Sofosbuvir Selects for Drug-Resistant Amino Acid Variants in the Zika Virus RNA-Dependent RNA-Polymerase Complex In Vitro

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.     

Test Case Generation as an AI Planning Problem

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.     

Taking advantage of collective knowledge in emergency response systems

When large groups work on a theme, they have the potential to produce a lot of useful knowledge, regardless of whether they are acting in a coordinated manner or individually. Spontaneously generated information has received much attention in recent years, as organizations and businesses discover the power of crowds. New technologies, such as blogs, Twitter, wikis, photo sharing, collaborative tagging and social networking sites, enable the creation and dissemination of content in a relatively simple way. As a result, the aggregate body of knowledge is growing at an accelerated rate. Many organizations are looking for ways to harness this power, which is being called collective intelligence. Research has shown that it is possible to obtain high quality results from collectively produced work.In this paper, we consider the domain of emergency response. Research has shown that individuals respond quickly and massively to emergencies, and that they try to help with the situation. Thus, it seems like a logical step to attempt to harness collective knowledge for emergency management. Disaster relief groups and field command frequently suffer from lack of up to date information, which may be critical in a rapidly evolving situation. Some of this information could be generated by the crowd at large, enabling more effective response to the situation. In this paper, we discuss the possibilities for the introduction of collective knowledge in disaster relief and present architecture and examples of how this could be accomplished.     

The Effects Of Humic Substances On Pinus Callus Are Reversed By 2,4-Dichlorophenoxyacetic Acid

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 (M_r) > 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.     

Minimum Setup Minimum Aberration Two‐level Split‐plot Type Designs for Physical Prototype Testing

Although new technologies allow for less effort in prototyping, physical testing still remains an important step in the product development cycle. Well‐planned experiments are useful to guide the decision‐making process. During the design of an experiment, one of the challenges is to balance limited resources and system constraints to obtain useful information. It is common that prototypes are composed of several parts, with some parts more difficult to assemble than others. And, usually, there is only one piece available of each part type and a large number of different setups. Under these conditions, designs with randomization restrictions become attractive approaches. Considering this scenario, a new and additional criterion, minimum setup, to construct split‐plot type designs is presented. Designs with the minimum number of setups of the more difficult parts, which are especially useful for screening purposes in physical prototype testing, are discussed. The use of the proposed criterion combined with minimum aberration for selecting a regular design is shown through a real application in testing car prototypes. As a tool to practitioners, catalogs of selected 32‐run minimum setup minimum aberration split‐split‐plot and split‐split‐split‐plot designs are presented. More complete catalogs are available as Supporting information. Copyright © 2015 John Wiley & Sons, Ltd.     

BOO: Behavior-oriented ontology to describe participant dynamics in collocated design meetings

Facilitating meetings is not an easy task. To assist the facilitator, we have been designing intelligent support systems, which can help contextual sensemaking, decision making and action. However, these systems are constructed based on behavioral models that provide guidelines to understand participant behaviors. This paper presents an ontology to describe participants’ behaviors in collaborative design meetings and rules that correlate them with the group’s acceptance of the final product. This ontology describes the group dynamics at collocated meetings, using verbal and non-verbal cues of attention shifts and attention maintenance as its basic constructs. The objective of creating this ontology was to better understand face-to-face meetings to eventually help meeting facilitators identify issues that may lead to dissatisfaction with the final product through behavioral cues. The ontology was derived through extensive analysis of a series of engineering design session videos. The design group was composed of experts with similar backgrounds, but working in different divisions of the same company. Different points of view were argued and decisions were made at the end of each meeting. After each meeting, participants were asked to asynchronously commit to the decisions made in the group. Our ontology can be used to identify the factors that lead to an undesired outcome, and now serves as a basis for a new project, which uses rules to support design meetings, improve final artifact acceptance and reduce rework. Our conclusions point out correlations between designers’ behaviors and future artifact acceptance and actions that interrupt or bring back group attention. The ontology was validated through application to other meeting situations. These findings may guide software developers in the creation of tools to support group design, and may be applied by an intelligent system.     

Conversion of a benign lymphoepithelial salivary gland lesion to lymphocytic lymphoma during dilantin therapy: correlation with dilantin-induced lymphocyte transformation in vitro

A patient with a classical salivary gland benign lymphoepithelial lesion (BLL) that converted to a lymphoblastic lymphoma (LSA) localized to the salivary glands is described. The malignant transformation of the BLL was preceded by Dilantin anticonvulsant therapy, and in vitro tests subsequent to the development of LSA demonstrated positive Dilantin-induced lymphocyte transformation. The lymphoma was treated successfully by local irradiation, chemotherapy, and discontinuation of Dilantin. The case illustrates the "prelymphomatous" nature of BLL in certain patients, as well as the possible potential danger of Dilantin and other lymphoid-stimulating drugs in discussed. Further study of drugs capable lymphocyte transformation in patients with prelymphomatous disorders is warranted.     

A Phenylacetamide Resveratrol Derivative Exerts Inhibitory Effects on Breast Cancer Cell Growth

Resveratrol (RSV) is a natural compound that displays several pharmacological properties, including anti-cancer actions. However, its clinical application is limited because of its low solubility and bioavailability. Here, the antiproliferative and anti-inflammatory activity of a series of phenylacetamide RSV derivatives has been evaluated in several cancer cell lines. These derivatives contain a monosubstituted aromatic ring that could mimic the RSV phenolic nucleus and a longer flexible chain that could confer a better stability and bioavailability than RSV. Using MTT assay, we demonstrated that most derivatives exerted antiproliferative effects in almost all of the cancer cell lines tested. Among them, derivative 2, that showed greater bioavailability than RSV, was the most active, particularly against estrogen receptor positive (ER+) MCF7 and estrogen receptor negative (ER-) MDA-MB231 breast cancer cell lines. Moreover, we demonstrated that these derivatives, particularly derivative 2, were able to inhibit NO and ROS synthesis and PGE2 secretion in lipopolysaccharide (LPS)-activated U937 human monocytic cells (derived from a histiocytoma). In order to define the molecular mechanisms underlying the antiproliferative effects of derivative 2, we found that it determined cell cycle arrest at the G1 phase, modified the expression of cell cycle regulatory proteins, and ultimately triggered apoptotic cell death in both breast cancer cell lines. Taken together, these results highlight the studied RSV derivatives, particularly derivative 2, as promising tools for the development of new and more bioavailable derivatives useful in the treatment of breast cancer.     

Optimization of the spark plasma sintering processing parameters affecting the properties of polyimide

The present study deals with the optimization of polyimide (PI) mechanical properties, obtained by Spark Plasma Sintering (SPS), by using a method combining Design of Experiments (DOE) with physical, structural, and mechanical characterizations. The effects of SPS parameters such as temperature, pressure, dwell time, and cooling rate on the density, mechanical properties, and structure of PI were investigated. The experimental results revealed that the mechanical properties of the material were optimized by raising the sintering temperature up to 350°C. The optimized SPS processing parameters were a temperature of 350°C, a pressure of 40 MPa, and a dwell time of 5 min. Under these conditions, a relative density of 99.6% was reached within only a few minutes. The corresponding mechanical properties consisted of Young's modulus of 3.43 GPa, a Shore D hardness of 87.3, and a compressive strength of 738 MPa for a maximum compressive strain of 61.8%. Moreover, when working at 320°C and at 100 MPa, an increase in the dwell time was necessary to enhance the properties. Contrary to the other parameters, the cooling rate appeared to be a non‐significant parameter. Finally, correlations between the PI structure and the mechanical properties were made to demonstrate the densification mechanisms. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015 , 132, 41542.