A Ruthenium Catalyst with Unprecedented Effectiveness for the Coupling Cyclization of γ‑Amino Alcohols and Secondary Alcohols

The ruthenium complex (8-(2-diphenylphosphinoethyl)­aminotrihydroquinolinyl)­(carbonyl)­(hydrido)­ruthenium chloride exhibited extremely high efficiency toward the coupling cyclization of γ-amino alcohols with secondary alcohols. The corresponding products, pyridine or quinoline derivatives, are obtained in good to high isolated yields. On comparison with literature catalysts whose noble-metal loading with respect to γ-amino alcohols reached 0.5–1.0 mol % for Ru and a record lowest of 0.04 mol % for Ir, the current catalyst achieves the same efficiency with a loading of 0.025 mol % for Ru. The mechanism of acceptorless dehydrogenative condensation (ADC) was proposed on the basis of DFT calculations; in addition, the reactive intermediates were determined by GC-MS, NMR, and single-crystal X-ray diffraction. The catalytic process is potentially suitable for industrial applications

Efficient acceptorless dehydrogenation of secondary alcohols to ketones mediated by a PNN-Ru(II) catalyst

Four types of ruthenium(II) complexes, [fac-PNN]RuH(PPh3)(CO) (A), [fac-PNHN]RuH(η1-BH4)(CO) (B), [fac-PNHN]RuCl2(PPh3) (C) and [fac-PNHN]RuH(η1-BH4)(PPh3) (D) (where PNHN and PNN are N-(2-(diphenylphosphino)ethyl)-5,6,7,8-tetrahydroquinoline-8-amine and its deprotonated derivative), have been synthesized and assessed as catalysts for the acceptorless dehydrogenation of secondary alcohols to afford ketones. It was found that C, in combination with t-BuOK, proved the most effective and versatile catalyst allowing aromatic-, aliphatic- and cycloalkyl-containing alcohols to be efficiently converted to their corresponding ketones with particularly high values of TON achievable. Furthermore, the mechanism for this PNN-Ru mediated process been proposed on the basis of a number of intermediates that have been characterized by EI-MS and NMR spectroscopy. These catalysts show great potential for applications in atom-economic synthesis as well as in the development of organic hydride-based hydrogen storage systems

HepG2 k cells express elevated level of VEGFA.

<p>(A) Expression of p-Akt, HIF-1α and VEGFA in parental HepG2 and HepG2 k cells were detected by western blot analysis. (B) The mRNA expression of VEGFA in parental HepG2 and HepG2 k cells was assayed using real time PCR. *, <i>P</i><0.05. (C) VEGFA concentration in conditioned media from parental HepG2 and HepG2 k cells was measured by ELISA analysis. **, <i>P</i><0.01. (D) The expression of HIF-1α and VEGFA in parental HepG2 and HepG2 k cells treated with or without LY294002 (20 µM) for 24 h were analyzed by western blot analysis. (E) VEGFA concentration in conditioned media was detected by ELISA analysis. ***, <i>P</i><0.001 versus parental HepG2 cells control respectively; ^###, <i>P</i><0.001 versus HepG2 k cells; ns, no significance.</p

The enhanced pro-angiogenic ability of HepG2 k is in a HIF-1α/VEGFA dependent manner <i>in vitro</i>.

<p>(A) HUVECs were treated with conditioned media from parental HepG2 or HepG2 k cells with addition of bevacizumab (0.5 mg/ml) or control IgG for 24 h. Cell viability were quantified by MTT assay. (B–C) HUVEC migration <i>in vitro</i> in response to conditioned media from HepG2 k or parental HepG2 cells with addition of bevacizumab (0.5 mg/ml) or control IgG was assayed after 12 h. The number of migrated cells was quantified by counting 10 random fields at ×100 magnification. (D–E) HUVEC tube formation in response to conditioned media from HepG2 k or parental HepG2 cells with addition of bevacizumab (0.5 mg/ml) or control IgG after 20 h was assayed. The length of tube was evaluated by counting 10 random fields at ×100 magnification. (F–H) The parental HepG2 and HepG2 k cells were treated with or without VEGFA siRNA or 5 µM YC-1, and the conditioned media was collected. The effect of various conditioned media on HUVEC 24 h viability was quantified by MTT assay. The effect of various conditioned media on HUVEC migration for 12 h was assayed. The number of migrated cells was quantified by counting 10 random fields at ×100 magnification. The effect of various conditioned media on HUVEC tube formation for 20 h was assayed. The tube length was evaluated by counting 10 random fields at ×100 magnification. *, <i>P</i><0.05, ***, <i>P</i><0.001 versus parental HepG2 cell control; ^##, <i>P</i><0.01, ^###, <i>P</i><0.001 versus HepG2 k cells control; ns, no significance.</p

The viability of HepG2 cells and sublines derived from HepG2 cells after hyperthermia.

<p>(A) HepG2 cells were cultured after 47°C heat treatment. The 24 h, 48 h and 72 h cell viability of HepG2 cells with or without 47°C heat treatment were measured using MTT assay. (B) Twenty-four sublines were established after 47°C heat treatment for 10 min as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0037266#s2" target="_blank">method</a>. The 24 h, 48 h, and 72 h viability was evaluated by MTT assay after 24 sublines were established. par, parental HepG2 cells; a–x, sublines derived from the HepG2 cells. (C) The 24 h, 48 h and 72 h viability of representative sublines of HepG2 cells were evaluated by MTT assay. (D) Parental HepG2 and HepG2 k cells were treated with 49°C or 50°C 10 min. The 4 h, 12 h, 24 h and 48 h cell viability were measured by MTT assay. *, <i>P</i><0.05; **, <i>P</i><0.01; ***, <i>P</i> <0.001. Data are the representative results of three independent experiments. The coefficients of variation (CV) of all assays were shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0037266#pone.0037266.s001" target="_blank">Supporting Information S1</a>.</p

YC-1 and VEGFA siRNA inhibit HepG2 cell viability.

<p>The parental HepG2 and HepG2 k cells were treated with or without YC-1(5 µM) or transfected with or without VEGFA siRNA. (A) The expression of HIF-1α and VEGFA was detected by western blot analysis. (B) VEGFA concentration in conditioned media was detected by ELISA analysis. **, <i>P</i><0.01, ***, <i>P</i><0.001 versus parental HepG2 cells control respectively; ^###, <i>P</i><0.001 versus HepG2 k cells. (C) Parental HepG2 and HepG2 k cells were transfected with or without VEGFA siRNA for 24 h, 48 h, and 72 h, cell viability was measured by MTT assay. ***, <i>P</i><0.01; ns, no significance. (D) Parental HepG2 and HepG2 k cells were treated with or without YC-1 (5 µM) for 24 h, 48 h, and 72 h, cell viability was measured by MTT assay. ***, <i>P</i><0.001; ns, no significance.</p

Bevacizumab impaired the tumor growth and angiogenesis in tumor-bearing mice <i>in vivo</i>.

<p>2×10⁶ HepG2 k or parental HepG2 cells in cells in 200 µl phosphate buffered saline were injected by subcutaneous injection to obtain s. c. tumors. (A) Average tumor volume is shown for the HepG2 tumors. Parental HepG2 tumor with control IgG (n = 5), Parental HepG2 tumor with Bevacizumab (n = 5), HepG2 k tumor with control IgG (n = 5) and HepG2 k tumor with Bevacizumab (n = 5). **, <i>P</i><0.01. (B) Mice were killed after 28 days implantation and the tumor tissues were removed and weighed. *, <i>P</i><0.05. (C–D) Twenty-eight days after implantation, the numbers of new microvessels marked with CD34 (arrow heads) in the subcutaneous tumors were quantified by performing new vessel counts of 10 random fields at ×400 magnification. ^*, <i>P</i><0.05, ^**, <i>P</i><0.01 versus parental HepG2 tumor respectively; ^###, <i>P</i><0.001 versus HepG2 k tumor. (E) Immunohistochemistry analysis of the expression of VEGF in implanted tumors.</p

Additional file 1: of 15-oxoeicosatetraenoic acid mediates monocyte adhesion to endothelial cell

The details of patients are provided as follow. (DOCX 32 kb

Additional file 1: of Myeloperoxidase-oxidized high density lipoprotein impairs atherosclerotic plaque stability by inhibiting smooth muscle cell migration

Supplemental methods and supplemental results. (DOC 125 kb

Expression of SP-A2 wild-type and mutant protein and mRNA in CHO-K1 cells.

<p>(<b>A</b>) SP-A2 schematic. SP-A2 is tagged with V5 epitope at amino acid position 21 follow the signal sequence, four functional domains are indicated, and various mutations are also listed. <b>CRD</b>, carbohydrate recognition domain. (<b>B</b>) CHO-K1 cells were transiently transfected with vector (pcDNA3.0), V5-tagged SP-A2 wild-type, G231V, F198S and Q223K variants full-length cDNA plasmids. Forty-eight hours after transfection, equal amounts of total protein from cell lysate or media were subjected to SDS-PAGE and followed by immunoblot analysis using a monoclonal antibody that recognizes the V5 epitope. (<b>C</b>) Stably expressing vector, V5-tagged SP-A2 wild-type, G231V, F198S and Q223K variants in CHO-K1 cells. 5×10⁵ cells/well were seeded and cultured in 6-well plate for 48 h, cell lysate and medium were collected and analyzed by SDS-PAGE and western blotting. (<b>D</b>) CHO-K1 cells were transiently transfected with plasmids pcDNA3.0, V5-tagged SP-A2 wild-type, G231V, F198S and Q223K variants. Forty-eight hours later, total RNA was extracted and RT-PCR was performed using primers specific to SP-A2 and GAPDH gene.</p