Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Standardized Extract (HemoHIM) Protects against Scopolamine-Induced Amnesia in a Murine Model

doi: 10.1155/2021/8884243

Figure Lengend Snippet: The effects of HemoHIM on (a) Ach concentration and (b) AchE activity in the hippocampus. Ach levels ( n = five per group) and AchE ( n = 6 per group) in the hippocampus were determined using the ELISA kit. The data are expressed as the mean ± S.E.M. Significant difference from control group (### P < 0.001) and from scopolamine-treated group ( ∗∗∗ P < 0.001, ∗ P < 0.05).

Article Snippet: Acetylcholine ELISA kit and acetylcholinesterase ELISA kit were purchased from Elabscience Biotechnology Co. (Houston, TX, USA).

Techniques: Concentration Assay, Activity Assay, Enzyme-linked Immunosorbent Assay, Control

Journal: International Journal of Molecular Sciences

Article Title: Effect of Cholesterol on the Organic Cation Transporter OCTN1 (SLC22A4)

doi: 10.3390/ijms21031091

Figure Lengend Snippet: Effect of increasing cholesterol (CHS) concentrations on OCTN1 mediated transport of TEA and acetylcholine. The recombinant OCTN1 was reconstituted in liposomes as described in . except that ( a and b ) proteoliposomes batches used for each time course experiment were reconstituted in the presence of 0 (○), 2.1 (●), 4.2 (□), 8.3 (■) % CHS (corresponding to 0, 21, 42 and 83 µg CHS/mg total lipids, respectively). ( a ) Transport was started adding 0.1 mM [ 14 C]-TEA ( b ) or 0.1 mM [ 3 H]-acetylcholine, at time zero to proteoliposomes and stopped at the indicated times as described in . Time course data were interpolated in a first order rate equation. The values are means ± SD from three different experiments. ( c ) Transport rates of [ 14 C]-TEA (○) or [ 3 H]-acetylcholine (●) uptake, calculated as K x transport at equilibrium from ( a , b ), reported as function of the % CHS.

Article Snippet: Amberlite XAD-4, egg yolk phospholipids (3-sn-phosphatidylcoline 60%), CHS, Triton-X100, Sephadex G-75, TEA and acetylcholine were purchased from Merck Life Science s.r.l., 20149 Milano, Italy; tetraethylammonium bromide [ethyl-1- 14 C] was from ARC (American Radiolabeled Chemicals, St. Louis, MO 63146 USA); acetylcholine iodide [acetyl- 3 H] was from Perkin-Elmer Italia S.p.A. 20126 Milano, MI, Italy.

Techniques: Recombinant, Liposomes

Journal: International Journal of Molecular Sciences

Article Title: Effect of Cholesterol on the Organic Cation Transporter OCTN1 (SLC22A4)

doi: 10.3390/ijms21031091

Figure Lengend Snippet: Effect of CHS on regulation of OCTN1 by ATP or NaCl. The recombinant OCTN1 was reconstituted in liposomes as described in . except that in the presence of 8.3% CHS corresponding to 83 µg CHS/mg total lipids and ( a ) in the presence of the indicated ATP concentrations. Transport was started adding 0.1 mM [ 14 C]-TEA (white column, ○) or [ 3 H]-acetylcholine (grey column, □) at time zero to proteoliposomes. ( b ) Increasing NaCl extraliposomal concentration were added together to the radioactive substrates. The transport reaction was stopped at 20 min. Percent residual activity with respect to the control data were interpolated in an IC 50 equation (Dose-response curves). The values are means ± SD from three experiments. Significantly different for * p < 0.01 as calculated from Student’s t test analysis.

Article Snippet: Amberlite XAD-4, egg yolk phospholipids (3-sn-phosphatidylcoline 60%), CHS, Triton-X100, Sephadex G-75, TEA and acetylcholine were purchased from Merck Life Science s.r.l., 20149 Milano, Italy; tetraethylammonium bromide [ethyl-1- 14 C] was from ARC (American Radiolabeled Chemicals, St. Louis, MO 63146 USA); acetylcholine iodide [acetyl- 3 H] was from Perkin-Elmer Italia S.p.A. 20126 Milano, MI, Italy.

Techniques: Recombinant, Liposomes, Concentration Assay, Activity Assay, Control

Journal: International Journal of Molecular Sciences

Article Title: Effect of Cholesterol on the Organic Cation Transporter OCTN1 (SLC22A4)

doi: 10.3390/ijms21031091

Figure Lengend Snippet: Effect of hemicholinium and TEBA on the OCTN1 activity measured in the presence or absence of CHS. The recombinant OCTN1 was reconstituted in liposomes as described in . except that in the absence (○) or in the presence (●) of 8.3% CHS corresponding to 83 µg CHS/mg total lipids. ( a ) Transport was started adding 0.1 mM [ 3 H]-acetylcholine at time zero to proteoliposomes in the presence of increasing external hemicholinium concentrations (0-0.1-0.3-0.5-0.8-1-1.5-2.6 mM) or ( b ) adding 0.1 mM [ 14 C]-TEA in the presence of increasing external TEBA concentrations (0-0.5-1-2.5-5-10-20 mM). The transport reaction was stopped at 20 min. Percent residual activity with respect to the control data were interpolated in an IC 50 equation (Dose-response curves). The values are means ± SD from three experiments.

Article Snippet: Amberlite XAD-4, egg yolk phospholipids (3-sn-phosphatidylcoline 60%), CHS, Triton-X100, Sephadex G-75, TEA and acetylcholine were purchased from Merck Life Science s.r.l., 20149 Milano, Italy; tetraethylammonium bromide [ethyl-1- 14 C] was from ARC (American Radiolabeled Chemicals, St. Louis, MO 63146 USA); acetylcholine iodide [acetyl- 3 H] was from Perkin-Elmer Italia S.p.A. 20126 Milano, MI, Italy.

Techniques: Activity Assay, Recombinant, Liposomes, Control

Journal: International Journal of Molecular Sciences

Article Title: Effect of Cholesterol on the Organic Cation Transporter OCTN1 (SLC22A4)

doi: 10.3390/ijms21031091

Figure Lengend Snippet: Effect of CHS on OCTN1 transport kinetics. The recombinant OCTN1 was reconstituted in liposomes as described in . except that in the absence (○) or in the presence (●) of 8.3% CHS corresponding to 83 µg CHS/mg total lipids. The transport rate was measured adding [ 14 C]-TEA ( a ) or [ 3 H]-acetylcholine ( b ) at the indicated increasing concentrations to proteoliposomes. The transport reaction was stopped at 10 min. Data were plotted according to Lineweaver–Burk as reciprocal transport rate vs reciprocal TEA ( a ) or acetylcholine ( b ) concentration. The values are means ± SD from three experiments.

Article Snippet: Amberlite XAD-4, egg yolk phospholipids (3-sn-phosphatidylcoline 60%), CHS, Triton-X100, Sephadex G-75, TEA and acetylcholine were purchased from Merck Life Science s.r.l., 20149 Milano, Italy; tetraethylammonium bromide [ethyl-1- 14 C] was from ARC (American Radiolabeled Chemicals, St. Louis, MO 63146 USA); acetylcholine iodide [acetyl- 3 H] was from Perkin-Elmer Italia S.p.A. 20126 Milano, MI, Italy.

Techniques: Recombinant, Liposomes, Concentration Assay

Journal: MedComm

Article Title: Mas‐Related G Protein‐Coupled Receptor Member D Sustains Hypertension

doi: 10.1002/mco2.70706

Figure Lengend Snippet: Effect of MrgD on blood pressure. (A) Flow chart of Ad‐MrgD‐treated SD rats; (B–D) the SBP, DBP, and MAP of Ad‐MrgD‐treated SD rats, n = 8 biological replicates per group. (E) Flow chart of MrgD shRNA‐treated SHR rats; (F–H) the SBP, DBP, and MAP of MrgD shRNA‐treated SHR rats, n = 8 biological replicates per group. (I) Flow chart of Ang II‐treated MrgD KO mice; (J–L) the SBP, DBP, and MAP of Ang II‐treated MrgD KO mice, n = 8 biological replicates per group. (M) Representative images of endothelial‐dependent (PFG2α and Ach) vasodilation and vasoconstriction responses. (N) Contraction effect curve of PGF2α‐induced mesenteric arteries, n = 6 biological replicates per group; (O) diastolic effect curve of Ach‐induced mesenteric arteries, n = 6 biological replicates per group. (P) Representative images of non‐endothelial‐dependent (NE and SNP) vasodilation and vasoconstriction responses. (Q) Contraction effect curve of NE‐induced mesenteric arteries, n = 6 biological replicates per group; (R) diastolic effect curve of SNP‐induced mesenteric arteries, n = 6 biological replicates per group. The data were expressed as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Subsequently, vasoconstrictive responses were elicited by cumulative concentrations of PGF2α (Sigma, CAS 4510‐16‐1) or NE (MCE, HY‐13715) over a concentration range of 10 −9 to 10 −4 M. Endothelium‐dependent and ‐independent vasorelaxation were assessed with Ach (MCE, HY‐B0282) and SNP (MCE, HY‐B0564), respectively, using the same concentration range (10 −9 to 10 −4 M).

Techniques: shRNA

Journal: MedComm

Article Title: Mas‐Related G Protein‐Coupled Receptor Member D Sustains Hypertension

doi: 10.1002/mco2.70706

Figure Lengend Snippet: Risperidone alleviated hypertension and remodeling of the mesenteric artery. (A) The structure of risperidone; (B) the structure of risperidone binding with MrgD; (C–E) risperidone relieved Ang II‐induced hypertension in mice, * vs. Saline and # vs. Ang II, n = 8 biological replicates per group. (F and G) Risperidone reversed Ang II‐induced increases in the mRNA levels of collagen I and TGF‐β in the mesenteric artery of mice, n = 5 biological replicates per group. (H–K) Risperidone reversed Ang II‐induced changes in the protein levels of collagen I, TGF‐β, and SM‐22α in the mesenteric artery of mice, n = 5 biological replicates per group. (L) Representative images of endothelial‐dependent (PFG2α and Ach) and non‐endothelial‐dependent (NE and SNP) vasodilation and vasoconstriction responses; (M and N) risperidone reduced vasorelaxation in the mesenteric artery of mice treated with Ang II, * vs. Saline and # vs. Ang II, n = 6 biological replicates per group. (O and P) Risperidone reduced contraction in the mesenteric artery of mice treated with Ang II, * vs. Saline and # vs. Ang II, n = 6 biological replicates per group. The data were expressed as mean ± SEM. */# p < 0.05, **/## p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Subsequently, vasoconstrictive responses were elicited by cumulative concentrations of PGF2α (Sigma, CAS 4510‐16‐1) or NE (MCE, HY‐13715) over a concentration range of 10 −9 to 10 −4 M. Endothelium‐dependent and ‐independent vasorelaxation were assessed with Ach (MCE, HY‐B0282) and SNP (MCE, HY‐B0564), respectively, using the same concentration range (10 −9 to 10 −4 M).

Techniques: Binding Assay, Saline

Journal: International Journal of Molecular Sciences

Article Title: The Human Alpha3 Beta2 Neuronal Nicotinic Acetylcholine Receptor Can Form Two Distinguishable Subtypes

doi: 10.3390/ijms26199506

Figure Lengend Snippet: Sample traces and likely stoichiometries. Injection of hα3 and hβ2 nicotinic acetylcholine receptor (nAChR) mRNA into Xenopus laevis oocytes at ratios of ( a ) 1:5 and ( b ) 5:1 formed functional and kinetically distinguishable receptors. The respective oocytes were perfused with 333 µM and 10 mM acetylcholine (ACh) (EC 85 ) for up to 60 s to characterize desensitization. The percentage of the peak currents remaining after 30 s of sustained ACh application (30 s mark indicated by *) were compared and showed a significant difference between the two injected α3β2 ratios (see also ). Graphical representations of the likely potential stoichiometries are shown below each respective electrophysiological trace (i.e., either α3 (2) + β2 (3) or α3 (3) + β2 (2) ), modeled on what is already published about stoichiometries for α4β2 and the α2β2 nAChRs [ , , , ].

Article Snippet: Plasmids: hα3 (Origene# SC126406) and hβ2 (Origene# SC309051), both in the pCMV6-XL4 plasmid, were purchased from OriGene Technologies Inc., 9620 Medical Center Drive, Suite 200, Rockville, MD, USA, 20850.

Techniques: Injection, Functional Assay