Notes: Hypoxic cancer cells constitute a major challenge in chemo- and radiotherapy. The researchers present evaluation of APD-CLD compounds that induce selective cell death in hypoxic cancer cells by impeding mitochondrial function. The RealTime-Glo™ Annexin V Apoptosis Assay, CellTiter-Blue® Cell Viability Assay, CellTox™ Green Real-Time Cytotoxicity Assay and GSH-Glo™ Glutathione Assay were used to assess mechanism of action. (5168)

Notes: These authors describe the design, assembly and toxicant response of multi-cellular 3D hepatic organotypic culture models. In high-throughput screening, changes in cell viability were determined over 24 hours using the RealTime-Glo™ MT Cell Viability Assay. Mechanism of cell death was determined with the ApoTox-Glo™ Triplex Assay, and GSH concentration was measured using the GSH-Glo™ Glutathione Assay.  (5011)

Notes: The authors of this study explore the hypothesis that mitochondria pass glycolytic enzymes to the nucleus during early development to assist with producing substrates needed for epigenetic changes needed for activating the zygotic genome. Embryo extracts were analyzed with the ENLITEN® ATP Assay and the ADP-Glo™ Assay for ATP:ADP ratio; glutathione levels monitored with GSH-Glo™ Assay.  Extracts were also made as directed in the NAD/NADH-Glo™ Assay manual to determine NAD and NADH levels. (4831)

Notes: The authors tested if CYP1B1 removed cellular oxygenation products that induce oxidative stress and promote the release of antiangiogenic factors. The P450-Glo™ CYP1B1 Assay was used to determine CYP1B1 activity. The presence of glutathione was assessed using either 10⁴ retinal endothelial cells or 50µl of mouse retinal extracts dispensed into each well of a 96-well plate with the GSH-Glo™ Glutathione Assay. (4010)

Notes: Glutathione levels were assessed in human coronary artery endothelial cells (HCAECs) as a measure of oxidative stress. HCAECs were treated with either 100ng/ml eotaxin, a newly discovered chemokine, or pretreated with 2µmol/l MnTBAP for 30 minutes followed by eotaxin treatment for 45 minutes. Positive controls were treatment with 10µg/ml antimycin A and 2ng/ml TNF-α. Cellular glutathione was measured using the GSH-Glo™ Glutathione Assay. (4011)

Notes: In this study, the researchers wanted to determine the role of kelch-like ECH associated protein 1 knockdown (Keap1-kd) mice protein products, which are thought to protect against oxidative and electrophilic stress, and compare the hepatic phenotype with that of transcription factor nuclear factor erythroid 2–related factor 2 (Nrf2)-null and wild-type mice. Microsomal suspensions from liver homogenates were prepared, and bile was collected from wild-type, Nrf2-null, and Keap1-kd mice. Reduced GSH was quantified using the GSH-Glo™ Glutathione Assay. (4012)

Notes: The authors of this review article highlight the use of bioluminescence as a readout for high-throughput ADME/Tox assays. They discuss three strategies for designing bioluminescent assays, using either luciferase, ATP or luciferin substrates as the limiting reagents for a luciferase-catalyzed reaction. Reporter gene assays limit the production of luciferase by tying it to a promoter or DNA regulatory region of interest. Such assays can be used to study genes that are regulated by drugs and other xenobiotics. Bioluminescent assays in which ATP is the limiting reagent of the luciferase reaction can be designed to monitor cell viability or the activity kinases. Bioluminescent assays in which the substrate is limiting can be designed so that the activity of a particular enzyme results in the production of a luciferin substrate that can, in turn, be acted upon by luciferase. (3926)