Improved standardization of flow cytometry diagnostic screening of primary immunodeficiency by software-based automated gating

BackgroundMultiparameter flow cytometry (FC) is essential in the diagnostic work-up and classification of primary immunodeficiency (PIDs). The EuroFlow PID Orientation tube (PIDOT) allows identification of all main lymphocyte subpopulations in blood. To standardize data analysis, tools for Automated Gating and Identification (AG&I) of the informative cell populations, were developed by EuroFlow. Here, we evaluated the contribution of these innovative AG&I tools to the standardization of FC in the diagnostic work-up of PID, by comparing AG&I against expert-based (EuroFlow-standardized) Manual Gating (MG) strategy, and its impact on the reproducibility and clinical interpretation of results.MethodsFC data files from 44 patients (13 CVID, 12 PID, 19 non-PID) and 26 healthy donor (HD) blood samples stained with PIDOT were analyzed in parallel by MG and AG&I, using Infinicyt (TM) software (Cytognos). For comparison, percentage differences in absolute cell counts/mu L were calculated for each lymphocyte subpopulation. Data files showing differences >20% were checked for their potential clinical relevance, based on age-matched percentile (p5-p95) reference ranges. In parallel, intra- and inter-observer reproducibility of MG vs AG&I were evaluated in a subset of 12 samples.ResultsThe AG&I approach was able to identify the vast majority of lymphoid events (>99%), associated with a significantly higher intra- and inter-observer reproducibility compared to MG. For most HD (83%) and patient (68%) samples, a high degree of agreement (<20% numerical differences in absolute cell counts/mu L) was obtained between MG and the AG&I module. This translated into a minimal impact (<5% of observations) on the final clinical interpretation. In all except three samples, extended expert revision of the AG&I approach revealed no error. In the three remaining samples aberrant maturation and/or abnormal marker expression profiles were seen leading in all three cases to numerical alarms by AG&I.ConclusionAltogether, our results indicate that replacement of MG by the AG&I module would be associated with a greater reproducibility and robustness of results in the diagnostic work-up of patients suspected of PID. However, expert revision of the results of AG&I of PIDOT data still remains necessary in samples with numerical alterations and aberrant B- and T-cell maturation and/or marker expression profiles.Stemcel biology/Regenerative medicine (incl. bloodtransfusion

Project Phantom

Report for the course IN3700 (Bachelor project). In this case the project was to make a game called Project Phantom. The project was in collaboration with CannibalGameStudios which supplied the project and the tutoring.MKTElectrical Engineering, Mathematics and Computer Scienc

Impaired phagocytic activity of neutrophils in patients receiving haemodialysis: the critical role of iron overload.

The metabolic burst (as measured by the spontaneous and stimulated nitroblue tetrazolium tests), the phagocytosis of heat inactivated bakers' yeast and of Staphylococcus aureus, the killing of Staph aureus, and the myeloperoxidase activity of polymorphonuclear neutrophils were studied in 11 patients receiving maintenance haemodialysis. Of these patients, six were polytransfused and had high serum ferritin concentrations (mean 5940 (SD 2925) micrograms/l; group 1), and five had normal serum ferritin values (mean 171 (116) micrograms/l; group 2). Patients in group 1 had a history of more infectious episodes (0.167 v 0.025 per patient per month) and significantly more genitourinary infections (p = 0.015) than those in group 2. Phagocytosis and myeloperoxidase activity were severely reduced in group 1 but normal in group 2. Percentages of neutrophils ingesting one or more particles together with the index of phagocytosis in patients' serum were inversely correlated with serum ferritin concentrations. Four patients in group 1 were treated with desferrioxamine, and after six to 18 weeks of treatment phagocytosis and myeloperoxidase activity had returned to normal in three of them. These data suggest that in patients receiving haemodialysis iron overload due to multiple transfusions plays an important part in the mechanisms underlying the susceptibility to bacterial infections, mediated at least partially through impaired neutrophil function

Corporate Twitter Channels: The Impact of Engagement and Informedness on Corporate Reputation

We examine communication via a corporate Twitter channel and its effects on corporate reputation. We identify the importance of user engagement and informedness in explaining corporate reputation and examine three design factors that likely affect user engagement in a corporate Twitter channel. We conduct an exploratory 2 x 2 x 2 experiment among Twitter users to collect data. We find that the depth of the relationship among users, the level of corporate involvement, and the purpose of the channel interactively influence user engagement. Our findings suggest that deeper relationships among users of a corporate Twitter channel lead to higher user engagement when the level of corporate involvement with the channel is high and when the channel has a specific purpose, but not when the level of corporate involvement is high and the channel has a generic purpose. Surprisingly, when the channel has a generic purpose, a high degree of corporate involvement actually decreases user engagement. This finding implies that, under certain circumstances, a lower degree of corporate involvement in a social media channel may be more desirable. We also find that channel credibility positively influences user informedness. This is the first study that examines the dynamics of communication through a corporate Twitter channel. It contributes to the previous research related to social media by identifying engagement and informedness as two major factors that influence firms' reputation. Our research can help marketing and social media managers to decide on channel design aspects, such as whether to require users to register with an identity or to allow anonymous participation, whether to allocate dedicated employees to respond to user requests, and whether to set up different channels for different purposes