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.

In a pre-post-treatment design, this feasibility study examines the effects of twelve weekly online Feldenkrais Awareness Through Movement (ATM) lessons on somatosensory tinnitus (ST), which is often associated with physical issues like neck pain. Two professional Feldenkrais teachers led online ATM lessons in two groups for a total of seventeen participants. Before and after treatment, we assessed tinnitus distress (using the Tinnitus Questionnaire, TQ), interoceptive awareness, and the short WHO quality-of-life questionnaire. We collected weekly changes in tinnitus severity (using the Tinnitus Functional Index, TFI) and neck pain (using the Neck Bournemouth Questionnaire), and assessed state anxiety and perceived tinnitus intensity directly before and after each ATM. Importantly, we found a significant reduction in tinnitus distress along with significant individual differences. Linear mixed-model analyses further suggest an overall decrease in tinnitus severity and neck pain over time. Notably, individual reductions in TFI correlated with reduced neck pain, and reductions in TQ correlated with increased interoceptive awareness. Although this study is limited in patient number, it provides valuable information about the characteristics of patients with ST and suggests a new method to reduce neck pain and perceived tinnitus in the comfort of patients’ homes. However, further studies are necessary to verify these results and to compare the effectiveness with conventional therapy approaches.