An ecosystem strategy for Intel Corporation to drive adoption of its embedded solutions

Thesis (S.M. in Engineering and Management)--Massachusetts Institute of Technology, Engineering Systems Division, 2010.Cataloged from PDF version of thesis.Includes bibliographical references (p. 50).With time, successful companies and businesses grow to create a network of partners and stakeholders that work very closely with them. The very survival and growth of these companies is dependent on this ecosystem network around them. The ecosystem thrives on stakeholders mutually benefiting from each other while contributing to growth of the ecosystem itself. Every now and then business growth of such big companies with powerful ecosystems of their own is disrupted by relatively small players when incumbents have to respond. Intel, world's largest semiconductor company, has seen tremendous growth in its business since its inception. While Intel focused on continuously innovating and delivering great products for the personal computer industry, it chose not to compete in low margin embedded computing markets. Advanced RISC Machines (ARM Holdings Ltd.), a small semiconductor company during early nineties developed architecture for low power embedded computing markets that with time became the dominant architecture for mobile computing. As demand in the personal computer industry and consumer interest shifted towards portable and mobile computers, Intel delivered products for these markets. In recent years Intel, the incumbent is being threatened by ARM, the disruptor because mobile embedded platforms based on ARM architecture have encroached Intel's territory. Intel at the same time has its sight at the high growth embedded markets dominated by ARM. Today, both these players with their mature ecosystems are facing each other as they try to enter each other's territories. This Thesis analyses this classic battle for ecosystem leadership for embedded markets by Intel and ARM. Software and platform leadership is analyzed in detail and an Ecosystem strategy for Intel to drive adoption of its embedded solutions is devised in later chapters.by Prashant Paliwal.S.M.in Engineering and Managemen

Pseudomonas putida CSV86: A Candidate Genome for Genetic Bioaugmentation

Peer reviewe

Hepatitis E Virus (HEV) egress: Role of BST2 (Tetherin) and interferon induced long non- coding RNA (lncRNA) BISPR

<div><p>Background</p><p>The biology of Hepatitis E Virus (HEV), a common cause of epidemic and sporadic hepatitis, is still being explored. HEV exits liver through bile, a process which is essential for its natural transmission by feco-oral route. Though the process of this polarised HEV egress is not known in detail, HEV pORF3 and hepatocyte actin cytoskeleton have been shown to play a role.</p><p>Methods</p><p>Our transcriptome analysis in Hepatitis E virus (HEV) replicon transfected Huh7 cells at 24 and 72 hrs indicated that at 24hrs, both LncBISPR and BST2, expressed by a bidirectional promoter were highly upregulated whereas at 72 hrs, BST2 expression was comparatively reduced accompanied by normal levels of BISPR. These findings were confirmed by qPCR analysis. Co-localisation of BST2 and HEV pORF2 was confirmed in HEV transfected Huh7 by confocal microscopy. To investigate the role of BISPR/BST2 in HEV life cycle, particularly virus egress, we generated Huh7 cells with ~8kb deletion in BISPR gene using Crispr-Cas9 system. The deletion was confirmed by PCR screening, Sanger sequencing and Real time PCR. Virus egress in ΔBISPR Huh7 and Huh7 cells was compared by measuring HEV positive strand RNA copy numbers in cell lysates and culture supernatants at 24 and 72 hrs post HEV replicon transfection and further validated by western blot for HEV pORF2 capsid protein.</p><p>Results</p><p>ΔBISPR Huh7 cells showed ~8 fold increase in virus egress at 24 hrs compared to Huh7 cells. No significant difference in virus egress was observed at 72hrs. Immunohistochemistry in histologically normal liver and HEV associated acute liver failure revealed BST2 overexpression in HEV infected hepatocytes and a dominant canalicular BST2 distribution in normal liver in addition to the cytoplasmic localisation reported in literature.</p><p>Conclusions</p><p>These findings lead us to believe that BISPR and BST2 may regulate egress of HEV virions into bile <i>in vivo</i>. This system may also be used to scale up virus production <i>in vitro</i>.</p></div

Representative images showing immunohistochemical expression of BST2 (Tetherin) and HEV pORF2 <i>in vivo</i>.

<p>Expression of BST2 (Tetherin) in histologically normal liver [a-d (inset in ‘a’ represents subcapsular region)], lymph node (f) and spleen (inset f). Blood vessels in liver (b) served as positive internal control for BST2 (Tetherin) staining. Hepatocytes show cytoplasmic (c) and canalicular staining (arrows in Fig d and inset Fig d). Figs ‘e’ (liver) and ‘h’ (spleen) represent negative IHC controls with secondary antibody only. Fig ‘g’ represents negative control for pORF2 staining in histologically normal liver. Figs ‘i-t’ depict immediate serial sections of HEV associated Acute liver failure tissue biopsy (ALF) stained with Hematoxylin and Eosin (H&E; i-l), anti-HEV pORF2 IHC (m-p) and anti-BST2 (Tetherin) IHC (q-t). Similar cell populations stained with both the antibodies (Arrows in Figs l, p and t). Inset in Fig ‘s’ depicts perinuclear staining of BST2 (Tetherin) in HEV associated ALF case. Images ‘i, m, q’ were taken with 4x objective (Bar = 0.5mm); ‘a, h, g’ with 10x objective (Bar = 0.25mm); ‘b, c, d, e, f, g, j, k, n, o, r, s’ with 20x objective (Bar = 0.1mm) and ‘l, p, t, inset d and inset s’ with 40x objective (Bar = 50μm).</p

Graphical representation of RNA sequencing results.

<p>a. Bar chart representing fold changes of 15 mRNA/lncRNAs, differentially expressed in both 24 and 72 hr samples. Bars in blue and red represent fold change values of each gene at 24 and 72 hr respectively. b. Bar chart representing the major pathways observed to be enriched in Huh7 cells 24hrs post HEV transfection.c. Bar chart representing the major pathways observed to be enriched in Huh7 cells 72hrs post HEV transfection.</p

Analysis of HEV virion egress from ΔBISPR Huh7 cells.

<p>a. Bar diagram showing expression of lncBISPR and BST2 (Tetherin) in HEV replicon transfected ΔBISPR Huh7 (test) 24 hrs post transfection and untransfected Huh7 cells (control) (n = 3; error bars represent standard deviation, *p-value <0.01). GAPDH was used as a reference gene for normalization. b. Confocal images (40x objective) representing staining patterns of HEV pORF2 and BST2 (Tetherin) in untransfected and HEV transfected ΔBISPR Huh7 cells, 24hrs post transfection. Bars represent 20μm. c. Bar diagram representing percentage of HEV RNA in culture supernatants of HEV transfected ΔBISPR Huh7 and Huh7 cells at 24 hrs post transfection (n = 3; error bars represent standard deviation). d. Ponceau-S and anti-HEV pORF2 stained western blot analysis of HEV virions in the culture supernatants of HEV transfected Huh7 (lane 6) and ΔBISPR Huh7 cells (lanes 2–4) 24hrs post transfection. Capsid protein pORF2 is shown by arrow heads. Lane 5 represents the negative control sample (culture supernatant from untransfected Huh7 cells). Lane 1 represents prestained protein molecular weight marker (Puregene, United States).</p

BST2 (Tetherin) and BISPR expression in HEV transfected Huh7 cells.

<p>a. Bar chart representing BST2 (Tetherin) RNA levels in untransfected (black), wild type HEV replicon (red) and replication deficient HEV RNA (blue) transfected Huh7 cells at 12, 24 and 72 hrs post transfection. Relative expression values have been normalised with respect to GAPDH gene (n = 3; error bars represent standard deviation, *p value <0.01). b. Bar chart representing BISPR RNA levels in untransfected (black), wild type HEV replicon (red) and replication deficient HEV RNA (blue) transfected Huh7 cells at 12, 24 and 72 hrs post transfection. Relative expression values have been normalised with respect to GAPDH gene (n = 3; error bars represent standard deviation, *p value <0.01). c. Confocal images (40x objective) representing dual Immunofluorescence colocalisation patterns of HEV pORF2 and BST2 (Tetherin) in HEV transfected Huh7 cells in replicate samples (panels 1, 2: cytoplasmic and membranous, 3: perinuclear), 24hrs post transfection. Panels 4 represents staining pattern of untransfected Huh7 cells. Bars in panels 1–3 and 4 represent 20μm and 10μm respectively.</p

Design of CRISPR-Cas9 mediated lncBISPR gene deletion using dual gRNA-Cas9 and homologous recombination (HR) donor constructs.

<p>a. Genomic organisation of BST2 (Tetherin) and LncBISPR gene on human chromosome 19. b. Schematic representing the targeting sites of gRNA1 and gRNA2 in exon 2 and exon 5 of lncBISPR gene respectively. c. Schematic representing the location of primers P1 and P3 used for PCR screening of BISPR gene deletion. d. GFP positive Huh7 cells, post dual gRNA-Cas9, HR donor construct transfection and Puromycin selection (40x objective, Bar = 20μm).</p

Phylogenetic neighbor-joining tree of <i>Pseudomonas putida</i> CSV86.

<p>The tree is constructed from 16S rRNA gene sequences from 38 completely sequenced <i>Pseudomonas</i> spp. The phylogenetic analysis was performed using MEGA 5.2 and the resultant Maximum Likelihood tree shows close taxonomic relationship of <i>P. putida</i> CSV86 to <i>P. putida</i> S16.</p

New aromatics degradation pathways genes identified in <i>Pseudomonas putida</i> CSV86 by genome analysis.

<p><b>A–B</b>. Phenylpropanoid pathway genes (Contig 115, 220), <b>C–E</b>. Homogentisate pathway genes (Contig 27, 99, 177), <b>F–H</b>. Copper resistance genes (Contig 19). For details refer to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0084000#pone.0084000.s011" target="_blank">Figures S11</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0084000#pone.0084000.s012" target="_blank">S12</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0084000#pone.0084000.s013" target="_blank">S12</a>.</p