Suna Timur

Inhibition of alpha-glucosidase by aqueous extracts of some potent antidiabetic medicinal herbs.

Abstract Diabetes mellitus is one of the most prevalant diseases of adults. Agents with α‐glucosidase inhibitory activity have been useful as oral hypoglycemic drugs for the control of hyperglycemia in patients with type 2; noninsulin‐dependent, diabetes mellitus (NIDDM). Investigation of some medicinal herbs: Urtica dioica, Taraxacum officinale, Viscum album, and Myrtus communis with α‐glucosidase inhibitor activity was conducted to identify a prophylactic effect for diabetes in vitro. All plants showed differing potent α‐glucosidase inhibitory activity. However, Myrtus communis strongly inhibited the enzyme (IC50 = 38 µg/mL). The inhibitory effect of these plants and some common antidiabetic drugs against the enzyme source (bakers yeast, rabbit liver, and small intestine) were also searched. Approximately all inhibitors used in this study showed quite different inhibitory activities, according to α‐glucosidase origins. Furthermore, subsequent separation of the active material from Myrtus communis by HPLC showed that only one fraction acted as an α‐glucosidase inhibitor.

Examination of performance of glassy carbon paste electrode modified with gold nanoparticle and xanthine oxidase for xanthine and hypoxanthine detection

A composite electrode was prepared by modifying glassy carbon microparticles with gold nanoparticles (Au-nps) and xanthine oxidase enzyme (XOD) for xanthine (X) and hypoxanthine (Hx) detection. After the optimization of the system for X, the biosensor was characterized for X and Hx. A linearity was obtained in the concentration range between 5.00 x 10(-7) and 1.00 x 10(-5) M for X with equation of y=0.24 x + 0.712 and 5.00 x 10(-6) to 1.50 x 10(-4) M for Hx, with equation of y = 0.014 x + 0.575, respectively. Obtained results were compared to X and/or Hx biosensors including/not including Au-np in the structure. The developed system was also applied for detection of Hx in canned tuna fish sample and very promising results were obtained.

Detection of phenolic compounds by thick film sensors based on Pseudomonas putida.

Amperometric biosensors using bacterial cells were developed for the determination of phenolic compounds and the measurement was based on the respiratory activity of the cells. For this purpose, Pseudomonas putida DSM 50026 which is one of the well-known phenol degrading organisms, was used as a biological component. The cells were grown in the presence of phenol as the sole source of organic carbon. As well as phenol adapted cells, the bacterium which used the glucose as the major carbon source, was also used to obtain another type of biosensor for the comparison of the responses and specificities towards different xenobiotics. The commercial oxygen electrode was used as a transducer to test the sensor responses for both induced and non-induced cells. Our results showed that the adaptation step enable us to obtain biosensor devices with different substrate specificity. Moreover, P. putida was immobilized on the surface of thick film working electrodes made of gold by using gelatin membrane cross-linked with glutaraldehyde. The biosensors were calibrated for different phenolic substances. Furthermore, phenol detection was performed in synthetic wastewater samples.

Biosensing approach for glutathione detection using glutathione reductase and sulfhydryl oxidase bienzymatic system

Chitosan membrane with glutathione reductase and sulfhydryl oxidase (SOX) was subsequently integrated onto the surface of spectrographic graphite rods for obtaining a glutathione biosensor. The working principle was based on the monitoring of O(2) consumption that correlates the concentration of glutathione during the enzymatic reaction. A linear relationship between sensor response and concentration was obtained between 0.5 and 2.0 mM for oxidized glutathione (GSSG), and 0.2-1.0 mM for reduced glutathione (GSH) in the presence of 2 microM nicotinamide adenine dinucleotide phosphate (NADPH) under the optimum working conditions. Also, reduced/oxidized glutathione were separated by HPLC and utility of bienzymatic system was investigated as an electrochemical detector for the analysis of these compounds. All data were given as a comparison of two systems: biosensor and diode array detector (DAD).

A microbial biosensor based on bacterial cells immobilized on chitosan matrix

A bio-electrochemical system consisting of Gluconobacter oxydans DSM 2343 cells as a biological material and carbon nanotube (CNT)-free and CNT-modified chitosan as immobilizing matrices has been developed. The measurement was based on the respiratory activity of the cells estimated by the oxygen consumption at -0.7 V (versus the Ag|AgCl reference electrode) due to the metabolic activity in the presence of substrates. The system was calibrated and dependence of signal amplitude on the measuring conditions and cell amount was studied as well as the substrate specificity, pH, temperature and working potential. The biosensors (CNT-modified and unmodified) were demonstrated for the quantification of glucose in the range of 0.05-1.0 mM, at 30 degrees C and pH 7.0 with the 40 s of response time. The linear relationships between sensor response (y; microA/cm(2)) and substrate concentration (x; mM) were defined by the equations of y=1.160x+0.151 (R(2)=0.990) and y=1.261x+0.197 (R(2)=0.982), respectively. All other data were also given as comparison of two systems one with CNT-modified and CNT-free.

Nanostructured Amphiphilic Star-Hyperbranched Block Copolymers for Drug Delivery

A robust drug delivery system based on nanosized amphiphilic star-hyperbranched block copolymer, namely, poly(methyl methacrylate-block-poly(hydroxylethyl methacrylate) (PMMA-b-PHEMA) is described. PMMA-b-PHEMA was prepared by sequential visible light induced self-condensing vinyl polymerization (SCVP) and conventional vinyl polymerization. All of the synthesis and characterization details of the conjugates are reported. To accomplish tumor cell targeting property, initially cell-targeting (arginylglycylaspactic acid; RGD) and penetrating peptides (Cys-TAT) were binding to each other via the well-known EDC/NHS chemistry. Then, the resulting peptide was further incorporated to the surface of the amphiphilic hyperbranched copolymer via a coupling reaction between the thiol (-SH) group of the peptide and the hydroxyl group of copolymer by using N-(p-maleinimidophenyl) isocyanate as a heterolinker. The drug release property and targeting effect of the anticancer drug (doxorobucin; DOX) loaded nanostructures to two different cell lines were evaluated in vitro. U87 and MCF-7 were chosen as integrin αvβ3 receptor positive and negative cells for the comparison of the targeting efficiency, respectively. The data showed that drug-loaded copolymers exhibited enhanced cell inhibition toward U87 cells in compared to MCF-7 cells because targeting increased the cytotoxicity of drug-loaded copolymers against integrin αvβ3 receptor expressing tumor cells.

Polypeptide Functional Surface for the Aptamer Immobilization: Electrochemical Cocaine Biosensing

Electroanalytical technologies as a beneficial subject of modern analytical chemistry can play an important role for abused drug analysis which is crucial for both legal and social respects. This article reports a novel aptamer-based biosensing procedure for cocaine analysis by combining the advantages of aptamers as selective recognition elements with the well-known advantages of biosensor systems such as the possibility of miniaturization and automation, easy fabrication and modification, low cost, and sensitivity. In order to construct the aptasensor platform, first, polythiophene bearing polyalanine homopeptide side chains (PT-Pala) was electrochemically coated onto the surface of an electrode and then cocaine aptamer was attached to the polymer via covalent conjugation chemistry. The stepwise modification of the surface was confirmed by electrochemical characterization. The designed biosensing system was applied for the detection of cocaine and its metabolite, benzoylecgonine (BE), which exhibited a linear correlation in the range from 2.5 up to 10 nM and 0.5 up to 50 μM for cocaine and BE, respectively. In order to expand its practical application, the proposed method was successfully tested for the analysis of synthetic biological fluids.

Chitosan-ferrocene film as a platform for flow injection analysis applications of glucose oxidase and Gluconobacter oxydans biosensors

Chitosan-ferrocene (CHIT-Fc) hybrid was synthesized through covalent modification and its electrochemical properties in immobilized form were studied by using cyclic voltammetry. The hybrid film exhibited reversible electrochemistry with a formal potential of +0.35 V (vs. Ag/AgCl) at pH 5.5. The Fc in CHIT matrix retained its electrocatalytic activity and did not diffuse from the matrix. This redox-active hybrid was further employed as a support for immobilization of glucose oxidase (GOx) and whole cells of Gluconobacter oxydans using glutaraldehyde on a glassy carbon electrode (GCE). The experimental conditions were optimized and the analytical characteristics of enzyme and microbial biosensors were evaluated for glucose in flow injection analysis (FIA) system. Under optimized conditions, both enzyme and microbial biosensors exhibited wide linear ranges for glucose from 2.0 to 16.0 mM and from 1.5 to 25.0 mM, respectively. Moreover, the biosensors have the advantages of relatively fast response times, good reproducibility and stability in FI mode. It was demonstrated that CHIT-Fc provides a biocompatible microenvironment for both bioctalysts and an electron transfer pathway. Additionally, integration of the enzyme and microbial biosensors into the FIA system has several advantages including capability of automation and high throughput at low cost. This promising redox hybrid can be utilized as an immobilization matrix for biomolecules in biosensor systems.

Determination of phenolic acids using Trametes versicolor laccase

Two biosensors based on Trametes versicolor laccase (TvL) were developed for the determination of phenolic compounds. Commercial oxygen electrode and ferrocene-modified screen-printed graphite electrodes were used for preparation of laccase biosensors. The systems were calibrated for three phenolic acids. Linearity was obtained in the concentration range 0.1-1.0muM caffeic acid, 0.05-0.2muM ferulic acid, 2.0-14.0muM syringic acid for laccase immobilised on a commercial oxygen electrode and 2.0-30.0muM caffeic acid, 2.0-10.0muM ferulic acid, 4.0-30.0muM syringic acid for laccase immobilised on ferrocene-modified screen-printed electrodes. Furthermore, optimal pH, temperature and thermal stability studies were performed with the commercial oxygen electrode. Both electrodes were used for determination of a class of phenolic acids, achieving a cheap and fast tool and an easy to be used procedure for screening real samples of human plasma.

PAMAM-functionalized water soluble quantum dots for cancer cell targeting

Herein, the phase-transfer reaction of quantum dots (QDs) with amine-terminated polyamidoamine (PAMAM) dendrimers with controllable ligand molar ratios was achieved. The unique properties of PAMAM allowed us to build up structurally and electrostatically stabilized water soluble QD complexes. Synthesized conjugates were characterized in terms of fluorescence and UV-Vis profiles, hydrodynamic size, number of surface dendrimer groups, and stability. Cytotoxic effects of conjugates for MCF-7, A-549 and HEP-G2 cancer cells were assessed based on cell viability using MTT assay. Cytotoxicity results were expressed as no observable adverse effect concentration (NOAEC), 50% inhibitory concentration (IC50) and total lethal concentration (TLC) values (μM). Furthermore, HER2 receptor-mediated targeting efficiency of antibody labelled P/QDs conjugates was evaluated by successful staining of MCF-7 cells with bioconjugates. Uniquely, effective cell internalization was achieved with well-characterized antibody coupled P/QDs in contrast to antibody free P/QDs conjugates. Fluorescence microscopy images demonstrated that the designed PAMAM-derivatized QDs nanoparticles show great potential in the areas of cellular imaging and targeted therapy.