Image based annotation of Chemogenomic Libraries for Phenotypic Screening

Phenotypical screening is a widely used approach in drug discovery for the identification of small molecules with cellular activities. However, functional annotation of identified hits often poses a challenge. The development of small molecules with narrow or exclusive target selectivity such as chemical probes and chemogenomic (CG) libraries, greatly diminishes this challenge, but non-specific effects caused by compound toxicity or interference with basic cellular functions still poses a problem to associate phenotypic readouts with molecular targets. Hence, each compound should ideally be comprehensively characterized regarding its effects on general cell functions. Here, we report an optimized live-cell multiplexed assay that classifies cells based on nuclear morphology, presenting an excellent indicator for cellular responses such as early apoptosis and necrosis. This basic readout in combination with the detection of other general cell damaging activities of small molecules such as changes in cytoskeletal morphology, cell cycle and mitochondrial health provides a comprehensive time-dependent characterization of the effect of small molecules on cellular health in a single experiment. The developed high-content assay offers multi-dimensional comprehensive characterization that can be used to delineate generic effects regarding cell functions and cell viability, allowing an assessment of compound suitability for subsequent detailed phenotypic and mechanistic studies.

Comprehensive Characterization And Clinical Relevance of CLEC4G Expression Across Human Cancers

Liver and lymph node sinusoidal endothelial cell C-type lectin (CLEC4G) interacts with the surface glycoproteins of enveloped viruses and mediates immune evasion for viruses. Recent studies have indicated that the expression of CLEC4G affects the development and microenvironment of tumors. In the present study, we revealed comprehensive characterization of CLEC4G expression across human cancers. We first explored the effect of CLEC4G expression on overall survival (OS) across human cancers. High expression of CLEC4G was beneficial in lung adenocarcinoma (LUAD) and detrimental in kidney renal clear cell carcinoma (KIRC) and uveal melanoma (UVM) in terms of OS. Enrichment analysis of the 14 cancer-related signatures suggested that CLEC4G regulated tumor metastasis and immune response. Enrichment analysis of the immunophenotypes and cancer-immunity cycles indicated that CLEC4G was involved in regulation of macrophages and Treg cells, which affected tumor immune response. We also observed correlations between CLEC4G expression and checkpoint molecular expression in different cancer types. Taken together, the present study revealed the comprehensive characterization of CLEC4G expression across human cancers, predicting the roles of CLEC4G in the development and immune microenvironment of tumors.

Responsiveness to Students’ Mathematical Thinking in Middle-Grades Classrooms

Responsiveness to students’ mathematical thinking is a characteristic of classroom discourse that reflects the extent to which students’ mathematical ideas are present, attended to, and taken up as the basis for instruction. Using the Mathematically Responsive Interaction (MRI) Framework and data from 11 middle-grades classrooms, we illustrate varied enactments of responsiveness and describe fluctuations in and relationships among different components of responsiveness. We found positive associations between different components of responsiveness, but they were not entirely predictive of one another. Individual classrooms appeared more or less responsive depending on which component was foregrounded. Our findings offer a more comprehensive characterization of responsiveness that documents the intertwined nature of teacher moves and student contributions during all whole-class instruction.