scholarly journals Génétique inverse pour les virus à ARN à double brin

2009 ◽  
Vol 62 (2-4) ◽  
pp. 150
Author(s):  
H. Attoui ◽  
F. Mohd Jaafar ◽  
S. Maan ◽  
P. P.C. Mertens
Keyword(s):  
Reverse Genetics ◽  
Genetic System ◽  
Virus Genome ◽  
Dsrna Virus ◽  
Reverse Genetic System ◽  
Viral Dsrna ◽  
Viral Genomes ◽  
Virus Core ◽  
The Individual

Since the recognition by Sabin of the specific nature of reovi­ruses in 1959 and the characterisation of their genome as double-stranded ribonucleic acid (dsRNA) by Gomatos in 1960, there have been many attempts to rescue viruses by transfecting cells with viral dsRNA. These attempts were largely unsuccessful. In 1990, it was proposed by Roner and Joklik that messenger (m) RNA transcribed from orthoreovirus cores was infectious when that mRNA was transfected into cells together with rabbit reticu­locyte lysates that were pre-incubated with denatured viral dsR­NAs. However, attempts to reproduce these ‘rescue’ experiments failed in the hands of other scientists. A breakthrough came in 2007 when Kobayashi and Dermody established a plasmid-based reverse genetics system for the orthoreoviruses, which represents the first reliable, synthetic based, reverse genetic system for a dsRNA virus. The system made it possible to study the role of specified amino acids in the outer capsid protein by generating ‘designer’ mutants. A second and potentially even more signifi­cant breakthrough came in the same year when Boyce and Roy showed that mRNA transcribed from the bluetongue virus core was infectious, allowing rescue of the virus in BSR cells, a clone of BHK-21 cells. This same group extended their work to the generation of synthetic transcripts from complementary (c) DNA copies of each of the 10 genome segments cloned into plasmids, driven by the T7 polymerase. We report refinement of a T7 base transcription approach, which uses simple linear polymerase chain reaction (PCR) amplicons of the individual virus genome segments from any reovirus, for transcription of full length, fully capped mRNA transcripts. These transcripts were used to transfect BSR cells and permitted rescue of the corresponding viruses. There was no requirement for a second transfection of either capped or uncapped messages. The efficiency of the system was significantly improved, using modulators of the cells’ innate immunity; particu­larly, 2-aminopurine considerably enhanced the rescue and short­ened the time for appearance of lysis plaques. This system was successfully used for 12 segmented coltiviruses, the 12 segmented seadornaviruses and 10 segmented orbiviruses. The authors sug­gest that this simplified rescue-strategy should be applicable to any dsRNA virus, particularly the members of the 15 recognized genera of the family Reoviridae. The system is currently being tested for mono-partite dsRNA viral genomes.

scholarly journals Improved strategies for sequence-independent amplification and sequencing of viral double-stranded RNA genomes

2009 ◽  
Vol 90 (6) ◽  
pp. 1423-1432 ◽  
Author(s):  
A. C. Potgieter ◽  
N. A. Page ◽  
J. Liebenberg ◽  
I. M. Wright ◽  
O. Landt ◽  
...  
Keyword(s):  
Consensus Sequence ◽  
Virus Genome ◽  
Complete Sequence ◽  
Dsrna Virus ◽  
Viral Dsrna ◽  
Double Stranded Rna ◽  
Viral Genomes ◽  
Sequencing Technologies ◽  
Human Rotavirus ◽  
Virus Research

This paper reports significant improvements in the efficacy of sequence-independent amplification and quality of sequencing of viruses with segmented double-stranded RNA (dsRNA) genomes. We demonstrate that most remaining bottlenecks in dsRNA virus genome characterization have now been eliminated. Both the amplification and sequencing technologies used require no previous sequence knowledge of the viral dsRNA, there is no longer a need to separate genome segments or amplicons and the sequence-determined bias observed in cloning has been overcome. Combining very efficient genome amplification with pyrophosphate-based 454 (GS20/FLX) sequencing enabled sequencing of complete segmented dsRNA genomes and accelerated the sequence analysis of the amplified viral genomes. We report the complete consensus sequence of seven viruses from four different dsRNA virus groups, which include the first complete sequence of the genome of equine encephalosis virus (EEV), the first complete sequence of an African horsesickness virus (AHSV) genome determined directly from a blood sample and a complete human rotavirus genome determined from faeces. We also present the first comparison between the complete consensus sequence of a virulent and an attenuated strain of AHSV1. Ultra-deep sequencing (>400-fold coverage) of the AHSV1 reference and attenuated strains revealed different ratios of reassortants in the reference strain and allowed quasispecies detection in the plaque-purified attenuated strain of AHSV1. This approach amounts to a paradigm shift in dsRNA virus research, since it is sensitive and specific enough for comprehensive investigations of the evolution and genetic diversity in dsRNA virus populations.

scholarly journals Intra-Colonial Viral Infections in Western Honey Bees (Apis mellifera)

2021 ◽  
Vol 9 (5) ◽  
pp. 1087
Author(s):  
Loreley Castelli ◽  
María Laura Genchi García ◽  
Anne Dalmon ◽  
Daniela Arredondo ◽  
Karina Antúnez ◽  
...  
Keyword(s):  
Apis Mellifera ◽  
Honey Bee ◽  
Viral Infections ◽  
Sampling Period ◽  
Virus Genome ◽  
The Individual ◽  
Bee Virus ◽  
Insight Into ◽  
Bee Viruses

RNA viruses play a significant role in the current high losses of pollinators. Although many studies have focused on the epidemiology of western honey bee (Apis mellifera) viruses at the colony level, the dynamics of virus infection within colonies remains poorly explored. In this study, the two main variants of the ubiquitous honey bee virus DWV as well as three major honey bee viruses (SBV, ABPV and BQCV) were analyzed from Varroa-destructor-parasitized pupae. More precisely, RT-qPCR was used to quantify and compare virus genome copies across honey bee pupae at the individual and subfamily levels (i.e., patrilines, sharing the same mother queen but with different drones as fathers). Additionally, virus genome copies were compared in cells parasitized by reproducing and non-reproducing mite foundresses to assess the role of this vector. Only DWV was detected in the samples, and the two variants of this virus significantly differed when comparing the sampling period, colonies and patrilines. Moreover, DWV-A and DWV-B exhibited different infection patterns, reflecting contrasting dynamics. Altogether, these results provide new insight into honey bee diseases and stress the need for more studies about the mechanisms of intra-colonial disease variation in social insects.

scholarly journals Tet-Inducible Production of Infectious Zika Virus from the Full-Length cDNA Clones of African- and Asian-Lineage Strains

Viruses ◽  
2018 ◽  
Vol 10 (12) ◽  
pp. 700 ◽  
Author(s):  
Lizhou Zhang ◽  
Wei Ji ◽  
Shuang Lyu ◽  
Luhua Qiao ◽  
Guangxiang Luo
Keyword(s):  
Zika Virus ◽  
Reverse Genetics ◽  
Genetic System ◽  
Full Length ◽  
Reverse Genetic System ◽  
Cdna Clones ◽  
Viral Pathogen ◽  
Stable Plasmid ◽  
Asian Lineage ◽  
Delta Virus

Zika virus (ZIKV) is a mosquito-borne flavivirus that has emerged as an important human viral pathogen, causing congenital malformation including microcephaly among infants born to mothers infected with the virus during pregnancy. Phylogenetic analysis suggested that ZIKV can be classified into African and Asian lineages. In this study, we have developed a stable plasmid-based reverse genetic system for robust production of both ZIKV prototype African-lineage MR766 and clinical Asian-lineage FSS13025 strains using a tetracycline (Tet)-controlled gene expression vector. Transcription of the full-length ZIKV RNA is under the control of the Tet-responsive Ptight promoter at the 5′ end and an antigenomic ribozyme of hepatitis delta virus at the 3′ end. The transcription of infectious ZIKV RNA genome was efficiently induced by doxycycline. This novel ZIKV reverse genetics system will be valuable for the study of molecular viral pathogenesis of ZIKV and the development of new vaccines against ZIKV infection.

scholarly journals ZWA: Viral genome assembly and characterization hindrances from virus-host chimeric reads; a refining approach

2021 ◽  
Vol 17 (8) ◽  
pp. e1009304
Author(s):  
Nikolas Dovrolis ◽  
Katerina Kassela ◽  
Konstantinos Konstantinidis ◽  
Adamantia Kouvela ◽  
Stavroula Veletza ◽  
...  
Keyword(s):  
Genome Assembly ◽  
Genetic Material ◽  
Emerging Infectious Diseases ◽  
Virus Genome ◽  
Viral Metagenomics ◽  
Substantial Impact ◽  
Viral Genomes ◽  
Assembly Pipeline ◽  
Host Genetic

Viral metagenomics, also known as virome studies, have yielded an unprecedented number of novel sequences, essential in recognizing and characterizing the etiological agent and the origin of emerging infectious diseases. Several tools and pipelines have been developed, to date, for the identification and assembly of viral genomes. Assembly pipelines often result in viral genomes contaminated with host genetic material, some of which are currently deposited into public databases. In the current report, we present a group of deposited sequences that encompass ribosomal RNA (rRNA) contamination. We highlight the detrimental role of chimeric next generation sequencing reads, between host rRNA sequences and viral sequences, in virus genome assembly and we present the hindrances these reads may pose to current methodologies. We have further developed a refining pipeline, the Zero Waste Algorithm (ZWA) that assists in the assembly of low abundance viral genomes. ZWA performs context-depended trimming of chimeric reads, precisely removing their rRNA moiety. These, otherwise discarded, reads were fed to the assembly pipeline and assisted in the construction of larger and cleaner contigs making a substantial impact on current assembly methodologies. ZWA pipeline may significantly enhance virus genome assembly from low abundance samples and virus metagenomics approaches in which a small number of reads determine genome quality and integrity.

scholarly journals Viral Pathogenesis, Recombinant Vaccines, and Oncolytic Virotherapy: Applications of the Canine Distemper Virus Reverse Genetics System

Viruses ◽  
2020 ◽  
Vol 12 (3) ◽  
pp. 339 ◽  
Author(s):  
Jianjun Zhao ◽  
Yanrong Ren ◽  
Jie Chen ◽  
Jiasan Zheng ◽  
Dongbo Sun
Keyword(s):  
Canine Distemper Virus ◽  
Reverse Genetics ◽  
Tumor Regression ◽  
Genetic System ◽  
Viral Pathogenesis ◽  
Oncolytic Viruses ◽  
Reverse Genetic System ◽  
Canine Distemper ◽  
Host Tropism ◽  
Signaling Lymphocytic Activation Molecule

Canine distemper virus (CDV) is a highly contagious pathogen transmissible to a broad range of terrestrial and aquatic carnivores. Despite the availability of attenuated vaccines against CDV, the virus remains responsible for outbreaks of canine distemper (CD) with significant morbidity and mortality in domesticated and wild carnivores worldwide. CDV uses the signaling lymphocytic activation molecule (SLAM, or CD150) and nectin-4 (PVRL4) as entry receptors, well-known tumor-associated markers for several lymphadenomas and adenocarcinomas, which are also responsible for the lysis of tumor cells and apparent tumor regression. Thus, CDV vaccine strains have emerged as a promising platform of oncolytic viruses for use in animal cancer therapy. Recent advances have revealed that use of the CDV reverse genetic system (RGS) has helped increase the understanding of viral pathogenesis and explore the development of recombinant CDV vaccines. In addition, genetic engineering of CDV based on RGS approaches also has the potential of enhancing oncolytic activity and selectively targeting tumors. Here, we reviewed the host tropism and pathogenesis of CDV, and current development of recombinant CDV-based vaccines as well as their use as oncolytic viruses against cancers.

School Success and School Engagement of Immigrant Children and Adolescents

2013 ◽  
Vol 18 (2) ◽  
pp. 126-135 ◽  
Author(s):  
Frosso Motti-Stefanidi ◽  
Ann S. Masten
Keyword(s):  
Academic Achievement ◽  
Academic Success ◽  
Children And Adolescents ◽  
School Engagement ◽  
School Success ◽  
Immigrant Children ◽  
Integrative Model ◽  
High Stakes ◽  
The Individual

Academic achievement in immigrant children and adolescents is an indicator of current and future adaptive success. Since the future of immigrant youths is inextricably linked to that of the receiving society, the success of their trajectory through school becomes a high stakes issue both for the individual and society. The present article focuses on school success in immigrant children and adolescents, and the role of school engagement in accounting for individual and group differences in academic achievement from the perspective of a multilevel integrative model of immigrant youths’ adaptation ( Motti-Stefanidi, Berry, Chryssochoou, Sam, & Phinney, 2012 ). Drawing on this conceptual framework, school success is examined in developmental and acculturative context, taking into account multiple levels of analysis. Findings suggest that for both immigrant and nonimmigrant youths the relationship between school engagement and school success is bidirectional, each influencing over time the other. Evidence regarding potential moderating and mediating roles of school engagement for the academic success of immigrant youths also is evaluated.

Role of Transcriptional Read-Through in PRE Activity in Drosophila melanogaster

Acta Naturae ◽  
2016 ◽  
Vol 8 (2) ◽  
pp. 79-86 ◽  
Author(s):  
P. V. Elizar’ev ◽  
D. V. Lomaev ◽  
D. A. Chetverina ◽  
P. G. Georgiev ◽  
M. M. Erokhin
Keyword(s):  
Dna Sequences ◽  
Cell Types ◽  
Transcription Terminator ◽  
Response Elements ◽  
Polycomb Response Elements ◽  
The Individual ◽  
Multicellular Organisms ◽  
Different Cell Types ◽  
Main Factor

Maintenance of the individual patterns of gene expression in different cell types is required for the differentiation and development of multicellular organisms. Expression of many genes is controlled by Polycomb (PcG) and Trithorax (TrxG) group proteins that act through association with chromatin. PcG/TrxG are assembled on the DNA sequences termed PREs (Polycomb Response Elements), the activity of which can be modulated and switched from repression to activation. In this study, we analyzed the influence of transcriptional read-through on PRE activity switch mediated by the yeast activator GAL4. We show that a transcription terminator inserted between the promoter and PRE doesnt prevent switching of PRE activity from repression to activation. We demonstrate that, independently of PRE orientation, high levels of transcription fail to dislodge PcG/TrxG proteins from PRE in the absence of a terminator. Thus, transcription is not the main factor required for PRE activity switch.

УКРАЇНОЗНАВЧИЙ ОСВІТНІЙ ЧИННИК В УМОВАХ СУЧАСНОЇ УКРАЇНСЬКОЇ МАСОВОЇ КУЛЬТУРИ

2019 ◽  
pp. 22-29
Author(s):  
Н. В. Фрадкіна
Keyword(s):  
Mass Culture ◽  
Personal Development ◽  
Negative Impact ◽  
Educational Process ◽  
Point Of View ◽  
Character Trait ◽  
Main Task ◽  
Leading Role ◽  
The Individual

The purpose and tasks of the work are to analyze the contemporary Ukrainian mass culture in terms of its value and humanistic components, as well as the importance of cultural studies and Ukrainian studies in educational disciplines for the formation of a holistic worldview of modern youth.Analysis of research and publications. Scientists repeatedly turned to the problems of the role of spirituality in the formation of society and its culture. This problem is highlighted in the publications by O. Losev, V. Lytvyn, D. Likhachev, S. Avierintsev, M. Zakovych, I. Stepanenko and E. Kostyshyn.Experts see the main negative impact of mass culture on the quality approach, which determines mass culture through the market, because mass culture, from our point of view, is everything that is sold and used in mass demand.One of the most interesting studies on this issue was the work by the representatives of Frankfurt School M. Horkheimer and T. Adorno «Dialectics of Enlightenment» (1947), devoted to a detailed analysis of mass culture. Propaganda at all socio-cultural levels in the form is similar in both totalitarian and democratic countries. It is connected, according to the authors, with the direction of European enlightenment. The tendency to unify people is a manifestation of the influence of mass culture, from cinema to pop. Mass culture is a phenomenon whose existence is associated with commerce (accumulation in any form – this is the main feature of education), in general, the fact that it exists in this form is related to the direction of the history of civilization.Modern mass culture, with its externally attractive and easily assimilated ideas and symbols, appealing to the trends of modern fashion, becomes a standard of prestigious consumption, does not require intense reflection, allows you to relax, distract, not teach, but entertains, preaches hedonism as the main spiritual value. And as a consequence, there are socio-cultural risks: an active rejection of other people, which leads to the formation of indifference; cruelty as a character trait; increase of violent and mercenary crime; increase in the number of alcohol and drug addicts; anti-patriotism; indifference to the values of the family and as a result of social orphanhood and prostitution.Conclusions, perspectives of research. Thus, we can conclude that modern Ukrainian education is predominantly formed by the values of mass culture. Namely, according to the «Dialectic» by Horkheimer and Adorno, «semi-enlightenment becomes an objective spirit» of our modern society.It is concluded that only high-quality education can create the opposite of the onset of mass culture and the destruction of spirituality in our society. It is proved that only by realizing the importance of cultivating disciplines in the educational process and the spiritual upbringing of the nation, through educational reforms, humanitarian knowledge will gradually return to student audiences.Formation of youth occurs under the influence of social environment, culture, education and self-education. The optimal combination of these factors determines both the process of socialization itself and how successful it will be. In this context, one can see the leading role of education and upbringing. It turns out that the main task of modern education is to spread its influence on the development of spiritual culture of the individual, which eventually becomes a solid foundation for the formation of the individual. Such a subject requires both philosophical and humanitarian approaches in further integrated interdisciplinary research, since the availability of such research will provide the theoretical foundation for truly modern educational and personal development.

MEDIA SOCIALIZATION OF PERSONALITY IN THE CONTEXT OF INMUTATION IN THE MASS MEDIA

2021 ◽  
pp. 104-109
Author(s):  
Chernysh O.O.
Keyword(s):  
Mass Media ◽  
Negative Impact ◽  
Effective Interaction ◽  
Individual Characteristics ◽  
Media Culture ◽  
Active Growth ◽  
Media Space ◽  
The Media ◽  
The Individual

The urgency of the researched problem is connected with the growing role of mass media in modern conditions leads to change of values and transformation of identity of the person. The active growth of the role of the media, their influence on the formation and development of personality leads to the concept of “media socialization” and immutation in the media. The aim of the study is to outline the possibilities of the process of media socialization in the context of immutation in the media. The methods of our research are: analysis of pedagogical, psychological, literature, synthesis, comparison, generalization. The article analyzes the views of domestic and foreign scientists on the problem of immutation in the media and the transformation of the information space. In the context of the mass nature of the immutation of society, the concept of “media socialization” becomes relevant, which is the basis for reducing the negative impact of the media on the individual.The author identifies the lack of a thorough study of the concept of “media socialization” in modern scientific thought. Thus, media socialization is associated with the transformation of traditional means of socialization, and is to assimilate and reproduce the social experience of mankind with the help of new media.The article analyzes the essence of the concepts “media space”, “mass media” and “immutation”. The influence of mass media on the formation and development of the modern personality is described in detail.The study concluded that it is necessary to form a media culture of the individual, to establish safe and effective interaction of young people with the modern media system, the formation of media awareness, media literacy and media competence in accordance with age and individual characteristics for successful media socialization. The role of state bodies in solving the problem of media socialization of the individual was also determined. It is determined that the process of formation of media culture in youth should take place at the level of traditional institutions of socialization of the individual.The author sees the prospect of further research in a detailed analysis and study of the potential of educational institutions as an institution and a means of counteracting the mass nature of the immutation of society.Key words: immutation, media socialization, mass media, media space, information.

The Role of Threat and Economic Uncertainty in Support for Scottish Independence

2014 ◽  
Vol 23 (1) ◽  
pp. 103-124 ◽  
Author(s):  
Daniel Kopasker
Keyword(s):  
Economic Consequences ◽  
Perceived Threat ◽  
Constitutional Change ◽  
Lack Of Information ◽  
Economic Expectations ◽  
Scottish Independence ◽  
The Individual ◽  
The Impact ◽  
Levels Of Support

Existing research has consistently shown that perceptions of the potential economic consequences of Scottish independence are vital to levels of support for constitutional change. This paper attempts to investigate the mechanism by which expectations of the economic consequences of independence are formed. A hypothesised causal micro-level mechanism is tested that relates constitutional preferences to the existing skill investments of the individual. Evidence is presented that larger skill investments are associated with a greater likelihood of perceiving economic threats from independence. Additionally, greater perceived threat results in lower support for independence. The impact of uncertainty on both positive and negative economic expectations is also examined. While uncertainty has little effect on negative expectations, it significantly reduces the likelihood of those with positive expectations supporting independence. Overall, it appears that a general economy-wide threat is most significant, and it is conjectured that this stems a lack of information on macroeconomic governance credentials.

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