Journal: Molecular pharmaceutics

Article Title: Preclinical Efficacy and Characterization of Candidate Vaccines for Treatment of Opioid Use Disorders Using Clinically Viable Carrier Proteins

doi: 10.1021/acs.molpharmaceut.8b00592

Figure Lengend Snippet: Analytical Assays To Characterize Carrier Proteins and Oxycodone Conjugate Vaccines a

Article Snippet: After stirring for 10 min, native KLH (mcKLH #7653, Thermo Fisher, Rockford, IL, Figures , , , and ), subunit KLH (sKLH, GMP grade, Biosyn, Carlsbad, CA, , and ), TT and CRM 197 (Walvax Biotechnology Co., Kunming, China), and EcoCRM and rTTHc proteins (Fina Biosolutions, LLC., Rockville, MD) were added at a final concentration of 2.8 mg/mL.

Techniques: SDS Page

Journal: Pharmacology Research & Perspectives

Article Title: Orthovanadate-induced vasocontraction is mediated by the activation of Rho-kinase through Src-dependent transactivation of epidermal growth factor receptor

doi: 10.1002/prp2.39

Figure Lengend Snippet: Effects of TAPI-0 and CRM 197 on sodium orthovanadate (OVA)-induced aortic contraction and phosphorylation of epidermal growth factor receptor (EGFR) and myosin phosphatase target subunit 1 (MYPT1) in rat aortic smooth muscle cells (VSMCs). (A) Contractile force was measured 10 min after treatment with OVA (500 μ mol/L) in rings without endothelium. TAPI-0 (10 μ mol/L) and CRM 197 (10 μ g/mL) were added to the bath 15 min before OVA treatment. The contractile force was expressed as a percentage of the maximal force evoked by 40 mmol/L KCl. Data for rings taken from four animals are expressed as mean ± SE * P < 0.05 vs. OVA alone. (B) Phosphorylated EGFR (Tyr-1173), MYPT1 (Thr-853), and the respective total proteins were measured in VSMCs 10 min after treatment with OVA (500 μ mol/L) by western blotting. TAPI-0 (10 μ mol/L), CRM 197 (10 μ g/mL), or vehicle (Veh; 10 μ L of dimethyl sulfoxide) was added to cultures 15 min before treatment with OVA. Bar graphs represent densitometric quantification of the ratios of phosphorylated to total EGFR and MYPT1. Data are presented as the mean ± SE of four independent experiments. * P < 0.05 vs. Veh; Ψ P < 0.05 vs. OVA alone.

Article Snippet: The following inhibitors were purchased from the commercial sources indicated: AG1478 [4-(3-chloroanilino)-6,7-dimethoxyquinazoline], EGFR inhibitor 1 [cyclopropanecarboxylic acid-(3-(6-(3-trifluoromethyl-phenylamino)-pyrimidin-4-ylamino)-phenyl)-amide], hydroxyfasudil [1-(1-hydroxy-5-isoquinolinesulfonyl)homopiperazine], ML-7 [1-(5-iodonaphthalene-1-sulfonyl)homopiperazine], PP2 {[4-amino-3-(4-chlorophenyl)-1-( t -butyl)-1 H -pyrazolo[3,4-d]pyrimidine}, PTP-I, TAPI-0 { N -( R )-[2-(hydroxyaminocarbonyl) methyl]-4-methylpentanoyl- l -naphthylalanyl- l -alanine amide}, and Y-27632 { R -[+]-trans- N -[4-pyridyl]-4-[1-aminoethyl]-cyclohexanecarboxamide} from Merck-Millipore (Tokyo, Japan); Src inhibitor No. 5 {5 [4-(3′-methoxy-6′-chloro-anilino)-6-methoxy-7(morpholino-3-propoxy)-quinazoline]} from Biaffin GmbH & Co KG (Kassel, Germany); CRM 197 (an inactive diphtheria toxin) from WAKO (Osaka, Japan); OVA from Nakarai Tesque (Kyoto, Japan).

Techniques: Western Blot