A new interference-free lysine biosensor using a non-conducting polymer film

An electrochemical biosensor for the determination of lysine to be used for rapid evaluation of food quality has been developed. Platinum electrodes have been coated by electropolymerisation with 1,2-diaminobenzene (1.2-DAB) using cyclic voltammetry. The reduction in the oxidation of interferents compared with the bare platinum electrode was 100% for ascorbic acid, 99% for acetaminophen and 99% for cysteine. The enzyme L-lysine-alpha-oxidase was then immobilised onto the polymer layer by passive adsorption and a calibration curve for lysine constructed. This gave a linear range of 1 x 10(-5) mol/l to 1 x 10(-3) mol/l and a limit of detection of 2 x 10(-7) mol/l.     

Detection of digitalis compounds using a surface plasmon resonance-based biosensor

A prospective national investigation comprising 633 extremely low birthweight (ELBW) infants born alive in the 2-y period 1990-1992 with a birthweight of < or = 1000 g and gestational age of > or = 23 completed weeks was conducted regarding neurosensory outcome and growth. Three-hundred and sixty-two (98%) surviving ELBW infants were assessed at a median age of 36 months, using a specially designed protocol. At follow-up, mean height, weight and head circumference in both boys and girls were significantly lower than the reference values. The incidence of cerebral palsy was 7% among all children and 14%, 10% and 3% in children born at 23-24, 25-26 and > or = 27 gestational weeks, respectively. At least one obvious handicap was present in 14%, 9% and 3% of these three groups of children, respectively. After adjustment for gestational age, a significantly increased risk of handicap was found in children with intraventricular haemorrhage grade > or = 3 and/or periventricular leucomalacia and in children with retinopathy of prematurity stage > or = 3. The results show that more than 90% of ELBW children born at > or = 25 completed gestational weeks were without neurosensory handicap at 36 months of corrected age. In infants born at 23-24 weeks of gestation, both survival and long-term outcome were less favourable.     

Quantifying serum antiplague antibody with a fiber-optic biosensor

Several studies of patients with chronic cervical pain and chronic whiplash syndrome report a high frequency of oculomotor function derangements pointing towards brainstem involvement and/or default sensory input from neck afferents. In light of these reports, it seems important to investigate other patient groups with similar upper cervical spine disorders. In this study, voluntary eye motor performance was evaluated in 11 rheumatoid patients (RA) with upper cervical dislocation and a clearly noticed joint affection of the cervical spine. The results were compared with 6 RA patients without cervical engagement and normal individuals. Nine of the 11 patients with atlanto-axial dislocation showed pronounced oculomotor disturbances of smooth pursuits compared with only minor changes in the control group. The pattern of oculomotor dysfunction in patients with rheumatoid cervical dislocation indicates brainstem involvement, which may be an early sign of brainstem affection/myelopathy.     

Screening of compounds interacting with HIV-1 proteinase using optical biosensor technology

A high-resolution optical biosensor assay for screening of low-molecular-weight compounds, using an immobilized protein target, has been developed. HIV-1 proteinase was immobilized on the sensor surface by direct amine coupling and a variety of inhibitors and noninteracting reference drugs were applied to the sensor surface in a continuous flow of buffer. The procedure did not require intrinsic reporter groups, substrates, inhibitors, or other ligands for detection. By using a reference protein, the signal could be corrected for the relatively large background signal caused by differences in dimethyl sulfoxide concentration between running and sample buffers. Substances binding with high affinity (Ki in nM range) required efficient regeneration of the sensor surface and washing of the injection system between sample cycles to get consistent results. Analysis was simplified by using report points, extracted during both association and dissociation phases, and a simple graphical display of data. The optimized assay could correctly distinguish HIV-1 inhibitors from other compounds in a randomized series, indicate differences in their interaction kinetics, and reveal artifacts due to nonspecific signals, incomplete regeneration, or carryover. The method is expected to be generally applicable to secondary screening of low-molecular-weight compound libraries with proteins as targets.     

Rapid monitoring of virus-like particles using an optical biosensor: a feasibility study

The mature major microneme protein of Sarcocystis muris cyst merozoites, which is known as a dimeric lectin with high affinity to galactose and some of its derivatives, was expressed in Escherichia colias a histidine-tagged fusion protein. The recombinant polypeptide, which was recognized by a monoclonal antibody directed against the native lectin, was purified from inclusion bodies after solubilization and refolding, using a combination of metal chelate and lactose affinity chromatography. The apparent molecular mass of the refolded polypeptide as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoreses was 16 kDa, whereas gel filtration chromatography clearly demonstrated that the recombinant protein, like its native counterpart, exists as a homodimer of two non-covalently associated subunits. Inhibition of haemagglutination suggests that the combining site of the recombinant lectin recognizes N-acetyl-galactosamine as the dominant sugar, thus confirming the correct folding of the monosaccharide combining site in the renatured lectin. To the best of our knowledge, this work represents the first reported detailed characterization of a recombinant lectin from apicomplexan parasites, and may contribute to a better understanding of the process of host cell recognition and invasion by these obligate intracellular protozoa.     

Hodgkin's disease: a tumor with disturbed immunological pathways

An experimental testing program was conducted to investigate the uplift capacity of various rafter to top plate connections. Rather than simply testing the fastener in an ideal test specimen configuration, attention is paid to more realistic test specimens that account for as-built conditions. Consideration is also given to repetitive systems of these connections such as would be found in roof systems. The results from this investigation provide basic data on failure modes and capacities. Comparisons are made between test values, calculated design values, and manufacturers' published values. In addition to metal straps, alternative connection methods such as adhesives are investigated, and consideration is given to both new construction and retrofit of existing structures. The results from this study are used to develop possible schedules for rafter to top plate connections considering different roof slopes, rafter lengths, wind speeds, exposures, and internal pressures.     

Determination of the step duration of gait initiation using a mechanical energy analysis

The analysis of gait initiation (the transient state between standing and walking) is an important diagnostic tool in the study of pathologic gait and the evaluation of prosthetic devices. Therefore it is important to know the step duration of gait initiation. However, there is little agreement in the literature regarding this step duration, since each author has based their conclusion on a different biomechanical parameter. In this study, gait initiation in seven normal subjects was studied using a mechanical energy analysis. The number of steps necessary to reach steady state was determined based on the fact that in steady-state gait, the net mechanical work of the body over one stride is zero (Winter et al. J. Biomechanics 9, 253-257, 1976). The variance of the work for a stride during steady-state walking was calculated for 100 steady-state trials from a separate database of normal subjects. The stride work was normalized to the subject's body weight (BW) and leg length (LL), and 95% confidence limits were defined from this data at -1.68%BW * LL < epsilon < 1.28%BW * LL. Total body energy during gait initiation was then computed for the seven test subjects. The energy analysis of gait initiation showed that steady state was attained by the end of three full steps. Therefore, a researcher studying gait initiation must allow his/her subject to take three full steps when recording data to ensure that the full event is included.     

Detection of protein A produced by Staphylococcus aureus with a fiber-optic-based biosensor

Staphylococcus aureus is a pathogen important in causing human infections and intoxication. A sensitive fiber-optic that produces evanescent waves was developed for the detection of protein A, a product secreted only by S. aureus. In the immunosensor, a 40-mV argon-ion laser that generated laser light at 488 nm was used together with plastic optical fiber and antibodies to protein A were physically adsorbed onto the fiber. The principle of the detection involved a sandwich immunoassay with fluorescein isothiocyanate conjugated with anti-(protein A) immunoglobulin G to produce signals of the antigen-antibody reaction. The detection limit was 1 ng of protein A per milliliter. The fiber-optic immunosensor could be used for rapid and specific detection of S. aureus in clinical specimens and foods.     

Identification of receptor ligands and receptor subtypes using antagonists in a capillary electrophoresis single-cell biosensor separation system

A capillary electrophoresis system with single-cell biosensors as a detector has been used to separate and identify ligands in complex biological samples. The power of this procedure was significantly increased by introducing antagonists that inhibited the cellular response from selected ligand-receptor interactions. The single-cell biosensor was based on the ligand-receptor binding and G-protein-mediated signal transduction pathways in PC12 and NG108-15 cell lines. Receptor activation was measured as increases in cytosolic free calcium ion concentration by using fluorescence microscopy with the intracellular calcium ion indicator fluo-3-acetoxymethyl ester. Specifically, a mixture of bradykinin (BK) and acetylcholine (ACh) was fractionated and the components were identified by inhibiting the cellular response with icatibant (HOE 140), a selective antagonist to the BK B2 receptor subtype (B2BK), and atropine, an antagonist to muscarinic ACh receptor subtypes. Structurally related forms of BK were also identified based on inhibiting B2BK receptors. Applications of this technique include identification of endogenous BK in a lysate of human hepatocellular carcinoma cells (Hep G2) and screening for bioactivity of BK degradation products in human blood plasma. The data demonstrate that the use of antagonists with a single-cell biosensor separation system aids identification of separated components and receptor subtypes.     

Production and manipulation of elevated temperatures in pig abdomen with a three-electrode capacitive heating device

The ability of a three-electrode capacitive heating device operating at 13.56 MHz to produce and effect spatial redistribution of hyperthermic temperatures is demonstrated through abdominal heating studies conducted with 80 kg female and 65 kg male pigs. Temperature-time profiles over the respective heating periods were obtained in anterior and posterior superficial tissue and in selected abdominal organs. At selected times during heating, spatial redistribution of elevated temperatures was effected through adjustment of power settings for one or more of the three electrodes, i.e. through manipulation of the specific absorption rate (SAR) in-vivo.     

Prediction of the creep behaviour of metallic materials under step loading using a simple mechanical model

The creep exposure of metallic parts usually occurs under time-variable loading conditions. For the prediction of creep behaviour under step loading a simple mechanical model is employed. The model is based on the time dependence of the material structure during creep. Therefore, the characteristic parameters of the model are considered to be functions of time and are correlated with the time dependence of the material structure parameters. For the definition of these functions a structure mechanical model has been employed. The mean internal material resistance has been considered as an appropriate structure parameter. The characteristic parameters of the model are defined in each step loading on the basis of the change of the internal stress derivative. For the verification of the model prediction the numerical results are compared with experimental results of the austenitic steel X 8 CrNiMoNb 16 16 for the orthogonal cyclic stress function.     

Analysis of lecithin in pharmaceutical products and diet integrators using a new biosensor operating directly in non aqueous solvent

One of the first examples of a bienzymatic organic phase enzyme electrode (OPEE) is described. It was obtained using two enzymes (phospholipase D and choline oxidase), both immobilised in kappa-Carrageenan gel and, as electrochemical transducer, an amperometric gas diffusion electrode for oxygen. The response of the biosensor was recorded and its sensitivity, linearity range, response time evaluated. Lastly drugs and diet products containing lecithin were analysed using the new biosensor device working in an organic mixture.     

Teaching in the ivory tower, with rarely a step outside.

Methods used in teaching psychology and in appraising course outcomes have been evaluated in a restricted manner. A broader base of evaluation criteria is recommended, including laboratory tasks, projects, field trips, and field experiences. Psychologists should not limit suggestions of needed research in this area to an ivory-tower ascension of more psychological theory, but should examine also what students, psychologists, and other folks are doing in the outside world. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Evaluation of lactate measurement in blood and plasma with biosensor technology: a comparison of methods

The introduction of biosensor technology for near bedside measurement of plasma lactate concentrations has been a promising step for critical care profiling. However, methodological drawbacks and relevant inaccuracy have been reported. With the advent of a new biosensor (Chiron Diagnostics) and a revised NOVA Biomedical device, accuracy was expected to be improved. The goal of the present investigation was to evaluate the accuracy of both methods. METHODS: Two devices (System 860, Chiron Diagnostics; StatProfile 9, NOVA Biomedical) were simultaneously analysed using 9 biosensors in both fresh frozen plasma and citrated whole blood. The results were compared with an established photometric method (Lactat PAP, Analyticon). Measurements were performed as duplicates (n = 1120) before and after the addition of 1 molar sodium lactate solution (2-24 mmol/L). For the estimation of between-day precision commercially available aqueous and serum-based quality controls were analysed daily over a period of 60 days. RESULTS: Reproducibility in blood was 2.6 +/- 2.8% (Chiron), 4.1 +/- 4.0% (NOVA) and 1.5 +/- 2.1% (Analyticon), in plasma respectively 2.1 +/- 2.4%, 2.1 +/- 2.9% and 1.0 +/- 1.1%. Mean inaccuracy in plasma presented to be -0.2 +/- 16.4% (plasma) and +7.2 +/- 13.1% (blood) for Chiron, +9.4 +/- 18.4% and +18.7 +/- 16.7% for NOVA, and -37.8 +/- 18.2% and -27.5 +/- 17.6% for Analyticon. Calculated between-day-precision (variation coefficients mean values) was 11.5 +/- 4.9% (Chiron) and 14.0 +/- 5.9% (NOVA). CONCLUSION: Although accuracy of lactate concentrations obtained with biosensor technology has improved (mean 0-18%), the variability of the results still poses a problem (mean 13-18%). Therefore, from the methodological point of view, interpretation of a single lactate value requires caution when applying to the critically ill, particularly with view to threshold values, and should be considered vis-à-vis other options.