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At least 379 records · Page 21Linked to original sources

Electrokinetic control of fluid in plastified laser-printed poly(ethylene terephthalate)-toner microchips.

The application of plastified laser-printed poly(ethylene terephthalate)(PET)-toner microchips to capillary electrophoresis was investigated. Electroosmotic flow was observed in the direction of the cathode for the buffer system studied (phosphate, pH 3-10). Average electroosmotic mobilities of 1.71 x 10(-4) to 4.35 x 10(-4) cm(2) V(-1) s(-1) were observed from pH 3 to 10. This variation suggests that silica fillers in the toner and on the surface of the polymer dominate the zeta potential of the material, which is also confirmed by XPS measurements. Dopamine and catechol were used as model analytes for microchip electrophoresis in combination with electrochemical detection. Results show that these two analytes can be efficiently separated and detected electrochemically with the plastified laser-printed PET-toner microchips.

Journal Article↗

Flow screen-printed amperometric detection of p-nitrophenol in alkaline phosphatase-based assays.

p-Nitrophenyl phosphate is one of the most widely used substrates for alkaline phosphatase in ELISAs because its yellow, water-soluble product, p-nitrophenol, absorbs strongly at 405 nm. p-Nitrophenol is also electroactive; an oxidative peak at 0.97 V (vs. an Ag pseudoreference electrode) is obtained when a bare screen-printed carbon electrode is used. When an amperometric detector was coupled to a flow-injection analysis system the detection limit achieved for p-nitrophenol was 2x10(-8) mol L(-1), almost two orders of magnitude lower than that obtained by measuring the absorbance of the compound. By use of this electrochemical detection method, measurement of 7x10(-14) mol L(-1) alkaline phosphatase was achieved after incubation for 20 min. The feasibility of coupling immunoassay to screen-printed carbon electrode amperometric detection has been demonstrated by performing an ELISA for detection of pneumolysin, a toxin produced by Streptococcus pneumoniae, which causes respiratory infections. The method is simple, reproducible, and much more sensitive than traditional spectrophotometry.

Alkaline Phosphatase↗

Microfabrication of screen-printed nanoliter vials with embedded surface-modified electrodes.

A self-contained ion-selective sensing system within a nanoliter-volume vial has been developed by integrating screen printing, laser ablation, and molecular imprinting techniques. Screen printing and laser ablation are used in tandem to fabricate nanoliter-volume vials with carbon and Ag/AgCl ring electrodes embedded in the sidewalls. Using multisweep cyclic voltammetry, the surface of the carbon electrode can be modified with a polypyrrole film. By polymerizing pyrrole in the presence of nitrate, pores complementary to the nitrate anion in size, shape, and charge distribution are formed in the resulting film. Electrochemical cells modified with this nitrate-imprinted polypyrrole film show a near-Nernstian response to nitrate, and excellent reproducibility. The integration of molecular recognition and electrochemical response in the nanoliter vials is demonstrated by the detection of as little as 0.36 ng nitrate in nanoliter-volume samples. The integration of tailored molecular recognition within nanoliter vials via established fabrication and imprinting protocols should result in a number of nanosensor devices with applications in BioMEMS and micro total analysis systems.

Electrochemistry↗

NADH screen-printed electrodes modified with zirconium phosphate, Meldola blue, and Reinecke salt. Application to the detection of glycerol by FIA.

Bulk screen-printed electrodes (bSPEs) modified with zirconium phosphate (ZrP) and Meldola blue (MB) and by electrochemical deposition of a Reineckate film (bMBZrPRs-SPEs) have been constructed and used as NADH sensors. Cyclic voltammetric investigation of these bulk electrochemically modified screen-printed electrodes revealed stable catalytic activity in oxidation of the reduced form of the coenzyme nicotinamide adenine dinucleotide (NADH). Flow-injection analysis (FIA) coupled with amperometric detection confirmed the improved stability of the bMBZrPRs-SPEs (10(-4) mol L(-1) NADH, %RSD = 4.2, n = 90, pH 7.0). Other conditions, for example applied working potential (+50 mV relative to Ag|AgCl), flow rate (0.30 mL min(-1)) and pH-dependence (range 4.0-10.0) were evaluated and optimized. A glycerol biosensor, prepared by immobilizing glycerol dehydrogenase (GDH) on the working electrode area of a bMBZrPRs-SPE, was also assembled. The biosensor was most stable at pH 8.5 (%RSD = 5.6, n = 70, 0.25 mmol L(-1) glycerol). The detection and quantification limits were 2.8 x 10(-6) and 9.4 x 10(-6) mol L(-1), respectively, and the linear working range was between 1.0 x 10(-5) and 1.0 x 10(-4) mol L(-1). To assess the effect of interferences, and recovery by the probe we analyzed samples taken during fermentation of chemically defined grape juice medium and compared the results with those obtained by HPLC.

Beverages↗

Amperometric sensor for L-ascorbic acid determination based on MnO2 bulk modified screen printed electrode.

A simple biosensor constructed by bulk-modification of carbon ink with manganese dioxide as a mediator was investigated for its ability to serve as amperometric detector for L-ascorbic acid in hydrodynamic mode. The sensor could be operated at pH 5.0 (0.05 M phosphate buffer) and exhibited excellent reproducibility and stability. Optimization of measurement parameters such as applied working potential and pH value were studied in detail. The screen printed electrode exhibited a linear amperometric increase with the concentration of L-ascorbic acid from 50 mg L(-1) to 250 mg L(-1) and gave a (LOD = 3sigma) detection limit of 0.2 mg L(-1) (1.172 micromol L(-1)). The manganese dioxide modified screen printed electrode shows long term stability.

Ascorbic Acid↗

A screen-printed biosensor using pyruvate oxidase for rapid determination of phosphate in synthetic wastewater.

A screen-printed phosphate biosensor based on immobilized pyruvate oxidase (PyOD, E.C. 1.2.3.3) has been developed for monitoring phosphate concentrations in a sequencing batch reactor (SBR) system. The enzyme was immobilized by a nafion matrix and covered a poly(carbamoyl) sulfonate (PCS) hydrogel on a screen-printed electrode. PyOD consumes phosphate in the presence of pyruvate and oxygen and generates hydrogen peroxide (H2O2), carbon dioxide and acetylphosphate. The electroactive H2O2, monitored at +420 mV vs Ag/AgCl, is generated in proportion to the concentration of phosphate. The sensor has a fast response time (2 s) and a short recovery period (2 min). The time required for one measurement using this phosphate biosensor was 4 min, which was faster than the time required using a commercial phosphate testing kit (10 min). The sensor has a linear range from 7.5 microM to 625 microM phosphate with a detection limit of 3.6 microM. There was good agreement (R2=0.9848) between the commercial phosphate testing kit and the phosphate sensor in measurements of synthetic wastewater in a SBR system. This sensor maintained a high working stability (>85%) after 12 h of operation and involved a simple operation procedure. It therefore serves as a useful tool for rapid and accurate phosphate measurements in the SBR system and probably for process control.

Bioreactors↗

[Images of the elderly in printed advertisements in a time change perspective].

This content analytical study investigates the changes of image of the elderly in printed advertisements. A total of 364 advertisements published in 1999 and 2000 involving elderly models were analyzed. A positive image of older age is generally transported in advertisements addressing all consumers, whereas physical and cognitive deficits occurring with age are expressed in some verbal elements of advertisements addressing primarily the elderly themselves. However, such deficits are not visually depicted, since health, full energy and activity provide positive orientation. Compared with advertisements in the 1970s and 1980s, the elderly are shown as more attractive, active and socially appreciated today. In spite of their improved image, the elderly, especially older women, are still not represented in printed advertisement in proportion to their number in the population.

Activities of Daily Living↗

Hydroxyapatite scaffolds for bone tissue engineering made by 3D printing.

Nowadays, there is a significant need for synthetic bone replacement materials used in bone tissue engineering (BTE). Rapid prototyping and especially 3D printing is a suitable technique to create custom implants based on medical data sets. 3D printing allows to fabricate scaffolds based on Hydroxyapatite with complex internal structures and high resolution. To determine the in vitro behaviour of cells cultivated on the scaffolds, we designed a special test-part. MC3T3-E1 cells were seeded on the scaffolds and cultivated under static and dynamic setups. Histological evaluation was carried out to characterise the cell ingrowth. In summary, the dynamic cultivation method lead to a stronger population compared to the static cultivation method. The cells proliferated deep into the structure forming close contact to Hydroxyapatite granules.

3T3 Cells↗

Tissue print micropeel: a new technique for mapping tumor invasion in prostate cancer.

Because of widely adopted screening programs for early detection of prostate cancers, many patients who undergo radical prostatectomy have tumors that are not grossly evident, and the extent and distribution of the cancer in the gland can only be determined by a microscopic examination of the surgical specimen. Historically, one of the most important predictors of the quality of cancer control following surgical resection of a solid tumor is the absence of cancer at the surgical margins. Although the clinical significance of cancer at the margins of a radical prostatectomy specimen has been a source of controversy in recent years, surgical pathology assessment of radical prostatectomy margins remains an important part of prostate cancer clinical care. However, a comprehensive histopathologic review of every radical prostatectomy specimen is beyond the resources of most hospitals. Tissue print micropeel technologies, combined with appropriate markers, provide a new strategy that combines a relatively simple technique for sampling specimen margins with a method for obtaining molecular information about the cancer that can add to the macroscopic and microscopic anatomical findings. This new tissue printing approach for incorporating molecular markers into the assessment of radical prostatectomy margins is reviewed in this article.

Biomarkers, Tumor↗

A high-affinity protein stain for western blots, tissue prints, and electrophoretic gels.

A method for protein staining using copper phthalocyanine 3,4',4'',4'''-tetrasulfonic acid tetrasodium salt is described. The procedure is applicable to protein blots and tissue prints, as well as to polyacrylamide and agarose gels. It is also simple, involving only application of the stain and rinsing. For protein blots and tissue prints the staining is rapid, taking less than 1 min to completion, and more sensitive than any previously described dye-based nonspecific protein staining system. The staining is easily reversible, requiring only a change in pH to remove the dye.

Blotting, Western↗

Interpreting palmar sweat prints.

The plastic impression method to assess the number of active palmar sweat glands (PSI) was used to study changes of sweat gland activity during several phases of dental treatment. Subjects were 64 female and 38 male patients at a dental surgery. At the beginning they scaled five typical dental situations in terms of perceived threat: waiting-room before treatment: sitting down in the dental chair; dentist enters and starts treatment; after treatment in the dental chair; and back in waiting-room after treatment. Four prints were taken in the waiting-room before treatment and one in each other situation described above. The average of prints 3 and 4 was used for phase comparisons. Palmar sweat index values were generally in accordance with the threat of the situations. In particular there was a rise from waiting-room to sitting down in the dental chair and a further increase at the dentist's entry, whereafter values dropped considerably. A significant decrease from the first two waiting-room values to the following two suggests that the PSI reacts sensitively to the novelty of the assessment procedure itself and that the very first recordings should hence not be taken into evaluation.

Adolescent↗

Improved imaging of rat hindfoot prints for walking track analysis.

Walking track analysis is an investigative technique that allows a researcher to assess objectively the functional capacity of the limb in the rat sciatic nerve model. This study has shown that paint and paper is a better method for imaging the rat foot plantar surface than other methods described in the literature. The block printing paint has the distinct advantage of keeping radial diffusion error to a minimum. Paint more correctly images the important anatomical landmarks of the rat plantar surface. There is better traction (less slippage) rendering the prints easier to interpret. The authors conclude that the paint and paper is a superior method to the other methods currently available.

Animals↗

Non-isotopic tissue-printing hybridization: a new technique to study long-distance plant virus movement.

A non-isotopic tissue-print hybridization technique was developed to study long-distance plant virus movement. By using digoxigenin-labeled RNA probes the distribution pattern of the viral RNA was observed in leaf, stem and petiole tissues. In leaf tissue viral RNA was confined preferentially to symptoms and veins, and in stem and petiole sections, the hybridization signal was observed in vascular tissue. Both chemiluminescent and colorigenic detection methods were used. The colorigenic method, though less sensitive, is advantageous in that it gives some anatomical information on the signal distribution. This non-isotopic tissue-print hybridization technique can provide considerable information about the spatial and temporal virus expression with regard to its symptoms.

Digoxigenin↗

Measurement of soluble L-lactate in dairy products using screen-printed sensors in batch mode.

Amperometric sensors based on lactate oxidase and platinized carbon were constructed entirely by screen-printing. They were used to estimate the lactate concentration in diluted samples of yoghurt and buttermilk. Estimates made in untreated (apart from dilution) solutions were subject to bias, relative to values obtained from spectrophotometric assays. Removal of cations improved the accuracy of the estimates. Calcium may have made a small contribution to the bias but the nature of the inhibition is otherwise unknown. With untreated samples, determinations of any accuracy require sufficient dilution to avoid such bias, although the smaller signals resulting from dilution place more demands on calibration and manufacturing standards. The relative standard deviations of estimates, based on variation among sensors, of the most dilute samples were generally too high for practical use. The errors of estimates at lower dilutions approached those of screen-printed sensors made under industrial conditions.

Biosensing Techniques↗

Electrochemical detection of E. coli 16S rDNA sequence using air-plasma-activated fullerene-impregnated screen printed electrodes.

A new DNA modified electrode for the electrochemical detection of 16S rDNA extracted from Escherichia coli (JCM1649) is proposed. The electrodes were fabricated by screen printing a fullerene-impregnated carbon ink onto a poly(methylmethacrylate) substrate and immobilizing a probe DNA on the surface after activating the electrode with air plasma. The results indicated a dramatic improvement in the surface coverage of the immobilized probe DNA, and of the reduction peak of the redox indicator (Co(phen)(3)(3+)) due to the incorporation of fullerene. By immobilizing the probe onto the fullerene-impregnated screen-printed electrodes, the PCR product of the 16S rDNA extracted from E. coli was directly detected without any pretreatment. A well defined signal difference was observed between the perfectly matching oligonucleotide and the mismatching one, and it was possible to detect the target at the modified electrode. This method enabled us to clearly detect the two base mismatches in the ca. 1500-bases long 16S rDNA sequence.

Air↗

Oligonucleotide-modified screen-printed gold electrodes for enzyme-amplified sensing of nucleic acids.

An electrochemical genosensor for the detection of specific sequences of DNA has been developed using disposable screen-printed gold electrodes. Screen-printed gold electrodes were firstly modified with a mixed monolayer of a 25-mer thiol-tethered DNA probe and a spacer thiol, 6-mercapto-1-hexanol (MCH). The DNA probe sequence was internal to the sequence of the 35S promoter, which sequence is inserted in the genome of GMOs regulating the transgene expression. An enzyme-amplified detection scheme, based on the coupling of a streptavidin-alkaline phosphatase conjugate and biotinylated target sequences was then applied. The enzyme catalysed the hydrolysis of the electroinactive alpha-naphthyl phosphate to alpha-naphthol; this product is electroactive and has been detected by means of differential pulse voltammetry. The assay was, firstly, characterised using synthetic oligonucleotides. Relevant parameters, such as the probe concentration and the immobilisation time, the use of the MCH and different enzymatic conjugates, were investigated and optimised. The genosensor response was found to be linearly related to the target concentration between 0 and 25 nmol/L; the detection limit was 0.25 nmol/L. The analytical procedure was then applied for the detection of the 35S promoter sequence, which was amplified from the pBI121 plasmid by polymerase chain reaction (PCR). Hybridisation conditions (i.e., hybridisation buffer and hybridisation time) were further optimised. The selectivity of the assay was confirmed using biotinylated non-complementary amplicons and PCR blanks. The results showed that the genosensor enabled sensitive (detection limit: 1 nmol/L) and specific detection of GMO-related sequences, thus providing a useful tool for the screening analysis of bioengineered food samples.

Alkaline Phosphatase↗

An electrochemical immunosensor for aflatoxin M1 determination in milk using screen-printed electrodes.

The production and assembling of disposable electrochemical AFM1 immunosensors, which can combine the high selectivity of immunoanalysis with the ease of the electrochemical probes, has been carried out. Firstly immunoassay parameters such as amounts of antibody and labelled antigen, buffer and pH, length of time and temperature of each steps (precoating, coating, binding and competition steps) were evaluated and optimised in order to set up a spectrophotometric enzyme-linked immunosorbent assay (ELISA) procedure. This assay exhibited a working range between 30 and 160 ppt in a direct competitive format. Then electrochemical immunosensors were fabricated by immobilising the antibodies directly on the surface of screen-printed electrodes (SPEs), and allowing the competition to occur between free AFM1 and that conjugated with peroxidase (HRP) enzyme. The electrochemical technique chosen was the chronoamperometry, performed at -100 mV. Furthermore, studies of interference and matrix effects have been performed to evaluate the suitability of the developed immunosensors for the analysis of aflatoxin M1 directly in milk. Results have shown that using screen-printed electrodes aflatoxin M1 can be measured with a detection limit of 25 ppt and with a working range between 30 and 160 ppt. A comparison between the spectrophotometric and electrochemical procedure showed that a better detection limit and shorter analysis time could be achieved using electrochemical detection.

Aflatoxin M1↗

Electrochemical immunosensor array using a 96-well screen-printed microplate for aflatoxin B1 detection.

A novel analytical immunosensor array, based on a microtiter plate coupled to a multichannel electrochemical detection (MED) system using the intermittent pulse amperometry (IPA) technique, is proposed for the detection of aflatoxin B1 (AFB1). In the present work, the electrochemical behaviour and electroanalytical performance of the thick-film carbon sensors (also designated as screen-printed electrodes) incorporated in the multichannel electrochemical plate were first evaluated. Then the 96-well screen-printed microplate was modified in accord with a competitive indirect enzyme-linked immunoassay (ELISA) format for aflatoxin B1 detection. The measurements were performed using both spectrophotometric and electrochemical procedures and the results of the calibration curves, detection limit (LOD), sensitivity and reproducibility of the respective assay systems were evaluated. The immunoassay was then applied for analysis of corn samples spiked with AFB1 before and after the extraction treatment, in order to study the extraction efficiency and the matrix effect, respectively. These studies have shown that using this system, AFB1 can be measured at a level of 30 pg/mL and with a working range between 0.05 and 2 ng/mL. Good recoveries (103+/-8%) were obtained, demonstrating the suitability of the proposed assay for accurate determination of the AFB1 concentration in corn samples. The specificity of the assay was assessed by studying the cross-reactivity of PAb relative to AFB1. The results indicated that the PAb could readily distinguish AFB1 from other aflatoxins, with the exception for AFG1.

Aflatoxin B1↗