The increased exploitation of microbial sequencing methods has shed light on the high diversity of new microorganisms named Candidate Phyla Radiation (CPR). CPR are mainly detected via 16S rRNA/metabarcoding analyses or metagenomics and are found to be abundant in all environments and present in different human microbiomes. These microbes, characterized by their symbiotic/epiparasitic lifestyle with bacteria, are directly exposed to competition with other microorganisms sharing the same ecological niche. Recently, a rich repertoire of enzymes with antibiotic resistance activity has been found in CPR genomes by using an in silico adapted screening strategy. This reservoir has shown a high prevalence of putative beta-lactamase-encoding genes. We expressed and purified five putative beta-lactamase sequences having the essential domains and functional motifs from class A and class B beta-lactamase. Their enzymatic activities were tested against various beta-lactam substrates using liquid chromatography-mass spectrometry (LC-MS) and showed some beta-lactamase activity even in the presence of a beta-lactamase inhibitor. In addition, ribonuclease activity was demonstrated against RNA that was not inhibited by sulbactam and EDTA. None of these proteins could degrade single- and double-stranded-DNA. This study is the first to express and test putative CPR beta-lactamase protein sequences in vitro. Our findings highlight that the reduced genomes of CPR members harbor sequences encoding for beta-lactamases known to be multifunction hydrolase enzymes. 相似文献
Most in vitro iron mobilization studies from ferritin have been performed in aqueous buffered solutions using a variety of reducing substances. The kinetics of iron mobilization from ferritin in a medium that resembles the complex milieu of cells could dramatically differ from those in aqueous solutions, and to our knowledge, no such studies have been performed. Here, we have studied the kinetics of iron release from ferritin in fresh yeast cell lysates and examined the effect of cellular metabolites on this process. Our results show that iron release from ferritin in buffer is extremely slow compared to cell lysate under identical experimental conditions, suggesting that certain cellular metabolites present in yeast cell lysate facilitate the reductive release of ferric iron from the ferritin core. Using filtration membranes with different molecular weight cut-offs (3, 10, 30, 50, and 100 kDa), we demonstrate that a cellular component >50 kDa is implicated in the reductive release of iron. When the cell lysate was washed three times with buffer, or when NADPH was omitted from the solution, a dramatic decrease in iron mobilization rates was observed. The addition of physiological concentrations of free flavins, such as FMN, FAD, and riboflavin showed about a two-fold increase in the amount of released iron. Notably, all iron release kinetics occurred while the solution oxygen level was still high. Altogether, our results indicate that in addition to ferritin proteolysis, there exists an auxiliary iron reductive mechanism that involves long-range electron transfer reactions facilitated by the ferritin shell. The physiological implications of such iron reductive mechanisms are discussed. 相似文献
Smoking is a risk factor for a variety of deleterious conditions, such as cancer, respiratory disease and cardiovascular disease. Thrombosis is an important and common aspect of several cardiovascular disease states, whose risk is known to be increased by both first- and secondhand smoke. More recently, the residual cigarette smoke that persists after someone has smoked (referred to as thirdhand smoke or THS) has been gaining more attention, since it has been shown that it also negatively affects health. Indeed, we have previously shown that 6-month exposure to THS increases the risk of thrombogenesis. However, neither the time-dependence of THS-induced thrombus formation, nor its sex dependence have been investigated. Thus, in the present study, we investigated these issues in the context of a shorter exposure to THS, specifically 3 months, in male and female mice. We show that the platelets from 3-month THS-exposed mice exhibited enhanced activation by agonists. Moreover, we also show that mice of both sexes exposed to THS have decreased tail bleeding as well as decreased thrombus occlusion time. In terms of the role of sex, intersex disparities in thrombus development and hemostasis as well as in platelet aggregation were, interestingly, observed. Together, our findings show that exposing mice to THS for 3 months is sufficient to predispose them to thrombosis; which seems to be driven, at least in part, by an increased activity in platelets, and that it does not manifest equally in both sexes. 相似文献
Recommendation System is one of such solutions to overcome information overload issues and to identify products most relevant to users and provide suggestions to users for items they might be interested in consuming or elements matching their needs. The significant challenge of several recommendation approaches is that they suggested a huge number of things to the target user. But the exciting items, according to the target user, are seen at the bottom of the recommended list. The proposed approach has improved the quality of recommendations by implementing some of the unique features in the new framework of auto encoder called semi-autoencoder, which contains the rating information as well as some additional information of users. Autoencoder is widely used in the recommender system because it gives the best result for feature extraction, dimensionality reduction, regeneration of data, and a better understanding of the user’s characteristics. The experimental results are compared with some established popular methods using precision, recall, and F-measure evaluation measures. Users generally don’t want to see lots of suggestions. With its six building blocks, the proposed approach gives better performance for the top 10 recommendations compared to other well-known methods.
Spin‐polarized charge transfer at the interface between a ferromagnetic (FM) metal and a molecule can lead to ferromagnetic coupling and to a high spin polarization at room temperature. The magnetic properties of these interfaces can not only alter those of the ferromagnet but can also stabilize molecular spin chains with interesting opportunities toward quantum computing. With the aim to enhance an organic spintronic device's functionality, external control over this spin polarization may thus be achieved by altering the ferromagnet/molecule interface's magnetic properties. To do so, the magnetoelectric properties of an underlying ferroelectric/ferromagnetic interface are utilized. Switching the ferroelectric polarization state of a PbZr0.2Ti0.8O3 (PZT) bottom layer within a PZT/Co/FePc‐based (Pc ‐ phthalocyanine) device alters the X‐ray magnetic circular dichroism of the Fe site within the phthalocyanine molecular top layer. Thus, how to electrically alter the magnetic properties of an interface with high spin polarization at room temperature is demonstrated. This expands electrical control over spin‐polarized FM/molecule interfaces, which is first demonstrated using ferroelectric molecules, to all molecular classes. 相似文献
Measuring network connectivity under grid-based deployment in 3D space is a challenging problem in wireless sensor networks (WSNs). Solving such a problem becomes an even more intricate task with realistic deployment factors such as placement uncertainty and hindrances to wireless communication channels. While much work has been published on optimizing connectivity, only a few papers have addressed such realistic aspects which cause severe connectivity problems in practice. In this paper, we introduce a novel grid-based deployment metric, called Average Connectivity Percentage in order to characterize the deployed network connectivity when sensor placements are subject to random errors around their corresponding grid locations. A generic approach is proposed to assess and evaluate the proposed metric. This generic approach is independent of the grid-shape, random error distributions, and different environment-based channel characteristics. We apply the generic approach in two practical deployment scenarios: the grid-based deployment with bounded uniform errors and with unbounded normal errors. In both cases, the average connectivity percentage is computed numerically and verified via extensive simulations. Based on the numerical results, quantified effects of positioning errors and grid edge length on the average connectivity percentage are outlined. 相似文献
Traditional studies of speaker state focus primarily upon one-stage classification techniques using standard acoustic features. In this article, we investigate multiple novel features and approaches to two recent tasks in speaker state detection: level-of-interest (LOI) detection and intoxication detection. In the task of LOI prediction, we propose a novel Discriminative TFIDF feature to capture important lexical information and a novel Prosodic Event detection approach using AuToBI; we combine these with acoustic features for this task using a new multilevel multistream prediction feedback and similarity-based hierarchical fusion learning approach. Our experimental results outperform published results of all systems in the 2010 Interspeech Paralinguistic Challenge – Affect Subchallenge. In the intoxication detection task, we evaluate the performance of Prosodic Event-based, phone duration-based, phonotactic, and phonetic-spectral based approaches, finding that a combination of the phonotactic and phonetic-spectral approaches achieve significant improvement over the 2011 Interspeech Speaker State Challenge – Intoxication Subchallenge baseline. We discuss our results using these new features and approaches and their implications for future research. 相似文献
In this paper, we address the analysis and recognition of facial expressions in continuous videos. More precisely, we study classifiers performance that exploit head pose independent temporal facial action parameters. These are provided by an appearance-based 3D face tracker that simultaneously provides the 3D head pose and facial actions. The use of such tracker makes the recognition pose- and texture-independent. Two different schemes are studied. The first scheme adopts a dynamic time warping technique for recognizing expressions where training data are given by temporal signatures associated with different universal facial expressions. The second scheme models temporal signatures associated with facial actions with fixed length feature vectors (observations), and uses some machine learning algorithms in order to recognize the displayed expression. Experiments quantified the performance of different schemes. These were carried out on CMU video sequences and home-made video sequences. The results show that the use of dimension reduction techniques on the extracted time series can improve the classification performance. Moreover, these experiments show that the best recognition rate can be above 90%. 相似文献