Analysis of the Genome of the Sexually Transmitted Insect Virus Helicoverpa zea Nudivirus 2

The sexually transmitted insect virus Helicoverpa zea nudivirus 2 (HzNV-2) was determined to have a circular double-stranded DNA genome of 231,621 bp coding for an estimated 113 open reading frames (ORFs). HzNV-2 is most closely related to the nudiviruses, a sister group of the insect baculoviruses. Several putative ORFs that share homology with the baculovirus core genes were identified in the viral genome. However, HzNV-2 lacks several key genetic features of baculoviruses including the late transcriptional regulation factor, LEF-1 and the palindromic hrs, which serve as origins of replication. The HzNV-2 genome was found to code for three ORFs that had significant sequence homology to cellular genes which are not generally found in viral genomes. These included a presumed juvenile hormone esterase gene, a gene coding for a putative zinc-dependent matrix metalloprotease, and a major facilitator superfamily protein gene; all of which are believed to play a role in the cellular proliferation and the tissue hypertrophy observed in the malformation of reproductive organs observed in HzNV-2 infected corn earworm moths, Helicoverpa zea

Identification of T-cell epitopes in African swine fever virus CD2v and C-type lectin proteins.

African swine fever (ASF) is an emerging disease threat for the swine industry worldwide. No ASF vaccine is available, and progress is hindered by lack of knowledge concerning the extent of ASF virus (ASFV) strain diversity and the viral antigens conferring type-specific protective immunity in pigs. Previously, we demonstrated that ASFV serotype-specific proteins CD2v (EP402R) and/or C-type lectin (EP153R) are important for protection against homologous ASF infection. Here, we identified six discrete T-cell epitope regions present on CD2v and C-type lectin using IFN-γ ELISpot assay and PBMCs from ASF immune animals, indicating cellular reactivity to these proteins in the context of ASFV infection and protective immunity. Notably, three of the epitope regions map to previously described serotype-specific signature regions of these proteins. Improved understanding of ASFV protective antigens, relevant epitopes and their diversity in nature will facilitate ASFV subunit vaccine design and development

Análise de vida à fadiga sob carregamentos não proporcionais através de modelo de dano contínuo

Dissertação (mestrado)—Universidade de Brasília, Faculdade de Tecnologia, Departamento de Engenharia Mecânica, 2016.Neste trabalho, busca-se utilizar o modelo de dano contínuo de Lemaitre como uma ferramenta de obtenção de vida em fadiga multiaxial para carregamentos proporcionais e não proporcionais. Primeiramente é apresentado o modelo matemático de Lemaitre, em que são formuladas as relações constitutivas necessárias à elaboração do modelo. O modelo de Chaboche foi escolhido para descrever a lei de evolução da tensão de endurecimento cinemático. Em seguida, é desenvolvido o modelo numérico para a solução das equações constitutivas, utilizando a lei de Chaboche com três termos e discretização implícita de Euler. Após, é feita a identificação dos parâmetros constitutivos do modelo para os aços 304 e S460N e para a liga de alumínio 6061-T6. É feita a implementação do modelo em uma rotina em linguagem FORTRAN, a qual é submetida a carregamentos uniaxiais e multiaxiais proporcionais e não proporcionais para cada material. Os dados de vida em fadiga obtidos pelo modelo de Lemaitre são, então, comparados com os dados experimentais disponíveis na literatura. É feita, em seguida, uma análise das amplitudes de tensão obtidas pelo modelo, as quais são comparadas com os dados experimentais. São obtidas, também, curvas de evolução do dano para cada carregamento e material. Os resultados obtidos mostram que o modelo de dano de Lemaitre descreve de maneira adequada o comportamento dos materiais analisados em fadiga de baixo ciclo, quando altas amplitudes de deformação são aplicadas.This project seeks to use Lemaitre’s Continuous Damage Model to obtain fatigue life estimates under multiaxial proportional and non-proportional loadings. Initially, the Lemaitre’s mathematical model is presented, by formulating the necessary constitutive relations. Chaboche’s model was chosen to describe the kinematic hardening law. Then, the numerical model necessary to solve the constitutive relations is developed, utilizing the Chaboche’s law with 3 terms and Euler’s implicit discretization. Then, the material parameters are identified for 304 and S460N steels and 6061-T6 aluminum alloy. The model is implemented in a FORTRAN routine, which is submitted to uniaxial and proportional and non-proportional multiaxial loading histories. The fatigue life data obtained from Lemaitre’s damage model is compared to experimental data. Then, a stress amplitude analysis is conducted and the numerical stresses are compared to experimental data. Damage evolution curves are also obtained for each material and loading. The results show that Lemaitre’s damage model describes adequately the behavior of the analyzed materials under low cycle fatigue, when low strain amplitudes are being applied to the specimen

Growth Kinetics and Protective Efficacy of Attenuated ASFV Strain Congo with Deletion of the EP402 Gene

African swine fever (ASF) is an emerging disease threat to the swine industry worldwide. There is no vaccine against ASF, and progress is hindered by a lack of knowledge concerning the extent of ASFV strain diversity and the viral antigens conferring type-specific protective immunity in pigs. We have previously demonstrated that homologous ASFV serotype-specific proteins CD2v (EP402R) and/or C-type lectin are required for protection against challenge with the virulent ASFV strain Congo (Genotype I, Serogroup 2), and we have identified T-cell epitopes on CD2v which may be associated with serotype-specific protection. Here, using a cell-culture adapted derivative of the ASFV strain Congo (Congo-a) with specific deletion of the EP402R gene (ΔCongoCD2v) in swine vaccination/challenge experiments, we demonstrated that deletion of the EP402R gene results in the failure of ΔCongoCD2v to induce protection against challenge with the virulent strain Congo (Congo-v). While ΔCongoCD2v growth kinetics in COS-1 cells and primary swine macrophage culture were almost identical to parental Congo-a, replication of ΔCongoCD2v in vivo was significantly reduced compared with parental Congo-a. Our data support the idea that the CD2v protein is important for the ability of homologous live-attenuated vaccines to induce protective immunity against the ASFV strain Congo challenge in vivo

Comparison of Attenuated and Virulent Strains of African Swine Fever Virus Genotype I and Serogroup 2

African swine fever (ASF) is a contagious disease of pigs caused by the ASF virus (ASFV). The main problem in the field of ASF control is the lack of vaccines. Attempts to obtain vaccines by attenuating the ASFV on cultured cell lines led to the production of attenuated viruses, some of which provided protection against infection with a homologous virus. Here we report on the biological and genomic features of the attenuated Congo-a (KK262) virus compared to its virulent homologue Congo-v (K49). Our results showed differences in in vivo replication and virulence of Congo-a. However, the attenuation of the K49 virus did not affect its ability to replicate in vitro in the primary culture of pig macrophages. Complete genome sequencing of the attenuated KK262 strain revealed an 8,8 kb deletion in the left variable region of the genome compared to the virulent homologue K49. This deletion concerned five genes of MGF360 and three genes of MGF505. In addition, three inserts in the B602L gene, genetic changes in intergenic regions and missense mutations in eight genes were detected. The data obtained contribute to a better understanding of ASFV attenuation and identification of potential virulence genes for further development of effective vaccines

Genome of Invertebrate Iridescent Virus Type 3 (Mosquito Iridescent Virus)

Iridoviruses (IVs) are classified into five genera: Iridovirus and Chloriridovirus, whose members infect invertebrates, and Ranavirus, Lymphocystivirus, and Megalocytivirus, whose members infect vertebrates. Until now, Chloriridovirus was the only IV genus for which a representative and complete genomic sequence was not available. Here, we report the genome sequence and comparative analysis of a field isolate of Invertebrate iridescent virus type 3 (IIV-3), also known as mosquito iridescent virus, currently the sole member of the genus Chloriridovirus. Approximately 20% of the 190-kbp IIV-3 genome was repetitive DNA, with DNA repeats localized in 15 apparently noncoding regions. Of the 126 predicted IIV-3 genes, 27 had homologues in all currently sequenced IVs, suggesting a genetic core for the family Iridoviridae. Fifty-two IIV-3 genes, including those encoding DNA topoisomerase II, NAD-dependent DNA ligase, SF1 helicase, IAP, and BRO protein, are present in IIV-6 (Chilo iridescent virus, prototype species of the genus Iridovirus) but not in vertebrate IVs, likely reflecting distinct evolutionary histories for vertebrate and invertebrate IVs and potentially indicative of genes that function in aspects of virus-invertebrate host interactions. Thirty-three IIV-3 genes lack homologues in other IVs. Most of these encode proteins of unknown function but also encode IIV3-053L, a protein with similarity to DNA-dependent RNA polymerase subunit 7; IIV3-044L, a putative serine/threonine protein kinase; and IIV3-080R, a protein with similarity to poxvirus MutT-like proteins. The absence of genes present in other IVs, including IIV-6; the lack of obvious colinearity with any sequenced IV; the low levels of amino acid identity of predicted proteins to IV homologues; and phylogenetic analyses of conserved proteins indicate that IIV-3 is distantly related to other IV genera

Metabolic constraints relaxed over the course of model evolution.

<p>40 simulations were carried out for 2,000 generations each. The columns represent the number of mass balances for which the constraints were relaxed.</p

Comparison of the <i>Mycoplasma gentialium model (i</i>PS189) [49] and the model presented in this paper for <i>Mycoplasma gallisepticum</i>.

<p>Comparison of the <i>Mycoplasma gentialium model (i</i>PS189) <a href="http://www.ploscompbiol.org/article/info:doi/10.1371/journal.pcbi.1003208#pcbi.1003208-Suthers1" target="_blank">[49]</a> and the model presented in this paper for <i>Mycoplasma gallisepticum</i>.</p