Additional file 3: of The mitochondrial genome of Globodera ellingtonae is composed of two circles with segregated gene content and differential copy numbers

Alignment of the start of the shared sequence region between Globodera ellingtonae mtDNA-I and mtDNA-II. (PDF 105 kb

Extreme precipitation in low mountain ranges in Central Europe: a comparative study between the Vosges and the Ore mountains

of the doctoral dissertation Extreme precipitation is related to flooding which is one of the most frequent natural hazards in Central Europe. Detailed understanding of extreme precipitation is the precondition for an efficient risk management and more precise projections of precipitation, which include uncertainties, especially at regional scale. The thesis focuses on extreme precipitation in the Ore Mountains (OM) and the Vosges Mountains (VG); two low mountain ranges in Central Europe experiencing orographic effect on precipitation. Based on state of the art about precipitation in OM and VG, a currently missing analysis of the temporal distribution of precipitation in VG was needed prior to the analysis of extremes. The original dataset of daily precipitation totals from 14 weather stations used in the initial study was extended to 168 stations covering a broader area of VG. The study of temporal distribution of precipitation during 1960-2013 led to a classification of stations: (i) mountainous stations with winter maxima and highest mean annual totals due to orographic enhancement of precipitation, (ii) stations on leeward slopes with two maxima (summer and winter), (iii) lee side stations with summer maxima and lowest mean annual totals due to rain shadow and more continental character, and..

Gene expression in pathways altered only by <i>T. b. rhodesiense</i> inhibited for brucipain activity.

<p>These gene sets are canonical representations of a biological process compiled by domain experts. PAGE gene set analysis was performed using the DIANE 6.0 software using the PAGE algorithm. The archive of gene sets used with this algorithm for this analysis is from the MSigDB. Significance of functions and pathways was calculated using the right-tailed Fisher's Exact Test. The p-value was calculated by comparing the number of user-specified genes of interest participating in a given function or pathway relative to the total number of occurrences of these genes in all functional/pathway annotations stored in the knowledge base.</p><p><b>n.s.: not significant</b></p

Real-time TEER changes in HBMEC silenced for PAR-2 gene expression.

<p><i>F2RL1</i> RNAi transfected HBMEC grown in 8W1E ECIS chambers were incubated overnight with <i>T. b. rhodesiense</i> IL1852. The changes in TEER relative to matched HBMEC control containing scrambled siRNA construct (red line) and HBMEC silenced for <i>F2RL1</i> (blue line) in the presence of parasites are shown.</p

PMT from <i>Pasteurella multocida</i> blocks <i>T. b. rhodesiense</i> induced changes in HBMEC TEER.

<p>HBMEC grown in 8W10E+ ECIS chambers were incubated with <i>T. b. rhodesiense</i> IL1852. Shown are the changes in real-time TEER measured by ECIS. A) HBMECs incubated with <i>T. b. rhodesiense</i> IL1852 in the absence (blue line) or continuous presence of PMT (30 ng/ml) (red line). B) HBMECs incubated with <i>T. b. rhodesiense</i> IL1852 was incubated with HBMECs untreated (blue line) or pretreated with PMT (30 ng/ml) (red line). The data are represented as the average change in TEER±EM (n = 3). The changes in TEER are represented as the average change in TEER±SEM (n = 2).</p

Pathway changes altered by <i>T. b. rhodesiense</i> inhibited for brucipain activity.

<p>Significantly regulated, functional pathway clusters were generated from wild-type (WT <i>Tbr</i>) <i>T. b. rhodesiense</i> or parasites K11777 inhibited for brucipain activity (KT <i>Tbr</i>) gene sets using PAGE gene set analysis. Pathways that were significantly up-regulated (red) or down-regulated (green) are shown. Cluster group numbers are shown on the left, while the grading scale is in the upper right.</p

Annotation of pathway genes significantly expressed in HBMEC only in response to <i>T. b. rhodesiense</i> inhibited for brucipain activity.

<p>Raw microarray data were subjected to Z normalization and tested for significant changes. Genes were determined to be differentially expressed after calculating the Z ratio and fdr.</p><p><b>n.s.: not significant</b></p

<i>T. b. rhodesiense</i> transmigration across HBMEC silenced for <i>F2RL1</i> expression by RNAi.

<p>Using laser capture microdissection 15 individual HBMEC expressing GFP in the <i>PAR2</i>-silenced and control siRNA cultures were collected and the RNA extracted. (A) Based on qRT-PCR using pre-designed <i>F2RL1</i> primers and normalized to <i>ACTB</i> (β-actin transcripts), PAR-2 gene expression was reduced by over 95%. (B) <i>T. b. rhodesiense</i> IL1852 was incubated for 16h in triplicate with HBMEC monolayers silenced for PAR-2 expression (<i>F2RL1</i> siRNA), with a matched scrambled siRNA control (control siRNA) construct, or with untreated HBMEC (no Rx control) and examined parasite transmigration.</p

Proposed model for African trypanosome-induced BBB dysfunction.

<p>We hypothesize that parasite proteases trigger GPCRs (i.e. PARs?) via Gαq activation, which leads to PLC-mediated Ca²⁺ release from intracellular stores. The increase in intracellular calcium leads to calmodulin (CaM) activation of myosin light chain kinase (MLCK), ultimately leading to cytoskeletal changes and barrier dysfunction. Ca²⁺-independent activation of the cytoskeleton mediated by Ras-superfamily GTPases (i.e. RhoA) is also possible via p63RhoGEF. Parasite and/or host-derived proteases may also contribute by degrading or altering adherens junction (AJ) and tight junction (TJ) proteins.</p

Functional overview of significant categorized MSigDB gene sets based on the pathways altered by both wild-type and <i>T. b. rhodesiense</i> inhibited for brucipain activity.

<p>A functional overview of the MSigDB gene sets categorized into a small number of selected gene families whose members a common feature such as homology or biochemical activity. They do not necessarily have common origins. Annotation of pathway genes significantly expressed in HBMEC only in response to <i>T. b. rhodesiense</i> inhibited for brucipain activity are shown in <b>bold font.</b></p