Titled extracts of cynara scolymus for use in the treatment of mesothelioma
A technology of extract, artichoke, applied in the field of artichoke titration extract
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[0059] The preparation of such extracts is generally known to those skilled in the art and need not be described in particular detail in this disclosure. For the purposes of the present invention, any of the above-indicated extracts prepared according to conventional techniques may be used.
[0060] In particular, for the purposes of the present invention, the extract may also be replaced by a fraction or a mixture of fractions of the artichoke extract described above or a mixture of an artichoke extract and more than one extract fraction, provided that the above disclosed titration Standard is fine.
[0061] Considering the variability in plant extracts that can arise from climatic conditions, environmental conditions, from differences in cultivation sites and / or cultivation techniques, or even from different varieties of cultivated plants, it is therefore important for clinical use that Standardizing the product and determining standardized parameters enables the provision ...
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[0169] 1. Method
[0170] 1.1. Cell Lysis and Western Blotting.
[0171] Lysis buffer NP40 (50mM Tris-HCl pH 7.4, 150mM NaCl, 1% NP-40, 1mM EGTA, 1mM EDTA) supplemented with protease and phosphatase inhibitors (5mM PMSF, 3mM NaF, 1mM DTT, 1mM NaVO4) ) for 30 min on ice. Equal amounts of total protein extracts (30 μg) were resolved by denaturing electrophoresis on 8% polyacrylamide gels (SDS-PAGE) and transferred to nitrocellulose membranes over 2 hours. Membranes were blocked with 5% milk solution dissolved in TBS-Tween_200.05% for 1 hour and incubated with specific primary antibodies. The following primary antibodies were used: anti-β-actin (A-2228, SIGMA), anti-pSTAT3 (Tyr-705) (sc8059, Santa Cruz) and anti-STAT3 (sc7179, Santa Cruz). Secondary antibodies were peroxidase-conjugated (Santa Cruz) and used for chemiluminescence using ECL reagents (Amersham, GE Healthcare, Piscataway, NJ, USA).
[0172] 1.2. Treatment of MPM and normal commercial mesothelial cell (HMC)...
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