Reserve Price Reporting Mechanisms for Negotiation Support Systems

Information and Communication Technologies (ICT) changed our everyday business drastically. Business routines have been transformed to online activities. New theories and models were developed for the brand new online environment. For online negotiations, however, the research on new mechanisms is not enough, especially for bilateral distributive negotiations. A reserve price reporting mechanism (RPR) together with its extended version (ERPR) is proposed in this paper. The key improvement of reserve price reporting mechanisms is to let the negotiators report their reserve price to a third-party system before they actually start the negotiation. Analytical models of these mechanisms are built to prove truth revelation condition and the reduction of total social welfare comparing with traditional direct bargaining (TDB). A prototype of this RPR system is developed and a lab experiment is conducted to test the performance of the three mechanisms. The results of the experiment support that the reserve price report mechanisms proposed are more efficient than the traditional one in several dimensions

Amino acid residues in the non-structural protein 1 of porcine reproductive and respiratory syndrome virus involved in down-regulation of TNF-cx expression in vitro and attenuation in vivo

Porcine reproductive and respiratory syndrome virus (PRRSV) suppresses tumor necrosis factor-alpha (TNF-α) production at both transcriptional and post-transcriptional levels by its non-structural proteins 1α and 1β (Nsp1α and Nsp1β). To identifY the amino acid residues responsible for this activity, we generated several alanine substitution mutants of Nsp1α and Nsp1β. Examination of the mutant proteins revealed that Nsp1α residues Gly90, Asn91 , Arg97, Argl 00 and Arg124 were necessary for TNF-α promoter suppression, whereas several amino acids spanning the entire Nsp1β ~ were found to be required for this activity. Two mutant viruses, with mutations at Nsp1α Gly90 or Nsp1β residues 70-74, generated from infectious cDNA clones, exhibited attenuated viral replication in vitro and TNF-α was found to be up regulated in infected macrophages. In infected pigs, the Nsp1β mutant virus was attenuated in growth. These studies provide insights into how PRRSV evades the effector mechanisms of innate immunity dUling infection

Identification of virulence determinants of porcine reproductive and respiratory syndrome virus through construction of chimeric clones

AbstractIn order to determine virulence associated genes in porcine reproductive and respiratory syndrome virus (PRRSV), a series of chimeric viruses were generated where specific genomic regions of a highly virulent PRRSV infectious clone (FL12) were replaced with their counterparts of an attenuated vaccine strain (Prime Pac). Initial genome-wide scanning using a sow reproductive failure model indicated that non-structural (ORF 1a and 1b) and structural (ORF2-7) genomic regions appear to be sites where virulence determinants of PRRSV may reside. These results thus confirm the multigenic character of PRRSV virulence. Additional chimeras containing each individual structural ORFs (2 through 7) of Prime Pac and ORF5 of Neb-1 (parental strain of Prime Pac) within the FL12 backbone were generated and tested individually for further mapping of virulence determinants. Our results allow to conclude that NSP3–8 and ORF5 are the location of major virulence determinants, while other virulence determinants may also be contained in NSP1–3, NSP10–12 and ORF2

Characterization of a serologic marker candidate for development of a live-attenuated DIVA vaccine against porcine reproductive and respiratory syndrome virus

DIVA (differentiating infected from vaccinated animals) vaccines have proven extremely useful for control and eradication of infectious diseases in livestock. We describe here the characterization of a serologic marker epitope, so-called epitope-M201, which can be a potential target for development of a live-attenuated DIVA vaccine against porcine reproductive and respiratory syndrome virus (PRRSV). Epitope-M201 is located at the carboxyl terminus (residues 161-174) of the viral M protein. The epitope is highly immunodominant and well-conserved among type-II PRRSV isolates. Rabbit polyclonal antibodies prepared against this epitope are non-neutralizing; thus, the epitope does not seem to contribute to the protective immunity against PRRSV infection. Importantly, the immunogenicity of epitope-M201 can be disrupted through the introduction of a single amino acid mutation which does not adversely affect the viral replication. All together, our results provide an important starting point for the development of a liveattenuated DIVA vaccine against type-II PRRSV

Identification of amino acid residues important for anti-IFN activity of porcine reproductive and respiratory syndrome virus non-structural protein 1

The non-structural protein 1 (nsp1) of porcine reproductive and respiratory syndrome virus is partly responsible for inhibition of type I interferon (IFN) response by the infected host. By performing alanine-scanning mutagenesis, we have identified amino acid residues in nsp1α and nsp1β~ (the proteolytic products of nsp1) that when substituted with alanine(s) exhibited significant relief of IFNsuppression. A mutant virus (16-SA, in which residues 16-20 of nsp1β were substituted with alanines) encoding mutant nsp1β recovered from infectious cDNA clone was shown to be attenuated for growth in vitro and induced significantly higher amount of type I IFN transcripts in infected macrophages. In infected pigs, the 16-SA virus exhibited reduced growth at early times after infection but quickly regained wild type growth properties as a result of substitutions within the mutated sequences. The results indicate a strong selection pressure towards maintaining the IFN-inhibitory property of the virus for successful propagation in pigs

Amino acid residues in the non-structural protein 1 of porcine reproductive and respiratory syndrome virus involved in down-regulation of TNF-cx expression in vitro and attenuation in vivo

Porcine reproductive and respiratory syndrome virus (PRRSV) suppresses tumor necrosis factor-alpha (TNF-α) production at both transcriptional and post-transcriptional levels by its non-structural proteins 1α and 1β (Nsp1α and Nsp1β). To identifY the amino acid residues responsible for this activity, we generated several alanine substitution mutants of Nsp1α and Nsp1β. Examination of the mutant proteins revealed that Nsp1α residues Gly90, Asn91 , Arg97, Argl 00 and Arg124 were necessary for TNF-α promoter suppression, whereas several amino acids spanning the entire Nsp1β ~ were found to be required for this activity. Two mutant viruses, with mutations at Nsp1α Gly90 or Nsp1β residues 70-74, generated from infectious cDNA clones, exhibited attenuated viral replication in vitro and TNF-α was found to be up regulated in infected macrophages. In infected pigs, the Nsp1β mutant virus was attenuated in growth. These studies provide insights into how PRRSV evades the effector mechanisms of innate immunity dUling infection

Use of reverse genetics to study porcine reproductive and respiratory syndrome virus virulence

The efforts to identify important virulence determinants of the porcine reproductive and respiratory syndrome virus (PRRSV) began by comparing sequences between virulent and their avirulent derivative strains. This approach could provide an a priori clue to the candidate gene(s) that contribute to PRRSV attenuation by mutation. However, these changes may not necessarily represent the relevant acquisition of attenuation in vivo since many of these mutations may just reflect the necessary adaptation to grow in susceptible cell lines. The recent advances in reverse genetics system in PRRSV research can appropriately be used to understand the molecular basis of PRRSV virulence. The goal of this dissertation was to use a reverse genetics system to study the molecular basis of virulence and attenuation, and to identify candidate virulence determinant(s). Using Prime Pac (PP) vaccine strain as a donor and a highly virulent infectious clone (FL12) derived virus as a recipient, a series of chimeric viruses representing multiple or a single gene(s) of PP vaccine, were generated and characterized to map the virulence gene(s). These chimeric constructs were also used to generate a full-length cDNA clone (PP18) of the PP vaccine strain by serially replacing the genomic fragments of the FL12 with the corresponding regions from the PP strain. These 2 phenotypically different clone-derived viruses mimicked the in vitro (in growth kinetics in MARC-145 and PAM cells) and in vivo (in sows reproductive failure model) biological properties of their parental viruses. The results showed that the chimeric viruses comprising NSP3-8 and structural genes ORFs 2-7 were most attenuated. A fine mapping of structural gene regions by generating additional chimeric viruses with the individual structural genes and their reciprocal constructs, showed that ORF5 gene encoding major glycoprotein (GP) 5 is the most important structural gene for PRRSV virulence. In addition, ORF2 encoding GP2 and protein E, also contributed to virulence

Development of a Highly Efficient and Linear Differential CMOS Power Amplifier With Harmonic Control

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Influence of N-Linked Glycosylation of Porcine Reproductive and Respiratory Syndrome Virus GP5 on Virus Infectivity, Antigenicity, and Ability To Induce Neutralizing Antibodies

Porcine reproductive and respiratory syndrome virus (PRRSV) glycoprotein 5 (GP5) is the most abundant envelope glycoprotein and a major inducer of neutralizing antibodies in vivo. Three putative N-linked glycosylation sites (N34, N44, and N51) are located on the GP5 ectodomain, where a major neutralization epitope also exists. To determine which of these putative sites are used for glycosylation and the role of the glycan moieties in the neutralizing antibody response, we generated a panel of GP5 mutants containing amino acid substitutions at these sites. Biochemical studies with expressed wild-type (wt) and mutant proteins revealed that the mature GP5 contains high-mannose-type sugar moieties at all three sites. These mutations were subsequently incorporated into a full-length cDNA clone. Our data demonstrate that mutations involving residue N44 did not result in infectious progeny production, indicating that N44 is the most critical amino acid residue for infectivity. Viruses carrying mutations at N34, N51, and N34/51 grew to lower titers than the wt PRRSV. In serum neutralization assays, the mutant viruses exhibited enhanced sensitivity to neutralization by wt PRRSV-specific antibodies. Furthermore, inoculation of pigs with the mutant viruses induced significantly higher levels of neutralizing antibodies against the mutant as well as the wt PRRSV, suggesting that the loss of glycan residues in the ectodomain of GP5 enhances both the sensitivity of these viruses to in vitro neutralization and the immunogenicity of the nearby neutralization epitope. These results should have great significance for development of PRRSV vaccines of enhanced protective efficacy

Dynamic Feedback and Biasing for a Linear CMOS Power Amplifier with Envelope Tracking

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