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Biomedical subjects

Neil Harris

Publications and source records attributed to Neil Harris.

15 recordsLinked to original sources

A microarray-based detection system for genetically modified (GM) food ingredients.

A multiplex DNA microarray chip was developed for simultaneous identification of nine genetically modified organisms (GMOs), five plant species and three GMO screening elements, i.e. the 35S promoter, the nos terminator and the nptII gene. The chips also include several controls, such as that for the possible presence of CaMV. The on-chip detection was performed directly with PCR amplified products. Particular emphasis was placed on the reduction of the number of PCR reactions required and on the number of primers present per amplification tube. The targets were biotin labelled and the arrays were detected using a colorimetric methodology. Specificity was provided by specific capture probes designed for each GMO and for the common screening elements. The sensitivity of the assay was tested by experiments carried out in five different laboratories. The limit of detection was lower than 0.3% GMO for all tests and in general around 0.1% for most GMOs. The chip detection system complies with the requirements of current EU regulations and other countries where thresholds are established for the labelling of GMO.

Agrobacterium tumefaciens↗

How expert are the experts? An exploration of the concept of 'expert' within Delphi panel techniques.

The use of the term 'expert' occurs widely in healthcare research, in the context of national guidelines and consensus methods for the development of clinical protocols. Within consensus methods of research, especially Delphi panel techniques, the use of 'experts' is fundamental to reliability. Yet literature fails to debate the practicalities of defining 'experts' for use within Delphi panel research. This paper, by John Baker and colleagues, draws on methodological literature and discusses the concepts and elements of 'experts'. It concludes with recommendations for researchers to ensure experts for future Delphi research.

Delphi Technique↗

Standardisation of data from real-time quantitative PCR methods - evaluation of outliers and comparison of calibration curves.

BACKGROUND: As real-time quantitative PCR (RT-QPCR) is increasingly being relied upon for the enforcement of legislation and regulations dependent upon the trace detection of DNA, focus has increased on the quality issues related to the technique. Recent work has focused on the identification of factors that contribute towards significant measurement uncertainty in the real-time quantitative PCR technique, through investigation of the experimental design and operating procedure. However, measurement uncertainty contributions made during the data analysis procedure have not been studied in detail. This paper presents two additional approaches for standardising data analysis through the novel application of statistical methods to RT-QPCR, in order to minimise potential uncertainty in results. RESULTS: Experimental data was generated in order to develop the two aspects of data handling and analysis that can contribute towards measurement uncertainty in results. This paper describes preliminary aspects in standardising data through the application of statistical techniques to the area of RT-QPCR. The first aspect concerns the statistical identification and subsequent handling of outlying values arising from RT-QPCR, and discusses the implementation of ISO guidelines in relation to acceptance or rejection of outlying values. The second aspect relates to the development of an objective statistical test for the comparison of calibration curves. CONCLUSION: The preliminary statistical tests for outlying values and comparisons between calibration curves can be applied using basic functions found in standard spreadsheet software. These two aspects emphasise that the comparability of results arising from RT-QPCR needs further refinement and development at the data-handling phase. The implementation of standardised approaches to data analysis should further help minimise variation due to subjective judgements. The aspects described in this paper will help contribute towards the development of a set of best practice guidelines regarding standardising handling and interpretation of data arising from RT-QPCR experiments.

Biotechnology↗

Healthy living interventions and schizophrenia: a systematic review.

AIM: This paper reports a systematic review of the published and grey literature which has investigated the efficacy of healthy living interventions for adults with a diagnosis of schizophrenia or schizo-affective disorder. BACKGROUND: Adults with a diagnosis of schizophrenia or schizo-affective disorder have reduced life expectancy when compared with members of the general population, with approximately 59% of excess mortality resulting from natural causes. METHODS: The review was conducted following guidelines provided by the United Kingdom National Health Service Centre for Reviews and Dissemination, and using the Medline, PsycINFO, CINAHL, Embase, Cochrane Library, National Research Register, and System for Info on Grey Literature databases. RESULTS: Sixteen studies were identified, examining four types of healthy living interventions: smoking cessation (n = 7), weight management (n = 5), exercise (n = 3) and nutritional education (n = 1). The smoking cessation, weight management and exercise studies showed positive outcomes in the main. The quality of the studies, however, was generally poor. Only two had control groups, most recruited small self-selected samples, six did not standardize for diagnosis, external validity was generally poor and no studies followed participants for longer than 6 months. The best quality evidence was produced by the smoking cessation and weight management studies, which were more methodologically robust and demonstrated promising outcomes. CONCLUSIONS: Further research is needed to assist the development of effective interventions to help this client group to adopt and maintain healthier lifestyles. Research and practice development in this area may be an important role for nurses in both hospital and community settings.

Health Promotion↗

The ADVIA 2120 hematology system: flow cytometry-based analysis of blood and body fluids in the routine hematology laboratory.

The ADVIA 2120 Hematology System was recently released by Bayer HealthCare, Diagnostics Division, as a bench-top analyzer designed for medium- to large-volume laboratories. This flow cytometry-based system uses light scatter, differential white blood cell (WBC) lysis, and myeloperoxidase and oxazine 750 staining to provide a complete blood cell count, a WBC differential, and a reticulocyte count. A cyanide-free method is used to measure hemoglobin colorimetrically. The system is automation ready; in addition to its capability for analyzing peripheral blood specimens, the analyzer is also equipped to analyze cerebrospinal fluid samples. In this article we explain the underlying technology of the ADVIA 2120, provide linearity ranges, method-specific reference ranges, and stability data, and describe novel parameters and applications that are unique to the methodology used by this instrument. Finally, we discuss research applications and future directions, such as the use of this hematology analyzer in the determination of fetal lung maturity.

Blood Cell Count↗

Performance evaluation of the ADVIA 2120 hematology analyzer: an international multicenter clinical trial.

Automated cell counters are widely used in modern clinical laboratories to provide reliable, fast, and cost-effective complete blood counts (CBCs), white blood cell differentials, and reticulocyte measurements. In addition, some advanced instruments provide novel parameters, such as the hemoglobin content of reticulocytes or the percentage of hypochromic cells, and are capable of analysis of a variety of body fluids. Bayer recently introduced the ADVIA 2120 system as an automation-ready cell counter for mid- to high-volume testing in the clinical laboratory. This instrument, which builds on the established technology of the ADVIA 120 system, operates with a cyanide-free method for hemoglobin measurement, has a new user interface, and can routinely analyze biological fluid samples in addition to blood. We used 749 samples from 6 worldwide trial sites to evaluate the clinical performance of this new device. Accuracy of the ADVIA 2120 system versus its predecessor model, the ADVIA 120 system, was excellent for all CBC and white cell differential parameters and reticulocyte counts (all correlation coefficients except for basophils >0.9). Correlation of the white cell differential with the standard manual method and within-run precision of the ADVIA 2120 system also was very good. Use of the novel cyanide-free method for hemoglobin measurement had no clinically significant impact on hemoglobin results, even in patients with hemoglobinopathies. We concluded that the ADVIA 2120 system has clinically equivalent performance to the ADVIA 120 system.

Automation↗

The synthesis and SAR of 2-arylsulfanyl-phenyl piperazinyl acetic acids as glyT-1 inhibitors.

Elevation of glycine levels and activation of the NMDA receptor by inhibition of the glycine transporter 1 (GlyT-1) is a potential strategy for the treatment of schizophrenia. A novel series of GlyT-1 inhibitors have been identified containing the 2-arylsulfanyl-phenylpiperazine motif. The most prominent member of this series, (R)-4-[5-chloro-2-(4-methoxy-phenylsulfanyl)-phenyl]-2-methyl-piperazin-1-yl-acetic acid (31) is a potent glycine transporter-1 inhibitor (IC(50)=150 nM), which elevated glycine levels in rat ventral hippocampus as measured by microdialysis in vivo at doses of 1.2-4.6 mg/kg s.c.

Acetates↗

Analysis and interpretation of data from real-time PCR trace detection methods using quantitation of GM soya as a model system.

Recent years have seen an increased interest in DNA trace detection methods involved in many areas of bioanalytical research, such as quantitation of genetically modified (GM) ingredients in food products. There is little in the way of standardisation of data handling from these methods, and the data generated needs to be analysed appropriately if the results are to be interpreted correctly. This paper describes particular aspects of real-time PCR trace detection methods in order to increase the understanding of data generated using this bioanalytical technique. Using the specific example of GM soya detection and quantitation, it focuses on the production of calibration curves based on the mean and individual data values, the interpretation of correlation coefficients, regression techniques, and discusses suitable data analysis arising from simple and more complex experimental designs following transformation. By using the approaches outlined in this paper, more accurate analysis of data from real-time PCR and GM trace detection methods could be achieved.

Calibration↗

Development and evaluation of patient-centered software for a weight-management clinic.

OBJECTIVE: To describe a weight-management clinic software system and to report on its preliminary evaluation. RESEARCH METHODS AND PROCEDURES: The software system standardizes the collection of relevant patient information from an initial medical assessment, weekly clinic visits, and laboratory testing protocol of a medically supervised proprietary meal-replacement program in a university-based referral clinic. It then generates monthly patient feedback reports with graphs of clinical and laboratory parameters to support a patient-centered approach to weight management. After patients and clinic physicians review the data to ensure accuracy, the database is used for subsequent patient feedback reports, reports to referring physicians, quality assurance, and research. Clinic physicians and referring physicians were asked to rate their acceptance of the system. In addition, in a retrospective analysis of data generated by the system, outcomes for patients who received system-generated feedback (n = 620) were compared with those who participated in the program before the introduction of feedback (n = 130). RESULTS: Clinic and referring physicians reported that they had high overall satisfaction with the software and that the system saved them time, and the latter group reported that it decreased laboratory use. Regarding patients, the feedback group had lower dropout rates in the latter half of the program, better rates of attendance, completion of laboratory tests, and weight loss after 8 weeks. DISCUSSION: The software seems to facilitate the effectiveness of the treatment protocol for obesity and generates a high-quality database for patient care, clinic administration, quality assurance, and research purposes.

Ambulatory Care↗

Parental expectations of care and charges in a tertiary care pediatric ED.

Our objective was to assess parental expectations of diagnostic testing, time, and charges in a pediatric emergency department (PED) using a prospective survey-based study. Parents on arrival estimated the amount of testing, time, and charges expected to occur. A total of 266 of 294 (90.5%) of the questionnaires were completed and returned. Parents correctly predicted the need for radiographic evaluation in 185 of 258 (71.7%), laboratory evaluation in 179 of 257 (69.6%), and medication in 144 of 259 (55.6%). The average length of stay was 2 hours and 36 minutes and was correctly estimated within preset ranges by 86 of 253 (34.0%) and underestimated by 112 of 253 (44.3%). The mean total charge was $964 per visit and was correctly estimated within preset ranges by 91 of 260 (35.0%) and underestimated by 139 of 260 (53.3%). Despite being relatively accurate about the need for medications and radiographic and laboratory studies, parents are still more likely than not to underestimate the total charges and time associated with a PED visit.

Adult↗

Type 1 diabetes islet autoantibody markers.

The diagnosis of type 1 diabetes versus other forms of diabetes such as type 2 diabetes is paramount to guiding proper therapy. Several islet autoantibodies have been identified that serve to diagnose immune-mediated, type 1a diabetes in clinically ambiguous cases. These autoantibodies also serve to predict type 1 diabetes in nondiabetic individuals. The most useful islet autoantibodies include islet cell cytoplasmic autoantibodies, insulin autoantibodies, glutamic acid decarboxylase autoantibodies, and insulinoma-associated-2 autoantibodies. Once type 1 diabetes can be safely and reliably prevented, large-scale islet autoantibody screening programs of the general pediatric population may be warranted. It is controversial whether islet autoantibodies influence the course of type 1 diabetes following diagnosis.

Animals↗

Detection of genetically modified crops and their derivatives: critical steps in sample preparation and extraction.

The detection of genetically modified crops in foodstuff relies on detection of transgenic DNA or protein material in the sample matrix. Purified DNA or proteins are used as analytical material for polymerase chain reaction technologies and immunodiagnostics. Successful sample preparation is critical to the validity of subsequent analysis. For routine analysis, a good sample preparation technique should be simple, safe, and inexpensive while reproducibly generating DNA/protein of sufficient quality and yield. The suitability of isolated DNA or protein as an analyte for a detection or characterization technique depends on amount or concentration, purity, and integrity, each of which may be influenced by sample matrix and the extraction technique, and, in turn, may impact the validity of analytical techniques. The key sample preparation steps of homogenization, pretreatment, extraction, and purification are discussed as well as typical analytical methods. Consideration is given to application of these steps for particular sample matrixes to maximize yield, reduce inhibition effects, and minimize contamination. The choice of the most appropriate and valid methods for sample preparation from particular foods is discussed with respect to DNA analysis. Attention is also given to ease of use, cost, and generic applicability of the procedures.

Crops, Agricultural↗

Real-time detection of genetically modified soya using Lightcycler and ABI 7700 platforms with TaqMan, Scorpion, and SYBR Green I chemistries.

A comparative cross platform evaluation of real-time polymerase chain reaction detection of DNA sequences present in Roundup Ready soya was undertaken using the ABI 7700 and Roche Lightcycler detection systems in combination with 3 different detection chemistries: TaqMan, Scorpion primers, and SYBR Green I fluorescent dye. Various copy numbers of a plasmid containing the soya lectin sequence were used to determine the sensitivity and reproducibility of the different technology combinations and to examine both inter and intra machine variability. To examine the relative accuracy of each technology, the genetically modified soya content of baked products containing known amounts of Roundup Ready soya was determined by detection of lectin and the EPSPS transgene. It was determined that the combination of TaqMan detection chemistry and the ABI 7700 platform represented the best method for quantitative detection of genetically modified organisms in terms of both precision and accuracy.

Analysis of Variance↗

Detection of genetic modification in cured tobacco leaf: proficiency testing using polymerase chain reaction-based methods.

The Cooperation Centre for Scientific Research Relative to Tobacco (CORESTA; Paris, France) "Task Force Genetically Modified Tobacco-Detection Methods" investigated the performance of qualitative and quantitative methods based on the polymerase chain reaction (PCR) for the detection and quantitation of genetically modified (GM) tobacco. In the 4 successful rounds of proficiency testing, the cauliflower mosaic virus 35S RNA promoter (CaMV 35S) and the Agrobacterium tumefaciens nopaline synthase terminator (NOS) were selected as target sequences. Blind-coded reference materials containing from 0.1 to 5.0% and from 0.15 to 4% GM tobacco were used in 2 rounds of qualitative and quantitative PCR, respectively. Eighteen laboratories from 10 countries participated in this study. Considering all methods and 2 rounds, the different laboratories were able to detect GM tobacco at the 0.1% level in 46 out of 58 tests in qualitative assays. The results of the proficiency test indicate that both end point screening and real-time quantitative methods are suitable for the detection of genetically modified organisms in tobacco leaf samples having a GM content of 0.1% or higher. The CORESTA proficiency study represents a first step towards the interlaboratory evaluation of accuracy and precision of PCR-based GM tobacco detection, which may lead to the harmonization of analytical procedures and to the enhancement of comparability of testing results produced by different laboratories.

Agrobacterium tumefaciens↗