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Biomedical subjects

A P Turner

Publications and source records attributed to A P Turner.

At least 19 recordsLinked to original sources

Immunosensor for 2,4-dichlorophenoxyacetic acid in aqueous/organic solvent soil extracts.

The development of a simple electrochemical immunoassay procedure for the field-based quantification of the herbicide 2,4-D in methanolic soil extracts is presented. The sensor utilizes a competitive immunoassay format incorporating an immobilized antigen complex at the surface of a disposable screen-printed working electrode element. The extent of glucose oxidase-labeled antibody binding to the antigen--electrode is determined amperometrically and is related to sample analyte concentration. The performance of the sensor is assessed in buffer, 30% methanol, and methanolic soil extracts. The device is capable of quantifying 2,4-D in all three matrixes at the low ppm level with coefficient of variation values of 6.2-33.6%. The causes of the variation observed in the sensor response in different soil matrixes are examined and improvements proposed. The sensor, tested in parallel with a commercial 2,4-D immunoassay test kit, yields comparable quantitative data and detection limits while exhibiting greater assay simplicity.

2,4-Dichlorophenoxyacetic Acid

The synthesis and screening of a combinatorial peptide library for affinity ligands for glycosylated haemoglobin.

This paper reports the synthesis and screening of a combinatorial peptide library for new affinity ligands for glycosylated haemoglobin (HbA1c), which is an important indicator of diabetes control. The new ligands are suitable for large-scale synthesis and overcome the disadvantages of antibodies (unstable and expensive to produce etc.), while remaining as efficient as antibodies in binding to the analyte. The library consisted of 262,144 hexapeptides synthesised using the one-bead-one-compound technique. The hexapeptides attached onto beads were screened with glycosylated haemoglobin HbA1c. The structures of the peptides exhibiting high affinity were characterised by Edman microsequencing. Computer modelling simulation of one of the lead sequences has shown that this class of ligand has a high affinity and specificity for glycosylated haemoglobin.

Animals

Immunomagnetic separation with mediated flow injection analysis amperometric detection of viable Escherichia coli O157.

The coupling of an immunological separation (using immunomagnetic beads) with amperometric flow injection analysis detection of viable bacteria is presented. Using a solution containing Escherichia coli O157, the electrochemical response with two different mediators [potassium hexacyanoferrate(III) and 2,6-dichlorophenolindophenol] was evaluated in the FIA system. Antibody-derivatized Dynabeads were used to selectively separate E. coli O157 from a matrix. The kinetics and the capacity parameters regarding the attachment of bacteria to the immunobeads were studied. The immunomagnetic separation was then used in conjunction with electrochemical detection to measure the concentration of viable bacteria. A calibration curve of colony-forming units (cfu) against electrochemical response was obtained. The detection limit for this rapid microbiological method was 10(5) cfu mL-1, and the complete assay was performed in 2 h. Some advantages over ELISA methods are the direct detection of viable cells (and not total bacterial load) and the need for only one antibody (not enzyme-labeled), thus making the assay faster (only one washing step is necessary) and less expensive.

2,6-Dichloroindophenol

Long-term outcomes of an arthritis self-management programme.

OBJECTIVE: A previous UK evaluation of the Arthritis Self-Management Programme (ASMP) demonstrated 4 month improvements in physical and psychological well-being including increased arthritis self-efficacy and increased use of self-management behaviours such as cognitive symptom management, and reductions in pain, fatigue and anxiety. The purpose of this study was to determine whether these effects were maintained at 12 month follow-up. METHODS: Twelve month data were collected via self-administered questionnaires mailed to participants who had previously responded prior to attending the ASMP and at 4 months follow-up. RESULTS: The sample (n = 112) comprised 82% women with a mean age of 59.6 (S.D. 12.4) yr and a mean disease duration of 14.9 (S.D. 11.1) yr. The majority of participants had a general practitioner-recorded diagnosis of either rheumatoid arthritis (46%) or osteoarthritis (44%). Many of the changes noted at 4 months were sustained at the 12 month follow-up. CONCLUSION: This first long-term evaluation of a community-based patient education intervention delivered in the UK suggests that after participation in the ASMP, persons with arthritis derive substantial and prolonged benefits in terms of perceived ability to manage arthritis, reduction in pain and improved psychological well-being.

Activities of Daily Living

Comparison of clinical and self-reported diagnoses for participants on a community-based arthritis self-management programme.

OBJECTIVE: With the advent of community-based arthritis education programmes, it is important to determine the accuracy of participants' self-reported diagnoses. The purpose of this study was to determine the level of agreement between general practitioner (GP)-recorded and self-reported diagnoses of participants attending an Arthritis Self-Management Programme (ASMP). METHODS: Participants enrolling on the ASMP were asked to (a) identify their type of arthritis via a self-administered postal questionnaire and (b) obtain a written confirmation of their diagnosis from their GP. The sample (n = 613) comprised mainly women (83%) with a mean age of 58.8 yr (S.D. 12.6) and a mean disease duration of 15.4 yr (S.D. 12.5). RESULTS: Participants' self-reported diagnoses were confirmed by GPs in 534 cases [87.1%, 95% confidence interval (CI): 84.4 89.8%]. Confirmed diagnoses were reported by 86.9% (95% CI: 83.1-90.7%) of those with osteoarthritis (OA) and 96.1% (95% CI: 93.6 98.6%) of those with rheumatoid arthritis (RA). The concordance rate for all other types of arthritis combined was lower at 60.5% (95% CI: 49.5-71.5%). There were no significant differences with respect to age, gender, education, physical functioning, duration of disease and number of GP visits between those who correctly identified their type of arthritis and those who did not. CONCLUSIONS: This study suggests that the majority of RA and OA participants attending an arthritis education programme can correctly identify their specific type of arthritis.

Adult

Microbial detection.

There is a widespread need for commercial instrumentation for the rapid and inexpensive detection of microbial contamination of food, industrial waste water and clinical samples. A large number of detection methods have been developed utilizing the optical, electrochemical, biochemical and physical properties of microorganisms. The need for a device which can produce a rapid, accurate, sensitive, real-time analysis for clinical, industrial and environmental applications has led to considerable progress being achieved in recent years in the development of biosensors for microbial detection. This intense research has resulted in the commercialization of several instruments. Techniques used for the quantification of microorganisms are reviewed under the general categories of non-bioelectrochemical and bioelectrochemical methods.

Bacteriological Techniques

A new approach for creating double-stranded DNA biosensors.

The principle of 'sandwich'-type biosensors based on liquid-crystalline dispersions formed from [DNA-polycation] complexes is outlined. These biosensors will find application in the determination of a range of compounds and physical factors that affect the ability of a given polycationic molecule to maintain intermolecular crosslinks between neighbouring DNA molecules. In the case of liquid-crystalline dispersions formed from [DNA-protamine] complexes, the lowest concentration of hydrolytic enzyme (trypsin) detectable was approximately 10-(14) M.

Animals

On-line monitoring of glucose, glutamate and glutamine during mammalian cell cultivations.

Amperometric biosensors (based on rhodinised carbon electrodes) for glucose, glutamine and glutamate were constructed. The sensors were incorporated into a three cell parallel FIA system and used to monitor the three analytes on-line during two mammalian cell perfusion cultures. All measurements were made simultaneously from undiluted media sample. Use of the FIA system enabled easy and rapid exchange of the sensors, during cultivation. The inclusion of a calibration step, regularly for all sensors, helped to maintain the accuracy of all measurements. Comparison with off-line measurements indicated that all three biosensors operated successfully, providing accurate information.

Biosensing Techniques

Amperometric detection of histamine at a quinoprotein dehydrogenase enzyme electrode.

Methylamine dehydrogenase, a tryptophan tryptophyl quinone (TTQ) containing quinoprotein, catalyzes the oxidation of a variety of primary aliphatic monomaines and diamines to their respective aldehydes and ammonia. This paper reports the construction and characterization of an enzyme electrode capable of detecting histamine and methylamine at +200 mV versus a saturated calomel reference electrode. The methylamine dehydrogenase isolated from Paracoccus denitrificans was used in conjunction with the insoluble mediator tetracyanoquinodimethane (TCNQ) to construct enzyme electrodes which will potentially provide simple rapid analysis of histamine without the need for the extensive sample pretreatments currently required in HPLC and GLC analysis. The linear response of this amperometric sensor, between 0 and 200 microM, correlates well with elevated histamine levels predominant in patients with chronic myelogenous leukaemia, whilst the observed limit of detection, 4.8 microM, compares favourably with the lower limits of detection reported for a potentiometric histamine sensitive enzyme electrode.

Biosensing Techniques

Recent advances in amperometric glucose biosensors for in vivo monitoring.

Electrochemical biosensors for glucose, based on the specific glucose oxidizing enzyme glucose oxidase, have generated considerable interest. Several commercial devices based on this principle have been developed and are widely used for in vitro monitoring of glucose e.g. in hospitals, doctors surgeries and for home monitoring by patients themselves. A significant advance in the application of biosensor technology would be the development of portable, implantable sensors which could continuously indicate the blood glucose concentration, enabling swift corrective action to be taken by the patient. This review highlights recent developments in amperometric glucose biosensors for in vivo monitoring and also considers the remaining barrier which need to be overcome to enable successful introduction of an implantable sensor.

Biosensing Techniques

Electrochemical sensors for continuous monitoring during surgery and intensive care.

The current state of development of electrochemical sensors and biosensors for continuous use during surgery and intensive care is briefly reviewed with an emphasis on recent developments. The clinical usefulness of invasive and non-invasive sensors is discussed. Recent advances in the design of electrochemical sensors and experience with ex vivo and in vivo applications are described. The importance of developing appropriate fabrication technology is emphasised in order to meet the demand for reliable and reproducible analytical devices.

Animals

Biosensors.

Advances in biosensor technology over the past year have included developments in metalized electrodes, mediated electrochemistry, direct electron transfer, impedance measurement, optical immunosensors, optodes, biomimicry, piezoelectric biosensors, enzyme thermistors, in vivo biosensors, surface characterization, organic-phase biosensors and tissue-based biosensors. Increasingly, molecular biology and engineering are being used in sensor design.

Biomedical Engineering

The 3a protein from cucumber mosaic virus increases the gating capacity of plasmodesmata in transgenic tobacco plants.

The 3a protein, encoded by RNA 3 of cucumber mosaic virus (CMV), is the putative movement protein of viral progeny in infected plants. An analysis of transgenic tobacco plants constitutively expressing the CMV 3a protein showed that the protein is accumulated in leaves at every stage of development. In fully expanded leaves the protein is immunodetectable mostly in a cell-wall-enriched fraction. Dye-coupling experiments using fluorescent-dextran probes were performed on fully expanded leaves to study the modifying effect of CMV 3a protein on the gating capacity of plasmodesmata. Movement of fluorescein-isothiocyanate-labelled dextran with a mean molecular mass of 10,000 Da, and an approximate Stokes' radius of 2.3 nm, was detected between cells of the 3a protein transgenic plants, but not in the control plants. These results are consistent with the idea that the CMV 3a protein is involved in the modification of plasmodesmata and, therefore, in the cell-to-cell spread of the virus.

Blotting, Western

Effect of the alfalfa mosaic virus movement protein expressed in transgenic plants on the permeability of plasmodesmata.

Symplastic transport of different sized fluorescent probes has been assessed in leaf epidermal cells of transgenic Nicotiana plants expressing the movement protein (MP) of alfalfa mosaic virus (AMV). In both N. tabacum and N. benthamiana, the size exclusion limit (SEL) of plasmodesmata increased from M(r) 1000, which represents the commonly accepted limit, to over 4.4K. However, in control plants, movement of a 3K probe was seen in 11 to 22% of the injections, indicating that plasmodesmata may on occasion allow the passage of molecules larger than was previously thought. The increase of SEL due to the presence of the AMV MP, although significant, remains insufficient to permit the passage of viral particles and the possibility of other mechanisms involved in viral cell-to-cell spread is discussed.

Alfalfa mosaic virus

Involvement of a neutral glycolipid in differential cell adhesion in the Xenopus blastula.

Many different molecular species mediate cell adhesion during embryonic development. These can have either protein or carbohydrate functional groups, which can act in either a homophilic or a heterophilic manner, and often in concert. We report here that a monoclonal antibody, M4B, raised against Xenopus blastomere membranes, inhibits the calcium-dependent adhesion of dissociated blastomeres. M4B maintains its inhibitory effect on adhesion when converted into univalent fragments, and specifically affects calcium-dependent adhesion. The antigen is regulated in both space and time during early development. It is found on cell surfaces throughout the egg to blastula stages, but is more concentrated on cells in the animal and marginal zones of the blastula. It is dramatically downregulated during gastrulation, and becomes largely restricted to gut epithelium by the larval stages. We show also that M4B function is spatially differentiated at the blastula stage, since it inhibits the aggregation of dissociated animal cells to a greater extent than vegetal cells. This membrane antigen may therefore play a role in the differential adhesion observed between different regions of the blastula, and which we presume to underlie the segregation of the primary germ layers during gastrulation. M4B recognizes a complex of plasma membrane glycolipids. Periodate treatment destroys the ability of these glycolipids to react with the antibody, indicating that the epitope resides in the carbohydrate moiety of the glycolipids. Chemical characterization shows that it is a neutral glycolipid, and that the major component is of the glycoglycerolipid, rather than the more common glycosphingolipid class. Blocking experiments with oligosaccharides of defined structure, and antibody crossreactivity show that the M4B antibody does not recognize several known embryonic carbohydrate antigens. These results demonstrate that M4B antibody recognizes a novel group of developmentally regulated glycolipids which function in calcium-dependent cell--cell adhesion in the Xenopus blastula.

Animals