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At least 1,279 records · Page 71Linked to original sources
PubMed1994

A chromatographic procedure for fully automated isolation of DNA from human whole blood.

Isolation of DNA from peripheral blood constitutes a fundamental step in the molecular diagnosis of genetic disorders, analysis of mutations of oncogenes and detection of nucleic-acid sequences of pathogenic organisms in clinical samples. We investigated whether dextran sedimentation of blood followed by the chromatographic isolation of DNA could be applied to the Biomek-1000 automated laboratory workstation in order to develop a fully automated method of DNA isolation from whole blood. Under our experimental conditions, the Biomek-1000 could perform fully automated DNA isolation from seven human whole blood samples in less than 1.5 h. Our data indicate that the DNA isolated from human blood in this way is suitable for polymerase chain reaction.

Autoanalysis↗
PubMed1987

An automated method for autoradiographic analysis of cultured Schwann cells.

A semi-automated analysis system based on video image analysis was developed to count labelled and unlabelled nuclei of Schwann cells which had been exposed to tritiated thymidine followed by processing for autoradiography. A Model 3000 Image Analysis system (Image Technology Corporation, Deer Park, NY) was used to acquire and process the images and provide quantitative measurements based on the distinctive size and shape of the Schwann cell nucleus. The maximum and minimum dimensions for the labelled and unlabelled nuclei were determined. These stored dimensional parameters were then compared with the dimensions of a given field of cell nuclei by the image analysis system. The counts from various fields were collected until a total of 1000 labelled and unlabelled nuclei had been analyzed. A labelling index (LI = ratio of labelled cells to total cells X 100) was then calculated and printed by the system. LIs of autoradiographs determined by automated analysis correlated well with those determined by visual cell counting. The principle of the image analysis program as described here is applicable to other systems for the measurement of LIs of a particular cell type in a mixed population. This automated process eliminates both the subjectivity and fatigue of visual counting and facilitates the rapid measurement of the LI of large numbers of autoradiographs with precision.

Animals↗
PubMed1984

Application of automated REM and slow wave sleep analysis: I. Normal and depressed subjects.

Computerized analysis of rapid eye movement (REM) and delta electroencephalographic (EEG) sleep patterns in normal and depressed subjects offers opportunities to examine sleep more precisely than previously possible. In the present study, automated REM analyses demonstrated good reliability with traditional manual procedures in both normal and depressed subjects. However, automated delta analyses correlated well with traditional scoring in normal subjects, but not in depressed patients. These findings suggest the use of automated delta techniques similar to those employed in this report or spectral analytic techniques in the following types of studies: specificity of delta sleep in various psychiatric syndromes, changes in delta sleep produced by the administration of psychotropic agents, relationships between delta sleep and sleep-related neuro-endocrine patterns, and, finally, relationships between delta sleep patterns and other biological rhythms such as activity and temperature.

Adult↗
PubMed1996

Automated RNA probe assay for the identification of Listeria monocytogenes.

Recent epidemics of human listeriosis have shown the importance of Listeria monocytogenes as a food-borne risk. We have developed an automated identification assay for L. monocytogenes using the specificity of the 16S rDNA probe described by Wang et al. (1991). Bacterial cultures were lysed quickly by a chemical step releasing the target nucleic acids. Extracts were loaded into the VIDAS automate (bioMérieux) which performed a non-radio-active hybridisation reaction for 2 h. This assay recognised specifically 68 out of 69 L. monocytogenes isolates from clinical and food origins, including serovars 4b and 1/2, without cross-hybridising to other Listeria species (103 strains) or other bacterial species (15 strains). The sensitivity of the assay was 10(7) bacteria. This automated technology is faster than conventional biochemical identification.

Base Sequence↗
PubMed1988

Information for decision makers at hospital laboratory level: an example of a graphical method of representing costs and effects for a replacement automated technology in a haematology laboratory.

The costs and effects on patient management following the introduction of a fully automated blood count analyser have been evaluated for a hospital haematology laboratory in England, during 1985. In the case of the relatively high throughput observed in the laboratory studied, the automated technology was judged to be cheaper if considering laboratory-centred costs alone. But this was no longer necessarily the case once wider costs, including extra medical workload generated for hospital and community physicians, were included. Effects on patient management were measured for all types of patient, including hospital in-patients, out-patients and general practice patients. A graphical method allowing comparison of the costs and effects of the replacement automated system, termed a Technology MAP (Management Awareness Profile) is presented in the paper. MAPs provide a flexible information display which can be tailored to an individual laboratory's situation producing an overall picture of expected costs and effects on patient management following the introduction of a new technology--a full value-in-use assessment. The authors argue that there is a need for a central source of such value-in-use assessments, especially for new medium-priced diagnostic technologies which are likely to become widely spread in Britain.

Blood Cell Count↗
PubMed1996

Evaluation of an automated real-time spectral analysis technique.

An adaptive real-time Doppler peak-frequency tracing algorithm was evaluated in vitro and compared to manual peak-frequency traces. A computer-controlled pump was used to generate physiological flow waveforms in a vasculature-mimicking phantom. Spectral waveforms were obtained on an ATL HDI along with real-time estimates of diagnostic parameters, including maximum systolic, minimum diastolic, time-averaged peak frequencies and pulsatility and resistance indices. The effect of the signal-to-noise ratio on the measured parameters was investigated. The imprecision in the measured parameters was found to depend somewhat on the waveform shape; e.g., the imprecision in PI was 4.1% for a normal renal waveform and 8.5% for a waveform having reverse diastolic flow. The peak frequency envelopes of the same waveform data were traced manually by nine operators, and the resulting diagnostic parameters were compared to ones obtained from automated peak-frequency traces of the same waveform data. The agreement between parameters measured by the automated routine and those measured manually was found to depend somewhat on the waveform shape; e.g., the bias in the PI was 1.3% for a renal waveform lacking diastolic flow, and 12% for a waveform with reverse diastolic flow. The between-observer variations in the manual measurements ranged from 0.8% up to 9.4%. The overall variations associated with the automated traces were found to be smaller than or equal to those of the manual traces.

Algorithms↗
PubMed1992

Determination of urinary normetanephrine, metanephrine and 3-methoxytyramine by high-performance liquid chromatography with electrochemical detection: comparison between automated column-switching and manual dual-column sample purification methods.

This report describes a high-performance liquid chromatographic method with electrochemical detection for the simultaneous quantitation of urinary metanephrine, normetanephrine and 3-methoxytyramine. This method, which involves manual dual-column purification steps for the routine determination of urinary metanephrines, is compared with the previously used spectrophotometric Pisano method and an on-line sample preparation procedure, where the automated sequential trace enrichment (ASTED) apparatus is used for the column-switching procedure. In order to automate the metanephrine assay, the enrichment technique was evaluated against the reference chromatographic method. Bio-Rad urine controls gave coefficients of variation of less than 9% at all levels for the reference method. Values of less than 19% were found in the reference range with the enrichment method, and the recovery of 3-methoxytyramine was also too poor to be measured in normal concentrations. The linearity of both methods is sufficient to determine pathological levels of these biogenic amines. Future developments should be focused on decreasing the variation of between-day assays in an on-line, automated procedure.

Adult↗
PubMed1993

Automated determination of free phenytoin in human plasma with on-line equilibrium dialysis and column-switching high-performance liquid chromatography.

Free phenytoin in human plasma was automatically determined by on-line equilibrium dialysis using the automated sequential trace enrichment of dialysate (ASTED) sample preparation system and HPLC. The dialysis cell was a modification of the cell supplied with the ASTED. Total phenytoin was analysed with the same analytical set-up and plasma protein binding was determined. Free phenytoin was determined in plasma from epileptic patients and the results were compared to those obtained by ultrafiltration. Automated determination of free and total phenytoin in plasma by the ASTED-HPLC combination was shown to be an accurate and reproducible method and the results in free phenytoin analyses were in agreement with those found with ultrafiltration. The sample throughput with the automated on-line combination of dialysis and column-switching HPLC was 75 samples in 24 h when the sample was dialysed at 37 degrees C.

Autoanalysis↗
PubMedDate not supplied

Determination of verapamil and norverapamil in human plasma by liquid chromatography: comparison between a liquid-liquid extraction procedure and an automated liquid-solid extraction method for sample preparation.

A conventional liquid-liquid extraction (LLE) procedure with high-performance liquid chromatography (HPLC) has been developed for the determination of verapamil and its main metabolite, norverapamil, in plasma. After addition of the internal standard, plasma samples were basified with phosphate buffer (pH 9.0) and extracted with a mixture of cyclohexane-dichloromethane. After centrifugation, the organic layer was separated and the analytes were extracted back into a 0.1 N sulphuric acid solution containing 2-aminoheptane. An aliquot of this aqueous phase was then injected directly onto the HPLC column. This LLE procedure has been compared with an automated liquid-solid extraction (LSE) method that has been developed in parallel. Good linearity was obtained using both extraction methods. The absolute recoveries for the two analytes were ca 95% with the automated LSE procedure and slightly lower (ca 84%) for the LLE method. The automated method gives better results with respect to detectability and precision, but the LLE procedure is simpler to develop, requires much less expensive equipment, and remains a useful alternative when the number of samples to be analysed is limited.

Buffers↗
PubMed1994

Automated analytical systems for drug development studies. I--A system for the determination of drug stability.

An automated system consisting of a pH-stat, microdialysis sampling and a liquid chromatograph was assembled to measure the rate of rapid chemical reactions. 2',3',5'-Triacetyl-6-azauridine was used as a model compound to validate the performance of the automated system. Buffer catalysis was minimized by using a non-catalytic concentration of borate buffer along with a pH-stat to maintain the pH during the kinetic run. The microdialysis sampling technique permitted sample quenching and buffering of the solutions to a pH compatible with the LC column materials. The combination of microdialysis sampling and rapid LC analysis allowed reactions with a half-life of approximately 1 min to be sampled every 30 s. The rates of hydrolysis of the drug, measured at different conditions of temperature (37-70 degrees C) and pH (9.0-10.5) using the automated system, compared well with the previously determined values.

Autoanalysis↗
PubMed1996

Fully automated assay for the determination of sumatriptan in human serum using solid-phase extraction and high-performance liquid chromatography with electrochemical detection.

A method is described for a fully automated, sensitive, accurate and precise assay for the determination of sumatriptan in human serum. The assay consists of solid-phase extraction followed by reversed-phase HPLC with electrochemical detection. The extraction procedure has been fully automated on a Zymate XP robot linked on-line to the HPLC system. The assay is linear over the analytical range 1-30 ng ml-1 and selective for sumatriptan with respect to endogenous plasma components and GR49336, the major circulating metabolite. The intra-assay data demonstrate a maximum bias and precision across the calibration range of 10% and 6.6% respectively. The inter-assay data demonstrate a maximum bias and precision across the calibration range of 6.7% and 8.8%, respectively. The extraction efficiency of the assay is approximately 90% and is constant across the calibration range. The assay was used for the determination of sumatriptan in serum clinical samples and was shown to be robust in sustained use over several months. The use of a Zymate XP robot allowed complete automation of the assay, which resulted in high-quality, high-throughput analyses.

Autoanalysis↗
PubMed1996

Comparative evaluation of an automated ribotyping system versus pulsed-field gel electrophoresis for epidemiological typing of clinical isolates of Escherichia coli and Pseudomonas aeruginosa from patients with recurrent gram-negative bacteremia.

Ribotyping and macrorestriction analysis of chromosomal DNA using pulsed-field gel electrophoresis (PFGE) are among the more useful molecular epidemiologic typing methods. Because these techniques are labor intensive, automation of one or more steps may allow clinical laboratories to apply molecular typing methods. We compared the recently developed automated ribotyping system, the RiboPrinter Microbial Characterization System (DuPont), with PFGE as a means of typing clinical isolates of E. coli and P. aeruginosa. A total of 22 E. coli and 24 P. aeruginosa were typed by both PFGE and the RiboPrinter. When compared with PFGE typing of E. coli and P. aeruginosa, the RiboPrinter was less sensitive in identifying different strains, particularly among the isolates of P. aeruginosa. The RiboPrinter was completely automated and allowed up to 32 isolates to be typed within an 8-hour period. The pattern of results obtained in this study suggests that a heirarchical approach to molecular typing using the RiboPrinter Microbial Characterization System plus PFGE might be feasible. The RiboPrinter Microbial Characterization System promises to be a very useful addition to the expanding molecular typing armamentarium.

Bacteremia↗
PubMed1992

Performance of the automated complete Selvester QRS scoring system in normal subjects and patients with single and multiple myocardial infarctions.

The automated version of the complete Selvester QRS scoring system for estimation of myocardial infarct size was evaluated in 1,344 normal subjects, 706 patients with a single myocardial infarction (366 with inferior infarction, 277 with anterior infarction and 63 with posterolateral infarction) and 131 patients with combined inferior and anterior infarction. The presence and location were determined by angiographic and ventriculographic criteria. The performance of the overall 32-point system, each of the 19 criteria and the 13 criteria sets and each of the 35 criteria within the 13 sets was examined. The mean point scores were 1.7 for normal subjects, 3.7 for posterolateral infarction, 4.1 for inferior infarction, 6.3 for anterior infarction and 6.9 for multiple infarcts. A score greater than 4 yielded a sensitivity of 67% for anterior infarction, 41% for inferior infarction, 32% for posterolateral infarction and 72% for multiple infarcts. However, 7 of 32 criteria failed to achieve 95% specificity and 10 of 35 criteria in criteria sets had a sensitivity that was even lower than their false positive rate. The automated Selvester QRS scoring system currently has limitations that are attributable to development of the original system, which used manual scoring techniques and established criteria limits from middle-aged men. Future automated analysis should use gender- and age-dependent criteria limits.

Computer Simulation↗
PubMed1993

An evaluation of automated defibrillation and manual defibrillation by emergency medical technicians in a rural setting.

We show that automated external defibrillation training of emergency medical technicians (EMTs) is less time consuming than manual defibrillation training, and hypothesize that both improve survival from sudden cardiac death. Data on 91 cardiac arrests over 27 months among five basic life support services was collected before EMT-defibrillation (EMT-D) training. Subsequently, seven BLS services were trained in EMT-D using either manual difibrillation or automated external defibrillation technology, and 55 sudden cardiac death patients were entered after training. Manual defibrillation required 11 more hours per student in initial training. Survival to hospital discharge improved from two of 91 patients (2.2%) in the series before EMT-D training to nine of 55 patients (16.4%) after EMT-D training (P = .001). Improved survival was correlated with shorter prehospital defibrillation times, 8.84 minutes, when EMTs performed defibrillation versus 16.3 minutes before training when EMTs awaited advanced life support defibrillation (P < .001). To enhance equipment familiarity we allowed EMTs to apply three-lead electrode monitors to all medical/cardiac patients during transport (surveillance). There were six emergency medical service-witnessed "surveillance" arrests and three arrests survived to hospital discharge (50% survival). This group represented 33% of all survivors in the series. We recommend automated external defibrillation training for EMTs. Improved survival in sudden cardiac death cases in well-run emergency medical service systems should result from EMT-D training. Finally, we recommend that routine "surveillance" of high-risk patients during transport by defibrillation-capable EMTs be considered in EMT-D programs, rather than limiting EMT-D only to units capable of rapid "man-down" response.

Aged↗
PubMed1988

Automated synthesis system with computer control for the production of [1-11C]fatty acids.

To accommodate the increasing need for [1-11C]fatty acids for use in routine medical studies and the large amounts of radioactivity required, a more advanced automated synthesis system for the preparation of [1-11C]fatty acids has been constructed. The synthesis of [1-11C]fatty acids has been completely automated using a new separation unit (isobaric Sepaltor) for the extraction procedure, which had caused some difficulties with automation. The yield of [1-11C]palmitic acid was 20-30 mCi, and that of 3-methyl [1-11C]heptadecanoic acid was 2-7 mCi. The time required for the synthesis was less than 30 min from the start of the 11CO2 trapping. This system has been used for more than 20 preparations of [1-11C]palmitic acid and more than 20 preparations of 3-methyl [1-11C]heptadecanoic acid.

Carbon Radioisotopes↗
PubMed1988

Automated production of several positron-emitting radiopharmaceuticals using a single laboratory robot.

A Zymate Laboratory Automation System (Zymark Corp.), previously set up for the automated synthesis of 16 alpha-[18F]fluoroestradiol-17 beta [Brodack et al. (1986a) J. Nucl. Med. 27, 714] has been modified for the production of several short-lived radiopharmaceuticals in a single hot cell. All manipulations and apparatus normally used in the syntheses of carbon-11 and fluorine-18-labeled radiopharmaceuticals have been incorporated into the robot system. This achievement permits facile modifications of existing procedures used by the robot in addition to the incorporation of new routines in a minimal amount of time. Currently, the Zymate robot is programmed for the routine production of 16 alpha-[18F]fluoroestradiol-17 beta (7-11% EOS in 80 min), N-(3-[18F]fluoropropyl)spiperone (15-18% EOS in 70 min), and [1-11C]butanol (11-15% EOS in 25 min). A fourth compound, 2-deoxy-2-[18F]fluoro-D-glucose, is also synthesized by the robot. The yields and synthesis times of the robot-produced compounds are comparable to those obtained during manual syntheses. This method of automation represents a flexible and versatile alternative for the routine production of radiopharmaceuticals used in PET studies.

Fluorine Radioisotopes↗
PubMed1992

A chronic disease score from automated pharmacy data.

Using population-based automated pharmacy data, patterns of use of selected prescription medications during a 1 year time period identified by a consensus judgement process were used to construct a measure of chronic disease status (Chronic Disease Score). This score was evaluated in terms of its stability over time and its association with other health status measures. In a pilot test sample of high utilizers of ambulatory health care well known to their physicians (n = 219), Chronic Disease Score (CDS) was correlated with physician ratings of physical disease severity (r = 0.57). In a second random sample of patients (n = 722), its correlation with physician-rated disease severity was 0.46. In a total population analysis (n = 122,911), it was found to predict hospitalization and mortality in the following year after controlling for age, gender and health care visits. In a population sample (n = 790), CDS showed high year to year stability (r = 0.74). Based on health survey data, CDS showed a moderate association with self rated health status and self reported disability. Unlike self-rated health status and health care utilization, CDS was not associated with depression or anxiety. We conclude that scoring automated pharmacy data can provide a stable measure of chronic disease status that, after controlling for health care utilization, is associated with physician-rated disease severity, patient-rated health status, and predicts subsequent mortality and hospitalization rates. Specific methods of scoring automated pharmacy data to measure global chronic disease status may require adaptation to local prescribing practices. Scoring might be improved by empirical estimation of weighting factors to optimize prediction of mortality and other health status measures.

Adolescent↗
PubMedDate not supplied

Automated texture-based segmentation of ultrasound images of the prostate.

Segmenting two-dimensional images of the prostate into prostate and nonprostate regions is required when forming a three-dimensional image of the prostate from a set of parallel two-dimensional images. The texture-based segmentation method presented here is a pixel classifier based on four texture energy measures associated with each pixel in the image. An automated clustering procedure is used to label each pixel in the image with the label of its most probable class. The segmented images produced as the result of applying the algorithm to an example image are presented and discussed. The automated segmentation algorithm has been found to hold promise as an automated segmentation method.

Adult↗