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A coiled-coil interaction mediates cauliflower mosaic virus cell-to-cell movement
Livia Stavolone, Maria Elena Villani, Denis Leclerc, and Thomas Hohn
PNAS April 26, 2005 102 (17) 6219-6224; https://doi.org/10.1073/pnas.0407731102
Livia Stavolone
Maria Elena Villani
Denis Leclerc
Thomas Hohn
Edited by Patricia C. Zambryski, University of California, Berkeley, CA, and approved March 7, 2005 (received for review October 20, 2004)
The function of the virion-associated protein (VAP) of cauliflower mosaic virus (CaMV) has long been only poorly understood. VAP is associated with the virion but is dispensable for virus morphogenesis and replication. It mediates virus transmission by aphids through simultaneous interaction with both the aphid transmission factor and the virion. However, although insect transmission is not fundamental to CaMV survival, VAP is indispensable for spreading the virus infection within the host plant. We used a GST pull-down technique to demonstrate that VAP interacts with the viral movement protein through coiled-coil domains and surface plasmon resonance to measure the interaction kinetics. We mapped the movement protein coiled-coil to the C terminus of the protein and proved that it self-assembles as a trimer. Immunogold labeling/electron microscopy revealed that the VAP and viral movement protein colocalize on CaMV particles within plasmodesmata. These results highlight the multifunctional potential of the VAP protein conferred by its efficient coiled-coil interaction system and show a plant virus possessing a surface-exposed protein (VAP) mediating viral entry into host cells.
movement protein
virion-associated protein
Biacore
Cauliflower mosaic virus (CaMV) is the type member of the Caulimoviridae family (1). Its double-stranded DNA genome contains six major ORFs encoding the functional and structural viral proteins (2). Of these ORFs, only the capsid protein (and/or its polyprotein precursor GAG), the polymerase polyprotein, and the transactivator/viroplasmin are essential for virus replication (2, 3), whereas the movement protein (MP) and the virion-associated protein (VAP) are additionally required for induction of plant disease (4-6).
VAP, a rather small protein (15 kDa) with a bipolar functional structure (Fig. 1), associates with capsid protein in virion shells through its C-terminal region (7, 8). Its N-terminal region has a high α-helical content and harbors a coiled-coil motif assembling into a parallel tetramer with a rod-like structure (9). The tetramer is the stable form of VAP found in infected plants (10). VAP also contains a nonsequence-specific nucleic acid-binding domain mapped at the C-terminal proline-rich domain between amino acids 112-126 (11).
Genome organization of CaMV ORFs I-VII, functional maps of MP, ATF, and VAP. CP, capsid precursor polyprotein; POL, polymerase polyprotein (reverse transcriptase/RNase H and aspartic protease); TAV, transactivator/viroplasmin. MP, VAP, and ATF are depicted by boxes with amino acid numbers indicated. Hatched boxes in MP and VAP represent the nucleic acid-binding domains, and the black boxes represent the coiled coils. tfd, tubule-forming domain; ffd, focus-forming domain; vbd, virus-binding domain.
In both plant and animal systems, coiled-coil domains mediate not only protein homo-oligomerization but also interaction between distinct proteins (12). Based on its bipolar structure, we speculated that VAP, anchored by its C-terminal domain in the virus capsid, could act as the virion "arm," mediating important CaMV functions through interaction with other proteins. Indeed, the VAP coiled-coil cooperates with the aphid transmission factor (ATF), the 18-kDa nonstructural protein encoded by CaMV ORF II, which oligomerizes in a parallel-oriented trimer (Fig. 1 and refs. 13 and 14). The N-terminal region of ATF interacts with the inner lining of the aphid stylet, exposing its C-terminal α-helical domains for the coiled-coil-dependent association with VAP (14), which itself is bound to virions (7). Thus, VAP mediates CaMV transmission by aphids through coiled-coil interactions with ATF.
Transmission by aphids is an important function but is not required for virus infection of a single plant host. The finding that plant infection through mechanical inoculation of a VAP-defective CaMV strain is unsuccessful (6), whereas virus replication in a single cell is independent of this protein (5), suggests that VAP not only mediates aphid transmission but could also be involved in virus movement.
Two different mechanisms of virus cell-to-cell movement have been recognized and studied extensively. In one, MP is associated with viral RNA in a nucleoprotein complex that moves to the neighboring cell (15); in the other, the protein modifies plasmodesmata by insertion of tubular structures, allowing entire virus particles to be transported (16). CaMV MP (encoded by ORF I; Fig. 1 and ref. 4) features both RNA binding activity (17, 18) and formation of tubules containing virus particles that protrude from infected protoplasts (19). However, a number of lines of evidence confirm that CaMV moves as virions through tubules, and the biological relevance of the RNA binding activity remains to be determined. MP accumulates in foci at the periphery of transfected protoplasts, and the tubules it forms extend beyond into the medium (19, 20). The functional domains of MP involved in both these functions have been determined and mapped (Fig. 1 and refs. 21 and 22). CaMV MP is the only component of tubules (23); nevertheless, it colocalizes with the Arabidopsis thaliana MPI7 protein in the cell periphery foci (24), suggesting the possible involvement of host factors in virus movement.
Here, we investigate the involvement of VAP in movement of CaMV. We show that MP, like ATF, oligomerizes as a trimer through a C-terminally located coiled-coil domain and, through the same domain, interacts with the coiled-coil of the VAP located on the virion surface. The MP-VAP interaction occurs within plasmodesmata-containing virus particles.
Protein Expression and Purification. CaMV ORF I (encoding MP), ORF III (encoding VAP), and derivatives thereof were amplified by PCR from CaMV clone pCa37 (25) by using gene-specific oligonucleotides containing appropriate restriction endonuclease recognition sequences at the precise extremity of each gene. PCR products were cloned into the BamHI-EcoRI sites of pGEX-2TK (Amersham Pharmacia), which contains a thrombin recognition site, as in-frame fusions with the GST domain, and were overexpressed in Escherichia coli BL21. ORF III was also cloned in the NcoI-BamHI sites of pET-3d (Novagen) and expressed in the same cells. MP and VAP expression were induced by growing the cell cultures for 3 h with 0.4 and 0.2 mM IPTG, respectively.
For Biacore surface plasmon resonance assays, GST-tagged proteins were purified on GSTrap FF columns (Amersham Pharmacia). While bound to the column, proteins were cleaved from GST by a 16-h incubation with thrombin protease (Amersham Pharmacia) at 22°C with gentle rotation. Excess protease was removed, allowing cleaved protein to flow through a HiTrap Benzamidine FF column (Amersham Pharmacia). All purification steps were performed according to the manufacturers' instructions.
GST Pull-Down Assay. The pull-down assay was performed essentially as described in ref. 26 with some modifications. The challenging proteins were produced by overexpression in E. coli BL21. Supernatants obtained after bacterial lysis and centrifugation were incubated for 1 h with proteins fused to GST already bound to beads. Aliquots (5-10 μl) of the bound fraction were separated by SDS/PAGE, and proteins detected by Western blotting by using anti-VAP-coiled-coil antibody (7).
Cross-Linking. Chemical cross-linking of purified MP was performed in 20 mM sodium phosphate buffer/150 mM NaCl (pH 7) at a concentration of 0.3 mg/ml with 1 or 5 mM of Bis 2-sulfosuccinimidooxycarbonyloxyethylsulfone (Pierce) for 2 h at 4°C. The reaction was quenched for 20 min with 50 mM Tris buffer, pH 7.5. Products were analyzed by Western blotting with a polyclonal antibody obtained from a rabbit immunized with MP coiled-coil peptide. Oxidative disulfide cross-linking of the MP coiled-coil peptide was performed as described in ref. 9.
Size Exclusion Chromatography. The MP coiled-coil peptide was separated on SUPERDEX 200 resin prepacked in a PC 3.2/30 column according to the manufacturer's instructions (Amersham Pharmacia), collecting 0.5-ml aliquots of the free-flow rate. The column was calibrated with chymotrypsinogen A (25 kDa), cytochrome c (12.4 kDa), and VAP pep(wt) (4,149 Da; ref. 9).
CaMV Purification. CaMV strain CabbS was propagated in Brassica rapa cv. "Just Right" and virus purified as described by Gardner and Shepherd (27) with an additional centrifugation step through a 10-40% sucrose gradient. Purified virions were isolated by high-speed centrifugation (45,000 × g for 1 h) of the band collected from the gradient diluted 1:1 in water.
Surface Plasmon Resonance (SPR). SPR is a label-free technology for monitoring biomolecular interactions as they occur. It measures mass changes induced by association or dissociation between an immobilized analyte and a binding partner. A Biacore X optical biosensor system (Biacore, Neuchâtel, Switzerland) was used to measure the kinetics of molecular interactions, expressed in response units. One thousand response units of purified MP, diluted in 10 mM sodium acetate (pH 4.0), were immobilized on the dextran matrix of a CM5 sensor chip by using an amine coupling kit according to the manufacturer's instructions. Real-time interaction analysis was performed by injecting different concentrations of VAP (ranging from 10 to 160 μg/ml) in the absence or presence of virus onto the MP-coated chip. Binding of VAP to an excess of CaMV virions (150 μg/ml) was performed by overnight incubation at 4°C with gentle shaking. All binding experiments were carried out in Hepes-buffered saline (10 mM Hepes/0.15 M NaCl/3 mM EDTA/0.005% surfactant P20), at 25°C and at a flow rate of 20 μg/ml. Ten millimolar HCl pulses were used to regenerate the chip surface. Kinetic rate constants (Kon, Koff, and Kd; see Fig. 6) were calculated on the basis of a single-site model by using Biacore biaevaluation 3.0 software.
Biacore SPR analysis of the interaction among MP, VAP, and virions. Sensograms show real-time interaction of immobilized MP with increasing concentrations (10-160 μg/ml, as indicated adjacent to traces) of VAP (A) and VAP associated with CaMV virions (B). A steep upward slope defines the onset of binding partner injection (VAP in A and VAP-virion complex in B). The adjacent ascending regions illustrate association between test partners. The abrupt downward slopes correspond to the end of the injection period and define the start of binding partner dissociation (descending curve). (The steepest parts of the graphs, almost parallel to the y axis, reflect only changes in buffer conditions and do not represent interaction dynamics). Kon, association constant; Koff dissociation constant; Kd = Koff/Kon, equilibrium dissociation constant.
Electron Microscopy.Brassica rapa cv. "Just Right" leaf tissue systemically infected with CaMV was fixed in 4% glutaraldehyde in Pipes (pH 7.4) for 3 h at room temperature under mild vacuum. After extensive washing in Pipes, samples were dehydrated in a graded ethanol series and embedded in LR White resin. Ultra-thin sections of these samples were used for double immunogold labeling as described in ref. 28 with some modifications (see Supporting Methods and Fig. 8, which are published as supporting information on the PNAS web site, for detailed methods). Sections were stained with 2% aqueous uranyl acetate for 15 min and lead citrate for 5 min before viewing with a Zeiss EM 910 transmission electron microscope operated at 80 kV.
Protein Computational Analysis.seqweb, the web-based version of the Wisconsin Package Version 10.2 (Genetics Computer Group, Madison, WI) was used for all protein sequence analyses. Coiled-coils were predicted with coilscan (29, 30).
A Coiled-Coil Domain Predicted at the C Terminus of MP Assembles as a Trimer. Inspection of the CaMV MP sequence revealed the presence of a heptad repeat at the C terminus of the protein. In the same region, computer analysis predicts a coiled-coil domain with high probability (Fig. 2A and ref. 29). The ability of MP to form oligomeric structures was investigated by chemical cross-linking of the purified protein. Western blotting analysis with anti-MP antibody showed three forms of MP, corresponding in size to protein monomers, dimers, and trimers (Fig. 2B). Moreover, trimerization of a peptide corresponding to the coiled-coil domain alone demonstrated the latter's involvement in MP oligomerization (Fig. 2C). For this purpose, a peptide of 2,385 Da, encompassing the predicted coiled-coil of MP [coilMP(wt), residues 307-327], was synthesized. A derivative peptide, coilMP, with the addition of the sequence GSCECKQN to the N terminus of coilMP(wt) (Fig. 2D), was constructed to allow us to distinguish whether the peptide assembles to form trimers in parallel or antiparallel orientation. In vitro cysteine oxidative cross-linking would allow formation of S-S bridges between parallel-oriented monomers as the cysteine residues (in bold) would be located in close proximity in the oligomeric state, whereas no cross-linking of three monomers together would occur if the peptides assembled in antiparallel orientation. The highest band detected by SDS/PAGE under denaturing conditions without reducing agent showed that coilMP assembles into a parallelly oriented trimer (Fig. 2C). Chromatography confirmed the trimer as the most stable level of oligomerization of the peptide. CoilMP(wt) peptide loaded on a gel filtration column (Superdex 200) together with the standard molecular mass markers and coilVAP(wt), a synthetic peptide corresponding to the VAP coiled coil that elutes as a tetramer (9), eluted as a single peak with a mass <12 kDa (Fig. 2E), corresponding to the predicted trimer mass (7,155 Da). The absence of MP coiled-coil monomers and dimers suggests that all of the coilMP(wt) exists as a single population of trimers in solution, thus we interpret the presence of monomers and dimers in Fig. 2 B and C to be due to incomplete cross-linking. CoilMP and coilVAP eluted individually, and no heteromers of the two peptides were detected under these conditions.
Analysis of CaMV MP structure. (A) Computer prediction of coiled-coil domains within the MP sequence. The entire MP protein is represented on the x axis and coiled-coil probability (%) on the y axis. (B) Chemical cross-linking of MP analyzed by SDS/PAGE and detected by immunoblot analysis with anti-MP coiled-coil antibody. Positions of the monomer and the cross-linked dimers and trimers (1, 2, and 3, respectively) are indicated [CaMV MP (37 kDa) migrates anomalously on SDS/PAGE, ref. 21]. The product in the left lane was not treated with Bis 2-sulfosuccinimidooxycarbonyloxyethylsulfone. (C) Disulfide cross-linking of CoilMP analyzed by oxidative SDS/PAGE and silver staining. Positions of the peptide monomer and cross-linked dimers and trimers (1 = 3,235 Da, 2 = 6,470 Da, and 3 = 9,705 Da) are indicated. The product in the left lane was not treated with oxidized glutathione. (D) Synthetic peptide CoilMP, corresponding to the C-terminal α-helical domain of MP (nonnative amino acids in italics), with the seven amino acid unit (depicted as a-g) of the heptad repeat structure illustrated above; hydrophobic residues of the α-helix occupy positions a and d (highlighted in bold). (E) Separation of reduced CoilMP(wt) by gel filtration on Superdex 200 (Amersham Pharmacia). Standard molecular mass markers are chymotrypsinogen A (25 kDa, •) and cytochrome c (12.4 kDa, ▴). The elution peaks correspond to CoilMP(wt) peptide trimer (7,155 Da, solid black line), and CoilVAP(wt) peptide tetramer (16,596 Da, dashed black line). Protein concentration was determined by monitoring absorbance at 280 nm of each 0.5-ml elution fraction.
CaMV MP Interacts with VAP Through Its C-Terminal Region. A possible MP-VAP interaction was investigated further in GST pull-down experiments. A GST-MP fusion (Fig. 3A) was overexpressed and purified by using Glutathione-Sepharose 4B beads. The GST-MP-bead complex obtained was incubated with partially purified VAP produced in an E. coli expression system. After extensive washing, proteins bound to the beads were eluted and separated on SDS/PAGE. Western immunoblotting with antibody raised against VAP was used to reveal the presence of the protein in the eluted protein-bead complex. Using this technique, VAP was indeed found to associate with full-length MP (Fig. 3B, lane 1).
Mapping of MP domains involved in the interaction with VAP. (A)MP and MP deletion mutants are depicted by boxes of distinct colors with the construct names indicating the corresponding amino acid positions. White boxes represent GST, expressed in fusion with the proteins; black boxes represent the coiled-coil domain. Dotted lines replace deleted amino acids. The white vertical line in construct 295-327ΔIDL represents the three amino acid deletion. (B) Interaction of VAP with MP and its mutants fused to GST and with GST alone (GST, lanes 5 and 11). Proteins were separated by SDS/PAGE and detected by Western blotting with antibodies against VAP. Positions of molecular mass marker proteins are on the left of the gels. Lanes 6 and 12, VAP alone.
To map the MP domains involved in the interaction with VAP, N-terminal, central, and C-terminal fragments of MP were expressed in fusion to the C terminus of GST (Fig. 3A) and used in GST-pull down experiments. VAP associated with the C-terminal region of MP but not with other MP fragments or GST alone (Fig. 3B, lanes 1-5). Analysis of two further mutants of the GST-C-terminal fragment of MP [GST-MP (251-294) and GST-MP (295-327)] revealed the last 32 C-terminal residues of MP to be sufficient to support the interaction with VAP (Fig. 3B, lanes 8 and 9).
Thomas and Maule (31) determined the MP sequences required for virus infectivity by three-amino acid deletion scanning. All mutants unable to mediate systemic infection carried a deletion within the tubule-forming domain except one in which the mutation was located exactly in the region required for VAP binding. When this deletion, ΔIDL (amino acids 314-316), was introduced into GST-MP (295-327), VAP-binding capacity was lost (Fig. 3B, lane 10). This result suggests that the lack of infectivity of the MP(ΔIDL) mutant is due to the inability of MP to interact with VAP and, consequently, to mediate virus cell-to-cell movement.
CaMV MP and VAP Interact Through Their Coiled-Coil Domains. The results obtained so far indicate a physical association of VAP with the coiled-coil domain of MP. To determine the region of VAP involved in this interaction, two deletion mutants of VAP, corresponding to the N-terminal coiled-coil (residues 1-58) and the C-terminal region (residues 58-129) were produced and cloned in the pET3a vector (Fig. 4A). GST-MP (295-327) was overexpressed, bound to beads and challenged with both VAP mutants. Only the N-terminal VAP mutant precipitated with the complex, thus confirming that MP and VAP associate through their coiled-coil domains (Fig. 4B).
Mapping of VAP domains involved in the interaction with MP. (A) VAP and VAP deletion mutants are depicted by boxes, with the construct names indicating the corresponding amino acid positions. The coiled-coil region is shaded black. (B) Lanes 1-3, interaction of GST-MP (295-327) with VAP and its mutants. Lane 4, VAP alone. Proteins were separated by SDS/PAGE and detected by Western blotting with antibodies against VAP. The position of the 14.3-kDa marker (lysozyme) is on the left of the gel.
Caulimovirus MP and VAP Colocalize in Plasmodesmata. The subcellular localization of MP and VAP, each expressed in fusion with the green fluorescent protein (GFP), has been investigated previously in protoplasts. MP preferentially aggregates in foci at the plasma membrane and in tubular structures emerging from the cell surface (20), whereas VAP forms large aggregates within the cell cytoplasm (L.S., unpublished data).
We investigated the localization of these proteins in CaMV-infected Brassica rapa leaf tissue by double immunogold electron microscopy with rabbit antibodies against MP and VAP and, as secondary antibodies, goat anti-rabbit immunoglobulins conjugated to 18- and 6-nm gold particles, respectively. MP was found to be associated with the cell membrane and concentrated within plasmodesmata (Fig. 5 A-C). Notably, MP is more abundant in the inner part of the plasmodesma (Fig. 5B), where the MP-formed tubules are believed to traverse the cell wall (19). No MP aggregates similar to those observed in infected insect cells (21) were found in infected turnip leaf tissue, suggesting that formation of such aggregates could involve other components that are specific to, and/or more abundant in, the insect system.
VAP and MP colocalize in modified plasmodesmata. Electron micrographs of ultrathin sections of infected tissue showing two neighboring cells. Immunogold labeling of VAP (6-nm gold particles) and MP (18-nm gold particles) shows the localization of the two proteins. (A) Two plasmodesmata (P) traversing the cell wall (CW) are indicated; one is modified and contains virus particles (V). (B) Longitudinal section of a modified plasmodesma containing CaMV particles (V) showing MP distribution. (C) Magnified image of the modified plasmodesma in A; Inset is enlarged to show MP and VAP localization on a single virion. (D) CaMV inclusion body containing virus particles. Gold particles (6-nm) label VAP. (Scale bars: 100 nm.)
VAP was never observed in isolation, being invariably associated with CaMV virions, located both within and outside the virus inclusion bodies (Fig. 5 and ref. 7). We observed that 18-nm and 6-nm gold particles were closely associated with each other only on virus particles within plasmodesmata (Fig. 5 A and C). This colocalization of MP and VAP was not seen in any other cell compartment, including virus particles close to plasmodesmata (Fig. 5 B and C). This finding suggests that the interaction between the two proteins most probably occurs at the entrance to, or during movement of virions through, the plasmodesmal cytoplasmic sleeve.
Affinity Between MP and VAP Depends on the Presence of CaMV Virions. The electron microscopic observations indicate that interaction between MP and VAP in vivo occurs only in the presence of CaMV virions in plasmodesmata. In addition, MP and VAP were never found associated when overexpressed in fusion with fluorescent proteins or when immunostained after fixation, either in protoplasts or in leaf tissue upon bombardment.
We used SPR to further investigate the affinity and dynamics of the MP-VAP association and the involvement of CaMV virions in the interaction.
Consistent with our in vitro GST pull-down experiments, VAP was trapped by MP immobilized on the sensor chip, albeit in limited amounts and through a rather fast dissociating interaction (Fig. 6A). The anomalous curves illustrating VAP dissociation from MP (Fig. 6A, irregular descending slopes) correlate with the multimeric state of VAP in solution and did not allow calculation of the corresponding kinetic constants.
When a VAP-CaMV virion complex was injected onto immobilized MP, a high-affinity interaction occurred between MP and VAP (Kd ≈ 50 nM, Fig. 6B; CaMV virions alone did not bind measurably to MP, data not shown). Indeed, when trapped in the virion, more VAP bound to MP and subsequent dissociation was much slower [compare ascending (association) and descending (dissociation) parts of the graphs between Fig. 6 A and B]. Thus, when associated with the virion, VAP interacts strongly and stably with MP. This result suggests that, under in vivo conditions, it is the VAP-virion complex, and not VAP alone, that interacts with MP. These results may account for the lack of interaction observed between MP and VAP when overexpressed in vivo.
A Coiled-Coil Domain Is Conserved in the MPs of the Caulimoviridae. If, as we propose, the interaction between MP and VAP through coiled coils is essential for CaMV movement and spread of infection in the plant, we would expect this feature to be conserved in other members of the Caulimoviridae, a virus group characterized by well conserved genome architecture and functional strategies (2). Indeed, we previously demonstrated that tetramerization of VAP is a conserved feature of all members of the group (32). Amino acid sequence analysis, supported by specific bioinformatic tools (29), revealed that a coiled-coil motif is indeed conserved, albeit not always perfectly, at the MP C terminus of those caulimoviruses having a defined MP and 3′ of the conserved intercellular transport domain (18) of all other members of the Caulimoviridae family (Fig. 7A), highlighting the importance of both domain and structure in this virus family. The case of petunia vein clearing virus is slightly different in that it encodes a single viral polyprotein containing three closely spaced coiled-coil domains. The presumptive MP, containing only the first coiled-coil domain (Fig. 7A), can trimerize (L.S., unpublished data). This finding suggests that tetramerization of a presumptive petunia vein clearing virus VAP (32) would be directed by the other two coiled coils. However, further studies on petunia vein clearing virus posttranslational regulation are required to clarify whether VAP and MP functions are indeed conserved in this virus.
Prediction of Caulimoviridae MP coiled-coil domains and illustration of VAP-mediated CaMV functions. (A) MP C-terminal domains of representative members of the six genera of the Caulimoviridae family are aligned, the numbers representing amino acid position in each MP or polyprotein. The seven positions of the heptad repeat are labeled a-g above the alignment. Positions a and d, occupied by hydrophobic residues that are part of the oligomer core, are highlighted; the other positions are predominantly solvent-exposed polar residues. Sequences are aligned according to their phylogenetic relationships (shown on the left). Phylogeny was inferred from the intercellular transport movement domains located upstream of the coiled coils (corresponding to residues 127-157 in CaMV MP). Cassava vein mosaic virus, CsVMV; commelina yellow mottle virus, ComYMV; rice tungro bacilli-form virus, RTBV; soybean chlorotic mottle virus, SbCMV; petunia vein clearing virus, PVCV. (B) Schematic overview of VAP interactions mediating CaMV functions during the infection cycle. In the aphid stylet, VAP associated with the virion interacts with ATF to mediate aphid transmission. In the MP-modified plasmodesma, the VAP-virion complex interacts with MP, allowing movement to a neighboring cell. Also shown are MP and ATF trimers and VAP tetramers, which are known to occur in planta, although the function of these forms remains unknown. Interactions between the coiled coils of MP and VAP and ATF and VAP are shown only schematically and are not intended to depict the degree of coiled-coil multimerization.
CaMV VAP and MP are both essential for the spread of infection in a host plant, but both are dispensable for replication in a single cell. In infected cells, VAP is always associated with the virion but is not an essential component of the viral capsid structure. The fact that VAP binds to virus particles when added externally (14) and is dispensable for genome packaging (5) indicates that VAP is not a scaffolding protein needed for virus assembly but is rather a late addition during the assembly process, suggesting a role in virus spread in the infected host plant.
Our results show that CaMV MP harbors a C-terminal coiled-coil domain, assembled as a parallel trimer, that interacts with the VAP N-terminal coiled coil, supporting the view that VAP is involved in virus movement. Although both proteins contain a nucleic acid binding domain, neither DNA nor RNA mediate the VAP-MP interaction, as shown by the efficient association of VAP and MP deletion mutants containing only the coiled coils and lacking the nucleic acid binding domain.
We have previously speculated that VAP mediates important CaMV functions through its bipolar structure, with one end anchored to the virion and the other, the coiled-coil domain, exposed for interaction with other proteins (32). However, despite the presence of VAP tetramers in planta (10), it remained hard to see how tetramers could fit into an icosahedral virion characterized by 2-, 3-, 5-, and 6-fold, but not 4-fold, symmetry (33). In fact, cryo-electron microscopy experiments have shown that binding to virions triggers a structural shift, changing VAP tetramers into trimeric oligomers emerging from the 3-fold symmetrical axes of the capsid (34). The coiled-coil domains of these parallel trimers become rearranged to form antiparallel dimers with a monomer from a trimer in close proximity, thus creating a network of VAP coiled-coil dimers covering the surface of the virion (34). If this behavior can be extrapolated to ATF and MP, all three of these coiled coil-containing proteins could be reoriented from parallel trimers or tetramers to associate as heterologous, antiparallel coiled-coil complexes that target the virion to different functions of the infectious cycle as depicted in Fig. 7B. In this model, we speculate that the function of the parallel coiled-coil tetrameric form of VAP found in planta could be to impede heterologous interactions until the proper partner is available. In addition, other transport-related functions possibly involving the tetrameric form of VAP, such as vesicular transport (also mediated by coiled-coil interactions; ref. 35), could be envisaged. Such strategies necessarily require strict regulation, the mechanism of which would be fundamental to targeting of different aspects of the infection cycle in CaMV. Thus, investigations are underway to understand how VAP opts between ATF and MP.
The coiled coil we mapped in MP shares three important features with the ATF coiled coil: they are both located in the C-terminal part of the proteins, assemble as parallel trimers, and interact with the amphipathic N-terminal coiled coil of VAP that is exposed on the external surface of the virus particle.
CaMV MP forms tubular structures predicted to cross the cell wall by passing through plasmodesmata (23). Structural analysis, performed by epitope tagging, located the N- and C-termini of MP on the protein surface on both the outer and inner surface of the tubules (31). The model suggested by our results is consistent with this prediction: MP coiled coils exposed inside the tubule are accessible for interaction with virus particles through VAP (Fig. 7B).
Tubules can form on the surface of CaMV-infected protoplasts upon transient expression of MP. Thus, no other virus protein is required for their formation, although host factors might be involved in folding and support of the tubule structure in the host tissue. How tubules are formed is currently unknown. However, the coiled-coil domain is the only part of MP not required for their assembly (20, 31), thus excluding a direct involvement of the MP trimer in tubule structure formation. Nevertheless, a possible role in stabilization or in virion spacing within the tubule cannot be ruled out, especially considering that, as for VAP in the virion shell, a structural shift of the trimer may appropriately reorient the oligomer. In support of such tempting speculations, the coiled-coil region is undoubtedly essential for virus spread within the host (31). Indeed, we demonstrated that a mutation in the MP coiled coil that hinders virus movement (31) also inhibits association with the VAP coiled coil. Interactions between such domains are highly specific as well as highly dependent on proper protein folding (12). Whereas the precise mechanism of virus particle movement through tubules is still unknown, our results indicate an essential role for VAP in mediating the association between virion and tubule through the VAP-MP interaction.
Our EM observations revealed that in infected cells, MP and VAP apparently associate only in the presence of virus particles passing through plasmodesmata. Biacore SPR investigations on the dynamics of the interaction showed that, as for VAP-ATF (36), the affinity of MP for VAP is highly compromised in the absence of virions. In particular, when the two proteins interact alone, faster dissociation is observed. If the MP-VAP association is indeed essential for virion movement through tubules, it is not surprising that the presence of virus particles favors maintenance of this interaction in vivo. Taken together, these observations suggest that the interaction between MP and VAP cannot occur in the cytoplasm, and that the two proteins probably move independently to the plasma membrane, with VAP comigrating with virions. MP can assume different conformations depending on its subcellular localization (31), and these conformations most probably correlate with different MP oligomeric structures that mediate distinct functions, e.g., tubule formation, binding/nonbinding to VAP, etc. Whether the virion-VAP complex interacts with MP before or after the latter forms tubule structures remains to be clarified. However, it is possible that interaction with VAP triggers the MP conformational modification necessary to allow virus movement through the tubules.
CaMV virions invade new cells only by passing through MP-modified plasmodesmata. Through the interaction with MP, VAP protruding from the virion allows virus penetration into neighboring cells. Therefore, like animal viruses, this plant virus has developed a system of cell entry mediated by surface-located viral proteins, albeit adapted to specific conditions in the host organism. Such comparisons justify parallel and complementary research lines also in terms of the evolutionary relationships linking plant and animal viruses (1).
VAP is a multifunctional protein, a tool produced and used by the virus to regulate various functional steps in the viral infection cycle. In a wider context, VAP-ATF/MP is a simple system that can be used to study general regulation of coiled-coil interactions. The α-helical coiled coil is one of the most common oligomerization motifs in proteins (12), and the CaMV model system could be of great benefit in furthering our understanding of a general strategy used by proteins to mediate biological function.
We thank Helge Rixner for expert technical assistance and advice with EM, Franz Fischer for peptide synthesis, Donato Boscia for producing anti-MP coiled-coil antibody, Mike Rothnie for graphical work, Helen Rothnie and Marcello Russo for critical reading of the manuscript, and Angiola Desiderio, Etienne Herzog, and Kappei Kobayashi for many valuable discussions.
↵† To whom correspondence should be addressed. E-mail: l.stavolone{at}ba.ivv.cnr.it.
↵¶ Present address: University of Basel, Botanical Institute, Plant Health Unit, CH-4056 Basel, Switzerland.
Author contributions: L.S. designed research; L.S., M.E.V., and D.L. performed research; M.E.V. contributed new reagents/analytic tools; L.S., M.E.V., D.L., and T.H. analyzed data; and L.S. and T.H. wrote the paper.
This paper was submitted directly (Track II) to the PNAS office.
Abbreviations: ATF, aphid transmission factor; CaMV, cauliflower mosaic virus; MP, movement protein; SPR, surface plasmon resonance; VAP, virion-associated protein.
Copyright © 2005, The National Academy of Sciences
Rothnie, H. M., Chapdelaine, Y. & Hohn, T. (1994) Adv. Virus Res. 44, 1-67.pmid:7817872
Hohn, T. & Fütterer, J. (1997) Crit. Rev. Plant Sci. 16, 133-161.
Kobayashi, K. & Hohn, T. (2003) J. Virol. 77, 8577-8583.pmid:12857928
Thomas, C. L., Perbal, C. & Maule, A. J. (1993) Virology 192, 415-421.pmid:8421892
Kobayashi, K., Tsuge, S., Stavolone, L. & Hohn, T. (2002) J. Virol. 76, 9457-9464.pmid:12186927
Jacquot, E., Geldreich, A., Keller, M. & Yot, P. (1998) Virology 242, 395-402.pmid:9514961
Leclerc, D., Stavolone, L., Meier, E., Guerra-Peraza, O., Herzog, E. & Hohn, T. (2001) Virus Genes 22, 159-165.pmid:11324752
Leh, V., Jacquot, E., Geldreich, A., Haas, M., Blanc, S., Keller, M. & Yot, P. (2001) J. Virol. 75, 100-106.pmid:11119578
Leclerc, D., Burri, L., Kajava, A. V., Mougeot, J. L., Hess, D., Lustig, A., Kleemann, G. & Hohn, T. (1998) J. Biol. Chem. 273, 29015-29021.pmid:9786907
Tsuge, S., Kobayashi, K., Nakayashiki, H., Mise, K. & Furusawa, I. (1999) Microbiol. Immunol. 43, 773-780.pmid:10524795
Mougeot, J. L., Guidasci, T., Wurch, T., Lebeurier, G. & Mesnard, J. M. (1993) Proc. Natl. Acad. Sci. USA 90, 1470-1473.pmid:8434006
Burkhard, P., Stetefeld, J. & Strelkov, S. V. (2001) Trends Cell Biol. 11, 82-88.pmid:11166216
Hebrard, E., Drucker, M., Leclerc, D., Hohn, T., Uzest, M., Froissart, R., Strub, J. M., Sanglier, S., van Dorsselaer, A., Padilla, A., et al. (2001) J. Virol. 75, 8538-8546.pmid:11507199
Leh, V., Jacquot, E., Geldreich, A., Hermann, T., Leclerc, D., Cerutti, M., Yot, P., Keller, M. & Blanc, S. (1999) EMBO J. 18, 7077-7085.pmid:10601029
Fujiwara, T., Giesman-Cookmeyer, D., Ding, B., Lommel, S. A. & Lucas, W. J. (1993) Plant Cell 5, 1783-1794.pmid:12271056
van Lent, J., Storms, M., van der, M. F., Wellink, J. & Goldbach, R. (1991) J. Gen. Virol. 72, 2615-2623.pmid:1940857
Thomas, C. L. & Maule, A. J. (1995) Virology 206, 1145-1149.pmid:7856089
Koonin, E. V., Mushegian, A. R., Ryabov, E. V. & Dolja, V. V. (1991) J. Gen. Virol. 72, 2895-2903.pmid:1684985
Perbal, M. C., Thomas, C. L. & Maule, A. J. (1993) Virology 195, 281-285.pmid:8317106
Huang, Z., Han, Y. & Howell, S. H. (2000) Virology 271, 58-64.pmid:10814570
Thomas, C. L. & Maule, A. J. (1999) J. Virol. 73, 7886-7890.pmid:10438886
Huang, Z., Han, Y. & Howell, S. H. (2001) Mol. Plant-Microbe. Interact. 14, 1026-1031.pmid:11497463
Kasteel, D. T., Perbal, M. C., Boyer, J. C., Wellink, J., Goldbach, R. W., Maule, A. J. & van Lent, J. W. (1996) J. Gen. Virol. 77, 2857-2864.pmid:8922481
Huang, Z., Andrianov, V. M., Han, Y. & Howell, S. H. (2001) Plant Mol. Biol. 47, 663-675.pmid:11725951
Lebeurier, G., Hirth, L., Hohn, T. & Hohn, B. (1980) Gene 12, 139-146.pmid:6260583
Herzog, E., Guerra-Peraza, O. & Hohn, T. (2000) J. Virol. 74, 2073-2083.pmid:10666237
Gardner, R. C. & Shepherd, R. J. (1980) Virology 106, 159-161.
Faoro, F., Tornaghi, G. & Belli, G. (1991) J. Phytopathol. 133, 297-306.
Lupas, A. (1996) Methods Enzymol. 266, 513-525.pmid:8743703
Lupas, A., Van Dyke, M. & Stock, J. (1991) Science 252, 1162-1164.pmid:2031185
Thomas, C. L. & Maule, A. J. (1995) Plant Cell 7, 561-572.pmid:7540082
Stavolone, L., Herzog, E., Leclerc, D. & Hohn, T. (2001) J. Virol. 75, 7739-7743.pmid:11462048
Cheng, R. H., Olson, N. H. & Baker, T. S. (1992) Virology 186, 655-668.pmid:1733107
Plisson, C., Uzest, M., Drucker, M., Froissart, R., Dumas, C., Conway, J., Thomas, D., Blanc, S. & Bron, P. (2005) J. Mol. Biol. 346, 267-277.pmid:15663943
Clague, M. J. (1999) Curr. Biol. 9, R258-R260.pmid:10209109
Drucker, M., Froissart, R., Hebrard, E., Uzest, M., Ravallec, M., Esperandieu, P., Mani, J. C., Pugniere, M., Roquet, F., Fereres, A. & Blanc, S. (2002) Proc. Natl. Acad. Sci. USA 99, 2422-2427.pmid:11842201
You are going to email the following A coiled-coil interaction mediates cauliflower mosaic virus cell-to-cell movement
Livia Stavolone, Maria Elena Villani, Denis Leclerc, Thomas Hohn
Proceedings of the National Academy of Sciences Apr 2005, 102 (17) 6219-6224; DOI: 10.1073/pnas.0407731102


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HomeEU politicsBoubacar Boris Diop: "We are Independent of France, but not Free of It"
Boubacar Boris Diop: "We are Independent of France, but not Free of It"
May 28, 2020 Mathew D. Rose EU politics, National Politics 0
A special weekend read. Senegalese writer Boubacar Boris Diop argues that the independence of French-speaking African countries is a pending battle
Interview by Jaume Portell Caño
Cross-posted from El Salto
Translated and Edited by BRAVE NEW EUROPE
When Boubacar Boris Diop (Dakar, 1946) was a child, Senegal was a French colony and he lived in a neighbourhood created on the fringe of the city: the Medina. In Dakar – the Senegalese capital – the white settlers in Plateau, the administration quarter, didn't want to mix with the blacks , in case the blacks infected them with any disease, but they needed them nearby so they could come to serve, clean, and care for them.
Thus was born, next to Plateau, the Medina, the place where Diop grew up and became fond of reading thanks to the stories his mother told him. Today he is one of the most prestigious writers on the African continent, thanks to novels and essays that range from the Rwandan genocide – Murambi, the Book of Bones (2709 Books, 2019) – to Africa's relations with France – Africa beyond the Mirror (2009), The Book of Secrets (Almuzara, 2015). At the time of the interview, Diop finds himself in an office presided over by a giant portrait of the writer, anthropologist, nuclear physicist and pan-Africanist politician Cheikh Anta Diop, whom he considers a model.
What did independence mean for Senegal?
We are independent, but we are not free. France continues to rule this country through what we call "Françafrique". After defeats in Vietnam and Algeria, Paris understood that it had to change its relations with the Africans. They put it this way: "We will give them their independence, they will have a flag, a president, a government, a national team. They will have all the symbols of political power, but we will continue to command through an elite who will obey us. Sixty years after independence we have no political sovereignty. Everything is decided by France, including the currency we use, the CFA franc.
The debate about the CFA has grown rapidly recently, especially among young people.
I think that for many years there has been a fear of talking about it among intellectuals: President Olympio, in Togo, was assassinated in the 1960s for trying to get out of the CFA. Joseph Pouemi, a Cameroonian economist who was very critical of the currency, was also murdered in 1984. Today young people ignore the mainstream media, and any one of them can send a message to Macron with a video, put it on YouTube, and get millions of people to watch it. It is a novelty that people are now talking so much about the CFA, and I am very happy about that, because three or four years ago it was a very marginal issue.
Many Senegalese risk their lives to come to Europe. If Boubacar Boris Diop were 18 today, would he follow that path or would he stay in Senegal?
When I was that age I had the opportunity to go to France on a scholarship from the Senegalese government, but I preferred to stay. I didn't want to leave my mother, who was having financial difficulties, alone, and I remained here because I wanted to help her. In today's context, many young people see France, Spain, or Italy as countries where you can be happy and have opportunities. Here they have nothing and they don't see any kind of future: instead of living like wild animals they choose to leave, even if it's very risky.
Do you think that this diaspora will help change African countries?
It's already happening! In the elections, the Senegalese Diaspora voted for Ousmane Sonko, a very different choice from the current president of Senegal, Macky Sall. In the Diaspora we have engineers, university professors, all these intellectuals are already helping to shape current debates. This translates into actions such as that of a woman who publicly embraced the president of Senegal in London. The Diaspora is spreading the political awareness of Africans.
In The Book of Secrets, Yacine Ndiaye, one of the protagonists confesses her greatest wish: to be white and become a baroness with European surnames. Why does she want to do this?
We don't like ourselves. If you look at it, Senegalese TV presenters are lighter-skinned. This pressure is especially aimed at women: you won't see black men trying to become Brad Pitt. We must ask ourselves how we got to this point. In the book I presented this issue as a joke, but it is really happening.
Why is the figure of Cheikh Anta Diop so important to you?
He was the first to consider that Africa was the cradle of humanity and that ancient Egypt was a black civilization. He also promoted an important idea: we had to be proud of ourselves and think independently in order to achieve real political sovereignty.
Language was a crucial part of this struggle. The French, through Senghor [a Senegalese poet who became head of state in Senegal], managed to remove him from academic life in Senegal, but he remained in touch with the people. After independence, the University of Dakar continued to function as a French university, which is why they decided that Cheikh Anta Diop could not be in contact with African students.
In Murambi, your book on the Rwandan genocide, you comment on the role of the French. What role did France play in Rwanda?
The interim government of Rwanda, which later became the perpetrator of the genocide, was established in the French embassy. The genocidaires never ceased to have the political support and weapons of the French. Everything they needed was provided by Paris, including diplomatic recognition at the United Nations.
Each chapter in Murambi presents a character who explains his vision. What led you to choose this format?
I wanted the book to reflect all the voices, including the killers. That's why it's not a normal novel that goes from A to Z, but presents different voices and situations. The first character, Michel Serumundo, works at the Kigali market, returns home and his son Jean-Pierre is not there. It's April 6, 1994, the day the genocide began. He says he'll try to find him and that's the end of the chapter. We don't know anything else. Did he find Jean-Pierre, was he killed? There's an uncertainty. Not knowing anything about what happened to so many people is precisely what happened during the Rwandan genocide.
What did you learn in Rwanda?
We don't really know Africa. Africa is not a people: what happened in Rwanda is very different from what happened in the Congo, Sierra Leone, Ethiopia or Somalia. We have to make a better connection between our past and our present to understand the social and historical dynamics of each country.
Your neighbourship, La Medina, was created so that whites would not have to live with blacks. They lived separately, but close enough so that whites could still count on the resources and services of Africans. Do you think this is a metaphor for globalisation?
We are invaded by images, companies, and Western nations. They tell us this is globalisation, but we can't go to their "neighbourhood". It is very ironic that Westerners can go to Kenya, Lesotho or South Africa without a visa. But I need a visa to go to any Western country. It's like we live in a ghetto or a big prison and they say "we need you, but please don't come".
Boubacar Boris Diop
Jaume Portell Caño
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The South Carolina African American Heritage Foundation supports the efforts of the South Carolina African American Heritage Commission to identify and promote the identification and preservation of the African American experience in SC.
Our Mission…. is to support the efforts of the South Carolina African American Heritage Commission to identify and promote the preservation of historic sites, structures, buildings, and culture of the African American experience in South Carolina, and to assist and enhance the efforts of the South Carolina Department of Archives and History.


BREAKING: The first deal of this year's Toronto Film Festival he goes to Samuel Goldwyn, which acquired the doc Diana Vreeland: The Eye Has To Travel. Goldwyn acquired North American rights and will put the film out theatrically in at least 20 markets. The film was directed by Lisa Immordino Vreeland, wife of the late fashion magnate's grandson. It got strong reaction in Venice and Telluride, and after a private screening at the Royal York Hotel last night Goldwyn, IFC, Magnolia and Sony Pictures Classics and Music Box went after it. Goldwyn took it around 4 AM. Pic was repped by Submarine's Josh Braun.


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Bumpy deficits, smoother ride: The historical evolution of budget deficits and growth rate
By Jared Bernstein and Richard Kogan Aug 18, 2015, 10:30am EDT
Share All sharing options for: Bumpy deficits, smoother ride: The historical evolution of budget deficits and growth rate
This post is part of Polyarchy, an independent blog produced by the political reform program at New America, a Washington think tank devoted to developing new ideas and new voices.
When it comes to fiscal policy and especially budget deficits, your basic DC policymaker walks around thinking, "Deficits bad, surpluses good." Virtually every Republican currently running for president — and more than a few congressional Democrats — have endorsed amending the Constitution to force Congress to balance the budget each year.
Even those who recognize that there are rare times when it makes sense for the government to temporarily spend more than it takes in tend only to defend deficits as a necessary evil.
We think this is wrong. Budget deficits, properly managed, are not only not evil — they're downright good. The long historical record shows that their use has led to smoother and stronger growth.
In essence, they've worked like shock absorbers, smoothing out bumps along the growth path. To eschew their use, as so many policymakers espouse, would mean growth that was both more volatile and somewhat slower.
Thanks to a new, long time series of historical data, we're able to show you what we mean. Consider two time periods, which we'll call BD (before deficits) and AD (after deficits; meaning after deficits started to be more commonly employed in their shock-absorber role). In this analysis, BD runs from 1870 to 1929 and AD from 1950 to 2015 (see data note).
The BD period is a deficit hawk's dream. The deficit-to-GDP ratio flatlines in the figure through boom and bust alike, averaging a slight surplus (!) of 0.4 percent over the period. Over the AD period, deficits average -2.2 percent of GDP.
Importantly, deficits are also a lot jumpier in the latter versus the former period. The standard deviation — a measure of dispersion around the average — is 0.5 percent BD and many times that much, 2.4 percent, in the AD period.
So deficits were both a lot larger and a lot more varied in the latter period.
What about growth? Real per-person income growth is faster in the AD period, though not too much: 2.0 percent annually versus 1.8 percent in the BD period. Still, this extra growth adds up over time: With AD growth rates, the economy doubles every 35 years, but with the slower BD growth rates, it takes an additional four years to double.
And just eyeballing the figures, you can see how much jumpier income growth was in the earlier BD period; recessions were far more frequent and far more severe. The standard deviation of income growth was 5.0 percent in the BD period and less than half that — 2.3 percent — in the AD period.
In other words, the shock absorber of deficit spending is one reason the growth ride has been smoother post-1950, and very likely has contributed to faster overall growth, as well, since it is easier to maintain speed on smoother pavement.
How does it work? What we're calling "bumps in the road" are contractions in private sector economic activity, caused by one sort of shock or another, like the bursting of a housing bubble or the sharp disruption of a key input, like oil. To offset these shocks, the government borrows from the future to spend more today, which is just what the economy needs during bad times, when ordinary citizens are losing income (and jobs) and investors have few useful opportunities anyway. When the private sector is back up and running at full strength, that's the time for deficits to fall or turn to surplus.
To be sure, using deficits as shock absorbers is by no means the sole explanation for the smoother and stronger growth in the AD period. Another candidate is increased globalization, since the increased flows of goods and capital helps avoid volatile supply disruptions. The Federal Reserve also has gotten better at macro-management over time.
Of course, the fact that deficits can be good if wisely managed does not mean that deficits are always wisely managed. For instance, it's problematic to see rising ratios of debt to GDP in strong economies. As Keynes himself said, "The boom, not the slump, is the right time for austerity…"
Unfortunately, a glance at fiscal policy around the globe suggests we are in the midst of relearning this lessons. Greece is the poster child, but most European countries that went the austerity route following the Great Recession are still mired in slow growth and high unemployment. (Real GDP in the eurozone has been growing around 1 percent annually, and unemployment is 11 percent.) In other words, there are a lot of hapless passengers in other countries getting an awfully slow and bumpy ride.
Data note: It's important to leave large wars out of the picture, as they create unusual fiscal dynamics, so the BD period starts after the Civil War and leaves out 1918-'19, and the AD period starts after World War II. But the data goes all the way back to 1792, and different endpoints yield similar results.
The growth measures are real economic growth per person. Accounting for population growth deals with the fact that the population has grown faster or more slowly during different periods in history. Economic growth is measured over calendar years, deficits over fiscal years.


It's Wednesday and I am back again with another #AppyHolidays post! This week I am featuring the ultimate holiday appetizer on the blog: Sausage Rolls. Everyone loves these meaty appetizers, especially around the holidays. Not only do they taste delicious, they are super easy to make- even from scratch! Simply assemble some store bought puff pastry with ground chicken and turkey sausage and a few other festive ingredients and you'll have a great holiday appetizer that will fly off the plate in seconds! Read on to see how I make them!
For the sausage filling, I like to combine both sausage meat and ground meat in this recipe. After testing the recipe a few times, I've noticed that using plain sausage meat in the recipe yields really oily/soggy Sausage Rolls… which doesn't let the puff pastry rise enough. By adding a bit of ground chicken, it absorbs some of the fat and makes the filling a bit lighter, but still super flavourful. In this recipe I like to use ground white meat chicken with spicy turkey sausages- perfect for the holidays! If you can't find spicy turkey sausages, just use some regular pork sausage… it will taste just as delicious! I just combine these two meats in a bowl and add a bit of fennel seed and mustard for extra flavour. Then, I spread the sausage filling in a long line on a sheet of puff pastry and roll them up.
At this point I like to place the sausage rolls in the freezer for a few minutes to cool off the pastry and let it firm up again! I found that by re-freezing the pastry for a few minutes allows it to puff up much more than if it was cooked from room temperature- and we need to have puffed pastry so this step is key! Once the Sausage Rolls have firmed up they are also so much easier to slice into bites. I like to cut them into 1 inch sized bites and then brush them each with a bit of egg wash (egg mixed with water or milk) so that they get nice and golden brown in the oven.
I then bake the Sausage Rolls for 20-25 minutes at 375 degrees F, or until they are nice and golden brown and the sausage is fully cooked.
To serve the Sausage Rolls, you can go super traditional by serving them with a side of mustard for dipping. But if it's the holidays, I like to serve my Sausage Rolls with a side of Cranberry Sauce. This is actually a really great way to use up any leftover cranberry sauce that you may have too!
And there you have it, another great appetizer for the holiday season! Let me know in the comments below if you have tried any of my recipes so far in my #AppyHolidays series and how they turned out!
For the full recipe for my Homemade Sausage Rolls, click here!


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Home | About Us | History & What We Do
Staats is one of the oldest awards and recognition companies in the country, spanning into three centuries. Staats was founded in 1898 by Mr. E. G. Staats, from Lacrosse, Wisconsin. The company's primary business was the manufacturing of braid and trim for band and fraternal uniforms, as well as making pennants. The transition to award ribbons occurred around 1915-1920. The original location was above what was for many years the VFW on the Southwest corner of the square in Mount Pleasant, Iowa. In 1939, the company moved to the former Willowbank School on North Lincoln Street. In the 1980's, Staats expanded further into the awards industry by adding plaques, trophies, and similar products to its line. Over the next decade, Staats continued to grow in the promotional market by adding advertising specialties and imprinted apparel.
By the spring of 2000, Staats had outgrown the old schoolhouse and moved into a brand new facility in the Crossroads Industrial Park. The new facility is set up to maximize efficiency and creates versatility to accommodate the changing needs of its clients. It's location in the Crossroads Industrial Park is its present home.
James Patten of Davenport, Iowa purchased Staats in 1965, and remains the present owner. Since that time, Staats has become one of the premier award and recognition companies in the United States. Staats supplies its products to county and state fairs, festivals, livestock shows, schools, corporations, and other organizations locally and across the entire United States, and even extending into the U.S. Virgin Islands.
"History is a relentless master. It has no present, only the past rushing into the future. To try to hold fast is to be swept aside." - John F. Kennedy
Innovation to Meet a Changing Market
The most recent generation at Staats has seen dramatic growth in the physical plant, the product and our capabilities. Automated machinery and computers enable the company to keep abreast of a widely expanding market. Today, Staats is a leader in the awards and recognition industry. Patterned rosettes, banners, plaques, acrylics, advertising specialties, and imprinted apparel have long since replaced "pennants." The continued introduction of new concepts of awards and recognition products and processes, combined with dedicated, experienced employees, and a continued focus on quality, customer service, and efficiency allows us to help our customers Leave Their Mark on their customers, business partners, and communities.
We at Staats have the privilege and opportunity to play a part in some of the greatest moments of the lives of our clients and their associates. As we move into the next generation of Staats, we are able to use our vast experience and apply it toward a prosperous future. In all that we do at Staats, we are sure to Leave Our Mark.


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Rezistența cehă în al doilea război mondial este un subiect care nu este încă cunoscut complet, în principal datorită puținătății documentelor dar și a eficienței poliției germane, care a reușit să împiedice multe acte de rezistență sau a anihilat organizațiile de luptă cehă. În primele zile ale războiului, populația cehă a participat la boicotarea transporturilor publice și au fost chemări sporadice la proteste de masă.

Analiză istorică

Istoricul Radomír V. Luža a împărțit rezistența cehă în două faze distincte. În prima fază, care a durat până în 1942, mișcarea de rezistență s-a concentrat pe crearea unor rețele de informații militare, mici acte de sabotaj și diseminarea de informații. După lichidarea acestei mișcări de rezistență, s-a manifestat o a doua fază la sfârșitul anului 1944 în cadrul unui val de insurecții populare din întreaga Europă Centrală și Răsăriteană. Această fază a fost caracterizată prin acțiuni mult mai diversificate, având sprijinul moral al avansării aliaților din vest și est. (Vedeți și: Ocupația Cehoslovaciei).

După cum afirmă istoricul ceh Vojtěch Mastný, germanii au folosit puține trupe de ocupație în Protectoratul Boemiei și Moraviei (așa cum era numită Cehia în acele vremuri) deoarece "comportarea populației nu a justificat niciodată o creștere substanțială a efectivelor înarmate". Concluzia finală a analizei sale în lucrarea Cehii sub dominația nazistă: eșecul rezistenței naționale este aceea că: "cehii nu au reușit să opună autorităților naziste o rezistență eficientă. Până în 1942, mișcarea de rezistență a fost distrusă, nejucând un rol semnificativ până la sfârșitul războiului".

Consolidarea grupurilor de rezistență: ÚVOD

Mișcarea de rezistență care a existat în primii ani de război a fost sub conducerea președintelui cehoslovac Edvard Beneš, care, împreună cu František Moravec (șeful spionajului militar), din exilul de la Londra, au coordonat activitatea de rezistență. În condițiile luptei împotriva forțelor de ocupație germană, principalele grupuri de rezistență și-au întărit rândurile în Conducerea Centrală a Rezistenței Naționale, ori ÚVOD. Această organizație a funcționat ca intermediarul clandestin principal dintre Beneš și Protectorat, care a existat de-a lungul întegului an 1941. Scopul pe termen lung al organizației era de a funcționa ca un guvern din umbră, până când țara urma să fie eliberată de sub dominația nazistă.

Cele mai importante trei grupuri de rezistență care au funcționat în cadrul au fost: Centrul Politic, (Politické ústředí, PÚ), Comitetul Petiției "Noi rămânem credincioși" (PVVZ), și Apărarea Națiunii (Obrana národa, ON). Aceste grupuri aveau o natură democratică și se opuneau celui de-al patulea grup de rezistență, cel al Partidului Comunist Cehoslovac (KSČ). Cei mai mulți membri ai celor trei mișcări erau foști ofițeri trecuți în rezervă din armata cehoslovacă. În 1941, ÚVOD și-a însușit platforma politică a grupului de stânga PVVZ, intitulat "Pentru libertate: în o nouă republică cehoslovacă". În acest document, ÚVOD și-a proclamat credința în idealurile democratice ale fostului președinte Tomáš Masaryk, cerea proclamarea unei republici cu caracteristice socialiste și îi chema pe toți exilații să țină pasul cu progresele socialiste din țară.

În plus față de asigurarea comunicațiilor dintre Londra și Praga, ÚVOD era responsabilă pentru transmiterea rapoartelor militare. A îndeplinit această sarcină prin folosirea unei stații secrete de radio, ale cărei emisiuni puteau fi recepționate de toată populația cehă. Însă de-a lungul timpului s-a dovedit că ÚVOD transmitea date inexacte, incomplete sau depășite. Uneori aceste informații erau în mod intenționat false. Beneš cerea uneori ÚVOD să dea rapoarte fals-optimiste despre situația militară, pentru a asigura creșterea moralului, sau pentru a motiva întărirea rezistenței antinaziste.

În vreme ce ÚVOD servea ca principal sprijin al lui Beneš, organizația se depărta uneori de la linia lui politică. În vara anului 1941, ÚVOD a respins propunerile pentru expulzarea parțială a germanilor din Sudeți după încheierea războiului și a insistat în schimb pentru expulzarea lor totală. ÚVOD a reușit să-și impună punctul de vedere în această privință.

ÚVOD și comuniștii (KSČ)

Relația ÚVOD cu KSČ a fost un aspect important al funcționării zilnice a rezistenței, de vreme ce relațiile sovieto-cehe au devenit o parte centrală a eforturilor lor de luptă. Declanșarea invaziei germane în URSS din iunie 1941 a marcat punctul de cotitură în relațiile sovieto-cehe. Mai înainte de invazie, "principalul obiectiv al comuniștilor era oprirea războiului imperialist" și de multe ori comuniștii manifestau simpatie pentru muncitorii Reichului. După declanșarea războiului, rezistența a început să se bizuie pe sprijinul comunist atât în Cehoslovacia cât pe cel venit de la Moscova. Într-o emisiune de la Londra transmisă în eter via ÚVOD, Beneš și-a informat concetățenii că "relațiile dintre cele două state ale noastre s-a reîntors la situația de dinainte de München și la vechea prietenie".

De vreme ce KSČ nu era parte în mod oficial a ÚVOD și își păstrase independența organizațională, Conducerea Centrală a Rezistenței Naționale i-a chemat pe comuniști la realizarea unității de acțiune a tuturor grupurilor antiafasciste. Liderii comuniști au păstrat legături de parteneriat strânse cu ÚVOD, ajutând astfel la menținerea unor bune relații sovieto-cehe. Beneš a folosit mijlocirea liderilor KSČ pentru a aranja întâlniri la Moscova în vederea extinderii parteneriatului sovieto-ceh. Există unele dovezi conform cărora ÚVOD ar fi prevenit Moscova cu privire la iminența atacului german împotriva URSS-ului în aprilie 1941. În mai 1941, Beneš a primit informații cu privire la concentrarea de trupe germane la granițele sovietice. După cum a scris el în memoriile publicate după război, Beneš ar fi transmis imediat această informație americanilor, britanicilor și sovieticilor. Soarta KSČ a fost legată strâns de cea a ÚVOD. Cele două mișcări au fost desființate după asasinarea lui Reinhard Heydrich, și nu și-au mai revenit până în 1944.

Rezistența cehă și asasinarea lui Reinhard Heydrich

Faimosul act al rezistenței cehe, asasinarea lui Reinhard Heydrich de pe 27 mai 1942 a marcat în mod paradoxal sfârșitul rezistenței cehe și a ÚVOD. În multe privințe se poate aprecia că decesul ÚVOD a fost pecetluit după numirea lui Heydrich ca Reichsprotektor al Boemiei și Moraviei în toamna anului 1941. Până la sfârșitul lui septembrie, Heydrich reușise arestarea aproape a tuturor membrilor ÚVOD și întrerupsese aproape toate legăturile dintre rezistența cehă și Londra.

Reacția naziștilor la asasinarea lui Heydrich este considerată ca fiind responsabilă de anihilarea după 1942 a oricărei mișcări clandestine antigermane cehe. Naziștii au executat o operațiune de pedepsire razând de pe suprafața pământului două sate: Lidice și Ležáky. În octombrie 1942 au fost condamnați la moarte de către tribunalele germane din Protectorat 1.331 oameni, 1.000 de evrei au fost trimiși direct de la Praga în lagărul de concentrare de la Mauthausen, alți 252 de cehi fiind trimiși la Mauthausen pentru implicarea în complotul asasinării lui Heydrich. Până la sfârșitul operațiunilor naziste, toți membrii ÚVOD au fost arestați.

R.J. Crampton scria că, după asasinarea lui Heydrich nu au mai existat acte de rezistență vizibile, iar numărul de soldați pe cap de locuitor în Protectorat era doar cu puțin diferit de cel din însăși Germania. Mastný afirmă și el că: memoria colectivă a "Heydrichiádei", cum numeau oamenii cumplitele zile care au urmat moartea tiranului, a fost o intimidare puternică a renașterii unei opozitii active. Prin moartea sa, Heydrich și-a îndeplinit ambiția sa de căpătâi – "pacificarea" Protectoratului.

Rezistența spre sfârșitul războiului

Vezi și
 Josef Mašín, membru al mișcării de rezistență de la începutul războiului
 Jan Opletal, student împușcat în timpul unei demonstrații antinaziste
 Alois Eliáš, prim-ministrul Protectoratului
 Josef František, as al aerului care a luptat împotriva Germaniei Naziste în Polonia, Franța și Anglia
 František Moravec, șeful spionajului militar cehoslovac (1937-1945)
 Insurecția din Praga (1945)
 Insurecția națională slovacă
 Rezistența austriacă
 Rezistența franceză

Bibliografie

 Eduard Beneš,. Memoirs of Dr. Eduard Benes: From Munich to New War and New Victory. Trans. By Godfrey Lias. Connecticut: Greenwood Press, 1954.
 R.J.  Crampton, Eastern Europe in the Twentieth Century—and After. Londra/New York: Routledge, 1997. ISBN 0-415-16423-0
 Radomír Luza,.  The Communist Party of Czechoslovakia and the Czech Resistance, 1939-1945 Slavic Review, vol. 28, nr.. 4, decembrie 1969.
 Radomir Luza, The Czechs Under Nazi Rule: The Failure of National Resistance, 1939-1942.  Slavic Review, vol. 34, nr. 3, septembrie 1975.
 Vojtěch Mastný, The Czechs Under Nazi Rule: The Failure of National Resistance, 1939-1942. New York: Columbia University Press, 1971. ISBN 0-231-03303-6
 W.V. Wallace, The Czechs Under Nazi Rule: The Failure of National Resistance, 1939-1942.  The English Historical Review'', vol. 88, nr. 348. , iulie 1973.

Istoria Cehoslovaciei
Cehoslovacia
Rezistența cehă

redpajama_set_name: RedPajamaWikipedia


text:
Home Caribbean Jamaica News Jamaica to lobby international community regarding marijuana industry
Jamaica to lobby international community regarding marijuana industry
KINGSTON, Jamaica, Feb 14, CMC – The Jamaica government says it plans to lobby the international community, particularly the United States, as the island develops its marijuana industry.
Industry, Commerce, Agriculture and Fisheries Minister, Audley Shaw, says this partly stems from the reluctance by local banks to fund the industry's growth, fearing international de-risking that could have serious economic implications for Jamaica.
De-risking involves measures taken to make an engagement less risky, or less likely to involve a financial loss.
"We will be using all available diplomatic resources, including our Ambassador in Washington DC, to mount a lobbying effort to the Government of the United States, primarily through its Treasury Department, to address this very important matter," Shaw told reporters following the official opening of Caribbean Producers Jamaica Limited's (CPJ) distribution centre in Montego Bay Freeport earlier this week.
Shaw said the Government's cannabis licensing arrangement, inclusive of an established Authority, to ensure that "we are operating at global standards".
"This simply means that we are, therefore, in a position where we can convince the United States and other global economies that we are prepared to operate at the standards that are expected of us," he added.
Shaw said, however, that local financial institutions' fear of de-risking remains a challenge, and places Jamaica and other Caribbean countries at a disadvantage, especially as their entities are not in a position to act independently of their American counterparts.
"All of our commercial banks in Jamaica are obliged, right now, to go through the New York system in terms of the movement of money internationally," he said, adding "we have the irony of a Jamaican scientist who gets an approval to develop a cannabis-based drug to fight leukaemia, and from no less an authority than the United States Food and Drug Administration (FDA)… [but] who had to watch as our local banks close his accounts because of this de-risking arrangement. This is why I am going to put a lot of energy into this lobbying effort".
Shaw noted that while de-risking mainly affects smaller developing countries, like Caribbean states, the bigger developed nations "have managed to figure out a way" around the issue.
He said, however, that positive signals are emanating "at the highest level" out of the United States, pointing out that key officials "are suggesting that they would be open to take a more careful look at the benefits associated with medicinal cannabis".
"We are also seeing where one of the Democratic frontrunners for next year's US Presidential contest… [is] advocating for the legalisation of marijuana at a federal level."
Shaw said he is "also encouraged by the utterances of the man who is expected to be the next US Attorney… who seems to be also taking a more liberal approach".
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text:
La théorie des anciens astronautes est une théorie pseudo-scientifique et ufologique selon laquelle plusieurs anciennes civilisations auraient été en contact avec des « visiteurs » extraterrestres venus apporter sur la Terre quelques savoirs dans les domaines de l'écriture, de l'architecture, de l'agriculture, des mathématiques, de l'astronomie et de la médecine.

Ces « êtres » hypothétiques détenteurs d'une technologie supérieure à celle de l'Homme à ces époques seraient devenus, au fil des siècles, des « dieux », ces êtres supranaturels dont parlent les anciennes mythologies et dont l'archéologie met les cultes en évidence.

La théorie est souvent attribuée à Erich von Däniken mais, si ce dernier l'a amplement popularisée en 1968, elle reprend de nombreux éléments déjà présents dans les doctrines théosophiques publiées par Helena Blavatsky dans les années 1880, ainsi que d'autres publiés en 1960 par Louis Pauwels et Jacques Bergier, en 1962 par Robert Charroux et en 1965 par Jean Sendy. Sans compter les nombreux textes littéraires relevant de la science-fiction, comme par exemple À l'aube de l'histoire, publié par Arthur C. Clarke en 1953 .

Elle s'appuie sur une interprétation littérale de textes religieux ou de découvertes insolites comme les lignes de Nazca ou encore de faux artéfacts comme les crânes de cristal.

Pour la communauté scientifique, la théorie des anciens astronautes n'a aucun fondement, car les éléments archéologiques prétendument inexpliqués ont le plus souvent une explication rationnelle déjà exposée par les archéologues. Ainsi, Jean-Pierre Adam, dans L'archéologie devant l'imposture, explique que la construction de la grande pyramide repose non pas sur un savoir étranger à l'espèce humaine mais bien sur les techniques de l'époque.

Théorie
La théorie repose sur les hypothèses suivantes :
 les civilisations antiques (égyptienne, maya, andines...) n'auraient pas possédé les connaissances nécessaires pour réaliser certaines de leurs constructions colossales (Baalbek, statues de l'île de Pâques, géoglyphes de Nazca...) ou complexes (machine d'Anticythère…) ;
 des éléments donneraient des indices d'une présence extraterrestre : certains personnages présents sur des fresques anciennes représenteraient des astronautes (Dogū au Japon), d'autres éléments représenteraient des ovnis ou des « pistes d'atterrissage » ;
 des êtres extraterrestres auraient influencé le développement des civilisations sur Terre, en enseignant aux hommes l'agriculture, l'écriture, les mathématiques ; voire en altérant l'ADN humain pour favoriser l'évolution vers une espèce plus intelligente. On rejoint ici la théorie du dessein intelligent extraterrestre, que l'on retrouve dans le mouvement raëlien avec les Elohim, ou chez Jean Sendy ou encore Roger Vigneron, selon qui la Bible évoque le peuple des Élus (Elohim terme hébreu présent dans l'Ancien Testament, qui signifie « ceux qui viennent des cieux ») venus sur Terre dans leurs roues de lumière (Weidorjes) ;
 les peuplades primitives, dépassées par le savoir de ces « visiteurs » nettement plus avancés, auraient considéré que ceux-ci étaient des dieux.

Retombées

La théorie a eu un fort retentissement médiatique. Elle n'a jamais été sérieusement considérée comme une théorie scientifique par les historiens ou les archéologues, mais elle a donné lieu à de nombreuses retombées :
journalistiques : notamment sur Erich von Däniken (1968), influencé par Robert Charroux et Jean Sendy (1963) ;
sceptiques : les éléments archéologiques prétendument inexpliqués ont le plus souvent une explication rationnelle déjà exposée par les archéologues. Par exemple, la technique de fabrication des géoglyphes de Nazca peut aujourd'hui être expliquée par des procédés strictement humains. De même, Jean-Pierre Adam, dans L'archéologie devant l'imposture, explique que la construction de la grande pyramide repose non pas sur un savoir étranger à l'espèce humaine mais bien sur les techniques de l'époque.
sculpturales : sur un montant de la porte de Ramos, à la nouvelle cathédrale de Salamanque (), on reconnaît un astronaute en apesanteur, œuvre du tailleur de pierre Miguel Romero lors de la restauration de 1992.

Selon Jason Colavito, l'écrivain de science-fiction américain Howard Phillips Lovecraft se trouve être à l'origine de l'expansion de la théorie des anciens astronautes dans la culture populaire tout au long du . Par le biais de ses récits fictionnels dépeignant des entités extraterrestres — assimilées parfois à des anciens dieux ou démons — descendues sur Terre en des temps infiniment lointains (L'Appel de Cthulhu, Les Montagnes hallucinées…). Lovecraft aurait eu une influence déterminante sur Louis Pauwels et Jacques Bergier, lesquels ont largement contribué à lancer le thème des anciens astronautes en France, influençant à leur tour Robert Charroux, Jean Sendy et, indirectement, Von Däniken, qui donna une dimension mondiale au phénomène.

Prétendues représentations d'engins volants et d'extraterrestres du passé

Préhistoire et Antiquité
D'après certains auteurs, des représentations étranges visibles dans quelques grottes ornées, telles celle d'Altamira en Espagne ou celle de Cougnac en France, seraient des représentations d'ovnis. De même, des statuettes ou des peintures (comme les fresques du Tassili, en Algérie) ressembleraient étrangement à certaines représentations d'extraterrestres du , preuve, selon certains courants ufologiques, de l'ancienneté du phénomène.

Certaines de ces apparitions étranges peuvent avoir été des phénomènes astronomiques (comme des comètes ou des météores) ou optiques atmosphériques. L'analyse de ces faits passés est dénommée couramment rétro-ufologie. En voici quelques exemples :

 une description remontant au règne du pharaon Thoutmôsis III vers 1450  fait état de multiples « cercles de feu plus brillants que le Soleil » d'environ  d'envergure, qui seraient apparus durant de nombreux jours. Ils ont finalement disparu après « être montés haut dans le ciel » ;
 dans la Bible, le prophète Ézéchiel décrit une vision qu'on appelle le char divin : quatre êtres, munis chacun de quatre ailes, pilotent des sortes de roues flamboyantes, capables de se déplacer comme l'éclair, changeant de direction brusquement, sans se retourner ; leurs jantes sont garnies d'yeux tout autour (comme des hublots ?) ; au-dessus de leurs têtes, quelque chose comme des dômes de cristal.

 l'auteur romain Julius Obsequens écrit, en 99 , que « dans Tarquinia, pendant le coucher du Soleil, un objet rond comme un globe a pris son chemin dans le ciel d'ouest en est ».

Moyen Âge et Renaissance

À ces époques, il est surtout question de phénomènes occultes, chez des théoriciens comme Agrippa de Netessheim ou Paracelse. L'influence de la religion est réelle puisque les phénomènes célestes sont considérés comme des avertissements divins ou des expressions maléfiques imputables aux sorciers et sorcières :

 au Japon, dans la nuit du , le général Yoritsume et son armée observent près de Kyoto des sphères de lumière non identifiées, aux mouvements erratiques. Ses conseillers lui disent de « ne pas s'inquiéter car c'était simplement le vent qui faisait osciller les étoiles ».

 gravure sur bois par Hans Glaser (1561), Nuremberg. Un feuillet imprimé rapporte que le , le ciel de la ville de Nuremberg est parcouru par une multitude d'objets décrits comme étant engagés dans une bataille. On y mentionne que de petits globes et disques sortaient de grands cylindres. Ces observations sont alors interprétées comme des prodiges surnaturels, des anges et autres présages religieux. Les historiens y voient la fusion imagée de plusieurs événements historiques et phénomènes naturels non reliés entre eux et embellis par une interprétation religieuse et une transmission orale. En météorologie, on y voit la représentation artistique d'un effet de halo.

De nos jours, ces témoignages sont parfois interprétés comme l'équivalent ancien de rapports d'ovnis modernes. Pour les cas les plus souvent cités, une explication simple est fournie par les historiens de l'art :
 les « cosmonautes » de la fresque du monastère de Visoki Dečani au Kosovo (1350) sont des représentations symboliques du Soleil et de la Lune comme on en trouve dans l'art byzantin religieux de cette époque ;
 l'« ovni » du tableau de Mainardi (Madonna col Bambino e San Giovannino), qui traverse les cieux en pleine Nativité, est en réalité la représentation symbolique de l'archange Gabriel ;
 l'objet en forme de soucoupe volante sur le tableau de Paolo Uccello, la Thébaïde, est un chapeau de cardinal ;
 la fameuse pièce de 1680 censée commémorer un passage d'ovni au-dessus du ciel de France, est en fait un jeton de jeu sur lequel est dessinée une roue de la fortune.

La théorie des anciens astronautes dans la culture

De nombreuses œuvres littéraires, cinématographiques, télévisuelles et autres se sont inspirées de la théorie des anciens astronautes.

Littérature
 L'Appel de Cthulhu (The Call of Cthulhu), nouvelle fantastique de l'écrivain américain Howard Phillips Lovecraft, publiée en février 1928 dans le magazine Weird Tales. De par le monde, des sectes d'adorateurs vénèrent les Grands Anciens, des « dieux » venus des étoiles il y a des millions d'années et aujourd'hui confinés ou en sommeil dans des régions reculées de la terre ou dans les profondeurs terrestres ou océaniques par leurs vainqueurs, des dieux plus anciens encore.
 Le Testament de la Vierge (2009), roman de Anton Parks.

Bande dessinée
 Luc Orient (1967), série de Greg et Eddy Paape, publiée dans le Journal de Tintin. Si les voyageurs extra-terrestres n'ont guère eu d'influence sur l'évolution des humains (en dehors de quelques récits légendaires), ceux-ci, en revanche, interviennent dans l'histoire des extra-terrestres.
 Vol 714 pour Sydney (1968), par Hergé,  des Aventures de Tintin, où l'on trouve un personnage représentant Jacques Bergier, également en faveur de visites extraterrestres à visées « pédagogiques ».
 Les Éternels (The Eternals), personnages de comic-book créés par Jack Kirby en 1976.
 Thorgal, série franco-belge (depuis 1977) créée par Jean Van Hamme au scénario et Grzegorz Rosiński au dessin.
 Aquablue, série de science-fiction française, plus particulièrement les tomes 5 « Projet Atalanta », 12 « Retour aux sources » et 13 « Septentrion ».

Cinéma
 1968 : 2001, l'Odyssée de l'Espace, de Stanley Kubrick (1968), inspiré par le romancier Arthur C. Clarke, évoque l'existence d'un ancien contact entre les premiers hominidés et des extra-terrestres, qui seraient à l'origine des progrès de l'Humanité.
 1989 : Un dieu rebelle, réalisé par Peter Fleischmann, adapté du roman de science-fiction des frères Arcadi et Boris Strougatski, Il est difficile d'être un dieu.
 1994 : Stargate, la porte des étoiles, réalisé par Roland Emmerich. Des scientifiques découvrent une machine complexe ouvrant un passage vers une planète désertique dont les habitants sont sous la coupe d'un tyran extraterrestre, Râ, qui se fait passer pour Dieu de l'Égypte antique.
 1997 : Le Cinquième Élément, réalisé par Luc Besson.
 2004 : Alien vs. Predator, réalisé par  Paul W. S. Anderson. Les Prédators auraient appris aux humains à bâtir des temples et des pyramides.
 2007/2009 : dans Transformers et Transformers 2, réalisés par Michael Bay, il est question de pyramides d'origine extraterrestre.
 2008 : Indiana Jones et le Royaume du crâne de cristal, réalisé par Steven Spielberg. Le célèbre archéologue est à la recherche des mythiques crânes de cristal façonnés par une civilisation extraterrestre.
 2015 : Jupiter : Le Destin de l'univers, réalisé par Les Wachowski. Le film évoque en partie la théorie de David Icke.
 2017 : The Recall, réalisé par Mauro Borrelli. Dans ce film, l'ancien soldat, interprété par Wesley Snipes, fait allusion implicitement à la théorie des anciens astronautes.
 2021 : Les Éternels, réalisé par Chloé Zhao. Adaptation des comics éponymes, le film présente des extraterrestres baptisés les Éternels, liés à des entités cosmiques, qui ont accompagné et subtilement encouragé l'évolution humaine sur Terre durant ces cinq mille dernières années.

Télévision
 L'univers Stargate : Stargate SG-1, Stargate Infinity, Stargate Atlantis et Stargate Universe.
 Star Trek, la Série Originale, Saison 2 Episode 2 Pauvre Apollon
 Il était une fois… l'Espace (1982, Procidis), série télévisée d'animation française, notamment les épisodes 5, 7, 13 et 18.
 Battlestar Galactica, série télévisée anglo-canado-américaine, de Ronald D. Moore.
 Métal Hurlant Chronicles, série télévisée française, notamment l'épisode Seconde Chance.
 Alien Theory (2010–2014), série documentaire télévisée américaine de Kevin Burns, présentant la théorie des anciens astronautes en six saisons.
 Ulysse 31, notamment l'épisode 15 : La Deuxieme Arche : il est fait références aux lignes de Nazca et, implicitement, à la théorie des anciens astronautes.
 OVNI(s), fait référence à des Hommes-Lune, des anciens astronautes, possédant des technologies avancées et les transmettant aux Terriens de nos jours.

Ouvrages propagateurs de la théorie
 Louis Pauwels et Jacques Bergier, Le Matin des magiciens, introduction au réalisme fantastique, Paris, Gallimard, 1960.
 Robert Charroux, Histoire inconnue des hommes depuis cent mille ans, Robert Laffont, 1962.
 Erich von Däniken, Présence des extra-terrestres (titre originel : Erinnerungen an die Zukunft / Souvenir du futur), 1968.
 Jean Sendy, Ces Dieux qui firent le Ciel et la Terre, Le Roman de la Bible, Robert Laffont, 1969.
 Erich von Däniken, L'or des Dieux, J'ai lu , « collection L'Aventure mystérieuse », 1974.
 Robert Charroux, L'énigme des Andes, Robert Laffont, 1974.
 Erich von Däniken, Vers un retour aux étoiles, J'ai lu , « collection L'Aventure mystérieuse », 1975.
 Christiane Piens, Les OVNIs du passé, Marabout, 1977.

Notes et références

Annexes

Bibliographie critique
 Henri Broch,"Le cosmonaute maya de Palenque" : Le Paranormal, Les documents, chp 1. 1985, Seuil (coll. Points-Science, 2001) p29-43. Du même : "Au cœur de la terre, ou les mystères de l'archéologie" : Au cœur de l'extraordinaire, chp  p61-123, Ed. L'horizon chimérique, 1991 (éditions Book-e-book, coll. « Zététique, 2005). »
  , The Cult of Alien Gods : H.P. Lovecraft and extraterrestrial pop culture, Prometheus Books, 2005, .
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Articles connexes

Liens externes
 Art et ovnis ? non merci, seulement l'art, dossier de Diego Cuoghi.
 Les ovnis du passé : L'Histoire manipulée par l'ufomanie, Cercle zététique
  Page du Skeptic's dictionary sur les anciens astronautes
  « Erich von Daniken's » Chariots of the Gods ?" : Science or Charlatanism?", un article de Robert Sheaffer.

Ufologie
Sociologie de l'imaginaire
Pseudohistoire


Steeped in rich heritage and cultural dynamism, Yangon is an enchanting melting pot of East-meets-West. From the legendary 2,600 year-old Shwedagon Pagoda to the famous Bogyoke Aung San Market, satiate your wanderlust and explore all that this charming Asian city has to offer.
Located near the Yangon city centre, PARKROYAL Yangon in Myanmar is the ideal starting point for travellers who want the best of both worlds: a luxurious stay that lies within easy reach of the city's finest attractions with our friendly PARKROYAL people always ready to assist you.


Here is a wedding album from a beautiful wedding at the Museum of the Shenandoah Valley & The George Washington Hotel in Winchester, VA.
Hey I was at that wedding! You all did a great job at capturing the day. It was so pretty!


Total Population 1911 to 1961: Population fell from 2,973 in 1911 to 2,869 in 1961.
Age Structure, in 5-year bands, from 1911 to 1961: In 1911, only 7.13 per cent of the population were aged 65 and over, but by 1961 17.85 per cent were.
Infant Mortality Rate 1911 to 1973: The highest rate recorded was 158.73 infant deaths per thousand live births in 1923, and the lowest was 0. in 1973.
M. Richards, Welsh Administrative and Territorial Units (Cardiff: University of Wales Press, 1969), p. 117.
LLANFAIRFECHAN Preferred English M. Richards, Welsh Administrative and Territorial Units (Cardiff: University of Wales Press, 1969), p. 117.
CAERNARVONSHIRE Administrative County M. Richards, Welsh Administrative and Territorial Units (Cardiff: University of Wales Press, 1969), p. 117.
LLANFAIRFECHAN AP/CP Parish-level Unit M. Richards, Welsh Administrative and Territorial Units (Cardiff: University of Wales Press, 1969), p. 117.
GB Historical GIS / University of Portsmouth, Llanfairfechan UD through time | Census tables with data for the Local Government District, A Vision of Britain through Time.


Many countries, including Singapore, are presently grappling with the problem of deliberate online falsehoods which has become a serious global issue, said Deputy Speaker of Singapore Parliament Charles Chong today. He was giving his opening remarks as the chairman of the Select Committee on Online Deliberate Falsehoods on the first day of public hearings today.
"It is a complex problem, affecting us in many different ways," he said.
The Select Committee will hear about the different dimensions of the problem over the next three days.
"The committee will hear from witnesses who can help in understanding different aspects of the problem, from media to technology, racial and religious relations to national security," he said.
There are also experts from overseas who will be speaking on the issue. Noteworthy speakers tomorrow include Dr Janis Berzins, director of Center for Security and Strategic Studies at the National Defense Academy of Latvia, Ukraine Crisis Media Center's Nataliia Popovych and Oleksiy Makhuhin and Mr Ben Nimmo, information defense fellow with the Atlantic Council's Digital Forensic Research Lab.
Notably, the Select Committee on Deliberate Falsehoods has attracted the highest number of written representations (164) compared with previous committees. It will also have the highest number of public hearing days (eight) as well as the number of witnesses invited to give oral representations (79 to date) in Singapore's history.
"They represent a cross-section of the representations we received; in some cases, there were representations which required further clarification. We may revise the witness list as the hearings progress," Chong said.
The Committee will examine and report on the phenomenon of using digital technology to deliberately spread falsehoods online, the motivations and reasons for the spreading of such falsehoods, and the parties engaging in such activities and the consequences that the spread of online falsehoods can have on Singapore, including its institutions and democratic processes.
It will also look at how Singapore can prevent and combat online falsehoods.


text:
Korean Olympics may refer to:

 1988 Summer Olympics, held in Seoul, South Korea
 2018 Winter Olympics, held in Pyeongchang, South Korea
 Korea at the Olympics (IOC code: COR), the unified joint united Koreas at the Olympics
 South Korea at the Olympics (IOC code: KOR) the Republic of Korea
 North Korea at the Olympics (IOC code: PRK) the Democratic People's Republic of Korea

See also
 Korean Paralympics (disambiguation)


Stock fotó: Utazás · turista · lány · barátok · Colosseum · Róma · boldog / Travel tourist girl friends by Colosseum, Rome. Happy girlfriends tourists showing victory hand sign gesture in front of Coliseum. Beautiful young happy blonde girl and multiracial Asian woman, Italy.


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text:
Q: Android how to reuse Socket? how to reuse TCP Socket connection(possible setReuseAddress(true), but not work for me) for download multiple files from Server(Apache, HTTPD) in on connection without disconnect(and connect again).
My code below..
public class ClientThread implements Runnable {
        public void run() {
            try {

                Log.d("ClientActivity", "C: Connecting...");
                SocketAddress sockaddr = new InetSocketAddress(serverIpAddress, SERVERPORT);
                socket = new Socket();

                socket.setReuseAddress(true);
                socket.setKeepAlive(true);
                socket.setSoLinger(true, 3000);
                socket.connect(sockaddr);

                if (socket.isConnected()) {
                    DataInputStream input = new DataInputStream(socket.getInputStream());
                    PrintWriter output = new PrintWriter(new OutputStreamWriter(socket.getOutputStream()));

                    output.println("GET /way/images/profile/1231 HTTP/1.1");
                    output.println("Host: 192.168.1.2");
                    output.println("User-Agent: Java");
                    output.println("Accept: */*");
                    output.println("Connection: Keep-Alive");
                    output.println("");
                    output.flush();

                    String line;
                    File file = new File("/sdcard/aaa.png");
                    if (!file.exists()) {
                        file.createNewFile();
                    }
                    FileOutputStream fileOut = new FileOutputStream(file);
                    System.out.println("Getting first file");
                    while ( (line = input.readLine()) != null ) {
                        System.out.println(line);
                        fileOut.write(line.getBytes());
                    }
                    System.out.println("First file finished");
                    fileOut.flush();
                    fileOut.close();

                    output.println("GET /way/images/profile/1231 HTTP/1.1");
                    output.println("Host: 192.168.1.2");
                    output.println("User-Agent: Java");
                    output.println("Accept: */*");
                    output.println("Connection: Keep-Alive");
                    output.println("");
                    output.flush();

                    file = new File("/sdcard/aaa1.png");
                    if (!file.exists()) {
                        file.createNewFile();
                    }
                    fileOut = new FileOutputStream(file);
                    System.out.println("Getting second file");
                    while ( (line = input.readLine()) != null ) {
                        System.out.println(line);
                        fileOut.write(line.getBytes());
                    }
                    System.out.println("Second file finished");

                    fileOut.flush();
                    fileOut.close();
                    input.close();
                    output.close();

                }
                Log.d("ClientActivity", "C: Closed.");
            } catch (Exception e) {
                Log.e("ClientActivity", "C: Error", e);
                connected = false;
            }

            finally {
                try {
                    Log.d("ClientActivity", "Socket: Closed.");
                    socket.close();
                } catch (IOException e) {
                    e.printStackTrace();
                }
            }
        }
    }

Thanks...

A: setReuseAddress(true) prior to binding the socket allows the socket to be bound even though a previous connection is in a timeout state.
documentation of setReuseAddress
I don't see sense in directly using socket for HTTP communication. I use HttpURLConnection. Works without problems in most cases.

redpajama_set_name: RedPajamaStackExchange


Great Spro Product new for 2018 the Expedition rig wallet is available in two sizes small and large. So weather your a day angler are a hoilday angler there is a wallet to match. Both wallet are made to by maintenance free and fully corrosion resistant. Wallet as two mesh pockets and a selection of clear rig sleeves.
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The prospection and development of residential and retrofit projects in Miami, FL. The U.S. business model is similar to the Brazilian one, selecting local partners and operating within the strategic ambit of its projects.
Among the developments in the pre-launch phase are Residential Marina Inn, with 36 units, the precursor of a revitalization process on Ocean Reef Island (Key Largo, FL), and Villa D'Este by Pininfarina, located on Bay Harbor Island, the first residential development created by a renowned Italian design office in Miami. The project envisages the construction of 29 units, all of which with a view of the bay.