Issues and Prospects of the Library in Japan : From the Answer of the Examination in Asistant Librarian Training

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<i> In vivo</i> bioluminescence imaging of <i>Leishmania</i> in the C57BL/6 ear model.

<p>Simultaneous follow up of parasite load in the ear dermis and of lesion onset, features and cure in mice inoculated with luciferase-transgenic <i>Leishmania major</i>. 10⁴ luciferase-expressing NIH 173 metacyclic promastigotes were inoculated into the dermis of the right ear of C57BL/6 mice (day 0) and followed for more than 80 days. The bioluminescent signal is displayed as a pseudo-colour image representing light intensities over the body surface area. Red represents the most intense signal while blue corresponds to the weakest one. A, B: Individual follow up of a representative mouse left without any ointment application. Clinical features (A) were simultaneously monitored with parasite load fluctuations assessed by the bioluminescence of luciferase-expressing parasites (B). (C) Bioluminescence quantification of the parasite load (left panel; photons/sec/ear; grey area = background measurement) and “lesion” area (right panel; mm²) were followed for 7 mice left without any ointment and depicted as medians +/−sd. Note the detection of a bioluminescence signal before any significant clinically detectable features. Of note, the so called “lesion” area measured between day 11 and day 15 was still made up of inflammatory processes free of any leukocyte infiltrates.</p

C57BL/6 mice given an application of WR279396 formulation under an occlusive dressing.

<p>10⁴ luciferase-expressing <i>L. major</i> metacyclic promastigotes were inoculated into the dermis of the right ear of C57BL/6 mice (day 0). (a, b) From day eleven post-<i>L. major</i> inoculation, the topical ointment WR279396 was applied directly to parasite-loaded ears and (c) then covered with an adhesive polyurethane dressing (arrow). (d,e) Two independent leaflets of surgical tape were applied directly on the occlusive dressing. This surgical tape permitted maintenance of the dressing and kept the formulation in contact with the parasite-loaded site for two days.</p

Determination of the most suitable regimen for WR279396 application under an occlusive dressing.

<p>Eleven days post-inoculation of 10⁴ luciferase-transfected <i>L. major</i> parasites, three groups of 7 mice were constituted: In the control group, mouse ears were left without any ointment (blue plain line), and in the other group WR279396 was applied on the mouse ear under an occlusive dressing with different application frequencies; either 5 applications/10 days (brown plain line) or 5 applications/5 days (brown dotted line). Bioluminescence evaluation of parasite load was performed over the course of 64 days post-inoculation (A). Note that the rebound of the parasite load in the every day-application group can be associated to a more severe lesion as illustrated by a representative mouse at day 36 post-inoculation (B/5 days). White squares delimit the “lesion” area. (C, D) Parasites loads were analysed with ANOVA whose two factors were the treatment (WR279396 10 days or 5 days) and the period of observation. Pair wise comparisons using t-tests were realized for each combination of factors.</p

Individual follow up of bioluminescence (parasite load) and “lesion” area in mice treated with WR279396 without any dressing.

<p>10⁴ luciferase-expressing <i>L. major</i> metacyclic promastigotes were inoculated into the dermis of the right ear of C57BL/6 mice (n = 10; day 0) treated with WR279396 (5 applications (↑) for 10 days) at day 11 post-inoculation. The parasite load (photons/sec/ear) in individual mice (A, B) and the area (mm2; C) of the lesion displayed by the same mice were followed for 3 months. (A, B, C) Green colour assesses the profile in mice that controlled their ear parasite load, i.e. exhibiting a bioluminescence value<1×10⁶ photons/sec/ear at day 33 (green points in panel A and green lines in panels B and C). In contrast, red colour corresponds to mouse ears that display a high bioluminescence value at day 33 (>1×10⁶ photons/sec/ear; red points in panel A and red lines in panels B and C). Note that lesion area values (C) did not assess any clinical failure except in 1 mouse (arrow). Values obtained for control mice are shown in the grey areas indicated in each graph.</p

Phylogenetic analysis of the bat gammaretrovirus-related sequence.

<p>(A) Schematic representation of the partial genome structure encompassing the pol (almost 3,580 nt encoding, the polymerase of almost 1,190 aa) gene of the porcine endogenous retrovirus (GenBank number Y17013), with black bars corresponding to the longest contig sequences (>900 nt) of bat gammaretrovirus (named Sers gammaretrovirus) identified in samples from b7 (<i>Eptesicus serotinus)</i>. The genomic region amplified by PCR is represented by a dashed bar, and the sequence used for phylogenetic analysis is indicated with an asterisk. (B) Phylogenetic tree produced from the amino-acid alignment based on a selected region (155 aa) of the translated sequence obtained from the PCR product (almost 288 aa, approximate positions 173 to 423 of the pol protein of the porcine endogenous retrovirus). The bat gammaretrovirus-related sequence is indicated in bold, with circles in black indicating bat gammaretroviruses. The scale bar indicates branch length, and bootstrap values ≥70% are shown next to the relevant nodes. The tree is midpoint-rooted for purposes of clarity only. REV, reticuloendotheliosis virus; FeLV, feline leukemia virus; GALV, gibbon ape leukemia virus; F-MuLV, Friend MuLV; R-MuLV, Rauscher murine leukemia virus; M-MuLV, Moloney MuLV; M-CRV, murine type C retrovirus; PreXMRV-1/2, pre-xenotropic MuLV-related virus 1 and 2; PERV-A, porcine endogenous type C retrovirus class A; PERV-B, porcine endogenous retrovirus B; PERV-C, porcine endogenous retrovirus C; RD114, feline RD114 retrovirus; MDEV, <i>Mus dunni</i> endogenous virus; KoRV, koala retrovirus; OOEV, <i>Orcinus orca</i> endogenous retrovirus; BaEV, baboon endogenous virus; RpuRV, <i>Rhinolophus pusillus</i>; RmRV, <i>Rhinolophus megaphyllus</i>; RaRV, <i>Rhinolophus affinis</i> retrovirus; MrRV, <i>Myotis ricketti</i> retrovirus; PaRV, <i>Pteropus alecto</i> retrovirus and MlRV, <i>Megaderma lyra</i> retrovirus.</p

Phylogenetic analysis of VP1 bat rotavirus-related sequences.

<p>(A) Schematic representation of the VP1 segment (almost 3,300 nt encoding the RNA-dependent RNA polymerase of almost 1,090 aa) of the genome of the lamb rotavirus strain Lamb-NT (GenBank number FJ031024), with black bars corresponding to the longest contig sequences (>300 nt) of the bat rotavirus (named Maule rotavirus) identified in specimen b8 (<i>Myotis mystacinus</i>). The genomic region amplified by PCR is represented by a dashed bar, and the sequence used for phylogenetic analysis is indicated with an asterisk. B) Phylogenetic tree produced from the amino-acid alignment based on the partial VP1 sequence (119 aa, positions 964 to 1082 of the VP1 protein of lamb rotavirus strain Lamb-NT) translated from one of the longest HSPs. The bat rotavirus-related sequence is indicated in bold within the various rotavirus groups. The scale bar indicates branch length, and bootstrap values ≥70% are shown next to the relevant nodes. The tree is midpoint-rooted for purposes of clarity only.</p

Identification and distribution of sequences of vertebrate viruses of interest among the various bat specimens and tissue samples analyzed.

<p><b>Legend :</b></p>a<p>Br = brain, Li = liver, Lu = lungs.</p>b<p>Matching (HTS) or targeting (PCR/Sanger) viral genes (in brackets), with S = small, M = medium, L = large or polymerase, RdRp = RNA-dependent RNA polymerase.</p>c<p>+ = positive, - = negative.</p>d<p>PCR products larger than expected.</p>e<p>ND = not done.</p

Distribution of unassembled read sequences after BLASTn analysis.

<p>(A) Percentage of sequences related to the main categories of existing viruses: vertebrate (blue), plant/fungal (green), invertebrate (brown), protozoan (yellow) viruses and bacteriophages (gray), and unassigned viral sequences (no taxonomic data concerning the family available, indicated in red). The total number of viral read sequences is indicated below each pie chart. (B) The percentage of sequences related to the most abundant viral families, indicated in the same colors for each main viral category as in (A): blue = vertebrate, brown = invertebrate, gray = phage. Viral families accounting for less than 2% of total sequences were pooled and represented as the “other” category (in purple), and read sequences with no available data concerning the taxonomic family were considered to be unassigned (in red).</p

Distribution of contig sequences after BLASTx analysis.

<p>(A) Percentage of sequences related to the main categories of existing viruses: vertebrate (blue), plant/fungal (green), invertebrate (brown), protozoan (yellow) viruses and bacteriophages (gray), and unassigned viral sequences (no data available concerning the taxonomic family, indicated in red). The total number of viral contigs is indicated below each pie chart. (B) Percentage of sequences related to the most abundant viral families, indicated by the same color range for each of the main viral categories as in (A) : blue = vertebrate, green = plant/fungal, brown = invertebrate, yellow = protozoan, gray = phage. Viral families accounting for less than 2% of all sequences are pooled in the “other” category (in purple), and read sequences with no available data regarding the taxonomic family are considered to be unassigned (in red).</p