Structure-activity relationship of diameter controlled Ag@Cu nanoparticles in broad-spectrum antibacterial mechanism.

Current outbreaks associated with drug-resistant clinical strains are demanding for the development of broad-spectrum antibacterial agents. The bactericidal materials should be eco-friendly, economical and effective to suppress bacterial growth. Thus, in this work, diameter controlled spherical Cucore-Agshell nanoparticles (Ag@CuNPs) with diameter ranging from 70 to 100 nm by one-step co-reduction approach were designed and synthesized. The Ag@CuNPs were homogenous, stable, and positively charged. The 70 nm Ag@CuNPs showed a consistent and regular Ag shielding. We observed the 100 nm Ag@CuNPs achieved symmetrical doped Ag clusters on the Cu core surface. We used Gram-positive and Gram-negative models strains to test the wide-spectrum antibacterial activity. The Ag@CuNPs showed detrimental microbial viability in a dose-dependent manner; however, 70 nm Ag@CuNPs were superior to those of 100 nm Ag@CuNPs. Initially, Ag@CuNPs attached and translocated the membrane surface resulting in bacterial eradication. Our analyses exhibited that antibacterial mechanism was not governed by the bacterial genre, nonetheless, by cell type, morphology, growing ability and the NPs uptake capability. The Ag@CuNPs were highly tolerated by human fibroblasts, mainly by the use of starch as glucosidic capper and stabilizer, suggesting optimal biocompatibility and activity. The Ag@CuNPs open up a novel platform to study the potential action of bimetallic nanoparticles and their molecular role for biomedical, clinical, hospital and industrial-chemical applications.

Simple impedimetric sensor for rapid lipase activity quantification.

A simple and rapid impedimetric sensor applicable for industrial lipase activity quantification was developed and characterized. It is based on the lipase catalyzed degradation by hydrolysis of a thin nanocomposite substrate sensitive layer deposited on a PCB stick electrode usable as disposable or regenerable. The sensitive layer degradation rate was evaluated by the impedance changes registration along time resulting from its thickness diminution applying a small amplitude AC voltage with a constant frequency. The AC current phase shift variations along the time caused by the impedance changes were registered as s sensor response. The sensor was characterized in terms of linear quantification range, LOD, precision and quantification time. The response time was found to be from 80 to 6 s for the linear concentration range from 0.99x10-2 to 1.68 U.S.P. U mL-1 with relative errors from 3.75% to 1.24% respectively and a LOD of 8x10-3 U.S.P. U mL-1. Finally, lipase spiked whey samples taken from milk industry were quantified and the results were validated by a titrimetric method revealing a good agreement (relative error less than 4.5%).

Rapid reagent-less on-line H2O2 quantification in alkaline semiconductor etching solution, Part 2: Nephelometry application.

A simple and rapid reagent less nephelometric method for on-line H2O2 quantification in semiconductors etching solutions was developed, optimized, characterized and validated. The intensity of the light scattered by the oxygen gas suspension resulted from H2O2 catalytic decomposition by immobilized MnO2 was registered as analytical response. The influences of the light wave length, the agitation rate, the temperature and the catalyst surface area on the response amplitude were studied and optimization was done. The achieved linear concentration range from 10 to 150mmolL-1 at 0.9835 calibration curve correlation coefficient, precision from 3.65% to 0.95% and response time from 35 to 20s respectively, at sensitivity of 8.01µAmmol-1 L and LOD of 2.9mmolL -1 completely satisfy the semiconductor industry requirements.

Immobilized microbial biosorbents for heavy metals removal.

Intensive industrial and urban growth has led to the release of increasing amounts of environmental pollutants. Contamination by metals, in particular, deserves special attention due to their toxicity and potential to bioaccumulate via the food chain. Conventional techniques for the removal of toxic metals, radionuclides and precious metals from wastewater all have a number of drawbacks, such as incomplete metal extraction, high cost and risk of generating hazardous by-products. Biosorption is a cost-effective and environment-friendly technology, an alternative to conventional wastewater treatment methods. Biosorption is a metabolically independent process, in which dead microbial biomass is capable of removal and concentrating metal ions from aqueous solutions. Free microbial biosorbents are of small size and low density, insufficient mechanical stability and low elasticity, which causes problems with metal ion desorption, separation of the sorbent from the medium and its regeneration. Hence, the possibilities for the implementation of continuous biosorbent processes for metal removal in flow-type reactor systems are reduced and the practical application of biosorption in industrial conditions is limited. By immobilizing microbial biomass on suitable carriers the disadvantages of free biosorbents are eliminated and more opportunities for practical use of biosorption become available. This review examines different immobilization techniques and carriers, certain basic features and possibilities of using immobilized microbial biosorbents for the removal and concentration of metals from aqueous solutions.

Rapid reagent-less on-line H2O2 quantification in alkaline semiconductor etching solution.

A simple, rapid, and reagent-less calorimetric method for H2O2 quantification, applicable automatically on-line was developed, analytically characterized and tested with real SC-1 alkaline etching solutions used in the semiconductor technology. Being based on H2O2 catalytic decomposition by immobilized solid catalyst, the proposed method possesses excellent specificity toward the H2O2 without any interference. The total quantification time was found to be less than 60s, RSD lies in the range from 4.7% to 1.8% for the linear concentration range from 2.8 10-2 to 2molL-1 respectively and the LOD was determined to be as low as 9.3 10-3molL-1.

In Vitro Assessment of Early Bacterial Activity on Micro/Nanostructured Ti6Al4V Surfaces.

It is imperative to understand and systematically compare the initial interactions between bacteria genre and surface properties. Thus, we fabricated a flat, anodized with 80 nm TiO2 nanotubes (NTs), and a rough Ti6Al4V surface. The materials were characterized using field-emission scanning electron microscopy (FE-SEM), energy dispersive X-ray spectroscopy (EDX) and atomic force microscopy (AFM). We cultured in vitro Staphylococcus epidermidis (S. epidermidis) and Pseudomonas aeruginosa (P. aeruginosa) to evaluate the bacterial-surface behavior by FE-SEM and viability calculation. In addition, the initial effects of human osteoblasts were tested on the materials. Gram-negative bacteria showed promoted adherence and viability over the flat and rough surface, while NTs displayed opposite activity with altered morphology. Gram-positive bacteria illustrated similar cellular architecture over the surfaces but with promoted surface adhesion bonds on the flat alloy. Rough surfaces supported S. epidermidis viability, whilst NTs exhibited lower vitality. NTs advocated promoted better osteoblast organization with enhanced vitality. Gram-positive bacteria suggested preferred adhesion capability over flat and carbon-rich surfaces. Gram-negative bacteria were strongly disturbed by NTs but largely stimulated by flat and rough materials. Our work proposed that the chemical profile of the material surface and the bacterial cell wall characteristics might play an important role in the bacteria-surface interactions.

Glucose Oxidase Biosensor Modeling and Predictors Optimization by Machine Learning Methods.

Biosensors are small analytical devices incorporating a biological recognition element and a physico-chemical transducer to convert a biological signal into an electrical reading. Nowadays, their technological appeal resides in their fast performance, high sensitivity and continuous measuring capabilities; however, a full understanding is still under research. This paper aims to contribute to this growing field of biotechnology, with a focus on Glucose-Oxidase Biosensor (GOB) modeling through statistical learning methods from a regression perspective. We model the amperometric response of a GOB with dependent variables under different conditions, such as temperature, benzoquinone, pH and glucose concentrations, by means of several machine learning algorithms. Since the sensitivity of a GOB response is strongly related to these dependent variables, their interactions should be optimized to maximize the output signal, for which a genetic algorithm and simulated annealing are used. We report a model that shows a good generalization error and is consistent with the optimization.

Detection of glycoalkaloids using disposable biosensors based on genetically modified enzymes.

In this work we present a rapid, selective, and highly sensitive detection of α-solanine and α-chaconine using cholinesterase-based sensors. The high sensitivity of the devices is brought by the use of a genetically modified acetylcholinesterase (AChE), combined with a one-step detection method based on the measurement of inhibition slope. The selectivity was obtained by using butyrylcholinesterase (BChE), an enzyme able to detect these two toxins with differential inhibition kinetics. The enzymes were immobilized via entrapment in PVA-AWP polymer directly on the working electrode surface. The analysis of the resulting inhibition slope was performed employing linear regression function included in Matlab. The high toxicity of α-chaconine compared to α-solanine due to a better affinity to the active site was proved. The inhibition of glycoalkaloids (GAs) mixture was performed over AChE enzyme wild-type AChE and BChE biosensors resulting in the detection of synergism effect. The developed method allows the detection of (GAs) at 50 ppb in potato matrix.

Comparison of the performances of four hydrophilic polymers as supports for lipase immobilisation.

Four hydrophilic polymers in the form of beads - chitosan, alginate, alginate/polyvinyl alcohol (PVA), and chitosan-coated alginate - were used as supports for lipase immobilisation. Hydrogel beads were characterised by bead-size-distribution estimation, surface morphology studies, and polymer interactions assessment. Matrix performances - loading efficiency, immobilisation yield, enzyme activity, and stability retention - were evaluated and compared. Although the loading efficiency of the chitosan-coated Ca-alginate beads (79.8%) was inferior to that of the Ca-alginate (87%) and of the Ca-alginate/PVA beads (81.3%), their enzyme immobilisation yield (63.96%) was the most important. Moreover, lipase encapsulated in chitosan-coated Ca-alginate beads demonstrated better pH, thermal, and storage (89% residual activity after 30 days) stabilities. Immobilised lipase activity also increased in the order: alginate/PVA > chitosan > alginate > alginate/chitosan, and displayed a maximum at pH 8 and at temperatures of 45 °C (chitosan and Ca-alginate/PVA beads) and 50 °C (Ca-alginate and chitosan-coated Ca-alginate beads). Thus, chitosan-coated Ca-alginate beads could be considered as a suitable support for lipase immobilisation.

LabVIEW 2010 Computer Vision Platform Based Virtual Instrument and Its Application for Pitting Corrosion Study.

A virtual instrumentation (VI) system called VI localized corrosion image analyzer (LCIA) based on LabVIEW 2010 was developed allowing rapid automatic and subjective error-free determination of the pits number on large sized corroded specimens. The VI LCIA controls synchronously the digital microscope image taking and its analysis, finally resulting in a map file containing the coordinates of the detected probable pits containing zones on the investigated specimen. The pits area, traverse length, and density are also determined by the VI using binary large objects (blobs) analysis. The resulting map file can be used further by a scanning vibrating electrode technique (SVET) system for rapid (one pass) "true/false" SVET check of the probable zones only passing through the pit's centers avoiding thus the entire specimen scan. A complete SVET scan over the already proved "true" zones could determine the corrosion rate in any of the zones.

Some clinical applications of the electrochemical biosensors.

Electrochemical biosensing, due to its sensitivity and specificity, combined with the low-cost and operation convenience of the equipment, is considered as a promising point-of-care approach in clinical analysis. This review presents the basic principles of operation, the current status, and the trends in the development and the clinical implementation of some selected electrochemical biosensors. These include: electrochemical glucose biosensors successfully applied in diabetes management, and electrochemical biosensors for cholinesterases and trypsin activities determination. The latter, although less common, demonstrate the potential of improving the existing clinical methods in the diagnostics and the treatment of neurotoxic, neurological, and pancreatic diseases.

In vitro Actinomyces israelii biofilm development on IUD copper surfaces.

BACKGROUND: Female pelvic actinomycosis may involve fallopian tubes, ovaries, uterus and bladder. This condition is often associated with the use of intrauterine contraceptive devices (IUDs), vaginal pessaries and/or tampons. The predominant causative agent of human actinomycosis is Actinomyces israelii, which has been found on copper IUDs retrieved from patients. STUDY DESIGN: In this work, a biofilm of A. israelii was developed in vitro on copper surfaces immersed in a simulated uterine fluid under anaerobic conditions. The biofilm was characterized using scanning electron microscopy (SEM), energy dispersive X-ray and atomic force microscopy. RESULTS: The capacity of A. israelii to develop a biofilm over copper surfaces in synthetic media was demonstrated. SEM micrographies illustrate the exopolysaccharides production and bacterial distribution. CONCLUSION: A. israelii was able to attach and grow in synthetic intrauterine media and to present on the copper surface is likely due to the production of biofilm.

Leptospirillum ferrooxidans based Fe2+ sensor.

A novel electrochemical biosensor integrating the strictly autotrophic bacterial strain Leptospirillum ferrooxidans as a recognition element and a Clark type oxygen probe as a transducer was designed, metrologically and analytically characterized and applied for the specific Fe(2+) determination. The bacterial Fe(2+) oxidation involves O(2) consumption, thus the quantification was performed registering the decrease of the oxygen reduction current. The limit of detection was found to be 2.4 micromol L(-1) and the sensitivity of the determinations-3.94 nAL micromol(-1). The response time of the biosensor is 18s for Fe(2+) concentrations of 10(-5) to 10(-4) mol L(-1). The biosensor was applied as well for the indirect determination of Fe(2+) oxidizing species such as Cr(2)O(7)(2-), reaching a sensitivity of 2.47 nAL micromol(-1). The transducer characteristics were evaluated and optimized to obtain short response time and high sensitivity. The analytical performances of the biosensor subject of the present work were found to be similar to that of the At. ferrooxidans based one developed by the authors earlier, avoiding however the sulfur compounds interference, because of the substrate specificity of the applied bacterial strain.

Electrochemical sensor based on Arthrobacter globiformis for cholinesterase activity determination.

The sensors applied recently for determination of cholinesterase activity are mostly enzymatic amperometric sensors, in spite of their disadvantages: short life-time at ambient temperature, instability of the response, interferences, as well as passivation of the electrode surface. In the present paper a new approach for determination of cholinesterase activity was proposed, overcoming the main drawbacks of the analysis performed with amperometric enzymatic sensors. Instead of the immobilization of enzymes on a conducting electrode surface, whole cells of Arthrobacter globiformis, containing choline oxidase were fixed on a Clark type oxygen probe. Current proportional to bacteria respiration is registered as a sensor response. The application of whole cells of bacteria as a sensing element permits to achieve high stability of the response and long life-time of the sensor at ambient temperature, due to the conservation of the enzyme in its natural micro-environment inside the immobilized cells. The proposed sensor keeps its functionality more than 7 weeks stored in deionized water at ambient temperature. For the first 2 weeks the amplitude of the response decreases with only 10% and at the end of the studied 7 weeks period the response was 50% of the initial. The other advantages of the proposed sensor are: the dissolved oxygen is used as a mediator which concentration can be reliably and interferences free measured by the aim of a Clark type oxygen probe applied as a transducer; reproducible bacterial membranes can be elaborated by filtration of resuspended bacterial culture after preliminary determination of its activity; application of membranes containing lyophilized bacteria capable to be conserved infinitely long time and activated just before their application; negligible cost compared with the sensors based on immobilized enzymes. The steady-state response of the proposed bacterial sensor to choline obtained in 200 s is linear in the investigated concentration range up to 2 x 10(-4) moldm(-3), with detection limit of 8 x 10(-8) moldm(-3) and sensitivity of 4 x 10(-1) microAcm(3)mol(-1), at pH 6, temperature of 25 degrees C and stirring rate of 300 rpm. Choline is formed as a result of the catalytic hydrolysis (depending on the cholinesterase activity) of the substrate acetylcholine. Linear calibration graph for cholinesterase activity determination was obtained in the range up to 11 mUcm(-3), with a slope of 1.97 x 10(-2) microAcm(3)mU(-1), at pH 6, temperature of 25 degrees C and stirring rate of 300 rpm. The tests with reconstituted lyophilized serum with known activity used as a control sample confirm the accuracy of the proposed method. The relative error of the determination was only 2.82%.

Bacterial sensors based on Acidithiobacillus ferrooxidans Part II. Cr(VI) determination.

The aerobic acidophilic bacterium Acidithiobacillus ferrooxidans oxidizes Fe(2+) and S(2)O(3)(2-) ions by consuming oxygen. An amperometric biosensor was designed including an oxygen probe as transducer and a recognition element immobilized by a suitable home-made membrane. This biosensor was used for the indirect amperometric determination of Cr(2)O(7)(2-) ions owing to methods based on a mediator (Fe(2+)) or titration. Using the mediator, the biosensor response versus Cr(2)O(7)(2-) was linear up to 0.4 mmol L(-1), with a response time of, respectively, 51 s (2 x 10(-5) mol L(-1) Cr(2)O(7)(2-)) and 61 s (6 x 10(-5) mol L(-1) Cr(2)O(7)(2-)). The method sensitivity was 816 microA L mol(-1). Response time and measurement sensitivity depended on membrane material and technique for biomass immobilization. For example, their values were 90 s-200 microA L mol(-1) when using a glass-felt membrane and 540 s-4.95 microA L mol(-1) with a carbon felt one to determine a concentration of 2 x 10(-5) mol L(-1) Cr(2)O(7)(2-). For the titration method, the biosensor is used to determine the equivalence point. The relative error of quantitative analysis was lower than 5%.

Bacterial sensors based on Acidithiobacillus ferrooxidans Part I. Fe2+ and S2O32- determination.

An amperometric bacterial sensor with current response to Fe(2+) and S(2)O(3)(2-) ions has been designed by immobilizing an acidophilic biomass of Acidithiobacillus ferrooxidans on a multi disk flat-front oxygen probe. The bacterial layer was located between the oxygen probe and a membrane of cellulose. A filtration technique was used to yield the bacterial membranes having reproducible activity. The decrease of O(2) flow across the bacterial layer is proportional to the concentration of the dosed species. The dynamic range appeared to be linear for the Fe(2+) ions up to 2.5 mmol L(-1) with a detection limit of 9 x 10(-7) mol L(-1) and a sensitivity of 0.25 A L mol(-1). The response of the biosensor is 84 s for a determination of 2 x 10(-4) mol L(-1) Fe(2+). Optimizing the Fe(2+) determination by A. ferrooxidans sensor was carried out owing to Design of Experiments (DOE) methodology and empirical modelling. The optimal response was thus obtained for a pH of 3.4, at 35 degrees C under 290 rpm solution stirring. S(2)O(3)(2-) concentration was determined at pH 4.7, so avoiding its decomposition. The concentration range was linear up to 0.6 mmol L(-1). Sensitivity was 0.20 A L mol(-1) with a response time of 207 s for a 2 x 10(-4) mol L(-1) S(2)O(3)(2-) concentration.

Acidithiobacillus ferrooxidans fixation on mercuric surfaces and its application in stripping voltammetry.

The adsorption (fixation) of bacteria Acidithiobacillus ferrooxidans on Hg and Cu metallic surfaces was qualitatively studied owing to two independent methods: frequency measurement using a quartz crystal microbalance and light absorption measuring at the Hg/bacterial culture interface. A method using a dropping mercury electrode (DME) allowed quantifying this bacterial fixation. Determining the superficial tension at Hg/bacterial culture interface led to determine bacteria adsorption on Hg through the Gibbs isotherm. A modified stripping voltammetry was proposed taking benefit of both bacterial adsorption on Hg surface and metal fixation capacity on biomass. Metal preconcentration on the biologically modified Hg electrode appeared to improve the measurement sensitivity of differential pulse anodic stripping voltammetry (DPASV). The height of the detected peaks was thus increased of 17.6% for copper, 48.4% for lead, and 132% for cadmium determinations compared to those obtained with an unmodified mercury electrode. Such augmentation depended on bulk bacteria concentration and bacteria preconcentration.