Metabolic flux technology with the Seahorse bioanalyzer has emerged as a standard technique in cellular metabolism studies, allowing for simultaneous kinetic measurements of respiration and glycolysis. Methods to extend the utility and versatility of the metabolic flux assay would undoubtedly have immediate and wide-reaching impacts. Herein, we describe a platform that couples the metabolic flux assay with high-content fluorescence imaging to simultaneously provide means for normalization of respiration data with cell number; analyze cell cycle distribution; and quantify mitochondrial content, fragmentation state, membrane potential, and mitochondrial reactive oxygen species. Integration of fluorescent dyes directly into the metabolic flux assay generates a more complete data set of mitochondrial features in a single assay. Moreover, application of this integrated strategy revealed insights into mitochondrial function following PGC1a and PRC1 inhibition in pancreatic cancer and demonstrated how the Rho-GTPases impact mitochondrial dynamics in breast cancer.
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http://dx.doi.org/10.1038/s42003-020-0988-z | DOI Listing |
J Biomater Sci Polym Ed
December 2024
Department of Pharmaceutics, Dr. Prabhakar B Kore Basic Science Research Center, Off-campus, KLE College of Pharmacy (A constituent unit of KAHER-Belagavi), Bengaluru, Karnataka, India.
Zaltoprofen (ZAL) is a non-steroidal anti-inflammatory drug (NSAID) with a short half-life (∼2.8 h) due to extensive first pass metabolism. In this context, 16 different polymeric film forming solutions (PFFS) of ZAL were developed using different grades of Eudragits, Polyvinylpyrrolidones, Kollicoat MAE 100 P and Hydroxypropyl cellulose as film formers, and polyethylene glycol 400 as a plasticizer in equal parts of ethanol and isopropyl alcohol used as solvents.
View Article and Find Full Text PDFJ Hazard Mater
December 2024
School of Civil Engineering and Architecture, Taizhou University, Taizhou 318000, China. Electronic address:
The removal of selenite (Se(IV)) and cadmium (Cd(II)) from low-carbon wastewater presents significant challenges. However, the addition of external organic carbon sources is limited in application due to the high cost and potential for secondary pollution. This study introduced a "hibernation-like microbial survival strategy", enabling efficient removal of Se(IV) and Cd(II) in sulfur autotrophic reactor, with S acting as the electron donor.
View Article and Find Full Text PDFCurr Opin Biotechnol
December 2024
Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical and Biomolecular Engineering (BK21 four), KAIST Institute for BioCentury, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 34141, Republic of Korea; Systems Metabolic Engineering and Systems Healthcare Cross-Generation Collaborative Laboratory, KAIST, Daejeon 34141, Republic of Korea; BioProcess Engineering Research Center, KAIST, Daejeon 34141, Republic of Korea; Graduate School of Engineering Biology, KAIST, Daejeon 34141, Republic of Korea; Center for Synthetic Biology, KAIST, Daejeon 34141, Republic of Korea. Electronic address:
The escalating climate crisis underscores the urgent need for sustainable development, with CO utilization emerging as a pivotal approach to mitigating greenhouse gas emissions. Among various technological approaches, metabolic engineering of microorganisms for CO utilization offers significant potential. This review covers the engineering of endogenous CO fixation pathways, the construction of novel synthetic pathways, and strategies to optimize metabolic flux, enhance cofactor availability, and manipulate regulatory genes to improve CO assimilation efficiency.
View Article and Find Full Text PDFActa Neuropathol Commun
December 2024
Laboratory of Neurological Infections and Immunity, National Institute of Allergy and Infectious Diseases, Division of Intramural Research, Rocky Mountain Laboratories, National Institutes of Health, Hamilton, MT, USA.
Misfolding of normal prion protein (PrP) to pathological isoforms (prions) causes prion diseases (PrDs) with clinical manifestations including cognitive decline and mood-related behavioral changes. Cognition and mood are linked to the neurophysiology of the limbic system. Little is known about how the disease affects the synaptic activity in brain parts associated with this system.
View Article and Find Full Text PDFNeurobiol Dis
December 2024
Department of Bioengineering, University of Maryland, College Park, MD 20742, United States of America. Electronic address:
Niemann Pick Disease Type C (NP-C), a rare neurogenetic disease with no known cure, is caused by mutations in the cholesterol trafficking protein NPC1. Brain microvascular endothelial cells (BMEC) are thought to play a critical role in the pathogenesis of several neurodegenerative diseases; however, little is known about how these cells are altered in NP-C. In this study, we investigated how NPC1 inhibition perturbs BMEC metabolism in human induced pluripotent stem cell-derived BMEC (hiBMEC).
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