Search results for: Glucose oxidase andMicrocontroller.
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 160

Search results for: Glucose oxidase andMicrocontroller.

160 Performance Evaluation of an Amperometric Biosensor using a Simple Microcontroller based Data Acquisition System

Authors: V. G. Sangam, Balasaheb M. Patre

Abstract:

In this paper we have proposed a methodology to develop an amperometric biosensor for the analysis of glucose concentration using a simple microcontroller based data acquisition system. The work involves the development of Detachable Membrane Unit (enzyme based biomembrane) with immobilized glucose oxidase on the membrane and interfacing the same to the signal conditioning system. The current generated by the biosensor for different glucose concentrations was signal conditioned, then acquired and computed by a simple AT89C51-microcontroller. The optimum operating parameters for the better performance were found and reported. The detailed performance evaluation of the biosensor has been carried out. The proposed microcontroller based biosensor system has the sensitivity of 0.04V/g/dl, with a resolution of 50mg/dl. It has exhibited very good inter day stability observed up to 30 days. Comparing to the reference method such as HPLC, the accuracy of the proposed biosensor system is well within ± 1.5%. The system can be used for real time analysis of glucose concentration in the field such as, food and fermentation and clinical (In-Vitro) applications.

Keywords: Biosensor, DMU, Glucose oxidase andMicrocontroller.

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159 The Improved Biofuel Cell for Electrical Power Generation from Wastewaters

Authors: M. S. Kilic, S. Korkut, B. Hazer

Abstract:

Newly synthesized Polypropylene-g-Polyethylene glycol polymer was first time used for a compartment-less enzymatic fuel cell. Working electrodes based on Polypropylene-g-Polyethylene glycol were operated as unmediated and mediated system (with ferrocene and gold/cobalt oxide nanoparticles). Glucose oxidase and bilirubin oxidase was selected as anodic and cathodic enzyme, respectively. Glucose was used as fuel in a single-compartment and membrane-less cell. Maximum power density was obtained as 0.65 nW cm-2, 65 nW cm-2 and 23500 nW cm-2 from the unmediated, ferrocene and gold/cobalt oxide modified polymeric film, respectively. Power density was calculated to be ~16000 nW cm-2 for undiluted wastewater sample with gold/cobalt oxide nanoparticles including system.

Keywords: Bilirubin oxidase, Enzymatic fuel cell, Glucose oxidase, Nanoparticles.

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158 Characterization of ZrO2/PEG Composite Film as Immobilization Matrix for Glucose Oxidase

Authors: N. M. Ahmad, J. Abdullah, N. I. Ramli, S. Abd Rahman, N. E. Azmi, Z. Hamzah, A. Saat, N. H. Rahman

Abstract:

A biosensor based on glucose oxidase (GOx) immobilized onto nanoparticles zirconium oxide with polyethylene nanocomposite for glucose monitoring has been designed. The CTAB/PEG/ZrO2/GOx nanocomposite was deposited onto screen printed carbon paste (SPCE) electrode via spin coating technique. The properties of CTAB/PEG/ZrO2/GOx were study using scanning electron microscopy (SEM). The SPE modified with the CTAB/PEG/ZrO2/GOx showed electrocatalytical response to the oxidation of glucose when ferrocene carboxaldehyde was used as an artificial redox mediator, which was studied by cyclic voltammetry (CV). Several parameters such as working potential, effect of pH and effect of ZrO2/PEG layers that governed the analytical performance of the biosensor, have been studied. The biosensor was applied to detect glucose with a linear range of 0.4 to 2.0 mmol L−1 with good repetability and reproducibility.

Keywords: Nanocomposite, Nanoparticles, Modified SPE, Ferrocenecarboxaldehyde.

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157 Induction of Alternative Oxidase Activity in Candida albicans by Oxidising Conditions

Authors: Simon Brown, Raewyn Tuffery

Abstract:

Candida albicans ATCC 10231 had low endogenous activity of the alternative oxidase compared with that of C. albicans ATCC 10261. In C. albicans ATCC 10231 the endogenous activity declined as the cultures aged. Alternative oxidase activity could be induced in C. albicans ATCC 10231 by treatment with cyanide, but the induction of this activity required the presence of oxygen which could be replaced, at least in part, with high concentrations of potassium ferricyanide. We infer from this that the expression of the gene encoding the alternative oxidase is under the control of a redoxsensitive transcription factor.

Keywords: alternative oxidase, Candida albicans, enzymeinduction, oxygen, redox potential.

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156 Role of Fish Hepatic Aldehyde Oxidase in Oxidative in vitro Metabolism of Phenanthridine Heterocyclic Aromatic Compound

Authors: Khaled S. Al Salhen

Abstract:

Aldehyde oxidase is molybdo-flavoenzyme involved in the oxidation of hundreds of endogenous and exogenous and N-heterocyclic compounds and environmental pollutants. Uncharged N-heterocyclic aromatic compounds such phenanthridine are commonly distributed pollutants in soil, air, sediments, surface water and groundwater, and in animal and plant tissues. Phenanthridine as uncharged N-heterocyclic aromatic compound was incubated with partially purified aldehyde oxidase from rainbow trout fish liver. Reversed-phase HLPC method was used to separate the oxidation products from phenanthridine and the metabolite was identified. The 6(5H)-phenanthridinone was identified the major metabolite by partially purified aldehyde oxidase from fish liver. Kinetic constant for the oxidation reactions were determined spectrophotometrically and showed that this substrate has a good affinity (Km = 78 ± 7.6µM) for hepatic aldehyde oxidase, will be a significant pathway. This study confirms that partially purified aldehyde oxidase from fish liver is indeed the enzyme responsible for the in vitro production 6(5H)-phenanthridinone metabolite as it is a major metabolite by mammalian aldehyde oxidase, coupled with a relatively high oxidation rate (0.77± 0.03 nmol/min/mg protein). In addition, the kinetic parameters of hepatic fish aldehyde oxidase towards the phenanthridine substrate indicate that in vitro biotransformation by hepatic fish aldehyde oxidase will be a significant pathway. This study confirms that partially purified aldehyde oxidase from fish liver is indeed the enzyme responsible for the in vitro production 6(5H)-phenanthridinone metabolite as it is a major metabolite by mammalian aldehyde oxidase.

Keywords: Aldehyde oxidase, Fish, Phenanthridine, Specificity.

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155 Production of Glucose from the Hydrolysis of Cassava Residue using Bacteria Isolates from Thai Higher Termites

Authors: Pitcha Wongskeo, Pramoch Rangsunvigit, Sumaeth Chavadej

Abstract:

The possibility of using cassava residue containing 49.66% starch, 21.47% cellulose, 12.97% hemicellulose, and 21.86% lignin as a raw material to produce glucose using enzymatic hydrolysis was investigated. In the experiment, each reactor contained the cassava residue, bacteria cells, and production medium. The effects of particles size (40 mesh and 60 mesh) and strains of bacteria (A002 and M015) isolated from Thai higher termites, Microcerotermes sp., on the glucose concentration at 37°C were focused. High performance liquid chromatography (HPLC) with a refractive index detector was used to determine the quantity of glucose. The maximum glucose concentration obtained at 37°C using strain A002 and 60 mesh of the cassava residue was 1.51 g/L at 10 h.

Keywords: Hydrolysis, termites, glucose, cassava

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154 Glucose-dependent Functional Heterogeneity In β-TC-6 Murine Insulinoma

Authors: Darren C-W. Tan, Partha Roy

Abstract:

To determine if the murine insulinoma, β-TC-6, is a suitable substitute for primary pancreatic β-cells in the study of β- cell functional heterogeneity, we used three distinct functional assays to ascertain the cell line-s response to glucose or a glucose analog. These assays include: (i) a 2-NBDG uptake assay; (ii) a calcium influx assay, and; (iii) a quinacrine secretion assay. We show that a population of β-TC-6 cells endocytoses the glucose analog, 2- NBDG, at different rates, has non-uniform intracellular calcium ion concentrations and releases quinacrine at different rates when challenged with glucose. We also measured the Km for β-TC-6 glucose uptake to be 46.9 mM and the Vm to be 8.36 x 10-5 mmole/million cells/min. These data suggest that β-TC-6 might be used as an alternative to primary pancreatic β-cells for the study of glucose-dependent β-cell functional heterogeneity.

Keywords: 2-NBDG, Fura-2/AM, functional heterogeneity, quinacrine.

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153 A Novel Method for Blood Glucose Measurement by Noninvasive Technique Using Laser

Authors: V.Ashok, A.Nirmalkumar, N.Jeyashanthi

Abstract:

A method and apparatus for noninvasive measurement of blood glucose concentration based on transilluminated laser beam via the Index Finger has been reported in this paper. This method depends on atomic gas (He-Ne) laser operating at 632.8nm wavelength. During measurement, the index finger is inserted into the glucose sensing unit, the transilluminated optical signal is converted into an electrical signal, compared with the reference electrical signal, and the obtained difference signal is processed by signal processing unit which presents the results in the form of blood glucose concentration. This method would enable the monitoring blood glucose level of the diabetic patient continuously, safely and noninvasively.

Keywords: Anisotropy factor, Blood glucose, Diabetes Mellitus, Noninvasive method, Photo detectors.

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152 Causal Modeling of the Glucose-Insulin System in Type-I Diabetic Patients

Authors: J. Fernandez, N. Aguilar, R. Fernandez de Canete, J. C. Ramos-Diaz

Abstract:

In this paper, a simulation model of the glucose-insulin system for a patient undergoing diabetes Type 1 is developed by using a causal modeling approach under system dynamics. The OpenModelica simulation environment has been employed to build the so called causal model, while the glucose-insulin model parameters were adjusted to fit recorded mean data of a diabetic patient database. Model results under different conditions of a three-meal glucose and exogenous insulin ingestion patterns have been obtained. This simulation model can be useful to evaluate glucose-insulin performance in several circumstances, including insulin infusion algorithms in open-loop and decision support systems in closed-loop.

Keywords: Causal modeling, diabetes, glucose-insulin system, diabetes, causal modeling, OpenModelica software.

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151 Blood Glucose Level Measurement from Breath Analysis

Authors: Tayyab Hassan, Talha Rehman, Qasim Abdul Aziz, Ahmad Salman

Abstract:

The constant monitoring of blood glucose level is necessary for maintaining health of patients and to alert medical specialists to take preemptive measures before the onset of any complication as a result of diabetes. The current clinical monitoring of blood glucose uses invasive methods repeatedly which are uncomfortable and may result in infections in diabetic patients. Several attempts have been made to develop non-invasive techniques for blood glucose measurement. In this regard, the existing methods are not reliable and are less accurate. Other approaches claiming high accuracy have not been tested on extended dataset, and thus, results are not statistically significant. It is a well-known fact that acetone concentration in breath has a direct relation with blood glucose level. In this paper, we have developed the first of its kind, reliable and high accuracy breath analyzer for non-invasive blood glucose measurement. The acetone concentration in breath was measured using MQ 138 sensor in the samples collected from local hospitals in Pakistan involving one hundred patients. The blood glucose levels of these patients are determined using conventional invasive clinical method. We propose a linear regression classifier that is trained to map breath acetone level to the collected blood glucose level achieving high accuracy.

Keywords: Blood glucose level, breath acetone concentration, diabetes, linear regression.

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150 Study on the Effect of Sulphur, Glucose, Nitrogen and Plant Residues on the Immobilization of Sulphate-S in Soil

Authors: S. Shahsavani, A. Gholami

Abstract:

In order to evaluate the relationship between the sulphur (S), glucose (G), nitrogen (N) and plant residues (st), sulphur immobilization and microbial transformation were monitored in five soil samples from 0-30 cm of Bastam farmers fields of Shahrood area following 11 treatments with different levels of Sulphur (S), glucose (G), N and plant residues (wheat straw) in a randomized block design with three replications and incubated over 20, 45 and 60 days, the immobilization of SO4 -2-S presented as a percentage of that added, was inversely related to its addition rate. Additions of glucose and plant residues increased with the C-to-S ratio of the added amendments, irrespective of their origins (glucose and plant residues). In the presence of C sources (glucose or plant residues). N significantly increased the immobilization of SO4 -2-S, whilst the effect of N was insignificant in the absence of a C amendment. In first few days the amounts of added SO4 -2-S immobilized were linearly correlated with the amounts of added S recovered in the soil microbial biomass. With further incubation the proportions of immobilized SO4 -2-S remaining as biomass-S decreased. Decrease in biomass-S was thought to be due to the conversion of biomass-S into soil organic-S. Glucose addition increased the immobilization (microbial utilization and incorporation into the soil organic matter) of native soil SO4 -2-S. However, N addition enhance the mineralization of soil organic-S, increasing the concentration of SO4 - 2-S in soil.

Keywords: Immobilization, microbial biomass, sulphur, nitrogen, glucose.

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149 Effect of Nitrogen and Carbon Sources on Growth and Lipid Production from Mixotrophic Growth of Chlorella sp. KKU-S2

Authors: Ratanaporn Leesing, Thidarat Papone, Mutiyaporn Puangbut

Abstract:

Mixotrophic cultivation of the isolated freshwater microalgae Chlorella sp. KKU-S2 in batch shake flask for biomass and lipid productions, different concentration of glucose as carbon substrate, different nitrogen source and concentrations were investigated. Using 1.0g/L of NaNO3 as nitrogen source, the maximum biomass yield of 10.04g/L with biomass productivity of 1.673g/L d was obtained using 40g/L glucose, while a biomass of 7.09, 8.55 and 9.45g/L with biomass productivity of 1.182, 1.425 and 1.575g/L d were found at 20, 30 and 50g/L glucose, respectively. The maximum lipid yield of 3.99g/L with lipid productivity of 0.665g/L d was obtained when 40g/L glucose was used. Lipid yield of 1.50, 3.34 and 3.66g/L with lipid productivity of 0.250, 0.557 and 0.610g/L d were found when using the initial concentration of glucose at 20, 30 and 50g/L, respectively. Process product yield (YP/S) of 0.078, 0.119, 0.158 and 0.094 were observed when glucose concentration was 20, 30, 40 and 50 g/L, respectively. The results obtained from the study shows that mixotrophic culture of Chlorella sp. KKU-S2 is a desirable cultivation process for microbial lipid and biomass production. 

Keywords: Mixotrophic cultivation, microalgal lipid, Chlorella sp. KKU-S2.

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148 An Algorithm of Regulation of Glucose-Insulin Concentration in the Blood

Authors: B. Selma, S. Chouraqui

Abstract:

The pancreas is an elongated organ that extends across the abdomen, below the stomach. In addition, it secretes certain enzymes that aid in food digestion. The pancreas also manufactures hormones responsible for regulating blood glucose levels. In the present paper, we propose a mathematical model to study the homeostasis of glucose and insulin in healthy human, and a simulation of this model, which depicts the physiological events after a meal, will be represented in ordinary humans. The aim of this paper is to design an algorithm which regulates the level of glucose in the blood. The algorithm applied the concept of expert system for performing an algorithm control in the form of an "active" used to prescribe the rate of insulin infusion. By decomposing the system into subsystems, we have developed parametric models of each subsystem by using a forcing function strategy. The results showed a performance of the control system.

Keywords: Modeling, algorithm, regulation, glucose-insulin, blood, control system.

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147 A Modern Review of the Non-Invasive Continuous Blood Glucose Measuring Devices and Techniques for Remote Patient Monitoring System

Authors: Muhibul Haque Bhuyan

Abstract:

Diabetes disease that arises from the higher glucose level due to insulin shortage or insulin opposition in the human body has become a common disease in the world. No medicine can cure it completely. However, by taking medicine, maintaining diets, and having exercises regularly, a diabetes patient can keep his glucose level within the specified limits and in this way, he/she can lead a normal life like a healthy person. But to control glucose levels, a patient needs to monitor them regularly. Various techniques are being used over the last four decades. This modern review article aims to provide a comparative study report on various blood glucose monitoring techniques in a very concise and organized manner. The review mainly emphasizes working principles, cost, technology, sensors, measurement types, measurement accuracy, advantages, and disadvantages, etc. of various techniques and then compares among each other. Besides, the use of algorithms and simulators for the growth of this technology is also presented. Finally, current research trends of this measurement technology have also been discussed.

Keywords: blood glucose measurement, sensors, measurement devices, invasive and non-invasive techniques

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146 Improved Blood Glucose-Insulin Monitoring with Dual-Layer Predictive Control Design

Authors: Vahid Nademi

Abstract:

In response to widely used wearable medical devices equipped with a continuous glucose monitor (CGM) and insulin pump, the advanced control methods are still demanding to get the full benefit of these devices. Unlike costly clinical trials, implementing effective insulin-glucose control strategies can provide significant contributions to the patients suffering from chronic diseases such as diabetes. This study deals with a key role of two-layer insulin-glucose regulator based on model-predictive-control (MPC) scheme so that the patient’s predicted glucose profile is in compliance with the insulin level injected through insulin pump automatically. It is achieved by iterative optimization algorithm which is called an integrated perturbation analysis and sequential quadratic programming (IPA-SQP) solver for handling uncertainties due to unexpected variations in glucose-insulin values and body’s characteristics. The feasibility evaluation of the discussed control approach is also studied by means of numerical simulations of two case scenarios via measured data. The obtained results are presented to verify the superior and reliable performance of the proposed control scheme with no negative impact on patient safety.

Keywords: Blood glucose monitoring, insulin pump, optimization, predictive control, diabetes disease.

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145 Linear Prediction System in Measuring Glucose Level in Blood

Authors: Intan Maisarah Abd Rahim, Herlina Abdul Rahim, Rashidah Ghazali

Abstract:

Diabetes is a medical condition that can lead to various diseases such as stroke, heart disease, blindness and obesity. In clinical practice, the concern of the diabetic patients towards the blood glucose examination is rather alarming as some of the individual describing it as something painful with pinprick and pinch. As for some patient with high level of glucose level, pricking the fingers multiple times a day with the conventional glucose meter for close monitoring can be tiresome, time consuming and painful. With these concerns, several non-invasive techniques were used by researchers in measuring the glucose level in blood, including ultrasonic sensor implementation, multisensory systems, absorbance of transmittance, bio-impedance, voltage intensity, and thermography. This paper is discussing the application of the near-infrared (NIR) spectroscopy as a non-invasive method in measuring the glucose level and the implementation of the linear system identification model in predicting the output data for the NIR measurement. In this study, the wavelengths considered are at the 1450 nm and 1950 nm. Both of these wavelengths showed the most reliable information on the glucose presence in blood. Then, the linear Autoregressive Moving Average Exogenous model (ARMAX) model with both un-regularized and regularized methods was implemented in predicting the output result for the NIR measurement in order to investigate the practicality of the linear system in this study. However, the result showed only 50.11% accuracy obtained from the system which is far from the satisfying results that should be obtained.

Keywords: Diabetes, glucose level, linear, near-infrared (NIR), non-invasive, prediction system.

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144 Salicylhydroxamic Acid Inhibits the Growth of Candida albicans

Authors: Shu-Ying Marissa Pang, Stephen Tristram, Simon Brown

Abstract:

Candida spp. are common and aggressive pathogens. Because of the growing resistance of Candida spp. to current antifungals, novel targets, found in Candida spp. but not in humans or other flora, have to be identified. The alternative oxidase (AOX) is one such possibility. This enzyme is insensitive to cyanide, but is sensitive to compounds such as salicylhydroxamic acid (SHAM), disulfiram and n-alkyl gallates. The growth Candida albicans was inhibited by SHAM (Ki = 9-15 mM) and cyanide (Ki = 2-4 mM), albeit to differing extents. The rate of O2 uptake was inhibited by less than 10% by 25 mM SHAM and by about 90% by 250 μM KCN. Although SHAM substantially inhibited the growth of C. albicans, it is unlikely that the inhibition of AOX was the cause. Salicylhydroxamic acid is used therapeutically in the treatment of urinary tract infections and urolithiasis, but it also has some potential in the treatment of C. albicans infection.

Keywords: alternative oxidase, Candida albicans, growth, respiration, salicylhydroxamic acid.

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143 A Study of Fatty Acid Production in the Batch Reactor via the Carbohydrate Fermentation by C. butyricum

Authors: H. Azan T., R. W. Lovitt, Nur K. T., N. Azwa. M. B.

Abstract:

Carbohydrate can be used as a substrate that can be consumed by C. butyricum and converted to useful chemicals such as acetic and butyric acid. Influence of concentration and types of carbohydrate to cell growth, carbohydrate consumed, productivity and carbon balance have been explored. Batch reactor was selected in this study to avoid contamination due to simpler operation system. Glucose was preferred as first types of carbohydrate to be tested. Six concentrations were studied from 0 to 28g/L. Eventually, 15g/L has shown the best concentration for glucose in term of growth rate (2.63h-1) and carbon balance (99.76% recovery). Comparison for types of carbohydrate was also conducted. 15g/L of xylose (monosaccharide) and starch (complex carbohydrate) was tested.  In term of growth rate and productivity, glucose showed the best carbohydrates. Results for this study showed that glucose and xylose produced more than 80% of acetic acid and less than 20% of butyric acid. Meanwhile, 63.1% of acetic acid and 36.9% of butyric acid were produced from starch. 

Keywords: C. butyricum, glucose, starch, xylose, carbohydrate.

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142 Inhibition of the Growth of Pathogenic Candida spp. by Salicylhydroxamic Acid

Authors: Shu-Ying Marissa Pang, Stephen Tristram, Simon Brown

Abstract:

Candida spp. are common and aggressive pathogens. Because of the growing resistance of Candida spp. to current antifungals, novel targets, found in Candida spp. but not in humans or other flora, have to be identified. The alternative oxidase (AOX) is one such possibility. This enzyme is insensitive to cyanide, but is sensitive to compounds such as salicylhydroxamic acid (SHAM), disulfiram and n-alkyl gallates. The growth each of six Candida spp. was inhibited significantly by ~13 mM SHAM or 2 mM cyanide, albeit to differing extents. In C. dubliniensis, C. krusei and C. tropicalis the rate of O2 uptake was inhibited by 18-36% by 25 mM SHAM, but this had little or no effect on C. glabrata, C. guilliermondii or C. parapsilosis. Although SHAM substantially inhibited the growth of Candida spp., it is unlikely that the inhibition of AOX was the cause. Salicylhydroxamic acid is used therapeutically in the treatment of urinary tract infections and urolithiasis, but it also has some potential in the treatment of Candida spp. infection.

Keywords: alternative oxidase, Candida spp., growth, respiration, salicylhydroxamic acid.

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141 Blood Glucose Measurement and Analysis: Methodology

Authors: I. M. Abd Rahim, H. Abdul Rahim, R. Ghazali

Abstract:

There is numerous non-invasive blood glucose measurement technique developed by researchers, and near infrared (NIR) is the potential technique nowadays. However, there are some disagreements on the optimal wavelength range that is suitable to be used as the reference of the glucose substance in the blood. This paper focuses on the experimental data collection technique and also the analysis method used to analyze the data gained from the experiment. The selection of suitable linear and non-linear model structure is essential in prediction system, as the system developed need to be conceivably accurate.

Keywords: Invasive, linear, near-infrared (Nir), non-invasive, non-linear, prediction system.

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140 A Neural Network Approach in Predicting the Blood Glucose Level for Diabetic Patients

Authors: Zarita Zainuddin, Ong Pauline, C. Ardil

Abstract:

Diabetes Mellitus is a chronic metabolic disorder, where the improper management of the blood glucose level in the diabetic patients will lead to the risk of heart attack, kidney disease and renal failure. This paper attempts to enhance the diagnostic accuracy of the advancing blood glucose levels of the diabetic patients, by combining principal component analysis and wavelet neural network. The proposed system makes separate blood glucose prediction in the morning, afternoon, evening and night intervals, using dataset from one patient covering a period of 77 days. Comparisons of the diagnostic accuracy with other neural network models, which use the same dataset are made. The comparison results showed overall improved accuracy, which indicates the effectiveness of this proposed system.

Keywords: Diabetes Mellitus, principal component analysis, time-series, wavelet neural network.

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139 Microfluidic Paper-Based Electrochemical Biosensor

Authors: Ahmad Manbohi, Seyyed Hamid Ahmadi

Abstract:

A low-cost paper-based microfluidic device (PAD) for the multiplex electrochemical determination of glucose, uric acid, and dopamine in biological fluids was developed. Using wax printing, PAD containing a central zone, six channels, and six detection zones was fabricated, and the electrodes were printed on detection zones using pre-made electrodes template. For each analyte, two detection zones were used. The carbon working electrode was coated with chitosan-BSA (and enzymes for glucose and uric acid). To detect glucose and uric acid, enzymatic reactions were employed. These reactions involve enzyme-catalyzed redox reactions of the analytes and produce free electrons for electrochemical measurement. Calibration curves were linear (R² > 0.980) in the range of 0-80 mM for glucose, 0.09–0.9 mM for dopamine, and 0–50 mM for uric acid, respectively. Blood samples were successfully analyzed by the proposed method.

Keywords: Multiplex, microfluidic paper-based electrochemical biosensors, biomarkers, biological fluids.

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138 Investigations of Metals and Metal-Antibrowning Agents Effects on Polyphenol Oxidase Activity from Red Poppy Leaf

Authors: G. Arabaci

Abstract:

Heavy metals are one of the major groups of contaminants in the environment and many of them are toxic even at very low concentration in plants and animals. However, some metals play important roles in the biological function of many enzymes in living organisms. Metals such as zinc, iron, and cooper are important for survival and activity of enzymes in plants, however heavy metals can inhibit enzyme which is responsible for defense system of plants. Polyphenol oxidase (PPO) is a copper-containing metalloenzyme which is responsible for enzymatic browning reaction of plants. Enzymatic browning is a major problem for the handling of vegetables and fruits in food industry. It can be increased and effected with many different futures such as metals in the nature and ground. In the present work, PPO was isolated and characterized from green leaves of red poppy plant (Papaverr hoeas). Then, the effect of some known antibrowning agents which can form complexes with metals and metals were investigated on the red poppy PPO activity. The results showed that glutathione was the most potent inhibitory effect on PPO activity. Cu(II) and Fe(II) metals increased the enzyme activities however, Sn(II) had the maximum inhibitory effect and Zn(II) and Pb(II) had no significant effect on the enzyme activity. In order to reduce the effect of heavy metals, the effects of metal-antibrowning agent complexes on the PPO activity were determined. EDTA and metal complexes had no significant effect on the enzyme. L-ascorbic acid and metal complexes decreased but L-ascorbic acid-Cu(II)-complex had no effect. Glutathione–metal complexes had the best inhibitory effect on Red poppy leaf PPO activity.

Keywords: Inhibition, metal, red poppy, Polyphenol oxidase (PPO).

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137 High Glucose Increases Acetylcholine-Induced Ca2+ Entry and Protein Expression of STIM1

Authors: Hong Ding, Fatiha Benslimane, Isra Marei, Chris R. Triggle

Abstract:

Hyperglycaemia is a key factor that contributes to the development of diabetes-related microvascular disease and a major risk factor for endothelial dysfunction. In the current study, we have explored glucose-induced abnormal intracellular calcium (Ca2+ i) homeostasis in mouse microvessel endothelial cells (MMECs) in high glucose (HG) (40mmol/L) versus control (low glucose, LG) (11 mmol/L). We demonstrated that the exposure of MMECs to HG for 3 days did not change basal Ca2+ i, however, there was a significant increase of acetylcholine-induced Ca2+ entry. Western blots illustrated that exposure to HG also increased STIM1 (Stromal Interaction Molecule 1), but not Orai1 (the pore forming subunit), protein expression levels. Although the link between HG-induced changes in STIM1 expression, enhanced Ca2+ entry and endothelial dysfunction requires further study, the current data are suggestive that targeting these pathways may reduce the impact of HG on endothelial function.

Keywords: store-operated calcium entry, hyperglycaemia, STIM1, endothelial dysfunction

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136 Metabolic Analysis of Fibroblast Conditioned Media and Comparison with Theoretical Modeling

Authors: Priyanka Gupta, Paul Verma, Kerry Hourigan, Jayesh Bellare, Sameer Jadhav

Abstract:

Understanding the consumption and production of various metabolites of fibroblast conditioned media is needed for its proper and optimized use in expansion of pluripotent stem cells. For this purpose, we have used the HPLC method to analyse the consumption of glucose and the production of lactate over time by mouse embryonic fibroblasts. The experimental data have also been compared with mathematical model fits. 0.025 moles of lactate was produced after 72 hrs while the glucose concentration decreased from 0.017 moles to 0.011 moles. The mathematical model was able to predict the trends of glucose consumption and lactate production.

Keywords: Conditioned media, HPLC, metabolite analysis, mouse embryonic fibroblast.

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135 The Efficiency of Cytochrome Oxidase Subunit 1 Gene (cox1) in Reconstruction of Phylogenetic Relations among Some Crustacean Species

Authors: Yasser M. Saad, Heba El-Sebaie Abd El-Sadek

Abstract:

Some Metapenaeus monoceros cox1 gene fragments were isolated, purified, sequenced, and comparatively analyzed with some other Crustacean Cox1 gene sequences (obtained from National Center for Biotechnology Information). This work was designed for testing the efficiency of this system in reconstruction of phylogenetic relations among some Crustacean species belonging to four genera (Metapenaeus, Artemia, Daphnia and Calanus). The single nucleotide polymorphism and haplotype diversity were calculated for all estimated mt-DNA fragments. The genetic distance values were 0.292, 0.015, 0.151, and 0.09 within Metapenaeus species, Calanus species, Artemia species, and Daphnia species, respectively. The reconstructed phylogenetic tree is clustered into some unique clades. Cytochrome oxidase subunit 1 gene (cox1) was a powerful system in reconstruction of phylogenetic relations among evaluated crustacean species.

Keywords: Crustacean, Genetics, cox1, phylogeny.

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134 Fermentation of Xylose and Glucose Mixture in Intensified Reactors by Scheffersomyces stipitis to Produce Ethanol

Authors: S. C. Santos, S. R. Dionísio, A. L. D. De Andrade, L. R. Roque, A. C. Da Costa, J. L. Ienczak

Abstract:

In this work, two fermentations at different temperatures (25 and 30ºC), with cell recycling, were accomplished to produce ethanol, using a mix of commercial substrates, xylose (70%) and glucose (30%), as organic source for Scheffersomyces stipitis. Five consecutive fermentations of 80 g L-1 (1º, 2º and 3º recycles), 96 g L-1 (4º recycle) and 120 g L-1 (5º recycle)reduced sugars led to a final maximum ethanol concentration of 17.2 and 34.5 g L-1, at 25 and 30ºC, respectively. Glucose was the preferred substrate; moreover xylose startup degradation was initiated after a remaining glucose presence in the medium. Results showed that yeast acid treatment, performed before each cycle, provided improvements on cell viability, accompanied by ethanol productivity of 2.16 g L-1 h- 1 at 30ºC. A maximum 36% of xylose was retained in the fermentation medium and after five-cycle fermentation an ethanol yield of 0.43 g ethanol/g sugars was observed. S. stipitis fermentation capacity and tolerance showed better results at 30ºC with 83.4% of theoretical yield referenced on initial biomass.

Keywords: 5-carbon sugar, cell recycling fermenter, mixed sugars, xylose-fermenting yeast.

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133 Hypoglycemic Activity of Water Soluble Polysaccharides of Yam (Dioscorea hispida Dents) Prepared by Aqueous, Papain, and Tempeh Inoculum Assisted Extractions

Authors: Teti Estiasih, Harijono, Weny Bekti Sunarharum, Atina Rahmawati

Abstract:

This research studied the hypoglycemic effect of water soluble polysaccharide (WSP) extracted from yam (Dioscorea hispida) tuber by three different methods: aqueous extraction, papain assisted extraction, and tempeh inoculums assisted extraction. The two later extraction methods were aimed to remove WSP binding protein to have more pure WSP. The hypoglycemic activities were evaluated by means in vivo test on alloxan induced hyperglycemic rats, glucose response test (GRT), in situ glucose absorption test using everted sac, and short chain fatty acids (SCFAs) analysis. All yam WSP extracts exhibited ability to decrease blood glucose level in hyperglycemia condition as well as inhibited glucose absorption and SCFA formation. The order of hypoglycemic activity was tempeh inoculums assisted- >papain assisted- >aqueous WSP extracts. GRT and in situ glucose absorption test showed that order of inhibition was papain assisted- >tempeh inoculums assisted- >aqueous WSP extracts. Digesta of caecum of yam WSP extracts oral fed rats had more SCFA than control. Tempeh inoculums assisted WSP extract exhibited the most significant hypoglycemic activity.

Keywords: hypoglycemic activity, papain, tempeh inoculums, water soluble polysaccharides, yam (Discorea hispida)

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132 Extractive Fermentation of Ethanol Using Vacuum Fractionation Technique

Authors: Weeraya Samnuknit, Apichat Boontawan

Abstract:

A vacuum fractionation technique was introduced to remove ethanol from fermentation broth. The effect of initial glucose and ethanol concentrations were investigated for specific productivity. The inhibitory ethanol concentration was observed at 100 g/L. In order to increase the fermentation performance, the ethanol product was removed as soon as it is produced. The broth was boiled at 35oC by reducing the pressure to 65 mBar. The ethanol/water vapor was fractionated for up to 90 wt% before leaving the column. Ethanol concentration in the broth was kept lower than 25 g/L, thus minimized the product inhibition effect to the yeast cells. For batch extractive fermentation, a high substrate utilization rate was obtained at 26.6 g/L.h and most of glucose was consumed within 21 h. For repeated-batch extractive fermentation, addition of glucose was carried out up to 9 times and ethanol was produced more than 8-fold higher than batch fermentation.

Keywords: Ethanol, Extractive fermentation, Product inhibition, Vacuum fractionation.

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131 Molecular Characterization of Echinococcus granulosus through Amplification of 12S rRNA Gene and Cox1 Gene Fragments from Cattle in Chittagong, Bangladesh

Authors: M. Omer Faruk, A. M. A. M. Zonaed Siddiki, M. Fazal Karim, Md. Masuduzzaman, S. Chowdhury, Md. Shafiqul Islam, M. Alamgir Hossain

Abstract:

The dog tapeworms Echinococcus granulosus develop hydatid cysts in various organs in human and domestic animals worldwide including Bangladesh. The aim of this study was to identify and characterize the genotype of E. granulosus isolated from cattle using 12S rRNA and Cytochrome oxidase 1 (COX 1) genes. A total of 43 hydatid cyst samples were collected from 390 examined cattle samples derived from slaughterhouses. Among them, three cysts were fertile. Genomic DNA was extracted from germinal membrane and/or protoscoleces followed by PCR amplification of mitochondrial 12S rRNA and Cytochrome oxidase 1 gene fragments. The sequence data revealed existence of G1 (64.28%) and possible G3 (21.43%) genotypes for the first time in Bangladesh. The study indicates that common sheep strain G1 is the dominant subtype of E. granulosus in Chittagong region of Bangladesh. This will increase our understanding of the epidemiology of hydatidosis in the southern part of the country and will be useful to plan suitable control measures in the long run.

Keywords: Echinococcus granulosus, molecular characterization, cattle, Bangladesh.

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