Additional file 1: of Emerging good practices for Translatability Assessment (TA) of Patient-Reported Outcome (PRO) measures

Table S1 Terminology . Table S2 Definitions. Table S3 Steps used by each organization. Table S4 People involved. Table S5 Timing of assessment. Table S6 Review criteria. Table S7 Recommendations (DOCX 56 kb

Expression profile of the nine <i>pmp</i> genes and <i>ompA</i> throughout the development of <i>C. trachomatis.</i>

<p>Reference strain L₂/434 is represented in panel A and E/Bour in panel B. Values represent the mean±SEM based on three independent experiments for time points of 2, 6, 12, 18, 24, 36, and 48 h post infection. See <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000878#s2" target="_blank">methods</a> for details.</p

Predicted <i>pmpF</i> promoter sequence for reference and clinical strains.

<p>Sequences are for reference strains E/Bour and L₂/434, and clinical strains E/537C-05, E/S-141, E/CS-500-96, and L₂. The predicted transcription promoter for <i>pmpF</i> is located within a 100% conserved region of the <i>pmpG/pmpF</i> IGR, where putative -10 and -35 elements are in blue characters and boxed. Potential A/T spacer region is underlined, and the predicted transcription start site is shown in a larger font below a red asterisk. The putative RBS for <i>pmpF</i> is in orange characters, and the putative RNase E cleavage sites are highlighted in grey. Numbers represent positions relative to the start codon of <i>pmpF</i> (highlighted in yellow). The start codon of <i>pmpG</i> is highlighted in blue.</p

Distribution/Location of the putative RNase E cleavage sites within the <i>pmpFE</i> operon coding sequence.

<p>The sequence is for reference strains E/Bour and L₂/434 and for clinical strains E/537C-05, E/S-141, E/CS-500-96 and L₂. Black vertical lines represent all RNase E cleavage sites conserved among all strains under study; green vertical lines show the ones only conserved among the four “E” strains; orange vertical lines represent those specific solely for both L₂ strains. Numbers represent nucleotide positions relative to the start codon of <i>pmpF.</i></p

Oligonucleotide primers used for kRT-PCR

a<p>Primers designed based on each <i>pmp</i> sequence of reference strains E/Bour and L₂/434 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000878#pone.0000878-Gomes1" target="_blank">[12]</a>.</p>b<p>Primers designed only for strain L₂.</p>c<p>Primers designed based on the <i>ompA</i> sequence of reference strains E/Bour and L₂/434 (GenBank Accession No. X52557 and M14738, respectively).</p>d<p>Primers designed based on the <i>16SrRNA</i> sequence of reference strains E/Bour and L₂/434 (GenBank Accession No. D85722 and U68443, respectively).</p

Clinical and microbiologic characteristics of female adolescents from whom sera was used for determining the immunoreactivity against rPmpD and rPmpF

a<p>Patients were adolescents 14–19 years of age who had a <i>C. trachomatis</i> infection with only one <i>ompA</i> genotype as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000878#s2" target="_blank">methods</a> or were uninfected;</p>b<p>The diagnosis of cervicitis was based on physical findings consistent with cervicitis as determined by the examining physician; all adolescents infected with <i>C. trachomatis</i> had cervicitis, and none of these patients complained of any symptoms;</p>c<p>A cervical discharge was noted by the examining physician; none of these patients had clinical signs or symptoms consistent with upper genital tract disease.</p

Dot-Blot of serum immunoreactivity against recombinant (r)PmpD and rPmpF.

<p>Sera was obtained from adolescents singly infected and uninfected with a different <i>C. trachomatis</i> clinical strain as described previously <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000878#pone.0000878-Gomes3" target="_blank">[17]</a> (see also <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000878#s2" target="_blank">methods</a>). rPmpD and rPmpF concentrations were standardized for use on the blots. Immunoreactivity to each fusion protein for sera from patients infected with strain Ba (n = 3), D (n = 3), E (n = 8), F (n = 5), G (n = 1), Ia (n = 1), J (n = 2) or K (n = 3) are shown. Of note is that immunoreactivity was consistent for sera from patients infected with the same clinical strain except for strain F (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000878#pone-0000878-t002" target="_blank">Table 2</a>); all eight patients infected with strain E were reactive to rPmpD.</p

Expression profile of <i>pmp</i> and <i>ompA</i> genes throughout the development of <i>C. trachomatis</i> clinical strains.

<p>(A) Strain L₂ shares the same <i>ompA</i> genotype as L₂/434; and strains E/537C-05 (B), E/S-141 (C) and E/CS-500-96 (D) share the same <i>ompA</i> genotype as E/Bour. Values represent the mean±SEM based on three independent experiments for time points of 2, 6, 12, 18, 24, 36, and 48 h post infection. See <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000878#s2" target="_blank">methods</a> for details.</p