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At least 1,243 records · Page 69Linked to original sources

Advantage of buffered solutions or automated capnometry in air-filled balloons for use in gastric tonometry.

OBJECTIVE: To test accuracy, reproducibility and time constants of pCO2 measurement with the tonometric technique, using different media for filling the silastic balloon (saline, phosphate buffer, citrate buffer, air) and employing different analyzer devices (ABL3, ABL330, Nova Stat 5, automated capnometry). DESIGN: Comparative laboratory study of different tonometric techniques, measuring test solutions with known pCO2 values due to pre-equilibration with three different pCO2 concentrations. SETTING: Clinical laboratory of a university hospital intensive care unit. MEASUREMENTS AND RESULTS: The use of saline, as suggested for routine tonometry, led to negative bias values throughout, i.e. underestimation of pCO2 values, the extent of which depended on the blood gas analyzer device employed. Registration of the equilibration kinetics showed that full equilibration demanded 90 min regardless of the environmental pCO2 level. Replacing saline by buffered electrolyte solutions resulted in a significant improvement of bias, but did not change the kinetics of pCO2 equilibration. The employment of air-filled balloons, combined with automated capnometry, led to very low bias values, approaching zero, for all pCO2 levels, along with excellent precision. Time constants of equilibration were dramatically reduced, with full equilibration being achieved within 12.5 min. CONCLUSIONS: Buffered electrolyte solutions are preferable to saline for achieving reliable pCO2 measurements in gastric tonometry. Air-filled balloons, combined with automated capnometry, present excellent accuracy and reproducibility together with short equilibration times, thus offering "on-line" monitoring of even rapid changes in environmental pCO2.

Bias↗

Automated in situ trace element analysis of silicate materials by laser ablation inductively coupled plasma mass spectrometry.

This paper describes the automated in situ trace element analysis of solid materials by laser ablation (LA) inductively coupled plasma mass spectrometry (ICP-MS). A compact computer-controlled solid state Nd:YAG Merchantek EO UV laser ablation (LA) system has been coupled with the high sensitivity VG PQII S ICP-MS. A two-directional communication was interfaced in-house between the ICP-MS and the LA via serial RS-232 port. Each LA-ICP-MS analysis at a defined point includes a 60 s pre-ablation delay, a 60 s ablation, and a 90 s flush delay. The execution of each defined time setting by LA was corresponding to the ICP-MS data acquisition allowing samples to be run in automated cycle sequences like solution auto-sampler ICP-MS analysis. Each analytical cycle consists of four standards, one control reference material, and 15 samples, and requires about 70 min. Data produced by Time Resolved Analysis (TRA) from ICP-MS were later reduced off-line by in-house written software. Twenty-two trace elements from four reference materials (NIST SRM 613, and fused glass chips of BCR-2, SY-4, and G-2) were determined by the automated LA-ICP-MS method. NIST SRM 610 or NIST SRM 613 was used as an external calibration standard, and Ca as an internal standard to correct for drift, differences in transport efficiency and sampling yield. Except for Zr and Hf in G-2, relative standard deviations for all other elements are less than 10%. Results compare well with the data reported from literature with average limits of detection from 1 ng x g(-1) to 455 ng x g(-1) and less than 100 ng x g(-1) for most trace elements.

Journal Article↗

Automated cardiac auscultation for detection of pathologic heart murmurs.

Experienced cardiologists can usually recognize pathologic heart murmurs with high sensitivity and specificity, although nonspecialists with less clinical experience may have more difficulty. Harsh, pansystolic murmurs of intensity grade > or = 3 at the left upper sternal border (LUSB) are likely to be associated with pathology. In this study, we designed a system for automatically detecting systolic murmurs due to a variety of conditions and examined the correlation between relative murmur intensity and likelihood of pathology. Cardiac auscultatory examinations of 194 children and young adults were recorded, digitized, and stored along with corresponding echocardiographic diagnoses, and automated spectral analysis using continuous wavelet transforms was performed. Patients without heart disease and either no murmur or an innocent murmur (n = 95) were compared to patients with a variety of cardiac diagnoses and a pathologic systolic murmur present at the LUSB (n = 99). The sensitivity and specificity of the automated system for detecting pathologic murmurs with intensity grade > or = 2 were both 96%, and for grade > or = 3 murmurs they were 100%. Automated cardiac auscultation and interpretation may be useful as a diagnostic aid to support clinical decision making.

Adolescent↗

Automated generation and evaluation of specific MHC binding predictive tools: ARB matrix applications.

Prediction of which peptides can bind major histocompatibility complex (MHC) molecules is commonly used to assist in the identification of T cell epitopes. However, because of the large numbers of different MHC molecules of interest, each associated with different predictive tools, tool generation and evaluation can be a very resource intensive task. A methodology commonly used to predict MHC binding affinity is the matrix or linear coefficients method. Herein, we described Average Relative Binding (ARB) matrix methods that directly predict IC(50) values allowing combination of searches involving different peptide sizes and alleles into a single global prediction. A computer program was developed to automate the generation and evaluation of ARB predictive tools. Using an in-house MHC binding database, we generated a total of 85 and 13 MHC class I and class II matrices, respectively. Results from the automated evaluation of tool efficiency are presented. We anticipate that this automation framework will be generally applicable to the generation and evaluation of large numbers of MHC predictive methods and tools, and will be of value to centralize and rationalize the process of evaluation of MHC predictions. MHC binding predictions based on ARB matrices were made available at http://epitope.liai.org:8080/matrix web server.

Animals↗

Comparison of the classical manual pushed wedge films, with an improved automated method for making blood smears.

Comparison of apparatus for automatically spreading peripheral blood films (GENES) to manual wedge-pull technique has been performed, showing that the average size of optimal area for counting is twice larger in automated smears compared to manual ones. The variability of WBC repartition has been studied in doing differential count on manual and automated smears and both are compared to results obtain with an independent technique (Coulter STKS) for differential. The correlation coefficients for each WBC population show a closer relationship between automated smear preparations and reference instrument (STKS) than with manual spreading smears. Monocyte differential is particularly influenced by the variability of manual spreading procedures.

Blood Cell Count↗

Correlation between automated writing movements and striatal dopaminergic innervation in patients with Wilson's disease.

Handwriting defects are an early sign of motor impairment in patients with Wilson's disease. The basal ganglia being the primary site of copper accumulation in the brain suggests a correlation with lesions in the nigrostiatal dopaminergic system. We have analysed and correlated striatal dopaminergic innervation using [(123)I]beta-CIT-SPECT and automated handwriting movements in 37 patients with Wilson's disease. There was a significant correlation of putaminal dopaminergic innervation with fine motor ability (p < 0,05 for NIV [number of inversion in velocity], NIA [number of inversion in acceleration], frequency). These data suggest that loss of dorsolateral striatal dopaminergic innervation has a pathophysiological function for decreased automated motor control in Wilson's disease. Furthermore analysis of automated handwriting movements could be useful for therapy monitoring and evaluation of striatal dopaminergic innervation.

Adult↗

A novel method for patch-clamp automation.

An increasing demand of the pharmaceutical industry for automated electrophysiological stations for ion channel drug discovery has recently resulted in the development of several commercial platforms for secondary and safety screening of ion channel modulators. These commercial systems have demonstrated an enhanced throughput, however, often at the expense of some quality-sensitive aspects of traditional patch-clamp recordings. To improve data quality and content, we have developed a patch-clamp robot that fully automates manual patch-clamp recordings, including patch pipette handling, gigaseal formation, obtaining whole-cell or perforated-cell configuration, drug application, and data acquisition. Utilization of glass micropipettes results in high-quality electrophysiological recordings with an overall success rate of about 30% in perforated-cell mode. A fast drug application system with low volume requirements (1-1.5 ml) allows the study of ligand-gated ion channels on a millisecond scale. As proof-of-concept, we present two assays developed for voltage-gated human ether-a-go-go-related and ligand-gated alpha(7) nicotinic receptor ion channels. The system throughput was a single concentration-response curve every 30-40 min or 12-17 6-point concentration-response curves daily, representing a significant improvement of typical manual patch-clamp throughput. This system represents an efficient method for patch-clamp automation without the need for a complex and expensive electrophysiological set-up for cell visualization.

Animals↗

Automated fast perfusion of Xenopus oocytes for drug screening.

Fast ('concentration jump') applications of neurotransmitters are crucial for screening studies on ligand-gated ion channels. In this paper, we describe a method for automated fast perfusion of neurotransmitters (or drugs) during two-microelectrode voltage-clamp experiments on Xenopus oocytes. The oocytes are placed in a small bath chamber that is covered by a glass plate with two channels for the microelectrodes that are surrounded by a quartz funnel serving as a reservoir for test solutions. The oocytes are perfused in a vertical direction via the two channels in the plate. Automation of compound delivery is accomplished by means of a programmable pipetting workstation. A mean rise time for 10-90% current increase through muscle-type nACh channels of 55.0+/-1.3 ms (30 muM acetylcholine) was estimated. Automation, fast perfusion rates, and economical use of compounds ( approximately 100 mul/data point) make the system suitable for screening studies on ligand- and voltage-gated ion channels.

Animals↗

Rapid testing for nitrate reductase activity of Mycobacterium tuberculosis grown in an automated culture system.

In order to reduce the time to detection of nitrate reductase activity, which is arguably the most widely used phenotypic trait to differentiate between Mycobacterium tuberculosis, Mycobacterium bovis and Mycobacterium bovis BCG, the following study was conducted using cultures grown in an automated system. Automated culture systems, which are typically based on liquid medium, have greatly reduced the time-to-recovery of mycobacteria. Yet subsequent testing of isolates for nitrate reductase activity may take several weeks, because culture on solid media is required. Presented here is a procedure to obtain a final result within 24 h for nitrate reductase activity of cultures grown in an automated culture system. Using this procedure, Mycobacterium tuberculosis was rapidly differentiated from Mycobacterium bovis and Mycobacterium bovis BCG.

Antigens, Bacterial↗

PASA--a program for automated protein NMR backbone signal assignment by pattern-filtering approach.

We present a new program, PASA (Program for Automated Sequential Assignment), for assigning protein backbone resonances based on multidimensional heteronuclear NMR data. Distinct from existing programs, PASA emphasizes a per-residue-based pattern-filtering approach during the initial stage of the automated 13Calpha and/or 13Cbeta chemical shift matching. The pattern filter employs one or multiple constraints such as 13Calpha/Cbeta chemical shift ranges for different amino acid types and side-chain spin systems, which helps to rule out, in a stepwise fashion, improbable assignments as resulted from resonance degeneracy or missing signals. Such stepwise filtering approach substantially minimizes early false linkage problems that often propagate, amplify, and ultimately cause complication or combinatorial explosion of the automation process. Our program (http://www.lerner.ccf.org/moleccard/qin/) was tested on four representative small-large sized proteins with various degrees of resonance degeneracy and missing signals, and we show that PASA achieved the assignments efficiently and rapidly that are fully consistent with those obtained by laborious manual protocols. The results demonstrate that PASA may be a valuable tool for NMR-based structural analyses, genomics, and proteomics.

Algorithms↗

AUREMOL-RFAC-3D, combination of R-factors and their use for automated quality assessment of protein solution structures.

We present here the computer program AUREMOL-RFAC-3D that is a generalization of the previously published program RFAC for the fully automated estimation of residual indices (R-factors) from 2D NOESY spectra. It is part of the larger AUREMOL software package (www.auremol.de). RFAC-3D calculates R-factors directly from two-dimensional homonuclear NOESY spectra as well as from three-dimensional (15)N or (13)C edited NOESY-HSQC spectra and thus extends the application range to larger proteins. The fully automated method includes automated peak picking and integration, a Bayesian noise and artifact recognition and the use of the complete relaxation matrix formalism. To enhance the reliability of the calculated R-factors the method is also generalized to calculate combined R-factors from a set of 2D and 3D-spectra. For an optimal combination of the information derived from different sources a plausible formalism had to be derived. In addition, we present a novel direct R-factors based measure that correlates an R-factors as defined in this paper to the root mean square deviation of the actual structure from the optimal structure. The new program has been successfully tested on the histidine containing phosphocarrier protein (HPr) from Staphylococcus carnosus and on the Ras-binding domain (RBD) of the Ral guanine-nucleotide dissociation stimulation factor (RalGDS).

Bacterial Proteins↗

Methods and results for semi-automated cloning using integrated robotics.

The Joint Center for Structural Genomics (JCSG) has emphasized automation and parallel processing approaches. Here, we describe automated methods used across the cloning process with results from JCSG projects. The protocols for PCR, restriction digests and ligations, as well as for gel electrophoresis and microtiter plate assays have all been automated. The system has the capacity to routinely process 384 clones a week. This throughput can adequately supply our expression and purification pipeline with expression-ready clones, including novel targets and truncations. The utility of our system is demonstrated by our results from three diverse projects. In summary, 94% of the PCR amplicons generated to date have been successfully cloned and verified by sequencing (83% of the total attempted targets). Our results demonstrate the capabilities of this robotic platform to provide an avenue to high-throughput cloning which requires little manpower and is rapid and cost-effective while providing insights for method optimization.

Cloning, Molecular↗

Automated quality control in computed radiography.

PURPOSE: The purpose of this paper is to describe the automation of quality control procedures on photo-stimulable imaging plates by means of an image-processing tool providing automatic reading of the images and automatic calculation of the quality parameters monitored. MATERIALS AND METHODS: Quality-control procedures were performed according to the main available guidelines. The quality assurance programme was applied to several Kodak and Philips devices in four radiological departments. The automatic image-processing tool was developed using public domain software (Java-based ImageJ software) and contains both reading and computation procedures. RESULTS: The quality checks and algorithms described were successfully applied, proving useful for identification of defective plates and for implementation of the quality assurance programme. The use of automation allowed significant savings in the time required for quality checks. CONCLUSIONS: Completely automated image reading allows substantial economic and human resources savings, as it eliminates much of the transfer, reproduction, processing and filing procedures.

Algorithms↗

Evaluating the diagnosis and prognosis of hypertension by automated blood pressure monitoring: outline of a symposium.

The technique of automated whole-day blood pressure monitoring by noninvasive, lightweight portable devices has recently become available for clinical use. This article, together with the six that follow it, comprise the proceedings of a symposium on the diagnostic and prognostic use of this new approach. There is a growing belief that standard clinical methods may overdiagnose hypertension (the phenomenon of "white coat" or "office" hypertension), subjecting some patients to unnecessary long-term treatment. The automated blood pressure monitoring procedure may be especially helpful in evaluating patients whose office-diagnosed hypertension is not associated with confirmatory clinical findings or history. Data derived from whole-day blood pressure measurements appear to be more reproducible than conventional blood pressure values and correlate more closely with evidence for cardiovascular changes. This symposium has addressed some of the issues associated with the monitoring techniques, including the accuracy of values obtained with noninvasive equipment, interpretation of the data, and costs of the procedure. The evidence and arguments presented by these papers suggest that automated blood pressure monitoring is becoming a justified and desirable procedure in selected patients with a preliminary clinical diagnosis of mild to moderate hypertension.

Blood Pressure↗

Development and validation of an automated method of the Selvester QRS scoring system for myocardial infarct size.

The Selvester QRS score for estimation of myocardial infarct (MI) size from the standard 12-lead electrocardiogram (ECG) has not yet achieved wide recognition as a valuable tool in the routine assessment of the MI patient, primarily because of the practical limitations inherent to manual application. This study examined the ECGs of 438 patients (105 normal subjects, 161 with "possible" MI and 172 with "definite" MI based on data from cardiac catheterization) to develop software for an automated method of the Selvester system in attempts to overcome the manual constraints. After a comprehensive validation process involving extensive interactions between the manual scorer and the software developer, an automated method of the Selvester system was generated that had a high correlation with manual application (r = 0.94) and was superior regarding time, training, reader bias, reproducibility and precision of measurement. These results indicate that an automated version of the Selvester QRS scoring system would resolve many of the limitations of manual application and would provide a reliable, technically accurate estimate of MI size that could be incorporated into ECG diagnostic programs and used in standard digital ECG machines.

Cardiac Catheterization↗

Measurement of cardiac output by automated single-breath technique, and comparison with thermodilution and Fick methods in patients with cardiac disease.

Accurate noninvasive methods are needed for determination of cardiac output. Current methods are generally complex or may be unreliable. A previously described method, based on absorption of acetylene gas during a constant exhalation that enables calculation of cardiac output by estimating pulmonary capillary circulation, is incorporated in a new, automated commercial product (SensorMedics 2200). In this study, cardiac output by single-breath acetylene blood flow measured with this device was compared with the standard thermodilution and direct Fick methods in 20 patients undergoing cardiac or pulmonary artery catheterization. Patients inhaled test gas mixture to total lung capacity and exhaled at a constant rate through an adjustable resistor. Lung volumes and noninvasive acetylene blood flow value were calculated automatically. Correlation between the automated single-breath technique and both thermodilution and Fick cardiac output determinations was very high (correlation coefficients were 0.90 and 0.92, respectively), regression slopes were close to identity (0.98 and 0.90), and bias (-0.39 and -0.79 liter/min) and precision (0.94 and 1.02) were good; when shunt correction was applied, bias was reduced to 0.06 and 0.35 liter/min, respectively. Rapid, accurate, noninvasive measurement of cardiac output was easily obtained using the automated device. This technique may have a wide applicability for noninvasive evaluation of patients with cardiac disease and for monitoring effects of therapeutic interventions.

Absorption↗

On-line estimation of changes in left ventricular stroke volume by transesophageal echocardiographic automated border detection in patients undergoing coronary artery bypass grafting.

Echocardiographic automated border detection can determine the interface between blood and myocardial tissue and calculate left ventricular (LV) cavity area in real-time. The objective was to determine if on-line measurements of LV cavity area by transesophageal automated border detection could be used to determine beat-to-beat changes in stroke volume in humans. Studies were attempted on 9 consecutive patients, aged 66 +/- 8 years, undergoing coronary bypass surgery. Stroke volume was measured by electromagnetic flow from the ascending aorta, and LV cavity area was measured at the midventricular short-axis level. Simultaneous area and flow data were recorded on a computer workstation through a customized interface with the ultrasound system. Recordings were performed during baseline apnea and rapid alterations induced by inferior vena caval occlusions before and after cardiopulmonary bypass. Measurements of stroke area (maximal area-minimal area) were correlated with stroke volume for matched beats. Data were available for analysis on 8 of 9 patients before and on 5 patients after cardiopulmonary bypass for 644 beats. Stroke area was closely correlated with stroke volume both before (mean R = 0.94 +/- 0.03, SEE = 0.33 +/- 0.12 cm2) and after (mean R = 0.92 +/- 0.05, SEE = 0.59 +/- 0.81 cm2) cardiopulmonary bypass. The slopes of these stroke area-stroke volume relations were quite reproducible from before to after cardiopulmonary bypass in the same patient but varied between individual patients. Transesophageal automated border detection has potential for on-line estimation of changes in stroke volume in selected patients.

Aged↗

Automated methods for detection of bacteriuria.

Urine specimens represent a large portion of cultures received by the clinical microbiology laboratory. Much time and effort are spent screening these specimens and approximately 80 percent show no growth. The methods employed for detection of bacteriuria include microscopic, chemical, culture, and automated. The most widely used procedure has been an agar plate culture method. However, this method requires overnight incubation and therefore results are delayed. Within the last decade many automated methods have been introduced for detecting bacteriuria, and results are available more rapidly than with the agar plate culture method. In addition to decreased detection time, these systems are accurate and cost-effective. The purpose of this report is to review these automated bacteriuria screening systems, which include bioluminescence, colorimetry, electrochemical screening, electrical impedance, microcalorimetry, photometry, and radiometry. Accuracy, detection time, and cost are also discussed in this review.

Adenosine Triphosphate↗