Mn(OAc)₃‑Promoted Oxidative C_sp³–P Bond Formation through C_sp²–C_sp² and P–H Bond Cleavage: Access to β‑Ketophosphonates

The Mn­(OAc)₃-promoted oxidative phosphonylation of <i>N</i>,<i>N</i>-dimethylenaminones with H-phosphonates, involving a chemo- and regioselective C_sp²–C_sp² bond cleavage and C_sp³–P bond formation in one step, provided successfully functionalized β-ketophosphonates under mild reaction conditions. Oxidative C_sp³–H/P–H cross-coupling reactions via C_sp³–C­(CO) bond cleavage and mechanistic studies are conducted preliminarily, and a possible mechanism is proposed. This novel method proceeds in good to excellent yields, shows operational simplicity, broad substrate scope, and large-scale preparation

Layer-by-Layer Assembly of Polysaccharide Films with Self-Healing and Antifogging Properties for Food Packaging Applications

Self-healable, transparent, and antifogging polysaccharide films composed of acrylamide-modified chitosan (AMCS) and alginate aldehyde (ADA) were fabricated via layer-by-layer (LBL) assembly. The Schiff base linkage formed between amino groups of AMCS and aldehyde groups of ADA was used to construct the films. Fourier transform infrared spectroscopy and X-ray photoelectron spectra revealed that the films were cross-linked through Schiff base bonds. Ultraviolet–visible spectroscopy and field emission scanning electron microscope characterizations demonstrated that the films exhibited linear growth during the LBL process. The films showed a repetitive self-healing property, and the repeated damage-healing of the films was thickness-dependent. The films can heal the scratches that penetrated to the underlying substrates. Besides the excellent self-healing property, the films showed an antifogging property due to the hydrophilic nature of the two polysaccharides. The antifogging ability of the film was thickness-dependent. Also, the films could regain their transmittance and fog-resistant property after mechanical abrasion due to their self-healing capability. These self-healable and fog-resistant polymeric films have potential applications in food packaging

Generation and verification of the TERT CKO mice.

<p>(A) Schematic gene-targeting map of TERT gene. The construction of TERT floxed and TERT CKO alleles are shown before and after tamoxifen treatment. TERT gene exon 1–2 was floxed after recombination between WT and targeted alleles. Primers pairs P1/P2 and P3/P5 were used to detect floxed TERT alleles. Primer pair P1/P5 was used to detected the TERT gene excision by Cre. (B) Representative Southern blot analysis. The ES clone with homologous recombination was digested with the restriction enzyme Bsr GI followed by Southern blotting with Dig-labeled 5’ probe as shown in (A). The detected WT and targeted alleles are 14.58 and 7.3 kb, respectively. (C) PCR genotyping using genomic DNA from mouse tails. The PCR fragments for WT was 154bp, for TERT fl/+ they were 154 and 343 bp, for TERT <i>fl/fl</i> it was 343 bp, and for TERT CKO it was 215bp after tamoxifen-induced Cre excision. “Ladder” referred to 100 bp DNA ladder.</p

Effect of TERT overexpression on α-SMA expression.

<p>BJ and BJ 5ta fibroblasts were plated in 6-well plates. The cells were starved with DMEM supplemented with 0.5% FBS for 4 hours before TGF-β1 treatment for an additional 24 for mRNA or 48 hours for protein analysis. (A) BJ and BJ5ta cells were analyzed for TERT mRNA by qRT-PCR. n = 3. *, P < 0.05. (B) The cell lysates were harvested in RIPA buffer, and analyzed for α-SMA protein expression by Western blotting. A representative blot was shown. The GAPDH was used as internal control.</p

TERT and telomerase in floxed TERT vs WT mice.

<p>Total RNA or protein lysates were prepared from the lung tissues and MLF from BLM or PBS-injected TERT <i>fl/fl</i> or WT mice. TERT gene expression was analyzed by qRT-PCR and expressed as 2^-ΔΔCT (n = 3) in (A), and the telomerase activities were detected by TRAP-ELISA kit and expressed as fold change over their PBS control, respectively (B). n = 3. *, P < 0.05.</p

The impairment of pulmonary fibrosis in TERT CKO mice.

<p>(A) The MLF were isolated from TERT CKO or WT mice at 21 days after BLM injection, and analyzed for TERT mRNA by qRT-PCR. The expression was expressed as the fold change of the level in PBS-treated WT MLF. n = 3–5 mice per group. (B) The lungs from TERT CKO and control mice were homogenized at day 21 after BLM treatment, and measured for whole lung collagen content by HYP assay. n = 3–5 mice per group. (C) Lung tissue RNA extracted from the indicated murine strains was also analyzed for type I collagen mRNA by qRT-PCR. n = 3–5 mice per group. (D) Lung tissue lysates were prepared by RIPA buffer, and analyzed for α-SMA protein in the indicated murine strain by Western blotting (top panel). Quantitative data was normalized by the internal control GAPDH, and shown as the percentage of the GAPDH signals (bottom panel). n = 3–4 mice per group. (E) Representative H & E stained lung tissue sections at day 21after BLM treatment are shown. Original magnification × 20. *, P < 0.05.</p

The in vitro excision of MLF TERT by Cre activation.

<p>The MLF were isolated from TERT <i>fl/fl</i> mice, and then transduced with 100 MOI of AdCre vector or AdGFP control vector. Six days after transduction, TERT mRNA (A) and telomerase (B) were analyzed, respectively, as described in the legend of <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0142547#pone.0142547.g002" target="_blank">Fig 2</a>. n = 3. (C) The MLF from TERT <i>fl/fl</i> or TERT <i>fl/fl</i>/,Cre+/- mice were treated with 5 μM of 4-OHT in vitro at the same time for 6 days, and TERT mRNA was analyzed. n = 3. *, P < 0.05.</p

Design, Synthesis, and Structure–Activity Relationship of Novel LSD1 Inhibitors Based on Pyrimidine–Thiourea Hybrids As Potent, Orally Active Antitumor Agents

Histone lysine specific demethylase 1 (LSD1) was reported to be overexpressed in several human cancers and recognized as a promising anticancer drug target. In the current study, we designed and synthesized a novel series of pyrimidine–thiourea hybrids and evaluated their potential LSD1 inhibitory effect. One of the compounds, <b>6b</b>, containing a terminal alkyne appendage, was shown to be the most potent and selective LSD1 inhibitor in vitro and exhibited strong cytotoxicity against LSD1 overexpressed gastric cancer cells. Compound <b>6b</b> also showed marked inhibition of cell migration and invasion as well as significant in vivo tumor suppressing and antimetastasis role, without significant side effects by oral administration. Our findings indicate that the pyrimidine–thiourea-based LSD1 inactivator may serve as a leading compound targeting LSD1 overexpressed cancers