Accession Numbers

<p>ND, not determined.</p

Properties of the Neuraminidase of H1N1 Viruses from the 2007–2008 Season

<p><i>p-</i>Values are the result of the Student's t test between the mean of K_m, V_m, or K_i of the sensitive viruses versus either the resistant ones or the pre-2007–2008 viruses. Student's t test was also performed between the mean K_m of the sensitive viruses versus the mean K_m of the early seasonal virus #0006/07.</p><p>Amino acids that differ from the NC99 strain are shown in bold.</p>a<p>Except for the reference vaccine strains A/New Caledonia/20/99(H1N1) (NC99) and A/Solomon Islands//2006(H1N1) (SI06), all other viruses were A(H1N1) clinical isolates from the NIC (Northern-France) named by order number and year of isolation. Virus 0692/07 is an isolate from the 2006–2007 season.</p>b<p>Week of sampling of specimen during the 2007–2008 season.</p>c<p>IC50 determined essentially as described in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1000103#ppat.1000103-RameixWelti1" target="_blank">[20]</a>,<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1000103#ppat.1000103-Potier1" target="_blank">[30]</a> using the MUNANA substrate at a final concentration of 100 μM.</p>d<p>The Michaelis-Menten constant K_m was determined by enzymatic kinetic analyses performed on inactivated virus suspensions. Briefly, kinetics were performed using MUNANA concentrations ranging from 5 to 100 μM. Initial velocity of the reaction was calculated and plotted as a function of the MUNANA concentration. K_m and V_m were calculated using a nonlinear regression of the curve according to the Michaelis-Menten equation. V_m values were expressed in arbitrary units (U/sec).</p>e<p>Hemagglutination (HA) titers were determined in duplicate by standard procedures using guinea pig red blood cells. Titers are expressed as the mean of the reciprocal of the last virus dilutions showing hemagglutination.</p>f<p>K_i determinations rely on enzymatic kinetic analyses. The kinetics were performed in the presence of variable concentrations of inhibitor (0 to 2,000 nM) and a constant MUNANA concentration (20 μM). Calculation of the K_i was performed by nonlinear regression of the plot of the initial velocity as a function of the concentration of inhibitor.</p>g<p>K_m and V_m values are given as the mean of 2, 3^*, 4^**, or 7^*** determinations.</p>h<p>Values correspond to the mean ± standard deviation of the mean K_m or V_m values of the 5 pre-2007–2008 viruses , the 6 sensitive viruses, and the 11 resistant viruses. Means and <i>p</i>-values apply to virus groups indicated by —.</p>i<p>K_i values were determined once except for the SI06 reference strain, for which three independent determinations were performed, and the 0497/07 and 644/07 viruses, for which two independent determinations were performed.</p>j<p>Values correspond to the mean ± standard deviation of the mean K_i values of the 5 pre-2007–2008 viruses, the 6 sensitive viruses, and the 11 resistant viruses. Means and <i>p</i>-values apply to virus groups indicated by —.</p><p>ND, not determined; NS not significant.</p

Growth Kinetics of H1N1 Viruses from the 2007–2008 Season Sensitive or Resistant to Oseltamivir.

<p>Two sensitive (A/Paris/497/2007 and A/Paris/1149/2008) and two resistant (A/Paris/644/2007 and A/Paris/1170/2008) viruses from the 2007–2008 influenza season, as well as the reference strain A/Solomon Islands/3/2006 and an isolate from the 2003–2004 season (A/Paris/650/2004), were amplified and titrated on MDCK cells. The indicated viruses were then used to infect MDCK SIAT-1 cells <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1000103#ppat.1000103-Matrosovich1" target="_blank">[31]</a> at an m.o.i. of 0.001 and incubated for 72 hours at 35°C in the presence of 1 μg/ml TPCK trypsin. At the indicated time points, the supernatants were harvested and virus titers were determined by plaque assays on MDCK cells.</p

Phylogenetic tree generated with Neighbor-joining (NJ) method on a 522-bp fragment of the HTLV-1 gp-21 <i>env</i> gene for 25 HTLV-1 available sequences including the 2 sequences generated in this work (NCP201 and EM5).

<p>HTLV-1 strains were aligned with DAMBE software (version 4.2.13). The final alignment was submitted to the Model test program (version 3.6) to select, according to the Akaike Information Criterion (AIC), the best model to apply to phylogenetic analyses. The selected model was the Tamura Nei. Bootstrap values were calculated for 1,000 replicates and indicate frequencies of occurrence for 100 trees (bootstrap ≥50%). The branch lengths are drawn to scale with bar indicating 0.01-nucleotide replacement per site. The ATK-1 strain was used as outgroup. The NCP201 and EM5 strains belong to the “Solomon/Vanuatu/New Caledonian” sub-clade, while the two other “Australian” and “Papua New Guinean” clades exist within HTLV-1c subtype. (Genbank accession nos. KX905202 and KX905203).</p

Human T-cell lymphotropic virus type 1 serologic confirmation by Western blot.

<p>HTLV-1 seroreactivity patterns obtained by Western blot with recombinant GD21 (common to HTLV-1 and HTLV-2) and two synthetic peptides specific for HTLV-1 (MTA-1) and HTLV-2 (K55). Lane 1, HTLV-1 positive control; lane 2, HTLV-2 positive control; lane 3, HTLV-1/2 negative control; lanes 4–6, plasma samples from the HTLV-1 positive women from New Caledonia (NCP91, NCP173 and NCP201) displaying a strong reactivity to GD21 and to p19, p24, p26, p28, p32, p36 plus rgp46-I (MTA-1).</p

Illustration of a Border Collie pedigree segregating PRA constructed by the Cyrillic 2

1 software. This pedigree is constituted of 80 dogs, 33 dogs are affected (30 males and 3 females).<p><b>Copyright information:</b></p><p>Taken from "Progressive Retinal Atrophy in the Border Collie: A new XLPRA"</p><p>http://www.biomedcentral.com/1746-6148/4/10</p><p>BMC Veterinary Research 2008;4():10-10.</p><p>Published online 3 Mar 2008</p><p>PMCID:PMC2324077.</p><p></p

Map of the western Pacific region.

<p>The Australo-Melanesia region comprises the islands group extending from Papua New Guinea to New Caledonia including Solomon Islands and Vanuatu archipelago plus Australia, where HTLV-1 epidemiological and clinical situation has been investigated.</p

Human T-cell lymphotropic virus type 1 serologic screening tests.

<p>Human T-cell lymphotropic virus type 1 serologic screening tests.</p

Phylogenetic tree generated with neighbor-joining (NJ) method on a 2,346-bp fragment of the HTLV-1 <i>gag-tax</i> concatenated genes for 29 HTLV-1 available sequences including the 2 sequences generated in this work (NCP201 and EM5).

<p>HTLV-1 strains were aligned with DAMBE software (version 4.2.13). The final alignment was submitted to the Model test program (version 3.6) to select, according to the Akaike Information Criterion (AIC), the best model to apply to phylogenetic analyses. The selected model was the GTR. The numbers at some nodes of the tree (bootstrap values) were calculated for 1,000 replicates and indicate frequencies of occurrence for 100 trees (bootstrap ≥50%). The branch lengths are drawn to scale with bar indicating 0.01-nucleotide replacement per site. The ATK-1 strain was used as outgroup. The NCP201 and EM5 strains belong to the Australo-Melanesian HTLV-1c subtype and clustered with the strains previously characterized in Vanuatu (ESW44) the Solomon Islands (Mel5). Strains from Central Australia constitute a second clade (Genbank accession nos. KX905202 and KX905203).</p

Virus budding from infected myotubes and myoblasts.

<div><p><b>A to J</b>. Transmission electron microscopy (TEM) was performed on non-infected myotubes (<b>A</b>, <b>B</b>), on myotubes infected for 20h with the A/California/7/2009 (<b>C</b>, <b>D</b>, <b>E</b>, <b>F</b>) or the Paris/1149/2008 (<b>G</b>, <b>H</b>) isolates, and on MDCK-SIAT cells, 16h after infection with the A/California/7/2009 isolate (<b>I</b>, <b>J</b>). Arrowheads: budding virions or virions in close contact with the cell. Arrows: dense nuclear aggregates. Stars: nucleoli. Bars: A to E and G to J: 2µm; F: 100nm.</p> <p><b>K to T</b>. Correlative light and electron microscopy (CLEM) was carried out on non-infected myoblasts (<b>K</b>, <b>L</b>) and on myoblasts infected for 20h with the A/California/7/2009 (<b>M</b>, <b>N</b>, <b>O</b>, <b>P</b>) or the A/Paris/1149/2008 (<b>Q</b>, <b>R</b>, <b>S</b>, <b>T</b>) isolates. <b>K</b>, <b>M</b>, <b>Q</b>. Immunofluorescence imaging of myoblasts expressing desmin (red) and/or NP (green). <b>L</b>. Transmission electron microscopy imaging of a non-infected myoblast observed in K showing an intact nucleus. <b>N</b>, <b>O</b>, <b>P</b> and <b>R</b>, <b>S</b>, <b>T</b>. Transmission electron microscopy imaging of the cells observed in M and Q, respectively, showing budding virions or nuclear aggregates. Arrowheads: budding virions or virions in close contact with the cell. Arrows: dense nuclear aggregates. Stars: nucleoli. Bars: K, M, Q: 50µm; N, R: 20 µm; L, O, P, S: 2µm; T: 200nm. </p></div