Feline coronaviruses (FCoV) exist as 2 biotypes: feline enteric coronavirus (FECV) and feline infectious peritonitis virus (FIPV). FECV causes subclinical infections; FIPV causes feline infectious ...peritonitis (FIP), a systemic and fatal disease. It is thought that mutations in FECV enable infection of macrophages, causing FIP. However, the molecular basis for this biotype switch is unknown. We examined a furin cleavage site in the region between receptor-binding (S1) and fusion (S2) domains of the spike of serotype 1 FCoV. FECV sequences were compared with FIPV sequences. All FECVs had a conserved furin cleavage motif. For FIPV, there was a correlation with the disease and >1 substitution in the S1/S2 motif. Fluorogenic peptide assays confirmed that the substitutions modulate furin cleavage. We document a functionally relevant S1/S2 mutation that arises when FIP develops in a cat. These insights into FIP pathogenesis may be useful in development of diagnostic, prevention, and treatment measures against coronaviruses.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, ODKLJ, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
A critical step in the influenza virus replication cycle is the cleavage activation of the HA precursor. Cleavage activation of influenza HA enables fusion with the host endosome, allowing for ...release of the viral genome into the host cell. To date, studies have determined that HA activation is driven by trypsin-like host cell proteases, as well as yet to be identified bacterial proteases. Although the number of host proteases that can activate HA is growing, there is still uncertainty regarding which secreted proteases are able to support multicycle replication of influenza. In this study, we have determined that the kallikrein-related peptidases 5 and 12 are secreted from the human respiratory tract and have the ability to cleave and activate HA from the H1, H2, and H3 subtypes. Each peptidase appears to have a preference for particular influenza subtypes, with kallikrein 5 cleaving the H1 and H3 subtypes most efficiently and kallikrein 12 cleaving the H1 and H2 subtypes most efficiently. Cleavage analysis using HA cleavage site peptide mimics revealed that the amino acids neighboring the arginine cleavage site affect cleavage efficiency. Additionally, the thrombolytic zymogens plasminogen, urokinase, and plasma kallikrein have all been shown to cleave and activate influenza but are found circulating mainly as inactive precursors. Kallikrein 5 and kallikrein 12 were examined for their ability to activate the thrombolytic zymogens, and both resulted in activation of each zymogen, with kallikrein 12 being a more potent activator. Activation of the thrombolytic zymogens may therefore allow for both direct and indirect activation of the HA of human-adapted influenza viruses by kallikrein 5 and kallikrein 12.
Background: Cleavage of the influenza HA precursor by host proteases is a critical step in the virus life cycle.
Results: The airway-secreted proteases kallikrein 5 and kallikrein 12 were found to cleave and activate influenza HA.
Conclusion: Each peptidase had distinct preferences for particular human-adapted influenza viruses.
Significance: Identification of these HA-cleaving peptidases aids in our understanding of host proteases involved in influenza infection.
Hemagglutinin (HA) is the viral protein that facilitates the entry of influenza viruses into host cells. This protein controls two critical aspects of entry: virus binding and membrane fusion. In ...order for HA to carry out these functions, it must first undergo a priming step, proteolytic cleavage, which renders it fusion competent. Membrane fusion commences from inside the endosome after a drop in lumenal pH and an ensuing conformational change in HA that leads to the hemifusion of the outer membrane leaflets of the virus and endosome, the formation of a stalk between them, followed by pore formation. Thus, the fusion machinery is an excellent target for antiviral compounds, especially those that target the conserved stem region of the protein. However, traditional ensemble fusion assays provide a somewhat limited ability to directly quantify fusion partly due to the inherent averaging of individual fusion events resulting from experimental constraints. Inspired by the gains achieved by single molecule experiments and analysis of stochastic events, recently-developed individual virion imaging techniques and analysis of single fusion events has provided critical information about individual virion behavior, discriminated intermediate fusion steps within a single virion, and allowed the study of the overall population dynamics without the loss of discrete, individual information. In this article, we first start by reviewing the determinants of HA fusogenic activity and the viral entry process, highlight some open questions, and then describe the experimental approaches for assaying fusion that will be useful in developing the most effective therapies in the future.
Volumetric wave-based acoustic simulation relies on the complete solution to the three-dimensional wave equation over a spatial grid. Detailed modeling of sources, however, requires interpolation ...over the grid, which is complicated by the directional character of the source itself. In this paper, a new model of point sources of arbitrary directivity and location with respect to an underlying grid is presented. The model is framed in the spatio-temporal domain directly through the differentiation of Dirac distributions, leading to a spatial Fourier-based approximation strategy. Various approximants are presented, of both separable and nonseparable type, which allow for optimization over a specified wavenumber range. Such approximants are then employed in a finite difference time domain setting, yielding numerical results for sources of various types, which are then compared against exact solutions.
In room acoustics simulation and virtualization applications, accurate wall termination is a perceptually crucial feature. It is particularly important in the setting of wave-based modeling of 3D ...spaces, using methods such as the finite difference time domain method or finite volume time domain method. In this paper, general locally reactive impedance boundary conditions are incorporated into a 3D finite volume time domain formulation, which may be specialized to the various types of finite difference time domain method under fitted boundary termination. Energy methods are used to determine stability conditions for general room geometries, under a large family of nontrivial wall impedances, for finite volume methods over unstructured grids. Simulation results are presented, highlighting in particular the need for unstructured or fitted cells at the room boundary in the case of the accurate simulation of frequency-dependent room mode decay times.
Computer modeling in acoustics allows for the prediction of acoustical defects and the evaluation of potential remediations. In this article, computer modeling is applied to the case of a ...barrel-vaulted sanctuary whose architectural design and construction led to severe flutter echoes along the main aisle, which was later mitigated through acoustical remediations. State-of-the-art geometrical acoustics and wave-based simulations are carried out to analyze the acoustics of this space, with a particular focus on the flutter echoes along the main aisle, before and after remediations. Multi-resolution wavelet and spectrogram analyses are carried out to isolate and characterize flutter echoes within measurements and computer-simulated room impulse responses. Comparisons of simulated responses to measurements are also made in terms of decay times and curves. Simulated room impulse responses from both geometrical acoustics and wave-based methods show evidence of flutter echoes matching measurements, to varying degrees. Time-frequency analyses isolating flutter echoes demonstrate better matches to measurements from wave-based simulated responses, at the cost of longer simulation times than geometrical acoustics simulations. This case study highlights the importance of computer modeling of acoustics in early design phases of architectural planning of worship spaces.
Getting access to administrative health data for research purposes is a difficult and time-consuming process due to increasingly demanding privacy regulations. An alternative method for sharing ...administrative health data would be to share synthetic datasets where the records do not correspond to real individuals, but the patterns and relationships seen in the data are reproduced. This paper assesses the feasibility of generating synthetic administrative health data using a recurrent deep learning model. Our data comes from 120,000 individuals from Alberta Health's administrative health database. We assess how similar our synthetic data is to the real data using utility assessments that assess the structure and general patterns in the data as well as by recreating a specific analysis in the real data commonly applied to this type of administrative health data. We also assess the privacy risks associated with the use of this synthetic dataset. Generic utility assessments that used Hellinger distance to quantify the difference in distributions between real and synthetic datasets for event types (0.027), attributes (mean 0.0417), Markov transition matrices (order 1 mean absolute difference: 0.0896, sd: 0.159; order 2: mean Hellinger distance 0.2195, sd: 0.2724), the Hellinger distance between the joint distributions was 0.352, and the similarity of random cohorts generated from real and synthetic data had a mean Hellinger distance of 0.3 and mean Euclidean distance of 0.064, indicating small differences between the distributions in the real data and the synthetic data. By applying a realistic analysis to both real and synthetic datasets, Cox regression hazard ratios achieved a mean confidence interval overlap of 68% for adjusted hazard ratios among 5 key outcomes of interest, indicating synthetic data produces similar analytic results to real data. The privacy assessment concluded that the attribution disclosure risk associated with this synthetic dataset was substantially less than the typical 0.09 acceptable risk threshold. Based on these metrics our results show that our synthetic data is suitably similar to the real data and could be shared for research purposes thereby alleviating concerns associated with the sharing of real data in some circumstances.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Synthesis of homogenous glycans in quantitative yields represents a major bottleneck to the production of molecular tools for glycoscience, such as glycan microarrays, affinity resins, and reference ...standards. Here, we describe a combined biological/enzymatic synthesis that is capable of efficiently converting microbially-derived precursor oligosaccharides into structurally uniform human-type N-glycans. Unlike starting material obtained by chemical synthesis or direct isolation from natural sources, which can be time consuming and costly to generate, our approach involves precursors derived from renewable sources including wild-type Saccharomyces cerevisiae glycoproteins and lipid-linked oligosaccharides from glycoengineered Escherichia coli. Following deglycosylation of these biosynthetic precursors, the resulting microbial oligosaccharides are subjected to a greatly simplified purification scheme followed by structural remodeling using commercially available and recombinantly produced glycosyltransferases including key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway. Using this approach, preparative quantities of hybrid and complex-type N-glycans including asymmetric multi-antennary structures were generated and subsequently used to develop a glycan microarray for high-throughput, fluorescence-based screening of glycan-binding proteins. Taken together, these results confirm our combined synthesis strategy as a new, user-friendly route for supplying chemically defined human glycans simply by combining biosynthetically-derived precursors with enzymatic remodeling.
Sound absorbing micro-perforated panels (MPPs) are being increasingly used because of their high quality in terms of hygiene, sustainability and durability. The present work investigates the ...feasibility and the performance of MPPs when used as an acoustic treatment in lecture rooms. With this purpose, three different micro-perforated steel specimens were first designed following existing predictive models and then physically manufactured through 3D additive metal printing. The specimens’ acoustic behavior was analyzed with experimental measurements in single-layer and double-layer configurations. Then, the investigation was focused on the application of double-layer MPPs to the ceiling of an existing university lecture hall to enhance speech intelligibility. Numerical simulations were carried out using a full-spectrum wave-based method: a finite-difference time-domain (FDTD) code was chosen to better handle time-dependent signals as the verbal communication. The present work proposes a workflow to explore the suitability of a specific material to speech requirements. The measured specific impedance complex values allowed to derive the input data referred to MPPs in FDTD simulations. The outcomes of the process show the influence of the acoustic treatment in terms of reverberation time (T30) and sound clarity (C50). A systematic comparison with a standard geometrical acoustic (GA) technique is reported as well.
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