Future research to underpin successful peste des petits ruminants virus (PPRV) eradication

Peste des petits ruminants virus (PPRV) is a significant pathogen of small ruminants and is prevalent in much of Africa, the Near and Middle East and Asia. Despite the availability of an efficacious and cheap live-attenuated vaccine, the virus has continued to spread, with its range stretching from Morocco in the west to China and Mongolia in the east. Some of the world’s poorest communities rely on small ruminant farming for subsistence and the continued endemicity of PPRV is a constant threat to their livelihoods. Moreover, PPRV’s effects on the world’s population are felt broadly across many economic, agricultural and social situations. This far-reaching impact has prompted the Food and Agriculture Organization of the United Nations (FAO) and the World Organisation for Animal Health (OIE) to develop a global strategy for the eradication of this virus and its disease. PPRV is a morbillivirus and, given the experience of these organizations in eradicating the related rinderpest virus, the eradication of PPRV should be feasible. However, there are many critical areas where basic and applied virological research concerning PPRV is lacking. The purpose of this review is to highlight areas where new research could be performed in order to guide and facilitate the eradication programme. These areas include studies on disease transmission and epidemiology, the existence of wildlife reservoirs and the development of next-generation vaccines and diagnostics. With the support of the international virology community, the successful eradication of PPRV can be achieved

The mobility of Atlantic baric depressions leading to intense precipitation over Italy: a preliminary statistical analysis

International audienceThe speed of Atlantic surface depressions, occurred during the autumn and winter seasons and that lead to intense precipitation over Italy from 1951 to 2000, was investigated. Italy was divided into 5 regions as documented in previous climatological studies (based on Principal Component Analysis). Intense precipitation events were selected on the basis of in situ rain gauge data and clustered according to the region that they hit. For each intense precipitation event we tried to identify an associated surface depression and we tracked it, within a large domain covering the Mediterranean and Atlantic regions, from its formation to cyclolysis in order to estimate its speed. "Depression speeds" were estimated with 6-h resolution and clustered into slow and non-slow classes by means of a threshold, coinciding with the first quartile of speed distribution and depression centre speeds were associated with their positions. Slow speeds occurring over an area including Italy and the western Mediterranean basin showed frequencies higher than 25%, for all the Italian regions but one. The probability of obtaining by chance the observed more than 25% success rate was estimated by means of a binomial distribution. The statistical reliability of the result is confirmed for only one region. For Italy as a whole, results were confirmed at 95% confidence level. Stability of the statistical inference, with respect to errors in estimating depression speed and changes in the threshold of slow depressions, was analysed and essentially confirmed the previous results

Norovirus Polymerase Fidelity Contributes to Viral Transmission In Vivo.

Intrahost genetic diversity and replication error rates are intricately linked to RNA virus pathogenesis, with alterations in viral polymerase fidelity typically leading to attenuation during infections in vivo. We have previously shown that norovirus intrahost genetic diversity also influences viral pathogenesis using the murine norovirus model, as increasing viral mutation frequency using a mutagenic nucleoside resulted in clearance of a persistent infection in mice. Given the role of replication fidelity and genetic diversity in pathogenesis, we have now investigated whether polymerase fidelity can also impact virus transmission between susceptible hosts. We have identified a high-fidelity norovirus RNA-dependent RNA polymerase mutant (I391L) which displays delayed replication kinetics in vivo but not in cell culture. The I391L polymerase mutant also exhibited lower transmission rates between susceptible hosts than the wild-type virus and, most notably, another replication defective mutant that has wild-type levels of polymerase fidelity. These results provide the first experimental evidence that norovirus polymerase fidelity contributes to virus transmission between hosts and that maintaining diversity is important for the establishment of infection. This work supports the hypothesis that the reduced polymerase fidelity of the pandemic GII.4 human norovirus isolates may contribute to their global dominance. IMPORTANCE Virus replication fidelity and hence the intrahost genetic diversity of viral populations are known to be intricately linked to viral pathogenesis and tropism as well as to immune and antiviral escape during infection. In this study, we investigated whether changes in replication fidelity can impact the ability of a virus to transmit between susceptible hosts by the use of a mouse model for norovirus. We show that a variant encoding a high-fidelity polymerase is transmitted less efficiently between mice than the wild-type strain. This constitutes the first experimental demonstration that the polymerase fidelity of viruses can impact transmission of infection in their natural hosts. These results provide further insight into potential reasons for the global emergence of pandemic human noroviruses that display alterations in the replication fidelity of their polymerases compared to nonpandemic strains

The murine norovirus core subgenomic RNA promoter consists of a stable stem-loop that can direct accurate initiation of RNA synthesis.

UNLABELLED: All members of the Caliciviridae family of viruses produce a subgenomic RNA during infection. The subgenomic RNA typically encodes only the major and minor capsid proteins, but in murine norovirus (MNV), the subgenomic RNA also encodes the VF1 protein, which functions to suppress host innate immune responses. To date, the mechanism of norovirus subgenomic RNA synthesis has not been characterized. We have previously described the presence of an evolutionarily conserved RNA stem-loop structure on the negative-sense RNA, the complementary sequence of which codes for the viral RNA-dependent RNA polymerase (NS7). The conserved stem-loop is positioned 6 nucleotides 3' of the start site of the subgenomic RNA in all caliciviruses. We demonstrate that the conserved stem-loop is essential for MNV viability. Mutant MNV RNAs with substitutions in the stem-loop replicated poorly until they accumulated mutations that revert to restore the stem-loop sequence and/or structure. The stem-loop sequence functions in a noncoding context, as it was possible to restore the replication of an MNV mutant by introducing an additional copy of the stem-loop between the NS7- and VP1-coding regions. Finally, in vitro biochemical data suggest that the stem-loop sequence is sufficient for the initiation of viral RNA synthesis by the recombinant MNV RNA-dependent RNA polymerase, confirming that the stem-loop forms the core of the norovirus subgenomic promoter. IMPORTANCE: Noroviruses are a significant cause of viral gastroenteritis, and it is important to understand the mechanism of norovirus RNA synthesis. Here we describe the identification of an RNA stem-loop structure that functions as the core of the norovirus subgenomic RNA promoter in cells and in vitro. This work provides new insights into the molecular mechanisms of norovirus RNA synthesis and the sequences that determine the recognition of viral RNA by the RNA-dependent RNA polymerase.This is the author's accepted manuscript. The final version is available from the American Society for Microbiology at http://jvi.asm.org/content/89/2/1218.lon

An Evaluation of Bacterial Flagella Staining Procedures

Five currently used flagella stains and staining procedures have been studied and evaluated. A staining method that is simplest, most reliable, and highly reproducible, especially for beginning students and technicians has been assembled and recommended. Four important variables necessary for successful flagella staining were stressed. These variables are: growth of organisms, smear pre paration, staining procedure, and reagent stability. Two important procedures which are part of some, but not all of the flagella staining methods studied are strongly recommended. One is the practice of enclosing a smear with wax pencil and adding a known, constant volume of dye. An effective ratio of staining surface area to volume of dye used is obtained with this procedure resulting in a stan dardization that greatly diminishes overstaining and understaining reactions. The second procedure is the use of a freshly mixed dye mordant staining mixture. This procedure is the key to reproducible production of good flagella stains that have clear backgrounds and few interfering artifacts. Dye and mordant reagents may be stored separate ly for long periods of time, probably indefinitely, and still provide good, clean, reproducible flagella stains when mixed just prior to use

Functional analysis of RNA structures present at the 3' extremity of the murine norovirus genome: the variable polypyrimidine tract plays a role in viral virulence.

Interactions of host cell factors with RNA sequences and structures in the genomes of positive-strand RNA viruses play various roles in the life cycles of these viruses. Our understanding of the functional RNA elements present in norovirus genomes to date has been limited largely to in vitro analysis. However, we recently used reverse genetics to identify evolutionarily conserved RNA structures and sequences required for norovirus replication. We have now undertaken a more detailed analysis of RNA structures present at the 3′ extremity of the murine norovirus (MNV) genome. Biochemical data indicate the presence of three stable stem-loops, including two in the untranslated region, and a single-stranded polypyrimidine tract [p(Y)] of variable length between MNV isolates, within the terminal stem-loop structure. The well-characterized host cell pyrimidine binding proteins PTB and PCBP bound the 3′-untranslated region via an interaction with this variable sequence. Viruses lacking the p(Y) tract were viable both in cell culture and upon mouse infection, demonstrating that this interaction was not essential for virus replication. However, competition analysis with wild-type MNV in cell culture indicated that the loss of the p(Y) tract was associated with a fitness cost. Furthermore, a p(Y)-deleted mutant showed a reduction in virulence in the STAT1−/− mouse model, highlighting the role of RNA structures in norovirus pathogenesis. This work highlights how, like with other positive-strand RNA viruses, RNA structures present at the termini of the norovirus genome play important roles in virus replication and virulence

Design, Layout, and Testing of a Silicon Carbide-based Under Voltage Lock-out Circuit

Silicon carbide-based power devices play an increasingly important role in modern power conversion systems. Finding a means to reduce the size and complexity of these systems by even incremental amounts can have a significant impact on cost and reliability. One approach to achieving this goal is the die-level integration of gate driver circuitry with the SiC power devices. Aside from cost reductions, there are significant advantages to the integration of the gate driver circuits with the power devices. By integrating the gate driver circuitry with the power devices, the parasitic inductances traditionally seen between the gate driver and the switching devices can be significantly reduced, allowing faster switching speeds, which in turn leads to higher efficiencies, less aggressive thermal management requirements, and physically smaller passives. Collaborators from Toyota, Cree, the University of Arkansas, Oak Ridge National Labs, and Arkansas Power Electronics International have designed, fabricated, and tested a custom gate driver circuit implemented in a low-voltage SiC-based process by Cree. This gate driver implementation is the first step toward the goal of a completely integrated system. One key sub-component of this gate driver is the Under Voltage Lock-Out (UVLO) circuit, which asserts a signal whenever the supply voltage to the die falls below a set threshold and allows circuitry both on- and off-chip to take steps to prevent damage to the system. The work presented herein is the design, layout, and testing of a UVLO circuit implemented in the low-voltage silicon carbide process available from Cree. The UVLO was demonstrated to operate over a temperature range between -55 oC and 300 oC. An overview of the gate driver design, the fabrication process, and the trade-offs made during the UVLO circuit design process will be presented, as well as the integrated circuit layout workflow. A synopsis of the die testing apparatus and results will also be provided

The relationship between mathematics teachers’ content and pedagogical knowledge and their handling of student contributions: the case of Saudi trainee primary teachers

This research aimed to investigate how Saudi mathematics trainee primary teachers respond to their students’ contributions and the reasons for their actions. Also, to outline the possible influence of teachers’ knowledge, both subject matter knowledge (SMK) and pedagogical content knowledge (PCK), on their handling of students’ contributions. To achieve these goals five participants were observed for eight lessons each and interviewed. The data from three out of the five participants formed three case studies of teachers with differences in terms of their SMK and PCK. Three lesson observations for each case plus the interviews were analysed. The Knowledge Quartet framework was used to analyse the data in order to highlight the knowledge of the trainees and to direct attention towards the incidents where the teachers responded to the students’ answers. The interactions in these incidents were also analysed by the (Initiation-Response-Feedback) IRF tool to classify the teachers’ responses in relation to the students’ contributions. The findings suggest that the trainees responded to the students’ answers usually with one of two types of response. Confirmation actions in which the teachers confirmed the correctness or the fault of a given answer and questioning actions where the teachers asked further questions usually before making a decision about the answer. They reasoned their ways of accepting correct answers into two types of reasons: class-focused and individual-focused actions. Conversely, the teachers’ justification of their rejection of incorrect answers fell into three groups: protection actions, checking actions and other. Moreover, the teachers’ SMK and PCK have influenced their response to the students’ answers. Their response approach was mainly shaped by the teachers’ beliefs of how mathematics is best learnt whereas PCK influenced the quality of their responses. Also, they depended mainly on their SMK, and to some extent on their PCK, in deciding to accept or reject answers

Virtual Dashboard for Tanker Truck

Society has been very concern about the new technologies that are increasing like mushroom lately. In addition, the replacement of hardware by software has been one of the major developments in the electronics industry for many years. However, it has basically been too small to be noticed by the end user. In this new age where everything is computerized, human become more curious to learn as well as make use of new application of the virtual dashboard as it grant a significant impacts on vehicle's users. Therefore, this report present information base on the research involves in Virtual Dashboard, which is purposely design for monitoring the performance of the truck. This report features the introduction that explains about background of study, problem statement, the significant of the project, objectives and the scope of study of the project research. There is also some part of studies for supporting information or reference to support the project research. Process of identification is in short talked about in order to entire the research and describe tools of hardware and software apply to design the product. The end-result of the research is the static virtual dashboard software prototype that caters the basic needs of virtual truck. In shorts, the 2D virtual dashboard is a replacing of the collector of mechanical instruments by software-configurable display. It enabled truck driver to maintain the truck's performance while driving. Hence, the interfaces need to be simple and intuitive to use in addition to provide interaction with the users, in order to increase operation efficiency. The virtual dashboard also provides a clear visualisation from the other part of the truck, in recognizable graphics and graphs. A message window indicates the gauges' performance as well as displaying warnings when the state values approach presets limits. With this, private, secure, fast digital connection from oil tanker will be directed to the knowledge of the user