D6.2: Intermediate Standardisation and Dissemination Activity Report

This report documents the standardisation and dissemination activities performed by the ORPHEUS project consortium from December 2015 to February 2017

bogJS – A JavaScript framework for object-based rendering in browsers

Presented at the 2nd Web Audio Conference (WAC), April 4-6, 2016, Atlanta, Georgia.With the introduction of HTML5 and the Web Audio API, an important prerequisite was made for native rendering of object-based audio in modern browsers. Object-based audio is a revolutionary approach for creating and deploying interactive, personalized, scalable and immersive content, by representing it as a set of individual assets together with metadata describing their relationships and associations. This allows media objects to be assembled in ground-breaking ways to create new user experiences. This talk will introduce the open-source framework bogJS, developed by IRT (Institut für Rundfunktechnik) within the scope of the EU funded project ORPHEUS, which utilizes native nodes of the Web Audio API to realize such experiences. One aim of the development was to provide a flexible API that can be easily extended with different types of user interfaces and various representations of object metadata. Another key aim of the framework is to offer different possibilities to access audio signals for the rendering by taking into account current drawbacks and limitations of browser such as the supported number of tracks in a media element, the ordering of decoded multi-channel tracks or synchronized playback issues. Interoperability and implementation of current and upcoming standards, such as the ITU-R BS.2076 recommendation for a metadata definition model, will be taken into account and are currently under development. Furthermore, first projects and demonstrations using the framework will be presented

bogJS v0.4.0 (object-based renderer for web browsers)

<p>JavaScript framework for object-based audio rendering in modern browsers from IRT. Parts of this framework were developed in the European collaborative research project ORPHEUS.</p

Comparison of Microbiomes from Different Niches of Upper and Lower Airways in Children and Adolescents with Cystic Fibrosis

Changes in the airway microbiome may be important in the pathophysiology of chronic lung disease in patients with cystic fibrosis. However, little is known about the microbiome in early cystic fibrosis lung disease and the relationship between the microbiomes from different niches in the upper and lower airways. Therefore, in this cross-sectional study, we examined the relationship between the microbiome in the upper (nose and throat) and lower (sputum) airways from children with cystic fibrosis using next generation sequencing. Our results demonstrate a significant difference in both α and β-diversity between the nose and the two other sampling sites. The nasal microbiome was characterized by a polymicrobial community while the throat and sputum communities were less diverse and dominated by a few operational taxonomic units. Moreover, sputum and throat microbiomes were closely related especially in patients with clinically stable lung disease. There was a high inter-individual variability in sputum samples primarily due to a decrease in evenness linked to increased abundance of potential respiratory pathogens such as Pseudomonas aeruginosa. Patients with chronic Pseudomonas aeruginosa infection exhibited a less diverse sputum microbiome. A high concordance was found between pediatric and adult sputum microbiomes except that Burkholderia was only observed in the adult cohort. These results indicate that an adult-like lower airways microbiome is established early in life and that throat swabs may be a good surrogate in clinically stable children with cystic fibrosis without chronic Pseudomonas aeruginosa infection in whom sputum sampling is often not feasible

NOX1 Supports the Metabolic Remodeling of HepG2 Cells

<div><p>NADPH oxidases are important sources of reactive oxygen species (ROS) which act as signaling molecules in the regulation of protein expression, cell proliferation, differentiation, migration and cell death. The NOX1 subunit is over-expressed in several cancers and NOX1 derived ROS have been repeatedly linked with tumorigenesis and tumor progression although underlying pathways are ill defined. We engineered NOX1-depleted HepG2 hepatoblastoma cells and employed differential display 2DE experiments in order to investigate changes in NOX1-dependent protein expression profiles. A total of 17 protein functions were identified to be dysregulated in NOX1-depleted cells. The proteomic results support a connection between NOX1 and the Warburg effect and a role for NOX in the regulation of glucose and glutamine metabolism as well as of lipid, protein and nucleotide synthesis in hepatic tumor cells. Metabolic remodeling is a common feature of tumor cells and understanding the underlying mechanisms is essential for the development of new cancer treatments. Our results reveal a manifold involvement of NOX1 in the metabolic remodeling of hepatoblastoma cells towards a sustained production of building blocks required to maintain a high proliferative rate, thus rendering NOX1 a potential target for cancer therapy.</p></div

2DE map displaying proteins differentially expressed in control vs. NOX1 reduced HepG2 cells.

<p>Empty arrows denote up-regulated protein functions, filled arrows denote down-regulated protein functions. White arrows denote differentially regulated but un-identified protein spots.</p

Western blot validation of NOX1 regulated proteins.

<p>The major components of the 2DE spots which contained complex proteins mixtures were subjected to validation through Western blot analysis. Seven of the proteins were confirmed to be differentially expressed in control vs. NOX1 reduced HepG2 cells. Plotted are the mean and SEM of WB intensities normalized to the total protein content of the sample determined from the PonceauS staining. SET—protein SET, HMGC—HMG-CoA-synthase, AFP—Alpha-fetoprotein, Acsl1—Long chain fatty acid-CoA ligase, GDH1—Glutamate dehydrogenase 1, UDPGP—UTP-glucose-1-phosphate uridylyltransferase. Statistical results are presented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0122002#pone.0122002.t001" target="_blank">Table 1</a>.</p

NOX1 regulates glycogen content in HepG2 cells.

<p>Glycogen levels were assessed in HepG2 transiently transfected with two shRNAs against NOX1 or with control shRNA. NOX1 reduced HepG2 cells produce more glycogen than shCtr expressing cells (right). Differences in glycogen levels were tested in a linear mixed effect model with cell line as predictor and replicate (N = 4) as random factor (one-tailed post-hoc z test). NOX1 depletion was confirmed be Western blot analysis (left).</p

Overview of the metabolic pathways regulated by NOX1.

<p>Proteins differentially expressed in control vs. NOX1 reduced HepG2 cells are involved in glucose, glutamine, nucleotide and lipid metabolism. α-KG—α-ketoglutarate, OAA—oxaloacetic acid, PEP—phosphoenolpyruvate, TAG—triacylglycerol, TCA cycle—tricarboxylic acid cycle.</p

Abundance of differentially expressed proteins depends upon NOX1 relative expression (plotted are 95% CI).

<p>Abundance of differentially expressed proteins depends upon NOX1 relative expression (plotted are 95% CI).</p