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Questionnaire. (PDF 65 kb

Characteristics of benzimidazole compounds, including their metabolites and solubility-improved formulations, in combination with other drugs for the treatment of human cystic and alveolar echinococcosis based on the published literature from 2008 to date.

<p>Characteristics of benzimidazole compounds, including their metabolites and solubility-improved formulations, in combination with other drugs for the treatment of human cystic and alveolar echinococcosis based on the published literature from 2008 to date.</p

Characteristics of natural compounds for the treatment of human cystic and alveolar echinococcosis based on the published literature.

<p>Characteristics of natural compounds for the treatment of human cystic and alveolar echinococcosis based on the published literature.</p

Characteristics of benzimidazole compounds, including their metabolites and solubility-improved formulations, as monotherapy for the treatment of cystic and alveolar echinococcosis based on the published literature from 2008 to date.

<p>Characteristics of benzimidazole compounds, including their metabolites and solubility-improved formulations, as monotherapy for the treatment of cystic and alveolar echinococcosis based on the published literature from 2008 to date.</p

Multiple sequence alignment of DiACT with homologous proteins.

<p>Alignment of the <i>D</i>. <i>immitis</i> ACT sequence (I3WTW3) with the ACT from <i>W</i>. <i>bancrofti</i> (EJD75047), <i>O</i>. <i>volvulus</i> (EJW73626), <i>B</i>. <i>malayi</i> (P48812), <i>L</i>. <i>loa</i> (O01360), <i>Ascaris suum</i> (BAB68543), <i>Homo sapiens</i> (P20287) and <i>S</i>. <i>bovis</i> (AIE44418). The percentage of sequence identity between <i>D</i>. <i>immitis</i> sequence and the others is indicated. The amino acids conserved in all the sequences are labelled with asterisks, and conservative and semiconservative substitutions are respectively labelled with two and one point. Conserved lysine residues are shaded in yellow. The PLG-binding domain (GDEAQSKRGILTLKY) are shaded in grey.</p

Effect of rDiACT and rDiFBAL on the fibrinolytic system activators expression.

<p>Effect of rDiACT and rDiFBAL on the expression of tPA (A and B) and uPA (C and D) in canine vascular endothelial (A and C) and smooth muscle cells (B and D). Protein extracts from lysed rDiACT or rDiFBAL treated or untreated confluent cell cultures were analyzed by Western blot for tPA and uPA. α-tubulin served as a protein control. Data are shown as representative images or means ± SD from three independent experiments. The asterisk (*) designates significant (p<0.05) differences from control cells. (■) Stimulated cells with 1μg/ml of rDiACT. (■) Stimulated cells with 1μg/ml of rDiFBAL. (■) Non-treated control cells.</p

Multiple sequence alignment of DiFBAL with homologous proteins.

<p>Alignment of the <i>D</i>. <i>immitis</i> FBAL sequence (I3WTW4) with the FBAL from <i>O</i>. <i>volvulus</i> (Q9U9R9), <i>L</i>. <i>loa</i> (J0DRR2), <i>B</i>. <i>malayi</i> (A8P3ES), <i>W</i>. <i>bancrofti</i> (J9EVQ4), <i>A</i>. <i>suum</i> (U1M5S0), <i>Haemonchus contortus</i> (R4H2V1) and <i>S</i>. <i>japonicum</i> (C1LB95). The percentage of sequence identity between <i>D</i>. <i>immitis</i> sequence and the others is indicated. The amino acids conserved in all the sequences are labelled with asterisks, and conservative and semiconservative substitutions are respectively labelled with two and one point. Conserved lysine residues are shaded in yellow.</p

MMP-2 and 9 levels assay.

<p>Representative zymography of MMP-2 (solid bars) and MMP-9 (hatched bars) levels in the culture supernatants from canine endothelial (A) and smooth muscle cells (B) untreated or treated with 10 μg/ml of PLG, 1 μg/ml of rDiACT/rDiFBAL, 1 μg/ml of rDiACT/rDiFBAL + 10 μg/ml of PLG or with 1 μg/ml of rDiACT/rDiFBAL + 10 μg/ml of PLG + 50 mM of the εACA. Note the gelatinolytic bands associated with MMP-2 (72 KDa) and MMP-9 (92 KDa) levels, as well as with the MMP-9 activated form (marked with a white asterisk at 82KDa. The results were expressed as percentage of the MMPs levels in the culture supernatant from negative control cells (100%). Data are shown as representative images or means ± SD from three independent experiments. The asterisk (*) designates significant (p<0.05) differences between rDiACT or rDiFBAL treatment and control groups.</p

Immunolocalization of DiACT and DiFBAL in sections from <i>D</i>. <i>immitis</i> adult worms.

<p>Representative images from three independent experiments of parasite sections incubated with phalloidin-Alexa Fluor 488 (in green, specific binding to ACT) plus the negative or the anti-rDiFBAL rabbit sera and an anti-rabbit IgG-Alexa Fluor 594 (in red). Corresponding transmitted light images are also addressed. Magnification 4X.</p

CnAOEC migration by Wound-Healing assay.

<p>Confluent cell cultures were wounded post-treatment and migration distances were measured at 8 hours. The experiment was carried out in canine endothelial cells untreated or treated with 10 μg/ml of PLG, 1 μg/ml of rDiACT/rDiFBAL, 1 μg/ml of rDiACT/rDiFBAL + 10 μg/ml of PLG or with 1 μg/ml of rDiACT/rDiFBAL + 10 μg/ml of PLG + 50 mM of the εACA. The results were expressed as percentage of the migration ability of the negative control cells (100%). Data are shown as representative images or means ± SD from three independent experiments. The asterisk (*) designates significant (p<0.05) differences between rDiACT or rDiFBAL + PLG treatment and control groups.</p