac-D® did not effectively stimulate development of a serologic antibody response, whereas Galaxy-D® had adequate seroconversion or rise of titer levels to suggest that the general use of MLV CDV may be suitable in fishers pending further studies. We recommend that future studies be conducted, evaluating the use of currently produced vaccines in fishers. Future research should also focus on the length of days required between administration of primary and booster vaccines to achieve sufficient immune response. If only primary doses are required, then hard-release reintroduction projects for fishers could be recommended. If primary and booster vaccines are required then soft-release reintroduction projects should be recommended that include captive management periods, allowing for appropriate vaccination intervals needed to maximize the probability of protection against CDV.

Mesoporous Silicon with Modified Surface for Plant Viruses and Their Protein Particle Sensing

Directory of Open Access Journals (Sweden)

Kae Dal Kwack

2008-10-01

Full Text Available Changes in electric parameters of a mesoporous silicon treated by a plasma chemical etching with fluorine and hydrogen ions, under the adsorption of NEPO (Nematodetransmitted Polyhedral plant viruses such as TORSV (Tomato Ringspot Virus, GFLV (Grapevine Fan Leaf Virus and protein macromolecule from TORSV particles are described. The current response to the applied voltage is measured for each virus particle to investigate the material parameters which are sensitive to the adsorbed particles. The peculiar behaviors of the response are modeled by the current-voltage relationship in a MOSFET. This model explains the behavior well and the double gate model of the MOSFET informs that the mesoporous silicon is a highly sensitive means of detecting the viruses in the size range less than 50 nm.

Compromised Photosynthetic Electron Flow And H2O2 Generation Correlate with Genotype-Specific Stomatal Dysfunctions During Resistance Against Powdery Mildew In Oats.

Directory of Open Access Journals (Sweden)

Javier Sánchez-Martín

2016-11-01

Full Text Available Stomatal dysfunction known as locking has been linked to the elicitation of a hypersensitive response (HR following attack of fungal pathogens in cereals. We here assess how spatial and temporal patterns of different resistance mechanisms, such as HR and penetration resistance influence stomatal and photosynthetic parameters in oat (Avena sativa and the possible involvement of hydrogen peroxide (H2O2 in the dysfunctions observed. Four oat cultivars with differential resistance responses (i.e. penetration resistance, early and late HR to powdery mildew (Blumeria graminis f. sp. avenae, Bga were used. Results demonstrated that stomatal dysfunctions were genotype but not response-type dependent since genotypes with similar resistance responses when assessed histologically showed very different locking patterns. Maximum quantum yield (Fv/Fm of photosystem II were compromised in most Bga–oat interactions and photoinhibition increased. However, the extent of the photosynthetic alterations was not directly related to the extent of HR. H2O2 generation is triggered during the execution of resistance responses and can influence stomatal function. Artificially increasing H2O2 by exposing plants to increased light intensity further reduced Fv/Fm ratios and augmented the patterns of stomatal dysfunctions previously observed. The latter results suggest that the observed dysfunctions and hence a cost of resistance may be linked with oxidative stress occurring during defence induced photosynthetic disruption.

Plasticity in stomatal size and density of potato leaves under different irrigation and phosphorus regimes.

Science.gov (United States)

Sun, Yanqi; Yan, Fei; Cui, Xiaoyong; Liu, Fulai

2014-09-01

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 unresponsive to P but significantly affected by the irrigation treatment. FI plants had the largest SS, followed by DI, and PRD the smallest; and the reverse was the case for SD. Compared to FI and DI, PRD plants had significantly lower values of specific leaf area (SLA) and leaf carbon isotope discrimination (Δ(13)C) under P0. Midday leaf water potential (Ψleaf) and stomatal conductance (gs) was similar for DI and PRD, which was significantly lower than that of FI. Leaf contents of C, N, K, Ca and Mg were higher in PRD than in DI plants, particularly under P0. When analyzed across the three irrigation regimes, it was found that the P1 plants had significantly higher leaf contents of P and Mg, but significantly lower leaf K content compared to the P0 plants. Linear correlation analyses revealed that SS was positively correlated with Ψleaf and Δ(13)C; whereas SD was negatively correlated with Ψleaf, Δ(13)C and SLA, and positively correlated with leaf C, N and Ca contents. And gs was positively correlated with SS but negatively correlated with SD. Collectively, under low P level, the smaller and denser stomata in PRD plants may bring about a more efficient stomatal control over gas exchange, hereby potentially enhance water-use efficiency as exemplified by the lowered leaf Δ(13)C under fluctuating soil moisture conditions. Copyright © 2014 Elsevier GmbH. All rights reserved.

Analysis of the solvent accessibility of cysteine residues on Maize rayado fino virus virus-like particles produced in Nicotiana benthamiana plants and cross-linking of peptides to VLPs.

Science.gov (United States)

Natilla, Angela; Hammond, Rosemarie W

2013-02-14

Mimicking and exploiting virus properties and physicochemical and physical characteristics holds promise to provide solutions to some of the world's most pressing challenges. The sheer range and types of viruses coupled with their intriguing properties potentially give endless opportunities for applications in virus-based technologies. Viruses have the ability to self- assemble into particles with discrete shape and size, specificity of symmetry, polyvalence, and stable properties under a wide range of temperature and pH conditions. Not surprisingly, with such a remarkable range of properties, viruses are proposed for use in biomaterials, vaccines, electronic materials, chemical tools, and molecular electronic containers. In order to utilize viruses in nanotechnology, they must be modified from their natural forms to impart new functions. This challenging process can be performed through several mechanisms including genetic modification of the viral genome and chemically attaching foreign or desired molecules to the virus particle reactive groups. The ability to modify a virus primarily depends upon the physiochemical and physical properties of the virus. In addition, the genetic or physiochemical modifications need to be performed without adversely affecting the virus native structure and virus function. Maize rayado fino virus (MRFV) coat proteins self-assemble in Escherichia coli producing stable and empty VLPs that are stabilized by protein-protein interactions and that can be used in virus-based technologies applications. VLPs produced in tobacco plants were examined as a scaffold on which a variety of peptides can be covalently displayed. Here, we describe the steps to 1) determine which of the solvent-accessible cysteines in a virus capsid are available for modification, and 2) bioconjugate peptides to the modified capsids. By using native or mutationally-inserted amino acid residues and standard coupling technologies, a wide variety of materials have been

Antimicrobial photodynamic therapy for infectious stomatitis in snakes: Clinical views and microbiological findings.

Science.gov (United States)

Grego, Kathleen Fernandes; Carvalho, Marcelo Pires Nogueira de; Cunha, Marcos Paulo Vieira; Knöbl, Terezinha; Pogliani, Fabio Celidonio; Catão-Dias, José Luiz; Sant'Anna, Sávio Stefanini; Ribeiro, Martha Simões; Sellera, Fábio Parra

2017-12-01

Antimicrobial photodynamic therapy (APDT) has been broadly investigated as an alternative to treat localized infections, without leading to the selection of resistant microorganisms. Infectious stomatitis is a multifactorial disease frequently reported in captive snakes characterized by infection of the oral mucosa and surrounding tissues. In this study, we investigated methylene blue (MB)-mediated APDT to treat infectious stomatitis in snakes and verified the resistance phenotype and genotype before and after APDT. Three Boid snakes presented petechiae, edema and caseous material in their oral cavities. MB (0.01%) was applied on the lesions and after 5min they were irradiated using a red laser (λ=660nm), fluence of 280J/cm 2 , 8J and 80s per point, 100mW, spot size 0.028cm 2 and fluence rate of 3.5W/cm 2 . APDT was repeated once a week during 3 months. Samples of the lesions were collected to identify bacteria and antibiotic resistance profiles. To analyze the clonality of bacterial isolates before and after APDT, isolates were subjected to ERIC PCR analysis. Snakes presented clinical improvement such as reduction of inflammatory signs and caseous material. Pseudomonas aeruginosa and Escherichia coli were present in all snakes; Klebsiella pneumoniae and Morganella morganii were also identified in some animals. We also observed that the oral microbiota was completely replaced following APDT. However, K. pneumoniae isolates before and after APDT were a single clone with 100% of genetic similarity that lost resistance phenotype for seven antibiotics of four classes. These results show that APDT can be used to treat infectious stomatitis in snakes. Copyright © 2017 Elsevier B.V. All rights reserved.

Utilization of Dexaltin ointment to radiation stomatitis in patients with cancer of the oral cavity

International Nuclear Information System (INIS)

Ohhashi, Yasushi; Abe, Masaki; Ueda, Noboru

1981-01-01

Dexaltin ointment (sodium polyactylic acid, containing 0.1% dexamethasone) was used to the patients with stomatitis induced by radiotherapy. Fifteen patients with cancer of the oral cavity, aged from 44 to 77, were exposed to 60 Co γ-rays or electron beams, with the irradiation dose of 2400 to 9000 rad. About 60 g of the ointment was pasted for mean period of 6.6 weeks after meals, before sleep, and whenever the patients wanted. Therapeutic effect was observed in every case and in 87% of the case the pain at meals was decreased. The ointment adhered mostly for 1 - 2 hrs and in some cases, more than 3 hrs. No adverse reaction was observed. Therefore it was suggested that Dexaltin was a promising ointment to the radiation stomatitis. (Nakanishi, T.)

Testing of models of stomatal ozone fluxes with field measurements in a mixed Mediterranean forest

Czech Academy of Sciences Publication Activity Database

Fares, S.; Matteucci, G.; Mugnozza, S.; Morani, A.; Calfapietra, Carlo; Salvatori, E.; Fusaro, L.; Manes, F.; Loreto, F.

2013-01-01

Roč. 67, MAR (2013), s. 242-251 ISSN 1352-2310 Institutional support: RVO:67179843 Keywords : Ozone fluxes * Stomatal conductance models * GPP * Mediterranean forest Subject RIV: EH - Ecology, Behaviour Impact factor: 3.062, year: 2013

Improved survival in rhesus macaques immunized with modified vaccinia virus Ankara recombinants expressing simian immunodeficiency virus envelope correlates with reduction in memory CD4+ T-cell loss and higher titers of neutralizing antibody.

Science.gov (United States)

Ourmanov, Ilnour; Kuwata, Takeo; Goeken, Robert; Goldstein, Simoy; Iyengar, Ranjani; Buckler-White, Alicia; Lafont, Bernard; Hirsch, Vanessa M

2009-06-01

Previous studies demonstrated that immunization of macaques with simian immunodeficiency virus (SIV) Gag-Pol and Env recombinants of the attenuated poxvirus modified vaccinia virus Ankara (MVA) provided protection from high viremia and AIDS following challenge with a pathogenic strain of SIV. Although all animals became infected, plasma viremia was significantly reduced in animals that received the MVA-SIV recombinant vaccines compared with animals that received nonrecombinant MVA. Most importantly, the reduction in viremia resulted in a significant increase in median and cumulative survival. Continued analysis of these animals over the subsequent 9 years has shown that they maintain a survival advantage, although all but two of the macaques have progressed to AIDS. Importantly, improved survival correlated with preservation of memory CD4(+) T cells in the peripheral blood. The greatest survival advantage was observed in macaques immunized with regimens containing SIV Env, and the titer of neutralizing antibodies to the challenge virus prior to or shortly following challenge correlated with preservation of CD4(+) T cells. These data are consistent with a role for neutralizing antibodies in nonsterilizing protection from high viremia and associated memory CD4(+) T-cell loss.

Inhibition of Lassa virus glycoprotein cleavage and multicycle replication by site 1 protease-adapted alpha(1-antitrypsin variants.

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Anna Maisa

2009-06-01

Full Text Available Proteolytic processing of the Lassa virus envelope glycoprotein precursor GP-C by the host proprotein convertase site 1 protease (S1P is a prerequisite for the incorporation of the subunits GP-1 and GP-2 into viral particles and, hence, essential for infectivity and virus spread. Therefore, we tested in this study the concept of using S1P as a target to block efficient virus replication.We demonstrate that stable cell lines inducibly expressing S1P-adapted alpha(1-antitrypsin variants inhibit the proteolytic maturation of GP-C. Introduction of the S1P recognition motifs RRIL and RRLL into the reactive center loop of alpha(1-antitrypsin resulted in abrogation of GP-C processing by endogenous S1P to a similar level observed in S1P-deficient cells. Moreover, S1P-specific alpha(1-antitrypsins significantly inhibited replication and spread of a replication-competent recombinant vesicular stomatitis virus expressing the Lassa virus glycoprotein GP as well as authentic Lassa virus. Inhibition of viral replication correlated with the ability of the different alpha(1-antitrypsin variants to inhibit the processing of the Lassa virus glycoprotein precursor.Our data suggest that glycoprotein cleavage by S1P is a promising target for the development of novel anti-arenaviral strategies.

Method of inhibiting plant virus pathogen infections by crispr/cas9-mediated interference

KAUST Repository

Mahfouz, Magdy Mahmoud

2016-11-24

A genetically modified tobacco plant or tomato plant resistant to at least one pathogenic geminiviridae virus species is provided. The plant comprises a heterologous CRISPR/Cas9 system and at least one heterologous nucleotide sequence that is capable of hybridizing to a nucleotide sequence of the pathogenic virus and that directs inactivation of the pathogenic virus species or plurality of viral species by the CRISPR/Cas9 system. The heterologous nucleotide sequence can be complementary to, but not limited to an Intergenic Region (IR) of the Tomato Yellow Leaf Curl Virus (TYLCV), Further provided are methods of generating a genetically modified plant that is resistant to a virus pathogen by a heterologous CRISPR/Cas9 system and expression of a gRNA specifically targeting the virus.

CDPKs CPK6 and CPK3 function in ABA regulation of guard cell S-type anion- and Ca(2+-permeable channels and stomatal closure.

Directory of Open Access Journals (Sweden)

Izumi C Mori

2006-10-01

Full Text Available Abscisic acid (ABA signal transduction has been proposed to utilize cytosolic Ca(2+ in guard cell ion channel regulation. However, genetic mutants in Ca(2+ sensors that impair guard cell or plant ion channel signaling responses have not been identified, and whether Ca(2+-independent ABA signaling mechanisms suffice for a full response remains unclear. Calcium-dependent protein kinases (CDPKs have been proposed to contribute to central signal transduction responses in plants. However, no Arabidopsis CDPK gene disruption mutant phenotype has been reported to date, likely due to overlapping redundancies in CDPKs. Two Arabidopsis guard cell-expressed CDPK genes, CPK3 and CPK6, showed gene disruption phenotypes. ABA and Ca(2+ activation of slow-type anion channels and, interestingly, ABA activation of plasma membrane Ca(2+-permeable channels were impaired in independent alleles of single and double cpk3cpk6 mutant guard cells. Furthermore, ABA- and Ca(2+-induced stomatal closing were partially impaired in these cpk3cpk6 mutant alleles. However, rapid-type anion channel current activity was not affected, consistent with the partial stomatal closing response in double mutants via a proposed branched signaling network. Imposed Ca(2+ oscillation experiments revealed that Ca(2+-reactive stomatal closure was reduced in CDPK double mutant plants. However, long-lasting Ca(2+-programmed stomatal closure was not impaired, providing genetic evidence for a functional separation of these two modes of Ca(2+-induced stomatal closing. Our findings show important functions of the CPK6 and CPK3 CDPKs in guard cell ion channel regulation and provide genetic evidence for calcium sensors that transduce stomatal ABA signaling.

Leaf temperature and stomatal influences on sap velocity diurnal hysteresis in the Amazon rainforest

Science.gov (United States)

Jardine, K.; Gimenez, B.; Negron Juarez, R. I.; Koven, C.; Powell, T.; Higuchi, N.; Chambers, J.; Varadharajan, C.

2016-12-01

In order to improve our ability to predict terrestrial evapotranspiration fluxes, an understanding of the interactions between plant physiology and environmental conditions is necessary, but remains poorly characterized, especially in tropical ecosystems. In this study we show a tight positive correlation between sap velocity (at 1 m of height) and leaf surface temperature (LST, 20-30 m of height) in canopy dominant trees in two primary rainforest sites in the Amazon basin (Santarém and Manaus, Brazil). As leaf temperatures varied throughout the day, sap velocity responded with little delay (<15 min). Positive sap velocity was often observed at night, but also closely followed night time LSTs. When plotted versus LST, sap velocity showed an exponential increase before reaching a reflection point and a plateau and is characterized as a sigmoidal curve, in all observed trees. Moreover, a clear diurnal hysteresis in sap velocity was evident with morning periods showing higher temperature sensitivities than afternoon and night periods. Diurnal leaf observations showed a morning peak in stomatal conductance ( 10:00-10:30), but a mid-day to afternoon peak in transpiration and leaf temperature (12:00-14:00). Our observations suggest the sap velocity-LST hysteresis pattern arises due to the temporal offset between stomatal conductance and vapor pressure deficits (VPD) and demonstrates the dominating effect of VPD over stomatal conductance in maintaining high transpiration/sap flow rates under elevated temperatures. Our results have important implications for modeling tropical forest transpiration and suggests the possibility of predicting evapotranspiration fluxes at the ecosystem to regional scales based on remote sensed vegetation temperature.

A banana NAC transcription factor (MusaSNAC1) impart drought tolerance by modulating stomatal closure and H2O2 content.

Science.gov (United States)

Negi, Sanjana; Tak, Himanshu; Ganapathi, T R

2018-03-01

MusaSNAC1 function in H 2 O 2 mediated stomatal closure and promote drought tolerance by directly binding to CGT[A/G] motif in regulatory region of multiple stress-related genes. Drought is a abiotic stress-condition, causing reduced plant growth and diminished crop yield. Guard cells of the stomata control photosynthesis and transpiration by regulating CO 2 exchange and water loss, thus affecting growth and crop yield. Roles of NAC (NAM, ATAF1/2 and CUC2) protein in regulation of stress-conditions has been well documented however, their control over stomatal aperture is largely unknown. In this study we report a banana NAC protein, MusaSNAC1 which induced stomatal closure by elevating H 2 O 2 content in guard cells during drought stress. Overexpression of MusaSNAC1 in banana resulted in higher number of stomata closure causing reduced water loss and thus elevated drought-tolerance. During drought, expression of GUS (β-glucuronidase) under P MusaSNAC1 was remarkably elevated in guard cells of stomata which correlated with its function as a transcription factor regulating stomatal aperture closing. MusaSNAC1 is a transcriptional activator belonging to SNAC subgroup and its 5'-upstream region contain multiple Dof1 elements as well as stress-associated cis-elements. Moreover, MusaSNAC1 also regulate multiple stress-related genes by binding to core site of NAC-proteins CGT[A/G] in their 5'-upstream region. Results indicated an interesting mechanism of drought tolerance through stomatal closure by H 2 O 2 generation in guard cells, regulated by a NAC-protein in banana.

A combination in-ovo vaccine for avian influenza virus and Newcastle disease virus.

Science.gov (United States)

Steel, John; Burmakina, Svetlana V; Thomas, Colleen; Spackman, Erica; García-Sastre, Adolfo; Swayne, David E; Palese, Peter

2008-01-24

The protection of poultry from H5N1 highly pathogenic avian influenza A (HPAI) and Newcastle disease virus (NDV) can be achieved through vaccination, as part of a broader disease control strategy. We have previously generated a recombinant influenza virus expressing, (i) an H5 hemagglutinin protein, modified by the removal of the polybasic cleavage peptide and (ii) the ectodomain of the NDV hemagglutinin-neuraminidase (HN) protein in the place of the ectodomain of influenza neuraminidase (Park MS, et al. Proc Natl Acad Sci USA 2006;103(21):8203-8). Here we show this virus is attenuated in primary normal human bronchial epithelial (NHBE) cell culture, and demonstrate protection of C57BL/6 mice from lethal challenge with an H5 HA-containing influenza virus through immunisation with the recombinant virus. In addition, in-ovo vaccination of 18-day-old embryonated chicken eggs provided 90% and 80% protection against highly stringent lethal challenge by NDV and H5N1 virus, respectively. We propose that this virus has potential as a safe in-ovo live, attenuated, bivalent avian influenza and Newcastle disease virus vaccine.

Structure of a trimeric variant of the Epstein-Barr virus glycoprotein B

Energy Technology Data Exchange (ETDEWEB)

Backovic, Marija [Northwestern Univ., Evanston, IL (United States); Longnecker, Richard [Northwestern Univ., Chicago, IL (United States); Jardetzky, Theodore S [Northwestern Univ., Evanston, IL (United States)

2009-03-16

Epstein-Barr virus (EBV) is a herpesvirus that is associated with development of malignancies of lymphoid tissue. EBV infections are life-long and occur in >90% of the population. Herpesviruses enter host cells in a process that involves fusion of viral and cellular membranes. The fusion apparatus is comprised of envelope glycoprotein B (gB) and a heterodimeric complex made of glycoproteins H and L. Glycoprotein B is the most conserved envelope glycoprotein in human herpesviruses, and the structure of gB from Herpes simplex virus 1 (HSV-1) is available. Here, we report the crystal structure of the secreted EBV gB ectodomain, which forms 16-nm long spike-like trimers, structurally homologous to the postfusion trimers of the fusion protein G of vesicular stomatitis virus (VSV). Comparative structural analyses of EBV gB and VSV G, which has been solved in its pre and postfusion states, shed light on gB residues that may be involved in conformational changes and membrane fusion. Also, the EBV gB structure reveals that, despite the high sequence conservation of gB in herpesviruses, the relative orientations of individual domains, the surface charge distributions, and the structural details of EBV gB differ from the HSV-1 protein, indicating regions and residues that may have important roles in virus-specific entry.

E3L and F1L Gene Functions Modulate the Protective Capacity of Modified Vaccinia Virus Ankara Immunization in Murine Model of Human Smallpox

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Asisa Volz

2018-01-01

Full Text Available The highly attenuated Modified Vaccinia virus Ankara (MVA lacks most of the known vaccinia virus (VACV virulence and immune evasion genes. Today MVA can serve as a safety-tested next-generation smallpox vaccine. Yet, we still need to learn about regulatory gene functions preserved in the MVA genome, such as the apoptosis inhibitor genes F1L and E3L. Here, we tested MVA vaccine preparations on the basis of the deletion mutant viruses MVA-ΔF1L and MVA-ΔE3L for efficacy against ectromelia virus (ECTV challenge infections in mice. In non-permissive human tissue culture the MVA deletion mutant viruses produced reduced levels of the VACV envelope antigen B5. Upon mousepox challenge at three weeks after vaccination, MVA-ΔF1L and MVA-ΔE3L exhibited reduced protective capacity in comparison to wildtype MVA. Surprisingly, however, all vaccines proved equally protective against a lethal ECTV infection at two days after vaccination. Accordingly, the deletion mutant MVA vaccines induced high levels of virus-specific CD8+ T cells previously shown to be essential for rapidly protective MVA vaccination. These results suggest that inactivation of the anti-apoptotic genes F1L or E3L modulates the protective capacity of MVA vaccination most likely through the induction of distinct orthopoxvirus specific immunity in the absence of these viral regulatory proteins.

Gaseous NO2 effects on stomatal behavior, photosynthesis and respiration of hybrid poplar leaves

Science.gov (United States)

In this study, we used poplar as a model plant and investigated the effects of gaseous nitrogen dioxide (NO2, 4 microliter per liter) on stomatal conductance, photosynthesis, dark- and photorespiration of Populus alba x Populus berolinensis hybrid leaves using the photosynthesis system and scanning...

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

Crystallization and preliminary X-ray analysis of Chandipura virus glycoprotein G

International Nuclear Information System (INIS)

Baquero, Eduard; Buonocore, Linda; Rose, John K.; Bressanelli, Stéphane; Gaudin, Yves; Albertini, Aurélie A.

2012-01-01

Chandipura virus glycoprotein ectodomain (Gth) was purified and crystallized at pH 7.5. X-ray diffraction data set was collected to a resolution of 3.1 Å. Fusion in members of the Rhabdoviridae virus family is mediated by the G glycoprotein. At low pH, the G glycoprotein catalyzes fusion between viral and endosomal membranes by undergoing a major conformational change from a pre-fusion trimer to a post-fusion trimer. The structure of the G glycoprotein from vesicular stomatitis virus (VSV G), the prototype of Vesiculovirus, has recently been solved in its trimeric pre-fusion and post-fusion conformations; however, little is known about the structural details of the transition. In this work, a soluble form of the ectodomain of Chandipura virus G glycoprotein (CHAV G th ) was purified using limited proteolysis of purified virus; this soluble ectodomain was also crystallized. This protein shares 41% amino-acid identity with VSV G and thus its structure could provide further clues about the structural transition of rhabdoviral glycoproteins induced by low pH. Crystals of CHAV G th obtained at pH 7.5 diffracted X-rays to 3.1 Å resolution. These crystals belonged to the orthorhombic space group P2 1 2 1 2, with unit-cell parameters a = 150.3, b = 228.2, c = 78.8 Å. Preliminary analysis of the data based on the space group and the self-rotation function indicated that there was no trimeric association of the protomers. This unusual oligomeric status could result from the presence of fusion intermediates in the crystal

NIR-assisted orchid virus therapy using urchin bimetallic nanomaterials in phalaenopsis

International Nuclear Information System (INIS)

Chen, Shin-Yu; Do, Yi-Yin; Huang, Pung-Ling; Cheng, Liang-Chien; Chen, Chieh-Wei; Lee, Po-Han; Liu, Ru-Shi; Yu, Fengjiao; Zhou, Wuzong

2013-01-01

The use of nanoparticles has drawn special attention, particularly in the treatment of plant diseases. Cymbidium mosaic virus (CymMV) and Odontoglossum ring spot virus (ORSV) are the most prevalent and serious diseases that affect the development of the orchid industry. In this study we treated nanoparticles as a strategy for enhancing the resistance of orchids against CymMV and ORSV. After chitosan-modified gold nanoparticles (Au NPs) were injected into Phalaenopsis leaves, the injected leaves were exposed to 980 nm laser for light–heat conversion. To evaluate virus elimination in the treated Phalaenopsis leaves, the transcripts of coat protein genes and the production of viral proteins were assessed by reverse transcription-Polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. The expression of coat protein genes for both CymMV and ORSV was significantly lower in the chitosan-modified Au NP-treated Phalaenopsis leaves than in the control. Similarly, the amount of coat proteins for both viruses in the Phalaenopsis leaves was lower than that in the control (without nanoparticle injection). We propose that the temperature increase in the chitosan-modified Au NP-treated Phalaenopsis tissues after laser exposure reduces the viral population, consequently conferring resistance against CymMV and ORSV. Our findings suggest that the application of chitosan-modified Au NPs is a promising new strategy for orchid virus therapy. (paper)

Overexpression of StNF-YB3.1 reduces photosynthetic capacity and tuber production, and promotes ABA-mediated stomatal closure in potato (Solanum tuberosum L.).

Science.gov (United States)

Xuanyuan, Guochao; Lu, Congming; Zhang, Ruofang; Jiang, Jiming

2017-08-01

Nuclear factor Y (NF-Y) is one of the most ubiquitous transcription factors (TFs), comprising NF-YA, NF-YB and NF-YC subunits, and has been identified and reported in various aspects of development for plants and animals. In this work, StNF-YB3.1, a putative potato NF-YB subunit encoding gene, was isolated from Solanum tuberosum by rapid amplification of cDNA ends (RACE). Overexpression of StNF-YB3.1 in potato (cv. Atlantic) resulted in accelerated onset of flowering, and significant increase in leaf chlorophyll content in field trials. However, transgenic potato plants overexpressing StNF-YB3.1 (OEYB3.1) showed significant decreases in photosynthetic rate and stomatal conductance both at tuber initiation and bulking stages. OEYB3.1 lines were associated with significantly fewer tuber numbers and yield reduction. Guard cell size and stomatal density were not changed in OEYB3.1 plants, whereas ABA-mediated stomatal closure was accelerated compared to that of wild type plants because of the up-regulation of genes for ABA signaling, such as StCPK10-like, StSnRK2.6/OST1-like, StSnRK2.7-like and StSLAC1-like. We speculate that the acceleration of stomatal closure was a possible reason for the significantly decreased stomatal conductance and photosynthetic rate. Copyright © 2017 Elsevier B.V. All rights reserved.

Stomatal- and growth responses in willow to deficits in water- and nitrogen supply. Final report

Energy Technology Data Exchange (ETDEWEB)

Stadenberg, I. [Swedish Univ. of Agricultural Sciences, Uppsala (Sweden). Dep. for Production Ecology

2002-02-01

The two plants, grown with an [N] of 100 mg per litre and subjected to a decrease in N-supply decreased their leaf relative growth rate from 22% per day to 13% per day within 2 days. Stomatal conductance did not change significantly after the decrease in N-supply. Xylem samples did not show any significant changes in its composition of mineral nutrient elements after decreased N-supply. The three plants, grown with an [N] of 50 mg per litre and subjected to a decrease in N-supply, significantly decreased leaf relative growth rate from 18.5 % to 9 % per day within 2 days. Stomatal conductance did not change significantly after the decrease in N-supply. Xylem sap samples showed a significant decrease in [K] (74 mg/l to 42 mg/l) and [S] (11 mg/l to 3.2 mg/l) within 2 days after decreased N-supply. The four plants subjected to root drying decreased their leaf relative growth rate slightly but not significantly during the drying period. Xylem samples showed a significant decrease in S-concentration (11 mg/l to 1.3 mg/l) and [NO{sub 3}] (8.0 mg/l to 1.0 mg/l), while [Fe] increased significantly (0.065 mg/l to 0.14 mg/l). Stomatal conductance is known to decrease when plants are subjected to drying of part of the root system. This was shown for Salix dasyclados in a recent publication.

Serum cytokine profile and clinicopathological findings in oral lichen planus, oral lichenoid lesions and stomatitis

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Johansen, Jeanne Duus; Reibel, Jesper; Zachariae, Claus; Pedersen, Anne Marie Lynge

2017-01-01

Abstract 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‐nine patients and 29 healthy age‐ and gender‐matched subjects were included. Demographic and clinical data immunohistochemical findings in mucosal specimens, results of contact allergy testing, and serum levels of tumor necrosis factor‐α, interferon‐γ, interleukin (IL)‐6, IL‐10, IL‐12p40, and IL‐12p70 were analyzed and compared between groups. Nineteen patients had OLP, primarily with ulcerative lesions on the buccal mucosa, 19 patients had OLL, and 11 patients had generalized stomatitis. All patients had oral symptoms, mainly stinging and burning. Nineteen patients and 10 healthy subjects had contact allergies, primarily to fragrance ingredients. Patient groups did not differ with regard to oral symptoms, clinical pattern of the lesions, or contact allergy. Serum cytokine levels did not differ between the different patient groups and were not related to histopathological findings. The patients had higher levels of IL‐6 than the healthy subjects. Interferon‐γ, IL‐12p40, and IL‐12p70 were below detection limit. Our findings indicate that OLP, OLL, and generalized stomatitis cannot be discriminated by means of the selected serum cytokines, and that the presence of concomitant contact allergy does not influence the cytokine expression. PMID:29744205

Detecting the differences in responses of stomatal conductance to moisture stresses between deciduous shrubs and Artemisia subshrubs.

Science.gov (United States)

Gao, Qiong; Yu, Mei; Zhou, Chan

2013-01-01

Shrubs and subshrubs can tolerate wider ranges of moisture stresses in both soil and air than other plant life forms, and thus represent greater nonlinearity and uncertainty in ecosystem physiology. The objectives of this paper are to model shrub/subshrub stomatal conductance by synthesizing the field leaf gas exchanges data of 24 species in China, in order to detect the differences between deciduous shrubs and Artemisia subshrubs in their responses of stomatal conductance to changes in the moisture stresses. We revised a model of stomatal conductance by incorporating the tradeoff between xylem hydraulic efficiency and cavitation loss risk. We then fit the model at the three hierarchical levels: global (pooling all data as a single group), three functional groups (deciduous non-legume shrubs, deciduous legume shrubs, and subshrubs in Artemisia genus), and individual observations (species × sites). Bayesian inference with Markov Chain Monte Carlo method was applied to obtain the model parameters at the three levels. We found that the model at the level of functional groups is a significant improvement over that at the global level, indicating the significant differences in the stomatal behavior among the three functional groups. The differences in tolerance and sensitivities to changes in moisture stresses are the most evident between the shrubs and the subshrubs: The two shrub groups can tolerate much higher soil water stress than the subshrubs. The analysis at the observation level is also a significant improvement over that at the functional group level, indicating great variations within each group. Our analysis offered a clue for the equivocal issue of shrub encroachment into grasslands: While the invasion by the shrubs may be irreversible, the dominance of subshrubs, due to their lower resistance and tolerance to moisture stresses, may be put down by appropriate grassland management.

Use of sap flow measurements to validate stomatal functions for mature beech (Fagus sylvatica) in view of ozone uptake calculations

International Nuclear Information System (INIS)

Braun, Sabine; Schindler, Christian; Leuzinger, Sebastian

2010-01-01

For a quantitative estimate of the ozone effect on vegetation reliable models for ozone uptake through the stomata are needed. Because of the analogy of ozone uptake and transpiration it is possible to utilize measurements of water loss such as sap flow for quantification of ozone uptake. This technique was applied in three beech (Fagus sylvatica) stands in Switzerland. A canopy conductance was calculated from sap flow velocity and normalized to values between 0 and 1. It represents mainly stomatal conductance as the boundary layer resistance in forests is usually small. Based on this relative conductance, stomatal functions to describe the dependence on light, temperature, vapour pressure deficit and soil moisture were derived using multivariate nonlinear regression. These functions were validated by comparison with conductance values directly estimated from sap flow. The results corroborate the current flux parameterization for beech used in the DO 3 SE model. - A method was developed to derive stomatal functions and ozone uptake calculation from sap flow.

Au nanocrystals grown on a better-defined one-dimensional tobacco mosaic virus coated protein template genetically modified by a hexahistidine tag

International Nuclear Information System (INIS)

Liu Nan; Zhang Wei; Luo Zhaopeng; Zhai Niu; Zhang Hongfei; Li Zhonghao; Jiang Xingyi; Tang Gangling; Hu Qingyuan; Wang Chong; Tian Dandan

2012-01-01

In this paper, tobacco mosaic virus (TMV) coated protein (CP) was genetically modified by introducing a hexahistidine tag into it for a well-defined one-dimensional template, on which Au nanocrystals (NCs) were grown. The results showed that genetic modification could not only ameliorate the one-dimensional structure of the template, but also improve the growth density of Au NCs on the template. This indicated that genetic modification could be an effective method to modulate the structure of the TMVCP template-based nanocomposites allowing for a broader application of them. (paper)

Maximum leaf conductance driven by CO2 effects on stomatal size and density over geologic time.

Science.gov (United States)

Franks, Peter J; Beerling, David J

2009-06-23

Stomatal pores are microscopic structures on the epidermis of leaves formed by 2 specialized guard cells that control the exchange of water vapor and CO(2) between plants and the atmosphere. Stomatal size (S) and density (D) determine maximum leaf diffusive (stomatal) conductance of CO(2) (g(c(max))) to sites of assimilation. Although large variations in D observed in the fossil record have been correlated with atmospheric CO(2), the crucial significance of similarly large variations in S has been overlooked. Here, we use physical diffusion theory to explain why large changes in S necessarily accompanied the changes in D and atmospheric CO(2) over the last 400 million years. In particular, we show that high densities of small stomata are the only way to attain the highest g(cmax) values required to counter CO(2)"starvation" at low atmospheric CO(2) concentrations. This explains cycles of increasing D and decreasing S evident in the fossil history of stomata under the CO(2) impoverished atmospheres of the Permo-Carboniferous and Cenozoic glaciations. The pattern was reversed under rising atmospheric CO(2) regimes. Selection for small S was crucial for attaining high g(cmax) under falling atmospheric CO(2) and, therefore, may represent a mechanism linking CO(2) and the increasing gas-exchange capacity of land plants over geologic time.

Avian metapneumovirus RT-nested-PCR: a novel false positive reducing inactivated control virus with potential applications to other RNA viruses and real time methods.

Science.gov (United States)

Falchieri, Marco; Brown, Paul A; Catelli, Elena; Naylor, Clive J

2012-12-01

Using reverse genetics, an avian metapneumovirus (AMPV) was modified for use as a positive control for validating all stages of a popular established RT-nested PCR, used in the detection of the two major AMPV subtypes (A and B). Resultant amplicons were of increased size and clearly distinguishable from those arising from unmodified virus, thus allowing false positive bands, due to control virus contamination of test samples, to be identified readily. Absorption of the control virus onto filter paper and subsequent microwave irradiation removed all infectivity while its function as an efficient RT-nested-PCR template was unaffected. Identical amplicons were produced after storage for one year. The modified virus is likely to have application as an internal standard as well as in real time methods. Additions to AMPV of RNA from other RNA viruses, including hazardous examples such HIV and influenza, are likely to yield similar safe RT-PCR controls. Copyright © 2012 Elsevier B.V. All rights reserved.

Three-year rabies duration of immunity in dogs following vaccination with a core combination vaccine against canine distemper virus, canine adenovirus type-1, canine parvovirus, and rabies virus.

Science.gov (United States)

Lakshmanan, Nallakannu; Gore, Thomas C; Duncan, Karen L; Coyne, Michael J; Lum, Melissa A; Sterner, Frank J

2006-01-01

Thirty-two seronegative pups were vaccinated at 8 weeks of age with modified-live canine distemper virus (CDV), canine adenovirus type-2 (CAV-2), and canine parvovirus (CPV) vaccine and at 12 weeks with a modified-live CDV, CAV-2, CPV, and killed rabies virus vaccine. An additional 31 seronegative pups served as age-matched, nonvaccinated controls. All test dogs were strictly isolated for 3 years after receiving the second vaccination and then were challenged with virulent rabies virus. Clinical signs of rabies were prevented in 28 (88%) of the 32 vaccinated dogs. In contrast, 97% (30 of 31) of the control dogs died of rabies infection. These study results indicated that no immunogenic interference occurred between the modified-live vaccine components and the killed rabies virus component. Furthermore, these results indicated that the rabies component in the test vaccine provided protection against virulent rabies challenge in dogs 12 weeks of age or older for a minimum of 3 years following vaccination.

Brief oral cryotherapy for the prevention of high-dose melphalan-induced stomatitis in allogeneic hematopoietic stem cell transplant recipients.

Science.gov (United States)

Mori, Takehiko; Yamazaki, Rie; Aisa, Yoshinobu; Nakazato, Tomonori; Kudo, Masumi; Yashima, Tomoko; Kondo, Sakiko; Ikeda, Yasuo; Okamoto, Shinichiro

2006-04-01

We previously reported the efficacy of oral cryotherapy for the prevention of high-dose melphalan-induced stomatitis. The purpose of this study was to evaluate whether the further shortening of the duration of oral cryotherapy could minimize its side effects while sparing its efficacy. Seventeen consecutive recipients of allogeneic hematopoieic stem cell transplant conditioned with high-dose melphalan in combination with fludarabine alone or with fludarabine and additional radiation were enrolled in the study. The severity of stomatitis was graded according to the National Cancer Institute-Common Toxicity Criteria. Patients were kept on oral cryotherapy shortly before, during, and for additional 30 min after the completion of melphalan administration (60-min oral cryotherapy). Patients who were also enrolled in our previous study received the same type of oral cryotherapy but for additional 90 min after the completion of melphalan administration (120-min oral cryotherapy), and they served as controls. Only 2 (11.8%) of 17 patients receiving 60-min oral cryotherapy and 2 (11.1%) of 18 patients receiving 120-min oral cryotherapy developed grade 2 or 3 stomatitis, respectively. The difference between groups was not statistically significant (P = 0.677). The incidence of unpleasant symptoms such as chills and nausea during oral cryotherapy decreased significantly with 60-min oral cryotherapy, as compared with that associated with 120-min oral cryotherapy (P cryotherapy is as effective as 120-min oral cryotherapy at preventing high-dose melphalan-induced stomatitis, and shorter treatment might have contributed to relieve patient discomfort during oral cryotherapy.

Quantitative Estimation of Yeast on Maxillary Denture in Patients with Denture Stomatitis and the Effect of Chlorhexidine Gluconate in Reduction of Yeast

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Jaykumar R Gade

2011-01-01

Full Text Available Denture stomatitis is a condition associated with wearing of a denture. The predisposing factor leading to denture stomatitis could be poor oral hygiene, ill-fitting denture and relief areas. Around 30 patients with denture stomatitis were advised to rinse with chlorhexidine gluconate mouthwash for 14 days and were directed to immerse the upper denture in the chlorhexidine solution for 8 hours. The samples were collected by scraping maxillary denture in saline at three intervals, prior to, at the end of 24 hours and after 14 days of treatment, then were inoculated and quantitative estimation of the yeast growth on Sabouraud′s dextrose agar plate was done. It was observed that after a period of 14 days, there was a reduction in the growth of yeast and also improvement in the clinical picture of the oral mucosa

Vaccination with recombinant RNA replicon particles protects chickens from H5N1 highly pathogenic avian influenza virus.

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Stefan J Halbherr

Full Text Available Highly pathogenic avian influenza viruses (HPAIV of subtype H5N1 not only cause a devastating disease in domestic chickens and turkeys but also pose a continuous threat to public health. In some countries, H5N1 viruses continue to circulate and evolve into new clades and subclades. The rapid evolution of these viruses represents a problem for virus diagnosis and control. In this work, recombinant vesicular stomatitis virus (VSV vectors expressing HA of subtype H5 were generated. To comply with biosafety issues the G gene was deleted from the VSV genome. The resulting vaccine vector VSV*ΔG(HA was propagated on helper cells providing the VSV G protein in trans. Vaccination of chickens with a single intramuscular dose of 2×10⁸ infectious replicon particles without adjuvant conferred complete protection from lethal H5N1 infection. Subsequent application of the same vaccine strongly boosted the humoral immune response and completely prevented shedding of challenge virus and transmission to sentinel birds. The vaccine allowed serological differentiation of infected from vaccinated animals (DIVA by employing a commercially available ELISA. Immunized chickens produced antibodies with neutralizing activity against multiple H5 viruses representing clades 1, 2.2, 2.5, and low-pathogenic avian influenza viruses (classical clade. Studies using chimeric H1/H5 hemagglutinins showed that the neutralizing activity was predominantly directed against the globular head domain. In summary, these results suggest that VSV replicon particles are safe and potent DIVA vaccines that may help to control avian influenza viruses in domestic poultry.

Environmental controls on saltcedar (Tamarix spp.) transpiration and stomatal conductance and implications for determining evapotranspiration by remote sensing

Science.gov (United States)

Nagler, P. L.; Glenn, E. P.; morino, K.

2012-12-01

Saltcedar is an introduced, salt-tolerant shrub that now dominates many flow-regulated western U.S. rivers. Saltcedar control programs have been implemented to salvage water and to allow the return of native vegetation to infested rivers. However, there is much debate about how much water saltcedar actually uses and the range of ecohydrological niches it occupies. Ground methods for measuring riparian zone ET have improved and there is considerable interest in developing remote sensing methods for saltcedar to conduct wide-area monitoring of water use. Both thermal band and vegetation index methods have been used to estimate riparian ET. However, several problems present themselves in applying existing remote sensing methods to riparian corridors. First, many riparian corridors are narrow and are surrounded by arid uplands, hence they cannot be treated as energetically closed systems, an assumption of thermal band methods that calculate ET as a residual in the surface energy balance. Second, contrary to the assumption that riparian phreatophytes typically grow under unstressed conditions since they are rooted into groundwater, we find that saltcedar stands are under substantial degrees of apparent moisture stress, exhibiting midday depression of transpiration and stomatal conductance, and decreases in stomatal conductance over the growing season as depth to groundwater increases. Furthermore, the degree of stress is site-specific, depending on local soil texture, salinity of the groundwater and distance from the river. This violates a key assumption of vegetation index methods for estimating ET. The implications of these findings for arid-zone riparian ecohydrology and for remote sensing methods that assume either a constant daily evaporative fraction or rate of stomatal conductance will be discussed using saltcedar stands measured in the Cibola NWR on the lower Colorado River as a case study. Daily rates of saltcedar transpiration ranged from 1.6-3.0 mm/m2 leaf

RIN4 functions with plasma membrane H+-ATPases to regulate stomatal apertures during pathogen attack

DEFF Research Database (Denmark)

Liu, Jun; Elmore, James M.; Fuglsang, Anja Thoe

2009-01-01

Abstract Pathogen perception by the plant innate immune system is of central importance to plant survival and productivity. The Arabidopsis protein RIN4 is a negative regulator of plant immunity. In order to identify additional proteins involved in RIN4- mediated immune signal transduction, we...... purified components of the RIN4 protein complex. We identified six novel proteins that had not previously been implicated in RIN4 signaling, including the plasma membrane (PM) H+-ATPases AHA1 and/or AHA2. RIN4 interacts with AHA1 and AHA2 both in vitro and in vivo. RIN4 overexpression and knockout lines...... exhibit differential PM H+-ATPase activity. PM H+-ATPase activation induces stomatal opening, enabling bacteria to gain entry into the plant leaf; inactivation induces stomatal closure thus restricting bacterial invasion. The rin4 knockout line exhibited reduced PM H+-ATPase activity and, importantly, its...

Water treatment with exceptional virus inactivation using activated carbon modified with silver (Ag) and copper oxide (CuO) nanoparticles.

Science.gov (United States)

Shimabuku, Quelen Letícia; Arakawa, Flávia Sayuri; Fernandes Silva, Marcela; Ferri Coldebella, Priscila; Ueda-Nakamura, Tânia; Fagundes-Klen, Márcia Regina; Bergamasco, Rosangela

2017-08-01

Continuous flow experiments (450 mL min -1 ) were performed in household filter in order to investigate the removal and/or inactivation of T4 bacteriophage, using granular activated carbon (GAC) modified with silver and/or copper oxide nanoparticles at different concentrations. GAC and modified GAC were characterized by X-ray diffractometry, specific surface area, pore size and volume, pore average diameter, scanning electron microscopy, transmission electron microscopy, zeta potential and atomic absorption spectroscopy. The antiviral activity of the produced porous media was evaluated by passing suspensions of T4 bacteriophage (∼10 5  UFP/mL) through filters. The filtered water was analyzed for the presence of the bacteriophage and the release of silver and copper oxide. The porous media containing silver and copper oxide nanoparticles showed high inactivation capacity, even reaching reductions higher than 3 log. GAC6 (GAC/Ag0.5%Cu1.0%) was effective in the bacteriophage inactivation, reaching 5.53 log reduction. The levels of silver and copper released in filtered water were below the recommended limits (100 ppb for silver and 1000 ppb for copper) in drinking water. From this study, it is possible to conclude that activated carbon modified with silver and copper oxide nanoparticles can be used as a filter for virus removal in the treatment of drinking water.

Recurrent aphthous stomatitis (RAS and exfoliative cheilitis in elderly psoriasis sufferer

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Siti Hardiyanti Nurhasanah

2016-04-01

Full Text Available Recurrent Aphthous Stomatitis (RAS is a disorder in the oral cavity, with a characterized symptom as ulceration, recurrent and very painfull. The etiology is idiopathic, with multifactorial predisposition. Exfoliative cheilitis is a persistent lesion on the lip, with a characterized cracking and desquamative, with crustae and inflammation. An elderly male (72 yrs suffered with ulcer on his oral cavity, cracking lips and pain on both of his cheeks, skin, since 5 years ago. The pain is recurrent. On the clinical examination, there were some desquamation, both on the skin and vermilion border, whether on the inner lips (labial fold mucosa, there were ulcers with diameter about 1 cm. The laboratory test was within normal limits, except the LED was 40 mm/hour (n:<15. The diagnosis was Recurrent Aphthous Stomatitis (RAS for the ulcer and Exfoliative cheilitis for the cracking lips. The treatment he received was a gargle liquid, topical corticosteroid and supplement. The skin’s disorder was revered to the skin and genital disease department, for further management. As a dental general practioner, had to be very careful and familiar for every changes that may be occur both in the outer or inner oral cavity. Other disorder that need refferal, had to be done with team work, to the colleague from the right connection.

SAT2 Foot-and-Mouth Disease Virus Structurally Modified for Increased Thermostability.

Science.gov (United States)

Scott, Katherine A; Kotecha, Abhay; Seago, Julian; Ren, Jingshan; Fry, Elizabeth E; Stuart, David I; Charleston, Bryan; Maree, Francois F

2017-05-15

Foot-and-mouth disease virus (FMDV), particularly strains of the O and SAT serotypes, is notoriously unstable. Consequently, vaccines derived from heat-labile SAT viruses have been linked to the induction of immunity with a poor duration and hence require more frequent vaccinations to ensure protection. In silico calculations predicted residue substitutions that would increase interactions at the interpentamer interface, supporting increased stability. We assessed the stability of the 18 recombinant mutant viruses in regard to their growth kinetics, antigenicity, plaque morphology, genetic stability, and temperature, ionic, and pH stability by using Thermofluor and inactivation assays in order to evaluate potential SAT2 vaccine candidates with improved stability. The most stable mutant for temperature and pH stability was the S2093Y single mutant, while other promising mutants were the E3198A, L2094V, and S2093H single mutants and the F2062Y-H2087M-H3143V triple mutant. Although the S2093Y mutant had the greatest stability, it exhibited smaller plaques, a reduced growth rate, a change in monoclonal antibody footprint, and poor genetic stability properties compared to those of the wild-type virus. However, these factors affecting production can be overcome. The addition of 1 M NaCl was found to further increase the stability of the SAT2 panel of viruses. The S2093Y and S2093H mutants were selected for future use in stabilizing SAT2 vaccines. IMPORTANCE Foot-and-mouth disease virus (FMDV) causes a highly contagious acute vesicular disease in cloven-hoofed livestock and wildlife. The control of the disease by vaccination is essential, especially at livestock-wildlife interfaces. The instability of some serotypes, such as SAT2, affects the quality of vaccines and therefore the duration of immunity. We have shown that we can improve the stability of SAT2 viruses by mutating residues at the capsid interface through predictive modeling. This is an important finding for

Psoralen-mediated virus photoinactivation in platelet concentrates: enhanced specificity of virus kill in the absence of shorter UVA wavelengths

International Nuclear Information System (INIS)

Margolis-Nunno, Henrietta; Robinson, Richard; Horowitz, Bernard; Ben-Hur, Ehud; Geacintov, N.E.

1995-01-01

Treatments with psoralens and long-wavelength ultraviolet radiation (UVA, 320-400 nm; PUVA) have shown efficacy for virus sterilization of platelet concentrates (PC). We have employed the psoralen derivative 4'-aminomethyl-4,5',8-trimethylpsoralen (AMT), and have found that platelet integrity is best preserved when rutin, a flavonoid that quenches multiple reactive oxygen species, is present during AMT/UVA treatment of PC. In this report, we examine the effects of different UVA spectra under our standard PC treatment conditions (i.e. 50 μg/mL AMT, 0.35 mM rutin and 38 J/cm 2 UVA). Added vesicular stomatitis virus (VSV; ≥ 5.5 log 10 ) was completely inactivated with the simultaneous maintenance of the platelet aggregation response (> 90% of control) when a UVA light source with transmission mainly between 360 and 370 nm (narrow UVA1) was used. In contrast, with a broad-band UVA (320-400 nm; broad UVA) light source, the aggregation response was greatly compromised (< 50% of control) with only a minor increase in the rate of VSV kill. With this lamp, platelet function could be improved to about 75% of the control by adding a long-pass filter, which reduced the transmission of shorter (≤ 345 nm) UVA wavelengths (340-400 nm; UVA1). At equivalent levels of virus kill, aggregation function was always best preserved when narrow UVA1 was used for PUVA treatment. Even in the absence of AMT, and with or without rutin present, narrow UVA1 irradiation was better tolerated by platelets than was broad UVA. (author)

Mucosal immunization with PLGA-microencapsulated DNA primes a SIV-specific CTL response revealed by boosting with cognate recombinant modified vaccinia virus Ankara

International Nuclear Information System (INIS)

Sharpe, Sally; Hanke, Tomas; Tinsley-Bown, Anne; Dennis, Mike; Dowall, Stuart; McMichael, Andrew; Cranage, Martin

2003-01-01

Systemically administered DNA encoding a recombinant human immunodeficiency virus (HIV) derived immunogen effectively primes a cytotoxic T lymphocyte (CTL) response in macaques. In this further pilot study we have evaluated mucosal delivery of DNA as an alternative priming strategy. Plasmid DNA, pTH.HW, encoding a multi-CTL epitope gene, was incorporated into poly(D,L-lactic-co-glycolic acid) microparticles of less than 10 μm in diameter. Five intrarectal immunizations failed to stimulate a circulating vaccine-specific CTL response in 2 Mamu-A*01 + rhesus macaques. However, 1 week after intradermal immunization with a cognate modified vaccinia virus Ankara vaccine MVA.HW, CTL responses were detected in both animals that persisted until analysis postmortem, 12 weeks after the final boost. In contrast, a weaker and less durable response was seen in an animal vaccinated with the MVA construct alone. Analysis of lymphoid tissues revealed a disseminated CTL response in peripheral and regional lymph nodes but not the spleen of both mucosally primed animals

Inhibition of influenza virus infection and hemagglutinin cleavage by the protease inhibitor HAI-2

Energy Technology Data Exchange (ETDEWEB)

Hamilton, Brian S.; Chung, Changik; Cyphers, Soreen Y.; Rinaldi, Vera D.; Marcano, Valerie C.; Whittaker, Gary R., E-mail: grw7@cornell.edu

2014-07-25

Highlights: • Biochemical and cell biological analysis of HAI-2 as an inhibitor of influenza HA cleavage activation. • Biochemical and cell biological analysis of HAI-2 as an inhibitor of influenza virus infection. • Comparative analysis of HAI-2 for vesicular stomatitis virus and human parainfluenza virus type-1. • Analysis of the activity of HAI-2 in a mouse model of influenza. - Abstract: Influenza virus remains a significant concern to public health, with the continued potential for a high fatality pandemic. Vaccination and antiviral therapeutics are effective measures to circumvent influenza virus infection, however, multiple strains have emerged that are resistant to the antiviral therapeutics currently on the market. With this considered, investigation of alternative antiviral therapeutics is being conducted. One such approach is to inhibit cleavage activation of the influenza virus hemagglutinin (HA), which is an essential step in the viral replication cycle that permits viral-endosome fusion. Therefore, targeting trypsin-like, host proteases responsible for HA cleavage in vivo may prove to be an effective therapeutic. Hepatocyte growth factor activator inhibitor 2 (HAI-2) is naturally expressed in the respiratory tract and is a potent inhibitor of trypsin-like serine proteases, some of which have been determined to cleave HA. In this study, we demonstrate that HAI-2 is an effective inhibitor of cleavage of HA from the human-adapted H1 and H3 subtypes. HAI-2 inhibited influenza virus H1N1 infection in cell culture, and HAI-2 administration showed protection in a mouse model of influenza. HAI-2 has the potential to be an effective, alternative antiviral therapeutic for influenza.

Inhibition of influenza virus infection and hemagglutinin cleavage by the protease inhibitor HAI-2

International Nuclear Information System (INIS)

Hamilton, Brian S.; Chung, Changik; Cyphers, Soreen Y.; Rinaldi, Vera D.; Marcano, Valerie C.; Whittaker, Gary R.

2014-01-01

Highlights: • Biochemical and cell biological analysis of HAI-2 as an inhibitor of influenza HA cleavage activation. • Biochemical and cell biological analysis of HAI-2 as an inhibitor of influenza virus infection. • Comparative analysis of HAI-2 for vesicular stomatitis virus and human parainfluenza virus type-1. • Analysis of the activity of HAI-2 in a mouse model of influenza. - Abstract: Influenza virus remains a significant concern to public health, with the continued potential for a high fatality pandemic. Vaccination and antiviral therapeutics are effective measures to circumvent influenza virus infection, however, multiple strains have emerged that are resistant to the antiviral therapeutics currently on the market. With this considered, investigation of alternative antiviral therapeutics is being conducted. One such approach is to inhibit cleavage activation of the influenza virus hemagglutinin (HA), which is an essential step in the viral replication cycle that permits viral-endosome fusion. Therefore, targeting trypsin-like, host proteases responsible for HA cleavage in vivo may prove to be an effective therapeutic. Hepatocyte growth factor activator inhibitor 2 (HAI-2) is naturally expressed in the respiratory tract and is a potent inhibitor of trypsin-like serine proteases, some of which have been determined to cleave HA. In this study, we demonstrate that HAI-2 is an effective inhibitor of cleavage of HA from the human-adapted H1 and H3 subtypes. HAI-2 inhibited influenza virus H1N1 infection in cell culture, and HAI-2 administration showed protection in a mouse model of influenza. HAI-2 has the potential to be an effective, alternative antiviral therapeutic for influenza

Evolution of the bHLH genes involved in stomatal development: implications for the expansion of developmental complexity of stomata in land plants.

Directory of Open Access Journals (Sweden)

Jin-Hua Ran

Full Text Available Stomata play significant roles in plant evolution. A trio of closely related basic Helix-Loop-Helix (bHLH subgroup Ia genes, SPCH, MUTE and FAMA, mediate sequential steps of stomatal development, and their functions may be conserved in land plants. However, the evolutionary history of the putative SPCH/MUTE/FAMA genes is still greatly controversial, especially the phylogenetic positions of the bHLH Ia members from basal land plants. To better understand the evolutionary pattern and functional diversity of the bHLH genes involved in stomatal development, we made a comprehensive evolutionary analysis of the homologous genes from 54 species representing the major lineages of green plants. The phylogenetic analysis indicated: (1 All bHLH Ia genes from the two basal land plants Physcomitrella and Selaginella were closely related to the FAMA genes of seed plants; and (2 the gymnosperm 'SPCH' genes were sister to a clade comprising the angiosperm SPCH and MUTE genes, while the FAMA genes of gymnosperms and angiosperms had a sister relationship. The revealed phylogenetic relationships are also supported by the distribution of gene structures and previous functional studies. Therefore, we deduce that the function of FAMA might be ancestral in the bHLH Ia subgroup. In addition, the gymnosperm "SPCH" genes may represent an ancestral state and have a dual function of SPCH and MUTE, two genes that could have originated from a duplication event in the common ancestor of angiosperms. Moreover, in angiosperms, SPCHs have experienced more duplications and harbor more copies than MUTEs and FAMAs, which, together with variation of the stomatal development in the entry division, implies that SPCH might have contributed greatly to the diversity of stomatal development. Based on the above, we proposed a model for the correlation between the evolution of stomatal development and the genes involved in this developmental process in land plants.

GID1 modulates stomatal response and submergence tolerance involving abscisic acid and gibberellic acid signaling in rice.

Science.gov (United States)

Du, Hao; Chang, Yu; Huang, Fei; Xiong, Lizhong

2015-11-01

Plant responses to abiotic stresses are coordinated by arrays of growth and developmental programs. Gibberellic acid (GA) and abscisic acid (ABA) play critical roles in the developmental programs and environmental responses, respectively, through complex signaling and metabolism networks. However, crosstalk between the two phytohormones in stress responses remains largely unknown. In this study, we report that GIBBERELLIN-INSENSITIVE DWARF 1 (GID1), a soluble receptor for GA, regulates stomatal development and patterning in rice (Oryza sativa L.). The gid1 mutant showed impaired biosynthesis of endogenous ABA under drought stress conditions, but it exhibited enhanced sensitivity to exogenous ABA. Scanning electron microscope and infrared thermal image analysis indicated an increase in the stomatal conductance in the gid1 mutant under drought conditions. Interestingly, the gid1 mutant had increased levels of chlorophyll and carbohydrates under submergence conditions, and showed enhanced reactive oxygen species (ROS)-scavenging ability and submergence tolerance compared with the wild-type. Further analyses suggested that the function of GID1 in submergence responses is partially dependent on ABA, and GA signaling by GID1 is involved in submergence tolerance by modulating carbohydrate consumption. Taken together, these findings suggest GID1 plays distinct roles in stomatal response and submergence tolerance through both the ABA and GA signaling pathways in rice. © 2014 Institute of Botany, Chinese Academy of Sciences.

Early transduction produces highly functional chimeric antigen receptor-modified virus-specific T-cells with central memory markers: a Production Assistant for Cell Therapy (PACT) translational application.

Science.gov (United States)

Sun, Jiali; Huye, Leslie E; Lapteva, Natalia; Mamonkin, Maksim; Hiregange, Manasa; Ballard, Brandon; Dakhova, Olga; Raghavan, Darshana; Durett, April G; Perna, Serena K; Omer, Bilal; Rollins, Lisa A; Leen, Ann M; Vera, Juan F; Dotti, Gianpietro; Gee, Adrian P; Brenner, Malcolm K; Myers, Douglas G; Rooney, Cliona M

2015-01-01

Virus-specific T-cells (VSTs) proliferate exponentially after adoptive transfer into hematopoietic stem cell transplant (HSCT) recipients, eliminate virus infections, then persist and provide long-term protection from viral disease. If VSTs behaved similarly when modified with tumor-specific chimeric antigen receptors (CARs), they should have potent anti-tumor activity. This theory was evaluated by Cruz et al. in a previous clinical trial with CD19.CAR-modified VSTs, but there was little apparent expansion of these cells in patients. In that study, VSTs were gene-modified on day 19 of culture and we hypothesized that by this time, sufficient T-cell differentiation may have occurred to limit the subsequent proliferative capacity of the transduced T-cells. To facilitate the clinical testing of this hypothesis in a project supported by the NHLBI-PACT mechanism, we developed and optimized a good manufacturing practices (GMP) compliant method for the early transduction of VSTs directed to Epstein-Barr virus (EBV), Adenovirus (AdV) and cytomegalovirus (CMV) using a CAR directed to the tumor-associated antigen disialoganglioside (GD2). Ad-CMVpp65-transduced EBV-LCLs effectively stimulated VSTs directed to all three viruses (triVSTs). Transduction efficiency on day three was increased in the presence of cytokines and high-speed centrifugation of retroviral supernatant onto retronectin-coated plates, so that under optimal conditions up to 88% of tetramer-positive VSTs expressed the GD2.CAR. The average transduction efficiency of early-and late transduced VSTs was 55 ± 4% and 22 ± 5% respectively, and early-transduced VSTs maintained higher frequencies of T cells with central memory or intermediate memory phenotypes. Early-transduced VSTs also had higher proliferative capacity and produced higher levels of TH1 cytokines IL-2, TNF-α, IFN-γ, MIP-1α, MIP-1β and other cytokines in vitro. We developed a rapid and GMP compliant method for the early transduction of

Disposal of Hospital Wastes Containing Pathogenic Organisms

Science.gov (United States)

1979-09-01

virus African swine fever virus Besnoitia besnoiti Borna disease virus Bovine infectious petechial fever virus Camel pox virus Ephemeral fever virus...Sindbis virus Tensaw virus Turlock virus Vaccinia virus Varicella virus Vole rickettsia Yellow fever virus, 17D vaccinL strain 163 Class 3 AlastruLn...Rickettsia - all species except Vole rickettsia when used for transmission or animal inoculation experiments Vesicular stomatitis virus Yellow fever virus

A novel bHLH transcription factor PebHLH35 from Populus euphratica confers drought tolerance through regulating stomatal development, photosynthesis and growth in Arabidopsis

Energy Technology Data Exchange (ETDEWEB)

Dong, Yan [College of Biological Sciences and Technology, National Engineering Laboratory for Tree Breeding, Beijing Forestry University, Beijing 100083 (China); Liaoning Forestry Vocational-Technical College, Shenyang 110101 (China); Wang, Congpeng; Han, Xiao; Tang, Sha; Liu, Sha [College of Biological Sciences and Technology, National Engineering Laboratory for Tree Breeding, Beijing Forestry University, Beijing 100083 (China); Xia, Xinli, E-mail: xiaxl@bjfu.edu.cn [College of Biological Sciences and Technology, National Engineering Laboratory for Tree Breeding, Beijing Forestry University, Beijing 100083 (China); Yin, Weilun, E-mail: yinwl@bjfu.edu.cn [College of Biological Sciences and Technology, National Engineering Laboratory for Tree Breeding, Beijing Forestry University, Beijing 100083 (China)

2014-07-18

Highlights: • PebHLH35 is firstly cloned from Populus euphratica and characterized its functions. • PebHLH35 is important for earlier seedling establishment and vegetative growth. • PebHLH35 enhances tolerance to drought by regulating growth. • PebHLH35 enhances tolerance to drought by regulating stomatal development. • PebHLH35 enhances tolerance to drought by regulating photosynthesis and transpiration. - Abstract: Plant basic helix-loop-helix (bHLH) transcription factors (TFs) are involved in a variety of physiological processes including the regulation of plant responses to various abiotic stresses. However, few drought-responsive bHLH family members in Populus have been reported. In this study, a novel bHLH gene (PebHLH35) was cloned from Populus euphratica. Expression analysis in P. euphratica revealed that PebHLH35 was induced by drought and abscisic acid. Subcellular localization studies using a PebHLH35-GFP fusion showed that the protein was localized to the nucleus. Ectopic overexpression of PebHLH35 in Arabidopsis resulted in a longer primary root, more leaves, and a greater leaf area under well-watered conditions compared with vector control plants. Notably, PebHLH35 overexpression lines showed enhanced tolerance to water-deficit stress. This finding was supported by anatomical and physiological analyses, which revealed a reduced stomatal density, stomatal aperture, transpiration rate, and water loss, and a higher chlorophyll content and photosynthetic rate. Our results suggest that PebHLH35 functions as a positive regulator of drought stress responses by regulating stomatal density, stomatal aperture, photosynthesis and growth.

A novel bHLH transcription factor PebHLH35 from Populus euphratica confers drought tolerance through regulating stomatal development, photosynthesis and growth in Arabidopsis

International Nuclear Information System (INIS)

Dong, Yan; Wang, Congpeng; Han, Xiao; Tang, Sha; Liu, Sha; Xia, Xinli; Yin, Weilun

2014-01-01

Highlights: • PebHLH35 is firstly cloned from Populus euphratica and characterized its functions. • PebHLH35 is important for earlier seedling establishment and vegetative growth. • PebHLH35 enhances tolerance to drought by regulating growth. • PebHLH35 enhances tolerance to drought by regulating stomatal development. • PebHLH35 enhances tolerance to drought by regulating photosynthesis and transpiration. - Abstract: Plant basic helix-loop-helix (bHLH) transcription factors (TFs) are involved in a variety of physiological processes including the regulation of plant responses to various abiotic stresses. However, few drought-responsive bHLH family members in Populus have been reported. In this study, a novel bHLH gene (PebHLH35) was cloned from Populus euphratica. Expression analysis in P. euphratica revealed that PebHLH35 was induced by drought and abscisic acid. Subcellular localization studies using a PebHLH35-GFP fusion showed that the protein was localized to the nucleus. Ectopic overexpression of PebHLH35 in Arabidopsis resulted in a longer primary root, more leaves, and a greater leaf area under well-watered conditions compared with vector control plants. Notably, PebHLH35 overexpression lines showed enhanced tolerance to water-deficit stress. This finding was supported by anatomical and physiological analyses, which revealed a reduced stomatal density, stomatal aperture, transpiration rate, and water loss, and a higher chlorophyll content and photosynthetic rate. Our results suggest that PebHLH35 functions as a positive regulator of drought stress responses by regulating stomatal density, stomatal aperture, photosynthesis and growth

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...... potentially enhance water-use efficiency as exemplified by the lowered leaf δ13C under fluctuating soil moisture conditions....... 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...... unresponsive to P but significantly affected by the irrigation treatment. FI plants had the largest SS, followed by DI, and PRD the smallest; and the reverse was the case for SD. Compared to FI and DI, PRD plants had significantly lower values of specific leaf area (SLA) and leaf carbon isotope discrimination...

Stomatal conductance of semi-natural Mediterranean grasslands: Implications for the development of ozone critical levels

International Nuclear Information System (INIS)

Alonso, R.; Bermejo, V.; Sanz, J.; Valls, B.; Elvira, S.; Gimeno, B.S.

2007-01-01

Intra-genus and intra-specific variation and the influence of nitrogen enrichment on net assimilation and stomatal conductance of some annual Trifolium species of Mediterranean dehesa grasslands were assessed under experimental conditions. Also gas exchange rates were compared between some Leguminosae and Poaceae species growing in the field in a dehesa ecosystem in central Spain. The results showed that the previously reported different O 3 sensitivity of some Trifolium species growing in pots does not seem to be related to different maximum g s values. In addition, no clear differences on gas exchange rates could be attributed to Leguminosae and Poaceae families growing in the field, with intra-genus variation being more important than differences found between families. Further studies are needed to increase the database for developing a flux-based approach for setting O 3 critical levels for semi-natural Mediterranean species. - The stomatal conductance model incorporated within the EMEP DO 3 SE deposition module needs to be re-parameterised for Mediterranean semi-natural vegetation

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

Is optimality in stomatal conductance an endogenous process or an emergent property arising from interactions with the environment?

Science.gov (United States)

Resco de Dios, Victor; Gessler, Arthur; Ferrio, Juan Pedro; Bahn, Michael; Milcu, Alexandru; Tissue, David; Voltas, Jordi; Roy, Jacques

2016-04-01

Plants are sessile and poikilothermic organisms that need to respond and adjust promptly to an ever-changing environment. Over a single 24 h period, a plant may experience the same level of variation in radiation as in its entire life-time and, in some climates, the oscillation in day-night temperature and vapor pressure deficit might be of similar magnitude to that experienced across a full year. Plants need to maintain a positive C balance without depleting soil water reserves in the face of such a diverse environment, and feedbacks between assimilation (A) and water losses (E) are thought to have evolved to optimize stomatal conductance (gs). In short, the optimal conductance hypothesis proposes that cross-talks between A and stomatal conductance gs lead to a constant marginal water use (λ) during a day, such that A is maximized and E minimized. The biological mechanism by which biochemical processes would feedback gs remains unknown, but multiple studies have shown empirical support for this hypothesis, leading to its current consideration of theory by many. Here we test whether optimal stomatal conductance is an endogenous property, that is, driven solely by factors internal to the plant, and in the absence of environmental fluctuations. After 5 days of entrainment, where monoculture canopies of bean and of cotton were subjected to the average environmental conditions of an August sunny day in Montpellier (at the CNRS European Ecotron, FR), we kept temperature, relative humidity and photosynthetically active radiation constant for 48 h at the values observed at noon. During this period, we monitored leaf gas exchange continuously every two minutes, and canopy gas exchange every 15 minutes. We observed a periodic oscillation in λ, showing a 24 h period, and consistent with a circadian regulation of water use efficiency. Hourly variations in λ could thus not be explained by the optimal stomatal hypothesis. Moreover, the pattern of variation (of maximal water

Increasing water use efficiency along the C3 to C4 evolutionary pathway: a stomatal optimization perspective.

Science.gov (United States)

Way, Danielle A; Katul, Gabriel G; Manzoni, Stefano; Vico, Giulia

2014-07-01

C4 photosynthesis evolved independently numerous times, probably in response to declining atmospheric CO2 concentrations, but also to high temperatures and aridity, which enhance water losses through transpiration. Here, the environmental factors controlling stomatal behaviour of leaf-level carbon and water exchange were examined across the evolutionary continuum from C3 to C4 photosynthesis at current (400 μmol mol(-1)) and low (280 μmol mol(-1)) atmospheric CO2 conditions. To this aim, a stomatal optimization model was further developed to describe the evolutionary continuum from C3 to C4 species within a unified framework. Data on C3, three categories of C3-C4 intermediates, and C4 Flaveria species were used to parameterize the stomatal model, including parameters for the marginal water use efficiency and the efficiency of the CO2-concentrating mechanism (or C4 pump); these two parameters are interpreted as traits reflecting the stomatal and photosynthetic adjustments during the C3 to C4 transformation. Neither the marginal water use efficiency nor the C4 pump strength changed significantly from C3 to early C3-C4 intermediate stages, but both traits significantly increased between early C3-C4 intermediates and the C4-like intermediates with an operational C4 cycle. At low CO2, net photosynthetic rates showed continuous increases from a C3 state, across the intermediates and towards C4 photosynthesis, but only C4-like intermediates and C4 species (with an operational C4 cycle) had higher water use efficiencies than C3 Flaveria. The results demonstrate that both the marginal water use efficiency and the C4 pump strength increase in C4 Flaveria to improve their photosynthesis and water use efficiency compared with C3 species. These findings emphasize that the advantage of the early intermediate stages is predominantly carbon based, not water related. © The Author 2014. Published by Oxford University Press on behalf of the Society for Experimental Biology.

Estimating variation in stomatal frequency at intra-individual, intra-site, and inter-taxonomic levels in populations of the Leonardoxa africana (Fabaceae) complex over environmental gradients in Cameroon

Science.gov (United States)

Finsinger, Walter; Dos Santos, Thibaut; McKey, Doyle

2013-07-01

Variation of stomatal frequency (stomatal density and stomatal index) includes genetically-based, potentially-adaptive variation, and variation due to phenotypic plasticity, the degree of which may be fundamental to the ability to maintain high water-use efficiency and thus to deal with environmental change. We analysed stomatal frequency and morphology (pore length, pore width) in leaves from several individuals from nine populations of four sub-species of the Leonardoxa africana complex. The dataset represents a hierarchical sampling wherein factors are nested within each level (leaves in individuals, individuals in sites, etc.), allowing estimation of the contribution of different levels to overall variation, using variance-component analysis. SI showed significant variation among sites ("site" is largely confounded with "sub-species"), being highest in the sub-species localized in the highest-elevation site. However, most of the observed variance was accounted for at intra-site and intra-individual levels. This variance could reflect great phenotypic plasticity, presumably in response to highly local variation in micro-environmental conditions.

Photodynamic antimicrobial therapy in the treatment of denture stomatitis

International Nuclear Information System (INIS)

Senna, Andre Machado de

2012-01-01

Denture stomatitis (DS), also called chronic atrophic candidiasis, is the most common oral fungal infection in denture wearers. It has a multifactorial etiology, but the presence of Candida spp. biofilm on the denture is considered the most important factor for the establishment of the DS. This study aimed to evaluate the treatment of DS through the use of photodynamic antimicrobial therapy (PAT), mediated by methylene blue. For this purpose, preclinical studies and clinical trials were performed. Simulators prototypes dentures were made of methyl methacrylate polymer to serve as a basis for biofilm growth of the following species of Candida: C. albicans, C. glabrata, C. dubliniensis, C. krusei, C. tropicalis, C. parapsilosis and C. guilliermondii. Methylene blue solution at a concentration of 450 μg/mL was used as a photosensitizer. The prototypes and biofilms were irradiated with a laser of wavelength of 660 nm, potency of 100 mW, for 80 seconds. For the clinical study, subjects were divided into two groups. The first group received conventional treatment based on the use of antifungal Miconazole. The second group received the treatment by PAT. The preclinical results showed that all species of the genus Candida were susceptible to PAT, with a reduction in colonies that ranged from 2.48 to 3.93 log 10 . Clinical outcomes were evaluated for the reduction of colonies of Candida spp. located in the mucosa and in the prosthesis and relative to the improvement of the clinical aspect of the affected mucosa. Both the conventional therapy and PAT were effective in treating DS. There was no significant statistical difference between PAT and conventional treatment for any of the factors evaluated. Thus, it was concluded that PAT is effective in the treatment of denture stomatitis. (author)

Hazard Characterization of Modified Vaccinia Virus Ankara Vector: What Are the Knowledge Gaps?

Directory of Open Access Journals (Sweden)

Malachy I. Okeke

2017-10-01

Full Text Available Modified vaccinia virus Ankara (MVA is the vector of choice for human and veterinary applications due to its strong safety profile and immunogenicity in vivo. The use of MVA and MVA-vectored vaccines against human and animal diseases must comply with regulatory requirements as they pertain to environmental risk assessment, particularly the characterization of potential adverse effects to humans, animals and the environment. MVA and recombinant MVA are widely believed to pose low or negligible risk to ecosystem health. However, key aspects of MVA biology require further research in order to provide data needed to evaluate the potential risks that may occur due to the use of MVA and MVA-vectored vaccines. The purpose of this paper is to identify knowledge gaps in the biology of MVA and recombinant MVA that are of relevance to its hazard characterization and discuss ongoing and future experiments aimed at providing data necessary to fill in the knowledge gaps. In addition, we presented arguments for the inclusion of uncertainty analysis and experimental investigation of verifiable worst-case scenarios in the environmental risk assessment of MVA and recombinant MVA. These will contribute to improved risk assessment of MVA and recombinant MVA vaccines.

Results of the Study of Helminths-Carrying as a Comorbidity in Children with Herpetic Stomatitis

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E.S. Suerkulov

2016-09-01

Full Text Available The paper analyzes the helminths-carrying in children with herpetic stomatitis according to the data of the department of maxillofacial surgery of the National center of mother and child welfare, and determines the relationship of oral diseases with disorders of various parts of the gastrointestinal tract.

Stomatal conductance, canopy temperature, and leaf area index estimation using remote sensing and OBIA techniques

Science.gov (United States)

S. Panda; D.M. Amatya; G. Hoogenboom

2014-01-01

Remotely sensed images including LANDSAT, SPOT, NAIP orthoimagery, and LiDAR and relevant processing tools can be used to predict plant stomatal conductance (gs), leaf area index (LAI), and canopy temperature, vegetation density, albedo, and soil moisture using vegetation indices like normalized difference vegetation index (NDVI) or soil adjusted...

Modelling of stomatal conductance and ozone deposition flux of Norway Spruce using deposition model

Czech Academy of Sciences Publication Activity Database

Zapletal, M.; Chroust, P.; Večeřa, Zbyněk; Mikuška, Pavel; Cudlín, Pavel; Urban, Otmar; Pokorný, Radek; Czerný, Radek; Janouš, Dalibor; Taufarová, Klára

2009-01-01

Roč. 12, 2-3 (2009), s. 75-81 ISSN 1335-339X R&D Projects: GA MŽP SP/1B7/189/07 Institutional research plan: CEZ:AV0Z60870520; CEZ:AV0Z40310501 Keywords : ozone concentration * ozone deposition * stomatal conductance * deposition velocity * resistance model * tropo-spheric ozone Subject RIV: DG - Athmosphere Sciences, Meteorology

Systemic Virus Infections Differentially Modulate Cell Cycle State and Functionality of Long-Term Hematopoietic Stem Cells In Vivo

Directory of Open Access Journals (Sweden)

Christoph Hirche

2017-06-01

Full Text Available Quiescent long-term hematopoietic stem cells (LT-HSCs are efficiently activated by type I interferon (IFN-I. However, this effect remains poorly investigated in the context of IFN-I-inducing virus infections. Here we report that both vesicular stomatitis virus (VSV and murine cytomegalovirus (MCMV infection induce LT-HSC activation that substantially differs from the effects triggered upon injection of synthetic IFN-I-inducing agents. In both infections, inflammatory responses had to exceed local thresholds within the bone marrow to confer LT-HSC cell cycle entry, and IFN-I receptor triggering was not critical for this activation. After resolution of acute MCMV infection, LT-HSCs returned to phenotypic quiescence. However, non-acute MCMV infection induced a sustained inflammatory milieu within the bone marrow that was associated with long-lasting impairment of LT-HSC function. In conclusion, our results show that systemic virus infections fundamentally affect LT-HSCs and that also non-acute inflammatory stimuli in bone marrow donors can affect the reconstitution potential of bone marrow transplants.

Systemic Virus Infections Differentially Modulate Cell Cycle State and Functionality of Long-Term Hematopoietic Stem Cells In Vivo.

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Hirche, Christoph; Frenz, Theresa; Haas, Simon F; Döring, Marius; Borst, Katharina; Tegtmeyer, Pia-K; Brizic, Ilija; Jordan, Stefan; Keyser, Kirsten; Chhatbar, Chintan; Pronk, Eline; Lin, Shuiping; Messerle, Martin; Jonjic, Stipan; Falk, Christine S; Trumpp, Andreas; Essers, Marieke A G; Kalinke, Ulrich

2017-06-13

Quiescent long-term hematopoietic stem cells (LT-HSCs) are efficiently activated by type I interferon (IFN-I). However, this effect remains poorly investigated in the context of IFN-I-inducing virus infections. Here we report that both vesicular stomatitis virus (VSV) and murine cytomegalovirus (MCMV) infection induce LT-HSC activation that substantially differs from the effects triggered upon injection of synthetic IFN-I-inducing agents. In both infections, inflammatory responses had to exceed local thresholds within the bone marrow to confer LT-HSC cell cycle entry, and IFN-I receptor triggering was not critical for this activation. After resolution of acute MCMV infection, LT-HSCs returned to phenotypic quiescence. However, non-acute MCMV infection induced a sustained inflammatory milieu within the bone marrow that was associated with long-lasting impairment of LT-HSC function. In conclusion, our results show that systemic virus infections fundamentally affect LT-HSCs and that also non-acute inflammatory stimuli in bone marrow donors can affect the reconstitution potential of bone marrow transplants. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

Stomatal behavior in fruits and leaves of the purple passion fruit (Passiflora edulis Sims and fruits and cladodes of the yellow pitaya [Hylocereus megalanthus (K. Schum. ex Vaupel Ralf Bauer

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Camilo Sánchez

2013-04-01

Full Text Available Plants as C3 and CAM react photosynthetically different but both can grow in the same agroecological zone in the tropics. Therefore we studied the behavior of stomatal opening in fruits and leaves of the purple passion fruit and fruits and cladodes of the yellow pitaya was studied under natural growing conditions in Granada and Fusagasuga, Cundinamarca (Colombia. Imprints were made on the surface of leaves, fruits and cladodes using cosmetic enamel impressions. Three cycles were carried out, each cycle took 72 hours, obtaining three different samples every 3 hours; then the impressions were observed by microscope and the opened and closed stomata were counted in each species. In each sampling, data of solar radiation, temperature and relative humidity (RH were measured. The purple passion fruit had the typical behavior of a C3 plant in the leaves as well as the fruits, and a positive correlation between the stomatal aperture and radiation and temperature was found, along with a negative correlation between stomatal aperture and RH. The pitaya showed the typical behavior of a CAM plant with a negative correlation between the stomatal opening and radiation and temperature, as well as a positive correlation between stomatal opening and RH. Radiation, temperature and RH affected the stomatal opening in the fruits and cladodes. Stomatal densities differed greatly between the species and plant organs. In the purple passion fruit, 106.53 stomata per mm² leaf surface were found, but only 12.64 stomata per mm² fruit surface; whereas in the pitaya, 11.28 and 1.43 stomata per mm² were found on the cladodes and fruits, respectively

Inactivation of viruses by pasteurization at 60 °C for 10 h with and without 40% glucose as stabilizer during a new manufacturing process of α2-Macroglobulin from Cohn Fraction IV.

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Huangfu, Chaoji; Ma, Yuyuan; Jia, Junting; Lv, Maomin; Zhu, Fengxuan; Ma, Xiaowei; Zhao, Xiong; Zhang, Jingang

2017-03-01

Pasteurization is regularly used to inactivate viruses for the safety of plasma derivatives. Influence of pasteurization at 60 °C for 10 h on α2-Macroglobulin activity and virus inactivation were studied. With 40% sugar as stabilizers more than 70% α2-Macroglobulin activity was reserved after pasteurization compared with 20% in control. Glucose presented a better activity protection effect than sucrose and maltose. By pasteurization without stabilizer the virus titers of pseudorabies virus, Sindbis virus, porcine parvovirus and encephalomyocarditis virus were reduced more than 5.88 log 10 , 7.50 log 10 , 4.88 log 10 , and 5.63 log 10 respectively within 2 h. By pasteurization with 40% glucose vesicular stomatitis virus was inactivated more than 5.88 log 10 within 1 h. Only 2.71 log 10 reduction was achieved for encephalomyocarditis virus after 10 h. 40% glucose protected α2-M activity and viruses simultaneously from pasteurization. Other viral inactivation methods need to be incorporated to ensure viral safety of this manufacturing process of α2-Macroglobulin. Copyright © 2017 International Alliance for Biological Standardization. Published by Elsevier Ltd. All rights reserved.

Development of a dual-protective live attenuated vaccine against H5N1 and H9N2 avian influenza viruses by modifying the NS1 gene.

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Choi, Eun-hye; Song, Min-Suk; Park, Su-Jin; Pascua, Philippe Noriel Q; Baek, Yun Hee; Kwon, Hyeok-il; Kim, Eun-Ha; Kim, Semi; Jang, Hyung-Kwan; Poo, Haryoung; Kim, Chul-Joong; Choi, Young Ki

2015-07-01

An increasing number of outbreaks of avian influenza H5N1 and H9N2 viruses in poultry have caused serious economic losses and raised concerns for human health due to the risk of zoonotic transmission. However, licensed H5N1 and H9N2 vaccines for animals and humans have not been developed. Thus, to develop a dual H5N1 and H9N2 live-attenuated influenza vaccine (LAIV), the HA and NA genes from a virulent mouse-adapted avian H5N2 (A/WB/Korea/ma81/06) virus and a recently isolated chicken H9N2 (A/CK/Korea/116/06) virus, respectively, were introduced into the A/Puerto Rico/8/34 backbone expressing truncated NS1 proteins (NS1-73, NS1-86, NS1-101, NS1-122) but still possessing a full-length NS gene. Two H5N2/NS1-LAIV viruses (H5N2/NS1-86 and H5N2/NS1-101) were highly attenuated compared with the full-length and remaining H5N2/NS-LAIV viruses in a mouse model. Furthermore, viruses containing NS1 modifications were found to induce more IFN-β activation than viruses with full-length NS1 proteins and were correspondingly attenuated in mice. Intranasal vaccination with a single dose (10(4.0) PFU/ml) of these viruses completely protected mice from a lethal challenge with the homologous A/WB/Korea/ma81/06 (H5N2), heterologous highly pathogenic A/EM/Korea/W149/06 (H5N1), and heterosubtypic highly virulent mouse-adapted H9N2 viruses. This study clearly demonstrates that the modified H5N2/NS1-LAIV viruses attenuated through the introduction of mutations in the NS1 coding region display characteristics that are desirable for live attenuated vaccines and hold potential as vaccine candidates for mammalian hosts.

Sodium Lauryl Sulfate Abrogates Human Immunodeficiency Virus Infectivity by Affecting Viral Attachment

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Bestman-Smith, Julie; Piret, Jocelyne; Désormeaux, André; Tremblay, Michel J.; Omar, Rabeea F.; Bergeron, Michel G.

2001-01-01

The microbicidal activity of sodium lauryl sulfate (SLS) against human immunodeficiency virus type 1 (HIV-1) was studied in cultured cells. Pretreatment of HIV-1NL4-3 with SLS decreased, in a concentration-dependent manner, its infectivity when using 1G5 as target cells. In the absence of a viral pretreatment period or when 1G5 cells were pretreated with SLS, the surfactant-induced inactivation of viral infectivity was less pronounced, especially at concentrations between 375 and 550 μM. SLS had no effect on HIV-1 when the virus was adsorbed to 1G5 cells by a 2-h incubation period. SLS almost completely inhibited the fusion process by decreasing the attachment of HIV-1 to target cells. SLS also inhibited the infectivity of HIV-1-based luciferase reporter viruses pseudotyped with the amphotropic murine leukemia virus envelope (which enters cells in a CD4-, CCR5-, and CXCR4-independent manner), indicating that SLS may inactivate other envelope viruses. In contrast, no effect was seen with vesicular stomatitis virus envelope glycoprotein G (which enters cells through receptor-mediated endocytosis) pretreated with up to 700 μM SLS. SLS also decreased, in a dose-dependent manner, the HIV-1-dependent syncytium formation between 1G5 and J1.1 cells after a 24-h incubation. The reduction of luciferase activity was more pronounced when J1.1 cells (which express HIV-1 proteins on their surface) were pretreated with SLS rather than 1G5 cells. Taken together, our results suggest that SLS could represent a candidate of choice for use in vaginal microbicides to prevent the sexual transmission of HIV and possibly other pathogens causing sexually transmitted diseases. PMID:11451679

Transpiration and stomatal conductance in a young secondary tropical montane forest: contrasts between native trees and invasive understorey shrubs.

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Ghimire, Chandra Prasad; Bruijnzeel, L Adrian; Lubczynski, Maciek W; Zwartendijk, Bob W; Odongo, Vincent Omondi; Ravelona, Maafaka; van Meerveld, H J Ilja

2018-04-21

It has been suggested that vigorous secondary tropical forests can have very high transpiration rates, but sap flow and stomatal conductance dynamics of trees and shrubs in these forests are understudied. In an effort to address this knowledge gap, sap flow (thermal dissipation method, 12 trees) and stomatal conductance (porometry, six trees) were measured for young (5-7 years) Psiadia altissima (DC.) Drake trees, a widely occurring species dominating young regrowth following abandonment of swidden agriculture in upland eastern Madagascar. In addition, stomatal conductance (gs) was determined for three individuals of two locally common invasive shrubs (Lantana camara L. and Rubus moluccanus L.) during three periods with contrasting soil moisture conditions. Values of gs for the three investigated species were significantly higher and more sensitive to climatic conditions during the wet period compared with the dry period. Further, gs of the understorey shrubs was much more sensitive to soil moisture content than that of the trees. Tree transpiration rates (Ec) were relatively stable during the dry season and were only affected somewhat by soil water content at the end of the dry season, suggesting the trees had continued access to soil water despite drying out of the topsoil. The Ec exhibited a plateau-shaped relation with vapour pressure deficit (VPD), which was attributed to stomatal closure at high VPD. Vapour pressure deficit was the major driver of variation in Ec, during both the wet and the dry season. Overall water use of the trees was modest, possibly reflecting low site fertility after three swidden cultivation cycles. The observed contrast in gs response to soil water and climatic conditions for the trees and shrubs underscores the need to take root distributions into account when modelling transpiration from regenerating tropical forests.

Lower responsiveness of canopy evapotranspiration rate than of leaf stomatal conductance to open-air CO2 elevation in rice.

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Shimono, Hiroyuki; Nakamura, Hirofumi; Hasegawa, Toshihiro; Okada, Masumi

2013-08-01

An elevated atmospheric CO2 concentration ([CO2 ]) can reduce stomatal conductance of leaves for most plant species, including rice (Oryza sativa L.). However, few studies have quantified seasonal changes in the effects of elevated [CO2 ] on canopy evapotranspiration, which integrates the response of stomatal conductance of individual leaves with other responses, such as leaf area expansion, changes in leaf surface temperature, and changes in developmental stages, in field conditions. We conducted a field experiment to measure seasonal changes in stomatal conductance of the uppermost leaves and in the evapotranspiration, transpiration, and evaporation rates using a lysimeter method. The study was conducted for flooded rice under open-air CO2 elevation. Stomatal conductance decreased by 27% under elevated [CO2 ], averaged throughout the growing season, and evapotranspiration decreased by an average of 5% during the same period. The decrease in daily evapotranspiration caused by elevated [CO2 ] was more significantly correlated with air temperature and leaf area index (LAI) rather than with other parameters of solar radiation, days after transplanting, vapor-pressure deficit and FAO reference evapotranspiration. This indicates that higher air temperatures, within the range from 16 to 27 °C, and a larger LAI, within the range from 0 to 4 m(2)  m(-2) , can increase the magnitude of the decrease in evapotranspiration rate caused by elevated [CO2 ]. The crop coefficient (i.e. the evapotranspiration rate divided by the FAO reference evapotranspiration rate) was 1.24 at ambient [CO2 ] and 1.17 at elevated [CO2 ]. This study provides the first direct measurement of the effects of elevated [CO2 ] on rice canopy evapotranspiration under open-air conditions using the lysimeter method, and the results will improve future predictions of water use in rice fields. © 2013 John Wiley & Sons Ltd.

Intranasal Administration of Maleic Anhydride-Modified Human Serum Albumin for Pre-Exposure Prophylaxis of Respiratory Syncytial Virus Infection

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Zhiwu Sun

2015-02-01

Full Text Available Respiratory syncytial virus (RSV is the leading cause of pediatric viral respiratory tract infections. Neither vaccine nor effective antiviral therapy is available to prevent and treat RSV infection. Palivizumab, a humanized monoclonal antibody, is the only product approved to prevent serious RSV infection, but its high cost is prohibitive in low-income countries. Here, we aimed to identify an effective, safe, and affordable antiviral agent for pre-exposure prophylaxis (PrEP of RSV infection in children at high risk. We found that maleic anhydride (ML-modified human serum albumin (HSA, designated ML-HSA, exhibited potent antiviral activity against RSV and that the percentages of the modified lysines and arginies in ML- are correlated with such anti-RSV activity. ML-HSA inhibited RSV entry and replication by interacting with viral G protein and blocking RSV attachment to the target cells, while ML-HAS neither bound to F protein, nor inhibited F protein-mediated membrane fusion. Intranasal administration of ML-HSA before RSV infection resulted in significant decrease of the viral titers in the lungs of mice. ML-HSA shows promise for further development into an effective, safe, affordable, and easy-to-use intranasal regimen for pre-exposure prophylaxis of RSV infection in children at high risk in both low- and high-income countries.

A Study of Waste Management within the COL Florence A. Blanchfield Army Community Hospital, Fort Campbell, Kentucky.

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1981-08-01

besnoiti Borna disease virus Bovine infectious petechial fever virus Camel pox virus Ephemeral fever virus Fowl plague virus Goat pox virus Hog...Varicella virus Vole rickettsia Yellow fever virus, 17D vaccine strain 69 Class 3 Alastrun, smallpox, monkeypox, and whitepox, when used in vitro Arbovirus...animal inoculation experiments Vesicular stomatitis virus Yellow fever virus - wild when used in vitro Class 4 Alastrun, smallpox, monkeypox, and

Oronasal and intramuscular vaccination of swine with a modified live porcine parvovirus vaccine: multiplication and transmission of the vaccine virus.

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Paul, P S; Mengeling, W L

1984-12-01

An attenuated strain NADL-2 of porcine parvovirus (PPV) has been used at the 54th cell culture passage as a modified live-virus (MLV) vaccine. The present study was conducted to determine the minimum immunizing dose of MLV, the extent of MLV multiplication in swine tissues, and its transmission from swine administered MLV oronasally or intramuscularly. Immune response to MLV was dose dependent and swine responded to as little as 10(2) median cell-culture infective doses (CCID50). A 10(5) CCID50 of MLV, the largest dose given, induced the best immune response and was used in subsequent experiments. Route of MLV administration also was found to be important. The MLV replicated in tissues of swine after IM inoculation; however, viral antigen in tissues was less, as measured by immunofluorescence, and serum hemagglutination-inhibition titers for PPV were lower in MLV-inoculated swine than we have previously observed in virulent PPV-inoculated swine. In contrast, oronasal inoculation with MLV did not consistently result in infection of pigs; only 5 of 23 swine had virologic and/or serologic evidence of infection. Virus transmission studies indicated that MLV is shed in feces, but shedding occurs later than that in virulent-PPV-inoculated swine and is inconsistent. Delayed transmission of MLV was observed in contact pigs, which were seronegative at 2 weeks, but became seropositive at 4 weeks--indicating that perhaps a virus population capable of infecting pigs by oronasal route was selected by passage through the pig.(ABSTRACT TRUNCATED AT 250 WORDS)

Enhanced protection against Ebola virus mediated by an improved adenovirus-based vaccine.

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Richardson, Jason S; Yao, Michel K; Tran, Kaylie N; Croyle, Maria A; Strong, James E; Feldmann, Heinz; Kobinger, Gary P

2009-01-01

The Ebola virus is transmitted by direct contact with bodily fluids of infected individuals, eliciting death rates as high as 90% among infected humans. Currently, replication defective adenovirus-based Ebola vaccine is being studied in a phase I clinical trial. Another Ebola vaccine, based on an attenuated vesicular stomatitis virus has shown efficacy in post-exposure treatment of nonhuman primates to Ebola infection. In this report, we modified the common recombinant adenovirus serotype 5-based Ebola vaccine expressing the wild-type ZEBOV glycoprotein sequence from a CMV promoter (Ad-CMVZGP). The immune response elicited by this improved expression cassette vector (Ad-CAGoptZGP) and its ability to afford protection against lethal ZEBOV challenge in mice was compared to the standard Ad-CMVZGP vector. Ad-CMVZGP was previously shown to protect mice, guinea pigs and nonhuman primates from an otherwise lethal challenge of Zaire ebolavirus. The antigenic expression cassette of this vector was improved through codon optimization, inclusion of a consensus Kozak sequence and reconfiguration of a CAG promoter (Ad-CAGoptZGP). Expression of GP from Ad-CAGoptZGP was substantially higher than from Ad-CMVZGP. Ad-CAGoptZGP significantly improved T and B cell responses at doses 10 to 100-fold lower than that needed with Ad-CMVZGP. Additionally, Ad-CAGoptZGP afforded full protections in mice against lethal challenge at a dose 100 times lower than the dose required for Ad-CMVZGP. Finally, Ad-CAGoptZGP induced full protection to mice when given 30 minutes post-challenge. We describe an improved adenovirus-based Ebola vaccine capable of affording post-exposure protection against lethal challenge in mice. The molecular modifications of the new improved vaccine also translated in the induction of significantly enhanced immune responses and complete protection at a dose 100 times lower than with the previous generation adenovirus-based Ebola vaccine. Understanding and improving the

Enhanced protection against Ebola virus mediated by an improved adenovirus-based vaccine.

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Jason S Richardson

Full Text Available BACKGROUND: The Ebola virus is transmitted by direct contact with bodily fluids of infected individuals, eliciting death rates as high as 90% among infected humans. Currently, replication defective adenovirus-based Ebola vaccine is being studied in a phase I clinical trial. Another Ebola vaccine, based on an attenuated vesicular stomatitis virus has shown efficacy in post-exposure treatment of nonhuman primates to Ebola infection. In this report, we modified the common recombinant adenovirus serotype 5-based Ebola vaccine expressing the wild-type ZEBOV glycoprotein sequence from a CMV promoter (Ad-CMVZGP. The immune response elicited by this improved expression cassette vector (Ad-CAGoptZGP and its ability to afford protection against lethal ZEBOV challenge in mice was compared to the standard Ad-CMVZGP vector. METHODOLOGY/PRINCIPAL FINDINGS: Ad-CMVZGP was previously shown to protect mice, guinea pigs and nonhuman primates from an otherwise lethal challenge of Zaire ebolavirus. The antigenic expression cassette of this vector was improved through codon optimization, inclusion of a consensus Kozak sequence and reconfiguration of a CAG promoter (Ad-CAGoptZGP. Expression of GP from Ad-CAGoptZGP was substantially higher than from Ad-CMVZGP. Ad-CAGoptZGP significantly improved T and B cell responses at doses 10 to 100-fold lower than that needed with Ad-CMVZGP. Additionally, Ad-CAGoptZGP afforded full protections in mice against lethal challenge at a dose 100 times lower than the dose required for Ad-CMVZGP. Finally, Ad-CAGoptZGP induced full protection to mice when given 30 minutes post-challenge. CONCLUSIONS/SIGNIFICANCE: We describe an improved adenovirus-based Ebola vaccine capable of affording post-exposure protection against lethal challenge in mice. The molecular modifications of the new improved vaccine also translated in the induction of significantly enhanced immune responses and complete protection at a dose 100 times lower than with the

In situ temperature relationships of biochemical and stomatal controls of photosynthesis in four lowland tropical tree species.

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Slot, Martijn; Winter, Klaus

2017-12-01

Net photosynthetic carbon uptake of Panamanian lowland tropical forest species is typically optimal at 30-32 °C. The processes responsible for the decrease in photosynthesis at higher temperatures are not fully understood for tropical trees. We determined temperature responses of maximum rates of RuBP-carboxylation (V CMax ) and RuBP-regeneration (J Max ), stomatal conductance (G s ), and respiration in the light (R Light ) in situ for 4 lowland tropical tree species in Panama. G s had the lowest temperature optimum (T Opt ), similar to that of net photosynthesis, and photosynthesis became increasingly limited by stomatal conductance as temperature increased. J Max peaked at 34-37 °C and V CMax ~2 °C above that, except in the late-successional species Calophyllum longifolium, in which both peaked at ~33 °C. R Light significantly increased with increasing temperature, but simulations with a photosynthesis model indicated that this had only a small effect on net photosynthesis. We found no evidence for Rubisco-activase limitation of photosynthesis. T Opt of V CMax and J Max fell within the observed in situ leaf temperature range, but our study nonetheless suggests that net photosynthesis of tropical trees is more strongly influenced by the indirect effects of high temperature-for example, through elevated vapour pressure deficit and resulting decreases in stomatal conductance-than by direct temperature effects on photosynthetic biochemistry and respiration. © 2017 John Wiley & Sons Ltd.

Rice Stomatal Closure Requires Guard Cell Plasma Membrane ATP-Binding Cassette Transporter RCN1/OsABCG5.

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Matsuda, Shuichi; Takano, Sho; Sato, Moeko; Furukawa, Kaoru; Nagasawa, Hidetaka; Yoshikawa, Shoko; Kasuga, Jun; Tokuji, Yoshihiko; Yazaki, Kazufumi; Nakazono, Mikio; Takamure, Itsuro; Kato, Kiyoaki

2016-03-07

Water stress is one of the major environmental stresses that affect agricultural production worldwide. Water loss from plants occurs primarily through stomatal pores. Here, we report that an Oryza sativa half-size ATP-binding cassette (ABC) subfamily G protein, RCN1/OsABCG5, is involved in stomatal closure mediated by phytohormone abscisic acid (ABA) accumulation in guard cells. We found that the GFP-RCN1/OsABCG5-fusion protein was localized at the plasma membrane in guard cells. The percentage of guard cell pairs containing both ABA and GFP-RCN1/OsABCG5 increased after exogenous ABA treatment, whereas they were co-localized in guard cell pairs regardless of whether exogenous ABA was applied. ABA application resulted in a smaller increase in the percentage of guard cell pairs containing ABA in rcn1 mutant (A684P) and RCN1-RNAi than in wild-type plants. Furthermore, polyethylene glycol (drought stress)-inducible ABA accumulation in guard cells did not occur in rcn1 mutants. Stomata closure mediated by exogenous ABA application was strongly reduced in rcn1 mutants. Finally, rcn1 mutant plants had more rapid water loss from detached leaves than the wild-type plants. These results indicate that in response to drought stress, RCN1/OsABCG5 is involved in accumulation of ABA in guard cells, which is indispensable for stomatal closure. Copyright © 2016 The Author. Published by Elsevier Inc. All rights reserved.

Stability and Bifurcation Analysis of a Modified Epidemic Model for Computer Viruses

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Chuandong Li

2014-01-01

Full Text Available We extend the three-dimensional SIR model to four-dimensional case and then analyze its dynamical behavior including stability and bifurcation. It is shown that the new model makes a significant improvement to the epidemic model for computer viruses, which is more reasonable than the most existing SIR models. Furthermore, we investigate the stability of the possible equilibrium point and the existence of the Hopf bifurcation with respect to the delay. By analyzing the associated characteristic equation, it is found that Hopf bifurcation occurs when the delay passes through a sequence of critical values. An analytical condition for determining the direction, stability, and other properties of bifurcating periodic solutions is obtained by using the normal form theory and center manifold argument. The obtained results may provide a theoretical foundation to understand the spread of computer viruses and then to minimize virus risks.

Rubella virus detection by ELISA method in exposed radiation workers

International Nuclear Information System (INIS)

Wu Jianmei; Zhu Bo; Zhu Youming; Shao Jinhui; Wu Weiping; Han Jinxiang

2005-01-01

Objective: A rapid diagnosis method was developed to detect Rubella virus infection in radiation workers. Methods: Modified ELISA method was used to detect the level of lgG and lgM antibodies in 514 in Jinan district. Results: 90.47% of 514 cases was shown to be resistant against Rubella virus; 6.42% were sensitive type; 0.78% belonged to be reinfected. Conclusion: Detection of Rubella virus in exposed radiation workers was imperative, and vaccine against Rubella virus was also needed to eliminate the infection risk. (authors)

Clinical efficacy of melittin in the treatment of cats infected with the feline immunodeficiency virus.

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Hartmann, Anja D; Wilhelm, Natalie; Erfle, Volker; Hartmann, Katrin

2016-12-05

The bee venom melittin shows an antiviral efficacy against the human immunodeficiency virus in cell culture. It was shown to be non-toxic for cats. Aim of this pilot study was to investigate the clinical efficacy and side-effects of melittin in cats naturally infected with feline immunodeficiency virus (FIV). The study was performed as a prospective, placebo-controlled double-blinded trial. Twenty cats were included, of which 10 cats each were treated with either melittin (500 µg/kg body weight) or phosphate-buffered saline (placebo) subcutaneously twice per week. During the treatment period of 6 weeks, the cats' general health status, determined by the Karnofsky's score, and the severity of clinical signs (conjunctivitis and stomatitis) using a clinical scoring system were evaluated. Haematology, biochemistry profiles, lymphocyte subpopulations, CD4/CD8 ratio, and pterines (biopterine, 7-xanthopterine) as surrogate parameters were also compared. The general health status and the clinical scores for conjunctivitis and stomatitis improved in cats treated with melittin. A statistically significant improvement however could only be detected for conjunctivitis in cats treated with melittin compared to cats treated with placebo which was likely due to different scores between both groups at the beginning. No influence on the lymphocyte subpopulations, CD4/CD8 ratio, and pterine concentrations was observed. No side effects occurred in this study. In the protocol used in the present study, no significant efficacy of melittin could be detected. However, efficacy of melittin, especially if applied in a higher dosage as in the present study or for a longer period, could be evaluated in further studies. Synergistic effects if used in combination with classic antiretroviral drugs could be an interesting future approach.

Real-time reverse transcription polymerase chain reaction method for detection of Canine distemper virus modified live vaccine shedding for differentiation from infection with wild-type strains.

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Wilkes, Rebecca P; Sanchez, Elena; Riley, Matthew C; Kennedy, Melissa A

2014-01-01

Canine distemper virus (CDV) remains a common cause of infectious disease in dogs, particularly in high-density housing situations such as shelters. Vaccination of all dogs against CDV is recommended at the time of admission to animal shelters and many use a modified live virus (MLV) vaccine. From a diagnostic standpoint for dogs with suspected CDV infection, this is problematic because highly sensitive diagnostic real-time reverse transcription polymerase chain reaction (RT-PCR) tests are able to detect MLV virus in clinical samples. Real-time PCR can be used to quantitate amount of virus shedding and can differentiate vaccine strains from wild-type strains when shedding is high. However, differentiation by quantitation is not possible in vaccinated animals during acute infection, when shedding is low and could be mistaken for low level vaccine virus shedding. While there are gel-based RT-PCR assays for differentiation of vaccine strains from field strains based on sequence differences, the sensitivity of these assays is unable to match that of the real-time RT-PCR assay currently used in the authors' laboratory. Therefore, a real-time RT-PCR assay was developed that detects CDV MLV vaccine strains and distinguishes them from wild-type strains based on nucleotide sequence differences, rather than the amount of viral RNA in the sample. The test is highly sensitive, with detection of as few as 5 virus genomic copies (corresponding to 10(-1) TCID(50)). Sequencing of the DNA real-time products also allows phylogenetic differentiation of the wild-type strains. This test will aid diagnosis during outbreaks of CDV in recently vaccinated animals.

Attenuation and immunogenicity of host-range extended modified vaccinia virus Ankara recombinants.

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Melamed, Sharon; Wyatt, Linda S; Kastenmayer, Robin J; Moss, Bernard

2013-09-23

Modified vaccinia virus Ankara (MVA) is being widely investigated as a safe smallpox vaccine and as an expression vector to produce vaccines against other infectious diseases and cancer. MVA was isolated following more than 500 passages in chick embryo fibroblasts and suffered several major deletions and numerous small mutations resulting in replication defects in human and most other mammalian cells as well as severe attenuation of pathogenicity. Due to the host range restriction, primary chick embryo fibroblasts are routinely used for production of MVA-based vaccines. While a replication defect undoubtedly contributes to safety of MVA, it is worth considering whether host range and attenuation are partially separable properties. Marker rescue transfection experiments resulted in the creation of recombinant MVAs with extended mammalian cell host range. Here, we characterize two host-range extended rMVAs and show that they (i) have acquired the ability to stably replicate in Vero cells, which are frequently used as a cell substrate for vaccine manufacture, (ii) are severely attenuated in immunocompetent and immunodeficient mouse strains following intranasal infection, (iii) are more pathogenic than MVA but less pathogenic than the ACAM2000 vaccine strain at high intracranial doses, (iv) do not form lesions upon tail scratch in mice in contrast to ACAM2000 and (v) induce protective humoral and cell-mediated immune responses similar to MVA. The extended host range of rMVAs may be useful for vaccine production. Published by Elsevier Ltd.

Comparison of arabidopsis stomatal density mutants indicates variation in water stress responses and potential epistatic effects

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Shaneka S. Lawson; Paula M. Pijut; Charles H. Michler

2014-01-01

Recent physiological analysis of Arabidopsis stomatal density (SD) mutants indicated that SD was not the major factor controlling aboveground biomass accumulation. Despite the general theory that plants with fewer stomata have limited biomass acquisition capabilities, epf1 and several other Arabidopsis mutants varied significantly in leaf fresh...

Modelling of root ABA synthesis, stomatal conductance, transpiration and potato production under water saving irrigation regimes

DEFF Research Database (Denmark)

Plauborg, Finn; Abrahamsen, Per; Gjettermann, Birgitte

2010-01-01

. Experimental data was compared to simulated results from the new enhanced Daisy model which include modelling 2D soil water flow, abscisic acid (ABA) signalling and its effect on stomatal conductance and hence on transpiration and assimilation, and finally crop yield. The results demonstrated that the enhanced...

Human Immunodeficiency Virus type-1 reverse transcriptase exists as post-translationally modified forms in virions and cells

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Warrilow David

2008-12-01

Full Text Available Abstract Background HIV-1 reverse transcriptase (RT is a heterodimer composed of p66 and p51 subunits and is responsible for reverse transcription of the viral RNA genome into DNA. RT can be post-translationally modified in vitro which may be an important mechanism for regulating RT activity. Here we report detection of different p66 and p51 RT isoforms by 2D gel electrophoresis in virions and infected cells. Results Major isoforms of the p66 and p51 RT subunits were observed, with pI's of 8.44 and 8.31 respectively (p668.44 and p518.31. The same major isoforms were present in virions, virus-infected cell lysates and intracellular reverse transcription complexes (RTCs, and their presence in RTCs suggested that these are likely to be the forms that function in reverse transcription. Several minor RT isoforms were also observed. The observed pIs of the RT isoforms differed from the pI of theoretical unmodified RT (p668.53 and p518.60, suggesting that most of the RT protein in virions and cells is post-translationally modified. The modifications of p668.44 and p518.31 differed from each other indicating selective modification of the different RT subunits. The susceptibility of RT isoforms to phosphatase treatment suggested that some of these modifications were due to phosphorylation. Dephosphorylation, however, had no effect on in vitro RT activity associated with virions, infected cells or RTCs suggesting that the phospho-isoforms do not make a major contribution to RT activity in an in vitro assay. Conclusion The same major isoform of p66 and p51 RT is found in virions, infected cells and RTC's and both of these subunits are post-translationally modified. This post-translational modification of RT may be important for the function of RT inside the cell.

Evaluation of Multiple Mechanistic Hypotheses of Leaf Photosynthesis and Stomatal Conductance against Diurnal and Seasonal Data from Two Contrasting Panamanian Tropical Forests

Science.gov (United States)

Serbin, S.; Walker, A. P.; Wu, J.; Ely, K.; Rogers, A.; Wolfe, B.

2017-12-01

Tropical forests play a key role in regulating the global carbon (C), water, and energy cycles and stores, as well as influence climate through the exchanges of mass and energy with the atmosphere. However, projected changes in temperature and precipitation patterns are expected to impact the tropics and the strength of the tropical C sink, likely resulting in significant climate feedbacks. Moreover, the impact of stronger, longer, and more extensive droughts not well understood. Critical for the accurate modeling of the tropical C and water cycle in Earth System Models (ESMs) is the representation of the coupled photosynthetic and stomatal conductance processes and how these processes are impacted by environmental and other drivers. Moreover, the parameterization and representation of these processes is an important consideration for ESM projections. We use a novel model framework, the Multi-Assumption Architecture and Testbed (MAAT), together with the open-source bioinformatics toolbox, the Predictive Ecosystem Analyzer (PEcAn), to explore the impact of the multiple mechanistic hypotheses of coupled photosynthesis and stomatal conductance as well as the additional uncertainty related to model parameterization. Our goal was to better understand how model choice and parameterization influences diurnal and seasonal modeling of leaf-level photosynthesis and stomatal conductance. We focused on the 2016 ENSO period and starting in February, monthly measurements of diurnal photosynthesis and conductance were made on 7-9 dominant species at the two Smithsonian canopy crane sites. This benchmark dataset was used to test different representations of stomatal conductance and photosynthetic parameterizations with the MAAT model, running within PEcAn. The MAAT model allows for the easy selection of competing hypotheses to test different photosynthetic modeling approaches while PEcAn provides the ability to explore the uncertainties introduced through parameterization. We

Controlled immobilisation of active enzymes on the cowpea mosaic virus capsid

Science.gov (United States)

Aljabali, Alaa A. A.; Barclay, J. Elaine; Steinmetz, Nicole F.; Lomonossoff, George P.; Evans, David J.

2012-08-01

Immobilisation of horseradish peroxidase (HRP) and glucose oxidase (GOX) via covalent attachment of modified enzyme carbohydrate to the exterior of the cowpea mosaic virus (CPMV) capsid gave high retention of enzymatic activity. The number of enzymes bound per virus was determined to be about eleven for HRP and 2-3 for GOX. This illustrates that relatively large biomacromolecules can be readily coupled to the virus surface using simple conjugation strategies. Virus-biomacromolecule hybrids have great potential for uses in catalysis, diagnostic assays or biosensors.Immobilisation of horseradish peroxidase (HRP) and glucose oxidase (GOX) via covalent attachment of modified enzyme carbohydrate to the exterior of the cowpea mosaic virus (CPMV) capsid gave high retention of enzymatic activity. The number of enzymes bound per virus was determined to be about eleven for HRP and 2-3 for GOX. This illustrates that relatively large biomacromolecules can be readily coupled to the virus surface using simple conjugation strategies. Virus-biomacromolecule hybrids have great potential for uses in catalysis, diagnostic assays or biosensors. Electronic supplementary information (ESI) available: Alternative conjugation strategies, agarose gel electrophoresis of CPMV and CPMV-HRP conjugates, UV-vis spectrum of HRP-ADHCPMV, agarose gel electrophoresis of GOX-ADHCPMV particles and corresponding TEM image, calibration curves for HRP-ADHCPMV and GOX-ADHCPMV, DLS data for GOX-ADHCPMV are made available. See DOI: 10.1039/c2nr31485a

Growth performance and stomatal behavior in relation to ecotypic adaptations in cynodon dactylon (L.) pers

International Nuclear Information System (INIS)

Tufail, A.; Ahmad, F.; Hameed, M.; Ahmad, R.

2017-01-01

Evolution has great ecological significance in terms of plant morphological and stomatal characteristics that must have been genetically fixed during the long evolutionary period. Impact of environmental conditions on growth and stomatal features of twelve ecotypes of Cynodon dactylon that were collected from ecologically different habitats in the Punjab, Pakistan were evaluated. The collected ecotypes Derawar Fort-saline desert (DF-SD), Muzaffar garh-River bank (M-RB), Khabbeki Lake-hyper saline (KL-HS), Ucchali Lake-hyper saline (UL-HS), Kalar Kahar Lake-saline (KKL-S), Treemu-saline wetland (T-SW), Sahianwala-saline wetland (S-SW), Sahianwala-hyper saline (S-HS), Pakka Anna-hyper saline (PA-HS), Pakka Anna-reclaimed field (PA-RF), Botanic Garden-non saline (BG-NS) and Gatwala-saline semiarid (G-SSA) were grown in controlled environments at University of Agriculture, Faisalabad till their acclimatization to evaluate genetically fixed characteristics. After 6-month growth in soil, the plants were transferred to half-strength Hoagland's nutrient medium. There was a huge variation in all morphological characteristics recorded during the investigation, which were due to environmental heterogeniety to which these ecotypes were originally adapted. An exclusive feature of the DF-SD ecotypes is the long and numerous roots, and tillering capacity that surpassed all other ecotypes. Leaves per plant were also exceptionally high that may improve the photosymthetic efficiency of the plant. It showed a good potential of overall growth and biomass production. The robust growth was also recorded in the KKL-S ecotypes, and this can be related to the complete dominance of these two ecotypes in their respective habitats. Small stomata were recorded in the three ecotypes (DF-SD, KL-HS and PA-HS), which are of great ecological significance. Stomatal shape, however, is different in different ecotypes, but its contribution towards stress tolerance is still to be investigated. (author)

Influence of the Leader protein coding region of foot-and-mouth disease virus on virus replication

DEFF Research Database (Denmark)

Belsham, Graham

2013-01-01

The foot-and-mouth disease virus (FMDV) Leader (L) protein is produced in two forms, Lab and Lb, differing only at their amino-termini, due to the use of separate initiation codons, usually 84 nt apart. It has been shown previously, and confirmed here, that precise deletion of the Lab coding......, in the context of the virus lacking the Lb coding region, was also tolerated by the virus within BHK cells. However, precise loss of the Lb coding sequence alone blocked FMDV replication in primary bovine thyroid cells. Thus, the requirement for the Leader protein coding sequences is highly dependent...... on the nature and extent of the residual Leader protein sequences and on the host cell system used. FMDVs precisely lacking Lb and with the Lab initiation codon modified may represent safer seed viruses for vaccine production....

A reduced order model to analytically infer atmospheric CO2 concentration from stomatal and climate data

Science.gov (United States)

Konrad, Wilfried; Katul, Gabriel; Roth-Nebelsick, Anita; Grein, Michaela

2017-06-01

To address questions related to the acceleration or deceleration of the global hydrological cycle or links between the carbon and water cycles over land, reliable data for past climatic conditions based on proxies are required. In particular, the reconstruction of palaeoatmospheric CO2 content (Ca) is needed to assist the separation of natural from anthropogenic Ca variability and to explore phase relations between Ca and air temperature Ta time series. Both Ta and Ca are needed to fingerprint anthropogenic signatures in vapor pressure deficit, a major driver used to explain acceleration or deceleration phases in the global hydrological cycle. Current approaches to Ca reconstruction rely on a robust inverse correlation between measured stomatal density in leaves (ν) of many plant taxa and Ca. There are two methods that exploit this correlation: The first uses calibration curves obtained from extant species assumed to represent the fossil taxa, thereby restricting the suitable taxa to those existing today. The second is a hybrid eco-hydrological/physiological approach that determines Ca with the aid of systems of equations based on quasi-instantaneous leaf-gas exchange theories and fossil stomatal data collected along with other measured leaf anatomical traits and parameters. In this contribution, a reduced order model (ROM) is proposed that derives Ca from a single equation incorporating the aforementioned stomatal data, basic climate (e.g. temperature), estimated biochemical parameters of assimilation and isotope data. The usage of the ROM is then illustrated by applying it to isotopic and anatomical measurements from three extant species. The ROM derivation is based on a balance between the biochemical demand and atmospheric supply of CO2 that leads to an explicit expression linking stomatal conductance to internal CO2 concentration (Ci) and Ca. The resulting expression of stomatal conductance from the carbon economy of the leaf is then equated to another

ROS signaling and stomatal movement in plant responses to drought stress and pathogen attack.

Science.gov (United States)

Qi, Junsheng; Song, Chun-Peng; Wang, Baoshan; Zhou, Jianmin; Kangasjärvi, Jaakko; Zhu, Jian-Kang; Gong, Zhizhong

2018-04-16

Stomata, the pores formed by a pair of guard cells, are the main gateways for water transpiration and photosynthetic CO 2 exchange, as well as pathogen invasion in land plants. Guard cell movement is regulated by a combination of environmental factors including water status, light, CO 2 levels and pathogen attack, as well as endogenous signals such as abscisic acid and apoplastic reactive oxygen species (ROS). Under abiotic and biotic stress conditions, extracellular ROS are mainly produced by plasma membrane-localized NADPH oxidases, whereas intracellular ROS are produced in multiple organelles. These ROS form a sophisticated cellular signaling network, with the accumulation of apoplastic ROS an early hallmark of stomatal movement. Here, we review recent progress in understanding the molecular mechanisms of the ROS signaling network, primarily during drought stress and pathogen attack. We summarize the roles of apoplastic ROS in regulating stomatal movement, ABA and CO 2 signaling, and immunity responses. Finally, we discuss ROS accumulation and communication between organelles and cells. This information provides a conceptual framework for understanding how ROS signaling is integrated with various signaling pathways during plant responses to abiotic and biotic stress stimuli. This article is protected by copyright. All rights reserved.

Seawater spray injury to Quercus acutissima leaves: crystal deposition, stomatal clogging, and chloroplast degeneration.

Science.gov (United States)

Kim, Ki Woo; Koo, Kyosang; Kim, Pan-Gi

2011-05-01

Effects of seawater spray on leaf structure were investigated in Quercus acutissima by electron microscopy and X-ray microanalysis. Two-year-old seedlings of Q. acutissima were sprayed with seawater and kept in a greenhouse maintained at 25°C. The most recognizable symptoms of seawater-sprayed seedlings included leaf necrosis, crystal deposition, stomatal clogging, and chloroplast degeneration. Field emission scanning electron microscopy revealed that the leaf surface was covered with additional layers of remnants of seawater spray. Composed of sodium and chloride, cube-shaped crystals (halite) were prevalently found on trichomes and epidermis, and formed aggregates. Meanwhile, wedge-shaped crystals were deposited on epidermis and consisted of calcium and sulfur. As a result of stomatal clogging by crystal deposition on the abaxial surface, it was conceivable that plant respiration became severely hampered. Transmission electron microscopy showed degenerated cytoplasm of seawater-sprayed leaves. It was common to observe severe plasmolysis and disrupted chloroplasts with a reduced number of thylakoids in grana. These results indicate that foliar applications of seawater were sufficient to induce necrosis of Q. acutissima seedlings as an abiotic disturbance factor. Copyright © 2010 Wiley-Liss, Inc.

Guard cell zeaxanthin tracks photosynthetically active radiation and stomatal apertures in Vicia faba leaves

International Nuclear Information System (INIS)

Srivastava, A.; Zeiger, E.

1995-01-01

Zeaxanthin, antheraxanthin and violaxanthin concentrations in guard cells from sonicated abaxial epidermal peels of Vicia faba were measured from dawn to dusk, and compared with concentrations in mesophyll tissue of the same leaves. Measured changes in guard cell zeaxanthin and violaxanthin concentrations indicate that guard cells operate the xanthophyll cycle throughout the day. Mesophyll tissue had no detectable zeaxanthin at dawn, whereas guard cells had 30–50 mmol mol −1 chlorophyll a+b. On a chlorophyll basis, maximal zeaxanthin levels were 3–4 fold higher in guard cells than in mesophyll cells. Zeaxanthin concentrations tracked levels of photosynthetically active radiation (PAR) in both mesophyll and guard cells. In the mesophyll, most of the zeaxanthin changes occurred in mid-morning and mid-afternoon. In guard cells, zeaxanthin concentrations changed nearly linearly with PAR in the early morning and late afternoon, and closely tracked PAR levels throughout the day. Guard cell zeaxanthin concentrations were also closely correlated with stomatal apertures. The close relationship between zeaxanthin concentrations and PAR levels in guard cells indicates that zeaxanthin is well suited to function as a molecular photosensor in stomatal movements. (author)

OsPRX2 contributes to stomatal closure and improves potassium deficiency tolerance in rice.

Science.gov (United States)

Mao, Xiaohui; Zheng, Yanmei; Xiao, Kaizhuan; Wei, Yidong; Zhu, Yongsheng; Cai, Qiuhua; Chen, Liping; Xie, Huaan; Zhang, Jianfu

2018-01-01

Peroxiredoxins (Prxs) which are thiol-based peroxidases have been implicated in the toxic reduction and intracellular concentration regulation of hydrogen peroxide. In Arabidopsis thaliana At2-CysPrxB (At5g06290) has been demonstrated to be essential in maintaining the water-water cycle for proper H 2 O 2 scavenging. Although the mechanisms of 2-Cys Prxs have been extensively studied in Arabidopsis thaliana, the function of 2-Cys Prxs in rice is unclear. In this study, a rice homologue gene of At2-CysPrxB, OsPRX2 was investigated aiming to characterize the effect of 2-Cys Prxs on the K + -deficiency tolerance in rice. We found that OsPRX2 was localized in the chloroplast. Overexpressed OsPRX2 causes the stomatal closing and K + -deficiency tolerance increasing, while knockout of OsPRX2 lead to serious defects in leaves phenotype and the stomatal opening under the K + -deficiency tolerance. Detection of K + accumulation, antioxidant activity of transgenic plants under the starvation of potassium, further confirmed that OsPRX2 is a potential target for engineering plants with improved potassium deficiency tolerance. Copyright © 2017 Elsevier Inc. All rights reserved.

Bovine herpes virus infections in cattle.

Science.gov (United States)

Nandi, S; Kumar, Manoj; Manohar, M; Chauhan, R S

2009-06-01

Bovine herpes virus 1 (BHV-1) is primarily associated with clinical syndromes such as rhinotracheitis, pustular vulvovaginitis and balanoposthitis, abortion, infertility, conjunctivitis and encephalitis in bovine species. The main sources of infection are the nasal exudates and the respiratory droplets, genital secretions, semen, fetal fluids and tissues. The BHV-1 virus can become latent following a primary infection with a field isolate or vaccination with an attenuated strain. The viral genomic DNA has been demonstrated in the sensory ganglia of the trigeminal nerve in infectious bovine rhinotracheitis (IBR) and in sacral spinal ganglia in pustular vulvovaginitis and balanoposthitis cases. BHV-1 infections can be diagnosed by detection of virus or virus components and antibody by serological tests or by detection of genomic DNA by polymerase chain reaction (PCR), nucleic acid hybridization and sequencing. Inactivated vaccines and modified live virus vaccines are used for prevention of BHV-1 infections in cattle; subunit vaccines and marker vaccines are under investigation.

Aquaporins facilitate hydrogen peroxide entry into guard cells to mediate ABA- and pathogen-triggered stomatal closure.

Science.gov (United States)

Rodrigues, Olivier; Reshetnyak, Ganna; Grondin, Alexandre; Saijo, Yusuke; Leonhardt, Nathalie; Maurel, Christophe; Verdoucq, Lionel

2017-08-22

Stomatal movements are crucial for the control of plant water status and protection against pathogens. Assays on epidermal peels revealed that, similar to abscisic acid (ABA), pathogen-associated molecular pattern (PAMP) flg22 requires the At PIP2;1 aquaporin to induce stomatal closure. Flg22 also induced an increase in osmotic water permeability ( P f ) of guard cell protoplasts through activation of At PIP2;1. The use of HyPer, a genetic probe for intracellular hydrogen peroxide (H 2 O 2 ), revealed that both ABA and flg22 triggered an accumulation of H 2 O 2 in wild-type but not pip2;1 guard cells. Pretreatment of guard cells with flg22 or ABA facilitated the influx of exogenous H 2 O 2 Brassinosteroid insensitive 1-associated receptor kinase 1 (BAK1) and open stomata 1 (OST1)/Snf1-related protein kinase 2.6 (SnRK2.6) were both necessary to flg22-induced P f and both phosphorylated At PIP2;1 on Ser121 in vitro. Accumulation of H 2 O 2 and stomatal closure as induced by flg22 was restored in pip2;1 guard cells by a phosphomimetic form (Ser121Asp) but not by a phosphodeficient form (Ser121Ala) of At PIP2;1. We propose a mechanism whereby phosphorylation of At PIP2;1 Ser121 by BAK1 and/or OST1 is triggered in response to flg22 to activate its water and H 2 O 2 transport activities. This work establishes a signaling role of plasma membrane aquaporins in guard cells and potentially in other cellular context involving H 2 O 2 signaling.

Wolbachia wStri Blocks Zika Virus Growth at Two Independent Stages of Viral Replication.

Science.gov (United States)

Schultz, M J; Tan, A L; Gray, C N; Isern, S; Michael, S F; Frydman, H M; Connor, J H

2018-05-22

Mosquito-transmitted viruses are spread globally and present a great risk to human health. Among the many approaches investigated to limit the diseases caused by these viruses are attempts to make mosquitos resistant to virus infection. Coinfection of mosquitos with the bacterium Wolbachia pipientis from supergroup A is a recent strategy employed to reduce the capacity for major vectors in the Aedes mosquito genus to transmit viruses, including dengue virus (DENV), Chikungunya virus (CHIKV), and Zika virus (ZIKV). Recently, a supergroup B Wolbachia w Stri, isolated from Laodelphax striatellus , was shown to inhibit multiple lineages of ZIKV in Aedes albopictus cells. Here, we show that w Stri blocks the growth of positive-sense RNA viruses DENV, CHIKV, ZIKV, and yellow fever virus by greater than 99.9%. w Stri presence did not affect the growth of the negative-sense RNA viruses LaCrosse virus or vesicular stomatitis virus. Investigation of the stages of the ZIKV life cycle inhibited by w Stri identified two distinct blocks in viral replication. We found a reduction of ZIKV entry into w Stri-infected cells. This was partially rescued by the addition of a cholesterol-lipid supplement. Independent of entry, transfected viral genome was unable to replicate in Wolbachia -infected cells. RNA transfection and metabolic labeling studies suggested that this replication defect is at the level of RNA translation, where we saw a 66% reduction in mosquito protein synthesis in w Stri-infected cells. This study's findings increase the potential for application of w Stri to block additional arboviruses and also identify specific blocks in viral infection caused by Wolbachia coinfection. IMPORTANCE Dengue, Zika, and yellow fever viruses are mosquito-transmitted diseases that have spread throughout the world, causing millions of infections and thousands of deaths each year. Existing programs that seek to contain these diseases through elimination of the mosquito population have so

The effect of vapour pressure deficit on stomatal conductance, sap pH and leaf-specific hydraulic conductance in Eucalyptus globulus clones grown under two watering regimes.

Science.gov (United States)

Hernandez, Maria Jose; Montes, Fernando; Ruiz, Federico; Lopez, Gustavo; Pita, Pilar

2016-05-01

Stomatal conductance has long been considered of key interest in the study of plant adaptation to water stress. The expected increase in extreme meteorological events under a climate change scenario may compromise survival in Eucalyptus globulus plantations established in south-western Spain. We investigated to what extent changes in stomatal conductance in response to high vapour pressure deficits and water shortage are mediated by hydraulic and chemical signals in greenhouse-grown E. globulus clones. Rooted cuttings were grown in pots and submitted to two watering regimes. Stomatal conductance, shoot water potential, sap pH and hydraulic conductance were measured consecutively in each plant over 4 weeks under vapour pressure deficits ranging 0·42 to 2·25 kPa. Evapotranspiration, growth in leaf area and shoot biomass were also determined. There was a significant effect of both clone and watering regime in stomatal conductance and leaf-specific hydraulic conductance, but not in sap pH. Sap pH decreased as water potential and stomatal conductance decreased under increasing vapour pressure deficit. There was no significant relationship between stomatal conductance and leaf-specific hydraulic conductance. Stomata closure precluded shoot water potential from falling below -1·8 MPa. The percentage loss of hydraulic conductance ranged from 40 to 85 %. The highest and lowest leaf-specific hydraulic conductances were measured in clones from the same half-sib families. Water shortage reduced growth and evapotranspiration, decreases in evapotranspiration ranging from 14 to 32 % in the five clones tested. Changes in sap pH seemed to be a response to changes in atmospheric conditions rather than soil water in the species. Stomata closed after a considerable amount of hydraulic conductance was lost, although intraspecific differences in leaf-specific hydraulic conductance suggest the possibility of selection for improved productivity under water-limiting conditions

Evidence That Drought-Induced Stomatal Closure Is Not an Important Constraint on White Spruce Performance Near the Arctic Treeline in Alaska

Science.gov (United States)

Sullivan, P.; Brownlee, A.; Ellison, S.; Sveinbjornsson, B.

2014-12-01

Tree cores collected from trees growing at high latitudes have long been used to reconstruct past climates, because of close positive correlations between temperature and tree growth. However, in recent decades and at many sites, these relationships have deteriorated and have even become negative in some instances. The observation of declining tree growth in response to rising temperature has prompted many investigators to suggest that high latitude trees may be increasingly exhibiting drought-induced stomatal closure. In the Brooks Range of northern Alaska, the observation of low and declining growth of white spruce is more prevalent in the central and eastern parts of the range, where precipitation is lower, providing superficial support for the drought stress hypothesis. In this study, we investigated the occurrence of white spruce drought-induced stomatal closure in four watersheds along a west to east gradient near the Arctic treeline in the Brooks Range. We obtained a historical perspective on tree growth and water relations by collecting increment cores for analysis of ring widths and carbon isotopes in tree-ring alpha-cellulose. Meanwhile, we made detailed assessments of contemporary water relations at the scales of the whole canopy and the needle. All of our data indicate that drought-induced stomatal closure is probably not responsible for low and declining growth in the central and eastern Brooks Range. Carbon isotope discrimination has generally increased over the past century and our calculations indicate that needle inter-cellular CO2 concentration is much greater now than it was in the early 1900's. Measurements of needle gas exchange are consistent with the tree core record, in the sense that instances of low photosynthesis at our sites are not coincident with similarly low stomatal conductance and low inter-cellular CO2 concentration. Finally, hourly measurements of xylem sap flow indicate that trees at our study sites are able to maintain near

Effect of rutin on virus inactivation by AMT in combination with ultraviolet-A irradiation in platelet concentrates

Energy Technology Data Exchange (ETDEWEB)

Yamada, Yoshiko; Abe, Hideki; Ikebuchi, Kenji; Sekiguchi, Sadayoshi [Hokkaido Red Cross Blood Center, Sapporo (Japan)

1999-10-01

Treatment with psoralens and ultraviolet-A (UVA) irradiation have been found to be effective for virus sterilization of platelet concentrates (PCs). We report here a virus inactivation method using a combination of psoralen derivative 4'-aminomethyl-4,5', 8-trimethylpsoralen (AMT) and UVA irradiation (AMT/UVA). Further, we also investigated the effect of rutin, a radical scavenger, on the inactivation of vesicular stomatitis virus (VSV) as a model virus administered in PCs and platelet functions were investigated. Spiked VSV (about 5log{sub 10}) in PCs was inactivated by a combination of AMT (50 {mu}g/ml) and 5.2 J/cm{sup 2} UVA irradiation in the absence of rutin. To obtain equivalent levels of VSV kill in the presence of 0.35 mM rutin, treatment with 13.0 J/cm{sup 2} of UVA irradiation with AMT was performed. When PCs were treated under each condition in which 5log{sub 10} VSV was inactivated by AMT/UVA with or without rutin, platelet aggregation function was maintained for more than 80% of untreated platelets. These findings indicate that the presence of rutin during AMT/UVA treatment conferred no beneficial effect. In addition, overnight storage of PCs with AMT induced 40% loss of platelet aggregation in response to 10{mu}M ADP. The findings suggest that UVA irradiation is required immediately after the addition of AMT. (author)

Seasonal ozone uptake by a warm-temperate mixed deciduous and evergreen broadleaf forest in western Japan estimated by the Penman–Monteith approach combined with a photosynthesis-dependent stomatal model

International Nuclear Information System (INIS)

Kitao, Mitsutoshi; Komatsu, Masabumi; Hoshika, Yasutomo; Yazaki, Kenichi; Yoshimura, Kenichi; Fujii, Saori; Miyama, Takafumi; Kominami, Yuji

2014-01-01

Canopy-level stomatal conductance over a warm-temperate mixed deciduous and evergreen broadleaf forest in Japan was estimated by the Penman–Monteith approach, as compensated by a semi-empirical photosynthesis-dependent stomatal model, where photosynthesis, relative humidity, and CO 2 concentration were assumed to regulate stomatal conductance. This approach, using eddy covariance data and routine meteorological observations at a flux tower site, permits the continuous estimation of canopy-level O 3 uptake, even when the Penman–Monteith approach is unavailable (i.e. in case of direct evaporation from soil or wet leaves). Distortion was observed between the AOT40 exposure index and O 3 uptake through stomata, as AOT40 peaked in April, but with O 3 uptake occurring in July. Thus, leaf pre-maturation in the predominant deciduous broadleaf tree species (Quercus serrata) might suppress O 3 uptake in springtime, even when the highest O 3 concentrations were observed. -- Highlights: • We estimate canopy-level O 3 uptake in a warm-temperate mixed forest in Japan. • The Penman–Monteith approach is compensated by a photosynthesis-dependent model. • Stomatal conductance can be estimated, even in a partly-opened or wet canopy. • The estimated O 3 dose peaks in summer though O 3 exposure peaks in spring. -- Estimation of seasonal O 3 uptake over a mixed-temperate forest compensated by a photosynthesis-dependent stomatal model

Drought increases cowpea (Vigna unguiculata [L.] Walp.) susceptibility to cowpea severe mosaic virus (CPSMV) at early stage of infection.

Science.gov (United States)

Silva, Rodolpho G G; Vasconcelos, Ilka M; Martins, Thiago F; Varela, Anna L N; Souza, Pedro F N; Lobo, Ana K M; Silva, Fredy D A; Silveira, Joaquim A G; Oliveira, Jose T A

2016-12-01

The physiological and biochemical responses of a drought tolerant, virus-susceptible cowpea genotype exposed to drought stress (D), infected by Cowpea severe mosaic virus (CPSMV) (V), and to these two combined stresses (DV), at 2 and 6 days post viral inoculation (DPI), were evaluated. Gas exchange parameters (net photosynthesis, transpiration rate, stomatal conductance, and internal CO 2 partial pressure) were reduced in D and DV at 2 and 6 DPI compared to control plants (C). Photosynthesis was reduced by stomatal and biochemical limitations. Water use efficiency increased at 2 DPI in D, DV, and V, but at 6 DPI only in D and DV compared to C. Photochemical parameters (effective quantum efficiency of photosystem II and electron transport rate) decreased in D and DV compared to C, especially at 6 DPI. The potential quantum efficiency of photosystem II did not change, indicating reversible photoinhibition of photosystem II. In DV, catalase decreased at 2 and 6 DPI, ascorbate peroxidase increased at 2 DPI, but decreased at 6 DPI. Hydrogen peroxide increased at 2 and 6 DPI. Peroxidase increased at 6 DPI and chitinase at 2 and 6 DPI. β-1,3-glucanase decreased in DV at 6 DPI compared to V. Drought increased cowpea susceptibility to CPSMV at 2 DPI, as verified by RT-PCR. However, at 6 DPI, the cowpea plants overcome this effect. Likewise, CPSMV increased the negative effects of drought at 2 DPI, but not at 6 DPI. It was concluded that the responses to combined stresses are not additive and cannot be extrapolated from the study of individual stresses. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

Progress toward an enhanced vaccine: Eight marked attenuated viruses to porcine reproductive and respiratory disease virus.

Science.gov (United States)

Spear, Allyn; Wang, Feng-Xue; Kappes, Matthew A; Das, Phani B; Faaberg, Kay S

2018-03-01

Recombinant viruses of strain Ingelvac® PRRS porcine reproductive and respiratory syndrome virus (PRRSV) modified live virus vaccine were produced with two individual small in-frame deletions in nonstructural protein 2 (nsp2; Δ23 and Δ87) and also the same deletions supplanted with foreign tags (Δ23-V5, Δ23-FLAG, Δ23-S, Δ87-V5, Δ87-FLAG, Δ87-S). The viruses, but one (Δ87-FLAG), were stable for 10 passages and showed minimal effects on in vitro growth. Northern hybridization showed that the Δ23-tagged probe detected intracellular viral genome RNA as well as shorter RNAs that may represent heteroclite species, while the Δ87-tagged probe detected predominantly only genome length RNAs. When the tagged viruses were used to probe nsp2 protein in infected cells, perinuclear localization similar to native nsp2 was seen. Dual infection of Δ23-S and Δ87-S viruses allowed some discrimination of individual tagged nsp2 protein, facilitating future research. The mutants could potentially also be used to differentiate infected from vaccinated animals. Published by Elsevier Inc.

PFAPA (Periodic fever, aphtous stomatitis, pharingitis, cervical adenitis) or Marshall’s syndrome in children

OpenAIRE

N N Kuzmina; G R Movsisyan

2005-01-01

PFAPA (periodic fever, aphtous stomatitis, pharingitis, cervical adenitis) or Marshall’s syndrome is one of the rare periodic fever conditions appearing in children. Its cause is unknown. This syndrome may continue for several years. During interictal period the child is quite well, grows and develops normally. The disease should be differentiated from Behcet’s disease, cyclic neutropenia, familial Mediterranean fever, familial Ireland fever, hyperimmunoglobulinemia D syndrome, systemic juven...

Dynamics of spatial heterogeneity of stomatal closure in Tradescantia virginiana altered by growth at high relative air humidity

NARCIS (Netherlands)

Rezaei Nejad, A.; Harbinson, J.; Meeteren, van U.

2006-01-01

The spatial heterogeneity of stomatal closure in response to rapid desiccation of excised well-watered Tradescantia virginiana leaves grown at moderate (55%) or high (90%) relative air humidity (RH) was studied using a chlorophyll fluorescence imaging system under non-photorespiratory conditions.

Fatal vaccine-induced canine distemper virus infection in black-footed ferrets

Science.gov (United States)

Carpenter, J.W.; Appel, M.J.G.; Erickson, R.C.; Novilla, M.N.

1976-01-01

Four black-footed ferrets that were live-trapped in South Dakota and transported to the Patuxent Wildlife Research Center died within 21 days after vaccination with modified live canine distemper virus. Immunofluorescence, European ferret inoculation, virus isolation attempts, and serum-neutralization tests indicated insufficient attenuation of the vaccine for this species.

Adaxial/abaxial specification in the regulation of photosynthesis and stomatal opening with respect to light orientation and growth with CO2 enrichment in the C4 species Paspalum dilatatum.

Science.gov (United States)

Soares, Ana Sofia; Driscoll, Simon P; Olmos, Enrique; Harbinson, Jeremy; Arrabaça, Maria Celeste; Foyer, Christine H

2008-01-01

Whole-plant morphology, leaf structure and composition were studied together with the effects of light orientation on the dorso-ventral regulation of photosynthesis and stomatal conductance in Paspalum dilatatum cv. Raki plants grown for 6 wk at either 350 or 700 microl l(-1) CO(2). Plant biomass was doubled as a result of growth at high CO(2) and the shoot:root ratio was decreased. Stomatal density was increased in the leaves of the high CO(2)-grown plants, which had greater numbers of smaller stomata and more epidermal cells on the abaxial surface. An asymmetric surface-specific regulation of photosynthesis and stomatal conductance was observed with respect to light orientation. This was not caused by dorso-ventral variations in leaf structure, the distribution of phosphoenolpyruvate carboxylase (PEPC) and ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) proteins or light absorptance, transmittance or reflectance. Adaxial/abaxial specification in the regulation of photosynthesis results from differential sensitivity of stomatal opening to light orientation and fixed gradients of enzyme activation across the leaf.

Prevention of everolimus-related stomatitis in women with hormone receptor-positive, HER2-negative metastatic breast cancer using dexamethasone mouthwash (SWISH): a single-arm, phase 2 trial.

Science.gov (United States)

Rugo, Hope S; Seneviratne, Lasika; Beck, J Thaddeus; Glaspy, John A; Peguero, Julio A; Pluard, Timothy J; Dhillon, Navneet; Hwang, Leon Christopher; Nangia, Chaitali; Mayer, Ingrid A; Meiller, Timothy F; Chambers, Mark S; Sweetman, Robert W; Sabo, J Randy; Litton, Jennifer K

2017-05-01

Stomatitis is a class effect associated with the inhibition of mTOR and is associated with everolimus therapy for breast cancer. Topical steroids might reduce stomatitis incidence and severity, and the need for dose reductions and interruptions of everolimus. Anecdotal use of topical steroid oral prophylaxis has been reported in patients with breast cancer. We aimed to assess dexamethasone-based mouthwash for prevention of stomatitis in patients with breast cancer. This US-based, multicentre, single-arm, phase 2 prevention study enrolled women aged 18 years and older with postmenopausal status who had histologically or cytologically confirmed metastatic hormone receptor-positive, HER2-negative breast cancer. Beginning on day 1 of cycle 1, patients received everolimus 10 mg plus exemestane 25 mg daily, with 10 mL of alcohol-free dexamethasone 0·5 mg per 5 mL oral solution (swish for 2 min and spit, four times daily for 8 weeks). After 8 weeks, dexamethasone mouthwash could be continued for up to eight additional weeks at the discretion of the clinician and patient. The primary endpoint was incidence of grade 2 or worse stomatitis by 8 weeks assessed in the full analysis set (patients who received at least one dose of everolimus and exemestane and at least one confirmed dose of dexamethasone mouthwash) versus historical controls from the BOLERO-2 trial (everolimus and exemestane treatment in patients with hormone receptor-positive advanced breast cancer who were not given dexamethasone mouthwash for prevention of stomatitis). This trial is registered at ClinicalTrials.gov, number NCT02069093. Between May 28, 2014, and Oct 8, 2015, we enrolled 92 women; 85 were evaluable for efficacy. By 8 weeks, the incidence of grade 2 or worse stomatitis was two (2%) of 85 patients (95% CI 0·29-8·24), versus 159 (33%) of 482 patients (95% CI 28·8-37·4) for the duration of the BOLERO-2 study. Overall, 83 (90%) of 92 patients had at least one adverse event. The most frequently