Radiotherapy for Ledderhose disease:Results of the LedRad-study, a prospective multicentre randomised double-blind phase 3 trial

Background and purpose: Radiotherapy is considered a treatment option for Ledderhose disease. However, its benefits have never been confirmed in a randomised controlled trial. Therefore, the LedRad-study was conducted. Materials and methods: The LedRad-study is a prospective multicentre randomised double-blind phase three trial. Patients were randomised to sham-radiotherapy (placebo) or radiotherapy. The primary endpoint was pain reduction at 12 months after treatment, measured with the Numeric Rating Scale (NRS). Secondary endpoints were pain reduction at 6 and 18 months after treatment, quality of life (QoL), walking abilities and toxicity.Results: A total of 84 patients were enrolled. At 12 and 18 months, patients in the radiotherapy group had a lower mean pain score compared to patients in the sham-radiotherapy group (2.5 versus 3.6 (p = 0.03) and 2.1 versus 3.4 (p = 0.008), respectively). Pain relief at 12 months was 74% in the radiotherapy group and 56% in the sham-radiotherapy group (p = 0.002). Multilevel testing for QoL scores showed higher QoL scores in the radiotherapy group compared to the sham-radiotherapy group (p &lt; 0.001). Moreover, patients in the radiotherapy group had a higher mean walking speed and step rate with barefoot speed walking (p = 0.02). Erythema, skin dryness, burning sensations and increased pain were the most frequently reported side effects. These side effects were generally graded as mild (95%) and the majority (87%) were resolved at 18 months follow-up.Conclusion: Radiotherapy for symptomatic Ledderhose disease is an effective treatment resulting in a significant pain reduction, improvement of QoL scores and bare feet walking abilities, in comparison to sham-radiotherapy.</p

In Vivo Monitoring of mRNA Movement in Drosophila Body Wall Muscle Cells Reveals the Presence of Myofiber Domains

Background: In skeletal muscle each muscle cell, commonly called myofiber, is actually a large syncytium containing numerous nuclei. Experiments in fixed myofibers show that mRNAs remain localized around the nuclei in which they are produced. Methodology/Principal Findings: In this study we generated transgenic flies that allowed us to investigate the movement of mRNAs in body wall myofibers of living Drosophila embryos. We determined the dynamic properties of GFP-tagged mRNAs using in vivo confocal imaging and photobleaching techniques and found that the GFP-tagged mRNAs are not free to move throughout myofibers. The restricted movement indicated that body wall myofibers consist of three domains. The exchange of mRNAs between the domains is relatively slow, but the GFP-tagged mRNAs move rapidly within these domains. One domain is located at the centre of the cell and is surrounded by nuclei while the other two domains are located at either end of the fiber. To move between these domains mRNAs have to travel past centrally located nuclei. Conclusions/Significance: These data suggest that the domains made visible in our experiments result from prolonged interactions with as yet undefined structures close to the nuclei that prevent GFP-tagged mRNAs from rapidly moving between the domains. This could be of significant importance for the treatment of myopathies using regenerative cellbase

Simultaneous expression (and export) of two proteins of the same family in a single blood stage parasite (trophozoites or schizonts) of double-gene tagging (DGT) mutants.

<p>These mutants contain the following pairs of genes tagged with either mCherry or GFP: <i>fam-a1/fam-a2</i> (2 independent mutants; panel A, B), <i>fam-b1/fam-b2</i> (panel C<i>) and pir1/pir3</i> (panel D); RMgm ID as indicated in <b><a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1005917#ppat.1005917.t002" target="_blank">Table 2</a></b>. Localisation of mCherry-tagged members in the trophozoite stage (upper two rows) and in the schizont (low row) stage. Parasite nuclei are stained with Hoechst, BF bright field. scale bar: 5μm.</p

Expression of proteins encoding blood-stage exported proteins in blood- and liver-stages

<p>Expression of proteins encoding blood-stage exported proteins in blood- and liver-stages</p

Expression of fluorescently-tagged proteins of multigene families in liver stages of single-gene tagging (SGT) mutants at 48hpi in cultured hepatocytes (Huh7).

<p><b>A</b>. Fluorescence-microscopy analysis of members of the <i>fam-a</i>, <i>fam-b</i> and <i>pir</i> multigene family in live liver-stages. The parasites expressing mCherry-tagged Fam-b2 and PIR1 also express cytoplasmic GFP (cyt GFP; green). <b>B</b>. IFA-analysis of fixed liver-stages using anti-mCherry (red) anti-PbEXP1 (green) antibodies. PbEXP1 is a parasitophorous vacuole membrane resident protein. <b>C</b> Fluorescence-microscopy analysis of expression of SMAC and IBIS, exported proteins encoded by single-copy genes in live liver-stages. Nuclei are stained with Hoechst-33342 (blue); scale bar: 10μm.</p

Maximum likelihood phylogeny of <i>fam-a</i> gene sequences from <i>Plasmodium</i> spp.

<p>The tree was estimated using RAxML and a GTR+Γ model. Branches subtended by nodes with >75 bootstrap support are shown in bold. Robust basal nodes are indicated by black squares with bootstrap proportions (above node) and Bayesian posterior probabilities (beneath node). At right, coloured blocks indicate the species to which a terminal node belongs. Clades of orthologs that display positional conservation are indicated with green blocks; where a sequence has been lost secondarily in a species is shown by an ‘X’. The tree is rooted using an out-group comprising single copy <i>fam-a</i> orthologs from primate <i>Plasmodium</i> species. The phylogeny is subdivided into four sections: genes located at the conserved, ‘ancestral’ locus on chromosome 13 (below line <i>i</i>); genes found at loci conserved across RMP species (between lines <i>i</i> and <i>ii</i>); and a robust clade of species-specific paralogs derived from a conserved locus on chromosome 6 or 13 (between lines <i>ii</i> and <i>iii</i>); a robust clade of species-specific paralogs derived from a conserved locus on chromosome 8 (above line <i>iii</i>).Transcription levels (shown as different coloured and sized circles) in blood stages are shown for individual genes based on RNAseq data (FPKM values) (from [<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1005917#ppat.1005917.ref033" target="_blank">33</a>] and <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1005917#ppat.1005917.s007" target="_blank"><b>S1</b> Table</a>). Expression levels as shown by four different sized circles: Class 1 (smallest circle): 2-8x the threshold level; class 2: 8-16x the threshold; class 3 (largest circle): >16x the threshold.</p

Transcription of <i>pir</i>, <i>fam-a</i> and <i>fam-b</i> genes based on RNAseq data of different blood stages of two <i>P</i>. <i>berghei</i> ANKA reference lines.

<p><b>A</b>. Features of transcription of <i>pir</i>, <i>fam-a</i> and <i>fam-b</i> genes in the two <i>P</i>. <i>berghei</i> reference lines (line 1 and line 2) based on RNAseq data (from[<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1005917#ppat.1005917.ref033" target="_blank">33</a>] and shown in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1005917#ppat.1005917.s007" target="_blank">S1 Table</a>). Transcribed genes are genes with an FPKM value above the cut-off level of 21. Total transcript abundance is the sum of all FPKM values observed in the different blood stages (see <b>B</b>). The fold up-down regulation is based on the difference in FPKM values of individual genes between blood stages of the two different parasite lines (see <b><a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1005917#ppat.1005917.s001" target="_blank">S1 Fig</a></b>). <b>B.</b> Percentage of genes transcribed in the different blood stages (see <b>A</b>). Ring, red; trophozoite, green; schizont, purple; gametocyte, black. <b>C.</b> Total transcript abundance in the different blood stages: mean and standard deviation of total transcript abundance of all FPKM values observed in the different blood stages (see <b>A</b>). <b>D.</b> Percentage of non-transcribed genes (light grey) and genes with less (grey) or more (black) than 1.5x difference in transcript abundance between blood stages of two different parasite lines (see <b>A</b>). The coloured circles show the genes with >1.5 fold down-or upregulation in the four different blood stages (see <b>B</b>).</p