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Detection of antibodies to Plasmodium falciparum sporozoites by employing synthetic peptides.

A new achievement in the immunodiagnosis of malaria has been reached after the knowledge of the molecular structure of some plasmodial antigens has become available. One example is given by the repetitive immunodominant epitope of Plasmodium falciparum circumsporozoite protein, which consists of 4 tandemly repeated aminoacids (Asn-Ala-Asn-Pro = NANP). A large synthetic peptide reproducing 40 NANP repeats, (NANP)40, has been shown to reproduce efficiently the native antigen in the CS protein and has been used to develop an enzyme-linked immunosorbent assay (ELISA) for the detection of antisporozoite antibodies in individuals from malaria-endemic countries. This (NANP)40 ELISA has been employed in a longitudinal study in a rural community in Tanzania. The results obtained have shown (i) that the presence of anti-(NANP) antibodies is associated with a certain degree of protective immunity; and (ii) that genetic factors could play a role in the host immune responsiveness to (NANP). Such an ELISA can be easily applied to field research and can be useful for monitoring the immune status of populations participating, in the future, to malaria vaccination trials employing P. falciparum sporozoite peptides.

Adolescent↗

Reproducibility of the 14C urea breath test repeated after 1 week.

OBJECTIVES: The aim of this study was to determine the numerical and categorical reproducibility of the 14C urea breath test in a routine clinical setting. BACKGROUND: The 14C urea breath test is used for diagnosis of Helicobacter pylori infection and is regarded by some centers as the gold standard. METHODS: In 140 referrals, duplicate tests were performed 1 wk apart. Eighteen patients were excluded because they had taken antibiotics or had changed medication that could influence the gastric urease activity between the tests. RESULTS: The limits of agreement between the repeated tests were 45-234%. Hence, the numerical result of a repeated test was 95% likely to differ from the first with a factor of approximately two or less. However, when patients were classified as "positive" or "negative" according to our previously determined cut-off limit, only three patients were discordantly classified by the two tests. The observed agreement between the tests was 98%, and the chance-corrected proportional agreement (kappa) was 95%, which is far better than that reported for the histological diagnosis of gastric H. pylori infection. CONCLUSION: The diagnostic reproducibility of the urea breath test was very good.

Adolescent↗

[Repeated use of Drug Infusion Balloon Catheter for postoperative pain management].

The purpose of this study was to evaluate repeated use of Drug Infusion Balloon catheter (DIBC) experimentally and clinically. 1. Experimental study: The changes in flow rate by repeated use of DIBC (40 ml.12 h-1 type) were measured under room temperature (21 degrees C). 2. Clinical study: Ten patients scheduled for upper abdominal surgery were studied. DIBC (40 ml.12 h-1 type) was filled with midazolam, buprenorphine, and lidocaine. Twelve hours later, the same DIBC was filled with buprenorphine and lidocaine. The mean flow rates were, on first use; 2.91 +/- 0.51, on the second use; 2.88 +/- 0.45, on the third use; 2.75 +/- 0.44, and on the fourth use; 2.69 +/- 0.38 (ml.h-1, mean +/- SD, n = 6, P < 0.05). In the clinical study, pain scores, rates of supplemental analgesic infusion during 0-12 h period were not different from those during 12-24 h period. No complications were observed in this study. Repeated use of DIBC up to four times yielded reproducible results (+/- 8%). Repeated use of DIBC for postoperative pain relief, changing the contents according to the severity of pain, might be a useful and cost saving method.

Adult↗

Ovine bronchoalveolar lavage cellularity: reproducibility and the effect of multiple repeated lavage.

We sought to determine the normal within-sheep variability of bronchoalveolar cellular parameters and to infer the numbers of sheep required to detect a defined significant change within the same. We further sought to examine the effect of repeated bronchoalveolar lavage (BAL) on those parameters. Neither the total cell number nor the absolute numbers of any given cell type changed significantly following repeated BAL. Additionally, there was no significant change in percentage differential cell populations over time in randomly sampled lobes. However for lobes sampled repeatedly, a significant change in the percentage of lymphocytes was detected. Although the percentages of neutrophils in repeatedly and randomly sampled lobes were significantly correlated, the percentage found in the former tended to be greater and more variable than in the latter. As a consequence, larger group sizes are required to detect given changes in neutrophil percentages in repeatedly sampled lobes over time when compared with detecting equivalent changes in randomly selected lobes.

Animals↗

A reproducible assay of polymerase chain reaction to detect trinucleotide repeat expansion of Huntington's disease and senile chorea.

A simple and reproducible method of polymerase chain reaction (PCR) assay was established to detect trinucleotide repeat expansion for Huntington's disease (HD) using a new DNA polymerase and buffer system. The system consists of an extremely heat stable DNA polymerase (Pfu), and a buffer supplemented with ammonium sulfate and dimethyl sulfoxide. Previous methods to amplify expanded alleles for HD have been very complex in PCR conditions, but the reproducibility was sometimes very low because of repetitive sequences around the primer sequences. With the present method, strong bands for the disease alleles were reproducibly visible in a conventional agarose gel stained with ethidium bromide without using isotopes. Three cases with sporadic HD and a case with senile chorea showed expanded alleles for HD with smaller sizes of the expansion than cases with typical HD. These results showed that the present method provides a simple and reproducible way to detect HD allele, and some cases with sporadic HD and senile chorea had expanded HD alleles.

Adult↗

Skin topography measurement by interference fringe projection: a technical validation.

BACKGROUND/AIMS: The quantitative analysis of skin topography is frequently used in cosmetology to evaluate the efficacy of hydrating or anti-wrinkle creams (micro-topography studies) or creams for slimming or to alleviate stretch marks (macro-topography studies). Numerous methods involving the three-dimensional (3-D) reconstruction of the topography from silicone replicas have been developed. Some of the main techniques applied include optical profilometry based on the reflection of oblique lighting or on transparency, and profilometry by laser focusing or triangulation. METHODS: We chose to test the Dermatop system developed by Eotech Co., which is based on interference fringe projection profilometry associated with the Toposurf surface processing software. Owing to the system's principle of operation, we first ran three experiments to evaluate the influence of ambient lighting on the 3-D reconstruction and the surface roughness parameters, the influence of the calibration device built into the acquisition system and the influence of calibration in terms of ambient lighting correction. We then evaluated the accuracy, the repeatability and the reproducibility of the measurements provided by the system from known metal topographic standards and skin replicas. RESULTS: It was seen that external lighting can cause variations in the calculation of parameters SPa, SPt and SPtm which can reach error levels of about 5% if the operator does not almost systematically calibrate the apparatus between each measurement. These three experiments enabled us to standardise the protocol for the acquisition of 3-D information, in order to minimise the lighting problems and yet respect the requirements for routine studies. The correlation coefficient between the theoretical values and the mean of experimental values was 0.9955 for parameter SPa and 0.9983 for parameter SPtm. Repeatability presented variations of under 4%, irrespective of the parameters, and reproducibility of under 6%. CONCLUSIONS: The technique provides very satisfactory results from the point of view of accuracy, repeatability and reproducibility. It is an excellent compromise between cost, accuracy and the time required.

Dermatology↗

Alfentanil: a study of its analgesic activity and interactions with morphine in the mouse.

Alfentanil has a short duration action which did not markedly increase with increasing dose in mice. It is approximately one-fourth as potent as fentanyl. Repeat injections gave reproducible analgesic effects but depression of respiratory rate increased when repeat injections were given on the basis of cessation of analgesic activity. Alfentanil appeared to reduce the analgesic effect of morphine given 30-60 min after alfentanil administration. This has been demonstrated with injection of alfentanil 15 min before, simultaneously with and 30 min after morphine injection. There were not significant effects on depression of respiratory rate produce by morphine.

Alfentanil↗

Polymorphous light eruption: successful reproduction of skin lesions, including papulovesicular light eruption, with ultraviolet B.

Thirty patients suffering from polymorphous light eruption (PLE), selected by criteria pointing to UVB sensitivity, were phototested for reproduction of skin lesions. Twenty-seven patients (90.0%) had symptoms compatible with or very similar to papulovesicular light eruption (PVLE). Of the 30 patients, 56.7% developed typical lesion of PLE at the test site by provocative phototesting with UVB. Eruptions of the immediate onset type were apt to be reproduced by multiple repeated irradiation, but those of the delayed onset type were reproduced by a single high-dose exposure. In addition, pruritus at the test site seen in all patients was thought to be an important diagnostic symptom. It was obvious that UVB played an etiological role in PLE, including PVLE.

Adolescent↗

The reproducibility and short-term and long-term repeatability of sonographic measurement of splenic length.

This study was undertaken to evaluate the reproducibility (interobserver variability), and the short-term and long-term repeatability (intraobserver variability) of sonographic measurement of the maximum and mean splenic length. Ultrasound (US) measurements of the splenic length were performed in 43 subjects, ages 30 to 59 years old (mean age = 44.5 years). Each subject attended three US sessions: baseline examination and 30 min and 8 weeks after the baseline examination. In each session, US examinations of the spleen were performed and the splenic length was measured by four operators. In each examination, the splenic length was measured 3 times, and the maximum and mean values were obtained. The overall reproducibility of the measurements of maximum and mean splenic lengths were 67% and 89%, respectively. The short-term repeatability in measuring the maximum and mean splenic lengths were 87% and 94%, respectively, whereas the long-term repeatabilities were 61% and 76%, respectively. The reproducibility and short-term and long-term repeatability of the measurement of mean splenic length were higher than those of the maximum splenic length. In the mean and maximum splenic length measurement, the short-term repeatability was higher than the long-term repeatability. The results suggested that the mean splenic length has a higher reliability in sonographic measurement and should be used in routine clinical practice. Measurement errors should be considered when evaluating the changes of splenic length in serial US examinations, particularly in long-term follow-up.

Adult↗

Validity and reproducibility of electrical impedance tomography for measurement of calf blood flow in healthy subjects.

The Sheffield electrical impedance tomography; (EIT) system produces images of changes in the distribution of resistivity within tissue. The paper reports on the application of electrical impedance tomography in monitoring volume changes in the limb during venous occlusion. The aim of the study is to assess the feasibility, reproducibility and validity of calf blood flow measurements by EIT. In 14 healthy volunteers calf blood flow is compared, as determined in a calf segment by strain-gauge plethysmography (SGP), with the impedance changes measured by EIT during rest and post-ischaemic hyperaemia. The measurements are repeated to assess reproducibility. The reproducibility for the EIT, assessed from the repeated measurements and expressed as a reproducibility coefficient, is 0.88 during rest and 0.89 during hyperaemia. The reproducibility coefficient for SGP data is 0.83 at rest and 0.67 during hyperaemia. Flow measurements, assessed by means of two methods, correlate well at rest (r = 0.89), but only moderately during hyperaemia (r = 0.51). The correlation coefficient for the pooled flow measurements is 0.98. It is concluded that EIT is a valid and reliable method for assessing blood flow in the limb. Possible applications of EIT in localising fluid changes are discussed.

Adult↗

Procedures for estimating confidence intervals for selected method performance parameters.

Procedures for estimating confidence intervals (CIs) for the repeatability variance (sigmar2), reproducibility variance (sigmaR2 = sigmaL2 + sigmar2), laboratory component (sigmaL2), and their corresponding standard deviations sigmar, sigmaR, and sigmaL, respectively, are presented. In addition, CIs for the ratio of the repeatability component to the reproducibility variance (sigmar2/sigmaR2) and the ratio of the laboratory component to the reproducibility variance (sigmaL2/sigmaR2) are also presented.

Analysis of Variance↗

Short and long term reproducibility of responsiveness to inhaled histamine: Rt compared to FEV1 as measurement of response to challenge.

Repeatability (123 min) and reproducibility (65 days) of response to inhaled histamine were tested in a selected group of stable asthmatics and non-asthmatics by means of FEV1 and breathing resistance (Rt) determined by the interruptor method. Variation in prechallenge FEV1 and Rt was kept within a +/- 10% and +/- 40% range, respectively. We found the PC20-FEV1 highly repeatable after 123 min. The 95% confidence interval for a single determination was the measured value +/- 1.72 two-fold concentration differences. Reproducibility was significantly lower (p less than 0.05). Both repeatability and reproducibility of PC30-Rt were significantly lower than that of PC20-FEV1. Interruptor method determined PC30-Rt was found repeatable within 4 two-fold concentration differences which is not acceptable for clinical and research purposes. The poor repeatability of PC30-Rt could be due to changes in functional residual capacity during challenge.

Adolescent↗

Reproducibility of the Conconi test: test repeatability and observer variations.

The repeatability of the the speed/heart rate (S/HR) relationship obtained with the incremental test developed by Conconi et al. (5) was examined by having 75 subjects perform the same running test twice in the course of a few days. From the data obtained, comparisons were made of four variables: 1) speed of deflection, 2) heart rate of deflection, 3) slope, and 4) intercept on the y-axis of the linear portion of the S/HR relationship. Straight-line equations, correlation coefficients (r), and technical errors of measurement (TEM) were obtained by comparing data from the two successive tests. Data analysis suggests that in two successive tests there are some variations in the linear portion of the S/HR relationship. However, the straight lines obtained from the two tests converge at the same deflection point. The problem of assigning a single value to the point of deflection, when determined by means of visual analysis of the S/HR graph, was also examined. For this part of the study, speed and heart rate data previously obtained for 65 subjects were used. The selected tests had been carried out during the course of various studies. The 65 sets of test data were plotted on graph paper and given to six observers for independent evaluation of speed and heart rate of deflection. These observers had different levels of experience in interpreting Conconi test results. The speed and heart rate of deflection values assigned by each observer were compared to the values obtained through mathematical analysis of the tests by computer. Straight-line equations, correlation coefficients (r), and technical errors of measurement (TEM) obtained by comparing visually determined data to those determined through mathematical analysis were calculated for each observer. Only for observers with little experience were some differences found between the observer-assigned and computer-determined results; these differences occurred for both speed and heart rate of deflection. We conclude that visual analysis provides information that is very similar to that obtained through computer analysis. The accuracy of the visually obtained information varies according to the observer's experience.

Adult↗

Measurement of radiographic joint space width in the tibiofemoral compartment of the osteoarthritic knee: comparison of standing anteroposterior and Lyon schuss views.

OBJECTIVE: To evaluate progression of joint space narrowing in radiographs of osteoarthritic (OA) knees imaged in both the standing anteroposterior (AP) and the Lyon schuss positions, using alternative methods to measure joint space width (JSW). METHODS: Standing AP (extended view) and Lyon schuss (posteroanterior [PA] view, with 20-30 degrees of flexion) radiographic images of 58 OA knees were obtained twice (at baseline and 2 years later). With both methods, fluoroscopy was used to align the anterior and posterior margins of the medial or lateral tibial plateau with the central x-ray beam. Minimum JSW, mean JSW, and joint space area (JSA) of the medial or lateral femorotibial joint space were measured using a new digital image analysis system. The effects of knee flexion versus extension and parallel versus nonparallel tibial plateau alignment were evaluated with respect to the reproducibility of JSW in repeated examinations (intraclass correlation coefficient [ICC]), the mean of within-knee standard deviations of repeated measurements (SD(m)), and the sensitivity to changes in JSW in serial radiographs (standardized response mean [SRM]). RESULTS: The performance of the new software, as assessed by the reproducibility of repeated measurements of minimum JSW on the same image, was excellent in both the standing AP (ICC = 0.98) and Lyon schuss radiographs (2 SD(m) = 0.5 mm, ICC = 0.98). The reproducibility in different radiographs of the same knee was not evaluated. However, over 2 years, the mean (+/- SD) decrease in the minimum JSW of OA knees was 0.17 +/- 0.75 mm in standing AP radiographs (P not significant) and 0.24 +/- 0.50 mm in Lyon schuss views (P = 0.007), with SRMs of 0.23 and 0.48, respectively. The quality of alignment of the tibial plateau was satisfactory (<1 mm between anterior and posterior margins of the medial tibial plateau) in 66% of the pairs of Lyon schuss radiographs and in 57% of the pairs of standing AP radiographs. In the Lyon schuss radiographs, SRM was highly dependent on tibial plateau alignment. Minimum JSW was more sensitive to change than was mean JSW or JSA, in paired Lyon schuss radiographs that exhibited satisfactory alignment. CONCLUSION: Compared with the standing AP radiograph, PA imaging of the knee in 20-30 degrees flexion (the schuss position) increases the reproducibility of radiographic JSW measurements in OA knees and the sensitivity to change in JSW in serial radiographs. Sensitivity to change in minimum JSW is notably increased by aligning the medial tibial plateau with the central x-ray beam in the Lyon schuss radiograph.

Aged↗

Mechanism of action of platelet activating factor in the pulmonary circulation: an investigation using a novel isotopic system in rabbit isolated lung.

1. Rabbit isolated lungs were perfused via the pulmonary artery with Tyrode solution containing 4.5% Ficoll and 0.1% bovine serum albumin at a constant rate of 20 ml min-1. Lung perfusate was drawn for alternating 5 min periods from two reservoirs, one containing 125I-albumin and the other unlabelled albumin to wash out the intravascular label. Microvascular 125I-albumin leakage was determined from the count remaining at the end of the washout phase with an external gamma scintillation probe. In addition, perfusion pressure was monitored continuously. Each experiment comprised 6 cycles over a total period of 60 min. 2. Infusion of platelet activating factor (PAF, 3 nmol min-1 for 10 min) resulted in microvascular 125I-albumin leakage, whereas lyso-PAF was without effect. During PAF infusions there was also an increase in perfusion pressure. Both the permeability and pressor effects of PAF were inhibited by the PAF antagonist L-652731. 3. Infusion of the thromboxane analogue U-46619 (0.6 nmol min-1 for 10 min) caused an increase in perfusion pressure but protein accumulation was not significantly different from that observed with control infusions. 4. Bolus injections of PAF (1 nmol) caused increases in perfusion pressure which were reduced by indomethacin, dazmegrel and BW 755C. Bolus injections of PAF, repeated at 30 min intervals caused reproducible pressor responses; however, repeated injections at 60 min intervals resulted in augmented responses. This augmentation did not occur in the presence of indomethacin. 5. Retrograde perfusion of PAF via the pulmonary vein induced increased perfusion pressure and microvascular 125I-albumin leakage. The observed increase in leakage when compared with forward perfusion suggests that PAF produces predominantly arteriolar constriction i.e. proximal to the site of leakage during forward perfusion. 6. These results indicate that PAF is a vasoconstrictor in the rabbit pulmonary circulation and augmented responses occur with repeated injections at 60 min intervals. Cyclo-oxygenase inhibition abolished this vascular hyperresponsiveness induced by PAF. PAF also caused protein accumulation in the lungs. Both these actions of PAF appear to be receptor-mediated because they were inhibited by PAF antagonists. Another pulmonary vasoconstrictor, U-46619 did not cause protein accumulation suggesting that the extravasation of protein with PAF is not merely secondary to changes in vascular tone.

15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗

Central corneal thickness measurement with the Pentacam Scheimpflug system, optical low-coherence reflectometry pachymeter, and ultrasound pachymetry.

PURPOSE: To assess the intraoperator repeatability and interoperator reproducibility of central corneal thickness measurements by the Pentacam Scheimpflug imaging system (Oculus) and the optical low-coherence reflectometer (OLCR) pachymeter (Haag-Streit) and to compare them with those of ultrasound (US) pachymetry. SETTING: Assaf Harofe Medical Center Ophthalmology Outpatient Clinic, Zerifin, Israel. METHODS: Repeatability was determined from 10 successive measurements in each of 4 healthy patients. Reproducibility for the Pentacam Scheimpflug system was determined from measurements by 2 operators in each of 24 patients; in these 24 patients, central corneal thickness measurements were compared between the Pentacam and US pachymetry. For the OLCR pachymeter, reproducibility was determined from measurements by 2 operators in each of 16 patients, in whom central corneal thickness was also measured with the Pentacam. RESULTS: Mean coefficient of repeatability was 0.84% for the Pentacam Scheimpflug system and 0.33% for the OLCR pachymeter. For the Pentacam, the coefficient of interoperator reproducibility was 1.10% and the 95% limits of agreement were -10.2 microm to +11.9 microm. Mean difference between Pentacam and US was 6.09 microm. For the OLCR pachymeter, the coefficient of interoperator reproducibility was 0.59% and the 95% limits of agreement were -5.4 microm to +7.0 microm. Mean difference between central corneal thickness values obtained with the OLCR pachymeter and Pentacam Scheimpflug system was 1.7 microm. CONCLUSIONS: Objective, noncontact measurement of central corneal thickness with the Pentacam Scheimpflug system and OLCR pachymeter was convenient and yielded excellent intraoperator repeatability and interoperator reproducibility. Central corneal thickness values obtained with the Pentacam were similar to those obtained with both the OLCR pachymeter and an US pachymeter. Further research is needed to corroborate whether central corneal thickness measurements by the Pentacam and OLCR devices can be used interchangeably and are more clinically useful than US pachymetry.

Biometry↗

Statistical analysis of a micromethod in an interlaboratory test.

A micromethod developed by Meier (1996) was assessed in an interlaboratory test with 7 labs supplying duplicate measurements on 5 to 9 days. This paper aims at evaluating the reproducibility and the repeatability of the micromethod. The effective concentration of microbicidal substances to reduce the number of test organisms by a factor of 10(5) was estimated from raw data of the microtiter test by means of a nonlinear regression model. Inclusion criteria were defined for the model fit and the results were checked for plausibility and for outliers according to methods prescribed by ISO 5725 (International Standard, 1994). From the remaining data (about 85% of the total data) inter-laboratory and intralaboratory reproducibility as well as repeatability were computed. The paper sets an example of how to analyze such interlaboratory tests. For practical purposes a table shows confidence limits of the relative precision for a broad range of interlaboratory test designs. In addition, test designs are given which lead to statistically qualified decisions about acceptance or rejection of a claimed effective concentration of a microbicidal substance. The results clearly qualify the micromethod for practical application in testing microbicidal substances.

Calibration↗

Reversibility and reproducibility of histamine induced plasma leakage in nasal airways.

Plasma exudation is one cardinal factor in airways defence and inflammation. In inflammatory airway diseases such as rhinitis and asthma, however, plasma leakage may also have a pathogenetic role. Experimental data from animals indicate that highly sensitive, active, and reversible processes regulate the vascular and mucosal permeability to macromolecules. With the use of a nasal lavage model for the recovery of liquids on the mucosal surface the effect of histamine on the macromolecular permeability of the airway endothelial-epithelial barriers was studied in normal subjects. The concentrations of albumin, kinins, and N-alpha-beta-tosyl-L-arginine-methyl esterase (TAME) in nasal lavage fluid were measured and nasal symptoms assessed by a scoring technique. The reproducibility of three repeated challenges with 30 minute intervals on the same day was studied in 12 subjects and compared with the same procedure (three challenges) on a different day. Sneezing decreased significantly (p less than 0.05) after the first histamine challenge but was maintained thereafter. Otherwise, the mean values for symptoms and for markers of vascular leakage were very similar both for the three challenges in the same session and for the two challenge sessions on a different day. Sneezing, blockage, and secretions were associated with increased concentrations of TAME esterase (maximum 9000 cpm/ml), kinins (1.4 ng/ml), and albumin (0.3 g/l) in lavage fluid. Both the symptoms and the measures of plasma exudation were reversible and reproducible in the three repeat histamine challenges and at two challenge sessions on different days. These findings support the view that non-injurious, active processes regulate the inflammatory flow of macromolecules across airways endothelial-epithelial barriers. The present experimental approach would be suitable for studies of the modulatory effects of inflammatory stimulus induced plasma leakage and symptoms in human airways.

Adult↗