Reactivity of the mAb C6 with FN recombinant fragments.

<p><b>A)</b> Different FN recombinant fragments tested with the mAb C6. <b>B)</b> Reactivity in ELISA of various concentrations of the mAb C6 with the FN recombinant fragment containing the type III repeats B and 8; it fragment B-8 with the mutation Glu1329Ala and fragment B-8 with the mutation Asp1385Glu.</p

Interaction between C6 and the FN recombinant fragment formed by the type III repeats B and 8.

<p><b>A)</b> Schematic representation of the main interactions between the two proteins. The type III domains B and 8 are drawn as green ribbons, the scFv C6 is orange. Residues involved in the binding are reported as sticks, hydrogen bonds between amino acids are shown as dotted black lines. <b>B)</b> Residues of the scFv and of the type III repeats B and 8 that interact. The distances are displayed. <b>C)</b> Representation of the interaction between the type III domains B-8 (green surface) and the scFv C6 (orange surface).</p

Immunohistochemistry experiments using the mAb C6 on cryostat sections.

<p>A) Normal human lung; B) human lung adenocarcinoma; C) Normal human brain; D) Human mesothelioma; E) Human glioblastoma; F) Murine teratocarcinoma. Immunohistochemistry experiments using the recombinant antibody CGS1 on cryostat sections of human glioblastoma (G) and murine teratocarcinoma (H). CGS1 reacts directly with the ED-B (10) and thus, contrary of C6, it also reacts with murine tumours. C6 reacts, in an identical manner of CGS1, with normal and cancer human tissues but not with murine tumours (Modified from ref.[<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0148103#pone.0148103.ref017" target="_blank">17</a>] under CC-BY license, with permission f from John Wiley and Sons, original copyright 2009.). </p

Structures of FN and of the scFv C6.

<p><b>A)</b> Model of the domain structure of a FN subunit. The three different types of repeats, the three sites of alternative splicing (ED-A, ED-B, IIICS) and the specificity of the mAb C6 for the interface between type III repeats B and 8 are shown. <b>B)</b> Sequence of the scFv C6; the linker between the VL and the VH is in red; the CDRs are framed. Amino acids involved in the interaction with C6 are in bold. <b>C)</b> Sequence of the FN type III repeats numbers 7 (blue), B (black) and 8 (red); the various loops between the beta sheets structures are framed. The amino acids involved in the interaction with C6 are in bold. Sequence from <a href="http://www.ncbi.nlm.gov.nuccore/47132556" target="_blank">http://www.ncbi.nlm.gov.nuccore/47132556</a>.</p

ABCB1 Structural Models, Molecular Docking, and Synthesis of New Oxadiazolothiazin-3-one Inhibitors

Docking methods are powerful tools for in silico screening and drug lead generation and optimization. Here, we describe the synthesis of new inhibitors of ABCB1 whose design was based on construction and preliminary confirmation of a model for this membrane transporter of the ATP-binding cassette family. We chose the strategy to build our three-dimensional model of the ABCB1 transporter by homology. Atomic coordinates were then assayed for their reliability using the measured activity of some oxadiazolothiazin-3-one compounds. Once established their performance by docking analysis, we synthesized new compounds whose forecasted activity was tested by MTT and cytofluorimetric assays. Our docking model of MDR1, MONBD1, seems to reliably satisfy our need to design and forecast, on the basis of their LTCC blockers ability, the inhibitory activity of new molecules on the ABCB1 transporter

Numbers (mean ± standard error) of apoptotic (annexin V positive) and necrotic (propidium iodide positive, annexin V negative) human peripheral blood leukocytes per 100 gated cells after <i>in vitro</i> incubation with FLIP7.

<p>Numbers (mean ± standard error) of apoptotic (annexin V positive) and necrotic (propidium iodide positive, annexin V negative) human peripheral blood leukocytes per 100 gated cells after <i>in vitro</i> incubation with FLIP7.</p

Sequences of <i>C</i>. <i>vicina</i> AMPs determined by transcriptome analysis and peptide sequencing.

<p>Sequences of <i>C</i>. <i>vicina</i> AMPs determined by transcriptome analysis and peptide sequencing.</p

Mass spectrometric characteristics and activity profiles of FLIP7 anti-biofilm AMPs.

<p>Mass spectrometric characteristics and activity profiles of FLIP7 anti-biofilm AMPs.</p

Cell killing (TTC assay) and biofilm eradicating (crystal violet assay) activity of FLIP7 fractions against <i>E</i>. <i>coli</i> and <i>S</i>. <i>aureus</i> biofilms.

<p>Cell killing (TTC assay) and biofilm eradicating (crystal violet assay) activity of FLIP7 fractions against <i>E</i>. <i>coli</i> and <i>S</i>. <i>aureus</i> biofilms.</p

Photomicrographs of biofilms formed by three bacterial strains in normal conditions and in the presence of FLIP7.

<p><i>A</i>. <i>baumannii</i> 28, <i>E</i>. <i>coli</i> ATCC 25922 and <i>S</i>. <i>aureus</i> 203 biofilms grown as described in Materials and Methods section were incubated 24 hours in the culture medium (control) or the medium supplemented with 4 mg/mL of FLIP7 and photographed using Nomarski optics at 400-fold and 1000-fold (inset) magnification. All three strains formed dense biofilms containing live bacteria attached to the glass surface (control). At the same time, FLIP7 presence in the medium led to the destruction of the biofilm, in the remains of which are visible mainly cells that have lost their characteristic shape.</p