Characterization of Silica Nanoparticles in Interaction with Escherichia coli Bacteria

The objective of the present investigation was to evaluate the morphology of Escherchia coli bacteria in interaction with SiO2 nanoparticles. This study was made by atomic force microscopy and quartz crystal microbalance using SiO2 nanoparticles with 10nm, 50nm and 100nm diameter and bacteria immobilized on polyelectrolyte multilayer films obtained by spin coating or by “layer by layer” (LbL) method.

Development of an Immunoassay Platform for Diagnosis of Acute Kidney Injury

Acute kidney injury (AKI) is a new worldwide public health problem. A diagnosis of this disease using creatinine is still a problem in clinical practice. Therefore, a measurement of biomarkers responsible for AKI has received much attention in the past couple years. Cytokine interleukin-18 (IL-18) was reported as one of the early biomarkers for AKI. The most commonly used method to detect this biomarker is an immunoassay. This study used a planar platform to perform an immunoassay using fluorescence for detection. In this study, anti-IL-18 antibody was immobilized onto a microscope slide using a covalent binding method. Make-up samples were diluted at the concentration between 10 to 1000 pg/ml to create a calibration curve. The precision of the system was determined using a coefficient of variability (CV), which was found to be less than 10%. The performance of this immunoassay system was compared with the measurement from ELISA.

The Study of the Interaction between Catanionic Surface Micelle SDS-CTAB and Insulin at Air/Water Interface

Herein, we report the different types of surface morphology due to the interaction between the pure protein Insulin (INS) and catanionic surfactant mixture of Sodium Dodecyl Sulfate (SDS) and Cetyl Trimethyl Ammonium Bromide (CTAB) at air/water interface obtained by the Langmuir-Blodgett (LB) technique. We characterized the aggregations by Scanning Electron Microscopy (SEM), Atomic Force Microscopy (AFM) and Fourier transform infrared spectroscopy (FTIR) in LB films. We found that the INS adsorption increased in presence of catanionic surfactant at air/water interface. The presence of small amount of surfactant induces two-stage growth kinetics due to the pure protein absorption and protein-catanionic surface micelle interaction. The protein remains in native state in presence of small amount of surfactant mixture. Smaller amount of surfactant mixture with INS is producing surface micelle type structure. This may be considered for drug delivery system. On the other hand, INS becomes unfolded and fibrillated in presence of higher amount of surfactant mixture. In both the cases, the protein was successfully immobilized on a glass substrate by the LB technique. These results may find applications in the fundamental science of the physical chemistry of surfactant systems, as well as in the preparation of drug-delivery system.

Colorectal Cancer Screening by a CEACAM-6 Immunosensor

The biomarker for colorectal cancer (CRC) is CEACAM-6 antigen (C6AG). Therefore, this study aims to develop a novel, simple and low-cost CEACAM-6 antigen immumosensor (C6AG-IMS), based on electrical impedance measurement, for precise determination of C6AG. A low-cost screen-printed graphite electrode was constructed and used as the sensor, with CEACAM-6 antibody (C6AB) immobilized on it. The procedures of sensor fabrication and antibody immobilization are simple and low-cost. Measurement of the electrical impedance at a definite frequency ranges (0.43 – 1.26 MHz) showed that the C6AG-IMS has an excellent linear (r2>0.9) response range (8.125 – 65 pg/mL), covering the normal physiological and pathological ranges of blood C6AG levels. Also, the C6AG-IMS has excellent reliability and validity, with the intraclass correlation coefficient being 0.97. In conclusion, a novel, simple, low-cost and reliable C6AG-IMS was designed and developed, being able to accurately determine blood C6AG levels in the range of pathological and normal physiological regions. The C6AG-IMS can provide a point-of-care and immediate screening results to the user at home.

Titania and Cu-Titania Composite Layer on Graphite Substrate as Negative Electrode for Li-Ion Battery

This research study the application of the immobilized TiO2 layer and Cu-TiO2 layer on graphite substrate as a negative electrode or anode for Li-ion battery. The titania layer was produced through chemical bath deposition method, meanwhile Cu particles were deposited electrochemically. A material can be used as an electrode as it has capability to intercalates Li ions into its crystal structure. The Li intercalation into TiO2/Graphite and Cu- TiO2/Graphite were analyzed from the changes of its XRD pattern after it was used as electrode during discharging process. The XRD patterns were refined by Le Bail method in order to determine the crystal structure of the prepared materials. A specific capacity and the cycle ability measurement were carried out to study the performance of the prepared materials as negative electrode of the Li-ion battery. The specific capacity was measured during discharging process from fully charged until the cut off voltage. A 300 was used as a load. The result shows that the specific capacity of Li-ion battery with TiO2/Graphite as negative electrode is 230.87 ± 1.70mAh.g-1 which is higher than the specific capacity of Li-ion battery with pure graphite as negative electrode, i.e 140.75 ±0.46mAh.g-1. Meanwhile deposition of Cu onto TiO2 layer does not increase the specific capacity, and the value even lower than the battery with TiO2/Graphite as electrode. The cycle ability of the prepared battery is only two cycles, due to the Li ribbon which was used as cathode became fragile and easily broken.

Rapid Determination of Biochemical Oxygen Demand

Biochemical Oxygen Demand (BOD) is a measure of the oxygen used in bacteria mediated oxidation of organic substances in water and wastewater. Theoretically an infinite time is required for complete biochemical oxidation of organic matter, but the measurement is made over 5-days at 20 0C or 3-days at 27 0C test period with or without dilution. Researchers have worked to further reduce the time of measurement. The objective of this paper is to review advancement made in BOD measurement primarily to minimize the time and negate the measurement difficulties. Survey of literature review in four such techniques namely BOD-BARTTM, Biosensors, Ferricyanidemediated approach, luminous bacterial immobilized chip method. Basic principle, method of determination, data validation and their advantage and disadvantages have been incorporated of each of the methods. In the BOD-BARTTM method the time lag is calculated for the system to change from oxidative to reductive state. BIOSENSORS are the biological sensing element with a transducer which produces a signal proportional to the analyte concentration. Microbial species has its metabolic deficiencies. Co-immobilization of bacteria using sol-gel biosensor increases the range of substrate. In ferricyanidemediated approach, ferricyanide has been used as e-acceptor instead of oxygen. In Luminous bacterial cells-immobilized chip method, bacterial bioluminescence which is caused by lux genes was observed. Physiological responses is measured and correlated to BOD due to reduction or emission. There is a scope to further probe into the rapid estimation of BOD.

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

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.

Optimization of Enzymatic Hydrolysis of Manihot Esculenta Root Starch by Immobilizeda-Amylase Using Response Surface Methodology

Enzymatic hydrolysis of starch from natural sources finds potential application in commercial production of alcoholic beverage and bioethanol. In this study the effect of starch concentration, temperature, time and enzyme concentration were studied and optimized for hydrolysis of cassava (Manihot esculenta) starch powder (of mesh 80/120) into glucose syrup by immobilized (using Polyacrylamide gel) a-amylase using central composite design. The experimental result on enzymatic hydrolysis of cassava starch was subjected to multiple linear regression analysis using MINITAB 14 software. Positive linear effect of starch concentration, enzyme concentration and time was observed on hydrolysis of cassava starch by a-amylase. The statistical significance of the model was validated by F-test for analysis of variance (p < 0.01). The optimum value of starch concentration temperature, time and enzyme concentration were found to be 4.5% (w/v), 45oC, 150 min, and 1% (w/v) enzyme. The maximum glucose yield at optimum condition was 5.17 mg/mL.

Bioconversion of Biodiesel Derived Crude Glycerol by Immobilized Clostridium pasteurianum: Effect of Temperature

Batch fermentation of 5, 10 and 25 g/L biodiesel derived crude glycerol was carried out at 30, 37 and 450C by Clostridium pasteurianum cells immobilized on silica. Maximum yield of 1,3-propanediol (PDO) (0.60 mol/mol), and ethanol (0.26 mol/mol) were obtained from 10 g/L crude glycerol at 30 and 370C respectively. Maximum yield of butanol (0.28 mol/mol substrate added) was obtained at 370C with 25 g/L substrate. None of the three products were detected at 45oC even after 10 days of fermentation. Only traces of ethanol (0.01 mol/mol) were detected at 450C with 5 g/L substrate. The results obtained for 25 g/L substrate utilization were fitted in first order rate equation to obtain the values of rate constant at three different temperatures for bioconversion of glycerol. First order rate constants for bioconversion of glycerol at 30, 37 and 45oC were found to be 0.198, 0.294 and 0.029/day respectively. Activation energy (Ea) for crude glycerol bioconversion was calculated to be 57.62 kcal/mol.

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

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.

Silver Modified TiO2/Halloysite Thin Films for Decontamination of Target Pollutants

 Sol-gel method has been used to fabricate nanocomposite films on glass substrates composed halloysite clay mineral and nanocrystalline TiO2. The methodology for the synthesis involves a simple chemistry method utilized nonionic surfactant molecule as pore directing agent along with the acetic acid-based solgel route with the absence of water molecules. The thermal treatment of composite films at 450oC ensures elimination of organic material and lead to the formation of TiO2 nanoparticles onto the surface of the halloysite nanotubes. Microscopy techniques and porosimetry methods used in order to delineate the structural characteristics of the materials. The nanocomposite films produced have no cracks and active anatase crystal phase with small crystallite size were deposited on halloysite nanotubes. The photocatalytic properties for the new materials were examined for the decomposition of the Basic Blue 41 azo dye in solution. These, nanotechnology based composite films show high efficiency for dye’s discoloration in spite of different halloysite quantities and small amount of halloysite/TiO2 catalyst immobilized onto glass substrates. Moreover, we examined the modification of the halloysite/TiO2 films with silver particles in order to improve the photocatalytic properties of the films. Indeed, the presence of silver nanoparticles enhances the discoloration rate of the Basic Blue 41 compared to the efficiencies obtained for unmodified films.

Biosensor Measurement of Urea Coonncentration in Human Blood Serum

An application of the highly biosensor based on pH-sensitive field immobilized urease for urea analysis was demo The main analytical characteristics of the bios determined; the conditions of urea measureme blood were optimized. A conceptual possibility biosensor for detection of urea concentratio patients suffering from renal insufficiency was sensitive and selective effect transistor and monstrated in this work. iosensor developed were ment in real samples of ility of application of the tion in blood serum of as shown.

Improvement of Lipase Catalytic Properties by Immobilization in Hybrid Matrices

Lipases are enzymes particularly amenable for immobilization by entrapment methods, as they can work equally well in aqueous or non-conventional media and long-time stability of enzyme activity and enantioselectivity is needed to elaborate more efficient bioprocesses. The improvement of Pseudomonas fluorescens (Amano AK) lipase characteristics was investigated by optimizing the immobilization procedure in hybrid organic-inorganic matrices using ionic liquids as additives. Ionic liquids containing a more hydrophobic alkyl group in the cationic moiety are beneficial for the activity of immobilized lipase. Silanes with alkyl- or aryl nonhydrolizable groups used as precursors in combination with tetramethoxysilane could generate composites with higher enantioselectivity compared to the native enzyme in acylation reactions of secondary alcohols. The optimal effect on both activity and enantioselectivity was achieved for the composite made from octyltrimethoxysilane and tetramethoxysilane at 1:1 molar ratio (60% increase of total activity following immobilization and enantiomeric ratio of 30). Ionic liquids also demonstrated valuable properties as reaction media for the studied reactions, comparable with the usual organic solvent, hexane.

Comparative Study on Production of Fructooligosaccharides by p. Simplicissimum Using Immobilized Cells and Conventional Reactor System

Fructooligosaccharides derived from microbial enzyme especially from fungal sources has been received particular attention due to its beneficial effects as prebiotics and mass production. However, fungal fermentation is always cumbersome due to its broth rheology problem that will eventually affect the production of FOS. This study investigated the efficiency of immobilized cell system using rotating fibrous bed bioreactor (RFBB) in producing fructooligosaccharides (FOS). A comparative picture with respect to conventional stirred tank bioreactor (CSTB) and RFBB has been presented. To demonstrate the effect of agitation intensity and aeration rate, a laboratory-scale bioreactor 2.5 L was operated in three phases (high, medium, low) for 48 hours. Agitation speed has a great influence on P. simplicissimum fermentation for FOS production, where the volumetric FOS productivity using RFBB is increased with almost 4 fold compared to the FOS productivity in CSTB that only 0.319 g/L/h. Rate of FOS production increased up to 1.2 fold when immobilized cells system was employed at aeration rate similar to the freely suspended cells at 2.0 vvm.

Performance of Bio-Composite Carbonized Materials in Probiotic Applications

A new composite sorbent based on carbonized rice husk (CRH) and immobilized on it living cells and inactivated cultural liquid containing antimicrobials metabolites of Bacillus subtilis CK-245 is developed. The sorption and antimicrobic activity of CRH concerning five species of Enterobacteriaceae is studied. Prospects of use of developed sorbent in medicine and veterinary science is shown.