Radiosensitization of Human Leukemic HL-60 Cells by ATR Kinase Inhibitor (VE-821): Phosphoproteomic Analysis

DNA damaging agents such as ionizing radiation or chemotherapy are frequently used in oncology. DNA damage response (DDR)—triggered by radiation-induced double strand breaks—is orchestrated mainly by three Phosphatidylinositol 3-kinase-related kinases (PIKKs): Ataxia teleangiectasia mutated (ATM), DNA-dependent protein kinase (DNA-PK) and ATM and Rad3-related kinase (ATR). Their activation promotes cell-cycle arrest and facilitates DNA damage repair, resulting in radioresistance. Recently developed specific ATR inhibitor, VE-821 (3-amino-6-(4-(methylsulfonyl)phenyl)-N-phenylpyrazine-2-carboxamide), has been reported to have a significant radio- and chemo-sensitizing effect delimited to cancer cells (largely p53-deficient) without affecting normal cells. In this study, we employed SILAC-based quantitative phosphoproteomics to describe the mechanism of the radiosensitizing effect of VE-821 in human promyelocytic leukemic cells HL-60 (p53-negative). Hydrophilic interaction liquid chromatography (HILIC)-prefractionation with TiO₂-enrichment and nano-liquid chromatography—tandem mass spectrometry (LC-MS/MS) analysis revealed 9834 phosphorylation sites. Proteins with differentially up-/down-regulated phosphorylation were mostly localized in the nucleus and were involved in cellular processes such as DDR, all phases of the cell cycle, and cell division. Moreover, sequence motif analysis revealed significant changes in the activities of kinases involved in these processes. Taken together, our data indicates that ATR kinase has multiple roles in response to DNA damage throughout the cell cycle and that its inhibitor VE-821 is a potent radiosensitizing agent for p53-negative HL-60 cells.     

Neodymium triflate modified nafion composite membrane for reduced alcohol permeability in direct alcohol fuel cell

This paper reports an experimental study on the modification of nafion membrane with neodymium triflate, Nd(SO₃CF₃)₃, a rare earth triflate. The triflate ion resembles nafion in structure and has high lewis acidity, high coordination number, hygroscopic nature and thermal stability. These properties make the neodymium triflate (NdTfO) suitable to improve the performance of NdTfO/nafion membranes in direct alcohol fuel cell by reducing fuel permeability without compromising other properties. The NdTfO/nafion membranes reduced alcohol permeability by nearly 48%. The proton conductivity of 1% NdTfO/nafion was increased by at least 24% as compared to pure cast nafion membrane. The mechanical strength of 1% NdTfO/nafion was higher than that of pure cast nafion. The composite membrane was thermally and chemically stable and has potential for use in direct alcohol fuel cells. A DMFC was developed and its performance was evaluated using the composite membrane, which showed encouraging results.     

Cultivation of human keratinocytes without feeder cells on polymer carriers containing ethoxyethyl methacrylate: in vitro study

The cell/tissue engineering therapy of extensive or chronic skin wounds is a highly topical task of the contemporary medicine. One of possible therapeutic approaches is grafting of in vitro cultured keratinocytes directly to the wound bed, where the cells colonize the wound, proliferate and improve the re-epithelization process. Because the successful cultivation of keratinocytes needs an application of feeder cells, the exclusion of these cells from the cultivation system is highly required. In this study we show a positive influence of 2-ethoxyethyl methacrylate as a component of cultivation support on growth of keratinocytes without feeder cells. Keratinocytes cultured on these surfaces are able to migrate to the model wound bed in vitro, where they form distinct colonies and have a normal differentiation potential.