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1.
Adele Boccuto Filippo Dragoni Francesca Picarazzi Alessia Lai Carla Della Ventura Carla Veo Federica Giammarino Francesco Saladini Gianguglielmo Zehender Maurizio Zazzi Mattia Mori Ilaria Vicenti 《International journal of molecular sciences》2021,22(5)
The nucleotide analog sofosbuvir, licensed for the treatment of hepatitis C, recently revealed activity against the Zika virus (ZIKV) in vitro and in animal models. However, the ZIKV genetic barrier to sofosbuvir has not yet been characterized. In this study, in vitro selection experiments were performed in infected human hepatoma cell lines. Increasing drug pressure significantly delayed viral breakthrough (p = 0.029). A double mutant in the NS5 gene (V360L/V607I) emerged in 3 independent experiments at 40–80 µM sofosbuvir resulting in a 3.9 ± 0.9-fold half- maximal inhibitory concentration (IC50) shift with respect to the wild type (WT) virus. A triple mutant (C269Y/V360L/V607I), detected in one experiment at 80 µM, conferred a 6.8-fold IC50 shift with respect to the WT. Molecular dynamics simulations confirmed that the double mutant V360L/V607I impacts the binding mode of sofosbuvir, supporting its role in sofosbuvir resistance. Due to the distance from the catalytic site and to the lack of reliable structural data, the contribution of C269Y was not investigated in silico. By a combination of sequence analysis, phenotypic susceptibility testing, and molecular modeling, we characterized a double ZIKV NS5 mutant with decreased sofosbuvir susceptibility. These data add important information to the profile of sofosbuvir as a possible lead for anti-ZIKV drug development. 相似文献
2.
Test Case Generation as an AI Planning Problem 总被引:6,自引:0,他引:6
Adele E. Howe Anneliese von Mayrhauser Richard T. Mraz 《Automated Software Engineering》1997,4(1):77-106
While Artificial Intelligence techniques have been applied to a variety of software engineering applications, the area of automated software testing remains largely unexplored. Yet, test cases for certain types of systems (e.g., those with command language interfaces and transaction based systems) are similar to plans. We have exploited this similarity by constructing an automated test case generator with an AI planning system at its core. We compared the functionality and output of two systems, one based on Software Engineering techniques and the other on planning, for a real application: the StorageTek robot tape library command language. From this, we showed that AI planning is a viable technique for test case generation and that the two approaches are complementary in their capabilities. 相似文献
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The sensitivity to material parameters of surface plasmon oscillations is explored for the design of a prototype system operating at a wavelength of 670 nm, for the simultaneous determination of thickness, complex permittivity and surface roughness of metal films. The apparatus was employed for characterization of gold and silver films with thicknesses within the 10-100 nm range. Thickness data measured with the prototype system were well correlated with those measured on a conventional surface profilometer. The Fourier spectrum of surface irregularities extracted from scattered light, enhanced by surface plasmon oscillations, was used to determine surface roughness parameters for the distinct samples. Those parameters were shown to be representative of typical surface structures observed with the atomic force microscope 相似文献
5.
The regulation of programmed cell death in the developing nervous system involves target-derived survival factors, afferent synaptic activity, and hormone- and cytokine-dependent signaling. Cultured immature cerebellar granule neurons die by apoptosis within several days in vitro unless maintained in depolarizing (high) concentrations of potassium (25 mM K+). Here we report that transforming growth factors (TGF)-beta1, -beta2, and -beta3 accelerate apoptosis of these neurons when maintained in physiological (low) K+ medium (5mM K+) as assessed by measures of viability, quantitative DNA fragmentation, and nuclear morphology. TGF-beta-induced apoptosis of these neurons is not blocked by CNTF and LIF, cytokines that enhance neuronal survival when applied alone, or by IGF-I, which prevents apoptosis upon potassium withdrawal. In contrast, neurons that differentiate in high K+ medium for several days in vitro acquire resistance to TGF-beta-mediated cell death. Granule neurons maintained in either low or high K+ medium produce latent, but not bioactive, TGF-beta1 and -beta2. Because neutralizing TGF-beta antibodies fail to augment survival of low K+ neurons, the cerebellar neurons are apparently unable to activate latent TGF-beta. Thus, apoptosis of low K+ neurons is not attributable to endogenous production of TGF-beta. Taken together, our data suggest that TGF-beta may limit the expansion of postmitotic neuronal precursor populations by promoting their apoptosis but may support survival of those neurons that have maturated, differentiated, and established supportive synaptic connectivity. 相似文献
6.
The reversal of humic matter-induced inhibition of callus growth and metabolism by 2,4-dichlorophenoxyacetic acid (2,4-D) was studied in Pinus laricio. Two forest humic fractions (relative molecular mass (Mr) > 3500), derived from soil under Fagus sylvatica (Fs) and Abies alba (Aa) plantation, were used. Pinus laricio callus was grown for a subculture period (4 weeks) on Basal Murashige and Skoog (MS) medium plus forest humic matters (Fs or Aa), at a concentration of 1 mg C/l, and then was transferred, for an additional four weeks, to a MS medium culture without humic matter, but with different hormones: indole-3-acetic acid (IAA, 2 mg/1) or 2,4-dichlorophenoxyacetic acid (2,4-D, 0.5 mg/1) and/or 6-benzylaminopurine (BAP, 0.25 mg/1). Growth of calluse, glucose, fructose, and sucrose contents, and activities of soluble and bound invertases, glucokinase, phosphoglucose isomerase, aldolase, and pyruvate kinase were monitored. The results show a negative effect of humic fractions on callus growth, due to decreased utilization of glucose and fructose, and decreased activities of glycolytic enzymes. The effects are reversible. Substitution of humic fractions with 2,4-D+BAP or 2,4-D is followed by an increase of glycolytic enzyme activities and, consequently, by the utilization of glucose and fructose that induces a restart of growth. In contrast, the inhibitory effects of humic fractions persist when they are substituted with BAP alone, indicating that only the auxin 2,4-D is capable of reversing the negative effects. A possible competitive action on the auxin-binding site between 2,4-D and the chemical structures in the forest humic fractions is suggested. 相似文献
7.
Engineered Multifunctional Albumin‐Decorated Porous Silicon Nanoparticles for FcRn Translocation of Insulin
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João P. Martins Roberto D'Auria Dongfei Liu Flavia Fontana Mónica P. A. Ferreira Alexandra Correia Marianna Kemell Karina Moslova Ermei Mäkilä Jarno Salonen Luca Casettari Jouni Hirvonen Bruno Sarmento Hélder A. Santos 《Small (Weinheim an der Bergstrasse, Germany)》2018,14(27)
The last decade has seen remarkable advances in the development of drug delivery systems as alternative to parenteral injection‐based delivery of insulin. Neonatal Fc receptor (FcRn)‐mediated transcytosis has been recently proposed as a strategy to increase the transport of drugs across the intestinal epithelium. FcRn‐targeted nanoparticles (NPs) could hijack the FcRn transcytotic pathway and cross the epithelial cell layer. In this study, a novel nanoparticulate system for insulin delivery based on porous silicon NPs is proposed. After surface conjugation with albumin and loading with insulin, the NPs are encapsulated into a pH‐responsive polymeric particle by nanoprecipitation. The developed NP formulation shows controlled size and homogeneous size distribution. Transmission electron microscopy (TEM) images show successful encapsulation of the NPs into pH‐sensitive polymeric particles. No insulin release is detected at acidic conditions, but a controlled release profile is observed at intestinal pH. Toxicity studies show high compatibility of the NPs with intestinal cells. In vitro insulin permeation across the intestinal epithelium shows approximately fivefold increase when insulin is loaded into FcRn‐targeted NPs. Overall, these FcRn‐targeted NPs offer a toolbox in the development of targeted therapies for oral delivery of insulin. 相似文献
8.
GP Fontana 《Canadian Metallurgical Quarterly》1998,8(4):871-890
In a short period of time, tremendous progress has been made in the field of minimally invasive cardiac surgery. The goal of true "minimally invasive" cardiac surgery should be a totally thoracoscopic or transvascular procedure that allows a very short hospital stay and prompt patient recovery at acceptable costs. To accomplish this goal, efforts to miniaturize and refine instrumentation/cannulae/visualization and the development of innovative new techniques must be pursued. The concept of performing intracardiac repair of a variety of pathologies on the beating heart is under investigation and will require a new generation of technology, which will include through-blood imaging, such as blood displacement videoscopes, and task-specific instruments. Robotics may offer additional assistance in the performance of complex and fine maneuvers. 相似文献
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10.
Impact of orthorectification and spatial sampling on maximum NDVI composite data in mountain regions 总被引:1,自引:0,他引:1
Fabio M.A. Fontana Alexander P. Trishchenko Yi Luo Stefan Wunderle 《Remote sensing of environment》2009,113(12):2701-2712
Topography and accuracy of image geometric registration significantly affect the quality of satellite data, since pixels are displaced depending on surface elevation and viewing geometry. This effect should be corrected for through the process of accurate image navigation and orthorectification in order to meet the geolocation accuracy for systematic observations specified by the Global Climate Observing System (GCOS) requirements for satellite climate data records. We investigated the impact of orthorectification on the accuracy of maximum Normalized Difference Vegetation Index (NDVI) composite data for a mountain region in north-western Canada at various spatial resolutions (1 km, 4 km, 5 km, and 8 km). Data from AVHRR on board NOAA-11 (1989 and 1990) and NOAA-16 (2001, 2002, and 2003) processed using a system called CAPS (Canadian AVHRR Processing System) for the month of August were considered. Results demonstrate the significant impact of orthorectification on the quality of composite NDVI data in mountainous terrain. Differences between orthorectified and non-orthorectified NDVI composites (ΔNDVI) adopted both large positive and negative values, with the 1% and 99% percentiles of ΔNDVI at 1 km resolution spanning values between − 0.16 < ΔNDVI < 0.09. Differences were generally reduced to smaller numbers for coarser resolution data, but systematic positive biases for non-orthorectified composites were obtained at all spatial resolutions, ranging from 0.02 (1 km) to 0.004 (8 km). Analyzing the power spectra of maximum NDVI composites at 1 km resolution, large differences between orthorectified and non-orthorectified AVHRR data were identified at spatial scales between 4 km and 10 km. Validation of NOAA-16 AVHRR NDVI with MODIS NDVI composites revealed higher correlation coefficients (by up to 0.1) for orthorectified composites relative to the non-orthorectified case. Uncertainties due to the AVHRR Global Area Coverage (GAC) sampling scheme introduce an average positive bias of 0.02 ± 0.03 at maximum NDVI composite level that translates into an average relative bias of 10.6% ± 19.1 for sparsely vegetated mountain regions. This can at least partially explain the systematic average positive biases we observed relative to our results in AVHRR GAC-based composites from the Global Inventory Modeling and Mapping Studies (GIMMS) and Polar Pathfinder (PPF) datasets (0.19 and 0.05, respectively). With regard to the generation of AVHRR long-term climate data records, results suggest that orthorectification should be an integral part of AVHRR pre-processing, since neglecting the terrain displacement effect may lead to important biases and additional noise in time series at various spatial scales. 相似文献