Resynthesis Algorithm for Avoiding Undetectable Faults based on Design-For-Manufacturability Guidelines

Due to shrinking feature sizes, the number of defects in electronic chips has risen. These defects can be modeled as logic faults. Tests are applied to check whether or not a fault is present. Faults are not always present, but always need to be tested for. If a test for a fault does not exist, the fault is called undetectable. Undetectable faults leave uncovered defect sites in the circuit. It has been observed that undetectable faults cluster in sub-circuits of benchmark circuits. This implies that certain sub-circuits are uncovered or less covered than others by a test set for all the modeled faults. Design-For-Manufacturability (DFM) guidelines were created which if followed during manufacturing, can help avoid potential defects. DFM guidelines are not followed if they conflict with other constraints such as area or performance. This implies that defects are not avoided even if the possibility of their occurrence can be predicted. This research involves resynthesizing sub-circuits by using an algorithm in order to reduce the number of undetectable internal faults in them and break the fault clusters. It was observed that by replacing one instance in one cell type in every iteration, the number of undetectable internal faults were successfully reduced. It was also observed that in slightly larger circuits, the number of faults increased at the beginning of every new cycle due to an optimization step in the tool. It can thus be concluded that it is possible to break down the undetectable fault clusters using resynthesis

Integration and expression of Bluetongue VP2 gene in somatic embryos of peanut through particle bombardment method

After pre-culture and treatment of osmosis, zygotic embryos of peanut (Arachis hypogaea L.) were transformed via particle bombardment with a plasmid containing a Bluetongue VP2 gene (BTVP2) comprising neutralizing epitopes. Selection for Kanamycin resistant calluses and somatic embryos was initiated at 12th day post-bombardment on medium containing 25 mg/L Kanamycin. Under continuous selection, 12.38 Kanamycin resistant plantlets were regenerated from bombarded somatic embryos. The presence and integration of BTVP2 DNA in regenerated Kanamycin resistant plants were confirmed by southern hybridization assay using non-radioactive Digoxiginin BTVP2 probe. β-Glucuronidase (GUS) enzyme activity was detected in transgenic somatic embryos but not from control, non-transformed embryos. The expression of the BTVP2 protein was confirmed through RT-PCR (reverse transcription polymerase chain reaction) using the RNA isolated from the transgenic callus employing BTVP2-specific primers. The production of transgenic peanut was mainly focused on evaluating a newly improved somatic embryogenesis regeneration system as well as the gene transfer method and to produce the Bluetongue outer coat protein that comprises the neutralizing epitopes. © 2005 Elsevier Ltd. All rights reserved

Investigation of greenhouse gas reduction strategies by industries : an enterprise systems architecting approach

Thesis (S.M. in Engineering and Management)--Massachusetts Institute of Technology, Engineering Systems Division, 2012.Cataloged from PDF version of thesis.Includes bibliographical references (p. 85-89).This thesis explores an enterprise systems architecting approach to investigate the greenhouse gas reduction strategies followed by industries, especially for automotive industry and Information Technology industry. The strategic dimensions of greenhouse gas reduction aspects-drivers, actions and challenges-faced by industries are identified and a survey was circulated among the senior and mid-level managers of both industries. The survey results are compiled and analyzed to understand the leading drivers, actions and challenges in addition to the ranking of the eight views of enterprise architecting. The results are then used to identify gaps between the current status and the envisioned future state of the companies, based on the survey results, internal assessments and prevailing state of the art greenhouse gas reduction strategies. Several candidate architectures are developed based on the identified gaps for both industries. An alignment matrix for all the eight views with the candidate architectures is also developed. Generic frameworks to evaluate candidate architectures using 'ilities' and weighting factors are discussed. Greenhouse gas profiles of both the industries are compared, and future research scope to extend this thesis is presented.by Kumaresh Babu Tanthullu Athmaram.S.M.in Engineering and Managemen

Characterization of Protection Afforded by a Bivalent Virus-Like Particle Vaccine against Bluetongue Virus Serotypes 1 and 4 in Sheep

BACKGROUND: Bluetongue virus (BTV) is an economically important, arthropod borne, emerging pathogen in Europe, causing disease mainly in sheep and cattle. Routine vaccination for bluetongue would require the ability to distinguish between vaccinated and infected individuals (DIVA). Current vaccines are effective but are not DIVA. Virus-like particles (VLPs) are highly immunogenic structural mimics of virus particles, that only contain a subset of the proteins present in a natural infection. VLPs therefore offer the potential for the development of DIVA compatible bluetongue vaccines. METHODOLOGY/PRINCIPAL FINDINGS: Merino sheep were vaccinated with either monovalent BTV-1 VLPs or a bivalent mixture of BTV-1 VLPs and BTV-4 VLPs, and challenged with virulent BTV-1 or BTV-4. Animals were monitored for clinical signs, antibody responses, and viral RNA. 19/20 animals vaccinated with BTV-1 VLPs either alone or in combination with BTV-4 VLPs developed neutralizing antibodies to BTV-1, and group specific antibodies to BTV VP7. The one animal that showed no detectable neutralizing antibodies, or group specific antibodies, had detectable viral RNA following challenge but did not display any clinical signs on challenge with virulent BTV-1. In contrast, all control animals' demonstrated classical clinical signs for bluetongue on challenge with the same virus. Six animals were vaccinated with bivalent vaccine and challenged with virulent BTV-4, two of these animals had detectable viral levels of viral RNA, and one of these showed clinical signs consistent with BTV infection and died. CONCLUSIONS: There is good evidence that BTV-1 VLPs delivered as monovalent or bivalent immunogen protect from bluetongue disease on challenge with virulent BTV-1. However, it is possible that there is some interference in protective response for BTV-4 in the bivalent BTV-1 and BTV-4 VLP vaccine. This raises the question of whether all combinations of bivalent BTV vaccines are possible, or if immunodominance of particular serotypes could interfere with vaccine efficacy

Yeast expressed recombinant Hemagglutinin protein of Novel H1N1 elicits neutralising antibodies in rabbits and mice

Currently available vaccines for the pandemic Influenza A (H1N1) 2009 produced in chicken eggs have serious impediments viz limited availability, risk of allergic reactions and the possible selection of sub-populations differing from the naturally occurring virus, whereas the cell culture derived vaccines are time consuming and may not meet the demands of rapid global vaccination required to combat the present/future pandemic. Hemagglutinin (HA) based subunit vaccine for H1N1 requires the HA protein in glycosylated form, which is impossible with the commonly used bacterial expression platform. Additionally, bacterial derived protein requires extensive purification and refolding steps for vaccine applications. For these reasons an alternative heterologous system for rapid, easy and economical production of Hemagglutinin protein in its glycosylated form is required. The HA gene of novel H1N1 A/California/04/2009 was engineered for expression in Pichia pastoris as a soluble secreted protein. The full length HA- synthetic gene having α-secretory tag was integrated into P. pastoris genome through homologous recombination. The resultant Pichia clones having multiple copy integrants of the transgene expressed full length HA protein in the culture supernatant. The Recombinant yeast derived H1N1 HA protein elicited neutralising antibodies both in mice and rabbits. The sera from immunised animals also exhibited Hemagglutination Inhibition (HI) activity. Considering the safety, reliability and also economic potential of Pichia expression platform, our preliminary data indicates the feasibility of using this system as an alternative for large-scale production of recombinant influenza HA protein in the face of influenza pandemic threat

A Novel Role for IκBζ in the Regulation of IFNγ Production

IκBζ is a novel member of the IκB family of NFκB regulators, which modulates NFκB activity in the nucleus, rather than controlling its nuclear translocation. IκBζ is specifically induced by IL-1β and several TLR ligands and positively regulates NFκB-mediated transcription of genes such as IL-6 and NGAL as an NFκB binding co-factor. We recently reported that the IL-1 family cytokines, IL-1β and IL-18, strongly synergize with TNFα for IFNγ production in KG-1 cells, whereas the same cytokines alone have minimal effects on IFNγ production. Given the striking similarities between the IL-1R and IL-18R signaling pathways we hypothesized that a common signaling event or gene product downstream of these receptors is responsible for the observed synergy. We investigated IκBζ protein expression in KG-1 cells upon stimulation with IL-1β, IL-18 and TNFα. Our results demonstrated that IL-18, as well as IL-1β, induced moderate IκBζ expression in KG-1 cells. However, TNFα synergized with IL-1β and IL-18, whereas by itself it had a minimal effect on IκBζ expression. NFκB inhibition resulted in decreased IL-1β/IL-18/TNFα-stimulated IFNγ release. Moreover, silencing of IκBζ expression led to a specific decrease in IFNγ production. Overall, our data suggests that IκBζ positively regulates NFκB-mediated IFNγ production in KG-1 cells

Multiple large foreign protein expression by a single recombinant baculovirus: a system for production of multivalent vaccines.

Baculovirus expression system offers the advantage of expression of several large proteins simultaneously by a single recombinant virus. To date, expression of multiple large (>100kDa) proteins has been hampered by the need to generate large constructs and repeat use of homologous sequence and promoter. The development of multi-loci baculovirus expression system overcomes these issues by enabling the recombination of large foreign sequences into different regions of the genome. In this paper, we have examined the co-expression of African horse sickness virus (AHSV) VP2 proteins from multiple serotypes in a single recombinant baculovirus. To this end, recombinant baculoviruses expressing multiple AHSV VP2 proteins were generated and it was found that up to six different AHSV serotypes (serotype 1, 3, 4, 5, 7 and 8) VP2 proteins (∼120kDa) could be expressed simultaneously from different loci of baculovirus genome. The expression of VP2 of one serotype was not significantly hindered by the presence of other serotypes, although there were slight differences in expression level between different serotypes. The expression of VP2 of further serotypes from additional loci resulted in a lesser expression level of VP2 proteins. Based on these findings, three additional recombinant baculoviruses encompassing all nine AHSV serotypes were constructed (serotypes 1, 7, 8 or serotypes 2, 4, 5 or serotypes 3, 6, 9) and each of the triple recombinant viruses exhibited similar expression level of each VP2. This system allows for the expression of a number of large proteins that has the potential to be exploited for multivalent vaccines production

Sonographic Consensual Pupillary Reflex

Patients suffering from severe orbital trauma are at risk for numerous complications, including orbital compartment syndromes. This can result in an afferent pupillary defect, which must be evaluated for on physical examination. Unfortunately, these at-risk patients are often challengingto examine properly due to surrounding edema. Point-of-care ultrasonography can be used as an adjunct to the standard examination in this situation. [West J Emerg Med. 2012;13(6):524

Identification of two new genes from drought tolerant peanut up-regulated in response to drought

Using differential display of mRNA transcripts, we obtained nine partial cDNA clones that were up-regulated and down-regulated by exposure to drought. Two full length cDNAs of 825 bp and 700 bp, designated as Arachis hypogaea serine rich protein (AhSrp) and Arachis hypogaea leucine rich protein (AhLrp) were obtained using RLM-RACE. Based on the cDNA sequence, the transcriptional start site of AhSrp and AhLrp was assigned. AhSrp is predicted to code for a protein of 233 amino acids containing a signal peptide, three potential transmembrane domains and a sumolyation motif. AhLrp is predicted to code for a protein of 206 amino acids containing a signal peptide. The expression of partial cDNA and full-length cDNA were confirmed by RNA dot blot analysis. Expression of AhSrp was observed between 6 and 10 days of stress and prominent expression of AhLrp was visualized at 10 days of stress by semi quanititative PCR. Our studies have resulted in the identification of two new genes that may play a role in plant response to drought stress in peanut. © 2007 Springer Science+Business Media B.V