Predicted protein sequences of T22D1.2, <i>clx-1</i> and C50F7.5.

<p>Predicted protein sequences of T22D1.2, <i>clx-1</i> and C50F7.5.</p

Venn diagram showing the number of differentially regulated genes per treatment vs. the untreated negative control (DMSO 1%).

<p>H3: OPC enriched fraction “H3”; RE: crude extract (ethanol: water 1:1). 0.02, 0.2 and 2: concentrations of 0.02 mg/mL, 0.2 mg/mL and 2 mg/mL resp., per treatment. Numbers in brackets indicate the number of differentially regulated genes per group.</p

Transcriptome analysis reveals molecular anthelmintic effects of procyanidins in <i>C</i>. <i>elegans</i>

<div><p>Worldwide, more than 1 billion people are affected by infestations with soil-transmitted helminths and also in veterinary medicine helminthiases are a severe threat to livestock due to emerging resistances against the common anthelmintics. Proanthocyanidins have been increasingly investigated for their anthelmintic properties, however, except for an interaction with certain proteins of the nematodes, not much is known about their mode of action. To investigate the anthelmintic activity on a molecular level, a transcriptome analysis was performed in <i>Caenorhabditis elegans</i> after treatment with purified and fully characterized oligomeric procyanidins (OPC). The OPCs had previously been obtained from a hydro-ethanolic (1:1) extract from the leaves of <i>Combretum mucronatum</i>, a plant which is traditionally used in West Africa for the treatment of helminthiasis, therefore, also the crude extract was included in the study. Significant changes in differential gene expression were observed mainly for proteins related to the intestine, many of which were located extracellularly or within cellular membranes. Among the up-regulated genes, several hitherto undescribed orthologues of structural proteins in humans were identified, but also genes that are potentially involved in the worms’ defense against tannins. For example, T22D1.2, an orthologue of human basic salivary proline-rich protein (PRB) 2, and <i>numr-1</i> (nuclear localized metal responsive) were found to be strongly up-regulated. Down-regulated genes were mainly associated with lysosomal activity, glycoside hydrolysis or the worms’ innate immune response. No major differences were found between the groups treated with purified OPCs versus the crude extract. Investigations using GFP reporter gene constructs of T22D1.2 and <i>numr-1</i> corroborated the intestine as the predominant site of the anthelmintic activity. The current findings support previous hypotheses of OPCs interacting with intestinal surface proteins and provide the first insights into the nematode’s response to OPCs on a molecular level as a base for the identification of future drug targets.</p></div

Overview of primers used for qPCR.

<p>Overview of primers used for qPCR.</p

Quantitative RT-PCR.

<p>Relative normalized expression of <i>ugt-44</i>, <i>pud-1</i>.<i>1</i>, F13E9.4 and T22D1.2 mRNA following the treatment with OPC enriched fraction “H3” or <i>C</i>. <i>mucronatum</i> extract (ethanol: water 1:1; “RE”); 0.02, 0.2 and 2: concentrations of 0.02 mg/mL, 0.2 mg/mL and 2 mg/mL respectively. Data are presented on a log2 scale, the line at 1.0 indicates the expression level of the negative control (DMSO 1%).</p

In Vivo Consumption of Cranberry Exerts ex Vivo Antiadhesive Activity against <i>FimH</i>-Dominated Uropathogenic Escherichia coli: A Combined in Vivo, ex Vivo, and in Vitro Study of an Extract from Vaccinium macrocarpon

For investigation of the molecular interaction of cranberry extract with adhesins of uropathogenic Escherichia coli (UPEC), urine from four volunteers consuming standardized cranberry extract (proanthocyanidin content = 1.24%) was analyzed within ex vivo experiments, indicating time-dependent significant inhibition of 40–50% of bacterial adhesion of UPEC strain NU14 to human T24 bladder cells. Under in vitro conditions a dose-dependent increase in bacterial adhesion was observed with proanthocyanidin-enriched cranberry Vaccinium macrocarpon extract (proanthocyanidin content = 21%). Confocal laser scanning microscopy and scanning electron microscopy proved that <i>V.m.</i> extract led to the formation of bacterial clusters on the outer plasma membrane of the host cells without subsequent internalization. This agglomerating activity was not observed when a PAC-depleted extract (<i>V.m.</i> extract^≠PAC) was used, which showed significant inhibition of bacterial adhesion in cases where type 1 fimbriae dominated and mannose-sensitive UPEC strain NU14 was used. <i>V.m.</i> extract^≠PAC had no inhibitory activity against P- and F1C-fimbriae dominated strain 2980. Quantitative gene expression analysis indicated that PAC-containing as well as PAC-depleted cranberry extracts increased the <i>fimH</i> expression in NU14 as part of a feedback mechanism after blocking <i>FimH</i>. For strain 2980 the PAC-containing extract led to up-regulation of P- and F1C-fimbriae, whereas the PAC-depleted extract had no influence on gene expression. <i>V.m.</i> and <i>V.m.</i> extract^≠PAC did not influence biofilm and curli formation in UPEC strains NU14 and 2980. These data lead to the conclusion that also proanthocyanidin-free cranberry extracts exert antiadhesive activity by interaction with mannose-sensitive type 1 fimbriae of UPEC

Representative images of <i>C</i>. <i>elegans</i> expressing pT22D1.2::GFP depending on treatment with hydroethanolic extract from <i>C</i>. <i>mucronatum</i>.

<p>A: negative control (DMSO 1%); B: treatment with 0.2 mg/mL extract; C: treatment with 2 mg/mL extract. Scale bars = 100 μm.</p

Representative images of <i>C</i>. <i>elegans</i> expressing <i>numr-1</i>::GFP depending on treatment with the hydroethanolic extract from <i>C</i>. <i>mucronatum</i>.

<p>A: negative control (DMSO 1%); B: treatment with 0.2 mg/mL extract; C: treatment with 2 mg/mL extract. Scale bars = 100 μm.</p

Functional annotation clusters including up- and down-regulated genes obtained by DAVID.

<p>Functional annotation clusters including up- and down-regulated genes obtained by DAVID.</p

Antiquorum sensing, antibiofilm formation and cytotoxicity activity of commonly used medicinal plants by inhabitants of Borabu sub-county, Nyamira County, Kenya - Fig 5

<p><b>Summarized bioactivity of various concentrations of methanol extracts of <i>E</i>. <i>buchananii</i> about growth (OD</b>_<b>600</b><b>) (a) and quorum sensing (fluorescence/OD</b>_<b>600</b><b>) (b) response of the <i>E</i>. <i>coli</i> Top10.</b> Data are mean values ± SD. Statistical test: One way ANOVA test (<i>n</i> = 3, *p<0.01, **p<0.05, and ***p<0.001).</p