ABSTRACT
BACKGROUND: Neurite outgrowth is a metric widely used to assess the success of in vitro neural stem cell differentiation or neuron reprogramming protocols and to evaluate high-content screening assays for neural regenerative drug discovery. However, neurite measurements are tedious to perform manually, and there is a paucity of freely available, fully automated software to determine neurite measurements and neuron counting. To provide such a tool to the neurobiology, stem cell, cell engineering, and neuroregenerative communities, we developed an algorithm for performing high-throughput neurite analysis in immunofluorescent images. NEW METHOD: Given an input of paired neuronal nuclear and cytoskeletal microscopy images, the GAIN algorithm calculates neurite length statistics linked to individual cells or clusters of cells. It also provides an estimate of the number of nuclei in clusters of overlapping cells, thereby increasing the accuracy of neurite length statistics for higher confluency cultures. GAIN combines image processing for neuronal cell bodies and neurites with an algorithm for resolving neurite junctions. RESULTS: GAIN produces a table of neurite lengths from cell body to neurite tip per cell cluster in an image along with a count of cells per cluster. COMPARISON WITH EXISTING METHODS: GAIN's performance compares favorably with the popular ImageJ plugin NeuriteTracer for counting neurons, and provides the added benefit of assigning neurites to their respective cell bodies. CONCLUSIONS: In summary, GAIN provides a new tool to improve the robust assessment of neural cells by image-based analysis.
Subject(s)
Cell Tracking/methods , Neural Stem Cells/cytology , Neural Stem Cells/physiology , Neurites/physiology , Neurites/ultrastructure , Neuronal Outgrowth/physiology , Pattern Recognition, Automated/methods , Algorithms , Animals , Cells, Cultured , Image Interpretation, Computer-Assisted/methods , Mice , Reproducibility of Results , Sensitivity and Specificity , Subtraction TechniqueABSTRACT
Forty-four women and three men who were plaintiffs in sexual harassment and sexual discrimination cases were evaluated by experts for the plaintiffs and by the author, an expert in these cases, for the defense. There were considerable differences in the diagnostic assessments. Some reasons for these differences and their implications for forensic evaluations are discussed.
Subject(s)
Expert Testimony/legislation & jurisprudence , Mental Disorders/diagnosis , Sexual Harassment/legislation & jurisprudence , Female , Humans , Male , Mental Disorders/classification , Mental Disorders/psychology , Observer Variation , Personality Assessment/statistics & numerical data , PsychometricsABSTRACT
Twenty-three girls and thirteen boys, aged between 4 and 9 years, gave urine samples which were measured by HPLC for adrenaline, noradrenaline, cortisol and cortisone on a school day and on a home day. Home levels of noradrenaline, cortisol and cortisone were consistently higher than school levels. Cortisol was positively correlated with adrenaline and noradrenaline at home, but not at school. These results may suggest that the school day represents a stimulating but comparatively structured environment with little negative affect, while the home day is characterised by stimulation but relative uncertainty in social interactions.
