flowClust: a Bioconductor package for automated gating of flow cytometry data

Background  

As a high-throughput technology that offers rapid quantification of multidimensional characteristics for millions of cells, flow cytometry (FCM) is widely used in health research, medical diagnosis and treatment, and vaccine development. Nevertheless, there is an increasing concern about the lack of appropriate software tools to provide an automated analysis platform to parallelize the high-throughput data-generation platform. Currently, to a large extent, FCM data analysis relies on the manual selection of sequential regions in 2-D graphical projections to extract the cell populations of interest. This is a time-consuming task that ignores the high-dimensionality of FCM data.     

Methods for predicting bacterial protein subcellular localization

The computational prediction of the subcellular localization of bacterial proteins is an important step in genome annotation and in the search for novel vaccine or drug targets. Since the 1991 release of PSORT I--the first comprehensive algorithm to predict bacterial protein localization--many other localization prediction tools have been developed. These methods offer significant improvements in predictive performance over PSORT I and the accuracy of some methods now rivals that of certain high-throughput laboratory methods for protein localization identification.     

FuGEFlow: data model and markup language for flow cytometry

Background  

Flow cytometry technology is widely used in both health care and research. The rapid expansion of flow cytometry applications has outpaced the development of data storage and analysis tools. Collaborative efforts being taken to eliminate this gap include building common vocabularies and ontologies, designing generic data models, and defining data exchange formats. The Minimum Information about a Flow Cytometry Experiment (MIFlowCyt) standard was recently adopted by the International Society for Advancement of Cytometry. This standard guides researchers on the information that should be included in peer reviewed publications, but it is insufficient for data exchange and integration between computational systems. The Functional Genomics Experiment (FuGE) formalizes common aspects of comprehensive and high throughput experiments across different biological technologies. We have extended FuGE object model to accommodate flow cytometry data and metadata.     

Rpp25 is a major target of autoantibodies to the Th/To complex as measured by a novel chemiluminescent assay

Introduction

Autoantibodies to the Th/To antigen have been described in systemic sclerosis (SSc) and several proteins of the macromolecular Th/To complex have been reported to react with anti-Th/To antibodies. However, anti-Th/To has not been clinically utilized due to unavailability of commercial tests. The objective of the present study is to evaluate the newly developed ELISA and chemiluminescent immunoassay (CLIA) to measure autoantibodies to Rpp25 (a component of the Th/To complex) using immunoprecipitation (IP) as the reference method.

Methods

The first cohort consisted of 123 SSc patients including 7 anti-Th/To positive samples confirmed by IP. Additional seven anti-Th/To positive samples from non-SSc patients were also tested. For evaluation of the QUANTA Flash Rpp25 CLIA (research use only), 8 anti-Th/To IP positives, a cohort of 70 unselected SSc patients and sera from various disease controls (n = 357) and random healthy individuals (n = 10) were studied.

Results

Anti-Rpp25 antibodies determined by ELISA were found in 11/14 anti-Th/To IP positive but only in 1/156 (0.6%) negative samples resulting in a positive percent agreement of 78.6% (95% confidence interval [CI] 49.2, 95.3%) and a negative percent agreement of 99.4% (95% CI 96.4, 100.0%). To verify the results using a second method, 53 samples were tested by ELISA and CLIA for anti-Rpp25 reactivity and the results were highly correlated (rho = 0.71, 95% CI 0.56, 0.81; P < 0.0001). To define the cutoff of the CLIA, anti-Th/To IP positive and negative sera were tested using the anti-Rpp25 CLIA. At the cutoff selected by receiver operating characteristic (ROC) analysis 8/8 (100.0%) of the anti-Th/To positive sera but only 2/367 (0.5%) of the controls were positive for anti-Rpp25 antibodies. The positive and negative percent agreements were 100.0% (95% CI 63.1, 100.0%) and 99.5% (95% CI 98.0, 99.9%), respectively. In the disease cohorts 2/70 (2.9%) of the SSc patients were positive for anti-Rpp25 antibodies compared to 2/367 (0.5%) of the controls (P = 0.032). ROC analysis showed discrimination between SSc patients and controls with an area under the curve value of 0.732 (95% CI 0.655, 0.809).

Conclusion

Rpp25 is a major target of autoantibodies to the Th/To autoantigen complex. Further studies are needed to evaluate the clinical utility of the new assays.     

Behavioral response of polar bears to aircraft activity on the northern coast of Alaska

The rapid loss of arctic sea ice is forcing a larger proportion of the Southern Beaufort Sea polar bear (Ursus maritimus) population to spend more time on land, increasing chances of negative interactions between people and bears. In the United States, the Marine Mammal Protection Act (MMPA) protects polar bears from incidental disturbance from human activities. For the remote and roadless areas of northern Alaska, USA, effective management of small aircraft activity is necessary to limit disturbance, but effects of overflights on polar bear behavior are largely unknown. During 2021 and 2022, we intentionally exposed polar bears (n = 115) to systematic aircraft activity (helicopter, fixed-wing) until we observed a disruption of behavior that qualified as a level B take response (e.g., abrupt change in activity or movement) under the MMPA. We used a Bayesian logistic regression to determine what factors influence and can be used to predict when a polar bear will exhibit a level B take response and estimate the probability of an aircraft eliciting a level B take response at different altitudes above the polar bear. Aircraft type, flight altitude, landscape (barrier islands vs. mainland), and bear behavior (active vs. inactive) upon initial aircraft encounter were all important predictors of take. Probability of take rapidly increased with a decrease in flight altitude starting at 450 m for helicopter and 300 m for fixed-wing aircraft. Active (e.g., standing, walking) polar bears on barrier-island landscapes were more likely to experience take than inactive (e.g., bedded) bears on mainland landscapes. Our findings can help with assessments and management plans by quantifying disturbance to polar bears from current and future human activity that involves aircraft use.     

Workshop on the ecosystem and fisheries of the Coral Sea: an Australian perspective on research and management

This report summarizes a workshop on the Coral Sea to determine key research findings and identify the research gaps needed to support sustainable management of a proposed Coral Sea Marine Reserve. Key research questions included determining the connectivity of apex predators with the broader southwest Pacific Ocean, and assessing the regions?? biodiversity in relation to seabed topography and oceanographic processes. The workshop concluded noting the importance of engaging surrounding countries in maintaining the sustainability and uniqueness of the Coral Sea.     

Glycomacropeptide, a low-phenylalanine protein isolated from cheese whey, supports growth and attenuates metabolic stress in the murine model of phenylketonuria

Phenylketonuria (PKU) is caused by a mutation in the phenylalanine (phe) hydroxylase gene and requires a low-phe diet plus amino acid (AA) formula to prevent cognitive impairment. Glycomacropeptide (GMP) contains minimal phe and provides a palatable alternative to AA formula. Our objective was to compare growth, body composition, and energy balance in Pah(enu2) (PKU) and wild-type mice fed low-phe GMP, low-phe AA, or high-phe casein diets from 3-23 wk of age. The 2 × 2 × 3 design included main effects of genotype, sex, and diet. Fat and lean mass were assessed by dual-energy X-ray absorptiometry, and acute energy balance was assessed by indirect calorimetry. PKU mice showed growth and lean mass similar to wild-type littermates fed the GMP or AA diets; however, they exhibited a 3-15% increase in energy expenditure, as reflected in oxygen consumption, and a 3-30% increase in food intake. The GMP diet significantly reduced energy expenditure, food intake, and plasma phe concentration in PKU mice compared with the casein diet. The high-phe casein diet or the low-phe AA diet induced metabolic stress in PKU mice, as reflected in increased energy expenditure and intake of food and water, increased renal and spleen mass, and elevated plasma cytokine concentrations consistent with systemic inflammation. The low-phe GMP diet significantly attenuated these adverse effects. Moreover, total fat mass, %body fat, and the respiratory exchange ratio (CO(2) produced/O(2) consumed) were significantly lower in PKU mice fed GMP compared with AA diets. In summary, GMP provides a physiological source of low-phe dietary protein that promotes growth and attenuates the metabolic stress induced by a high-phe casein or low-phe AA diet in PKU mice.     

The Effect of Perspective on Presence and Space Perception

In this paper we report two experiments in which the effect of perspective projection on presence and space perception was investigated. In Experiment 1, participants were asked to score a presence questionnaire when looking at a virtual classroom. We manipulated the vantage point, the viewing mode (binocular versus monocular viewing), the display device/screen size (projector versus TV) and the center of projection. At the end of each session of Experiment 1, participants were asked to set their preferred center of projection such that the image seemed most natural to them. In Experiment 2, participants were asked to draw a floor plan of the virtual classroom. The results show that field of view, viewing mode, the center of projection and display all significantly affect presence and the perceived layout of the virtual environment. We found a significant linear relationship between presence and perceived layout of the virtual classroom, and between the preferred center of projection and perceived layout. The results indicate that the way in which virtual worlds are presented is critical for the level of experienced presence. The results also suggest that people ignore veridicality and they experience a higher level of presence while viewing elongated virtual environments compared to viewing the original intended shape.