A facile route to ionic liquids-functionalized ZnO nanorods for the fluorometric sensing of thiabendazole drug
JOURNAL OF MOLECULAR LIQUIDS
Authors: Kaur, Narinder; Raj, Pushap; Singh, Amanpreet; Singh, Narinder; Kim, Deuk Young
Abstract
In this report, we introduce a simple and cost-effective hydrothermal method for the functionalization of ZnO nanorods with three different ionic liquids (ILs (1-3)) to check their sensing response with various drugs including thiabendazole, phenylbutazone, chloramphenicol, neomycin, and theophylline through fluorescence spectroscopy. The rod like-microstructure and hexagonal wurtzite structure of synthesized ZnO nanorods were examined with SEM-EDS and XRD techniques. On the other hand, ionic liquids-functionalized ZnO nanorods (ILs (1-3)/ZnO) exhibited various self-organized (layered-sheet similar to IL1, distorted nanorods similar to IL2, peanut beaded surface similar to IL3/ZnO) structures due to the intercalation of ILs on the surface of ZnO nanorods. Compared to ZnO nanorods, similar to ILs (1-3)/ZnO nanorods showed a blue shift in UV-absorption band which resulted to the variation in optical bandgap values. Ionic liquids-functionalized nanorods samples (ILs (1-3)/ZnO) have been selected for their recognition behaviour with available drug molecules using fluorescence spectroscopic technique. Interestingly, with the addition of thiabendazole drug molecules to two samples (ILs (1,3)/ZnO), a significant response corresponding to the peaks at similar to 350 nm and similar to 357 nm was observed, which led to the development of fluorescent sensors with a detection limit of 12 and 304 nM, respectively. (C) 2018 Elsevier B.V. All rights reserved.
Cytokine-inducible promoters to drive dynamic transgene expression: The "Smart Graft" strategy
XENOTRANSPLANTATION
Authors: Fischer, Andrea; Manske, Katrin; Seissler, Jochen; Wohlleber, Dirk; Simm, Nina; Wolf-van Buerck, Lelia; Knolle, Percy; Schnieke, Angelika; Fischer, Konrad
Abstract
Background Ubiquitous expression of T-cell regulatory transgenes such as the cytotoxic T lymphocyte-associated antigen 4 (CTLA4) or the high-affinity variant LEA29Y improves xeno graft survival. Such donor pigs are however immunocompromised and susceptible to infection. Continous high expression of CTLA4 or LEA29Y in the graft could also compromise the health status of recipients. The novel "Smart Graft" strategy is likely to avoid these problems by controlling the expression of T-cell regulatory transgenes as and when required. Methods Candidate promoters inducible by inflammatory cytokines were identified by in silico screening for potential NF-kappa B binding sites. Basal promoter levels and responsiveness to TNF alpha and IL1ss were quantified by expression of secreted embryonic alkaline phosphatase in cultured cells. Promoters were modified to increase responsiveness by removing regulatory elements or adding SP-1 or NF-kappa B binding sites and again tested in vitro. The most promising promoters were then assessed in vivo. Porcine cells expressing inducible Renilla luciferase constructs were transplanted into immunodeficient NOD-Scid-IL2 receptor gamma(null)(NSG) mice. Following engraftment, the recipient's immune system was reconstituted by splenocyte transfer raising an immune response to the porcine xenograft. The resulting induction of promoter activity was detected by in vivo bioimaging. Results Three human (hTNFAIP1, hVCAM1 and hCCL2), and one porcine promoter (pA20) were chosen for in vitro tests. In all experiments, the semi-synthetic and inducible ELAM promoter as well as the CAG promoter were used as references. In contrast to hTNFAIP1 and hVCAM1 the ELAM, hCCL2 and pA20 promoters showed significant induction after cytokine challenge. The hCCL2 and pA20 promoters were further optimized, resulting in increased responsiveness to TNF alpha and IL1ss. Cytokine-dependent upregulation of promoter activity was tested in vivo, where the ELAM and the optimized hCCL2 promoters showed a 2-fold upregulation, while one of the improved A20 promoters showed almost 10-fold upregulation. Our results also revealed more than 4-fold cytokine inducibility of the CAG promoter. Conclusion This is the first in vivo comparison of existing and newly designed cytokine-inducible promoters. Optimization of promoter structure resulted in almost 10-fold inducibility of promoter activity. Such a rapid and dynamically regulated response to inflammation and cell damage could reduce initial graft rejection, making the "Smart Graft" approach a useful means of modulating the expression of immune regulatory transgenes to avoid deleterious effects on porcine and human health. Expressing transgenes in this fashion could provide a safer organ for transplantation.