Leaders\u27 Accounts: A Study of Transnational NGOs Leadership Views on Accountability

The increased involvement of non-state actors such as nongovernmental organizations (NGOs) in The increased involvement of non-state actors such as nongovernmental organizations (NGOs) in shaping various national and global policy processes raises serious questions about the accountability, authority, and legitimacy of these important yet unelected actors. Although there is no consensus on how to define accountability, broadly the concept refers to the idea that some hold others responsible for their actions according to a set of standards defining proper behavior. However, deciding to whom and to what standards NGOs are accountable and how they are (or should be) held accountable remain central and open questions in accountability debates. This study relies on both qualitative and quantitative methods, introducing the use of qualitative pattern analysis to understand not only what leaders of TNGOs have to say about accountability, but also how they describe their accountability experiences. It uses the TNGO initiative datasets, which include in-depth interview data for 152 CEOs of U.S.- registered TNGOs, as well as secondary data collected for each of the organizations in the sample. Drawing from the nonprofit, management, and international relations scholarship, I explore the three central questions of accountability debates: What is accountability? Accountable to whom? And how? I argue that once one addresses each of these accountability questions, taking an actor-centered perspective, it is possible to understand how accountability practices vary across transnational NGOs. I propose the concept of accountability dissonance to explain the disconnect between the three main questions of accountability. Through the empirical chapters, I show that TNGO leaders expressed often complex views about their accountability experiences, yet they generally struggled to communicate their practices. I stress the need to pay attention to the substantive meaning of accountability messaging and to the connections among definitions, audiences, and responses. The phenomenon of accountability dissonance persists because leaders express their accountability using tools and processes that are mismatched to their definitions and audiences. Furthermore, the data suggest that the types of organizations TNGO leaders manage matter in very different ways, contingent on the aspect of accountability being considered. To reach a holistic understanding, I propose an overarching framework using the analogy of the puzzle to highlight the need for a more integrated approach to TNGO accountability, one hinging on the ability to target the communication of accountability performance to specific audiences

JPEG steganography with particle swarm optimization accelerated by AVX

Digital steganography aims at hiding secret messages in digital data transmitted over insecure channels. The JPEG format is prevalent in digital communication, and images are often used as cover objects in digital steganography. Optimization methods can improve the properties of images with embedded secret but introduce additional computational complexity to their processing. AVX instructions available in modern CPUs are, in this work, used to accelerate data parallel operations that are part of image steganography with advanced optimizations.Web of Science328art. no. e544

Expression of Bovine Interleukin-1β in a Bovine Herpesvirus-1 Vector:In VitroAnalysis

AbstractIn order to evaluate whether bovine herpesvirus-1 (BHV-1) could be used as a live viral vector for the expression of cytokines, we constructed a recombinant BHV-1 expressing bovine interleukin-1β (boIL-1β). The boIL-1β coding sequence, corresponding to the cleaved mature product, was fused with the BHV-1 glycoprotein C (gC) signal peptide sequence; the resultant gC-boIL-1β fusion gene was recombined into the gC locus of the BHV-1 genome. Southern blot analysis confirmed the proper genomic configuration of the recombinant virus. Results from transcript analysis showed that boIL-1β was expressed in infected cells with kinetics similar to that of gC. Indirect immunofluorescence and immunoprecipitation assays showed that the recombinant protein was produced in both cell-associated and secreted forms. Western blot analysis detected a 19.3-kDa protein. Further analysis, using an IL-1β bioassay demonstrated that both the cellular and secreted forms of recombinant boIL-1β possessed biological activity. The expression of the boIL-1β protein did not affect thein vitrogrowth efficiency of the virus, which exhibited similar growth kinetics to that of a simple gC deletion mutant. The results from this study demonstrate that BHV-1 can be used to express a functional cytokine, thereby establishing the basis to further study recombinant BHV-1 expressing cytokines as an alternative means to attenuate the virus and also as a potentialin situcytokine delivery system to modulate immune responses against BHV-1 and other cattle pathogens

The site-2 protease

AbstractThe site-2 protease (S2P) is an unusually-hydrophobic integral membrane protease. It cleaves its substrates, which are membrane-bound transcription factors, within membrane-spanning helices. Although structural information for S2P from animals is lacking, the available data suggest that cleavage may occur at or within the lipid bilayer. In mammalian cells, S2P is essential owing to its activation of the sterol regulatory element binding proteins (SREBPs); in the absence of exogenous lipid, cells lacking S2P cannot survive. S2P is also important in the endoplasmic reticulum (ER) stress response, activating several different membrane-bound transcription factors. Human patients harboring reduction-of-function mutations in S2P exhibit an array of pathologies ranging from skin defects to neurological abnormalities. Surprisingly, Drosophila melanogaster lacking S2P are viable and fertile. This article is part of a Special Issue entitled: Intramembrane Proteases

Adaptation to ER Stress Is Mediated by Differential Stabilities of Pro-Survival and Pro-Apoptotic mRNAs and Proteins

The accumulation of unfolded proteins in the endoplasmic reticulum (ER) activates a signaling cascade known as the unfolded protein response (UPR). Although activation of the UPR is well described, there is little sense of how the response, which initiates both apoptotic and adaptive pathways, can selectively allow for adaptation. Here we describe the reconstitution of an adaptive ER stress response in a cell culture system. Monitoring the activation and maintenance of representative UPR gene expression pathways that facilitate either adaptation or apoptosis, we demonstrate that mild ER stress activates all UPR sensors. However, survival is favored during mild stress as a consequence of the intrinsic instabilities of mRNAs and proteins that promote apoptosis compared to those that facilitate protein folding and adaptation. As a consequence, the expression of apoptotic proteins is short-lived as cells adapt to stress. We provide evidence that the selective persistence of ER chaperone expression is also applicable to at least one instance of genetic ER stress. This work provides new insight into how a stress response pathway can be structured to allow cells to avert death as they adapt. It underscores the contribution of posttranscriptional and posttranslational mechanisms in influencing this outcome

Transcriptional Profiling of Chondrodysplasia Growth Plate Cartilage Reveals Adaptive ER-Stress Networks That Allow Survival but Disrupt Hypertrophy

Metaphyseal chondrodysplasia, Schmid type (MCDS) is characterized by mild short stature and growth plate hypertrophic zone expansion, and caused by collagen X mutations. We recently demonstrated the central importance of ER stress in the pathology of MCDS by recapitulating the disease phenotype by expressing misfolding forms of collagen X (Schmid) or thyroglobulin (Cog) in the hypertrophic zone. Here we characterize the Schmid and Cog ER stress signaling networks by transcriptional profiling of microdissected mutant and wildtype hypertrophic zones. Both models displayed similar unfolded protein responses (UPRs), involving activation of canonical ER stress sensors and upregulation of their downstream targets, including molecular chaperones, foldases, and ER-associated degradation machinery. Also upregulated were the emerging UPR regulators Wfs1 and Syvn1, recently identified UPR components including Armet and Creld2, and genes not previously implicated in ER stress such as Steap1 and Fgf21. Despite upregulation of the Chop/Cebpb pathway, apoptosis was not increased in mutant hypertrophic zones. Ultrastructural analysis of mutant growth plates revealed ER stress and disrupted chondrocyte maturation throughout mutant hypertrophic zones. This disruption was defined by profiling the expression of wildtype growth plate zone gene signatures in the mutant hypertrophic zones. Hypertrophic zone gene upregulation and proliferative zone gene downregulation were both inhibited in Schmid hypertrophic zones, resulting in the persistence of a proliferative chondrocyte-like expression profile in ER-stressed Schmid chondrocytes. Our findings provide a transcriptional map of two chondrocyte UPR gene networks in vivo, and define the consequences of UPR activation for the adaptation, differentiation, and survival of chondrocytes experiencing ER stress during hypertrophy. Thus they provide important insights into ER stress signaling and its impact on cartilage pathophysiology

C-Terminal Region of EBNA-2 Determines the Superior Transforming Ability of Type 1 Epstein-Barr Virus by Enhanced Gene Regulation of LMP-1 and CXCR7

Type 1 Epstein-Barr virus (EBV) strains immortalize B lymphocytes in vitro much more efficiently than type 2 EBV, a difference previously mapped to the EBNA-2 locus. Here we demonstrate that the greater transforming activity of type 1 EBV correlates with a stronger and more rapid induction of the viral oncogene LMP-1 and the cell gene CXCR7 (which are both required for proliferation of EBV-LCLs) during infection of primary B cells with recombinant viruses. Surprisingly, although the major sequence differences between type 1 and type 2 EBNA-2 lie in N-terminal parts of the protein, the superior ability of type 1 EBNA-2 to induce proliferation of EBV-infected lymphoblasts is mostly determined by the C-terminus of EBNA-2. Substitution of the C-terminus of type 1 EBNA-2 into the type 2 protein is sufficient to confer a type 1 growth phenotype and type 1 expression levels of LMP-1 and CXCR7 in an EREB2.5 cell growth assay. Within this region, the RG, CR7 and TAD domains are the minimum type 1 sequences required. Sequencing the C-terminus of EBNA-2 from additional EBV isolates showed high sequence identity within type 1 isolates or within type 2 isolates, indicating that the functional differences mapped are typical of EBV type sequences. The results indicate that the C-terminus of EBNA-2 accounts for the greater ability of type 1 EBV to promote B cell proliferation, through mechanisms that include higher induction of genes (LMP-1 and CXCR7) required for proliferation and survival of EBV-LCLs

The construction and characterization of bovine herpesvirus 1 expressing cytokines

This thesis investigated the potential of using Bovine Herpesvirus-1 (BHV-1) virus as a viral vector to express cytokines, and analysed the effects of these cytokines on BHV-1 infection and host immunity. Two constructs were generated by homologous recombination by using transfer vectors containing either EL-1β or bovine IFN-γ flanked by the 5' and 3' ends of glycoprotein C gene. Recombination occurred within the gC locus which created a recombinant virus with a gC minus (gC -) background phenotype that expressed the cytokine using the gC promoter. Molecular characterization showed that the recombinant cytokine genes were expressed with similar kinetics to a late BHV-1 protein; mRNA expression was detected at 5 hour post-infection followed by the detection of biologically active protein. These recombinant proteins had biological activity comparable to a recombinant protein standard. Both recombinant viruses, BHV1/1L1β and BHV-1/IFNγ, exhibited 'in vitro' growth characteristics similar to a gC-minus virus. This indicated that the expression, of these cytokine did not affect BHV-1 growth. To analysis the 'in vivo' effects of these recombinant viruses, two experimental models were used. First, cattle were used to analyse the immune responses and viral pathogenicity during infection by recombinant BHV-1/IFNγ. Similar levels of virus shedding and similar clinical responses were observed for both recombinant BHV-1/IFNγ and gC-/LacZ+ viruses. BHV-1 was shown to be a potent inducer of boIFN-γ in the nasal cavity and IFN-γ secretion correlated with the onset and duration of viral shedding. Analysis of cellular and humoral responses did not reveal any significant immune modulation during infection by BHV-1/IFN-γ. Serum IgG, mucosal IgA and antibody viruneutralization titers were similar between BHV-1/IFNγ and control gC-/LacZ+ virus. In addition, gD specific proliferative responses and IFN-γ ELISPOTs did not reveal any differences. After re-activation from latency with dexamethasone, viral shedding was similar for both BHV-1/IFNγ and gC-/LacZ+ viruses. These results suggested that during a respiratory infection, the production of exogenous IFN-γ did not provide an advantage to the host. Also, BHV-1/IFN-γ was a stable vector during latency and recrudescence and expressed biologically active IFN-γ protein throughout the experimental period. A sheep model for BHV-1 infection was also used to evaluate the recombinant BHV-1 vector. A preliminary experiment showed that both wild-type BHV-1 Cooper virus and gC-/LacZ+ infected sheep, caused mild clinical signs and generated BHV-1 specific immune responses. Recombinant BHV-1/IL-1β and BHV-1/IFNγ were then used to infect sheep. In this experiment, there was no difference in viral pathogenesis or the immune responses to the recombinant viruses and gC-/LacZ+ virus. Results in both sheep and cattle indicated that using BHV-1 as a viral vector to express bovine IFN-γ or IL-1β did not alter pathogenesis or the host immune response