O-GlcNAcylation of Orphan Nuclear Receptor Estrogen-Related Receptor γ Promotes Hepatic Gluconeogenesis.

Science.gov (United States)

Misra, Jagannath; Kim, Don-Kyu; Jung, Yoon Seok; Kim, Han Byeol; Kim, Yong-Hoon; Yoo, Eun-Kyung; Kim, Byung Gyu; Kim, Sunghoon; Lee, In-Kyu; Harris, Robert A; Kim, Jeong-Sun; Lee, Chul-Ho; Cho, Jin Won; Choi, Hueng-Sik

2016-10-01

Estrogen-related receptor γ (ERRγ) is a major positive regulator of hepatic gluconeogenesis. Its transcriptional activity is suppressed by phosphorylation signaled by insulin in the fed state, but whether posttranslational modification alters its gluconeogenic activity in the fasted state is not known. Metabolically active hepatocytes direct a small amount of glucose into the hexosamine biosynthetic pathway, leading to protein O-GlcNAcylation. In this study, we demonstrate that ERRγ is O-GlcNAcylated by O-GlcNAc transferase in the fasted state. This stabilizes the protein by inhibiting proteasome-mediated protein degradation, increasing ERRγ recruitment to gluconeogenic gene promoters. Mass spectrometry identifies two serine residues (S317, S319) present in the ERRγ ligand-binding domain that are O-GlcNAcylated. Mutation of these residues destabilizes ERRγ protein and blocks the ability of ERRγ to induce gluconeogenesis in vivo. The impact of this pathway on gluconeogenesis in vivo was confirmed by the observation that decreasing the amount of O-GlcNAcylated ERRγ by overexpressing the deglycosylating enzyme O-GlcNAcase decreases ERRγ-dependent glucose production in fasted mice. We conclude that O-GlcNAcylation of ERRγ serves as a major signal to promote hepatic gluconeogenesis. © 2016 by the American Diabetes Association.

Electron-transfer mediator for a NAD-glucose dehydrogenase-based glucose sensor.

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Kim, Dong-Min; Kim, Min-yeong; Reddy, Sanapalli S; Cho, Jaegeol; Cho, Chul-ho; Jung, Suntae; Shim, Yoon-Bo

2013-12-03

A new electron-transfer mediator, 5-[2,5-di (thiophen-2-yl)-1H-pyrrol-1-yl]-1,10-phenanthroline iron(III) chloride (FePhenTPy) oriented to the nicotinamide adenine dinucleotide-dependent-glucose dehydrogenase (NAD-GDH) system was synthesized through a Paal-Knorr condensation reaction. The structure of the mediator was confirmed by Fourier-transform infrared spectroscopy, proton and carbon nucler magnetic resonance spectroscopy, and mass spectroscopy, and its electron-transfer characteristic for a glucose sensor was investigated using voltammetry and impedance spectroscopy. A disposable amperometric glucose sensor with NAD-GDH was constructed with FePhenTPy as an electron-transfer mediator on a screen printed carbon electrode (SPCE) and its performance was evaluated, where the addition of reduces graphene oxide (RGO) to the mediator showed the enhanced sensor performance. The experimental parameters to affect the analytical performance and the stability of the proposed glucose sensor were optimized, and the sensor exhibited a dynamic range between 30 mg/dL and 600 mg/dL with the detection limit of 12.02 ± 0.6 mg/dL. In the real sample experiments, the interference effects by acetaminophen, ascorbic acid, dopamine, uric acid, caffeine, and other monosaccharides (fructose, lactose, mannose, and xylose) were completely avoided through coating the sensor surface with the Nafion film containing lead(IV) acetate. The reliability of proposed glucose sensor was evaluated by the determination of glucose in artificial blood and human whole blood samples.

Glucose phosphorylation is not rate limiting for accumulation of glycogen from glucose in perfused livers from fasted rats

International Nuclear Information System (INIS)

Youn, J.H.; Ader, M.; Bergman, R.N.

1989-01-01

Incorporation of Glc and Fru into glycogen was measured in perfused livers from 24-h fasted rats using [6-3H]Glc and [U-14C]Fru. For the initial 20 min, livers were perfused with low Glc (2 mM) to deplete hepatic glycogen and were perfused for the following 30 min with various combinations of Glc and Fru. With constant Fru (2 mM), increasing perfusate Glc increased the relative contribution of Glc carbons to glycogen (7.2 +/- 0.4, 34.9 +/- 2.8, and 59.1 +/- 2.7% at 2, 10, and 20 mM Glc, respectively; n = 5 for each). During perfusion with substrate levels seen during refeeding (10 mM Glc, 1.8 mumol/g/min gluconeogenic flux from 2 mM Fru), Fru provided 54.7 +/- 2.7% of the carbons for glycogen, while Glc provided only 34.9 +/- 2.8%, consistent with in vivo estimations. However, the estimated rate of Glc phosphorylation was at least 1.10 +/- 0.11 mumol/g/min, which exceeded by at least 4-fold the glycogen accumulation rate (0.28 +/- 0.04 mumol of glucose/g/min). The total rate of glucose 6-phosphate supply via Glc phosphorylation and gluconeogenesis (2.9 mumol/g/min) exceeded reported in vivo rates of glycogen accumulation during refeeding. Thus, in perfused livers of 24-h fasted rats there is an apparent redundancy in glucose 6-phosphate supply. These results suggest that the rate-limiting step for hepatic glycogen accumulation during refeeding is located between glucose 6-phosphate and glycogen, rather than at the step of Glc phosphorylation or in the gluconeogenic pathway

Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity.

Science.gov (United States)

Shi, Hao; Munk, Alexander; Nielsen, Thomas S; Daughtry, Morgan R; Larsson, Louise; Li, Shize; Høyer, Kasper F; Geisler, Hannah W; Sulek, Karolina; Kjøbsted, Rasmus; Fisher, Taylor; Andersen, Marianne M; Shen, Zhengxing; Hansen, Ulrik K; England, Eric M; Cheng, Zhiyong; Højlund, Kurt; Wojtaszewski, Jørgen F P; Yang, Xiaoyong; Hulver, Matthew W; Helm, Richard F; Treebak, Jonas T; Gerrard, David E

2018-05-01

Given that cellular O-GlcNAcylation levels are thought to be real-time measures of cellular nutrient status and dysregulated O-GlcNAc signaling is associated with insulin resistance, we evaluated the role of O-GlcNAc transferase (OGT), the enzyme that mediates O-GlcNAcylation, in skeletal muscle. We assessed O-GlcNAcylation levels in skeletal muscle from obese, type 2 diabetic people, and we characterized muscle-specific OGT knockout (mKO) mice in metabolic cages and measured energy expenditure and substrate utilization pattern using indirect calorimetry. Whole body insulin sensitivity was assessed using the hyperinsulinemic euglycemic clamp technique and tissue-specific glucose uptake was subsequently evaluated. Tissues were used for histology, qPCR, Western blot, co-immunoprecipitation, and chromatin immunoprecipitation analyses. We found elevated levels of O-GlcNAc-modified proteins in obese, type 2 diabetic people compared with well-matched obese and lean controls. Muscle-specific OGT knockout mice were lean, and whole body energy expenditure and insulin sensitivity were increased in these mice, consistent with enhanced glucose uptake and elevated glycolytic enzyme activities in skeletal muscle. Moreover, enhanced glucose uptake was also observed in white adipose tissue that was browner than that of WT mice. Interestingly, mKO mice had elevated mRNA levels of Il15 in skeletal muscle and increased circulating IL-15 levels. We found that OGT in muscle mediates transcriptional repression of Il15 by O-GlcNAcylating Enhancer of Zeste Homolog 2 (EZH2). Elevated muscle O-GlcNAc levels paralleled insulin resistance and type 2 diabetes in humans. Moreover, OGT-mediated signaling is necessary for proper skeletal muscle metabolism and whole-body energy homeostasis, and our data highlight O-GlcNAcylation as a potential target for ameliorating metabolic disorders. Copyright © 2018 The Authors. Published by Elsevier GmbH.. All rights reserved.

Functional O-GlcNAc modifications: Implications in molecular regulation and pathophysiology

Science.gov (United States)

Wells, Lance

2016-01-01

O-linked β-N-acetylglucosamine (O-GlcNAc) is a regulatory post-translational modification of intracellular proteins. The dynamic and inducible cycling of the modification is governed by O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA) in response to UDP-GlcNAc levels in the hexosamine biosynthetic pathway (HBP). Due to its reliance on glucose flux and substrate availability, a major focus in the field has been on how O-GlcNAc contributes to metabolic disease. For years this post-translational modification has been known to modify thousands of proteins implicated in various disorders, but direct functional connections have until recently remained elusive. New research is beginning to reveal the specific mechanisms through which O-GlcNAc influences cell dynamics and disease pathology including clear examples of O-GlcNAc modification at a specific site on a given protein altering its biological functions. The following review intends to focus primarily on studies in the last half decade linking O-GlcNAc modification of proteins with chromatin-directed gene regulation, developmental processes, and several metabolically related disorders including Alzheimer’s, heart disease and cancer. These studies illustrate the emerging importance of this post-translational modification in biological processes and multiple pathophysiologies. PMID:24524620

Glycolysis-induced discordance between glucose metabolic rates measured with radiolabeled fluorodeoxyglucose and glucose

International Nuclear Information System (INIS)

Ackermann, R.F.; Lear, J.L.

1989-01-01

We have developed an autoradiographic method for estimating the oxidative and glycolytic components of local CMRglc (LCMRglc), using sequentially administered [ 18 F]fluorodeoxyglucose (FDG) and [ 14 C]-6-glucose (GLC). FDG-6-phosphate accumulation is proportional to the rate of glucose phosphorylation, which occurs before the divergence of glycolytic (GMg) and oxidative (GMo) glucose metabolism and is therefore related to total cerebral glucose metabolism GMt: GMg + GMo = GMt. With oxidative metabolism, the 14 C label of GLC is temporarily retained in Krebs cycle-related substrate pools. We hypothesize that with glycolytic metabolism, however, a significant fraction of the 14 C label is lost from the brain via lactate production and efflux from the brain. Thus, cerebral GLC metabolite concentration may be more closely related to GMo than to GMt. If true, the glycolytic metabolic rate will be related to the difference between FDG- and GLC-derived LCMRglc. Thus far, we have studied normal awake rats, rats with limbic activation induced by kainic acid (KA), and rats visually stimulated with 16-Hz flashes. In KA-treated rats, significant discordance between FDG and GLC accumulation, which we attribute to glycolysis, occurred only in activated limbic structures. In visually stimulated rats, significant discordance occurred only in the optic tectum

Conditions Inducing Excessive O-GlcNAcylation Inhibit BMP2-Induced Osteogenic Differentiation of C2C12 Cells.

Science.gov (United States)

Gu, Hanna; Song, Mina; Boonanantanasarn, Kanitsak; Baek, Kyunghwa; Woo, Kyung Mi; Ryoo, Hyun-Mo; Baek, Jeong-Hwa

2018-01-09

Hyperglycemic conditions in diabetic patients can affect various cellular functions, including the modulation of osteogenic differentiation. However, the molecular mechanisms by which hyperglycemia affects osteogenic differentiation are yet to be clarified. This study aimed to investigate whether the aberrant increase in protein O -linked-β- N -acetylglucosamine glycosylation ( O -GlcNAcylation) contributes to the suppression of osteogenic differentiation due to hyperglycemia. To induce osteogenic differentiation, C2C12 cells were cultured in the presence of recombinant human bone morphogenetic protein 2 (BMP2). Excessive protein O -GlcNAcylation was induced by treating C2C12 cells with high glucose, glucosamine, or N -acetylglucosamine concentrations or by O -GlcNAc transferase (OGT) overexpression. The effect of O -GlcNAcylation on osteoblast differentiation was then confirmed by examining the expression levels of osteogenic marker gene mRNAs, activity of alkaline phosphatase, and transcriptional activity of Runx2, a critical transcription factor for osteoblast differentiation and bone formation. Cell treatment with high glucose, glucosamine or N -acetylglucosamine increased O -GlcNAcylation of Runx2 and the total levels of O -GlcNAcylated proteins, which led to a decrease in the transcriptional activity of Runx2, expression levels of osteogenic marker genes (Runx2, osterix, alkaline phosphatase, and type I collagen), and activity of alkaline phosphatase. These inhibitory effects were rescued by lowering protein O -GlcNAcylation levels by adding STO45849, an OGT inhibitor, or by overexpressing β- N -acetylglucosaminidase. Our findings suggest that excessive protein O -GlcNAcylation contributes to high glucose-suppressed osteogenic differentiation.

Application of chronic intravascular blood glucose sensor in dogs.

Science.gov (United States)

Armour, J C; Lucisano, J Y; McKean, B D; Gough, D A

1990-12-01

An intravenous glucose sensor was implanted in six dogs for 1-15 wk. The glucose sensor is a flexible cylinder, approximately 0.2 cm diam and 30 cm long, with a tip containing immobilized glucose oxidase and catalase coupled to a potentiostatic O2 sensor. The sensor and a similar O2 reference sensor were implanted in the superior vena cava near the entrance of the right atrium. The sensor response was conveyed externally either by a telemetry system implanted nearby, surgically accessed leads, or chronically maintained percutaneous leads. Summing over the six implants, there was a total implantation period of 333 days during which glucose sensors were functional on demand. The sensor response showed agreement with conventionally assayed blood samples after accounting for a response lag. Sensor response to glucose showed little change over the implant period. Biocompatibility, enzyme lifetime, O2 availability, O2 sensor stability, and biochemical interference were not limitations. Results demonstrated that this sensor can function effectively as an implant in dogs for a period of months and has the potential for long-term operation.

Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity

Directory of Open Access Journals (Sweden)

Hao Shi

2018-05-01

Full Text Available Objective: Given that cellular O-GlcNAcylation levels are thought to be real-time measures of cellular nutrient status and dysregulated O-GlcNAc signaling is associated with insulin resistance, we evaluated the role of O-GlcNAc transferase (OGT, the enzyme that mediates O-GlcNAcylation, in skeletal muscle. Methods: We assessed O-GlcNAcylation levels in skeletal muscle from obese, type 2 diabetic people, and we characterized muscle-specific OGT knockout (mKO mice in metabolic cages and measured energy expenditure and substrate utilization pattern using indirect calorimetry. Whole body insulin sensitivity was assessed using the hyperinsulinemic euglycemic clamp technique and tissue-specific glucose uptake was subsequently evaluated. Tissues were used for histology, qPCR, Western blot, co-immunoprecipitation, and chromatin immunoprecipitation analyses. Results: We found elevated levels of O-GlcNAc-modified proteins in obese, type 2 diabetic people compared with well-matched obese and lean controls. Muscle-specific OGT knockout mice were lean, and whole body energy expenditure and insulin sensitivity were increased in these mice, consistent with enhanced glucose uptake and elevated glycolytic enzyme activities in skeletal muscle. Moreover, enhanced glucose uptake was also observed in white adipose tissue that was browner than that of WT mice. Interestingly, mKO mice had elevated mRNA levels of Il15 in skeletal muscle and increased circulating IL-15 levels. We found that OGT in muscle mediates transcriptional repression of Il15 by O-GlcNAcylating Enhancer of Zeste Homolog 2 (EZH2. Conclusions: Elevated muscle O-GlcNAc levels paralleled insulin resistance and type 2 diabetes in humans. Moreover, OGT-mediated signaling is necessary for proper skeletal muscle metabolism and whole-body energy homeostasis, and our data highlight O-GlcNAcylation as a potential target for ameliorating metabolic disorders. Keywords: O-GlcNAc signaling, Type 2 diabetes, N

Nutrient-driven O-GlcNAc in proteostasis and neurodegeneration.

Science.gov (United States)

Akan, Ilhan; Olivier-Van Stichelen, Stephanie; Bond, Michelle R; Hanover, John A

2018-01-01

Proteostasis is essential in the mammalian brain where post-mitotic cells must function for decades to maintain synaptic contacts and memory. The brain is dependent on glucose and other metabolites for proper function and is spared from metabolic deficits even during starvation. In this review, we outline how the nutrient-sensitive nucleocytoplasmic post-translational modification O-linked N-acetylglucosamine (O-GlcNAc) regulates protein homeostasis. The O-GlcNAc modification is highly abundant in the mammalian brain and has been linked to proteopathies, including neurodegenerative diseases such as Alzheimer's, Parkinson's, and Huntington's. C. elegans, Drosophila, and mouse models harboring O-GlcNAc transferase- and O-GlcNAcase-knockout alleles have helped define the role O-GlcNAc plays in development as well as age-associated neurodegenerative disease. These enzymes add and remove the single monosaccharide from protein serine and threonine residues, respectively. Blocking O-GlcNAc cycling is detrimental to mammalian brain development and interferes with neurogenesis, neural migration, and proteostasis. Findings in C. elegans and Drosophila model systems indicate that the dynamic turnover of O-GlcNAc is critical for maintaining levels of key transcriptional regulators responsible for neurodevelopment cell fate decisions. In addition, pathways of autophagy and proteasomal degradation depend on a transcriptional network that is also reliant on O-GlcNAc cycling. Like the quality control system in the endoplasmic reticulum which uses a 'mannose timer' to monitor protein folding, we propose that cytoplasmic proteostasis relies on an 'O-GlcNAc timer' to help regulate the lifetime and fate of nuclear and cytoplasmic proteins. O-GlcNAc-dependent developmental alterations impact metabolism and growth of the developing mouse embryo and persist into adulthood. Brain-selective knockout mouse models will be an important tool for understanding the role of O-GlcNAc in the

Graphene quantum dots prepared from glucose as optical sensor for glucose

Energy Technology Data Exchange (ETDEWEB)

Shehab, Mona, E-mail: mona_shehab@alexu.edu.eg [Materials Science Department, Institute of Graduate Studies & Research, Alexandria University (Egypt); General Bureau of Beheira Governorate, Damanhour, Beheira 22111 (Egypt); Ebrahim, Shaker; Soliman, Moataz [Materials Science Department, Institute of Graduate Studies & Research, Alexandria University (Egypt)

2017-04-15

Quantum Dots (QDs) show promise materials for many technological applications. In this work we utilized a simple route to prepare graphene quantum dots (GQDs) using glucose carbonization. GQDs functionalized with phenylboronic acid receptors were employed as a sensing material for a nonenzymatic glucose sensor. Photoluminance spectra of GQDs were used as a property of optical sensor for glucose. GQDs considered as a good sensing probe because of its low toxicity, high photoluminance, water solubility and excelent photochemical properties. The prepared GQDs were characterized using UV-visible, Raman and photoluminance spectroscopies, X-ray diffraction and high resolution transmission electron microscopy (HRTEM). HRTEM micrographs confirmed the preparation of 7–10 nm GQDs and the emission peak of the GQDs appeared at 450 nm. The developed sensor has linear response to glucose over a concentration range of 4–40 mM with a correlation coefficient of 0.97 and a low detection limit of approximately 3.0 mM.

Novel fungal FAD glucose dehydrogenase derived from Aspergillus niger for glucose enzyme sensor strips.

Science.gov (United States)

Sode, Koji; Loew, Noya; Ohnishi, Yosuke; Tsuruta, Hayato; Mori, Kazushige; Kojima, Katsuhiro; Tsugawa, Wakako; LaBelle, Jeffrey T; Klonoff, David C

2017-01-15

In this study, a novel fungus FAD dependent glucose dehydrogenase, derived from Aspergillus niger (AnGDH), was characterized. This enzyme's potential for the use as the enzyme for blood glucose monitor enzyme sensor strips was evaluated, especially by investigating the effect of the presence of xylose during glucose measurements. The substrate specificity of AnGDH towards glucose was investigated, and only xylose was found as a competing substrate. The specific catalytic efficiency for xylose compared to glucose was 1.8%. The specific activity of AnGDH for xylose at 5mM concentration compared to glucose was 3.5%. No other sugars were used as substrate by this enzyme. The superior substrate specificity of AnGDH was also demonstrated in the performance of enzyme sensor strips. The impact of spiking xylose in a sample with physiological glucose concentrations on the sensor signals was investigated, and it was found that enzyme sensor strips using AnGDH were not affected at all by 5mM (75mg/dL) xylose. This is the first report of an enzyme sensor strip using a fungus derived FADGDH, which did not show any positive bias at a therapeutic level xylose concentration on the signal for a glucose sample. This clearly indicates the superiority of AnGDH over other conventionally used fungi derived FADGDHs in the application for SMBG sensor strips. The negligible activity of AnGDH towards xylose was also explained on the basis of a 3D structural model, which was compared to the 3D structures of A. flavus derived FADGDH and of two glucose oxidases. Copyright © 2016 Elsevier B.V. All rights reserved.

Human Subcutaneous Tissue Response to Glucose Sensors: Macrophages Accumulation Impact on Sensor Accuracy.

Science.gov (United States)

Rigla, Mercedes; Pons, Belén; Rebasa, Pere; Luna, Alexis; Pozo, Francisco Javier; Caixàs, Assumpta; Villaplana, Maria; Subías, David; Bella, Maria Rosa; Combalia, Neus

2018-04-01

Subcutaneous (s.c.) glucose sensors have become a key component in type 1 diabetes management. However, their usability is limited by the impact of foreign body response (FBR) on their duration, reliability, and accuracy. Our study gives the first description of human acute and subacute s.c. response to glucose sensors, showing the changes observed in the sensor surface, the inflammatory cells involved in the FBR and their relationship with sensor performance. Twelve obese patients (seven type 2 diabetes) underwent two abdominal biopsies comprising the surrounding area where they had worn two glucose sensors: the first one inserted 7 days before and the second one 24 h before biopsy procedure. Samples were processed and studied to describe tissue changes by two independent pathologists (blind regarding sensor duration). Macrophages quantification was studied by immunohistochemistry methods in the area surrounding the sensor (CD68, CD163). Sensor surface changes were studied by scanning electron microscopy. Seven-day continuous glucose monitoring records were considered inaccurate when mean absolute relative difference was higher than 10%. Pathologists were able to correctly classify all the biopsies regarding sensor duration. Acute response (24 h) was characterized by the presence of neutrophils while macrophages were the main cell involved in subacute inflammation. The number of macrophages around the insertion hole was higher for less accurate sensors compared with those performing more accurately (32.6 ± 14 vs. 10.6 ± 1 cells/0.01 mm 2 ; P sensor-tissue interface is related with decrease in accuracy of the glucose measure.

Liver X receptor regulates hepatic nuclear O-GlcNAc signaling and carbohydrate responsive element-binding protein activity

DEFF Research Database (Denmark)

Bindesbøll, Christian; Fan, Qiong; Nørgaard, Rikke C

2015-01-01

in response to feeding, which is believed to be mediated by insulin. We have previously shown that LXRs are targets for glucose-hexosamine-derived O-linked β-N-acetylglucosamine (O-GlcNAc) modification enhancing their ability to regulate SREBP-1c promoter activity in vitro. To elucidate insulin...... of glycolytic and lipogenic enzymes, including glucokinase (GK), SREBP-1c, ChREBPα, and the newly identified shorter isoform ChREBPβ. Furthermore, glucose-dependent increases in LXR/retinoid X receptor-regulated luciferase activity driven by the ChREBPα promoter was mediated, at least in part, by O-GlcNAc...... transferase (OGT) signaling in Huh7 cells. Moreover, we show that LXR and OGT interact and colocalize in the nucleus and that loss of LXRs profoundly reduced nuclear O-GlcNAc signaling and ChREBPα promoter binding activity in vivo. In summary, our study provides evidence that LXRs act as nutrient and glucose...

Novel glucose fiber sensor combining ThFBG with GOD

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Li, Mengmeng; Zhou, Ciming; Fan, Dian; Ou, Yiwen

2016-10-01

We propose a novel glucose fiber optic sensor combining a thinned cladding fiber Bragg grating (ThFBG) with glucose oxidase (GOD). By immobilizing GOD on the surface of a ThFBG, the fabricated sensor can obtain a high specificity to glucose. Because of the evanescent field, the sensor is very sensitive to the ambient refractive index change arising from the catalytic reaction between glucose and GOD. A four-level fiber model was simulated and verified the precision of the sensing principle. Two methods, glutaraldehyde crosslinking method (GCM) and 3-aminopropyl triethoxysilane covalent coupling method (ATCCM), were experimentally utilized to immobilize GOD. And sensor fabricated with the method ATCCM shows a measurement range of 0-0.82 mg/mL which is better than the sensor fabricated with the method GCM with measurement range of 0-0.67 mg/mL under the same condition. By using ATCCM to immobilize GOD with different concentrations, three sensors were fabricated and used for glucose measurement by monitoring the Bragg wavelength (λb) shifts, the results indicate a good linear relationship between wavelength shift and glucose concentration within a specific range, and the measurement range increases as GOD concentration increases. The highest sensitivity of sensor reaches up to 0.0549 nm/(mg.mL-1). The proposed sensor has distinct advantages in sensing structure, cost and specificity.

Crosslinked basement membrane-based coatings enhance glucose sensor function and continuous glucose monitoring in vivo.

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Klueh, Ulrike; Ludzinska, Izabela; Czajkowski, Caroline; Qiao, Yi; Kreutzer, Donald L

2018-01-01

Overcoming sensor-induced tissue reactions is an essential element of achieving successful continuous glucose monitoring (CGM) in the management of diabetes, particularly when used in closed loop technology. Recently, we demonstrated that basement membrane (BM)-based glucose sensor coatings significantly reduced tissue reactions at sites of device implantation. However, the biocompatible BM-based biohydrogel sensor coating rapidly degraded over a less than a 3-week period, which effectively eliminated the protective sensor coating. In an effort to increase the stability and effectiveness of the BM coating, we evaluated the impact of crosslinking BM utilizing glutaraldehyde as a crosslinking agent, designated as X-Cultrex. Sensor performance (nonrecalibrated) was evaluated for the impact of these X-Cultrex coatings in vitro and in vivo. Sensor performance was assessed over a 28-day time period in a murine CGM model and expressed as mean absolute relative difference (MARD) values. Tissue reactivity of Cultrex-coated, X-Cultrex-coated, and uncoated glucose sensors was evaluated over a 28-day time period in vivo using standard histological techniques. These studies demonstrated that X-Cultrex-based sensor coatings had no effect on glucose sensor function in vitro. In vivo, glucose sensor performance was significantly enhanced following X-Cultrex coating throughout the 28-day study. Histological evaluations of X-Cultrex-treated sensors demonstrated significantly less tissue reactivity when compared to uncoated sensors. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 7-16, 2018. © 2017 Wiley Periodicals, Inc.

Glucose oxidase probe as a surface-enhanced Raman scattering sensor for glucose.

Science.gov (United States)

Qi, Guohua; Wang, Yi; Zhang, Biying; Sun, Dan; Fu, Cuicui; Xu, Weiqing; Xu, Shuping

2016-10-01

Glucose oxidase (GOx) possessing a Raman-active chromophore (flavin adenine dinucleotide) is used as a signal reporter for constructing a highly specific "turn off" surface-enhanced Raman scattering (SERS) sensor for glucose. This sensing chip is made by the electrostatic assembly of GOx over silver nanoparticle (Ag NP)-functionalized SERS substrate through a positively charged polyelectrolyte linker under the pH of 6.86. To trace glucose in blood serum, owing to the reduced pH value caused by the production of gluconic acid in the GOx-catalyzed oxidation reaction, the bonding force between GOx and polyelectrolyte weakens, making GOx drop off from the sensing chip. As a result, the SERS intensity of GOx on the chip decreases along with the concentration of glucose. This glucose SERS sensor exhibits excellent selectivity based on the specific GOx/glucose catalysis reaction and high sensitivity to 1.0 μM. The linear sensing range is 2.0-14.0 mM, which also meets the requirement on the working range of the human blood glucose detection. Using GOx as a probe shows superiority over other organic probes because GOx almost has no toxicity to the biological system. This sensing mechanism can be applied for intracellular in vivo SERS monitoring of glucose in the future. Graphical abstract Glucose oxidase is used as a Raman signal reporter for constructing a highly specific glucose surface-enhanced Raman scattering (SERS) sensor.

Increased O-GlcNAcylation of Endothelial Nitric Oxide Synthase Compromises the Anti-contractile Properties of Perivascular Adipose Tissue in Metabolic Syndrome.

Science.gov (United States)

da Costa, Rafael M; da Silva, Josiane F; Alves, Juliano V; Dias, Thiago B; Rassi, Diane M; Garcia, Luis V; Lobato, Núbia de Souza; Tostes, Rita C

2018-01-01

Under physiological conditions, the perivascular adipose tissue (PVAT) negatively modulates vascular contractility. This property is lost in experimental and human obesity and in the metabolic syndrome, indicating that changes in PVAT function may contribute to vascular dysfunction associated with increased body weight and hyperglycemia. The O -linked β-N-acetylglucosamine ( O -GlcNAc) modification of proteins ( O -GlcNAcylation) is a unique posttranslational process that integrates glucose metabolism with intracellular protein activity. Increased flux of glucose through the hexosamine biosynthetic pathway and the consequent increase in tissue-specific O -GlcNAc modification of proteins have been linked to multiple facets of vascular dysfunction in diabetes and other pathological conditions. We hypothesized that chronic consumption of glucose, a condition that progresses to metabolic syndrome, leads to increased O -GlcNAc modification of proteins in the PVAT, decreasing its anti-contractile effects. Therefore, the current study was devised to determine whether a high-sugar diet increases O -GlcNAcylation in the PVAT and how increased O -GlcNAc interferes with PVAT vasorelaxant function. To assess molecular mechanisms by which O -GlcNAc contributes to PVAT dysfunction, thoracic aortas surrounded by PVAT were isolated from Wistar rats fed either a control or high sugar diet, for 10 and 12 weeks. Rats chronically fed a high sugar diet exhibited metabolic syndrome features, increased O -GlcNAcylated-proteins in the PVAT and loss of PVAT anti-contractile effect. PVAT from high sugar diet-fed rats for 12 weeks exhibited decreased NO formation, reduced expression of endothelial nitric oxide synthase (eNOS) and increased O -GlcNAcylation of eNOS. High sugar diet also decreased OGA activity and increased superoxide anion generation in the PVAT. Visceral adipose tissue samples from hyperglycemic patients showed increased levels of O -GlcNAc-modified proteins, increased ROS

Estimation of glucose kinetics in fetal-maternal studies: Potential errors, solutions, and limitations

International Nuclear Information System (INIS)

Menon, R.K.; Bloch, C.A.; Sperling, M.A.

1990-01-01

We investigated whether errors occur in the estimation of ovine maternal-fetal glucose (Glc) kinetics using the isotope dilution technique when the Glc pool is rapidly expanded by exogenous (protocol A) or endogenous (protocol C) Glc entry and sought possible solutions (protocol B). In protocol A (n = 8), after attaining steady-state Glc specific activity (SA) by [U-14C]glucose (period 1), infusion of Glc (period 2) predictably decreased Glc SA, whereas. [U-14C]glucose concentration unexpectedly rose from 7,208 +/- 367 (means +/- SE) in period 1 to 8,558 +/- 308 disintegrations/min (dpm) per ml in period 2 (P less than 0.01). Fetal endogenous Glc production (EGP) was negligible during period 1 (0.44 +/- 1.0), but yielded a physiologically impossible negative value of -2.1 +/- 0.72 mg.kg-1.min-1 during period 2. When the fall in Glc SA during Glc infusion was prevented by addition of [U-14C]glucose admixed with the exogenous Glc (protocol B; n = 7), EGP was no longer negative. In protocol C (n = 6), sequential infusions of four increasing doses of epinephrine serially decreased SA, whereas tracer Glc increased from 7,483 +/- 608 to 11,525 +/- 992 dpm/ml plasma (P less than 0.05), imposing an obligatory underestimation of EGP. Thus a tracer mixing problem leads to erroneous estimations of fetal Glc utilization and Glc production via the three-compartment model in sheep when the Glc pool is expanded exogenously or endogenously. These errors can be minimized by maintaining the Glc SA relatively constant

Genetic interaction network of the Saccharomyces cerevisiae type 1 phosphatase Glc7

Directory of Open Access Journals (Sweden)

Neszt Michael

2008-07-01

Full Text Available Abstract Background Protein kinases and phosphatases regulate protein phosphorylation, a critical means of modulating protein function, stability and localization. The identification of functional networks for protein phosphatases has been slow due to their redundant nature and the lack of large-scale analyses. We hypothesized that a genome-scale analysis of genetic interactions using the Synthetic Genetic Array could reveal protein phosphatase functional networks. We apply this approach to the conserved type 1 protein phosphatase Glc7, which regulates numerous cellular processes in budding yeast. Results We created a novel glc7 catalytic mutant (glc7-E101Q. Phenotypic analysis indicates that this novel allele exhibits slow growth and defects in glucose metabolism but normal cell cycle progression and chromosome segregation. This suggests that glc7-E101Q is a hypomorphic glc7 mutant. Synthetic Genetic Array analysis of glc7-E101Q revealed a broad network of 245 synthetic sick/lethal interactions reflecting that many processes are required when Glc7 function is compromised such as histone modification, chromosome segregation and cytokinesis, nutrient sensing and DNA damage. In addition, mitochondrial activity and inheritance and lipid metabolism were identified as new processes involved in buffering Glc7 function. An interaction network among 95 genes genetically interacting with GLC7 was constructed by integration of genetic and physical interaction data. The obtained network has a modular architecture, and the interconnection among the modules reflects the cooperation of the processes buffering Glc7 function. Conclusion We found 245 genes required for the normal growth of the glc7-E101Q mutant. Functional grouping of these genes and analysis of their physical and genetic interaction patterns bring new information on Glc7-regulated processes.

Accuracy of a Fourth-Generation Subcutaneous Continuous Glucose Sensor.

Science.gov (United States)

Christiansen, Mark P; Garg, Satish K; Brazg, Ronald; Bode, Bruce W; Bailey, Timothy S; Slover, Robert H; Sullivan, Ashley; Huang, Suiying; Shin, John; Lee, Scott W; Kaufman, Francine R

2017-08-01

This study evaluated the accuracy and performance of a fourth-generation subcutaneous glucose sensor (Guardian ™ Sensor 3) in the abdomen and arm. Eighty-eight subjects (14-75 years of age, mean ± standard deviation [SD] of 42.0 ± 19.1 years) with type 1 or type 2 diabetes participated in the study. Subjects wore two sensors in the abdomen that were paired with either a MiniMed ™ 640G insulin pump, or an iPhone ® or iPod ® touch ® running a glucose monitoring mobile application (Guardian Connect system) and a third sensor in the arm, which was connected to a glucose sensor recorder (GSR). Subjects were also asked to undergo in-clinic visits of 12-14 h on study days 1, 3, and 7 for frequent blood glucose sample testing using a Yellow Springs Instrument (YSI) reference. The overall mean absolute relative difference (MARD ± SD) between abdomen sensor glucose (SG) and YSI reference values was 9.6% ± 9.0% and 9.4% ± 9.8% for the MiniMed 640G insulin pump and Guardian Connect system, respectively; and 8.7% ± 8.0% between arm SG and YSI reference values. The percentage of SG values within 20% agreement of the YSI reference value (for YSI >80 mg/dL) was 90.7% with the MiniMed 640G insulin pump, 91.8% with the Guardian Connect system, and 93.1% for GSR-connected arm sensors. Mean functional sensor life, when calibrating 3-4 times/day, was 145.9 ± 39.3 h for sensors paired with the MiniMed 640G insulin pump, 146.1 ± 41.6 h for sensors paired with the Guardian Connect system, and 147.6 ± 40.4 h for sensors connected to the GSR. Responses to survey questions regarding sensor comfort and ease of use were favorable. The Guardian Sensor 3 glucose sensor, whether located in abdomen or the arm, provided accurate glucose readings when compared with the YSI reference and demonstrated functional life commensurate with the intended 7-day use. ClinicalTrials.gov : NCT02246582.

O-GlcNAcase Expression is Sensitive to Changes in O-GlcNAc Homeostasis.

Science.gov (United States)

Zhang, Zhen; Tan, Ee Phie; VandenHull, Nicole J; Peterson, Kenneth R; Slawson, Chad

2014-01-01

O-linked N-acetylglucosamine (O-GlcNAc) is a post-translational modification involving an attachment of a single β-N-acetylglucosamine moiety to serine or threonine residues in nuclear and cytoplasmic proteins. Cellular O-GlcNAc levels are regulated by two enzymes: O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA), which add and remove the modification, respectively. The levels of O-GlcNAc can rapidly change in response to fluctuations in the extracellular environment; however, O-GlcNAcylation returns to a baseline level quickly after stimulus removal. This process termed O-GlcNAc homeostasis appears to be critical to the regulation of many cellular functions including cell cycle progress, stress response, and gene transcription. Disruptions in O-GlcNAc homeostasis are proposed to lead to the development of diseases, such as cancer, diabetes, and Alzheimer's disease. O-GlcNAc homeostasis is correlated with the expression of OGT and OGA. We reason that alterations in O-GlcNAc levels affect OGA and OGT transcription. We treated several human cell lines with Thiamet-G (TMG, an OGA inhibitor) to increase overall O-GlcNAc levels resulting in decreased OGT protein expression and increased OGA protein expression. OGT transcript levels slightly declined with TMG treatment, but OGA transcript levels were significantly increased. Pretreating cells with protein translation inhibitor cycloheximide did not stabilize OGT or OGA protein expression in the presence of TMG; nor did TMG stabilize OGT and OGA mRNA levels when cells were treated with RNA transcription inhibitor actinomycin D. Finally, we performed RNA Polymerase II chromatin immunoprecipitation at the OGA promoter and found that RNA Pol II occupancy at the transcription start site was lower after prolonged TMG treatment. Together, these data suggest that OGA transcription was sensitive to changes in O-GlcNAc homeostasis and was potentially regulated by O-GlcNAc.

Structure of the RBD-PRDI fragment of the antiterminator protein GlcT

International Nuclear Information System (INIS)

Himmel, Sebastian; Grosse, Christian; Wolff, Sebastian; Schwiegk, Claudia; Becker, Stefan

2012-01-01

The crystal structure of the RBD-PRDI fragment of the antiterminator protein GlcT from Bacillus subtilis has been solved at 2 Å resolution. The structure represents an inactive state of the protein. GlcT is a transcriptional antiterminator protein that is involved in regulation of glucose metabolism in Bacillus subtilis. Antiterminator proteins bind specific RNA sequences, thus preventing the formation of overlapping terminator stem-loops. The structure of a fragment (residues 3–170) comprising the RNA-binding domain (RBD) and the first regulatory domain (PRDI) of GlcT was solved at 2.0 Å resolution with one molecule in the asymmetric unit. The two domains are connected by a helical linker. Their interface is mostly constituted by hydrophobic interactions

Conditions With High Intracellular Glucose Inhibit Sensing Through Glucose Sensor Snf3 in Saccharomyces cerevisiae

DEFF Research Database (Denmark)

Karhumaa, Kaisa; Wu, B.Q.; Kielland-Brandt, Morten

2010-01-01

as for amino acids. An alternating-access model of the function of transporter-like sensors has been previously suggested based on amino acid sensing, where intracellular ligand inhibits binding of extracellular ligand. Here we studied the effect of intracellular glucose on sensing of extracellular glucose...... through the transporter-like sensor Snf3 in yeast. Sensing through Snf3 was determined by measuring degradation of Mth1 protein. High intracellular glucose concentrations were achieved by using yeast strains lacking monohexose transporters which were grown on maltose. The apparent affinity...... of extracellular glucose to Snf3 was measured for cells grown in non-fermentative medium or on maltose. The apparent affinity for glucose was lowest when the intracellular glucose concentration was high. The results conform to an alternating-access model for transporter-like sensors. J. Cell. Biochem. 110: 920...

Peptide substrate-assisted study of O-GlcNAc transferase and O-GlcNAcylation

NARCIS (Netherlands)

Shi, Jie

2018-01-01

O-GlcNAcylation is a post translational modification (PTM) that corresponds to the addition of a single β-linked N-Acetyl-D-glucosamine (GlcNAc) sugar moiety onto the hydroxyl group of serine and threonine residues in numerous proteins. The addition of O-GlcNAc to proteins is catalyzed by O-GlcNAc

O-GlcNAcase expression is sensitive to changes in O-GlcNAc homeostasis

Directory of Open Access Journals (Sweden)

ZHEN eZHANG

2014-12-01

Full Text Available O-linked N-acetylglucosamine (O-GlcNAc is a post-translational modification involving an attachment of a single β-N-acetylglucosamine moiety to serine or threonine residues in nuclear and cytoplasmic proteins. Cellular O-GlcNAc levels are regulated by two enzymes: O-GlcNAc transferase (OGT and O-GlcNAcase (OGA, which add and remove the modification respectively. The levels of O-GlcNAc can rapidly change in response to fluctuations in the extracellular environment; however, O-GlcNAcylation returns to a baseline level quickly after stimulus removal. This process termed O-GlcNAc homeostasis appears to be critical to the regulation of many cellular functions including cell cycle progress, stress response, and gene transcription. Disruptions in O-GlcNAc homeostasis are proposed to lead to the development of diseases such as cancer, diabetes, and Alzheimer’s disease. O-GlcNAc homeostasis is correlated with the expression of OGT and OGA. We reason that alterations in O-GlcNAc levels affect OGA and OGT transcription. We treated several human cell lines with Thiamet-G (TMG, an OGA inhibitor to increase overall O-GlcNAc levels resulting in decreased OGT protein expression and increased OGA protein expression. OGT transcript levels slightly declined with TMG treatment, but OGA transcript levels were significantly increased. Pretreating cells with protein translation inhibitor cycloheximide (CHX did not stabilize OGT or OGA protein expression in the presence of TMG; nor did TMG stabilize OGT and OGA mRNA levels when cells were treated with RNA transcription inhibitor actinomycin D (AMD. Finally, we performed RNA Polymerase II chromatin immunoprecipitation (ChIP at the OGA promoter and found RNA Pol II occupancy at the transcription start site (TSS was lower after prolonged TMG treatment. Together, these data suggest that OGA transcription was sensitive to changes in O-GlcNAc homeostasis and was potentially regulated by O-GlcNAc.

Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity

DEFF Research Database (Denmark)

Shi, Hao; Munk, Alexander; Nielsen, Thomas Svava

2018-01-01

-GlcNAcylation, in skeletal muscle. METHODS: We assessed O-GlcNAcylation levels in skeletal muscle from obese, type 2 diabetic people, and we characterized muscle-specific OGT knockout (mKO) mice in metabolic cages and measured energy expenditure and substrate utilization pattern using indirect calorimetry. Whole body...... of O-GlcNAc-modified proteins in obese, type 2 diabetic people compared with well-matched obese and lean controls. Muscle-specific OGT knockout mice were lean, and whole body energy expenditure and insulin sensitivity were increased in these mice, consistent with enhanced glucose uptake and elevated...

Continuous glucose monitoring in subcutaneous tissue using factory-calibrated sensors: a pilot study.

Science.gov (United States)

Hoss, Udo; Jeddi, Iman; Schulz, Mark; Budiman, Erwin; Bhogal, Claire; McGarraugh, Geoffrey

2010-08-01

Commercial continuous subcutaneous glucose monitors require in vivo calibration using capillary blood glucose tests. Feasibility of factory calibration, i.e., sensor batch characterization in vitro with no further need for in vivo calibration, requires a predictable and stable in vivo sensor sensitivity and limited inter- and intra-subject variation of the ratio of interstitial to blood glucose concentration. Twelve volunteers wore two FreeStyle Navigator (Abbott Diabetes Care, Alameda, CA) continuous glucose monitoring systems for 5 days in parallel for two consecutive sensor wears (four sensors per subject, 48 sensors total). Sensors from a prototype sensor lot with a low variability in glucose sensitivity were used for the study. Median sensor sensitivity values based on capillary blood glucose were calculated per sensor and compared for inter- and intra-subject variation. Mean absolute relative difference (MARD) calculation and error grid analysis were performed using a single calibration factor for all sensors to simulate factory calibration and compared to standard fingerstick calibration. Sensor sensitivity variation in vitro was 4.6%, which increased to 8.3% in vivo (P glucose monitoring is feasible with similar accuracy to standard fingerstick calibration. Additional data are required to confirm this result in subjects with diabetes.

Determination of Glucose Utilization Rates in Cultured Astrocytes and Neurons with [14C]deoxyglucose: Progress, Pitfalls, and Discovery of Intracellular Glucose Compartmentation.

Science.gov (United States)

Dienel, Gerald A; Cruz, Nancy F; Sokoloff, Louis; Driscoll, Bernard F

2017-01-01

2-Deoxy-D-[ 14 C]glucose ([ 14 C]DG) is commonly used to determine local glucose utilization rates (CMR glc ) in living brain and to estimate CMR glc in cultured brain cells as rates of [ 14 C]DG phosphorylation. Phosphorylation rates of [ 14 C]DG and its metabolizable fluorescent analog, 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose (2-NBDG), however, do not take into account differences in the kinetics of transport and metabolism of [ 14 C]DG or 2-NBDG and glucose in neuronal and astrocytic cells in cultures or in single cells in brain tissue, and conclusions drawn from these data may, therefore, not be correct. As a first step toward the goal of quantitative determination of CMR glc in astrocytes and neurons in cultures, the steady-state intracellular-to-extracellular concentration ratios (distribution spaces) for glucose and [ 14 C]DG were determined in cultured striatal neurons and astrocytes as functions of extracellular glucose concentration. Unexpectedly, the glucose distribution spaces rose during extreme hypoglycemia, exceeding 1.0 in astrocytes, whereas the [ 14 C]DG distribution space fell at the lowest glucose levels. Calculated CMR glc was greatly overestimated in hypoglycemic and normoglycemic cells because the intracellular glucose concentrations were too high. Determination of the distribution space for [ 14 C]glucose revealed compartmentation of intracellular glucose in astrocytes, and probably, also in neurons. A smaller metabolic pool is readily accessible to hexokinase and communicates with extracellular glucose, whereas the larger pool is sequestered from hexokinase activity. A new experimental approach using double-labeled assays with DG and glucose is suggested to avoid the limitations imposed by glucose compartmentation on metabolic assays.

Pivotal Role of O-GlcNAc Modification in Cold-Induced Thermogenesis by Brown Adipose Tissue Through Mitochondrial Biogenesis.

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Ohashi, Natsuko; Morino, Katsutaro; Ida, Shogo; Sekine, Osamu; Lemecha, Mengistu; Kume, Shinji; Park, Shi-Young; Choi, Cheol Soo; Ugi, Satoshi; Maegawa, Hiroshi

2017-09-01

Adipose tissues considerably influence metabolic homeostasis, and both white (WAT) and brown (BAT) adipose tissue play significant roles in lipid and glucose metabolism. O -linked N -acetylglucosamine ( O -GlcNAc) modification is characterized by the addition of N -acetylglucosamine to various proteins by O -GlcNAc transferase (Ogt), subsequently modulating various cellular processes. However, little is known about the role of O -GlcNAc modification in adipose tissues. Here, we report the critical role of O -GlcNAc modification in cold-induced thermogenesis. Deletion of Ogt in WAT and BAT using adiponectin promoter-driven Cre recombinase resulted in severe cold intolerance with decreased uncoupling protein 1 (Ucp1) expression. Furthermore, Ogt deletion led to decreased mitochondrial protein expression in conjunction with decreased peroxisome proliferator-activated receptor γ coactivator 1-α protein expression. This phenotype was further confirmed by deletion of Ogt in BAT using Ucp1 promoter-driven Cre recombinase, suggesting that O -GlcNAc modification in BAT is responsible for cold-induced thermogenesis. Hypothermia was significant under fasting conditions. This effect was mitigated after normal diet consumption but not after consumption of a fatty acid-rich ketogenic diet lacking carbohydrates, suggesting impaired diet-induced thermogenesis, particularly by fat. In conclusion, O -GlcNAc modification is essential for cold-induced thermogenesis and mitochondrial biogenesis in BAT. Glucose flux into BAT may be a signal to maintain BAT physiological responses. © 2017 by the American Diabetes Association.

O-Linked β-N-acetylglucosamine (O-GlcNAc) modification: a new pathway to decode pathogenesis of diabetic retinopathy.

Science.gov (United States)

Gurel, Zafer; Sheibani, Nader

2018-01-31

The incidence of diabetes continues to rise among all ages and ethnic groups worldwide. Diabetic retinopathy (DR) is a complication of diabetes that affects the retinal neurovasculature causing serious vision problems, including blindness. Its pathogenesis and severity is directly linked to the chronic exposure to high glucose conditions. No treatments are currently available to stop the development and progression of DR. To develop new and effective therapeutic approaches, it is critical to better understand how hyperglycemia contributes to the pathogenesis of DR at the cellular and molecular levels. We propose alterations in O-GlcNAc modification of target proteins during diabetes contribute to the development and progression of DR. The O-GlcNAc modification is regulated through hexosamine biosynthetic pathway. We showed this pathway is differentially activated in various retinal vascular cells under high glucose conditions perhaps due to their selective metabolic activity. O-GlcNAc modification can alter protein stability, activity, interactions, and localization. By targeting the same amino acid residues (serine and threonine) as phosphorylation, O-GlcNAc modification can either compete or cooperate with phosphorylation. Here we will summarize the effects of hyperglycemia-induced O-GlcNAc modification on the retinal neurovasculature in a cell-specific manner, providing new insight into the role of O-GlcNAc modification in early loss of retinal pericytes and the pathogenesis of DR. © 2018 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Elevated O-GlcNAcylation stabilizes FOXM1 by its reduced degradation through GSK-3β inactivation in a human gastric carcinoma cell line, MKN45 cells.

Science.gov (United States)

Inoue, Yosuke; Moriwaki, Kazumasa; Ueda, Yasuhiro; Takeuchi, Toshihisa; Higuchi, Kazuhide; Asahi, Michio

2018-01-08

O-GlcNAcylation is a dynamic post-translational modification of cytonuclear proteins for intracellular signaling. Elevated O-GlcNAcylation is a general feature of cancer and contributes to cancer progression, and recent studies indicate the contribution to increasing incidence of various types of cancer in diabetic patients. However, the role of O-GlcNAcylation in tumor progression is not fully elucidated. Forkhead box M1 (FOXM1), a master mitotic transcription factor, has been implicated in all major hallmarks of cancer, and is wildly expressed in solid tumors. Given that FOXM1 expression was reported to be elevated in gastric cancer, we examined the effect of high glucose or an inhibitor of O-GlcNAc hydrolase, Thiamet G (TMG), on FOXM1 protein expression in a human gastric cancer cell line, MKN45 cells, and confirmed that FOXM1 protein level and the cell proliferation were upregulated. To investigate the molecular mechanisms by which FOXM1 protein expression is regulated by O-GlcNAcylation, the effect of high glucose and TMG on FOXM1 ubiquitination was examined in MKN45 cells. As a result, the ubiquitination and degradation of FOXM1 protein were both suppressed by high glucose and TMG treatment. However, the O-GlcNAcylation was not detected on FOXM1 but not on GSK-3β. High glucose and TMG treatment increased phospho-serine 9 GSK-3β, an inactive form, and the degradation of FOXM1 protein was suppressed by treatment of GSK-3β inhibitors in MKN45 cells. Taken together, we suggest that high glucose and elevated O-GlcNAcylation stabilize FOXM1 protein by its reduced degradation via GSK-3β inactivation in MKN45 cells, suggesting that the higher risk of gastric cancer in diabetic patients could be partially due to O-GlcNAcylation-mediated FOXM1 stabilization. Copyright © 2017 Elsevier Inc. All rights reserved.

Fabrication of Amperometric Glucose Sensor Using Glucose Oxidase-Cellulose Nanofiber Aqueous Solution.

Science.gov (United States)

Yasuzawa, Mikito; Omura, Yuya; Hiura, Kentaro; Li, Jiang; Fuchiwaki, Yusuke; Tanaka, Masato

2015-01-01

Cellulose nanofiber aqueous solution, which remained virtually transparent for more than one week, was prepared by using the clear upper layer of diluted cellulose nanofiber solution produced by wet jet milling. Glucose oxidase (GOx) was easily dissolved in this solution and GOx-immobilized electrode was easily fabricated by simple repetitious drops of GOx-cellulose solution on the surface of a platinum-iridium electrode. Glucose sensor properties of the obtained electrodes were examined in phosphate buffer solution of pH 7.4 at 40°C. The obtained electrode provided a glucose sensor response with significantly high response speed and good linear relationship between glucose concentration and response current. After an initial decrease of response sensitivity for a few days, relatively constant sensitivity was obtained for about 20 days. Nevertheless, the influence of electroactive compounds such as ascorbic acid, uric acid and acetoaminophen were not negletable.

A role for N-acetylglucosamine as a nutrient sensor and mediator of insulin resistance.

Science.gov (United States)

Wells, L; Vosseller, K; Hart, G W

2003-02-01

The ability to regulate energy balance at both the cellular and whole body level is an essential process of life. As western society has shifted to a higher caloric diet and more sedentary lifestyle, the incidence of type 2 diabetes (non-insulin-dependent diabetes mellitus) has increased to epidemic proportions. Thus, type 2 diabetes has been described as a disease of 'chronic overnutrition'. There are abundant data to support the relationship between nutrient availability and insulin action. However, there have been multiple hypotheses and debates as to the mechanism by which nutrient availability modulates insulin signaling and how excess nutrients lead to insulin resistance. One well-established pathway for nutrient sensing is the hexosamine biosynthetic pathway (HSP), which produces the acetylated aminosugar nucleotide uridine 5'-diphospho-N-acetylglucosamine (UDP-Glc-NAc) as its end product. Since UDP-GlcNAc is the donor substrate for modification of nucleocytoplasmic proteins at serine and threonine residues with N-acetylglucosamine (O-GlcNAc), the possibility of this posttranslational modification serving as the nutrient sensor has been proposed. We have recently directly tested this model in adipocytes by examining the effect of elevated levels of O-GlcNAc on insulin-stimulated glucose uptake. In this review, we summarize the existing work that implicates the HSP and O-GlcNAc modification as nutrient sensors and regulators of insulin signaling.

A Highly Sensitive Electrochemical Glucose Sensor By Nickel-Epoxy Electrode With Non-Enzymatic Sensor

Directory of Open Access Journals (Sweden)

Riyanto Riyanto

2016-03-01

Full Text Available The preparation of new sensor for glucose was based on the fact that glucose can be determined by non-enzymatic glucose oxidase. The Ni metals (99.98% purity, 0.5 mm thick, Aldrich Chemical Company was used to prepare Ni-Epoxy electrode. The Ni-epoxy electrodes were prepared in square cut of 1 cm and 1 mm by length and wide respectively. The Ni metal electrodes were connected to silver wire with silver conducting paint prior covered with epoxy gum. The prepared of nickel-epoxy modified electrode showed outstanding electro catalytic activity toward the oxidation of glucose in alkaline solution. The result from this research are correlation of determination using Nickel-Epoxyelectrode for electroanalysis of glucose in NaOH was R2 = 0.9984. LOQ, LOD and recovery of the Nickel-Epoxy electrode towards glucose were found to be 4.4 μM, 1.48 μM and 98.19%, respectively. The Nickel-Epoxy wire based electrochemical glucose sensor demonstrates good sensitivity, wide linear range, outstanding detection limit, attractive selectivity, good reproducibility, high stability as well as prominent feasibility use of non-enzymatic sensor for monitoring glucose in human urine owing to its advantages of low cost, simple preparation and excellent properties for glucose detection.

The global regulatory system Csr senses glucose through the phosphoenolpyruvate: carbohydrate phosphotransferase system.

Science.gov (United States)

Pérez-Morales, Deyanira; Bustamante, Víctor H

2016-02-01

A novel connection between two regulatory systems controlling crucial biological processes in bacteria, the carbon storage regulator (Csr) system and the glucose-specific phosphotransferase system (PTS), is reported by Leng et al. in this issue. This involves the interaction of unphosphorylated EIIA(Glc), a component of the glucose-specific PTS, with the CsrD protein, which accelerates the decay of the CsrB and CsrC small RNAs via RNase E in Escherichia coli. As unphosphorylated EIIA(G) (lc) is generated in the presence of glucose, the PTS thus acts as a sensor of glucose for the Csr system. Interestingly, another pathway can operate for communication between the Csr system and the glucose-specific PTS. The absence of glucose generates phosphorylated EIIA(Glc) , which activates the enzyme adenylate cyclase to produce cyclic adenosine monophosphate (cAMP) that, in turn, binds to the regulator cAMP receptor protein (CRP). Leng et al. show that the complex cAMP-CRP modestly reduces CsrB decay independently of CsrD. On the other hand, a previous study indicates that the complex cAMP-CRP positively regulates the transcription of CsrB and CsrC in Salmonella enterica. Therefore, EIIA(G) (lc) could work as a molecular switch that regulates the activity of the Csr system, in response to its phosphorylation state determined by the presence or absence of glucose, in order to control gene expression. © 2015 John Wiley & Sons Ltd.

Evanescent Wave Absorption Based Fiber Sensor for Measuring Glucose Solution Concentration

Science.gov (United States)

Marzuki, Ahmad; Candra Pratiwi, Arni; Suryanti, Venty

2018-03-01

An optical fiber sensor based on evanescent wave absorption designed for measuring glucose solution consentration was proposed. The sensor was made to detect absorbance of various wavelength in the glucose solution. The sensing element was fabricated by side polishing of multimode polymer optical fiber to form a D-shape. The sensing element was immersed in different concentration of glucoce solution. As light propagated through the optical fiber, the evanescent wave interacted with the glucose solution. Light was absorbed by the glucose solution. The larger concentration the glucose solution has, the more the evanescent wave was absorbed in particular wavelenght. Here in this paper, light absorbtion as function of glucose concentration was measured as function of wavelength (the color of LED). We have shown that the proposed sensor can demonstrated an increase of light absorption as function of glucose concentration.

A Loose Relationship: Incomplete H+/Sugar Coupling in the MFS Sugar Transporter GlcP.

Science.gov (United States)

Bazzone, Andre; Zabadne, Annas J; Salisowski, Anastasia; Madej, M Gregor; Fendler, Klaus

2017-12-19

The glucose transporter from Staphylococcus epidermidis, GlcP Se , is a homolog of the human GLUT sugar transporters of the major facilitator superfamily. Together with the xylose transporter from Escherichia coli, XylE Ec , the other prominent prokaryotic GLUT homolog, GlcP Se , is equipped with a conserved proton-binding site arguing for an electrogenic transport mode. However, the electrophysiological analysis of GlcP Se presented here reveals important differences between the two GLUT homologs. GlcP Se , unlike XylE Ec , does not perform steady-state electrogenic transport at symmetrical pH conditions. Furthermore, when a pH gradient is applied, partially uncoupled transport modes can be generated. In contrast to other bacterial sugar transporters analyzed so far, in GlcP Se sugar binding, translocation and release are also accomplished by the deprotonated transporter. Based on these experimental results, we conclude that coupling of sugar and H + transport is incomplete in GlcP Se . To verify the viability of the observed partially coupled GlcP Se transport modes, we propose a universal eight-state kinetic model in which any degree of coupling is realized and H + /sugar symport represents only a specific instance. Furthermore, using sequence comparison with strictly coupled XylE Ec and similar sugar transporters, we identify an additional charged residue that may be essential for effective H + /sugar symport. Copyright © 2017 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Glucose Sensor Using U-Shaped Optical Fiber Probe with Gold Nanoparticles and Glucose Oxidase.

Science.gov (United States)

Chen, Kuan-Chieh; Li, Yu-Le; Wu, Chao-Wei; Chiang, Chia-Chin

2018-04-16

In this study, we proposed a U-shaped optical fiber probe fabricated using a flame heating method. The probe was packaged in glass tube to reduce human factors during experimental testing of the probe as a glucose sensor. The U-shaped fiber probe was found to have high sensitivity in detecting the very small molecule. When the sensor was dipped in solutions with different refractive indexes, its wavelength or transmission loss changed. We used electrostatic self-assembly to bond gold nanoparticles and glucose oxidase (GOD) onto the sensor’s surface. The results over five cycles of the experiment showed that, as the glucose concentration increased, the refractive index of the sensor decreased and its spectrum wavelength shifted. The best wavelength sensitivity was 2.899 nm/%, and the linearity was 0.9771. The best transmission loss sensitivity was 5.101 dB/%, and the linearity was 0.9734. Therefore, the proposed U-shaped optical fiber probe with gold nanoparticles and GOD has good potential for use as a blood sugar sensor in the future.

Development of a nanowire based titanium needle probe sensor for glucose monitoring

Science.gov (United States)

Deshpande, Devesh C.

The need for continuous monitoring of various physiological functions such as blood glucose levels, neural functions and cholesterol levels has fostered the research and development of various schemes of biosensors to sense and help control the respective function. The needs of patients for sensors with minimal discomfort, longer life and better performance have necessitated the development towards smaller and more efficient sensors. In addition, the need for higher functionality from smaller sensors has led to the development of sensors with multiple electrodes, each electrode capable of sensing a different body function. Such multi-electrode sensors need to be fabricated using micro-fabrication processes in order to achieve precise control over the size, shape and placement of the electrodes. Multielectrode sensors fabricated using silicon and polymers have been demonstrated. One physiological function that attracts widespread interest is continuous glucose monitoring in our blood, since Diabetes affects millions of people all over the world. Significant deviations of blood glucose levels from the normal levels of 4-8 mM can cause fainting, coma and damage to the eyes, kidneys, nerves and blood vessels. For chronic patients, continuous monitoring of glucose levels is essential for accurate and timely treatment. A few continuous monitoring sensors are available in the market, but they have problems and cannot replace the strip type one-time glucose monitoring systems as yet. To address this need, large scale research efforts have been targeted towards continuous monitoring. The demand for higher accuracy and sensitivity has motivated researchers to evaluate the use of nanostructures in sensing. The large surface area-to-volume ratio of such structures could enable further miniaturization and push the detection limits, potentially enabling even single molecule detection. This research involved the development of a biocompatible titanium needle probe sensor for

Disruption of O-GlcNAc cycling in C. elegans perturbs Nucleotide Sugar pools and Complex Glycans

Directory of Open Access Journals (Sweden)

Salil K Ghosh

2014-11-01

Full Text Available The carbohydrate modification of serine and threonine residues with O-linked beta-N-acetylglucosamine (O-GlcNAc is ubiquitous and governs cellular processes ranging from cell signaling to apoptosis. The O-GlcNAc modification along with other carbohydrate modifications, including N-linked and O-linked glycans, glycolipids, and sugar polymers, all require the use of the nucleotide sugar UDP-GlcNAc, the end product of the hexosamine biosynthetic pathway. In this paper, we describe the biochemical consequences resulting from perturbation of the O-GlcNAc pathway in C. elegans lacking O-GlcNAc transferase and O-GlcNAcase activities. In ogt-1 null animals, steady-state levels of UDP-GlcNAc/UDP-GalNAc and UDP-glucose were substantially elevated. Transcripts of genes encoding for key members in the Hexosamine Biosynthetic Pathway (gfat-2, gna-2, C36A4.4 and trehalose metabolism (tre-1, tre-2, and tps-2 were elevated in ogt-1 null animals. While there is no evidence to suggest changes in the profile of N-linked glycans in the ogt-1 and oga-1 mutants, glycans insensitive to PNGase digestion (including O-linked glycans, glycolipids, and glycopolymers were altered in these strains. Our data supports that changes in O-GlcNAcylation alters nucleotide sugar production, overall glycan composition, and transcription of genes encoding glycan processing enzymes. These data along with our previous findings that disruption in O-GlcNAc cycling alters macronutrient storage underscores the noteworthy influence this posttranslational modification plays in nutrient sensing.

Highly Selective and Sensitive Self-Powered Glucose Sensor Based on Capacitor Circuit.

Science.gov (United States)

Slaughter, Gymama; Kulkarni, Tanmay

2017-05-03

Enzymatic glucose biosensors are being developed to incorporate nanoscale materials with the biological recognition elements to assist in the rapid and sensitive detection of glucose. Here we present a highly sensitive and selective glucose sensor based on capacitor circuit that is capable of selectively sensing glucose while simultaneously powering a small microelectronic device. Multi-walled carbon nanotubes (MWCNTs) is chemically modified with pyrroloquinoline quinone glucose dehydrogenase (PQQ-GDH) and bilirubin oxidase (BOD) at anode and cathode, respectively, in the biofuel cell arrangement. The input voltage (as low as 0.25 V) from the biofuel cell is converted to a stepped-up power and charged to the capacitor to the voltage of 1.8 V. The frequency of the charge/discharge cycle of the capacitor corresponded to the oxidation of glucose. The biofuel cell structure-based glucose sensor synergizes the advantages of both the glucose biosensor and biofuel cell. In addition, this glucose sensor favored a very high selectivity towards glucose in the presence of competing and non-competing analytes. It exhibited unprecedented sensitivity of 37.66 Hz/mM.cm 2 and a linear range of 1 to 20 mM. This innovative self-powered glucose sensor opens new doors for implementation of biofuel cells and capacitor circuits for medical diagnosis and powering therapeutic devices.

77 FR 30016 - Clinical Study Design and Performance of Hospital Glucose Sensors

Science.gov (United States)

2012-05-21

...] Clinical Study Design and Performance of Hospital Glucose Sensors AGENCY: Food and Drug Administration, HHS... Sensors.'' The purpose of this public meeting is to discuss clinical study design considerations and performance metrics for innovative glucose sensors intended to be used in hospital point of care settings...

A highly sensitive electrochemical glucose sensor structuring with nickel hydroxide and enzyme glucose oxidase

International Nuclear Information System (INIS)

Mathew, Manjusha; Sandhyarani, N.

2013-01-01

Graphical abstract: A combination of Ni 2+ /Ni 3+ redox couple and glucose oxidase has successfully been exploited for the realization of a highly sensitive glucose sensor for the first time. -- Highlights: • A multilayered glucose biosensor with enhanced sensitivity was fabricated. • Combination of Ni 2+ /Ni 3+ redox couple and glucose oxidase has been exploited for the first time. • Exhibits a lower detection limit of 100 nM with a high sensitivity of 16,840 μA mM −1 cm −2 . • The surface shows a low Michaelis–Menten constant value of 2.4 μM. • Detailed mechanism of sensing was proposed and justified. -- Abstract: A multilayered glucose biosensor with enhanced electron transport was fabricated via the sequential electrodeposition of chitosan gold nanocomposite (CGNC) and nickel hydroxide (Ni(OH) 2 ) on a bare gold electrode and subsequent immobilization of glucose oxidase. A thin film of Ni(OH) 2 deposited on CGNC modified gold electrode serves as an electrochemical redox probe as well as a matrix for the immobilization of glucose oxidase retaining its activity. Electron transport property of CGNC has been exploited to enhance the electron transport between the analyte and electrode. Electrochemical characteristics of the biosensor were studied by cyclic voltammetry and chronoamperometry. Under optimal conditions the biosensor exhibits a linear range from 1 μM to 100 μM with a limit of detection (lod) down to 100 nM. The sensor shows a low Michaelis-Menten constant value of 2.4 μM indicates the high affinity of enzyme to the analyte points to the retained activity of enzyme after immobilization. The present glucose sensor with the high selectivity, sensitivity and stability is promising for practical clinical applications

Addressing glucose sensitivity measured by F-18 FDG PET in lung cancers for radiation treatment planning and monitoring

International Nuclear Information System (INIS)

Wong, Ching-yee Oliver; Thie, Joseph; Gaskill, Marianne; Kestin, Larry; Yan Di; Cheng, Vincent; Nagle, Conrad

2006-01-01

Purpose: To address glucose sensitivity in lung cancers before and after radiation treatment (Tx). Methods and Materials: Twelve patients were each studied with two pre-Tx positron emission tomography (PET) scans and 3 patients each with one post-Tx PET scan, with glucose concentration [Glc] and maximum standard uptake value (SUV) recorded. The pre-Tx glucose sensitivity, g from SUV 1 /SUV 2 = {[Glc] 1 /[Glc] 2 } g and Tx index, τ from SUV post-Tx /SUV pre-Tx = {[Glc] post-Tx /[Glc] pre-Tx } τ was calculated by linear regression. Pre-Tx SUVs were corrected to post-Tx Glc with g (SUV' pre-Tx ) for a pure Tx effect, R ln(SUV post-Tx /SUV' pre-Tx ). Results: There were no significant differences in SUV but [Glc] were different (96.4 ± 10.9 vs. 88.3 ± 10.5, p = 0.015) between two pre-Tx PET scans. Linear regression yielded g -0.79 and τ = -1.78 to -2.41 (p < 0.0005 in all). The %ΔSUV after Tx for 3 patients without vs. with g correction were different by -12%, 0%, and + 7%, suggesting varying effects from glucose. R values were also different and mean R (-0.81 ± 0.38) was significantly different from zero (p = 0.03), consistent with successful Tx as confirmed by clinico-radiologic follow-up. Conclusions: The extra dimension of glucose sensitivity, g besides SUV incorporated in the combined Tx-derived τ may be a useful global Tx evaluation index even with differing [Glc

POF based glucose sensor incorporating grating wavelength filters

DEFF Research Database (Denmark)

Hassan, Hafeez Ul; Aasmul, Søren; Bang, Ole

2014-01-01

AND RESEARCH IN POLYMER OPTICAL DEVICES; TRIPOD. Within the domain of TRIPOD, research is conducted on "Plastic Optical Fiber based Glucose Sensors Incorporating Grating Wavelength Filters". Research will be focused to optimized fiber tips for better coupling efficiency, reducing the response time of sensor...

A low-potential, H{sub 2}O{sub 2}-assisted electrodeposition of cobalt oxide/hydroxide nanostructures onto vertically-aligned multi-walled carbon nanotube arrays for glucose sensing

Energy Technology Data Exchange (ETDEWEB)

Yang Jiang [Food and Bioprocess Engineering laboratory, Department of Biological Systems Engineering, University of Wisconsin-Madison, 460 Henry Mall, Madison, WI 53706 (United States); Zhang Weide [Nanoscience Research Center, School of Chemistry and Chemical Engineering, South China University of Technology, Guangzhou 510630 (China); Gunasekaran, Sundaram, E-mail: guna@wisc.edu [Food and Bioprocess Engineering laboratory, Department of Biological Systems Engineering, University of Wisconsin-Madison, 460 Henry Mall, Madison, WI 53706 (United States)

2011-06-30

Highlights: > We successfully synthesized CoOx.nH{sub 2}O-MWCNTs nanocomposites using a cathodic electrochemical reduction of H{sub 2}O{sub 2} to deposit cobalt oxide/hydroxide nanostructures onto vertically well-aligned MWCNTs arrays. > This is an enzyme-free sensor. > Under optimal detection conditions, the sensor showed a good-enough sensitivity of 162.8 {mu}A mM{sup -1} cm{sup -2}, a low detection limit of 2.0 {mu}M (S/N = 3) and a fast response of less than 4 s within the linear range of up to 4.5 mM. > Other advantages of the sensor for Glc measurements include high insensitivity to common interferences, long-term stability, reproducibility and resistance to chloride poisoning without additional outer membrance like Nafion. Therefor it is useful for routine Glc analysis. > The novel nanocomposite material with good mechanical strength and high conductivity can be planted into microchannels to conduct sophisticated lab-on-a-chip Glc detection. - Abstract: A novel nanocomposite was synthesized using a cathodic, low-potential, electrochemical reduction of H{sub 2}O{sub 2} to homogeneously deposit cobalt oxide/hydroxide (denoted as CoOx.nH{sub 2}O) nanostructures onto vertically well-aligned multi-walled carbon nanotube arrays (MWCNTs), while the MWCNTs were prepared by catalytic chemical vapor deposition (CVD) on a tantalum (Ta) substrate. The CoOx.nH{sub 2}O-MWCNTs nanocomposite exhibits much higher electrocatalytic activity towards glucose (Glc) after modification with CoOx.nH{sub 2}O than before. This non-enzymatic Glc sensor has a high sensitivity (162.8 {mu}A mM{sup -1} cm{sup -2}), fast response time (<4 s) and low detection limit (2.0 {mu}M at signal/noise ratio = 3), and a linear dynamic range up to 4.5 mM. The sensor output is stable over 30 days and unaffected by common interferents that co-exist with Glc in analytical samples; it is also resistant to chloride poisoning. These features make the CoOx.nH{sub 2}O-MWCNTs nanocomposite a promising electrode

Porous, Dexamethasone-loaded polyurethane coatings extend performance window of implantable glucose sensors in vivo.

Science.gov (United States)

Vallejo-Heligon, Suzana G; Brown, Nga L; Reichert, William M; Klitzman, Bruce

2016-01-01

Continuous glucose sensors offer the promise of tight glycemic control for insulin dependent diabetics; however, utilization of such systems has been hindered by issues of tissue compatibility. Here we report on the in vivo performance of implanted glucose sensors coated with Dexamethasone-loaded (Dex-loaded) porous coatings employed to mediate the tissue-sensor interface. Two animal studies were conducted to (1) characterize the tissue modifying effects of the porous Dex-loaded coatings deployed on sensor surrogate implants and (2) investigate the effects of the same coatings on the in vivo performance of Medtronic MiniMed SOF-SENSOR™ glucose sensors. The tissue response to implants was evaluated by quantifying macrophage infiltration, blood vessel formation, and collagen density around implants. Sensor function was assessed by measuring changes in sensor sensitivity and time lag, calculating the Mean Absolute Relative Difference (MARD) for each sensor treatment, and performing functional glucose challenge test at relevant time points. Implants treated with porous Dex-loaded coatings diminished inflammation and enhanced vascularization of the tissue surrounding the implants. Functional sensors with Dex-loaded porous coatings showed enhanced sensor sensitivity over a 21-day period when compared to controls. Enhanced sensor sensitivity was accompanied with an increase in sensor signal lag and MARD score. These results indicate that Dex-loaded porous coatings were able to elicit an attenuated tissue response, and that such tissue microenvironment could be conducive towards extending the performance window of glucose sensors in vivo. In the present article, a coating to extend the functionality of implantable glucose sensors in vivo was developed. Our study showed that the delivery of an anti-inflammatory agent with the presentation of micro-sized topographical cues from coatings may lead to improved long-term glucose sensor function in vivo. We believe that

Toward an injectable continuous osmotic glucose sensor.

Science.gov (United States)

Johannessen, Erik; Krushinitskaya, Olga; Sokolov, Andrey; Philipp, Häfliger; Hoogerwerf, Arno; Hinderling, Christian; Kautio, Kari; Lenkkeri, Jaakko; Strömmer, Esko; Kondratyev, Vasily; Tønnessen, Tor Inge; Mollnes, Tom Eirik; Jakobsen, Henrik; Zimmer, Even; Akselsen, Bengt

2010-07-01

The growing pandemic of diabetes mellitus places a stringent social and economic burden on the society. A tight glycemic control circumvents the detrimental effects, but the prerogative is the development of new more effective tools capable of longterm tracking of blood glucose (BG) in vivo. Such discontinuous sensor technologies will benefit from an unprecedented marked potential as well as reducing the current life expectancy gap of eight years as part of a therapeutic regime. A sensor technology based on osmotic pressure incorporates a reversible competitive affinity assay performing glucose-specific recognition. An absolute change in particles generates a pressure that is proportional to the glucose concentration. An integrated pressure transducer and components developed from the silicon micro- and nanofabrication industry translate this pressure into BG data. An in vitro model based on a 3.6 x 8.7 mm large pill-shaped implant is equipped with a nanoporous membrane holding 4-6 nm large pores. The affinity assay offers a dynamic range of 36-720 mg/dl with a resolution of +/-16 mg/dl. An integrated 1 x 1 mm(2) large control chip samples the sensor signals for data processing and transmission back to the reader at a total power consumption of 76 microW. Current studies have demonstrated the design, layout, and performance of a prototype osmotic sensor in vitro using an affinity assay solution for up to four weeks. The small physical size conforms to an injectable device, forming the basis of a conceptual monitor that offers a tight glycemic control of BG. 2010 Diabetes Technology Society.

A Conserved Splicing Silencer Dynamically Regulates O-GlcNAc Transferase Intron Retention and O-GlcNAc Homeostasis

Directory of Open Access Journals (Sweden)

Sung-Kyun Park

2017-08-01

Full Text Available Modification of nucleocytoplasmic proteins with O-GlcNAc regulates a wide variety of cellular processes and has been linked to human diseases. The enzymes O-GlcNAc transferase (OGT and O-GlcNAcase (OGA add and remove O-GlcNAc, but the mechanisms regulating their expression remain unclear. Here, we demonstrate that retention of the fourth intron of OGT is regulated in response to O-GlcNAc levels. We further define a conserved intronic splicing silencer (ISS that is necessary for OGT intron retention. Deletion of the ISS in colon cancer cells leads to increases in OGT, but O-GlcNAc homeostasis is maintained by concomitant increases in OGA protein. However, the ISS-deleted cells are hypersensitive to OGA inhibition in culture and in soft agar. Moreover, growth of xenograft tumors from ISS-deleted cells is compromised in mice treated with an OGA inhibitor. Thus, ISS-mediated regulation of OGT intron retention is a key component in OGT expression and maintaining O-GlcNAc homeostasis.

A Robust, Enzyme-Free Glucose Sensor Based on Lysine-Assisted CuO Nanostructures

Directory of Open Access Journals (Sweden)

Qurrat-ul-Ain Baloach

2016-11-01

Full Text Available The production of a nanomaterial with enhanced and desirable electrocatalytic properties is of prime importance, and the commercialization of devices containing these materials is a challenging task. In this study, unique cupric oxide (CuO nanostructures were synthesized using lysine as a soft template for the evolution of morphology via a rapid and boiled hydrothermal method. The morphology and structure of the synthesized CuO nanomaterial were characterized using scanning electron microscopy (SEM and X-ray diffraction (XRD, respectively. The prepared CuO nanostructures showed high potential for use in the electrocatalytic oxidation of glucose in an alkaline medium. The proposed enzyme-free glucose sensor demonstrated a robust response to glucose with a wide linear range and high sensitivity, selectivity, stability, and reproducibility. To explore its practical feasibility, the glucose content of serum samples was successfully determined using the enzyme-free sensor. An analytical recovery method was used to measure the actual glucose from the serum samples, and the results were satisfactory. Moreover, the presented glucose sensor has high chemical stability and can be reused for repetitive measurements. This study introduces an enzyme-free glucose sensor as an alternative tool for clinical glucose quantification.

An alpha-glucose-1-phosphate phosphodiesterase is present in rat liver cytosol

International Nuclear Information System (INIS)

Srisomsap, C.; Richardson, K.L.; Jay, J.C.; Marchase, R.B.

1989-01-01

UDP-glucose:glycoprotein glucose-1-phosphotransferase (Glc-phosphotransferase) catalyzes the transfer of alpha-Glc-1-P from UDP-Glc to mannose residues on acceptor glycoproteins. The predominant acceptor for this transfer in both mammalian cells and Paramecium is a cytoplasmic glycoprotein of 62-63 kDa. When cytoplasmic proteins from rat liver were fractionated by preparative isoelectric focusing following incubation of a liver homogenate with the 35S-labeled phosphorothioate analogue of UDP-Glc ([beta-35S]UDP-Glc), the acceptor was found to have a pI of about 6.0. This fraction, when not labeled prior to the focusing, became very heavily labeled when mixed with [beta-35S]. UDP-Glc and intact liver microsomes, a rich source of the Glc-phosphotransferase. In addition, it was observed that the isoelectric fractions of the cytosol having pI values of 2-3.2 contained a degradative activity, alpha-Glc-1-P phosphodiesterase, that was capable of removing alpha-Glc-1-P, monitored through radioactive labeling both in the sugar and the phosphate, as an intact unit from the 62-kDa acceptor. Identification of the product of this cleavage was substantiated by its partial transformation to UDP-Glc in the presence of UTP and UDP-Glc pyrophosphorylase. The alpha-Glc-1-P phosphodiesterase had a pH optimum of 7.5 and was not effectively inhibited by any of the potential biochemical inhibitors that were tested. Specificity for the Glc-alpha-1-P-6-Man diester was suggested by the diesterase's inability to degrade UDP-Glc or glucosylphosphoryldolichol. This enzyme may be important in the regulation of secretion since the alpha-Glc-1-P present on the 62-kDa phosphoglycoprotein appears to be removed and then rapidly replaced in response to secretagogue

Polymer Optical Fiber Compound Parabolic Concentrator fiber tip based glucose sensor: In-Vitro Testing

DEFF Research Database (Denmark)

Hassan, Hafeez Ul; Janting, Jakob; Aasmul, Soren

2016-01-01

We present in-vitro sensing of glucose using a newly developed efficient optical fiber glucose sensor based on a Compound Parabolic Concentrator (CPC) tipped polymer optical fiber (POF). A batch of 9 CPC tipped POF sensors with a 35 mm fiber length is shown to have an enhanced fluorescence pickup...... efficiency with an average increment factor of 1.7 as compared to standard POF sensors with a plane cut fiber tip. Invitro measurements for two glucose concentrations (40 and 400 mg/dL) confirm that the CPC tipped sensors efficiently can detect both glucose concentrations. it sets the footnote at the bottom...

Rewiring the Glucose Transportation and Central Metabolic Pathways for Overproduction of N-Acetylglucosamine in Bacillus subtilis.

Science.gov (United States)

Gu, Yang; Deng, Jieying; Liu, Yanfeng; Li, Jianghua; Shin, Hyun-Dong; Du, Guocheng; Chen, Jian; Liu, Long

2017-10-01

N-acetylglucosamine (GlcNAc) is an important amino sugar extensively used in the healthcare field. In a previous study, the recombinant Bacillus subtilis strain BSGN6-P xylA -glmS-pP43NMK-GNA1 (BN0-GNA1) had been constructed for microbial production of GlcNAc by pathway design and modular optimization. Here, the production of GlcNAc is further improved by rewiring both the glucose transportation and central metabolic pathways. First, the phosphotransferase system (PTS) is blocked by deletion of three genes, yyzE (encoding the PTS system transporter subunit IIA YyzE), ypqE (encoding the PTS system transporter subunit IIA YpqE), and ptsG (encoding the PTS system glucose-specific EIICBA component), resulting in 47.6% increase in the GlcNAc titer (from 6.5 ± 0.25 to 9.6 ± 0.16 g L -1 ) in shake flasks. Then, reinforcement of the expression of the glcP and glcK genes and optimization of glucose facilitator proteins are performed to promote glucose import and phosphorylation. Next, the competitive pathways for GlcNAc synthesis, namely glycolysis, peptidoglycan synthesis pathway, pentose phosphate pathway, and tricarboxylic acid cycle, are repressed by initiation codon-optimization strategies, and the GlcNAc titer in shake flasks is improved from 10.8 ± 0.25 to 13.2 ± 0.31 g L -1 . Finally, the GlcNAc titer is further increased to 42.1 ± 1.1 g L -1 in a 3-L fed-batch bioreactor, which is 1.72-fold that of the original strain, BN0-GNA1. This study shows considerably enhanced GlcNAc production, and the metabolic engineering strategy described here will be useful for engineering other prokaryotic microorganisms for the production of GlcNAc and related molecules. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Micro-Electromechanical Affinity Sensor for the Monitoring of Glucose in Bioprocess Media

Directory of Open Access Journals (Sweden)

Lorenz Theuer

2017-06-01

Full Text Available An affinity-viscometry-based micro-sensor probe for continuous glucose monitoring was investigated with respect to its suitability for bioprocesses. The sensor operates with glucose and dextran competing as binding partner for concanavalin A, while the viscosity of the assay scales with glucose concentration. Changes in viscosity are determined with a micro-electromechanical system (MEMS in the measurement cavity of the sensor probe. The study aimed to elucidate the interactions between the assay and a typical phosphate buffered bacterial cultivation medium. It turned out that contact with the medium resulted in a significant long-lasting drift of the assay’s viscosity at zero glucose concentration. Adding glucose to the medium lowers the drift by a factor of eight. The cglc values measured off-line with the glucose sensor for monitoring of a bacterial cultivation were similar to the measurements with an enzymatic assay with a difference of less than ±0.15 g·L−1. We propose that lectin agglomeration, the electro-viscous effect, and constitutional changes of concanavalin A due to exchanges of the incorporated metal ions may account for the observed viscosity increase. The study has demonstrated the potential of the MEMS sensor to determine sensitive viscosity changes within very small sample volumes, which could be of interest for various biotechnological applications.

Robust Chemiresistive Sensor for Continuous Monitoring of Free Chlorine Using Graphene-like Carbon.

Science.gov (United States)

Aryasomayajula, Aditya; Wojnas, Caroline; Divigalpitiya, Ranjith; Selvaganapathy, Ponnambalam Ravi; Kruse, Peter

2018-02-23

Free chlorine is widely used in industry as a bleaching and oxidizing agent. Its concentration is tightly monitored to avoid environmental contamination and deleterious human health effects. Here, we demonstrate a solid state chemiresistive sensor using graphene like carbon (GLC) to detect free chlorine in water. A 15-20 nm thick GLC layer on a PET substrate was modified with a redox-active aniline oligomer (phenyl-capped aniline tetramer, PCAT) to increase sensitivity, improve selectivity, and impart fouling resistance. Both the bare GLC sensor and the PCAT-modified GLC sensor can detect free chlorine continuously and, unlike previous chemiresistive sensors, do not require a reset. The PCAT-modified sensor showed a linear response with a slope of 13.89 (mg/L) -1 to free chlorine concentrations between 0.2 and 0.8 mg/L which is relevant for free chlorine monitoring for drinking water and wastewater applications. The PCAT-modified GLC sensors were found to be selective and showed less than 0.5% change in current in response to species such as nitrates, phosphates and sulfates in water. They also were resistant to fouling from organic material and showed only a 2% loss in signal. Tap water samples from residential area were tested using this sensor which showed good agreement with standard colorimetric measurement methods. The GLC and PCAT-GLC sensors show high sensitivity and excellent selectivity to free chlorine and can be used for continuous automated monitoring of free chlorine.

Development of a nonlinear model for the prediction of response times of glucose affinity sensors using concanavalin A and dextran and the development of a differential osmotic glucose affinity sensor

Science.gov (United States)

Reis, Louis G.

With the increasing prevalence of diabetes in the United States and worldwide, blood glucose monitoring must be accurate and reliable. Current enzymatic sensors have numerous disadvantages that make them unreliable and unfavorable among patients. Recent research in glucose affinity sensors correct some of the problems that enzymatic sensors experience. Dextran and concanavalin A are two of the more common components used in glucose affinity sensors. When these sensors were first explored, a model was derived to predict the response time of a glucose affinity sensor using concanavalin A and dextran. However, the model assumed the system was linear and fell short of calculating times representative of the response times determined through experimental tests with the sensors. In this work, a new model that uses the Stokes-Einstein Equation to demonstrate the nonlinear behavior of the glucose affinity assay was developed to predict the response times of similar glucose affinity sensors. In addition to the device tested by the original linear model, additional devices were identified and tested with the proposed model. The nonlinear model was designed to accommodate the many different variations between systems. The proposed model was able to accurately calculate response times for sensors using the concanavalin A-dextran affinity assay with respect to the experimentally reported times by the independent research groups. Parameter studies using the nonlinear model were able to identify possible setbacks that could compromise the response of thesystem. Specifically, the model showed that the improper use of asymmetrical membranes could increase the response time by as little as 20% or more as the device is miniaturized. The model also demonstrated that systems using the concanavalin Adextran assay would experience higher response times in the hypoglycemic range. This work attempted to replicate and improve an osmotic glucose affinity sensor. The system was designed to

Hydrogel-based electrochemical sensor for non-invasive and continuous glucose monitoring

Science.gov (United States)

Park, Habeen; Lee, Ji-Young; Kim, Dong-Chul; Koh, Younggook; Cha, Junhoe

2017-07-01

Monitoring blood glucose level of diabetic patients is crucial in diabetes care from life threating complications. Selfmonitoring blood glucose (SMBG) that involves finger prick to draw blood samples into the measurement system is a widely-used method of routine measurement of blood glucose levels to date. SMBG includes, however, unavoidable pain problems resulting from the repetitive measurements. We hereby present a hydrogel-based electrochemical (H-EC) sensor to monitor the glucose level, non-invasively. Glucose oxidase (GOx) was immobilized in the disc-type hydroxyethyl methacrylate (HEMA) based hydrogel and kept intact in the hydrogel. Fast electron transfer mediated by Prussian blue (PB, hexacyanoferrate) generated efficient signal amplifications to facilitate the detection of the extracted glucose from the interstitial fluid. The linear response and the selectivity against glucose of the H-EC sensor were validated by chronoamperometry. For the practical use, the outcomes from the correlation of the extracted glucose concentration and the blood glucose value by on-body extraction, as well as the validation of the hydrogel-based electrochemical (H-EC) device, were applied to the on-body glucose monitoring.

Modeling the relative impact of capsular tissue effects on implanted glucose sensor time lag and signal attenuation.

Science.gov (United States)

Novak, Matthew T; Yuan, Fan; Reichert, William M

2010-10-01

Little is known mechanistically about why implanted glucose sensors lag behind blood glucose levels in both the time to peak sensor response and the magnitude of peak sensor response. A mathematical model of glucose transport from capillaries through surrounding tissue to the sensor surface was constructed to address how different aspects of the tissue affect glucose transport to an implanted sensor. Physiologically relevant values of capsule diffusion coefficient, capsule porosity, cellular glucose consumption, capsule thickness, and subcutaneous vessel density were used as inputs to create simulated sensor traces that mimic experimental instances of time lag and concentration attenuation relative to a given blood glucose profile. Using logarithmic sensitivity analysis, each parameter was analyzed to study the effect of these variables on both lag and attenuation. Results identify capsule thickness as the strongest determinant of sensor time lag, while subcutaneous vessel density and capsule porosity had the largest effects on attenuation of glucose that reaches the sensor surface. These findings provide mechanistic insight for the rational design of sensor modifications that may alleviate the deleterious consequences of tissue effects on implanted sensor performance.

Highly sensitive glucose sensor based on monodisperse palladium nickel/activated carbon nanocomposites.

Science.gov (United States)

Koskun, Yağmur; Şavk, Aysun; Şen, Betül; Şen, Fatih

2018-06-20

Glucose enzyme biosensors have been used for a variety of applications such as medical diagnosis, bioprocess engineering, beverage industry and environmental scanning etc. and there is still a growing interest in glucose sensors. For this purpose, addressed herein, as a novel glucose sensor, highly sensitive activated carbon (AC) decorated monodisperse nickel and palladium alloy nanocomposites modified glassy carbon electrode (Ni-Pd@AC/GCE NCs) have been synthesized by in-situ reduction technique. Raman Spectroscopy (RS), X-ray Photoelectron Spectroscopy (XPS), X-ray Diffraction (XRD), Transmission Electron Microscopy (TEM), cyclic voltammetry (CV) and chronoamperometry (CA) were used for the characterization of the prepared non-enzymatic glucose sensor. The characteristic sensor properties of the Ni-Pd@AC/GCE electrode were compared with Ni-Pd NCs/GCE, Ni@AC/GCE and Pd@AC/GCE and the results demonstrate that the AC is very effective in the enhancement of the electrocatalytic properties of sensor. In addition, the Ni-Pd@AC/GCE nanocomposites showed a very low detection limit of 0.014 μM, a wide linear range of 0.01 mM-1 mM and a very high sensitivity of 90 mA mM -1  cm -2 . Furthermore, the recommended sensor offer the various advantageous such as facile preparation, fast response time, high selectivity and sensitivity. Lastly, monodisperse Ni-Pd@AC/GCE was utilized to detect glucose in real sample species. Copyright © 2018 Elsevier B.V. All rights reserved.

A contact lens with integrated telecommunication circuit and sensors for wireless and continuous tear glucose monitoring

International Nuclear Information System (INIS)

Yao, H; Liao, Y; Lingley, A R; Afanasiev, A; Lähdesmäki, I; Otis, B P; Parviz, B A

2012-01-01

We present an integrated functional contact lens, composed of a differential glucose sensor module, metal interconnects, sensor read-out circuit, antenna and telecommunication circuit, to monitor tear glucose levels wirelessly, continuously and non-invasively. The electrochemical differential sensor module is based on immobilization of activated and de-activated glucose oxidase. We characterized the sensor on a model polymer eye and determined that it showed good repeatability, molecular interference rejection and linearity in the range of 0–2 mM glucose, covering normal tear glucose concentrations (0.1–0.6 mM). We also report the temperature, ageing and protein-fouling sensitivity of the sensor. We report the design and implementation of a low-power (3 µW) sensor read-out and telecommunication circuit to deliver wireless power and transmit data for the sensor module. Using this small chip (0.36 mm 2 ), we produced an integrated contact lens with sensors and demonstrated wireless operation of the system and glucose read-out over the distance of several centimeters. (paper)

O-GlcNAcylation affects β-catenin and E-cadherin expression, cell motility and tumorigenicity of colorectal cancer.

Science.gov (United States)

Harosh-Davidovich, Shani Ben; Khalaila, Isam

2018-03-01

O-GlcNAcylation, the addition of β-N-acetylglucosamine (O-GlcNAc) moiety to Ser/Thr residues, is a sensor of the cell metabolic state. Cancer diseases such as colon, lung and breast cancer, possess deregulated O-GlcNAcylation. Studies during the last decade revealed that O-GlcNAcylation is implicated in cancer tumorigenesis and proliferation. The Wnt/β-catenin signaling pathway and cadherin-mediated adhesion are also implicated in epithelial-mesenchymal transition (EMT), a key cellular process in invasion and cancer metastasis. Often, deregulation of the Wnt pathway is caused by altered phosphorylation of its components. Specifically, phosphorylation of Ser or Thr residues of β-catenin affects its location and interaction with E-cadherin, thus facilitating cell-cell adhesion. Consistent with previous studies, the current study indicates that β-catenin is O-GlcNAcylated. To test the effect of O-GlcNAcylation on cell motility and how O-GlcNAcylation might affect β-catenin and E-cadherin functions, the enzyme machinery of O-GlcNAcylation was modulated either with chemical inhibitors or by gene silencing. When O-GlcNAcase (OGA) was inhibited, a global elevation of protein O-GlcNAcylation and increase in the expression of E-cadherin and β-catenin were noted. Concomitantly with enhanced O-GlcNAcylation, β-catenin transcriptional activity were elevated. Additionally, fibroblast cell motility was enhanced. Stable silenced cell lines with adenoviral OGA or adenoviral O-GlcNAc transferase (OGT) were established. Consistent with the results obtained by OGA chemical inhibition by TMG, OGT-silencing led to a significant reduction in β-catenin level. In vivo, murine orthotropic colorectal cancer model indicates that elevated O-GlcNAcylation leads to increased mortality rate, tumor and metastasis development. However, reduction in O-GlcNAcylation promoted survival that could be attributed to attenuated tumor and metastasis development. The results described herein provide

An Integrated Glucose Sensor with an All-Solid-State Sodium Ion-Selective Electrode for a Minimally Invasive Glucose Monitoring System

Directory of Open Access Journals (Sweden)

Junko Kojima

2015-06-01

Full Text Available We developed a minimally invasive glucose monitoring system that uses a microneedle to permeate the skin surface and a small hydrogel to accumulate interstitial fluid glucose. The measurement of glucose and sodium ion levels in the hydrogel is required for estimating glucose levels in blood; therefore, we developed a small, enzyme-fixed glucose sensor with a high-selectivity, all-solid-state, sodium ion-selective electrode (ISE integrated into its design. The glucose sensor immobilized glucose oxidase showed a good correlation between the glucose levels in the hydrogels and the reference glucose levels (r > 0.99, and exhibited a good precision (coefficient of variation = 2.9%, 0.6 mg/dL. In the design of the sodium ISEs, we used the insertion material Na0.33MnO2 as the inner contact layer and DD16C5 exhibiting high Na+/K+ selectivity as the ionophore. The developed sodium ISE exhibited high selectivity (\\( \\log \\,k^{pot}_{Na,K} = -2.8\\ and good potential stability. The sodium ISE could measure 0.4 mM (10−3.4 M sodium ion levels in the hydrogels containing 268 mM (10−0.57 M KCl. The small integrated sensor (ϕ < 10 mm detected glucose and sodium ions in hydrogels simultaneously within 1 min, and it exhibited sufficient performance for use as a minimally invasive glucose monitoring system.

Skin-Attachable, Stretchable Electrochemical Sweat Sensor for Glucose and pH Detection.

Science.gov (United States)

Oh, Seung Yun; Hong, Soo Yeong; Jeong, Yu Ra; Yun, Junyeong; Park, Heun; Jin, Sang Woo; Lee, Geumbee; Oh, Ju Hyun; Lee, Hanchan; Lee, Sang-Soo; Ha, Jeong Sook

2018-04-25

As part of increased efforts to develop wearable healthcare devices for monitoring and managing physiological and metabolic information, stretchable electrochemical sweat sensors have been investigated. In this study, we report on the fabrication of a stretchable and skin-attachable electrochemical sensor for detecting glucose and pH in sweat. A patterned stretchable electrode was fabricated via layer-by-layer deposition of carbon nanotubes (CNTs) on top of patterned Au nanosheets (AuNS) prepared by filtration onto stretchable substrate. For the detection of glucose and pH, CoWO 4 /CNT and polyaniline/CNT nanocomposites were coated onto the CNT-AuNS electrodes, respectively. A reference electrode was prepared via chlorination of silver nanowires. Encapsulation of the stretchable sensor with sticky silbione led to a skin-attachable sweat sensor. Our sensor showed high performance with sensitivities of 10.89 μA mM -1 cm -2 and 71.44 mV pH -1 for glucose and pH, respectively, with mechanical stability up to 30% stretching and air stability for 10 days. The sensor also showed good adhesion even to wet skin, allowing the detection of glucose and pH in sweat from running while being attached onto the skin. This work suggests the application of our stretchable and skin-attachable electrochemical sensor to health management as a high-performance healthcare wearable device.

Non-Enzymatic Wearable Sensor for Electrochemical Analysis of Perspiration Glucose.

Science.gov (United States)

Zhu, Xiaofei; Ju, Yinhui; Chen, Jian; Liu, Deye; Liu, Hong

2018-05-16

We report a non-enzymatic wearable sensor for electrochemical analysis of perspiration glucose. Multi-potential steps are applied on a Au electrode, including a high negative pretreatment potential step for proton reduction which produc-es a localized alkaline condition, a moderate potential step for electrocatalytic oxidation of glucose under the alkaline condi-tion, and a positive potential step to clean and reactivate the electrode surface for the next detection. Fluorocarbon-based materials were coated on the Au electrode for improving the selectivity and robustness of the sensor. A fully integrated wrist-band is developed for continuous real-time monitoring of perspiration glucose during physical activities, and uploading the test result to a Smartphone App via Bluetooth.

A Lab-on-a-Chip-Based Non-Invasive Optical Sensor for Measuring Glucose in Saliva

Directory of Open Access Journals (Sweden)

Dong Geon Jung

2017-11-01

Full Text Available A lab-on-a-chip (LOC-based non-invasive optical sensor for measuring glucose in saliva was fabricated. Existing glucose sensors utilizing blood require acquisition of a blood sample by pricking the finger, which is painful and inconvenient. To overcome these limitations, we propose a non-invasive glucose sensor with LOC, micro-electro-mechanical system and optical measurement technology. The proposed sensor for measuring glucose in saliva involves pretreatment, mixing, and measurement on a single tiny chip. Saliva containing glucose and glucose oxidase for glucose oxidation are injected through Inlets 1 and 2, respectively. Next, H2O2 is produced by the reaction between glucose and glucose oxidase in the pretreatment part. The saliva and generated H2O2 are mixed with a colorizing agent injected through Inlet 3 during the mixing part and the absorbance of the colorized mixture is measured in the measurement part. The absorbance of light increases as a function of glucose concentration at a wavelength of 630 nm. To measure the absorbance of the colorized saliva, a light-emitting diode with a wavelength of 630 nm and a photodiode were used during the measurement part. As a result, the measured output current of the photodiode decreased as glucose concentration in the saliva increased.

Dichloroacetate effects on glucose and lactate oxidation by neurons and astroglia in vitro and on glucose utilization by brain in vivo.

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Itoh, Yoshiaki; Esaki, Takanori; Shimoji, Kazuaki; Cook, Michelle; Law, Mona J; Kaufman, Elaine; Sokoloff, Louis

2003-04-15

Neuronal cultures in vitro readily oxidized both D-[(14)C]glucose and l-[(14)C]lactate to (14)CO(2), whereas astroglial cultures oxidized both substrates sparingly and metabolized glucose predominantly to lactate and released it into the medium. [(14)C]Glucose oxidation to (14)CO(2) varied inversely with unlabeled lactate concentration in the medium, particularly in neurons, and increased progressively with decreasing lactate concentration. Adding unlabeled glucose to the medium inhibited [(14)C]lactate oxidation to (14)CO(2) only in astroglia but not in neurons, indicating a kinetic preference in neurons for oxidation of extracellular lactate over intracellular pyruvatelactate produced by glycolysis. Protein kinase-catalyzed phosphorylation inactivates pyruvate dehydrogenase (PDH), which regulates pyruvate entry into the tricarboxylic acid cycle. Dichloroacetate inhibits this kinase, thus enhancing PDH activity. In vitro dichloroacetate stimulated glucose and lactate oxidation to CO(2) and reduced lactate release mainly in astroglia, indicating that limitations in glucose and lactate oxidation by astroglia may be due to a greater balance of PDH toward the inactive form. To assess the significance of astroglial export of lactate to neurons in vivo, we attempted to diminish this traffic in rats by administering dichloroacetate (50 mgkg) intravenously to stimulate astroglial lactate oxidation and then examined the effects on baseline and functionally activated local cerebral glucose utilization (lCMR(glc)). Dichloroacetate raised baseline lCMR(glc) throughout the brain and decreased the percent increases in lCMR(glc) evoked by functional activation. These studies provide evidence in support of the compartmentalization of glucose metabolism between astroglia and neurons but indicate that the compartmentalization may be neither complete nor entirely obligatory.

A microfluidic glucose sensor incorporating a novel thread-based electrode system.

Science.gov (United States)

Gaines, Michelle; Gonzalez-Guerrero, Maria Jose; Uchida, Kathryn; Gomez, Frank A

2018-05-01

An electrochemical sensor for the detection of glucose using thread-based electrodes and fabric is described. This device is relatively simple to fabricate and can be used for multiple readings after washing with ethanol. The fabrication of the chip consisted of two steps. First, three thread-based electrodes (reference, working, and counter) were fabricated by painting pieces of nylon thread with either layered silver ink and carbon ink or silver/silver chloride ink. The threads were then woven into a fabric chip with a beeswax barrier molded around the edges in order to prevent leaks from the tested solutions. A thread-based working electrode consisting of one layer of silver underneath two layers of carbon was selected to fabricate the final sensor system. Using the chip, a PBS solution containing glucose oxidase (GOx) (10 mg/mL), potassium ferricyanide (K 3 [Fe(CN) 6 ]) (10 mg/mL) as mediator, and different concentrations of glucose (0-25 mM), was measured by cyclic voltammetry (CV). It was found that the current output from the oxidation of glucose was proportional to the glucose concentrations. This thread-based electrode system is a viable sensor platform for detecting glucose in the physiological range. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Supraoptic oxytocin and vasopressin neurons function as glucose and metabolic sensors.

Science.gov (United States)

Song, Zhilin; Levin, Barry E; Stevens, Wanida; Sladek, Celia D

2014-04-01

Neurons in the supraoptic nuclei (SON) produce oxytocin and vasopressin and express insulin receptors (InsR) and glucokinase. Since oxytocin is an anorexigenic agent and glucokinase and InsR are hallmarks of cells that function as glucose and/or metabolic sensors, we evaluated the effect of glucose, insulin, and their downstream effector ATP-sensitive potassium (KATP) channels on calcium signaling in SON neurons and on oxytocin and vasopressin release from explants of the rat hypothalamo-neurohypophyseal system. We also evaluated the effect of blocking glucokinase and phosphatidylinositol 3 kinase (PI3K; mediates insulin-induced mobilization of glucose transporter, GLUT4) on responses to glucose and insulin. Glucose and insulin increased intracellular calcium ([Ca(2+)]i). The responses were glucokinase and PI3K dependent, respectively. Insulin and glucose alone increased vasopressin release (P glucose in the presence of insulin. The oxytocin (OT) and vasopressin (VP) responses to insulin+glucose were blocked by the glucokinase inhibitor alloxan (4 mM; P ≤ 0.002) and the PI3K inhibitor wortmannin (50 nM; OT: P = 0.03; VP: P ≤ 0.002). Inactivating K ATP channels with 200 nM glibenclamide increased oxytocin and vasopressin release (OT: P neurons functioning as glucose and "metabolic" sensors to participate in appetite regulation.

Supraoptic oxytocin and vasopressin neurons function as glucose and metabolic sensors

Science.gov (United States)

Song, Zhilin; Levin, Barry E.; Stevens, Wanida

2014-01-01

Neurons in the supraoptic nuclei (SON) produce oxytocin and vasopressin and express insulin receptors (InsR) and glucokinase. Since oxytocin is an anorexigenic agent and glucokinase and InsR are hallmarks of cells that function as glucose and/or metabolic sensors, we evaluated the effect of glucose, insulin, and their downstream effector ATP-sensitive potassium (KATP) channels on calcium signaling in SON neurons and on oxytocin and vasopressin release from explants of the rat hypothalamo-neurohypophyseal system. We also evaluated the effect of blocking glucokinase and phosphatidylinositol 3 kinase (PI3K; mediates insulin-induced mobilization of glucose transporter, GLUT4) on responses to glucose and insulin. Glucose and insulin increased intracellular calcium ([Ca2+]i). The responses were glucokinase and PI3K dependent, respectively. Insulin and glucose alone increased vasopressin release (P glucose in the presence of insulin. The oxytocin (OT) and vasopressin (VP) responses to insulin+glucose were blocked by the glucokinase inhibitor alloxan (4 mM; P ≤ 0.002) and the PI3K inhibitor wortmannin (50 nM; OT: P = 0.03; VP: P ≤ 0.002). Inactivating KATP channels with 200 nM glibenclamide increased oxytocin and vasopressin release (OT: P neurons functioning as glucose and “metabolic” sensors to participate in appetite regulation. PMID:24477542

O-GlcNAcylation of cardiac Nav1.5 contributes to the development of arrhythmias in diabetic hearts.

Science.gov (United States)

Yu, Peng; Hu, Lili; Xie, Jinyan; Chen, Sisi; Huang, Lin; Xu, Zixuan; Liu, Xiao; Zhou, Qiongqiong; Yuan, Ping; Yan, Xia; Jin, Jiejin; Shen, Yang; Zhu, Wengen; Fu, Linghua; Chen, Qi; Yu, Jianhua; Hu, Jianxin; Cao, Qing; Wan, Rong; Hong, Kui

2018-06-01

Cardiovascular complications are major causes of mortality and morbidity in diabetic patients. The mechanisms underlying the progression of diabetic heart (DH) to ventricular arrhythmias are unclear. O-linked GlcNAcylation (O-GlcNAc) is a reversible post-translational modification for the regulation of diverse cellular processes. The purpose of this study was to assess whether the cardiac voltage-gated sodium channel (Nav1.5) is subjected to O-linked GlcNAcylation (O-GlcNAc), which plays an essential role in DH-induced arrhythmias. In this study, Sprague-Dawley rats (male, 200-230 g) were treated with a single high-dose of streptozotocin (STZ, 80 mg/kg) to generate a rat model of diabetes. STZ-induced 3-month diabetic rats displayed increased susceptibility to ventricular arrhythmias. The elevated O-GlcNAc modification was correlated with decreases in both total and cytoplasmic Nav1.5 expression in vivo and in vitro. In addition, both co-immunoprecipitation and immunostaining assays demonstrated that hyperglycemia could increase the O-GlcNAc-modified Nav1.5 levels and decrease the interaction between Nav1.5 and Nav1.5-binding proteins Nedd4-2/SAP-97. Furthermore, patch-clamp measurements in HEK-293 T cells showed that Nav1.5 current densities decreased by 30% after high-glucose treatment, and the sodium currents increased via O-GlcNAc inhibition. Our data suggested that hyperglycemia increased the O-GlcNAc modification of Nav1.5 expression and decreased the interaction between Nav1.5 and Nedd4-2/SAP-97, which led to the abnormal expression and distribution of Nav1.5, loss of function of the sodium channel, and prolongation of the PR/QT interval. Excessive O-GlcNAc modification of Nav1.5 is a novel signaling event, which may be an underlying contributing factor for the development of the arrhythmogenesis in DH. Copyright © 2017. Published by Elsevier B.V.

Enzymatic and non-enzymatic electrochemical glucose sensor based on carbon nano-onions

Science.gov (United States)

Mohapatra, Jeotikanta; Ananthoju, Balakrishna; Nair, Vishnu; Mitra, Arijit; Bahadur, D.; Medhekar, N. V.; Aslam, M.

2018-06-01

A high sensitive glucose sensing characteristic has been realized in carbon nano-onions (CNOs). The CNOs of mean size 30 nm were synthesized by an energy-efficient, simple and inexpensive combustion technique. These as-synthesized CNOs could be employed as an electrochemical sensor by covalently immobilizing the glucose oxidase enzyme on them via carbodiimide chemistry. The sensitivity achieved by such a sensor is 26.5 μA mM-1 cm-2 with a linear response in the range of 1-10 mM glucose. Further to improve the catalytic activity of the CNOs and also to make them enzyme free, platinum nanoparticles of average size 2.5 nm are decorated on CNOs. This sensor fabricated using Pt-decorated CNOs (Pt@CNOs) nanostructure has shown an enhanced sensitivity of 21.6 μA mM-1 cm-2 with an extended linear response in the range of 2-28 mM glucose. Through these attempts we demonstrate CNOs as a versatile biosensing platform.

Novel Dry-Type Glucose Sensor Based on a Metal-Oxide-Semiconductor Capacitor Structure with Horseradish Peroxidase + Glucose Oxidase Catalyzing Layer

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Lin, Jing-Jenn; Wu, You-Lin; Hsu, Po-Yen

2007-10-01

In this paper, we present a novel dry-type glucose sensor based on a metal-oxide-semiconductor capacitor (MOSC) structure using SiO2 as a gate dielectric in conjunction with a horseradish peroxidase (HRP) + glucose oxidase (GOD) catalyzing layer. The tested glucose solution was dropped directly onto the window opened on the SiO2 layer, with a coating of HRP + GOD catalyzing layer on top of the gate dielectric. From the capacitance-voltage (C-V) characteristics of the sensor, we found that the glucose solution can induce an inversion layer on the silicon surface causing a gate leakage current flowing along the SiO2 surface. The gate current changes Δ I before and after the drop of glucose solution exhibits a near-linear relationship with increasing glucose concentration. The Δ I sensitivity is about 1.76 nA cm-2 M-1, and the current is quite stable 20 min after the drop of the glucose solution is tested.

Enhancing the accuracy of subcutaneous glucose sensors: a real-time deconvolution-based approach.

Science.gov (United States)

Guerra, Stefania; Facchinetti, Andrea; Sparacino, Giovanni; Nicolao, Giuseppe De; Cobelli, Claudio

2012-06-01

Minimally invasive continuous glucose monitoring (CGM) sensors can greatly help diabetes management. Most of these sensors consist of a needle electrode, placed in the subcutaneous tissue, which measures an electrical current exploiting the glucose-oxidase principle. This current is then transformed to glucose levels after calibrating the sensor on the basis of one, or more, self-monitoring blood glucose (SMBG) samples. In this study, we design and test a real-time signal-enhancement module that, cascaded to the CGM device, improves the quality of its output by a proper postprocessing of the CGM signal. In fact, CGM sensors measure glucose in the interstitium rather than in the blood compartment. We show that this distortion can be compensated by means of a regularized deconvolution procedure relying on a linear regression model that can be updated whenever a pair of suitably sampled SMBG references is collected. Tests performed both on simulated and real data demonstrate a significant accuracy improvement of the CGM signal. Simulation studies also demonstrate the robustness of the method against departures from nominal conditions, such as temporal misplacement of the SMBG samples and uncertainty in the blood-to-interstitium glucose kinetic model. Thanks to its online capabilities, the proposed signal-enhancement algorithm can be used to improve the performance of CGM-based real-time systems such as the hypo/hyper glycemic alert generators or the artificial pancreas.

A Chemoenzymatic Histology Method for O-GlcNAc Detection.

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Aguilar, Aime Lopez; Hou, Xiaomeng; Wen, Liuqing; Wang, Peng G; Wu, Peng

2017-12-14

Modification of nuclear and cytoplasmic proteins by the addition or removal of O-GlcNAc dynamically impacts multiple biological processes. Here, we present the development of a chemoenzymatic histology method for the detection of O-GlcNAc in tissue specimens. We applied this method to screen murine organs, uncovering specific O-GlcNAc distribution patterns in different tissue structures. We then utilized our histology method for O-GlcNAc detection in human brain specimens from healthy donors and donors with Alzheimer's disease and found higher levels of O-GlcNAc in specimens from healthy donors. We also performed an analysis using a multiple cancer tissue array, uncovering different O-GlcNAc levels between healthy and cancerous tissues, as well as different O-GlcNAc cellular distributions within certain tissue specimens. This chemoenzymatic histology method therefore holds great potential for revealing the biology of O-GlcNAc in physiopathological processes. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Design strategy of a graphene based bio-sensor for glucose

DEFF Research Database (Denmark)

Cantatore, Valentina; Pandit, Santosh; Mokkapati, V.R.S.S.

2018-01-01

A novel graphene-based glucose sensor-design is formulated and explored in silico. An ad hoc host molecule is tailored to bind to glucose by multiple hydrogen bonds. A pyridinic core is chosen for this receptor in order to allow for “socket-plug” dative bonding to boron sites of boron doped...

O-GlcNAc profiling: from proteins to proteomes

Science.gov (United States)

2014-01-01

O-linked β-D-N-acetylglucosamine (O-GlcNAc) modification (O-GlcNAcylation) onto serine and threonine residues of proteins is an important post-translational modification (PTM), which is involved in many crucial biological processes including transcription, translation, proteasomal degradation, and signal transduction. Aberrant protein O-GlcNAcylation is directly linked to the pathological progression of chronic diseases including diabetes, cancer, and neurodegenerative disorders. Identification, site mapping, and quantification of O-GlcNAc proteins are a prerequisite to decipher their functions. In this review, we mainly focus on technological developments regarding O-GlcNAc protein profiling. Specifically, on one hand, we show how these techniques are being used for the comprehensive characterization of certain targeted proteins in which biologists are most interested. On the other hand, we present several newly developed approaches for O-GlcNAcomic profiling as well as how they provide us with a systems perspective to crosstalk amongst different PTMs and complicated biological events. Promising technical trends are also highlighted to evoke more efforts by diverse laboratories, which would further expand our understanding of the physiological and pathological roles of protein O-GlcNAcylation in chronic diseases. PMID:24593906

Glucose supplementation does not interfere with fasting-induced protection against renal ischemia/reperfusion injury in mice.

Science.gov (United States)

Verweij, Mariëlle; van de Ven, Marieke; Mitchell, James R; van den Engel, Sandra; Hoeijmakers, Jan H J; Ijzermans, Jan N M; de Bruin, Ron W F

2011-10-15

Preoperative fasting induces robust protection against renal ischemia/reperfusion (I/R) injury in mice but is considered overcautious and possibly detrimental for postoperative recovery in humans. Furthermore, fasting seems to conflict with reported benefits of preoperative nutritional enhancement with carbohydrate-rich drinks. Here, we investigated whether preoperative ingestion of a glucose solution interferes with fasting-induced protection against renal I/R injury. Mice were randomized into the following groups: fasted for 3 days with access to water (fasted) or a glucose solution (fasted+glc) and fed ad libitum with water (fed) or a glucose solution (fed+glc). After induction of bilateral renal I/R injury, all animals had free access to food and water. Calorie intake, body weight, insulin sensitivity, kidney function, and animal survival were determined. Fed+glc mice had a comparable daily calorie intake as fed mice, but 50% of those calories were obtained from the glucose solution. Fasted+glc mice had a daily calorie intake of approximately 75% of the intake of both fed groups. This largely prevented the substantial body weight loss seen in fasted animals. Preoperative insulin sensitivity was significantly improved in fasted+glc mice versus fed mice. After I/R injury, kidney function and animal survival were superior in both fasted groups. The benefits of fasting and preoperative nutritional enhancement with carbohydrates are not mutually exclusive and may be a clinically feasible regimen to protect against renal I/R injury.

MWCNT-ruthenium oxide composite paste electrode as non-enzymatic glucose sensor.

Science.gov (United States)

Tehrani, Ramin M A; Ab Ghani, Sulaiman

2012-01-01

A non-enzymatic glucose sensor of multi-walled carbon nanotube-ruthenium oxide/composite paste electrode (MWCNT-RuO(2)/CPE) was developed. The electrode was characterized by using XRD, SEM, TEM and EIS. Meanwhile, cyclic voltammetry and amperometry were used to check on the performances of the MWCNT-RuO(2)/CPE towards glucose. The proposed electrode has displayed a synergistic effect of RuO(2) and MWCNT on the electrocatalytic oxidation of glucose in 3M NaOH. This was possible via the formation of transitions of two redox pairs, viz. Ru(VI)/Ru(IV) and Ru(VII)/Ru(VI). A linear range of 0.5-50mM glucose and a limit of detection of 33 μM glucose (S/N=3) were observed. There was no significant interference observable from the traditional interferences, viz. ascorbic acid and uric acid. Indeed, results so obtained have indicated that the developed MWCNT-RuO(2)/CPE would pave the way for a better future to glucose sensor development as its fabrication was without the use of any enzyme. Copyright © 2012 Elsevier B.V. All rights reserved.

Continuous glucose monitoring--a study of the Enlite sensor during hypo- and hyperbaric conditions.

Science.gov (United States)

Adolfsson, Peter; Örnhagen, Hans; Eriksson, Bengt M; Cooper, Ken; Jendle, Johan

2012-06-01

The performance and accuracy of the Enlite(™) (Medtronic, Inc., Northridge, CA) sensor may be affected by microbubble formation at the electrode surface during hypo- and hyperbaric conditions. The effects of acute pressure changes and of prewetting of sensors were investigated. On Day 1, 24 sensors were inserted on the right side of the abdomen and back in one healthy individual; 12 were prewetted with saline solution, and 12 were inserted dry. On Day 2, this procedure was repeated on the left side. All sensors were attached to an iPro continuous glucose monitoring (CGM) recorder. Hypobaric and hyperbaric tests were conducted in a pressure chamber, with each test lasting 105 min. Plasma glucose values were obtained at 5-min intervals with a HemoCue(®) (Ängelholm, Sweden) model 201 glucose analyzer for comparison with sensor glucose values. Ninety percent of the CGM systems operated during the tests. The mean absolute relative difference was lower during hyperbaric than hypobaric conditions (6.7% vs. 14.9%, Phypobaric but not during hyperbaric conditions. Clarke Error Grid Analysis showed that 100% of the values were found in the A+B region. No differences were found between prewetted and dry sensors. The Enlite sensor performed adequately during acute pressure changes and was more accurate during hyperbaric than hypobaric conditions. Prewetting the sensors did not improve accuracy. Further studies on type 1 diabetes subjects are needed under various pressure conditions.

Monoaminergic Control of Cellular Glucose Utilization by Glycogenolysis in Neocortex and Hippocampus.

Science.gov (United States)

DiNuzzo, Mauro; Giove, Federico; Maraviglia, Bruno; Mangia, Silvia

2015-12-01

Brainstem nuclei are the principal sites of monoamine (MA) innervation to major forebrain structures. In the cortical grey matter, increased secretion of MA neuromodulators occurs in response to a wealth of environmental and homeostatic challenges, whose onset is associated with rapid, preparatory changes in neural activity as well as with increases in energy metabolism. Blood-borne glucose is the main substrate for energy production in the brain. Once entered the tissue, interstitial glucose is equally accessible to neurons and astrocytes, the two cell types accounting for most of cellular volume and energy metabolism in neocortex and hippocampus. Astrocytes also store substantial amounts of glycogen, but non-stimulated glycogen turnover is very small. The rate of cellular glucose utilization in the brain is largely determined by hexokinase, which under basal conditions is more than 90 % inhibited by its product glucose-6-phosphate (Glc-6-P). During rapid increases in energy demand, glycogen is a primary candidate in modulating the intracellular level of Glc-6-P, which can occur only in astrocytes. Glycogenolysis can produce Glc-6-P at a rate higher than uptake and phosphorylation of glucose. MA neurotransmitter are released extrasinaptically by brainstem neurons projecting to neocortex and hippocampus, thus activating MA receptors located on both neuronal and astrocytic plasma membrane. Importantly, MAs are glycogenolytic agents and thus they are exquisitely suitable for regulation of astrocytic Glc-6-P concentration, upstream substrate flow through hexokinase and hence cellular glucose uptake. Conforming to such mechanism, Gerald A. Dienel and Nancy F. Cruz recently suggested that activation of noradrenergic locus coeruleus might reversibly block astrocytic glucose uptake by stimulating glycogenolysis in these cells, thereby anticipating the rise in glucose need by active neurons. In this paper, we further develop the idea that the whole monoaminergic system

Chemical Arsenal for the Study of O-GlcNAc

Directory of Open Access Journals (Sweden)

Eun J. Kim

2011-02-01

Full Text Available The concepts of both protein glycosylation and cellular signaling have been influenced by O-linked-β-N-acetylglucosamine (O-GlcNAc modification (O-GlcNAcylation on the hydroxyl group of serine or threonine residues. Unlike conventional protein glycosylation, O-GlcNAcylation is localized in the nucleocytoplasm and its cycling is a dynamic process that operates in a highly regulated manner in response to various cellular stimuli. These characteristics render O-GlcNAcylation similar to phosphorylation, which has long been considered a major regulatory mechanism in cellular processes. Various efficient chemical approaches and novel mass spectrometric (MS techniques have uncovered numerous O-GlcNAcylated proteins that are involved in the regulation of many important cellular events. These discoveries imply that O-GlcNAcylation is another major regulator of cellular signaling. However, in contrast to phosphorylation, which is regulated by hundreds of kinases and phosphatases, dynamic O-GlcNAc cycling is catalyzed by only two enzymes: uridine diphospho-N-acetyl-glucosamine:polypeptide β-N-acetylglucosaminyl transferase (OGT and β-D-N-acetylglucosaminidase (OGA. Many useful chemical tools have recently been used to greatly expand our understanding of the extensive crosstalk between O-GlcNAcylation and phosphorylation and hence of cellular signaling. This review article describes the various useful chemical tools that have been developed and discusses the considerable advances made in the O-GlcNAc field.

Identification of O-GlcNAcylated proteins in Plasmodium falciparum.

Science.gov (United States)

Kupferschmid, Mattis; Aquino-Gil, Moyira Osny; Shams-Eldin, Hosam; Schmidt, Jörg; Yamakawa, Nao; Krzewinski, Frédéric; Schwarz, Ralph T; Lefebvre, Tony

2017-11-29

Post-translational modifications (PTMs) constitute a huge group of chemical modifications increasing the complexity of the proteomes of living beings. PTMs have been discussed as potential anti-malarial drug targets due to their involvement in many cell processes. O-GlcNAcylation is a widespread PTM found in different organisms including Plasmodium falciparum. The aim of this study was to identify O-GlcNAcylated proteins of P. falciparum, to learn more about the modification process and to understand its eventual functions in the Apicomplexans. The P. falciparum strain 3D7 was amplified in erythrocytes and purified. The proteome was checked for O-GlcNAcylation using different methods. The level of UDP-GlcNAc, the donor of the sugar moiety for O-GlcNAcylation processes, was measured using high-pH anion exchange chromatography. O-GlcNAcylated proteins were enriched and purified utilizing either click chemistry labelling or adsorption on succinyl-wheat germ agglutinin beads. Proteins were then identified by mass-spectrometry (nano-LC MS/MS). While low when compared to MRC5 control cells, P. falciparum disposes of its own pool of UDP-GlcNAc. By using proteomics methods, 13 O-GlcNAcylated proteins were unambiguously identified (11 by click-chemistry and 6 by sWGA-beads enrichment; 4 being identified by the 2 approaches) in late trophozoites. These proteins are all part of pathways, functions and structures important for the parasite survival. By probing clicked-proteins with specific antibodies, Hsp70 and α-tubulin were identified as P. falciparum O-GlcNAc-bearing proteins. This study is the first report on the identity of P. falciparum O-GlcNAcylated proteins. While the parasite O-GlcNAcome seems close to those of other species, the structural differences exhibited by the proteomes provides a glimpse of innovative therapeutic paths to fight malaria. Blocking biosynthesis of UDP-GlcNAc in the parasites is another promising option to reduce Plasmodium life cycle.

Nonenzymatic glucose sensor based on disposable pencil graphite electrode modified by copper nanoparticles

Directory of Open Access Journals (Sweden)

Sima Pourbeyram

2016-10-01

Full Text Available A nonenzymatic glucose sensor based on a disposable pencil graphite electrode (PGE modified by copper nanoparticles [Cu(NP] was prepared for the first time. The prepared Cu(NP exhibited an absorption peak centered at ∼562 nm using UV-visible spectrophotometry and an almost homogenous spherical shape by scanning electron microscopy. Cyclic voltammetry of Cu(NP-PGE showed an adsorption controlled charge transfer process up to 90.0 mVs−1. The sensor was applied for the determination of glucose using an amperometry technique with a detection limit of [0.44 (±0.01 μM] and concentration sensitivity of [1467.5 (±1.3 μA/mMcm−2]. The preparation of the Cu(NP-PGE sensor was reproducible (relative standard deviation = 2.10%, n = 10, very simple, fast, and inexpensive, and the Cu(NP-PGE is suitable to be used as a disposable glucose sensor.

O-GlcNAcylation of RACK1 promotes hepatocellular carcinogenesis.

Science.gov (United States)

Duan, Fangfang; Wu, Hao; Jia, Dongwei; Wu, Weicheng; Ren, Shifang; Wang, Lan; Song, Shushu; Guo, Xinying; Liu, Fenglin; Ruan, Yuanyuan; Gu, Jianxin

2018-06-01

Aberrant oncogenic mRNA translation and protein O-linked β-N-acetylglucosaminylation (O-GlcNAcylation) are general features during tumorigenesis. Nevertheless, whether and how these two pathways are interlinked remain unknown. Our previous study indicated that ribosomal receptor for activated C-kinase 1 (RACK1) promoted chemoresistance and growth in hepatocellular carcinoma (HCC). The aim of this study is to examine the role of RACK1 O-GlcNAcylation in oncogene translation and HCC carcinogenesis. The site(s) of RACK1 for O-GlcNAcylation was mapped by mass spectrometry analysis. HCC cell lines were employed to examine the effects of RACK1 O-GlcNAcylation on the translation of oncogenic factors and behaviors of tumor cells in vitro. Transgenic knock-in mice were used to detect the role of RACK1 O-GlcNAcylation in modulating HCC tumorigenesis in vivo. The correlation of RACK1 O-GlcNAcylation with tumor progression and relapse were analyzed in clinical HCC samples. We found that ribosomal RACK1 was highly modified by O-GlcNAc at Ser122. O-GlcNAcylation of RACK1 enhanced its protein stability, ribosome binding and interaction with PKCβII (PRKCB), leading to increased eukaryotic translation initiation factor 4E phosphorylation and translation of potent oncogenes in HCC cells. Genetic ablation of RACK1 O-GlcNAcylation at Ser122 dramatically suppressed tumorigenesis, angiogenesis, and metastasis in vitro and in diethylnitrosamine (DEN)-induced HCC mouse model. Increased RACK1 O-GlcNAcylation was also observed in HCC patient samples and correlated with tumor development and recurrence after chemotherapy. These findings demonstrate that RACK1 acts as key mediator linking O-GlcNAc metabolism to cap-dependent translation during HCC tumorigenesis. Targeting RACK1 O-GlcNAcylation provides promising options for HCC treatment. O-GlcNAcylation of ribosomal receptor for activated C-kinase 1 at the amino acid serine122 promotes its stability, ribosome localization and interaction

O-GlcNAcylation in oral squamous cell carcinoma.

Science.gov (United States)

Kongkaew, Tassaporn; Aung, Win Pa Pa; Supanchart, Chayarop; Makeudom, Anupong; Langsa-Ard, Sarawat; Sastraruji, Thanapat; Chaiyarit, Ponlatham; Krisanaprakornkit, Suttichai

2018-03-01

Two post-translational mechanisms commonly demonstrated in various cancers are protein phosphorylation and glycosylation by O-linked β-N-acetylglucosamine (O-GlcNAc). However, only phosphorylation of the epidermal growth factor receptor (EGFR)/Akt pathway has been reported in oral squamous cell carcinoma (OSCC). Therefore, we aimed to determine both post-translational modifications in OSCC tissues and in oral cancer cells compared to normal tissues and oral keratinocytes and to find correlations of these modifications with histological grading. Thirty-two OSCC and ten normal formalin-fixed and paraffin-embedded sections were probed with the anti-O-GlcNAc, anti-O-GlcNAc transferase (OGT), anti-phosphorylated-EGFR tyr1173 , and anti-phosphorylated-Akt ser473 antibodies following standard immunohistochemistry. The immunohistochemical (IHC) score was determined using the Fromowitz standard. Whole cell lysates of oral cancer cells and normal oral keratinocytes were immunoblotted with the anti-O-GlcNAc antibody. The median IHC scores of O-GlcNAc or OGT between OSCC and normal tissues were not different, whereas those of phosphorylated-EGFR tyr1173 and phosphorylated-Akt ser473 were significantly higher in OSCC than normal tissues (P O-GlcNAcylated proteins in oral cancer cells and normal oral keratinocytes did not differ. In the OSCC group, the median IHC scores of O-GlcNAc and OGT were significantly lower than those of phosphorylated-EGFR tyr1173 and phosphorylated-Akt ser473 (P O-GlcNAc or OGT were not determined to correlate with histological grading. Unlike other types of cancers, our findings demonstrate that the levels of O-GlcNAcylation are not significantly increased in OSCC tissues or in oral cancer cells and are not associated with the histological grading of OSCC. © 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Synthesis, optimization and structural characterization of a chitosan-glucose derivative obtained by the Maillard reaction.

Science.gov (United States)

Gullón, Beatriz; Montenegro, María I; Ruiz-Matute, Ana I; Cardelle-Cobas, Alejandra; Corzo, Nieves; Pintado, Manuela E

2016-02-10

Chitosan (Chit) was submitted to the Maillard reaction (MR) by co-heating a solution with glucose (Glc). Different reaction conditions as temperature (40, 60 and 80 °C), Glc concentration (0.5%, 1%, and 2%, w/v), and reaction time (72, 52 and 24h) were evaluated. Assessment of the reaction extent was monitored by measuring changes in UV absorbance, browning and fluorescence. Under the best conditions, 2% (w/v) of Chit, 2% (w/v) of Glc at 60°C and 32 h of reaction time, a chitosan-glucose (Chit-Glc) derivative was purified and submitted to structural characterization to confirm its formation. Analysis of its molecular weight (MW) and the degree of substitution (DS) was carried out by HPLC-Size Exclusion Chromatography (SEC) and a colloid titration method, respectively. FT-IR and (1)H NMR were also used to analyze the functional groups and evaluate the introduction of Glc into the Chit molecule. According to our objectives, the results obtained in this work allowed to better understand the key parameters influencing the MR with Chit as well as to confirm the successful introduction of Glc into the Chit molecule obtaining a Chit-Glc derivative with a DS of 64.76 ± 4.40% and a MW of 210.37 kDa. Copyright © 2015 Elsevier Ltd. All rights reserved.

Analysis of Protein O-GlcNAcylation by Mass Spectrometry

OpenAIRE

Ma, Junfeng; Hart, Gerald W.

2017-01-01

O-linked β-D-N-acetylglucosamine (O-GlcNAc) addition (O-GlcNAcylation), a post-translational modification of serine/threonine residues of proteins, is involved in diverse cellular metabolic and signaling pathways. Aberrant O-GlcNAcylation underlies the initiation and progression of multiple chronic diseases including diabetes, cancer, and neurodegenerative diseases. Numerous methods have been developed for the analysis of protein O-GlcNAcylation, but instead of discussing the classical bioche...

A flexible and highly sensitive nonenzymatic glucose sensor based on DVD-laser scribed graphene substrate.

Science.gov (United States)

Lin, Songyue; Feng, Wendou; Miao, Xiaofei; Zhang, Xiangxin; Chen, Sujing; Chen, Yuanqiang; Wang, Wei; Zhang, Yining

2018-07-01

Flexible and implantable glucose biosensors are emerging technologies for continuous monitoring of blood-glucose of diabetes. Developing a flexible conductive substrates with high active surface area is critical for advancing the technology. Here, we successfully fabricate a flexible and highly sensitive nonenzymatic glucose by using DVD-laser scribed graphene (LSG) as a flexible conductively substrate. Copper nanoparticles (Cu-NPs) are electrodeposited as the catalyst. The LSG/Cu-NPs sensor demonstrates excellent catalytic activity toward glucose oxidation and exhibits a linear glucose detection range from 1 μM to 4.54 mM with high sensitivity (1.518 mA mM -1 cm -2 ) and low limit of detection (0.35 μM). Moreover, the LSG/Cu-NPs sensor shows excellent reproducibility and long-term stability. It is also highly selective toward glucose oxidation under the presence of various interfering species. Excellent flexing stability is also demonstrated by the LSG/Cu-NPs sensor, which is capable of maintaining 83.9% of its initial current after being bent against a 4-mm diameter rod for 180 times. The LSG/Cu-NPs sensor shows great potential for practical application as a nonenzymatic glucose biosensor. Meanwhile, the LSG conductive substrate provides a platform for the developing next-generation flexible and potentially implantable bioelectronics and biosensors. Copyright © 2018 Elsevier B.V. All rights reserved.

A novel strategy for global mapping of O-GlcNAc proteins and peptides using selective enzymatic deglycosylation, HILIC enrichment and mass spectrometry identification.

Science.gov (United States)

Shen, Bingquan; Zhang, Wanjun; Shi, Zhaomei; Tian, Fang; Deng, Yulin; Sun, Changqing; Wang, Guangshun; Qin, Weijie; Qian, Xiaohong

2017-07-01

O-GlcNAcylation is a kind of dynamic O-linked glycosylation of nucleocytoplasmic and mitochondrial proteins. It serves as a major nutrient sensor to regulate numerous biological processes including transcriptional regulation, cell metabolism, cellular signaling, and protein degradation. Dysregulation of cellular O-GlcNAcylated levels contributes to the etiologies of many diseases such as diabetes, neurodegenerative disease and cancer. However, deeper insight into the biological mechanism of O-GlcNAcylation is hampered by its extremely low stoichiometry and the lack of efficient enrichment approaches for large-scale identification by mass spectrometry. Herein, we developed a novel strategy for the global identification of O-GlcNAc proteins and peptides using selective enzymatic deglycosylation, HILIC enrichment and mass spectrometry analysis. Standard O-GlcNAc peptides can be efficiently enriched even in the presence of 500-fold more abundant non-O-GlcNAc peptides and identified by mass spectrometry with a low nanogram detection sensitivity. This strategy successfully achieved the first large-scale enrichment and characterization of O-GlcNAc proteins and peptides in human urine. A total of 474 O-GlcNAc peptides corresponding to 457 O-GlcNAc proteins were identified by mass spectrometry analysis, which is at least three times more than that obtained by commonly used enrichment methods. A large number of unreported O-GlcNAc proteins related to cell cycle, biological regulation, metabolic and developmental process were found in our data. The above results demonstrated that this novel strategy is highly efficient in the global enrichment and identification of O-GlcNAc peptides. These data provide new insights into the biological function of O-GlcNAcylation in human urine, which is correlated with the physiological states and pathological changes of human body and therefore indicate the potential of this strategy for biomarker discovery from human urine. Copyright �

In-vitro performance of the Enlite Sensor in various glucose concentrations during hypobaric and hyperbaric conditions.

Science.gov (United States)

Adolfsson, Peter; Ornhagen, Hans; Eriksson, Bengt M; Gautham, Raghavendhar; Jendle, Johan

2012-11-01

There is a need for reliable methods of glucose measurement in different environmental conditions. The objective of this in vitro study was to evaluate the performance of the Enlite® Sensor when connected to either the iPro™ Continuous Glucose Monitor recording device or the Guardian® REAL-Time transmitting device, in hypobaric and hyperbaric conditions. Sixteen sensors connected to eight iPro devices and eight Guardian REAL-Time devices were immersed in three beakers containing separate glucose concentrations: 52, 88, and 207 mg/dl (2.9, 4.9, and 11.3 mmol/liter). Two different pressure tests were conducted: a hypobaric test, corresponding to maximum 18000 ft/5500 m height, and a hyperbaric test, corresponding to maximum 100 ft/30 m depth. The linearity of the sensor signals in the different conditions was evaluated. The sensors worked continuously, and the sensor signals were collected without interruption at all pressures tested. When comparing the input signals for glucose (ISIGs) and the different glucose concentrations during altered pressure, linearity (R(2)) of 0.98 was found. During the hypobaric test, significant differences (p hyperbaric test, no differences were found. The Enlite Sensors connected to either the iPro or the Guardian REAL-Time device provided values continuously. In hyperbaric conditions, no significant differences were seen during changes in ambient pressure; however, during hypobaric conditions, the ISIG was significantly different in the low and high glucose concentrations. © 2012 Diabetes Technology Society.

An extremely sensitive monoboronic acid based fluorescent sensor for glucose

International Nuclear Information System (INIS)

Sun Xiangying; Liu Bin; Jiang Yunbao

2004-01-01

An extremely sensitive monoboronic acid based fluorescent sensor for glucose was developed. This was carried out by assembling a fluorescent monoboronic acid, 3-aminophenylboronic acid (PBA) indirectly onto gold surface via its electrostatic interaction with cysteine (Cys) that was directly assembled on the gold surface. The formation of self-assembled bilayers (SAB) was confirmed and primarily characterized by cyclic voltammetry and X-ray photoelectron spectra (XPS). The SAB containing PBA was found fluorescent and its fluorescence showed an extremely high sensitivity to the presence of glucose and other monosaccharides such as galactose and fructose with quenching constants at 10 8 M -1 order of magnitude compared to those at 10 2 M -1 in bulk solutions. The quenching constants were found to vary in the order of D-glucose>D-galactose>D-fructose>D-mannose that is different from that in bulk solution which shows the highest binding affinity toward D-fructose and very low sensitivity toward glucose. The reported monoboronic acid based SAB fluorescent sensor showed the highest sensitivity towards glucose with the capacity of detecting saccharides of concentration down to nanomolar level. It was also demonstrated that the fluorescence from PBA/Cys/Au can be easily recovered after each measurement event and therefore also represents a new reusable method for immobilizing reagent in fabricating chemosensors

Effect of glucose infusion on endurance performance after beta-adrenoceptor blocker administration

NARCIS (Netherlands)

van Baak, M.A.; Mooij, J.M.

1994-01-01

Effect of glucose infusion on endurance performance after beta-adrenoceptor blocker administration. Van Baak MA, Mooij JM. Department of Human Biology, University of Limburg, Maastricht, The Netherlands. To investigate the effect of glucose (Glc) infusion on endurance performance after

Modeling the Error of the Medtronic Paradigm Veo Enlite Glucose Sensor.

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Biagi, Lyvia; Ramkissoon, Charrise M; Facchinetti, Andrea; Leal, Yenny; Vehi, Josep

2017-06-12

Continuous glucose monitors (CGMs) are prone to inaccuracy due to time lags, sensor drift, calibration errors, and measurement noise. The aim of this study is to derive the model of the error of the second generation Medtronic Paradigm Veo Enlite (ENL) sensor and compare it with the Dexcom SEVEN PLUS (7P), G4 PLATINUM (G4P), and advanced G4 for Artificial Pancreas studies (G4AP) systems. An enhanced methodology to a previously employed technique was utilized to dissect the sensor error into several components. The dataset used included 37 inpatient sessions in 10 subjects with type 1 diabetes (T1D), in which CGMs were worn in parallel and blood glucose (BG) samples were analyzed every 15 ± 5 min Calibration error and sensor drift of the ENL sensor was best described by a linear relationship related to the gain and offset. The mean time lag estimated by the model is 9.4 ± 6.5 min. The overall average mean absolute relative difference (MARD) of the ENL sensor was 11.68 ± 5.07% Calibration error had the highest contribution to total error in the ENL sensor. This was also reported in the 7P, G4P, and G4AP. The model of the ENL sensor error will be useful to test the in silico performance of CGM-based applications, i.e., the artificial pancreas, employing this kind of sensor.

O-GlcNAcase Fragment Discovery with Fluorescence Polarimetry.

Science.gov (United States)

Borodkin, Vladimir S; Rafie, Karim; Selvan, Nithya; Aristotelous, Tonia; Navratilova, Iva; Ferenbach, Andrew T; van Aalten, Daan M F

2018-05-18

The attachment of the sugar N-acetyl-D-glucosamine (GlcNAc) to specific serine and threonine residues on proteins is referred to as protein O-GlcNAcylation. O-GlcNAc transferase (OGT) is the enzyme responsible for carrying out the modification, while O-GlcNAcase (OGA) reverses it. Protein O-GlcNAcylation has been implicated in a wide range of cellular processes including transcription, proteostasis, and stress response. Dysregulation of O-GlcNAc has been linked to diabetes, cancer, and neurodegenerative and cardiovascular disease. OGA has been proposed to be a drug target for the treatment of Alzheimer's and cardiovascular disease given that increased O-GlcNAc levels appear to exert a protective effect. The search for specific, potent, and drug-like OGA inhibitors with bioavailability in the brain is therefore a field of active research, requiring orthogonal high-throughput assay platforms. Here, we describe the synthesis of a novel probe for use in a fluorescence polarization based assay for the discovery of inhibitors of OGA. We show that the probe is suitable for use with both human OGA, as well as the orthologous bacterial counterpart from Clostridium perfringens, CpOGA, and the lysosomal hexosaminidases HexA/B. We structurally characterize CpOGA in complex with a ligand identified from a fragment library screen using this assay. The versatile synthesis procedure could be adapted for making fluorescent probes for the assay of other glycoside hydrolases.

O-GlcNAc modification: why so intimately associated with phosphorylation?

Directory of Open Access Journals (Sweden)

Ande Sudharsana R

2011-01-01

Full Text Available Abstract Post-translational modification of proteins at serine and threonine side chains by β-N-acetylglucosamine (O-GlcNAc mediated by the enzyme β-N-acetylglucosamine transferase has been emerging as a fundamental regulatory mechanism encompassing a wide range of proteins involved in cell division, metabolism, transcription and cell signaling. Furthermore, an extensive interplay between O-GlcNAc modification and serine/threonine phosphorylation in a variety of proteins has been reported to exist. However, our understanding of the regulatory mechanisms involved in O-GlcNAc modification and its interplay with serine/threonine phosphorylation in proteins is still elusive. Recent success in the mapping of O-GlcNAc modification sites in proteins as a result of technological advancement in mass spectrometry have revealed two important clues which may be inherently connected to the regulation of O-GlcNAc modification and its interplay with phosphorylation in proteins. First, almost all O-GlcNAc modified proteins are known phospho proteins. Second, the prevalence of tyrosine phosphorylation among O-GlcNAc modified proteins is exceptionally higher (~68% than its normal occurrence (~2% alone. We hypothesize that phosphorylation may be a requisite for O-GlcNAc modification and tyrosine phosphorylation plays a role in the interplay between O-GlcNAc modification and serine/threonine phosphorylation in proteins. In other words, the interplay between O-GlcNAc modification and phosphorylation is not limited to serine/threonine phosphorylation but also includes tyrosine phosphorylation. Our hypothesis provides an opportunity to understand the underlying mechanism involved in O-GlcNAc modification and its interplay with serine/threonine phosphorylation in proteins. Furthermore, implication of our hypothesis extends to tyrosine kinase signaling.

Enzyme-Free Electrochemical Glucose Sensors Prepared by Dealloying Pd-Ni-P Metallic Glasses

Directory of Open Access Journals (Sweden)

Yuqiao Zeng

2014-01-01

Full Text Available We report the formation of enzyme-free electrochemical glucose sensors by electrochemical dealloying palladium-containing Pd-Ni-P metallic glasses. When metallic glasses with different Pd contents are used as the dealloying precursor alloys, palladium-based nanoporous metals with different ligament and pore sizes can be obtained. The chemical compositions of the nanoporous metals also vary according to the different precursor compositions. All the as-obtained nanoporous metals exhibit electrochemical catalytic activity towards the oxidation of d-glucose, indicating that the nanoporous metals prepared by dealloying the Pd-Ni-P metallic glasses are promising materials for enzyme-free electrochemical glucose sensor.

The emerging link between O-GlcNAcylation and neurological disorders.

Science.gov (United States)

Ma, Xiaofeng; Li, He; He, Yating; Hao, Junwei

2017-10-01

O-linked β-N-acetylglucosaminylation (O-GlcNAcylation) is involved in the regulation of many cellular cascades and neurological diseases such as Alzheimer's disease (AD), Parkinson's disease (PD), and stroke. In the brain, the expression of O-GlcNAcylation is notably heightened, as is that of O-linked N-acetylglucosaminyltransferase (OGT) and β-N-acetylglucosaminidase (OGA), the presence of which is prominent in many regions of neurological importance. Most importantly, O-GlcNAcylation is believed to contribute to the normal functioning of neurons; conversely, its dysregulation participates in the pathogenesis of neurological disorders. In neurodegenerative diseases, O-GlcNAcylation of the brain's key proteins, such as tau and amyloid-β, interacts with their phosphorylation, thereby triggering the formation of neurofibrillary tangles and amyloid plaques. An increase of O-GlcNAcylation by pharmacological intervention prevents neuronal loss. Additionally, O-GlcNAcylation is stress sensitive, and its elevation is cytoprotective. Increased O-GlcNAcylation ameliorated brain damage in victims of both trauma-hemorrhage and stroke. In this review, we summarize the current understanding of O-GlcNAcylation's physiological and pathological roles in the nervous system and provide a foundation for development of a therapeutic strategy for neurological disorders.

A selective glucose sensor based on direct oxidation on a bimetal catalyst with a molecular imprinted polymer.

Science.gov (United States)

Cho, Seong Je; Noh, Hui-Bog; Won, Mi-Sook; Cho, Chul-Ho; Kim, Kwang Bok; Shim, Yoon-Bo

2018-01-15

A selective nonenzymatic glucose sensor was developed based on the direct oxidation of glucose on hierarchical CuCo bimetal-coated with a glucose-imprinted polymer (GIP). Glucose was introduced into the GIP composed of Nafion and polyurethane along with aminophenyl boronic acid (APBA), which was formed on the bimetal electrode formed on a screen-printed electrode. The extraction of glucose from the GIP allowed for the selective permeation of glucose into the bimetal electrode surface for oxidation. The GIP-coated bimetal sensor probe was characterized using electrochemical and surface analytical methods. The GIP layer coated on the NaOH pre-treated bimetal electrode exhibited a dynamic range between 1.0µM and 25.0mM with a detection limit of 0.65±0.10µM in phosphate buffer solution (pH 7.4). The anodic responses of uric acid, acetaminophen, dopamine, ascorbic acid, L-cysteine, and other saccharides (monosaccharides: galactose, mannose, fructose, and xylose; disaccharides: sucrose, lactose, and maltose) were not detected using the GIP-coated bimetal sensor. The reliability of the sensor was evaluated by the determination of glucose in artificial and whole blood samples. Copyright © 2017 Elsevier B.V. All rights reserved.

O-GlcNAcylation regulates ischemia-induced neuronal apoptosis through AKT signaling.

Science.gov (United States)

Shi, Jianhua; Gu, Jin-hua; Dai, Chun-ling; Gu, Jianlan; Jin, Xiaoxia; Sun, Jianming; Iqbal, Khalid; Liu, Fei; Gong, Cheng-Xin

2015-09-28

Apoptosis plays an important role in neural development and neurological disorders. In this study, we found that O-GlcNAcylation, a unique protein posttranslational modification with O-linked β-N-acetylglucosamine (GlcNAc), promoted apoptosis through attenuating phosphorylation/activation of AKT and Bad. By using co-immunoprecipitation and mutagenesis techniques, we identified O-GlcNAc modification at both Thr308 and Ser473 of AKT. O-GlcNAcylation-induced apoptosis was attenuated by over-expression of AKT. We also found a dynamic elevation of protein O-GlcNAcylation during the first four hours of cerebral ischemia, followed by continuous decline after middle cerebral artery occlusion (MCAO) in the mouse brain. The elevation of O-GlcNAcylation coincided with activation of cell apoptosis. Finally, we found a negative correlation between AKT phosphorylation and O-GlcNAcylation in ischemic brain tissue. These results indicate that cerebral ischemia induces a rapid increase of O-GlcNAcylation that promotes apoptosis through down-regulation of AKT activity. These findings provide a novel mechanism through which O-GlcNAcylation regulates ischemia-induced neuronal apoptosis through AKT signaling.

Transcriptional and metabolic effects of glucose on Streptococcus pneumoniae sugar metabolism

Directory of Open Access Journals (Sweden)

Laura ePaixão

2015-10-01

Full Text Available Streptococcus pneumoniae is a strictly fermentative human pathogen that relies on carbohydrate metabolism to generate energy for growth. The nasopharynx colonised by the bacterium is poor in free sugars, but mucosa lining glycans can provide a source of sugar. In blood and inflamed tissues glucose is the prevailing sugar. As a result during progression from colonisation to disease S. pneumoniae has to cope with a pronounced shift in carbohydrate nature and availability. Thus, we set out to assess the pneumococcal response to sugars found in glycans and the influence of glucose (Glc on this response at the transcriptional, physiological and metabolic levels. Galactose (Gal, N-acetylglucosamine (GlcNAc and mannose (Man affected the expression of 8 to 14% of the genes covering cellular functions including central carbon metabolism and virulence. The pattern of end-products as monitored by in vivo 13C-NMR is in good agreement with the fermentation profiles during growth, while the pools of phosphorylated metabolites are consistent with the type of fermentation observed (homolactic vs. mixed and regulation at the metabolic level. Furthermore, the accumulation of α-Gal6P and Man6P indicate metabolic bottlenecks in the metabolism of Gal and Man, respectively. Glc added to cells actively metabolizing other sugar(s was readily consumed and elicited a metabolic shift towards a homolactic profile. The transcriptional response to Glc was large (over 5% of the genome. In central carbon metabolism (most represented category, Glc exerted mostly negative regulation. The smallest response to Glc was observed on a sugar mix, suggesting that exposure to varied sugars improves the fitness of S. pneumoniae. The expression of virulence factors was negatively controlled by Glc in a sugar-dependent manner. Overall, our results shed new light on the link between carbohydrate metabolism, adaptation to host niches and virulence.

Transcriptional and metabolic effects of glucose on Streptococcus pneumoniae sugar metabolism.

Science.gov (United States)

Paixão, Laura; Caldas, José; Kloosterman, Tomas G; Kuipers, Oscar P; Vinga, Susana; Neves, Ana R

2015-01-01

Streptococcus pneumoniae is a strictly fermentative human pathogen that relies on carbohydrate metabolism to generate energy for growth. The nasopharynx colonized by the bacterium is poor in free sugars, but mucosa lining glycans can provide a source of sugar. In blood and inflamed tissues glucose is the prevailing sugar. As a result during progression from colonization to disease S. pneumoniae has to cope with a pronounced shift in carbohydrate nature and availability. Thus, we set out to assess the pneumococcal response to sugars found in glycans and the influence of glucose (Glc) on this response at the transcriptional, physiological, and metabolic levels. Galactose (Gal), N-acetylglucosamine (GlcNAc), and mannose (Man) affected the expression of 8 to 14% of the genes covering cellular functions including central carbon metabolism and virulence. The pattern of end-products as monitored by in vivo (13)C-NMR is in good agreement with the fermentation profiles during growth, while the pools of phosphorylated metabolites are consistent with the type of fermentation observed (homolactic vs. mixed) and regulation at the metabolic level. Furthermore, the accumulation of α-Gal6P and Man6P indicate metabolic bottlenecks in the metabolism of Gal and Man, respectively. Glc added to cells actively metabolizing other sugar(s) was readily consumed and elicited a metabolic shift toward a homolactic profile. The transcriptional response to Glc was large (over 5% of the genome). In central carbon metabolism (most represented category), Glc exerted mostly negative regulation. The smallest response to Glc was observed on a sugar mix, suggesting that exposure to varied sugars improves the fitness of S. pneumoniae. The expression of virulence factors was negatively controlled by Glc in a sugar-dependent manner. Overall, our results shed new light on the link between carbohydrate metabolism, adaptation to host niches and virulence.

Engineering glucose oxidase to minimize the influence of oxygen on sensor response

International Nuclear Information System (INIS)

Horaguchi, Yohei; Saito, Shoko; Kojima, Katsuhiro; Tsugawa, Wakako; Ferri, Stefano; Sode, Koji

2014-01-01

Glucose oxidase (GOx) is an important industrial enzyme and is recognized as the gold standard for monitoring blood glucose. However, due to its inherent oxidase property, the presence of oxygen affects electrochemical measurements of venous blood glucose employing artificial electron mediators. We therefore attempted to engineer Penicillium amagasakiense-derived GOx into a dehydrogenase by focusing on the amino acid residues predicted to interact with oxygen. Our rational amino acid substitution approach resulted in the construction of the Ser114Ala/Phe355Leu mutant, which has an 11-fold decrease in oxidase activity and 2.8-fold increase in dehydrogenase activity compared with wild-type GOx. As a result, the dehydrogenase/oxidase activity ratio of the engineered enzyme was 32-fold greater than that of the wild-type enzyme. The enzyme sensor constructed with Ser114Ala/Phe355Leu was considerably less affected by oxygen than the wild-type GOx-based sensor at lower glucose concentrations

Preparation of Glucose Sensor Using Polydimethylsiloxane / Polypyrrole Complex

Science.gov (United States)

Yasuzawa, Mikito; Inoue, Shigeru; Imai, Shinji

New glucose oxidase (GOD) immobilized glucose sensors were prepared by the electropolymerization of 1-(6-D-gluconamidohexyl) pyrrole (GHP) on the platinum wire electrode precoated with the mixture solution of pyrrole derivative GHP, polydimethylsiloxane (PDS) and Nafion. The addition of Nafion into the precoating mixture solution was essential to obtain suitable sensor sensitivity. However, the sensitivity was about the half of that of the electrode without PDS precoating. Although, the introduction of Nafion was effective to improve the long-term stability of the enzyme-immobilized electrode, the electrode prepared using Nafion, PDS and GHP performed excellent long-term stability even at the measurement and storage temperatures of 40°C. Relatively constant response current was obtained over 30 days under the condition of 40°C and over 9 months measured at 25°C. Moreover, the GOD-immobilized GHP polymer film prepared on the electrode precoated with GHP, PDS and Nafion solution, was found to have excellent hemocompatibility from the result of platelet rich plasma contacting test.

Assessing sensor accuracy for non-adjunct use of continuous glucose monitoring.

Science.gov (United States)

Kovatchev, Boris P; Patek, Stephen D; Ortiz, Edward Andrew; Breton, Marc D

2015-03-01

The level of continuous glucose monitoring (CGM) accuracy needed for insulin dosing using sensor values (i.e., the level of accuracy permitting non-adjunct CGM use) is a topic of ongoing debate. Assessment of this level in clinical experiments is virtually impossible because the magnitude of CGM errors cannot be manipulated and related prospectively to clinical outcomes. A combination of archival data (parallel CGM, insulin pump, self-monitoring of blood glucose [SMBG] records, and meals for 56 pump users with type 1 diabetes) and in silico experiments was used to "replay" real-life treatment scenarios and relate sensor error to glycemic outcomes. Nominal blood glucose (BG) traces were extracted using a mathematical model, yielding 2,082 BG segments each initiated by insulin bolus and confirmed by SMBG. These segments were replayed at seven sensor accuracy levels (mean absolute relative differences [MARDs] of 3-22%) testing six scenarios: insulin dosing using sensor values, threshold, and predictive alarms, each without or with considering CGM trend arrows. In all six scenarios, the occurrence of hypoglycemia (frequency of BG levels ≤50 mg/dL and BG levels ≤39 mg/dL) increased with sensor error, displaying an abrupt slope change at MARD =10%. Similarly, hyperglycemia (frequency of BG levels ≥250 mg/dL and BG levels ≥400 mg/dL) increased and displayed an abrupt slope change at MARD=10%. When added to insulin dosing decisions, information from CGM trend arrows, threshold, and predictive alarms resulted in improvement in average glycemia by 1.86, 8.17, and 8.88 mg/dL, respectively. Using CGM for insulin dosing decisions is feasible below a certain level of sensor error, estimated in silico at MARD=10%. In our experiments, further accuracy improvement did not contribute substantively to better glycemic outcomes.

An NFC-Enabled CMOS IC for a Wireless Fully Implantable Glucose Sensor.

Science.gov (United States)

DeHennis, Andrew; Getzlaff, Stefan; Grice, David; Mailand, Marko

2016-01-01

This paper presents an integrated circuit (IC) that merges integrated optical and temperature transducers, optical interface circuitry, and a near-field communication (NFC)-enabled digital, wireless readout for a fully passive implantable sensor platform to measure glucose in people with diabetes. A flip-chip mounted LED and monolithically integrated photodiodes serve as the transduction front-end to enable fluorescence readout. A wide-range programmable transimpedance amplifier adapts the sensor signals to the input of an 11-bit analog-to-digital converter digitizing the measurements. Measurement readout is enabled by means of wireless backscatter modulation to a remote NFC reader. The system is able to resolve current levels of less than 10 pA with a single fluorescent measurement energy consumption of less than 1 μJ. The wireless IC is fabricated in a 0.6-μm-CMOS process and utilizes a 13.56-MHz-based ISO15693 for passive wireless readout through a NFC interface. The IC is utilized as the core interface to a fluorescent, glucose transducer to enable a fully implantable sensor-based continuous glucose monitoring system.

Digitonin abolishes free 2-deoxy-D-glucose accumulation in isolated rat adipocytes

International Nuclear Information System (INIS)

Thompson, K.; Kleinzeller, A.

1986-01-01

The hypothesis that accumulation against sizable chemical gradients of free (non-phosphorylated) 2-deoxy-D-glucose (2dGlc) in isolated rat adipocytes results from an intracellular compartmentation of free hexose was investigated. Cells exposed to 20 μg/ml digitonin for 10' demonstrated an increased plasma membrane permeability indexed by increased L-glucose entry rates and cellular (presumably cytosolic) protein and K + loss. Functional integrity of intracellular organelles was indicated by the ability of the cells to support ATP-driven 45 Ca 2+ -uptake. Equilibrium 3-O-methylglucose (3-O-MG, a non-accumulated hexose) levels were unaffected. These data suggest a specific permeabilizing action of digitonin at the plasma membrane having no effect on intracellular organelles or passively distributed solutes. Upon addition of digitonin, free 2dGlc fell from 66.5 +/- 8.9 to 7.4 +/- 2.3 pmol/10 5 cells, a value not significantly different from 3-O-MG levels. The gradient of 2dGlc-phosphate was also abolished, as was the increased steady-state free 2dGlc levels induced by insulin. The data argue against a compartmentation model as either the mechanism of adipocyte sugar accumulation or the basis of the steady-state free 2dGlc increase seen with insulin and suggest that an intact plasma membrane is essential to the process

Metabolic fate of glucose in rats with traumatic brain injury and pyruvate or glucose treatments: A NMR spectroscopy study.

Science.gov (United States)

Shijo, Katsunori; Sutton, Richard L; Ghavim, Sima S; Harris, Neil G; Bartnik-Olson, Brenda L

2017-01-01

Administration of sodium pyruvate (SP; 9.08 μmol/kg, i.p.), ethyl pyruvate (EP; 0.34 μmol/kg, i.p.) or glucose (GLC; 11.1 μmol/kg, i.p.) to rats after unilateral controlled cortical impact (CCI) injury has been reported to reduce neuronal loss and improve cerebral metabolism. In the present study these doses of each fuel or 8% saline (SAL; 5.47 nmoles/kg) were administered immediately and at 1, 3, 6 and 23 h post-CCI. At 24 h all CCI groups and non-treated Sham injury controls were infused with [1,2 13 C] glucose for 68 min 13 C nuclear magnetic resonance (NMR) spectra were obtained from cortex + hippocampus tissues from left (injured) and right (contralateral) hemispheres. All three fuels increased lactate labeling to a similar degree in the injured hemisphere. The amount of lactate labeled via the pentose phosphate and pyruvate recycling (PPP + PR) pathway increased in CCI-SAL and was not improved by SP, EP, and GLC treatments. Oxidative metabolism, as assessed by glutamate labeling, was reduced in CCI-SAL animals. The greatest improvement in oxidative metabolism was observed in animals treated with SP and fewer improvements after EP or GLC treatments. Compared to SAL, all three fuels restored glutamate and glutamine labeling via pyruvate carboxylase (PC), suggesting improved astrocyte metabolism following fuel treatment. Only SP treatments restored the amount of [4 13 C] glutamate labeled by the PPP + PR pathway to sham levels. Milder injury effects in the contralateral hemisphere appear normalized by either SP or EP treatments, as increases in the total pool of 13 C lactate and labeling of lactate in glycolysis, or decreases in the ratio of PC/PDH labeling of glutamine, were found only for CCI-SAL and CCI-GLC groups compared to Sham. The doses of SP, EP and GLC examined in this study all enhanced lactate labeling and restored astrocyte-specific PC activity but differentially affected neuronal metabolism after CCI injury. The restoration of

O-GlcNAc regulates NEDD4-1 stability via caspase-mediated pathway

International Nuclear Information System (INIS)

Jiang, Kuan; Bai, Bingyang; Ta, Yajie; Zhang, Tingling; Xiao, Zikang; Wang, Peng George; Zhang, Lianwen

2016-01-01

O-GlcNAc modification of cytosolic and nuclear proteins regulates essential cellular processes such as stress responses, transcription, translation, and protein degradation. Emerging evidence indicates O-GlcNAcylation has a dynamic interplay with ubiquitination in cellular regulation. Here, we report that O-GlcNAc indirectly targets a vital E3 ubiquitin ligase enzyme of NEDD4-1. The protein level of NEDD4-1 is accordingly decreased following an increase of overall O-GlcNAc level upon PUGNAc or glucosamine stimulation. O-GlcNAc transferase (OGT) knockdown, overexpression and mutation results confirm that the stability of NEDD4-1 is negatively regulated by cellular O-GlcNAc. Moreover, the NEDD4-1 degradation induced by PUGNAc or GlcN is significantly inhibited by the caspase inhibitor. Our study reveals a regulation mechanism of NEDD4-1 stability by O-GlcNAcylation. - Highlights: • Reduced NEDD4-1 correlates with increased overall O-GlcNAc level. • OGT negatively regulates NEDD4-1 stability. • O-GlcNAc regulates NEDD4-1 through caspase-mediated pathway.

Ratiometric glucose sensing based on fluorescent oxygen films and glucose oxidase

Directory of Open Access Journals (Sweden)

Fengyu Su

2017-06-01

Full Text Available A new two-layer sensor film was constructed for sensing glucose based on glucose oxidase and oxygen sensing material. The first layer of film containing the oxygen sensor and intra-reference material was polymerized, then the second layer of glucose oxidase and glutaraldehyde was formed on the oxygen sensor layer. The two-layer sensor film has a resolution up to 0.05 mM and a detection range from 0 to 5 mM to glucose. The effects of pH and temperature on the sensing performance were systematically investigated. The selective detection of glucose among other monosaccharides, such as fructose, mannose and galactose indicated that the sensing film has excellent selectivity. The prepared sensor was successfully applied for glucose sample detection of glucose concentration in artificial tears. Keywords: Glucose sensor, Glucose oxidase, Fluorescence, Oxygen film, Diabetes

Bacteria-Templated NiO Nanoparticles/Microstructure for an Enzymeless Glucose Sensor

Directory of Open Access Journals (Sweden)

Settu Vaidyanathan

2016-07-01

Full Text Available The bacterial-induced hollow cylinder NiO (HCNiO nanomaterial was utilized for the enzymeless (without GOx detection of glucose in basic conditions. The determination of glucose in 0.05 M NaOH solution with high sensitivity was performed using cyclic voltammetry (CV and amperometry (i–t. The fundamental electrochemical parameters were analyzed and the obtained values of diffusion coefficient (D, heterogeneous rate constant (ks, electroactive surface coverage (Г, and transfer coefficient (alpha-α are 1.75 × 10−6 cm2/s, 57.65 M−1·s−1, 1.45 × 10−10 mol/cm2, and 0.52 respectively. The peak current of the i–t method shows two dynamic linear ranges of calibration curves 0.2 to 3.5 µM and 0.5 to 250 µM for the glucose electro-oxidation. The Ni2+/Ni3+ couple with the HCNiO electrode and the electrocatalytic properties were found to be sensitive to the glucose oxidation. The green chemistry of NiO preparation from bacteria and the high catalytic ability of the oxyhydroxide (NiOOH is the good choice for the development of a glucose sensor. The best obtained sensitivity and limit of detection (LOD for this sensor were 3978.9 µA mM−1·cm−2 and 0.9 µM, respectively.

Construction of a non-enzymatic sensor based on the poly(o-phenylenediamine)/Ag-NPs composites for detecting glucose in blood

Energy Technology Data Exchange (ETDEWEB)

Wang, Jinxiang; Wang, Meirong [College of Chemistry and Chemical Engineering, Jiangsu Key Laboratory of Environmental Engineering and Monitoring, Yangzhou University, 180 Si–Wang–Ting Road, Yangzhou 225002 (China); Guan, Jun [Clinical Medical College of Yangzhou University, Subei People' s Hospital of Jiangsu Province, Yangzhou 225002 (China); Wang, Chengyin, E-mail: wangcy@yzu.edu.cn [College of Chemistry and Chemical Engineering, Jiangsu Key Laboratory of Environmental Engineering and Monitoring, Yangzhou University, 180 Si–Wang–Ting Road, Yangzhou 225002 (China); Wang, Guoxiu [School of Mathematical and Physical Sciences, University of Technology Sydney, City Campus, Broadway, Sydney, NSW 2007 (Australia)

2017-02-01

A non-enzymatic glucose sensor, based on the silver nanoparticles (Ag-NPs)/poly (o-phenylenediamine) (PoPD) composites, is developed by the electrochemical polymerization of o-phenylenediamine and electrodeposition of silver nanoparticles on an indium tin oxide electrode. The Ag-NPs/PoPD composites are characterized by atomic force microscopy, scanning electronic microscopy and energy dispersive spectrometer. Under the optimized experimental conditions, the proposed glucose sensor demonstrates a wide linear range from 0.15 to 13 mmol L{sup −1} with a correlation coefficient of 0.998. The proposed glucose sensor can be used to detect glucose in blood sample with a satisfactory result. In addition, the proposed sensor presents the advantages, such as facile preparation, low cost, high sensitivity and fast response time. It also exhibits good anti-interference performance and stability. - Highlights: • A facile AgNPs/PoPD/ITO modified sensor was developed for the first time. • The non-enzymatic sensor can detect glucose in human blood directly with a wide detection range. • This sensor is of rapid response, low cost, high sensitivity, and long-time stability.

Superior antibacterial activity of GlcN-AuNP-GO by ultraviolet irradiation

Energy Technology Data Exchange (ETDEWEB)

Govindaraju, Saravanan; Samal, Monica; Yun, Kyusik, E-mail: ykyusik@gachon.ac.kr

2016-12-01

A complete bacterialysis analysis of glucosamine-gold nanoparticle-graphene oxide (GlcN-AuNP-GO) and UV-irradiated GlcN-AuNP-GO was conducted. Analytical characterization of GlcN-AuNPs, GO and GlcN-AuNP-GO revealed UV-Vis absorbance peak at around 230 and 500 nm. Microscopic characterization of prepared nanomaterials was performed by scanning electron microscope, atomic force microscopy, and high-resolution transmission microscopy. The results confirmed that the GlcN-AuNPs were uniformly decorated on the surface and edges of graphene sheets. In addition, potent antibacterial activity of GlcN-AuNP-GO that was UV irradiated for 10 min and normal GlcN-AuNP-GO was detected, compared to the standard drug kanamycin, against both Gram-negative and positive bacteria. The minimum inhibitory concentration (MIC) and fluorescence intensity spectra results for Escherichia coli and Enterococcus faecalis showed that the UV-irradiated GlcN-AuNP-GO has better antibacterial activity than normal GlcN-AuNP-GO and kanamycin. Morphological changes were detected by AFM after treatment. These results confirmed that GlcN-AuNP-GO is a potent antibacterial agent with good potential for use in manufacturing medical instruments, pharmaceutical industries and in waste water treatment. - Highlights: • Glucosamine-gold nanoaprticle-graphene oxide (GlcN-AuNPs-GO) was synthesized. • Analytical and morphological characterizations were revealed. • UV irradiated GlcN-AuNP-GO has provide better antibacterial activity. • Morphological changes of before and after treating bacterial strains were imaged.

Direct electrochemistry of glucose oxidase immobilized on nanostructured gold thin films and its application to bioelectrochemical glucose sensor

International Nuclear Information System (INIS)

Qiu Cuicui; Wang Xia; Liu Xueying; Hou Shifeng; Ma Houyi

2012-01-01

Highlights: ► Au thin films are formed by electrodeposition and galvanic replacement technology. ► Glucose oxidase is stably immobilized via a simple physical adsorption method. ► The direct electrochemical behavior is obtained on the immobilized glucose oxidase. ► An amperometric sensor of glucose with an excellent sensing capability is achieved. - Abstract: Glucose oxidase (GOx) was stably immobilized via a simple physical adsorption method onto the nanostructured Au thin films fabricated by using electrodeposition and galvanic replacement technology, which provides a facile method to prepare morphology-controllable Au films and also facilitates the preparation and application of enzyme modified electrodes. An obvious advantage of the as-prepared enzyme electrode (denoted as GOx/Au/GCE) is that the nano-Au films provide a favorable microenvironment for GOx and facilitate the electron transfer between the active center of GOx and electrodes. Cyclic voltammetry (CV) results indicate that the immobilized GOx displayed a direct, reversible and surface-confined redox reaction in the phosphate buffer solution. Furthermore, the enzyme modified electrode was used as a glucose bioelectrochemical sensor, exhibiting a linear relationship in the concentration ranges of 2.5–32.5 μmol L −1 and 60–130 μmol L −1 with a detection limit of 0.32 μmol L −1 (S/N = 3) at an applied potential of −0.55 V. Due to the excellent stability, sensitivity and anti-interference ability, the Au thin films are hopeful in the construction of glucose biosensors.

Quantitative proteomics identifies altered O-GlcNAcylation of structural, synaptic and memory-associated proteins in Alzheimer's disease: Brain protein O-GlcNAcylation in Alzheimer's disease

Energy Technology Data Exchange (ETDEWEB)

Wang, Sheng [Biological Sciences Division, Pacific Northwest National Laboratory, Richland WA USA; Yang, Feng [Biological Sciences Division, Pacific Northwest National Laboratory, Richland WA USA; Petyuk, Vladislav A. [Biological Sciences Division, Pacific Northwest National Laboratory, Richland WA USA; Shukla, Anil K. [Biological Sciences Division, Pacific Northwest National Laboratory, Richland WA USA; Monroe, Matthew E. [Biological Sciences Division, Pacific Northwest National Laboratory, Richland WA USA; Gritsenko, Marina A. [Biological Sciences Division, Pacific Northwest National Laboratory, Richland WA USA; Rodland, Karin D. [Biological Sciences Division, Pacific Northwest National Laboratory, Richland WA USA; Smith, Richard D. [Biological Sciences Division, Pacific Northwest National Laboratory, Richland WA USA; Qian, Wei-Jun [Biological Sciences Division, Pacific Northwest National Laboratory, Richland WA USA; Gong, Cheng-Xin [New York State Institute for Basic Research in Developmental Disabilities, Staten Island, New York USA; Liu, Tao [Biological Sciences Division, Pacific Northwest National Laboratory, Richland WA USA

2017-07-28

Protein modification by O-linked beta-N-acetylglucosamine (O-GlcNAc) is emerging as an important factor in the pathogenesis of sporadic Alzheimer’s disease. Herein we report the most comprehensive, quantitative proteomics analysis for protein O-GlcNAcylation in post-mortem human brains with and without Alzheimer’s using isobaric tandem mass tags labeling, chemoenzymatic photocleavage enrichment and liquid chromatography coupled to mass spectrometry. A total of 1,850 O-GlcNAc peptides covering 1,094 O-GlcNAcylation sites were identified from 530 proteins in the human brain. 128 O-GlcNAc peptides covering 78 proteins were altered significantly in Alzheimer’s brain as compared to controls (q<0.05). Moreover, alteration of the O-GlcNAc peptide abundance could be attributed more to O-GlcNAcylation level than to protein level changes. The altered O-GlcNAcylated proteins belong to several structural and functional categories, including synaptic proteins, cytoskeleton proteins, and memory-associated proteins. These findings suggest that dysregulation of O-GlcNAcylation of multiple brain proteins may be involved in the development of sporadic Alzheimer’s disease.

Photoaffinity labeling of the human erythrocyte monosaccharide transporter with an aryl azide derivative of D-glucose

International Nuclear Information System (INIS)

Shanahan, M.F.; Wadzinski, B.E.; Lowndes, J.M.; Ruoho, A.E.

1985-01-01

A photoreactive, radioiodinated derivative of glucose, N-(4-iodoazidosalicyl)-6-amido-6-deoxyglucopyranose (IASA-glc), has been synthesized and used as a photoaffinity label for the human erythrocyte monosaccharide transporter. Photoinactivation and photoinsertion are both light-dependent and result in a marked decrease in the absorption spectra of the compound. When [ 125 I]IASA-glc was photolyzed with erythrocyte ghost membranes, photoinsertion of radiolabel was observed in three major regions, spectrin, band 3, and a protein of 58,000 daltons located in the zone 4.5 region. Of the three regions which were photolabeled, only labeling of polypeptides in the zone 4.5 region was partially blocked by D-glucose. In the non-iodinated form, N-(4-azidosalicyl)-6-amido-6-deoxy-glucopyranose inhibited the labeling of the transporter by [ 125 I]IASA-glc more effectively than D-glucose. The ability to synthesize this [ 125 I]containing photoprobe for the monosaccharide transporter at carrier-free levels offers several new advantages for investigating the structure of this transport protein in the erythrocyte

Structural basis of O-GlcNAc recognition by mammalian 14-3-3 proteins.

Science.gov (United States)

Toleman, Clifford A; Schumacher, Maria A; Yu, Seok-Ho; Zeng, Wenjie; Cox, Nathan J; Smith, Timothy J; Soderblom, Erik J; Wands, Amberlyn M; Kohler, Jennifer J; Boyce, Michael

2018-05-21

O-GlcNAc is an intracellular posttranslational modification that governs myriad cell biological processes and is dysregulated in human diseases. Despite this broad pathophysiological significance, the biochemical effects of most O-GlcNAcylation events remain uncharacterized. One prevalent hypothesis is that O-GlcNAc moieties may be recognized by "reader" proteins to effect downstream signaling. However, no general O-GlcNAc readers have been identified, leaving a considerable gap in the field. To elucidate O-GlcNAc signaling mechanisms, we devised a biochemical screen for candidate O-GlcNAc reader proteins. We identified several human proteins, including 14-3-3 isoforms, that bind O-GlcNAc directly and selectively. We demonstrate that 14-3-3 proteins bind O-GlcNAc moieties in human cells, and we present the structures of 14-3-3β/α and γ bound to glycopeptides, providing biophysical insights into O-GlcNAc-mediated protein-protein interactions. Because 14-3-3 proteins also bind to phospho-serine and phospho-threonine, they may integrate information from O-GlcNAc and O-phosphate signaling pathways to regulate numerous physiological functions.

Functional significance of O-GlcNAc modification in regulating neuronal properties.

Science.gov (United States)

Hwang, Hongik; Rhim, Hyewhon

2018-03-01

Post-translational modifications (PTMs) covalently modify proteins and diversify protein functions. Along with protein phosphorylation, another common PTM is the addition of O-linked β-N-acetylglucosamine (O-GlcNAc) to serine and/or threonine residues. O-GlcNAc modification is similar to phosphorylation in that it occurs to serine and threonine residues and cycles on and off with a similar time scale. However, a striking difference is that the addition and removal of the O-GlcNAc moiety on all substrates are mediated by the two enzymes regardless of proteins, O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA), respectively. O-GlcNAcylation can interact or potentially compete with phosphorylation on serine and threonine residues, and thus serves as an important molecular mechanism to modulate protein functions and activation. However, it has been challenging to address the role of O-GlcNAc modification in regulating protein functions at the molecular level due to the lack of convenient tools to determine the sites and degrees of O-GlcNAcylation. Studies in this field have only begun to expand significantly thanks to the recent advances in detection and manipulation methods such as quantitative proteomics and highly selective small-molecule inhibitors for OGT and OGA. Interestingly, multiple brain regions, especially hippocampus, express high levels of both OGT and OGA, and a number of neuron-specific proteins have been reported to undergo O-GlcNAcylation. This review aims to discuss the recent updates concerning the impacts of O-GlcNAc modification on neuronal functions at multiple levels ranging from intrinsic neuronal properties to synaptic plasticity and animal behaviors. Copyright © 2017 Elsevier Ltd. All rights reserved.

O-GlcNAcylation regulates ischemia-induced neuronal apoptosis through AKT signaling

OpenAIRE

Shi, Jianhua; Gu, Jin-hua; Dai, Chun-ling; Gu, Jianlan; Jin, Xiaoxia; Sun, Jianming; Iqbal, Khalid; Liu, Fei; Gong, Cheng-Xin

2015-01-01

Apoptosis plays an important role in neural development and neurological disorders. In this study, we found that O-GlcNAcylation, a unique protein posttranslational modification with O-linked β-N-acetylglucosamine (GlcNAc), promoted apoptosis through attenuating phosphorylation/activation of AKT and Bad. By using co-immunoprecipitation and mutagenesis techniques, we identified O-GlcNAc modification at both Thr308 and Ser473 of AKT. O-GlcNAcylation-induced apoptosis was attenuated by over-expr...

Amperometric glucose sensor based on enhanced catalytic reduction of oxygen using glucose oxidase adsorbed onto core-shell Fe3O4-silica-Au magnetic nanoparticles

International Nuclear Information System (INIS)

Wang Aijun; Li Yongfang; Li Zhonghua; Feng Jiuju; Sun Yanli; Chen Jianrong

2012-01-01

Monodisperse Fe 3 O 4 magnetic nanoparticles (NPs) were prepared under facile solvothermal conditions and successively functionalized with silica and Au to form core/shell Fe 3 O 4 -silica-Au NPs. Furthermore, the samples were used as matrix to construct a glucose sensor based on glucose oxidase (GOD). The immobilized GOD retained its bioactivity with high protein load of 3.92 × 10 −9 mol·cm −2 , and exhibited a surface-controlled quasi-reversible redox reaction, with a fast heterogeneous electron transfer rate of 7.98 ± 0.6 s −1 . The glucose biosensor showed a broad linear range up to 3.97 mM with high sensitivity of 62.45 μA·mM −1 cm −2 and fast response (less than 5 s). - Graphical abstract: Core-shell structured Fe 3 O 4 -silica-Au nanoparticles were prepared and used as matrix to construct an amperometric glucose sensor based on glucose oxidase, which showed broad linear range, high sensitivity, and fast response. Highlights: ► Synthesis of monodispersed Fe 3 O 4 nanoparticles. ► Fabrication of core/shell Fe 3 O 4 -silica-Au nanoparticles. ► Construction of a novel glucose sensor with wide linear range, high sensitivity and fast response.

A Cuprous Oxide Thin Film Non-Enzymatic Glucose Sensor Using Differential Pulse Voltammetry and Other Voltammetry Methods and a Comparison to Different Thin Film Electrodes on the Detection of Glucose in an Alkaline Solution

Directory of Open Access Journals (Sweden)

Yifan Dai

2018-01-01

Full Text Available A cuprous oxide (Cu2O thin layer served as the base for a non-enzymatic glucose sensor in an alkaline medium, 0.1 NaOH solution, with a linear range of 50–200 mg/dL using differential pulse voltammetry (DPV measurement. An X-ray photoelectron spectroscopy (XPS study confirmed the formation of the cuprous oxide layer on the thin gold film sensor prototype. Quantitative detection of glucose in both phosphate-buffered saline (PBS and undiluted human serum was carried out. Neither ascorbic acid nor uric acid, even at a relatively high concentration level (100 mg/dL in serum, interfered with the glucose detection, demonstrating the excellent selectivity of this non-enzymatic cuprous oxide thin layer-based glucose sensor. Chronoamperometry and single potential amperometric voltammetry were used to verify the measurements obtained by DPV, and the positive results validated that the detection of glucose in a 0.1 M NaOH alkaline medium by DPV measurement was effective. Nickel, platinum, and copper are commonly used metals for non-enzymatic glucose detection. The performance of these metal-based sensors for glucose detection using DPV were also evaluated. The cuprous oxide (Cu2O thin layer-based sensor showed the best sensitivity for glucose detection among the sensors evaluated.

Biostability of an implantable glucose sensor chip

Science.gov (United States)

Fröhlich, M.; Birkholz, M.; Ehwald, K. E.; Kulse, P.; Fursenko, O.; Katzer, J.

2012-12-01

Surface materials of an implantable microelectronic chip intended for medical applications were evaluated with respect to their long-term stability in bio-environments. The sensor chip shall apply in a glucose monitor by operating as a microviscosimeter according to the principle of affinity viscosimetry. A monolithic integration of a microelectromechanical system (MEMS) into the sensor chip was successfully performed in a combined 0.25 μm CMOS/BiCMOS technology. In order to study material durability and biostability of the surfaces, sensor chips were exposed to various in vitro and in vivo tests. Corrosional damage of SiON, SiO2 and TiN surfaces was investigated by optical microscopy, ellipsometry and AFM. The results served for optimizing the Back-end-of-Line (BEoL) stack, from which the MEMS was prepared. Corrosion of metal lines could significantly be reduced by improving the topmost passivation layer. The experiments revealed no visible damage of the actuator or other functionally important MEMS elements. Sensor chips were also exposed to human body fluid for three month by implantation into the abdomen of a volunteer. Only small effects were observed for layer thickness and Ra roughness after explantation. In particular, TiN as used for the actuator beam showed no degradation by biocorrosion. The highest degradation rate of about 50 nm per month was revealed for the SiON passivation layer. These results suggest that the sensor chip may safely operate in subcutaneous tissue for a period of several months.

Biostability of an implantable glucose sensor chip

International Nuclear Information System (INIS)

Fröhlich, M; Ehwald, K E; Kulse, P; Fursenko, O; Katzer, J; Birkholz, M

2012-01-01

Surface materials of an implantable microelectronic chip intended for medical applications were evaluated with respect to their long-term stability in bio-environments. The sensor chip shall apply in a glucose monitor by operating as a microviscosimeter according to the principle of affinity viscosimetry. A monolithic integration of a microelectromechanical system (MEMS) into the sensor chip was successfully performed in a combined 0.25 μm CMOS/BiCMOS technology. In order to study material durability and biostability of the surfaces, sensor chips were exposed to various in vitro and in vivo tests. Corrosional damage of SiON, SiO 2 and TiN surfaces was investigated by optical microscopy, ellipsometry and AFM. The results served for optimizing the Back-end-of-Line (BEoL) stack, from which the MEMS was prepared. Corrosion of metal lines could significantly be reduced by improving the topmost passivation layer. The experiments revealed no visible damage of the actuator or other functionally important MEMS elements. Sensor chips were also exposed to human body fluid for three month by implantation into the abdomen of a volunteer. Only small effects were observed for layer thickness and R a roughness after explantation. In particular, TiN as used for the actuator beam showed no degradation by biocorrosion. The highest degradation rate of about 50 nm per month was revealed for the SiON passivation layer. These results suggest that the sensor chip may safely operate in subcutaneous tissue for a period of several months.

Liquid-Phase Packaging of a Glucose Oxidase Solution with Parylene Direct Encapsulation and an Ultraviolet Curing Adhesive Cover for Glucose Sensors

OpenAIRE

Seiichi Takamatsu; Hisanori Takano; Nguyen Binh-Khiem; Tomoyuki Takahata; Eiji Iwase; Kiyoshi Matsumoto; Isao Shimoyama

2010-01-01

We have developed a package for disposable glucose sensor chips using Parylene encapsulation of a glucose oxidase solution in the liquid phase and a cover structure made of an ultraviolet (UV) curable adhesive. Parylene was directly deposited onto a small volume (1 μL) of glucose oxidase solution through chemical vapor deposition. The cover and reaction chamber were constructed on Parylene film using a UV-curable adhesive and photolithography. The package was processed at room temperature to ...

The UPR reduces glucose metabolism via IRE1 signaling.

Science.gov (United States)

van der Harg, Judith M; van Heest, Jessica C; Bangel, Fabian N; Patiwael, Sanne; van Weering, Jan R T; Scheper, Wiep

2017-04-01

Neurons are highly dependent on glucose. A disturbance in glucose homeostasis therefore poses a severe risk that is counteracted by activation of stress responses to limit damage and restore the energy balance. A major stress response that is activated under conditions of glucose deprivation is the unfolded protein response (UPR) that is aimed to restore proteostasis in the endoplasmic reticulum. The key signaling of the UPR involves the transient activation of a transcriptional program and an overall reduction of protein synthesis. Since the UPR is strategically positioned to sense and integrate metabolic stress signals, it is likely that - apart from its adaptive response to restore proteostasis - it also directly affects metabolic pathways. Here we investigate the direct role of the UPR in glucose homeostasis. O-GlcNAc is a post-translational modification that is highly responsive to glucose fluctuations. We find that UPR activation results in decreased O-GlcNAc modification, in line with reduced glucose metabolism. Our data indicate that UPR activation has no direct impact on the upstream processes in glucose metabolism; glucose transporter expression, glucose uptake and hexokinase activity. In contrast, prolonged UPR activation decreases glycolysis and mitochondrial metabolism. Decreased mitochondrial respiration is not accompanied by apoptosis or a structural change in mitochondria indicating that the reduction in metabolic rate upon UPR activation is a physiological non-apoptotic response. Metabolic decrease is prevented if the IRE1 pathway of the UPR is inhibited. This indicates that activation of IRE1 signaling induces a reduction in glucose metabolism, as part of an adaptive response. Copyright © 2017 Elsevier B.V. All rights reserved.

Glucose administration after traumatic brain injury exerts some benefits and no adverse effects on behavioral and histological outcomes.

Science.gov (United States)

Shijo, Katsunori; Ghavim, Sima; Harris, Neil G; Hovda, David A; Sutton, Richard L

2015-07-21

The impact of hyperglycemia after traumatic brain injury (TBI), and even the administration of glucose-containing solutions to head injured patients, remains controversial. In the current study adult male Sprague-Dawley rats were tested on behavioral tasks and then underwent surgery to induce sham injury or unilateral controlled cortical impact (CCI) injury followed by injections (i.p.) with either a 50% glucose solution (Glc; 2g/kg) or an equivalent volume of either 0.9% or 8% saline (Sal) at 0, 1, 3 and 6h post-injury. The type of saline treatment did not significantly affect any outcome measures, so these data were combined. Rats with CCI had significant deficits in beam-walking traversal time and rating scores (p's beam-walking deficits were not affected by Glc versus Sal treatments. Persistent post-CCI deficits in forelimb contraflexion scores and forelimb tactile placing ability were also not differentially affected by Glc or Sal treatments. However, deficits in latency to retract the right hind limb after limb extension were significantly attenuated in the CCI-Glc group (p < 0.05 versus CCI-Sal). Both CCI groups were significantly impaired in a plus maze test of spatial working memory on days 4, 9 and 14 post-surgery (p < 0.001 versus sham), and there was no effect of Glc versus Sal on this cognitive outcome measure. At 15 days post-surgery the loss of cortical tissue volume (p < 0.001 versus sham) was significantly less in the CCI-Glc group (30.0%; p < 0.05) compared to the CCI-Sal group (35.7%). Counts of surviving hippocampal hilar neurons revealed a significant (~40%) loss ipsilateral to CCI (p < 0.001 versus sham), but neuronal loss in the hippocampus was not different in the CCI-Sal and CCI-Glc groups. Taken together, these results indicate that an early elevation of blood glucose may improve some neurological outcomes and, importantly, the induction of hyperglycemia after isolated TBI did not adversely affect any sensorimotor, cognitive or

Effects of hypo-O-GlcNAcylation on Drosophila development.

Science.gov (United States)

Mariappa, Daniel; Ferenbach, Andrew T; van Aalten, Daan M F

2018-05-11

Post-translational modification of serine/threonine residues in nucleocytoplasmic proteins with GlcNAc ( O -GlcNAcylation) is an essential regulatory mechanism in many cellular processes. In Drosophila , null mutants of the Polycomb gene O -GlcNAc transferase ( OGT ; also known as super sex combs ( sxc )) display homeotic phenotypes. To dissect the requirement for O -GlcNAc signaling in Drosophila development, we used CRISPR/Cas9 gene editing to generate rationally designed sxc catalytically hypomorphic or null point mutants. Of the fertile males derived from embryos injected with the CRISPR/Cas9 reagents, 25% produced progeny carrying precise point mutations with no detectable off-target effects. One of these mutants, the catalytically inactive sxc K872M , was recessive lethal, whereas a second mutant, the hypomorphic sxc H537A , was homozygous viable. We observed that reduced total protein O -GlcNAcylation in the sxc H537A mutant is associated with a wing vein phenotype and temperature-dependent lethality. Genetic interaction between sxc H537A and a null allele of Drosophila host cell factor ( dHcf ), encoding an extensively O -GlcNAcylated transcriptional coactivator, resulted in abnormal scutellar bristle numbers. A similar phenotype was also observed in sxc H537A flies lacking a copy of skuld ( skd ), a Mediator complex gene known to affect scutellar bristle formation. Interestingly, this phenotype was independent of OGT Polycomb function or dHcf downstream targets. In conclusion, the generation of the endogenous OGT hypomorphic mutant sxc H537A enabled us to identify pleiotropic effects of globally reduced protein O -GlcNAc during Drosophila development. The mutants generated and phenotypes observed in this study provide a platform for discovery of OGT substrates that are critical for Drosophila development. © 2018 Mariappa et al.

Detection of glucose in the human brain with 1 H MRS at 7 Tesla.

Science.gov (United States)

Kaiser, Lana G; Hirokazu, Kawaguchi; Fukunaga, Masaki; B Matson, Gerald

2016-12-01

A new method is proposed for noninvasive detection of glucose in vivo using proton MR spectroscopy at 7 Tesla. The proposed method utilizes J-difference editing to uncover the resonance of beta-glucose (β-glc) at 3.23 ppm, which is strongly overlapped with choline. Calculations using the density matrix formalism are used to maximize the signal-to-noise ratio of the β-glc resonance at 3.23 ppm. The calculations are verified using phantom and in vivo data collected at 7 Tesla. The proposed method allows observation of the glucose signal at 3.23 ppm in the human brain spectrum. Additional co-edited resonances of N-acetylaspartylglutamatate and glutathione are also detected in the same experiment. The proposed method does not require carbon ( 13 C)- labeled glucose injections and 13 C hardware; as such, it has a potential to provide valuable information on intrinsic glucose concentration in the human brain in vivo. Magn Reson Med 76:1653-1660, 2016. © 2016 International Society for Magnetic Resonance in Medicine. © 2016 International Society for Magnetic Resonance in Medicine.

SNF3 as high affinity glucose sensor and its function in supporting the viability of Candida glabrata under glucose-limited environment

Directory of Open Access Journals (Sweden)

Tzu Shan eNg

2015-12-01

Full Text Available Candida glabrata is an emerging human fungal pathogen that has efficacious nutrient sensing and responsiveness ability. It can be seen through its ability to thrive in diverse range of nutrient limited-human anatomical sites. Therefore, nutrient sensing particularly glucose sensing is thought to be crucial in contributing to the development and fitness of the pathogen. This study aimed to elucidate the role of SNF3 (Sucrose Non Fermenting 3 as a glucose sensor and its possible role in contributing to the fitness and survivability of C. glabrata in glucose-limited environment. The SNF3 knockout strain was constructed and subjected to different glucose concentrations to evaluate its growth, biofilm formation, amphotericin B susceptibility, ex vivo survivability and effects on the transcriptional profiling of the sugar receptor repressor (SRR pathway-related genes. The SNF3Δ strain showed a retarded growth in low glucose environments (0.01% and 0.1% in both fermentation and respiration-preferred conditions but grew well in high glucose concentration environments (1% and 2%. It was also found to be more susceptible to amphotericin B in low glucose environment (0.1% and macrophage engulfment but showed no difference in the biofilm formation capability. The deletion of SNF3 also resulted in the down-regulation of about half of hexose transporters genes (4 out of 9. Overall, the deletion of SNF3 causes significant reduction in the ability of C. glabrata to sense limited surrounding glucose and consequently disrupts its competency to transport and perform the uptake of this critical nutrient. This study highlighted the role of SNF3 as a high affinity glucose sensor and its role in aiding the survivability of C. glabrata particularly in glucose limited environment.

DNA-dispersed graphene/NiO hybrid materials for highly sensitive non-enzymatic glucose sensor

International Nuclear Information System (INIS)

Lv Wei; Jin Fengmin; Guo Quangui; Yang Quanhong; Kang Feiyu

2012-01-01

Highlights: ► We investigated the potential of GNS/NiO/DNA hybrid used as a nonenzymatic sensor. ► DNA is a highly efficient disperse agent for GNS/NiO hybrid than ionic surfactants. ► GNS/NiO/DNA hybrid shows fast electron transfer in the electrochemical reaction. ► GNS/NiO/DNA hybrid shows good detection performance towards glucose. - Abstract: We demonstrate graphene nanosheet/NiO hybrids (GNS/NiO) as the active material for high-performance non-enzymatic glucose sensors. Such sensors are fabricated by DNA-dispersed GNS/NiO suspension deposited on glassy carbon electrodes. ss-DNA shows strong dispersing ability for the GNS/NiO hybrid materials resulting in stable water-dispersible GNS/NiO/DNA hybrids with fully separated layers. The GNS/NiO/DNA hybrids show enhanced electron transfer in the electrocatalytic reaction process, and accordingly, such hybrids modified electrodes show good sensing performance towards glucose and are characterized by large detection ranges, short response periods, low detection limit and high sensitivity and stability.

Liquid-Phase Packaging of a Glucose Oxidase Solution with Parylene Direct Encapsulation and an Ultraviolet Curing Adhesive Cover for Glucose Sensors

Directory of Open Access Journals (Sweden)

Seiichi Takamatsu

2010-06-01

Full Text Available We have developed a package for disposable glucose sensor chips using Parylene encapsulation of a glucose oxidase solution in the liquid phase and a cover structure made of an ultraviolet (UV curable adhesive. Parylene was directly deposited onto a small volume (1 μL of glucose oxidase solution through chemical vapor deposition. The cover and reaction chamber were constructed on Parylene film using a UV-curable adhesive and photolithography. The package was processed at room temperature to avoid denaturation of the glucose oxidase. The glucose oxidase solution was encapsulated and unsealed. Glucose sensing was demonstrated using standard amperometric detection at glucose concentrations between 0.1 and 100 mM, which covers the glucose concentration range of diabetic patients. Our proposed Parylene encapsulation and UV-adhesive cover form a liquid phase glucose-oxidase package that has the advantages of room temperature processing and direct liquid encapsulation of a small volume solution without use of conventional solidifying chemicals.

Ultrasensitive binder-free glucose sensors based on the pyrolysis of in situ grown Cu MOF

DEFF Research Database (Denmark)

Zhang, Xuan; Luo, Jiangshui; Tang, Pengyi

2017-01-01

A non-enzymatic glucose sensor based on carbon/Cu composite materials was developed by the in-situ growth and subsequent pyrolysis of metal-organic frameworks (MOFs) on Cu foam. After pyrolysis, SEM, HRTEM and STEM-EELS were employed to clarify the hierarchical Cu@porous carbon electrode. It is f......A non-enzymatic glucose sensor based on carbon/Cu composite materials was developed by the in-situ growth and subsequent pyrolysis of metal-organic frameworks (MOFs) on Cu foam. After pyrolysis, SEM, HRTEM and STEM-EELS were employed to clarify the hierarchical Cu@porous carbon electrode...... matrix electrode displays ultrahigh sensitivity (10.1 mA cm−2 mM−1), low detection limit (sensors....

Evaluating the clinical accuracy of two continuous glucose sensors using continuous glucose-error grid analysis.

Science.gov (United States)

Clarke, William L; Anderson, Stacey; Farhy, Leon; Breton, Marc; Gonder-Frederick, Linda; Cox, Daniel; Kovatchev, Boris

2005-10-01

To compare the clinical accuracy of two different continuous glucose sensors (CGS) during euglycemia and hypoglycemia using continuous glucose-error grid analysis (CG-EGA). FreeStyle Navigator (Abbott Laboratories, Alameda, CA) and MiniMed CGMS (Medtronic, Northridge, CA) CGSs were applied to the abdomens of 16 type 1 diabetic subjects (age 42 +/- 3 years) 12 h before the initiation of the study. Each system was calibrated according to the manufacturer's recommendations. Each subject underwent a hyperinsulinemic-euglycemic clamp (blood glucose goal 110 mg/dl) for 70-210 min followed by a 1-mg.dl(-1).min(-1) controlled reduction in blood glucose toward a nadir of 40 mg/dl. Arterialized blood glucose was determined every 5 min using a Beckman Glucose Analyzer (Fullerton, CA). CGS glucose recordings were matched to the reference blood glucose with 30-s precision, and rates of glucose change were calculated for 5-min intervals. CG-EGA was used to quantify the clinical accuracy of both systems by estimating combined point and rate accuracy of each system in the euglycemic (70-180 mg/dl) and hypoglycemic (<70 mg/dl) ranges. A total of 1,104 data pairs were recorded in the euglycemic range and 250 data pairs in the hypoglycemic range. Overall correlation between CGS and reference glucose was similar for both systems (Navigator, r = 0.84; CGMS, r = 0.79, NS). During euglycemia, both CGS systems had similar clinical accuracy (Navigator zones A + B, 88.8%; CGMS zones A + B, 89.3%, NS). However, during hypoglycemia, the Navigator was significantly more clinically accurate than the CGMS (zones A + B = 82.4 vs. 61.6%, Navigator and CGMS, respectively, P < 0.0005). CG-EGA is a helpful tool for evaluating and comparing the clinical accuracy of CGS systems in different blood glucose ranges. CG-EGA provides accuracy details beyond other methods of evaluation, including correlational analysis and the original EGA.

Mechanism of the self-condensation of GlcNH₂

DEFF Research Database (Denmark)

Jia, Lingyu; Liu, Xingchen; Qiao, Yan

2017-01-01

A combined experimental and computational study on the imidazolium ionic liquid-promoted conversion of d-Glucosamine (GlcNH2) to deoxyfructosazine (DOF) and fructosazine (FZ) was performed. The pathways for the formation of DOF and FZ via self-condensation of GlcNH2 were investigated by in situ13C...... NMR using site-selectively 13C-labeled GlcNH2. The structural characterization of the reactive species by ESI–MS spectrometry combined with NMR analysis of [13C-1]GlcNH2 indicates that the first carbon (C-1) of GlcNH2 maps onto the corresponding ring carbons of the intermediate, called...... dihydrofructosazine, indicates that both pathways are plausible and that the pathway to DOF is thermodynamically more favorable than that to FZ. The theoretical results are consistent with the experimental observations, and therefore, a detailed and reasonable reaction mechanism was proposed...

Ratiometric glucose sensing based on fluorescent oxygen films and glucose oxidase

OpenAIRE

Fengyu Su; Liqiang Zhang; Xiangxing Kong; Fred Lee; Yanqing Tian; Deirdre R. Meldrum

2017-01-01

A new two-layer sensor film was constructed for sensing glucose based on glucose oxidase and oxygen sensing material. The first layer of film containing the oxygen sensor and intra-reference material was polymerized, then the second layer of glucose oxidase and glutaraldehyde was formed on the oxygen sensor layer. The two-layer sensor film has a resolution up to 0.05 mM and a detection range from 0 to 5 mM to glucose. The effects of pH and temperature on the sensing performance were systemati...

Methods for the Detection, Study, and Dynamic Profiling of O-GlcNAc Glycosylation.

Science.gov (United States)

Thompson, John W; Griffin, Matthew E; Hsieh-Wilson, Linda C

2018-01-01

The addition of O-linked β-N-acetylglucosamine (O-GlcNAc) to serine/threonine residues of proteins is a ubiquitous posttranslational modification found in all multicellular organisms. Like phosphorylation, O-GlcNAc glycosylation (O-GlcNAcylation) is inducible and regulates a myriad of physiological and pathological processes. However, understanding the diverse functions of O-GlcNAcylation is often challenging due to the difficulty of detecting and quantifying the modification. Thus, robust methods to study O-GlcNAcylation are essential to elucidate its key roles in the regulation of individual proteins, complex cellular processes, and disease. In this chapter, we describe a set of chemoenzymatic labeling methods to (1) detect O-GlcNAcylation on proteins of interest, (2) monitor changes in both the total levels of O-GlcNAcylation and its stoichiometry on proteins of interest, and (3) enable mapping of O-GlcNAc to specific serine/threonine residues within proteins to facilitate functional studies. First, we outline a procedure for the expression and purification of a multiuse mutant galactosyltransferase enzyme (Y289L GalT). We then describe the use of Y289L GalT to modify O-GlcNAc residues with a functional handle, N-azidoacetylgalactosamine (GalNAz). Finally, we discuss several applications of the copper-catalyzed azide-alkyne cycloaddition "click" reaction to attach various alkyne-containing chemical probes to GalNAz and demonstrate how this functionalization of O-GlcNAc-modified proteins can be used to realize (1)-(3) above. Overall, these methods, which utilize commercially available reagents and standard protein analytical tools, will serve to advance our understanding of the diverse and important functions of O-GlcNAcylation. © 2018 Elsevier Inc. All rights reserved.

The early metazoan Trichoplax adhaerens possesses a functional O-GlcNAc system

NARCIS (Netherlands)

Selvan, Nithya; Mariappa, Daniel; Van Den Toorn, Henk W P; Heck, Albert J R; Ferenbach, Andrew T.; Van Aalten, Daan M F

2015-01-01

Protein O-GlcNAcylation is a reversible post-translational signaling modification of nucleocytoplasmic proteins that is essential for embryonic development in bilateria. In a search for a reductionist model to study O-GlcNAc signaling, we discovered the presence of functional O-GlcNAc transferase

Real-time continuous glucose monitoring shows high accuracy within 6 hours after sensor calibration: a prospective study.

Directory of Open Access Journals (Sweden)

Xiao-Yan Yue

Full Text Available Accurate and timely glucose monitoring is essential in intensive care units. Real-time continuous glucose monitoring system (CGMS has been advocated for many years to improve glycemic management in critically ill patients. In order to determine the effect of calibration time on the accuracy of CGMS, real-time subcutaneous CGMS was used in 18 critically ill patients. CGMS sensor was calibrated with blood glucose measurements by blood gas/glucose analyzer every 12 hours. Venous blood was sampled every 2 to 4 hours, and glucose concentration was measured by standard central laboratory device (CLD and by blood gas/glucose analyzer. With CLD measurement as reference, relative absolute difference (mean±SD in CGMS and blood gas/glucose analyzer were 14.4%±12.2% and 6.5%±6.2%, respectively. The percentage of matched points in Clarke error grid zone A was 74.8% in CGMS, and 98.4% in blood gas/glucose analyzer. The relative absolute difference of CGMS obtained within 6 hours after sensor calibration (8.8%±7.2% was significantly less than that between 6 to 12 hours after calibration (20.1%±13.5%, p<0.0001. The percentage of matched points in Clarke error grid zone A was also significantly higher in data sets within 6 hours after calibration (92.4% versus 57.1%, p<0.0001. In conclusion, real-time subcutaneous CGMS is accurate in glucose monitoring in critically ill patients. CGMS sensor should be calibrated less than 6 hours, no matter what time interval recommended by manufacturer.

Aluminum gallium nitride (GaN)/GaN high electron mobility transistor-based sensors for glucose detection in exhaled breath condensate.

Science.gov (United States)

Chu, Byung Hwan; Kang, Byoung Sam; Hung, Sheng Chun; Chen, Ke Hung; Ren, Fan; Sciullo, Andrew; Gila, Brent P; Pearton, Stephen J

2010-01-01

Immobilized aluminum gallium nitride (AlGaN)/GaN high electron mobility transistors (HEMTs) have shown great potential in the areas of pH, chloride ion, and glucose detection in exhaled breath condensate (EBC). HEMT sensors can be integrated into a wireless data transmission system that allows for remote monitoring. This technology offers the possibility of using AlGaN/GaN HEMTs for extended investigations of airway pathology of detecting glucose in EBC without the need for clinical visits. HEMT structures, consisting of a 3-microm-thick undoped GaN buffer, 30-A-thick Al(0.3)Ga(0.7)N spacer, and 220-A-thick silicon-doped Al(0.3)Ga(0.7)N cap layer, were used for fabricating the HEMT sensors. The gate area of the pH, chloride ion, and glucose detection was immobilized with scandium oxide (Sc(2)O(3)), silver chloride (AgCl) thin film, and zinc oxide (ZnO) nanorods, respectively. The Sc(2)O(3)-gated sensor could detect the pH of solutions ranging from 3 to 10 with a resolution of approximately 0.1 pH. A chloride ion detection limit of 10(-8) M was achieved with a HEMT sensor immobilized with the AgCl thin film. The drain-source current of the ZnO nanorod-gated AlGaN/GaN HEMT sensor immobilized with glucose oxidase showed a rapid response of less than 5 seconds when the sensor was exposed to the target glucose in a buffer with a pH value of 7.4. The sensor could detect a wide range of concentrations from 0.5 nM to 125 microM. There is great promise for using HEMT-based sensors to enhance the detection sensitivity for glucose detection in EBC. Depending on the immobilized material, HEMT-based sensors can be used for sensing different materials. These electronic detection approaches with rapid response and good repeatability show potential for the investigation of airway pathology. The devices can also be integrated into a wireless data transmission system for remote monitoring applications. This sensor technology could use the exhaled breath condensate to measure the

Regulatory O-GlcNAcylation sites on FoxO1 are yet to be identified

Energy Technology Data Exchange (ETDEWEB)

Fardini, Yann [INSERM, U1016, Institut Cochin, Paris (France); CNRS, UMR8104, Paris (France); Université Paris Descartes, Sorbonne Paris Cité, Paris (France); Perez-Cervera, Yobana [Structural and Functional Glycobiology Unit, Lille 1 University, CNRS (UMR 8576), IFR 117, Villeneuve d' Ascq (France); Facultad de Odontología, Universidad Autónoma Benito Juárez de Oaxaca, Oaxaca (Mexico); Camoin, Luc [INSERM, U1068, CRCM, Marseille Protéomique IBiSA, Marseille, F-13009 (France); Institut Paoli-Calmettes Team, Cell Polarity, Cell Signaling and Cancer, Marseille, F-13009 (France); Aix-Marseille Université, F-13284, Marseille (France); CNRS, UMR7258, CRCM, Marseille, F-13009 (France); Pagesy, Patrick [INSERM, U1016, Institut Cochin, Paris (France); CNRS, UMR8104, Paris (France); Université Paris Descartes, Sorbonne Paris Cité, Paris (France); Lefebvre, Tony [Structural and Functional Glycobiology Unit, Lille 1 University, CNRS (UMR 8576), IFR 117, Villeneuve d' Ascq (France); Issad, Tarik, E-mail: tarik.issad@inserm.fr [INSERM, U1016, Institut Cochin, Paris (France); CNRS, UMR8104, Paris (France); Université Paris Descartes, Sorbonne Paris Cité, Paris (France)

2015-06-26

O-GlcNAcylation is a reversible post-translational modification that regulates cytosolic and nuclear proteins. We and others previously demonstrated that FoxO1 is O-GlcNAcylated in different cell types, resulting in an increase in its transcriptional activity. Four O-GlcNAcylation sites were identified in human FOXO1 but directed mutagenesis of each site individually had modest (T317) or no effect (S550, T648, S654) on its O-GlcNAcylation status and transcriptional activity. Moreover, the consequences of mutating all four sites had not been investigated. In the present work, we mutated these sites in the mouse Foxo1 and found that mutation of all four sites did not decrease Foxo1 O-GlcNAcylation status and transcriptional activity, and would even tend to increase them. In an attempt to identify other O-GlcNAcylation sites, we immunoprecipitated wild-type O-GlcNAcylated Foxo1 and analysed the tryptic digest peptides by mass spectrometry using High-energy Collisional Dissociation. We identified T646 as a new O-GlcNAcylation site on Foxo1. However, site directed mutagenesis of this site individually or together with all four previously identified residues did not impair Foxo1 O-GlcNAcylation and transcriptional activity. These results suggest that residues important for the control of Foxo1 activity by O-GlcNAcylation still remain to be identified. - Highlights: • We mutate four previously identified O-GlcNAcylation sites on Foxo1. • Unexpectedly, these mutations do not reduce Foxo1 O-GlcNAcylation. • These mutation do not reduce Foxo1 transcriptional activity. • We identify a new O-GlcNAcylation site on Foxo1 by mass spectrometry. • Mutation of this site increases Foxo1 transcriptional activity.

Regulatory O-GlcNAcylation sites on FoxO1 are yet to be identified

International Nuclear Information System (INIS)

Fardini, Yann; Perez-Cervera, Yobana; Camoin, Luc; Pagesy, Patrick; Lefebvre, Tony; Issad, Tarik

2015-01-01

O-GlcNAcylation is a reversible post-translational modification that regulates cytosolic and nuclear proteins. We and others previously demonstrated that FoxO1 is O-GlcNAcylated in different cell types, resulting in an increase in its transcriptional activity. Four O-GlcNAcylation sites were identified in human FOXO1 but directed mutagenesis of each site individually had modest (T317) or no effect (S550, T648, S654) on its O-GlcNAcylation status and transcriptional activity. Moreover, the consequences of mutating all four sites had not been investigated. In the present work, we mutated these sites in the mouse Foxo1 and found that mutation of all four sites did not decrease Foxo1 O-GlcNAcylation status and transcriptional activity, and would even tend to increase them. In an attempt to identify other O-GlcNAcylation sites, we immunoprecipitated wild-type O-GlcNAcylated Foxo1 and analysed the tryptic digest peptides by mass spectrometry using High-energy Collisional Dissociation. We identified T646 as a new O-GlcNAcylation site on Foxo1. However, site directed mutagenesis of this site individually or together with all four previously identified residues did not impair Foxo1 O-GlcNAcylation and transcriptional activity. These results suggest that residues important for the control of Foxo1 activity by O-GlcNAcylation still remain to be identified. - Highlights: • We mutate four previously identified O-GlcNAcylation sites on Foxo1. • Unexpectedly, these mutations do not reduce Foxo1 O-GlcNAcylation. • These mutation do not reduce Foxo1 transcriptional activity. • We identify a new O-GlcNAcylation site on Foxo1 by mass spectrometry. • Mutation of this site increases Foxo1 transcriptional activity

O-GlcNAc inhibits interaction between Sp1 and Elf-1 transcription factors

International Nuclear Information System (INIS)

Lim, Kihong; Chang, Hyo-Ihl

2009-01-01

The novel protein modification, O-linked N-acetylglucosamine (O-GlcNAc), plays an important role in various aspects of cell regulation. Although most of nuclear transcription regulatory factors are modified by O-GlcNAc, O-GlcNAc effects on transcription remain largely undefined yet. In this study, we show that O-GlcNAc inhibits a physical interaction between Sp1 and Elf-1 transcription factors, and negatively regulates transcription of placenta and embryonic expression oncofetal protein gene (Pem). These findings suggest that O-GlcNAc inhibits Sp1-mediated gene transcription possibly by interrupting Sp1 interaction with its cooperative factor.

Nonenzymatic flexible field-effect transistor based glucose sensor fabricated using NiO quantum dots modified ZnO nanorods.

Science.gov (United States)

Jung, Da-Un-Jin; Ahmad, Rafiq; Hahn, Yoon-Bong

2018-02-15

Herein, we fabricated nonenzymatic flexible field-effect transistor (f-FET) based glucose sensor using nickel oxide quantum dots (NiO QDs) modified zinc oxide nanorods (ZnO NRs). The ZnO NRs surfaces were coated with NiO QDs using radio frequency (RF) magnetron sputtering to enhance the electrocatalytic feature and the surface area of ZnO NRs. Under physiological conditions (pH 7.4), the nonenzymatic f-FET glucose sensor shows two linear ranges of 0.001-10mM and 10-50mM with the high sensitivity of 13.14μAcm -2 mM -1 and 7.31μAcm -2 mM -1 , respectively, along with good selectivity, stability and repeatability during glucose detection. The examination of human whole blood and serum samples reveal that the nonenzymatic f-FET based glucose sensor is capable of measuring glucose concentration efficiently in the presence of interfering species and thus can be offered as a promising device for further applications in clinical and non-clinical fields. Copyright © 2017 Elsevier Inc. All rights reserved.

Autoregressive Modeling of Drift and Random Error to Characterize a Continuous Intravascular Glucose Monitoring Sensor.

Science.gov (United States)

Zhou, Tony; Dickson, Jennifer L; Geoffrey Chase, J

2018-01-01

Continuous glucose monitoring (CGM) devices have been effective in managing diabetes and offer potential benefits for use in the intensive care unit (ICU). Use of CGM devices in the ICU has been limited, primarily due to the higher point accuracy errors over currently used traditional intermittent blood glucose (BG) measures. General models of CGM errors, including drift and random errors, are lacking, but would enable better design of protocols to utilize these devices. This article presents an autoregressive (AR) based modeling method that separately characterizes the drift and random noise of the GlySure CGM sensor (GlySure Limited, Oxfordshire, UK). Clinical sensor data (n = 33) and reference measurements were used to generate 2 AR models to describe sensor drift and noise. These models were used to generate 100 Monte Carlo simulations based on reference blood glucose measurements. These were then compared to the original CGM clinical data using mean absolute relative difference (MARD) and a Trend Compass. The point accuracy MARD was very similar between simulated and clinical data (9.6% vs 9.9%). A Trend Compass was used to assess trend accuracy, and found simulated and clinical sensor profiles were similar (simulated trend index 11.4° vs clinical trend index 10.9°). The model and method accurately represents cohort sensor behavior over patients, providing a general modeling approach to any such sensor by separately characterizing each type of error that can arise in the data. Overall, it enables better protocol design based on accurate expected CGM sensor behavior, as well as enabling the analysis of what level of each type of sensor error would be necessary to obtain desired glycemic control safety and performance with a given protocol.

A glucose-responsive pH-switchable bioelectrocatalytic sensor based on phenylboronic acid-diol specificity

International Nuclear Information System (INIS)

Gao, Peiyi; Wang, Zhihua; Yang, Lele; Ma, Tengfei; Yang, Ling; Guo, Qianqiong; Huang, Shasheng

2015-01-01

Graphical abstract: A pH-switchable bioelectrocatayltic sensor was developed, which exhibited an obvious anodic current in acidic conditions as “ON” state, yet a prohibited signal in alkaline conditions as “OFF” state. With the change of pH and/or the presence of glucose, our proposed biosensor produced the corresponding amplified signal, providing a better sensitivity. - Abstract: Aminophenylboronic acid moieties were covalently grafted onto mercaptobenzoic acid moieties, and glucose oxidase was then immobilized through boronic acid-diol specific recognition to form a pH-sensitive electrosensor switching between pH 5.8 and pH 8.0 base solution. Using potassium ferricyanide as electroactive probe, the response was intensified in acidic condition while hindered in alkaline condition. The sharp and stable contrast in current was performed alternately upon the change of pH like a “pH switch”. In the presence of glucose, the response to glucose was further amplified catalytically by glucose oxidase on “ON” state, while electron transfer was inhibited on “OFF” state. Furthermore, when our sensor was on “ON” state, it showed a good linearity ranging from 0 to 30 μmol L −1 of glucose, with a detection limit of 348 nmol L −1 (S/B = 3) and a dynamic range extending to 50 μmol L −1 . Glucose-responsive, pH-switchable and catalytically-amplified, our biosensor provided a new method for the detection of glucose in the form of pH switch in human serum sample, and was promising to more complicated environment

Synthesis of a Benzene-containing C1-Phosphonate Analogue of UDP-GlcNAc for the Inhibition of O-GlcNAc Transferase

Energy Technology Data Exchange (ETDEWEB)

Im, Jungkyun [Soonchunhyang Univ., Asan (Korea, Republic of)

2016-01-15

I report here the design, synthesis, and biological evaluation of a new C1-phosphonate analogue of UDP-GlcNAc as a potential inhibitor of OGT, an enzyme responsible for O-GlcNAc modification. The analogue was designed to mimic the transition state of the natural donor involved in the enzymatic reaction. However, the analogue showed somehow low activity as an inhibitor of OGT.

Crosstalk between phosphorylation and O-GlcNAcylation: friend or foe.

Science.gov (United States)

van der Laarse, Saar A M; Leney, Aneika C; Heck, Albert J R

2018-05-02

A wide variety of protein post-translational modifications (PTMs) decorate cellular proteins, regulating their structure, interactions and ultimately their function. The density of co-occurring PTMs on proteins can be very high, where multiple PTMs can positively or negatively influence each other's actions, termed PTM crosstalk. In this review, we highlight recent progress in the area of PTM crosstalk, whereby we focus on crosstalk between protein phosphorylation and O-GlcNAcylation. These two PTMs largely target identical (i.e., Ser and Thr) amino acids in proteins. Phosphorylation/O-GlcNAcylation crosstalk comes in many flavors, for instance by competition for the same site/residue (reciprocal crosstalk), as well as by modifications influencing each other in proximity or even distal on the protein sequence. PTM crosstalk is observed on the writers of these modifications (i.e., kinases and O-GlcNAc transferase), on the erasers (i.e., phosphatases and O-GlcNAcase), and on the readers and the substrates. We describe examples of all these different flavors of crosstalk, and additionally the methods that are emerging to better investigate in particular phosphorylation/O-GlcNAcylation crosstalk. © 2018 Federation of European Biochemical Societies.

Amperometric glucose sensor based on enhanced catalytic reduction of oxygen using glucose oxidase adsorbed onto core-shell Fe{sub 3}O{sub 4}-silica-Au magnetic nanoparticles

Energy Technology Data Exchange (ETDEWEB)

Wang Aijun [College of Geography and Environmental Science, College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004 (China); Key Laboratory of Green Chemical Media and Reactions, Ministry of Education, School of Chemistry and Environmental Science, Henan Normal University, Xinxiang 453007 (China); Li Yongfang [College of Chemistry and Chemical Engineering, Henan Institute of Science and Technology, Xinxiang 453003 (China); Li Zhonghua [Key Laboratory of Green Chemical Media and Reactions, Ministry of Education, School of Chemistry and Environmental Science, Henan Normal University, Xinxiang 453007 (China); Feng Jiuju, E-mail: jjfengnju@gmail.com [College of Geography and Environmental Science, College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004 (China); Key Laboratory of Green Chemical Media and Reactions, Ministry of Education, School of Chemistry and Environmental Science, Henan Normal University, Xinxiang 453007 (China); Sun Yanli [Key Laboratory of Green Chemical Media and Reactions, Ministry of Education, School of Chemistry and Environmental Science, Henan Normal University, Xinxiang 453007 (China); Chen Jianrong [College of Geography and Environmental Science, College of Chemistry and Life Science, Zhejiang Normal University, Jinhua 321004 (China)

2012-08-01

Monodisperse Fe{sub 3}O{sub 4} magnetic nanoparticles (NPs) were prepared under facile solvothermal conditions and successively functionalized with silica and Au to form core/shell Fe{sub 3}O{sub 4}-silica-Au NPs. Furthermore, the samples were used as matrix to construct a glucose sensor based on glucose oxidase (GOD). The immobilized GOD retained its bioactivity with high protein load of 3.92 Multiplication-Sign 10{sup -9} mol{center_dot}cm{sup -2}, and exhibited a surface-controlled quasi-reversible redox reaction, with a fast heterogeneous electron transfer rate of 7.98 {+-} 0.6 s{sup -1}. The glucose biosensor showed a broad linear range up to 3.97 mM with high sensitivity of 62.45 {mu}A{center_dot}mM{sup -1} cm{sup -2} and fast response (less than 5 s). - Graphical abstract: Core-shell structured Fe{sub 3}O{sub 4}-silica-Au nanoparticles were prepared and used as matrix to construct an amperometric glucose sensor based on glucose oxidase, which showed broad linear range, high sensitivity, and fast response. Highlights: Black-Right-Pointing-Pointer Synthesis of monodispersed Fe{sub 3}O{sub 4} nanoparticles. Black-Right-Pointing-Pointer Fabrication of core/shell Fe{sub 3}O{sub 4}-silica-Au nanoparticles. Black-Right-Pointing-Pointer Construction of a novel glucose sensor with wide linear range, high sensitivity and fast response.

Non-Enzymatic Glucose Sensor Composed of Carbon-Coated Nano-Zinc Oxide

Directory of Open Access Journals (Sweden)

Ren-Jei Chung

2017-02-01

Full Text Available Nowadays glucose detection is of great importance in the fields of biological, environmental, and clinical analyzes. In this research, we report a zinc oxide (ZnO nanorod powder surface-coated with carbon material for non-enzymatic glucose sensor applications through a hydrothermal process and chemical vapor deposition method. A series of tests, including crystallinity analysis, microstructure observation, and electrochemical property investigations were carried out. For the cyclic voltammetric (CV glucose detection, the low detection limit of 1 mM with a linear range from 0.1 mM to 10 mM was attained. The sensitivity was 2.97 μA/cm2mM, which is the most optimized ever reported. With such good analytical performance from a simple process, it is believed that the nanocomposites composed of ZnO nanorod powder surface-coated with carbon material are promising for the development of cost-effective non-enzymatic electrochemical glucose biosensors with high sensitivity.

The Hsp70 homolog Ssb and the 14-3-3 protein Bmh1 jointly regulate transcription of glucose repressed genes in Saccharomyces cerevisiae.

Science.gov (United States)

Hübscher, Volker; Mudholkar, Kaivalya; Chiabudini, Marco; Fitzke, Edith; Wölfle, Tina; Pfeifer, Dietmar; Drepper, Friedel; Warscheid, Bettina; Rospert, Sabine

2016-07-08

Chaperones of the Hsp70 family interact with a multitude of newly synthesized polypeptides and prevent their aggregation. Saccharomyces cerevisiae cells lacking the Hsp70 homolog Ssb suffer from pleiotropic defects, among others a defect in glucose-repression. The highly conserved heterotrimeric kinase SNF1/AMPK (AMP-activated protein kinase) is required for the release from glucose-repression in yeast and is a key regulator of energy balance also in mammalian cells. When glucose is available the phosphatase Glc7 keeps SNF1 in its inactive, dephosphorylated state. Dephosphorylation depends on Reg1, which mediates targeting of Glc7 to its substrate SNF1. Here we show that the defect in glucose-repression in the absence of Ssb is due to the ability of the chaperone to bridge between the SNF1 and Glc7 complexes. Ssb performs this post-translational function in concert with the 14-3-3 protein Bmh, to which Ssb binds via its very C-terminus. Raising the intracellular concentration of Ssb or Bmh enabled Glc7 to dephosphorylate SNF1 even in the absence of Reg1. By that Ssb and Bmh efficiently suppressed transcriptional deregulation of Δreg1 cells. The findings reveal that Ssb and Bmh comprise a new chaperone module, which is involved in the fine tuning of a phosphorylation-dependent switch between respiration and fermentation. © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

Phenylboronic acid functionalized reduced graphene oxide based fluorescence nano sensor for glucose sensing

Energy Technology Data Exchange (ETDEWEB)

Basiruddin, SK; Swain, Sarat K., E-mail: swainsk2@yahoo.co.in

2016-01-01

Reduced graphene has emerged as promising tools for detection based application of biomolecules as it has high surface area with strong fluorescence quenching property. We have used the concept of fluorescent quenching property of reduced graphene oxide to the fluorescent probes which are close vicinity of its surface. In present work, we have synthesized fluorescent based nano-sensor consist of phenylboronic acid functionalized reduced graphene oxide (rGO–PBA) and di-ol modified fluorescent probe for detection of biologically important glucose molecules. This fluorescent graphene based nano-probe has been characterized by high resolution transmission electron microscope (HRTEM), Atomic force microscope (AFM), UV–visible, Photo-luminescence (PL) and Fourier transformed infrared (FT-IR) spectroscopy. Finally, using this PBA functionalized reduced GO based nano-sensor, we were able to detect glucose molecule in the range of 2 mg/mL to 75 mg/mL in aqueous solution of pH 7.4. - Highlights: • Easy and simple synthesis of PBA functionalized reduced GO based nano probe. • PBA functionalized reduced GO graphene based nano-probes are characterized. • PBA functionalized reduced GO nano probe is used to detect glucose molecules. • It is very cost-effective and enzyme-free detection of glucose in solution.

The interaction between glucose and cytokinin signaling in controlling Arabidopsis thaliana seedling root growth and development.

Science.gov (United States)

Kushwah, Sunita; Laxmi, Ashverya

2017-05-04

Cytokinin (CK) and glucose (GLC) control several common responses in plants. There is an extensive overlap between CK and GLC signal transduction pathways in Arabidopsis. Physiologically, both GLC and CK could regulate root length in light. CK interacts with GLC via HXK1 dependent pathway for root length control. Wild-type (WT) roots cannot elongate in the GLC free medium while CK-receptor mutant ARABIDOPSIS HISTIDINE KINASE4 (ahk4) and type B ARR triple mutant ARABIDOPSIS RESPONSE REGULATOR1, 10,11 (arr1, 10,11) roots could elongate even in the absence of GLC as compared with the WT. The root hair initiation was also found defective in CK signaling mutants ahk4, arr1,10,11 and arr3,4,5,6,8,9 on increasing GLC concentration (up to 3%); and lesser number of root hairs were visible even at 5% GLC as compared with the WT. Out of 941 BAP regulated genes, 103 (11%) genes were involved in root growth and development. Out of these 103 genes, 60 (58%) genes were also regulated by GLC. GLC could regulate 5736 genes, which include 327 (6%) genes involved in root growth and development. Out of these 327 genes, 60 (18%) genes were also regulated by BAP. Both GLC and CK signaling cannot alter root length in light in auxin signaling mutant AUXIN RESPONSE3/INDOLE-3-ACETIC ACID17 (axr3/iaa17) suggesting that they may involve auxin signaling component as a nodal point. Therefore CK- and GLC- signaling are involved in controlling different aspects of root growth and development such as root length, with auxin signaling components working as downstream target.

Hsp70-GlcNAc-binding activity is released by stress, proteasome inhibition, and protein misfolding

International Nuclear Information System (INIS)

Guinez, Celine; Mir, Anne-Marie; Leroy, Yves; Cacan, Rene; Michalski, Jean-Claude; Lefebvre, Tony

2007-01-01

Numerous recent works strengthen the idea that the nuclear and cytosolic-specific O-GlcNAc glycosylation protects cells against injuries. We have first investigated O-GlcNAc level and Hsp70-GlcNAc-binding activity (HGBA) behaviour after exposure of HeLa and HepG 2 cells to a wide variety of stresses. O-GlcNAc and HGBA responses were different according to the stress and according to the cell. HGBA was released for almost all stresses, while O-GlcNAc level was modified either upwards or downwards, depending to the stress. Against all expectations, we demonstrated that energy charge did not significantly vary with stress whereas UDP-GlcNAc pools were more dramatically affected even if differences in UDP-GlcNAc contents were not correlated with O-GlcNAc variations suggesting that O-GlcNAc transferase is itself finely regulated during cell injury. Finally, HGBA could be triggered by proteasome inhibition and by L-azetidine-2-carboxylic acid (a proline analogue) incorporation demonstrating that protein misfolding is one of the key-activator of this Hsp70 property

Efficient 1H-NMR Quantitation and Investigation of N-Acetyl-D-glucosamine (GlcNAc and N,N'-Diacetylchitobiose (GlcNAc2 from Chitin

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Huey-Lang Yang

2011-09-01

Full Text Available A quantitative determination method of N-acetyl-D-glucosamine (GlcNAc and N,N'-diacetylchitobiose (GlcNAc2 is proposed using a proton nuclear magnetic resonance experiment. N-acetyl groups of GlcNAc and (GlcNAc2 are chosen as target signals, and the deconvolution technique is used to determine the concentration of the corresponding compound. Compared to the HPLC method, 1H-NMR spectroscopy is simple and fast. The method can be used for the analysis of chitin hydrolyzed products with real-time analysis, and for quantifying the content of products using internal standards without calibration curves. This method can be used to quickly evaluate chitinase activity. The temperature dependence of 1H-NMR spectra (VT-NMR is studied to monitor the chemical shift variation of acetyl peak. The acetyl groups of products are involved in intramolecular H-bonding with the OH group on anomeric sites. The rotation of the acetyl group is closely related to the intramolecular hydrogen bonding pattern, as suggested by the theoretical data (molecular modeling.

Glucose determination using a re-usable enzyme-modified ion track membrane sensor

Czech Academy of Sciences Publication Activity Database

Fink, Dietmar; Klinkovich, I.; Bukelman, O.; Marks, R.S.; Kiv, A.; Fuks, D.; Fahrner, W. R.; Alfonta, L.

2009-01-01

Roč. 24, č. 8 (2009), s. 2702-2706 ISSN 0956-5663 Institutional research plan: CEZ:AV0Z10480505 Keywords : Glucose sensor * etched tracks * Ion track membranes Subject RIV: BG - Nuclear, Atomic and Molecular Physics, Colliders Impact factor: 5.429, year: 2009

Biostable glucose permeable polymer

DEFF Research Database (Denmark)

2017-01-01

A new biostable glucose permeable polymer has been developed which is useful, for example, in implantable glucose sensors. This biostable glucose permeable polymer has a number of advantageous characteristics and, for example, does not undergo hydrolytic cleavage and degradation, thereby providing...... a composition that facilitates long term sensor stability in vivo. The versatile characteristics of this polymer allow it to be used in a variety of contexts, for example to form the body of an implantable glucose sensor. The invention includes the polymer composition, sensor systems formed from this polymer...

Development of an enzyme free glucose sensor based on copper oxide-graphene composite by using green reducing agent ascorbic acid

Science.gov (United States)

Palve, Yogesh Pandit; Jha, Neetu

2018-05-01

In this research work we have developed high sensitive and selective glucose sensor based on copper oxide-graphene composite which is prepared by green synthesis method and used for nonenzymatic glucose sensor. In present paper we report that present method highly selective, simple, efficient, accurate, ecofriendly, less toxic. The prepared composite were characterized by material characterization like SEM, XRD and also by electrochemical characterization like CV, chronoamperometry represents that copper oxide-graphene shows excellent electrocatalytic activity towards glucose, exhibiting a good sensitivity of 103.84 µA mM-1 cm-2, a fast response time 2s, a low detection limit 0.00033µM and linear range from 10 µM-3000 µM. The present sensor can successfully apply for determination of glucose concentration in human blood sample.

A CuNi/C Nanosheet Array Based on a Metal-Organic Framework Derivate as a Supersensitive Non-Enzymatic Glucose Sensor

Science.gov (United States)

Zhang, Li; Ye, Chen; Li, Xu; Ding, Yaru; Liang, Hongbo; Zhao, Guangyu; Wang, Yan

2018-06-01

Bimetal catalysts are good alternatives for non-enzymatic glucose sensors owing to their low cost, high activity, good conductivity, and ease of fabrication. In the present study, a self-supported CuNi/C electrode prepared by electrodepositing Cu nanoparticles on a Ni-based metal-organic framework (MOF) derivate was used as a non-enzymatic glucose sensor. The porous construction and carbon scaffold inherited from the Ni-MOF guarantee good kinetics of the electrode process in electrochemical glucose detection. Furthermore, Cu nanoparticles disturb the array structure of MOF derived films and evidently enhance their electrochemical performances in glucose detection. Electrochemical measurements indicate that the CuNi/C electrode possesses a high sensitivity of 17.12 mA mM-1 cm-2, a low detection limit of 66.67 nM, and a wider linearity range from 0.20 to 2.72 mM. Additionally, the electrode exhibits good reusability, reproducibility, and stability, thereby catering to the practical use of glucose sensors. Similar values of glucose concentrations in human blood serum samples are detected with our electrode and with the method involving glucose-6-phosphate dehydrogenase; the results further demonstrate the practical feasibility of our electrode.

Glucose Sensor MdHXK1 Phosphorylates and Stabilizes MdbHLH3 to Promote Anthocyanin Biosynthesis in Apple

Science.gov (United States)

Hu, Da-Gang; Zhang, Quan-Yan; An, Jian-Ping; You, Chun-Xiang; Hao, Yu-Jin

2016-01-01

Glucose induces anthocyanin accumulation in many plant species; however, the molecular mechanism involved in this process remains largely unknown. Here, we found that apple hexokinase MdHXK1, a glucose sensor, was involved in sensing exogenous glucose and regulating anthocyanin biosynthesis. In vitro and in vivo assays suggested that MdHXK1 interacted directly with and phosphorylated an anthocyanin-associated bHLH transcription factor (TF) MdbHLH3 at its Ser361 site in response to glucose. Furthermore, both the hexokinase_2 domain and signal peptide are crucial for the MdHXK1-mediated phosphorylation of MdbHLH3. Moreover, phosphorylation modification stabilized MdbHLH3 protein and enhanced its transcription of the anthocyanin biosynthesis genes, thereby increasing anthocyanin biosynthesis. Finally, a series of transgenic analyses in apple calli and fruits demonstrated that MdHXK1 controlled glucose-induced anthocyanin accumulation at least partially, if not completely, via regulating MdbHLH3. Overall, our findings provide new insights into the mechanism of the glucose sensor HXK1 modulation of anthocyanin accumulation, which occur by directly regulating the anthocyanin-related bHLH TFs in response to a glucose signal in plants. PMID:27560976

Glucose Sensor MdHXK1 Phosphorylates and Stabilizes MdbHLH3 to Promote Anthocyanin Biosynthesis in Apple.

Directory of Open Access Journals (Sweden)

Da-Gang Hu

2016-08-01

Full Text Available Glucose induces anthocyanin accumulation in many plant species; however, the molecular mechanism involved in this process remains largely unknown. Here, we found that apple hexokinase MdHXK1, a glucose sensor, was involved in sensing exogenous glucose and regulating anthocyanin biosynthesis. In vitro and in vivo assays suggested that MdHXK1 interacted directly with and phosphorylated an anthocyanin-associated bHLH transcription factor (TF MdbHLH3 at its Ser361 site in response to glucose. Furthermore, both the hexokinase_2 domain and signal peptide are crucial for the MdHXK1-mediated phosphorylation of MdbHLH3. Moreover, phosphorylation modification stabilized MdbHLH3 protein and enhanced its transcription of the anthocyanin biosynthesis genes, thereby increasing anthocyanin biosynthesis. Finally, a series of transgenic analyses in apple calli and fruits demonstrated that MdHXK1 controlled glucose-induced anthocyanin accumulation at least partially, if not completely, via regulating MdbHLH3. Overall, our findings provide new insights into the mechanism of the glucose sensor HXK1 modulation of anthocyanin accumulation, which occur by directly regulating the anthocyanin-related bHLH TFs in response to a glucose signal in plants.

Dual functional rhodium oxide nanocorals enabled sensor for both non-enzymatic glucose and solid-state pH sensing.

Science.gov (United States)

Dong, Qiuchen; Huang, Yikun; Song, Donghui; Wu, Huixiang; Cao, Fei; Lei, Yu

2018-07-30

Both pH-sensitive and glucose-responsive rhodium oxide nanocorals (Rh 2 O 3 NCs) were synthesized through electrospinning followed by high-temperature calcination. The as-prepared Rh 2 O 3 NCs were systematically characterized using various advanced techniques including scanning electron microscopy, X-ray powder diffraction and Raman spectroscopy, and then employed as a dual functional nanomaterial to fabricate a dual sensor for both non-enzymatic glucose sensing and solid-state pH monitoring. The sensing performance of the Rh 2 O 3 NCs based dual sensor toward pH and glucose was evaluated using open circuit potential, cyclic voltammetry and amperometric techniques, respectively. The results show that the as-prepared Rh 2 O 3 NCs not only maintain accurate and reversible pH sensitivity of Rh 2 O 3 , but also demonstrate a good electrocatalytic activity toward glucose oxidation in alkaline medium with a sensitivity of 11.46 μA mM -1 cm -2 , a limit of detection of 3.1 μM (S/N = 3), and a reasonable selectivity against various interferents in non-enzymatic glucose detection. Its accuracy in determining glucose in human serum samples was further demonstrated. These features indicate that the as-prepared Rh 2 O 3 NCs hold great promise as a dual-functional sensing material in the development of a high-performance sensor forManjakkal both solid-state pH and non-enzymatic glucose sensing. Copyright © 2018 Elsevier B.V. All rights reserved.

A high performance non-enzymatic glucose sensor based on nickel hydroxide modified nitrogen-incorporated nanodiamonds.

Science.gov (United States)

Ko, Chih-Yu; Huang, Jin-Hua; Raina, Supil; Kang, Weng P

2013-06-07

A highly selective, sensitive, and stable non-enzymatic glucose sensor based on Ni hydroxide modified nitrogen-incorporated nanodiamonds (Ni(OH)2-NND) was developed. The sensor was fabricated by e-beam evaporation of a thin Ni film on NND followed by the growth of Ni(OH)2 using an electrochemical process. It was found that the Ni film thickness greatly affects the morphology and electro-catalytic activity of the as-synthesized electrode for non-enzymatic glucose oxidation. Owing to its nanostructure characteristics, the best sensor fabricated by 150 nm Ni deposition showed two wide response ranges, namely, 0.02-1 mM and 1-9 mM, with sensitivities of 3.20 and 1.41 mA mM(-1) cm(-2), respectively, and a detection limit of 1.2 μM (S/N = 3). The sensor also showed good long-term stability as well as high selectivity in the presence of interferences such as ascorbic acid, acetaminophen, and uric acid. This finding reveals the possibility of exploiting the NND as an electrochemical biosensor platform where high performance addressable sensor arrays could be built.

The glucose enzyme electrode: is simple peroxide detection at a needle sensor acceptable?

Science.gov (United States)

Vadgama, P; Mullen, W; Churchouse, S; Battersby, C

1988-01-01

Needle type devices have been fabricated based on Pt anodes (to detect H2O2) mounted with stainless steel needles to act as the reference. Coating of these devices with glucose oxidase allowed glucose measurement, but with high dependence on stirring and background pO2 levels as well as a restricted glucose assay range. By wet dipcoating of microporous polyurethane or the application of preformed porous membranes, the linear range has been extended up to 70 mM glucose, to give minimal pO2 dependence and insensitivity to stirring. With incorporation of polyethersulphone membranes, blood measurement was possible with high selectivity (y = 0.954x + 0.202, r = 0.991, n = 48). This establishes that simple peroxide detection at a needle sensor is acceptable, and can now be focussed on increasing biocompatibility.

O-GlcNAc transferase regulates transcriptional activity of human Oct4.

Science.gov (United States)

Constable, Sandii; Lim, Jae-Min; Vaidyanathan, Krithika; Wells, Lance

2017-10-01

O-linked β-N-acetylglucosamine (O-GlcNAc) is a single sugar modification found on many different classes of nuclear and cytoplasmic proteins. Addition of this modification, by the enzyme O-linked N-acetylglucosamine transferase (OGT), is dynamic and inducible. One major class of proteins modified by O-GlcNAc is transcription factors. O-GlcNAc regulates transcription factor properties through a variety of different mechanisms including localization, stability and transcriptional activation. Maintenance of embryonic stem (ES) cell pluripotency requires tight regulation of several key transcription factors, many of which are modified by O-GlcNAc. Octamer-binding protein 4 (Oct4) is one of the key transcription factors required for pluripotency of ES cells and more recently, the generation of induced pluripotent stem (iPS) cells. The action of Oct4 is modulated by the addition of several post-translational modifications, including O-GlcNAc. Previous studies in mice found a single site of O-GlcNAc addition responsible for transcriptional regulation. This study was designed to determine if this mechanism is conserved in humans. We mapped 10 novel sites of O-GlcNAc attachment on human Oct4, and confirmed a role for OGT in transcriptional activation of Oct4 at a site distinct from that found in mouse that allows distinction between different Oct4 target promoters. Additionally, we uncovered a potential new role for OGT that does not include its catalytic function. These results confirm that human Oct4 activity is being regulated by OGT by a mechanism that is distinct from mouse Oct4. © The Author 2017. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Fabrication of a Microneedle/CNT Hierarchical Micro/Nano Surface Electrochemical Sensor and Its In-Vitro Glucose Sensing Characterization

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Youngsam Yoon

2013-12-01

Full Text Available We report fabrication of a microneedle-based three-electrode integrated electrochemical sensor and in-vitro characterization of this sensor for glucose sensing applications. A piece of silicon was sequentially dry and wet etched to form a 15 × 15 array of tall (approximately 380 µm sharp silicon microneedles. Iron catalyst was deposited through a SU-8 shadow mask to form the working electrode and counter electrode. A multi-walled carbon nanotube forest was grown directly on the silicon microneedle array and platinum nano-particles were electrodeposited. Silver was deposited on the Si microneedle array through another shadow mask and chlorinated to form a Ag/AgCl reference electrode. The 3-electrode electrochemical sensor was tested for various glucose concentrations in the range of 3~20 mM in 0.01 M phosphate buffered saline (PBS solution. The sensor’s amperometric response to the glucose concentration is linear and its sensitivity was found to be 17.73 ± 3 μA/mM-cm2. This microneedle-based sensor has a potential to be used for painless diabetes testing applications.

Label-free glucose detection using cantilever sensor technology based on gravimetric detection principles.

Science.gov (United States)

Hsieh, Shuchen; Hsieh, Shu-Ling; Hsieh, Chiung-Wen; Lin, Po-Chiao; Wu, Chun-Hsin

2013-01-01

Efficient maintenance of glucose homeostasis is a major challenge in diabetes therapy, where accurate and reliable glucose level detection is required. Though several methods are currently used, these suffer from impaired response and often unpredictable drift, making them unsuitable for long-term therapeutic practice. In this study, we demonstrate a method that uses a functionalized atomic force microscope (AFM) cantilever as the sensor for reliable glucose detection with sufficient sensitivity and selectivity for clinical use. We first modified the AFM tip with aminopropylsilatrane (APS) and then adsorbed glucose-specific lectin concanavalin A (Con A) onto the surface. The Con A/APS-modified probes were then used to detect glucose by monitoring shifts in the cantilever resonance frequency. To confirm the molecule-specific interaction, AFM topographical images were acquired of identically treated silicon substrates which indicated a specific attachment for glucose-Con A and not for galactose-Con A. These results demonstrate that by monitoring the frequency shift of the AFM cantilever, this sensing system can detect the interaction between Con A and glucose, one of the biomolecule recognition processes, and may assist in the detection and mass quantification of glucose for clinical applications with very high sensitivity.

Glucose Sensing

CERN Document Server

Geddes, Chris D

2006-01-01

Topics in Fluorescence Spectroscopy, Glucose Sensing is the eleventh volume in the popular series Topics in Fluorescence Spectroscopy, edited by Drs. Chris D. Geddes and Joseph R. Lakowicz. This volume incorporates authoritative analytical fluorescence-based glucose sensing reviews specialized enough to be attractive to professional researchers, yet also appealing to the wider audience of scientists in related disciplines of fluorescence. Glucose Sensing is an essential reference for any lab working in the analytical fluorescence glucose sensing field. All academics, bench scientists, and industry professionals wishing to take advantage of the latest and greatest in the continuously emerging field of glucose sensing, and diabetes care & management, will find this volume an invaluable resource. Topics in Fluorescence Spectroscopy Volume 11, Glucose Sensing Chapters include: Implantable Sensors for Interstitial Fluid Smart Tattoo Glucose Sensors Optical Enzyme-based Glucose Biosensors Plasmonic Glucose Sens...

Thermo-induced modifications and selective accumulation of glucose-conjugated magnetic nanoparticles in vivo in rats - increasing the effectiveness of magnetic-assisted therapy - pilot study.

Science.gov (United States)

Traikov, L; Antonov, I; Gerou, A; Vesselinova, L; Hadjiolova, R; Raynov, J

2015-09-01

Ferro-Magnetic nanoparticles (Fe-MNP) have gained a lot of attention in biomedical and industrial applications due to their biocompatibility, ease of surface modification and paramagnetic properties. The basic idea of our study is whether it is possible to use glucose-conjugate Fe-MNP (Glc-Fe-MNP) for targeting and more accurate focusing in order to increase the effect of high-frequency electromagnetic fields induced hyperthermia in solid tumors. Tumors demonstrate high metabolic activity for glucose in comparison with other somatic cells.Increasing of accumulation of glucose conjugated (Glc)-Fe-MNP on tumor site and precision of radio frequency electro-magnetic field (RF-EMF) energy absorption in solid tumors, precede RF-EMF induced hyperthermia. Rat model for monitoring the early development of breast cancer. Twenty female Wistar rats (MU-line-6171) were divided into two groups of 10 rats that were either treated with N-methyl-N-nitrosourea to induce breast cancer and 10 with carrageenan to induce inflammation (control). Glc-Fe-MNP can offer a solution to increase hyperthermia effect to the desired areas in the body by accumulation and increasing local concentration due to high tissue metabolic assimilation. In this condition, it is considered that the magnetization of the nanoparticles is a single-giant magnetic moment, the sum of all the individual magnetic moments and is proportional to the concentration of Glc-Fe-MNP.

Brain glucose transport and phosphorylation under acute insulin-induced hypoglycemia in mice: an 18F-FDG PET study.

Science.gov (United States)

Alf, Malte F; Duarte, João M N; Schibli, Roger; Gruetter, Rolf; Krämer, Stefanie D

2013-12-01

We addressed the questions of how cerebral glucose transport and phosphorylation change under acute hypoglycemia and what the underlying mechanisms of adaptation are. Quantitative (18)F-FDG PET combined with the acquisition of real-time arterial input function was performed on mice. Hypoglycemia was induced and maintained by insulin infusion. PET data were analyzed with the 2-tissue-compartment model for (18)F-FDG, and the results were evaluated with Michaelis-Menten saturation kinetics. Glucose clearance from plasma to brain (K1,glc) and the phosphorylation rate constant increased with decreasing plasma glucose (Gp), in particular at a Gp of less than 2.5 mmol/L. Estimated cerebral glucose extraction ratios taking into account an increased cerebral blood flow (CBF) at a Gp of less than 2 mmol/L were between 0.14 and 0.79. CBF-normalized K1,glc values were in agreement with saturation kinetics. Phosphorylation rate constants indicated intracellular glucose depletion at a Gp of less than 2-3 mmol/L. When brain regions were compared, glucose transport under hypoglycemia was lowest in the hypothalamus. Alterations in glucose transport and phosphorylation, as well as intracellular glucose depletion, under acute hypoglycemia can be modeled by saturation kinetics taking into account an increase in CBF. Distinct transport kinetics in the hypothalamus may be involved in its glucose-sensing function.

Dynamic O-GlcNAc cycling at promoters of Caenorhabditis elegans genes regulating longevity, stress, and immunity.

Science.gov (United States)

Love, Dona C; Ghosh, Salil; Mondoux, Michelle A; Fukushige, Tetsunari; Wang, Peng; Wilson, Mark A; Iser, Wendy B; Wolkow, Catherine A; Krause, Michael W; Hanover, John A

2010-04-20

Nutrient-driven O-GlcNAcylation of key components of the transcription machinery may epigenetically modulate gene expression in metazoans. The global effects of GlcNAcylation on transcription can be addressed directly in C. elegans because knockouts of the O-GlcNAc cycling enzymes are viable and fertile. Using anti-O-GlcNAc ChIP-on-chip whole-genome tiling arrays on wild-type and mutant strains, we detected over 800 promoters where O-GlcNAc cycling occurs, including microRNA loci and multigene operons. Intriguingly, O-GlcNAc-marked promoters are biased toward genes associated with PIP3 signaling, hexosamine biosynthesis, and lipid/carbohydrate metabolism. These marked genes are linked to insulin-like signaling, metabolism, aging, stress, and pathogen-response pathways in C. elegans. Whole-genome transcriptional profiling of the O-GlcNAc cycling mutants confirmed dramatic deregulation of genes in these key pathways. As predicted, the O-GlcNAc cycling mutants show altered lifespan and UV stress susceptibility phenotypes. We propose that O-GlcNAc cycling at promoters participates in a molecular program impacting nutrient-responsive pathways in C. elegans, including stress, pathogen response, and adult lifespan. The observed impact of O-GlcNAc cycling on both signaling and transcription in C. elegans has important implications for human diseases of aging, including diabetes and neurodegeneration.

Evaluation and Computational Characterization of the Faciliated Transport of Glc Carbon C-1 Oxime Reactivators Across a Blood Brain Barrier Model

Science.gov (United States)

2013-01-01

blood brain barrier (BBB) to reactivate inhibited brain acetylcholinesterase (AChE). We selected glucose (Glc) transporters (GLUT) for this purpose as...Eur. J. Pharm. 332 (1997) 43–52. [4] N.J. Abbott , L. Ronnback, E. Hansson, Astrocyte-endothelial interactions at the blood –brain barrier, Nat. Rev...5a. CONTRACT NUMBER oxime reactivators across a blood brain barrier model 5b. GRANT NUMBER 1.E005.08.WR 5c. PROGRAM ELEMENT NUMBER 6. AUTHOR(S

2-Deoxyglucose induces the expression of thioredoxin interacting protein (TXNIP) by increasing O-GlcNAcylation – Implications for targeting the Warburg effect in cancer cells

Energy Technology Data Exchange (ETDEWEB)

Hong, Shin Yee; Hagen, Thilo, E-mail: bchth@nus.edu.sg

2015-10-02

The high proliferation rate of cancer cells and the microenvironment in the tumor tissue require the reprogramming of tumor cell metabolism. The major mechanism of metabolic reprogramming in cancer cells is the Warburg effect, defined as the preferential utilization of glucose via glycolysis even in the presence of oxygen. Targeting the Warburg effect is considered as a promising therapeutic strategy in cancer therapy. In this regard, the glycolytic inhibitor 2-deoxyglucose (2DG) has been evaluated clinically. 2DG exerts its effect by directly inhibiting glycolysis at the level of hexokinase and phosphoglucoisomerase. In addition, 2DG is also known to induce the expression of thioredoxin interacting protein (TXNIP), a tumor suppressor protein and an important negative regulator of cellular glucose uptake. Hence, characterization of the mechanism through which 2DG regulates TXNIP expression may reveal novel approaches to target the Warburg effect in cancer cells. Therefore, in this study we sought to test various hypotheses for the mechanistic basis of the 2DG dependent TXNIP regulation. We have shown that 2DG induced TXNIP expression is independent of carbohydrate response element mediated transcription. Furthermore, the induction of TXNIP is neither dependent on the ability of 2DG to deplete cellular ATP nor to cause endoplasmic reticulum stress. We found that the 2DG induced TXNIP expression is at least in part dependent on the inhibition of the O-GlcNAcase enzyme and the accumulation of O-GlcNAc modified proteins. These results have implications for the identification of therapeutic targets to increase TXNIP expression in cancer. - Highlights: • 2DG increases TXNIP expression at the mRNA and protein level. • The effect of 2DG on TXNIP is independent of ChoRE mediated transcription. • 2DG induces TXNIP independent of ER stress induction and ATP depletion. • 2DG inhibits OGA and leads to accumulation of O-GlcNAcylated proteins. • The upregulation of

O-GlcNAcylation modulates PKA-CREB signaling in a manner specific to PKA catalytic subunit isoforms.

Science.gov (United States)

Jin, Nana; Ma, Denglei; Gu, Jianlan; Shi, Jianhua; Xu, Xiaotao; Iqbal, Khalid; Gong, Cheng-Xin; Liu, Fei; Chu, Dandan

2018-02-26

O-GlcNAcylation is a post-translational modification of proteins. Protein kinase A (PKA)-cAMP response element binding protein (CREB) signaling plays critical roles in multiple biological processes. Isoforms α and β of PKA catalytic subunit (PKAc) and CREB are modified by O-GlcNAcylation. In the present study, we determined the role of O-GlcNAcylation in PKAc isoform-specific CREB signaling. We found that up-regulation of O-GlcNAcylation enhanced CREB phosphorylation, but suppressed CREB expression in exogenous PKAc isoform-unspecific manner. PKAc isoforms affected exogenous expression of OGT or OGA and protein O-GlcNAcylation differently. Up-regulation of O-GlcNAcylation did not significantly affect net PKAcα-CREB signaling, but enhanced PKAcβ-CREB signaling. The role of O-GlcNAcylation in PKA-CREB signaling was desensitized by insulin treatment. This study suggests a role of O-GlcNAcylation in PKA-CREB signaling by affecting phosphorylation of CREB in a PKAc isoform-specific manner. Copyright © 2018 Elsevier Inc. All rights reserved.

Neuronal calcium sensor synaptotagmin-9 is not involved in the regulation of glucose homeostasis or insulin secretion.

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Natalia Gustavsson

Full Text Available BACKGROUND: Insulin secretion is a complex and highly regulated process. It is well established that cytoplasmic calcium is a key regulator of insulin secretion, but how elevated intracellular calcium triggers insulin granule exocytosis remains unclear, and we have only begun to define the identities of proteins that are responsible for sensing calcium changes and for transmitting the calcium signal to release machineries. Synaptotagmins are primarily expressed in brain and endocrine cells and exhibit diverse calcium binding properties. Synaptotagmin-1, -2 and -9 are calcium sensors for fast neurotransmitter release in respective brain regions, while synaptotagmin-7 is a positive regulator of calcium-dependent insulin release. Unlike the three neuronal calcium sensors, whose deletion abolished fast neurotransmitter release, synaptotagmin-7 deletion resulted in only partial loss of calcium-dependent insulin secretion, thus suggesting that other calcium-sensors must participate in the regulation of insulin secretion. Of the other synaptotagmin isoforms that are present in pancreatic islets, the neuronal calcium sensor synaptotagmin-9 is expressed at the highest level after synaptotagmin-7. METHODOLOGY/PRINCIPAL FINDINGS: In this study we tested whether synaptotagmin-9 participates in the regulation of glucose-stimulated insulin release by using pancreas-specific synaptotagmin-9 knockout (p-S9X mice. Deletion of synaptotagmin-9 in the pancreas resulted in no changes in glucose homeostasis or body weight. Glucose tolerance, and insulin secretion in vivo and from isolated islets were not affected in the p-S9X mice. Single-cell capacitance measurements showed no difference in insulin granule exocytosis between p-S9X and control mice. CONCLUSIONS: Thus, synaptotagmin-9, although a major calcium sensor in the brain, is not involved in the regulation of glucose-stimulated insulin release from pancreatic β-cells.

Neuronal calcium sensor synaptotagmin-9 is not involved in the regulation of glucose homeostasis or insulin secretion.

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Gustavsson, Natalia; Wang, Xiaorui; Wang, Yue; Seah, Tingting; Xu, Jun; Radda, George K; Südhof, Thomas C; Han, Weiping

2010-11-09

Insulin secretion is a complex and highly regulated process. It is well established that cytoplasmic calcium is a key regulator of insulin secretion, but how elevated intracellular calcium triggers insulin granule exocytosis remains unclear, and we have only begun to define the identities of proteins that are responsible for sensing calcium changes and for transmitting the calcium signal to release machineries. Synaptotagmins are primarily expressed in brain and endocrine cells and exhibit diverse calcium binding properties. Synaptotagmin-1, -2 and -9 are calcium sensors for fast neurotransmitter release in respective brain regions, while synaptotagmin-7 is a positive regulator of calcium-dependent insulin release. Unlike the three neuronal calcium sensors, whose deletion abolished fast neurotransmitter release, synaptotagmin-7 deletion resulted in only partial loss of calcium-dependent insulin secretion, thus suggesting that other calcium-sensors must participate in the regulation of insulin secretion. Of the other synaptotagmin isoforms that are present in pancreatic islets, the neuronal calcium sensor synaptotagmin-9 is expressed at the highest level after synaptotagmin-7. In this study we tested whether synaptotagmin-9 participates in the regulation of glucose-stimulated insulin release by using pancreas-specific synaptotagmin-9 knockout (p-S9X) mice. Deletion of synaptotagmin-9 in the pancreas resulted in no changes in glucose homeostasis or body weight. Glucose tolerance, and insulin secretion in vivo and from isolated islets were not affected in the p-S9X mice. Single-cell capacitance measurements showed no difference in insulin granule exocytosis between p-S9X and control mice. Thus, synaptotagmin-9, although a major calcium sensor in the brain, is not involved in the regulation of glucose-stimulated insulin release from pancreatic β-cells.

A high-performance liquid chromatography-based radiometric assay for sucrose-phosphate synthase and other UDP-glucose requiring enzymes

International Nuclear Information System (INIS)

Salvucci, M.E.; Crafts-Brandner, S.J.

1991-01-01

A method for product analysis that eliminates a problematic step in the radiometric sucrose-phosphate synthase assay is described. The method uses chromatography on a boronate-derivatized high-performance liquid chromatography column to separate the labeled product, [14C]sucrose phosphate, from unreacted uridine 5'-diphosphate-[14C]glucose (UDP-Glc). Direct separation of these compounds eliminates the need for treatment of the reaction mixtures with alkaline phosphatase, thereby avoiding the problem of high background caused by contaminating phosphodiesterase activity in alkaline phosphatase preparations. The method presented in this paper can be applied to many UDP-Glc requiring enzymes; here the authors show its use for determining the activities of sucrose-phosphate synthase, sucrose synthase, and uridine diphosphate-glucose pyrophosphorylase in plant extracts

A fine pointed glucose oxidase immobilized electrode for low-invasive amperometric glucose monitoring.

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Li, Jiang; Koinkar, Pankaj; Fuchiwaki, Yusuke; Yasuzawa, Mikito

2016-12-15

A low invasive type glucose sensor, which has a sensing region at the tip of a fine pointed electrode, was developed for continuous glucose monitoring. Platinum-iridium alloy electrode with a surface area of 0.045mm(2) was settled at the middle of pointed PEEK (Polyetheretherketone) tubing and was employed as sensing electrode. Electrodeposition of glucose oxidase in the presence of surfactant, Triton X-100, was performed for high-density enzyme immobilization followed by the electropolymerization of o-phenylenediamine for the formation of functional entrapping and permselective polymer membrane. Ag/AgCl film was coated on the surface of PEEK tubing as reference electrode. Amperometric responses of the prepared sensors to glucose were measured at a potential of 0.60V (vs. Ag/AgCl). The prepared electrode showed the sensitivity of 2.55μA/cm(2) mM with high linearity of 0.9986, within the glucose concentration range up to 21mM. The detection limit (S/N=3) was determined to be 0.11mM. The glucose sensor properties were evaluated in phosphate buffer solution and in vivo monitoring by the implantation of the sensors in rabbit, while conventional needle type sensors as a reference were used. The results showed that change in output current of the proposed sensor fluctuated similar with one in output current of the conventional needle type sensors, which was also in similar accordance with actual blood sugar level measured by commercially glucose meter. One-point calibration method was used to calibrate the sensor output current. Copyright © 2016 Elsevier B.V. All rights reserved.

Measurement of Non-Invasive Blood Glucose Level Based Sensor Color TCS3200 and Arduino

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Kurniadi Wardana, Humaidillah; Indahwati, Elly; Arifah Fitriyah, Lina

2018-04-01

Design and measurement of Arduino-based urinary (non-invasive) urine glucose using RGB tcs3200 sensor. This research was conducted by making use of the urine in diabetes patients detected by sensor colours then measured levels of colour based on the RGB colour of the urine of diabetics. The detection is done on 4 urine samples with each consisting of 3 diabetics and 1 non-diabetics. Equipment used in this research, among others, Arduino Uno, colour sensor tcs3200, LCD 16x4. The results showed that the detection of RGB values in diabetics 230 with blue and not diabetics 200 with red.

The role of O-GlcNAc signaling in the pathogenesis of diabetic retinopathy.

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Semba, Richard D; Huang, Hu; Lutty, Gerard A; Van Eyk, Jennifer E; Hart, Gerald W

2014-04-01

Diabetic retinopathy is a leading cause of blindness worldwide. Despite laser and surgical treatments, antiangiogenic and other therapies, and strict metabolic control, many patients progress to visual impairment and blindness. New insights are needed into the pathophysiology of diabetic retinopathy in order to develop new methods to improve the detection and treatment of disease and the prevention of blindness. Hyperglycemia and diabetes result in increased flux through the hexosamine biosynthetic pathway, which, in turn, results in increased PTM of Ser/Thr residues of proteins by O-linked β-N-acetylglucosamine (O-GlcNAc). O-GlcNAcylation is involved in regulation of many nuclear and cytoplasmic proteins in a manner similar to protein phosphorylation. Altered O-GlcNAc signaling has been implicated in the pathogenesis of diabetes and may play an important role in the pathogenesis of diabetic retinopathy. The goal of this review is to summarize the biology of the hexosamine biosynthesis pathway and O-GlcNAc signaling, to present the current evidence for the role of O-GlcNAc signaling in diabetes and diabetic retinopathy, and to discuss future directions for research on O-GlcNAc in the pathogenesis of diabetic retinopathy. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

The Role of PTP1B O-GlcNAcylation in Hepatic Insulin Resistance

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Yun Zhao

2015-09-01

Full Text Available Protein tyrosine phosphatase 1B (PTP1B, which can directly dephosphorylate both the insulin receptor and insulin receptor substrate 1 (IRS-1, thereby terminating insulin signaling, reportedly plays an important role in insulin resistance. Accumulating evidence has demonstrated that O-GlcNAc modification regulates functions of several important components of insulin signal pathway. In this study, we identified that PTP1B is modified by O-GlcNAcylation at three O-GlcNAc sites (Ser104, Ser201, and Ser386. Palmitate acid (PA impaired the insulin signaling, indicated by decreased phosphorylation of both serine/threonine-protein kinase B (Akt and glycogen synthase kinase 3 beta (GSK3β following insulin administration, and upregulated PTP1B O-GlcNAcylation in HepG2 cells. Compared with the wild-type, intervention PTP1B O-GlcNAcylation by site-directed gene mutation inhibited PTP1B phosphatase activity, resulted in a higher level of phosphorylated Akt and GSK3β, recovered insulin sensitivity, and improved lipid deposition in HepG2 cells. Taken together, our research showed that O-GlcNAcylation of PTP1B can influence insulin signal transduction by modulating its own phosphatase activity, which participates in the process of hepatic insulin resistance.

XBP1 (X-Box-Binding Protein-1)-Dependent O-GlcNAcylation Is Neuroprotective in Ischemic Stroke in Young Mice and Its Impairment in Aged Mice Is Rescued by Thiamet-G.

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Jiang, Meng; Yu, Shu; Yu, Zhui; Sheng, Huaxin; Li, Ying; Liu, Shuai; Warner, David S; Paschen, Wulf; Yang, Wei

2017-06-01

Impaired protein homeostasis induced by endoplasmic reticulum dysfunction is a key feature of a variety of age-related brain diseases including stroke. To restore endoplasmic reticulum function impaired by stress, the unfolded protein response is activated. A key unfolded protein response prosurvival pathway is controlled by the endoplasmic reticulum stress sensor (inositol-requiring enzyme-1), XBP1 (downstream X-box-binding protein-1), and O-GlcNAc (O-linked β-N-acetylglucosamine) modification of proteins (O-GlcNAcylation). Stroke impairs endoplasmic reticulum function, which activates unfolded protein response. The rationale of this study was to explore the potentials of the IRE1/XBP1/O-GlcNAc axis as a target for neuroprotection in ischemic stroke. Mice with Xbp1 loss and gain of function in neurons were generated. Stroke was induced by transient or permanent occlusion of the middle cerebral artery in young and aged mice. Thiamet-G was used to increase O-GlcNAcylation. Deletion of Xbp1 worsened outcome after transient and permanent middle cerebral artery occlusion. After stroke, O-GlcNAcylation was activated in neurons of the stroke penumbra in young mice, which was largely Xbp1 dependent. This activation of O-GlcNAcylation was impaired in aged mice. Pharmacological increase of O-GlcNAcylation before or after stroke improved outcome in both young and aged mice. Our study indicates a critical role for the IRE1/XBP1 unfolded protein response branch in stroke outcome. O-GlcNAcylation is a prosurvival pathway that is activated in the stroke penumbra in young mice but impaired in aged mice. Boosting prosurvival pathways to counterbalance the age-related decline in the brain's self-healing capacity could be a promising strategy to improve ischemic stroke outcome in aged brains. © 2017 American Heart Association, Inc.

Mutations in N-acetylglucosamine (O-GlcNAc) transferase in patients with X-linked intellectual disability

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Willems, A.P.; Gundogdu, M.; Kempers, M.J.E.; Giltay, J.C.; Pfundt, R.P.; Elferink, M.; Loza, B.F.; Fuijkschot, J.; Ferenbach, A.T.; Gassen, K.L. van; Aalten, D.M.F. van; Lefeber, D.J.

2017-01-01

N-Acetylglucosamine (O-GlcNAc) transferase (OGT) regulates protein O-GlcNAcylation, an essential and dynamic post-translational modification. The O-GlcNAc modification is present on numerous nuclear and cytosolic proteins and has been implicated in essential cellular functions such as signaling and

Sweet taste signaling functions as a hypothalamic glucose sensor

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Xueying Ren

2009-06-01

Full Text Available Brain glucosensing is essential for normal body glucose homeostasis and neuronal function. However, the exact signaling mechanisms involved in the neuronal sensing of extracellular glucose levels remain poorly understood. Of particular interest is the identification of candidate membrane molecular sensors allowing neurons to change firing rates independently of intracellular glucose metabolism. Here we describe for the first time the expression of the taste receptor genes Tas1r1, Tas1r2 and Tas1r3, and their associated G-protein genes, in the mammalian brain. Neuronal expression of taste genes was detected in different nutrient-sensing forebrain regions, including the paraventricular and arcuate nuclei of the hypothalamus, the CA fields and dentate gyrus of the hippocampus, the habenula, and cortex. Expression was also observed in the intra-ventricular epithelial cells of the choroid plexus. These same regions were found to express the corresponding gene products that form the heterodimeric T1R2/T1R3 and T1R1/T1R3 sweet and L-amino acid taste G-protein coupled receptors, respectively. These regions were also found to express the taste G-protein α-Gustducin. Moreover, in vivo studies in mice demonstrate that the hypothalamic expression of taste-related genes is regulated by the nutritional state of the animal, with food deprivation significantly increasing expression levels of Tas1r1 and Tas1r2 in hypothalamus, but not in cortex. Furthermore, exposing mouse hypothalamic cells to a low-glucose medium, while maintaining normal L-amino acid concentrations, specifically resulted in higher expression levels of the sweet-associated gene Tas1r2. This latter effect was reversed by adding the non-metabolizable artificial sweetener sucralose to the low-glucose medium, indicating that taste-like signaling in hypothalamic neurons does not require intracellular glucose oxidation. Our findings suggest that the G-protein coupled sweet receptor T1R2/T1R3 is a

An OGA-Resistant Probe Allows Specific Visualization and Accurate Identification of O-GlcNAc-Modified Proteins in Cells.

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Li, Jing; Wang, Jiajia; Wen, Liuqing; Zhu, He; Li, Shanshan; Huang, Kenneth; Jiang, Kuan; Li, Xu; Ma, Cheng; Qu, Jingyao; Parameswaran, Aishwarya; Song, Jing; Zhao, Wei; Wang, Peng George

2016-11-18

O-linked β-N-acetyl-glucosamine (O-GlcNAc) is an essential and ubiquitous post-translational modification present in nucleic and cytoplasmic proteins of multicellular eukaryotes. The metabolic chemical probes such as GlcNAc or GalNAc analogues bearing ketone or azide handles, in conjunction with bioorthogonal reactions, provide a powerful approach for detecting and identifying this modification. However, these chemical probes either enter multiple glycosylation pathways or have low labeling efficiency. Therefore, selective and potent probes are needed to assess this modification. We report here the development of a novel probe, 1,3,6-tri-O-acetyl-2-azidoacetamido-2,4-dideoxy-d-glucopyranose (Ac 3 4dGlcNAz), that can be processed by the GalNAc salvage pathway and transferred by O-GlcNAc transferase (OGT) to O-GlcNAc proteins. Due to the absence of a hydroxyl group at C4, this probe is less incorporated into α/β 4-GlcNAc or GalNAc containing glycoconjugates. Furthermore, the O-4dGlcNAz modification was resistant to the hydrolysis of O-GlcNAcase (OGA), which greatly enhanced the efficiency of incorporation for O-GlcNAcylation. Combined with a click reaction, Ac 3 4dGlcNAz allowed the selective visualization of O-GlcNAc in cells and accurate identification of O-GlcNAc-modified proteins with LC-MS/MS. This probe represents a more potent and selective tool in tracking, capturing, and identifying O-GlcNAc-modified proteins in cells and cell lysates.

Enzymatic glucose sensor based on Au nanoparticle and plant-like ZnO film modified electrode

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Tian, Kun [Nanostructured Materials Research Laboratory, Department of Materials Science and Engineering, University of Utah, Salt Lake City, UT 84112 (United States); Alex, Saji [Nanostructured Materials Research Laboratory, Department of Materials Science and Engineering, University of Utah, Salt Lake City, UT 84112 (United States); Department of Chemistry, Government College for Women, Thiruvananthapuram, Kerala 695014 (India); Siegel, Gene [Nanostructured Materials Research Laboratory, Department of Materials Science and Engineering, University of Utah, Salt Lake City, UT 84112 (United States); Tiwari, Ashutosh, E-mail: tiwari@eng.utah.edu [Nanostructured Materials Research Laboratory, Department of Materials Science and Engineering, University of Utah, Salt Lake City, UT 84112 (United States)

2015-01-01

A novel electrochemical glucose sensor was developed by employing a composite film of plant-like Zinc oxide (ZnO) and chitosan stabilized spherical gold nanoparticles (AuNPs) on which Glucose oxidaze (GOx) was immobilized. The ZnO was deposited on an indium tin oxide (ITO) coated glass and the AuNPs of average diameter of 23 nm were loaded on ZnO as the second layer. The prepared ITO/ZnO/AuNPs/GOx bioelectrode exhibited a low value of Michaelis–Menten constant of 1.70 mM indicating a good bio-matrix for GOx. The studies of electrochemical properties of the electrode using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) showed that, the presence of AuNPs provides significant enhancement of the electron transfer rate during redox reactions. The linear sweep voltammetry (LSV) shows that the ITO/ZnO/AuNPs/GOx based sensor has a high sensitivity of 3.12 μA·mM{sup −1}·cm{sup −2} in the range of 50 mg/dL to 400 mg/dL glucose concentration. The results show promising application of the gold nanoparticle modified plant-like ZnO composite bioelectrode for electrochemical sensing of glucose.

Determination of Glucose Concentration in Yeast Culture Medium

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Hara, Seiichi; Kishimoto, Tomokazu; Muraji, Masafumi; Tsujimoto, Hiroaki; Azuma, Masayuki; Ooshima, Hiroshi

The present paper describes a sensor for measuring the glucose concentration of yeast culture medium. The sensor determines glucose concentration by measuring the yield of hydrogen peroxide produced by glucose oxidase, which is monitored as luminescence using photomultiplier. The present sensor is able to measure low glucose concentration in media in which yeast cells keep respiration state. We herein describe the system and the characteristics of the glucose sensor.

Engineering the yeast Yarrowia lipolytica for the production of therapeutic proteins homogeneously glycosylated with Man8GlcNAc2 and Man5GlcNAc2

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De Pourcq Karen

2012-05-01

Full Text Available Abstract Background Protein-based therapeutics represent the fastest growing class of compounds in the pharmaceutical industry. This has created an increasing demand for powerful expression systems. Yeast systems are widely used, convenient and cost-effective. Yarrowia lipolytica is a suitable host that is generally regarded as safe (GRAS. Yeasts, however, modify their glycoproteins with heterogeneous glycans containing mainly mannoses, which complicates downstream processing and often interferes with protein function in man. Our aim was to glyco-engineer Y. lipolytica to abolish the heterogeneous, yeast-specific glycosylation and to obtain homogeneous human high-mannose type glycosylation. Results We engineered Y. lipolytica to produce homogeneous human-type terminal-mannose glycosylated proteins, i.e. glycosylated with Man8GlcNAc2 or Man5GlcNAc2. First, we inactivated the yeast-specific Golgi α-1,6-mannosyltransferases YlOch1p and YlMnn9p; the former inactivation yielded a strain producing homogeneous Man8GlcNAc2 glycoproteins. We tested this strain by expressing glucocerebrosidase and found that the hypermannosylation-related heterogeneity was eliminated. Furthermore, detailed analysis of N-glycans showed that YlOch1p and YlMnn9p, despite some initial uncertainty about their function, are most likely the α-1,6-mannosyltransferases responsible for the addition of the first and second mannose residue, respectively, to the glycan backbone. Second, introduction of an ER-retained α-1,2-mannosidase yielded a strain producing proteins homogeneously glycosylated with Man5GlcNAc2. The use of the endogenous LIP2pre signal sequence and codon optimization greatly improved the efficiency of this enzyme. Conclusions We generated a Y. lipolytica expression platform for the production of heterologous glycoproteins that are homogenously glycosylated with either Man8GlcNAc2 or Man5GlcNAc2 N-glycans. This platform expands the utility of Y. lipolytica as a

Naked-Eye Detection of Glucose in Saliva with Bienzymatic Paper-Based Sensor

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Luis A. Santana-Jiménez

2018-04-01

Full Text Available The high incidence of Diabetes Mellitus in low-income regions has promoted the development of low-cost alternatives to replace blood-based procedures. In this work, we present a bienzymatic paper-based sensor suitable for the naked-eye detection of glucose in saliva samples. The sensor was obtained by a stamping procedure and modified with chitosan to improve the colorimetric readout. The bienzymatic reaction of GOx-HRP coupled with 2,4,6-tribromo-3-hydroxy benzoic acid was applied for the detection of glucose within a range from 0 to 180 mgdL−1 in buffer and artificial saliva solutions. The visual readout was perceived by the naked eye and registered with an office scanner to evaluate the analytical performance. The results showed a limit of detection of 0.37 mgdL−1 (S/N = 3 with an R.S.D. of 1.69% and a linear range from 1 to 22.5 mgdL−1 with an R2 of 0.99235. The analysis of human saliva samples was performed without pre-processing, achieving recoveries from 92 to 114%. The naked-eye detection was evaluated under two different light settings, showing average recoveries of 108.58 and 90.65% for standard and low illumination. The proposed device showed potential for easy-to-use, sensitive, low-cost, fast, and device-free detection of salivary glucose suitable for untrained personnel operation and limited facilities.

Measuring O-GlcNAc cleavage by OGA and cell lysates on a peptide microarray.

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Sharif, Suhela; Shi, Jie; Bourakba, Mostafa; Ruijtenbeek, Rob; Pieters, Roland J

2017-09-01

O-GlcNAcylation is a post-translational modification resulting from the addition of an N-acetylglucosamine moiety to the hydroxyl groups of serine and threonine residues of nuclear and cytoplasmic proteins. In addition, O-GlcNAcylated proteins can be phosphorylated, which suggests the possibility for crosstalk between O-GlcNAcylation and phosphorylation. Dysregulation of O-GlcNAcylation affects cell signaling, transcriptional regulation, cell cycle control and can e.g. lead to tumorigenesis and tumor metastasis. There is a strong demand for efficient analytical techniques to better detect and investigate this abundant modification and its role in cancer. Herein we demonstrated the utility of an O-GlcNAcylated peptide array to examine O-GlcNAcase (OGA) activity and substrate specificity of both purified protein as well cell lysates of different cancer cell lines. Using this microarray, we clearly observed OGA activity and also inhibition thereof by OGA inhibitor thiamet G. Interestingly, different levels of OGA activity were observed of lysates derived from different cancer cell lines. This suggests that the tool may be useful in cancer research and biomarker development. Copyright © 2017 Elsevier Inc. All rights reserved.

In utero glucocorticoid (GLC) exposure reduces fetal skeletal muscle growth in rats

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Maternal undernutrition and stress expose the fetus to above normal levels of GLC and predispose to intrauterine growth restriction. The aim of this study was to determine if fetal GLC exposure impairs skeletal muscle growth independently of maternal undernutrition. Three groups (n=7/group) of timed...

Glucose ingestion during endurance training in men attenuates expression of myokine receptor

DEFF Research Database (Denmark)

Åkerström, Thorbjörn; Krogh-Madsen, Rikke; Petersen, Anne Marie Winther

2009-01-01

-leg) while ingesting a glucose solution (Glc) and ingested a placebo (Plc) while training the other leg (Plc-leg). Endurance training increased peak power by 14% and reduced the exercise-induced gene expression of IL-6 and IL-6Ralpha in skeletal muscle and IL-6 plasma concentration. The IL-6Ralpha density...

A glucose concentration and temperature sensor based on long period fiber gratings induced by electric-arc discharge

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Du, Chao; Wang, Qi

2017-10-01

As one of the key parameters in biological and chemical reactions, glucose concentration objectively reflects the characteristics of reactions, so the real-time monitoring of glucose concentration is important in the field of biochemical. Meanwhile, the influence from temperature should be considered. The fiber sensors have been studied extensively for decades due to the advantages of small size, immunity to electromagnetic interference and high sensitivity, which are suitable for the application of biochemical sensing. A long period fiber grating (LPFG) sensor induced by electric-arc discharge has been fabricated and demonstrated for simultaneous measurement of glucose concentration and temperature. The proposed sensor was fabricated by inscribing a sing mode fiber (SMF) with periodic electric-arc discharge technology. During the fabrication process, the electric-arc discharge technology was produced by a commercial fusion splicer, and the period of inscribed LPFG was determined by the movement of translation stages. A serials of periodic geometrical deformations would be formed in SMF after the fabrication, and the discharge intensity and discharge time can be adjusted though the fusion splicer settings screen. The core mode can be coupled into the cladding modes at certain wavelength when they satisfy the phase-matching conditions, and there will be several resonance dips in the transmission spectrum in LPFG. The resonance dips formed by the coupling between cladding modes and core mode have different sensitivity responses, so the simultaneous measurement for multi-parameter can be realized by monitoring the wavelength shifts of the resonance dips. Compared with the LPFG based on conventional SMF, the glucose concentration sensitivity has been obviously enhanced by etching the cladding with hydrofluoric acid solution. Based on the independent measured results, a dual-parameter measurement matrix has been built for signal demodulation. Because of the easy

Intraperitoneal Glucose Sensing is Sometimes Surprisingly Rapid

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Anders Lyngvi Fougner

2016-04-01

Full Text Available Rapid, accurate and robust glucose measurements are needed to make a safe artificial pancreas for the treatment of diabetes mellitus type 1 and 2. The present gold standard of continuous glucose sensing, subcutaneous (SC glucose sensing, has been claimed to have slow response and poor robustness towards local tissue changes such as mechanical pressure, temperature changes, etc. The present study aimed at quantifying glucose dynamics from central circulation to intraperitoneal (IP sensor sites, as an alternative to the SC location. Intraarterial (IA and IP sensors were tested in three anaesthetized non-diabetic pigs during experiments with intravenous infusion of glucose boluses, enforcing rapid glucose level excursions in the range 70--360 mg/dL (approximately 3.8--20 mmol/L. Optical interferometric sensors were used for IA and IP measurements. A first-order dynamic model with time delay was fitted to the data after compensating for sensor dynamics. Additionally, off-the-shelf Medtronic Enlite sensors were used for illustration of SC glucose sensing. The time delay in glucose excursions from central circulation (IA to IP sensor location was found to be in the range 0--26 s (median: 8.5 s, mean: 9.7 s, SD 9.5 s, and the time constant was found to be 0.5--10.2 min (median: 4.8 min, mean: 4.7 min, SD 2.9 min. IP glucose sensing sites have a substantially faster and more distinctive response than SC sites when sensor dynamics is ignored, and the peritoneal fluid reacts even faster to changes in intravascular glucose levels than reported in previous animal studies. This study may provide a benchmark for future, rapid IP glucose sensors.

AtRH57, a DEAD-box RNA helicase, is involved in feedback inhibition of glucose-mediated abscisic acid accumulation during seedling development and additively affects pre-ribosomal RNA processing with high glucose.

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Hsu, Yi-Feng; Chen, Yun-Chu; Hsiao, Yu-Chun; Wang, Bing-Jyun; Lin, Shih-Yun; Cheng, Wan-Hsing; Jauh, Guang-Yuh; Harada, John J; Wang, Co-Shine

2014-01-01

The Arabidopsis thaliana T-DNA insertion mutant rh57-1 exhibited hypersensitivity to glucose (Glc) and abscisic acid (ABA). The other two rh57 mutants also showed Glc hypersensitivity similar to rh57-1, strongly suggesting that the Glc-hypersensitive feature of these mutants results from mutation of AtRH57. rh57-1 and rh57-3 displayed severely impaired seedling growth when grown in Glc concentrations higher than 3%. The gene, AtRH57 (At3g09720), was expressed in all Arabidopsis organs and its transcript was significantly induced by ABA, high Glc and salt. The new AtRH57 belongs to class II DEAD-box RNA helicase gene family. Transient expression of AtRH57-EGFP (enhanced green fluorescent protein) in onion cells indicated that AtRH57 was localized in the nucleus and nucleolus. Purified AtRH57-His protein was shown to unwind double-stranded RNA independent of ATP in vitro. The ABA biosynthesis inhibitor fluridone profoundly redeemed seedling growth arrest mediated by sugar. rh57-1 showed increased ABA levels when exposed to high Glc. Quantitative real time polymerase chain reaction analysis showed that AtRH57 acts in a signaling network downstream of HXK1. A feedback inhibition of ABA accumulation mediated by AtRH57 exists within the sugar-mediated ABA signaling. AtRH57 mutation and high Glc conditions additively caused a severe defect in small ribosomal subunit formation. The accumulation of abnormal pre-rRNA and resistance to protein synthesis-related antibiotics were observed in rh57 mutants and in the wild-type Col-0 under high Glc conditions. These results suggested that AtRH57 plays an important role in rRNA biogenesis in Arabidopsis and participates in response to sugar involving Glc- and ABA signaling during germination and seedling growth. © 2013 The Authors The Plant Journal © 2013 John Wiley & Sons Ltd.

The ethylene glycol template assisted hydrothermal synthesis of Co3O4 nanowires; structural characterization and their application as glucose non-enzymatic sensor

International Nuclear Information System (INIS)

Khun, K.; Ibupoto, Z.H.; Liu, X.; Beni, V.; Willander, M.

2015-01-01

Highlights: • Ethylene glycol assisted Co 3 O 4 nanowires were synthesized by hydrothermal method. • The grown Co 3 O 4 nanowires were used for sensitive non-enzymatic glucose sensor. • The proposed glucose sensor shows a wide linear range with fast response. • The Co 3 O 4 modified electrode is a highly specific enzyme-less glucose sensor. - Abstract: In the work reported herein the ethylene glycol template assisted hydrothermal synthesis, onto Au substrate, of thin and highly dense cobalt oxide (Co 3 O 4 ) nanowires and their characterization and their application for non-enzymatic glucose sensing are reported. The structure and composition of Co 3 O 4 nanowires have been fully characterized using scanning electron microscopy, X-ray diffraction, high resolution transmission electron microscopy and X-ray photoelectron spectroscopy. The synthesized Co 3 O 4 nanowires resulted to have high purity and showed diameter of approximately 10 nm. The prepared Co 3 O 4 nanowires coated gold electrodes were applied to the non-enzymatic detection of glucose. The developed sensor showed high sensitivity (4.58 × 10 1 μA mM −1 cm −2 ), a wide linear range of concentration (1.00 × 10 −4 –1.2 × 10 1 mM) and a detection limit of 2.65 × 10 −5 mM. The developed glucose sensor has also shown to be very stable and selective over interferents such as uric acid and ascorbic acid. Furthermore, the proposed fabrication process was shown to be highly reproducible response (over nine electrodes)

Aralia elata inhibits neurodegeneration by downregulating O-GlcNAcylation of NF-κB in diabetic mice

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Seong-Jae Kim

2017-08-01

Full Text Available AIM: To investigate the role of O-GlcNAcylation of nuclear factor-kappa B (NF-κB in retinal ganglion cell (RGC death and analysedthe effect of Aralia elata (AE on neurodegeneration in diabetic mice. METHODS: C57BL/6mice with streptozotocin-induced diabetes were fed daily with AE extract or control (CTL diet at the onset of diabetes mellitus (DM. Two months after injection of streptozotocin or saline, the degree of cell death and the expression of O-GlcNAc transferase (OGT, N-acetyl-b-D-glucosaminidase (OGA, O-GlcNAcylated proteins, and O-GlcNAcylation of NF-κB were examined. RESULTS: AE did not affect the metabolic status of diabetic mice. The decrease in the inner retinal thickness (P<0.001 vs CTL, P<0.01 vs DM and increases in RGCs with terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (P<0.001 vs CTL, P<0.0001 vs DM, glial activation, and active caspase-3 (P<0.0001 vs CTL, P<0.0001 vs DM were blocked in diabetic retinas of AE extract-fed mice. Expression levels of protein O-GlcNAcylation and OGT were increased in diabetic retinas (P<0.0001 vs CTL, and the level of O-GlcNAcylation of the NF-κB p65 subunit was higher in diabetic retinas than in controls (P<0.0001 vs CTL. AE extract downregulated O-GlcNAcylation of NF-κB and prevented neurodegeneration induced by hyperglycemia (P<0.0001 vs DM. CONCLUSION: O-GlcNAcylation of NF-κB is concerned in neuronal degeneration and that AE prevents diabetes-induced RGC apoptosis via downregulation of NF-κB O-GlcNAcylation. Hence, O-GlcNAcylation may be a new object for the treatment of DR, and AE may have therapeutic possibility to prevent diabetes-induced neurodegeneration.

Spherulitic copper–copper oxide nanostructure-based highly sensitive nonenzymatic glucose sensor

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Das G

2015-08-01

Full Text Available Gautam Das, Thao Quynh Ngan Tran, Hyon Hee Yoon Department of Chemical and Biological Engineering, Gachon University, Seongnam, Republic of South Korea Abstract: In this work, three different spherulitic nanostructures Cu–CuOA, Cu–CuOB, and Cu–CuOC were synthesized in water-in-oil microemulsions by varying the surfactant concentration (30 mM, 40 mM, and 50 mM, respectively. The structural and morphological characteristics of the Cu–CuO nanostructures were investigated by ultraviolet–visible (UV–vis spectroscopy, X-ray diffraction, scanning electron microscopy, and high-resolution transmission electron microscopy techniques. The synthesized nanostructures were deposited on multiwalled carbon nanotube (MWCNT-modified indium tin oxide (ITO electrodes to fabricate a nonenzymatic highly sensitive amperometric glucose sensor. The performance of the ITO/MWCNT/Cu–CuO electrodes in the glucose assay was examined by cyclic voltammetry and chronoamperometric studies. The sensitivity of the sensor varied with the spherulite type; Cu–CuOA, Cu–CuOB, and Cu–CuOC exhibited a sensitivity of 1,229, 3,012, and 3,642 µA mM-1·cm-2, respectively. Moreover, the linear range is dependent on the structure types: 0.023–0.29 mM, 0.07–0.8 mM, and 0.023–0.34 mM for Cu–CuOA, Cu–CuOB, and Cu–CuOC, respectively. An excellent response time of 3 seconds and a low detection limit of 2 µM were observed for Cu–CuOB at an applied potential of +0.34 V. In addition, this electrode was found to be resistant to interference by common interfering agents such as urea, cystamine, l-ascorbic acid, and creatinine. The high performance of the Cu–CuO spherulites with nanowire-to-nanorod outgrowths was primarily due to the high surface area and stability, and good three-dimensional structure. Furthermore, the ITO/MWCNT/Cu–CuOB electrode applied to real urine and serum sample showed satisfactory performance. Keywords: copper oxide, multiwalled

Hydrogen peroxide and glucose concentration measurement using optical fiber grating sensors with corrodible plasmonic nanocoatings.

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Zhang, Xuejun; Wu, Ze; Liu, Fu; Fu, Qiangqiang; Chen, Xiaoyong; Xu, Jian; Zhang, Zhaochuan; Huang, Yunyun; Tang, Yong; Guo, Tuan; Albert, Jacques

2018-04-01

We propose and demonstrate hydrogen peroxide (H 2 O 2 ) and glucose concentration measurements using a plasmonic optical fiber sensor. The sensor utilizes a tilted fiber Bragg grating (TFBG) written in standard single mode communication fiber. The fiber is over coated with an nm-scale film of silver that supports surface plasmon resonances (SPRs). Such a tilted grating SPR structure provides a high density of narrow spectral resonances (Q-factor about 10 5 ) that overlap with the broader absorption band of the surface plasmon waves in the silver film, thereby providing an accurate tool to measure small shifts of the plasmon resonance frequencies. The H 2 O 2 to be detected acts as an oxidant to etch the silver film, which has the effect of gradually decreasing the SPR attenuation. The etching rate of the silver film shows a clear relationship with the H 2 O 2 concentration so that monitoring the progressively increasing attenuation of a selected surface plasmon resonance over a few minutes enables us to measure the H 2 O 2 concentration with a limit of detection of 0.2 μM. Furthermore, the proposed method can be applied to the determination of glucose in human serum for a concentration range from 0 to 12 mM (within the physiological range of 3-8 mM) by monitoring the H 2 O 2 produced by an enzymatic oxidation process. The sensor does not require accurate temperature control because of the inherent temperature insensitivity of TFBG devices referenced to the core mode resonance. A gold mirror coated on the fiber allows the sensor to work in reflection, which will facilitate the integration of the sensor with a hypodermic needle for in vitro measurements. The present study shows that Ag-coated TFBG-SPR can be applied as a promising type of sensing probe for optical detection of H 2 O 2 and glucose detection in human serum.

Hydrogen peroxide and glucose concentration measurement using optical fiber grating sensors with corrodible plasmonic nanocoatings

Science.gov (United States)

Zhang, Xuejun; Wu, Ze; Liu, Fu; Fu, Qiangqiang; Chen, Xiaoyong; Xu, Jian; Zhang, Zhaochuan; Huang, Yunyun; Tang, Yong; Guo, Tuan; Albert, Jacques

2018-01-01

We propose and demonstrate hydrogen peroxide (H2O2) and glucose concentration measurements using a plasmonic optical fiber sensor. The sensor utilizes a tilted fiber Bragg grating (TFBG) written in standard single mode communication fiber. The fiber is over coated with an nm-scale film of silver that supports surface plasmon resonances (SPRs). Such a tilted grating SPR structure provides a high density of narrow spectral resonances (Q-factor about 105) that overlap with the broader absorption band of the surface plasmon waves in the silver film, thereby providing an accurate tool to measure small shifts of the plasmon resonance frequencies. The H2O2 to be detected acts as an oxidant to etch the silver film, which has the effect of gradually decreasing the SPR attenuation. The etching rate of the silver film shows a clear relationship with the H2O2 concentration so that monitoring the progressively increasing attenuation of a selected surface plasmon resonance over a few minutes enables us to measure the H2O2 concentration with a limit of detection of 0.2 μM. Furthermore, the proposed method can be applied to the determination of glucose in human serum for a concentration range from 0 to 12 mM (within the physiological range of 3-8 mM) by monitoring the H2O2 produced by an enzymatic oxidation process. The sensor does not require accurate temperature control because of the inherent temperature insensitivity of TFBG devices referenced to the core mode resonance. A gold mirror coated on the fiber allows the sensor to work in reflection, which will facilitate the integration of the sensor with a hypodermic needle for in vitro measurements. The present study shows that Ag-coated TFBG-SPR can be applied as a promising type of sensing probe for optical detection of H2O2 and glucose detection in human serum. PMID:29675315

File list: His.PSC.05.H2BS112GlcNAc.AllCell [Chip-atlas[Archive

Lifescience Database Archive (English)

Full Text Available His.PSC.05.H2BS112GlcNAc.AllCell hg19 Histone H2BS112GlcNAc Pluripotent stem cell h...ttp://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.PSC.05.H2BS112GlcNAc.AllCell.bed ...

N-doped graphene-carbon nanotube hybrid networks attaching with gold nanoparticles for glucose non-enzymatic sensor.

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Jeong, Hun; Nguyen, Dang Mao; Lee, Min Sang; Kim, Hong Gun; Ko, Sang Cheol; Kwac, Lee Ku

2018-09-01

Herein, we successfully developed a novel three dimensional (3D) opened networks based on nitrogen doped graphene‑carbon nanotubes attaching with gold nanoparticles (N-GR-CNTs/AuNPs) to apply for non-enzymatic glucose determination. It was demonstrated that the N-GR-CNTs/AuNPs modified electrode exhibited good behavior for glucose detection with a long linear range of 2 μM to 19.6 mM, high sensitivity of 0.9824 μA·mM -1 ·cm -2 , low detection limit of 500 nM, and negligible interference effect. The high performance of the N-GR-CNTs/AuNPs based sensor was assumed due to the outstanding catalytic activity of AuNPs well dispersing on N-GR-CNTs networks, which exhibited as a perfect supporting scaffold due to the enhanced electrical conductivity and large surface area. The obtained results indicated that the N-GR-CNTs/AuNPs hybrid is highly promising for sensitive and selective detection of glucose in sensor application. Copyright © 2018 Elsevier B.V. All rights reserved.

Sodium Glucose Cotransporter 2 (SGLT2 Plays as a Physiological Glucose Sensor and Regulates Cellular Contractility in Rat Mesangial Cells.

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Masanori Wakisaka

Full Text Available Mesangial cells play an important role in regulating glomerular filtration by altering their cellular tone. We report the presence of a sodium glucose cotransporter (SGLT in rat mesangial cells. This study in rat mesangial cells aimed to evaluate the expression and role of SGLT2.The SGLT2 expression in rat mesangial cells was assessed by Western blotting and reverse transcription-polymerase chain reaction (RT-PCR. Changes in the mesangial cell surface area at different glucose concentrations and the effects of extracellular Na+ and Ca2+ and of SGLT and Na+/Ca2+ exchanger (NCX inhibitors on cellular size were determined. The cellular sizes and the contractile response were examined during a 6-day incubation with high glucose with or without phlorizin, an SGLT inhibitor.Western blotting revealed an SGLT2 band, and RT-PCR analysis of SGLT2 revealed the predicted 422-bp band in both rat mesangial and renal proximal tubular epithelial cells. The cell surface area changed according to the extracellular glucose concentration. The glucose-induced contraction was abolished by the absence of either extracellular Na+ or Ca2+ and by SGLT and NCX inhibitors. Under the high glucose condition, the cell size decreased for 2 days and increased afterwards; these cells did not contract in response to angiotensin II, and the SGLT inhibitor restored the abolished contraction.These data suggest that SGLT2 is expressed in rat mesangial cells, acts as a normal physiological glucose sensor and regulates cellular contractility in rat mesangial cells.

A correlation between altered O-GlcNAcylation, migration and with changes in E-cadherin levels in ovarian cancer cells

International Nuclear Information System (INIS)

Jin, Feng-zhen; Yu, Chao; Zhao, De-zhang; Wu, Ming-jun; Yang, Zhu

2013-01-01

O-GlcNAcylation is a dynamic and reversible posttranslational modification of nuclear and cytoplasmic proteins. In recent years, the roles of O-GlcNAcylation in several human malignant tumors have been investigated, and O-GlcNAcylation was found to be linked to cellular features relevant to metastasis. In this study, we modeled four diverse ovarian cancer cells and investigated the effects of O-GlcNAcylation on ovarian cancer cell migration. We found that total O-GlcNAcylation level was elevated in HO-8910PM cells compared to OVCAR3 cells. Additionally, through altering the total O-GlcNAcylation level by OGT silencing or OGA inhibition, we found that the migration of OVCAR3 cells was dramatically enhanced by PUGNAc and Thiamet G treatment, and the migration ability of HO-8910PM cells was significantly inhibited by OGT silencing. Furthermore, we also found that the expression of E-cadherin, an O-GlcNAcylated protein in ovarian cancer cells, was reduced by OGA inhibition in OVCAR3 cells and elevated by OGT silencing in HO-8910PM cells. These results indicate that O-GlcNAcylation could enhance ovarian cancer cell migration and decrease the expression of E-cadherin. Our studies also suggest that O-GlcNAcylation might become another potential target for the therapy of ovarian cancer. -- Highlights: • We examine the migration potential of diverse ovarian cancer cells. • We examine the total O-GlcNAcylation level of diverse ovarian cancer cells. • Increasing O-GlcNAcylation level will enhance the migration of ovarian cancer cells. • Reducing O-GlcNAcylation level will inhibit the migration of ovarian cancer cells. • The mechanism explains O-GlcNAcylation enhance ovarian cancer cell migration

Label-free electrochemical biosensing of small-molecule inhibition on O-GlcNAc glycosylation.

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Yang, Yu; Gu, Yuxin; Wan, Bin; Ren, Xiaomin; Guo, Liang-Hong

2017-09-15

O-linked N-acetylglucosamine (O-GlcNAc) transferase (OGT) plays a critical role in modulating protein function in many cellular processes and human diseases such as Alzheimer's disease and type II diabetes, and has emerged as a promising new target. Specific inhibitors of OGT could be valuable tools to probe the biological functions of O-GlcNAcylation, but a lack of robust nonradiometric assay strategies to detect glycosylation, has impeded efforts to identify such compounds. Here we have developed a novel label-free electrochemical biosensor for the detection of peptide O-GlcNAcylation using protease-protection strategy and electrocatalytic oxidation of tyrosine mediated by osmium bipyridine as a signal reporter. There is a large difference in the abilities of proteolysis of the glycosylated and the unglycosylated peptides by protease, thus providing a sensing mechanism for OGT activity. When the O-GlcNAcylation is achieved, the glycosylated peptides cannot be cleaved by proteinase K and result in a high current response on indium tin oxide (ITO) electrode. However, when the O-GlcNAcylation is successfully inhibited using a small molecule, the unglycosylated peptides can be cleaved easily and lead to low current signal. Peptide O-GlcNAcylation reaction was performed in the presence of a well-defined small-molecule OGT inhibitor. The results indicated that the biosensor could be used to screen the OGT inhibitors effectively. Our label-free electrochemical method is a promising candidate for protein glycosylation pathway research in screening small-molecule inhibitors of OGT. Copyright © 2017 Elsevier B.V. All rights reserved.

Non-invasive Blood Glucose Quantification Using a Hybrid Sensor

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Sundararajan JAYAPAL

2009-02-01

Full Text Available Diabetes Mellitus is a group of metabolic diseases characterized by high blood sugar (glucose levels which result from defects in insulin secretion. It is very important for the diabetics and normal people to have a correct blood glucose level. The HbA1c test is the most preferred test by renowned doctors for glucose quantification. But this test is an invasive one. At present, there are many available techniques for this purpose but these are mostly invasive or minimally non-invasive and most of these are under research. Among the different methods available, the photo acoustic (PA methods provide a reliable solution since the acoustical energy loss is much less compared to the optical or other techniques. Here a novel framework is presented for blood glucose level measurement using a combination of the HbA1c test and a PA method to get an absolutely consistent and precise, non-invasive technique. The setup uses a pulsed laser diode with pulse duration of 5-15 ns and at a repetition rate of 10 Hz as the source. The detector setup is based on the piezoelectric detection. It consists of a ring detector that includes two double ring sensors that are attached to the ring shaped module that can be worn around the finger. The major aim is to detect the photo acoustic signals from the glycated hemoglobin with the least possible error. The proposed monitoring system is designed with extreme consideration to precision and compatibility with the other computing devices. The results obtained in this research have been studied and analyzed by comparing these with those of in-vitro techniques like the HPLC. The comparison has been plotted and it shows a least error. The results also show a positive drive for using this concept as a basis for future extension in quantifying the other blood components.

Soft, smart contact lenses with integrations of wireless circuits, glucose sensors, and displays.

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Park, Jihun; Kim, Joohee; Kim, So-Yun; Cheong, Woon Hyung; Jang, Jiuk; Park, Young-Geun; Na, Kyungmin; Kim, Yun-Tae; Heo, Jun Hyuk; Lee, Chang Young; Lee, Jung Heon; Bien, Franklin; Park, Jang-Ung

2018-01-01

Recent advances in wearable electronics combined with wireless communications are essential to the realization of medical applications through health monitoring technologies. For example, a smart contact lens, which is capable of monitoring the physiological information of the eye and tear fluid, could provide real-time, noninvasive medical diagnostics. However, previous reports concerning the smart contact lens have indicated that opaque and brittle components have been used to enable the operation of the electronic device, and this could block the user's vision and potentially damage the eye. In addition, the use of expensive and bulky equipment to measure signals from the contact lens sensors could interfere with the user's external activities. Thus, we report an unconventional approach for the fabrication of a soft, smart contact lens in which glucose sensors, wireless power transfer circuits, and display pixels to visualize sensing signals in real time are fully integrated using transparent and stretchable nanostructures. The integration of this display into the smart lens eliminates the need for additional, bulky measurement equipment. This soft, smart contact lens can be transparent, providing a clear view by matching the refractive indices of its locally patterned areas. The resulting soft, smart contact lens provides real-time, wireless operation, and there are in vivo tests to monitor the glucose concentration in tears (suitable for determining the fasting glucose level in the tears of diabetic patients) and, simultaneously, to provide sensing results through the contact lens display.

Protein O-GlcNAc Modification Increases in White Blood Cells After a Single Bout of Physical Exercise.

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Nagy, Tamás; Kátai, Emese; Fisi, Viktória; Takács, Tamás Tibor; Stréda, Antal; Wittmann, István; Miseta, Attila

2018-01-01

Protein O-linked N -acetylglucosamine (O-GlcNAc) is a dynamic posttranslational modification influencing the function of many intracellular proteins. Recently it was revealed that O-GlcNAc regulation is modified under various stress states, including ischemia and oxidative stress. Aside from a few contradictory studies based on animal models, the effect of exercise on O-GlcNAc is unexplored. To evaluate O-GlcNAc levels in white blood cells (WBC) of human volunteers following physical exercise. Young (age 30 ± 5.2), healthy male volunteers ( n  = 6) were enlisted for the study. Blood parameters including metabolites, ions, "necro"-enzymes, and cell counts were measured before and after a single bout of exercise (2-mile run). From WBC samples, we performed western blots to detect O-GlcNAc modified proteins. The distribution of O-GlcNAc in WBC subpopulations was assessed by flow cytometry. Elevation of serum lactic acid (increased from 1.3 ± 0.4 to 6.9 ± 1.7 mM), creatinine (from 77.5 ± 6.3 U/L to 102.2 ± 7.0 μM), and lactate dehydrogenase (from 318.5 ± 26.2 to 380.5 ± 33.2 U/L) confirmed the effect of exercise. WBC count also significantly increased (from 6.6 ± 1.0 to 8.4 ± 1.4 G/L). The level of O-GlcNAc modified proteins in WBCs showed significant elevation after exercise (85 ± 51%, p  O-GlcNAc status of WBCs. O-GlcNAc modification could be a natural process by which physical activity modulates the immune system. Further research could elucidate the role of O-GlcNAc during exercise and validate O-GlcNAc as a biomarker for fitness assessment.

Fabrication of Nonenzymatic Glucose Sensors Based on Multiwalled Carbon Nanotubes with Bimetallic Pt-M (M = Ru and Sn Catalysts by Radiolytic Deposition

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Sun-Young Kwon

2012-01-01

Full Text Available Nonenzymatic glucose sensors employing multiwalled carbon nanotubes (MWNTs with highly dispersed Pt-M (M = Ru and Sn nanoparticles (Pt-M@PVP-MWNTs were fabricated by radiolytic deposition. The Pt-M nanoparticles on the MWNTs were characterized by transmittance electron microscopy, elemental analysis, and X-ray diffraction. They were found to be well dispersed and to exhibit alloy properties on the MWNT support. Electrochemical testing showed that these nonenzymatic sensors had larger currents (mA than that of a bare glassy carbon (GC electrode and one modified with MWNTs. The sensitivity (A mM−1, linear range (mM, and detection limit (mM (S/N = 3 of the glucose sensor with the Pt-Ru catalyst in NaOH electrolyte were determined as 18.0, 1.0–2.5, 0.7, respectively. The corresponding data of the sensor with Pt-Sn catalyst were 889.0, 1.00–3.00, and 0.3, respectively. In addition, these non-enzymatic sensors can effectively avoid interference arising from the oxidation of the common interfering species ascorbic acid and uric acid in NaOH electrolyte. The experimental results show that such sensors can be applied in the detection of glucose in commercial red wine samples.

Role of O-GlcNAcylation in nutritional sensing, insulin resistance and in mediating the benefits of exercise.

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Myslicki, Jason P; Belke, Darrell D; Shearer, Jane

2014-11-01

The purpose of this review is to highlight the role of O-linked β-N-acetylglucosamine (O-GlcNAc) protein modification in metabolic disease states and to summarize current knowledge of how exercise affects this important post-translational signalling pathway. O-GlcNAc modification is an intracellular tool capable of integrating energy supply with demand. The accumulation of excess energy associated with obesity and insulin resistance is mediated, in part, by the hexosamine biosynthetic pathway (HBP), which results in the O-GlcNAcylation of a myriad of proteins, thereby affecting their respective function, stability, and localization. Insulin resistance is related to the excessive O-GlcNAcylation of key metabolic proteins causing a chronic blunting of insulin signalling pathways and precipitating the accompanying pathologies, such as heart and kidney disease. Lifestyle modifications such as diet and exercise also modify the pathway. Exercise is a front-line and cost-effective therapeutic approach for insulin resistance, and recent work shows that the intervention can alter O-GlcNAc gene expression, signalling, and protein modification. However, there is currently no consensus on the effect of frequency, intensity, type, and duration of exercise on O-GlcNAc modification, the HBP, and its related enzymes. On one end of the spectrum, mild, prolonged swim training reduces O-GlcNAcylation, while on the other end, higher intensity treadmill running increases cardiac protein O-GlcNAc modification. Clearly, a balance between acute and chronic stress of exercise is needed to reap the benefits of the intervention on O-GlcNAc signalling.

The glucuronyltransferase GlcAT-P is required for stretch growth of peripheral nerves in Drosophila.

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Pandey, Rahul; Blanco, Jorge; Udolph, Gerald

2011-01-01

During development, the growth of the animal body is accompanied by a concomitant elongation of the peripheral nerves, which requires the elongation of integrated nerve fibers and the axons projecting therein. Although this process is of fundamental importance to almost all organisms of the animal kingdom, very little is known about the mechanisms regulating this process. Here, we describe the identification and characterization of novel mutant alleles of GlcAT-P, the Drosophila ortholog of the mammalian glucuronyltransferase b3gat1. GlcAT-P mutants reveal shorter larval peripheral nerves and an elongated ventral nerve cord (VNC). We show that GlcAT-P is expressed in a subset of neurons in the central brain hemispheres, in some motoneurons of the ventral nerve cord as well as in central and peripheral nerve glia. We demonstrate that in GlcAT-P mutants the VNC is under tension of shorter peripheral nerves suggesting that the VNC elongates as a consequence of tension imparted by retarded peripheral nerve growth during larval development. We also provide evidence that for growth of peripheral nerve fibers GlcAT-P is critically required in hemocytes; however, glial cells are also important in this process. The glial specific repo gene acts as a modifier of GlcAT-P and loss or reduction of repo function in a GlcAT-P mutant background enhances VNC elongation. We propose a model in which hemocytes are required for aspects of glial cell biology which in turn affects the elongation of peripheral nerves during larval development. Our data also identifies GlcAT-P as a first candidate gene involved in growth of integrated peripheral nerves and therefore establishes Drosophila as an amenable in-vivo model system to study this process at the cellular and molecular level in more detail.

The glucuronyltransferase GlcAT-P is required for stretch growth of peripheral nerves in Drosophila.

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Rahul Pandey

Full Text Available During development, the growth of the animal body is accompanied by a concomitant elongation of the peripheral nerves, which requires the elongation of integrated nerve fibers and the axons projecting therein. Although this process is of fundamental importance to almost all organisms of the animal kingdom, very little is known about the mechanisms regulating this process. Here, we describe the identification and characterization of novel mutant alleles of GlcAT-P, the Drosophila ortholog of the mammalian glucuronyltransferase b3gat1. GlcAT-P mutants reveal shorter larval peripheral nerves and an elongated ventral nerve cord (VNC. We show that GlcAT-P is expressed in a subset of neurons in the central brain hemispheres, in some motoneurons of the ventral nerve cord as well as in central and peripheral nerve glia. We demonstrate that in GlcAT-P mutants the VNC is under tension of shorter peripheral nerves suggesting that the VNC elongates as a consequence of tension imparted by retarded peripheral nerve growth during larval development. We also provide evidence that for growth of peripheral nerve fibers GlcAT-P is critically required in hemocytes; however, glial cells are also important in this process. The glial specific repo gene acts as a modifier of GlcAT-P and loss or reduction of repo function in a GlcAT-P mutant background enhances VNC elongation. We propose a model in which hemocytes are required for aspects of glial cell biology which in turn affects the elongation of peripheral nerves during larval development. Our data also identifies GlcAT-P as a first candidate gene involved in growth of integrated peripheral nerves and therefore establishes Drosophila as an amenable in-vivo model system to study this process at the cellular and molecular level in more detail.

Determination of hydrogen peroxide and glucose using a novel sensor platform based on Co0.4Fe0.6LaO3 nanoparticles

International Nuclear Information System (INIS)

Zhang, Zhen; Gu, Shuqing; Ding, Yaping; Zhang, Fenfen; Jin, Jindi

2013-01-01

We report on a novel nonenzymatic sensor platform for the determination of hydrogen peroxide and glucose. It is based on a carbon paste electrode that was modified with Co 0.4 Fe 0.6 LaO 3 nanoparticles synthesized by the sol–gel method. The structure and morphology of Co 0.4 Fe 0.6 LaO 3 nanoparticles were characterized by X-ray diffraction, scanning electron microscopy, and transmission electron microscopy. The electrochemical performance of this sensor was evaluated by cyclic voltammetry and amperometry, and the results demonstrated that it exhibits strong electrocatalytical activity towards the oxidation of H 2 O 2 and glucose in an alkaline medium. The sensor has a limit of detection as low as 2.0 nM of H 2 O 2 and a linear range that extends from 0.01 to 800 μM. The response to glucose is characterized by two analytical ranges of different slope, viz. from 0.05 to 5 μM and from 5 to 500 μM, with a 10 nM limit of detection. The glucose sensor has a fast response and good long term stability. (author)

Absolute quantification of regional cerebral glucose utilization in mice by 18F-FDG small animal PET scanning and 2-14C-DG autoradiography.

Science.gov (United States)

Toyama, Hiroshi; Ichise, Masanori; Liow, Jeih-San; Modell, Kendra J; Vines, Douglass C; Esaki, Takanori; Cook, Michelle; Seidel, Jurgen; Sokoloff, Louis; Green, Michael V; Innis, Robert B

2004-08-01

The purpose of this study was to evaluate the feasibility of absolute quantification of regional cerebral glucose utilization (rCMR(glc)) in mice by use of (18)F-FDG and a small animal PET scanner. rCMR(glc) determined with (18)F-FDG PET was compared with values determined simultaneously by the autoradiographic 2-(14)C-DG method. In addition, we compared the rCMR(glc) values under isoflurane, ketamine and xylazine anesthesia, and awake states. Immediately after injection of (18)F-FDG and 2-(14)C-DG into mice, timed arterial samples were drawn over 45 min to determine the time courses of (18)F-FDG and 2-(14)C-DG. Animals were euthanized at 45 min and their brain was imaged with the PET scanner. The brains were then processed for 2-(14)C-DG autoradiography. Regions of interest were manually placed over cortical regions on corresponding coronal (18)F-FDG PET and 2-(14)C-DG autoradiographic images. rCMR(glc) values were calculated for both tracers by the autoradiographic 2-(14)C-DG method with modifications for the different rate and lumped constants for the 2 tracers. Average rCMR(glc) values in cerebral cortex with (18)F-FDG PET under normoglycemic conditions (isoflurane and awake) were generally lower (by 8.3%) but strongly correlated with those of 2-(14)C-DG (r(2) = 0.95). On the other hand, under hyperglycemic conditions (ketamine/xylazine) average cortical rCMR(glc) values with (18)F-FDG PET were higher (by 17.3%) than those with 2-(14)C-DG. Values for rCMR(glc) and uptake (percentage injected dose per gram [%ID/g]) with (18)F-FDG PET were significantly lower under both isoflurane and ketamine/xylazine anesthesia than in the awake mice. However, the reductions of rCMR(glc) were markedly greater under isoflurane (by 57%) than under ketamine and xylazine (by 19%), whereas more marked reductions of %ID/g were observed with ketamine/xylazine (by 54%) than with isoflurane (by 37%). These reverse differences between isoflurane and ketamine/xylazine may be due to

Development of Chemically Amplified Optical Sensors for Continuous Blood Glucose Monitoring Final Report CRADA No. TSB-1162-95

Energy Technology Data Exchange (ETDEWEB)

Lane, Stephen M. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Univ. of California, Livermore, CA (United States); Mastrototaro, John J. [Minimed Technologies, Inc., Sylmar, CA (United States)

2018-01-22

Diabetes is a chronic disease that affects 14 million people in the U.S. and more than 110 million people worldwide. Each year in this country 27,000 diabetic patients become blind, 15,000 have kidney failure, and over 54,000 have peripheral limb amputations. In 1992, total healthcare costs in the U.S. for diabetes were more than $105 billion, approximately 15% of our healthcare budget. Conventional therapy for the most severe form of diabetes, insulin-dependent diabetes mellitus (IDDM) or Type I diabetes, is to administer one or two injections per day of various forms of insulin while monitoring blood glucose levels twice or three times daily with commercial glucometers that require blood samples. Near normal blood sugar levels (glycemic control) is difficult to achieve with conventional therapy. In the fall of 1993, the results of the 10-year $165 million Diabetes Control and Complications Trial (DCCT) were published which showed that intensive insulin management would lead to dramatically fewer cases of retinopathy (which leads to blindness), nephropathy (which leads to kidney failure), and neuropathy (which can lead to limb amputations) [New England Journal of Medicine, Vo1239, No.14 977-986 (1993)]. If existing commercial insulin pumps could be combined with a continuous glucose sensor, a more physiological and fine-tuned therapy could be provided - in effect, an artificial biomechanical pancreas would be available. Existing research suggested that such a development would dramatically improve glucose control, thus greatly reducing morbidity and mortality from this disease. MiniMed Technologies in Sylmar, CA, identified a number of optically based sensor strategies as well as candidate chemical reactions that could be used to implement a minimally invasive opto-chemical glucose sensor. LLNL evaluated these sensor strategies and chemical reactions. These evaluations were the first steps leading to development of a sensor of considerable importance that could

O-GlcNAcPRED-II: an integrated classification algorithm for identifying O-GlcNAcylation sites based on fuzzy undersampling and a K-means PCA oversampling technique.

Science.gov (United States)

Jia, Cangzhi; Zuo, Yun; Zou, Quan; Hancock, John

2018-02-06

Protein O-GlcNAcylation (O-GlcNAc) is an important post-translational modification of serine (S)/threonine (T) residues that involves multiple molecular and cellular processes. Recent studies have suggested that abnormal O-G1cNAcylation causes many diseases, such as cancer and various neurodegenerative diseases. With the available protein O-G1cNAcylation sites experimentally verified, it is highly desired to develop automated methods to rapidly and effectively identify O-G1cNAcylation sites. Although some computational methods have been proposed, their performance has been unsatisfactory, particularly in terms of prediction sensitivity. In this study, we developed an ensemble model O-GlcNAcPRED-II to identify potential O-G1cNAcylation sites. A K-means principal component analysis oversampling technique (KPCA) and fuzzy undersampling method (FUS) were first proposed and incorporated to reduce the proportion of the original positive and negative training samples. Then, rotation forest, a type of classifier-integrated system, was adopted to divide the eight types of feature space into several subsets using four sub-classifiers: random forest, k-nearest neighbour, naive Bayesian and support vector machine. We observed that O-GlcNAcPRED-II achieved a sensitivity of 81.05%, specificity of 95.91%, accuracy of 91.43% and Matthew's correlation coefficient of 0.7928 for five-fold cross-validation run 10 times. Additionally, the results obtained by O-GlcNAcPRED-II on two independent datasets also indicated that the proposed predictor outperformed five published prediction tools. http://121.42.167.206/OGlcPred/. cangzhijia@dlmu.edu.cn or zouquan@nclab.net. © The Author (2018). Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com

OGT (O-GlcNAc Transferase) Selectively Modifies Multiple Residues Unique to Lamin A.

Science.gov (United States)

Simon, Dan N; Wriston, Amanda; Fan, Qiong; Shabanowitz, Jeffrey; Florwick, Alyssa; Dharmaraj, Tejas; Peterson, Sherket B; Gruenbaum, Yosef; Carlson, Cathrine R; Grønning-Wang, Line M; Hunt, Donald F; Wilson, Katherine L

2018-05-17

The LMNA gene encodes lamins A and C with key roles in nuclear structure, signaling, gene regulation, and genome integrity. Mutations in LMNA cause over 12 diseases ('laminopathies'). Lamins A and C are identical for their first 566 residues. However, they form separate filaments in vivo, with apparently distinct roles. We report that lamin A is β- O -linked N -acetylglucosamine- (O -GlcNAc)-modified in human hepatoma (Huh7) cells and in mouse liver. In vitro assays with purified O -GlcNAc transferase (OGT) enzyme showed robust O -GlcNAcylation of recombinant mature lamin A tails (residues 385⁻646), with no detectable modification of lamin B1, lamin C, or 'progerin' (Δ50) tails. Using mass spectrometry, we identified 11 O -GlcNAc sites in a 'sweet spot' unique to lamin A, with up to seven sugars per peptide. Most sites were unpredicted by current algorithms. Double-mutant (S612A/T643A) lamin A tails were still robustly O -GlcNAc-modified at seven sites. By contrast, O -GlcNAcylation was undetectable on tails bearing deletion Δ50, which causes Hutchinson⁻Gilford progeria syndrome, and greatly reduced by deletion Δ35. We conclude that residues deleted in progeria are required for substrate recognition and/or modification by OGT in vitro. Interestingly, deletion Δ35, which does not remove the majority of identified O -GlcNAc sites, does remove potential OGT-association motifs (lamin A residues 622⁻625 and 639⁻645) homologous to that in mouse Tet1. These biochemical results are significant because they identify a novel molecular pathway that may profoundly influence lamin A function. The hypothesis that lamin A is selectively regulated by OGT warrants future testing in vivo, along with two predictions: genetic variants may contribute to disease by perturbing OGT-dependent regulation, and nutrient or other stresses might cause OGT to misregulate wildtype lamin A.

Measurement of O-GlcNAcylated endothelial nitric oxide synthase by using 2',5'-ADP-Sepharose pull-down assay.

Science.gov (United States)

Long, Yang; Yan, Jianghong; Luo, Suxin; Liu, Zhenguo; Xia, Yong

2017-11-15

Endothelial nitric oxide synthase (eNOS) plays central roles in cardiovascular regulation and disease. eNOS function is critically affected by O-linked N-acetylglucosamine (O-GlcNAc) modification. The present method for measuring O-GlcNAcylated eNOS relies on immunoprecipitation. Such method exhibits low detection efficiency and is also costly. We here report a simplified assay by employing the high binding affinity of eNOS with the 2',5'-ADP-Sepharose resins. Together with the O-GlcNAc antibody, this assay readily allows the detection of O-GlcNAcylated eNOS in both cultured endothelial cells and rat vascular tissues. By using this assay, we demonstrate that eNOS O-GlcNAcylation is markedly elevated in the vessels of diabetic rats. Thus, a 2',5'-ADP-Sepharose-based pull-down assay is developed to measure O-GlcNAcylated eNOS. This assay is simple and efficient in detecting O-GlcNAcylated eNOS in cultured cells and animal tissues under both normal and disease conditions. Copyright © 2017 Elsevier Inc. All rights reserved.

Hijacking of the O-GlcNAcZYME complex by the HTLV-1 Tax oncoprotein facilitates viral transcription.

Science.gov (United States)

Groussaud, Damien; Khair, Mostafa; Tollenaere, Armelle I; Waast, Laetitia; Kuo, Mei-Shiue; Mangeney, Marianne; Martella, Christophe; Fardini, Yann; Coste, Solène; Souidi, Mouloud; Benit, Laurence; Pique, Claudine; Issad, Tarik

2017-07-01

The viral Tax oncoprotein plays a key role in both Human T-cell lymphotropic virus type 1 (HTLV-1)-replication and HTLV-1-associated pathologies, notably adult T-cell leukemia. Tax governs the transcription from the viral 5'LTR, enhancing thereby its own expression, via the recruitment of dimers of phosphorylated CREB to cAMP-response elements located within the U3 region (vCRE). In addition to phosphorylation, CREB is also the target of O-GlcNAcylation, another reversible post-translational modification involved in a wide range of diseases, including cancers. O-GlcNAcylation consists in the addition of O-linked-N-acetylglucosamine (O-GlcNAc) on Serine or Threonine residues, a process controlled by two enzymes: O-GlcNAc transferase (OGT), which transfers O-GlcNAc on proteins, and O-GlcNAcase (OGA), which removes it. In this study, we investigated the status of O-GlcNAcylation enzymes in HTLV-1-transformed T cells. We found that OGA mRNA and protein expression levels are increased in HTLV-1-transformed T cells as compared to control T cell lines while OGT expression is unchanged. However, higher OGA production coincides with a reduction in OGA specific activity, showing that HTLV-1-transformed T cells produce high level of a less active form of OGA. Introducing Tax into HEK-293T cells or Tax-negative HTLV-1-transformed TL-om1 T cells is sufficient to inhibit OGA activity and increase total O-GlcNAcylation, without any change in OGT activity. Furthermore, Tax interacts with the OGT/OGA complex and inhibits the activity of OGT-bound OGA. Pharmacological inhibition of OGA increases CREB O-GlcNAcylation as well as HTLV-1-LTR transactivation by Tax and CREB recruitment to the LTR. Moreover, overexpression of wild-type CREB but not a CREB protein mutated on a previously described O-GlcNAcylation site enhances Tax-mediated LTR transactivation. Finally, both OGT and OGA are recruited to the LTR. These findings reveal the interplay between Tax and the O-GlcNAcylation pathway

Hyper-O-GlcNAcylation of YB-1 affects Ser102 phosphorylation and promotes cell proliferation in hepatocellular carcinoma

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Liu, Qingqing [Department of Gastroenterology, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province (China); Jiangsu Province Key Laboratory for Inflammation and Molecular Drug Target, Nantong University, 19 Qi-xiu Road, Nantong 226001, Jiangsu Province (China); Tao, Tao [Jiangsu Province Key Laboratory for Inflammation and Molecular Drug Target, Nantong University, 19 Qi-xiu Road, Nantong 226001, Jiangsu Province (China); Liu, Fang [Key Laboratory of Neuroregeneration, Nantong University, Nantong 226001, Jiangsu Province (China); Ni, Runzhou [Department of Gastroenterology, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province (China); Lu, Cuihua, E-mail: lch1516@yeah.net [Department of Gastroenterology, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province (China); Shen, Aiguo, E-mail: shag@ntu.edu.cn [Jiangsu Province Key Laboratory for Inflammation and Molecular Drug Target, Nantong University, 19 Qi-xiu Road, Nantong 226001, Jiangsu Province (China); Key Laboratory of Neuroregeneration, Nantong University, Nantong 226001, Jiangsu Province (China)

2016-12-10

As an essential post-translational modification, O-GlcNAcylation has been thought to be able to modulate various nuclear and cytoplasmic proteins and is emerging as a key regulator of multiple biological processes, such as transcription, cell growth, signal transduction, and cell motility. Recently, authoritative glycomics analyses have reported extensive crosstalk between O-GlcNAcylation and phosphorylation, which always dynamically interplay with each other and regulate signaling, transcription, and other cellular processes. Also, plentiful studies have shown close correlation between YB-1 phosphorylation and tumorigenesis. Therefore, our study aimed to determine whether YB-1 was O-GlcNAc modified and whether such modification could interact with its phosphorylation during the process of HCC development. Western blot and immunohistochemistry were firstly conducted to reveal obvious up-regulation of YB-1, OGT and O-GlcNAc modification in HCC tissues. What is more, not only YB-1 was identified to be O-GlcNAcylated but hyper-O-GlcNAcylation was demonstrated to facilitate HCC cell proliferation in a YB-1 dependent manner. Moreover, we detected four specific O-GlcNAc sites and confirmed T126A to be the most effective mutant in HCC cell proliferation via close O-GlcNAcylation-phosphorylation interaction. Even more interestingly, we discovered that T126A-induced HCC cell retardation and subdued transcriptional activity of YB-1 could be partially reversed by T126A/S102E mutant. From all above, it is not difficult to find that glycosylated-YB-1 mainly enhanced cell proliferation through congenerous actions with YB-1 phosphorylation and thus played indispensable roles in fine-tuning cell proliferation and procession of HCC. - Highlights: • YB-1 and OGT are associated with HCC prognosis. • YB-1 is O-GlcNAc modified in HCC. • Hyper-O-GlcNAcylation promotes HCC cell proliferation in dependent of YB-1. • The proliferating role of O-GlcNAcylation is based on Ser102

The effect of O-GlcNAcylation on hnRNP A1 translocation and interaction with transportin1

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Roth, Shira; Khalaila, Isam, E-mail: isam@bgu.ac.il

2017-01-01

The heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) is a major pre-mRNA binding protein involved in transcription and translation. Although predominantly nuclear, hnRNP A1 shuttles rapidly between the nucleus and the cytosol, delivering its anchored pre-mRNA for further processing. Translocation is important for hnRNP A1 to accomplish its transcriptional and translational roles. Transportin1 (Trn1), a translocation protein, facilitates the translocation of hnRNP A1 back to the nucleus. Moreover, phosphorylation of serine residues at hnRNP A1 C-terminal domain affects its translocation. In this study, we found that phosphorylation is not the only modification that hnRNP A1 undergoes, but also O-linked N-acetylglucosaminylation (O-GlcNAcylation) could occur. Several putative novel O-GlcNAcylation and phosphorylation sites in hnRNP A1 were mapped. Whereas enhanced O-GlcNAcylation increased hnRNP A1 interaction with Trn1, enhanced phosphorylation reduced the interaction between the proteins. In addition, elevated O-GlcNAcylation resulted in hnRNP A1 seclusion in the nucleus, whereas elevated phosphorylation resulted in its accumulation in the cytosol. These findings suggest that a new player, i.e., O-GlcNAcylation, regulates hnRNP A1 translocation and interaction with Trn1, possibly affecting its function. There is a need for further study, to elucidate the role of O-GlcNAcylation in the regulation of the specific activities of hnRNP A1 in transcription and translation. - Highlights: • O-GlcNAcylation regulates hnRNP A1 translocation and interaction with Trn1. • Reciprocity between phosphorylation and O-GlcNAcylation in hnRNP A1 is proposed. • Novel O-GlcNAcylation and phosphorylation sites on hnRNPA1 were identified.

Interaction between O-GlcNAc modification and tyrosine phosphorylation of prohibitin: implication for a novel binary switch.

Directory of Open Access Journals (Sweden)

Sudharsana R Ande

Full Text Available Prohibitin (PHB or PHB1 is an evolutionarily conserved, multifunctional protein which is present in various cellular compartments including the plasma membrane. However, mechanisms involved in various functions of PHB are not fully explored yet. Here we report for the first time that PHB interacts with O-linked beta-N-acetylglucosamine transferase (O-GlcNAc transferase, OGT and is O-GlcNAc modified; and also undergoes tyrosine phosphorylation in response to insulin. Tyrosine 114 (Tyr114 and tyrosine 259 (Tyr259 in PHB are in the close proximity of potential O-GlcNAc sites serine 121 (Ser121 and threonine 258 (Thr258 respectively. Substitution of Tyr114 and Tyr259 residues in PHB with phenylalanine by site-directed mutagenesis results in reduced tyrosine phosphorylation as well as reduced O-GlcNAc modification of PHB. Surprisingly, this also resulted in enhanced tyrosine phosphorylation and activity of OGT. This is attributed to the presence of similar tyrosine motifs in PHB and OGT. Substitution of Ser121 and Thr258 with alanine and isoleucine respectively resulted in attenuation of O-GlcNAc modification and increased tyrosine phosphorylation of PHB suggesting an association between these two dynamic modifications. Sequence analysis of O-GlcNAc modified proteins having known O-GlcNAc modification site(s or known tyrosine phosphorylation site(s revealed a strong potential association between these two posttranslational modifications in various proteins. We speculate that O-GlcNAc modification and tyrosine phosphorylation of PHB play an important role in tyrosine kinase signaling pathways including insulin, growth factors and immune receptors signaling. In addition, we propose that O-GlcNAc modification and tyrosine phosphorylation is a novel previously unidentified binary switch which may provide new mechanistic insights into cell signaling pathways and is open for direct experimental examination.

O-GlcNAc modification of the coat protein of the potyvirus Plum pox virus enhances viral infection.

Science.gov (United States)

Pérez, José de Jesús; Udeshi, Namrata D; Shabanowitz, Jeffrey; Ciordia, Sergio; Juárez, Silvia; Scott, Cheryl L; Olszewski, Neil E; Hunt, Donald F; García, Juan Antonio

2013-08-01

O-GlcNAcylation is a dynamic protein modification which has been studied mainly in metazoans. We reported previously that an Arabidopsis thaliana O-GlcNAc transferase modifies at least two threonine residues of the Plum pox virus (PPV) capsid protein (CP). Now, six additional residues were shown to be involved in O-GlcNAc modification of PPV CP. CP O-GlcNAcylation was abolished in the PPV CP7-T/A mutant, in which seven threonines were mutated. PPV CP7-T/A infected Nicotiana clevelandii, Nicotiana benthamiana, and Prunus persica without noticeable defects. However, defects in infection of A. thaliana were readily apparent. In mixed infections of wild-type arabidopsis, the CP7-T/A mutant was outcompeted by wild-type virus. These results indicate that CP O-GlcNAcylation has a major role in the infection process. O-GlcNAc modification may have a role in virion assembly and/or stability as the CP of PPV CP7-T/A was more sensitive to protease digestion than that of the wild-type virus. Copyright © 2013 Elsevier Inc. All rights reserved.

Copper@carbon coaxial nanowires synthesized by hydrothermal carbonization process from electroplating wastewater and their use as an enzyme-free glucose sensor.

Science.gov (United States)

Zhao, Yuxin; He, Zhaoyang; Yan, Zifeng

2013-01-21

In the pursuit of electrocatalysts with great economic and ecological values for non-enzymatic glucose sensors, one-dimensional copper@carbon (Cu@C) core-shell coaxial nanowires (NWs) have been successfully prepared via a simple continuous flow wet-chemistry approach from electroplating wastewater. The as-obtained products were characterized by X-ray powder diffraction, scanning electron microscopy, transmission electron microscopy, selected area electron diffraction, energy dispersive X-ray spectroscopy and Raman spectroscopy. The electrocatalytic activity of the modified electrodes by Cu@C NWs towards glucose oxidation was investigated by cyclic voltammetry and chronoamperometry. It was found that the as-obtained Cu@C NWs showed good electrochemical properties and could be used as an electrochemical sensor for the detection of glucose molecules. Compared to the other electrodes including the bare Nafion/glassy carbon electrode (GCE) and several hot hybrid nanostructures modified GCE, a substantial decrease in the overvoltage of the glucose oxidation was observed at the Cu@C NWs electrodes with oxidation starting at ca. 0.20 V vs. Ag/AgCl (3 M KCl). At an applied potential of 0.65 V, Cu@C NWs electrodes had a high and reproducible sensitivity of 437.8 µA cm(-2) mM(-1) to glucose. Linear responses were obtained with a detection limit of 50 nM. More importantly, the proposed electrode also had good stability, high resistance against poisoning by chloride ion and commonly interfering species. These good analytical performances make Cu@C NWs promising for the future development of enzyme-free glucose sensors.

Measuring O-GlcNAc cleavage by OGA and cell lysates on a peptide microarray

NARCIS (Netherlands)

Sharif, Suhela; Shi, Jie; Bourakba, Mostafa; Ruijtenbeek, Rob; Pieters, Roland J.

2017-01-01

O-GlcNAcylation is a post-translational modification resulting from the addition of an N-acetylglucosamine moiety to the hydroxyl groups of serine and threonine residues of nuclear and cytoplasmic proteins. In addition, O-GlcNAcylated proteins can be phosphorylated, which suggests the possibility

Water-dispersible triethylenetetramine-functionalized graphene: Preparation, characterization and application as an amperometric glucose sensor

Energy Technology Data Exchange (ETDEWEB)

Ren, Qunxiang; Feng, Li; Fan, Ronghua; Ge, Xin; Sun, Yingying, E-mail: syyxiluda@hotmail.com

2016-11-01

The triethylenetetramine-functionalized graphene (TFGn) was prepared using graphene oxide (GO) and triethylenetetramine as raw materials through a one-step reaction under alkaline condition. The triethylenetetramine not only acted as cross-linker to combine GO, but also as reductant of GO. The TFGn was characterized by its ultraviolet spectrum (UV), Fourier transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), X-ray diffraction (XRD), Raman spectroscopy and Scanning electron microscopy (SEM). The results showed that triethylenetetramine was successfully grafted onto the surface of the GO through covalent bonding between amine and epoxy groups. The resultant TFGn was uniformly dispersed in water over several weeks, suggesting that the introduction of amino groups greatly increased the hydrophilicity of TFGn. The triethylenetetramine-functionalized graphene was then applied to fabricate glucose biosensors with IO{sub 4}{sup −} oxidized glucose oxidase (GOx) through layer-by-layer (LBL) self-assembly by the covalent bonding between the aldehyde groups of GOx and amino groups of TFGn. The gold electrodes modified with the (GOx/TFGn){sub n} multilayer films were studied by cyclic voltammetry (CV) and showed outstanding electrocatalytical response to the oxidation of glucose when ferrocenemethanol was used as an artificial redox mediator. The response increased with an increasing number of GOx/TFGn bilayers, indicating that the analytical performance, such as the sensitivity of the glucose biosensor, could be adjusted by tuning the number of deposited GOx/TFGn bilayers. The linear response range of the biosensor constructed with six bilayers of GOx/TFGn to the concentration of glucose can extend to at least 8 mM with a sensitivity of 19.9 μA mM{sup −1} cm{sup −2}. In addition, the sensor exhibited good stability due to the covalent interactions between the GOx and TFGn. - Highlights: • Water-dispersible triethylenetetramine

One-step simultaneous differential scanning calorimetry-FTIR microspectroscopy to quickly detect continuous pathways in the solid-state glucose/asparagine Maillard reaction.

Science.gov (United States)

Hwang, Deng-Fwu; Hsieh, Tzu-Feng; Lin, Shan-Yang

2013-01-01

The stepwise reaction pathway of the solid-state Maillard reaction between glucose (Glc) and asparagine (Asn) was investigated using simultaneous differential scanning calorimetry (DSC)-FTIR microspectroscopy. The color change and FTIR spectra of Glc-Asn physical mixtures (molar ratio = 1:1) preheated to different temperatures followed by cooling were also examined. The successive reaction products such as Schiff base intermediate, Amadori product, and decarboxylated Amadori product in the solid-state Glc-Asn Maillard reaction were first simultaneously evidenced by this unique DSC-FTIR microspectroscopy. The color changed from white to yellow-brown to dark brown, and appearance of new IR peaks confirmed the formation of Maillard reaction products. The present study clearly indicates that this unique DSC-FTIR technique not only accelerates but also detects precursors and products of the Maillard reaction in real time.

Phosphorylation coexists with O-GlcNAcylation in a plant virus protein and influences viral infection.

Science.gov (United States)

Martínez-Turiño, Sandra; Pérez, José De Jesús; Hervás, Marta; Navajas, Rosana; Ciordia, Sergio; Udeshi, Namrata D; Shabanowitz, Jeffrey; Hunt, Donald F; García, Juan Antonio

2018-06-01

Phosphorylation and O-GlcNAcylation are two widespread post-translational modifications (PTMs), often affecting the same eukaryotic target protein. Plum pox virus (PPV) is a member of the genus Potyvirus which infects a wide range of plant species. O-GlcNAcylation of the capsid protein (CP) of PPV has been studied extensively, and some evidence of CP phosphorylation has also been reported. Here, we use proteomics analyses to demonstrate that PPV CP is phosphorylated in vivo at the N-terminus and the beginning of the core region. In contrast with the 'yin-yang' mechanism that applies to some mammalian proteins, PPV CP phosphorylation affects residues different from those that are O-GlcNAcylated (serines Ser-25, Ser-81, Ser-101 and Ser-118). Our findings show that PPV CP can be concurrently phosphorylated and O-GlcNAcylated at nearby residues. However, an analysis using a differential proteomics strategy based on iTRAQ (isobaric tags for relative and absolute quantitation) showed a significant enhancement of phosphorylation at Ser-25 in virions recovered from O-GlcNAcylation-deficient plants, suggesting that crosstalk between O-GlcNAcylation and phosphorylation in PPV CP takes place. Although the preclusion of phosphorylation at the four identified phosphotarget sites only had a limited impact on viral infection, the mimicking of phosphorylation prevents PPV infection in Prunus persica and weakens infection in Nicotiana benthamiana and other herbaceous hosts, prompting the emergence of potentially compensatory second mutations. We postulate that the joint action of phosphorylation and O-GlcNAcylation in the N-proximal segment of CP allows a fine-tuning of protein stability, providing the amount of CP required in each step of viral infection. © 2017 BSPP AND JOHN WILEY & SONS LTD.

Senescence-Associated Changes in Proteome and O-GlcNAcylation Pattern in Human Peritoneal Mesothelial Cells

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Rebecca Herzog

2015-01-01

Full Text Available Introduction. Senescence of peritoneal mesothelial cells represents a biological program defined by arrested cell growth and altered cell secretory phenotype with potential impact in peritoneal dialysis. This study aims to characterize cellular senescence at the level of global protein expression profiles and modification of proteins with O-linked N-acetylglucosamine (O-GlcNAcylation. Methods. A comparative proteomics analysis between young and senescent human peritoneal mesothelial cells (HPMC was performed using two-dimensional gel electrophoresis. O-GlcNAc status was assessed by Western blot under normal conditions and after modulation with 6-diazo-5-oxo-L-norleucine (DON to decrease O-GlcNAcylation or O-(2-acetamido-2-deoxy-D-glucopyranosylidene amino N-phenyl carbamate (PUGNAc to increase O-GlcNAcylation. Results. Comparison of protein pattern of senescent and young HPMC revealed 29 differentially abundant protein spots, 11 of which were identified to be actin (cytoplasmic 1 and 2, cytokeratin-7, cofilin-2, transgelin-2, Hsp60, Hsc70, proteasome β-subunits (type-2 and type-3, nucleoside diphosphate kinase A, and cytosolic 5′(3′-deoxyribonucleotidase. Although the global level of O-GlcNAcylation was comparable, senescent cells were not sensitive to modulation by PUGNAc. Discussion. This study identified changes of the proteome and altered dynamics of O-GlcNAc regulation in senescent mesothelial cells. Whereas changes in cytoskeleton-associated proteins likely reflect altered cell morphology, changes in chaperoning and housekeeping proteins may have functional impact on cellular stress response in peritoneal dialysis.

Fabrication and Optimization of a Nanoporous Platinum Electrode and a Non-enzymatic Glucose Micro-sensor on Silicon

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Younghun Kim

2008-10-01

Full Text Available In this paper, optimal conditions for fabrication of nanoporous platinum (Pt were investigated in order to use it as a sensitive sensing electrode for silicon CMOS integrable non-enzymatic glucose micro-sensor applications. Applied charges, voltages, and temperatures were varied during the electroplating of Pt into the formed nonionic surfactant C16EO8 nano-scaled molds in order to fabricate nanoporous Pt electrodes with large surface roughness factor (RF, uniformity, and reproducibility. The fabricated nanoporous Pt electrodes were characterized using atomic force microscopy (AFM and electrochemical cyclic voltammograms. Optimal electroplating conditions were determined to be an applied charge of 35 mC/mm2, a voltage of -0.12 V, and a temperature of 25 °C, respectively. The optimized nanoporous Pt electrode had an electrochemical RF of 375 and excellent reproducibility. The optimized nanoporous Pt electrode was applied to fabricate non-enzymatic glucose micro-sensor with three electrode systems. The fabricated sensor had a size of 3 mm x 3 mm, air gap of 10 µm, working electrode (WE area of 4.4 mm2, and sensitivity of 37.5 µA•L/mmol•cm2. In addition, it showed large detection range from 0.05 to 30 mmolL-1 and stable recovery responsive to the step changes in glucose concentration.

Nanomaterial-based Electrochemical Sensors for the Detection of Glucose and Cholesterol

Science.gov (United States)

Ahmadalinezhad, Asieh

designed glucose biosensor exhibits a wide linear range, up to 18 mM glucose, as well as high sensitivity and selectivity. Glucose measurements of human serum using the developed biosensor showed excellent agreement with the data recorded by a commercial blood glucose monitoring assay. Finally, we fabricated an enzyme-free glucose sensor based on nanoporous palladium-cadmium (PdCd) networks. A hydrothermal method was applied in the synthesis of PdCd nanomaterials. The effect of the composition of the PdCd nanomaterials on the performance of the electrode was investigated by cyclic voltammetry (CV). Amperometric studies showed that the nanoporous PdCd electrode was responsive to the direct oxidation of glucose with high electrocatalytic activity. The sensitivity of the sensor for continuous glucose monitoring was 146.21 microAmM--1cm--2, with linearity up to 10 mM and a detection limit of 0.05 mM. In summary, the electrochemical biosensors proposed in my PhD study exhibited high sensitivity and selectivity for the continuous monitoring of analytes in the presence of common interference species. Our results have shown that the performance of the biosensors is significantly dependent on the dimensions and morphologies of nanostructured materials. The unique nanomaterials-based platforms proposed in this dissertation open the door to the design and fabrication of high-performance electrochemical biosensors for medical diagnostics.

Theoretical pKa prediction of the α-phosphate moiety of uridine 5‧-diphosphate-GlcNAc

Science.gov (United States)

Vipperla, Bhavaniprasad; Griffiths, Thomas M.; Wang, Xingyong; Yu, Haibo

2017-01-01

The pKa value of the α-phosphate moiety of uridine 5‧-diphosphate-GlcNAc (UDP-GlcNAc) has been successfully calculated using density functional theory methods in conjunction with the Polarizable Continuum Models. Theoretical methods were benchmarked over a dataset comprising of alkyl phosphates. B3LYP/6-31+G(d,p) calculations using SMD solvation model provide excellent agreement with the experimental data. The predicted pKa for UDP-GlcNAc is consistent with most recent NMR studies but much higher than what it has long been thought to be. The importance of this study is evident that the predicted pKa for UDP-GlcNAc supports its potential role as a catalytic base in the substrate-assisted biocatalysis.

A Novel Microdialysis Glucose Sensor System Based on Co-immobilizing on AU Micro-Electrode by SOL-GEL Technique

National Research Council Canada - National Science Library

Yu, Ping

2001-01-01

.... The sensor is based on co_immobilizing glucose oxidase (COD) with the catalase by sol-gel technique on the surface of the silicon bases with two pattern of An microelectrodes. A graduated ("sandwich...

Augmented O-GlcNAcylation of AMP-activated kinase promotes the proliferation of LoVo cells, a colon cancer cell line.

Science.gov (United States)

Ishimura, Emi; Nakagawa, Takatoshi; Moriwaki, Kazumasa; Hirano, Seiichi; Matsumori, Yoshinobu; Asahi, Michio

2017-12-01

Increasing incidence of various cancers has been reported in diabetic patients. O-linked N-acetylglucosamine (O-GlcNAc) modification of proteins at serine/threonine residues (O-GlcNAcylation) is an essential post-translational modification that is upregulated in diabetic patients and has been implicated in tumor growth. However, the mechanisms by which O-GlcNAcylation promotes tumor growth remain unclear. Given that AMP-activated kinase (AMPK) has been thought to play important roles in suppressing tumor growth, we evaluated the involvement of AMPK O-GlcNAcylation on the growth of LoVo cells, a human colon cancer cell line. Results revealed that treatment with Thiamet G (TMG), an inhibitor of O-GlcNAc hydrolase, increased both anchorage-dependent and -independent growth of the cells. O-GlcNAc transferase overexpression also increased the growth. These treatments increased AMPK O-GlcNAcylation in a dose-dependent manner, which led to reduced AMPK phosphorylation and mTOR activation. Chemical inhibition or activation of AMPK led to increased or decreased growth, respectively, which was consistent with the data with genetic inhibition of AMPK. In addition, TMG-mediated acceleration of tumor growth was abolished by both chemical and genetic inhibition of AMPK. To examine the effects of AMPK O-GlcNAcylation in vivo, the LoVo cells were s.c. transplanted onto the backs of BALB/c-nu/nu mice. Injection of TMG promoted the growth and enhanced O-GlcNAcylation of the tumors of the mice. Consistent with in vitro data, AMPK O-GlcNAcylation was increased, which reduced AMPK phosphorylation and resulted in activation of mTOR. Collectively, the higher colon cancer risk of diabetic patients could be due to O-GlcNAcylation-mediated AMPK inactivation and subsequent activation of mTOR. © 2017 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.

O-linked-N-acetylglucosamine modification of mammalian Notch receptors by an atypical O-GlcNAc transferase Eogt1

Energy Technology Data Exchange (ETDEWEB)

Sakaidani, Yuta [Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065 (Japan); Ichiyanagi, Naoki [Department of Applied Molecular Biosciences, Nagoya University Graduate School of Bioagricultural Sciences, Furo-cho, Chikusa-ku, Nagoya 464-8601 (Japan); Saito, Chika; Nomura, Tomoko [Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065 (Japan); Ito, Makiko [Department of Applied Molecular Biosciences, Nagoya University Graduate School of Bioagricultural Sciences, Furo-cho, Chikusa-ku, Nagoya 464-8601 (Japan); Nishio, Yosuke [Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065 (Japan); Nadano, Daita; Matsuda, Tsukasa [Department of Applied Molecular Biosciences, Nagoya University Graduate School of Bioagricultural Sciences, Furo-cho, Chikusa-ku, Nagoya 464-8601 (Japan); Furukawa, Koichi [Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065 (Japan); Okajima, Tetsuya, E-mail: tokajima@med.nagoya-u.ac.jp [Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065 (Japan); Department of Applied Molecular Biosciences, Nagoya University Graduate School of Bioagricultural Sciences, Furo-cho, Chikusa-ku, Nagoya 464-8601 (Japan)

2012-03-02

Highlights: Black-Right-Pointing-Pointer We characterized A130022J15Rik (Eogt1)-a mouse gene homologous to Drosophila Eogt. Black-Right-Pointing-Pointer Eogt1 encodes EGF domain O-GlcNAc transferase. Black-Right-Pointing-Pointer Expression of Eogt1 in Drosophila rescued the cell-adhesion defect in the Eogt mutant. Black-Right-Pointing-Pointer O-GlcNAcylation reaction in the secretory pathway is conserved through evolution. -- Abstract: O-linked-{beta}-N-acetylglucosamine (O-GlcNAc) modification is a unique cytoplasmic and nuclear protein modification that is common in nearly all eukaryotes, including filamentous fungi, plants, and animals. We had recently reported that epidermal growth factor (EGF) repeats of Notch and Dumpy are O-GlcNAcylated by an atypical O-GlcNAc transferase, EOGT, in Drosophila. However, no study has yet shown whether O-GlcNAcylation of extracellular proteins is limited to insects such as Drosophila or whether it occurs in other organisms, including mammals. Here, we report the characterization of A130022J15Rik, a mouse gene homolog of Drosophila Eogt (Eogt 1). Enzymatic analysis revealed that Eogt1 has a substrate specificity similar to that of Drosophila EOGT, wherein the Thr residue located between the fifth and sixth conserved cysteines of the folded EGF-like domains is modified. This observation is supported by the fact that the expression of Eogt1 in Drosophila rescued the cell-adhesion defect caused by Eogt downregulation. In HEK293T cells, Eogt1 expression promoted modification of Notch1 EGF repeats by O-GlcNAc, which was further modified, at least in part, by galactose to generate a novel O-linked-N-acetyllactosamine structure. These results suggest that Eogt1 encodes EGF domain O-GlcNAc transferase and that O-GlcNAcylation reaction in the secretory pathway is a fundamental biochemical process conserved through evolution.

O-linked-N-acetylglucosamine modification of mammalian Notch receptors by an atypical O-GlcNAc transferase Eogt1

International Nuclear Information System (INIS)

Sakaidani, Yuta; Ichiyanagi, Naoki; Saito, Chika; Nomura, Tomoko; Ito, Makiko; Nishio, Yosuke; Nadano, Daita; Matsuda, Tsukasa; Furukawa, Koichi; Okajima, Tetsuya

2012-01-01

Highlights: ► We characterized A130022J15Rik (Eogt1)—a mouse gene homologous to Drosophila Eogt. ► Eogt1 encodes EGF domain O-GlcNAc transferase. ► Expression of Eogt1 in Drosophila rescued the cell-adhesion defect in the Eogt mutant. ► O-GlcNAcylation reaction in the secretory pathway is conserved through evolution. -- Abstract: O-linked-β-N-acetylglucosamine (O-GlcNAc) modification is a unique cytoplasmic and nuclear protein modification that is common in nearly all eukaryotes, including filamentous fungi, plants, and animals. We had recently reported that epidermal growth factor (EGF) repeats of Notch and Dumpy are O-GlcNAcylated by an atypical O-GlcNAc transferase, EOGT, in Drosophila. However, no study has yet shown whether O-GlcNAcylation of extracellular proteins is limited to insects such as Drosophila or whether it occurs in other organisms, including mammals. Here, we report the characterization of A130022J15Rik, a mouse gene homolog of Drosophila Eogt (Eogt 1). Enzymatic analysis revealed that Eogt1 has a substrate specificity similar to that of Drosophila EOGT, wherein the Thr residue located between the fifth and sixth conserved cysteines of the folded EGF-like domains is modified. This observation is supported by the fact that the expression of Eogt1 in Drosophila rescued the cell-adhesion defect caused by Eogt downregulation. In HEK293T cells, Eogt1 expression promoted modification of Notch1 EGF repeats by O-GlcNAc, which was further modified, at least in part, by galactose to generate a novel O-linked-N-acetyllactosamine structure. These results suggest that Eogt1 encodes EGF domain O-GlcNAc transferase and that O-GlcNAcylation reaction in the secretory pathway is a fundamental biochemical process conserved through evolution.

The ethylene glycol template assisted hydrothermal synthesis of Co{sub 3}O{sub 4} nanowires; structural characterization and their application as glucose non-enzymatic sensor

Energy Technology Data Exchange (ETDEWEB)

Khun, K., E-mail: kimleang.khun@liu.se [Department of Science and Technology, Linköping University, SE-60174 Norrköping (Sweden); Ibupoto, Z.H. [Dr M.A. Kazi Institute of Chemistry, University of Sindh Jamshoro, Sindh Jamshoro (Pakistan); Liu, X. [Department of Physics, Chemistry and Biology, Linköping University, 58183 Linköping (Sweden); Beni, V. [Biosensors and Biolelectronics Centre, Department of Physics, Chemistry and Biology, Linköping University, 58183 Linköping (Sweden); Willander, M. [Department of Science and Technology, Linköping University, SE-60174 Norrköping (Sweden)

2015-04-15

Highlights: • Ethylene glycol assisted Co{sub 3}O{sub 4} nanowires were synthesized by hydrothermal method. • The grown Co{sub 3}O{sub 4} nanowires were used for sensitive non-enzymatic glucose sensor. • The proposed glucose sensor shows a wide linear range with fast response. • The Co{sub 3}O{sub 4} modified electrode is a highly specific enzyme-less glucose sensor. - Abstract: In the work reported herein the ethylene glycol template assisted hydrothermal synthesis, onto Au substrate, of thin and highly dense cobalt oxide (Co{sub 3}O{sub 4}) nanowires and their characterization and their application for non-enzymatic glucose sensing are reported. The structure and composition of Co{sub 3}O{sub 4} nanowires have been fully characterized using scanning electron microscopy, X-ray diffraction, high resolution transmission electron microscopy and X-ray photoelectron spectroscopy. The synthesized Co{sub 3}O{sub 4} nanowires resulted to have high purity and showed diameter of approximately 10 nm. The prepared Co{sub 3}O{sub 4} nanowires coated gold electrodes were applied to the non-enzymatic detection of glucose. The developed sensor showed high sensitivity (4.58 × 10{sup 1} μA mM{sup −1} cm{sup −2}), a wide linear range of concentration (1.00 × 10{sup −4}–1.2 × 10{sup 1} mM) and a detection limit of 2.65 × 10{sup −5} mM. The developed glucose sensor has also shown to be very stable and selective over interferents such as uric acid and ascorbic acid. Furthermore, the proposed fabrication process was shown to be highly reproducible response (over nine electrodes)

Identification of GIG1, a GlcNAc-induced gene in Candida albicans needed for normal sensitivity to the chitin synthase inhibitor nikkomycin Z.

Science.gov (United States)

Gunasekera, Angelo; Alvarez, Francisco J; Douglas, Lois M; Wang, Hong X; Rosebrock, Adam P; Konopka, James B

2010-10-01

The amino sugar N-acetylglucosamine (GlcNAc) is known to be an important structural component of cells from bacteria to humans, but its roles in cell signaling are less well understood. GlcNAc induces two pathways in the human fungal pathogen Candida albicans. One activates cyclic AMP (cAMP) signaling, which stimulates the formation of hyphal cells and the expression of virulence genes, and the other pathway induces genes needed to catabolize GlcNAc. Microarray analysis of gene expression was carried out under four different conditions in order to characterize the transcriptional changes induced by GlcNAc. The most highly induced genes include those that encode a GlcNAc transporter (NGT1) and the GlcNAc catabolic enzymes (HXK1, DAC1, and NAG1). GlcNAc also activated most of the genes whose expression is increased when cells are triggered with other stimuli to form hyphae. Surprisingly, GlcNAc also induced a subset of genes that are regulated by galactose (GAL1, GAL7, and GAL10), which may be due to cross talk between signaling pathways. A novel GlcNAc-induced gene, GIG1, which is not essential for GlcNAc catabolism or the induction of hyphae, was identified. However, a Gig1-green fluorescent protein (GFP) fusion protein was specifically induced by GlcNAc, and not by other sugars. Gig1-GFP localized to the cytoplasm, where GlcNAc metabolism occurs. Significantly, a gig1Δ mutant displayed increased resistance to nikkomycin Z, which inhibits chitin synthase from converting UDP-GlcNAc into cell wall chitin. Gig1 is highly conserved in fungi, especially those that contain GlcNAc catabolic genes. These results implicate Gig1 in GlcNAc metabolism.

Chemical exchange-sensitive spin-lock MRI of glucose analog 3-O-methyl-d-glucose in normal and ischemic brain.

Science.gov (United States)

Jin, Tao; Mehrens, Hunter; Wang, Ping; Kim, Seong-Gi

2018-05-01

Glucose transport is important for understanding brain glucose metabolism. We studied glucose transport with a presumably non-toxic and non-metabolizable glucose analog, 3-O-methyl-d-glucose, using a chemical exchange-sensitive spin-lock MRI technique at 9.4 Tesla. 3-O-methyl-d-glucose showed comparable chemical exchange properties with d-glucose and 2-deoxy-d-glucose in phantoms, and higher and lower chemical exchange-sensitive spin-lock sensitivity than Glc and 2-deoxy-d-glucose in in vivo experiments, respectively. The changes of the spin-lattice relaxation rate in the rotating frame (Δ R 1 ρ) in normal rat brain peaked at ∼15 min after the intravenous injection of 1 g/kg 3-O-methyl-d-glucose and almost maintained a plateau for >1 h. Doses up to 4 g/kg 3-O-methyl-d-glucose were linearly correlated with Δ R 1 ρ. In rats with focal ischemic stroke, chemical exchange-sensitive spin-lock with 3-O-methyl-d-glucose injection at 1 h after stroke onset showed reduced Δ R 1 ρ in the ischemic core but higher Δ R 1 ρ in the peri-core region compared to normal tissue, which progressed into the ischemic core at 3 h after stroke onset. This suggests that the hyper-chemical exchange-sensitive spin-lock region observed at 1 h is the ischemic penumbra at-risk of infarct. In summary, 3-O-methyl-d-glucose-chemical exchange-sensitive spin-lock can be a sensitive MRI technique to probe the glucose transport in normal and ischemic brains.

[Apoptosis of human lung carcinoma cell line GLC-82 induced by high power electromagnetic pulse].

Science.gov (United States)

Cao, Xiao-zhe; Zhao, Mei-lan; Wang, De-wen; Dong, Bo

2002-09-01

Electromagnetic pulse (EMP) could be used for sterilization of food and the efficiency is higher than 2450 MHz continuous microwave done. This study was designed to evaluate the effect of electromagnetic pulse (EMP) on apoptosis of human lung carcinoma cell line GLC-82, so that to explore and develop therapeutic means for cancer. The injury changes in GLC-82 cells after irradiated with EMP (electric field intensity was 60 kV/m, 5 pulses/2 min) were analyzed by cytometry, MTT chronometry, and flow cytometry. The immunohistochemical SP staining was used to determine the expressions of bcl-2 protein and p53 protein. The stained positive cells were analyzed by CMIAS-II image analysis system at a magnification 400. All data were analyzed by SPSS8.0 software. EMP could obviously inhibited proliferation and activity of lung carcinoma cell line GLC-82. The absorbance value (A570) of MTT decreased immediately, at 0 h, 1 h, and 6 h after the GLC-82 cells irradiated by EMP as compared with control group. The highest apoptosis rate was found to reach 13.38% by flow cytometry at 6 h after EMP irradiation. Down-regulation of bcl-2 expression and up-regulation of p53 expression were induced by EMP. EMP promotes apoptosis of GLC-82 cells. At same time, EMP can down-regulate bcl-2 expression and up-regulate p53 expression in GLC-82 cells. The bcl-2 and the p53 protein may involve the apoptotic process.

Glucose Elevates NITRATE TRANSPORTER2.1 Protein Levels and Nitrate Transport Activity Independently of Its HEXOKINASE1-Mediated Stimulation of NITRATE TRANSPORTER2.1 Expression1[W][OPEN

Science.gov (United States)

de Jong, Femke; Thodey, Kate; Lejay, Laurence V.; Bevan, Michael W.

2014-01-01

Mineral nutrient uptake and assimilation is closely coordinated with the production of photosynthate to supply nutrients for growth. In Arabidopsis (Arabidopsis thaliana), nitrate uptake from the soil is mediated by genes encoding high- and low-affinity transporters that are transcriptionally regulated by both nitrate and photosynthate availability. In this study, we have studied the interactions of nitrate and glucose (Glc) on gene expression, nitrate transport, and growth using glucose-insensitive2-1 (gin2-1), which is defective in sugar responses. We confirm and extend previous work by showing that HEXOKINASE1-mediated oxidative pentose phosphate pathway (OPPP) metabolism is required for Glc-mediated NITRATE TRANSPORTER2.1 (NRT2.1) expression. Treatment with pyruvate and shikimate, two products derived from intermediates of the OPPP that are destined for amino acid production, restores wild-type levels of NRT2.1 expression, suggesting that metabolites derived from OPPP metabolism can, together with Glc, directly stimulate high levels of NRT2.1 expression. Nitrate-mediated NRT2.1 expression is not influenced by gin2-1, showing that Glc does not influence NRT2.1 expression through nitrate-mediated mechanisms. We also show that Glc stimulates NRT2.1 protein levels and transport activity independently of its HEXOKINASE1-mediated stimulation of NRT2.1 expression, demonstrating another possible posttranscriptional mechanism influencing nitrate uptake. In gin2-1 plants, nitrate-responsive biomass growth was strongly reduced, showing that the supply of OPPP metabolites is essential for assimilating nitrate for growth. PMID:24272701

A label-free fiber-optic Turbidity Affinity Sensor (TAS) for continuous glucose monitoring.

Science.gov (United States)

Dutt-Ballerstadt, Ralph; Evans, Colton; Pillai, Arun P; Gowda, Ashok

2014-11-15

In this paper, we describe the concept of a novel implantable fiber-optic Turbidity Affinity Sensor (TAS) and report on the findings of its in-vitro performance for continuous glucose monitoring. The sensing mechanism of the TAS is based on glucose-specific changes in light scattering (turbidity) of a hydrogel suspension consisting of small particles made of crosslinked dextran (Sephadex G100), and a glucose- and mannose-specific binding protein - Concanavalin A (ConA). The binding of ConA to Sephadex particles results in a significant turbidity increase that is much greater than the turbidity contribution by the individual components. The turbidity of the TAS was measured by determining the intensity of light passing through the suspension enclosed within a small semi-permeable hollow fiber (OD: 220 μm, membrane thickness: 20 μm, molecular weight cut-off: 10 kDa) using fiber optics. The intensity of measured light of the TAS was proportional to the glucose concentration over the concentration range from 50mg/dL to 400mg/dL in PBS and whole blood at 37°C (R>0.96). The response time was approximately 4 min. The stability of the glucose response of the TAS decreased only slightly (by 20%) over an 8-day study period at 37°C. In conclusion, this study demonstrated proof-of-concept of the TAS for interstitial glucose monitoring. Due to the large signal amplitude of the turbidity change, and the lack of need for wavelength-specific emission and excitation filters, a very small, robust and compact TAS device with an extremely short optical pathlength could be feasibly designed and implemented for in-vivo glucose monitoring in people with diabetes. Copyright © 2014 Elsevier B.V. All rights reserved.

Accuracy and precision of flash glucose monitoring sensors inserted into the abdomen and upper thigh compared with the upper arm.

Science.gov (United States)

Charleer, Sara; Mathieu, Chantal; Nobels, Frank; Gillard, Pieter

2018-06-01

Nowadays, most Belgian patients with type 1 diabetes use flash glucose monitoring (FreeStyle Libre [FSL]; Abbott Diabetes Care, Alameda, California) to check their glucose values, but some patients find the sensor on the upper arm too visible. The aim of the present study was to compare the accuracy and precision of FSL sensors when placed on different sites. A total of 23 adults with type 1 diabetes used three FSL sensors simultaneously for 14 days on the upper arm, abdomen and upper thigh. FSL measurements were compared with capillary blood glucose (BG) measurements obtained with a built-in FSL BG meter. The aggregated mean absolute relative difference was 11.8 ± 12.0%, 18.5 ± 18.4% and 12.3 ± 13.8% for the arm, abdomen (P = .002 vs arm) and thigh (P = .5 vs arm), respectively. Results of Clarke error grid analysis for the arm and thigh were similar (zone A: 84.9% vs 84.5%; P = .6), while less accuracy was seen for the abdomen (zone A: 69.4%; P = .01). Apart from the first day, the accuracy of FSL sensors on the arm and thigh was more stable across the 14-day wear duration than accuracy of sensors on the abdomen, which deteriorated mainly during week 2 (P < .0005). The aggregated precision absolute relative difference was markedly lower for the arm/thigh (10.9 ± 11.9%) compared with the arm/abdomen (20.9 ± 22.8%; P = .002). Our results indicate that the accuracy and precision of FSL sensors placed on the upper thigh are similar to the upper arm, whereas the abdomen performed unacceptably poorly. © 2018 John Wiley & Sons Ltd.

Cell cycle-dependent O-GlcNAc modification of tobacco histones and their interaction with the tobacco lectin.

Science.gov (United States)

Delporte, Annelies; De Zaeytijd, Jeroen; De Storme, Nico; Azmi, Abdelkrim; Geelen, Danny; Smagghe, Guy; Guisez, Yves; Van Damme, Els J M

2014-10-01

The Nicotiana tabacum agglutinin or Nictaba is a nucleocytoplasmic lectin that is expressed in tobacco after the plants have been exposed to jasmonate treatment or insect herbivory. Nictaba specifically recognizes GlcNAc residues. Recently, it was shown that Nictaba is interacting in vitro with the core histone proteins from calf thymus. Assuming that plant histones - similar to their animal counterparts - undergo O-GlcNAcylation, this interaction presumably occurs through binding of the lectin to the O-GlcNAc modification present on the histones. Hereupon, the question was raised whether this modification also occurs in plants and if it is cell cycle dependent. To this end, histones were purified from tobacco BY-2 suspension cells and the presence of O-GlcNAc modifications was checked. Concomitantly, O-GlcNAcylation of histone proteins was studied. Our data show that similar to animal histones plant histones are modified by O-GlcNAc in a cell cycle-dependent fashion. In addition, the interaction between Nictaba and tobacco histones was confirmed using lectin chromatography and far Western blot analysis. Collectively these findings suggest that Nictaba can act as a modulator of gene transcription through its interaction with core histones. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

Polymer optical fiber compound parabolic concentrator tip for enhanced coupling efficiency for fluorescence based glucose sensors

DEFF Research Database (Denmark)

Hassan, Hafeez Ul; Nielsen, Kristian; Aasmul, Søren

2015-01-01

We demonstrate that the light excitation and capturing efficiency of fluorescence based fiber-optical sensors can be significantly increased by using a CPC (Compound Parabolic Concentrator) tip instead of the standard plane-cut tip. We use Zemax modelling to find the optimum CPC tip profile...... and fiber length of a polymer optical fiber diabetes sensor for continuous monitoring of glucose levels. We experimentally verify the improved performance of the CPC tipped sensor and the predicted production tolerances. Due to physical size requirements when the sensor has to be inserted into the body...... a non-optimal fiber length of 35 mm is chosen. For this length an average improvement in efficiency of a factor of 1.7 is experimentally demonstrated and critically compared to the predicted ideal factor of 3 in terms of parameters that should be improved through production optimization....

Polymer optical fiber compound parabolic concentrator tip for enhanced coupling efficiency for fluorescence based glucose sensors.

Science.gov (United States)

Hassan, Hafeez Ul; Nielsen, Kristian; Aasmul, Soren; Bang, Ole

2015-12-01

We demonstrate that the light excitation and capturing efficiency of fluorescence based fiber-optical sensors can be significantly increased by using a CPC (Compound Parabolic Concentrator) tip instead of the standard plane-cut tip. We use Zemax modelling to find the optimum CPC tip profile and fiber length of a polymer optical fiber diabetes sensor for continuous monitoring of glucose levels. We experimentally verify the improved performance of the CPC tipped sensor and the predicted production tolerances. Due to physical size requirements when the sensor has to be inserted into the body a non-optimal fiber length of 35 mm is chosen. For this length an average improvement in efficiency of a factor of 1.7 is experimentally demonstrated and critically compared to the predicted ideal factor of 3 in terms of parameters that should be improved through production optimization.

A glucose oxidase-coupled DNAzyme sensor for glucose detection in tears and saliva.

Science.gov (United States)

Liu, Chengcheng; Sheng, Yongjie; Sun, Yanhong; Feng, Junkui; Wang, Shijin; Zhang, Jin; Xu, Jiacui; Jiang, Dazhi

2015-08-15

Biosensors have been widely investigated and utilized in a variety of fields ranging from environmental monitoring to clinical diagnostics. Glucose biosensors have triggered great interest and have been widely exploited since glucose determination is essential for diabetes diagnosis. In here, we designed a novel dual-enzyme biosensor composed of glucose oxidase (GOx) and pistol-like DNAzyme (PLDz) to detect glucose levels in tears and saliva. First, GOx, as a molecular recognition element, catalyzes the oxidation of glucose forming H2O2; then PLDz recognizes the produced H2O2 as a secondary signal and performs a self-cleavage reaction promoted by Mn(2+), Co(2+) and Cu(2+). Thus, detection of glucose could be realized by monitoring the cleavage rate of PLDz. The slope of the cleavage rate of PLDz versus glucose concentration curve was fitted with a Double Boltzmann equation, with a range of glucose from 100 nM to 10mM and a detection limit of 5 μM. We further applied the GOx-PLDz 1.0 biosensor for glucose detection in tears and saliva, glucose levels in which are 720±81 μM and 405±56 μM respectively. Therefore, the GOx-PLDz 1.0 biosensor is able to determine glucose levels in tears and saliva as a noninvasive glucose biosensor, which is important for diabetic patients with frequent/continuous glucose monitoring requirements. In addition, induction of DNAzyme provides a new approach in the development of glucose biosensors. Copyright © 2015 Elsevier B.V. All rights reserved.

Pseudo-bi-enzyme glucose sensor: ZnS hollow spheres and glucose oxidase concerted catalysis glucose.

Science.gov (United States)

Shuai, Ying; Liu, Changhua; Wang, Jia; Cui, Xiaoyan; Nie, Ling

2013-06-07

This work creatively uses peroxidase-like ZnS hollow spheres (ZnS HSs) to cooperate with glucose oxidase (GOx) for glucose determinations. This approach is that the ZnS HSs electrocatalytically oxidate the enzymatically generated H2O2 to O2, and then the O2 circularly participates in the previous glucose oxidation by glucose oxidase. Au nanoparticles (AuNPs) and carbon nanotubes (CNTs) are used as electron transfer and enzyme immobilization matrices, respectively. The biosensor of glucose oxidase-carbon nanotubes-Au nanoparticles-ZnS hollow spheres-gold electrode (GOx-CNT-AuNPs-ZnS HSs-GE) exhibits a rapid response, a low detection limit (10 μM), a wide linear range (20 μM to 7 mM) as well as good anti-interference, long-term longevity and reproducibility.

[Tetra-saccharide glucose as a diagnostic biomarker for Pompe disease: a study with 35 patients].

Science.gov (United States)

Bobillo Lobato, Joaquín; Durán Parejo, Pilar; Tejero Díez, Pedro; Jiménez Jiménez, Luis M

2013-08-04

Pompe disease is a disorder originating from an acid alpha-glycosidase (AAG) enzyme deficiency. This disease produces an accumulation of lysosomal glycogen in different tissues, whereby the skeletal and heart muscles are especially involved. The established diagnosis is achieved through the identification of the AAG deficiency. There are also other secondary diagnostic biomarkers, such as tetra-saccharide glucose (Glc4), which shows high levels in the urine of these patients. In this study it is highlighted the usefulness of Glc4 as a diagnostic biomarker for Pompe disease in its different forms of presentation, using a high-performance liquid chromatography with ultraviolet detection (HPLC/UV) adapted to the study. A total of 75 individuals have been analyzed: 40 healthy controls and 35 patients diagnosed with Pompe disease. Twenty-four hour samples of urine were collected from all of the patients and their Glc4 levels were determined by means of HPLC/UV. The evaluation of the urinary Glc4 shows a high discrimination ability between healthy/sick individuals. In addition, the results obtained have allowed to establish the most appropriate level of decision or cut-off point for the identification of sick people. Glc4 urinary levels are found to be high in patients suffering from Pompe disease and even though increased levels are also found in other conditions, the existence of a AAG deficiency together with a compatible clinical symptoms, prove very helpful for a correct diagnosis of this serious disease. Copyright © 2012 Elsevier España, S.L. All rights reserved.

Identification of GIG1, a GlcNAc-Induced Gene in Candida albicans Needed for Normal Sensitivity to the Chitin Synthase Inhibitor Nikkomycin Z▿§

Science.gov (United States)

Gunasekera, Angelo; Alvarez, Francisco J.; Douglas, Lois M.; Wang, Hong X.; Rosebrock, Adam P.; Konopka, James B.

2010-01-01

The amino sugar N-acetylglucosamine (GlcNAc) is known to be an important structural component of cells from bacteria to humans, but its roles in cell signaling are less well understood. GlcNAc induces two pathways in the human fungal pathogen Candida albicans. One activates cyclic AMP (cAMP) signaling, which stimulates the formation of hyphal cells and the expression of virulence genes, and the other pathway induces genes needed to catabolize GlcNAc. Microarray analysis of gene expression was carried out under four different conditions in order to characterize the transcriptional changes induced by GlcNAc. The most highly induced genes include those that encode a GlcNAc transporter (NGT1) and the GlcNAc catabolic enzymes (HXK1, DAC1, and NAG1). GlcNAc also activated most of the genes whose expression is increased when cells are triggered with other stimuli to form hyphae. Surprisingly, GlcNAc also induced a subset of genes that are regulated by galactose (GAL1, GAL7, and GAL10), which may be due to cross talk between signaling pathways. A novel GlcNAc-induced gene, GIG1, which is not essential for GlcNAc catabolism or the induction of hyphae, was identified. However, a Gig1-green fluorescent protein (GFP) fusion protein was specifically induced by GlcNAc, and not by other sugars. Gig1-GFP localized to the cytoplasm, where GlcNAc metabolism occurs. Significantly, a gig1Δ mutant displayed increased resistance to nikkomycin Z, which inhibits chitin synthase from converting UDP-GlcNAc into cell wall chitin. Gig1 is highly conserved in fungi, especially those that contain GlcNAc catabolic genes. These results implicate Gig1 in GlcNAc metabolism. PMID:20675577

Determination of a glucose-containing tetrasaccharide in urine by radioimmunoassay

International Nuclear Information System (INIS)

Zopf, D.A.; Levinson, R.E.; Lundblad, A.

1982-01-01

A radioimmunoassay is described that allows rapid determination of a urinary oligosaccharide - Glcα1-6Glcα1-4Glcα1-4Glc [(Glc) 4 ] - at concentrations >2pmol/μl. Antibodies produced in rabbits immunized with the phenethylamine derivative of (Glc) 4 coupled to keyhole limpet hemocyanin bind tritiated (Glc) 4 -alditol. Studies comparing the activities of (Glc) 4 and several of its derivatives and analogues as inhibitors of binding of tritiated (Glc) 4 -alditol show that (Glc) 4 is detected with maximum specificity and sensitivity only after reduction to (Glc) 4 -alditol. Quantitation of (Glc) 4 in urine samples reduced with sodium borohydride gives excellent agreement with a previously used but more laborious method that employs gas-liquid chromatography-mass spectrometry (GLC/MS) (correlation coefficient = 0.98). Studies of the excretion rate of (Glc) 4 in urine of women during pregnancy using the radioimmunoassay method confirm and extend previous results using the GLC/MS method. (Auth.)

Suppressive role of OGT-mediated O-GlcNAcylation of BAP1 in retinoic acid signaling.

Science.gov (United States)

Moon, Seungtae; Lee, Yong-Kyu; Lee, Sang-Wang; Um, Soo-Jong

2017-10-07

BRCA1-associated protein 1 (BAP1) has been implicated in diverse biological functions, including tumor suppression. However, its regulation via glycosylation and its role in embryonic stem (ES) cells are poorly defined. BAP1 was recently reported to interact with O-linked N-acetylglucosamine (O-GlcNAc) transferase (OGT). Here, we confirmed the physical interaction and investigated its functional significance. The O-GlcNAcylation of BAP1, which requires OGT, was examined in vivo and in vitro, and was proven using alloxan, an OGT inhibitor. OGT promoted the BAP1-induced repression of retinoic acid (RA)-induced RA receptor (RAR) activation. The repressive activity of BAP1 was relieved by alloxan but exacerbated by PUGNAc, an O-GlcNAcase (OGA) inhibitor. Finally, we addressed the role of O-GlcNAcylation in the RA-induced differentiation of murine ES cells. Alkaline phosphatase staining revealed the cooperation of RA and alloxan for impairing the pluripotency of ES cells. This cooperation was also observed by measuring the size of embryonic bodies and the expression of Sox2, a pluripotency marker. Overall, our data suggest that OGT-mediated O-GlcNAcylation of BAP1 prefers the maintenance of pluripotency, whereas its inhibition facilitates RA-induced differentiation in ES cells. Copyright © 2017 Elsevier Inc. All rights reserved.

Determination of a glucose-containing tetrasaccharide in urine by radioimmunoassay

Energy Technology Data Exchange (ETDEWEB)

Zopf, D.A.; Levinson, R.E. (National Cancer Inst., Bethesda, MD (USA)); Lundblad, A. (University Hospital, Lund (Sweden). Dept. of Clinical Chemistry)

1982-01-15

A radioimmunoassay is described that allows rapid determination of a urinary oligosaccharide - Glc..cap alpha..1-6Glc..cap alpha..1-4Glc..cap alpha..1-4Glc ((Glc)/sub 4/) - at concentrations >2pmol/..mu..l. Antibodies produced in rabbits immunized with the phenethylamine derivative of (Glc)/sub 4/ coupled to keyhole limpet hemocyanin bind tritiated (Glc)/sub 4/-alditol. Studies comparing the activities of (Glc)/sub 4/ and several of its derivatives and analogues as inhibitors of binding of tritiated (Glc)/sub 4/-alditol show that (Glc)/sub 4/ is detected with maximum specificity and sensitivity only after reduction to (Glc)/sub 4/-alditol. Quantitation of (Glc)/sub 4/ in urine samples reduced with sodium borohydride gives excellent agreement with a previously used but more laborious method that employs gas-liquid chromatography-mass spectrometry (GLC/MS) (correlation coefficient = 0.98). Studies of the excretion rate of (Glc)/sub 4/ in urine of women during pregnancy using the radioimmunoassay method confirm and extend previous results using the GLC/MS method.

From GLC to double-null coordinates and illustration with static black holes

Energy Technology Data Exchange (ETDEWEB)

Nugier, Fabien, E-mail: fnugier@ntu.edu.tw [Leung Center for Cosmology and Particle Astrophysics, National Taiwan University, Taipei, 10617 Taiwan (China)

2016-09-01

We present a system of coordinates deriving directly from the so-called Geodesic Light-Cone (GLC) coordinates and made of two null scalars intersecting on a 2-dimensional sphere parameterized by two constant angles along geodesics. These coordinates are shown to be equivalent to the well-known double-null coordinates. As GLC, they present interesting properties for cosmology and astrophysics. We discuss this latter topic for static black holes, showing simple descriptions for the metric or particles and photons trajectories. We also briefly comment on the time of flight of ultra-relativistic particles.

Time-domain fiber loop ringdown sensor and sensor network

Science.gov (United States)

Kaya, Malik

Optical fibers have been mostly used in fiber optic communications, imaging optics, sensing technology, etc. Fiber optic sensors have gained increasing attention for scientific and structural health monitoring (SHM) applications. In this study, fiber loop ringdown (FLRD) sensors were fabricated for scientific, SHM, and sensor networking applications. FLRD biosensors were fabricated for both bulk refractive index (RI)- and surface RI-based DNA sensing and one type of bacteria sensing. Furthermore, the effect of glucose oxidase (GOD) immobilization at the sensor head on sensor performance was evaluated for both glucose and synthetic urine solutions with glucose concentration between 0.1% and 10%. Detection sensitivities of the glucose sensors were achieved as low as 0.05%. For chemical sensing, heavy water, ranging from 97% to 10%, and several elemental solutions were monitored by using the FLRD chemical sensors. Bulk index-based FLRD sensing showed that trace elements can be detected in deionized water. For physical sensing, water and cracking sensors were fabricated and embedded into concrete. A partially-etched single-mode fiber (SMF) was embedded into a concrete bar for water monitoring while a bare SMF without any treatment was directly embedded into another concrete bar for monitoring cracks. Furthermore, detection sensitivities of water and crack sensors were investigated as 10 ml water and 0.5 mm surface crack width, respectively. Additionally fiber loop ringdown-fiber Bragg grating temperature sensors were developed in the laboratory; two sensor units for water, crack, and temperature sensing were deployed into a concrete cube in a US Department of Energy test bed (Miami, FL). Multi-sensor applications in a real concrete structure were accomplished by testing the six FLRD sensors. As a final stage, a sensor network was assembled by multiplexing two or three FLRD sensors in series and parallel. Additionally, two FLRD sensors were combined in series and

Nonenzymatic Glucose Sensor Based on In Situ Reduction of Ni/NiO-Graphene Nanocomposite

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Xiaohui Zhang

2016-10-01

Full Text Available Ni/NiO nanoflower modified reduced graphene oxide (rGO nanocomposite (Ni/NiO-rGO was introduced to screen printed electrode (SPE for the construction of a nonenzymatic electrochemical glucose biosensor. The Ni/NiO-rGO nanocomposite was synthesized by an in situ reduction process. Graphene oxide (GO hybrid Nafion sheets first chemical adsorbed Ni ions and assembled on the SPE. Subsequently, GO and Ni ions were reduced by hydrazine hydrate. The electrochemical properties of such a Ni/NiO-rGO modified SPE were carefully investigated. It showed a high activity for electrocatalytic oxidation of glucose in alkaline medium. The proposed nonenzymatic sensor can be utilized for quantification of glucose with a wide linear range from 29.9 μM to 6.44 mM (R = 0.9937 with a low detection limit of 1.8 μM (S/N = 3 and a high sensitivity of 1997 μA/mM∙cm−2. It also exhibited good reproducibility as well as high selectivity.

A new luminol chemiluminescence sensor for glucose based on pH-dependent graphene oxide.

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Hao, Minjia; Liu, Na; Ma, Zhanfang

2013-08-07

In this study, graphene oxide (GO) was found to catalyze the luminol-O2 reaction, which yielded a novel chemiluminescence (CL). Remarkably, the CL emission could be tuned by modulating the pH of the GO dispersion. Transmission electron microscopy, CL spectra, electron spin resonance spectra studies were carried out to investigate the CL mechanism. The results indicate that the CL emission was attributed to the intrinsic catalytic effect of GO acting as the radical generation proliferators and electron transfer accelerators. Based on the GO catalyzed luminol-O2 system, we successfully developed a new CL sensor to detect glucose. Under the optimized conditions, glucose could be assayed in the range of 0.05 mM to 5 mM with a detection limit of 0.044 mM. For the detection of clinical serum samples, it is well consistent with the data determined by commercially available method in hospital, indicating that the new CL method provides a possible application for the detection of glucose in clinical diagnostics.

Tattoo-based noninvasive glucose monitoring: a proof-of-concept study.

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Bandodkar, Amay J; Jia, Wenzhao; Yardımcı, Ceren; Wang, Xuan; Ramirez, Julian; Wang, Joseph

2015-01-06

We present a proof-of-concept demonstration of an all-printed temporary tattoo-based glucose sensor for noninvasive glycemic monitoring. The sensor represents the first example of an easy-to-wear flexible tattoo-based epidermal diagnostic device combining reverse iontophoretic extraction of interstitial glucose and an enzyme-based amperometric biosensor. In-vitro studies reveal the tattoo sensor's linear response toward physiologically relevant glucose levels with negligible interferences from common coexisting electroactive species. The iontophoretic-biosensing tattoo platform is reduced to practice by applying the device on human subjects and monitoring variations in glycemic levels due to food consumption. Correlation of the sensor response with that of a commercial glucose meter underscores the promise of the tattoo sensor to detect glucose levels in a noninvasive fashion. Control on-body experiments demonstrate the importance of the reverse iontophoresis operation and validate the sensor specificity. This preliminary investigation indicates that the tattoo-based iontophoresis-sensor platform holds considerable promise for efficient diabetes management and can be extended toward noninvasive monitoring of other physiologically relevant analytes present in the interstitial fluid.

"Smart tattoo" glucose biosensors and effect of coencapsulated anti-inflammatory agents.

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Srivastava, Rohit; Jayant, Rahul Dev; Chaudhary, Ayesha; McShane, Michael J

2011-01-01

Minimally invasive glucose biosensors with increased functional longevity form one of the most promising techniques for continuous glucose monitoring. In the present study, we developed a novel nanoengineered microsphere formulation comprising alginate microsphere glucose sensors and anti-inflammatory-drug-loaded alginate microspheres. The formulation was prepared and characterized for size, shape, in vitro drug release, biocompatibility, and in vivo acceptability. Glucose oxidase (GOx)- and Apo-GOx-based glucose sensors were prepared and characterized. Sensing was performed both in distilled water and simulated interstitial body fluid. Layer-by-layer self-assembly techniques were used for preventing drug and sensing chemistry release. Finally, in vivo studies, involving histopathologic examination of subcutaneous tissue surrounding the implanted sensors using Sprague-Dawley rats, were performed to test the suppression of inflammation and fibrosis associated with glucose sensor implantation. The drug formulation showed 100% drug release with in 30 days with zero-order release kinetics. The GOx-based sensors showed good enzyme retention and enzyme activity over a period of 1 month. Apo-GOx-based visible and near-infrared sensors showed good sensitivity and analytical response range of 0-50 mM glucose, with linear range up to 12 mM glucose concentration. In vitro cell line studies proved biocompatibility of the material used. Finally, both anti-inflammatory drugs were successful in controlling the implant-tissue interface by suppressing inflammation at the implant site. The incorporation of anti-inflammatory drug with glucose biosensors shows promise in improving sensor biocompatibility, thereby suggesting potential application of alginate microspheres as "smart tattoo" glucose sensors with increased functional longevity. © 2010 Diabetes Technology Society.

Novel glucose biosensor based on a glassy carbon electrode modified with hollow gold nanoparticles and glucose oxidase

International Nuclear Information System (INIS)

Wang, W.; Ying, S.; Zhang, Z.; Huang, S.

2011-01-01

A novel glucose biosensor is presented as that based on a glassy carbon electrode modified with hollow gold nanoparticles (HGNs) and glucose oxidase. The sensor exhibits a better differential pulse voltammetric response towards glucose than the one based on conventional gold nanoparticles of the same size. This is attributed to the good biological conductivity and biocompatibility of HGNs. Under the optimal conditions, the sensor displays a linear range from 2.0 x 10 -6 to 4.6 x 10 -5 M of glucose, with a detection limit of 1.6 x 10 -6 M (S/N = 3). Good reproducibility, stability and no interference make this biosensor applicable to the determination of glucose in samples such as sports drinks. (author)

Glucose Monitoring System Based on Osmotic Pressure Measurements

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Alexandra LEAL

2011-02-01

Full Text Available This paper presents the design and development of a prototype sensor unit for implementation in a long-term glucose monitoring system suitable for estimating glucose levels in people suffering from diabetes mellitus. The system utilizes osmotic pressure as the sensing mechanism and consists of a sensor prototype that is integrated together with a pre-amplifier and data acquisition unit for both data recording and processing. The sensor prototype is based on an embedded silicon absolute pressure transducer and a semipermeable nanoporous membrane that is enclosed in the sensor housing. The glucose monitoring system facilitates the integration of a low power microcontroller that is combined with a wireless inductive powered communication link. Experimental verification have proven that the system is capable of tracking osmotic pressure changes using albumin as a model compound, and thereby show a proof of concept for novel long term tracking of blood glucose from remote sensor nodes.

O-GlcNAcylation mediates the control of cytosolic phosphoenolpyruvate carboxykinase activity via Pgc1α.

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Pedro Latorre

Full Text Available PGC1α is a coactivator of many transcription factors and cytosolic phosphoenolpyruvate carboxykinase (PCK1 is a key enzyme for gluconeogenesis. PGC1α interacts with the transcription factor PPARγ to stimulate PCK1 expression and thus de novo glucose synthesis. These proteins are not only important for central energy metabolism but also for supplying intermediates for other metabolic pathways, including lipidogenesis and protein synthesis and might therefore be important factors in the ethiopathogenesis of metabolic disorders like diabetes but also in other pathologies like cancer. Since polymorphisms in these proteins have been related to some phenotypic traits in animals like pigs and PGC1α G482S polymorphism increases fat deposition in humans, we have investigated the molecular basis of such effects focusing on a commonly studied polymorphism in pig Pgc1α, which changes a cysteine at position 430 (WT of the protein to a serine (C430S. Biochemical analyses show that Pgc1α WT stimulates higher expression of human PCK1 in HEK293T and HepG2 cells. Paradoxically, Pgc1α WT is less stable than Pgc1α p.C430S in HEK293T cells. However, the study of different post-translational modifications shows a higher O-GlcNAcylation level of Pgc1α p.C430S. This higher O-GlcNAcylation level significantly decreases the interaction between Pgc1α and PPARγ demonstrating the importance of post-translational glycosylation of PGC1α in the regulation of PCK1 activity. This, furthermore, could explain at least in part the observed epistatic effects between PGC1α and PCK1 in pigs.

Nanosensors and nanomaterials for monitoring glucose in diabetes.

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Cash, Kevin J; Clark, Heather A

2010-12-01

Worldwide, diabetes is a rapidly growing problem that is managed at the individual level by monitoring and controlling blood glucose levels to minimize the negative effects of the disease. Because of limitations in diagnostic methods, significant research efforts are focused on developing improved methods to measure glucose. Nanotechnology has impacted these efforts by increasing the surface area of sensors, improving the catalytic properties of electrodes and providing nanoscale sensors. Here, we discuss developments in the past several years on both nanosensors that directly measure glucose and nanomaterials that improve glucose sensor function. Finally, we discuss challenges that must be overcome to apply these developments in the clinic. Copyright © 2010 Elsevier Ltd. All rights reserved.

Construction and characterization of nonenzymatic glucose sensor from recycling of Co, Cu and Mn from spent batteries

International Nuclear Information System (INIS)

Selvatici, Livia Serra; Favalessa, Luiza Botan; Loureiro, Eduardo dos Santos; Dixini, Pedro Vitor Morbach; Freitas, Marcos Benedito Jose de; Celante, Vinicius Guilherme

2016-01-01

Full text: In this work, Co, Cu and MnO 2 films were synthesized from the electrochemical recycling of spent Li-ion and alkaline batteries and applied as non-enzymatic electrochemical sensors for glucose determination in aqueous solution. The batteries were dismantled, physically separated into different constituents and leached solution of acetic acid 3:1 v/v. The films were synthesized in potentiostatic condition with E =-0.90 V and fixed charge density of 10 C.cm -2 on glassy carbon substrate with area equal to 3.0 mm 2 . Measurements of X-ray diffraction showed the Co structures (111), Cu (101) and MnO 2 (110). By analysis of scanning electron microscopy, a homogeneous coating of the surface was observed without the presence of surface irregularities. For energy dispersive X-ray, Co, Cu, Mn and O were observed. These films were subsequently used in determining glucose in aqueous solution by measures of successive voltammetric cycles in solutions of concentration varying from 0.1 to 0.5 g/l. These solutions were related to anodic peak current (I peak ), relative oxidation of glucose in glucolactona, the concentration of the solution, obtaining a linear correlation coefficient of 0.989. The sensor stability was measured in 20 voltammetric cycles in each solution, obtaining a correlation coefficient equal to 0978, being stable in the measurements. (author)

Development of a glucose sensor employing quick and easy modification method with mediator for altering electron acceptor preference.

Science.gov (United States)

Hatada, Mika; Loew, Noya; Inose-Takahashi, Yuka; Okuda-Shimazaki, Junko; Tsugawa, Wakako; Mulchandani, Ashok; Sode, Koji

2018-06-01

Enzyme based electrochemical biosensors are divided into three generations according to their type of electron transfer from the cofactors of the enzymes to the electrodes. Although the 3rd generation sensors using direct electron transfer (DET) type enzymes are ideal, the number of enzyme types which possess DET ability is limited. In this study, we report of a glucose sensor using mediator-modified glucose dehydrogenase (GDH), that was fabricated by a new quick-and-easy method using the pre-functionalized amine reactive phenazine ethosulfate (arPES). Thus mediator-modified GDH obtained the ability to transfer electrons to bulky electron acceptors as well as electrodes. The concentration of glucose was successfully measured using electrodes with immobilized PES-modified GDH, without addition of external electron mediators. Therefore, continuous monitoring systems can be developed based on this "2.5th generation" electron transfer principle utilizing quasi-DET. Furthermore, we successfully modified two other diagnostically relevant enzymes, glucoside 3-dehydrogenase and lactate oxidase, with PES. Therefore, various kinds of diagnostic enzymes can achieve quasi-DET ability simply by modification with arPES, suggesting that continuous monitoring systems based on the 2.5th generation principle can be developed for various target molecules. Copyright © 2018 Elsevier B.V. All rights reserved.

Pharmacological Inhibition of O-GlcNAcase Enhances Autophagy in Brain through an mTOR-Independent Pathway.

Science.gov (United States)

Zhu, Yanping; Shan, Xiaoyang; Safarpour, Farzaneh; Erro Go, Nancy; Li, Nancy; Shan, Alice; Huang, Mina C; Deen, Matthew; Holicek, Viktor; Ashmus, Roger; Madden, Zarina; Gorski, Sharon; Silverman, Michael A; Vocadlo, David J

2018-03-05

The glycosylation of nucleocytoplasmic proteins with O-linked N-acetylglucosamine residues (O-GlcNAc) is conserved among metazoans and is particularly abundant within brain. O-GlcNAc is involved in diverse cellular processes ranging from the regulation of gene expression to stress response. Moreover, O-GlcNAc is implicated in various diseases including cancers, diabetes, cardiac dysfunction, and neurodegenerative diseases. Pharmacological inhibition of O-GlcNAcase (OGA), the sole enzyme that removes O-GlcNAc, reproducibly slows neurodegeneration in various Alzheimer's disease (AD) mouse models manifesting either tau or amyloid pathology. These data have stimulated interest in the possibility of using OGA-selective inhibitors as pharmaceuticals to alter the progression of AD. The mechanisms mediating the neuroprotective effects of OGA inhibitors, however, remain poorly understood. Here we show, using a range of methods in neuroblastoma N2a cells, in primary rat neurons, and in mouse brain, that selective OGA inhibitors stimulate autophagy through an mTOR-independent pathway without obvious toxicity. Additionally, OGA inhibition significantly decreased the levels of toxic protein species associated with AD pathogenesis in the JNPL3 tauopathy mouse model as well as the 3×Tg-AD mouse model. These results strongly suggest that OGA inhibitors act within brain through a mechanism involving enhancement of autophagy, which aids the brain in combatting the accumulation of toxic protein species. Our study supports OGA inhibition being a feasible therapeutic strategy for hindering the progression of AD and other neurodegenerative diseases. Moreover, these data suggest more targeted strategies to stimulate autophagy in an mTOR-independent manner may be found within the O-GlcNAc pathway. These findings should aid the advancement of OGA inhibitors within the clinic.

Scanning the available Dictyostelium discoideum proteome for O-linked GlcNAc glycosylation sitesusing neural networks

DEFF Research Database (Denmark)

Gupta, Ramneek; Jung, Eva; Gooley, Andrew A

1999-01-01

Dictyostelium discoideum has been suggested as a eukaryotic model organism for glycobiology studies. Presently, the characteristics of acceptor sites for the N-acetylglucosaminyl-transferases in Dictyostelium discoideum, which link GlcNAc in an alpha linkage to hydroxyl residues, are largely...... unknown. This motivates the development of a species specific method for prediction of O-linked GlcNAc glycosylation sites in secreted and membrane proteins of D. discoideum. The method presented here employs a jury of artificial neural networks. These networks were trained to recognize the sequence...... context and protein surface accessibility in 39 experimentally determined O-alpha-GlcNAc sites found in D. discoideum glycoproteins expressed in vivo. Cross-validation of the data revealed a correlation in which 97% of the glycosylated and nonglycosylated sites were correctly identified. Based...

Glucose sensing based on Pt-MWCNT and MWCNT

Science.gov (United States)

Aryasomayajula, Lavanya; Xie, Jining; Wang, Shouyan; Varadan, Vijay K.

2007-04-01

It is known that multi walled carbon nanotubes (MWCNTs) is an excellent materials for biosensing applications and with the introduction of Pt nanoparticles (Pt-MWCNTs) of about 3nm in diameter in MWCNTs greatly increases the current sensitivity and also the signal to noise ratio. We fabricated the CNT- based glucose sensor by immobilization the bio enzyme, glucose oxidase (GoX), on the Pt-MWCNT and electrode were prepared. The sensor has been tested effectively for both the abnormal blood glucose levels- greater than 6.9 mM and less than 3.5 mM which are the prediabetic and diabetic glucose levels, respectively. The current signal obtained from the Pt-MWCNT was much higher compared to the MWCNT based sensors.

Gibberellic Acid-Stimulated Arabidopsis6 Serves as an Integrator of Gibberellin, Abscisic Acid, and Glucose Signaling during Seed Germination in Arabidopsis.

Science.gov (United States)

Zhong, Chunmei; Xu, Hao; Ye, Siting; Wang, Shiyi; Li, Lingfei; Zhang, Shengchun; Wang, Xiaojing

2015-11-01

The DELLA protein REPRESSOR OF ga1-3-LIKE2 (RGL2) plays an important role in seed germination under different conditions through a number of transcription factors. However, the functions of the structural genes associated with RGL2-regulated germination are less defined. Here, we report the role of an Arabidopsis (Arabidopsis thaliana) cell wall-localized protein, Gibberellic Acid-Stimulated Arabidopsis6 (AtGASA6), in functionally linking RGL2 and a cell wall loosening expansin protein (Arabidopsis expansin A1 [AtEXPA1]), resulting in the control of embryonic axis elongation and seed germination. AtGASA6-overexpressing seeds showed precocious germination, whereas transfer DNA and RNA interference mutant seeds displayed delayed seed germination under abscisic acid, paclobutrazol, and glucose (Glc) stress conditions. The differences in germination rates resulted from corresponding variation in cell elongation in the hypocotyl-radicle transition region of the embryonic axis. AtGASA6 was down-regulated by RGL2, GLUCOSE INSENSITIVE2, and ABSCISIC ACID-INSENSITIVE5 genes, and loss of AtGASA6 expression in the gasa6 mutant reversed the insensitivity shown by the rgl2 mutant to paclobutrazol and the gin2 mutant to Glc-induced stress, suggesting that it is involved in regulating both the gibberellin and Glc signaling pathways. Furthermore, it was found that the promotion of seed germination and length of embryonic axis by AtGASA6 resulted from a promotion of cell elongation at the embryonic axis mediated by AtEXPA1. Taken together, the data indicate that AtGASA6 links RGL2 and AtEXPA1 functions and plays a role as an integrator of gibberellin, abscisic acid, and Glc signaling, resulting in the regulation of seed germination through a promotion of cell elongation. © 2015 American Society of Plant Biologists. All Rights Reserved.

Photo-acoustic sensor based on an inexpensive piezoelectric film transducer and an amplitude-stabilized single-mode external cavity diode laser for in vitro measurements of glucose concentration

Science.gov (United States)

Bayrakli, Ismail; Erdogan, Yasar Kemal

2018-06-01

The present paper focuses on development of a compact photo-acoustic sensor using inexpensive components for glucose analysis. An amplitude-stabilized wavelength-tunable single-mode external cavity diode laser operating around 1050 nm was realized and characterized for the use of laser beam as an excitation light source. In the established setup, a fine tuning range of 9 GHz was achieved. The glucose solution was obtained by diluting D-glucose in sterile water. The acoustic signal generated by the optical excitation was detected via a chip piezoelectric film transducer. A detection limit of 50 mM (900 mg/dl) was achieved. The device may be of great interest for its applications in medicine and health monitoring. The sensor is promising for non-invasive in vivo glucose measurements from interstitial fluid.

An Enzymatic Glucose Sensor Composed of Carbon-Coated Nano Tin Sulfide

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Ren-Jei Chung

2017-02-01

Full Text Available In this study, a biosensor, based on a glucose oxidase (GOx immobilized, carbon-coated tin sulfide (SnS assembled on a glass carbon electrode (GCE was developed, and its direct electrochemistry was investigated. The carbon coated SnS (C-SnS nanoparticle was prepared through a simple two-step process, using hydrothermal and chemical vapor deposition methods. The large reactive surface area and unique electrical potential of C-SnS could offer a favorable microenvironment for facilitating electron transfer between enzymes and the electrode surface. The structure and sensor ability of the proposed GOx/C-SnS electrode were characterized using scanning electron microscopy (SEM, X-ray diffraction (XRD, Raman spectroscopy, UV–vis spectroscopy, Fourier transform infrared spectroscopy (FTIR, and cyclic voltammetry study (CV.

Non-enzymatic glucose detection based on phenylboronic acid modified optical fibers

Science.gov (United States)

Sun, Xiaolan; Li, Nana; Zhou, Bin; Zhao, Wei; Liu, Liyuan; Huang, Chao; Ma, Longfei; Kost, Alan R.

2018-06-01

A non-enzymatic, sensitive glucose sensor was fabricated based on an evanescent wave absorbing optical fiber probe. The optical fiber sensor was functionalized by fixing a poly (phenylboronic acid) (polyPBA) film onto the conical region of the single mode fiber. The reflected light intensity of the polyPBA-functionalized fiber sensor increased proportionally with glucose concentration in the range of 0-60 mM, and the sensor showed good reproducibility and stability. The developed sensor possessed a high sensitivity of 0.1787%/mM and good linearity. The measurement of glucose concentration in human serum was also demonstrated.

Effects of 5 Thio-D-Glucose on cellular adenosine triphosphate levels and deoxyribonucleic acid rejoining in hypoxic and aerobic Chinese hamster cells

International Nuclear Information System (INIS)

Nagle, W.A.; Moss, A.J. Jr.; Roberts, H.G. Jr.; Baker, M.L.

1980-01-01

Intracellular adenosine triphosphate (ATP) levels were measured in both hypoxic and aerobic cultures of V79 Chinese hamster cells treated with 5-thio-D-glucose (5-SH-D-Glc). This glucose analog, a known inhibitor of D-glucose transport and metabolism, reduced ATP in cell cultures allowed to become hypoxic by cell metabolism, but not in aerobic cultures treated similarly. Cells depleted of ATP were unable to rejoin x-ray induced deoxyribonucleic acid (DNA) strand breaks as measured by the alkaline sucrose gradient sedimentation technique. The inference for radiation therapy is that inhibition of glucose metabolism selectively depletes energy reserves in hypoxic cells, rendering these cells more radiosensitive and leading to a more effective tumor treatment

Accuracy of flash glucose monitoring and continuous glucose monitoring technologies: Implications for clinical practice.

Science.gov (United States)

Ajjan, Ramzi A; Cummings, Michael H; Jennings, Peter; Leelarathna, Lalantha; Rayman, Gerry; Wilmot, Emma G

2018-02-01

Continuous glucose monitoring and flash glucose monitoring technologies measure glucose in the interstitial fluid and are increasingly used in diabetes care. Their accuracy, key to effective glycaemic management, is usually measured using the mean absolute relative difference of the interstitial fluid sensor compared to reference blood glucose readings. However, mean absolute relative difference is not standardised and has limitations. This review aims to provide a consensus opinion on assessing accuracy of interstitial fluid glucose sensing technologies. Mean absolute relative difference is influenced by glucose distribution and rate of change; hence, we express caution on the reliability of comparing mean absolute relative difference data from different study systems and conditions. We also review the pitfalls associated with mean absolute relative difference at different glucose levels and explore additional ways of assessing accuracy of interstitial fluid devices. Importantly, much data indicate that current practice of assessing accuracy of different systems based on individualised mean absolute relative difference results has limitations, which have potential clinical implications. Healthcare professionals must understand the factors that influence mean absolute relative difference as a metric for accuracy and look at additional assessments, such as consensus error grid analysis, when evaluating continuous glucose monitoring and flash glucose monitoring systems in diabetes care. This in turn will ensure that management decisions based on interstitial fluid sensor data are both effective and safe.

Integrated process design for biocatalytic synthesis by a Leloir Glycosyltransferase: UDP-glucose production with sucrose synthase.

Science.gov (United States)

Schmölzer, Katharina; Lemmerer, Martin; Gutmann, Alexander; Nidetzky, Bernd

2017-04-01

Nucleotide sugar-dependent ("Leloir") glycosyltransferases (GTs), represent a new paradigm for the application of biocatalytic glycosylations to the production of fine chemicals. However, it remains to be shown that GT processes meet the high efficiency targets of industrial biotransformations. We demonstrate in this study of uridine-5'-diphosphate glucose (UDP-glc) production by sucrose synthase (from Acidithiobacillus caldus) that a holistic process design, involving coordinated development of biocatalyst production, biotransformation, and downstream processing (DSP) was vital for target achievement at ∼100 g scale synthesis. Constitutive expression in Escherichia coli shifted the recombinant protein production mainly to the stationary phase and enhanced the specific enzyme activity to a level (∼480 U/g cell dry weight ) suitable for whole-cell biotransformation. The UDP-glc production had excellent performance metrics of ∼100 g product /L, 86% yield (based on UDP), and a total turnover number of 103 g UDP-glc /g cell dry weight at a space-time yield of 10 g/L/h. Using efficient chromatography-free DSP, the UDP-glc was isolated in a single batch with ≥90% purity and in 73% isolated yield. Overall, the process would allow production of ∼0.7 kg of isolated product/L E. coli bioreactor culture, thus demonstrating how integrated process design promotes the practical use of a GT conversion. Biotechnol. Bioeng. 2017;114: 924-928. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

Glucose ingestion during endurance training does not alter adaptation

DEFF Research Database (Denmark)

Åkerström, Thorbjörn; Fischer, Christian P; Plomgaard, Peter

2009-01-01

, 2) lower citrate synthase (CS) and beta-hydroxyacyl-CoA dehydrogenase (beta-HAD) activity and glycogen content in skeletal muscle, and 3) attenuated endurance performance enhancement in the trained state. To investigate this we studied nine male subjects who performed 10 wk of one-legged knee...... extensor training. They trained one leg while ingesting a 6% glucose solution (Glc) and ingested a sweetened placebo while training the other leg (Plc). The subjects trained their respective legs 2 h at a time on alternate days 5 days a week. Endurance training increased peak power (P(max)) and time...

Antioxidant and Anti-Inflammatory Effects of Blueberry Anthocyanins on High Glucose-Induced Human Retinal Capillary Endothelial Cells

Directory of Open Access Journals (Sweden)

Wuyang Huang

2018-01-01

Full Text Available Blueberries possess abundant anthocyanins, which benefit eye health. The purpose of this study was to explore the protective functional role of blueberry anthocyanin extract (BAE and its predominant constituents, malvidin (Mv, malvidin-3-glucoside (Mv-3-glc, and malvidin-3-galactoside (Mv-3-gal, on high glucose- (HG- induced injury in human retinal capillary endothelial cells (HRCECs. The results showed that BAE, Mv, Mv-3-glc, and Mv-3-gal enhanced cell viability (P<0.05 versus the HG group at 24 h; decreased the reactive oxygen species (ROS, P<0.01 versus the HG group both at 24 and 48 h; and increased the enzyme activity of catalase (CAT and superoxide dismutase (SOD (P<0.05 versus the HG group both at 24 and 48 h. Mv could greatly inhibit HG-induced Nox4 expression both at 24 and 48 h (P<0.05, while BAE and Mv-3-gal downregulated Nox4 only at 48 h (P<0.05. Mv, Mv-3-glc, and Mv-3-gal also changed nitric oxide (NO levels (P<0.05. BAE and Mv-3-glc also influenced angiogenesis by decreasing the vascular endothelial cell growth factor (VEGF level and inhibiting Akt pathway (P<0.05. Moreover, Mv and Mv-3-glc inhibited HG-induced intercellular adhesion molecule-1 (ICAM-1, P<0.001 and nuclear factor-kappa B (NF-κB (P<0.05. It indicated that blueberry anthocyanins protected HRCECs via antioxidant and anti-inflammatory mechanisms, which could be promising molecules for the development of nutraceuticals to prevent diabetic retinopathy.

New evidence of connections between increased O-GlcNAcylation and inflammasome in the oral mucosa of patients with oral lichen planus.

Science.gov (United States)

Thi Do, T; Phoomak, C; Champattanachai, V; Silsirivanit, A; Chaiyarit, P

2018-04-01

Oral lichen planus (OLP) is considered a chronic inflammatory immune-mediated disease of the oral mucosa. Immunopathogenesis of OLP is thought to be associated with cell-mediated immune dysregulation. O-GlcNAcylation is a form of reversible glycosylation. It has been demonstrated that O-GlcNAcylation promoted nuclear factor kappa B (NF-κB) signalling. Activation of NF-кB can induce expression of nucleotide-binding domain-like receptor family pyrin domain containing 3 (NLRP3) inflammasome, which is a large intracellular multi-protein complex involving an immune response. Dysregulated expression of the NLRP3 inflammasome was reported to be associated with autoinflammatory diseases. No integrative studies between O-GlcNAcylation and NLRP3 inflammasome in OLP patients have been reported. The present study aimed to determine the immunohistochemical expression of O-GlcNAcylation, NF-κB signalling molecules and NLRP3 inflammasome in oral mucosae of OLP patients. Oral tissue samples were collected from 30 OLP patients and 30 healthy individuals. Immunohistochemical staining and analyses of immunostaining scores were performed to evaluate expression of O-GlcNAcylation, NF-κB signalling molecules and NLRP3 inflammasome. According to observations in this study, significantly higher levels of O-GlcNAcylation, NF-κB signalling molecules and NLRP3 inflammasome were demonstrated in OLP patients compared with control subjects (P O-GlcNAcylation, NF-κB signalling molecules and NLRP3 inflammasome were also observed in OLP samples (P O-GlcNAcylation is associated with increased expression of NLRP3 inflammasome via the NF-κB signalling pathway. These findings provide a new perspective on immunopathogenesis of OLP in relation to autoinflammation. © 2017 British Society for Immunology.

Improving the Accuracy of the Water Surface Cover Type in the 30 m FROM-GLC Product

Directory of Open Access Journals (Sweden)

Luyan Ji

2015-10-01

Full Text Available The finer resolution observation and monitoring of the global land cover (FROM-GLC product makes it the first 30 m resolution global land cover product from which one can extract a global water mask. However, two major types of misclassification exist with this product due to spectral similarity and spectral mixing. Mountain and cloud shadows are often incorrectly classified as water since they both have very low reflectance, while more water pixels at the boundaries of water bodies tend to be misclassified as land. In this paper, we aim to improve the accuracy of the 30 m FROM-GLC water mask by addressing those two types of errors. For the first, we adopt an object-based method by computing the topographical feature, spectral feature, and geometrical relation with cloud for every water object in the FROM-GLC water mask, and set specific rules to determine whether a water object is misclassified. For the second, we perform a local spectral unmixing using a two-endmember linear mixing model for each pixel falling in the water-land boundary zone that is 8-neighborhood connected to water-land boundary pixels. Those pixels with big enough water fractions are determined as water. The procedure is automatic. Experimental results show that the total area of inland water has been decreased by 15.83% in the new global water mask compared with the FROM-GLC water mask. Specifically, more than 30% of the FROM-GLC water objects have been relabeled as shadows, and nearly 8% of land pixels in the water-land boundary zone have been relabeled as water, whereas, on the contrary, fewer than 2% of water pixels in the same zone have been relabeled as land. As a result, both the user’s accuracy and Kappa coefficient of the new water mask (UA = 88.39%, Kappa = 0.87 have been substantially increased compared with those of the FROM-GLC product (UA = 81.97%, Kappa = 0.81.

Vertically Aligned Carbon Nanofiber based Biosensor Platform for Glucose Sensor

Energy Technology Data Exchange (ETDEWEB)

Al Mamun, Khandaker A.; Tulip, Fahmida S.; MacArthur, Kimberly; McFarlane, Nicole; Islam, Syed K.; Hensley, Dale

2014-03-01

Vertically aligned carbon nanofibers (VACNFs) have recently become an important tool for biosensor design. Carbon nanofibers (CNF) have excellent conductive and structural properties with many irregularities and defect sites in addition to exposed carboxyl groups throughout their surfaces. These properties allow a better immobilization matrix compared to carbon nanotubes and offer better resolution when compared with the FET-based biosensors. VACNFs can be deterministically grown on silicon substrates allowing optimization of the structures for various biosensor applications. Two VACNF electrode architectures have been employed in this study and a comparison of their performances has been made in terms of sensitivity, sensing limitations, dynamic range, and response time. The usage of VACNF platform as a glucose sensor has been verified in this study by selecting an optimum architecture based on the VACNF forest density. Read More: http://www.worldscientific.com/doi/abs/10.1142/S0129156414500062

Defining carbohydrate specificity of Ricinus communis agglutinin as Gal beta 1-->4GlcNAc (II) > Gal beta 1-->3GlcNAc (I) > Gal alpha 1-->3Gal (B) > Gal beta 1-->3GalNAc (T).

Science.gov (United States)

Wu, J H; Herp, A; Wu, A M

1993-03-01

To define carbohydrate specificity of Ricinus communis agglutinin (RCA1), the combining site of RCA1 was further characterized by quantitative precipitin (QPA) and precipitin-inhibition assays (QPIA). Among the oligosaccharides tested for QPIA, Gal beta 1-->4GlcNAc (II, human blood group type II precursor sequence) was found to be 7.1 times more active than Gal beta 1-->3GalNAc (T, Thomsen-Friedenreich sequence) and about 1.7 times more active than the other three disaccharides tested--Gal beta 1-->4Man, Gal beta 1-->3DAra and Gal beta 1-->6GalNAc. Gal alpha 1-->4Gal, the receptor of the uropathogenic E. coli ligand was 3.6 times less active than the II sequence. These results indicate that the beta 1-->4 linkage of the terminal Gal to subterminal GlcNAc is important as this beta 1-->4GlcNAc sequence is at least 1.6 times more active than other types of disaccharides. Among the glycoproteins examined for QPA, native and desialized bovine submandibular glycoproteins, native and desialized human plasma alpha 1-acid glycoproteins, as well as crude hog stomach mucin and its three mild acid hydrolyzed products reacted well with the lectin. These glycoproteins precipitated over 75% of the lectin nitrogen added indicating that RCA1 has the ability to recognize Gal beta 1-->4/3GlcNAc and/or the related residues at the non-reducing ends and at positions in the interior of the chains. However, Tn (GalNAc alpha 1-->Ser/Thr sequence) rich glycoproteins such as desialized ovine submandibular glycoprotein and desialized armadillo salivary glycoprotein, in which over 90% of the carbohydrate side chains are Tn determinants with none or only a trace of I/II or T determinants, precipitated poorly with RCA1. From the present and previous results obtained, the carbohydrate specificity of RCA1 can be constructed and summarized in decreasing order by lectin determinants as follows: II (Gal beta 1-->4GlcNAc) > I (Gal beta 1-->3GlcNAc) > E (Gal alpha 1-->4Gal) and B (Gal alpha 1-->3Gal

Detection of saliva-range glucose concentrations using organic thin-film transistors

International Nuclear Information System (INIS)

Elkington, D.; Belcher, W. J.; Dastoor, P. C.; Zhou, X. J.

2014-01-01

We describe the development of a glucose sensor through direct incorporation of an enzyme (glucose oxidase) into the gate of an organic thin film transistor (OTFT). We show that glucose diffusion is the key determinant of the device response time and present a mechanism of glucose sensing in these devices that involves protonic doping of the transistor channel via enzymatic oxidation of glucose. The integrated OTFT sensor is sensitive across 4 decades of glucose concentration; a range that encompasses both the blood and salivary glucose concentration levels. As such, this work acts as a proof-of-concept for low-cost printed biosensors for salivary glucose.

Detection of saliva-range glucose concentrations using organic thin-film transistors

Energy Technology Data Exchange (ETDEWEB)

Elkington, D.; Belcher, W. J.; Dastoor, P. C.; Zhou, X. J. [Centre for Organic Electronics, University of Newcastle, Callaghan, New South Wales 2308 (Australia)

2014-07-28

We describe the development of a glucose sensor through direct incorporation of an enzyme (glucose oxidase) into the gate of an organic thin film transistor (OTFT). We show that glucose diffusion is the key determinant of the device response time and present a mechanism of glucose sensing in these devices that involves protonic doping of the transistor channel via enzymatic oxidation of glucose. The integrated OTFT sensor is sensitive across 4 decades of glucose concentration; a range that encompasses both the blood and salivary glucose concentration levels. As such, this work acts as a proof-of-concept for low-cost printed biosensors for salivary glucose.

Optical biosensor optimized for continuous in-line glucose monitoring in animal cell culture.

Science.gov (United States)

Tric, Mircea; Lederle, Mario; Neuner, Lisa; Dolgowjasow, Igor; Wiedemann, Philipp; Wölfl, Stefan; Werner, Tobias

2017-09-01

Biosensors for continuous glucose monitoring in bioreactors could provide a valuable tool for optimizing culture conditions in biotechnological applications. We have developed an optical biosensor for long-term continuous glucose monitoring and demonstrated a tight glucose level control during cell culture in disposable bioreactors. The in-line sensor is based on a commercially available oxygen sensor that is coated with cross-linked glucose oxidase (GOD). The dynamic range of the sensor was tuned by a hydrophilic perforated diffusion membrane with an optimized permeability for glucose and oxygen. The biosensor was thoroughly characterized by experimental data and numerical simulations, which enabled insights into the internal concentration profile of the deactivating by-product hydrogen peroxide. The simulations were carried out with a one-dimensional biosensor model and revealed that, in addition to the internal hydrogen peroxide concentration, the turnover rate of the enzyme GOD plays a crucial role for biosensor stability. In the light of this finding, the glucose sensor was optimized to reach a long functional stability (>52 days) under continuous glucose monitoring conditions with a dynamic range of 0-20 mM and a response time of t 90  ≤ 10 min. In addition, we demonstrated that the sensor was sterilizable with beta and UV irradiation and only subjected to minor cross sensitivity to oxygen, when an oxygen reference sensor was applied. Graphical abstract Measuring setup of a glucose biosensor in a shake flask for continuous glucose monitoring in mammalian cell culture.

The differential expression of BmGlcNAcase2 in strains of Bombyx mori (Lepidoptera: Bombycidae) with different susceptibility to Bombyx mori (Lepidoptera: Bombycidae) nucleopolyhedrovirus infection.

Science.gov (United States)

Hao, Zhu; Quanbing, Ma; Xiaoyong, Liu

2015-01-01

GlcNAcase is a glycosyl hydrolase located in the lysosomes of numerous organisms. Levels of the protein, β-N-acetylglucosaminidase 2 (GlcNAcase2), which is a member of the GlcNAcase family, are different in two strains of the silkworm Bombyx mori that have different resistance to Bombyx mori nucleopolyhedroviruses (BmNPVs). We identified six single-nucleotide differences in the GlcNAcase2 coding sequence between the 306 and NB strains. Five are silent changes, but one is a nonsynonymous mutation. Reverse transcription-polymerase chain reaction analysis showed that GlcNAcase2 mRNA levels in the NB strain were nearly 2.57 times higher compared with those in the 306 strain. In addition, GlcNAcase2 enzyme activity was much higher in the NB strain compared with that in the 306 strain. Together, these results indicate that GlcNAcase2 may be involved in variable BmNPV resistance in B. mori. © The Author 2015. Published by Oxford University Press on behalf of the Entomological Society of America.

Higher O-GlcNAc Levels Are Associated with Defects in Progenitor Proliferation and Premature Neuronal Differentiation during in-Vitro Human Embryonic Cortical Neurogenesis

Directory of Open Access Journals (Sweden)

Shama Parween

2017-12-01

Full Text Available The nutrient responsive O-GlcNAcylation is a dynamic post-translational protein modification found on several nucleocytoplasmic proteins. Previous studies have suggested that hyperglycemia induces the levels of total O-GlcNAcylation inside the cells. Hyperglycemia mediated increase in protein O-GlcNAcylation has been shown to be responsible for various pathologies including insulin resistance and Alzheimer's disease. Since maternal hyperglycemia during pregnancy is associated with adverse neurodevelopmental outcomes in the offspring, it is intriguing to identify the effect of increased protein O-GlcNAcylation on embryonic neurogenesis. Herein using human embryonic stem cells (hESCs as model, we show that increased levels of total O-GlcNAc is associated with decreased neural progenitor proliferation and premature differentiation of cortical neurons, reduced AKT phosphorylation, increased apoptosis and defects in the expression of various regulators of embryonic corticogenesis. As defects in proliferation and differentiation during neurodevelopment are common features of various neurodevelopmental disorders, increased O-GlcNAcylation could be one mechanism responsible for defective neurodevelopmental outcomes in metabolically compromised pregnancies such as diabetes.

Wearable Contact Lens Biosensors for Continuous Glucose Monitoring Using Smartphones.

Science.gov (United States)

Elsherif, Mohamed; Hassan, Mohammed Umair; Yetisen, Ali K; Butt, Haider

2018-05-17

Low-cost, robust, and reusable continuous glucose monitoring systems that can provide quantitative measurements at point-of-care settings is an unmet medical need. Optical glucose sensors require complex and time-consuming fabrication processes, and their readouts are not practical for quantitative analyses. Here, a wearable contact lens optical sensor was created for the continuous quantification of glucose at physiological conditions, simplifying the fabrication process and facilitating smartphone readouts. A photonic microstructure having a periodicity of 1.6 μm was printed on a glucose-selective hydrogel film functionalized with phenylboronic acid. Upon binding with glucose, the microstructure volume swelled, which modulated the periodicity constant. The resulting change in the Bragg diffraction modulated the space between zero- and first-order spots. A correlation was established between the periodicity constant and glucose concentration within 0-50 mM. The sensitivity of the sensor was 12 nm mM -1 , and the saturation response time was less than 30 min. The sensor was integrated with commercial contact lenses and utilized for continuous glucose monitoring using smartphone camera readouts. The reflected power of the first-order diffraction was measured via a smartphone application and correlated to the glucose concentrations. A short response time of 3 s and a saturation time of 4 min was achieved in the continuous monitoring mode. Glucose-sensitive photonic microstructures may have applications in point-of-care continuous monitoring devices and diagnostics at home settings.

Promiscuous activity of ER glucosidase II discovered through donor specificity analysis of UGGT

Energy Technology Data Exchange (ETDEWEB)

Miyagawa, Atsushi, E-mail: miyagawa.atsushi@nitech.ac.jp [RIKEN Advanced Science Institute, Wako, Saitama 351-0198 (Japan); Department of Materials Science and Engineering, Nagoya Institute of Technology, Showa-Ku, Nagoya 466-8555 (Japan); Totani, Kiichiro [Department of Materials and Life Science, Seikei University, Musashino, Tokyo 180-8633 (Japan); Matsuo, Ichiro [Department of Chemistry and Chemical Biology, Gunma University, Kiryu, Gunma 376-8515 (Japan); Ito, Yukishige, E-mail: yukito@riken.jp [RIKEN Advanced Science Institute, Wako, Saitama 351-0198 (Japan); ERATO Japan Science and Technology Agency, 2-1 Hirosawa, Wako, Saitama 351-0198 (Japan)

2010-12-17

Research highlights: {yields} UGGT has a narrow donor specificity. {yields} UGGT gave several non-natural high-mannose-type glycans. {yields} G-II has a promiscuous activity as broad specificity hexosidase. -- Abstract: In glycoprotein quality control system in the endoplasmic reticulum (ER), UGGT (UDP-glucose:glycoprotein glucosyltransferase) and glucosidase II (G-II) play key roles. UGGT serves as a glycoprotein folding sensor by virtue of its unique specificity to glucosylate glycoproteins at incompletely folded stage. By using various UDP-Glc analogues, we first analyzed donor specificity of UGGT, which was proven to be rather narrow. However, marginal activity was observed with UDP-galactose and UDP-glucuronic acid as well as with 3-, 4- and 6-deoxy glucose analogues to give corresponding transfer products. Intriguingly, G-II smoothly converted all of them back to Man{sub 9}GlcNAc{sub 2}, providing an indication that G-II has a promiscuous activity as a broad specificity hexosidase.

Purification and identification of O-GlcNAc-modified peptides using phosphate-based alkyne CLICK chemistry in combination with titanium dioxide chromatography and mass spectrometry

DEFF Research Database (Denmark)

Parker, Benjamin L; Gupta, Pankaj; Cordwell, Stuart

2011-01-01

A selective method for the enrichment of O-GlcNAcylated peptides using a novel CLICK chemistry reagent is described. Peptides modified by O-GlcNAc were enzymatically labeled with N-azidoacetylgalactosamine. The azide was then reacted with a phospho-alkyne using CLICK chemistry and O-GlcNAcGalNAzPO4...

Accuracy of subcutaneous continuous glucose monitoring in critically ill adults: improved sensor performance with enhanced calibrations.

Science.gov (United States)

Leelarathna, Lalantha; English, Shane W; Thabit, Hood; Caldwell, Karen; Allen, Janet M; Kumareswaran, Kavita; Wilinska, Malgorzata E; Nodale, Marianna; Haidar, Ahmad; Evans, Mark L; Burnstein, Rowan; Hovorka, Roman

2014-02-01

Accurate real-time continuous glucose measurements may improve glucose control in the critical care unit. We evaluated the accuracy of the FreeStyle(®) Navigator(®) (Abbott Diabetes Care, Alameda, CA) subcutaneous continuous glucose monitoring (CGM) device in critically ill adults using two methods of calibration. In a randomized trial, paired CGM and reference glucose (hourly arterial blood glucose [ABG]) were collected over a 48-h period from 24 adults with critical illness (mean±SD age, 60±14 years; mean±SD body mass index, 29.6±9.3 kg/m(2); mean±SD Acute Physiology and Chronic Health Evaluation score, 12±4 [range, 6-19]) and hyperglycemia. In 12 subjects, the CGM device was calibrated at variable intervals of 1-6 h using ABG. In the other 12 subjects, the sensor was calibrated according to the manufacturer's instructions (1, 2, 10, and 24 h) using arterial blood and the built-in point-of-care glucometer. In total, 1,060 CGM-ABG pairs were analyzed over the glucose range from 4.3 to 18.8 mmol/L. Using enhanced calibration median (interquartile range) every 169 (122-213) min, the absolute relative deviation was lower (7.0% [3.5, 13.0] vs. 12.8% [6.3, 21.8], P<0.001), and the percentage of points in the Clarke error grid Zone A was higher (87.8% vs. 70.2%). Accuracy of the Navigator CGM device during critical illness was comparable to that observed in non-critical care settings. Further significant improvements in accuracy may be obtained by frequent calibrations with ABG measurements.

Mesoporous Nickel Oxide (NiO) Nanopetals for Ultrasensitive Glucose Sensing

Science.gov (United States)

Mishra, Suryakant; Yogi, Priyanka; Sagdeo, P. R.; Kumar, Rajesh

2018-01-01

Glucose sensing properties of mesoporous well-aligned, dense nickel oxide (NiO) nanostructures (NSs) in nanopetals (NPs) shape grown hydrothermally on the FTO-coated glass substrate has been demonstrated. The structural study based investigations of NiO-NPs has been carried out by X-ray diffraction (XRD), electron and atomic force microscopies, energy dispersive X-ray (EDX), and X-ray photospectroscopy (XPS). Brunauer-Emmett-Teller (BET) measurements, employed for surface analysis, suggest NiO's suitability for surface activity based glucose sensing applications. The glucose sensor, which immobilized glucose on NiO-NPs@FTO electrode, shows detection of wide range of glucose concentrations with good linearity and high sensitivity of 3.9 μA/μM/cm2 at 0.5 V operating potential. Detection limit of as low as 1 μΜ and a fast response time of less than 1 s was observed. The glucose sensor electrode possesses good anti-interference ability, stability, repeatability & reproducibility and shows inert behavior toward ascorbic acid (AA), uric acid (UA) and dopamine acid (DA) making it a perfect non-enzymatic glucose sensor.

Self-assembly of palladium nanoparticles on functional TiO{sub 2} nanotubes for a nonenzymatic glucose sensor

Energy Technology Data Exchange (ETDEWEB)

Chen, Xianlan [School of Science, Honghe University, Mengzi, Yunnan 661100 (China); College of Chemistry, Qishan Campus, Fuzhou University, Fuzhou, Fujian 350108 (China); Fujian Key Lab of Medical Instrument & Pharmaceutical Technology, Yishan Campus, Fuzhou University, Fuzhou, Fujian 350002 (China); Li, Gang; Zhang, Guowei [School of Science, Honghe University, Mengzi, Yunnan 661100 (China); Hou, Keyu [College of Chemistry, Qishan Campus, Fuzhou University, Fuzhou, Fujian 350108 (China); Fujian Key Lab of Medical Instrument & Pharmaceutical Technology, Yishan Campus, Fuzhou University, Fuzhou, Fujian 350002 (China); Pan, Haibo, E-mail: hbpan@fzu.edu.cn [College of Chemistry, Qishan Campus, Fuzhou University, Fuzhou, Fujian 350108 (China); Fujian Key Lab of Medical Instrument & Pharmaceutical Technology, Yishan Campus, Fuzhou University, Fuzhou, Fujian 350002 (China); Du, Min [Fujian Key Lab of Medical Instrument & Pharmaceutical Technology, Yishan Campus, Fuzhou University, Fuzhou, Fujian 350002 (China)

2016-05-01

Polydiallyldimethylammonium chloride, PDDA, was used as a stabilizer and linker for functionalized TiO{sub 2} nanotubes (TiO{sub 2} NTs). Self-assembled process with palladium nanoparticles (NPs) was successfully synthesized and used for the oxidation of glucose on glassy carbon electrodes. Based on the voltammetric and amperometric results, Pd NPs efficiently catalyzed the oxidation of glucose at − 0.05 V in the presence of 0.1 M NaCl and showed excellent resistance toward interference poisoning from such interfering species as ascorbic acid, uric acid and urea. To further increase sensitivity, the Pd NPs-PDDA-TiO{sub 2} NTs/GCE was electrochemically treated with H{sub 2}SO{sub 4} and NaOH, the glucose oxidation current was magnified 2.5 times than that before pretreatments due to greatly enhancing the electron transport property of the sensor based on the increased defect sites and surface oxide species. In view of the physiological level of glucose, the wide linear concentration range of glucose (4 × 10{sup −7}–8 × 10{sup −4} M) with a detection limit of 8 × 10{sup −8} M (S/N = 3) was obviously good enough for clinical application. - Highlights: • PDDA was used as a stabilizer and linker for functionalized TiO{sub 2} nanotubes. • Self-assembled process with palladium nanoparticles was synthesized. • After treated both H{sub 2}SO{sub 4} and NaOH, the glucose response was magnified to 2.5 times. • The wide linear concentration range of glucose was obtained with a limit of 8 × 10{sup −8} M.

The O-GlcNAc Transferase Intellectual Disability Mutation L254F Distorts the TPR Helix.

Science.gov (United States)

Gundogdu, Mehmet; Llabrés, Salomé; Gorelik, Andrii; Ferenbach, Andrew T; Zachariae, Ulrich; van Aalten, Daan M F

2018-05-17

O-linked β-N-acetyl- D -glucosamine (O-GlcNAc) transferase (OGT) regulates protein O-GlcNAcylation, an essential post-translational modification that is abundant in the brain. Recently, OGT mutations have been associated with intellectual disability, although it is not understood how they affect OGT structure and function. Using a multi-disciplinary approach we show that the L254F OGT mutation leads to conformational changes of the tetratricopeptide repeats and reduced activity, revealing the molecular mechanisms contributing to pathogenesis. Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.

O-Linked β-N-Acetylglucosaminylation (O-GlcNAcylation) in Primary and Metastatic Colorectal Cancer Clones and Effect of N-Acetyl-β-d-glucosaminidase Silencing on Cell Phenotype and Transcriptome*

Science.gov (United States)

Yehezkel, Galit; Cohen, Liz; Kliger, Adi; Manor, Esther; Khalaila, Isam

2012-01-01

O-Linked β-N-acetylglucosamine (O-GlcNAc) glycosylation is a regulatory post-translational modification occurring on the serine or threonine residues of nucleocytoplasmic proteins. O-GlcNAcylation is dynamically regulated by O-GlcNAc transferase and O-GlcNAcase (OGA), which are responsible for O-GlcNAc addition and removal, respectively. Although O-GlcNAcylation was found to play a significant role in several pathologies such as type II diabetes and neurodegenerative diseases, the role of O-GlcNAcylation in the etiology and progression of cancer remains vague. Here, we followed O-GlcNAcylation and its catalytic machinery in metastatic clones of human colorectal cancer and the effect of OGA knockdown on cellular phenotype and on the transcriptome. The colorectal cancer SW620 metastatic clone exhibited increased O-GlcNAcylation and decreased OGA expression compared with its primary clone, SW480. O-GlcNAcylation elevation in SW620 cells, through RNA interference of OGA, resulted in phenotypic alterations that included acquisition of a fibroblast-like morphology, which coincides with epithelial metastatic progression and growth retardation. Microarray analysis revealed that OGA silencing altered the expression of about 1300 genes, mostly involved in cell movement and growth, and specifically affected metabolic pathways of lipids and carbohydrates. These findings support the involvement of O-GlcNAcylation in various aspects of tumor cell physiology and suggest that this modification may serve as a link between metabolic changes and cancer. PMID:22730328

Mediatorless amperometric bienzyme glucose biosensor based on horseradish peroxidase and glucose oxidase cross-linked to multiwall carbon nanotubes

International Nuclear Information System (INIS)

Xu, Shuxia; Zhou, Shiyi; Zhang, Xinfeng; Qi, Honglan; Zhang, Chengxiao

2014-01-01

We report on a bienzyme-channeling sensor for sensing glucose without the aid of mediator. It was fabricated by cross-linking horseradish peroxidase (HRP) and glucose oxidase (GOx) on a glassy carbon electrode modified with multiwalled carbon nanotubes (MWNTs). The bienzyme was cross-linked with the MWNTs by glutaraldehyde and bovine serum albumin. The MWNTs were employed to accelerate the electron transfer between immobilized HRP and electrode. Glucose was sensed by amperometric reduction of enzymatically generated H 2 O 2 at an applied voltage of −50 mV (vs. Ag/AgCl). Factors influencing the preparation and performance of the bienzyme electrode were investigated in detail. The biosensor exhibited a fast and linear response to glucose in the concentration range from 0.4 to 15 mM, with a detection limit of 0.4 mM. The sensor exhibited good selectivity and durability, with a long-term relative standard deviation of <5 %. Analysis of glucose-spiked human serum samples yielded recoveries between 96 and 101 %. (author)

Novel synthesis and characterization of Ag@TiO2 core shell nanostructure for non-enzymatic glucose sensor

Science.gov (United States)

T, Dayakar; Venkateswara Rao, K.; Vinodkumar, M.; Bikshalu, K.; Chakradhar, B.; Ramachandra Rao, K.

2018-03-01

Ag@TiO2 core-shell nano composite (ATCSNC) were synthesized by using Ocimum tenuiflorum leaves extract through a simple one-step hydrothermal route for Non-enzymatic glucose sensing material. The prepared NCs were characterized and found high crystallinity, red shift absorbance, interface-bonding parameters, rough surface and network like microstructure through XRD, Uv-vis, FTIR, SEM, and TEM. The prepared ATCSNC have been used for fabrication of glassy carbon electrode (GCE) and the same was applied to test its electro catalytic activity of glucose in 0.1 M NaOH. The promising results were recorded for ATCSNC/GCE with a high sensitivity (1968.72 μAm M-1cm-2), wide linear range (1 μM-8.1 mM), good response time (3 s), and excellent low detection limit (0.19 μM, S/N = 3). Furthermore, the designed sensor exhibits admirable stability and reproducibility, as well as attractive achievability for real sample analysis. As such, the proposed ATCSNC could be highly beneficial in the development of sustainable and eco-friendly glucose sensing devices.

Combined radiolysis/GLC as a tool for the investigation of stabilizing mechanisms

International Nuclear Information System (INIS)

Koch, J.; Eckert, W.R.

1977-01-01

Combined radiolysis/GLC was used to prove the chemical incorporation of lauric acid from cadmium laurate into polyvinyl chloride. The samples were exposed for 10 days to x-rays at a dose rate of 0.345 Mrad/hr

Modulation of O-GlcNAc Levels in the Liver Impacts Acetaminophen-Induced Liver Injury by Affecting Protein Adduct Formation and Glutathione Synthesis.

Science.gov (United States)

McGreal, Steven R; Bhushan, Bharat; Walesky, Chad; McGill, Mitchell R; Lebofsky, Margitta; Kandel, Sylvie E; Winefield, Robert D; Jaeschke, Hartmut; Zachara, Natasha E; Zhang, Zhen; Tan, Ee Phie; Slawson, Chad; Apte, Udayan

2018-04-01

Overdose of acetaminophen (APAP) results in acute liver failure. We have investigated the role of a posttranslational modification of proteins called O-GlcNAcylation, where the O-GlcNAc transferase (OGT) adds and O-GlcNAcase (OGA) removes a single β-D-N-acetylglucosamine (O-GlcNAc) moiety, in the pathogenesis of APAP-induced liver injury. Hepatocyte-specific OGT knockout mice (OGT KO), which have reduced O-GlcNAcylation, and wild-type (WT) controls were treated with 300 mg/kg APAP and the development of injury was studied over a time course from 0 to 24 h. OGT KO mice developed significantly lower liver injury as compared with WT mice. Hepatic CYP2E1 activity and glutathione (GSH) depletion following APAP treatment were not different between WT and OGT KO mice. However, replenishment of GSH and induction of GSH biosynthesis genes were significantly faster in the OGT KO mice. Next, male C57BL/6 J mice were treated Thiamet-G (TMG), a specific inhibitor of OGA to induce O-GlcNAcylation, 1.5 h after APAP administration and the development of liver injury was studied over a time course of 0-24 h. TMG-treated mice exhibited significantly higher APAP-induced liver injury. Treatment with TMG did not affect hepatic CYP2E1 levels, GSH depletion, APAP-protein adducts, and APAP-induced mitochondrial damage. However, GSH replenishment and GSH biosynthesis genes were lower in TMG-treated mice after APAP overdose. Taken together, these data indicate that induction in cellular O-GlcNAcylation exacerbates APAP-induced liver injury via dysregulation of hepatic GSH replenishment response.

Progress Toward NLC/GLC Prototype Accelerator Structures

International Nuclear Information System (INIS)

Wang, J

2004-01-01

The accelerator structure groups for NLC (Next Linear Collider) and GLC (Global Linear Colliders) have successfully collaborated on the research and development of a major series of advanced accelerator structures based on room-temperature technology at X-band frequency. The progress in design, simulation, microwave measurement and high gradient tests are summarized in this paper. The recent effort in design and fabrication of the accelerator structure prototype for the main linac is presented in detail including HOM (High Order Mode) suppression and design of HOM couplers and fundamental mode couplers, optimized accelerator cavities as well as plans for future structures

Investigation of the Influence of the As-Grown ZnO Nanorods and Applied Potentials on an Electrochemical Sensor for In-Vitro Glucose Monitoring

Directory of Open Access Journals (Sweden)

Mohammed Marie

2017-01-01

Full Text Available The influence of the as-grown zinc oxide nanorods (ZnO NRs on the fabricated electrochemical sensor for in vitro glucose monitoring were investigated. A direct growth of ZnO NRs was performed on the Si/SiO2/Au electrode, using hydrothermal and sol-gel techniques at low temperatures. The structure, consisting of a Si/SiO2/Au/GOx/Nafion membrane, was considered as a baseline, and it was tested under several applied potential 0.1–0.8 V. The immobilized working electrode, with GOx and a nafion membrane, was characterized amperometrically using a source meter Keithely 2410, and an electrochemical impedance Gamry potentiostat. The sensor exhibited the following: a high sensitivity of ~0.468 mA/cm2 mM, a low detection limit in the order of 166.6 µM, and a fast and sharp response time of around 2 s. The highest sensitivity and the lowest limit of detection were obtained at 0.4 volt, after the growth of ZnO NRs. The highest net sensitivity was obtained after subtracting the sensitivity of the baseline, and it was in the order of 0.315 mA/cm2·mM. The device was tested with a range of glucose concentrations from 1–10 mM, showing a linear line from 3–8 mM, and the device was saturated after exceeding high concentrations of glucose. Such devices can be used for in vitro glucose monitoring, since glucose changes can be accurately detected.

Regression Methods for Ophthalmic Glucose Sensing Using Metamaterials

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Philipp Rapp

2011-01-01

Full Text Available We present a novel concept for in vivo sensing of glucose using metamaterials in combination with automatic learning systems. In detail, we use the plasmonic analogue of electromagnetically induced transparency (EIT as sensor and evaluate the acquired data with support vector machines. The metamaterial can be integrated into a contact lens. This sensor changes its optical properties such as reflectivity upon the ambient glucose concentration, which allows for in situ measurements in the eye. We demonstrate that estimation errors below 2% at physiological concentrations are possible using simulations of the optical properties of the metamaterial in combination with an appropriate electrical circuitry and signal processing scheme. In the future, functionalization of our sensor with hydrogel will allow for a glucose-specific detection which is insensitive to other tear liquid substances providing both excellent selectivity and sensitivity.

Non-Enzymatic Glucose Sensing Using Carbon Quantum Dots Decorated with Copper Oxide Nanoparticles

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Houcem Maaoui

2016-10-01

Full Text Available Perturbations in glucose homeostasis is critical for human health, as hyperglycemia (defining diabetes leads to premature death caused by macrovascular and microvascular complications. However, the simple and accurate detection of glucose in the blood at low cost remains a challenging task, although it is of great importance for the diagnosis and therapy of diabetic patients. In this work, carbon quantum dots decorated with copper oxide nanostructures (CQDs/Cu2O are prepared by a simple hydrothermal approach, and their potential for electrochemical non-enzymatic glucose sensing is evaluated. The proposed sensor exhibits excellent electrocatalytic activity towards glucose oxidation in alkaline solutions. The glucose sensor is characterized by a wide concentration range from 6 µM to 6 mM, a sensitivity of 2.9 ± 0.2 µA·µM−1·cm−2, and a detection limit of 6 µM at a signal-to-noise ratio S/N = 3. The sensors are successfully applied for glucose determination in human serum samples, demonstrating that the CQDs/Cu2O-based glucose sensor satisfies the requirements of complex sample detection with adapted potential for therapeutic diagnostics.

Synthetic Receptors for the High-Affinity Recognition of O-GlcNAc Derivatives

NARCIS (Netherlands)

Rios, Pablo; Carter, Tom S; Mooibroek, Tiddo J; Crump, Matthew P; Lisbjerg, Micke; Pittelkow, Michael; Supekar, Nitin T; Boons, Geert-Jan|info:eu-repo/dai/nl/088245489; Davis, Anthony P

2016-01-01

The combination of a pyrenyl tetraamine with an isophthaloyl spacer has led to two new water-soluble carbohydrate receptors ("synthetic lectins"). Both systems show outstanding affinities for derivatives of N-acetylglucosamine (GlcNAc) in aqueous solution. One receptor binds the methyl glycoside

Synthetic assembly of novel avidin-biotin-GlcNAc (ABG) complex as an attractive bio-probe and its interaction with wheat germ agglutinin (WGA).

Science.gov (United States)

Kumari, Amrita; Koyama, Tetsuo; Hatano, Ken; Matsuoka, Koji

2016-10-01

A tetravalent GlcNAc pendant glycocluster was constructed with terminal biotin through C6 linker. To acquire the multivalent carbohydrate-protein interactions, we synthesized a glycopolymer of tetrameric structure using N-acetyl-d-glucosamine (GlcNAc) as the target carbohydrate by the use of 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (DMT-MM) as coupling reagent, followed by biotin-avidin complexation leading to the formation of glycocluster of avidin-biotin-GlcNAc conjugate (ABG complex). The dynamic light scattering (DLS) system was implied for size detection and to check the binding affinity of GlcNAc conjugate with a WGA lectin we use fluorometric assay by means of specific excitation of tryptophan at λex 295nm and it was found to be very high Ka∼1.39×10(7) M(-1) in case of ABG complex as compared to GlcNAc only Ka∼1.01×10(4) M(-1) with the phenomenon proven to be due to glycocluster effect. Copyright © 2016 Elsevier Inc. All rights reserved.

An In-Line Photonic Biosensor for Monitoring of Glucose Concentrations

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Ala'aldeen Al-Halhouli

2014-08-01

Full Text Available This paper presents two PDMS photonic biosensor designs that can be used for continuous monitoring of glucose concentrations. The first design, the internally immobilized sensor, consists of a reactor chamber, micro-lenses and self-alignment structures for fiber optics positioning. This sensor design allows optical detection of glucose concentrations under continuous glucose flow conditions of 33 µL/h based on internal co-immobilization of glucose oxidase (GOX and horseradish peroxidase (HRP on the internal PDMS surface of the reactor chamber. For this design, two co-immobilization methods, the simple adsorption and the covalent binding (PEG methods were tested. Experiments showed successful results when using the covalent binding (PEG method, where glucose concentrations up to 5 mM with a coefficient of determination (R2 of 0.99 and a limit of detection of 0.26 mM are detectable. The second design is a modified version of the internally immobilized sensor, where a microbead chamber and a beads filling channel are integrated into the sensor. This modification enabled external co-immobilization of enzymes covalently onto functionalized silica microbeads and allows binding a huge amount of HRP and GOX enzymes on the microbeads surfaces which increases the interaction area between immobilized enzymes and the analyte. This has a positive effect on the amount and rate of chemical reactions taking place inside the chamber. The sensor was tested under continuous glucose flow conditions and was found to be able to detect glucose concentrations up to 10 mM with R2 of 0.98 and a limit of detection of 0.7 mM. Such results are very promising for the application in photonic LOC systems used for online analysis.

The Performance and Usability of a Factory-Calibrated Flash Glucose Monitoring System.

Science.gov (United States)

Bailey, Timothy; Bode, Bruce W; Christiansen, Mark P; Klaff, Leslie J; Alva, Shridhara

2015-11-01

The purpose of the study was to evaluate the performance and usability of the FreeStyle(®) Libre™ Flash glucose monitoring system (Abbott Diabetes Care, Alameda, CA) for interstitial glucose results compared with capillary blood glucose results. Seventy-two study participants with type 1 or type 2 diabetes were enrolled by four U.S. clinical sites. A sensor was inserted on the back of each upper arm for up to 14 days. Three factory-only calibrated sensor lots were used in the study. Sensor glucose measurements were compared with capillary blood glucose (BG) results (approximately eight per day) obtained using the BG meter built into the reader (BG reference) and with the YSI analyzer (Yellow Springs Instrument, Yellow Springs, OH) reference tests at three clinic visits (32 samples per visit). Sensor readings were masked to the participants. The accuracy of the results was demonstrated against capillary BG reference values, with 86.7% of sensor results within Consensus Error Grid Zone A. The percentage of readings within Consensus Error Grid Zone A on Days 2, 7, and 14 was 88.4%, 89.2%, and 85.2%, respectively. The overall mean absolute relative difference was 11.4%. The mean lag time between sensor and YSI reference values was 4.5±4.8 min. Sensor accuracy was not affected by factors such as body mass index, age, type of diabetes, clinical site, insulin administration, or hemoglobin A1c. Interstitial glucose measurements with the FreeStyle Libre system were found to be accurate compared with capillary BG reference values, with accuracy remaining stable over 14 days of wear and unaffected by patient characteristics.

Immobilization of glucose oxidase on graphene and cobalt phthalocyanine composite and its application for the determination of glucose.

Science.gov (United States)

Mani, Veerappan; Devasenathipathy, Rajkumar; Chen, Shen-Ming; Huang, Sheng-Tung; Vasantha, V S

2014-11-01

We described a simple and facile chemical reduction strategy for the preparation of graphene (GR)-cobalt phthalocyanine (CoPc) composite and explored it for the enzymatic determination of glucose. CoPc is an active mediator and electrocatalysts for the immobilization of GOx and determination of glucose. However, it is not stable on the electrode surface and also suffers from lack of conductivity. Here, we have employed GR as the suitable support to stabilize CoPc through simple chemical reduction method and the resulting composite has been used for the glucose biosensor application. Scanning electron microscopy, X-ray diffraction and Energy-dispersive X-ray spectroscopy studies confirmed the successful formation of composite. Direct electron transfer of glucose oxidase (GOx) was observed with well defined redox peaks at the formal potential of -0.44 V. The amount of electroactive GOx (Г) and electron transfer rate constant (ks) were calculated to be 3.77×10(-10) mol cm(-2) and 3.57 s(-1), respectively. The fabricated amperometric biosensor detects glucose in wide linear concentration range from 10 μM to 14.8 mM with high sensitivity of 5.0 9μA mM(-1) cm(-2). The sensor offered very low detection limit (LOD) of 1.6 μM. In addition, practical feasibility of the sensor has been explored in screen printing carbon electrode with accurate determination of glucose present in human blood serum and urine samples. Furthermore, the sensor exhibited appreciable stability, repeatability and reproducibility results. Copyright © 2014 Elsevier Inc. All rights reserved.

Nonenzymetic glucose sensing using carbon functionalized carbon doped ZnO nanorod arrays

Science.gov (United States)

Chakraborty, Pinak; Majumder, Tanmoy; Dhar, Saurab; Mondal, Suvra Prakash

2018-04-01

Fabrication of highly sensitive, long stability and low cost glucose sensors are attractive for biomedical applications and food industries. Most of the commercial glucose sensors are based on enzymatic detection which suffers from problems underlying in enzyme activities. Development of high sensitive, enzyme free sensors is a great challenge for next generation glucose sensing applications. In our study Zinc oxide nanorod sensing electrodes have been grown using low cost hydrothermal route and their nonenzymatic glucose sensing properties have been demonstrated with carbon functionalized, carbon doped ZnO nanorods (C-ZnO NRs) in neutral medium (0.1M PBS, pH 7.4) using cyclic voltammetry and amperometry measurements. The C-ZnO NRs electrodes demonstrated glucose sensitivity˜ 13.66 µAmM-1cm-2 in the concentration range 0.7 - 14 mM.

Direct electron transfer of glucose oxidase and dual hydrogen peroxide and glucose detection based on water-dispersible carbon nanotubes derivative

Energy Technology Data Exchange (ETDEWEB)

Chen, Hsiao-Chien [Department of Biochemistry and Molecular Cell Biology, School of Medicine, College of Medicine, Taipei Medical University, 250, Wuxing St., Taipei 11031, Taiwan (China); Tu, Yi-Ming; Hou, Chung-Che [Department of Chemical and Materials Engineering, Chang Gung University, 259 Wen-Hwa 1st Rd., Tao-Yuan 33302, Taiwan (China); Lin, Yu-Chen [Wah Hong industrial Co. Ltd., 6 Lixing St., Guantian Dist., Tainan City 72046,Taiwan (China); Chen, Ching-Hsiang [Graduate Institute of Applied Science and Technology, National Taiwan University of Science and Technology, 43 Keelung Rd., Sec. 4, Taipei 10607, Taiwan (China); Yang, Kuang-Hsuan, E-mail: khy@mail.vnu.edu.tw [Department of Food and Beverage Management, Vanung University, 1, Van Nung Rd., Shuei-Wei Li, Chung-Li City 32061, Taiwan (China)

2015-03-31

Highlights: • Dual hydrogen peroxide and glucose sensor. • Direct electrochemistry of glucose oxidase used MWCNT-Py/GC electrode. • Change sensing function by adjusting pH value. - Abstract: A water-dispersible multi-walled carbon nanotubes (MWCNTs) derivative, MWCNTs-1-one-dihydroxypyridine (MWCNTs-Py) was synthesis via Friedel–Crafts chemical acylation. Raman spectra demonstrated the conjugated level of MWCNTs-Py was retained after this chemical modification. MWCNTs-Py showed dual hydrogen peroxide (H{sub 2}O{sub 2}) and glucose detections without mutual interference by adjusting pH value. It was sensitive to H{sub 2}O{sub 2} in acidic solution and displayed the high performances of sensitivity, linear range, response time and stability; meanwhile it did not respond to H{sub 2}O{sub 2} in neutral solution. In addition, this positively charged MWCNTs-Py could adsorb glucose oxidase (GOD) by electrostatic attraction. MWCNTs-Py-GOD/GC electrode showed the direct electron transfer (DET) of GOD with a pair of well-defined redox peaks, attesting the bioactivity of GOD was retained due to the non-destroyed immobilization. The high surface coverage of active GOD (3.5 × 10{sup −9} mol cm{sup −2}) resulted in exhibiting a good electrocatalytic activity toward glucose. This glucose sensor showed high sensitivity (68.1 μA mM{sup −1} cm{sup −2}) in a linear range from 3 μM to 7 mM in neutral buffer solution. The proposed sensor could distinguish H{sub 2}O{sub 2} and glucose, thus owning high selectivity and reliability.

Direct electron transfer of glucose oxidase and dual hydrogen peroxide and glucose detection based on water-dispersible carbon nanotubes derivative

International Nuclear Information System (INIS)

Chen, Hsiao-Chien; Tu, Yi-Ming; Hou, Chung-Che; Lin, Yu-Chen; Chen, Ching-Hsiang; Yang, Kuang-Hsuan

2015-01-01

Highlights: • Dual hydrogen peroxide and glucose sensor. • Direct electrochemistry of glucose oxidase used MWCNT-Py/GC electrode. • Change sensing function by adjusting pH value. - Abstract: A water-dispersible multi-walled carbon nanotubes (MWCNTs) derivative, MWCNTs-1-one-dihydroxypyridine (MWCNTs-Py) was synthesis via Friedel–Crafts chemical acylation. Raman spectra demonstrated the conjugated level of MWCNTs-Py was retained after this chemical modification. MWCNTs-Py showed dual hydrogen peroxide (H 2 O 2 ) and glucose detections without mutual interference by adjusting pH value. It was sensitive to H 2 O 2 in acidic solution and displayed the high performances of sensitivity, linear range, response time and stability; meanwhile it did not respond to H 2 O 2 in neutral solution. In addition, this positively charged MWCNTs-Py could adsorb glucose oxidase (GOD) by electrostatic attraction. MWCNTs-Py-GOD/GC electrode showed the direct electron transfer (DET) of GOD with a pair of well-defined redox peaks, attesting the bioactivity of GOD was retained due to the non-destroyed immobilization. The high surface coverage of active GOD (3.5 × 10 −9 mol cm −2 ) resulted in exhibiting a good electrocatalytic activity toward glucose. This glucose sensor showed high sensitivity (68.1 μA mM −1 cm −2 ) in a linear range from 3 μM to 7 mM in neutral buffer solution. The proposed sensor could distinguish H 2 O 2 and glucose, thus owning high selectivity and reliability

Chitosan coated on the layers' glucose oxidase immobilized on cysteamine/Au electrode for use as glucose biosensor.

Science.gov (United States)

Zhang, Yawen; Li, Yunqiu; Wu, Wenjian; Jiang, Yuren; Hu, Biru

2014-10-15

A glucose biosensor was developed via direct immobilization of glucose oxidase (GOD) by self-assembled cysteamine monolayer on Au electrode surface followed by coating chitosan on the surface of electrode. In this work, chitosan film was coated on the surface of GOD as a protection film to ensure the stability and biocompatibility of the constructed glucose biosensor. The different application ranges of sensors were fabricated by immobilizing varied layers of GOD. The modified surface film was characterized by a scanning electron microscope (SEM) and the fabrication process of the biosensor was confirmed through electrochemical impedance spectroscopy (EIS) of ferrocyanide. The performance of cyclic voltammetry (CV) in the absence and presence of 25 mM glucose and ferrocenemethanol showed a diffusion-controlled electrode process and reflected the different maximum currents between the different GOD layers. With the developed glucose biosensor, the detection limits of the two linear responses are 49.96 μM and 316.8 μM with the sensitivities of 8.91 μA mM(-1)cm(-2) and 2.93 μA mM(-1)cm(-2), respectively. In addition, good stability (up to 30 days) of the developed biosensor was observed. The advantages of this new method for sensors construction was convenient and different width ranges of detection can be obtained by modified varied layers of GOD. The sensor with two layers of enzyme displayed two current linear responses of glucose. The present work provided a simplicity and novelty method for producing biosensors, which may help design enzyme reactors and biosensors in the future. Copyright © 2014 Elsevier B.V. All rights reserved.

Glucose Binding Protein as a Novel Optical Glucose Nanobiosensor

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Majed DWEIK

2009-11-01

Full Text Available Development of an in vivo optical sensor requires the utilization of Near Infra Red (NIR fluorophores due to their ability to operate within the biological tissue window. Alexa Fluor 750 (AF750 and Alexa Fluor 680 (AF680 were examined as potential NIR fluorophores for an in vivo fluorescence resonance energy transfer (FRET glucose biosensor. AF680 and AF750 found to be a FRET pair and percent energy transfer was calculated. Next, the tested dye pair was utilized in a competitive binding assay in order to detect glucose. Concanavalin A (Con A and dextran have binding affinity, but in the presence of glucose, glucose displaces dextran due to its higher affinity to Con A than dextran. Finally, the percent signal transfer through porcine skin was examined. The results showed with approximately 4.0 mm porcine skin thickness, 1.98 % of the fluorescence was transmitted and captured by the detector.

Metal nanostructures for non-enzymatic glucose sensing

International Nuclear Information System (INIS)

Tee, Si Yin; Teng, Choon Peng; Ye, Enyi

2017-01-01

This review covers the recent development of metal nanostructures in electrochemical non-enzymatic glucose sensing. It highlights a variety of nanostructured materials including noble metals, other transition metals, bimetallic systems, and their hybrid with carbon-based nanomaterials. Particularly, attention is devoted to numerous approaches that have been implemented for improving the sensors performance by tailoring size, shape, composition, effective surface area, adsorption capability and electron-transfer properties. The correlation of the metal nanostructures to the glucose sensing performance is addressed with respect to the linear concentration range, sensitivity and detection limit. In overall, this review provides important clues from the recent scientific achievements of glucose sensor nanomaterials which will be essentially useful in designing better and more effective electrocatalysts for future electrochemical sensing industry. - Highlights: • Overview of recent development of metal nanostructures in electrochemical non-enzymatic glucose sensing. • Special attention is focussed on noble metals, other transition metals, bimetallic systems, and their hybrid with carbon-based nanomaterials. • Merits and limitations of various metal nanostructures in electrochemical non-enzymatic glucose sensing. • Strategies to improve the glucose sensing performance of metal nanostructures as electrocatalysts.

First Clinical Experience with Retrospective Flash Glucose Monitoring (FGM) Analysis in South Africa: Characterizing Glycemic Control with Ambulatory Glucose Profile.

Science.gov (United States)

Distiller, Larry A; Cranston, Iain; Mazze, Roger

2016-11-01

In 2014, an innovative blinded continuous glucose monitoring system was introduced with automated ambulatory glucose profile (AGP) reporting. The clinical use and interpretation of this new technology has not previously been described. Therefore we wanted to understand its use in characterizing key factors related to glycemic control: glucose exposure, variability, and stability, and risk of hypoglycemia in clinical practice. Clinicians representing affiliated diabetes centers throughout South Africa were trained and subsequently were given flash glucose monitoring readers and 2-week glucose sensors to use at their discretion. After patient use, sensor data were collected and uploaded for AGP reporting. Complete data (sensor AGP with corresponding clinical information) were obtained for 50 patients with type 1 (70%) and type 2 diabetes (30%), irrespective of therapy. Aggregated analysis of AGP data comparing patients with type 1 versus type 2 diabetes, revealed that despite similar HbA1c values between both groups (8.4 ± 2 vs 8.6 ± 1.7%, respectively), those with type 2 diabetes had lower mean glucose levels (9.2 ± 3 vs 10.3 mmol/l [166 ± 54 vs 185 mg/dl]) and lower indices of glucose variability (3.0 ± 1.5 vs 5.0 ± 1.9 mmol/l [54 ± 27 vs 90 ± 34.2 mg/dl]). This highlights key areas for future focus. Using AGP, the characteristics of glucose exposure, variability, stability, and hypoglycemia risk and occurrence were obtained within a short time and with minimal provider and patient input. In a survey at the time of the follow-up visit, clinicians indicated that aggregated AGP data analysis provided important new clinical information and insights. © 2016 Diabetes Technology Society.

The quality control of glycoprotein folding in the endoplasmic reticulum, a trip from trypanosomes to mammals

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A.J. Parodi

1998-05-01

Full Text Available The present review deals with the stages of synthesis and processing of asparagine-linked oligosaccharides occurring in the lumen of the endoplasmic reticulum and their relationship to the acquisition by glycoproteins of their proper tertiary structures. Special emphasis is placed on reactions taking place in trypanosomatid protozoa since their study has allowed the detection of the transient glucosylation of glycoproteins catalyzed by UDP-Glc:glycoprotein glucosyltransferase and glucosidase II. The former enzyme has the unique property of covalently tagging improperly folded conformations by catalyzing the formation of protein-linked Glc1Man7GlcNAc2, Glc1Man8GlcNac2 and Glc1Man9GlcNAc2 from the unglucosylated proteins. Glucosyltransferase is a soluble protein of the endoplasmic reticulum that recognizes protein domains exposed in denatured but not in native conformations (probably hydrophobic amino acids and the innermost N-acetylglucosamine unit that is hidden from macromolecular probes in most native glycoproteins. In vivo, the glucose units are removed by glucosidase II. The influence of oligosaccharides in glycoprotein folding is reviewed as well as the participation of endoplasmic reticulum chaperones (calnexin and calreticulin that recognize monoglucosylated species in the same process. A model for the quality control of glycoprotein folding in the endoplasmic reticulum, i.e., the mechanism by which cells recognize the tertiary structure of glycoproteins and only allow transit to the Golgi apparatus of properly folded species, is discussed. The main elements of this control are calnexin and calreticulin as retaining components, the UDP-Glc:glycoprotein glucosyltransferase as a sensor of tertiary structures and glucosidase II as the releasing agent.

Amperometric Bioelectronic Tongue for glucose determination

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Yazan Al-Issa

2015-03-01

Full Text Available An amperometric Bioelectronic Tongue is reported for glucose determination that contains eight sensor electrodes constructed using different metal electrodes (Pt, Au, oxidoreductase enzymes (glucose oxidase, ascorbate oxidase, uricase, and membrane coatings (Nafion, chitosan. The response to varying concentrations of glucose, ascorbic acid, uric acid, and acetaminophen was tested for two models, concentration determination by current density measurements at individual electrodes and concentration determination by a linear regression model for the entire electrode array. The reduced chi-squared for the full array model was found to be about one order of magnitude lower than that for the individual-electrode model. Discrimination of glucose from chemical interference by the other three species is accomplished through a combination of enzyme catalysis, metal electrocatalysis, and membrane surface charge. The benefit of incorporating enzyme electrodes into the sensor array is illustrated by the lower correlation coefficients between different enzyme electrodes relative to non-enzyme coated electrodes. This approach can be more generally applied to detection of other substrates of oxidoreductase enzymes.

Pulse-voltammetric glucose detection at gold junction electrodes.

Science.gov (United States)

Rassaei, Liza; Marken, Frank

2010-09-01

A novel glucose sensing concept based on the localized change or "modulation" in pH within a symmetric gold-gold junction electrode is proposed. A paired gold-gold junction electrode (average gap size ca. 500 nm) is prepared by simultaneous bipotentiostatic electrodeposition of gold onto two closely spaced platinum disk electrodes. For glucose detection in neutral aqueous solution, the potential of the "pH-modulator" electrode is set to -1.5 V vs saturated calomel reference electrode (SCE) to locally increase the pH, and simultaneously, either cyclic voltammetry or square wave voltammetry experiments are conducted at the sensor electrode. A considerable improvement in the sensor electrode response is observed when a normal pulse voltammetry sequence is applied to the modulator electrode (to generate "hydroxide pulses") and the glucose sensor electrode is operated with fixed bias at +0.5 V vs SCE (to eliminate capacitive charging currents). Preliminary data suggest good linearity for the glucose response in the medically relevant 1-10 mM concentration range (corresponding to 0.18-1.8 g L(-1)). Future electroanalytical applications of multidimensional pulse voltammetry in junction electrodes are discussed.

Identification of GIG1, a GlcNAc-Induced Gene in Candida albicans Needed for Normal Sensitivity to the Chitin Synthase Inhibitor Nikkomycin Z▿§

OpenAIRE

Gunasekera, Angelo; Alvarez, Francisco J.; Douglas, Lois M.; Wang, Hong X.; Rosebrock, Adam P.; Konopka, James B.

2010-01-01

The amino sugar N-acetylglucosamine (GlcNAc) is known to be an important structural component of cells from bacteria to humans, but its roles in cell signaling are less well understood. GlcNAc induces two pathways in the human fungal pathogen Candida albicans. One activates cyclic AMP (cAMP) signaling, which stimulates the formation of hyphal cells and the expression of virulence genes, and the other pathway induces genes needed to catabolize GlcNAc. Microarray analysis of gene expression was...

Design, synthesis and structure of new potential electrochemically active boronic acid-based glucose sensors

DEFF Research Database (Denmark)

Norrild, Jens Chr.; Søtofte, Inger

2002-01-01

In the course of our investigations on new boronic acid based carbohydrate sensors three new boronic acids 3, 7 and 11 containing a ferrocene moiety were synthesised. Their design includes an intramolecular B-N bonding motif in order to facilitate binding at physiological pH. We report the synthe......In the course of our investigations on new boronic acid based carbohydrate sensors three new boronic acids 3, 7 and 11 containing a ferrocene moiety were synthesised. Their design includes an intramolecular B-N bonding motif in order to facilitate binding at physiological pH. We report...... the synthesis of the compounds and our investigations on glucose complexation as studied by C-13 NMR spectroscopy. The crystal structure of 2,4,6-tris[2-(N-ferrocenylmethyl-N-methylaminomethyl) phenyl] boroxin (13) (boroxin of boronic acid 3) (boroxin = cyclotriboroxane) was obtained and compared...... with structures obtained of 2,4,6-tris[2-(N,N-dimethylaminomethyl)phenyl]boroxin (14) and 2,2-dimethyl-1,3-diyl[2-(N,N-dimethylaminomethyl)phenyl]boronate (15). The structure of 13 shows the existence of intramolecular B-N bonds in the solid phase....

The EGF repeat-specific O-GlcNAc-transferase Eogt interacts with notch signaling and pyrimidine metabolism pathways in Drosophila.

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Reto Müller

Full Text Available The O-GlcNAc transferase Eogt modifies EGF repeats in proteins that transit the secretory pathway, including Dumpy and Notch. In this paper, we show that the Notch ligands Delta and Serrate are also substrates of Eogt, that mutation of a putative UDP-GlcNAc binding DXD motif greatly reduces enzyme activity, and that Eogt and the cytoplasmic O-GlcNAc transferase Ogt have distinct substrates in Drosophila larvae. Loss of Eogt is larval lethal and disrupts Dumpy functions, but does not obviously perturb Notch signaling. To identify novel genetic interactions with eogt, we investigated dominant modification of wing blister formation caused by knock-down of eogt. Unexpectedly, heterozygosity for several members of the canonical Notch signaling pathway suppressed wing blister formation. And importantly, extensive genetic interactions with mutants in pyrimidine metabolism were identified. Removal of pyrimidine synthesis alleles suppressed wing blister formation, while removal of uracil catabolism alleles was synthetic lethal with eogt knock-down. Therefore, Eogt may regulate protein functions by O-GlcNAc modification of their EGF repeats, and cellular metabolism by affecting pyrimidine synthesis and catabolism. We propose that eogt knock-down in the wing leads to metabolic and signaling perturbations that increase cytosolic uracil levels, thereby causing wing blister formation.

The continuous glucose monitoring sensor in neonatal intensive care

OpenAIRE

Beardsall, K; Ogilvy-Stuart, A; Ahluwalia, J; Thompson, M; Dunger, D

2005-01-01

Objective: To determine the feasibility of continuous glucose monitoring in the very low birthweight baby requiring intensive care, as these infants are known to be at high risk of abnormalities of glucose control.

Sensitive Fluorescent Sensor for Recognition of HIV-1 dsDNA by Using Glucose Oxidase and Triplex DNA

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Yubin Li

2018-01-01

Full Text Available A sensitive fluorescent sensor for sequence-specific recognition of double-stranded DNA (dsDNA was developed on the surface of silver-coated glass slide (SCGS. Oligonucleotide-1 (Oligo-1 was designed to assemble on the surface of SCGS and act as capture DNA, and oligonucleotide-2 (Oligo-2 was designed as signal DNA. Upon addition of target HIV-1 dsDNA (Oligo-3•Oligo-4, signal DNA could bind on the surface of silver-coated glass because of the formation of C•GoC in parallel triplex DNA structure. Biotin-labeled glucose oxidase (biotin-GOx could bind to signal DNA through the specific interaction of biotin-streptavidin, thereby GOx was attached to the surface of SCGS, which was dependent on the concentration of target HIV-1 dsDNA. GOx could catalyze the oxidation of glucose and yield H2O2, and the HPPA can be oxidized into a fluorescent product in the presence of HRP. Therefore, the concentration of target HIV-1 dsDNA could be estimated with fluorescence intensity. Under the optimum conditions, the fluorescence intensity was proportional to the concentration of target HIV-1 dsDNA over the range of 10 pM to 1000 pM, the detection limit was 3 pM. Moreover, the sensor had good sequence selectivity and practicability and might be applied for the diagnosis of HIV disease in the future.

Noninvasive wearable sensor for indirect glucometry.

Science.gov (United States)

Zilberstein, Gleb; Zilberstein, Roman; Maor, Uriel; Righetti, Pier Giorgio

2018-04-02

A noninvasive mini-sensor for blood glucose concentration assessment has been developed. The monitoring is performed by gently pressing a wrist or fingertip onto the chemochromic mixture coating a thin glass or polymer film positioned on the back panel of a smart watch with PPG/HRM (photoplethysmographic/heart rate monitoring sensor). The various chemochromic components measure the absolute values of the following metabolites present in the sweat: acetone, acetone beta-hydroxybutirate, aceto acetate, water, carbon dioxide, lactate anion, pyruvic acid, Na and K salts. Taken together, all these parameters give information about blood glucose concentration, calculated via multivariate analysis based on neural network algorithms built into the sensor. The Clarke Error Grid shows an excellent correlation between data measured by the standard invasive glucose analyser and the present noninvasive sensor, with all points aligned along a 45-degree diagonal and contained almost exclusively in sector A. Graphs measuring glucose levels five times a day (prior, during and after breakfast and prior, during and after lunch), for different individuals (males and females) show a good correlation between the two curves of conventional, invasive meters vs. the noninvasive sensor, with an error of ±15%. This novel, noninvasive sensor for indirect glucometry is fully miniaturized, easy to use and operate and could represent a valid alternative in clinical settings and for individual, personal users, to current, invasive tools. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

The effects of carbon nanotube addition and oxyfluorination on the glucose-sensing capabilities of glucose oxidase-coated carbon fiber electrodes

Energy Technology Data Exchange (ETDEWEB)

Im, Ji Sun; Yun, Jumi; Kim, Jong Gu [Department of Fine Chemical Engineering and Applied Chemistry, BK21-E2 M, Chungnam National University, Daejeon 305-764 (Korea, Republic of); Bae, Tae-Sung [Department of Fine Chemical Engineering and Applied Chemistry, BK21-E2 M, Chungnam National University, Daejeon 305-764 (Korea, Republic of); Korea Basic Science Institute (KBSI), Jeonju 561-756 (Korea, Republic of); Lee, Young-Seak, E-mail: youngslee@cnu.ac.kr [Department of Fine Chemical Engineering and Applied Chemistry, BK21-E2 M, Chungnam National University, Daejeon 305-764 (Korea, Republic of)

2012-01-15

Glucose-sensing electrodes were constructed from carbon fibers by electrospinning and heat treatment. By controlling the pore size, the specific surface area and pore volume of the electrospun carbon fibers were increased for efficient immobilization of the glucose oxidase. Carbon nanotubes were embedded as an electrically conductive additive to improve the electrical property of the porous carbon fibers. In addition, the surface of the porous carbon fibers was modified with hydrophilic functional groups by direct oxyfluorination to increase the affinity between the hydrophobic carbon surface and the hydrophilic glucose oxidase molecules. The porosity of the carbon fibers was improved significantly with approximately 28- and 35-fold increases in the specific surface area and pore volume, respectively. The number of chemical bonds between carbon and oxygen were increased with higher oxygen content during oxyfluorination based on the X-ray photoelectron spectroscopy results. Glucose sensing was carried out by current voltagram and amperometric methods. A high-performance glucose sensor was obtained with high sensitivity and rapid response time as a result of carbon nanotube addition, physical activation and surface modification. The mechanism of the highly sensitive prepared glucose sensor was modeled by an enzyme kinetics study using the Michaelis-Menten equation.

L-Lactic acid production from glucose and xylose with engineered strains of Saccharomyces cerevisiae: aeration and carbon source influence yields and productivities.

Science.gov (United States)

Novy, Vera; Brunner, Bernd; Nidetzky, Bernd

2018-04-11

Saccharomyces cerevisiae, engineered for L-lactic acid production from glucose and xylose, is a promising production host for lignocellulose-to-lactic acid processes. However, the two principal engineering strategies-pyruvate-to-lactic acid conversion with and without disruption of the competing pyruvate-to-ethanol pathway-have not yet resulted in strains that combine high lactic acid yields (Y LA ) and productivities (Q LA ) on both sugar substrates. Limitations seemingly arise from a dependency on the carbon source and the aeration conditions, but the underlying effects are poorly understood. We have recently presented two xylose-to-lactic acid converting strains, IBB14LA1 and IBB14LA1_5, which have the L-lactic acid dehydrogenase from Plasmodium falciparum (pfLDH) integrated at the pdc1 (pyruvate decarboxylase) locus. IBB14LA1_5 additionally has its pdc5 gene knocked out. In this study, the influence of carbon source and oxygen on Y LA and Q LA in IBB14LA1 and IBB14LA1_5 was investigated. In anaerobic fermentation IBB14LA1 showed a higher Y LA on xylose (0.27 g g Xyl -1 ) than on glucose (0.18 g g Glc -1 ). The ethanol yields (Y EtOH , 0.15 g g Xyl -1 and 0.32 g g Glc -1 ) followed an opposite trend. In IBB14LA1_5, the effect of the carbon source on Y LA was less pronounced (~ 0.80 g g Xyl -1 , and 0.67 g g Glc -1 ). Supply of oxygen accelerated glucose conversions significantly in IBB14LA1 (Q LA from 0.38 to 0.81 g L -1  h -1 ) and IBB14LA1_5 (Q LA from 0.05 to 1.77 g L -1  h -1 ) at constant Y LA (IBB14LA1 ~ 0.18 g g Glc -1 ; IBB14LA1_5 ~ 0.68 g g Glc -1 ). In aerobic xylose conversions, however, lactic acid production ceased completely in IBB14LA1 and decreased drastically in IBB14LA1_5 (Y LA aerobic ≤ 0.25 g g Xyl -1 and anaerobic ~ 0.80 g g Xyl -1 ) at similar Q LA (~ 0.04 g L -1  h -1 ). Switching from aerobic to microaerophilic conditions (pO 2  ~ 2%) prevented lactic acid metabolization, observed for

Oxygen consumption and blood flow coupling in human motor cortex during intense finger tapping

DEFF Research Database (Denmark)

Seyedi Vafaee, Manouchehr; Vang, Kim; Bergersen, Linda H

2012-01-01

Rates of cerebral blood flow (CBF) and glucose consumption (CMR(glc)) rise in cerebral cortex during continuous stimulation, while the oxygen-glucose index (OGI) declines as an index of mismatched coupling of oxygen consumption (cerebral metabolic rate of oxygen-CMRO(2)) to CBF and CMR(glc). To t...

Nanomaterials in glucose sensing

CERN Document Server

Burugapalli, Krishna

2013-01-01

The smartness of nano-materials is attributed to their nanoscale and subsequently unique physicochemical properties and their use in glucose sensing has been aimed at improving performance, reducing cost and miniaturizing the sensor and its associated instrumentation. So far, portable (handheld) glucose analysers were introduced for hospital wards, emergency rooms and physicians' offices; single-use strip systems achieved nanolitre sampling for painless and accurate home glucose monitoring; advanced continuous monitoring devices having 2 to 7 days operating life are in clinical and home use; and continued research efforts are being made to develop and introduce increasingly advanced glucose monitoring systems for health as well as food, biotechnology, cell and tissue culture industries. Nanomaterials have touched every aspect of biosensor design and this chapter reviews their role in the development of advanced technologies for glucose sensing, and especially for diabetes. Research shows that overall, nanomat...

Photochemical immobilization of protein on the inner wall of a microchannel and Its application in a glucose sensor

International Nuclear Information System (INIS)

Nakajima, Hizuru; Ishino, Satomi; Masuda, Hironori; Nakagama, Tatsuro; Shimosaka, Takuya; Uchiyama, Katsumi

2006-01-01

A new protein immobilization technique has been developed for patterning enzymes in a specific position inside a microchannel. First, bovine serum albumin (BSA) was adsorbed onto the internal surface of a polydimethylsiloxane microchannel. The microchannel was then filled with the conjugate solution of a photoreactive cross-linker, 4-azido-2,3,5,6-tetrafluorobenzoic acid succinimidyl ester (ATFB-SE), and an enzyme, horseradish peroxidase (HRP). An irradiation by a He-Cd laser activated the azido group of the conjugates and these conjugates became covalently attached to the adsorbed BSA on the microchannel. The enzyme turnover was observed from only the HRP zone. This technique was successfully applied to the enzymatic glucose sensor. Glucose oxidase (GOD) and HRP were sequentially patterned in a single microchannel, i.e., the HRP zone was located downstream from the GOD zone. The calibration curve of a glucose standard solution was linear over the range of 0-128 μM with a correlation coefficient of 0.993. Compared to the traditional method using a 96-well microtiter plate, the present technique on the microchip shortened the reaction time from 30 min to 4.8 s, i.e., to 1/375

Thumb-size ultrasonic-assisted spectroscopic imager for in-situ glucose monitoring as optional sensor of conventional dialyzers

Science.gov (United States)

Nogo, Kosuke; Mori, Keita; Qi, Wei; Hosono, Satsuki; Kawashima, Natsumi; Nishiyama, Akira; Wada, Kenji; Ishimaru, Ichiro

2016-03-01

We proposed the ultrasonic-assisted spectroscopic imaging for the realization of blood-glucose-level monitoring during dialytic therapy. Optical scattering and absorption caused by blood cells deteriorate the detection accuracy of glucose dissolved in plasma. Ultrasonic standing waves can agglomerate blood cells at nodes. In contrast, around anti-node regions, the amount of transmitted light increases because relatively clear plasma appears due to decline the number of blood cells. Proposed method can disperse the transmitted light of plasma without time-consuming pretreatment such as centrifugation. To realize the thumb-size glucose sensor which can be easily attached to dialysis tubes, an ultrasonic standing wave generator and a spectroscopic imager are required to be small. Ultrasonic oscillators are ∅30[mm]. A drive circuit of oscillators, which now size is 41×55×45[mm], is expected to become small. The trial apparatus of proposed one-shot Fourier spectroscopic imager, whose size is 30×30×48[mm], also can be little-finger size in principal. In the experiment, we separated the suspension mixed water and micro spheres (Θ10[mm) into particles and liquid regions with the ultrasonic standing wave (frequency: 2[MHz]). Furthermore, the spectrum of transmitted light through the suspension could be obtained in visible light regions with a white LED.

Glucose de-repression by yeast AMP-activated protein kinase SNF1 is controlled via at least two independent steps.

Science.gov (United States)

García-Salcedo, Raúl; Lubitz, Timo; Beltran, Gemma; Elbing, Karin; Tian, Ye; Frey, Simone; Wolkenhauer, Olaf; Krantz, Marcus; Klipp, Edda; Hohmann, Stefan

2014-04-01

The AMP-activated protein kinase, AMPK, controls energy homeostasis in eukaryotic cells but little is known about the mechanisms governing the dynamics of its activation/deactivation. The yeast AMPK, SNF1, is activated in response to glucose depletion and mediates glucose de-repression by inactivating the transcriptional repressor Mig1. Here we show that overexpression of the Snf1-activating kinase Sak1 results, in the presence of glucose, in constitutive Snf1 activation without alleviating glucose repression. Co-overexpression of the regulatory subunit Reg1 of the Glc-Reg1 phosphatase complex partly restores glucose regulation of Snf1. We generated a set of 24 kinetic mathematical models based on dynamic data of Snf1 pathway activation and deactivation. The models that reproduced our experimental observations best featured (a) glucose regulation of both Snf1 phosphorylation and dephosphorylation, (b) determination of the Mig1 phosphorylation status in the absence of glucose by Snf1 activity only and (c) a regulatory step directing active Snf1 to Mig1 under glucose limitation. Hence it appears that glucose de-repression via Snf1-Mig1 is regulated by glucose via at least two independent steps: the control of activation of the Snf1 kinase and directing active Snf1 to inactivating its target Mig1. © 2014 FEBS.

Non-invasive optical detection of glucose in cell culture nutrient medium

Science.gov (United States)

Cote, Gerald L.

1993-01-01

The objective of the proposed research was to begin the development of a non-invasive optical sensor for measuring glucose concentration in the output medium of cell cultures grown in a unique NASA bioreactor referred to as an integrated rotating-wall vessel (IRWV). The input, a bovine serum based nutrient media, has a known glucose concentration. The cells within the bioreactor digest a portion of the glucose. Thus, the non-invasive optical sensor is needed to monitor the decrease in glucose due to cellular consumption since the critical parameters for sustained cellular productivity are glucose and pH. Previous glucose sensing techniques have used chemical reactions to quantify the glucose concentration. Chemical reactions, however, cannot provide for continuous, real time, non-invasive measurement as is required in this application. Our effort while in the fellowship program was focused on the design, optical setup, and testing of one bench top prototype non-invasive optical sensor using a mid-infrared absorption spectroscopy technique. Glucose has a fundamental vibrational absorption peak in the mid-infrared wavelength range at 9.6 micron. Preliminary absorption data using a CO2 laser were collected at this wavelength for water based glucose solutions at different concentrations and one bovine serum based nutrient medium (GTSF) with added glucose. The results showed near linear absorption responses for the glucose-in-water data with resolutions as high at 108 mg/dl and as low as 10 mg/dl. The nutrient medium had a resolution of 291 mg/dl. The variability of the results was due mainly to thermal and polarization drifts of the laser while the decrease in sensitivity to glucose in the nutrient medium was expected due to the increase in the number of confounders present in the nutrient medium. A multispectral approach needs to be used to compensate for these confounders. The CO2 laser used for these studies was wavelength tunable (9.2 to 10.8 micrometers), however

Fiber Optic Chemical Sensors

Science.gov (United States)

1993-10-01

31, 1980. Koyama, Masao and Sato , Yuichi, "Improved Enzyme Sensor for Glucose with an Ultra-Filtration Membrane and Immobilized Glucose Oxidase...ion Process," American Laboratory, AVO, AIS, 54-59, 1989. Kaihara, Mikio ; Mametsuka, Hiroaki; Gunji, Naoki; Iwata, Hideo and Gohshi,, "New Dilution

Adipogenesis stimulates the nuclear localization of EWS with an increase in its O-GlcNAc glycosylation in 3T3-L1 cells

International Nuclear Information System (INIS)

Li, Qiang; Kamemura, Kazuo

2014-01-01

Highlights: • The majority of EWS localizes stably in the cytosol in 3T3-L1 preadipocytes. • Adipogenic stimuli induce the nuclear localization of EWS. • Adipogenesis promotes O-GlcNAcylation of EWS. • O-GlcNAcylation stimulates the recruitment of EWS to the nuclear periphery. - Abstract: Although the Ewing sarcoma (EWS) proto-oncoprotein is found in the nucleus and cytosol and is associated with the cell membrane, the regulatory mechanisms of its subcellular localization are still unclear. Here we found that adipogenic stimuli induce the nuclear localization of EWS in 3T3-L1 cells. Tyrosine phosphorylation in the C-terminal PY-nuclear localization signal of EWS was negative throughout adipogenesis. Instead, an adipogenesis-dependent increase in O-linked β-N-acetylglucosamine (O-GlcNAc) glycosylation of EWS was observed. Pharmacological inactivation of O-GlcNAcase in preadipocytes promoted perinuclear localization of EWS. Our findings suggest that the nuclear localization of EWS is partly regulated by the glycosylation

Woven electrochemical fabric-based test sensors (WEFTS): a new class of multiplexed electrochemical sensors.

Science.gov (United States)

Choudhary, Tripurari; Rajamanickam, G P; Dendukuri, Dhananjaya

2015-05-07

We present textile weaving as a new technique for the manufacture of miniature electrochemical sensors with significant advantages over current fabrication techniques. Biocompatible silk yarn is used as the material for fabrication instead of plastics and ceramics used in commercial sensors. Silk yarns are coated with conducting inks and reagents before being handloom-woven as electrodes into patches of fabric to create arrays of sensors, which are then laminated, cut and packaged into individual sensors. Unlike the conventionally used screen-printing, which results in wastage of reagents, yarn coating uses only as much reagent and ink as required. Hydrophilic and hydrophobic yarns are used for patterning so that sample flow is restricted to a small area of the sensor. This simple fluidic control is achieved with readily available materials. We have fabricated and validated individual sensors for glucose and hemoglobin and a multiplexed sensor, which can detect both analytes. Chronoamperometry and differential pulse voltammetry (DPV) were used to detect glucose and hemoglobin, respectively. Industrial quantities of these sensors can be fabricated at distributed locations in the developing world using existing skills and manufacturing facilities. We believe such sensors could find applications in the emerging area of wearable sensors for chemical testing.

Electrochemical Glucose Sensors—Developments Using Electrostatic Assembly and Carbon Nanotubes for Biosensor Construction

Directory of Open Access Journals (Sweden)

Mark R. Anderson

2010-09-01

Full Text Available In 1962, Clark and Lyons proposed incorporating the enzyme glucose oxidase in the construction of an electrochemical sensor for glucose in blood plasma. In their application, Clark and Lyons describe an electrode in which a membrane permeable to glucose traps a small volume of solution containing the enzyme adjacent to a pH electrode, and the presence of glucose is detected by the change in the electrode potential that occurs when glucose reacts with the enzyme in this volume of solution. Although described nearly 50 years ago, this seminal development provides the general structure for constructing electrochemical glucose sensors that is still used today. Despite the maturity of the field, new developments that explore solutions to the fundamental limitations of electrochemical glucose sensors continue to emerge. Here we discuss two developments of the last 15 years; confining the enzyme and a redox mediator to a very thin molecular films at electrode surfaces by electrostatic assembly, and the use of electrodes modified by carbon nanotubes (CNTs to leverage the electrocatalytic effect of the CNTs to reduce the oxidation overpotential of the electrode reaction or for the direct electron transport to the enzyme.

Vascular O-GlcNAcylation augments reactivity to constrictor stimuli by prolonging phosphorylated levels of the myosin light chain

Energy Technology Data Exchange (ETDEWEB)

Lima, V.V. [Instituto de Ciências Biológicas e da Saúde, Universidade Federal de Mato Grosso, Barra do Garças, MT (Brazil); Lobato, N.S.; Filgueira, F.P. [Curso de Medicina, Setor de Fisiologia Humana, Universidade Federal de Goiás, Jataí, GO (Brazil); Webb, R.C. [Department of Physiology, Georgia Regents University, Augusta, GA (United States); Tostes, R.C. [Departamento de Farmacologia, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP (Brazil); Giachini, F.R. [Instituto de Ciências Biológicas e da Saúde, Universidade Federal de Mato Grosso, Barra do Garças, MT (Brazil)

2014-08-15

O-GlcNAcylation is a modification that alters the function of numerous proteins. We hypothesized that augmented O-GlcNAcylation levels enhance myosin light chain kinase (MLCK) and reduce myosin light chain phosphatase (MLCP) activity, leading to increased vascular contractile responsiveness. The vascular responses were measured by isometric force displacement. Thoracic aorta and vascular smooth muscle cells (VSMCs) from rats were incubated with vehicle or with PugNAc, which increases O-GlcNAcylation. In addition, we determined whether proteins that play an important role in the regulation of MLCK and MLCP activity are directly affected by O-GlcNAcylation. PugNAc enhanced phenylephrine (PE) responses in rat aortas (maximal effect, 14.2±2 vs 7.9±1 mN for vehicle, n=7). Treatment with an MLCP inhibitor (calyculin A) augmented vascular responses to PE (13.4±2 mN) and abolished the differences in PE-response between the groups. The effect of PugNAc was not observed when vessels were preincubated with ML-9, an MLCK inhibitor (7.3±2 vs 7.5±2 mN for vehicle, n=5). Furthermore, our data showed that differences in the PE-induced contractile response between the groups were abolished by the activator of AMP-activated protein kinase (AICAR; 6.1±2 vs 7.4±2 mN for vehicle, n=5). PugNAc increased phosphorylation of myosin phosphatase target subunit 1 (MYPT-1) and protein kinase C-potentiated inhibitor protein of 17 kDa (CPI-17), which are involved in RhoA/Rho-kinase-mediated inhibition of myosin phosphatase activity. PugNAc incubation produced a time-dependent increase in vascular phosphorylation of myosin light chain and decreased phosphorylation levels of AMP-activated protein kinase, which decreased the affinity of MLCK for Ca{sup 2+}/calmodulin. Our data suggest that proteins that play an important role in the regulation of MLCK and MLCP activity are directly affected by O-GlcNAcylation, favoring vascular contraction.

Design of nanostructured-based glucose biosensors

Science.gov (United States)

Komirisetty, Archana; Williams, Frances; Pradhan, Aswini; Konda, Rajini B.; Dondapati, Hareesh; Samantaray, Diptirani

2012-04-01

This paper presents the design of glucose sensors that will be integrated with advanced nano-materials, bio-coatings and electronics to create novel devices that are highly sensitive, inexpensive, accurate, and reliable. In the work presented, a glucose biosensor and its fabrication process flow have been designed. The device is based on electrochemical sensing using a working electrode with bio-functionalized zinc oxide (ZnO) nano-rods. Among all metal oxide nanostructures, ZnO nano-materials play a significant role as a sensing element in biosensors due to their properties such as high isoelectric point (IEP), fast electron transfer, non-toxicity, biocompatibility, and chemical stability which are very crucial parameters to achieve high sensitivity. Amperometric enzyme electrodes based on glucose oxidase (GOx) are used due to their stability and high selectivity to glucose. The device also consists of silicon dioxide and titanium layers as well as platinum working and counter electrodes and a silver/silver chloride reference electrode. Currently, the biosensors are being fabricated using the process flow developed. Once completed, the sensors will be bio-functionalized and tested to characterize their performance, including their sensitivity and stability.

Layer-by-Layer Assembly of Glucose Oxidase on Carbon Nanotube Modified Electrodes.

Science.gov (United States)

Suroviec, Alice H

2017-01-01

The use of enzymatically modified electrodes for the detection of glucose or other non-electrochemically active analytes is becoming increasingly common. Direct heterogeneous electron transfer to glucose oxidase has been shown to be kinetically difficult, which is why electron transfer mediators or indirect detection is usually used for monitoring glucose with electrochemical sensors. It has been found, however, that electrodes modified with single or multi-walled carbon nanotubes (CNTs) demonstrate fast heterogeneous electron transfer kinetics as compared to that found for traditional electrodes. Incorporating CNTs into the assembly of electrochemical glucose sensors, therefore, affords the possibility of facile electron transfer to glucose oxidase, and a more direct determination of glucose. This chapter describes the methods used to use CNTs in a layer-by-layer structure along with glucose oxidase to produce an enzymatically modified electrode with high turnover rates, increased stability and shelf-life.

Initial investigation of glucose metabolism in mouse brain using enriched 17 O-glucose and dynamic 17 O-MRS.

Science.gov (United States)

Borowiak, Robert; Reichardt, Wilfried; Kurzhunov, Dmitry; Schuch, Christian; Leupold, Jochen; Krafft, Axel Joachim; Reisert, Marco; Lange, Thomas; Fischer, Elmar; Bock, Michael

2017-08-01

In this initial work, the in vivo degradation of 17 O-labeled glucose was studied during cellular glycolysis. To monitor cellular glucose metabolism, direct 17 O-magnetic resonance spectroscopy (MRS) was used in the mouse brain at 9.4 T. Non-localized spectra were acquired with a custom-built transmit/receive (Tx/Rx) two-turn surface coil and a free induction decay (FID) sequence with a short TR of 5.4 ms. The dynamics of labeled oxygen in the anomeric 1-OH and 6-CH 2 OH groups was detected using a Hankel-Lanczos singular value decomposition (HLSVD) algorithm for water suppression. Time-resolved 17 O-MRS (temporal resolution, 42/10.5 s) was performed in 10 anesthetized (1.25% isoflurane) mice after injection of a 2.2 M solution containing 2.5 mg/g body weight of differently labeled 17 O-glucose dissolved in 0.9% physiological saline. From a pharmacokinetic model fit of the H 2 17 O concentration-time course, a mean apparent cerebral metabolic rate of 17 O-labeled glucose in mouse brain of CMR Glc  = 0.07 ± 0.02 μmol/g/min was extracted, which is of the same order of magnitude as a literature value of 0.26 ± 0.06 μmol/g/min reported by 18 F-fluorodeoxyglucose ( 18 F-FDG) positron emission tomography (PET). In addition, we studied the chemical exchange kinetics of aqueous solutions of 17 O-labeled glucose at the C1 and C6 positions with dynamic 17 O-MRS. In conclusion, the results of the exchange and in vivo experiments demonstrate that the C6- 17 OH label in the 6-CH 2 OH group is transformed only glycolytically by the enzyme enolase into the metabolic end-product H 2 17 O, whereas C1- 17 OH ends up in water via direct hydrolysis as well as glycolysis. Therefore, dynamic 17 O-MRS of highly labeled 17 O-glucose could provide a valuable non-radioactive alternative to FDG PET in order to investigate glucose metabolism. Copyright © 2017 John Wiley & Sons, Ltd.

Electrochemical Glucose Biosensor Based on Glucose Oxidase Displayed on Yeast Surface.

Science.gov (United States)

Wang, Hongwei; Lang, Qiaolin; Liang, Bo; Liu, Aihua

2015-01-01

The conventional enzyme-based biosensor requires chemical or physical immobilization of purified enzymes on electrode surface, which often results in loss of enzyme activity and/or fractions immobilized over time. It is also costly. A major advantage of yeast surface display is that it enables the direct utilization of whole cell catalysts with eukaryote-produced proteins being displayed on the cell surface, providing an economic alternative to traditional production of purified enzymes. Herein, we describe the details of the display of glucose oxidase (GOx) on yeast cell surface and its application in the development of electrochemical glucose sensor. In order to achieve a direct electrochemistry of GOx, the entire cell catalyst (yeast-GOx) was immobilized together with multiwalled carbon nanotubes on the electrode, which allowed sensitive and selective glucose detection.

Sensor-integrated polymer actuators for closed-loop drug delivery system

Science.gov (United States)

Xu, Han; Wang, Chunlei; Kulinsky, Lawrence; Zoval, Jim; Madou, Marc

2006-03-01

This work presents manufacturing and testing of a closed-loop drug delivery system where drug release is achieved by an electrochemical actuation of an array of polymeric valves on a set of drug reservoirs. The valves are based on bi-layer structures made of polypyrrole/gold in the shape of a flap that is hinged on one side of a valve seat. Drugs stored in the underlying chambers are released by bending the bi-layer flaps back with a small applied bias. These polymeric valves simultaneously function as both drug release components and biological/chemical sensors responding to a specific biological or environmental stimulus. The sensors may send signals to the control module to realize closed-loop control of the drug release. In this study a glucose sensor has been integrated with the polymeric actuators through immobilization of glucose oxidase(GOx) within polypyrrole(PPy) valves. Sensitivities per unit area of the integrated glucose sensor have been measured and compared before and after the actuation of the sensor/actuator PPy/DBS/GOx film. Other sensing parameters such as linear range and response time were discussed as well. Using an array of these sensor/actuator cells, the amount of released drug, e.g. insulin, can be precisely controlled according to the surrounding glucose concentration detected by the glucose sensor. Activation of these reservoirs can be triggered either by the signal from the sensor, or by the signal from the operator. This approach also serves as the initial step to use the proposed system as an implantable drug delivery platform in the future.

Glucose biosensor based on glucose oxidase immobilized at gold nanoparticles decorated graphene-carbon nanotubes.

Science.gov (United States)

Devasenathipathy, Rajkumar; Mani, Veerappan; Chen, Shen-Ming; Huang, Sheng-Tung; Huang, Tsung-Tao; Lin, Chun-Mao; Hwa, Kuo-Yuan; Chen, Ting-Yo; Chen, Bo-Jun

2015-10-01

Biopolymer pectin stabilized gold nanoparticles were prepared at graphene and multiwalled carbon nanotubes (GR-MWNTs/AuNPs) and employed for the determination of glucose. The formation of GR-MWNTs/AuNPs was confirmed by scanning electron microscopy, X-ray diffraction, UV-vis and FTIR spectroscopy methods. Glucose oxidase (GOx) was successfully immobilized on GR-MWNTs/AuNPs film and direct electron transfer of GOx was investigated. GOx exhibits highly enhanced redox peaks with formal potential of -0.40 V (vs. Ag/AgCl). The amount of electroactive GOx and electron transfer rate constant were found to be 10.5 × 10(-10) mol cm(-2) and 3.36 s(-1), respectively, which were significantly larger than the previous reports. The fabricated amperometric glucose biosensor sensitively detects glucose and showed two linear ranges: (1) 10 μM - 2 mM with LOD of 4.1 μM, (2) 2 mM - 5.2 mM with LOD of 0.95 mM. The comparison of the biosensor performance with reported sensors reveals the significant improvement in overall sensor performance. Moreover, the biosensor exhibited appreciable stability, repeatability, reproducibility and practicality. The other advantages of the fabricated biosensor are simple and green fabrication approach, roughed and stable electrode surface, fast in sensing and highly reproducible. Copyright © 2015 Elsevier Inc. All rights reserved.

Enhanced non-enzymatic glucose sensing based on copper nanoparticles decorated nitrogen-doped graphene.

Science.gov (United States)

Jiang, Ding; Liu, Qian; Wang, Kun; Qian, Jing; Dong, Xiaoya; Yang, Zhenting; Du, Xiaojiao; Qiu, Baijing

2014-04-15

Copper nanoparticles (NPs) decorated nitrogen-doped graphene (Cu-N-G) was prepared by a facile thermal treatment, and further employed as a novel sensing material for fabricating the sensitive non-enzymatic glucose sensor. Compared with pure Cu NPs, the Cu-N-G showed enhanced electrocatalytic activity to glucose oxidation due to the integration of N-G, which exhibited the oxidation peak current of glucose ca. 23-fold higher than that of pure Cu NPs. The presented sensor showed excellent performances for glucose detection including wide linear range of 0.004-4.5 mM, low detection limit (1.3 μM, S/N=3), high sensitivity (48.13 μA mM(-1)), fast response time (doped graphene as enhanced materials in fabricating sensors for chemical and biochemical analysis. © 2013 Published by Elsevier B.V.

Facile synthesis of a silver nanoparticles/polypyrrole nanocomposite for non-enzymatic glucose determination.

Science.gov (United States)

Poletti Papi, Maurício A; Caetano, Fabio R; Bergamini, Márcio F; Marcolino-Junior, Luiz H

2017-06-01

The present work describes the synthesis of a new conductive nanocomposite based on polypyrrole (PPy) and silver nanoparticles (PPy-AgNP) based on a facile reverse microemulsion method and its application as a non-enzymatic electrochemical sensor for glucose detection. Focusing on the best sensor performance, all experimental parameters used in the synthesis of nanocomposite were optimized based on its electrochemical response for glucose. Characterization of the optimized material by FT-IR, cyclic voltammetry, and DRX measurements and TEM images showed good monodispersion of semispherical Ag nanoparticles capped by PPy structure, with size average of 12±5nm. Under the best analytical conditions, the proposed sensor exhibited glucose response in linear dynamic range of 25 to 2500μmolL -1 , with limit of detection of 3.6μmolL -1 . Recovery studies with human saliva samples varying from 99 to 105% revealed the accuracy and feasibility of a non-enzymatic electrochemical sensor for glucose determination by easy construction and low-cost. Copyright © 2017 Elsevier B.V. All rights reserved.

Tetrahymena pyriformis cells are deficient in all mannose-P-dolichol-dependent mannosyltransferases but not in mannose-P-dolichol synthesis

International Nuclear Information System (INIS)

Yagodnik, C.; de la Canal, L.; Parodi, A.J.

1987-01-01

Cells of the ciliated protozoan Tetrahymena pyriformis incubated with [ 14 C]glucose were found to synthesize Man-P-dolichol and Glc-P-dolichol, as well as Glc 3 Man 5 GlcNAc 2 -P-P-dolichol, the latter being the main and largest lipid derivative formed. The missing mannose residues were those known to be transferred from Man-P-dolichol in other systems. Formation of Man-P-dolichol and of dolichol-P-P-oligosaccharides containing up to five mannose units was detected in cell-free assays containing protozoan membranes, rat liver dolichol-P, unlabeled Man/sub 4-9/GlcNAc 2 -P-P-dolichol from pig liver, and GDP-[ 14 C]Man. Under exactly the same conditions but with UDP-[ 14 C]Glc instead of GDP-[ 14 C]Man, Glc-P-dolichol and dolichol-P-P-oligosaccharides containing five mannose and one to three glucose residues were formed in the absence of the pig liver compounds. In the presence of the latter, dolichol-P-P derivatives containing nine mannose and one to three glucose units were also synthesized. It is concluded that T. pyriformis cells are deficient in all Man-P-dolichol-dependent mannosyltransferases but not in Man-P-dolichol synthesis. The role of the latter compound in this microorganism is unknown

Plain to point network reduced graphene oxide - activated carbon composites decorated with platinum nanoparticles for urine glucose detection

Science.gov (United States)

Hossain, Mohammad Faruk; Park, Jae Y.

2016-02-01

In this study, a hydrothermal technique was applied to synthesize glucose-treated reduced graphene oxide-activated carbon (GRGO/AC) composites. Platinum nanoparticles (PtNP) were electrochemically deposited on the modified GRGO/AC surface, and chitosan-glucose oxidase (Chit-GOx) composites and nafion were integrated onto the modified surface of the working electrode to prepare a highly sensitive glucose sensor. The fabricated biosensor exhibited a good amperometric response to glucose in the detection range from 0.002 mM to 10 mM, with a sensitivity of 61.06 μA/mMcm2, a short response time (4 s) and a low detection limit of 2 μM (signal to noise ratio is 3). The glucose sensor exhibited a negligible response to interference and good stability. In addition, the glucose levels in human urine were tested in order to conduct a practical assessment of the proposed sensor, and the results indicate that the sensor had superior urine glucose recognition. These results thus demonstrate that the noble nano-structured electrode with a high surface area and electrocatalytic activity offers great promise for use in urine glucose sensing applications.

A Disposable Tear Glucose Biosensor—Part 4

Science.gov (United States)

Engelschall, Erica; Lan, Kenneth; Shah, Pankti; Saez, Neil; Maxwell, Stephanie; Adamson, Teagan; Abou-Eid, Michelle; McAferty, Kenyon; Patel, Dharmendra R.; Cook, Curtiss B.

2014-01-01

Objective: A prototype tear glucose (TG) sensor was tested in New Zealand white rabbits to assess eye irritation, blood glucose (BG) and TG lag time, and correlation with BG. Methods: A total of 4 animals were used. Eye irritation was monitored by Lissamine green dye and analyzed using image analysis software. Lag time was correlated with an oral glucose load while recording TG and BG readings. Correlation between TG and BG were plotted against one another to form a correlation diagram, using a Yellow Springs Instrument (YSI) and self-monitoring of blood glucose as the reference measurements. Finally, TG levels were calculated using analytically derived expressions. Results: From repeated testing carried over the course of 12 months, little to no eye irritation was detected. TG fluctuations over time visually appeared to trace the same pattern as BG with an average lag times of 13 minutes. TG levels calculated from the device current measurements ranged from 4 to 20 mg/dL and correlated linearly with BG levels of 75-160 mg/dL (TG = 0.1723 BG = 7.9448 mg/dL; R2 = .7544). Conclusion: The first steps were taken toward preliminary development of a sensor for self-monitoring of tear glucose (SMTG). No conjunctival irritation in any of the animals was noted. Lag time between TG and BG was found to be noticeable, but a quantitative modeling to correlate lag time in this study is unnecessary. Measured currents from the sensors and the calculated TG showed promising correlation to BG levels. Previous analytical bench marking showed BG and TG levels consistent with other literature. PMID:24876546

A novel CuO-N-doped graphene nanocomposite-based hybrid electrode for the electrochemical detection of glucose

Science.gov (United States)

Felix, Sathiyanathan; Kollu, Pratap; Jeong, Soon Kwan; Grace, Andrews Nirmala

2017-10-01

We report a catalyst of N-doped graphene CuO nanocomposite, for the non-enzymatic electrocatalytic oxidation of glucose. The hybrid nanocomposite was synthesized by copper sulfate, cetyl ammonium bromide and graphite as starting materials. The synthesized composites were characterized with the techniques like X-ray diffraction, field emission scanning electron microscopy, transmission electron microscope to study the crystalline phase and morphological structure. Based on this composite, a non-enzymatic glucose sensor was constructed. Cyclic voltammetry and chronoamperometry methods were done to investigate the electrocatalytic properties of glucose in alkaline medium. For glucose detection, the fabricated sensor showed a linear response over a wide range of concentration from 3 to 1000 µM, with sensitivity of 2365.7 µA mM-1 cm-2 and a fast response time of 5 s. The designed sensor exhibited negligible current response to the normal concentration of common interferents in the presence of glucose. All these favorable advantages of the fabricated glucose sensor suggest that it may have good potential application in biological samples, food and other related areas.

Analytical modeling of glucose biosensors based on carbon nanotubes.

Science.gov (United States)

Pourasl, Ali H; Ahmadi, Mohammad Taghi; Rahmani, Meisam; Chin, Huei Chaeng; Lim, Cheng Siong; Ismail, Razali; Tan, Michael Loong Peng

2014-01-15

In recent years, carbon nanotubes have received widespread attention as promising carbon-based nanoelectronic devices. Due to their exceptional physical, chemical, and electrical properties, namely a high surface-to-volume ratio, their enhanced electron transfer properties, and their high thermal conductivity, carbon nanotubes can be used effectively as electrochemical sensors. The integration of carbon nanotubes with a functional group provides a good and solid support for the immobilization of enzymes. The determination of glucose levels using biosensors, particularly in the medical diagnostics and food industries, is gaining mass appeal. Glucose biosensors detect the glucose molecule by catalyzing glucose to gluconic acid and hydrogen peroxide in the presence of oxygen. This action provides high accuracy and a quick detection rate. In this paper, a single-wall carbon nanotube field-effect transistor biosensor for glucose detection is analytically modeled. In the proposed model, the glucose concentration is presented as a function of gate voltage. Subsequently, the proposed model is compared with existing experimental data. A good consensus between the model and the experimental data is reported. The simulated data demonstrate that the analytical model can be employed with an electrochemical glucose sensor to predict the behavior of the sensing mechanism in biosensors.

Towards a continuous glucose monitoring system using tunable quantum cascade lasers

Science.gov (United States)

Haase, Katharina; Müller, Niklas; Petrich, Wolfgang

2018-02-01

We present a reagent-free approach for long-term continuous glucose monitoring (cgm) of liquid samples using midinfrared absorption spectroscopy. This method could constitute an alternative to enzymatic glucose sensors in order to manage the widespread disease of Diabetes. In order to acquire spectra of the liquid specimen, we use a spectrally tunable external-cavity (EC-) quantum cascade laser (QCL) as radiation source in combination with a fiber-based in vitro sensor setup. Hereby we achieve a glucose sensitivity in pure glucose solutions of 3 mg/dL (RMSEP). Furthermore, the spectral tunability of the EC-QCL enables us to discriminate glucose from other molecules. We exemplify this by detecting glucose among other saccharides with an accuracy of 8 mg/dL (within other monosaccharides, RMSEVC) and 14 mg/dL (within other mono- and disaccharides, RMSECV). Moreover, we demonstrate a characterization of the significance of each wavenumber for an accurate prediction of glucose among other saccharides using an evolutionary algorithm. We show, that by picking 10 distinct wavenumbers we can achieve comparable accuracies to the use of a complete spectrum.

On-chip highly sensitive saliva glucose sensing using multilayer films composed of single-walled carbon nanotubes, gold nanoparticles, and glucose oxidase

Directory of Open Access Journals (Sweden)

Wenjun Zhang

2015-06-01

Full Text Available It is very important for human health to rapidly and accurately detect glucose levels in biological environments, especially for diabetes mellitus. We proposed a simple, highly sensitive, accurate, convenient, low-cost, and disposable glucose biosensor on a single chip. A working (sensor electrode, a counter electrode, and a reference electrode are integrated on a single chip through micro-fabrication. The working electrode is functionalized through a layer-by-layer (LBL assembly of single-walled carbon nanotubes (SWNTs and multilayer films composed of chitosan (CS, gold nanoparticles (GNp, and glucose oxidase (GOx to obtain high sensitivity and accuracy. The glucose sensor has following features: (1 direct electron transfer between GOx and the electrode surface; (2 on-a-chip; (3 glucose detection down to 0.1 mg/dL (5.6 μM; (4 good sensing linearity over 0.017–0.81 mM; (5 high sensitivity (61.4 μA/mM-cm2 with a small reactive area (8 mm2; (6 fast response; (7 high reproducibility and repeatability; (8 reliable and accurate saliva glucose detection. Thus, this disposable biosensor will be an alternative for real time tracking of glucose levels from body fluids, e.g. saliva, in a noninvasive, pain-free, accurate, and continuous way. In addition to being used as a disposable glucose biosensor, it also provides a suitable platform for on-chip electrochemical sensing for other chemical agents and biomolecules.

Synthesis of tremella-like CoS and its application in sensing of hydrogen peroxide and glucose

Energy Technology Data Exchange (ETDEWEB)

Wu, Wenqin; Yu, Beibei [Hubei Collaborative Innovation Center for Advanced Organic Chemical Materials, Key Laboratory for the Synthesis and Application of Organic Functional Molecules, Ministry of Education, College of Chemistry & Chemical Engineering, Hubei University, Wuhan 430062 (China); Wu, Huimin, E-mail: whm267@hubu.edu.cn [Hubei Collaborative Innovation Center for Advanced Organic Chemical Materials, Key Laboratory for the Synthesis and Application of Organic Functional Molecules, Ministry of Education, College of Chemistry & Chemical Engineering, Hubei University, Wuhan 430062 (China); Wang, Shengfu; Xia, Qinghua [Hubei Collaborative Innovation Center for Advanced Organic Chemical Materials, Key Laboratory for the Synthesis and Application of Organic Functional Molecules, Ministry of Education, College of Chemistry & Chemical Engineering, Hubei University, Wuhan 430062 (China); Ding, Yu [College of Chemistry and Materials Science, Hubei Engineering University, Xiaogan 432000 (China)

2017-01-01

Different phases of cobalt sulfides have been fabricated by one-pot hydrothermal method. Comparing all of the prepared materials, and the results revealed that CoS was the most conductive and could accelerate electron transfer. The CoS presented tremella-like and excellent catalytic activities towards hydrogen peroxide and glucose. The sensor based on CoS performed amperometric sensing of hydrogen peroxide in a linear range between 5.00 μM and 14.82 mM. Meanwhile, sensing of glucose with double-linear range, one is between 5.00 μM and 1.10 mM, the other is between 1.20 mM and 10.20 mM. These due to the fact that more and more intermediate species absorb onto electrode surface with increasing the concentration of glucose, which limit the following glucose oxidation. Furthermore, the hydrogen peroxide and glucose sensors based on tremella-like CoS also exhibited excellent selectivity, stability, and reproducibility. Thus, the sensor showed potential utilities in hydrogen peroxide and glucose detection. - Highlights: • Tremella-like CoS was prepared by an environmentally friendly hydrothermal method. • The CoS exhibited excellent catalytic activity towards hydrogen peroxide and glucose. • The sensors based on CoS can be applied to detect real samples.

The use of two continuous glucose sensors during and after surgery

NARCIS (Netherlands)

Vriesendorp, T. M.; DeVries, J. H.; Holleman, F.; Dzoljic, M.; Hoekstra, J. B. L.

2005-01-01

BACKGROUND: Maintaining plasma glucose between 80 and 120 mg/dL is beneficial for patients admitted to a surgical intensive care unit, but requires frequent glucose monitoring to ensure adequacy of treatment and detection of hypoglycemia. We examined whether continuous glucose sensing is feasible

Inhibition of GlcNAc-Processing Glycosidases by C-6-Azido-NAG-Thiazoline and Its Derivatives

Czech Academy of Sciences Publication Activity Database

Krejzová, Jana; Šimon, Petr; Ježová-Kalachová, Lubica; Kulik, Natallia; Bojarová, Pavla; Marhol, Petr; Pelantová, Helena; Cvačka, Josef; Ettrich, Rüdiger; Slámová, Kristýna; Křen, Vladimír

2014-01-01

Roč. 19, č. 3 (2014), s. 3471-3488 ISSN 1420-3049 Institutional support: RVO:61388971 ; RVO:67179843 ; RVO:61388963 Keywords : NAG-thiazoline * enzyme inhibition * O-GlcNAcase Subject RIV: CE - Biochemistry Impact factor: 2.416, year: 2014

Preparation and characterization of nickel oxide nanoparticles and their application in glucose and methanol sensing

Directory of Open Access Journals (Sweden)

Mahsa Hasanzadeh

2015-03-01

Full Text Available In this work, a low cost glucose and methanol nonenzymatic sensor was prepared using nickel oxide (NiO nanofilm electrodeposited on a bare Cu electrode. Electrochemical deposition was assisted with cetyl trimethylammonium bromide (CTAB as a template. Scanning electron microscopy (SEM was applied to observe the surface morphology of the modified electrode. Cyclic voltammetry (CV and amperometry techniques were used to study the electrocatalytic behavior of NiO porous film in glucose and methanol detection. For glucose sensing, the electrode showed a linear relationship in the concentration range of 0.01-2.14 mM with a low limit of detection (LOD 1.7 µM (signal/noise ratio (S/N=3. Moreover, high sensitivities of 4.02 mA mM−1 cm−2 and 0.38 mA mM−1 cm−2 respectively in glucose and methanol monitoring suggested the modified electrode as an excellent sensor. The NiO-Cu modified electrode was relatively insensitive to common biological interferers. This sensor possessed good poison resistance towards chloride ions, and long term stability and significant selectivity towards glucose and methanol. Finally the proposed sensor was successfully applied for determination of glucose in human blood serum samples.

Nano-yarn carbon nanotube fiber based enzymatic glucose biosensor

International Nuclear Information System (INIS)

Zhu Zhigang; Burugapalli, Krishna; Moussy, Francis; Song, Wenhui; Li Yali; Zhong Xiaohua

2010-01-01

A novel brush-like electrode based on carbon nanotube (CNT) nano-yarn fiber has been designed for electrochemical biosensor applications and its efficacy as an enzymatic glucose biosensor demonstrated. The CNT nano-yarn fiber was spun directly from a chemical-vapor-deposition (CVD) gas flow reaction using a mixture of ethanol and acetone as the carbon source and an iron nano-catalyst. The fiber, 28 μm in diameter, was made of bundles of double walled CNTs (DWNTs) concentrically compacted into multiple layers forming a nano-porous network structure. Cyclic voltammetry study revealed a superior electrocatalytic activity for CNT fiber compared to the traditional Pt-Ir coil electrode. The electrode end tip of the CNT fiber was freeze-fractured to obtain a unique brush-like nano-structure resembling a scale-down electrical 'flex', where glucose oxidase (GOx) enzyme was immobilized using glutaraldehyde crosslinking in the presence of bovine serum albumin (BSA). An outer epoxy-polyurethane (EPU) layer was used as semi-permeable membrane. The sensor function was tested against a standard reference electrode. The sensitivities, linear detection range and linearity for detecting glucose for the miniature CNT fiber electrode were better than that reported for a Pt-Ir coil electrode. Thermal annealing of the CNT fiber at 250 deg. C for 30 min prior to fabrication of the sensor resulted in a 7.5 fold increase in glucose sensitivity. The as-spun CNT fiber based glucose biosensor was shown to be stable for up to 70 days. In addition, gold coating of the electrode connecting end of the CNT fiber resulted in extending the glucose detection limit to 25 μM. To conclude, superior efficiency of CNT fiber for glucose biosensing was demonstrated compared to a traditional Pt-Ir sensor.

Nano-yarn carbon nanotube fiber based enzymatic glucose biosensor

Science.gov (United States)

Zhu, Zhigang; Song, Wenhui; Burugapalli, Krishna; Moussy, Francis; Li, Ya-Li; Zhong, Xiao-Hua

2010-04-01

A novel brush-like electrode based on carbon nanotube (CNT) nano-yarn fiber has been designed for electrochemical biosensor applications and its efficacy as an enzymatic glucose biosensor demonstrated. The CNT nano-yarn fiber was spun directly from a chemical-vapor-deposition (CVD) gas flow reaction using a mixture of ethanol and acetone as the carbon source and an iron nano-catalyst. The fiber, 28 µm in diameter, was made of bundles of double walled CNTs (DWNTs) concentrically compacted into multiple layers forming a nano-porous network structure. Cyclic voltammetry study revealed a superior electrocatalytic activity for CNT fiber compared to the traditional Pt-Ir coil electrode. The electrode end tip of the CNT fiber was freeze-fractured to obtain a unique brush-like nano-structure resembling a scale-down electrical 'flex', where glucose oxidase (GOx) enzyme was immobilized using glutaraldehyde crosslinking in the presence of bovine serum albumin (BSA). An outer epoxy-polyurethane (EPU) layer was used as semi-permeable membrane. The sensor function was tested against a standard reference electrode. The sensitivities, linear detection range and linearity for detecting glucose for the miniature CNT fiber electrode were better than that reported for a Pt-Ir coil electrode. Thermal annealing of the CNT fiber at 250 °C for 30 min prior to fabrication of the sensor resulted in a 7.5 fold increase in glucose sensitivity. The as-spun CNT fiber based glucose biosensor was shown to be stable for up to 70 days. In addition, gold coating of the electrode connecting end of the CNT fiber resulted in extending the glucose detection limit to 25 µM. To conclude, superior efficiency of CNT fiber for glucose biosensing was demonstrated compared to a traditional Pt-Ir sensor.

Fault and meal detection by redundant continuous glucose monitors and the unscented Kalman filter

DEFF Research Database (Denmark)

Mahmoudi, Zeinab; Nørgaard, Kirsten; Poulsen, Niels Kjølstad

2017-01-01

The purpose of this study is to develop a method for detecting and compensating the anomalies of continuous glucose monitoring (CGM) sensors as well as detecting unannounced meals. Both features, sensor fault detection/correction and meal detection, are necessary to have a reliable artificial pan...... is corrupted by PISA. The fault isolator can detect 199 out of 200 unannounced meals. The average change in the glucose concentrations between the meals and the detection time points is 46.3 mg/dL.......The purpose of this study is to develop a method for detecting and compensating the anomalies of continuous glucose monitoring (CGM) sensors as well as detecting unannounced meals. Both features, sensor fault detection/correction and meal detection, are necessary to have a reliable artificial...... from the two fault detectors differentiates between a sensor fault and an unannounced meal appearing as an anomaly in the CGM data. If the fault isolator indicates a sensor fault, a method based on the covariance matching technique tunes the covariance of the measurement noise associated...

Direct determination of glucose, lactate and triglycerides in blood serum by a tunable quantum cascade laser-based mid-IR sensor

Science.gov (United States)

Brandstetter, M.; Volgger, L.; Genner, A.; Jungbauer, C.; Lendl, B.

2013-02-01

This work reports on a compact sensor for fast and reagent-free point-of-care determination of glucose, lactate and triglycerides in blood serum based on a tunable (1030-1230 cm-1) external-cavity quantum cascade laser (EC-QCL). For simple and robust operation a single beam set-up was designed and only thermoelectric cooling was used for the employed laser and detector. Full computer control of analysis including liquid handling and data analysis facilitated routine measurements. A high optical pathlength (>100 μm) is a prerequisite for robust measurements in clinical practice. Hence, the optimum optical pathlength for transmission measurements in aqueous solution was considered in theory and experiment. The experimentally determined maximum signal-to-noise ratio (SNR) was around 140 μm for the QCL blood sensor and around 50 μm for a standard FT-IR spectrometer employing a liquid nitrogen cooled mercury cadmium telluride (MCT) detector. A single absorption spectrum was used to calculate the analyte concentrations simultaneously by using a partial-least-squares (PLS) regression analysis. Glucose was determined in blood serum with a prediction error (RMSEP) of 6.9 mg/dl and triglycerides with an error of cross-validation (RMSECV) of 17.5 mg/dl in a set of 42 different patients. In spiked serum samples the lactate concentration could be determined with an RMSECV of 8.9 mg/dl.