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Shreepa J. Chauhan Anita Thyagarajan Yanfang Chen Jeffrey B. Travers Ravi P. Sahu 《International journal of molecular sciences》2020,21(22)
Microvesicle particles (MVP) secreted by a variety of cell types in response to reactive oxygen species (ROS)-generating pro-oxidative stressors have been implicated in modifying the cellular responses including the sensitivity to therapeutic agents. Our previous studies have shown that expression of a G-protein coupled, platelet-activating factor-receptor (PAFR) pathway plays critical roles in pro-oxidative stressors-mediated cancer growth and MVP release. As most therapeutic agents act as pro-oxidative stressors, the current studies were designed to determine the role of the PAFR signaling in targeted therapies (i.e., gefitinib and erlotinib)-mediated MVP release and underlying mechanisms using PAFR-expressing human A549 and H1299 non-small cell lung cancer (NSCLC) cell lines. Our studies demonstrate that both gefitinib and erlotinib generate ROS in a dose-dependent manner in a process blocked by antioxidant and PAFR antagonist, verifying their pro-oxidative stressor’s ability, and the role of the PAFR in this effect. We observed that these targeted therapies induce MVP release in a dose- and time-dependent manner, similar to a PAFR-agonist, carbamoyl-PAF (CPAF), and PAFR-independent agonist, phorbol myristate acetate (PMA), used as positive controls. To confirm the PAFR dependency, we demonstrate that siRNA-mediated PAFR knockdown or PAFR antagonist significantly blocked only targeted therapies- and CPAF-mediated but not PMA-induced MVP release. The use of pharmacologic inhibitor strategy suggested the involvement of the lipid ceramide-generating enzyme, acid sphingomyelinase (aSMase) in MVP biogenesis, and observed that regardless of the stimuli used, aSMase inhibition significantly blocked MVP release. As mitogen-activated protein kinase (MAPK; ERK1/2 and p38) pathways crosstalk with PAFR, their inhibition also significantly attenuated targeted therapies-mediated MVP release. These findings indicate that PAFR signaling could be targeted to modify cellular responses of targeted therapies in lung cancer cells. 相似文献
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Dr. Bekir Engin Eser Michal Poborsky Rongrong Dai Dr. Shigenobu Kishino Anita Ljubic Dr. Michiki Takeuchi Prof. Dr. Charlotte Jacobsen Prof. Dr. Jun Ogawa Prof. Dr. Peter Kristensen Prof. Dr. Zheng Guo 《Chembiochem : a European journal of chemical biology》2020,21(4):550-563
Enzymatic conversion of fatty acids (FAs) by fatty acid hydratases (FAHs) presents a green and efficient route for high-value hydroxy fatty acid (HFA) production. However, limited diversity was achieved among HFAs, to date, with respect to chain length and hydroxy position. In this study, two highly similar FAHs from Lactobacillus acidophilus were compared: FA-HY2 has a narrow substrate scope and strict regioselectivity, whereas FA-HY1 utilizes longer chain substrates and hydrates various double-bond positions. It is revealed that three active-site residues play a remarkable role in directing substrate specificity and regioselectivity of hydration. If these residues on FA-HY2 are mutated to the corresponding ones in FA-HY1, a significant expansion of substrate scope and a distinct enhancement in hydration of double bonds towards the ω-end of FAs is observed. A three-residue mutant of FA-HY2 (TM-FA-HY2) displayed an impressive reversal of regioselectivity towards linoleic acid, shifting the ratio of the HFA regioisomers (10-OH/13-OH) from 99:1 to 12:88. Notable changes in regioselectivity were also observed for arachidonic acid and for C18 polyunsaturated fatty acid substrates. In addition, TM-FA-HY2 converted eicosapentaenoic acid into its 12-hydroxy product with high conversion at the preparative scale. Furthermore, it is demonstrated that microalgae are a source of diverse FAs for HFA production. This study paves the way for tailor-made FAH design to enable the production of diverse HFAs for various applications from the polymer industry to medical fields. 相似文献
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Dr. Aashrita Rajagopalan Dr. Markus Schober Anita Emmerstorfer Lucas Hammerer Anna Migglautsch Dr. Birgit Seisser Dr. Silvia. M. Glueck Dr. Frank Niehaus Dr. Juergen Eck Dr. Harald Pichler Prof. Dr. Karl Gruber Prof. Dr. Wolfgang Kroutil 《Chembiochem : a European journal of chemical biology》2013,14(18):2427-2430
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In this study the use of carbon anode dust (CAD), which is the solid residue from aluminum production, as a low-cost adsorbent for the removal of Ni(II) ions from wastewater was investigated. A mechanism of adsorption kinetics and thermodynamics was proposed. In order to investigate the adsorption process of nickel ions on CAD three kinetic models were used: Lagergren's pseudo-first-order model, pseudo-second-order model, and the intra-particle diffusion model. Various thermodynamic parameters, such as the energy of activation (Ea), activation enthalpy (ΔH*), activation entropy (ΔS*), and free energy of activation (ΔG*), were evaluated. Observation of the value of the energy of activation suggests that the process uptake of Ni(II) ions can be described as activated chemisorption. The positive values of enthalpy of activation and free energy of activation, as well as negative values of entropy of activation, mean that the process of removal of Ni(II) ions is endothermic, non-spontaneous, and without structural changes in the solid carbon anode dust particles. 相似文献