Effects of a 99mTc-labeled murine immunoglobulin M antibody to CD15 antigens on human granulocyte membranes in healthy volunteers.

UNLABELLED: An injectible, 99mTc-labeled, murine immunoglobulin M antibody to stage-specific embryonic antigen-1 has been developed that can localize infections by binding to CD15 glycoproteins expressed on the cell membranes of human granulocytes in vivo after systemic administration. The purpose of this study was to measure its clinical effects on healthy people. METHODS: Multiple blood samples were aspirated before and after the intravenous administration of about 125 microg antibody labeled with approximately 370 MBq (10.0 mCi) 99mTc in 10 healthy human volunteers. Complete blood cell counts were performed at each time point. Whole-body scans were acquired contemporaneously with a dual-head gamma camera. The fraction of the administered dose at each time point was quantified in 18 regions of interest. Statistical analyses included paired t tests. RESULTS: Administration was associated with a transient decrease in the concentration of red and white blood cells in the whole blood. The effect always began within 3 min of administration. Its nadir was always reached 15-20 min after administration. There was full recovery with mild overcompensation in about an hour. The hematocrit dropped by a mean of 3.8% (P<0.002), whereas the total white blood cell count fell 44.0%+/-3.1% (P<0.001). The effect was most pronounced on the number of circulating granulocytes, which fell from 5.7+/-2.1 to 3.2+/-1.3x10(3)/microL blood. The drop paralleled a decrease in the percentage of whole blood radioactivity bound to the white blood cell membranes, which peaked at 50.4%+/-7.6% at 3 min after injection and then fell to 26.1%+/-9.3% over the next 30+/-13.4 min before recovering to 40.7%+/-8.2% at 2 h. Image analysis showed that the effect was temporally associated with an increase in the amount of radioactivity within the liver and the spleen. Recovery was associated with a decrease in hepatosplenic radioactivity. No evidence of cell destruction or agglutination could be detected. CONCLUSION: This study confirmed that administration of this radiolabeled antibody is associated with a transient decrease in the number of circulating granulocytes. However, there also seems to be a secondary hemodilutionlike effect on all blood components that has not been reported previously. The effect appears to be clinically silent and very short-lived.

Application of backpropagation neural networks to diagnosis of breast and ovarian cancer.

Neural network programs have been developed in an attempt to improve the diagnosis of breast and ovarian cancer using a group of laboratory tests and the age of the patient. The laboratory tests employed in this study include albumin, cholesterol, HDL-cholesterol, triglyceride, apolipoproteins A1 and B, NMR linewidth (the Fossel Index) and a tumor marker (i.e., CA 15-3 or CA 125). The breast cancer study involved 104 patients (45 malignant and 59 benign subjects). The ovarian cancer study involved 98 individuals (35 malignant, 36 benign and 27 control subjects). Methods are outlined for identification of the most influential input parameters and optimization of network structure and training. Network characteristics were contrasted with the test results of the appropriate serum tumor marker assay. For the breast cancer study, the best neural network program, using six input parameters, had a sensitivity of only 55.6% and a specificity of 72.9%. The tumor marker CA 15-3 alone gave results of 61.3% and 64.4%, respectively. For the ovarian cancer study, the best neural network program, using six input parameters, had a sensitivity of 80.6% and a specificity of 85.5%. The tumor marker CA 125 alone gave results of 77.8% and 82.3%, respectively. These methods provide an objective approach to neural network optimization and parameter selection applicable to other data bases of clinical and laboratory data.

Factors involved in the determination of triglycerides in serum: an international study.

Ten laboratories analysed five different specimens in duplicate on ten separate occasions by one, or several, of five common triglyceride methods. Simple statistical data are presented and, as far as possible, these are interpreted in the light of the methods used and the results of chemical analyses of the materials. Great variability was found between the results of the participating laboratories. The major factor involved seems to be the material specific nature of the methods under study. At least one method in common use is contraindicated.

The influence of drugs and disease activity on biochemical and haematological data in rheumatoid arthritis.

Biochemical and haematological data from 218 patients with rheumatoid arthritis were analysed and compared with data from a reference hospital population. The comparison demonstrated significant differences in several biochemical and haematological tests and that the patterns of change are different between males and females. The data were also analysed by conventional statistical methods and discriminant analysis using a computer to establish which tests were most influenced by the activity of the disease and drug therapy. The results obtained demonstrate marked difference between groups of patients with different disease activity or receiving different drugs. The discriminant analysis also identified those tests which differentiate these groups most effectively.

Analysis of ligase chain reaction products amplified in a silicon-glass chip using capillary electrophoresis.

Ligase chain reaction (LCR) is a useful molecular technique for detecting known point mutations. We report the first example of the use of a disposable silicon-glass micro-chip for LCR and the first application of capillary electrophoresis (CE) to analyze samples amplified by LCR in a chip. Silicon-glass chips were manufactured using conventional photolithography and anodic bonding. The chips provide three distinct advantages for LCR: excellent thermal conductivity, a micro reaction volume ( < 10 microliters), and reproducible, low-cost manufacturing. Investigation and quantitation of amplification efficiency of LCR in a chip or in a tube requires an analytical technique that is faster and more convenient than the conventional slab gel methods. Slab gel electrophoresis uses relatively large amounts of sample and is labor-intensive and time-consuming, and thus is unsuitable for the separation and detection of LCR products. In contrast CE requires sample volume (original LCR products) of less than 1 microliter and is therefore well-suited to analysis of the micro-volume reaction mixture from chips. We combined CE with a sensitive laser induced fluorescence (LIF) detection system for the rapid separation and quantitative detection of LCR products amplified from the lacI gene in a silicon-glass chip. Comparative studies were made with LCR between tubes and silicon-glass chips. CE-LIF analysis is ideally suited to examination of micro-LCR amplification with high throughput. The technologies may find medical uses in disease diagnosis and research.

Microchip-based Devices for Molecular Diagnosis of Genetic Diseases.

Microchips, constructed with a variety of microfabrication technologies (photolithography, micropatterning, microjet printing, light-directed chemical synthesis, laser stereochemical etching, and microcontact printing) are being applied to molecular biology. The new microchip-based analytical devices promise to solve the analytical problems faced by many molecular biologists (eg, contamination, low throughput, and high cost). They may revolutionize molecular biology and its application in clinical medicine, forensic science, and environmental monitoring. A typical biochemical analysis involves three main steps: (1) sample preparation, (2) biochemical reaction, and (3) detection (either separation or hybridization may be involved) accompanied by data acquisition and interpretation. The construction of a miniturized analyzer will therefore necessarily entail the miniaturization and integration of all three of these processes. The literature related to the miniaturization of these three processes indicates that the greatest emphasis so far is on the investigation and development of methods for the detection of nucleic acid, followed by the optimization of a biochemical reaction, such as the polymerase chain reaction. The first step involving sample preparation has received little attention. In this review the state of the art of, microchip-based, miniaturized analytical processes (eg, sample preparation, biochemical reaction, and detection of products) are outlined and the applications of microchip-based devices in the molecular diagnosis of genetic diseases are discussed.

An evaluation of the Greiner Selective Analyzer and its role in the clinical laboratory.

The Greiner Selective Analyzer (GSA II) was evaluated over a period of six months. The evaluation assessed the reliability, accuracy, and precision of the analyser for six determinations. The methods evaluated were for glucose, urea, creatinine, total protein, total bilirubin, and alkaline phosphatase. Comparison of results was also made with those obtained for the same specimens using the Technicon SMA 12160 Analyzer. Correlation and comparison of results indicate that the Greiner Selective Analyzer performed better for three of the methods but worse for serum creatinine determination. The role of the analyser as a routine tool in t heclinical laboratory was also evaluated during analyses of approximately 900 patient specimens. Other features evaluated were analytical range of the six methods under study, the economics of operation, temperature control, and electrical and mechanical safety. Additional key phrases: Accuracy, carryover, economics of operation, analytical ranges, reliability, precision.

Effect of paracetamol on uric acid determination.

The effect of paracetamol on uric acid determination using a phosphotungstate reduction method has been investigated. The results show that apparently raised values of serum uric acid may be reported in patients receiving therapeutic levels of this drug. Levels of serum uric acid were determined over a five-hour period in three patients who had received therapeutic doses of paracetamol. The assays of serum uric acid were performed by a method based on phosphotungstate reduction and by one employing uricase. The results illustrate the problems of using the former method for clinical purposes.

Enhanced chemiluminescence determination of alpha-amylase by use of amylose labelled with horseradish peroxidase.

An enhanced chemiluminescence method for determining soluble and immobilized alpha-amylase has been developed, based on use of an insoluble amylose substrate labelled with horseradish peroxidase. Soluble peroxidase-labelled fragments of the substrate, released by the action of alpha-amylase, are quantified by the peroxidase-catalyzed luminol-peroxide-p-iodophenol reaction. The detection limit for alpha-amylase was 125 fmole (12.5 nM). An insoluble amylopectin labelled with horseradish peroxidase was also effective as a substrate for this type of assay.

The application of backpropagation neural networks to problems in pathology and laboratory medicine.

Neural networks are a group of computer-based pattern recognition technologies that have been applied to problems in clinical diagnosis. This review focuses on one member of the group of neural networks, the backpropagation network. The steps in creating a backpropagation network are (1) collecting adequate training facts, (2) choosing the specific network structure, (3) training the network, and (4) cross-validating the trained network. The first published applications of backpropagation networks to problems in pathology and laboratory medicine have appeared recently. These applications are in the areas of image analysis and interpretation of laboratory results, and they demonstrate the feasibility of the approach.

Micromachined analytical devices: microchips for semen testing.

Micromachined devices (microchips) have been designed and tested for a range of clinically important assays. In this study we compare sperm motility determined using disposable glass microchips and a conventional Makler chamber. The 17 x 14 mm glass microchips contained three etched test structures each comprising either duplicate or quadruplicate analytical microchannels. Semen samples with sperm counts ranging from 21 to 78 million sperm per ml and forward progression scores of from 1+ to 3+ were evaluated and swimming times ranging from 360 s (3.3+ progression) to 770 s (1+,2 forward progression) observed in the microchips. Motility determined by the time taken for sperm to swim to the end of a microchannel (100 microns wide x 40 microns deep x 10 mm long) in the microchip correlated with forward progression of the sperm determined by the conventional Makler chamber method. This study demonstrates the feasibility of microchips for sperm motility testing and suggests that this technique would be applicable to the study of other types of motile cells.

Salt-induced swelling of meat: The effect of storage time, pH, ion-type and concentration.

The effect of hypertonic salt solutions on meat fibres has been studied as a function of post-mortem storage. Rabbit longissimus dorsi fibres (after about 20 h post-mortem storage at 4°C), were found to swell in hypertonic salt solutions, such as 0·25m KI or 0·6m KCl, to two to three times their original diameter. This swelling occurred in the fibre transverse plane only. X-ray diffraction shows swelling occurs by a combination of an increase in the myofilament lattice spacing and a loss of myofilament order. Fibre swelling is highly co-operative. At pHs below the isoelectric point of the myofibrillar proteins, hypertonic salt solutions induce fibre shrinkage. The post-mortem time course of response shows a peak at 18-35 h bounded by periods of minimal or zero response. We propose that the collagenous endomysial sheath acts as a restraint to myofibril swelling and that the characteristics of post-mortem swelling are a balance of the myofibrillar propensity to swell and the constraint of the endomysium.

Interaction between corticosteroids and beta-agonists in asthma.

It is often assumed that there is synergy between corticosteroids and beta-agonists, the two drugs most commonly used to treat asthma. A review of clinical studies of the effect of corticosteroids on the response to a beta-agonist finds no evidence to support this assumption except in subjects taking high doses of beta-agonists and with diminished responsiveness to a beta-agonist. There is some evidence that regular beta-agonist therapy may cause a temporary reduction in the efficacy of corticosteroids in asthma, an effect that is seen when the response to the beta-agonist has worn off. Further studies of this effect are required in view of the very widespread concomitant use of the two drugs.

Effect of long-term treatment with salmeterol on asthma control: a double blind, randomised crossover study.

OBJECTIVES: To determine the effect of adding salmeterol 50 micrograms twice daily for six months to current treatment in subjects with asthma who control their inhaled corticosteroid dose according to a management plan. DESIGN: A double blind, randomised crossover study. SETTING: Nottingham. SUBJECTS: 101 subjects with mild or moderate asthma taking at least 200 micrograms twice daily of beclomethasone dipropionate or budesonide. INTERVENTIONS: Salmeterol 50 micrograms twice daily and placebo for six months each, with a one month washout. Subjects adjusted inhaled steroid dose according to guidelines. MAIN OUTCOME MEASURE: Reduction in inhaled steroid use, exacerbations of asthma, and use of oral steroids. RESULTS: Data were available for 87 subjects. When compared with placebo salmeterol treatment was associated with a 17% reduction in inhaled steroid use (95% confidence interval 12% to 22%) with no significant difference in the number of subjects who had an exacerbation (placebo 25%, salmeterol 16%) or use of oral steroids. For secondary end points salmeterol treatment was associated with higher morning and evening peak expiratory flow and forced expiratory volume in one second; a reduction in symptoms, bronchodilator use and airway responsiveness to methacholine; and no effect on serum potassium concentration, 24 hour heart rate, or the final forced expiratory volume in one second achieved during a salbutamol dose-response study. CONCLUSIONS: In subjects who adjusted their inhaled steroid treatment according to guidelines the addition of salmeterol 50 micrograms twice daily was associated with a reduction in inhaled steroid use and improved lung function and symptom control.