text:
Mini Review ARTICLE
Plant cell wall integrity maintenance as an essential component of biotic stress response mechanisms
- Division of Cell and Molecular Biology, Department of Life Sciences, Imperial College London, London, UK
Plant cell walls provide structural support during development and represent together with the cuticle the first line of defense against biotic and abiotic stress. In recent years, evidence has accumulated that a dedicated plant cell wall integrity (CWI) maintenance mechanism exists. This mechanism monitors and maintains functional integrity of the cell wall during different biological processes. The available data suggest that it may represent a component of the stress response mechanisms underlying biotic and abiotic stress responses, which has not been identified previously as a distinct mechanism. Here I will review the available evidence regarding the mode of action of the CWI maintenance mechanism and discuss its role in the context of biotic plant stress response mechanisms.
Plant cells need to maintain the functional integrity of their walls during cell morphogenesis and exposure to biotic/abiotic stress. The available evidence suggests that a dedicated plant cell wall integrity (CWI) maintenance mechanism exists (Wolf et al., 2011). While our understanding of the mechanisms regulating stress responses and morphogenesis has increased significantly, our knowledge regarding the processes maintaining CWI is still limited. In the last years several reviews have been published on the plant CWI maintenance mechanism illustrating the increased interest in this area (Humphrey et al., 2007; Hematy et al., 2009; Ringli, 2010; Seifert and Blaukopf, 2010). A recently published review focuses on CWI maintenance during plant cell wall morphogenesis (Wolf et al., 2011). Similarities between the yeast and plant CWI maintenance mechanisms have also been reviewed (Hamann and Denness, 2011). Therefore, the available knowledge regarding CWI maintenance during plant development and in yeast will be covered here only briefly to provide a conceptual framework regarding cellular processes involved and to illustrate the degree of functional conservation between species. This review will focus on recent developments regarding the role of CWI maintenance during biotic stress responses. It will discuss how CWI maintenance could have a previously unrecognized role in the perception of and response to biotic stress.
Similarities and Differences Between Yeast and Plant Cell Wall Integrity Maintenance Mechanisms
While both plant and yeast cells are enveloped by cell walls, certain important differences exist that affect the biological role and function of the plant CWI maintenance mechanism. Yeast unlike plant cells, do not have to deal with biotic stress. In addition, plant cell walls are structurally and chemically more complex than the yeast cell wall. This means that in plant cells the sheer number of cell wall-related signaling events during development and plant– environment interaction could disguise the activity of a dedicated plant CWI maintenance mechanism.
The yeast CWI monitoring and maintenance network is quite complex, providing an indication of the possible complexity of the plant CWI maintenance network. By combining inputs from a turgor pressure sensor (SLN1), mechano-perception (MID1/CCH1), and dedicated cell wall damage (CWD) sensors (WSC1, 2, 3, MID2, MTL1) the yeast CWI maintenance network generates signals that permit highly specific responses to any challenge that impairs the functional integrity of the yeast cell wall. The available phenotypic and genetic data also implicate turgor pressure, mechano-perception, and CWD detection in plant CWI maintenance (Hamann et al., 2009; Denness et al., 2011). Interestingly, ARABIDOPSIS HISTIDINE KINASE1 (AHK1) and CYTOKININ RECEPTOR1/ARABIDOPSIS HISTIDINE KINASE4 (CRE1/AHK4) can at least partially rescue a yeast strain with a loss of function allele in SLN1 (Urao et al., 1999; Inoue et al., 2001). In addition, expression of the Arabidopsis thaliana MID1 COMPLEMENTING ACTIVITY 1 and 2 (MCA1,2) genes rescues a MID1-deficient yeast strain suggesting that they could function as stretch-activated calcium channels (Nakagawa et al., 2007; Yamanaka et al., 2010). Up to now no functional homologues for the yeast CWD sensors have been identified in plants. In yeast, the signals generated by the sensors are relayed to the response genes via different signaling cascades involving CALCINEURIN (MID1/CCH1); RHO1/MPK1 (WSC1, MID2), and YPD1 (SLN1) (Levin, 2005). Transcription factors mediating the response are SKN7, RLM1, and SWI4/6 (Levin, 2005). The response can involve activation of genes required for cell wall biosynthetic processes, remodeling of the cytoskeleton, and cell cycle progression. The available data suggest both organizational and functional similarities between the yeast and plant CWI maintenance mechanism while also highlighting how signals from mechanical and chemical sensors regulate jointly the CWD response.
Composition and Structure of the Plant Cell Wall
The plant cell wall is comparable to an exoskeleton surrounding the plant cell and providing both structural support and protection from biotic as well as abiotic stresses. It consists of cellulose microfibrils, pectin, hemicelluloses, proteins, and in certain cases lignin (Somerville et al., 2004). Plant cell walls are divided into primary (laid down during cell elongation/differentiation) and secondary (formed after cell morphogenesis is concluded) walls. In parallel, dependent on the presence of certain polysaccharides type I and type II cell walls are distinguished (Popper et al., 2011). Cellulose microfibrils are the main load bearing elements, which are cross-linked to hemicelluloses and (in vitro) to pectin (Dick-Perez et al., 2011). Hemicelluloses and pectin also form direct links creating a matrix in which the microfibrils are embedded like the steel mesh in a concrete wall. Pectic polysaccharides like homogalacturonan (HG) are connected by calcium bridges between dimethyl-esterified parts of the molecules or through borate ester linkages in the case of rhamnogalacturonan II (RG II). They represent important cell wall components during morphogenesis facilitating cell expansion and plant–pathogen interaction (Hahn et al., 1981; Bellincampi et al., 1996). In the latter situation they are targeted by pathogen-derived cell wall degrading enzymes (polygalacturonases), which generate oligogalacturonides (OGAs) from HG (Hahn et al., 1981; Kars et al., 2005). Biologically active OGAs consist of chains of 9–15 galacturonic acid (GalA) monomers and can function as signaling molecules (see below).
During secondary cell wall formation monolignols (precursors for lignin) are secreted into the cell wall space and randomly cross-linked (Vanholme et al., 2010). The cross-linking is dependent on the availability of reactive oxygen species (ROS) generated by laccases and peroxidases. This process reinforces the wall against pathogen infection, waterproofs it, and increases structural integrity (Tronchet et al., 2010; Vanholme et al., 2010). Reduction of cellulose biosynthesis during primary cell wall formation through genetic or chemical means leads to lignin production (Ellis et al., 2002; Hamann et al., 2009). This highlights the ability of the cell to adapt to changes in cell wall composition and provides evidence for the existence of a CWI maintenance mechanism.
The Plant CWI Maintenance Mechanism as a Component of the Biotic Stress Response
Plant cell walls are capable of adjusting their composition and structure in response to pathogen infection (Dong et al., 2008). A mutation in (CELLULOSE SYNTHASEA3, CESA3) a subunit of the cellulose synthase complex leads to ectopic production of lignin, i.e., replacement of a missing load bearing cell wall component by another one (Cano-Delgado et al., 2000). Follow up studies found that inhibition of cellulose biosynthesis during primary cell wall formation either through mutations like constitutive expression of VSP1 (cev1) and ectopic lignification1 (eli1) or inhibitors such as isoxaben results in transcriptional activation of stress response mechanisms; ectopic production of ethylene (ET), salicylic acid (SA), jasmonic acid (JA), callose, and ROS as well as changes in cell wall composition and structure (Ellis et al., 2002; Cano-Delgado et al., 2003; Manfield et al., 2004; Hamann et al., 2009). The experiments also showed that the response to CWD consists of early and late stages reminiscent of the response to pathogen infection (Denness et al., 2011). During the early stage, ROS- and calcium-based signaling cascades are required for initiating the response to CWD (Denness et al., 2011). Interestingly, 1-aminocyclopropane-1-carboxylic acid (ACC, an ET precursor) and not ET itself seems to be acting as signaling substance during the early response to isoxaben with inhibition of cell expansion being an active process and not simply an automatic consequence of cellulose biosynthesis inhibition (Tsang et al., 2011). During the late stage, responses to CWD-like lignin deposition are initiated and the extent of lignin formation is apparently modulated by a negative feedback loop formed by ROS and JA (Denness et al., 2011). A combination of genetic and phenotypic analysis has implicated the NADPH oxidases RESPIRATORY BURST OXIDASE HOMOLOGD and F (RBOHD, F), the serine/threonine kinase OXIDATIVE SIGNAL INDUCIBLE1 (OXI1), MCA1, the receptor-like kinase (RLK) THESEUS1 (THE1) as well as the JA biosynthesis genes ALLENE OXIDE SYNTHASE (AOS) and JASMONIC ACID RESISTANT1 (JAR1) in the signaling mechanism mediating the response to CWD in Arabidopsis seedlings. Interestingly, the cev1 mutation that affects cellulose biosynthesis during primary cell wall formation also causes enhanced resistance to infection by different powdery mildews (Erysiphe orontii, E. cichoracearum, and Oidium lycopersicum; Ellis and Turner, 2001).
A screen for mutants causing resistance to powdery mildew infection provides further evidence of the close relationship between plant cell walls and pathogen resistance (Vogel and Somerville, 2000). Three of the powdery mildew resistance (PMR) mutants that have been identified on the molecular level affect genes involved in cell wall biosynthetic processes. PMR4 encodes a callose synthase, PMR5 a gene of unknown function required for pectin production and PMR6 a pectate lyase (Vogel et al., 2002, 2004; Nishimura et al., 2003). pmr4 resistance seems to be mediated via hyper-activation of SA signaling, whereas pmr5 and 6 resistance phenotypes are independent of JA, SA, and ET signaling. Mutations in IRREGULARXYLEM1 (IRX1/CESA8), 3 (IRX3/CESA7), and 5 (IRX5/CESA4) impair cellulose biosynthesis during secondary cell wall formation and cause enhanced resistance to the soil borne bacterium Ralstonia solanacearum and the necrotrophic fungus Plectosphaerella cucumerina (Hernandez-Blanco et al., 2007). Mutations affecting cellulose biosynthesis during primary cell wall formation (cesa1, 3, 6) or other components of the secondary cell wall (pmr5, pmr6) did not cause enhanced resistance to the same pathogens. Genetic analysis showed that the enhanced resistance in cesa4, 7, 8 is independent of JA, SA, and ET-based signaling mechanisms. Results from expression profiling experiments and genetic analysis using different abscisic acid (ABA)mutants(ABA insensitive 1-1;2-1;ABA1-6) suggest that ABA is mediating developmental and pathogen resistance phenotypes caused by the irx mutants. However, it remains to be determined if the ABA involvement is direct or a secondary effect due to water stress caused by problems with xylem cell wall formation in the mutants. To summarize, these results suggest distinct resistance signaling cascades are induced by defects in primary and secondary cell wall formation as well as for different secondary cell wall components. They also highlight the direct impact of changes in cell wall composition/structure on the response to pathogen infection.
Signaling Mechanisms and Sensors Implicated in Plant CWD Perception
The mode of action of the plant cell wall maintenance mechanism is not well understood. Based on the knowledge from yeast, chemical and physical signals could act as indicators for the functional integrity of the plant cell wall either individually or jointly. By combining these different types of signal the plant cell would receive precise information regarding the state of its cell wall and the exact type of CWI impairment occurring. Physical signals could be generated by stretching of the plasma membrane due to a weakened cell wall that cannot resist the high turgor pressure levels within a plant cell or a plasma membrane that is displaced relatively to the cell wall. These events could be detected by stretch-activated or mechanosensitive channel proteins that lead to calcium influx into the cytoplasm, indicating CWD. Sensor candidates could be encoded by members of the mechanosensitive channels of small conductance (MscS)-like (MSL) gene family like MSL 9 and 10 affect mechano-perception in protoplasts derived from Arabidopsis root cells (Haswell et al., 2008). Another candidate of interest is the putative stretch-activated calcium channel MCA1 is required for CWD-induced lignin deposition. Interestingly, all isoxaben-induced CWD phenotypes can be suppressed by provision of osmotic support suggesting that changes in turgor pressure due to a weakened cell wall could result in signal generation via turgor pressure sensors (Hamann et al., 2009; Denness et al., 2011). While AHK1 and 4/CRE1 can function as osmosensors in yeast and have been implicated in abiotic stress responses, no clear evidence exists implicating them in CWD perception in plants (Urao et al., 1999; Inoue et al., 2001; Tran et al., 2007). In addition, AHK4/CRE1 has been shown to function as a cytokinin receptor(Inoue et al., 2001). Therefore, the question that needs to be resolved at this point is if turgor pressure is a passive element in the process (generating cell wall fragments due to a weakened cell wall) or an active component that is being monitored and provides input into the process.
The plant cell wall contains a large number of components that could generate chemical signals (ligands) indicative of CWD or general danger signals. The term damage associated molecular patterns (DAMPs) has been coined to describe such ligands and the number of possible DAMPs originating in plant cell walls is rather large (Zipfel, 2009). Here I will focus on the best-characterized group of signals, which are probably OGAs. They can be generated through degradation of HG by pathogen-derived enzymes (Kars et al., 2005; Ferrari et al., 2008). OGAs have been shown to induce gene expression changes, stomatal closure, production of ET, and ROS as well as cell wall reinforcement (Denoux et al., 2008; Ferrari et al., 2008). A hybrid kinase consisting of the extra cellular domain of WALL-ASSOCIATED KINASE1 (WAK1) and the intracellular domain of elongation factor Tu receptor (EFR) kinase can bind OGAs and activate defense responses (Brutus et al., 2010). WAK1 belongs to a family of five WAK genes encoding plasma membrane-localized Ser/Thr kinases that have been implicated in response to pathogen infection and regulation of cell elongation (Kohorn et al., 2011). The effects of the chimeric WAK1 kinase on pathogen resistance suggest that OGAs and WAKs represent an in vivo ligand–receptor pair. Results from the analysis of a dominant active WAK2 allele suggest the CWD signals perceived by WAKs could be relayed to downstream response genes through MAPKINASE6 (MPK6) (Kohorn et al., 2009, 2011). Interestingly, WAK2 has also been implicated in regulation of invertase activity and turgor pressure during cell elongation (Kohorn et al., 2006). However, there is currently no confirmation that WAKs are actively involved in CWI maintenance.
In Arabidopsis, more than 600 RLKs have been identified and a large number of them have been implicated in developmental and stress response processes (Shiu and Bleecker, 2001). I will focus here on several kinases that have been implicated in CWD perception and/or pathogen response. Most of the RLKs implicated in CWI maintenance [THE1, HERCULES1 (HERK1), FERONIA (FER)] belong to the Catharanthus roseus RLK1 (CrRLK1)-like protein family, which has 17 members in Arabidopsis. THE1 was isolated as a suppressor of the cellulose-deficient cesa6 procuste (pre) mutant, which exhibits a hypocotyl elongation defect (Hematy et al., 2007). Although thel suppresses the elongation defect, the cellulose deficiency is not reduced. Subsequently it has been shown that THE1 is required for cellulose biosynthesis inhibition-induced ROS production and lignification in the root elongation zone (Denness et al., 2011). THE1, HERK1, and FER have been implicated in brassinosteroid-induced cell elongation (Guo et al., 2009; Deslauriers and Larsen, 2010). Both FER and NORTIA/MILDEW RESISTANCE LOCUS O 7 (ML07; a seven-transmembrane domain protein involved in powdery mildew resistance) are required for successful fertilization and resistance to infection by Golovinomyces (syn. Erysiphe) orontii (Kessler et al., 2010). Interestingly, ROPGEF (guanine-exchange factors) proteins have been identified as targets of FER activity (Duan et al., 2010). ROPGEFs are required for the activation of Rho GTPases, which in turn activate NADPH oxidases like RBOHD/F. These results suggest the same molecular components could mediate cell-cell interaction during development and plant–pathogen interaction.
Heterotrimeric G-proteins (Gα, Gβ, Gγ) form a highly conserved signaling complex that has been implicated in signal transduction during development and stress responses in mammals, yeast, and plants (Digby et al., 2006; Temple and Jones, 2007). In Arabidopsis, five genes GPA1 (Gα), AGB1 (Gβ), AGG1, 2, 3 (Gγ1, 2, 3) encode the subunits of the complex (Thung et al., 2011). Recently, it has been reported that mutations in AGG1, 2, and AGB1 apparently cause enhanced susceptibility to infection with P. cucumerina (Delgado-Cerezo et al., 2011). A combination of metabolomic and microarray-based expression profiling studies of the mutants established that the pathogen phenotype is independent of SA, JA, ABA, and ET signaling cascades. Interestingly, a large number of cell wall biosynthetic/modifying genes are mis-regulated in agbl and aggl2 plants. Analysis of cell wall composition/structure in these plants found reduced xylose contents in the mutants compared to wildtype.
To summarize, the available evidence supports the notion that the plant cell wall is an integral component contributing to pathogen response mechanisms and illustrates the influence of cell wall defects on infection. Specific signaling cascades seem to mediate the response to particular cell wall defects, which in turn affect the response to necrotrophic or biotrophic pathogens. More importantly the data presented above allow correlation between certain types of cell wall defects, and not only signaling cascades but also resistance phenotypes. Mutations in CESA4, 7, 8, AGG1, 2, and AGB1 affect resistance to necrotrophs and are independent of phytohormone-based signaling cascades. pmr5, 6 plants exhibit resistance to biotrophs and also do not rely on phytohormone-based signaling cascades. pmr4 affects resistance to biotrophs and resistance depends on the integrity of the SA signaling cascade. cesa3 plants show enhanced JA, ET biosynthesis, and resistance to biotrophs.
The available data allow different explanations for the specific effects on pathogen resistance observed. The cell wall composition/structure changes could prevent pathogen colonization simply because the infection machinery of the pathogen is too specialized to breach the chemically modified cell wall. However, no functional pathogen response mechanisms should be required for this possibility. Another option is that the cell wall mutants cause defects similar to those occurring during infection by particular pathogens, i.e., simulate infection by necrotrophs or biotrophs. This would cause early/constant activation of the CWI maintenance/defense mechanism, which "primes" plant immunity thus making successful infection more difficult. The latter would explain both the specificity of the responses observed and dependence on particular signaling mechanisms. Therefore, studies focusing on the effects of particular cell wall defects on pathogen resistance and the mode of action of the CWI maintenance mechanism could facilitate research into biotic stress response. The reason being, that by removing the potentially multiple effects of the pathogen during infection, they reduce the complexity of the interaction and should therefore allow novel insights into the mechanisms responsible for detection of infection and/or physical damage.
Conflict of Interest Statement
The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
The author would like to thank John Mansfield and Issariya Chairam for critical reading of the manuscript and helpful comments. Research in the Hamann lab is supported by the Gatsby Foundation, BBSRC, and the FP7 framework program.
Bellincampi, D., Cardarelli, M., Zaghi, D., Serino, G., Salvi, G., Gatz, C., Cervone, F., Altamura, M. M., Costantino, P., and Lorenzo, G. D. (1996). Oligogalacturonides prevent rhizogenesis in rolB-transformed tobacco explants by inhibiting auxin-induced expression of the rolB gene. Plant Cell 8, 477–487.
Brutus, A., Sicilia, F., Macone, A., Cervone, F., and De Lorenzo, G. (2010). A domain swap approach reveals a role of the plant wall-associated kinase 1 (WAK1) as a receptor of oligogalacturonides. Proc. Natl. Acad. Sci. U.S.A. 107, 9452–9457.
Cano-Delgado, A., Penfield, S., Smith, C., Catley, M., and Bevan, M. (2003). Reduced cellulose synthesis invokes lignification and defense responses in Arabidopsis thaliana. Plant J. 34, 351–362.
Cano-Delgado, A. I., Metzlaff, K., and Bevan, M. W. (2000). The eli1 mutation reveals a link between cell expansion and secondary cell wall formation in Arabidopsis thaliana. Development 127, 3395–3405.
Delgado-Cerezo, M., Sanchez-Rodriguez, C., Escudero, V., Miedes, E., Fernandez, P. V., Jorda, L., Hernandez-Blanco, C., Sanchez-Vallet, A., Bednarek, P., Schulze-Lefert, P., Somerville, S., Estevez, J. M., Persson, S., and Molina, A. (2011). Arabidopsis heterotrimeric G-protein regulates cell wall defense and resistance to necrotrophic fungi. Mol. Plant 5, 98–114.
Denness, L., McKenna, J. F., Segonzac, C., Wormit, A., Madhou, P., Bennett, M., Mansfield, J., Zipfel, C., and Hamann, T. (2011). Cell wall damage-induced lignin biosynthesis is regulated by a ROS- and jasmonic acid dependent process in Arabidopsis thaliana. Plant Physiol. 156, 1364–1374.
Denoux, C., Galletti, R., Mammarella, N., Gopalan, S., Werck, D., De Lorenzo, G., Ferrari, S., Ausubel, F. M., and Dewdney, J. (2008). Activation of defense response pathways by OGs and Flg22 elicitors in Arabidopsis seedlings. Mol. Plant 1, 423–445.
Deslauriers, S. D., and Larsen, P. B. (2010). FERONIA is a key modulator of brassinosteroid and ethylene responsiveness in Arabidopsis hypocotyls. Mol. Plant 3, 626–640.
Dick-Perez, M., Zhang, Y., Hayes, J., Salazar, A., Zabotina, O. A., and Hong, M. (2011). Structure and interactions of plant cell-wall polysaccharides by two- and three-dimensional magic-angle-spinning solid-state NMR. Biochemistry 50, 989–1000. doi: 10.1021/bi101795q
Digby, G. J., Lober, R. M., Sethi, P. R., and Lambert, N. A. (2006). Some G protein heterotrimers physically dissociate in living cells. Proc. Natl. Acad. Sci. U.S.A. 103, 17789–17794.
Dong, X., Hong, Z., Chatterjee, J., Kim, S., and Verma, D. P. (2008). Expression of callose synthase genes and its connection with NPR1 signaling pathway during pathogen infection. Planta 229, 87–98.
Duan, Q., Kita, D., Li, C., Cheung A. Y., and Wu, H. M. (2010). FER-ONIA receptor-like kinase regulates RHO GTPase signaling of root hair development. Proc. Natl. Acad. Sci. U.S.A. 107, 17821–17826.
Ellis, C., Karafyllidis, I., Wasternack, C., and Turner, J. G. (2002). The Arabidopsis mutant cev1 links cell wall signaling to jasmonate and ethylene responses. Plant Cell 14, 1557–1566.
Ellis, C., and Turner, J. G. (2001). The Arabidopsis mutant cev1 has constitutively active jasmonate and ethylene signal pathways and enhanced resistance to pathogens. Plant Cell 13, 1025–1033.
Ferrari, S., Galletti, R., Pontiggia, D., Manfredini, C., Lionetti, V., Bellincampi, D., Cervone, F., and De Lorenzo, G. (2008). Transgenic expression of a fungal endopolygalacturonase increases plant resistance to pathogens and reduces auxin sensitivity. Plant Physiol. 146, 669–681.
Guo, H., Li, L., Ye, H., Yu, X., Algreen, A., and Yin, Y. (2009). Three related receptor-like kinases are required for optimal cell elongation in Arabidopsis thaliana. Proc. Natl. Acad. Sci. U.S.A. 106, 7648–7653.
Hahn, M. G., Darvill, A. G., and Albersheim, P. (1981). Host-pathogen interactions: XIX. The endogenous elicitor, a fragment of a plant cell wall polysaccharide that elicits phytoalexin accumulation in soybeans. Plant Physiol. 68, 1161–1169.
Hamann, T., Bennett, M., Mansfield, J., and Somerville, C. (2009). Identification of cell-wall stress as a hexose-dependent and osmosensitive regulator of plant responses. Plant J. 57, 1015–1026.
Hamann, T., and Denness, L. (2011). Cell wall integrity maintenance in plants: lessons to be learned from yeast? Plant Signal. Behav. 6, 1–5.
Haswell, E. S., Peyronnet, R., Barbier-Brygoo, H., Meyerowitz, E. M., and Frachisse, J. M. (2008). Two MscS homologs provide mechanosensitive channel activities in the Arabidopsis root. Curr. Biol. 18, 730–734.
Hematy, K., Cherk, C., and Somerville, S. (2009). Host–pathogen warfare at the plant cell wall. Curr. Opin. Plant Biol. 12, 406–413.
Hematy, K., Sado, P. E., Van Tuinen, A., Rochange, S., Desnos, T., Balzergue, S., Pelletier, S., Renou, J. P., and Hofte, H. (2007). A receptorlike kinase mediates the response of Arabidopsis cells to the inhibition of cellulose synthesis. Curr. Biol. 17, 922–931.
Hernandez-Blanco, C., Feng, D. X., Hu, J., Sanchez-Vallet, A., Deslandes, L., Llorente, F., Berrocal-Lobo, M., Keller, H., Barlet, X., Sanchez-Rodriguez, C., Anderson, L. K., Somerville, S., Marco, Y., and Molina, A. (2007). Impairment of cellulose synthases required for Arabidopsis secondary cell wall formation enhances disease resistance. Plant Cell 19, 890–903.
Humphrey, T. V., Bonetta, D. T., and Goring, D. R. (2007). Sentinels at the wall: cell wall receptors and sensors. New Phytol. 176, 7–21.
Inoue, T., Higuchi, M., Hashimoto, Y., Seki, M., Kobayashi, M., Kato, T., Tabata, S., Shinozaki, K., and Kakimoto, T. (2001). Identification of CRE1 as a cytokinin receptor from Arabidopsis. Nature 409, 1060–1063.
Kars, I., Krooshof, G. H., Wagemakers, L., Joosten, R., Benen, J. A., and van Kan, J. A. (2005). Necrotizing activity of five Botrytis cinerea endopolygalacturonases produced in Pichia pastoris. Plant J. 43, 213–225.
Kessler, S. A., Shimosato-Asano, H., Keinath, N. F., Wuest, S. E., Ingram, G., Panstruga, R., and Grossniklaus, U. (2010). Conserved molecular components for pollen tube reception and fungal invasion. Science 330, 968–971.
Kohorn, B. D., Johansen, S., Shishido, A., Todorova, T., Martinez, R., Defeo, E., and Obregon, P. (2009). Pectin activation of MAP kinase and gene expression is WAK2 dependent. Plant J. 60, 974–982.
Kohorn, B. D., Kobayashi, M., Johansen, S., Riese, J., Huang, L. F., Koch, K., Fu, S., Dotson, A., and Byers, N. (2006). An Arabidopsis cell wall-associated kinase required for invertase activity and cell growth. Plant J. 46, 307–316.
Kohorn, B. D., Kohorn, S. L., Todorova, T., Baptiste, G., Stansky, K., and McCullough, M. (2011). A dominant allele of Arabidopsis pectin-binding wall-associated kinase induces a stress response suppressed by MPK6 but not MPK3 mutations. Mol. Plant. doi: 10.1093/mp/ssr096
Levin, D. E. (2005). Cell wall integrity signaling in Saccharomyces cerevisiae. Microbiol. Mol. Biol. Rev. 69, 262–291.
Manfield, I. W., Orfila, C., McCartney, L., Harholt, J., Bernal, A. J., Scheller, H. V., Gilmartin, P. M., Mikkelsen, J. D., Paul Knox, J., and Willats, W. G. (2004). Novel cell wall architecture of isoxaben-habituated Arabidopsis suspension-cultured cells: global transcript profiling and cellular analysis. Plant J. 40, 260–275.
Nakagawa, Y., Katagiri, T., Shinozaki, K., Qi, Z., Tatsumi, H., Furuichi, T., Kishigami, A., Sokabe, M., Kojima, I., Sato, S., Kato, T., Tabata, S., Iida, K., Terashima, A., Nakano, M., Ikeda, M.,Yamanaka, T., and Iida, H. (2007). Arabidopsis plasma membrane protein crucial for Ca2+ influx and touch sensing in roots. Proc. Natl. Acad. Sci. U.S.A. 104, 3639–3644.
Nishimura, M. T., Stein, M., Hou, B. H., Vogel, J. P., Edwards, H., and Somerville, S. C. (2003). Loss of a callose synthase results in salicylic acid-dependent disease resistance. Science 301, 969–972.
Popper, Z. A., Michel, G., Herve, C., Domozych, D. S., Willats, W. G., Tuohy, M. G., Kloareg, B., and Stengel, D. B. (2011). Evolution and diversity of plant cell walls: from algae to flowering plants. Ann. Rev. Plant Biol. 62, 567–590.
Ringli, C. (2010). Monitoring the outside: cell wall-sensing mechanisms. Plant Physiol. 153, 1445–1452.
Seifert, G. J., and Blaukopf, C. (2010). Irritable walls: the plant extra cellular matrix and signaling. Plant Physiol. 153, 467–478.
Shiu, S. H., and Bleecker, A. B. (2001). Receptor-like kinases from Arabidopsis form a monophyletic gene family related to animal receptor kinases. Proc. Natl. Acad. Sci. U.S.A. 98, 10763–10768.
Somerville, C., Bauer, S., Brininstool, G., Facette, M., Hamann, T., Milne, J., Osborne, E., Paredez, A., Persson, S., Raab, T., Vorwerk, S., and Youngs, H. (2004). Toward a systems approach to understanding plant cell walls. Science 306, 2206–2211.
Temple, B. R., and Jones, A. M. (2007). The plant heterotrimeric G-protein complex. Ann. Rev. Plant Biol. 58, 249–266.
Thung, L., Trusov, Y., Chakravorty, D., and Botella, J. R. (2011). Gγ1 + Gγ2 + Gγ3 = Gβ: the search for heterotrimeric G-protein gamma subunits in Arabidopsis is over. J. Plant Physiol. 169, 542–545.
Tran, L. S., Urao, T., Qin, F., Maruyama, K., Kakimoto, T., Shinozaki, K., and Yamaguchi-Shinozaki, K.(2007). Functional analysis of AHK1/ATHK1 and cytokinin receptor histidine kinases in response to abscisic acid, drought, and salt stress in Arabidopsis. Proc. Natl. Acad. Sci. U.S.A. 104, 20623–20628.
Tronchet, M., Balague, C., Kroj, T., Jouanin, L., and Roby,D.(2010). Cinnamyl alcohol dehydrogenases-C and D, key enzymes in lignin biosynthesis, play an essential role in disease resistance in Arabidopsis. Mol. Plant Pathol. 11, 83–92.
Tsang, D. L., Edmond, C., Harrington, J. L., and Nuhse, T. S. (2011). Cell wall integrity controls root elongation via a general 1-aminocyclopropane-1-carboxylic acid-dependent, ethylene-independent pathway. Plant Physiol. 156, 596–604.
Urao, T., Yakubov, B., Satoh, R., Yamaguchi-Shinozaki, K., Seki, M., Hirayama, T., and Shinozaki, K. (1999). A transmembrane hybrid-type histidine kinase in Arabidopsis functions as an osmosensor. Plant Cell 11, 1743–1754.
Vanholme, R., Demedts, B., Morreel, K., Ralph, J., and Boerjan, W. (2010). Lignin biosynthesis and structure. Plant Physiol. 153, 895–905.
Vogel, J., and Somerville, S. (2000). Isolation and characterization of powdery mildew-resistant Arabidopsis mutants. Proc. Natl. Acad. Sci. U.S.A. 97, 1897–1902.
Vogel, J. P., Raab, T. K., Schiff, C., and Somerville, S. C. (2002). PMR6, a pectate lyase-like gene required for powdery mildew susceptibility in Arabidopsis. Plant Cell 14, 2095–2106.
Vogel, J. P., Raab, T. K., Somerville, C. R., and Somerville, S. C. (2004). MutationsinPMR5 result in powdery mildew resistance and altered cell wall composition. Plant J. 40, 968–978.
Wolf, S., Hematy, K., and Hofte, H. (2011). Growth control and cell wall signaling in plants. Ann. Rev. Plant Biol. doi: 10.1146/annurev-arplant-042811-105449
Yamanaka, T., Nakagawa, Y., Mori, K., Nakano, M., Imamura, T., Kataoka, H., Terashima, A., Iida, K., Kojima, I., Katagiri, T., Shinozaki, K., and Iida, H. (2010). MCA1 and MCA2 that mediate Ca2+ uptake have distinct and overlapping roles in Arabidopsis. Plant Physiol. 152, 1284–1296.
Zipfel, C. (2009). Early molecular events in PAMP-triggered immunity. Curr. Opin. Plant Biol. 12, 414–420.
Keywords: plant cell wall integrity, biotic stress, plant pathogen
Citation: Hamann T (2012) Plant cell wall integrity maintenance as an essential component of biotic stress response mechanisms. Front. Plant Sci. 3:77. doi: 10.3389/fpls.2012.00077
Received: 27 January 2012; Accepted: 05 April 2012;
Published online: 23 April 2012.
Edited by:Jose Manuel Estevez, University of Buenos Aires and CONICET, Argentina
Reviewed by:Kian Hématy, Institut National de la Recherche Agronomique, France
Staffan Persson, Max-Planck-Gesellschaft, Germany
Copyright: © 2012 Hamann. This is an open-access article distributed under the terms of the Creative Commons Attribution Non Commercial License, which permits non-commercial use, distribution, and reproduction in other forums, provided the original authors and source are credited.
*Correspondence: Thorsten Hamann, Division of Cell and Molecular Biology, Department of Life Sciences, Imperial College London, Sir Alexander Fleming Building (608), South Kensington Campus, London SW7 2AZ, UK. e-mail: firstname.lastname@example.com


text:
The following is the first of a series of interviews launched for Fetish Portraiture photography week in Project Educate 2008.
5yearcheddar is a staple of the deviantART fetish (and photography in general) community, having been on the site for 5 years now. His work reflects a level of skill to which many photographers aspire and not all obtain. I've been a fan for ages, so I was really excited that he agreed to subject himself to my battery of questions!
Basics and Background
Name and location?
Chris Young. I currently reside in NE Wisconsin, USA.
How long have you been practicing photography?
For as long as I can remember. The first camera I remember having was a kodak instamatic with a zoom lens. Anyone remember those? I didn't really start shooting seriously until the late 80's when I "inherited" a 35mm SLR and had to learn how to use it. The rest as they say is history.
Do you have any formal trai

subset: commoncrawl


text:
- How are reviewers describing this item?
great, good, cozy, little and perfect.
- Our engine has detected that Amazon has altered and removed reviews from this listing. We approximate total reviews removed to be 360.
- Our engine has profiled the reviewer patterns and has determined that there may be deception involved.
- Our engine has determined that the review content quality is low.
- Our engine has analyzed and discovered that 63.9% of the reviews are reliable.
- This product had a total of 682 reviews as of our last analysis date on Feb 19 2019.
Reviews Summary
Bought for my daughter's apple pen
This these items are a must have for any apple pencil owner
00?it costs cost me to replace my pencil hag i purchased these inexpensive protective items
Now i feel at ease knowing that i will not lose my apple pencil parts
Review Count History
Price History


text:
By James C. Whorton
this can be the 1st complete background of different drugs in the United States, analyzing the most important platforms that experience emerged from 1800 to the current. Writing with wit and with equity to either side, Whorton bargains a desirable examine replacement future health platforms comparable to homeopathy, water remedies, Mesmerism, Christian technological know-how, osteopathy, chiropractic, naturopathy, and acupuncture. He highlights the start and development of every approach (including eu roots the place applicable) and vividly describes either the theories and the cures built inside of each one method, together with such doubtful practices as hour-long walks barefoot in snow or Samuel Thompson's "puking and steaming" routine. particularly, Whorton illuminates the philosophy of "natural therapeutic" that has been espoused via replacement practitioners all through heritage and the precise interpretations of "nature therapy" built through the various platforms. notwithstanding he does not hesitate to show the flaws of those structures, he additionally indicates that a few "cult medicinal drugs" have ultimately received popularity from practitioners of mainstream medicine.
all through, Whorton writes with a gentle contact and costs from modern humorists akin to Mark Twain. His booklet is a fascinating and authoritative heritage that highlights the process substitute drugs within the united states, supplying priceless heritage to the wide variety of remedies on hand today.
Read Online or Download Nature Cures: The History of Alternative Medicine in America PDF
Best alternative medicine books
Get The Miraculous Achievements of Bodywork: How Touch Can PDF
Why do a little humans miraculously flip round after struggling with existence threatening health problems, emotional trauma, and non secular melancholy? Dr. Ronan Kisch has documented twenty-seven bodywork/massage therapists whose clients—despite grave clinical prognoses or existence trauma—had strange, if no longer mind-blowing recoveries after having bodywork with those practitioners.
Read e-book online Schüßler-Salze: Die fantastischen 12 (German Edition) PDF
Vitaler Körper - entspannte Seele Die Schüßler-Therapeutin Sabine Wacker staunt immer wieder über die kleinen Wunder, die sie mit der "Schüßlerei" erlebt und nennt die 12 biochemischen Mineralstoffe nach Dr. Schüßler deshalb die fantastischen 12. Dabei gibt es die Schüßler-Salze schon seit über a hundred thirty five Jahren.
Meet Jamie Now: A Life Free of Autism - download pdf or read online
Meet Jamie now could be a daring, intimate examine a girl as she fights for the loves of her existence. After one in all her sons, Jamie, is clinically determined with autism, all that she loves is threatened by means of her relentless pursuit of a medication. Krista is a traditional storyteller who attracts the reader into her existence and the lives of her family members.
Download e-book for kindle: 99+ International Cancer Preventers, Cancer Fighters, Cancer by Joseph A. Laydon Jr.
Your physique has the innate skill to heal itself. And it does heal itself time and again an afternoon simply because your immune procedure is operating each break up moment 24-hours an afternoon fighting-off every kind of undesirable men in and out your physique. besides the fact that, within the usa on my own, greater than 500,000 american citizens die from melanoma every year.
- The Lawrence Metamorphosis Mediterranean Heart Attack and Stroke Prevention Weight Loss Diet Program (Lawrence Metamorphosis Dietary Programs Book 2)
- Health For US All
- Generational Emotional Mapping: Reprogramming the Subconscious with Essential Oils
- Medical Intuition: Your Awakening to Wholeness
- The Reprogramming: Detox Your Mind's Negativity and Create the Life you Want
Extra info for Nature Cures: The History of Alternative Medicine in America
Example text
Nature Cures: The History of Alternative Medicine in America by James C. Whorton
by Richard


text:
How Neurocore Is Combating Depression & Mental Illness
Neurocore may not be the first name that springs to mind when it comes to battling depression and a number of other mental health issues, but over the last few years, the Michigan based company has become an innovative leader in the field. Since its foundation in 2004, the company has set up a significant number of Neurocore Brain Performance Centers across the country. Visit Patch.com to know more about Neurocore.
Since opening, these centers have specialized in data-driven, brain-based diagnostics and treatments for depression and it's symptoms; their techniques not only diagnose and treat the major symptoms but have shown some amazing accuracy when it comes to the minor symptoms. These tend to be almost completely invisible to people who aren't affected by mental illness, which has made the companies techniques pretty groundbreaking. Because of these techniques the company has become somewhat of a leader in the field of applied neurofeedback. This field not only helps with symptoms of depression but also improves concentration, helps one to sleep better, and has been shown to help people manage stress.
Check: https://www.crunchbase.com/organization/neurocore
Neurocore clinicians use advanced qEEG technology to measure and evaluate a patients brain electrical activity to identify the cause of their depression symptoms. Combined with some other diagnostic tools, this gives patients and medical practitioners unprecedented access to a complete picture of a patients brain. With these results, Neurocore can then create a more personal Neurotherapy treatment for their patient to combat their depression. Their treatments are said to be groundbreaking due to the accuracy of their qEEG technology compared to more traditional behavioral checklist diagnostic tools.
Because of the accuracy of the diagnoses and treatments, it looks likely that many mental health professionals will use these tools to supplement the more traditional tools used to diagnose and treat depression. Because of that, it looks like Neurocore is leading the march to combat mental health issues. Follow Neurocore on Twitter.


text:
Ironically, technology can make the utilities' problem worse, not better—at least in the short term. In the past, grids were developed because it was cheaper to generate large quantities of power and distribute it over wide distances, rather then generate smaller quantities closer to the place of use. It is for this reason that electricity is seen as a business that benefits from "economies of scale".
ElectricityPlans lets you easily compare electricity plans by displaying all-inclusive rates at all advertised usage levels in a simple easy-to-read format. You can easily estimate your actual all-in electric bill at any given usage level using our Plan Details and Pricing section for each plan. All energy charges, delivery fees, bill credits, and other fees for each plan are shown so you can accurately estimate your monthly electric bill. By showing all rates and fees, you'll avoid the electric bill sticker shock and so-called "teaser rates" commonly used by electric suppliers to achieve better search results on sites such as powertochoose.org.
Because prices from energy companies in Maryland rise and fall depending on the wholesale price of energy, residents may want a more stable rate for a longer period. A fixed-rate energy plan offers you just that, with a consistent rate for the duration of your agreement at current energy prices – regardless of fluctuations in the energy market. This could mean protection from rate spikes and savings for electric utility customers.
In summary, fixed-rate plans provide a level of certainty and stability in your energy charge since the price will not fluctuate over the life of your contract. If prices suddenly spike, you are protected because your rate is locked in. The flip side is that if rates drop over the life of your contract, you'll be stuck paying the higher rate. You can incur steep cancellation fees if you change electricity plans or providers before the end of your contract term.
Texas electricity deregulation has given millions of Houston residents and businesses the power to choose the cheapest electricity rate. According to ERCOT, over 92% of Texas homes and businesses who live in deregulated areas have switched electric companies since deregulation began in 2002. Even though electric choice in Texas has been hugely successful for energy savings, customers are still confused by the options, terminology, and overall process of switching electric providers.
Are you a journalist or researcher writing about this topic who needs to know more about historical rates? Send us details about what you need and we'll get back to you with an answer and a relevant quote from one of our rate experts. You should also check out the Choose Energy Data Center for more statistics and analysis centering on energy in the U.S.
You can sort, filter, and shop by pricing at YOUR specific usage level, which lets you shop and compare electricity plans based on the rates you'll actually experience on your bill, inclusive of hidden fees and taxes. This ensures you're not misled by the cheaper rates often advertised by electric providers…those "teaser rates" associated with higher usage levels that many households never enjoy because their usage level never reaches that pricing tier.
Utility companies are responsible for transmission and delivery of electricity even in energy deregulated parts of Texas and should be contacted in the event of a power outage. Your retail energy supplier may provide you competitive electric rates or exceptional customer service, but they cannot repair power lines or restore your service. In the case of an emergency, contact:
There are over 60 different energy suppliers competing for your business on any given day in Texas. Many of these electric companies have websites that are confusing and nearly impossible to navigate, their rates and fees hidden by dense industry jargon and misleading advertising. Who has the spare the time to sort through the choices spread out over all these different sites and companies?


text:
Futuristic Mid Century Modern Bedroom Furniture
For those who prefer futuristic, minimalist furniture for their bedroom, then mid century modern bedroom furniture can be the best choice. Mid-century modern furniture is known for its retro-futuristic elements
Mid Century Furniture Design
The Mid-century furniture design gave us clean lines, gentle organic curves, a love for different materials and designers that even today are perennial favorites. Mid-century furniture design is a term
Elegant Modern Furniture Design
Elegant modern furniture is sophisticated and practical modern furniture. Elegant modern furniture reflects a simplistic design style which emphasizes line and the use of both the furniture and the space
Modern Bedroom Furniture Styles
Everywhere you look you find things are being updated. The best way to start modernizing in your life is to have a modern bedroom. In modern bedroom, modern bedroom furniture
Modern Home Office Furniture
for modern home office furniture is a mid-shade wood, such as cherry, maple or oak, and a light colored wood floor. Add to that white or pale grey woodwork, mid-shade
Bamboo Furniture
thing, and not imitation bamboo furniture. You can get pretty much any piece of furniture you might need made out of bamboo. This furniture doesn't have to look like it
The Perfect Furniture For Small Spaces
of furniture singularly and assemble them on your own. Another option is to ask your carpenter or furniture maker to take measurements of the space and then make the furniture.
Cheap Living Room Furniture
furniture, it will usually be secondhand furniture. It will be the furniture one person is selling to buy new living room furniture. So logically, the best place to look for
Modern Kitchen Design
Modern kitchen design have become very popular in the present times. Modern kitchen design have a unique style and clean lines, which make people, love the modern kitchen design. If
Stylish Modern White Dining Chairs
and designs. Modern white dining chairs goes beautifully with other furniture colors and in fact makes other furniture colors look more attractive. The best part is modern white dining chairs
Remarkable Asian Style Furniture
of Asian style furniture is Japanese furniture. Japanese furniture features hard woods and clean lines. Until the 19th century, the Japanese didn't use chairs, so much of the furniture is
Alluring Victorian Style Furniture
Victorian style furniture refers to 19th-century furniture from the Victorian period. Victorian style furniture typically has intricate carving with curving lines and natural images such as leafy patterns, floral patterns,
Ahigo.net is not a retail shop, nor do we sell anything from this website. Any content, trademark's, or other material that might be found on this website that is not this site property remains the copyright of its respective owner's.


text:
The perfect camp for both new and experienced student performers, they will build skills in singing, dancing, and storytelling using their favorite current songs. The week ends with a showcase performance on the last day for families and friends. This camp is recommended for ages 8 - 13.
March 18 - 22, 2018
10am - 2pm, Monday - Friday
Tuition: $100
Showcase Performance on Friday, March 22th at 2:00pm.
Check out our Summer Theatre Camps for all ages
Sign-ups begin March 26th
More info here:
Call 772-231-6990 with any questions.


text:
Forget winter—handsomeness is coming!
Games of Thrones' Kit Harington has been tapped to front Jimmy Choo shoes' latest campaign, and boy, is it a glorious one.
The British actor, 28, who's best known for playing the sensitive bastard child Jon Snow in the HBO series, clearly channeled his tortured character in the ad photos and video—look at that stunning resting sad face! Only such a heart-melting expression could make the required sports car-climbing look like such a noble endeavor.
And yes, alright, there's also some fashion on display. One shot has the Night's Watchman rocking a blue suit and $800 high-shine metallic sneaks (while lounging on a windshield, natch). The other has the How to Train Your Dragon 2 star behind the wheel, still suited up but this time in printed driving loafers. Very practical in Westeros, they'd be.
The only thing missing: Harington's luscious, trademarked curls, snipped and slicked back to create a sleek, parted style. Hello, more face!
Do you love Harington's Jimmy Choo ads as much as we do? Tweet with @UsWeekly using the hashtag #stylebyUs!
Sign up now for the Us Weekly newsletter to get breaking celebrity news, hot pics and more delivered straight to your inbox!
Want stories like these delivered straight to your phone? Download the Us Weekly iPhone app now!


text:
Friday, 6 July 2018
Messy play and Skipper
12 comments:
- da tabbies o trout towne said...
...skipper...we noez how much ewe lovez de out o doorz but when itz vizshuz hot like that everee one wantz ta stay in...peepulz, wild life....ya noe.....sorree ya had ta go two de eeeeeeeeeeeevil place N heerz hopin ya keep on stayin better...stay inn side fora bit....tell mum ta put sum garfield on de telly for ya ~~~~~~ st francis' blessingz ♥♥♥
- 6 July 2018 at 19:44
Hi Anesha love what you did with the colours ,they look fantastic,and i am glad Skipper is feeling better xx
- 7 July 2018 at 02:22
- aussie aNNie said...
Beautiful backgrounds, well done Skipper.,.,hugs.
- 7 July 2018 at 03:03
- Dotty Jo x said...
Stunning backgrounds - how bright and cheery! Hope Skipper is doing ok. The weather is so hard on our pets, isn't it. Poor Custard is struggling with the heat and his diabetes. Hugs, Jo x
- 7 July 2018 at 11:00
- Paper Creations by Shirley said...
I see beautiful backgrounds from your play and good to hear that Skipper is doing well. It was VERY hot here as well. Take care, Shirleyx
- 7 July 2018 at 13:22
- Margarets designer cards said...
Love your colours and your play with cards.
Glad she recovering, hope it's nothing serious
- 7 July 2018 at 13:36
- cuilliesocks said...
Hi Anesha, Skipper is gorgeous, I hope she gets better, just too hot for her I guess.
Your backgrounds are fabulous, smashing colours, Kate x
- 7 July 2018 at 15:20
- Hazel (Didos) said...
Oh these backgrounds are super, love the bright colours. I hope SKipper is feeling a bit better today. Its so scary when they are old and get poorly, Sending her furry purrs and head butts from Felix and Belle, Love H x
- 7 July 2018 at 18:38
- Tamago said...
Those are very colorful and pretty.
So glad Skipper is doing well after vet visit. It's been so hot here as well. Stay cool and have a lovely weekend xo
- 7 July 2018 at 21:10
- Marina said...
Fabulous colour and pattern on your backgrounds. Hope Skipper is okay.
- 8 July 2018 at 16:57
- Andrea, said...
Great background and fab colours and designs. I hope Skipper is feeling better x
- 11 July 2018 at 08:53
- Maggie from Stillness at Cherith said...
Heat can be hard on our fur kids. I'm glad you are watching out for her. Glad to hear that she is feeling better.
Sending more purrs your way,sweet girl!
- 20 July 2018 at 18:29


text:
Forex Course
Currency Trading
Forex SystemPhone number is invalid FX Platform Category: Forex Backtesting ...or find similar posts on: Backtesting Results, Trending Pin Bar Backtesting © BKForex LLC 2015. All Rights Reserved Lecture3.3 Trade the Financial Markets Important Legal Notice: The information and opinions in this report are for general information use only and are not intended as an offer or solicitation with respect to the purchase of sale of any currency. All opinions and information contained in this report are subject to change without notice. This report has been prepared without regard to the specific investment objectives, financial situation and needs of any particular recipient. While the information contained herein was obtained from sources believed to be reliable, author does not guarantee its accuracy or completeness, nor does author assume any liability for any direct, indirect or consequential loss that may result from the reliance by any person upon any such information or opinions. Join the conversation WebMoney Search SpringerLink Lesson transcripts We have assembled an extensive list of the 20 best online Forex Courses for you to start your education in the exciting currency market. In addition, TD Ameritrade has mobile trading technology, allowing you to not only monitor and manage your forex position, but trade currencies right from your smartphone, mobile device, or iPad. forex trading station | forex trading system definition forex trading station | forex trading system development forex ab | forex trader là gì
Legal | Sitemap


text:
Wanting Nsa
Age 30 to 45. You have the great red hair, I've ever seen, and are beautiful. Let's do this tonight.
|Relationship Status:
|I Wants For A Man
|Killeen, TX
|Not important
|Relation Type:
|Adult Personals Ready I Want Free Sex
Views: 1640
Seeking fem sub I am looking for that one special lady who is submissive and bisexual and would like to meet a fantastic couple. We are looking for someone Wives want nsa Lorimor wants to explore her bisexuality and enjoys the best of both worlds.
Live sex dating Housewives want casual sex MD Leonardtown
Nas will be our sub and we all will role play. The male is Attractive very fit well hung and a super nice guy. The female is very fem classy and sub. You two will serve Beautiful housewives wants nsa Rockhampton and I will instruct you what I want you to Wives want nsa Lorimor. You will really enjoy this.
So if you have been looking explore Wives want nsa Lorimor submissive side we would love to talk with you. We are in the St. Charles area so someone close would be nice. Please no one liners or they will be deleted right away. Tell us about yourself and a pic would be nice too. We will get right back with you. I know she is out there.
Wives want nsa Lorimor I Am Looking Adult Dating
I am a professional woman with a great career that I enjoy. I don't have any kids, although I love them. I plan to have one or Wivees possibly through adoption.
I am considered a soft Lorinor in the lesbian world. I am very Wives want nsa Lorimor in the fact that I am woman but I do dress in men clothing. I am very much a professional with it. I am a god fearing and enjoy church every Sunday.
I have my own car as well as my own home. I am an lbs, nice build. I work out to keep it all together.
Iowa swingers contacts - free sex and dogging in Iowa, USA
I am looking to meet an intelligent femme woman between the ages of Please respond with a pic and Wives want nsa Lorimor will return the favor. Looking forward to hearing from you soon. I'll come to Wives want nsa Lorimor.
I am open to any age legal or Loorimor. Married and younger women move to the front of the line. You must be openminded and willing to please me wabt hesitation. If this is something you may be interested in reply and we'll see how much fun we can have.Las Palmas Sex Girls
I'm easy going I just know what I like and that is a woman who will do College girls fucking in wilmington I wish and then tell me about Wiges. Tomorrow is Memorial Day. Guarulhos sexy moms femme lesbians Things about myself Smart at times Adventurous A full-time college student I Wives want nsa Lorimor employed Movie buff! My height is 5'3 and I'm Wives want nsa Lorimor.
Llrimor Send a pic and a bit about yourself! Late 20's strung-out living on the street in San.
Mid 30's Following the telecom boom. Late 30's Partnered with a mid 30's drop-out whose clock is ticking hard. Mid 40's Stabbed in the back by a late 30's Wives want nsa Lorimor I whom I have wan all I have in this world.
Late 40's single father raising that have not seen their for almost a year.
I Ready Sexy Chat Wives want nsa Lorimor
We all have our journeys to Older married ladies only 45 to 60, I you on the other side.
These things seperate us from the lower lifeforms. Also it helps to Wives want nsa Lorimor out those carrying std's and Wives want nsa Lorimor carrying about those they pass it on to. To the guy who say's I'm hiding from family, wife and. A lot you know. I'm single, no and my family and friends know I'm bi. I enjoy women as well. There is a big difference between bi, and straight. The human heart isn't confined to just one gender. There is a test for women but not for men.
Often if it is HPV they tell you to wait to if it gets worse or goes away on it's own. Some people actually clear the virus on their own.Adult Friend Finder Lethbridge
More than 30 types are spread through sexual contact. Some types Wives want nsa Lorimor HPV cause genital warts single or multiple wan that appear in the genital areas of men and women, including the vagina, cervix, vulva the area outside of the vaginapenis, and rectum.
On the other hand, people infected with HPV have Wives want nsa Lorimor symptoms. Here's a good link to some more info; I was totally ignorant of HPV in men, how men actually have and spread it without knowing it and how easy it is to get it.Big Bear Lake Phone Chat
I also asked the Doctor about it as I was unaware of it. It just be a hemorrhoid but you Local guy seeking lady have shown it to the Doctor, then you would know. I'm not being a. I'm very aware of my health and never take it for granted. I was with a guy for so that was all wrong Wives want nsa Lorimor me. I still like guys but right now I'm taking Wives want nsa Lorimor break from them becuase I'm sick of them.
Also when I was in school, I always liked guys so this is more like something I only realized when I was in my 20s. All I can tell you is keep an open mind. Some of my friends are bi so I've been talking to them and asking them for advice. Good luck Hico free horny women in Chullu Vtoroye woman-hating bitter dudes. Secondly, your idea of getting a job and leaving is very sensible. You might have to consider moving to a bigger city or better state. Start now to build up your new social life where he is Wives want nsa Lorimor welcome or around.
Think in terms of present and future benefits to yourself as a single.
I assume you don't have. Keep it rational and polite. Good luck on that. Mature massage Trapper Creek. Virgin seeking milf or cougarwoman MFW. LormiorAdult Dating MA Cambridge 2139
Ready and willing Hello I'm looking to have some no strings attach fun with a lady. Please be DDD free and discreet.
Nas be over the age of Can't wait to hear from you. Send a with your response. Younger looking for older or someone Wives want nsa Lorimor If you are sitting there and you are older and think we might not have a lot in common try me you will Yerkes KY adult swingers surprised.
First off I am not into drama nor will I play any games or bull shit.
I am looking for classy woman who looks good takes care of herself likes to have Wives want nsa Lorimor and looking for a man not a boy. I want someone who can go out with me have fun and enjoy each others company and someone that wants to take the time to get to know me. Wives want nsa Lorimor am 6'3 lb br hair br eyes and I have a beautiful smile and I take care of myself I am at the gym all the time and I play too many sports.
I dont mind reading a book dancing cooking or anything like that because I like that stuff too. I like to do so many things Free live adult chat lines 89005 this world it isnt funny I am outside all the time so pretty much anything outside is fun to me.
I am very mature I dont drink much I will have a beer here and there and I dont smoke I dont like to party. I watn I havent giving you much incite on me but what would the Wives want nsa Lorimor of that be by me telling you everything I like to do?
I am not posting a pic on here due to work and school. Lorlmor am Wives want nsa Lorimor today is the 15th Ass slapping in West Mississippi your favorite restaurant in the title so I know your real Women who likes to fuck Kittitas Washington la Wives want nsa Lorimor reason and your passion are the rudder and the sails of your seafaring soul.
If either your sails or your rudder be broken, you can but toss and drift, or be held at a standstill in mid-seas. For reason, ruling alone, is a force confining; and passion, unattended, is a flame that burns to its own destruction. Therefore let your soul exalt your reason to the height of passion, that it sing; And let it nsw your passion with reason, that your passion live through its own daily resurrection, and like the phoenix, rise above its own ashes.
Black man looking Wives want nsa Lorimor pleasure you. She probably is working, taking care of the, cooking, doing grocery shopping, and other house chores,no time to raise another kid, You. How come you have so much time to waste your time doing unimpressive things like that?


text:
Lead Tax Advisor (Direct Tax)4 months ago - Sydney - 63 views
Job Details
Lead Tax Advisor (Direct Tax)
Req ID 95229BR Job category Tax Countries (State/Region) Victoria Location Australia - Victoria - Melbourne Apply now
Role synopsis
We have a unique 12-month opportunity to join our tax team as a senior member advising our downstream businesses. This role supports our Downstream businesses across Australia and New Zealand with optimising the group's tax position and risk.
BP is dynamic and growing, presenting you with a diversity of work and an opportunity to stretch your capability and allowing you to apply your skills in a deep and rewarding environment. This role predominantly advising on income taxes, but also dealing with cross boarder transfer pricing, M&A activity, Shipping, Permanent establishments and capital gains tax. This role is critical in providing guidance to the business in accordance with BP's approach to tax ensuring compliance with legislation.
This position reports to the Head of Tax for Australia & New Zealand.
Key accountabilities
- Strategic partner to the business
- Contribute to the identification, design and implementation of prioritized tax planning opportunities for the business
- Responsible for protecting value in respect of tax risks through the operation of appropriate policies, procedures and frameworks
- Maintain key relationships with stakeholders in the business so that tax plans are developed in partnership
- Lead, develop and engage project teams where appropriate in line with leadership expectations
- Generate talent and professional capability through the coaching and development of team members on the tax technical and commercial understanding of the business
- Share experience by providing coaching, and support to the wider tax team
Essential Education
Essential experience and job requirements
You'll be a Direct Tax professional acting as a senior member within your current organisation. You'll hold a tertiary degree, most likely in a Commerce or Law related discipline who may also be CPA or CA qualified. As an agile, driven and engaging professional you'll have experience working with large corporates and within complex environments driving outcomes through communication.
Other Requirements (e.g. Travel, Location)
Desirable criteria & qualifications
Relocation available
Travel required
Negligible travel
Is this a part time position?
About BP
We are a global energy business involved in every aspect of the energy system. We have 75,000 employees in 80 countries, working towards delivering light, heat and mobility to millions of people, every day. We are one of the very few companies equipped to solve some of the big complex challenges that matter for the future. We have a real contribution to make to the world's ambition of a low carbon future. Join us, and be part of what we can accomplish together.
Corporate & Functions
Closing Date


text:
The National Highway Traffic Safety Administration (NHTSA) upgraded the rear wheel drive version of Tesla Model 3 with its company's typical crash test battery, and the complete electric sedan received a rating of 5 paintings.
This ranking is consistent with the test results of NHTSA on Tesla's previous car, all of which was rewarded. Model X SUV won five stars with all categories and subcategories in 2017. Model S has 5.4 stars that seems impossible in 2013.
The subcategory of the crash test includes impact evaluation of frontal, side, and rollover. It does not include evaluation of autopilot, Tesla's aid evaluation. To date, security agencies do not implement scoring systems for these functions. The closest thing to evaluation programs like Autopilot is for companies dealing with more advanced semi-autonomous technologies. Voluntarily, so far only Ford, GM, Weimo, Startup Nullo had submitted safety reports.
But NHTSA is investigating whether Autopilot played the role of crash when the driver broke ankle in May. The agency had previously given authority to blame Joshua Brown, who used this feature when colliding with the tractor trailer in the summer of 2016 to Tesla and Autopilot. (National Transportation Safety Board has not reached agreement)
A few months after NHTSA rated Chevy Bolt, the most direct competitor, a safety assessment of Model 3 was announced. Volt has been given an overall rating of five stars, but it has just won five stars with just over half of the subcategories.
Hope you like the news:
Tesla Model 3 awarded a five-star safety assessment
#Stay Tuned For More Updates :)


text:
SERVICE TIMESSundays: 10:45 am
Centennial High School
4757 North Rd
Circle Pines, MN 55014
We have lots of free parking!
North Community Pastor, Mike Kaelber
GRACE NORTH FACEBOOK Connect with your neighbors on our Grace North Facebook Group
Grace North is the newest community of Grace Church meeting at Centennial High School in Circle Pines. We are one church, gathering in two communities. Whether you feel far from God, are new to faith, or have been in a relationship with Jesus for a long time, you are welcome here. To really grow in our relationship with God, it's essential to be in real community with other Jesus followers. So, we want to do everything we can to create fun, safe and "easy to be in" environments of connection here at Grace.
Please check out the pages throughout this website to get a feel for who we are and how you can connect here. Visiting the "I'm New" page on our website is a great place to start to learn more about who we are and what we believe.
Together, taking one step closer to loving Jesus and loving people.
North Community Pastor
Throughout my life I have had people invest in me, by challenging me to a deeper relationship with Christ. I wouldn't be who I am today without them. Some of these people impacted me through a single conversation, while
We love to laugh, have fun and discover real life through God's Word, the Bible. A typical service at Grace lasts about seventy minutes and consists of several elements: a warm welcome, inspiring live worship music, and a message from the Bible from one of the members of our teaching team. It's a very casual, come-as-you-are environment, so no need to dress up.
Plan to show up a few minutes early to enjoy a free cup of coffee before the service and have an opportunity to meet some people. We want you to feel right at home!
Our First Stop area in the lobby is a place where you can meet some of our staff and volunteers and ask any questions you may have about Grace. We're looking forward to meeting you and helping you get connected as soon as you're ready.
Every Sunday we transform space at the school into a colorful, fun and safe environment for your children through grade 5. A computerized check-in system allows us to get in touch with you quickly if your child needs you. Want to find out more about GraceKids?
Students (grades 6 to 12) are encouraged to attend the worship service with their parents. They are also welcome to be part of Grace Student Ministries on Wednesday nights from 6:30 to 8:30 at our Roseville location. Here's where you can find out more.


text:
Your business faces many security risks these days. Unfortunately, stolen paper records are the not only threat to your data security. Criminals also steal discarded hard drives and other electronic meads in order to gain access to sensitive personal and business information.
Simply deleting data from your hard drives is not enough to protect them from being compromised. The most effective solution for protecting your digital information is the use of a professional media destruction service.
Hard Drive Destruction Protects Your Data
Thieves cannot retrieve data from your electronic media when you use Shred Guard's NAID AAA Certified hard drive and media destruction service. We protect your privacy through the use of:
Our state-of-the-art shredding equipment uses a steady crushing motion to punch through the chassis of each hard drive to destroy its internal platter, and render it inoperable and the data unrecoverable.
It is easy to verify the destruction process because we do all of our shredding on-site at your facility in our mobile shredding vehicles. Additionally, we provide you with a Certificate of Destruction when we complete your electronic media destruction project.
Shred Guard helps you destroy your confidential data with confidence. For more information about our media destruction services, please contact us by phone or complete the form on this page.


text:
Working at home
Create your personal symbol! Many people believe that only key corporations have recognizable logos, but this can be simply just not correct. Readers who stop by your internet site often will arrive to recognize and trust your emblem, and observing it on a further website or forum might remind them to visit. Viewers are your customers, and also you want to give them a little something familiar to return to.
The real key to producing good material will be to solve a dilemma to the reader. Discover a problem related using your area of interest and several techniques to unravel it. At the time you've got carried out the research simply put it collectively in the quick publish. Be pleasant in tone and also to the point. Your audience will arrive back for more.
Preserve your articles very easily obtainable no matter how long ago that it absolutely was prepared. Include trackbacks within your recent content to similar products from a long time in the past. This can assist you to both equally in search rankings and in click on throughs. If you're able to get another person intrigued more than enough to keep clicking by means of to different web pages on your site, you happen to be on your own method to owning a repeat visitor. Go to below: http://ultimatecursos.com.br/
Create article content and evaluations on the items that you are applying. You may have to appear proficient and satisfied that has a products on your viewers to wish to try it out. Giving in depth assessments or data relating to the product or service will help your audience have an understanding of the merchandise and need to click via.
Provide insightful, helpful and valuable information on your web site. Just take some beyond regular time to jot down up some informative evaluations then make use of the affiliate hyperlink to show the readers where by to visit get that exact merchandise. Throwing random links which have very little to complete with precisely what is on the website is sure to result in a quick exit because of the readers. Take a look at listed here: http://ultimatecursos.com.br/
You should not bogus it! For those who are likely to compose an evaluation of the product or service, examination it very first. Readers should be able to inform you are lying and if they can not, they are going to absolutely know when their ordeals are nothing like yours. Should you are dishonest, you can reduce the shoppers you labored so challenging to gain.
Utilize a pull to drag people in. Audience want to have their curiosity contented. In case you question a matter or make them speculate from the title of one's site, audience is going to be attracted to finding out exactly what the response is. Use this to your gain by dragging visitors into your web site, and hopefully they can adhere around to your exciting written content.
Remember to go to our website: http://ultimatecursos.com.br/


text:
Quick, easy and stress free slow grilled beef ribs that can be cooked and ready in under 2 hours. Perfect for all the family.
Serves 4
Prep 20 minutes
Cook time 1 h 30 minutes
Ready in 1 h 50 minutes
|· 4 pounds/1.8 kg beef short ribs
· for marinade:
· 1/2 cup/120 mL dry red wine
· 1/4 cup/60 mL soy sauce
|· 2 tablespoons/30 mL vegetable oil
· 2-3 cloves garlic, minced
· 1 teaspoon/5 mL dried thyme
· 1/2 teaspoon/2.5 salt
· 1/2 teaspoon/2.5 mL black pepper
In a medium plastic or glass bowl combine marinade ingredients. Mix well. Place the ribs in a shallow, non-metal baking dish and pour the marinade over them. Make sure the ribs are evenly coated. Cover with plastic wrap and refrigerate for a 2-6 hours.
Preheat grill and prepare for indirect grilling. When the grill is hot remove ribs from marinade and place on grill to cook indirectly. Discard marinade.
Cook for 1 to 1 1/2 hours turning every 15 minutes. Once ribs reach an internal temperature of 165 degrees (make sure to check several), remove from heat and serve. If you are so inclined, you can apply barbecue sauce during the last 15 minutes of cook time. Make sure to keep a close eye on the ribs, so that they do not burn.
Recipe Sourced from The SpruceEats
More Beef Recipes Here


text:
By Susan Blumenthal, MD, MPA and Ladan Fakory, MPA
With as many as 500,000 spectators and more than 10,500 athletes expected to attend the Olympic and Paralympic Games in Rio de Janeiro this summer, the world is racing to contain the mysterious, mosquito-borne Zika virus that has infected more than 120,000 people in Brazil since 2015. Several athletes and journalists have already said they will not attend this year's games. It's no wonder why: The disease causes microcephaly, a rare birth defect – never before associated with a viral infection – that causes babies of some pregnant women, infected with the virus, to be born with underdeveloped heads and brains. More than 1,300 cases of confirmed microcephaly in infants born to mothers who were infected with Zika during pregnancy have been reported in Brazil. Another 3,300 cases are still being investigated. Earlier this week, the Centers for Disease Control and Prevention (CDC) revealed that there are currently 234 pregnant women in the continental United States who are infected with the Zika virus and 189 in the US territories. The CDC reports that three babies have been born in America with birth defects caused by the virus, and another three pregnancies ended in either miscarriage or termination where there were signs of Zika-related abnormalities. All of the cases reported in the U.S. were connected to travel to regions of the world with outbreaks of the mosquito-borne virus.
The likelihood of a woman who was infected with the Zika virus during pregnancy giving birth to a baby with microcephaly is estimated to be as high as 13 percent. With approximately 4 out of 5 people infected with Zika having no symptoms, researchers studying the outbreak in Colombia reported last week that even pregnant women who don't show symptoms of infection are at risk of having babies with microcephaly. Additionally, physicians are concerned that infants might experience other neurological damage and developmental problems as a result of in-utero Zika infection that could affect a child's vision, hearing, cognition, and behavior. These alarming consequences led the World Health Organization (WHO) to issue interim guidance on June 7, 2016 that those living in Zika affected areas "be correctly informed and oriented to consider delaying pregnancy." Adults have experienced an autoimmune disease associated with the virus called Guillain-Barré syndrome that can result in paralysis. To date, several people infected with Zika have died.
On June 14, 2016, the WHO convened the third meeting of the International Health Regulations Emergency Committee to assess the impact of Zika and concluded that the risks posed by the virus are not high enough to postpone or move the 2016 Olympic Games. Pregnant women are advised not to travel to Brazil for the Olympics. The Committee's experts also restated the importance of the availability and use of insect repellent and condoms for athletes and visitors, intensified vector control measures, enhanced surveillance, risk communication, and clinical care. The WHO strongly advises people returning from areas with local Zika infection transmission to adopt safer sex practices, consider abstinence, and delay pregnancy for at least 8 weeks upon return, and 6 months if the male partner has shown any symptoms of infection.
With no vaccine or treatment for Zika, or indeed much knowledge about who is most at risk for complications of the disease currently available, 150 bioethicists, scientists, physicians, and public health professionals wrote a letter to the World Health Organization (WHO) on May 27, 2016 urging that the Olympic Games be postponed or relocated. They worry that some athletes and spectators will become infected while in Rio and then carry the virus back to their home countries, further spreading the disease worldwide. This would be of particular concern for countries where the virus is not endemic and for developing nations that do not have adequate health infrastructure to prevent the disease's spread and where mosquito vectors exist for transmission. They argue that major sporting events have been moved before; for example, the Women's World Soccer Cup in 2002 was relocated from China over concerns about the swine flu. In their letter to the WHO, these experts believe that just a few infected travelers in several countries or continents could result in a "full blown global health disaster." Additionally, there is a "perfect storm" brewing in Rio with financial and politic problems that could impact the course of the Zika epidemic there.
However, other scientists suggest that the number of Zika cases will decline during August-September, the winter months in Brazil, when the Olympics are scheduled to occur. However, recent seasonal patterns for diseases, like dengue, carried by the same Aedes aegypti mosquito, suggest that while the Zika outbreak might well ebb during the summer, it will most likely not disappear entirely. Furthermore, given the effects of climate change, some mosquitos are thriving in winter months that are warmer than in previous years. Another concern expressed by some experts is that the Asian strain of Zika (believed to be introduced to Brazil in 2013 and subsequently to other South and Central American countries) appears to have higher rates of serious complications, including microcephaly and Guillain-Barré syndrome, as compared to the African strain. More research is needed on these observations, and on other aspects of the disease. In terms of concerns about Zika transmission in America, it is interesting to note that despite more than a million cases of chikungunya (a viral disease carried by the same mosquitos as Zika) reported in the Caribbean, Latin and South America over the past several years, this disease did not spread throughout the United States. Additionally, a report of last week's number of cases suggests that the chikungunya's incidence has declined dramatically over the past months except in certain countries including Brazil, Colombia, the Honduras, and Bolivia. This is likely due to "herd" immunity—a large number of people have been infected already-- and a drought in the Caribbean. Thus, given there have only been small localized outbreaks of dengue and chikungunya in the United States (mainly in the Southwest including Florida and Texas), despite larger outbreaks elsewhere in the world, this pattern of small outbreaks will probably be the case with Zika in America as well. However, we really don't know for sure.
But what we do know is this: Tens of thousands of tourists packed into outdoor stadiums and event venues at the Olympics this summer is worrisome to athletes and spectators when there is a mosquito-borne disease at large. To address these concerns, the WHO and the Centers for Disease Control and Prevention (CDC) recently issued statements to reassure the public that changing or cancelling the location of the 2016 Olympics would not significantly alter the international spread of the disease. These organizations note that many people already travel to and from Brazil every month for many reasons, and that, since 2007, mosquito-borne transmission of Zika has been reported in more than 60 countries and territories worldwide and in 40 countries in the Americas alone. Ten countries have reported person-to-person transmission of the virus. According to Dr. Tom Frieden, Director of the CDC, "The Olympics would represent less than 0.25 percent of all travel to Zika affected areas." He states, "So even if the Olympics were called off, we'd still be left with 99.75 percent of the risk of Zika continuing to spread."
While the WHO and CDC statements provide general reassurance that the Rio Olympic games this summer do not pose an increased risk of Zika transmission given the sheer volume of travel that already occurs to and from Brazil, the actual risk of further Zika spread as a result of the Games is difficult to accurately predict. To address this issue, several epidemiologists have developed a mathematical model based on estimating how many mosquitos will be in Rio during the Games to determine the chance that spectators and athletes could be infected during this three-week period. Their preliminary model suggests a 1 in 31,000 chance of contracting Zika during the Olympics in Rio – that means 16 cases– as compared to a 1 in 11 million chance of dying in a plane crash or a 1 in 9,100 chance of being killed in a car accident in the United States this year. According to this mathematical model, keeping the Games in Rio would not change the course of the epidemic globally. Similarly, a San Paulo-based research group confirms this prediction, estimating that attendance at the Rio Olympics would result in a maximum of 15 Zika infections among the visitors. Given the public's fears about contracting Zika at the Games, Rio's Olympic organizers convened a briefing on June 7th, with the city's chief medical expert emphasizing these results from the mathematical modeling --that Zika is expected to affect only an estimated 1.8 people per 1 million tourists –15 people--during the three weeks of the Olympic competition. According to the European Centers for Disease Control and Prevention, attendees are much more likely to experience food poisoning during the Olympics in Brazil than to become infected with Zika. A number of scientists also believe that if an international event were to have furthered the spread of Zika, it would have been the Carnival festivities in February when Rio was hosting a million visitors and when mosquito proliferation was at its peak.
However, other experts believe there are limitations to the statistical mathematical approach described above. It should be remembered that this public health issue is not just about the number of cases, but also about whether infected people traveling back to their countries might trigger a new outbreak somewhere in the world, especially in slum-dwelling environments where WHO and CDC precautions and preventive strategies may not be implemented .
Additionally some ethicists and scientific experts argue that core principles animating the Olympic Games provide a counter argument to convening the Olympic competition in Rio this summer under these circumstances. An article published in the Harvard Public Health Review quotes the International Olympic Committee statement: "Olympism seeks to create … social responsibility and respect for universal fundamental ethical principles." If any Olympic athletes or spectators attending the 2016 games in Rio become infected with the virus and return to their respective countries and then transmit the disease to others, this would represent a violation of the Games' principles of social responsibility, which if interpreted broadly, would include not spreading an infectious disease. These concerns are echoed by bioethicist, Dr. Arthur Caplan, who writes that the WHO is "betting on the weather, responsible behavior by visitors, adequate mosquito control and a low sexual-transmission rate by returning visitors. All are gambles."
What is clear is that the ongoing spread and impact of the Zika virus is yet another wake-up call after the Ebola outbreak last year that infectious diseases pose significant threats to human health and international security. On February 1, 2016, WHO declared that the Zika outbreak was a Public Health Emergency of International Concern. Experts estimate that 2.17 billion people are living in tropical and sub-tropical regions, where Zika-carrying mosquitos can thrive. According to the WHO, as many as 4 million Zika cases are predicted in the Western hemisphere alone by the end of 2016. There is increasing concern that the disease could affect people living in the United States during the upcoming summer season. As of June 15, 2016, there have been 756 Zika cases reported in 46 U.S. states and Washington D.C. (none of which have been locally transmitted), and 1,440 cases reported in 3 U.S. territories, with 1,386 cases in Puerto Rico alone. The virus is a particular threat to Puerto Rico, where local transmission is occurring. It has been estimated that as many as 25% of people residing there may become infected with the Zika virus.
Thus far, local transmission of Zika has not occurred in the continental U.S. states. However, U.S. officials are gearing up to deploy "rapid response teams" of 10-15 experts to any community that has even one case of infection to help local authorities monitor cases, perform laboratory tests, and increase mosquito control efforts. States considered most vulnerable to Zika transmission during the summer months include Alabama, Arizona, California, Florida, Georgia, Hawaii, Louisiana, Mississippi, and Texas. On June 14, 2016, an Interim CDC Zika Response Plan, a step-by step emergency plan, was released, and on June 13, 2016, applications were due to CDC to apply for $85 million in funds to fight Zika in communities. If local transmission of the virus occurs in North America, the Aedes aegypti mosquito would likely be the primary vector in urban areas and southern U.S. states along the Gulf Coast, whereas the Aedes albopictus mosquito could play a role in transmission due to its distribution as far as New England and the lower Great Lakes. The global transportation of cargo and travel by people are largely responsible for introducing these mosquito vectors across countries and continents. In addition, environmental changes including urbanization, dam construction, deforestation, and rising temperatures linked to climate change have increased the number of locations around the world in which these mosquitos can thrive. That's why intensified efforts are underway to fight this virus including the development of vaccines to prevent further transmission and rapid early diagnostic methods. An experimental vaccine for the Zika virus will begin human testing for safety and appropriate dosage in the coming weeks after receiving approval from the FDA. The vaccine works by priming the immune system to fight Zika by introducing genetically-engineered material that mimics the virus in the body. In animal studies of this vaccine, antibodies were developed that attacked the virus. However, confirming the safety and effectiveness of any Zika vaccine will require large scale studies that would not be completed until 2017. Additionally, in recent months, several rapid diagnostic Zika blood and urine tests have also received emergency use authorization from the FDA.
As with the Ebola outbreak last year, the ongoing Zika pandemic underscores the importance of prevention and public health preparedness in mitigating the spread of infectious diseases and promoting global health. Since 1976, as many as three new infectious diseases have emerged each year: HIV/AIDS, SARS, Lyme's Disease, H1N1 influenza, Ebola, MERS, and West Nile encephalitis are just some of the most serious and headline grabbing ones. As of 2014, more than 346 infectious diseases were linked to 16% of all human deaths worldwide.
Given the urgency of Zika as a public health threat, the Obama Administration requested $1.8 billion to fight the disease. Alarmingly, the Republican majority in Congress did not approve its request. Consequently, on April 6, 2016, the Administration redirected $589 million in unspent funding to combat Ebola to fight the Zika epidemic. Escalating public concerns about the potential spread of the disease to America has resulted in Congress currently debating what level of additional funding should be provided to fight this disease. By investing in research (which has still not occurred at an adequate level) toward the discovery of a vaccine and antiviral treatments as well as building emergency preparedness infrastructure, human suffering and the long-term costs of caring for children with microcephaly, and people with Guillain-Barré syndrome and other health damaging complications of infection with the virus can be averted. Additionally, the results of increased knowledge about Zika should pay dual dividends to help fight other infectious diseases as well.
People can continue to debate where the Olympics should be held this summer, but Zika is a global problem regardless of where athletes go for gold this summer, a public health threat that requires more resources and research to eradicate. On May 6, 2016, UN Secretary-General Ban Ki-moon announced the establishment of a targeted "UN Zika Response Multi-Partner Trust Fund," which will provide a rapid, flexible, and accountable platform to support a coordinated response to support the global Zika Strategic Response Framework and Joint Operations Plan in consultation with UN agencies, partners, and international public health experts. Donors will contribute to a central "fund" and an Advisory Committee will direct monies to priority activities and programs. This is an important step forward, but we believe and have proposed that this Zika Fund be expanded and established as a "Global Health Security Fund," a permanent entity to support the international response to all emerging infectious disease threats now and in the future.
On May 20, the World Bank launched an innovative $500 million insurance fund to address some of this need. The new Pandemic Emergency Financing Facility (PEF) will help in the fight against pandemics in poor countries. In the event of an outbreak, the facility will rapidly release monies to an affected nation and qualified global first responder organizations. The fund creates the first insurance market for pandemic risk. The Bank estimates that if the PEF had existed in mid-2014 as the Ebola outbreak was rapidly spreading in West Africa, as much as $100 million could have been mobilized by July to limit the spread of the epidemic. Instead, resources did not begin flowing until about three months later permitting Ebola cases to increase by ten-fold during that period of time.
Last month, at the G-7 meeting in Tokyo, pandemic planning was a prominent issue with the group's endorsement of the Global Health Security Agenda. The G-7 has reached out to more than 70 countries to assist them in implementation of the WHO International Health Regulations. However, only about one-third of nations globally have the resources and public health infrastructure to respond effectively to public health emergencies like Zika creating vulnerabilities worldwide. That is why pandemic planning and response is essential with coordination of efforts, robust early detection methods, disease surveillance, and reporting systems, as well as investments in new vaccines, treatments, and prevention strategies. Community education and mobilization is critical, as well as creating a Pandemic Response Corps of trained health professionals, lab technicians, and other health providers, who can be rapidly mobilized when an outbreak occurs anywhere in the world.
The bottom line: focusing global attention on where the Olympics should be run this summer while failing to commit the necessary time, money, and action to combat the Zika virus is not a winning strategy. Throughout history, infectious diseases have killed more people than wars, making them powerful international enemies and decisive shapers of history. They remain clear and present dangers to human health, economic development, and national security in an interconnected 21 century world, and we must remain vigilant against them. That is why a medical marathon is needed, making significant investments now to strengthen the scientific knowledge base, developing new global health technologies including rapid testing methods that can be deployed in combination with proven public health practices, as well as strengthening health systems with a coordinated national and international response. These efforts will enhance our ability to fight Zika and other emerging infectious disease threats more swiftly and effectively, moving from peril to progress in the years ahead. Intensifying public concern about the spread of the Zika virus worldwide underscores that microbes are ticking time bombs against the health of humanity and failure to fight them is not an option.
Susan Blumenthal, M.D., M.P.A. is the Public Health Editor of The Huffington Post. She is a Senior Fellow in Health Policy at New America, Senior Policy and Medical Advisor at amfAR (The Foundation for AIDS Research), and a Clinical Professor at Tufts and Georgetown University Schools of Medicine. Dr. Blumenthal served for more than 20 years in senior health leadership positions in the federal government in the Administrations of four U.S. presidents including as Assistant Surgeon General of the United States, the first Deputy Assistant Secretary of Women's Health, and as Senior Global Health Advisor in the U.S. Department of Health and Human Services, where she focused on global health diplomacy. She also served as a White House advisor on health. Dr. Blumenthal was involved in the U.S. response to bioterrorism with the anthrax attacks in 2001. She has provided pioneering leadership in applying information technology to health, establishing one of the first health websites in the government (womenshealth.gov) and the "Missiles to Mammogram" Initiative that transferred CIA, DOD and NASA imaging technology to improve the early detection of breast and other cancers. Prior to these positions, Dr. Blumenthal was Chief of the Behavioral Medicine and Basic Prevention Research Branch, Head of the Suicide Research Unit, Coordinator of Project Depression, and Chair of the Health and Behavior Coordinating Committee at the National Institutes of Health (NIH). She has chaired many national and global commissions and conferences and is the author of numerous scientific publications. Admiral Blumenthal has received many awards including honorary doctorates and has been decorated with the highest medals of the U.S. Public Health Service for her pioneering leadership and significant contributions to advancing health in the United States and worldwide. Named by the New York Times, the National Library of Medicine and the Medical Herald as one of the most influential women in medicine, Dr. Blumenthal was named the Health Leader of the Year by the Commissioned Officers Association and as a Rock Star of Science by the Geoffrey Beene Foundation.
Ladan Fakory, M.P.A. is a Health Policy Fellow at New America in Washington, D.C. She received her Master's in Public Administration from Harvard University, John F. Kennedy School of Government. She has 11 years of health experience across the U.S. Department of State, U.S. Agency for International Development (USAID), and Walter Reed Army Institute of Research (WRAIR), where she focused on HIV/AIDS prevention, maternal and child health policy, global health diplomacy, and the Millennium Development Goals (MDGs).
An earlier version of this article was published in the New America Newsletter on June 9, 2016.