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PubMed1983

Quantitation of a highly immunogenic leukocyte antigen (L1) by radioimmunoassay: methodological evaluation.

This paper describes a radioimmunoassay (RIA) for the quantitation of a major protein antigen (L1) released from human granulocytes. A protein A-carrying strain of Staphylococcus aureus was used as a solid phase IgG-binding reagent for the separation of antigen-antibody complexes from free antigen in the assay. A two-step version of the RIA may be completed in 2.5 h and has a detection limit of about 5 ng/ml. The sensitivity may be increased by minor modifications. A single step version takes 1.5 h and has adequate sensitivity to cover clinically important plasma levels. Selection and optimal use of reagents for the two-step procedure is described and their storage discussed. The simplicity of the procedure combined with high sensitivity and satisfactory accuracy, precision and reproducibility makes this assay suitable for quantitation of L1 in various body fluids.

Animals↗
PubMed1986

Cerebral responses evoked by stimulation of vesico-urethral junction in man: methodological evaluation of monopolar stimulation.

Cerebral responses after bipolar stimulation (BpS) and monopolar stimulation (MpS) of the vesico-urethral junction (VUJ), as well as skin surface stimulation of various hip and pelvic structures, were recorded in 13 normal control subjects. BpS of VUJ produced cerebral evoked potentials (CEPs) with a negative peak (mean latency 88.69 +/- 13.73 msec) following a smaller positive deflection, while MpS of the same sites with indifferent electrodes placed on various pelvic or hip structures resulted in different response latencies that were all significantly shorter than those of BpS. Shape and latency of responses after MpS of VUJ were similar to those elicited by skin surface stimulation of the abdominal wall and the iliac crest; those elicited by pudendal nerve stimulation at the glans were also similar in shape but significantly greater in latency. Results strongly indicate that during MpS of VUJ the somatic or muscle afferents located in the hip and pelvic region are stimulated; consequently, BpS technique should be considered the preferred technique for investigating the physiological condition of visceral afferents arising from VUJ.

Adult↗
PubMed1987

Alcohol treatment outcome evaluation methodology: state of the art 1980-1984.

The methodology of alcohol treatment outcome studies published during two sequential intervals from 1976 through mid-1984 is critically reviewed. Although considerable methodological improvements have occurred over time, major methodological deficiencies continue to characterize much of the literature, with inadequate reporting of subjects' pretreatment characteristics (e.g., severity of dependence) being the most striking problem. Pervasive differences across studies regarding the types of data gathered and the ways in which findings are reported seriously impede attempts to compare studies and weaken the types of conclusions that can be drawn about treatment efficacy in general. It is suggested that journal editors establish standards of reporting for follow-up studies.

Alcoholism↗
PubMed1987

Cochlear blood flow in the rat. A methodological evaluation of the microsphere method.

With the microsphere method it is possible to quantify the blood flow in various organs. The blood flow in the cochlea is only a very small part of the cardiac output and only relatively few microspheres are caught in this organ, which necessitates large groups of animals for such studies. The method has, however, not been fully evaluated for studies of small organs in small animals. In this study, 130 rats of various ages with normal or arterial hypertension were investigated. The blood flows of 97 animals were possible to evaluate. It was found that physiological parameters, such as PCO3, pH, PO2 and mean arterial blood pressure within the rather wide limits usually present in the anesthetized animal did not affect the cochlear blood flow to any great extent and that the method is feasible for studies of the blood flow through the inner ear in small animals.

Animals↗
PubMed2005

A spatially-evaluated methodology for assessing risk to a population from contaminated land.

A methodology is proposed which combines quantitative probabilistic human health risk assessment and spatial statistical methods (geostatistics) to produce an assessment of risks to human health from exposure to contaminated land, in a manner which preserves the spatial distribution of risks and provides a measure of uncertainty in the assessment. Maps of soil contaminant levels, which incorporate uncertainty, are produced from sparse sample data using sequential indicator simulation. A real, age-stratified population is mapped across the contaminated area, and intake of soil contaminants by individuals is calculated probabilistically using an adaptation of the Contaminated Land Exposure Assessment (CLEA) model. An abundance of information is contained in results which can be interrogated at the population and individual level, and mapped to provide a powerful visual tool for risk managers, enabling efficient targeting of risk reduction measures to different locations.

Environmental Exposure↗
PubMed1997

Multicenter study on the clinical value of fetal pulse oximetry. I. Methodologic evaluation. The French Study Group on Fetal Pulse Oximetry.

OBJECTIVE: Our purpose was to evaluate the feasibility of intrapartum fetal pulse oximetry, the distribution of fetal oxygen saturation values, and the relationship with the neonatal outcome in a population with an abnormal fetal heart rate. STUDY DESIGN: A prospective multicenter observational study was performed from June 1994 to November 1995. Fetal oxygen saturation was continuously recorded with use of a Nellcor N-400 fetal pulse oximeter in case of an abnormal fetal heart rate during labor. Simultaneous readings of fetal oxygen saturation and fetal blood analysis were obtained at inclusion and before birth. Feasibility, adverse effects, distribution of fetal oxygen saturation values, and relationship with neonatal outcome were assessed. RESULTS: One hundred seventy-four patients were included. From 172 attempted sensor placements, the procedure was impossible in three cases and fetal oxygen saturation values were obtained in 164 cases (95.3%). Physicians considered sensor placement an easier task than an attempt at fetal blood analysis (easy in 87.5% vs 78.9% for fetal blood analysis, p = 0.03). The mean reliable signal time (+/- SD) was 64.7% +/- 32% during the first stage. There were no serious adverse effects in the study population. The mean fetal oxygen saturation during the first stage of labor was 42.2% +/- 8.0% (10th to 90th percentile range 30% to 53%). Fetal oxygen saturation was significantly correlated with scalp pH (r = 0.29, p = 0.01) but not with neonatal umbilical artery pH or gas values. There was a significant association between low fetal oxygen saturation (< 30%) and poor neonatal condition. CONCLUSION: The feasibility of fetal pulse oximetry is satisfactory in clinical practice. It is easy to use and provides a fair rate of recorded values, even in a population with suspicion of fetal distress. A low fetal oxygen saturation is significantly associated with an abnormal neonatal outcome.

Adult↗
PubMed1994

Assessment of coping with muscular dystrophy: a methodological evaluation.

There is no published research on coping with muscular dystrophy among adults. In the present study, two questionnaires, the Reaction to the Diagnosis of Cancer Questionnaire (RDCQ) and the Mental Adjustment to Cancer scale (MAC), were modified in order to measure coping with muscular dystrophy (MD). A total of 60 people (16-64 years) with diagnosed MD answered the questionnaires in two interviews including semi-structured questions. The replies to these questions were analysed by two independent judges on the basis of the RDCQ and MAC categories for coping. The purpose was twofold: to investigate if cancer-coping categories could be used for the classification of interview answers concerning coping with MD, and to gain knowledge about specific coping with MD. Analysis indicated that 82% of replies to semi-structured questions concerned with emotion/appraisal-focused coping with MD can be described by means of RDCQ and MAC categories. Eight new categories were developed to classify the remaining 18%: Anticipation, Creation of new life values, Minimization, Establishment of control over everyday life, Secretiveness, Fear, Social comparison and Coping with heredity. In addition, 997 replies were classified to represent problem-focused coping. The judges reached good agreement with respect to the proportions of replies in the respective coping categories. However, kappa (kappa) values were within the range of fair to good agreement.

Adaptation, Psychological↗
PubMed2000

Quantitative bone scintigraphy. A methodological evaluation in prostate cancer.

PURPOSE: To evaluate a simple method for quantification of focal activity in bone scintigraphy (BS). MATERIAL AND METHODS: The gamma camera was calibrated using a phantom. Quantitative bone scintigraphy (QBS) was performed on 11 men recently diagnosed with prostate cancer (PCa), for whom routine BS showed involvement of the skeleton. Following endocrine therapy for 4 to 8 months, a second QBS was performed. Changes in QBS values were then compared to changes in serum levels of prostate-specific antigen (PSA). RESULTS: PSA response indicating regression of PCa was accompanied by a decrease in the QBS value in 8 of the 11 patients. The overall mean error of the QBS values was 15%. CONCLUSION: QBS according to this method is a relatively simple procedure that might contribute to objective evaluation of therapeutic effects in skeletal metastases, although its validity must be tested in a larger clinical material.

Aged↗
PubMed2005

Cerebral microdialysis methodology--evaluation of 20 kDa and 100 kDa catheters.

Microdialysis monitoring of cerebral metabolism is now performed in several neuro-intensive care units. Conventional microdialysis utilizes CMA70 catheters with 20 kDa molecular weight cut-off membranes enabling the measurement of small molecules such as glucose, lactate, pyruvate and glutamate. The CMA71 100 kDa molecular weight cut-off microdialysis catheter has recently been introduced to allow detection of larger molecules such as cytokines. The objective of this study was to perform in vitro and in vivo testing of the CMA71 microdialysis catheter, comparing its performance with the CMA70. In vitro comparison studies of three of each catheter using reference analyte solutions, demonstrated equivalent recovery for glucose, lactate, pyruvate and glutamate (range 94-97% for CMA70 and 88-103% for CMA71). In vivo comparison involved intracranial placement of paired CMA70 and CMA71 catheters (through the same cranial access device) in six patients with severe traumatic brain injury. Both catheters were perfused with CNS Perfusion Fluid without dextran at 0.3 microl min-1 with hourly sampling and bedside analysis on a CMA600 microdialysis analyser. The two catheters yielded equivalent results for glucose, lactate, pyruvate, glutamate and lactate/pyruvate ratio. CMA71 microdialysis catheters can, therefore, be used for routine clinical monitoring of extracellular substances, as well as for their intended research role of larger molecular weight protein sampling.

Adolescent↗
PubMed2004

Methodological evaluation to analyze functional recovery after sciatic nerve injury.

The Basso, Bresnahan and Beattie (BBB) locomotor scale has not been tested to evaluate functional consequences of peripheral nerve lesions. Alternative methods to evaluate animal functional recovery after sciatic nerve injury are desirable. Male Wistar rats had a right sciatic nerve segment exposed and were divided in three experimental groups: Sham (wound open, 10 min), Sham-device (nerve segment between crushing device, 10 min), and Crush-force (nerve crushing load of 15,000 g/1,000 mm Hg/mm(2), 10 min). Animals were evaluated preoperatively, 1, 7, 14, 21, and 28 days after procedure by calculation of Sciatic Functional Index (SFI), BBB score and open arena exploratory activity. The primary findings of the present study were (1) the SFI calculated by either DeMedinaceli, Carlton and Goldberg, and Bain formulae were highly correlated; (2) the BBB score evaluation was highly correlated with the SFI; (3) the BBB motor scale was able to detect functional impairments not recognized by the SFI; and (4) open arena exploratory activity was a poor method to detect sciatic nerve impairment. In conclusion, the BBB prescribed functional deficits on the sham-device and crush-force groups even when the SFI indicated full recovery. This greater sensitivity may prove useful when comparing new therapeutic approaches to nerve regeneration.

Animals↗
PubMed1987

Outward Na-K-Cl cotransport in human red blood cells: a methodological evaluation of method for epidemiologic investigations.

The present report evaluates a method for the determination of the maximal velocity of the outward sodium, potassium, chlorine (Na-K-Cl) cotransport in erythrocytes. The loading procedure was carried out using the ionophore nystatin. The technical error for this assay, based on concurrent analysis of randomized blind duplicate samples, was 8.04% of the sample mean and 27.5% of the sample standard deviation. The assay was stable over time and reproducible. Fasting was not necessary. The ratio of intra- to interindividual variances was small. The cells can be safely preserved in an appropriate preservation solution without modification of the ion transport assay. The assay appears to be adequate for use in large-scale field trials.

Chlorides↗
PubMedDate not supplied

Immune complexes in experimental syphilis: a methodologic evaluation.

Sustained antigenic stimulation following intravenous infection of rabbits with Treponema pallidum results in antibody responses and the formation of immune complexes. Complexes in greater than 300 sera from normal rabbits and from animals with experimental syphilis were measured by Raji cell and polyethylene glycol-complement consumption assays. Profound increases in both the incidence and levels of immune complexes were seen between the third and seventh weeks of infection. Single radial immunodiffusion studies revealed moderate increases in immunoglobulin levels and depression of C3 levels during the third and fourth weeks of infection. Penicillin treatment was followed by normalization of C3 levels and clearance of immune complexes. Lower levels of complexes in the sera of intradermally infected rabbits and the absence of immune complexes in intratesticularly infected animals suggest that immune complexes in the circulation may be related to the extent and severity of cutaneous involvement.

Animals↗
PubMed1980

The influence of serum on random migration and chemotaxis of polymorphonuclear leucocytes: methodological evaluation using sera from infection-prone patients and normals.

The effects on normal polymorphonuclear leucocytes (PMN) of chemotactic and chemokinetic factors in sera from normal subjects and infection-prone patients were examined by means of the leading-front technique, using a modified Boyden chamber. The analysis scheme used made it possible to differentiate between chemotactic and chemokinetic factors and demonstrated that different factors account for the major chemotactic and chemokinetic activities of serum. The major chemotactic activity of unactivated serum was heat-labile, and the chemotactic activity of factor XII-deficient sera was normal, suggesting the major chemotactic activity to be distinct from C5a and factor XII-dependent pathways. The existence of both heat-stable and heat-labile chemokinetic factors was shown. The possibility that the reduced chemokinetic effect of several patient sera was caused by abnormal levels of the serum proteins alpha 1-antitrypsin, alpha 2-macroglobulin and albumin was excluded.

Adolescent↗
PubMed1993

Methodological evaluation of indirect calorimetry data in lean and obese rats.

1. The applicability of current indirect calorimetry formulae to the study of energy and substrate balances on obese rats has been evaluated. The energy consumption of series of 60-day rats of Wistar, lean and obese Zucker stock were studied by means of direct and indirect calorimetry, and by establishing their energy balance through measurement of food intake and retention. Calorimetric studies encompassed a 24 h period, with gas and heat output measurements every 2 or 5 min, respectively, for direct and indirect calorimetry. 2. The analysis of fat composition (diet, whole rat, and synthesized and oxidized fat) showed only small variations that had only a limited effect on the overall energy equation parameters. 3. A gap in the nitrogen balance, which represents a urinary N excretion lower than the actual protein oxidized, resulted in significant deviations in the estimation of carbohydrate and lipid oxidized when using the equations currently available for indirect calorimetry. 4. Analysis of the amino acid composition of diet and rat protein as well as of the portion actually oxidized, and correcting for the nitrogen gap allowed the establishment of a set of equations that gave better coincidence of the calculated data with the measured substrate balance. 5. The measured heat output of all rats was lower than the estimated values calculated by means of either indirect calorimetry of direct energy balance measurement; the difference corresponded to the energy lost in water evaporation, and was in the range of one-fifth of total energy produced in the three rat stocks. 6. Wistar rats showed a biphasic circadian rhythm of substrate utilization, with alternate lipid synthesis/degradation that reversed that of carbohydrate, concordant with nocturnal feeding habits. Zucker rats did not show this rhythm; obese rats synthesized large amounts of fat during most of the light period, consuming fat at the end of the dark period, which suggests more diurnal feeding habits. Lean Zucker rats showed a similar, but less marked pattern. 7. The results obtained indicate that lean and obese rats can be studied using the same indirect calorimetry formulae provided that there is an adequate measure of protein oxidation and the composition of diet does not differ.

Amino Acids↗
PubMed2000

Multisite comparison of CD4 and CD8 T-lymphocyte counting by single- versus multiple-platform methodologies: evaluation of Beckman Coulter flow-count fluorospheres and the tetraONE system. The NIAID DAIDS New Technologies Evaluation Group.

New analytic methods that permit absolute CD4 and CD8 T-cell determinations to be performed entirely on the flow cytometer have the potential for improving assay precision and accuracy. In a multisite trial, we compared two different single-platform assay methods with a predicate two-color assay in which the absolute lymphocyte count was derived by conventional hematology. A two-color method employing lymphocyte light scatter gating and Beckman Coulter Flow-Count fluorospheres for absolute counting produced within-laboratory precision equivalent to that of the two-color predicate method, as measured by coefficient of variation of replicate measurements. The fully automated Beckman Coulter tetraONE System four-color assay employing CD45 lymphocyte gating, automated analysis, and absolute counting by fluorospheres resulted in a small but significant improvement in the within-laboratory precision of CD4 and CD8 cell counts and percentages suggesting that the CD45 lymphocyte gating and automated analysis might have contributed to the improved performance. Both the two-color method employing Flow-Count fluorospheres and the four-color tetraONE System provided significant and substantial improvements in between-laboratory precision of absolute counts. In some laboratories, absolute counts obtained by the single-platform methods showed small but consistent differences relative to the predicate method. Comparison of each laboratory's absolute counts with the five-laboratory median value suggested that these differences resulted from a bias in the absolute lymphocyte count obtained from the hematology instrument in some laboratories. These results demonstrate the potential for single-platform assay methods to improve within-laboratory and between-laboratory precision of CD4 and CD8 T-cell determinations compared with conventional assay methods.

CD4-CD8 Ratio↗
PubMed1976

Antibacterial susceptibility testing by the dip-slide technique: a methodological evaluation.

Antibacterial susceptibility testing of urine specimens by the dip-slide method was compared to the standardized disc diffusion method. A boarder line of 12 mm inhibition zone on the slide could be used to select strains resistant to sulphisadimidine, ampicillin, nitrofurantoin or nalidixic acid. In the case of the sulphonamide, the results were grewtly influenced by composition of the medium. Susceptibility testing with the dip-slide method seems to be a useful test to differentiate strains likely to respond to therapy from resistant strains.

Ampicillin↗
PubMedDate not supplied

Filter immunocytology: methodologic evaluation of a new assay for the diagnosis of urothelial cancer.

OBJECTIVE: Several investigators have demonstrated the high sensitivity of immunocytology in the diagnosis of transitional cell carcinoma (TCC). A new technique, designated "filter immunocytology" (FLIC), simplifies the technique of quantitative immunocytology, considerable decreases assay time and increases the percentage of assessable specimens. STUDY DESIGN: Voided urine samples were obtained from 89 patients without evidence of TCC and from 91 patients with histologically proven TCC. The cells were transferred onto a polycarbonate membrane. Immunostaining was performed using monoclonal antibody. Due ABC 3, directed against a differentiation antigen on urothelial cells. Specimens containing > 35% positive urothelial cells were regarded as abnormal. RESULTS: Of 153 specimens 180 (85%) were assessable. The investigation of 76 specimens from control patients and 77 from patients with TCC yielded a specificity of 86% and a sensitivity of 75%, respectively. Sensitivity did not correlate with tumor grade. Despite high interobserver and intrapatient variations regarding the amount of antigen-positive cells, a concordant attribution to either "normal" or "abnormal" was made in > 95% of cases. Intraobserver variation was small and did not influence the test result. CONCLUSION: These results suggest that FLIC assay may be a valuable adjunct to conventional cytology. A careful prospective investigation appears to be worthwhile to further define the indications for this technique.

Antibodies, Monoclonal↗