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1.  Islet β-Cell Endoplasmic Reticulum Stress Precedes the Onset of Type 1 Diabetes in the Nonobese Diabetic Mouse Model 
Diabetes  2012;61(4):818-827.
Type 1 diabetes is preceded by islet β-cell dysfunction, but the mechanisms leading to β-cell dysfunction have not been rigorously studied. Because immune cell infiltration occurs prior to overt diabetes, we hypothesized that activation of inflammatory cascades and appearance of endoplasmic reticulum (ER) stress in β-cells contributes to insulin secretory defects. Prediabetic nonobese diabetic (NOD) mice and control diabetes-resistant NOD-SCID and CD1 strains were studied for metabolic control and islet function and gene regulation. Prediabetic NOD mice were relatively glucose intolerant and had defective insulin secretion with elevated proinsulin:insulin ratios compared with control strains. Isolated islets from NOD mice displayed age-dependent increases in parameters of ER stress, morphologic alterations in ER structure by electron microscopy, and activation of nuclear factor-κB (NF-κB) target genes. Upon exposure to a mixture of proinflammatory cytokines that mimics the microenvironment of type 1 diabetes, MIN6 β-cells displayed evidence for polyribosomal runoff, a finding consistent with the translational initiation blockade characteristic of ER stress. We conclude that β-cells of prediabetic NOD mice display dysfunction and overt ER stress that may be driven by NF-κB signaling, and strategies that attenuate pathways leading to ER stress may preserve β-cell function in type 1 diabetes.
doi:10.2337/db11-1293
PMCID: PMC3314371  PMID: 22442300
2.  Optical Nanosensor Architecture for Cell Signaling Molecules Using DNA Aptamer-Coated Carbon Nanotubes 
ACS nano  2011;5(5):4236-4244.
We report a novel optical biosensor platform using near-infrared (NIR) fluorescent single-walled carbon nanotubes (SWNTs) functionalized with target-recognizing aptamer DNA for noninvasively detecting cell signaling molecules in real-time. Photoluminescence (PL) emission of aptamer-coated SWNTs is modulated upon selectively binding to target molecules, which is exploited to detect insulin using an insulin-binding aptamer (IBA) as a molecular recognition element. We find that nanotube PL quenches upon insulin recognition via a photoinduced charge transfer mechanism with a quenching rate of kq = 5.85×1014 M−1s−1 and a diffusion-reaction rate of kr = 0.129 s−1. Circular dichroism spectra reveal for the first time that IBA strands retain a four-stranded, parallel guanine quadruplex conformation on the nanotubes, ensuring target selectivity. We demonstrate that these IBA-functionalized SWNT sensors incorporated in a collagen extracellular matrix (ECM) can be regenerated by removing bound analytes through enzymatic proteolysis. As proof-of-concept, we show that the SWNT sensors embedded in the ECM promptly detect insulin secreted by cultured pancreatic INS-1 cells stimulated by glucose influx and report a gradient contour of insulin secretion profile. This novel design enables new types of label-free assays and non-invasive, in-situ, real-time detection schemes for cell signaling molecules.
doi:10.1021/nn201323h
PMCID: PMC3178844  PMID: 21520951
Aptamers; Biosensors; Carbon Nanotubes; Insulin; Near-infrared Fluorescence
3.  Oscillatory glucose flux in INS 1 pancreatic β cells: A self-referencing microbiosensor study 
Analytical biochemistry  2010;411(2):185-193.
Signaling and insulin secretion in β cells have been reported to demonstrate oscillatory modes, with abnormal oscillations associated with type 2 diabetes. We investigated cellular glucose influx in β cells with a self-referencing microbiosensor based on nanomaterials with enhanced performance. Dose-response analysis with glucose and metabolic inhibition studies were used to study oscillatory pattern and transporter kinetics. For the first time, we report a stable and regular oscillatory uptake of glucose (averaged period 2.9±0.6minutes), which corresponds well with an oscillator model. This oscillatory behavior is part of the feedback control pathway involving oxygen, cytosolic Ca2+/ATP, and insulin secretion (periodicity approximately 3 minutes). Glucose stimulation experiments show that the net Michaelis-Menten constant (6.1±1.5mM) is in between GLUT2 and GLUT9. Phloretin inhibition experiments show an EC50 value of 28±1.6μM-phloretin for class I GLUT proteins and a concentration of 40±0.6μM-phloretin caused maximum inhibition with residual non-oscillating flux, suggesting that the transporters not inhibited by phloretin are likely responsible for the remaining non-oscillatory uptake, and that impaired uptake via GLUT2 may be the cause of the oscillation loss in type 2 diabetes. Transporter studies using the SR microbiosensor will contribute to diabetes research and therapy development by exploring the nature of oscillatory transport mechanisms.
doi:10.1016/j.ab.2010.12.019
PMCID: PMC3081878  PMID: 21167120
glucose oxidase; biosensor; self referencing; diabetes; glycolysis; β cell
4.  The role of multiscale computational approaches for rational design of conventional and nanoparticle oral drug delivery systems 
Multiscale computational modeling of drug delivery systems (DDS) is poised to provide predictive capabilities for the rational design of targeted drug delivery systems, including multi-functional nanoparticles. Realistic, mechanistic models can provide a framework for understanding the fundamental physico-chemical interactions between drug, delivery system, and patient. Multiscale computational modeling, however, is in its infancy even for conventional drug delivery. The wide range of emerging nanotechnology systems for targeted delivery further increases the need for reliable in silico predictions. This review will present existing computational approaches at different scales in the design of traditional oral drug delivery systems. Subsequently, a multiscale framework for integrating continuum, stochastic, and computational chemistry models will be proposed and a case study will be presented for conventional DDS. The extension of this framework to emerging nanotechnology delivery systems will be discussed along with future directions. While oral delivery is the focus of the review, the outlined computational approaches can be applied to other drug delivery systems as well.
PMCID: PMC2676650  PMID: 18019831
Oral drug delivery; multiscale; computational modeling; continuum; computational chemistry; stochastic

Results 1-4 (4)