A novel method for high-throughput detection and quantification of neutrophil extracellular traps reveals ROS-independent NET release with immune complexes

AbstractA newly-described first-line immune defence mechanism of neutrophils is the release of neutrophil extracellular traps (NETs). Immune complexes (ICxs) induce low level NET release. As such, the in vitro quantification of NETs is challenging with current methodologies. In order to investigate the role of NET release in ICx-mediated autoimmune diseases, we developed a highly sensitive and automated method for quantification of NETs. After labelling human neutrophils with PKH26 and extracellular DNA with Sytox green, cells are fixed and automatically imaged with 3-dimensional confocal laser scanning microscopy (3D-CLSM). NET release is then quantified with digital image analysis whereby the NET amount (Sytox green area) is corrected for the number of imaged neutrophils (PKH26 area). A high sensitivity of the assay is achieved by a) significantly augmenting the area of the well imaged (11%) as compared to conventional assays (0.5%) and b) using a 3D imaging technique for optimal capture of NETs, which are topologically superimposed on neutrophils. In this assay, we confirmed low levels of NET release upon human ICx stimulation which were positive for citrullinated histones and neutrophil elastase. In contrast to PMA-induced NET release, ICx-induced NET release was unchanged when co-incubated with diphenyleneiodonium (DPI). We were able to quantify NET release upon stimulation with serum from RA and SLE patients, which was not observed with normal human serum. To our knowledge, this is the first semi-automated assay capable of sensitive detection and quantification of NET release at a low threshold by using 3D CLSM. The assay is applicable in a high-throughput manner and allows the in vitro analysis of NET release in ICx-mediated autoimmune diseases

Symbiosis between the TRECVid benchmark and video libraries at the Netherlands Institute for Sound and Vision

Audiovisual archives are investing in large-scale digitisation efforts of their analogue holdings and, in parallel, ingesting an ever-increasing amount of born- digital files in their digital storage facilities. Digitisation opens up new access paradigms and boosted re-use of audiovisual content. Query-log analyses show the shortcomings of manual annotation, therefore archives are complementing these annotations by developing novel search engines that automatically extract information from both audio and the visual tracks. Over the past few years, the TRECVid benchmark has developed a novel relationship with the Netherlands Institute of Sound and Vision (NISV) which goes beyond the NISV just providing data and use cases to TRECVid. Prototype and demonstrator systems developed as part of TRECVid are set to become a key driver in improving the quality of search engines at the NISV and will ultimately help other audiovisual archives to offer more efficient and more fine-grained access to their collections. This paper reports the experiences of NISV in leveraging the activities of the TRECVid benchmark

High-level feature detection from video in TRECVid: a 5-year retrospective of achievements

Successful and effective content-based access to digital video requires fast, accurate and scalable methods to determine the video content automatically. A variety of contemporary approaches to this rely on text taken from speech within the video, or on matching one video frame against others using low-level characteristics like colour, texture, or shapes, or on determining and matching objects appearing within the video. Possibly the most important technique, however, is one which determines the presence or absence of a high-level or semantic feature, within a video clip or shot. By utilizing dozens, hundreds or even thousands of such semantic features we can support many kinds of content-based video navigation. Critically however, this depends on being able to determine whether each feature is or is not present in a video clip. The last 5 years have seen much progress in the development of techniques to determine the presence of semantic features within video. This progress can be tracked in the annual TRECVid benchmarking activity where dozens of research groups measure the effectiveness of their techniques on common data and using an open, metrics-based approach. In this chapter we summarise the work done on the TRECVid high-level feature task, showing the progress made year-on-year. This provides a fairly comprehensive statement on where the state-of-the-art is regarding this important task, not just for one research group or for one approach, but across the spectrum. We then use this past and on-going work as a basis for highlighting the trends that are emerging in this area, and the questions which remain to be addressed before we can achieve large-scale, fast and reliable high-level feature detection on video

Cognitive emotion regulation strategies and depressive symptoms: differences between males and females.

FSW - Self-regulation models for health behavior and Psychopathology - Ou

Cognitive coping strategies and symptoms of depression and anxiety: A comparison between adolescents and adults.

FSW - Self-regulation models for health behavior and Psychopathology - Ou

Neurodevelopmental evaluation and referral practices in children with congenital heart disease in central South Africa

Introduction: Children with congenital heart disease (CHD) are at higher risk for developmental delays than the general population. The American Heart Association (AHA) published a guideline to address these concerns in 2012. This study determined the neurodevelopmental evaluation and referral practices of practitioners in central South Africa.Method: An online survey was administered to practitioners (n=45) including paediatric cardiologists (n=4), cardiothoracic surgeons (n=4) and general paediatricians (n=37). Information on practitioner characteristics, awareness of the 2012 AHA guideline; and neurodevelopmental evaluation and referral practices was collected.Results: Twenty-one practitioners responded, including paediatric cardiologists (n=4), cardiothoracic surgeons (n=2) and paediatricians (n=15). Data for 20 practitioners was included. Despite most practitioners (n=18) indicating guidelines for the management of development were important, the majority (n=16; 80%) were unaware of the guideline. Most practitioners (n=18; 90%) failed to risk stratify children to identify those to be evaluated. Children with developmental delays were referred for formal developmental evaluation (n=11; 55%) and to intervention therapies (n= 15; 75%).Conclusion: Most practitioners are unaware of the 2012 AHA guideline. Awareness of the developmental risks associated with CHD and implementation of the guideline could promote early identification of developmental delays with referral to intervention therapies

The relationship between cognitive emotion regulation strategies and emotional problems: comparison between a clinical and a non-clinical sample.

FSW - Self-regulation models for health behavior and Psychopathology - Ou

Establishing the role of rare coding variants in known Parkinson's disease risk loci

Many common genetic factors have been identified to contribute to Parkinson's disease (PD) susceptibility, improving our understanding of the related underlying biological mechanisms. The involvement of rarer variants in these loci has been poorly studied. Using International Parkinson's Disease Genomics Consortium data sets, we performed a comprehensive study to determine the impact of rare variants in 23 previously published genome-wide association studies (GWAS) loci in PD. We applied Prix fixe to select the putative causal genes underneath the GWAS peaks, which was based on underlying functional similarities. The Sequence Kernel Association Test was used to analyze the joint effect of rare, common, or both types of variants on PD susceptibility. All genes were tested simultaneously as a gene set and each gene individually. We observed a moderate association of common variants, confirming the involvement of the known PD risk loci within our genetic data sets. Focusing on rare variants, we identified additional association signals for LRRK2, STBD1, and SPATA19. Our study suggests an involvement of rare variants within several putatively causal genes underneath previously identified PD GWAS peaks