Number of Pgp molecules in a ''resistant'' and a ''susceptible'' isolate before/after exposure to Glucantime.

<p>The number of Pgp 170 molecules was found in higher numbers in the “resistant” compared to the “susceptible” isolate. This number increased, in both isolates, after exposure to Glucantime.</p

Detection of Pgp 170 by Western Blotting in the 10 <i>Leishmania</i> isolates.

<p>The Pgp 170 molecules (130-200 kDa) were evident in the “resistant” (high Rhod-123 efflux) but not in the “susceptible” (low Rhod-123 efflux) isolates. The “resistant” human isolate (H2) presented lower signal compared to the dog “resistant” isolates. For the evaluation of the Pgp expression in each isolate, the C219 monoclonal antibody and exponential phase promastigotes, at a concentration of 10⁷ parasites, were used.</p

Survival of <i>Leishmania</i> promastigotes in different Amphotericin B concentrations, in time.

<p>Survival of the “resistant” dog isolate D5 (a) and of the “susceptible” dog isolate D1 (b), in different drug concentrations for 6 days.</p

Location of Pgp molecules in the body of <i>Leishmania</i> observed under Confocal Microscopy.

<p><i>Leishmania</i> amastigotes (inside THP-1 infected cells) marked with double immunofluorescence: positive dog serum and FITC (green signal) (a); Pgp 170 molecules marked with the C219 monoclonal antibody and the infrared Cy3 anti-mouse IgG secondary antibody (red signal) (b); overlay of image a and b (c). Image c confirms the location of Pgp 170 molecules on the membrane of the parasite body.</p

Heterogeneity of CTC sub-populations according to CellSearch positivity before treatment initiation.

<p>Heterogeneity of CTC sub-populations according to CellSearch positivity before treatment initiation.</p

Kaplan Meier curves for PFS and OS.

<p>(a) Probability of PFS according to the detection of CTC at baseline by the CellSearch, (b) Probability of OS according to the detection of CTC on disease progression by the CellSearch, (c) Probability of OS according to the detection of EMT phenotype expression at baseline and (e) Probability of OS according to the detection of apoptotic phenotype on disease progression.</p

Univariate and multivariate Cox regression analysis.

<p>Univariate and multivariate Cox regression analysis.</p

Evaluation of the detection of Toll-like receptors (TLRs) in cancer development and progression in patients with colorectal cancer

<div><p>Background</p><p>Toll-like receptors (TLRs) play essential role in innate and acquired immunity, are expressed in various cell types, and are associated with altered susceptibility to many diseases, and cancers. The aim of this study was to investigate <i>TLR2</i> (-196 to-174<i>del</i>), <i>TLR4</i> (Asp299Gly and Thr399Ile) and <i>TLR9</i> (T1237C and T1486C) gene polymorphisms at risk of colorectal cancer (CRC) development and progression.</p><p>Methods</p><p>Peripheral blood was obtained from 397 patients with adjuvant (stage II/III, n = 202) and metastatic (n = 195) CRC. Moreover, blood samples from 50 healthy volunteers and 40 patients with adenomatous polyps were also included as control groups. DNA from patients and controls was analyzed using PCR and PCR-RFLP for genotyping functional polymorphism within <i>TLR2</i>, <i>TLR4</i> and <i>TLR9</i> genotypes.</p><p>Results</p><p><i>TLR2–</i>196 to-174<i>del</i>/<i>del</i> genotype was detected in 76.6% of the patients and was significantly higher that the controls groups (<i>p</i><0.001). <i>TLR4</i> Asp299Gly, <i>TLR4</i> Thr399Ile, <i>TLR9 T1237C</i> and <i>T1486C</i> homozygous genotypes were detected in 70.5%, 70.5%, 61.5% and 61.5% of the patients respectively, and were also significantly higher than that in the control groups (<i>p</i><0.001). All polymorphisms detected were also significantly associated with the metastatic disease (<i>p</i><0.001) leading to shorter overall survival (<i>p</i><0.001); whereas, <i>TLR4</i> Asp299Gly and Thr399Ile polymorphisms were significantly associated with <i>KRAS</i> mutations.</p><p>Conclusions</p><p>The detection of higher frequencies of the <i>TLR2</i>, <i>TLR4</i> and/or <i>TLR9</i> polymorphisms in CRC patients compared with the control groups highlight the role of these polymorphism in CRC development and cancer progression.</p></div