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Full-Text Articles in Medicine and Health Sciences

Il-1r And Myd88 Contribute To The Absence Of A Bacterial Microbiome On The Healthy Murine Cornea, Stephanie J. Wan, Aaron B. Sullivan, Peyton Shieh, Matteo M. E. Metruccio, David J. Evans, Carolyn R. Bertozzi, Suzanne M. J. Fleiszig Jan 2018

Il-1r And Myd88 Contribute To The Absence Of A Bacterial Microbiome On The Healthy Murine Cornea, Stephanie J. Wan, Aaron B. Sullivan, Peyton Shieh, Matteo M. E. Metruccio, David J. Evans, Carolyn R. Bertozzi, Suzanne M. J. Fleiszig

Faculty Publications & Research of the TUC College of Pharmacy

Microbial communities are important for the health of mucosal tissues. Traditional culture and gene sequencing have demonstrated bacterial populations on the conjunctiva. However, it remains unclear if the cornea, a transparent tissue critical for vision, also hosts a microbiome. Corneas of wild-type, IL-1R (-/-) and MyD88 (-/-) C57BL/6 mice were imaged after labeling with alkyne-functionalized D-alanine (alkDala), a probe that only incorporates into the peptidoglycan of metabolically active bacteria. Fluorescence in situ hybridization (FISH) was also used to detect viable bacteria. AlkDala labeling was rarely observed on healthy corneas. In contrast, adjacent conjunctivae harbored filamentous alkDala-positive forms, that also labeled …


Piwi Is Required To Limit Exhaustion Of Aging Somatic Stem Cells, Pedro Sousa-Victor, Arshad Ayyaz, Rippei Hayashi, Yanyan Qi, David T. Madden, Victoria V. Lunyak, Heinrich Jasper Jan 2017

Piwi Is Required To Limit Exhaustion Of Aging Somatic Stem Cells, Pedro Sousa-Victor, Arshad Ayyaz, Rippei Hayashi, Yanyan Qi, David T. Madden, Victoria V. Lunyak, Heinrich Jasper

Faculty Publications & Research of the TUC College of Pharmacy

Please see the graphical abstract in the supplemental files.


Contributions Of Myd88-Dependent Receptors And Cd11c-Positive Cells To Corneal Epithelial Barrier Function Against Pseudomonas Aeruginosa, Matteo M. E. Metruccio, Connie Tam, David J. Evans, Anna L. Xie, Michael E. Stern, Suzanne M. J. Fleiszig Jan 2017

Contributions Of Myd88-Dependent Receptors And Cd11c-Positive Cells To Corneal Epithelial Barrier Function Against Pseudomonas Aeruginosa, Matteo M. E. Metruccio, Connie Tam, David J. Evans, Anna L. Xie, Michael E. Stern, Suzanne M. J. Fleiszig

Faculty Publications & Research of the TUC College of Pharmacy

Previously we reported that corneal epithelial barrier function against Pseudomonas aeruginosa was MyD88-dependent. Here, we explored contributions of MyD88-dependent receptors using vital mouse eyes and confocal imaging. Uninjured IL-1R (−/−) or TLR4 (−/−) corneas, but not TLR2 (−/−), TLR5 (−/−), TLR7 (−/−), or TLR9 (−/−), were more susceptible to P. aeruginosa adhesion than wild-type (3.8-fold, 3.6-fold respectively). Bacteria adherent to the corneas of IL-1R (−/−) or TLR5 (−/−) mice penetrated beyond the epithelial surface only if the cornea was superficially-injured. Bone marrow chimeras showed that bone marrow-derived cells contributed to IL-1R-dependent barrier function. In vivo, but not ex vivo …


Determining Maximum Glycolytic Capacity Using Extracellular Flux Measurements, Shona A. Mookerjee, David G. Nicholls, Martin D. Brand Jan 2016

Determining Maximum Glycolytic Capacity Using Extracellular Flux Measurements, Shona A. Mookerjee, David G. Nicholls, Martin D. Brand

Faculty Publications & Research of the TUC College of Pharmacy

Measurements of glycolytic rate and maximum glycolytic capacity using extracellular flux analysis can give crucial information about cell status and phenotype during normal operation, development of pathology, differentiation, and malignant transformation. They are also of great use when assessing the effects of chemical or drug treatments. Here, we experimentally define maximum glycolytic capacity, demonstrate how it differs from glycolytic rate, and provide a protocol for determining the basal glycolytic rate and maximum glycolytic capacity in cells using extracellular flux measurements. The results illustrate the power of extracellular flux analysis to describe the energetics of adherent cells in culture in a …


Measurement Of The Absolute Magnitude And Time Courses Of Mitochondrial Membrane Potential In Primary And Clonal Pancreatic Beta-Cells, Akos Gerencser, Shona A. Mookerjee, Martin Jastroch, Martin D. Brand Jan 2016

Measurement Of The Absolute Magnitude And Time Courses Of Mitochondrial Membrane Potential In Primary And Clonal Pancreatic Beta-Cells, Akos Gerencser, Shona A. Mookerjee, Martin Jastroch, Martin D. Brand

Faculty Publications & Research of the TUC College of Pharmacy

The aim of this study was to simplify, improve and validate quantitative measurement of the mitochondrial membrane potential (ΔψM) in pancreatic β-cells. This built on our previously introduced calculation of the absolute magnitude of ΔψM in intact cells, using time-lapse imaging of the non-quench mode fluorescence of tetramethylrhodamine methyl ester and a bis-oxonol plasma membrane potential (ΔψP) indicator. ΔψM is a central mediator of glucose-stimulated insulin secretion in pancreatic β-cells. ΔψM is at the crossroads of cellular energy production and demand, therefore precise assay of its magnitude is a valuable tool to study how these processes interplay in insulin secretion. …